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GOOD AFTERNOON,
EVERYONE.

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I'M NINA SCHOR, DEPUTY DIRECTOR

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FOR INTRAMURAL RESEARCH HERE AT

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NIH.

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IT IS MY PLEASURE AND MY HONOR

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TO INTRODUCE THE SYMPOSIUM.

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THIS IS DAY 4 OF OUR 2023

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RESEARCH FESTIVAL.

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AND THIS DAY ACTUALLY RECOGNIZES

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THE WORK OF OUR SENIOR

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INVESTIGATORS.

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BOY, I CAN TELL YOU, YOU ARE IN

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FOR A TREAT WITH THIS SYMPOSIUM

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COMING UP.

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THIS SYMPOSIUM WILL PRESENT AND

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SHOWCASE FIVE INVESTIGATORS FROM

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NIH WHO HAVE BEEN ELECTED IN THE

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LAST TWO YEARS TO THE NATIONAL

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ACADEMY OF THE SCIENCES.

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SO, FIRST A FEW WORDS ABOUT THE

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NATIONAL ACADEMIES.

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THE NATIONAL ACADEMIES PROVIDE

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RECOMMENDATIONS AND EXPERT

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ADVICE ON ISSUES OF IMPORTANCE

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TO SCIENCE, HEALTH, AND

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ENGINEERING.

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CURRENTLY, THERE ARE THREE

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NATIONAL ACADEMIES.

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NATIONAL ACADEMY OF SCIENCES,

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THE NATIONAL ACADEMY OF

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MEDICINE, AND THE NATIONAL

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ACADEMY OF ENGINEERING.

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AND NIH IS A PROLIFIC SOURCE OF

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ELECTEES TO THESE ACADEMIES, AND

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ELECTION TO ANY OF THESE

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ACADEMIES IS WIDELY DEEMED TO BE

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THE HIGHEST HONOR THAT THE

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SCIENTIFIC COMMUNITY CAN BESTOW

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UPON A SCIENTIST.

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WE CURRENTLY HAVE MORE THAN 50

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ACTIVE P.I.s IN THE NATIONAL

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ACADEMY OF SCIENCES, MORE THAN

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70 IN THE NATIONAL ACADEMY OF

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MEDICINE, AND ONE IN THE

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NATIONAL ACADEMY OF ENGINEERING.

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SO, REALLY A FERTILE GROUND FOR

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THE NURTURING OF THE SCIENCE AND

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THE P.I.s THAT UPON WHOM THIS

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HONOR IS BESTOWED.

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SO, OUR FIRST SPEAKER TODAY, I

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WILL INTRODUCE THE SPEAKERS

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INDIVIDUALLY AS THEY COME UP TO

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THE PODIUM OR ARE SEEN ON THE

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SCREEN, IS DR. JULIE SEGRE.

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AT THE OPENING OF THE FESTIVAL,

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I TALKED ABOUT A VIRUS THAT

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SHALL REMAIN UNNAMED, AND I'M

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STILL NOT GOING TO -- VOLDEMORT

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THE VIRUS, BUT IN ANY CASE, DR.

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SEGRE IS COMING TO US VIRTUALLY

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THIS AFTERNOON.

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SO SHE IS A SENIOR INVESTIGATOR

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AT NHGRI.

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SHE IS AN INTERNATIONALLY KNOWN

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EXPERT IN THE MICROBIOME AND

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PARTICULARLY THE MICROBIOME OF

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THE SKIN AND THE ROLES THAT

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PLAYS IN MEDIATING THE

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IMMUNOLOGIC EFFECTS OF THE SKIN

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AND THE PROTECTION THAT THE SKIN

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CONFERS UPON PEOPLE, AND THE

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ROLE OF THAT MICROBIOME IN 

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DISEASE.

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SHE APPLIES CUTTING EDGE GENOMIC

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AND GENE SEQUENCING TECHNOLOGIES

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TO BEST DEFINE THE MICROBIOME.

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AND SO WITHOUT FURTHER ADO, DR.

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SEGRE.

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>> THANK YOU VERY MUCH,
DR.

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SCHOR.

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IT IS A GREAT DISAPPOINTMENT TO

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ME NOT TO BE WITH YOU ALL, AND I

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HOPE I GET A RAIN CHECK SO THAT

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I CAN MEET YOU.

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BUT, YES, IT IS 3 1/2 YEARS

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LATER, AND YET HERE WE ARE.

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SO I WENT TO A -- THIS IS

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IRONIC, A MICROBIAL PATHOGENESIS

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MEETING, AND NOT SURPRISINGLY

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TWO TO THREE DAYS LATER I CAME

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HOME AND TESTED POSITIVE.

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SO, UNFORTUNATELY, I'M STILL IN

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MY PERIOD OF ISOLATION, BUT I'M

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GLAD I'M FEELING BETTER ENOUGH 

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TO JOIN YOU TODAY.

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REGRET NOT BEING WITH YOU ALL AT

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THE RESEARCH FESTIVAL.

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SO, I'M GOING TO KICK US OFF

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TODAY AND IT'S REALLY A PLEASURE

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TO TALK ABOUT MY WORK ON HUMAN

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MICROBIOME AND AS DR. SCHOR

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MENTIONED THE SKIN MICROBIOME IN

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PARTICULAR.

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AND THE TITLE IS FINDING OUR

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FRIENDS AMONGST THE FOES.

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AS MY WORK HAS BEEN ALSO AT THE 

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INTERSECTION OF ANAMICROBIAL

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RETANS AND MICROBIOME.

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PART OF MY WORK IS TO TALK WITH

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THE IDEA IT MIGHT INSPIRE ANY OF

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YOU AND IF YOU'RE INTERESTED IN

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DEVELOPING MICROBIOME AS PART OF

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YOUR OWN RESEARCH PROGRAM OR

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QUESTIONS THAT YOU HAVE, I WOULD

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WELCOME ANYONE TO SEND ME AN

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E-MAIL NOW OR IN THE FUTURE.

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THERE, OKAY.

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SO, LET'S JUST GET STARTED WITH

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THIS QUESTION OF WHAT DO I MEAN

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WHEN I SAY THE HUMAN MICROBIOME.

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BY THAT, I'M TALKING ABOUT THE

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COLLECTION OF ALL MICROORGANISMS

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AND THEIR DNA THAT LIVE IN

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ASSOCIATION WITH THE HUMAN BODY.

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SO I'M GOING TO GIVE A TALK

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TODAY AND COVER BOTH BACTERIA,

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FUNGI, AND VIRUSES.

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AND REALLY ONE OF THE GOALS OF

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THE HUMAN MICROBIOME PROJECT

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THAT I WAS PROUD TO BE A PART OF

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WAS TO MOVE THE DISCUSSION AWAY

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FROM THINKING ABOUT ONLY THE

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HUMAN MICROBIOME AND MICROBES

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THAT COLONIZE US AS BAD OR EVIL

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AND TO THINK AS MUCH ABOUT THE

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BENEFICIAL ROLES THAT THESE

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MICROBES PLAY ON OUR BODY.

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TO THINK ABOUT THIS AS TO

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REIMAGINE THAT WE THINK ABOUT

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OURSELVES, HUMANS, AS THE SUPER

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ORGANISMS, AND THAT WHEN I THINK

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ABOUT MYSELF, IT'S THAT I AM A

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COMBINATION OF THE HUMAN CELLS,

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SO THESE 30 TRILLION HUMAN

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CELLS, AS WELL AS AN EQUALLY

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LARGE NUMBER OF BACTERIA AND

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FUNGI AND PROBABLY EVEN MORE

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VIRUSES.

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BUT I CAME TO THIS WORK REALLY

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STARTING AS A GRADUATE

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OPPORTUNITY IN THE HUMAN GENOME

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PROJECT WHERE WE WERE WORKING TO

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DECIPHER THE HUMAN GENOME AND I

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THOUGHT THE ORIGINAL GOAL WAS WE

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WOULD STUDY THE HUMAN GENOME, 3

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BILLION BASE PAIRS OF DNA, AND

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ONCE REVEALED ABOUT THE 24,000

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PROTEIN CODING GENES THAT WE

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WOULD THEN HAVE THESE

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BREAKTHROUGHS, AND I THINK THOSE

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HAVE COME, EVEN MORE IS YET TO

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COME.

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WHEN WE THINK ABOUT HUMAN

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DISEASE, A LOT OF IT IS STILL 

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ABOUT THIS HOST-MICROBIAL

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INTERACTION AND AND MICROBES

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THAT STIMULATE IMMUNE SYSTEM,

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GENERATE RESPONSES OR CAUSE

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INFECTION.

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HERE IT'S A MORE COMPLEX GENETIC

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QUESTION.

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WHILE A BACTERIAL GENOME IS

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SMALL, SO LIKE LET'S SAY 3

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MILLION BASE PAIRS, RIGHT?

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SO A THOUSAND TIMES SMALLER THAN

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A HUMAN GENOME.

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AND THE PROTEIN CODING OF THAT

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IS TEN-FOLD LESS.

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BUT NOW WHEN YOU THINK ABOUT THE

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COMPLEXITY THAT THESE HUMAN

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CELLS ALL HAVE MORE OR LESS THE

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SAME CODING MATERIAL, BUT THE

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BACTERIA AND FUNGI CAN HAVE 

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DIFFERENT GENOMES ACROSS STRAINS

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AND SPECIES, YOU END UP WITH

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AGGREGATE WHERE THE COMPLEXITY

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OF THE GENOMES OF THE MICROBIOME

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IS EQUAL OR EVEN LARGER THAN THE

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HUMAN GENOME.

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THERE'S A VAST SPACE TO EXPLORE

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HERE.

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I WANT TO MAKE SURE WE START

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WITH THIS IDEA THAT THERE IS A

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TREMENDOUS BENEFIT THAT MICROBES

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PROVIDES, WHAT WE CALL SORT OF

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ECOSYSTEM SERVICES.

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SO, YOU KNOW, ONE OF THE GREAT

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THINGS THE MICROBES DO THAT I'VE

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HAD THE PLEASURE OF WORKING WITH

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MY COLLEAGUE, DR. YASMINE

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BELKADE, AN EXPERT, HOW MICROBES

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TUNE THE IMMUNE SYSTEM, MAKE

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SURE THE IMMUNE SYSTEM DOESN'T

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FALL ASLEEP, ALSO TO RECOGNIZE

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FRIENDS AND FOES.

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THE SKIN MICROBES ALSO ARE GOING

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TO SECRETE -- HELP BREAK DOWN

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PRODUCTS THAT THE HUMAN CELLS

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HAVE PRODUCED.

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AND HELP US THEN MAINTAIN THIS

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BARRIER.

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YOU SEE THIS IN THE GUT WHERE

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THEY WILL PROVIDE THIS BENEFIT

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OF HELPING US TO DIGEST FOOD AND

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NUTRITION.

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BUT REALLY ALSO ONE OF THE MAJOR

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THINGS THAT THE MICROBIOME DOES

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IS TO PROVIDE COLONIZATION

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RESISTANCE.

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IT'S MUCH MORE ENERGY INTENSIVE

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FOR SYSTEM TO MAINTAIN

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STERILITY.

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BUT YOU ARE CONSTANTLY BEING

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EXPOSED ON YOUR OUTER SURFACES

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TO PATHOGENS.

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IF YOU HAVE MICROBES ALREADY

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FILLING ALL THOSE NICHES, IT

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MAKES IT HARDER FOR PATHOGEN TO

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COME IN, AND TAKE UP RESIDENCE.

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SO, THIS IS ONE OF ALSO THE

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MAJOR BENEFITS OF HAVING

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MICROBES COLONIZE YOU.

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SO, MY GOAL AS SOMEONE IN THE

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BEGINNING OF THE MICROBIOME

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PROJECT HOW DO WE CHARACTERIZE A

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MICROBIOME, AND IN EARLY WORK

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THAT WE DID WITH DR. PATRICK

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MURRAY, WE THOUGHT ABOUT LIKE

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HOW ARE WE GOING TO CHARACTERIZE

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ALL THESE MICROBES?

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THE ORIGINAL WAYS HAD BEEN TO

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PUT THEM ONTO PETRI DISHES.

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AND THEN YOU WOULD GET THIS, YOU

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KNOW, THESE CULTURED ISOLATES,

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AND SOME OF THEM YOU WOULD SEE

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YOU COULD CULTURE BETTER ON, YOU

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KNOW, BLOOD AUGER OR BRAIN AND

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HEART INFUSION, SO YOU KNEW YOU

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WERE GETTING NOT THE FULL

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MICROBIAL COMMUNITY, BUT ABOUT

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HOW COMPLEX WITH MICROBIAL

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CULTURING.

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AT THIS POINT THE GENOME

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PROJECT, WHEN I STARTED GRADUATE

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SCHOOL IN 1990, WE'VE GONE

257
00:10:55,199 --> 00:11:00,137
THROUGH MAJOR REVOLUTION IN THE

258
00:11:00,137 --> 00:11:02,507
SCALE OF PROJECTS.

259
00:11:02,507 --> 00:11:03,241
ON 

260
00:11:03,241 --> 00:11:06,377
ON THE LEFT THE CLASSIC IMAGE OF

261
00:11:06,377 --> 00:11:08,145
READING THE GENOME IN BITS AND

262
00:11:08,145 --> 00:11:09,213
PIECES BUT THAT MEANT AT THE

263
00:11:09,213 --> 00:11:10,515
SAME TIME AS WE WERE

264
00:11:10,515 --> 00:11:11,916
UNDERSTANDING THE COMPLEXITY OF

265
00:11:11,916 --> 00:11:14,452
THE HUMAN GENOME, WE WERE HAVING

266
00:11:14,452 --> 00:11:16,320
A REVOLUTION IN THE SEQUENCING

267
00:11:16,320 --> 00:11:18,089
TECHNOLOGIES.

268
00:11:18,089 --> 00:11:20,358
AND MOVING FROM THE ORIGINAL

269
00:11:20,358 --> 00:11:23,728
SANGER SEQUENCING TO MUCH HIGHER

270
00:11:23,728 --> 00:11:24,762
DENSITY SEQUENCING PLATFORMS.

271
00:11:24,762 --> 00:11:27,231
ALONG WITH THAT CAME THE COMPUTE

272
00:11:27,231 --> 00:11:31,002
POWER THAT WE HAVE AT THE NIH

273
00:11:31,002 --> 00:11:35,006
THROUGH THE GREAT BENEFIT OF CIT

274
00:11:35,006 --> 00:11:38,476
AND BIOWULF TO ANALYZE THESE

275
00:11:38,476 --> 00:11:39,277
LARGE DATASETS.

276
00:11:39,277 --> 00:11:42,079
SO, I THINK OF SEQUENCING AS A

277
00:11:42,079 --> 00:11:44,448
NEW LENS TO OBSERVE BIOLOGY.

278
00:11:44,448 --> 00:11:49,620
AND WHAT WE -- THE SAME WAYS

279
00:11:49,620 --> 00:11:51,956
WE'RE CHARACTERIZING BACTERIA,

280
00:11:51,956 --> 00:11:54,292
THINKING OF GRAM POSITIVE OR

281
00:11:54,292 --> 00:11:55,593
GRAM NEGATIVE, UNDER MICROSCOPE

282
00:11:55,593 --> 00:11:57,728
OR STAINS WE USE, NOW WE'RE ALSO

283
00:11:57,728 --> 00:12:00,831
ADDING THIS OTHER FORM OF

284
00:12:00,831 --> 00:12:03,200
CHARACTERIZATION WHERE WE'RE

285
00:12:03,200 --> 00:12:07,371
SEQUENCING AND ABLE TO SEQUENCE

286
00:12:07,371 --> 00:12:11,075
MICROBIAL COMMUNITIES AS WELL AS

287
00:12:11,075 --> 00:12:12,410
INDIVIDUAL ISOLATES.

288
00:12:12,410 --> 00:12:13,477
WE'RE ILLUMINATING THIS MUCH

289
00:12:13,477 --> 00:12:15,479
GREATER SPACE BUT WE'RE THINKING

290
00:12:15,479 --> 00:12:16,414
ABOUT USING THAT INFORMATION IN

291
00:12:16,414 --> 00:12:19,584
THE SAME WAY SO I'M REALLY

292
00:12:19,584 --> 00:12:21,819
THINKING ABOUT, YOU KNOW, HOW

293
00:12:21,819 --> 00:12:25,723
THESE MICROBIAL COMMUNITIES COME

294
00:12:25,723 --> 00:12:26,557
TOGETHER.

295
00:12:26,557 --> 00:12:27,191
SORRY.

296
00:12:27,191 --> 00:12:30,528
THE FIRST STUDY WE DID ON THE

297
00:12:30,528 --> 00:12:32,196
SKIN MICROBIOME IN CONJUNCTION

298
00:12:32,196 --> 00:12:36,267
WITH MY LONG-TERM CLINICAL 

299
00:12:36,267 --> 00:12:38,102
COLLABORATOR DR. HEIDI KONG AND

300
00:12:38,102 --> 00:12:42,707
INSPIRED BY WORK WITH DR. MARIA

301
00:12:42,707 --> 00:12:46,110
TURNER, WAS TO REALLY PUT FORTH

302
00:12:46,110 --> 00:12:48,245
THIS IDEA THAT THE HUMAN BODY IS

303
00:12:48,245 --> 00:12:49,714
AN ECOSYSTEM.

304
00:12:49,714 --> 00:12:51,148
THIS WAS ONE OF OUR GREAT

305
00:12:51,148 --> 00:12:53,718
STRENGTHS AS YOU THINK ABOUT

306
00:12:53,718 --> 00:12:54,852
LIKE THE HUMAN MICROBIOME

307
00:12:54,852 --> 00:12:58,022
PROJECT A LOT HAS REALLY BEEN

308
00:12:58,022 --> 00:12:59,857
DRIVEN BY THE HUMAN GUT

309
00:12:59,857 --> 00:13:00,558
MICROBIOME.

310
00:13:00,558 --> 00:13:02,493
BUT THE SKIN HAS THIS REMARKABLE

311
00:13:02,493 --> 00:13:05,529
ABILITY THAT WE HAVE ACCESS TO

312
00:13:05,529 --> 00:13:06,097
THE TISSUE.

313
00:13:06,097 --> 00:13:08,432
MORE THAN EVEN JUST HAVING

314
00:13:08,432 --> 00:13:10,401
ACCESS, WE HAVE THESE

315
00:13:10,401 --> 00:13:12,570
CLINICAL -- THE CLINICAL ACUMEN

316
00:13:12,570 --> 00:13:14,839
OF DR. KONG, DR. TURNER, TO

317
00:13:14,839 --> 00:13:18,209
THINK ABOUT HOW DERMATOLOGIC

318
00:13:18,209 --> 00:13:21,212
DISORDERS MANIFEST AT SUCH

319
00:13:21,212 --> 00:13:22,079
STEREOTYPIC LOCATIONS, AND SO

320
00:13:22,079 --> 00:13:24,815
THE QUESTIONS THAT THEY BROUGHT

321
00:13:24,815 --> 00:13:26,717
FORTH WERE THIS IDEA ABOUT IS

322
00:13:26,717 --> 00:13:28,986
PSORIASIS ON THE OUTSIDE OF THE

323
00:13:28,986 --> 00:13:31,222
ELBOW AND ECZEMA ON THE INSIDE

324
00:13:31,222 --> 00:13:32,089
BECAUSE WE HAVE DIFFERENT

325
00:13:32,089 --> 00:13:33,958
MICROBIAL COMMUNITIES AS WELL AS

326
00:13:33,958 --> 00:13:35,026
HAVING DIFFERENT SKIN?

327
00:13:35,026 --> 00:13:41,098
SO THIS WAS A STUDY THAT DOCTORS

328
00:13:41,098 --> 00:13:42,566
KONG AND TURNER DESIGNED,

329
00:13:42,566 --> 00:13:44,669
LOOKING AT 20 BODY SITES, AND

330
00:13:44,669 --> 00:13:51,308
SURVEYED THEM BY LOOKING AT WHAT

331
00:13:51,308 --> 00:13:53,878
WERE THE BACTERIAL COMMUNITIES.

332
00:13:53,878 --> 00:13:56,280
THE OILY COMMUNITIES IN BLUE,

333
00:13:56,280 --> 00:13:57,314
BACTERIAL ARE SMALL PIECHARTS,

334
00:13:57,314 --> 00:14:06,824
YOU CAN SEE THE OILY SITES, LIKE

335
00:14:06,824 --> 00:14:12,229
FOREHEAD OR GLOBELLAR, THE NOSE,

336
00:14:12,229 --> 00:14:19,170
HAVE LIPID-LOVING BACTERIA,

337
00:14:19,170 --> 00:14:23,541
WHEREAS THE MOIST SITES WILL

338
00:14:23,541 --> 00:14:24,375
HAVE GREEN PROTEOBACTERIA,

339
00:14:24,375 --> 00:14:26,177
STAPHYLOCOCCUS, DRY SITES ARE

340
00:14:26,177 --> 00:14:27,611
SOME OF THE MOST DIVERSE

341
00:14:27,611 --> 00:14:28,279
BACTERIAL COMMUNITIES.

342
00:14:28,279 --> 00:14:31,048
AND IN FACT WHAT WE SAW WAS

343
00:14:31,048 --> 00:14:32,850
THAT, YOU KNOW, THE BEND OF MY

344
00:14:32,850 --> 00:14:33,851
RIGHT ELBOW IS MOST SIMILAR TO

345
00:14:33,851 --> 00:14:36,253
THE BEND OF MY LEFT ELBOW, BUT

346
00:14:36,253 --> 00:14:39,690
AFTER THAT, THE BEND OF MY LEFT

347
00:14:39,690 --> 00:14:41,125
ELBOW IS THE MOST SIMILAR TO THE

348
00:14:41,125 --> 00:14:44,795
BENDS OF YOUR LEFT ELBOW BECAUSE

349
00:14:44,795 --> 00:14:46,330
MICROBIAL COMMUNITIES ARE

350
00:14:46,330 --> 00:14:48,833
DEFINED BY THE ECOSYSTEM, BY THE

351
00:14:48,833 --> 00:14:51,368
NUTRIENTS AVAILABLE, RATHER THAN

352
00:14:51,368 --> 00:14:52,136
BY THE INDIVIDUAL.

353
00:14:52,136 --> 00:14:56,640
AND SO, YOU KNOW, MY AXILLA IS A

354
00:14:56,640 --> 00:14:57,742
SWEATY REGION SO THAT'S GOING TO

355
00:14:57,742 --> 00:15:01,445
BE QUITE DIFFERENT THAN A MOIST

356
00:15:01,445 --> 00:15:01,912
CREASE.

357
00:15:01,912 --> 00:15:03,748
THESE KINDS OF IDEAS ABOUT WHAT

358
00:15:03,748 --> 00:15:05,483
ARE THE COMMUNITIES WE DID THIS

359
00:15:05,483 --> 00:15:08,018
THEN WITH THE BACTERIA, WITH THE

360
00:15:08,018 --> 00:15:09,920
FUNGI, WITH THE VIRUSES, AND

361
00:15:09,920 --> 00:15:11,055
THESE KINDS OF CHARACTERIZATIONS

362
00:15:11,055 --> 00:15:13,924
THEN ENABLED US TO DO STUDY ON

363
00:15:13,924 --> 00:15:16,360
CLINICAL POPULATIONS SEEN HERE

364
00:15:16,360 --> 00:15:18,763
AT THE NIH, AND THIS WAS A

365
00:15:18,763 --> 00:15:22,566
COHORT OF CHILDREN WITH ATOPIC

366
00:15:22,566 --> 00:15:25,202
DERMATITIS, DR. KONG SEES.

367
00:15:25,202 --> 00:15:28,739
AND IN THIS CASE, CHILDREN WERE

368
00:15:28,739 --> 00:15:33,878
LOOKED AT, AT BASELINE, FLARE,

369
00:15:33,878 --> 00:15:35,646
EPISODIC EXACERBATIONS, AND PUS

370
00:15:35,646 --> 00:15:35,846
FLARE.

371
00:15:35,846 --> 00:15:37,815
DR. KONG'S QUESTION WAS ABOUT WE

372
00:15:37,815 --> 00:15:39,517
KNEW FROM EARLIER STUDY, THIS IS

373
00:15:39,517 --> 00:15:41,218
JUST MOST RECENT STUDY, WE KNEW

374
00:15:41,218 --> 00:15:43,120
FROM EARLIER STUDY AT THE FLARE

375
00:15:43,120 --> 00:15:53,631
SOME KIDS HAD HIGH LEVELS OF 

376
00:15:55,733 --> 00:16:01,705
STAPH  EPIDERMIS, STAPH AUREUS,

377
00:16:01,705 --> 00:16:04,074
WAS IT MULTIPLE OR ONE STRANGE

378
00:16:04,074 --> 00:16:05,876
AND HOW WERE THESE BACTERIAL

379
00:16:05,876 --> 00:16:09,180
STRAINS DRIVING THIS DISEASE.

380
00:16:09,180 --> 00:16:12,416
SO, IN THIS STUDY, SAMPLES WERE

381
00:16:12,416 --> 00:16:14,952
PUT FORTH FOR SEQUENCING, AND

382
00:16:14,952 --> 00:16:16,020
ALSO WE CULTURED INDIVIDUAL

383
00:16:16,020 --> 00:16:19,824
ISOLATES SO WE COULD DO

384
00:16:19,824 --> 00:16:20,724
STRAIN-LEVEL RESOLUTION.

385
00:16:20,724 --> 00:16:26,130
WHAT WE SAW IF THERE IS A CHILD 

386
00:16:26,130 --> 00:16:29,200
WHO HAS STAFF EPIDERMIDIS IN

387
00:16:29,200 --> 00:16:31,035
THEIR FLARE TIME POINT, SHOWING

388
00:16:31,035 --> 00:16:35,105
JUST THE MICROBIAL COMMUNITY

389
00:16:35,105 --> 00:16:38,108
THAT IS THE STAPHYLOCOCCUS, JUST

390
00:16:38,108 --> 00:16:39,910
A SINGLE STRAIN OF STAPH AUREUS,

391
00:16:39,910 --> 00:16:42,613
THAT WILL INCREASE IN THE FLARE,

392
00:16:42,613 --> 00:16:46,150
BUT THEN WE SEE IF YOU MOVE TO

393
00:16:46,150 --> 00:16:49,253
POST-FLARE TIME POINTS YOU'LL

394
00:16:49,253 --> 00:16:50,588
SEE INCREASE IN STAPH EPI, AN

395
00:16:50,588 --> 00:16:52,590
INTERESTING WAY THAT ALLOWS US

396
00:16:52,590 --> 00:16:57,061
TO BUILD A MODEL THAT MAYBE THE

397
00:16:57,061 --> 00:16:58,262
STAPH EPIIS HELPING RESOLVE THE

398
00:16:58,262 --> 00:16:59,230
FLARE OF THE STAPH AUREUS.

399
00:16:59,230 --> 00:17:01,599
IN THIS CASE ON THE RIGHT NOW

400
00:17:01,599 --> 00:17:04,168
HERE WHERE I HAVE A CHILD WHO

401
00:17:04,168 --> 00:17:07,771
HAS AT THE FLARE INCREASE IN A

402
00:17:07,771 --> 00:17:09,840
LOT OF STAPH EPIDERMIDIS AS WELL

403
00:17:09,840 --> 00:17:11,442
AS A LITTLE STAPH AUREUS, IN THE

404
00:17:11,442 --> 00:17:13,777
FLARE TIME POINT WE'RE SEEING

405
00:17:13,777 --> 00:17:18,148
INCREASE IN ALL OF THE STAPH

406
00:17:18,148 --> 00:17:22,219
EPIDERMIDIS SUGGESTING THERE'S

407
00:17:22,219 --> 00:17:24,855
SOMETHING ELSE IN TERMS OF

408
00:17:24,855 --> 00:17:26,357
INITIATION AND POST-FLARE STILL

409
00:17:26,357 --> 00:17:28,025
SEEING HIGH LEVELS OF STAPH

410
00:17:28,025 --> 00:17:28,592
EPIDERMIDIS.

411
00:17:28,592 --> 00:17:30,794
WE HAVE THESE STRAINS OF STAFF

412
00:17:30,794 --> 00:17:33,030
EPI AND STAPH AUREUS WE'RE ABLE

413
00:17:33,030 --> 00:17:34,798
TO USE IN ANIMAL MODELS AND

414
00:17:34,798 --> 00:17:36,800
SHARE WITH THE COMMUNITY TO

415
00:17:36,800 --> 00:17:42,172
START TO HAVE ANIMAL MODELS THAT

416
00:17:42,172 --> 00:17:43,774
REALLY THEN PUT FORTH THE

417
00:17:43,774 --> 00:17:46,510
LEGITIMATE SYSTEM OF HAVING THE

418
00:17:46,510 --> 00:17:49,146
HOST WITH THE, YOU KNOW,

419
00:17:49,146 --> 00:17:51,548
FILLEGRA MUTATION OR MOUSE MODEL

420
00:17:51,548 --> 00:17:53,183
WITH THE MUTATION, USING THESE

421
00:17:53,183 --> 00:17:54,718
STRAINS WE KNOW ARE INCREASING

422
00:17:54,718 --> 00:17:57,354
IN A FLARE AND SHOWING TO DRIVE

423
00:17:57,354 --> 00:17:57,855
DISEASE.

424
00:17:57,855 --> 00:18:04,328
WE HOPE BY DOING THIS WE'LL BE

425
00:18:04,328 --> 00:18:05,763
ABLE TO DEVELOP DIAGNOSTICS YOU

426
00:18:05,763 --> 00:18:07,398
CAN IMAGINE IN BASELINE STATE

427
00:18:07,398 --> 00:18:10,267
YOU CAN START TO MONITOR EITHER

428
00:18:10,267 --> 00:18:13,003
THE MICROBIAL COMMUNITY OR

429
00:18:13,003 --> 00:18:14,204
METABOLITES OF THE MICROBIAL

430
00:18:14,204 --> 00:18:16,407
COMMUNITY AND PREDICT WHEN A KID

431
00:18:16,407 --> 00:18:19,109
MIGHT TRANSITION FROM BASELINE

432
00:18:19,109 --> 00:18:20,444
TO FLARE, AND INTERVENE EARLIER

433
00:18:20,444 --> 00:18:25,582
THAN WHEN THE KID HAD ITCHED 

434
00:18:25,582 --> 00:18:28,218
THEMSELVES SO THEY ALREADY, YOU

435
00:18:28,218 --> 00:18:30,821
KNOW, EXCORIATED THE SKIN.

436
00:18:30,821 --> 00:18:32,823
 

437
00:18:32,823 --> 00:18:34,091
IN THE SKIN MICROBIOME WE HAVE

438
00:18:34,091 --> 00:18:37,494
THE GREAT PLEASURE OF BEING AT

439
00:18:37,494 --> 00:18:41,966
THE NIH CLINICAL CENTER, AND 

440
00:18:41,966 --> 00:18:44,234
WORKING ON THESE FASCINATING

441
00:18:44,234 --> 00:18:45,469
PATIENTS WITH INBORN ERRORS OF

442
00:18:45,469 --> 00:18:47,004
IMMUNITY SO WE CAN BEGIN TO

443
00:18:47,004 --> 00:18:50,441
UNDERSTAND WHAT IS THE

444
00:18:50,441 --> 00:18:50,975
HOST-MICROBE INTERACTION.

445
00:18:50,975 --> 00:18:55,746
WE THINK A LOT ABOUT HOW THE

446
00:18:55,746 --> 00:18:57,314
MICROBES STIMULATE THE IMMUNE

447
00:18:57,314 --> 00:18:59,516
SYSTEM, WE'RE ALSO FASCINATED TO

448
00:18:59,516 --> 00:19:02,786
COME UP WITH THE CONCEPT OF

449
00:19:02,786 --> 00:19:03,821
WHETHER THE MICROBES THEMSELVES,

450
00:19:03,821 --> 00:19:06,523
YOU KNOW, ARE BEING SEEN BY THE

451
00:19:06,523 --> 00:19:09,059
IMMUNE SYSTEM AND WHETHER THERE

452
00:19:09,059 --> 00:19:11,895
IS SPECIFICITY TO THAT.

453
00:19:11,895 --> 00:19:15,799
SO, THERE'S A LARGE COHORT OF

454
00:19:15,799 --> 00:19:18,602
GENETICALLY DEFINED PRIMARY OR

455
00:19:18,602 --> 00:19:20,337
IMMUNE -- PRIMARY IMMUNE

456
00:19:20,337 --> 00:19:22,506
DEFICIENT PATIENTS,

457
00:19:22,506 --> 00:19:24,241
FASCINATINGLY THEY HAVE -- YOU

458
00:19:24,241 --> 00:19:27,211
KNOW, MANY PRESENT WITH ATOPIC

459
00:19:27,211 --> 00:19:27,878
DERMATITIS WHICH GOT US

460
00:19:27,878 --> 00:19:33,350
ORIGINALLY INTERESTED BUT WILL

461
00:19:33,350 --> 00:19:36,754
ALSO HAVE OTHER ATOPES, AND WE

462
00:19:36,754 --> 00:19:40,691
LOOKED ON THE LEFT AT DOCK8

463
00:19:40,691 --> 00:19:42,659
DEFICIENT PATIENTS WITH ATOPY

464
00:19:42,659 --> 00:19:45,396
BUT A LOT OF WARTS, AND GOT THE

465
00:19:45,396 --> 00:19:50,868
FEELING THERE MIGHT BE

466
00:19:50,868 --> 00:19:52,269
ADDITIONAL HPVs.

467
00:19:52,269 --> 00:19:55,906
WHEN WE MINED GENOMIC DATA FROM

468
00:19:55,906 --> 00:20:00,144
THE DOCK8 PATIENTS WE WERE ABLE

469
00:20:00,144 --> 00:20:03,147
TO DEFINE 115 NEW HPV VARIANTS

470
00:20:03,147 --> 00:20:06,617
INCLUDING ONE NO SPECIES, THESE

471
00:20:06,617 --> 00:20:08,452
ARE THE 27 DOCK8 PATIENTS AND

472
00:20:08,452 --> 00:20:11,855
YOU CAN SEE WHERE I LABELED THE

473
00:20:11,855 --> 00:20:13,524
NOVEL SPECIES IN THE LIGHT BLUE

474
00:20:13,524 --> 00:20:17,561
AND LIGHT GREEN YOU CAN SEE

475
00:20:17,561 --> 00:20:20,130
THERE'S OFTEN MORE NOVEL SPECIES

476
00:20:20,130 --> 00:20:21,498
BEING DETECTED THAN KNOWN

477
00:20:21,498 --> 00:20:24,868
SPECIES OF HPV. 

478
00:20:24,868 --> 00:20:27,304
WE'VE EXPANDED THE REPERTOIRE.

479
00:20:27,304 --> 00:20:33,043
AS WELL WORKING WITH DR.

480
00:20:33,043 --> 00:20:33,744
NOTORANGELO HAVE LOOKED AT HIS

481
00:20:33,744 --> 00:20:35,479
RAG DEFICIENT PATIENTS.

482
00:20:35,479 --> 00:20:39,016
HERE AGAIN IT'S THIS INTERESTING

483
00:20:39,016 --> 00:20:41,318
IDEA OF WHETHER THERE IS

484
00:20:41,318 --> 00:20:44,054
INCREASED COLONIZATION IN

485
00:20:44,054 --> 00:20:44,955
PATIENTS WITH IMMUNE

486
00:20:44,955 --> 00:20:45,289
DEFICIENCIES.

487
00:20:45,289 --> 00:20:47,858
WE SAW THIS IN THE DOCK8

488
00:20:47,858 --> 00:20:49,793
PATIENTS.

489
00:20:49,793 --> 00:20:53,363
WE SAW IN THE NARES THEY CARRIED

490
00:20:53,363 --> 00:20:57,634
CORONAVIRUS, WE SAW THAT AGAIN

491
00:20:57,634 --> 00:21:00,237
IN THE RAG-DEFICIENT PATIENTS

492
00:21:00,237 --> 00:21:02,306
THAT WE IDENTIFIED RAG-DEFICIENT

493
00:21:02,306 --> 00:21:03,607
PATIENT WITH NOVEL CORONAVIRUS

494
00:21:03,607 --> 00:21:08,245
229E AS WELL AS SEEING

495
00:21:08,245 --> 00:21:08,912
ADDITIONAL NOROVIRUS, INFLUENZA,

496
00:21:08,912 --> 00:21:11,048
RHINOVIRUSES IN BOTH OF THESE

497
00:21:11,048 --> 00:21:11,348
POPULATIONS.

498
00:21:11,348 --> 00:21:12,082
WHAT'S REALLY INTERESTING ABOUT

499
00:21:12,082 --> 00:21:15,953
THAT OF COURSE IS THAT WE'D LIKE

500
00:21:15,953 --> 00:21:16,987
FOR PANDEMIC PREPAREDNESS HAVE A

501
00:21:16,987 --> 00:21:18,956
BETTER SENSE OF WHAT ARE THE

502
00:21:18,956 --> 00:21:21,391
VIRUSES THAT CAN COLONIZE HUMANS

503
00:21:21,391 --> 00:21:24,428
AND I SEE THE NIH CLINICAL

504
00:21:24,428 --> 00:21:25,929
CENTER'S PATIENTS WITH PRIMARY

505
00:21:25,929 --> 00:21:28,031
IMMUNE DEFICIENCY AS A RICH

506
00:21:28,031 --> 00:21:29,333
RESOURCE FOR UNDERSTANDING

507
00:21:29,333 --> 00:21:31,301
VIRUSES THAT CAN COLONIZE HUMANS

508
00:21:31,301 --> 00:21:33,170
AS WELL AS USING THIS

509
00:21:33,170 --> 00:21:34,905
INFORMATION TO HELP TAILOR THEIR

510
00:21:34,905 --> 00:21:37,241
CARE BECAUSE WE'RE OFTEN FINDING

511
00:21:37,241 --> 00:21:39,309
VIRUSES THAT PERHAPS WE DIDN'T

512
00:21:39,309 --> 00:21:43,780
EVEN KNOW TO TEST FOR.

513
00:21:43,780 --> 00:21:45,115
ENOUGH ABOUT THE MICROBIUM

514
00:21:45,115 --> 00:21:46,016
FRIENDS AND, YOU KNOW, LET ME

515
00:21:46,016 --> 00:21:47,985
TALK ALSO A LITTLE ABOUT THE

516
00:21:47,985 --> 00:21:50,554
FOES BECAUSE THIS HAS BEEN SOME

517
00:21:50,554 --> 00:21:54,491
OF THE WORK THAT REALLY HAS BEEN

518
00:21:54,491 --> 00:21:58,595
FOR ME PERFORMANCE FOREFRONT OF

519
00:21:58,595 --> 00:22:02,499
HOW RESEARCH CAN BE EMBEDDED

520
00:22:02,499 --> 00:22:04,535
INTO HOSPITAL LIKE THE NIH

521
00:22:04,535 --> 00:22:07,838
CLINICAL CENTER, SO HERE IN THIS

522
00:22:07,838 --> 00:22:08,505
TALK ABOUT THE NATIONAL

523
00:22:08,505 --> 00:22:10,908
ACADEMIES, I WANT TO BRING BACK

524
00:22:10,908 --> 00:22:13,076
TO A DECADE AGO, WHICH WAS

525
00:22:13,076 --> 00:22:15,746
REALLY ONE OF THE SIGNATURES OF

526
00:22:15,746 --> 00:22:18,482
MY CAREER AT NIH WHICH WAS THE

527
00:22:18,482 --> 00:22:24,955
OPPORTUNITY TO WORK WITH DR. 

528
00:22:24,955 --> 00:22:26,823
TARA PALMORE, DAVID HENDERSON.

529
00:22:26,823 --> 00:22:29,293
AND IN THIS CASE, IT WAS THE

530
00:22:29,293 --> 00:22:30,961
FIRST PATIENT WHO WAS RECOGNIZED

531
00:22:30,961 --> 00:22:36,200
AS COMING TO THE NIH CLINICAL

532
00:22:36,200 --> 00:22:41,772
CENTER CARRYING A CARBAPENEM

533
00:22:41,772 --> 00:22:46,076
RESISTANT KLEBSIELLA PNEUMONIA.

534
00:22:46,076 --> 00:22:55,786
THE ENTEROBACTER ESIA LEAD THE

535
00:22:55,786 --> 00:22:57,120
THREATS, NO ANTIBIOTICS LEFT TO

536
00:22:57,120 --> 00:22:58,488
TREAT THESE BACTERIA.

537
00:22:58,488 --> 00:23:01,058
IN THIS STORY WE HAD A PATIENT

538
00:23:01,058 --> 00:23:05,195
COME INTO THE CLINICAL CENTER,

539
00:23:05,195 --> 00:23:06,563
AND SHE WAS PUT IN ISOLATION AT

540
00:23:06,563 --> 00:23:07,764
THAT POINT.

541
00:23:07,764 --> 00:23:12,736
WE HAD A FAIR AMOUNT OF

542
00:23:12,736 --> 00:23:13,904
EXPERIENCE AND PUT HER IN THE

543
00:23:13,904 --> 00:23:18,342
LAST ROOM OF THE ICU.

544
00:23:18,342 --> 00:23:20,944
AND A GOOD TWO MONTHS WENT BY,

545
00:23:20,944 --> 00:23:24,681
AND THEN THERE WAS A SECOND,

546
00:23:24,681 --> 00:23:25,549
THIRD -- OOPS.

547
00:23:25,549 --> 00:23:26,450
SORRY.

548
00:23:26,450 --> 00:23:28,719
SECOND, THIRD, FOURTH AND FIFTH

549
00:23:28,719 --> 00:23:34,725
PATIENT WHO WERE IDENTIFIED AS

550
00:23:34,725 --> 00:23:36,293
CARRYING KPC, KRE KPC.

551
00:23:36,293 --> 00:23:38,228
THE TIMELINE IS WHEN PATIENTS

552
00:23:38,228 --> 00:23:40,197
WERE IDENTIFIED AND THE ORANGE

553
00:23:40,197 --> 00:23:42,566
ONES WERE IDENTIFIED AS CLINICAL

554
00:23:42,566 --> 00:23:43,700
CULTURES, BLUE IDENTIFIED AS

555
00:23:43,700 --> 00:23:45,102
SURVEILLANCE CULTURES.

556
00:23:45,102 --> 00:23:50,607
THESE WERE VERY SICK PATIENTS IN

557
00:23:50,607 --> 00:23:51,408
THE ICU.

558
00:23:51,408 --> 00:23:53,977
AND SO I CALLED UP DR. HENDERSON

559
00:23:53,977 --> 00:23:55,412
AND SAID, I'D LIKE TO SEQUENCE

560
00:23:55,412 --> 00:23:58,048
THEM AND LET'S SEE IF WE CAN

561
00:23:58,048 --> 00:23:59,549
FIGURE OUT ANYTHING.

562
00:23:59,549 --> 00:24:00,784
WE TOOK PATIENT NUMBER 1 BECAUSE

563
00:24:00,784 --> 00:24:03,053
WE THOUGHT THAT WAS GOING TO BE

564
00:24:03,053 --> 00:24:03,987
THE OUTLIER, AND THOUGHT THAT'S

565
00:24:03,987 --> 00:24:06,456
HOW WE WERE GOING TO UNDERSTAND

566
00:24:06,456 --> 00:24:08,425
WHAT'S THE SIMILARITIES BETWEEN

567
00:24:08,425 --> 00:24:17,134
2, 3, 4 AND 5 USING PATIENT 1 AS

568
00:24:17,134 --> 00:24:17,467
THE REFERENCE.

569
00:24:17,467 --> 00:24:20,137
THIS IS THE DECISION POINT.

570
00:24:20,137 --> 00:24:22,239
OUR PRIMARY QUESTION WAS ABOUT

571
00:24:22,239 --> 00:24:23,540
THE RELATEDNESS OF PATIENTS 2,

572
00:24:23,540 --> 00:24:29,246
3, 4 AND 5, BUT WE ALSO HAD THIS

573
00:24:29,246 --> 00:24:30,647
REALLY DEEP QUESTION ABOUT

574
00:24:30,647 --> 00:24:34,151
PATIENT 1 AND WAS PATIENT 1

575
00:24:34,151 --> 00:24:37,888
GOING TO MATCH 2, 3, 4 AND 5.

576
00:24:37,888 --> 00:24:41,391
 THE REASON THAT WAS IMPORTANT,

577
00:24:41,391 --> 00:24:42,492
THE DECISION MATRIX DR. PALMORE

578
00:24:42,492 --> 00:24:44,895
HAS TO MAKE, WHICH IS THE

579
00:24:44,895 --> 00:24:45,962
QUESTION IF PATIENT 1 MATCHES 2,

580
00:24:45,962 --> 00:24:51,568
3, AND 4 AND WE'RE ON THE LEFT,

581
00:24:51,568 --> 00:24:53,370
THEN WE HAD NOSOCOMIAL, OR

582
00:24:53,370 --> 00:24:54,037
HOSPITAL TRANSMISSION.

583
00:24:54,037 --> 00:24:57,140
THAT WOULD MEAN THAT THE CONTACT

584
00:24:57,140 --> 00:24:58,208
ISOLATION WAS INSUFFICIENT, THE

585
00:24:58,208 --> 00:25:00,610
SCREENING OF PATIENTS WAS

586
00:25:00,610 --> 00:25:02,145
INSUFFICIENT, AND THERE WAS A

587
00:25:02,145 --> 00:25:04,848
NEED THEREFORE TO CHANGE THE

588
00:25:04,848 --> 00:25:06,383
HOSPITAL PRACTICE.

589
00:25:06,383 --> 00:25:08,752
ON THE OTHER HAND, IF PATIENT 1

590
00:25:08,752 --> 00:25:11,788
DOESN'T MATCH 2, 3, 4 AND 5,

591
00:25:11,788 --> 00:25:12,823
IT'S TWO SEPARATE INTRODUCTIONS,

592
00:25:12,823 --> 00:25:15,359
AND THEN THAT MEANS THE CONTACT

593
00:25:15,359 --> 00:25:16,626
ISOLATION IS SUFFICIENT,

594
00:25:16,626 --> 00:25:18,428
SCREENING OF PATIENTS IS

595
00:25:18,428 --> 00:25:20,864
SUFFICIENT, AND STAY THE COURSE.

596
00:25:20,864 --> 00:25:22,099
THAT'S MORE ACTUALLY ON THE

597
00:25:22,099 --> 00:25:22,966
RIGHT-HAND SIDE, THAT'S MORE

598
00:25:22,966 --> 00:25:24,968
WHAT HAD HAPPENED TO THE NIH

599
00:25:24,968 --> 00:25:32,342
CLINICAL CENTER DURING

600
00:25:32,342 --> 00:25:32,676
ASENITOBACTER.

601
00:25:32,676 --> 00:25:35,445
WE DIDN'T HAVE THE RESOLUTION,

602
00:25:35,445 --> 00:25:37,280
MICRO LAB STILL RUNNING GELS,

603
00:25:37,280 --> 00:25:39,282
ALL WOULD LOOK THE SAME.

604
00:25:39,282 --> 00:25:44,187
BECAUSE 70% OF HEALTH CARE

605
00:25:44,187 --> 00:25:44,788
ACQUIRED KLEBSIELLA PMEUMONIAE

606
00:25:44,788 --> 00:25:46,957
IN THE U.S. ARE SEQUENCE TYPE

607
00:25:46,957 --> 00:25:48,558
258 THAT LOOK THE SAME.

608
00:25:48,558 --> 00:25:55,265
COULD GENOMIC DATA GIVE US MORE

609
00:25:55,265 --> 00:26:00,036
INFORMATION TO RESOLVE THIS?

610
00:26:00,036 --> 00:26:00,237
OKAY.

611
00:26:00,237 --> 00:26:01,838
SO, PATIENT 1, I WANT TO TAKE A

612
00:26:01,838 --> 00:26:04,674
BIT OF A DEEPER DIVE.

613
00:26:04,674 --> 00:26:06,476
NOW, ON THE LEFT-HAND SIDE I'VE

614
00:26:06,476 --> 00:26:08,512
DRAWN THESE DIAGRAMS.

615
00:26:08,512 --> 00:26:12,382
THIS IS PATIENT NUMBER 1, AND

616
00:26:12,382 --> 00:26:14,418
SHE HAD BEEN COLONIZED FOR SO

617
00:26:14,418 --> 00:26:16,753
LONG THAT SHE ACTUALLY WAS

618
00:26:16,753 --> 00:26:19,656
COLONIZED AT MULTIPLE SITES IN

619
00:26:19,656 --> 00:26:20,690
HER BODY.

620
00:26:20,690 --> 00:26:22,259
WE HAD ISOLATES, MULTIPLE

621
00:26:22,259 --> 00:26:23,093
ISOLATES, FROM URINARY TRACT

622
00:26:23,093 --> 00:26:26,029
OVER A WEEK OR TWO IN WHICH WE

623
00:26:26,029 --> 00:26:28,031
KEPT CULTURES AND CLINICAL

624
00:26:28,031 --> 00:26:29,966
MICROLAB KEPT CULTURING THEM.

625
00:26:29,966 --> 00:26:31,868
WE ALSO HAD ISOLATES FROM THE

626
00:26:31,868 --> 00:26:35,138
THROAT AND FROM THE GROIN AREA.

627
00:26:35,138 --> 00:26:37,841
SO WE SEQUENCED ALL OF THESE

628
00:26:37,841 --> 00:26:38,108
ISOLATES.

629
00:26:38,108 --> 00:26:42,412
AND WHAT I'M SHOWING YOU IS THAT

630
00:26:42,412 --> 00:26:44,114
ON THE VERY LEFT, THE PLAIN

631
00:26:44,114 --> 00:26:45,182
CIRCLE, THAT'S THE URINE

632
00:26:45,182 --> 00:26:46,082
ISOLATE.

633
00:26:46,082 --> 00:26:48,418
WE'RE GOING TO CONSIDER THAT

634
00:26:48,418 --> 00:26:50,086
ANCESTRAL OR REFERENCE STRAIN.

635
00:26:50,086 --> 00:26:52,589
IN THE THROAT ISOLATE, THERE

636
00:26:52,589 --> 00:26:55,659
SEEMS TO HAVE BEEN THREE SINGLE

637
00:26:55,659 --> 00:26:58,428
NUCLEOSIDE CHANGES IN A 6

638
00:26:58,428 --> 00:26:59,062
MEGABASE GENOME.

639
00:26:59,062 --> 00:27:00,397
THAT WOULD HAVE HAPPENED NOT IN

640
00:27:00,397 --> 00:27:02,232
CODING GENE OR ANYTHING BUT

641
00:27:02,232 --> 00:27:03,800
SMALL ERRORS IN REPLICATION THAT

642
00:27:03,800 --> 00:27:07,370
GET PASSED ON, IN A CLONAL

643
00:27:07,370 --> 00:27:08,071
SPREAD.

644
00:27:08,071 --> 00:27:11,208
THE GROIN SEEMS TO HAVE BEEN

645
00:27:11,208 --> 00:27:13,477
SEPARATE, YOU KNOW, SEPARATE

646
00:27:13,477 --> 00:27:16,246
COLONIZATION, YOU SEE THREE 

647
00:27:16,246 --> 00:27:17,781
DIFFERENT SNPs.

648
00:27:17,781 --> 00:27:21,151
THE KEY, THESE SNPs ARE

649
00:27:21,151 --> 00:27:22,152
IDENTIFIABLE TO THIS PATIENT.

650
00:27:22,152 --> 00:27:25,255
SO THE MOMENT THAT WE ALL

651
00:27:25,255 --> 00:27:27,757
REMEMBER IS SITTING THERE WITH

652
00:27:27,757 --> 00:27:29,192
THE ORIGINAL SEQUENCE DATA, WE

653
00:27:29,192 --> 00:27:32,462
GOT THE ISOLATES AND HAD THE

654
00:27:32,462 --> 00:27:33,864
SEQUENCE DATA, AND WITHIN 24

655
00:27:33,864 --> 00:27:35,532
HOURS WE HAD ANALYZED IT AND

656
00:27:35,532 --> 00:27:37,267
WE'RE SITTING IN A CONFERENCE

657
00:27:37,267 --> 00:27:40,437
ROOM WITH DR. PALMORE AND SAY,

658
00:27:40,437 --> 00:27:43,673
WE HAVE A MATCH FROM PATIENT 1,

659
00:27:43,673 --> 00:27:44,941
AND THE OTHER PATIENTS.

660
00:27:44,941 --> 00:27:47,978
BUT THE PART WE DON'T UNDERSTAND

661
00:27:47,978 --> 00:27:52,482
IS THAT WE THINK PATIENT 1

662
00:27:52,482 --> 00:27:53,583
TRANSMITTED TO PATIENT 3,

663
00:27:53,583 --> 00:27:55,986
BECAUSE THAT'S WHERE YOU SEE THE

664
00:27:55,986 --> 00:27:57,854
SAME THREE SNPs IN PATIENT 3'S

665
00:27:57,854 --> 00:28:00,190
THROAT ISOLATE AS IN PATIENT 1.

666
00:28:00,190 --> 00:28:04,027
AND THEN WE BELIEVE PATIENT 3

667
00:28:04,027 --> 00:28:05,395
TRANSMITTED TO PATIENT 2.

668
00:28:05,395 --> 00:28:06,796
BECAUSE PATIENT 2 HAS AN

669
00:28:06,796 --> 00:28:10,333
ADDITIONAL SNP WE DIDN'T SEE IN 

670
00:28:10,333 --> 00:28:12,969
EITHER PATIENT 3 OR 1.

671
00:28:12,969 --> 00:28:14,471
AND DR. PALMORE SAYS, YEAH,

672
00:28:14,471 --> 00:28:16,773
THAT'S ALSO WHAT I THINK

673
00:28:16,773 --> 00:28:17,107
HAPPENED.

674
00:28:17,107 --> 00:28:19,509
SO FOR THIS MOMENT WE HAD THE

675
00:28:19,509 --> 00:28:20,744
GENOMIC DATA AND EPIDEMIOLOGIC

676
00:28:20,744 --> 00:28:22,412
DATA LINING UP.

677
00:28:22,412 --> 00:28:25,348
THE BOTTOM I SHOW PATIENT TRACE

678
00:28:25,348 --> 00:28:26,249
DATA.

679
00:28:26,249 --> 00:28:29,686
PATIENT 1 AND PATIENT 3 IN

680
00:28:29,686 --> 00:28:32,455
TURQUOISE OVERLAPPED IN THE ICU.

681
00:28:32,455 --> 00:28:34,624
AFTER PATIENT 1 HAD ALREADY LEFT

682
00:28:34,624 --> 00:28:37,928
THE ICU, PATIENT 3 AND 2

683
00:28:37,928 --> 00:28:39,062
OVERLAPPED IN THE ICU.

684
00:28:39,062 --> 00:28:40,664
WHAT THIS WOULD SUGGEST IS THAT

685
00:28:40,664 --> 00:28:43,633
THERE WAS A TRANSMISSION FROM

686
00:28:43,633 --> 00:28:46,403
PATIENT 1 TO 3, THAT WENT

687
00:28:46,403 --> 00:28:47,771
UNDETECTED FOR TWO MONTHS.

688
00:28:47,771 --> 00:28:50,974
BUT WAS CARRIED BY THAT PATIENT

689
00:28:50,974 --> 00:28:52,976
ENOUGH THAT THE PATIENT 3 COULD

690
00:28:52,976 --> 00:28:57,080
GO ON AND TRANSMIT TO PATIENT 2.

691
00:28:57,080 --> 00:28:58,848
I REALLY DO THINK PROBABLY THE

692
00:28:58,848 --> 00:29:01,017
WORLD OF COVID HAS TAUGHT US

693
00:29:01,017 --> 00:29:02,218
LIKE HOW THIS COULD HAPPEN BUT

694
00:29:02,218 --> 00:29:03,320
I'M TELLING YOU THIS WAS MIND

695
00:29:03,320 --> 00:29:06,389
BLOWING TO US AT THE TIME.

696
00:29:06,389 --> 00:29:08,191
AND IT MEANT EXACTLY THAT.

697
00:29:08,191 --> 00:29:13,630
WE HAD TO CHANGE THE -- ENHANCE 

698
00:29:13,630 --> 00:29:15,198
SURVEILLANCE OF ASYMPTOMATIC

699
00:29:15,198 --> 00:29:18,335
PATIENTS, AND HAVE MORE CONTACT

700
00:29:18,335 --> 00:29:18,602
ISOLATION.

701
00:29:18,602 --> 00:29:21,237
THERE CONTINUED TO BE A CLUSTER

702
00:29:21,237 --> 00:29:24,074
OF INFECTIONS AT THE NIH

703
00:29:24,074 --> 00:29:29,446
CLINICAL CENTER, BUT WITH THIS

704
00:29:29,446 --> 00:29:34,584
ENHANCED PRECAUTIONS THERE WAS A

705
00:29:34,584 --> 00:29:37,420
HALT IN THE TRANSMISSIONS THAT

706
00:29:37,420 --> 00:29:38,521
OCCURRED IN THE HOSPITAL.

707
00:29:38,521 --> 00:29:40,256
WE WERE ABLE TO UNDERSTAND WHAT

708
00:29:40,256 --> 00:29:42,058
WERE THE WEAKNESSES AND HOW WE

709
00:29:42,058 --> 00:29:44,828
COULD CHANGE THE PRACTICE.

710
00:29:44,828 --> 00:29:46,496
SO, A LOT OF WHAT WE DID WITH

711
00:29:46,496 --> 00:29:48,031
GENOMIC DATA WAS THINK ABOUT THE

712
00:29:48,031 --> 00:29:49,332
CONNECTIONS THAT WE COULD MAKE,

713
00:29:49,332 --> 00:29:52,602
BUT WE ALSO WERE ABLE TO RULE

714
00:29:52,602 --> 00:29:53,136
OUT CONNECTIONS.

715
00:29:53,136 --> 00:29:55,438
SO ON THE LEFT OF THIS SLIDE,

716
00:29:55,438 --> 00:29:57,540
I'VE DRAWN SORT OF WHAT WOULD

717
00:29:57,540 --> 00:29:59,976
HAVE BEEN THE POSSIBLE

718
00:29:59,976 --> 00:30:01,277
CONNECTIONS WHERE IF YOU HAD

719
00:30:01,277 --> 00:30:03,146
LOOKED AT HOW ALL THE WAYS IN

720
00:30:03,146 --> 00:30:05,649
WHICH THESE PATIENTS WERE ON THE

721
00:30:05,649 --> 00:30:08,918
SAME BOARD OR ARE THE SAME

722
00:30:08,918 --> 00:30:09,452
MEDICAL DEEM WHAT WERE

723
00:30:09,452 --> 00:30:11,621
EPIDEMIOLOGIC LINKS BUT WHEN YOU

724
00:30:11,621 --> 00:30:13,223
COMBINE EPIDEMIOLOGIC LINKS WITH

725
00:30:13,223 --> 00:30:14,424
THE GENOMIC DATA, THEN YOU CAN

726
00:30:14,424 --> 00:30:17,494
SEE WHAT ARE THE MOST LIKELY

727
00:30:17,494 --> 00:30:20,296
ROUTES OF TRANSMISSION, AND THAT

728
00:30:20,296 --> 00:30:21,598
MEANT THAT, FOR EXAMPLE, THERE

729
00:30:21,598 --> 00:30:24,901
HAVE BEEN A NARRATIVE ABOUT TWO

730
00:30:24,901 --> 00:30:26,436
PATIENTS WHO THERE WAS THE SAME

731
00:30:26,436 --> 00:30:28,605
WOUND CART BEING USED ON THE TWO

732
00:30:28,605 --> 00:30:31,574
SUBJECTS, BUT WHEN WE SAW THEM

733
00:30:31,574 --> 00:30:33,943
IN THE GENOMIC DATA, ONE OF THEM

734
00:30:33,943 --> 00:30:35,945
WAS LIKE PATIENT 9 AND THE OTHER

735
00:30:35,945 --> 00:30:37,681
WAS PATIENT 16, YOU CAN SEE THEY

736
00:30:37,681 --> 00:30:40,383
WERE ON TWO DIFFERENT PARTS OF

737
00:30:40,383 --> 00:30:42,185
THE TRANSMISSION TREE, AND WE

738
00:30:42,185 --> 00:30:43,953
DIDN'T NEED TO CONTINUE TO LOOK

739
00:30:43,953 --> 00:30:47,057
FOR WHAT WERE THE LINKS.

740
00:30:47,057 --> 00:30:50,760
SO, WITH THIS DATA WE WERE ABLE

741
00:30:50,760 --> 00:30:51,928
TO STOP THE TRANSMISSION.

742
00:30:51,928 --> 00:30:54,497
SO NOW I WANT TO TALK ABOUT THE

743
00:30:54,497 --> 00:30:55,899
FINAL VIGNETTE WHICH IS REALLY

744
00:30:55,899 --> 00:31:00,937
ABOUT NEW WORK THAT WE'VE BEEN

745
00:31:00,937 --> 00:31:06,543
DOING ON CANDIDA AURIS COMBINING

746
00:31:06,543 --> 00:31:08,545
SKIN MICROBIOME AND RESISTANCE,

747
00:31:08,545 --> 00:31:16,786
WORKING WITH OUR COLLEAGUES AT

748
00:31:16,786 --> 00:31:18,488
THE CDC.

749
00:31:18,488 --> 00:31:23,493
SO CANDIDA AURIS IS FUNGAL

750
00:31:23,493 --> 00:31:25,261
PATHOGEN, THAT IS AGAIN CDC IS

751
00:31:25,261 --> 00:31:29,232
RATING THIS AS ONE OF ITS MOST

752
00:31:29,232 --> 00:31:31,234
URGENT THREATS BECAUSE CANDIDA

753
00:31:31,234 --> 00:31:32,635
AURIS ALSO HAS EVOLVED

754
00:31:32,635 --> 00:31:37,040
RESISTANCE TO ALL KNOWN

755
00:31:37,040 --> 00:31:37,373
ANTI-FUNGALS.

756
00:31:37,373 --> 00:31:40,110
IF YOU THINK THERE'S A PROBLEM

757
00:31:40,110 --> 00:31:45,248
WITH DEVELOPMENT OF ANTIBIOTICS,

758
00:31:45,248 --> 00:31:47,617
ANTI-FUNGALS ARE HARDER BECAUSE

759
00:31:47,617 --> 00:31:49,119
THEY ARE EUKARYOTES, HARD TO GET

760
00:31:49,119 --> 00:31:51,588
IN THE SPACE TOXIC TO FUNGI AND

761
00:31:51,588 --> 00:31:54,958
NOT TOXIC TO THE HUMAN.

762
00:31:54,958 --> 00:31:58,762
CANDIDA AURIS IS AN ENIGMA.

763
00:31:58,762 --> 00:32:01,564
IT WAS NOT KNOWN BEFORE 2009,

764
00:32:01,564 --> 00:32:08,004
AND SINCE THEN FOUR CLADES OF

765
00:32:08,004 --> 00:32:09,339
CANDIDA AURIS HAVE BEEN

766
00:32:09,339 --> 00:32:10,173
RECOGNIZED, FOUR REGIONS OF THE

767
00:32:10,173 --> 00:32:11,808
GLOBE, BUT THEY HAVE ALL NOW

768
00:32:11,808 --> 00:32:14,711
COME TO THE UNITED STATES AND

769
00:32:14,711 --> 00:32:21,117
ARE CAUSING OUTBREAKS IN NURSING

770
00:32:21,117 --> 00:32:21,584
HOMES.

771
00:32:21,584 --> 00:32:23,453
AND WHY ARE THESE CANDIDA AURIS

772
00:32:23,453 --> 00:32:27,590
OUTBREAKS  HAPPENING IN NURSING

773
00:32:27,590 --> 00:32:28,925
HOMES AND HOW DOES IT CONNECT TO

774
00:32:28,925 --> 00:32:30,193
THE SKIN MICROBIOME?

775
00:32:30,193 --> 00:32:31,661
THE SKIN IS CONSTANTLY TURNING

776
00:32:31,661 --> 00:32:31,961
OVER.

777
00:32:31,961 --> 00:32:34,230
SO WE SHED THE ENTIRE LAYER OF

778
00:32:34,230 --> 00:32:35,932
OUR SKIN EVERY TWO TO FOUR

779
00:32:35,932 --> 00:32:36,132
WEEKS.

780
00:32:36,132 --> 00:32:37,967
AND YOU'RE GOING TO SEE THAT AS

781
00:32:37,967 --> 00:32:40,170
THOSE DUST BUNNIES AROUND YOUR

782
00:32:40,170 --> 00:32:40,570
HOUSE.

783
00:32:40,570 --> 00:32:42,005
SO, WE'RE SHEDDING FIVE TIMES

784
00:32:42,005 --> 00:32:43,573
TEN TO THE EIGHTH HUMAN CELLS

785
00:32:43,573 --> 00:32:45,842
PER DAY, BUT THAT MEANS WE'RE

786
00:32:45,842 --> 00:32:48,578
SHEDDING, YOU KNOW, TEN TO THE

787
00:32:48,578 --> 00:32:51,281
SEVENTH MICROBES PER DAY.

788
00:32:51,281 --> 00:32:53,817
FOR THE MICROBES THAT CAN

789
00:32:53,817 --> 00:33:01,457
PERSIST IN THE ENVIRONMENT, LIKE

790
00:33:01,457 --> 00:33:03,493
ASENNICDOBACTER OR CANDIDA

791
00:33:03,493 --> 00:33:05,061
AURIS, IT CAN PERSIST AS VIABLE,

792
00:33:05,061 --> 00:33:11,668
THAT MAKES IT DIFFICULT TO

793
00:33:11,668 --> 00:33:13,369
CONTROL THESE OUTBREAKS IN

794
00:33:13,369 --> 00:33:14,504
CONGREGANT SETTINGS, I MEAN

795
00:33:14,504 --> 00:33:14,971
NURSING HOMES.

796
00:33:14,971 --> 00:33:17,907
WE'RE NOT SEEING THIS IN THE

797
00:33:17,907 --> 00:33:19,142
HEALTHY POPULATIONS, BUT IN

798
00:33:19,142 --> 00:33:22,946
THESE SICK OR OLDER PATIENTS

799
00:33:22,946 --> 00:33:23,913
WE'RE SEEING CANDIDA AURIS

800
00:33:23,913 --> 00:33:27,350
COLONIZE ONE RESIDENT IN THE

801
00:33:27,350 --> 00:33:29,185
NURSING HOME, THEM BEING

802
00:33:29,185 --> 00:33:31,387
BASICALLY RUNNING THROUGH THE

803
00:33:31,387 --> 00:33:32,822
ENTIRE NURSING HOME, UNLESS

804
00:33:32,822 --> 00:33:34,824
CASES ARE REALLY RECOGNIZED

805
00:33:34,824 --> 00:33:38,061
EARLY ON AND ISOLATED.

806
00:33:38,061 --> 00:33:41,497
SO, IN A STUDY THAT I'VE BEEN

807
00:33:41,497 --> 00:33:43,199
VERY FORTUNATE TO CARRY OUT WITH

808
00:33:43,199 --> 00:33:46,236
DR. MARY HAYDEN AND MY

809
00:33:46,236 --> 00:33:48,004
COLLEAGUES AT THE CDC, WE SET UP

810
00:33:48,004 --> 00:33:53,243
A STUDY TO LOOK AT RESIDENTS IN

811
00:33:53,243 --> 00:33:54,110
NURSING HOMES TO TRAP

812
00:33:54,110 --> 00:33:55,211
TRANSMISSION OF CANDIDA AURIS.

813
00:33:55,211 --> 00:34:00,250
SOME OF THAT INITIAL WORK HAS

814
00:34:00,250 --> 00:34:01,551
BEEN PUBLISHED, ABOUT HOW

815
00:34:01,551 --> 00:34:02,685
DIFFICULT IT IS TO DETECT

816
00:34:02,685 --> 00:34:04,487
CANDIDA AURIS BECAUSE IT CAN

817
00:34:04,487 --> 00:34:06,856
COLONIZE ANY PART OF THE SKIN,

818
00:34:06,856 --> 00:34:09,459
WHICH MAKES INFECTION CONTROL

819
00:34:09,459 --> 00:34:10,793
VERY HARD.

820
00:34:10,793 --> 00:34:14,831
BUT OUR LATEST WORK HAS BEEN

821
00:34:14,831 --> 00:34:16,966
LOOKING AT THE AGGREGATE OF WHAT

822
00:34:16,966 --> 00:34:18,334
IS COLONIZING THE SKIN.

823
00:34:18,334 --> 00:34:20,336
SO WE WENT IN THIS LOOKING AT

824
00:34:20,336 --> 00:34:23,072
THE SKIN AS WE WENT TO LOOK FOR

825
00:34:23,072 --> 00:34:25,875
CANDIDA AURIS BUT I WANT TO TALK

826
00:34:25,875 --> 00:34:26,409
ABOUT A SMALL TECHNOLOGY

827
00:34:26,409 --> 00:34:28,745
REVOLUTION, THIS WILL BE THE

828
00:34:28,745 --> 00:34:31,147
LAST PART, AND HOW THAT'S

829
00:34:31,147 --> 00:34:34,017
CHANGING THINGS FOR US.

830
00:34:34,017 --> 00:34:36,185
SO, I'M GOING IN AND LOOKING

831
00:34:36,185 --> 00:34:39,022
AT -- I'M LOOKING FOR CANDIDA

832
00:34:39,022 --> 00:34:39,489
AURIS.

833
00:34:39,489 --> 00:34:41,925
WHEN WE WANT TO STUDY SOMEONE'S

834
00:34:41,925 --> 00:34:45,061
SKIN MICROBIOME AND SEE, YOU

835
00:34:45,061 --> 00:34:45,962
KNOW, WHAT STRAINS OF CANDIDA

836
00:34:45,962 --> 00:34:47,997
AURIS, HOW MUCH THEY HAVE ON THE

837
00:34:47,997 --> 00:34:50,533
SKIN WE SWAB THE SKIN AND WE

838
00:34:50,533 --> 00:34:52,535
JUST PUT THAT SAMPLE -- WE CAN

839
00:34:52,535 --> 00:34:54,971
CULTURE BUT WE WANT TO LOOK AT

840
00:34:54,971 --> 00:34:57,774
THE STRAIN LEVEL VARIATION, WE

841
00:34:57,774 --> 00:34:59,442
TAKE THAT SAMPLE, CLINICAL

842
00:34:59,442 --> 00:35:02,679
SAMPLE, PUT IT STRAIGHT INTO THE

843
00:35:02,679 --> 00:35:04,180
ILLUMINA SEQUENCER.

844
00:35:04,180 --> 00:35:06,749
FROM THAT, WE CAN REASSEMBLE THE

845
00:35:06,749 --> 00:35:07,850
CANDIDA AURIS GENOME.

846
00:35:07,850 --> 00:35:10,486
THIS HAS REALLY BEEN ACTUALLY

847
00:35:10,486 --> 00:35:13,489
THE REVOLUTION OF MY LAB, WHERE

848
00:35:13,489 --> 00:35:16,159
FROM A COMPLEX SAMPLE OF THE

849
00:35:16,159 --> 00:35:20,163
SKIN, I CAN THEN REASSEMBLE THE

850
00:35:20,163 --> 00:35:23,599
ENTIRE GENOME OF CANDIDA AURIS

851
00:35:23,599 --> 00:35:25,234
AND OTHER BACTERIAL, FUNGAL, AND

852
00:35:25,234 --> 00:35:26,536
VIRAL SPECIES THAT ARE.  THERE

853
00:35:26,536 --> 00:35:29,138
HOW I THINK ABOUT THIS IS THAT I

854
00:35:29,138 --> 00:35:31,441
THINK IT'S CLEAR TO EVERYONE HOW

855
00:35:31,441 --> 00:35:33,977
YOU ALL SEQUENCE -- MANY OF YOU

856
00:35:33,977 --> 00:35:35,078
SEQUENCE MICROBIAL GENOMES AND

857
00:35:35,078 --> 00:35:37,180
KNOW YOU GET THESE READS AND

858
00:35:37,180 --> 00:35:39,449
ASSEMBLE THEM AND GET A GENOME.

859
00:35:39,449 --> 00:35:42,285
THAT'S LIKE IF I JUST GAVE

860
00:35:42,285 --> 00:35:44,253
YOU -- I HAVE PUT ALL THE PIECES

861
00:35:44,253 --> 00:35:47,090
FOR A PUZZLE INTO A BOX, AND YOU

862
00:35:47,090 --> 00:35:49,425
USE THAT TO ASSEMBLE THE GENOME.

863
00:35:49,425 --> 00:35:51,160
BUT WHAT I'M TRYING TO DESCRIBE

864
00:35:51,160 --> 00:35:55,531
TO YOU HERE WITH SHOTGUN

865
00:35:55,531 --> 00:35:58,067
METAGENOMICS TAKEN FROM THE SKIN

866
00:35:58,067 --> 00:35:59,769
SAMPLE DIRECTLY I HAVE ONE BOX

867
00:35:59,769 --> 00:36:01,571
WITH MULTIPLE PUZZLES IN IT,

868
00:36:01,571 --> 00:36:05,608
RIGHT? 

869
00:36:05,608 --> 00:36:07,510
YOU'LL HAVE THE COMMENSAL

870
00:36:07,510 --> 00:36:08,411
STAPHYLOCOCCUS AND CUDO BACTERIA

871
00:36:08,411 --> 00:36:09,545
AND CANDIDA AURIS.

872
00:36:09,545 --> 00:36:11,347
I PUT THESE TOGETHER IN ONE BOX

873
00:36:11,347 --> 00:36:16,185
AND I SAY CAN YOU ASSEMBLE THEM

874
00:36:16,185 --> 00:36:22,458
INTO GENOMES TO LOOK AT

875
00:36:22,458 --> 00:36:22,925
TRANSMISSION NETWORKS.

876
00:36:22,925 --> 00:36:23,793
THIS IS MORE EFFICIENT THAN

877
00:36:23,793 --> 00:36:32,969
PREVIOUSLY WHERE I JUST LOOKED

878
00:36:32,969 --> 00:36:34,470
AT KLEB PNEUMO, FROM ONE AT A

879
00:36:34,470 --> 00:36:34,670
TIME.

880
00:36:34,670 --> 00:36:36,806
WHEN YOU THINK IF I DID GIVE YOU

881
00:36:36,806 --> 00:36:39,742
THESE PUZZLES, YOU WOULD SORT

882
00:36:39,742 --> 00:36:41,444
THE PIECES INTO EITHER THE

883
00:36:41,444 --> 00:36:43,846
COLORS YOU SAW OR THE SIZE OF

884
00:36:43,846 --> 00:36:47,884
THE PUZZLE PIECES OR MAYBE SOME,

885
00:36:47,884 --> 00:36:49,585
YOU KNOW, DIFFERENT MARKINGS

886
00:36:49,585 --> 00:36:51,554
THAT SOME HAVE, SMALLER PIECES,

887
00:36:51,554 --> 00:36:52,321
LARGER PIECES.

888
00:36:52,321 --> 00:36:56,526
THAT'S EXACTLY WHAT WE'RE DOING

889
00:36:56,526 --> 00:36:58,795
ALSO, LOOKING AT TETRA

890
00:36:58,795 --> 00:37:00,463
NUCLEOSIDE, GC CONTENT, SORTING

891
00:37:00,463 --> 00:37:00,797
INTO PUZZLES.

892
00:37:00,797 --> 00:37:04,167
I WAS EXPECTING TO SORT OF LOOK

893
00:37:04,167 --> 00:37:06,469
AT FRIENDS AND FOES AND CANDIDA

894
00:37:06,469 --> 00:37:09,272
AURIS, AND INSTEAD WHEN WE

895
00:37:09,272 --> 00:37:11,974
ASSEMBLED ALL OF THE GENOMES

896
00:37:11,974 --> 00:37:14,243
WHAT WE FOUND WAS CANDIDA AURIS,

897
00:37:14,243 --> 00:37:16,412
AND FROM THE SKIN OF THESE

898
00:37:16,412 --> 00:37:19,248
RESIDENTS OF THE NURSING HOME

899
00:37:19,248 --> 00:37:20,616
FOUND A CLONAL SPREAD OF CANDIDA

900
00:37:20,616 --> 00:37:22,652
AURIS, IN ADDITION THE OTHER

901
00:37:22,652 --> 00:37:24,120
GENOMES THAT CAME OUT OF THE

902
00:37:24,120 --> 00:37:27,290
SKIN OF THESE PATIENTS IN THE

903
00:37:27,290 --> 00:37:36,566
NURSING HOMES WAS E. COLI, STAPH

904
00:37:36,566 --> 00:37:37,300
AUREUS, PSEUDOMONAS AERUGINOSA,

905
00:37:37,300 --> 00:37:40,436
THE FULL ESCAPE OF ALL OF THE

906
00:37:40,436 --> 00:37:43,172
BACTERIA WE CONSIDER AND ARE

907
00:37:43,172 --> 00:37:49,812
MOST CONCERNED ABOUT WITH

908
00:37:49,812 --> 00:37:51,214
MULTI-DRUG RESISTANCE.

909
00:37:51,214 --> 00:37:52,181
WE ASSEMBLED FULL GENESOME FROM

910
00:37:52,181 --> 00:37:55,017
THE SKIN OF THESE PATIENTS AND

911
00:37:55,017 --> 00:37:59,155
ARE TRACKING, WE WERE TRYING TO 

912
00:37:59,155 --> 00:38:02,091
TRACK A CANDIDA AURIS OUTBREAK

913
00:38:02,091 --> 00:38:02,658
BUT INSTEAD TRACKING

914
00:38:02,658 --> 00:38:06,329
SIMULTANEOUSY SPREAD OF CANDIDA

915
00:38:06,329 --> 00:38:11,868
AURIS, KLEBSIELLA PNEUMO, E.

916
00:38:11,868 --> 00:38:15,805
COLI, AND STAPH AND PSEUDOMONAS,

917
00:38:15,805 --> 00:38:23,446
HAVING THE SPREAD OF A LOT MORE

918
00:38:23,446 --> 00:38:24,280
MULTIDRUG-RESISTANT ORGANISMS

919
00:38:24,280 --> 00:38:25,348
THAN HAD BEEN PREVIOUSLY

920
00:38:25,348 --> 00:38:25,648
APPRECIATED.

921
00:38:25,648 --> 00:38:27,817
AND THIS BRINGS ME TO MY FINAL

922
00:38:27,817 --> 00:38:31,420
POINT, WHICH IS, YOU KNOW, AS WE

923
00:38:31,420 --> 00:38:32,655
THINK ABOUT ANTIBIOTIC

924
00:38:32,655 --> 00:38:33,656
STEWARDSHIP IN HOSPITALS, AND SO

925
00:38:33,656 --> 00:38:36,192
ON, ONE OF THE THINGS THAT COVID

926
00:38:36,192 --> 00:38:39,028
HAS BROUGHT TO BEAR IS THAT

927
00:38:39,028 --> 00:38:41,831
NURSING HOMES ARE PART OF OUR

928
00:38:41,831 --> 00:38:43,733
HEALTH CARE ECOSYSTEMS, AND THEY

929
00:38:43,733 --> 00:38:45,501
DON'T HAVE THE SAME LEVELS OF

930
00:38:45,501 --> 00:38:48,104
INFECTION CONTROL BUT WE NEED TO

931
00:38:48,104 --> 00:38:49,939
START THINKING ABOUT THEM AS

932
00:38:49,939 --> 00:38:52,475
ALSO PLACES WHERE WE NEED

933
00:38:52,475 --> 00:38:55,711
ENHANCED INFECTION CONTROL AND 

934
00:38:55,711 --> 00:38:56,946
CARE FOR THESE PATIENTS.

935
00:38:56,946 --> 00:38:59,949
SO WITH THAT, I AM HAVING

936
00:38:59,949 --> 00:39:03,052
TROUBLE NOW SOMEHOW -- OKAY.

937
00:39:03,052 --> 00:39:04,554
MY OVERALL CONCLUSIONS, GENOMIC

938
00:39:04,554 --> 00:39:06,088
SEQUENCING MAKES THE INVISIBLE

939
00:39:06,088 --> 00:39:10,927
WORLD OF MICROBES VISIBLE AND 

940
00:39:10,927 --> 00:39:12,962
IDENTIFIABLE AND I HOPE WE'LL

941
00:39:12,962 --> 00:39:16,933
REVOLUTIONIZE WHAT IT MEANS TO

942
00:39:16,933 --> 00:39:19,202
BE HUMAN, HEALTHY, PROVIDE

943
00:39:19,202 --> 00:39:23,239
INSIGHTS TO FIGHT MICROBIAL FOES

944
00:39:23,239 --> 00:39:24,574
AND CULTIVATE MICROBIAL FRIENDS.

945
00:39:24,574 --> 00:39:26,075
I'D LIKE TO ACKNOWLEDGE MY

946
00:39:26,075 --> 00:39:26,642
COLLABORATORS.

947
00:39:26,642 --> 00:39:27,743
IT'S BEEN THE PLEASURE AND HONOR

948
00:39:27,743 --> 00:39:31,547
OF MY LIFE TO WORK AT NIH, AND

949
00:39:31,547 --> 00:39:33,316
ALL OF THESE STUDIES WERE MADE

950
00:39:33,316 --> 00:39:35,718
POSSIBLE BECAUSE OF THESE

951
00:39:35,718 --> 00:39:37,553
COLLABORATIONS, IN PARTICULAR

952
00:39:37,553 --> 00:39:38,654
THE LONG-TERM COLLABORATIONS

953
00:39:38,654 --> 00:39:45,761
WITH DR. HEIDI KONG, TAR A

954
00:39:45,761 --> 00:39:48,864
PALMORE, YASMINE BELKAID, NIH 

955
00:39:48,864 --> 00:39:51,334
AND CDC AND LARGER COMMUNITY,

956
00:39:51,334 --> 00:39:58,040
ESPECIALLY SHOUT OUT TO MY LAB,

957
00:39:58,040 --> 00:40:01,110
FROM 2011 AND 2013, AND REALLY

958
00:40:01,110 --> 00:40:02,612
THE PERMANENT MEMBERS OF THE LAB

959
00:40:02,612 --> 00:40:05,615
WHO ARE SHOWN IN BOTH OF THESE

960
00:40:05,615 --> 00:40:09,418
PICTURES, GREAT MENTORS I'VE HAD

961
00:40:09,418 --> 00:40:11,854
AT NHGRI AND NIH, AND MY FAMILY.

962
00:40:11,854 --> 00:40:14,523
SO THANK YOU ALL.

963
00:40:14,523 --> 00:40:17,159
AND I'M HAPPY TO TAKE QUESTIONS.

964
00:40:17,159 --> 00:40:27,637
SORRY I COULDN'T JOIN YOU IN

965
00:40:31,841 --> 00:40:32,008
PERSON.

966
00:40:32,008 --> 00:40:32,275
[APPLAUSE]

967
00:40:32,275 --> 00:40:35,778
>> WE'RE GOING TO MOVE ON,
I

968
00:40:35,778 --> 00:40:36,746
CONVEY GET WELL WISHES AND THANK

969
00:40:36,746 --> 00:40:40,016
YOU FOR BEING PART OF THE

970
00:40:40,016 --> 00:40:40,283
SYMPOSIUM.

971
00:40:40,283 --> 00:40:45,554
NEXT SPEAKER IS DR. MICHAEL

972
00:40:45,554 --> 00:40:47,923
LICHTEN, SCIENTIST EMERITUS AT

973
00:40:47,923 --> 00:40:48,391
NCI.

974
00:40:48,391 --> 00:40:52,328
HE'S AN EXPERT ON MEIOSIS, THE

975
00:40:52,328 --> 00:40:54,797
PROCESS BY WHICH GAMETES ARE

976
00:40:54,797 --> 00:40:58,034
FORMED, AND PARTICULARLY ON THE

977
00:40:58,034 --> 00:41:01,304
PROCESS OF EXCHANGE OF CHROMATIN

978
00:41:01,304 --> 00:41:03,272
DURING MEIOSIS, AND WHAT HAP%ES

979
00:41:03,272 --> 00:41:07,343
WHEN THAT PROCESS GOES AWRY.

980
00:41:07,343 --> 00:41:11,781
HE USES A YEAST MODEL TO LOOK AT

981
00:41:11,781 --> 00:41:12,415
THESE RECOMBINATION EVENTS, AND

982
00:41:12,415 --> 00:41:14,884
I'M GOING TO TURN THE PODIUM 

983
00:41:14,884 --> 00:41:21,724
OVER TO DR. LICHTEN. 

984
00:41:21,724 --> 00:41:24,026
>> THANK YOU, NINA.

985
00:41:24,026 --> 00:41:29,865
YOU JUST BLEW 90% OF MY 

986
00:41:29,865 --> 00:41:30,299
INTRODUCTORY SLIDES.

987
00:41:30,299 --> 00:41:33,636
[LAUGHTER] 

988
00:41:33,636 --> 00:41:33,836
OKAY.

989
00:41:33,836 --> 00:41:44,213
OOPS, THE OTHER WAY. 

990
00:41:46,449 --> 00:41:50,019
HERE WE GO.

991
00:41:50,019 --> 00:41:51,454
YOU WILL NOTICE A CERTAIN

992
00:41:51,454 --> 00:41:53,489
REPETITIVE NATURE TO THE TITLES

993
00:41:53,489 --> 00:41:53,956
OF THESE TALKS.

994
00:41:53,956 --> 00:42:00,730
I DON'T KNOW HOW LONG IT'S GOING

995
00:42:00,730 --> 00:42:03,933
TO EXTEND THROUGH THE SYMPOSIUM.

996
00:42:03,933 --> 00:42:07,002
I'M INTERESTED IN MEIOSIS AND

997
00:42:07,002 --> 00:42:07,737
RECOMBINATION, BUT IN PARTICULAR

998
00:42:07,737 --> 00:42:13,109
IN THE REPAIR OF DNA BREAKS, AND

999
00:42:13,109 --> 00:42:17,046
THE MECHANISTIC INSIGHTS WE CAN

1000
00:42:17,046 --> 00:42:17,613
GAIN.

1001
00:42:17,613 --> 00:42:19,882
SO, YOU KNOW EACH ONE OF US IN

1002
00:42:19,882 --> 00:42:21,650
EVERY ONE OF OUR CELLS, EVERY

1003
00:42:21,650 --> 00:42:26,188
ONE OF OUR CELLS THAT DIVIDES,

1004
00:42:26,188 --> 00:42:27,656
UNDERGOES DOZENS OF

1005
00:42:27,656 --> 00:42:32,628
DOUBLE-STRAND BREAKS, EVERY CELL

1006
00:42:32,628 --> 00:42:32,895
CYCLE.

1007
00:42:32,895 --> 00:42:34,697
AND THIS IS -- IN FACT, IF YOU

1008
00:42:34,697 --> 00:42:37,299
LOOK AT THE NUMBER THROUGHOUT

1009
00:42:37,299 --> 00:42:40,302
THE ANIMAL KINGDOM, IT'S WITHIN

1010
00:42:40,302 --> 00:42:41,904
A COUPLE OF FACTORS OF TWO OF

1011
00:42:41,904 --> 00:42:42,638
EACH OTHER.

1012
00:42:42,638 --> 00:42:49,345
SO THIS SEEMS TO BE PRETTY MUCH

1013
00:42:49,345 --> 00:42:52,114
UNIVERSAL AMONG EUKARYOTES.

1014
00:42:52,114 --> 00:42:55,484
INACCURATE REPAIR, CELLS ARE

1015
00:42:55,484 --> 00:42:56,819
DEPENDENT ON ACCURATE REPAIR,

1016
00:42:56,819 --> 00:42:59,622
INACCURATE REPAIR CAN LEAD TO

1017
00:42:59,622 --> 00:43:01,791
MUTATIONS AND CHROMOSOME

1018
00:43:01,791 --> 00:43:02,458
REARRANGEMENT, AND IN PARTICULAR

1019
00:43:02,458 --> 00:43:03,926
I LIKE TO SHOW THIS PICTURE

1020
00:43:03,926 --> 00:43:09,398
WHICH IS THE FACE OF THE ENEMY. 

1021
00:43:09,398 --> 00:43:16,138
THIS IS A SPECTRAL YOUR CARE  --

1022
00:43:16,138 --> 00:43:17,640
KARYOTYPE OF A BLADDER CELL BY

1023
00:43:17,640 --> 00:43:18,340
THOMAS REIDED A NCI.

1024
00:43:18,340 --> 00:43:23,145
I THINK THERE ARE OVER A DOZEN

1025
00:43:23,145 --> 00:43:24,480
CHROMOSOME REARRANGEMENTS IN

1026
00:43:24,480 --> 00:43:26,449
THIS SOLID TUMOR, AND EACH ONE

1027
00:43:26,449 --> 00:43:27,883
OF THOSE WAS A DOUBLE STRAND

1028
00:43:27,883 --> 00:43:30,820
BREAK THAT AT ONE TIME WAS

1029
00:43:30,820 --> 00:43:32,922
REPAIRED INCORRECTLY.

1030
00:43:32,922 --> 00:43:35,491
SO, THIS IS OF RELEVANCE FOR A

1031
00:43:35,491 --> 00:43:40,162
NUMBER OF THINGS BUT CERTAINLY 

1032
00:43:40,162 --> 00:43:41,430
IN ETIOLOGY OF CANCER.

1033
00:43:41,430 --> 00:43:47,203
SO BREAKS CAN BE REPAIRED BY TWO

1034
00:43:47,203 --> 00:43:48,437
MECHANISMS, NON-HOMOLOGOUS

1035
00:43:48,437 --> 00:43:50,072
JOINING, BASICALLY JUST BASHING

1036
00:43:50,072 --> 00:43:51,307
AND LIGATION, AND THAT'S WHAT

1037
00:43:51,307 --> 00:43:55,845
GIVES YOU PROBLEMS LIKE THIS.

1038
00:43:55,845 --> 00:43:59,715
OR BY HOMOLOGOUS RECOMBINATION, 

1039
00:43:59,715 --> 00:44:04,453
WHICH PRESERVES GENOME CONTENT.

1040
00:44:04,453 --> 00:44:06,388
BUT THIS IS -- DOUBLE-STRAND

1041
00:44:06,388 --> 00:44:08,457
BREAKS DON'T GO AWAY WITH THE

1042
00:44:08,457 --> 00:44:08,991
THOUGHT THAT DOUBLE-STRAND

1043
00:44:08,991 --> 00:44:10,226
BREAKS WITH JUST A CHALLENGE TO

1044
00:44:10,226 --> 00:44:13,062
THE CELL.

1045
00:44:13,062 --> 00:44:15,364
IN FACT, OUR CELLS AND CELLS OF

1046
00:44:15,364 --> 00:44:20,369
OTHER ORGANISMS MAKE WILLFUL USE

1047
00:44:20,369 --> 00:44:21,303
OF DOUBLE-STRAND BREAKS, WILLFUL

1048
00:44:21,303 --> 00:44:23,839
DAMAGING OF GENOMES TO CARRY OUT

1049
00:44:23,839 --> 00:44:25,441
VARIOUS FUNCTIONS.

1050
00:44:25,441 --> 00:44:27,877
ONE MIGHT WONDER WHY, BUT THE

1051
00:44:27,877 --> 00:44:30,513
FACT IS EVERY CELL THAT DIVIDES

1052
00:44:30,513 --> 00:44:35,918
IN OUR BODY IS REPAIRING 50

1053
00:44:35,918 --> 00:44:36,352
DOUBLE-STRAND BREAKS

1054
00:44:36,352 --> 00:44:42,658
EFFICIENTLY, SO WHAT'S A FEW

1055
00:44:42,658 --> 00:44:43,959
MORE?

1056
00:44:43,959 --> 00:44:45,661
SO, AMONG THOSE IS SWITCHING IN

1057
00:44:45,661 --> 00:44:49,565
FUNGI WITH ONE BREAK OR SO PER

1058
00:44:49,565 --> 00:44:53,035
CELL, EVERY MATING TYPE SWITCH,

1059
00:44:53,035 --> 00:44:56,972
GENERATION OF IMMUNOGLOBULIN

1060
00:44:56,972 --> 00:45:00,609
DEFICIENCY REPAIRED BY

1061
00:45:00,609 --> 00:45:02,144
HOMOLOGOUS RECOMBINATION IN VDJ

1062
00:45:02,144 --> 00:45:04,179
RECOMBINATION, CLASS SWITCH

1063
00:45:04,179 --> 00:45:08,450
RECOMBINATION, ET CETERA,

1064
00:45:08,450 --> 00:45:09,251
REPAIRING BY NON-HOMOLOGOUS END

1065
00:45:09,251 --> 00:45:10,519
JOINING BY A COUPLE BREAKS PER

1066
00:45:10,519 --> 00:45:10,920
CELL.

1067
00:45:10,920 --> 00:45:14,290
THEN YOU GET TO MEIOSIS, WHERE

1068
00:45:14,290 --> 00:45:17,660
THERE ARE ON THE ORDER -- THERE

1069
00:45:17,660 --> 00:45:19,295
ARE HUNDREDS OF HUNDREDS OF

1070
00:45:19,295 --> 00:45:20,729
BREAKS FORMED DURING THE PROCESS

1071
00:45:20,729 --> 00:45:21,830
OF MEIOSIS.

1072
00:45:21,830 --> 00:45:24,233
WHEN I WAS LOOKING FOR A MODEL

1073
00:45:24,233 --> 00:45:28,737
SYSTEM TO STUDY DNA DAMAGE

1074
00:45:28,737 --> 00:45:35,210
REPAIR, AND HOMOLOGOUS

1075
00:45:35,210 --> 00:45:36,078
RECOMBINATION, I FOCUSED ON

1076
00:45:36,078 --> 00:45:36,312
MEIOSIS

1077
00:45:36,312 --> 00:45:38,147
WHAT IS MEIOSIS?

1078
00:45:38,147 --> 00:45:40,282
MEIOSIS IS THE PROCESS, EXCUSE

1079
00:45:40,282 --> 00:45:44,887
ME.

1080
00:45:44,887 --> 00:45:51,827
MEIOSIS IS THE PROCESS BY WHICH

1081
00:45:51,827 --> 00:45:55,965
A DIPLOID GENOME IS DIVIDED INTO

1082
00:45:55,965 --> 00:45:57,933
FOUR HAPLOID GAMETES, SPERM --

1083
00:45:57,933 --> 00:46:00,035
EITHER SPERM OR THE EGG, AND IN

1084
00:46:00,035 --> 00:46:04,773
DOING SO IT GOES FIRST THROUGH A

1085
00:46:04,773 --> 00:46:07,176
ROUND OF REPLICATION, AND THEN

1086
00:46:07,176 --> 00:46:09,445
THE CELL DOES SOMETHING THAT --

1087
00:46:09,445 --> 00:46:11,647
EXCUSE ME, BEAR WITH ME FOR A

1088
00:46:11,647 --> 00:46:15,117
MINUTE, I'M GOING TO TRY AND

1089
00:46:15,117 --> 00:46:17,219
TURN ON THIS LASER POINTER.

1090
00:46:17,219 --> 00:46:20,189
THERE WE GO.

1091
00:46:20,189 --> 00:46:20,456
OKAY.

1092
00:46:20,456 --> 00:46:22,891
IT DOES SOMETHING THAT IT DOES

1093
00:46:22,891 --> 00:46:26,495
NO OTHER TIME IN THE LIFE CYCLE

1094
00:46:26,495 --> 00:46:27,429
OF THE ORGANISM.

1095
00:46:27,429 --> 00:46:29,999
RATHER THAN SEPARATE THE TWO

1096
00:46:29,999 --> 00:46:32,635
SISTER COPIES OF EACH

1097
00:46:32,635 --> 00:46:35,070
CHROMOSOME, IT GOES THROUGH AN

1098
00:46:35,070 --> 00:46:37,039
ELABORATE PROCESS OF TWO NUCLEAR

1099
00:46:37,039 --> 00:46:40,409
DIVISIONS, ENDING UP WITH FOUR

1100
00:46:40,409 --> 00:46:42,344
HAPLOID GAMETES WHICH INVOLVES

1101
00:46:42,344 --> 00:46:45,447
THE ASSOCIATION AND PAIRING OF

1102
00:46:45,447 --> 00:46:49,151
THE TWO COPIES OF THE SAME

1103
00:46:49,151 --> 00:46:49,718
CHROMOSOME, WHICH PREVIOUSLY

1104
00:46:49,718 --> 00:46:51,120
HAVE NO RELATIONSHIP TO EACH

1105
00:46:51,120 --> 00:46:53,088
OTHER, SO THEY HAVE TO BE

1106
00:46:53,088 --> 00:46:56,592
BROUGHT TOGETHER AND STABLY

1107
00:46:56,592 --> 00:46:58,360
PAIRED.

1108
00:46:58,360 --> 00:47:00,663
AND THEN CONNECTED BY

1109
00:47:00,663 --> 00:47:02,498
CHROMOSOME, BY CROSSOVERS THAT

1110
00:47:02,498 --> 00:47:04,233
LINK THE TWO HOMOLOGOUS

1111
00:47:04,233 --> 00:47:08,637
CHROMOSOMES AND PROVIDE THE 

1112
00:47:08,637 --> 00:47:10,506
CONNECTION THAT ALLOWS THE

1113
00:47:10,506 --> 00:47:12,975
TENSION TO BE SENSED AT THE

1114
00:47:12,975 --> 00:47:15,310
FIRST DIVISION OF THE SPINDLE OF

1115
00:47:15,310 --> 00:47:16,945
THE FIRST DIVISION.

1116
00:47:16,945 --> 00:47:21,884
AND THEN THE SECOND DIVISION IS

1117
00:47:21,884 --> 00:47:24,720
A MORE MITOSIS-LIKE DIVISION.

1118
00:47:24,720 --> 00:47:26,722
SO, RECOMBINATION PLAYS A

1119
00:47:26,722 --> 00:47:30,759
CRITICAL ROLE IN BOTH THE EARLY

1120
00:47:30,759 --> 00:47:32,761
STEPS OF MEIOSIS, HOMOLOGS COME

1121
00:47:32,761 --> 00:47:35,764
TOGETHER AND ARE STABLY PAIRED,

1122
00:47:35,764 --> 00:47:37,900
AND IN COMING CROSSOVERS THAT AS

1123
00:47:37,900 --> 00:47:43,772
I SAID PROVIDE STABLE LICKS

1124
00:47:43,772 --> 00:47:45,340
BETWEEN HOMOLOGOUS CHROMOSOMES.

1125
00:47:45,340 --> 00:47:47,443
WHEN THIS GOES WRONG

1126
00:47:47,443 --> 00:47:48,510
CONSEQUENCES ARE DIRE.

1127
00:47:48,510 --> 00:47:59,054
MANY OF YOU HAVE RECOGNIZED THIS

1128
00:48:04,093 --> 00:48:05,494
CURVE, AMYPLOIDY WITH A FUNCTION

1129
00:48:05,494 --> 00:48:06,261
OF MATERNAL AGE.

1130
00:48:06,261 --> 00:48:08,764
I WANT TO POINT OUT ALL ACROSS

1131
00:48:08,764 --> 00:48:14,002
THIS CURVE, BUT IN PARTICULAR IN

1132
00:48:14,002 --> 00:48:15,771
THESE LATER STAGES WHERE

1133
00:48:15,771 --> 00:48:20,142
ANEUPLOIDY CAN BE HIGH THERE'S

1134
00:48:20,142 --> 00:48:20,776
CLOSE ASSOCIATION WITH

1135
00:48:20,776 --> 00:48:24,813
RECOMBINATION HAPPENING IN THE

1136
00:48:24,813 --> 00:48:26,648
WRONG PLACE.

1137
00:48:26,648 --> 00:48:28,684
IT'S IMPORTANT TO MAINTAIN -- TO

1138
00:48:28,684 --> 00:48:32,187
DO THIS PROBABLY IN ORDER TO

1139
00:48:32,187 --> 00:48:34,857
SEGREGATE CHROMOSOMES PROPERLY.

1140
00:48:34,857 --> 00:48:35,657
OKAY.

1141
00:48:35,657 --> 00:48:36,792
SO HOW DOES RECOMBINATION PLAY A

1142
00:48:36,792 --> 00:48:39,294
ROLE IN THIS?

1143
00:48:39,294 --> 00:48:42,598
AS I SAID, EARLY STEPS INVOLVE

1144
00:48:42,598 --> 00:48:45,868
MULTIPLE INTERACTION, MULTIPLE

1145
00:48:45,868 --> 00:48:48,237
TRANSIENT INTERACTIONS BETWEEN

1146
00:48:48,237 --> 00:48:51,740
HOMOLOGOUS CHROMOSOMES, 

1147
00:48:51,740 --> 00:48:54,376
INITIATED BY PROGRAM 

1148
00:48:54,376 --> 00:48:56,945
DOUBLE-STRAND STRAND BREAKS BY

1149
00:48:56,945 --> 00:48:58,280
SPO11 PROTEIN, HUNDREDS FORMED

1150
00:48:58,280 --> 00:49:00,082
PER GENOME.

1151
00:49:00,082 --> 00:49:01,083
NORMALLY MITOTIC CELLS SUCH

1152
00:49:01,083 --> 00:49:05,087
BREAKS WOULD BE REPAIRED BY

1153
00:49:05,087 --> 00:49:06,021
RECOMBINATION WITH SISTER

1154
00:49:06,021 --> 00:49:08,757
CHROMATID BUT IN THIS CASE USED

1155
00:49:08,757 --> 00:49:13,128
TO PAIR WITH HOMOLOGOUS, SO

1156
00:49:13,128 --> 00:49:15,297
SISTER RECOMBINATION IS

1157
00:49:15,297 --> 00:49:20,469
DISCOURAGED, RECOMBINATION WITH

1158
00:49:20,469 --> 00:49:21,570
HOMOLOGOUS CHROMOSOME IS

1159
00:49:21,570 --> 00:49:23,739
ENCOURAGED, SORTED INTO TWO

1160
00:49:23,739 --> 00:49:26,275
TYPES, MOST ARE SORTED OUT INTO

1161
00:49:26,275 --> 00:49:28,410
NON-CROSSOVERS, AND THEN A FEW

1162
00:49:28,410 --> 00:49:30,412
OF THEM ARE MATURED INTO

1163
00:49:30,412 --> 00:49:31,713
CROSSOVERS THAT, AGAIN, ARE

1164
00:49:31,713 --> 00:49:33,515
PROVIDING THE CONNECTIONS WITH

1165
00:49:33,515 --> 00:49:35,951
HOMOLOGS THAT ALLOW PROPER

1166
00:49:35,951 --> 00:49:36,885
SEGREGATION.

1167
00:49:36,885 --> 00:49:39,788
THAT ALLOW PROPER HOMOLOG

1168
00:49:39,788 --> 00:49:40,088
SEGREGATION.

1169
00:49:40,088 --> 00:49:44,126
SO, MY LAB'S BEEN INTERESTED IN

1170
00:49:44,126 --> 00:49:44,893
ALL OF THESE, MOLECULAR

1171
00:49:44,893 --> 00:49:47,896
MECHANISM OF ALL THESE STEPS

1172
00:49:47,896 --> 00:49:52,701
ALONG THIS PROCESS.

1173
00:49:52,701 --> 00:49:57,306
WE'VE STUDIED THE RULES THAT

1174
00:49:57,306 --> 00:50:01,376
DETERMINE WHERE SPO11 MAKES 

1175
00:50:01,376 --> 00:50:02,911
DOUBLE-STRAND BREAKS, STUDY

1176
00:50:02,911 --> 00:50:05,981
MECHANISMS THAT DISCOURAGE

1177
00:50:05,981 --> 00:50:07,950
SISTER CHROMATID RECOMBINATION,

1178
00:50:07,950 --> 00:50:08,784
ENCOURAGE INTERHOMOLOG

1179
00:50:08,784 --> 00:50:10,118
RECOMBINATION BUT MOST OF THE

1180
00:50:10,118 --> 00:50:12,020
FOCUS HAS BEEN ON MECHANISMS

1181
00:50:12,020 --> 00:50:12,688
THAT DETERMINE WHETHER

1182
00:50:12,688 --> 00:50:14,089
RECOMBINATION EVENT IS GOING TO

1183
00:50:14,089 --> 00:50:17,893
FORM A CROSSOVER OR

1184
00:50:17,893 --> 00:50:19,795
NON-CROSSOVER, AND THE

1185
00:50:19,795 --> 00:50:21,830
REGULATORY MECHANISMS, THE

1186
00:50:21,830 --> 00:50:23,398
ACTIVITIES THAT REGULATE WHETHER

1187
00:50:23,398 --> 00:50:25,133
ONE STEP IS -- WHETHER ONE FATE

1188
00:50:25,133 --> 00:50:28,670
IS FOLLOWED OR THE OTHER.

1189
00:50:28,670 --> 00:50:34,977
SO, AND OUR WORK HAS BEEN DONE

1190
00:50:34,977 --> 00:50:37,212
EXCLUSIVELY IN BUDDING YEAST,

1191
00:50:37,212 --> 00:50:38,814
I'LL DISPENSE WITH THIS BRAG

1192
00:50:38,814 --> 00:50:42,417
SLIDE BUT JUST TO POINT OUT THAT

1193
00:50:42,417 --> 00:50:45,187
BECAUSE YEAST IS A SINGLE-CELLED

1194
00:50:45,187 --> 00:50:47,823
ORGANISM WE CAN GROW IT IN

1195
00:50:47,823 --> 00:50:50,926
LIQUID CULTURE, WE CAN INDUCE

1196
00:50:50,926 --> 00:50:52,527
MEIOSIS SYNCHRONOUSLY AND

1197
00:50:52,527 --> 00:50:53,595
EXTRACT DNA FROM CELLS AS THEY

1198
00:50:53,595 --> 00:50:56,665
ARE GOING THROUGH THE PROCESS OF

1199
00:50:56,665 --> 00:50:58,967
RECOMBINATION, AND THEREFORE BY

1200
00:50:58,967 --> 00:50:59,968
ANALYZING THE INTERMEDIATES AND

1201
00:50:59,968 --> 00:51:03,105
PRODUCTS AMONG THE DNA THAT WE

1202
00:51:03,105 --> 00:51:04,873
ISOLATE, WE CAN MONITOR

1203
00:51:04,873 --> 00:51:06,074
RECOMBINATION BASICALLY IN REAL

1204
00:51:06,074 --> 00:51:06,275
TIME.

1205
00:51:06,275 --> 00:51:09,912
AND SO WE CAN SEE THE

1206
00:51:09,912 --> 00:51:11,680
INTERMEDIATE STEPS, WHEN THE

1207
00:51:11,680 --> 00:51:19,321
INTERMEDIATE STEPS GO TO

1208
00:51:19,321 --> 00:51:22,557
PRODUCTS, ET CETERA. 

1209
00:51:22,557 --> 00:51:26,995
UNLIKE MANY ORGANISMS

1210
00:51:26,995 --> 00:51:27,763
PARTICULARLY DURING OOGENESIS

1211
00:51:27,763 --> 00:51:28,964
RECOVER ALL FOUR HAPLOID

1212
00:51:28,964 --> 00:51:37,606
PRODUCTS AND  AND TETRADS,

1213
00:51:37,606 --> 00:51:39,107
ANALYZE PARENTAL CONTRIBUTION IN

1214
00:51:39,107 --> 00:51:44,379
EVERY RECOMBINATION EVENT.

1215
00:51:44,379 --> 00:51:46,048
FINALLY, I WOULDN'T BE UP HERE

1216
00:51:46,048 --> 00:51:51,920
IF IT WASN'T TRUE MY  MEIOSIS IS

1217
00:51:51,920 --> 00:51:59,261
SIMILAR TO MEIOSIS IN U.S. US,

1218
00:51:59,261 --> 00:52:01,530
DIRECTLY APPLICABLE TO OTHER

1219
00:52:01,530 --> 00:52:01,797
ORGANISMS.

1220
00:52:01,797 --> 00:52:02,631
THE PRINCIPLES WE'VE DISCOVERED

1221
00:52:02,631 --> 00:52:07,269
IN OUR STUDY OF THE MECHANISMS

1222
00:52:07,269 --> 00:52:09,004
OF MEIOTIC RECOMBINATION HAVE

1223
00:52:09,004 --> 00:52:11,707
OVER THE YEARS INFORMED

1224
00:52:11,707 --> 00:52:15,310
MECHANISMS OF REPAIR OF

1225
00:52:15,310 --> 00:52:16,712
DOUBLE-STRAND BREAKS AND

1226
00:52:16,712 --> 00:52:18,313
HOMOLOGOUS RECOMBINATION IN THE

1227
00:52:18,313 --> 00:52:19,381
MITOTIC CYCLE AS WELL.

1228
00:52:19,381 --> 00:52:19,715
OKAY.

1229
00:52:19,715 --> 00:52:25,320
I'M GOING TO TELL YOU ONE STORY

1230
00:52:25,320 --> 00:52:26,688
ABOUT MEIOTIC RECOMBINATION.

1231
00:52:26,688 --> 00:52:32,728
IT STARTS AROUND THE TURN OF THE

1232
00:52:32,728 --> 00:52:34,029
CENTURY WITH A POSTDOC WHO WAS

1233
00:52:34,029 --> 00:52:37,099
INTERESTED IN TRYING TO FIGURE

1234
00:52:37,099 --> 00:52:42,704
OUT THE RULES FOR NON-CROSSOVER

1235
00:52:42,704 --> 00:52:44,139
VERSUS CROSSOVER RECOMBINATION.

1236
00:52:44,139 --> 00:52:47,109
I WON'T SHOW -- I'LL SHOW A

1237
00:52:47,109 --> 00:52:51,580
LITTLE BIT OF DATA ENFORCING 

1238
00:52:51,580 --> 00:52:54,016
THIS LATER ON, BUT WHAT HE FOUND

1239
00:52:54,016 --> 00:52:57,019
OUT IN FACT AFTER ONE HAD MADE

1240
00:52:57,019 --> 00:52:59,321
AN INITIAL STRAND INVASION

1241
00:52:59,321 --> 00:53:02,491
INTERMEDIA, NOT CROSSOVERS AND

1242
00:53:02,491 --> 00:53:03,892
CROSSOVERS WERE FORMED BY

1243
00:53:03,892 --> 00:53:06,228
DISTINCT, VERY DISTINCT AND

1244
00:53:06,228 --> 00:53:07,095
DIFFERENT BIOCHEMICAL

1245
00:53:07,095 --> 00:53:07,396
MECHANISMS.

1246
00:53:07,396 --> 00:53:11,867
AND HE GOT A HINT THAT THAT WAS

1247
00:53:11,867 --> 00:53:15,871
TRUE JUST  BY LOOKING AT TIMING.

1248
00:53:15,871 --> 00:53:19,007
I'VE DRAWN A TIME LINE.

1249
00:53:19,007 --> 00:53:24,446
SO NON-CROSSOVERS APPEAR EARLY,

1250
00:53:24,446 --> 00:53:28,550
WELL BEFORE CROSSOVERS APPEAR,

1251
00:53:28,550 --> 00:53:31,753
AND WHEREAS CROSSOVERS DERIVE

1252
00:53:31,753 --> 00:53:33,121
FROM THIS INTERMEDIATE, DOUBLE

1253
00:53:33,121 --> 00:53:36,525
HOLLIDAY JUNCTION, AND THE 

1254
00:53:36,525 --> 00:53:40,162
RESOLUTION OF THIS INTERMEDIATE,

1255
00:53:40,162 --> 00:53:42,998
AND ITS PRECURSOR CALLED SINGLE

1256
00:53:42,998 --> 00:53:46,168
END INVASION INTERMEDIATE, AND

1257
00:53:46,168 --> 00:53:47,969
THIS RESOLUTION IS DEPENDENT

1258
00:53:47,969 --> 00:53:53,408
UPON -- ACTIVATED BY IN YEAST

1259
00:53:53,408 --> 00:53:59,881
THE CDC5 POLO KINASE UNDER

1260
00:53:59,881 --> 00:54:06,288
CONTROL OF THE NDT80  TRANSCRIPT

1261
00:54:06,288 --> 00:54:06,521
FACTOR.

1262
00:54:06,521 --> 00:54:10,926
IF ONE MUTATED EITHER OF THESE

1263
00:54:10,926 --> 00:54:14,296
PROTEINS, THESE GENES, ONE

1264
00:54:14,296 --> 00:54:18,100
TERMINATED MEIOSIS WITH MATURE

1265
00:54:18,100 --> 00:54:19,668
NON-CROSSOVERS, AND DOUBLE

1266
00:54:19,668 --> 00:54:22,504
HOLLIDAY JUNCTIONS WITHOUT

1267
00:54:22,504 --> 00:54:24,072
PRODUCING COSTOVERS.

1268
00:54:24,072 --> 00:54:25,240
SO NON-CROSSOVERS WERE DERIVED

1269
00:54:25,240 --> 00:54:27,776
FROM DOUBLE HOLLIDAY JUNCTION

1270
00:54:27,776 --> 00:54:28,777
BUS NON-CROSSOVER FORMATION WAS

1271
00:54:28,777 --> 00:54:32,681
INDEPENDENT OF THAT.

1272
00:54:32,681 --> 00:54:34,382
AND WHAT THORSTON SUGGESTED

1273
00:54:34,382 --> 00:54:41,756
THESE WERE FORMED BY DISTINCT

1274
00:54:41,756 --> 00:54:42,657
BIOCHEMICAL MECHANISMS,

1275
00:54:42,657 --> 00:54:45,527
CROSSOVER FORMED BY CLEAVAGE OF

1276
00:54:45,527 --> 00:54:46,495
HOLLIDAY JUNCTIONS BUT

1277
00:54:46,495 --> 00:54:48,763
NON-CROSSOVERS FORMED BY

1278
00:54:48,763 --> 00:54:50,198
COMPLETELY DIFFERENT PROCESS BY

1279
00:54:50,198 --> 00:54:54,236
WHICH THIS EARLY STRAND INVASION

1280
00:54:54,236 --> 00:54:57,005
INTERMEDIATE WAS TAKEN APART BY

1281
00:54:57,005 --> 00:55:04,880
HELICASE AND AKNEELED  -- 

1282
00:55:04,880 --> 00:55:10,352
A NEILSED, AFTER FILLING IN ONE

1283
00:55:10,352 --> 00:55:15,390
HAD A NON-CROSSOVER.

1284
00:55:15,390 --> 00:55:17,692
SINCE THEN, WE'VE LEARNED A LOT

1285
00:55:17,692 --> 00:55:18,026
MORE.

1286
00:55:18,026 --> 00:55:21,363
WE NOW KNOW THAT THIS PROCESS OF

1287
00:55:21,363 --> 00:55:23,064
CROSSOVER FORMATION FORMS IN

1288
00:55:23,064 --> 00:55:26,568
THE -- IS CARRIED OUT IN THE

1289
00:55:26,568 --> 00:55:29,504
CONTEXT OF AN ELABORATE PROTEIN

1290
00:55:29,504 --> 00:55:35,210
STRUCTURE THAT'S ASSEMBLED 

1291
00:55:35,210 --> 00:55:37,646
BETWEEN HOMOLOGOUS CHROMOSOMES,

1292
00:55:37,646 --> 00:55:40,949
SYNAPTIC COMPLEX, IN PARTICULAR

1293
00:55:40,949 --> 00:55:46,254
BY ZM PROTEINS, THESE IN EFFECT

1294
00:55:46,254 --> 00:55:49,424
BIND TO AND PROTECT

1295
00:55:49,424 --> 00:55:51,660
RECOMBINATION INTERMEDIATES FROM

1296
00:55:51,660 --> 00:55:52,394
DISASSEMBLY AND NON-CROSSOVER

1297
00:55:52,394 --> 00:55:59,067
FORMATION, AND SET THEM UP FOR 

1298
00:55:59,067 --> 00:56:02,337
THE RESOLUTION BY

1299
00:56:02,337 --> 00:56:03,538
MEIOSIS-SPECIFIC RESOLVEase TO

1300
00:56:03,538 --> 00:56:04,272
FORM CROSSOVERS.

1301
00:56:04,272 --> 00:56:06,274
THERE'S A THIRD PATHWAY OF

1302
00:56:06,274 --> 00:56:09,010
RECOMBINATION ACCOUNTING FOR A

1303
00:56:09,010 --> 00:56:11,046
MINOR FRACTION, 10% OF THE

1304
00:56:11,046 --> 00:56:13,315
PRODUCTS, WHICH OCCURS VIA

1305
00:56:13,315 --> 00:56:14,816
FORMATION OF INTERMEDIATE

1306
00:56:14,816 --> 00:56:17,018
SIMILAR TO THESE, ALTHOUGH THEY

1307
00:56:17,018 --> 00:56:19,221
HAVE MANY ALTERNATE STRUCTURES

1308
00:56:19,221 --> 00:56:27,028
AND THESE ARE, AGAIN, RESOLVED

1309
00:56:27,028 --> 00:56:31,566
UNDER CONTROL OF PROTEIN BUT A

1310
00:56:31,566 --> 00:56:32,968
SAID OF ENDONUCLEASES THAT ARE

1311
00:56:32,968 --> 00:56:39,374
NOT MEIOSIS SPECIFIC. 

1312
00:56:39,374 --> 00:56:41,443
THESE ARE RESOLVED TO FORM

1313
00:56:41,443 --> 00:56:44,613
CROSSOVERS AND NON-CROSSOVERS.

1314
00:56:44,613 --> 00:56:47,349
SO THE QUESTION THAT AROSE THEN,

1315
00:56:47,349 --> 00:56:50,218
WELL, IF THERE ARE THESE

1316
00:56:50,218 --> 00:56:52,954
MULTIPLE PATHWAYS FOR

1317
00:56:52,954 --> 00:56:54,222
RECOMBINATION, WHAT'S REGULATING

1318
00:56:54,222 --> 00:56:55,257
THE BRANCH POINT, WHAT'S

1319
00:56:55,257 --> 00:56:56,091
DETERMINING WHICH PATHWAY WILL

1320
00:56:56,091 --> 00:56:56,791
BE FOLLOWED?

1321
00:56:56,791 --> 00:57:05,333
AND THE ANSWER TO THAT IS THIS 

1322
00:57:05,333 --> 00:57:06,134
HELICASE, CONSERVED HELLICASE

1323
00:57:06,134 --> 00:57:10,038
COMPLEX THAT I'LL REFER TO AS

1324
00:57:10,038 --> 00:57:10,438
SDR.

1325
00:57:10,438 --> 00:57:12,307
WHAT I'LL TELL YOU IS NOW THE

1326
00:57:12,307 --> 00:57:17,579
LITTLE BIT OF EVIDENCE FOR THAT.

1327
00:57:17,579 --> 00:57:24,452
SO, STR IS THE HOMOLOG OF BTR, A

1328
00:57:24,452 --> 00:57:26,488
FAMILY, AND THESE GUYS ARE VERY

1329
00:57:26,488 --> 00:57:29,691
SIMILAR TO EACH OTHER BOTH IN

1330
00:57:29,691 --> 00:57:30,725
PRIMARY SEQUENCE AND STRUCTURE

1331
00:57:30,725 --> 00:57:34,296
BUT ALSO IN THE PHENOTYPES OF

1332
00:57:34,296 --> 00:57:37,666
MUTANTS THAT ONE SEES IN MUTANTS

1333
00:57:37,666 --> 00:57:38,967
LACKING THEM.

1334
00:57:38,967 --> 00:57:41,369
SO THEY BOTH SHOW PREMATURE

1335
00:57:41,369 --> 00:57:47,075
AGING, YES, YEAST GET OLD AND

1336
00:57:47,075 --> 00:57:48,410
AGE PREMATURELY.

1337
00:57:48,410 --> 00:57:49,711
THEY SHOW GENOME INSTABILITY.

1338
00:57:49,711 --> 00:57:52,981
AND MOST RELEVANT TO THIS TALK

1339
00:57:52,981 --> 00:57:54,516
INCREASED CROSSOVERS.

1340
00:57:54,516 --> 00:58:01,323
BOTH-- THIS IS AN ILLUSTRATION

1341
00:58:01,323 --> 00:58:06,127
FROM A VERY EARLY PIONEERING

1342
00:58:06,127 --> 00:58:07,896
PAPER SHOWING INCREASE THE

1343
00:58:07,896 --> 00:58:09,331
FREQUENCY OF SISTER CHROMATID

1344
00:58:09,331 --> 00:58:10,699
EXCHANGE IN THESE MUTANTS, EVERY

1345
00:58:10,699 --> 00:58:15,136
TIME YOU SEE A SWITCH FROM WHITE

1346
00:58:15,136 --> 00:58:18,807
OR GRAY TO BLACK IS A SISTER

1347
00:58:18,807 --> 00:58:19,908
CHROMATID RECOMBINATION.

1348
00:58:19,908 --> 00:58:23,945
OF COURSE, THESE PATIENTS ARE

1349
00:58:23,945 --> 00:58:25,613
ALSO HAVE PREDISPOSITION TO

1350
00:58:25,613 --> 00:58:26,047
CANCER.

1351
00:58:26,047 --> 00:58:27,115
YEAST DOESN'T GET CANCER, BUT

1352
00:58:27,115 --> 00:58:29,084
THE HOPE IS THAT BY

1353
00:58:29,084 --> 00:58:30,518
UNDERSTANDING WHAT'S GOING ON

1354
00:58:30,518 --> 00:58:34,255
HERE, WE CAN UNDERSTAND WHAT'S

1355
00:58:34,255 --> 00:58:36,758
GOING ON HERE.

1356
00:58:36,758 --> 00:58:37,125
OKAY.

1357
00:58:37,125 --> 00:58:40,362
OH, AND SO WHY IS THIS AN

1358
00:58:40,362 --> 00:58:43,098
ANTI-CROSSOVER, PREVENTING

1359
00:58:43,098 --> 00:58:44,165
CROSSOVERS?

1360
00:58:44,165 --> 00:58:47,068
THE HYPOTHESIS WAS THAT BTR OR

1361
00:58:47,068 --> 00:58:50,805
STR IS THE HELICASE THAT IS

1362
00:58:50,805 --> 00:58:51,539
INVOLVED IN NON-CROSSOVER

1363
00:58:51,539 --> 00:58:54,509
FORMATION, THAT IS IT'S TAKING

1364
00:58:54,509 --> 00:58:55,577
INTERMEDIATES THAT WOULD HAVE

1365
00:58:55,577 --> 00:58:56,811
MADE DOUBLE HOLLIDAY JUNCTIONS

1366
00:58:56,811 --> 00:58:58,813
AND THEN CROSSOVERS AND TAKEN

1367
00:58:58,813 --> 00:59:04,185
THEM APART AND TURNED THEM INTO

1368
00:59:04,185 --> 00:59:04,552
NON-CROSSOVERS.

1369
00:59:04,552 --> 00:59:06,020
THE IDEA WAS TO TEST THIS

1370
00:59:06,020 --> 00:59:10,492
HYPOTHESIS AND LOOK AT WHAT

1371
00:59:10,492 --> 00:59:14,329
HAPPENED DURING MEIOSIS IN SGS

1372
00:59:14,329 --> 00:59:20,668
1, TOP 3, RMI MUTANT. 

1373
00:59:20,668 --> 00:59:23,304
THE PREDICTION WE MADE, THE

1374
00:59:23,304 --> 00:59:24,372
THREE DID VARIOUS ASPECTS,

1375
00:59:24,372 --> 00:59:25,940
PREDICTION WAS IF IT WAS TAKING

1376
00:59:25,940 --> 00:59:30,512
THINGS APART TO MAKE

1377
00:59:30,512 --> 00:59:31,546
NON-CROSSOVERS IN AN SGS1 MUTANT

1378
00:59:31,546 --> 00:59:34,282
SHOULD SEE A LOSS OF

1379
00:59:34,282 --> 00:59:36,785
NON-CROSSOVERS AND POSSIBLY

1380
00:59:36,785 --> 00:59:40,255
INCREASE IN THESE CROSSOVERS.

1381
00:59:40,255 --> 00:59:42,123
AND THAT TURNED OUT TO BE TRUE,

1382
00:59:42,123 --> 00:59:49,497
BUT ALSO NOT TO BE TRUE.

1383
00:59:49,497 --> 00:59:55,303
SO AS I ALLUDED TO EARLIER, WHEN

1384
00:59:55,303 --> 01:00:00,241
WE LOOKED AT -- WHEN ANAUD 

1385
01:00:00,241 --> 01:00:02,110
LOOKED AT DNA LEVEL, IN

1386
01:00:02,110 --> 01:00:03,311
PARTICULAR UNDER CIRCUMSTANCES

1387
01:00:03,311 --> 01:00:06,681
WHERE HE BLOCKED FURTHER

1388
01:00:06,681 --> 01:00:08,316
PROGRESSION THROUGH THIS PROCESS

1389
01:00:08,316 --> 01:00:11,486
BY HAVING A -- BY CELLS THAT

1390
01:00:11,486 --> 01:00:14,155
FAILED TO EXPRESS CDC5 AND

1391
01:00:14,155 --> 01:00:17,659
TURNED ON CDC5, IN THE ABSENCE

1392
01:00:17,659 --> 01:00:24,899
OF CDC 5 HE SAW DOUBLE HOLLIDAY

1393
01:00:24,899 --> 01:00:26,267
JUNCTION INTERMEDIATES

1394
01:00:26,267 --> 01:00:26,935
ACCUMULATED, CROSSOVERS PRESENT

1395
01:00:26,935 --> 01:00:28,670
AT MUCH LOWER LEVEL, GRAPHS ARE

1396
01:00:28,670 --> 01:00:29,904
SHOWN HERE.

1397
01:00:29,904 --> 01:00:35,376
WHEN HE TURNED ON CDC5, HE SAW

1398
01:00:35,376 --> 01:00:38,146
RESOLUTION OF THE DOUBLE

1399
01:00:38,146 --> 01:00:39,914
HOLLIDAY JUNCTIONS, AND INCREASE

1400
01:00:39,914 --> 01:00:41,916
IN CROSSOVERS AS ONE WOULD

1401
01:00:41,916 --> 01:00:43,918
EXPECT IF DOUBLE HOLLIDAY

1402
01:00:43,918 --> 01:00:45,420
JUNCTIONS ARE RESOLVED TO FORM

1403
01:00:45,420 --> 01:00:47,188
CROSSOVERS BUT NO NET INCREASE

1404
01:00:47,188 --> 01:00:50,458
IN CROSSOVERS, AGAIN IN WILDTYPE

1405
01:00:50,458 --> 01:00:51,860
CELLS.

1406
01:00:51,860 --> 01:00:54,829
CONSISTENT WITH THE PICTURE THAT

1407
01:00:54,829 --> 01:00:57,665
THORSTON MADE THAT INDICATED

1408
01:00:57,665 --> 01:00:59,100
NON-CROSSOVERS WERE NOT FORMED

1409
01:00:59,100 --> 01:01:02,637
BY DOUBLE HOLLIDAY JUNCTION

1410
01:01:02,637 --> 01:01:04,005
INTERMEDIATE RESOLUTION.

1411
01:01:04,005 --> 01:01:07,141
HOWEVER, WHEN -- OH, I SHOULD

1412
01:01:07,141 --> 01:01:07,942
SAY THAT GENETICALLY IT WAS

1413
01:01:07,942 --> 01:01:10,645
SHOWN THAT THIS WAS THE MEIOTIC

1414
01:01:10,645 --> 01:01:15,016
RESOLVEASE THAT WAS DOING THIS.

1415
01:01:15,016 --> 01:01:17,352
HOWEVER, WHEN ARNAUD LOOKED AT

1416
01:01:17,352 --> 01:01:18,620
SGS1 MUTANTS HE FOUND A

1417
01:01:18,620 --> 01:01:20,221
DIFFERENT PICTURE.

1418
01:01:20,221 --> 01:01:22,323
IN THE ABSENCE OF CDC5

1419
01:01:22,323 --> 01:01:24,492
EXPRESSION WE DIDN'T GET

1420
01:01:24,492 --> 01:01:25,426
CROSSOVERS.

1421
01:01:25,426 --> 01:01:26,594
SAME AS IN WILDTYPE.

1422
01:01:26,594 --> 01:01:31,533
BUT WE ALSO DIDN'T GET

1423
01:01:31,533 --> 01:01:32,133
NON-CROSSOVERS.

1424
01:01:32,133 --> 01:01:36,571
AND WHEN YOU INDUCE CDC5 AND

1425
01:01:36,571 --> 01:01:38,840
INDUCED RESOLUTION EVER JOINT

1426
01:01:38,840 --> 01:01:39,741
MOLECULES, DOUBLE HOLLIDAY

1427
01:01:39,741 --> 01:01:41,910
JUNCTIONS, WE GOT BOTH

1428
01:01:41,910 --> 01:01:44,212
CROSSOVERS AND NON-CROSSOVERS.

1429
01:01:44,212 --> 01:01:48,483
SO, INSTEAD OF WIPING OUT

1430
01:01:48,483 --> 01:01:49,551
NON-CROSSOVER FORMATION

1431
01:01:49,551 --> 01:01:53,021
SPECIFICALLY, BOTH MEIOSIS --

1432
01:01:53,021 --> 01:01:53,922
OH, BOTH MEIOSIS-SPECIFIC

1433
01:01:53,922 --> 01:01:58,226
CROSSOVER FORMATION BUT ALSO

1434
01:01:58,226 --> 01:01:59,661
MEIOSIS-SPECIFIC NON-CROSSOVER

1435
01:01:59,661 --> 01:02:00,929
FORMATION HAD BEEN ABROGATED.

1436
01:02:00,929 --> 01:02:06,501
I SHOULD SAY THIS RESOLUTION WAS

1437
01:02:06,501 --> 01:02:11,673
NO LONGER DEPENDENT ON THE

1438
01:02:11,673 --> 01:02:14,409
MEIOTIC RESOLVEASE BUT WAS

1439
01:02:14,409 --> 01:02:15,176
DEPENDENT ON MITOTIC

1440
01:02:15,176 --> 01:02:15,476
RESOLVEASES.

1441
01:02:15,476 --> 01:02:20,048
THE LOSS OF SGS IS1 AND ALSO

1442
01:02:20,048 --> 01:02:23,251
TOP3 AND RMI1 RESULTED NOT ONLY

1443
01:02:23,251 --> 01:02:31,693
IN DEPLETION OF NON-CROSSOVER

1444
01:02:31,693 --> 01:02:32,293
PATHWAY BUT MEIOSIS-SPECIFIC

1445
01:02:32,293 --> 01:02:33,761
PATHWAY, A POPULATION OF THE

1446
01:02:33,761 --> 01:02:36,731
MINOR PATHWAY NOW ACCOUNTS FOR

1447
01:02:36,731 --> 01:02:38,700
ALL EVENTS WHEREBY JOINT

1448
01:02:38,700 --> 01:02:41,436
MOLECULES ARE FORMED AND THEY

1449
01:02:41,436 --> 01:02:44,272
ARE RESOLVED BY MITOTIC

1450
01:02:44,272 --> 01:02:44,772
RESOLVEASES.

1451
01:02:44,772 --> 01:02:46,574
SO THIS RAISED THE QUANDARY

1452
01:02:46,574 --> 01:02:55,216
BECAUSE AS I SAID WE THOUGHT OF

1453
01:02:55,216 --> 01:02:58,953
STR AS ENZYME THAT DISASSEMBLED

1454
01:02:58,953 --> 01:03:00,588
JOINT MOLECULES BUT DATA SAID IT

1455
01:03:00,588 --> 01:03:03,291
WAS MORE THAN FOR MAKING

1456
01:03:03,291 --> 01:03:04,726
NON-CROSSOVERS, THAT'S OKAY, BUT

1457
01:03:04,726 --> 01:03:08,329
ALSO IMPORTANT FOR THE MEIOTIC

1458
01:03:08,329 --> 01:03:10,331
CROSSOVER PATHWAY WHICH IS

1459
01:03:10,331 --> 01:03:12,166
BASICALLY ALL ABOUT PROTECTING

1460
01:03:12,166 --> 01:03:13,801
AND PRESERVING AND MAINTAINING

1461
01:03:13,801 --> 01:03:14,602
JOINT MOLECULES.

1462
01:03:14,602 --> 01:03:17,038
SO HOW COULD SOMETHING THAT

1463
01:03:17,038 --> 01:03:21,042
TAKES APART JOINT MOLECULES BE 

1464
01:03:21,042 --> 01:03:23,911
INVOLVED IN A PATHWAY THAT'S

1465
01:03:23,911 --> 01:03:25,346
ABOUT PRESERVING THEM?

1466
01:03:25,346 --> 01:03:28,483
AND THE HYPOTHESIS THAT WE CAME

1467
01:03:28,483 --> 01:03:38,993
UP WITH WAS THAT IN FACT  STR

1468
01:03:39,193 --> 01:03:40,895
WAS A CHAPERONE, YOU THINK ABOUT

1469
01:03:40,895 --> 01:03:43,231
PROMOTING A PROCESS LIKE FOLDING

1470
01:03:43,231 --> 01:03:44,732
PROTEIN BUT MOST CHAPERONES ARE

1471
01:03:44,732 --> 01:03:47,502
NOT THINGS THAT DRIVE FOLDING,

1472
01:03:47,502 --> 01:03:48,970
THEY ARE THINGS THAT UNFOLD

1473
01:03:48,970 --> 01:03:51,005
THINGS OR THAT TAKE THINGS

1474
01:03:51,005 --> 01:03:51,239
APART.

1475
01:03:51,239 --> 01:03:54,776
AND BY TAKING THINGS APART, WHAT

1476
01:03:54,776 --> 01:03:56,811
THEY DO IS THEY TAKE THINGS THAT

1477
01:03:56,811 --> 01:03:58,946
HAVE STARTED GOING THE WRONG

1478
01:03:58,946 --> 01:04:01,215
WAY, OKAY, AND RETURNED THEM TO

1479
01:04:01,215 --> 01:04:02,850
THE GROUND STATE, SO THEY CAN

1480
01:04:02,850 --> 01:04:03,951
TRY AGAIN.

1481
01:04:03,951 --> 01:04:08,589
AND THEY KEEP DOING THIS AND THE

1482
01:04:08,589 --> 01:04:12,827
FINAL STATE THAT THE PROPERLY

1483
01:04:12,827 --> 01:04:14,929
FOLDED STATE IS ONE A CHAPERONE

1484
01:04:14,929 --> 01:04:16,164
NO LONGER RECOGNIZED.

1485
01:04:16,164 --> 01:04:18,299
WE ARGUED, OKAY, THIS IS WHAT'S

1486
01:04:18,299 --> 01:04:20,401
HAPPENING HERE.

1487
01:04:20,401 --> 01:04:21,803
THESE INTERMEDIATES ARE

1488
01:04:21,803 --> 01:04:23,304
VULNERABLE TO STR AND GET TAKEN

1489
01:04:23,304 --> 01:04:24,972
APART AND THEY CAN BE RETURNED

1490
01:04:24,972 --> 01:04:27,375
BACK TO THE GROUND STATE, TO THE

1491
01:04:27,375 --> 01:04:29,143
DOUBLE STRAND BREAK STATE.

1492
01:04:29,143 --> 01:04:30,078
OKAY.

1493
01:04:30,078 --> 01:04:33,214
AND THIS WILL GO THROUGH

1494
01:04:33,214 --> 01:04:35,683
MULTIPLE CYCLES OF -- WE

1495
01:04:35,683 --> 01:04:37,819
HYPOTHESIZE MULTIPLE CYCLES OF

1496
01:04:37,819 --> 01:04:42,023
INVASION, DISASSEMBLY, NULL 

1497
01:04:42,023 --> 01:04:42,890
UNTIL EITHER NON-CROSSOVERS

1498
01:04:42,890 --> 01:04:44,959
WERE -- YOU FORMED A

1499
01:04:44,959 --> 01:04:46,494
NON-CROSSOVER WHICH CAN'T BE

1500
01:04:46,494 --> 01:04:52,233
RECOGNIZED BY STR OR THESE 

1501
01:04:52,233 --> 01:04:54,001
INTERMEDIATES WERE CAPTURED BY

1502
01:04:54,001 --> 01:04:58,406
THE ZMM PROTEINS OF THE

1503
01:04:58,406 --> 01:04:59,173
MEIOSIS-SPECIFIC STRUCTURE, THEN

1504
01:04:59,173 --> 01:05:01,409
ALLOWED TO MATURE, THEY WERE

1505
01:05:01,409 --> 01:05:02,143
ALSO PROTECTED.

1506
01:05:02,143 --> 01:05:02,443
OKAY.

1507
01:05:02,443 --> 01:05:05,880
SO ONE OF THE PREDICTIONS WAS

1508
01:05:05,880 --> 01:05:10,017
THAT IF WE WENT THROUGH MULTIPLE

1509
01:05:10,017 --> 01:05:15,990
CYCLES OF INVASION AND

1510
01:05:15,990 --> 01:05:19,193
DISASSEMBLY WE OUGHT TO SEE IT

1511
01:05:19,193 --> 01:05:21,829
AS TEMPLATE SWITCHING OR MOSAIC

1512
01:05:21,829 --> 01:05:23,397
HETERODUPLEX WITHIN THE PRODUCTS

1513
01:05:23,397 --> 01:05:24,465
OF RECOMBINATION.

1514
01:05:24,465 --> 01:05:26,501
BASICALLY I'VE DRAWN A

1515
01:05:26,501 --> 01:05:28,636
HYPOTHETICAL CASE WHERE THE BLUE

1516
01:05:28,636 --> 01:05:29,203
CHROMOSOME INVADES THE RED,

1517
01:05:29,203 --> 01:05:31,372
COPIES A LITTLE BIT OF RED, GETS

1518
01:05:31,372 --> 01:05:32,774
TAKEN APART, NOW IT HAS A LITTLE

1519
01:05:32,774 --> 01:05:34,275
BIT OF RED AGAIN.

1520
01:05:34,275 --> 01:05:37,378
HERE IT CAN NOW INVADE A BLUE

1521
01:05:37,378 --> 01:05:39,013
CHROMOSOME, COPY SOME BLUE, TAKE

1522
01:05:39,013 --> 01:05:40,314
APART AGAIN, ET CETERA, ET

1523
01:05:40,314 --> 01:05:41,115
CETERA.

1524
01:05:41,115 --> 01:05:44,919
AND IN THE END, WHAT ONE OBTAINS

1525
01:05:44,919 --> 01:05:46,521
ARE PRODUCTS OF RECOMBINATION

1526
01:05:46,521 --> 01:05:49,557
WHERE THERE ARE STRETCHES,

1527
01:05:49,557 --> 01:05:52,260
PATCHES OF RED DNA THAT ARE

1528
01:05:52,260 --> 01:05:53,828
INTERSPERSED WITH FULLY BLUE

1529
01:05:53,828 --> 01:05:54,061
DNA.

1530
01:05:54,061 --> 01:05:56,597
THE SAME IS TRUE OF CROSSOVERS.

1531
01:05:56,597 --> 01:05:59,534
AND SO THE PREDICTION WAS THAT

1532
01:05:59,534 --> 01:06:02,503
IF WE WENT THROUGH, IF THESE

1533
01:06:02,503 --> 01:06:09,143
MULTIPLE CYCLES OF INVASION AND

1534
01:06:09,143 --> 01:06:10,244
DISASSEMBLY WERE TRUE WE SHOULD

1535
01:06:10,244 --> 01:06:13,648
SEE EVIDENCE IN PRODUCTS OF

1536
01:06:13,648 --> 01:06:14,816
TEMPLATE SWITCHING.

1537
01:06:14,816 --> 01:06:16,717
A POSTDOC IN THE LAB SET OUT TO

1538
01:06:16,717 --> 01:06:17,685
DO THIS.

1539
01:06:17,685 --> 01:06:20,321
AND IN DOING SO, HE WENT BACK TO

1540
01:06:20,321 --> 01:06:24,458
THE VERY BEGINNING, TO TETRAD

1541
01:06:24,458 --> 01:06:28,095
DISSECTION AND ANALYSIS OF THE

1542
01:06:28,095 --> 01:06:33,334
DNA CONTENT OF THE  SPORES THAT

1543
01:06:33,334 --> 01:06:41,108
ARE PRODUCTS OF MEIOSIS, SET UP

1544
01:06:41,108 --> 01:06:44,278
A MODEL, A RECOMBINATION --

1545
01:06:44,278 --> 01:06:45,980
LOCUS TO STUDY RECOMBINATION,

1546
01:06:45,980 --> 01:06:48,649
ABLE TO INSERT MULTIPLE

1547
01:06:48,649 --> 01:06:50,084
POLYMORPHISMS AT REGULARLY

1548
01:06:50,084 --> 01:06:52,453
SPACED INTERVALS, WHICH HAD A

1549
01:06:52,453 --> 01:06:54,589
SINGLE DOUBLE-STRAND BREAK IN IT

1550
01:06:54,589 --> 01:06:56,724
SO WE KNEW WHERE RECOMBINATION

1551
01:06:56,724 --> 01:06:58,492
INITIATED AND WE COULD THEN

1552
01:06:58,492 --> 01:07:00,161
FOLLOW AMONG THE PRODUCTS OF

1553
01:07:00,161 --> 01:07:01,896
RECOMBINATION WHETHER IT HAD A

1554
01:07:01,896 --> 01:07:03,464
STRAND FROM THIS PARENT OR

1555
01:07:03,464 --> 01:07:06,133
STRAND FROM THAT PARENT.

1556
01:07:06,133 --> 01:07:08,903
SIMPLY BY SEQUENCING DNA

1557
01:07:08,903 --> 01:07:10,738
EXTRACTED FROM COLONIES FROM

1558
01:07:10,738 --> 01:07:11,739
THESE SPORES.

1559
01:07:11,739 --> 01:07:19,413
SO THIS IS ACTUALLY A MASSIVE 

1560
01:07:19,413 --> 01:07:19,947
UNDERTAKING.

1561
01:07:19,947 --> 01:07:23,217
IT INVOLVED DOING PACK BIOSIS

1562
01:07:23,217 --> 01:07:24,852
SEQUENCING, MULTIPLE MILLIONS OF

1563
01:07:24,852 --> 01:07:27,755
READS, AND THEN SORTING OUT

1564
01:07:27,755 --> 01:07:34,662
THOSE READS AND REASSEMBLING

1565
01:07:34,662 --> 01:07:40,001
THEM INTO THIS. 

1566
01:07:40,001 --> 01:07:43,571
HE WOULD GET AN E-MAIL FROM THE

1567
01:07:43,571 --> 01:07:44,205
BIOWULF FOLKS BECAUSE THEY

1568
01:07:44,205 --> 01:07:45,806
COULDN'T FIGURE OUT WHAT COULD

1569
01:07:45,806 --> 01:07:48,109
BE MAKING SUCH HEAVY USE OF THE

1570
01:07:48,109 --> 01:07:50,912
CORES HE HAD REQUESTED.

1571
01:07:50,912 --> 01:07:54,215
BUT NEVERTHELESS, HE DID DO IT.

1572
01:07:54,215 --> 01:07:57,251
AND THE ANSWER WAS VERY CLEAR.

1573
01:07:57,251 --> 01:07:59,020
THAT BOTH THE NON-CROSSOVER AND

1574
01:07:59,020 --> 01:08:02,390
THE CROSSOVER PRODUCTS OF

1575
01:08:02,390 --> 01:08:03,391
MEIOTIC RECOMBINATION SHOWED

1576
01:08:03,391 --> 01:08:05,192
EVIDENCE, FREQUENT EVIDENCE --

1577
01:08:05,192 --> 01:08:06,227
EVIDENCE OF FREQUENT TEMPLATE

1578
01:08:06,227 --> 01:08:06,994
SWITCHING WHERE, YOU KNOW,

1579
01:08:06,994 --> 01:08:09,730
THINGS WENT FROM RED TO BLUE TO

1580
01:08:09,730 --> 01:08:12,633
RED TO BLUE STRAND, ET CETERA,

1581
01:08:12,633 --> 01:08:17,672
ET CETERA.

1582
01:08:17,672 --> 01:08:22,109
SO, THE ANSWER TO THE QUESTION

1583
01:08:22,109 --> 01:08:23,644
IS, YES, MEIOTIC RECOMBINATION

1584
01:08:23,644 --> 01:08:26,414
INVOLVES MULTIPLE CYCLES OF

1585
01:08:26,414 --> 01:08:27,915
ASSEMBLY AND DISASSEMBLY.

1586
01:08:27,915 --> 01:08:29,450
SO, THAT'S BASICALLY THE STORY I

1587
01:08:29,450 --> 01:08:37,191
WANTED TO TELL YOU TODAY.

1588
01:08:37,191 --> 01:08:39,827
JUST TO SUMMARIZE THEN, THAT

1589
01:08:39,827 --> 01:08:43,197
MEIOTIC RECOMBINATION AND BY

1590
01:08:43,197 --> 01:08:45,833
INFERENCE WE THINK REPAIR OF

1591
01:08:45,833 --> 01:08:48,369
DOUBLE-STRAND BREAKS IN MITOTIC

1592
01:08:48,369 --> 01:08:50,671
CELLS IS A REMARKABLY DYNAMIC

1593
01:08:50,671 --> 01:08:52,306
PROCESS, IT INVOLVES MULTIPLE

1594
01:08:52,306 --> 01:08:55,943
CYCLES OF INVASION AND

1595
01:08:55,943 --> 01:08:57,545
DISASSOCIATION, IT INVOLVES

1596
01:08:57,545 --> 01:08:58,446
FREQUENT TEMPLATE SWITCHING,

1597
01:08:58,446 --> 01:08:59,747
SWITCHING FROM ONE REPAIR

1598
01:08:59,747 --> 01:09:05,353
TEMPLATE TO THE OTHER.

1599
01:09:05,353 --> 01:09:08,222
AND THAT IS DRIVEN BY HELICASES.

1600
01:09:08,222 --> 01:09:13,194
WHAT I HAVEN'T TOLD YOU TODAY IS

1601
01:09:13,194 --> 01:09:18,699
THAT SIMILARLY, THE OTHER STEPS,

1602
01:09:18,699 --> 01:09:20,935
SECOND STEP OF RECOMBINATION,

1603
01:09:20,935 --> 01:09:23,637
FORMATION OF DOUBLE HOLLIDAY

1604
01:09:23,637 --> 01:09:26,440
JUNCTIONS IS SIMILARLY DYNAMIC,

1605
01:09:26,440 --> 01:09:27,341
THAT HE'S DOUBLE HOLLIDAY

1606
01:09:27,341 --> 01:09:31,812
JUNCTIONS FREQUENTLY GO MOVEMENT

1607
01:09:31,812 --> 01:09:35,216
ACROSS THE CHROMOSOME, THAT

1608
01:09:35,216 --> 01:09:40,054
BRANCH MIGRATION ADDS ANOTHER

1609
01:09:40,054 --> 01:09:41,789
LAYER OF DYNAMISM, IF THEY

1610
01:09:41,789 --> 01:09:43,891
BRANCH MIGRATE IN ONE DIRECTION

1611
01:09:43,891 --> 01:09:45,926
WILL BECOME DISASSEMBLED AND

1612
01:09:45,926 --> 01:09:50,531
RETURNED BACK TO THIS STATE.

1613
01:09:50,531 --> 01:09:54,502
SO, AGAIN, THERE'S POTENTIAL

1614
01:09:54,502 --> 01:09:56,937
REVERSAL OF BOTH THE EARLIER AND

1615
01:09:56,937 --> 01:10:00,174
THE LATER STEPS OF MEIOTIC

1616
01:10:00,174 --> 01:10:00,508
RECOMBINATION.

1617
01:10:00,508 --> 01:10:02,209
SO, WHAT'S THE POINT HERE?

1618
01:10:02,209 --> 01:10:03,677
WHY GO DO ALL THIS STUFF IF YOU

1619
01:10:03,677 --> 01:10:07,915
JUST ARE GOING TO BACK OUT

1620
01:10:07,915 --> 01:10:09,450
AGAIN?

1621
01:10:09,450 --> 01:10:12,620
AND I THINK THERE ARE TWO -- AT

1622
01:10:12,620 --> 01:10:15,456
LEAST TWO POTENTIAL REASONS FOR

1623
01:10:15,456 --> 01:10:15,990
THIS.

1624
01:10:15,990 --> 01:10:22,229
THE FIRST IS -- INVOLVES THE

1625
01:10:22,229 --> 01:10:24,799
FACT THAT HOMOLOG PAIRING, THE

1626
01:10:24,799 --> 01:10:30,971
PAIRING OF HOMOLOGOUS 

1627
01:10:30,971 --> 01:10:34,375
CHROMOSOMES DURING MEIOSIS,

1628
01:10:34,375 --> 01:10:37,311
INVOLVES THE INITIATION AND

1629
01:10:37,311 --> 01:10:40,281
INVASION AT MULTIPLE SITES ALONG

1630
01:10:40,281 --> 01:10:41,482
THESE CHROMOSOMES, MULTIPLE

1631
01:10:41,482 --> 01:10:43,717
TRANSIENT EVENTS ARE USED TO

1632
01:10:43,717 --> 01:10:48,789
DRIVE THE PAIRING OF HOMOLOGOUS

1633
01:10:48,789 --> 01:10:49,156
CHROMOSOMES.

1634
01:10:49,156 --> 01:10:51,158
IF ONE EVENT HAPPENS IN A

1635
01:10:51,158 --> 01:10:53,527
REPEATED SEQUENCE, WHICH HAS

1636
01:10:53,527 --> 01:10:55,729
POTENTIAL TO RECOMBINE WITH A

1637
01:10:55,729 --> 01:10:59,200
REPEATED SEQUENCE ON THE WRONG

1638
01:10:59,200 --> 01:11:02,103
CHROMOSOME, YOU CAN -- YOU END

1639
01:11:02,103 --> 01:11:04,038
UP WITH ECTOPIC INTERACTION, AND

1640
01:11:04,038 --> 01:11:06,373
THAT WILL INTERFERE WITH PAIRING

1641
01:11:06,373 --> 01:11:08,642
AND INTERFERE WITH SUBSEQUENT

1642
01:11:08,642 --> 01:11:13,781
CHROMOSOME SEGREGATION, SO THE 

1643
01:11:13,781 --> 01:11:16,417
ABILITY TO REVERSE INTERACTIONS

1644
01:11:16,417 --> 01:11:19,153
IS -- ALLOWS ONE TO THEN GO

1645
01:11:19,153 --> 01:11:21,288
ALONG THE PROPER PATHWAY OF

1646
01:11:21,288 --> 01:11:23,524
HOMOLOG PAIRING.

1647
01:11:23,524 --> 01:11:25,526
THE SECOND IS THAT HOMOLOG

1648
01:11:25,526 --> 01:11:27,695
PAIRING ITSELF BECAUSE IT'S

1649
01:11:27,695 --> 01:11:29,263
INVOLVING MULTIPLE INTERACTIONS

1650
01:11:29,263 --> 01:11:32,199
AT DIFFERENT PLACES, HAS THE

1651
01:11:32,199 --> 01:11:33,200
POTENTIAL FOR MAKING

1652
01:11:33,200 --> 01:11:34,768
ENTANGLEMENTS WHERE IN THIS CASE

1653
01:11:34,768 --> 01:11:36,370
TWO HOMOLOGS PAIRED HERE AND

1654
01:11:36,370 --> 01:11:38,339
HERE, HERE AND HERE, BUT THEY

1655
01:11:38,339 --> 01:11:40,074
ARE INTERDIGITATED AMONG EACH

1656
01:11:40,074 --> 01:11:44,411
OTHER, AND, AGAIN, THIS IS GOING

1657
01:11:44,411 --> 01:11:47,982
TO BE LETHAL FOR SUBSEQUENT

1658
01:11:47,982 --> 01:11:48,582
CHROMOSOME SEGREGATION AND SO

1659
01:11:48,582 --> 01:11:50,918
ONE WANTS TO BE ABLE TO BACK OUT

1660
01:11:50,918 --> 01:11:53,354
PERHAPS AT THIS LATER STAGE IN

1661
01:11:53,354 --> 01:11:56,290
ORDER TO, AGAIN, PAIR HOMOLOGS

1662
01:11:56,290 --> 01:11:56,624
PROPERLY.

1663
01:11:56,624 --> 01:11:59,093
AND THE LAST THING I WANT TO DO

1664
01:11:59,093 --> 01:12:01,295
IN THE MINUS 12 SECONDS THAT I

1665
01:12:01,295 --> 01:12:03,364
HAVE IS TO POINT OUT TO YOU

1666
01:12:03,364 --> 01:12:05,599
AGAIN AS I SAID EARLIER THAT

1667
01:12:05,599 --> 01:12:09,270
WHAT WE LEARN IN MEIOTIC -- IN

1668
01:12:09,270 --> 01:12:10,738
STUDY OF MEIOSIS IS ALMOST

1669
01:12:10,738 --> 01:12:13,274
CERTAINLY GOING TO BE APPLICABLE

1670
01:12:13,274 --> 01:12:16,944
IN THE STUDY OF DNA REPAIR UNDER

1671
01:12:16,944 --> 01:12:19,947
OTHER CIRCUMSTANCES.

1672
01:12:19,947 --> 01:12:22,383
IN FACT, REALLY WHAT MY ON

1673
01:12:22,383 --> 01:12:24,652
THETIC RECOMBINATION IS IS

1674
01:12:24,652 --> 01:12:27,354
MITOTIC RECOMBINATION BECAUSE A

1675
01:12:27,354 --> 01:12:30,157
BUNCH OF CHROMOSOME STRUCTURE.

1676
01:12:30,157 --> 01:12:33,160
THE PICTURE THAT WE'VE COME

1677
01:12:33,160 --> 01:12:35,229
ABOUT FOR MEIOTIC RECOMBINATION

1678
01:12:35,229 --> 01:12:38,866
IF ONE JUST TAKES OUT THE

1679
01:12:38,866 --> 01:12:40,167
MEIOTIC CHROMOSOME STRUCTURES

1680
01:12:40,167 --> 01:12:41,135
THERE'S A PRETTY ACCURATE

1681
01:12:41,135 --> 01:12:42,069
PICTURE OF WHAT'S HAPPENING IN

1682
01:12:42,069 --> 01:12:47,942
THE MY TO THE INCORPORATE

1683
01:12:47,942 --> 01:12:48,542
CELL -- MITOTIC CELL.

1684
01:12:48,542 --> 01:12:50,177
I'D LIKE TO THANK THE PEOPLE WHO

1685
01:12:50,177 --> 01:12:53,480
WORKED WITH ME OVER THE YEARS.

1686
01:12:53,480 --> 01:12:58,552
AND I WOULD BE REMISS IF I 

1687
01:12:58,552 --> 01:13:00,221
DIDN'T END THIS TALK WITH THE

1688
01:13:00,221 --> 01:13:03,724
WAY WE END OUR PAPERS WITH AN

1689
01:13:03,724 --> 01:13:06,493
ACKNOWLEDGMENT TO THE INTRAMURAL

1690
01:13:06,493 --> 01:13:08,696
RESEARCH PROGRAM, CENTER FOR

1691
01:13:08,696 --> 01:13:10,464
CANCER RESEARCH, NATIONAL CANCER

1692
01:13:10,464 --> 01:13:13,901
INSTITUTE, IN THIS CASE IN

1693
01:13:13,901 --> 01:13:16,604
PARTICULAR TO THE FOLKS AT CCR

1694
01:13:16,604 --> 01:13:20,574
IN GENOMICS CORE AND SEQUENCING 

1695
01:13:20,574 --> 01:13:21,976
FACILITY THAT MADE THIS POSSIBLE

1696
01:13:21,976 --> 01:13:24,778
AND THE FOLKS AT BIOWULF WHO

1697
01:13:24,778 --> 01:13:35,189
HELPED US CARRY OUT THE

1698
01:13:38,559 --> 01:13:39,927
ANALYSIS.

1699
01:13:39,927 --> 01:13:40,194
[APPLAUSE]

1700
01:13:40,194 --> 01:13:40,928
YEAH, I KNOW.

1701
01:13:40,928 --> 01:13:43,430
>> I'M VERY BAD AS A TIME

1702
01:13:43,430 --> 01:13:43,664
KEEPER.

1703
01:13:43,664 --> 01:13:46,267
>> IT WAS A MINUTE AND 12

1704
01:13:46,267 --> 01:13:51,605
SECONDS OVER. 

1705
01:13:51,605 --> 01:13:55,542
>> THANK YOU FOR A
WONDERFUL

1706
01:13:55,542 --> 01:13:56,410
TALK.

1707
01:13:56,410 --> 01:13:57,278
SPEAKING OF CONTINUITY, THEMES

1708
01:13:57,278 --> 01:14:02,616
AND SO FORTH, A LITTLE BIT MORE

1709
01:14:02,616 --> 01:14:04,885
ON DOUBLE-STRAND BREAKS AND YOUR

1710
01:14:04,885 --> 01:14:07,588
MEIOSIS TO MITOSIS I THINK WAS A

1711
01:14:07,588 --> 01:14:12,893
PERFECT SEGUE INTO THE NEXT 

1712
01:14:12,893 --> 01:14:14,728
SPEAKER, DR. ANDRE NUSSENZWEIG

1713
01:14:14,728 --> 01:14:17,231
IS NIH DISTINGUISHED

1714
01:14:17,231 --> 01:14:19,333
INVESTIGATOR ALSO AT NCI CCR,

1715
01:14:19,333 --> 01:14:25,205
AND HE WORKS ON HOW CELLS REPAIR

1716
01:14:25,205 --> 01:14:27,141
DNA, PARTICULARLY DOUBLE-STRAND

1717
01:14:27,141 --> 01:14:29,777
BREAKS, BUT IN THE CONTEXT OF

1718
01:14:29,777 --> 01:14:33,914
ANOMALOUS REPAIR, IF YOU WILL,

1719
01:14:33,914 --> 01:14:35,883
BY TRANSLOCATION AND INSERTION

1720
01:14:35,883 --> 01:14:38,385
OF PIECES OF DNA FROM ONE

1721
01:14:38,385 --> 01:14:41,221
CHROMOSOME INTO THE NEXT, AND

1722
01:14:41,221 --> 01:14:44,525
THE ROLE OF THOSE -- THAT

1723
01:14:44,525 --> 01:14:46,994
PROCESS AND ITS REGULATION IN

1724
01:14:46,994 --> 01:14:49,596
CANCER BIOLOGY. 

1725
01:14:49,596 --> 01:14:51,198
SO DR. NUSSENZWEIG.

1726
01:14:51,198 --> 01:15:00,507
[APPLAUSE] 

1727
01:15:00,507 --> 01:15:01,342
>> THANK YOU, NINA.

1728
01:15:01,342 --> 01:15:03,377
I WILL TALK MORE ABOUT DNA

1729
01:15:03,377 --> 01:15:05,846
BREAKS BUT WANTED TO EXPAND A

1730
01:15:05,846 --> 01:15:09,183
LITTLE BIT ABOUT MYSELF BECAUSE

1731
01:15:09,183 --> 01:15:13,053
I HAVE A GOTTEN HERE IN A

1732
01:15:13,053 --> 01:15:14,822
ROUNDABOUT WAY.

1733
01:15:14,822 --> 01:15:17,191
MY TRAINING WAS ORIGINALLY IN

1734
01:15:17,191 --> 01:15:17,424
PHYSICS.

1735
01:15:17,424 --> 01:15:19,693
LET ME SEE IF I CAN GET THIS

1736
01:15:19,693 --> 01:15:25,366
POINTER TO WORK. 

1737
01:15:25,366 --> 01:15:26,200
NOT SURE HOW.

1738
01:15:26,200 --> 01:15:28,068
WELL, SOMEONE CAN -- OH, THERE.

1739
01:15:28,068 --> 01:15:28,936
GOOD, GOOD.

1740
01:15:28,936 --> 01:15:31,105
MY TRAINING WAS IN PHYSICS.

1741
01:15:31,105 --> 01:15:35,609
I GOT A Ph.D. IN PHYSICS, THE

1742
01:15:35,609 --> 01:15:37,778
SAME DEPARTMENT AS MICHAEL

1743
01:15:37,778 --> 01:15:39,113
LICHTEN'S DADA PROFESSOR THERE

1744
01:15:39,113 --> 01:15:39,747
AT THE TIME.

1745
01:15:39,747 --> 01:15:45,219
I WENT ON AND DID A POSTDOC IN

1746
01:15:45,219 --> 01:15:47,187
PHYSICS AS WELL.

1747
01:15:47,187 --> 01:15:51,058
AND THEN, I STARTED TO DABBLE

1748
01:15:51,058 --> 01:15:57,631
INTO BIOLOGY, IN THE DEPARTMENT

1749
01:15:57,631 --> 01:16:01,435
OF MEDICAL PHYSICAL IT AT

1750
01:16:01,435 --> 01:16:01,802
SLOAN-KETTERING.

1751
01:16:01,802 --> 01:16:05,072
AL SINGER TOOK A RISK AND HIRED

1752
01:16:05,072 --> 01:16:08,342
ME IN EXPERIMENTAL IMMUNOLOGY

1753
01:16:08,342 --> 01:16:08,976
BRANCH.

1754
01:16:08,976 --> 01:16:11,812
AND THAT'S WHERE I LEARNED A LOT

1755
01:16:11,812 --> 01:16:13,480
ABOUT IMMUNOLOGY AND ALSO

1756
01:16:13,480 --> 01:16:16,617
STARTED WORKING ON DNA REPAIR.

1757
01:16:16,617 --> 01:16:21,054
AND I CONTINUED TO WORK ON DNA

1758
01:16:21,054 --> 01:16:23,023
REPAIR, AND I GUESS NCI LIKED

1759
01:16:23,023 --> 01:16:25,793
THE TOPIC OF DNA REPAIR, AND

1760
01:16:25,793 --> 01:16:29,396
THEY CREATED A BRANCH CALLED

1761
01:16:29,396 --> 01:16:30,330
LABORATORY OF GENOME INTEGRITY,

1762
01:16:30,330 --> 01:16:33,434
WHERE I CONTINUED TO WORK ON

1763
01:16:33,434 --> 01:16:38,572
THAT, BUT ALSO NOW FLIRTING WITH

1764
01:16:38,572 --> 01:16:39,406
NEUROBIOLOGY.

1765
01:16:39,406 --> 01:16:41,074
SO YOU MAY THINK THIS SOUNDS

1766
01:16:41,074 --> 01:16:49,383
LIKE THIS GUY IS A DILITANT, 

1767
01:16:49,383 --> 01:16:50,851
DABBLING IN DIFFERENT AREAS, BUT

1768
01:16:50,851 --> 01:16:53,020
I WANT TO DEFEND MYSELF BY

1769
01:16:53,020 --> 01:16:57,157
SAYING THE STUDY OF DNA REPAIR

1770
01:16:57,157 --> 01:16:59,860
ALLOWS YOU OR GIVES YOU MAJOR

1771
01:16:59,860 --> 01:17:01,528
INSIGHT INTO DIFFERENT AREAS,

1772
01:17:01,528 --> 01:17:05,466
I'VE BENEFITED AND LEARNED FROM 

1773
01:17:05,466 --> 01:17:06,867
THIS STUDY.

1774
01:17:06,867 --> 01:17:09,169
BECAUSE DNA DAMAGE OCCURS IN ALL

1775
01:17:09,169 --> 01:17:11,572
CELL TYPES, OCCURS IN MANY

1776
01:17:11,572 --> 01:17:12,673
DIFFERENT WAYS AS MICHAEL

1777
01:17:12,673 --> 01:17:18,378
LICHTEN POINTED OUT THROUGH

1778
01:17:18,378 --> 01:17:22,649
EXOGENOUS AGENTS LIKE CHEMICALS

1779
01:17:22,649 --> 01:17:24,551
IN THERAPEUTICS, AND WE'RE

1780
01:17:24,551 --> 01:17:25,118
INTERESTING IN ENDOGENOUS

1781
01:17:25,118 --> 01:17:30,424
SOURCES OF  DAMAGE, REOXYGEN

1782
01:17:30,424 --> 01:17:32,893
SPECIES THAT MIGHT OCCUR IN

1783
01:17:32,893 --> 01:17:34,661
NORMAL METABOLISM BY ENZYMES,

1784
01:17:34,661 --> 01:17:38,098
NUCLEASES THAT PURPOSELY CUT THE

1785
01:17:38,098 --> 01:17:42,336
DNA AS MICHAEL MENTIONED, RAG

1786
01:17:42,336 --> 01:17:47,774
AID, SPO11 AND THIS DEMETHase,

1787
01:17:47,774 --> 01:17:49,076
TET ENZYMES.

1788
01:17:49,076 --> 01:17:51,645
SO IF YOU STUDY THIS YOU CAN

1789
01:17:51,645 --> 01:17:53,080
LEARN ABOUT IMMUNOLOGY BECAUSE

1790
01:17:53,080 --> 01:17:55,249
THIS IS IMPORTANT FOR DIVERSITY

1791
01:17:55,249 --> 01:17:56,984
OF THE IMMUNE SYSTEM, RAG AND

1792
01:17:56,984 --> 01:18:00,053
AID, THIS IS IMPORTANT FOR THE

1793
01:18:00,053 --> 01:18:02,256
DIVERSITY OF THE SPECIES, SPO 11

1794
01:18:02,256 --> 01:18:05,292
AND TET ENZYMES ARE IMPORTANT 

1795
01:18:05,292 --> 01:18:09,563
FOR CELL IDENTITY.

1796
01:18:09,563 --> 01:18:10,664
OKAY.

1797
01:18:10,664 --> 01:18:13,934
AS MICHAEL POINTED OUT, THESE

1798
01:18:13,934 --> 01:18:15,135
ENDOGENOUS ENZYMES, NUCLEASES

1799
01:18:15,135 --> 01:18:16,570
ARE REALLY GOOD, REPAIR IS

1800
01:18:16,570 --> 01:18:18,272
REALLY GOOD BUT OCCASIONALLY YOU

1801
01:18:18,272 --> 01:18:20,507
HAVE MISTAKES AND THESE MISTAKES

1802
01:18:20,507 --> 01:18:23,277
HAVE CONSEQUENCES.

1803
01:18:23,277 --> 01:18:26,113
YOU GET MUTATIONS, CHROMOSOMAL

1804
01:18:26,113 --> 01:18:27,180
INSTABILITY, EPIGENETIC

1805
01:18:27,180 --> 01:18:28,282
ALTERATIONS.

1806
01:18:28,282 --> 01:18:31,718
AND THIS CAN LEAD TO TISSUE

1807
01:18:31,718 --> 01:18:35,956
DECLINE, AGING, AND OF COURSE

1808
01:18:35,956 --> 01:18:36,323
CANCER.

1809
01:18:36,323 --> 01:18:39,893
SO, YOU KNOW, BEING AT THE

1810
01:18:39,893 --> 01:18:41,862
CANCER INSTITUTE, THE FOCUS OF

1811
01:18:41,862 --> 01:18:45,165
MY LAB ANYWAYS IS ON BASIC

1812
01:18:45,165 --> 01:18:50,470
MECHANISMS OF CANCER, AND IT WAS

1813
01:18:50,470 --> 01:18:52,506
VERY INTERESTING THIS YEAR WHEN

1814
01:18:52,506 --> 01:18:56,877
THE NCI AND THE CANCER U.K.

1815
01:18:56,877 --> 01:18:59,279
ANNOUNCED THE CANCER GRAND

1816
01:18:59,279 --> 01:19:01,348
CHALLENGES, SO THESE WERE WHAT

1817
01:19:01,348 --> 01:19:04,084
THEY CONSIDERED TO BE NINE OF

1818
01:19:04,084 --> 01:19:05,886
THE TOUGHEST PROBLEMS, MOST

1819
01:19:05,886 --> 01:19:09,356
DIFFICULT PROBLEMS IN CANCER

1820
01:19:09,356 --> 01:19:11,191
RESEARCH, AND THESE NINE

1821
01:19:11,191 --> 01:19:12,693
PROBLEMS WERE BASICALLY A

1822
01:19:12,693 --> 01:19:15,228
CHALLENGE SO THAT YOU GET PEOPLE

1823
01:19:15,228 --> 01:19:16,930
FROM DIFFERENT AREAS TO

1824
01:19:16,930 --> 01:19:18,265
COLLABORATE, INTERACT, TRY TO

1825
01:19:18,265 --> 01:19:23,036
SOLVE THESE REALLY, REALLY TOUGH

1826
01:19:23,036 --> 01:19:23,437
PROBLEMS.

1827
01:19:23,437 --> 01:19:27,274
SO, WE STARTED WORKING ON THIS

1828
01:19:27,274 --> 01:19:28,475
PROBLEM, CHEMOTHERAPY INDUCED

1829
01:19:28,475 --> 01:19:29,109
NEUROTOXICITY.

1830
01:19:29,109 --> 01:19:30,978
WE DIDN'T JUST DECIDE TO WORK ON

1831
01:19:30,978 --> 01:19:31,411
IT.

1832
01:19:31,411 --> 01:19:32,980
LIKE EVERYTHING ELSE, YOU JUST

1833
01:19:32,980 --> 01:19:33,947
RUN INTO IT.

1834
01:19:33,947 --> 01:19:36,116
AND BUT WE BECAME VERY

1835
01:19:36,116 --> 01:19:36,550
INTERESTED IN THIS.

1836
01:19:36,550 --> 01:19:41,121
THIS IS -- YOU KNOW, REALLY A

1837
01:19:41,121 --> 01:19:42,155
CHALLENGING PROBLEM BECAUSE

1838
01:19:42,155 --> 01:19:45,792
CHEMOTHERAPY CONTINUES TO EXTEND

1839
01:19:45,792 --> 01:19:48,929
LIFESPAN OF PATIENTS, BUT THERE

1840
01:19:48,929 --> 01:19:51,231
IS FREQUENTLY PERSISTENT

1841
01:19:51,231 --> 01:19:53,200
NEUROLOGICAL SYSTEMS, SYMPTOMS,

1842
01:19:53,200 --> 01:19:55,168
AND COGNITIVE IMPAIRMENT WHICH

1843
01:19:55,168 --> 01:20:00,107
MAKES THE LIFE VERY, VERY

1844
01:20:00,107 --> 01:20:00,741
DIFFICULT.

1845
01:20:00,741 --> 01:20:02,042
AND THE CHALLENGE REALLY IS

1846
01:20:02,042 --> 01:20:04,711
BECAUSE THIS IS A SUPER

1847
01:20:04,711 --> 01:20:06,413
COMPLICATED PROBLEM.

1848
01:20:06,413 --> 01:20:08,582
THERE ARE MULTIPLE FACTORS THAT

1849
01:20:08,582 --> 01:20:12,986
CONTRIBUTE TO THIS COGNITIVE

1850
01:20:12,986 --> 01:20:13,587
IMPAIRMENT AFTER CHEMOTHERAPY,

1851
01:20:13,587 --> 01:20:16,523
INCLUDING AGE OF THE PATIENT,

1852
01:20:16,523 --> 01:20:18,558
GENDER, CANCER TYPE, DOSE,

1853
01:20:18,558 --> 01:20:22,195
CANCER REGIMEN, ET CETERA, ET

1854
01:20:22,195 --> 01:20:22,562
CETERA.

1855
01:20:22,562 --> 01:20:27,367
AND JUST TO GIVE AN EXAMPLE HOW

1856
01:20:27,367 --> 01:20:33,874
DIFFERENT CANCER REGIMENS CAUSE

1857
01:20:33,874 --> 01:20:36,943
NEUROLOGICAL CHANGES, WITH

1858
01:20:36,943 --> 01:20:42,749
TOXICITIES DEPENDING ON WHAT 

1859
01:20:42,749 --> 01:20:44,718
COMPOUND, FROM HEADACHE TO

1860
01:20:44,718 --> 01:20:55,262
MENTAL STATE, MEMORY, COGNITIVE

1861
01:20:56,196 --> 01:20:58,198
ABILITY, EVEN SEIZURE ONE

1862
01:20:58,198 --> 01:21:00,500
PARTICULAR TYPE OF CHEMOTHERAPY,

1863
01:21:00,500 --> 01:21:04,004
ANTI-METABOLITES, ARE SHOWN HERE

1864
01:21:04,004 --> 01:21:07,908
IN RED, WE BECAME INTERESTED IN

1865
01:21:07,908 --> 01:21:15,482
THEM, AND THEY INTERRUPT NUCLEIC

1866
01:21:15,482 --> 01:21:17,117
ACID SYNTHESIS.

1867
01:21:17,117 --> 01:21:18,518
IN PARTICULAR, WE WERE

1868
01:21:18,518 --> 01:21:23,190
INTERESTED IN STUDYING THESE SO

1869
01:21:23,190 --> 01:21:24,257
CALLED SUICIDE NUCLEOSIDES WHICH

1870
01:21:24,257 --> 01:21:25,192
TERMINATE DNA SYNTHESIS AND

1871
01:21:25,192 --> 01:21:30,130
THESE APPARENTLY ARE THE LARGEST

1872
01:21:30,130 --> 01:21:31,598
CLASS OF ANTI-NEOPLASTIC AGENTS,

1873
01:21:31,598 --> 01:21:40,707
20% OF CANCER THERAPIES USE

1874
01:21:40,707 --> 01:21:42,275
THESE SUICIDE ANTI-METABOLITES.

1875
01:21:42,275 --> 01:21:44,244
LET ME GO BACK.

1876
01:21:44,244 --> 01:21:46,446
THESE METABOLITES, WHAT THEY

1877
01:21:46,446 --> 01:21:49,216
BASICALLY ARE -- THEY MAINTAIN

1878
01:21:49,216 --> 01:21:50,016
NUCLEOBASE, PRETTY MUCH, BUT

1879
01:21:50,016 --> 01:21:54,755
THEY HAVE A SMALL CHANGE IN THE

1880
01:21:54,755 --> 01:21:57,924
SUGAR MOIETY, AND A RESULT OF

1881
01:21:57,924 --> 01:22:00,260
THIS CHANGE IS THAT THERE'S --

1882
01:22:00,260 --> 01:22:10,704
WHEN YOU INCORPORATE THESE

1883
01:22:13,039 --> 01:22:16,409
ANTI-METABOLITES, THEY STOP

1884
01:22:16,409 --> 01:22:18,812
SYNTHESIS.

1885
01:22:18,812 --> 01:22:21,982
THIS KILLS RAPIDLY DIVIDING

1886
01:22:21,982 --> 01:22:23,950
CANCER CELLS AND ARREST DURING

1887
01:22:23,950 --> 01:22:26,019
REPLICATION OR THIS CAN GENERATE

1888
01:22:26,019 --> 01:22:34,995
DNA BREAKS AND MITOTIC CELL

1889
01:22:34,995 --> 01:22:35,195
DEATH.

1890
01:22:35,195 --> 01:22:37,497
WHAT ABOUT POST MITOTIC CELLS,

1891
01:22:37,497 --> 01:22:38,064
NEURONS?

1892
01:22:38,064 --> 01:22:42,369
HOW WOULD A SUICIDE

1893
01:22:42,369 --> 01:22:44,171
ANTI-METABOLITE IN NON-DIVIDING

1894
01:22:44,171 --> 01:22:46,907
CELLS HAVE THIS BAD EFFECT IN

1895
01:22:46,907 --> 01:22:47,507
POST MITOTIC NEURONS?

1896
01:22:47,507 --> 01:22:49,376
AND THEY DO.

1897
01:22:49,376 --> 01:22:50,076
THEY DEFINITELY DO.

1898
01:22:50,076 --> 01:22:54,247
AND THIS IS JUST ONE CLASSIC

1899
01:22:54,247 --> 01:22:58,084
EXAMPLE OF AN AGENT, THE

1900
01:22:58,084 --> 01:23:00,787
MAINSTAY WHICH HAS BEEN THE

1901
01:23:00,787 --> 01:23:04,191
MAINSTAY FOR 30 YEARS FOR AML

1902
01:23:04,191 --> 01:23:10,564
TREATMENT, AND KNOWN AT LEAST AT

1903
01:23:10,564 --> 01:23:15,468
HIGH DOSES INDUCES MAJOR

1904
01:23:15,468 --> 01:23:16,369
CEREBELLAR TOXICITY.

1905
01:23:16,369 --> 01:23:18,471
AND THERE'S ONE PARTICULAR CELL

1906
01:23:18,471 --> 01:23:21,274
TYPE ACTUALLY THAT IS PRIMARILY

1907
01:23:21,274 --> 01:23:31,818
AFFECTED IN TH CEREBELLUM, THAT 

1908
01:23:34,921 --> 01:23:41,027
IS PURKINJE CELL FOR BASIC 

1909
01:23:41,027 --> 01:23:41,294
MOVEMENT.

1910
01:23:41,294 --> 01:23:47,367
WE STARTED LOOKING AT

1911
01:23:47,367 --> 01:23:55,475
CYTARABINE, TREATED IPCs WITH

1912
01:23:55,475 --> 01:23:59,846
CYTARIBINE, IT IS VERY TOXICS. 

1913
01:23:59,846 --> 01:24:00,914
WE LOOKED AT POST-MITOTIC

1914
01:24:00,914 --> 01:24:04,651
NEURONS DERIVED FROM THE SAME

1915
01:24:04,651 --> 01:24:05,685
iPSCs, TREATED WITH

1916
01:24:05,685 --> 01:24:07,487
CYTARIBINE, WE FOUND THAT THIS

1917
01:24:07,487 --> 01:24:11,091
WAS ALSO VERY TOXIC IN THE

1918
01:24:11,091 --> 01:24:11,858
NEURONS, NON-DIVIDING CELLS.

1919
01:24:11,858 --> 01:24:13,827
AND SO, YOU KNOW, THE CHALLENGE,

1920
01:24:13,827 --> 01:24:17,898
THE GRAND CHALLENGE HERE IS TO

1921
01:24:17,898 --> 01:24:19,299
UNDERSTAND AND PREVENT

1922
01:24:19,299 --> 01:24:20,934
CHEMOTHERAPY INDUCED

1923
01:24:20,934 --> 01:24:21,735
NEUROTOXICITY AND NEUROPATHY.

1924
01:24:21,735 --> 01:24:24,671
AND SO I'M GOING TO TELL YOU

1925
01:24:24,671 --> 01:24:30,777
TODAY ABOUT RECENT STUDY WHERE

1926
01:24:30,777 --> 01:24:32,779
WE'RE LOOKING AT THIS

1927
01:24:32,779 --> 01:24:34,781
CYTARIBINE, HOW IT TRIGGERS CELL

1928
01:24:34,781 --> 01:24:39,452
DEATH IN POST MITOTIC NEURONS.

1929
01:24:39,452 --> 01:24:41,187
WE STARTED WITH CRISPR

1930
01:24:41,187 --> 01:24:43,423
INTERFERENCE SCREEN IN NEURONS,

1931
01:24:43,423 --> 01:24:44,891
WE KNOW CYTARIBINE IS TOXIC,

1932
01:24:44,891 --> 01:24:47,093
WANTED TO SEE WHICH GENES IF WE

1933
01:24:47,093 --> 01:24:51,431
KNOCKED THEM OUT WOULD DECREASE

1934
01:24:51,431 --> 01:24:52,499
TOXICITY.

1935
01:24:52,499 --> 01:24:53,900
WE PERFORMED CRISPR INTERFERENCE

1936
01:24:53,900 --> 01:24:55,101
SCREEN IN COLLABORATION WITH

1937
01:24:55,101 --> 01:24:58,204
MICHAEL WARD AT NINDS ACTUALLY.

1938
01:24:58,204 --> 01:25:01,341
SO HE HAS THE iPSCs WITH

1939
01:25:01,341 --> 01:25:05,045
CRISPR INTERFERENCE MACHINERY WE

1940
01:25:05,045 --> 01:25:07,580
CAN INACTIVATE TRANSCRIPTION OF

1941
01:25:07,580 --> 01:25:13,153
VARIOUS GENES USING THE Cas9

1942
01:25:13,153 --> 01:25:14,454
FUSED TO KRAB DOMAIN, INFECT

1943
01:25:14,454 --> 01:25:17,857
WITH LIBRARY OF GUIDES AND

1944
01:25:17,857 --> 01:25:21,494
DIFFERENTIATE OUR iPSCs INTO

1945
01:25:21,494 --> 01:25:23,263
NEURONS, SIMPLY WITH SINGLE

1946
01:25:23,263 --> 01:25:24,664
TRANSCRIPTION WITH INDUCIBLE

1947
01:25:24,664 --> 01:25:25,832
TRANSCRIPTION FACTOR, WHEN YOU

1948
01:25:25,832 --> 01:25:27,133
GET NEURONS THAT HAVE ALL OF

1949
01:25:27,133 --> 01:25:31,137
THESE GUIDES THAT ARE INHIBITING

1950
01:25:31,137 --> 01:25:33,840
INDIVIDUAL TRANSCRIPTION, WE

1951
01:25:33,840 --> 01:25:37,677
TREAT OR NOT WITH CYTARIBINE AND

1952
01:25:37,677 --> 01:25:40,146
LOOK FOR GUIDES THAT PREVENT

1953
01:25:40,146 --> 01:25:42,482
CELL DEATH IN THESE NEURONS.

1954
01:25:42,482 --> 01:25:45,919
AND WE FOUND A LOT OF

1955
01:25:45,919 --> 01:25:46,886
INTERESTING HITS, THE CURVE IS

1956
01:25:46,886 --> 01:25:51,257
NOT SHOWING BUT THE TOP HITS ARE

1957
01:25:51,257 --> 01:25:53,626
SHOWING. 

1958
01:25:53,626 --> 01:25:56,296
SOMEHOW THAT'S THE MAC-TO-PC

1959
01:25:56,296 --> 01:25:56,496
THING.

1960
01:25:56,496 --> 01:26:00,700
WE DIDN'T EXPECT IS THIS,

1961
01:26:00,700 --> 01:26:09,142
FACTORS INVOLVED IN METHYLATION,

1962
01:26:09,142 --> 01:26:10,343
THIS DNA METHYLASE WHICH WORKS

1963
01:26:10,343 --> 01:26:13,079
WITH THIS PARTNER, AND THIS IS

1964
01:26:13,079 --> 01:26:22,622
INVOLVES IN THE REVERSE PROCESS 

1965
01:26:22,622 --> 01:26:24,791
OF DNA DEMETHYLATION.

1966
01:26:24,791 --> 01:26:28,728
AND WE ACTUALLY WENT BACK AND 

1967
01:26:28,728 --> 01:26:37,470
CLONED INDIVIDUAL GUIDES FOR 

1968
01:26:37,470 --> 01:26:38,371
TDG, TREATED WITH CYTARIBINE,

1969
01:26:38,371 --> 01:26:41,608
WHEN YOU LOSE THESE GENES YOU

1970
01:26:41,608 --> 01:26:43,576
GET RESISTANCE TO CYTARIBINE

1971
01:26:43,576 --> 01:26:48,014
PARTICULARLY WITH TDG.

1972
01:26:48,014 --> 01:26:50,717
HOW OR OTHER, THE ARA-C IS

1973
01:26:50,717 --> 01:26:53,253
BLOCKING THIS PROCESS OF

1974
01:26:53,253 --> 01:26:54,421
METHYLATION/DEMETHYLATION, AND

1975
01:26:54,421 --> 01:26:57,657
THE CHANGES OF METHYLATION OCCUR

1976
01:26:57,657 --> 01:27:01,828
AT REGULATORY ELEMENTS LIKE

1977
01:27:01,828 --> 01:27:02,195
ENHANCERS DURING

1978
01:27:02,195 --> 01:27:04,064
DIFFERENTIATION, AS WELL AS IN

1979
01:27:04,064 --> 01:27:04,964
RESPONSE TO STIMULI.

1980
01:27:04,964 --> 01:27:11,037
SO YOU IMAGINE THAT YOU HAVE --

1981
01:27:11,037 --> 01:27:16,643
THIS METHYLATION OCCURS AT CG

1982
01:27:16,643 --> 01:27:17,410
DINUCLEOTIDES, YOU HAVE

1983
01:27:17,410 --> 01:27:19,279
ENHANCER, CONVERTING TO NEURON,

1984
01:27:19,279 --> 01:27:21,247
THAT ENHANCER HAS TO OPEN UP,

1985
01:27:21,247 --> 01:27:28,822
THEN IT BECOMES DEMETHYLATED.

1986
01:27:28,822 --> 01:27:32,892
DMT1 IS RESPONSIBLE FOR 

1987
01:27:32,892 --> 01:27:36,396
METHYLATION OF THE CYTOSINE AND

1988
01:27:36,396 --> 01:27:38,264
TET ENZYMES RESPONSIBLE TO

1989
01:27:38,264 --> 01:27:41,634
REVERSE THIS PROCESS, NOT JUST

1990
01:27:41,634 --> 01:27:42,802
GOING BACK TO UNMETHYLATED,

1991
01:27:42,802 --> 01:27:49,209
THERE'S A SERIES OF OXIDATION

1992
01:27:49,209 --> 01:27:52,946
STEPS WITH METHYL GROUP

1993
01:27:52,946 --> 01:27:55,148
OXIDIZED, TURNING TO

1994
01:27:55,148 --> 01:27:56,749
HYDROXYMETHYL CYTOSINE AND

1995
01:27:56,749 --> 01:27:59,619
OXIDIZED TO FORMAL FOLLOWED BY

1996
01:27:59,619 --> 01:28:02,755
CARBOXYL CYTOSINE, STILL NOT

1997
01:28:02,755 --> 01:28:04,757
ENOUGH.

1998
01:28:04,757 --> 01:28:11,531
THERE'S THIS ENZYME TDG THAT 

1999
01:28:11,531 --> 01:28:12,832
REMOVES AND RECOGNIZES, EXCISES,

2000
01:28:12,832 --> 01:28:17,036
YOU HAVE TO REPAIR AND GET BACK

2001
01:28:17,036 --> 01:28:20,640
TO UNMETHYLATED CYTOSINE.

2002
01:28:20,640 --> 01:28:23,009
TO TEST HOW ARA-C MIGHT AFFECT

2003
01:28:23,009 --> 01:28:27,147
THIS PROCESS WE DEVELOPED HIGH

2004
01:28:27,147 --> 01:28:30,216
RESOLUTION METHODS TO DETECT

2005
01:28:30,216 --> 01:28:30,850
METHYLATION AND DEMETHYLATION.

2006
01:28:30,850 --> 01:28:35,588
SO, THIS IS WHAT I OUTLINED TO

2007
01:28:35,588 --> 01:28:38,558
YOU AT THE DNA LEVEL, TET

2008
01:28:38,558 --> 01:28:40,727
ENZYMES OXIDIZES METHYL GROUPS

2009
01:28:40,727 --> 01:28:42,128
TO CHANGE METHYLCYTOSINE

2010
01:28:42,128 --> 01:28:43,897
EVENTUALLY TO FORMAL CYTOSINE

2011
01:28:43,897 --> 01:28:45,999
AND CARBOXYL CYTOSINE.

2012
01:28:45,999 --> 01:28:49,569
THIS IS RECOGNIZED BY TDG WHICH

2013
01:28:49,569 --> 01:28:55,074
POPS IT OUT, EXCISES THE SPACE, 

2014
01:28:55,074 --> 01:29:04,584
AND ENZYME AND NUCLEASE MAKES A

2015
01:29:04,584 --> 01:29:09,189
KNICK, 

2016
01:29:09,189 --> 01:29:14,494
OR A MORE COMPLEX A LONGER PATCH

2017
01:29:14,494 --> 01:29:16,429
IS REPAIRED BY BASIC EXCISION

2018
01:29:16,429 --> 01:29:19,165
REPAIR NOT ONLY INVOLVING

2019
01:29:19,165 --> 01:29:22,368
CYTOSINE BUT A FEW MORE BASES

2020
01:29:22,368 --> 01:29:23,369
ARE SYNTHESIZED.

2021
01:29:23,369 --> 01:29:25,338
WE DEVELOPED WHO WAYS TO LOOK AT

2022
01:29:25,338 --> 01:29:29,142
THIS PROCESS.

2023
01:29:29,142 --> 01:29:32,312
ONE BY USING DIDEOXY CYTOSINE TO

2024
01:29:32,312 --> 01:29:34,080
BOX THIS SHORT PATCH BASE

2025
01:29:34,080 --> 01:29:37,383
EXCISION REPAIR AND THEN GOING

2026
01:29:37,383 --> 01:29:39,152
TO DETECT THIS BY THIS PROCESS

2027
01:29:39,152 --> 01:29:42,088
I'LL DESCRIBE, THIS THING WE

2028
01:29:42,088 --> 01:29:42,355
DEVELOPED.

2029
01:29:42,355 --> 01:29:45,191
THIS METHOD. 

2030
01:29:45,191 --> 01:29:47,227
WE ALSO DETECTED LONG PATCH BY

2031
01:29:47,227 --> 01:29:48,928
INCORPORATING EDU, WE WOULD HOPE

2032
01:29:48,928 --> 01:29:49,562
GETS INCORPORATED SOMEWHERE

2033
01:29:49,562 --> 01:29:54,133
ALONG THE LINE AFTER THE 

2034
01:29:54,133 --> 01:29:54,934
CYTOSINE IS INCORPORATED.

2035
01:29:54,934 --> 01:29:58,438
SO THIS IS A SEQUENCING APPROACH

2036
01:29:58,438 --> 01:29:59,706
THAT WE CALL

2037
01:29:59,706 --> 01:30:00,306
SYNTHESIS-ASSOCIATED REPAIR

2038
01:30:00,306 --> 01:30:05,578
SEQUENCING, WHERE WE SIMPLY TAKE

2039
01:30:05,578 --> 01:30:09,382
OUR POST MITOTIC NEURONS, ADD

2040
01:30:09,382 --> 01:30:15,154
EDU, ASK WHETHER THIS LONG PATCH

2041
01:30:15,154 --> 01:30:17,790
REPAIR IS HAPPENING, WE CAN

2042
01:30:17,790 --> 01:30:20,827
BIOATEN LATE AND SONICATE AND

2043
01:30:20,827 --> 01:30:25,932
PULL DOWN AND SEQUENCE AND IF

2044
01:30:25,932 --> 01:30:27,433
THERE ARE PEAKS, SPECIFIC SITES

2045
01:30:27,433 --> 01:30:30,603
IN THE GENOME, POST MITOTIC

2046
01:30:30,603 --> 01:30:31,971
CELLS UNDERGOING DNA REPAIR.

2047
01:30:31,971 --> 01:30:34,173
IF THERE ARE NO PEAKS IT'S

2048
01:30:34,173 --> 01:30:35,408
RANDOM OR DOESN'T HAPPEN.

2049
01:30:35,408 --> 01:30:45,852
I WILL GET BACK TO ARA-C. 

2050
01:30:47,687 --> 01:30:51,024
WE DETECTED 50,000 HOT SPOTS OF

2051
01:30:51,024 --> 01:30:52,892
DNA REPAIR IN THE GENOME,

2052
01:30:52,892 --> 01:30:54,560
ABSOLUTELY NOT RANDOM BUT

2053
01:30:54,560 --> 01:30:56,729
ASSOCIATED WITH ENHANCERS, JUST

2054
01:30:56,729 --> 01:31:00,366
ADD EDU TO POST MITOTIC NEURONS

2055
01:31:00,366 --> 01:31:06,873
AND YOU SEE THESE SAR Seq 

2056
01:31:06,873 --> 01:31:08,775
PEAKS CORRELATE WITH ENHANCERS.

2057
01:31:08,775 --> 01:31:14,914
AND THEY CO-LOCALIZE WITH THE

2058
01:31:14,914 --> 01:31:16,916
ENZYME, METHYL TRANSFERASE,

2059
01:31:16,916 --> 01:31:18,518
DIRECTLY BINDS TO THESE SITES,

2060
01:31:18,518 --> 01:31:20,687
NOT JUST ANY OPEN DNA, NOT

2061
01:31:20,687 --> 01:31:24,991
PROMOTERS OR GENE BODIES BUT

2062
01:31:24,991 --> 01:31:26,292
SPECIFICALLY NEURONAL ENHANCERS.

2063
01:31:26,292 --> 01:31:28,594
SO THIS IS A NICE METHOD,

2064
01:31:28,594 --> 01:31:30,697
SURPRISING RESULT BUT WE WANTED

2065
01:31:30,697 --> 01:31:34,067
TO KNOW EXACTLY WHERE WITHIN THE

2066
01:31:34,067 --> 01:31:37,270
ENHANCERS DO THESE ENDOGENOUS

2067
01:31:37,270 --> 01:31:39,305
DNA REPAIR INTERMEDIATES OCCUR.

2068
01:31:39,305 --> 01:31:49,682
AND SO WE USED A CHAIN

2069
01:31:51,284 --> 01:31:51,784
TERMINATOR, DIDEOXYCYTOSINE,

2070
01:31:51,784 --> 01:31:52,919
HAPPENING IN POST MITOTIC CELLS

2071
01:31:52,919 --> 01:31:55,655
AND USE THIS TO TERMINATE THE

2072
01:31:55,655 --> 01:32:00,593
CRANE  -- THE CHAIN.

2073
01:32:00,593 --> 01:32:03,930
IF IT TERMINATES YOU HAVE A GAP,

2074
01:32:03,930 --> 01:32:07,133
WE TRY TO DETECT THE GAP USING

2075
01:32:07,133 --> 01:32:09,035
S1 NUCLEASE TO CUT THAT AND

2076
01:32:09,035 --> 01:32:09,836
WE'LL GENERATE NOW A

2077
01:32:09,836 --> 01:32:11,404
DOUBLE-STRAND BREAK WHICH WE

2078
01:32:11,404 --> 01:32:13,906
KNOW HOW TO DETECT.

2079
01:32:13,906 --> 01:32:17,944
SO BASICALLY PROTOCOL IS JUST TO

2080
01:32:17,944 --> 01:32:24,450
TAKE YOUR NEURONS, TREAT WITH

2081
01:32:24,450 --> 01:32:26,486
DIOXYCYTOSINE, EMBED THE CELLS

2082
01:32:26,486 --> 01:32:36,896
IN A PLUG AND THEN USE  

2083
01:32:42,201 --> 01:32:45,204
RECOMBINANT, AND WITH END-Seq

2084
01:32:45,204 --> 01:32:45,905
ATTACHING HAIRPIN ADAPTER TO

2085
01:32:45,905 --> 01:32:48,341
LEFT AND RIGHT END OF THE BREAK

2086
01:32:48,341 --> 01:32:50,810
AND WE WOULD GET A READ ON THE

2087
01:32:50,810 --> 01:32:52,078
BOTTOM, CORRESPONDING TO LEFT

2088
01:32:52,078 --> 01:32:53,613
END, AND READS ON THE TOP

2089
01:32:53,613 --> 01:32:58,017
CORRESPONDING TO THE RIGHT END.

2090
01:32:58,017 --> 01:33:01,454
SO, HERE WE HAVE OUR SAR-Seq,

2091
01:33:01,454 --> 01:33:03,289
DNA REPAIR SYNTHESIS,

2092
01:33:03,289 --> 01:33:04,724
INCORPORATE EDU.

2093
01:33:04,724 --> 01:33:06,793
RESOLUTION OF ABOUT 500 BASE

2094
01:33:06,793 --> 01:33:08,327
PAIRS OR SO.

2095
01:33:08,327 --> 01:33:11,097
AND OUR END-Seq BY ITSELF

2096
01:33:11,097 --> 01:33:13,933
THERE'S NO DOUBLE-STRANDED

2097
01:33:13,933 --> 01:33:14,267
BREAKS.

2098
01:33:14,267 --> 01:33:16,202
TREAT WITH S1, SEE NO GAPS, BUT

2099
01:33:16,202 --> 01:33:19,505
WHEN WE USE OUR CHAIN TERMINATOR

2100
01:33:19,505 --> 01:33:22,508
TO HALT THIS PROCESS AND THEN S1

2101
01:33:22,508 --> 01:33:27,613
AND THEN END-Seq WE SEE PEAKS.

2102
01:33:27,613 --> 01:33:31,717
THESE PEAKS OVERLAP WITH

2103
01:33:31,717 --> 01:33:35,521
SAR-Seq, HIGH RESOLUTION,

2104
01:33:35,521 --> 01:33:38,224
SINGLE NUCLEOTIDE RESOLUTION,

2105
01:33:38,224 --> 01:33:39,659
CLUSTERS OF SINGLE-STRAND BREAKS

2106
01:33:39,659 --> 01:33:45,798
THAT OVERLAP WITH BLOBS OF 

2107
01:33:45,798 --> 01:33:47,767
SAR-Seq PEAKS, WHEN WE LOOK

2108
01:33:47,767 --> 01:33:51,404
BACK WE SEE FIERY HIGH LEVEL OF

2109
01:33:51,404 --> 01:33:52,939
CG CONTENT.

2110
01:33:52,939 --> 01:33:55,575
CG METHYLATION, SO IT'S SMELLING

2111
01:33:55,575 --> 01:34:04,584
LIKE DNA DEMETHYLATION THAT WE 

2112
01:34:04,584 --> 01:34:06,252
ARE DETECTING.

2113
01:34:06,252 --> 01:34:07,887
SO WE WANTED TO -- THIS IS

2114
01:34:07,887 --> 01:34:09,822
ENDOGENOUS SOURCE.

2115
01:34:09,822 --> 01:34:11,691
WE'RE NOT GOING ANYTHING TO THE

2116
01:34:11,691 --> 01:34:12,191
NEURONS.

2117
01:34:12,191 --> 01:34:18,364
AND SO WE WANTED TO KNOW IS THIS

2118
01:34:18,364 --> 01:34:21,534
REALLY DNA DEMETHYLATION?

2119
01:34:21,534 --> 01:34:27,707
WE STARTED EXAMINING THIS

2120
01:34:27,707 --> 01:34:29,742
GLYCOSYLASE, DEVELOPED DEGRON

2121
01:34:29,742 --> 01:34:33,579
SYSTEM TO ENACT RAPIDLY TDG WITH

2122
01:34:33,579 --> 01:34:35,581
DETAG SYSTEM AND FOUND THE

2123
01:34:35,581 --> 01:34:41,487
SINGLE STRAND BREAKS YOU SEE

2124
01:34:41,487 --> 01:34:43,055
WITHOUT DTAG WERE ELIMINATED

2125
01:34:43,055 --> 01:34:44,790
WHEN YOU ADD, THIS IS THE HEAT

2126
01:34:44,790 --> 01:34:45,224
MAP.

2127
01:34:45,224 --> 01:34:47,593
BOTTOM LINE IS THAT WE FOUND

2128
01:34:47,593 --> 01:34:50,596
WITHOUT DOING ANYTHING TO THE

2129
01:34:50,596 --> 01:34:54,066
NEURONS JUST ADDING EDU SITES OF

2130
01:34:54,066 --> 01:34:56,569
DNA SYNTHESIS ASSOCIATED WITH

2131
01:34:56,569 --> 01:34:58,271
ENHANCERS, NEURONAL ENHANCERS,

2132
01:34:58,271 --> 01:35:02,508
AND THESE ARE GENERATED BECAUSE

2133
01:35:02,508 --> 01:35:07,213
OF THIS ENZYME, TET, WHICH

2134
01:35:07,213 --> 01:35:08,881
BASICALLY CONVERTS THE

2135
01:35:08,881 --> 01:35:09,615
METHYLCYTOSINE TO UNMODIFIED

2136
01:35:09,615 --> 01:35:12,885
CYTOSINE BUT THROUGH A PROCESS

2137
01:35:12,885 --> 01:35:15,721
THAT INVOLVES DNA DAMAGE AND

2138
01:35:15,721 --> 01:35:18,157
WE'RE -- WE BASICALLY USED OUR

2139
01:35:18,157 --> 01:35:20,426
CHAIN TERMINATOR TO DETECT THAT

2140
01:35:20,426 --> 01:35:21,861
INTERMEDIATE.

2141
01:35:21,861 --> 01:35:23,429
OKAY?

2142
01:35:23,429 --> 01:35:26,799
AND SO, THIS IS IMPORTANT, IT

2143
01:35:26,799 --> 01:35:29,001
HAPPENS IN, WE SEE SHORT PATCH

2144
01:35:29,001 --> 01:35:31,170
BASE EXCISION REPAIR AND LONG

2145
01:35:31,170 --> 01:35:32,438
PATCH BASE EXCISION REPAIR, AND

2146
01:35:32,438 --> 01:35:34,707
THIS PROCESS IS IMPORTANT FOR

2147
01:35:34,707 --> 01:35:37,643
CELL IDENTITY BUT ALSO THIS DNA

2148
01:35:37,643 --> 01:35:42,381
SYNTHESIS COULD LEAD TO -- COULD

2149
01:35:42,381 --> 01:35:43,783
CAUSE MUTAGENESIS ACTUALLY.

2150
01:35:43,783 --> 01:35:46,218
AND CHRIS WALSH GROUP AND PETER

2151
01:35:46,218 --> 01:35:50,489
PARK PUBLISHED A PAPER PRETTY

2152
01:35:50,489 --> 01:35:57,630
RECENTLY IN WHICH THEY SEQUENCED

2153
01:35:57,630 --> 01:36:00,333
NEURONS FROM POSTMORTEM PATIENTS

2154
01:36:00,333 --> 01:36:03,169
AND THEY FOUND THAT HUMAN

2155
01:36:03,169 --> 01:36:04,604
NEURONS HAVE ACTUALLY ACCUMULATE

2156
01:36:04,604 --> 01:36:07,440
POINT MUTATIONS AT A RATE OF 20

2157
01:36:07,440 --> 01:36:11,777
MUTATIONS PER NEURON PER YEAR.

2158
01:36:11,777 --> 01:36:16,682
AND THEY ALSO ACCUMULATE

2159
01:36:16,682 --> 01:36:17,817
INSERTION-DELETIONS ABOUT THREE

2160
01:36:17,817 --> 01:36:20,786
PER NEURON PER YEAR.

2161
01:36:20,786 --> 01:36:24,156
AND THEY -- MUTATIONS WERE NOT

2162
01:36:24,156 --> 01:36:26,058
RANDOM AND THEY COMPARED SITES

2163
01:36:26,058 --> 01:36:29,395
OF MUTATIONS WITH OUR HOT SPOTS

2164
01:36:29,395 --> 01:36:32,932
AND FOUND THAT ENHANCER

2165
01:36:32,932 --> 01:36:38,204
ASSOCIATED HOT SPOTS WERE 

2166
01:36:38,204 --> 01:36:39,238
ENRICHED FOR SOMATIC INDELS

2167
01:36:39,238 --> 01:36:40,840
FOUND IN THE PATIENT.

2168
01:36:40,840 --> 01:36:51,317
THIS MAY GO WRONG LEADING TO

2169
01:36:52,418 --> 01:36:52,585
INDELS.

2170
01:36:52,585 --> 01:36:54,387
BACK TO ARA-C, WHAT WOULD HAPPEN

2171
01:36:54,387 --> 01:37:03,362
IF YOU INTERRUPT THIS PROCESS OF

2172
01:37:03,362 --> 01:37:04,063
ACTIVE DEMETHYLATION WITH

2173
01:37:04,063 --> 01:37:04,730
CYTARIBINE.

2174
01:37:04,730 --> 01:37:07,133
IF THIS IS HAPPENING IN NEURONS

2175
01:37:07,133 --> 01:37:09,268
AND WE TREAT AND IT INTERRUPTS

2176
01:37:09,268 --> 01:37:10,903
THIS SYNTHESIS, WHAT IS THE CELL

2177
01:37:10,903 --> 01:37:12,805
GOING -- IS THE CELL GOING TO

2178
01:37:12,805 --> 01:37:15,841
DETECT IT AND RESPOND TO THAT

2179
01:37:15,841 --> 01:37:17,977
FORM OF INCOMPLETE DNA SYNTHESIS

2180
01:37:17,977 --> 01:37:18,911
OR DNA DAMAGE?

2181
01:37:18,911 --> 01:37:21,614
WE HAVE A VERY NICE WAY TO LOOK

2182
01:37:21,614 --> 01:37:24,917
AT DNA DAMAGE, THANKS TO BILL

2183
01:37:24,917 --> 01:37:28,421
BONNER, HERE AT NCI IN THE SAME

2184
01:37:28,421 --> 01:37:33,893
BUILDING 37 WHO DISCOVERED THIS

2185
01:37:33,893 --> 01:37:35,761
REMARKABLE HISTONE RARANT, H2AX,

2186
01:37:35,761 --> 01:37:38,931
WE USED HIS ANTIBODY AFTER

2187
01:37:38,931 --> 01:37:42,968
TREATING NEURONS WITH ARA-C, YOU

2188
01:37:42,968 --> 01:37:47,606
SEE THESE FOCI OF GAMMA H TO AX

2189
01:37:47,606 --> 01:37:48,974
INDICATING THERE'S A DNA DAMAGE

2190
01:37:48,974 --> 01:37:50,810
RESPONSE WHEN YOU TREAT NEURONS

2191
01:37:50,810 --> 01:37:51,510
WITH ARA-C.

2192
01:37:51,510 --> 01:37:54,513
IF YOU TREAT THE NEURONS WITH

2193
01:37:54,513 --> 01:37:59,819
ARA-A, ALSO VERY TOXIC TO CANCER

2194
01:37:59,819 --> 01:38:01,587
CELLS, THERE'S NO GAMMA-H2AX BUT

2195
01:38:01,587 --> 01:38:04,557
THERE'S A PROFOUND RESPONSE WITH

2196
01:38:04,557 --> 01:38:05,458
ARA-C.

2197
01:38:05,458 --> 01:38:09,595
DOES THIS DAMAGE RESPONSE

2198
01:38:09,595 --> 01:38:15,801
REQUIRE TET TDG?  WE USED OUR

2199
01:38:15,801 --> 01:38:17,870
DTAG SYSTEM.

2200
01:38:17,870 --> 01:38:23,809
YOU GET THESE ROBUST GAM H2AX

2201
01:38:23,809 --> 01:38:24,643
FOCI, INACTIVATE, DOESN'T HAPPEN

2202
01:38:24,643 --> 01:38:27,146
BECAUSE YOU DON'T EXCISE THE

2203
01:38:27,146 --> 01:38:28,481
BASE, REINCORPORATE ACA-C,

2204
01:38:28,481 --> 01:38:30,216
DOESN'T GET THERE.

2205
01:38:30,216 --> 01:38:31,717
INDEED I SHOWED YOU THAT NEURONS

2206
01:38:31,717 --> 01:38:35,121
ARE VERY, VERY SENSITIVE TO

2207
01:38:35,121 --> 01:38:36,922
ARA-C, THEY DIE.

2208
01:38:36,922 --> 01:38:40,392
IF YOU INACTIVATE TDG THEY DON'T

2209
01:38:40,392 --> 01:38:44,363
DIE BECAUSE THEY DON'T

2210
01:38:44,363 --> 01:38:45,731
INCORPORATE THE ARA-C, THERE'S

2211
01:38:45,731 --> 01:38:50,202
NO CELLULAR DNA DAMAGE RESPONSE 

2212
01:38:50,202 --> 01:38:53,339
SO CELLS DON'T DIE.

2213
01:38:53,339 --> 01:38:57,543
INTERESTING FACTS, MOST COMMON

2214
01:38:57,543 --> 01:39:01,113
CHEMOTHERAPY TO INDUCE 

2215
01:39:01,113 --> 01:39:08,020
CEREBELLAR FUNCTION IS HIGH

2216
01:39:08,020 --> 01:39:09,321
DOSE, LOSS OF PURKINJE CELLS,

2217
01:39:09,321 --> 01:39:11,557
THEY HAVE AMONG THE HIGHEST

2218
01:39:11,557 --> 01:39:15,561
LEVELS OF TET PROTEINS, SO MAYBE

2219
01:39:15,561 --> 01:39:19,298
THIS PROCESS IS HAPPENING MORE

2220
01:39:19,298 --> 01:39:23,335
SPECIFICALLY IN PURKINJE CELLS 

2221
01:39:23,335 --> 01:39:24,837
MAYBE CORRUPTED BY ARA-C.

2222
01:39:24,837 --> 01:39:27,439
WE STARTED TO INVESTIGATE THIS.

2223
01:39:27,439 --> 01:39:28,674
IN MICE.

2224
01:39:28,674 --> 01:39:31,043
SO WE TREAT MICE WITH ARA-C ON

2225
01:39:31,043 --> 01:39:35,047
DAILY BASIS FOR SEVEN DAYS, DO

2226
01:39:35,047 --> 01:39:38,317
BRAIN TISSUE SECTIONS, AND STAIN

2227
01:39:38,317 --> 01:39:39,285
FOR OUR GAMMA-H2AX.

2228
01:39:39,285 --> 01:39:42,221
AND THIS IS WHAT THE

2229
01:39:42,221 --> 01:39:42,988
CEREBELLUM -- WHAT A SECTION

2230
01:39:42,988 --> 01:39:44,456
LOOKS LIKE.

2231
01:39:44,456 --> 01:39:46,192
THESE ARE THE PURKINJE CELLS, IN

2232
01:39:46,192 --> 01:39:49,795
GREEN THE BIG CELLS WITH BIG

2233
01:39:49,795 --> 01:39:55,534
NUCLEOAND CYTOPLASM, STAINED BY 

2234
01:39:55,534 --> 01:39:59,605
CAL BINDING, GAMMA-H2AX IS

2235
01:39:59,605 --> 01:39:59,805
CLEAN.

2236
01:39:59,805 --> 01:40:01,507
IF YOU SEAT FOR SEVEN DAYS YOU

2237
01:40:01,507 --> 01:40:06,512
SEE FOCI AND THEY REALLY ARE

2238
01:40:06,512 --> 01:40:07,780
VERY SPECIFICALLY FOUND IN THE

2239
01:40:07,780 --> 01:40:09,315
PURKINJE CELLS.

2240
01:40:09,315 --> 01:40:11,584
THAT INDICATES MAYBE TET TDG IS

2241
01:40:11,584 --> 01:40:13,752
JUST ACTIVE FOR SOME REASON IN

2242
01:40:13,752 --> 01:40:14,486
PURKINJE CELLS.

2243
01:40:14,486 --> 01:40:17,022
AND THIS IS AN ADULT MOUSE.

2244
01:40:17,022 --> 01:40:21,460
AND SO WE THEN HAVE MICE

2245
01:40:21,460 --> 01:40:22,795
DEFICIENT IN TDG, TREAT WITH

2246
01:40:22,795 --> 01:40:27,600
ARA-C OR MICE THAT ARE KIND

2247
01:40:27,600 --> 01:40:30,336
OF -- THESE ARE THE

2248
01:40:30,336 --> 01:40:31,237
TDG-DEFICIENT MICE, AND THESE

2249
01:40:31,237 --> 01:40:33,038
ARE THE WILDTYPE MICE.

2250
01:40:33,038 --> 01:40:36,508
YOU CAN SEE A LITTLE FOCI FOR

2251
01:40:36,508 --> 01:40:37,843
GAMMA-H2AX HERE IN THE PURKINJE

2252
01:40:37,843 --> 01:40:41,847
CELLS AND THEY ARE LARGELY

2253
01:40:41,847 --> 01:40:43,449
ABSENT IN THE TDG-DEFICIENT

2254
01:40:43,449 --> 01:40:48,520
CELLS THAT ARE TREATED WITH

2255
01:40:48,520 --> 01:40:49,355
ARA-C.

2256
01:40:49,355 --> 01:40:51,056
WE DID FLOW CYTOMETRY BY

2257
01:40:51,056 --> 01:40:52,024
ISOLATING PURKINJE CELLS AND

2258
01:40:52,024 --> 01:40:54,193
THEN WHEN YOU SEE WILDTYPE CELLS

2259
01:40:54,193 --> 01:40:57,930
TREATED WITH ARA-C YOU CAN SEE A

2260
01:40:57,930 --> 01:41:00,766
SHIFT, GAMMA-H2AX HAPPENING IN

2261
01:41:00,766 --> 01:41:02,534
THE PURKINJE CELLS.

2262
01:41:02,534 --> 01:41:03,903
IN THE TDG DEFICIENT WHEN YOU DO

2263
01:41:03,903 --> 01:41:08,440
THE SAME YOU DON'T SEE THIS

2264
01:41:08,440 --> 01:41:12,278
SHIFT SO THIS INCORPORATION AND

2265
01:41:12,278 --> 01:41:14,079
GAMMA-H2AX FORMATION IS

2266
01:41:14,079 --> 01:41:16,181
DEPENDENT ON TDG.

2267
01:41:16,181 --> 01:41:19,485
OUR HYPOTHESIS IS THAT THE ARA-C

2268
01:41:19,485 --> 01:41:20,686
NEUROTOXICITY IS CAUSED BY

2269
01:41:20,686 --> 01:41:23,322
INTERRUPTING A PROCESS THAT IS

2270
01:41:23,322 --> 01:41:25,591
ACTUALLY IMPORTANT FOR NEURONAL

2271
01:41:25,591 --> 01:41:28,193
CELL IDENTITY, NAMELY THIS

2272
01:41:28,193 --> 01:41:30,696
ACTIVE DNA DEMETHYLATION.

2273
01:41:30,696 --> 01:41:36,001
IT'S BEEN USED,  CORNERSTONE OF

2274
01:41:36,001 --> 01:41:38,504
DRUGS FOR TREATMENT OF AML,

2275
01:41:38,504 --> 01:41:40,172
SPECIFICALLY WITH LOSS OF

2276
01:41:40,172 --> 01:41:41,840
PURKINJE CELLS, ONE SUGGESTION

2277
01:41:41,840 --> 01:41:42,975
IS WHAT ABOUT -- I DON'T KNOW

2278
01:41:42,975 --> 01:41:45,277
WHY PEOPLE DIDN'T USE THIS BUT

2279
01:41:45,277 --> 01:41:46,745
WHAT ABOUT USING ARA-A INSTEAD?

2280
01:41:46,745 --> 01:41:48,714
WE THINK THAT COULD KILL CANCER

2281
01:41:48,714 --> 01:41:49,581
CELLS.

2282
01:41:49,581 --> 01:41:52,484
BUT IT WOULD BE LESS TOXIC

2283
01:41:52,484 --> 01:41:54,186
BECAUSE IT WOULD AVOID

2284
01:41:54,186 --> 01:42:00,092
DOUBLE-STRAND BREAKS AT NEURONAL

2285
01:42:00,092 --> 01:42:00,492
ENHANCERS.

2286
01:42:00,492 --> 01:42:03,996
I DON'T KNOW WHAT THE TIMING IS

2287
01:42:03,996 --> 01:42:06,498
HERE.

2288
01:42:06,498 --> 01:42:09,168
>>  YOU HAVE A COUPLE
MINUTES.

2289
01:42:09,168 --> 01:42:10,636
>> I HAVE TWO MINUTES.

2290
01:42:10,636 --> 01:42:13,872
I WANT TO SHOW YOU CAN CHANGE A

2291
01:42:13,872 --> 01:42:17,009
CELL STATE AND ALSO INDUCE THE

2292
01:42:17,009 --> 01:42:18,344
SIMILAR PHENOMENON BY CHANGING

2293
01:42:18,344 --> 01:42:19,945
THE CELL STATE.

2294
01:42:19,945 --> 01:42:22,514
WE'RE GOING TO TURN

2295
01:42:22,514 --> 01:42:24,016
TRANSCRIPTIONAL CHANGES INTO

2296
01:42:24,016 --> 01:42:25,551
DOUBLE-STRANDED BREAKS.

2297
01:42:25,551 --> 01:42:27,119
SO HERE COMPLETELY DIFFERENT

2298
01:42:27,119 --> 01:42:28,087
SYSTEM.

2299
01:42:28,087 --> 01:42:31,390
WE TAKE BONE MARROW DERIVED

2300
01:42:31,390 --> 01:42:33,092
MACROPHAGES, GROW THEM, AND IN A

2301
01:42:33,092 --> 01:42:39,264
DISH, AND THEN WE TREAT THE

2302
01:42:39,264 --> 01:42:40,299
MACROPHAGES WITH LPS, WE'LL MAKE

2303
01:42:40,299 --> 01:42:43,702
A MAIN CHANGE IN THE

2304
01:42:43,702 --> 01:42:45,104
TRANSCRIPTIONAL RESPONSE.

2305
01:42:45,104 --> 01:42:48,540
AND YOU CAN SEE THAT IF YOU LOOK

2306
01:42:48,540 --> 01:42:49,608
AT FOUR DIFFERENT ENHANCERS AND

2307
01:42:49,608 --> 01:42:53,679
LOOK FOR MARKS OF ENHANCERS, YOU

2308
01:42:53,679 --> 01:42:55,848
CAN SEE LIKE H3K27 ACETYLATION,

2309
01:42:55,848 --> 01:42:56,949
WITHOUT LPS, YOU DON'T REALLY

2310
01:42:56,949 --> 01:42:59,451
SEE THAT MUCH BUT WITH LPS YOU

2311
01:42:59,451 --> 01:43:02,621
SEE ENHANCERS GET TURNED ON

2312
01:43:02,621 --> 01:43:05,891
AFTER LPS.

2313
01:43:05,891 --> 01:43:10,729
SO WE DID OUR LITTLE PDC OR

2314
01:43:10,729 --> 01:43:13,465
ARA-C SN 1 Seq, WITHOUT LPS WE

2315
01:43:13,465 --> 01:43:15,200
DON'T DETECT ANYTHING

2316
01:43:15,200 --> 01:43:15,501
WHATSOEVER.

2317
01:43:15,501 --> 01:43:20,439
BUT WITH LPS, WE CAN DETECT

2318
01:43:20,439 --> 01:43:22,674
THESE SINGLE-STRAND BREAKS

2319
01:43:22,674 --> 01:43:24,576
CONVERTED TO DOUBLE-STRAND

2320
01:43:24,576 --> 01:43:27,613
BREAKS, SO THESE ENHANCERS ARE

2321
01:43:27,613 --> 01:43:29,381
UNDERGOING ACTIVE DMEET LAYINGS

2322
01:43:29,381 --> 01:43:31,617
AND WE'RE CONVERTING THIS TO A

2323
01:43:31,617 --> 01:43:33,385
DOUBLE-STRAND BREAK SO WE CAN

2324
01:43:33,385 --> 01:43:34,286
DETECT IT.

2325
01:43:34,286 --> 01:43:37,189
AND WE CAN DO THIS ALSO JUST

2326
01:43:37,189 --> 01:43:38,524
WITH CELL BIOLOGY, WITH OUR

2327
01:43:38,524 --> 01:43:46,298
STAINING, WITH OUR BILL BONNER

2328
01:43:46,298 --> 01:43:48,167
GAMMA-H2AX, BONE MARROW DERIVED

2329
01:43:48,167 --> 01:43:49,068
MACROPHAGES AFTER ACA-C

2330
01:43:49,068 --> 01:43:50,602
TREATMENT, YOU DON'T SEE MUCH,

2331
01:43:50,602 --> 01:43:54,073
IF YOU HAVE LPS PLUS ARA-C YOU

2332
01:43:54,073 --> 01:43:57,142
GET NEW ENHANCERS UNDERGOING

2333
01:43:57,142 --> 01:43:57,843
ACTIVE DEMETHYLATION, TRIGGERED

2334
01:43:57,843 --> 01:43:59,812
A DNA DAMAGE RESPONSE, AND YOU

2335
01:43:59,812 --> 01:44:01,747
CAN SEE THESE SPOTS.

2336
01:44:01,747 --> 01:44:03,882
THESE LITTLE SPOTS ARE DEPENDENT

2337
01:44:03,882 --> 01:44:08,754
ON TDG BECAUSE IF YOU INACTIVATE

2338
01:44:08,754 --> 01:44:10,255
TDG YOU DON'T SEE THEM.

2339
01:44:10,255 --> 01:44:13,258
NOW WE CAN STUDY WHAT IS THE

2340
01:44:13,258 --> 01:44:17,096
ROLE OF ACTIVE DEMETHYLATION IN

2341
01:44:17,096 --> 01:44:17,963
A -- DURING TRANSCRIPTIONAL

2342
01:44:17,963 --> 01:44:18,464
RESPONSE.

2343
01:44:18,464 --> 01:44:19,932
WE STILL DON'T KNOW WHAT IT DOES

2344
01:44:19,932 --> 01:44:23,035
BUT WE KNOW THAT IT HAPPENS.

2345
01:44:23,035 --> 01:44:27,539
SO I JUST WANT TO END THERE.

2346
01:44:27,539 --> 01:44:31,743
AND THANK THE REALLY AMAZING

2347
01:44:31,743 --> 01:44:38,417
POSTDOCS THAT LED THIS PROJECT.

2348
01:44:38,417 --> 01:44:46,391
AND BEST HIRE I EVER MADE, ELSA

2349
01:44:46,391 --> 01:44:49,995
CALLEN IN MY LAB, GREAT

2350
01:44:49,995 --> 01:44:51,029
COLLABORATORS, AND OUR --

2351
01:44:51,029 --> 01:44:54,933
MICHAEL WARD WHO INTRODUCED ME

2352
01:44:54,933 --> 01:44:56,135
INTO THE SUBJECT OF NEURONAL

2353
01:44:56,135 --> 01:44:58,403
DAMAGE AND GOT ME INTERESTED IN

2354
01:44:58,403 --> 01:44:58,604
THIS.

2355
01:44:58,604 --> 01:44:59,771
I WANT TO THANK YOU.

2356
01:44:59,771 --> 01:45:02,407
[APPLAUSE]

2357
01:45:02,407 --> 01:45:05,844
 

2358
01:45:05,844 --> 01:45:07,613
>> APOLOGIES TO OUR ONLINE

2359
01:45:07,613 --> 01:45:09,181
VIEWERS, YOUR FIRST FIVE MINUTES

2360
01:45:09,181 --> 01:45:13,986
OF THE SLIDES WEREN'T SHOWING. 

2361
01:45:13,986 --> 01:45:15,721
AND THEIR URGENT MESSAGING TOOK

2362
01:45:15,721 --> 01:45:19,625
CARE OF THAT AFTER FIVE MINUTES.

2363
01:45:19,625 --> 01:45:21,660
I COULDN'T REPLY TO ALL AT ONCE.

2364
01:45:21,660 --> 01:45:23,395
>> AGAIN, THANK YOU SO
MUCH FOR

2365
01:45:23,395 --> 01:45:24,062
A WONDERFUL TALK.

2366
01:45:24,062 --> 01:45:27,266
I THINK ALL OF YOU ARE GETTING

2367
01:45:27,266 --> 01:45:31,003
THE SENSE OF THIS

2368
01:45:31,003 --> 01:45:32,938
INTERDISCIPLINARY NATURE OF WHAT

2369
01:45:32,938 --> 01:45:36,108
EVERYONE WHO HAS SPOKEN DOES AND

2370
01:45:36,108 --> 01:45:39,244
THE UNIQUE ENVIRONMENT OF THE

2371
01:45:39,244 --> 01:45:44,316
NIH THAT ALLOWS US TO REALLY

2372
01:45:44,316 --> 01:45:47,819
SEGUE BETWEEN FIELDS AND ORGAN

2373
01:45:47,819 --> 01:45:48,053
SYSTEMS.

2374
01:45:48,053 --> 01:45:49,988
NEXT SPEAKER IS NO EXCEPTION TO

2375
01:45:49,988 --> 01:45:52,658
THAT RULE. 

2376
01:45:52,658 --> 01:45:55,494
DR. JOHN O'SHEA, SCIENTIFIC

2377
01:45:55,494 --> 01:45:57,095
DIRECTOR AND SENIOR INVESTIGATOR

2378
01:45:57,095 --> 01:46:00,165
AT NIAMS.

2379
01:46:00,165 --> 01:46:07,873
HE WORKS ON CD4 POSITIVE T CELLS

2380
01:46:07,873 --> 01:46:08,974
AND CYTOKINES THEY PRODUCE,

2381
01:46:08,974 --> 01:46:10,943
EXPERT IN MECHANISMS OF NOT ONLY

2382
01:46:10,943 --> 01:46:12,844
THE PRODUCTION OF THOSE

2383
01:46:12,844 --> 01:46:16,515
CYTOKINES BUT ALSO IN THE SIGNAL

2384
01:46:16,515 --> 01:46:18,884
TRANSDUCTION PATHWAYS THAT THEY

2385
01:46:18,884 --> 01:46:22,754
INITIATE AT RECEPTORS, AND HE

2386
01:46:22,754 --> 01:46:26,191
STUDIES THOSE PATHWAYS IN

2387
01:46:26,191 --> 01:46:28,093
EFFORTS TO DESIGN THOUGHTFULLY

2388
01:46:28,093 --> 01:46:29,561
THERAPEUTIC INTERVENTIONS WHEN

2389
01:46:29,561 --> 01:46:31,830
THOSE PATHWAYS GO AWRY.

2390
01:46:31,830 --> 01:46:42,341
SO, JOHN, THE PODIUM IS YOURS.

2391
01:46:42,808 --> 01:46:43,008
[APPLAUSE]

2392
01:46:43,008 --> 01:46:45,510
>> THANKS, NINA.

2393
01:46:45,510 --> 01:46:50,349
 

2394
01:46:50,349 --> 01:46:51,016
LET'S SEE.

2395
01:46:51,016 --> 01:46:54,486
MOUSE HERE.

2396
01:46:54,486 --> 01:47:00,392
 

2397
01:47:00,392 --> 01:47:04,429
I JUST RUINED MY JOKE THERE.

2398
01:47:04,429 --> 01:47:05,430
OKAY.

2399
01:47:05,430 --> 01:47:08,300
THANKS VERY MUCH FOR THIS

2400
01:47:08,300 --> 01:47:08,600
OPPORTUNITY.

2401
01:47:08,600 --> 01:47:09,801
WHAT I WANTED TO DO IN THE NEXT

2402
01:47:09,801 --> 01:47:13,538
FEW MINUTES IS TALK ABOUT THE

2403
01:47:13,538 --> 01:47:19,111
DISCOVERY OF JAKs AND JAK

2404
01:47:19,111 --> 01:47:21,413
INHIBITORS AND IT'S BEEN ABOUT

2405
01:47:21,413 --> 01:47:23,615
30-YEAR SORT OF BIRTHDAY PARTY,

2406
01:47:23,615 --> 01:47:26,518
LAST YEAR, FOR THE JAK/STAT

2407
01:47:26,518 --> 01:47:28,186
PATHWAY, AND I WANTED TO REFLECT

2408
01:47:28,186 --> 01:47:32,357
ON WHAT WE NEED TO DO FOR THE 

2409
01:47:32,357 --> 01:47:35,527
NEXT 30 YEARS OR SO

2410
01:47:35,527 --> 01:47:37,629
I'LL TALK ABOUT THAT AND HOW

2411
01:47:37,629 --> 01:47:40,032
CYTOKINES MET THE GENOME, WHAT

2412
01:47:40,032 --> 01:47:41,833
THAT MEANS, COMMON AND RARE

2413
01:47:41,833 --> 01:47:46,038
DISEASES, SOME RARE DISEASES 

2414
01:47:46,038 --> 01:47:46,872
MAYBE NOT SO RARE.

2415
01:47:46,872 --> 01:47:49,341
I'M PLEASED I'M IN THE NATIONAL

2416
01:47:49,341 --> 01:47:51,977
ACADEMY BUT WHAT IS THE REAL

2417
01:47:51,977 --> 01:47:53,812
HONOR IN MANY RESPECTS IS JUST

2418
01:47:53,812 --> 01:47:55,247
BEING HERE AT THE NIH.

2419
01:47:55,247 --> 01:47:56,515
THERE'S SO MANY GREAT PEOPLE AND

2420
01:47:56,515 --> 01:47:59,918
I JUST THINK WHEN I CAME HERE,

2421
01:47:59,918 --> 01:48:02,154
JUST BLEW MY MIND, ALL THE

2422
01:48:02,154 --> 01:48:03,889
AMAZING SCIENTISTS AROUND ME.

2423
01:48:03,889 --> 01:48:07,859
I THINK I INCLUDED MOST PEOPLE'S

2424
01:48:07,859 --> 01:48:08,694
NAMES HERE.

2425
01:48:08,694 --> 01:48:11,229
BUT IF I LEFT OFF YOUR NAME,

2426
01:48:11,229 --> 01:48:15,300
JUST SEND ME A NOTE AND NEXT

2427
01:48:15,300 --> 01:48:17,102
TIME I HAVE THE OPPORTUNITY TO

2428
01:48:17,102 --> 01:48:22,040
PUT UP THIS SLIDE I PROMISE I'LL

2429
01:48:22,040 --> 01:48:23,108
ADD THAT.

2430
01:48:23,108 --> 01:48:26,578
BUT IT DOES MAKE ME THINK ABOUT

2431
01:48:26,578 --> 01:48:31,116
THIS AMAZING ECOSYSTEM THAT WE

2432
01:48:31,116 --> 01:48:31,616
HAVE.

2433
01:48:31,616 --> 01:48:33,085
MICHAEL, I THINK FOR A LOT OF

2434
01:48:33,085 --> 01:48:36,555
TIME I WAS -- YOU KNOW, STARTING

2435
01:48:36,555 --> 01:48:38,190
OFF AS, YOU KNOW, I THINK YOU'RE

2436
01:48:38,190 --> 01:48:40,258
RESPONSIBLE FOR A LOT OF THIS

2437
01:48:40,258 --> 01:48:40,592
ECOSYSTEM.

2438
01:48:40,592 --> 01:48:42,994
NINA, I DON'T MEAN TO MAKE YOU

2439
01:48:42,994 --> 01:48:44,596
FEEL NERVOUS BY ANY MEANS BUT

2440
01:48:44,596 --> 01:48:46,898
IT'S REALLY -- YOU SAID THE SAME

2441
01:48:46,898 --> 01:48:49,301
THING, MICHAEL GETS CREDIT FOR

2442
01:48:49,301 --> 01:48:50,836
THIS?

2443
01:48:50,836 --> 01:48:51,436
YEAH.

2444
01:48:51,436 --> 01:48:52,804
JUST AN AMAZING PLACE.

2445
01:48:52,804 --> 01:48:55,140
AND REALLY IN MANY RESPECTS

2446
01:48:55,140 --> 01:48:57,008
THAT'S REALLY THE HONOR.

2447
01:48:57,008 --> 01:48:58,944
SO, HOW DID I GET STARTED IN

2448
01:48:58,944 --> 01:48:59,578
THIS BUSINESS?

2449
01:48:59,578 --> 01:49:02,681
I WAS A MEDICAL STUDENT, DOING

2450
01:49:02,681 --> 01:49:04,116
ROTATION IN INFECTIOUS DISEASE.

2451
01:49:04,116 --> 01:49:05,650
I SAW A PATIENT AT THE TIME THAT

2452
01:49:05,650 --> 01:49:07,486
HAD THE DISEASE THAT WE DON'T

2453
01:49:07,486 --> 01:49:10,422
CALL SO MUCH ANYMORE BUT ACUTE

2454
01:49:10,422 --> 01:49:11,022
RESPIRATORY DISTRESS SYNDROME

2455
01:49:11,022 --> 01:49:12,724
WAS NEW AT THE TIME.

2456
01:49:12,724 --> 01:49:15,093
THE ATTENDING TOLD ME THAT YOU

2457
01:49:15,093 --> 01:49:18,897
COULD TAKE THESE PATIENTS IN

2458
01:49:18,897 --> 01:49:23,068
SEPTIC SHOCK, GIVE THEM

2459
01:49:23,068 --> 01:49:24,102
ANTIBIOTICS, STERILE BLOOD

2460
01:49:24,102 --> 01:49:25,537
CULTURES BUT SOMETHING CAUSED

2461
01:49:25,537 --> 01:49:27,639
THIS INFLAMMATION, I DIDN'T KNOW

2462
01:49:27,639 --> 01:49:30,575
MUCH ABOUT THAT BUT HE THEN

2463
01:49:30,575 --> 01:49:33,111
STILL -- I CAN SEE THE POINTER,

2464
01:49:33,111 --> 01:49:38,483
MAYBE I'LL DO IT THIS WAY.

2465
01:49:38,483 --> 01:49:38,683
OKAY.

2466
01:49:38,683 --> 01:49:39,317
I'LL DO THAT.

2467
01:49:39,317 --> 01:49:41,086
WHAT HE HAD BEEN WORKING ON FOR

2468
01:49:41,086 --> 01:49:47,192
A VERY LONG TIME WERE THINGS

2469
01:49:47,192 --> 01:49:48,527
CALLED LEUKOCYTE ENDOGENOUS

2470
01:49:48,527 --> 01:49:48,794
PYROGENS.

2471
01:49:48,794 --> 01:49:50,629
I SAID CAN I JUST TAKE SOME TIME

2472
01:49:50,629 --> 01:49:52,230
AND WORK IN YOUR LAB?

2473
01:49:52,230 --> 01:49:56,902
I WORKED IN HIS LAB WHERE HE WAS

2474
01:49:56,902 --> 01:49:58,036
TRYING TO PURIFY WHAT ULTIMATELY

2475
01:49:58,036 --> 01:50:01,440
TURNED OUT TO BE INTERLEUKIN 1

2476
01:50:01,440 --> 01:50:04,276
AND TNF AND IL-6 AND OTHERS, HE

2477
01:50:04,276 --> 01:50:08,213
DIDN'T SUCCEED IN THAT BUT AS

2478
01:50:08,213 --> 01:50:10,415
YOU KNOW CHARLES WHO WAS HERE

2479
01:50:10,415 --> 01:50:16,488
AND MOVED ON AND EGAL GEARY DID

2480
01:50:16,488 --> 01:50:18,457
DISCOVER IL-1.

2481
01:50:18,457 --> 01:50:19,491
THE OTHER THING THAT HAPPENED

2482
01:50:19,491 --> 01:50:21,126
LATER WHEN I WAS A RESIDENT I

2483
01:50:21,126 --> 01:50:24,062
SAW A PATIENT WITH VASCULITIS.

2484
01:50:24,062 --> 01:50:25,630
THIS IS PETER GRAYSON'S IMAGE

2485
01:50:25,630 --> 01:50:28,400
HERE, BUT THAT'S -- WE OBVIOUSLY

2486
01:50:28,400 --> 01:50:29,634
DIDN'T HAVE THIS TECHNOLOGY BACK

2487
01:50:29,634 --> 01:50:32,137
WHEN I WAS SEEING THIS PATIENT.

2488
01:50:32,137 --> 01:50:33,772
BUT I THEN IMMEDIATELY WENT TO

2489
01:50:33,772 --> 01:50:36,041
THE LIBRARY AND THEN HAD TO READ

2490
01:50:36,041 --> 01:50:38,243
UP ON VASCULITIS AND I CAME

2491
01:50:38,243 --> 01:50:42,614
ACROSS THIS GUY HERE WHO YOU MAY

2492
01:50:42,614 --> 01:50:44,249
KNOW, TONY FAUCI, ALL OF US AT

2493
01:50:44,249 --> 01:50:47,519
THE TIME WOULD HAVE OUR WHITE

2494
01:50:47,519 --> 01:50:50,188
LAB COATS, SPECTRUM OF

2495
01:50:50,188 --> 01:50:50,589
VASCULITIS.

2496
01:50:50,589 --> 01:50:52,524
I CALLED TONY AND COLLEAGUES,

2497
01:50:52,524 --> 01:50:54,493
WHAT DO I DO WITH THIS PATIENT?

2498
01:50:54,493 --> 01:50:57,629
WHAT WAS REALLY NEW AT THE TIME

2499
01:50:57,629 --> 01:51:02,501
WAS TREATMENT OF AUTOIMMUNE 

2500
01:51:02,501 --> 01:51:03,068
DISEASES WITH CANCER DRUGS,

2501
01:51:03,068 --> 01:51:05,036
CYCLOPHOSPHAMIDE, ET CETERA.

2502
01:51:05,036 --> 01:51:07,005
I GOT SO INTERESTED I WROTE TO

2503
01:51:07,005 --> 01:51:08,406
TONY, ANY CHANCE YOU COULD PICK

2504
01:51:08,406 --> 01:51:12,777
ME UP AND HAVE ME JOIN NIAID?

2505
01:51:12,777 --> 01:51:19,551
WHICH TURNED OUT TO BE THE CASE.

2506
01:51:19,551 --> 01:51:22,187
I CAME TO THE NIH, BUT TONY AT

2507
01:51:22,187 --> 01:51:24,656
THAT POINT MOVED ON TO ANOTHER

2508
01:51:24,656 --> 01:51:27,526
DISEASE WORKING ON HIV NOT SO

2509
01:51:27,526 --> 01:51:28,159
MUCH VASCULITIS.

2510
01:51:28,159 --> 01:51:32,197
I WAS MORE INTERESTED IN

2511
01:51:32,197 --> 01:51:33,498
CYTOKINE -- IN IMMUNE RECEPTORS,

2512
01:51:33,498 --> 01:51:36,568
AND STARTED OFF WITH COMPLEMENT

2513
01:51:36,568 --> 01:51:38,537
RECEPTORS BUT THEN WORKED WITH

2514
01:51:38,537 --> 01:51:42,274
BILL PAUL AND RICK CLAUSENER,

2515
01:51:42,274 --> 01:51:44,209
LARRY SAMUELSON TO WORK ON OTHER

2516
01:51:44,209 --> 01:51:46,077
RECEPTORS AS WELL, AND AGAIN

2517
01:51:46,077 --> 01:51:49,114
JUST AN AMAZING TIME AT THE NIH

2518
01:51:49,114 --> 01:51:52,350
TO SEE DISCOVERY AFTER STOVER.

2519
01:51:52,350 --> 01:51:54,553
AND SO WHEN I HAD STARTED MY OWN

2520
01:51:54,553 --> 01:51:56,254
LAB I STARTED THINKING ABOUT THE

2521
01:51:56,254 --> 01:52:00,492
ONE THING RICK TAUGHT ME ABOUT

2522
01:52:00,492 --> 01:52:01,426
WAS THAT TYROSINE KINASES SEEM

2523
01:52:01,426 --> 01:52:05,096
TO BE REALLY IMPORTANT FOR THE

2524
01:52:05,096 --> 01:52:06,498
PROXIMAL EVENTS IN SIGNAL

2525
01:52:06,498 --> 01:52:09,734
TRANSDUCTION BY LOTS OF THINGS,

2526
01:52:09,734 --> 01:52:11,169
FROM INSULIN TO GROWTH FACTORS,

2527
01:52:11,169 --> 01:52:11,970
ET CETERA.

2528
01:52:11,970 --> 01:52:15,507
WHAT WAS REVEALED WAS THAT

2529
01:52:15,507 --> 01:52:16,474
CYTOKINES ALSO INDUCED TYROSINE

2530
01:52:16,474 --> 01:52:17,609
PHOSPHORYLATION, BUT THE RACE

2531
01:52:17,609 --> 01:52:20,679
WAS ON, WHAT IS THE KINASE

2532
01:52:20,679 --> 01:52:22,914
THAT'S RESPONSIBLE FOR THIS.

2533
01:52:22,914 --> 01:52:25,483
AND SO WE DID WHAT A LOT OF

2534
01:52:25,483 --> 01:52:26,585
PEOPLE DID.

2535
01:52:26,585 --> 01:52:28,620
WE GENERATED LIBRARIES LOOKING

2536
01:52:28,620 --> 01:52:31,456
FOR KINASES IN MY CASE IN IMMUNE

2537
01:52:31,456 --> 01:52:31,823
CELLS.

2538
01:52:31,823 --> 01:52:33,858
AND REALLY THE KEY THING THAT

2539
01:52:33,858 --> 01:52:37,896
HAPPENED NEXT WAS THIS WORK BY

2540
01:52:37,896 --> 01:52:42,367
GEORGE STARK AND ALSO JIM

2541
01:52:42,367 --> 01:52:42,667
DARNELL.

2542
01:52:42,667 --> 01:52:44,869
JIM WAS A YELLOW BERET HERE AT

2543
01:52:44,869 --> 01:52:47,572
THE NIH AND HAD LOTS OF STORIES

2544
01:52:47,572 --> 01:52:50,175
ABOUT HIS TIME AT THE NIH.

2545
01:52:50,175 --> 01:52:54,679
BUT WHAT GEORGE DID WAS TO TAKE

2546
01:52:54,679 --> 01:52:56,948
CELLS AND MUTAGENIZE THEM AND

2547
01:52:56,948 --> 01:52:59,784
IDENTIFY CELLS THAT LACKED THEIR

2548
01:52:59,784 --> 01:53:02,354
SENSITIVITY TO INTERFERENCE AND

2549
01:53:02,354 --> 01:53:04,823
SANDRA PELEGRINEY REALIZED SHE

2550
01:53:04,823 --> 01:53:06,758
COULD COMPLEMENT CELLS WITH THIS

2551
01:53:06,758 --> 01:53:12,731
KINASE HERE WITH THIS AMBIGUOUS

2552
01:53:12,731 --> 01:53:15,667
NAME, TURNS OUT TO BE A JAK, A

2553
01:53:15,667 --> 01:53:16,534
KEY DISCOVERY.

2554
01:53:16,534 --> 01:53:21,473
JIM IS TAKING A DIVERSITY

2555
01:53:21,473 --> 01:53:24,843
APPROACH USING THESE ELEMENTS AS

2556
01:53:24,843 --> 01:53:28,813
BAIT TO DISCOVER STAT FAMILY OF

2557
01:53:28,813 --> 01:53:29,381
TRANSCRIPTION FACTORS.

2558
01:53:29,381 --> 01:53:33,852
SO, THIS IS AN AMAZING TOOL, ALL

2559
01:53:33,852 --> 01:53:35,787
THESE MUTAGENIZED CELLS, VERY

2560
01:53:35,787 --> 01:53:39,190
QUICKLY WE BEGAN TO SORT OF PUT

2561
01:53:39,190 --> 01:53:42,127
TOGETHER THE PATHWAYS OF

2562
01:53:42,127 --> 01:53:43,361
CYTOKINES, CYTOKINE RECEPTORS,

2563
01:53:43,361 --> 01:53:45,296
DIFFERENT JAKs, ET CETERA.

2564
01:53:45,296 --> 01:53:48,700
NOW TURNED OUT THAT THERE ARE 57

2565
01:53:48,700 --> 01:53:52,570
CYTOKINES THAT USE JAK/STAT 

2566
01:53:52,570 --> 01:53:54,939
PATHWAY, PATHWAY IS CONSERVED

2567
01:53:54,939 --> 01:53:56,374
FROM INSECTS TO MAMMALS.

2568
01:53:56,374 --> 01:53:57,776
BUT THE KEY QUESTION REALLY WAS

2569
01:53:57,776 --> 01:54:02,480
WHAT DO WE KNOW ABOUT THE IN

2570
01:54:02,480 --> 01:54:06,551
VIVO CRITICALITY OF THIS

2571
01:54:06,551 --> 01:54:08,620
JAK/STAT PATHWAY.

2572
01:54:08,620 --> 01:54:11,690
WE MADE KNOCKOUT MICE, BERG AND

2573
01:54:11,690 --> 01:54:14,392
OTHERS, BUT THE FIRST JAK/STAT

2574
01:54:14,392 --> 01:54:16,695
KNOCKOUT CAME IN SLIGHTLY

2575
01:54:16,695 --> 01:54:18,463
DIFFERENT WAY, AND IT STARTS

2576
01:54:18,463 --> 01:54:25,136
WITH THIS COLLABORATION WITH

2577
01:54:25,136 --> 01:54:26,504
WARREN LEONARD.

2578
01:54:26,504 --> 01:54:28,540
JAK 3 IS HIGHLY EXPRESSED IN

2579
01:54:28,540 --> 01:54:29,441
IMMUNE CELLS, SUSPICIOUS IT

2580
01:54:29,441 --> 01:54:34,879
MIGHT BE INVOLVED IN SIGNALING

2581
01:54:34,879 --> 01:54:36,715
VIA THESE CYTOKINES HERE THAT

2582
01:54:36,715 --> 01:54:38,516
USE COMMON GAMMA CHAIN.

2583
01:54:38,516 --> 01:54:41,619
WHAT WARREN HAD SHOWN WAS THAT

2584
01:54:41,619 --> 01:54:44,322
MUTATIONS OF THE COMMON GAMMA

2585
01:54:44,322 --> 01:54:46,191
CHAIN SHOWN HERE UNDERLIE THE

2586
01:54:46,191 --> 01:54:47,058
DISEASE, BUBBLE BOY SYNDROME,

2587
01:54:47,058 --> 01:54:50,328
AND IF YOU HAD THESE MUTATIONS

2588
01:54:50,328 --> 01:54:53,264
YOU COULDN'T SIGNAL VIA

2589
01:54:53,264 --> 01:54:53,698
CYTOKINES.

2590
01:54:53,698 --> 01:54:56,701
SO WITH WARREN, WE SPECULATED

2591
01:54:56,701 --> 01:54:58,903
AND THEN PROVED THAT JAK 3

2592
01:54:58,903 --> 01:54:59,704
ASSOCIATES WITH COMMON GAMMA

2593
01:54:59,704 --> 01:55:01,272
CHAIN, IN THE SAME PAPER SAID,

2594
01:55:01,272 --> 01:55:03,441
WELL, WAIT A MINUTE, IF YOU HAVE

2595
01:55:03,441 --> 01:55:05,877
MUTATIONS OF GAMMA C MAYBE IF

2596
01:55:05,877 --> 01:55:07,645
YOU HAVE MUTATIONS OF JAK 3

2597
01:55:07,645 --> 01:55:13,918
YOU'LL GET A SIMILAR PHENOTYPE.

2598
01:55:13,918 --> 01:55:16,121
IN FACT GIGI NOTARANGELO ACTED

2599
01:55:16,121 --> 01:55:19,958
ON THAT AND LOOKED AT HIS COHORT

2600
01:55:19,958 --> 01:55:22,026
OF PATIENTS.

2601
01:55:22,026 --> 01:55:24,095
FABIO CANDOTTI, WHO CARED FOR

2602
01:55:24,095 --> 01:55:26,064
SOME OF THESE PATIENTS IN GIGI'S

2603
01:55:26,064 --> 01:55:28,900
COHORT WAS WORKING IN MY LAB AT

2604
01:55:28,900 --> 01:55:30,435
THAT TIME.

2605
01:55:30,435 --> 01:55:33,271
ANOTHER AMAZING PART ABOUT BEING

2606
01:55:33,271 --> 01:55:34,405
THEY NIH.

2607
01:55:34,405 --> 01:55:37,442
AND SO THEN WARREN COLLABORATED

2608
01:55:37,442 --> 01:55:38,276
WITH REBECCA BUCKLEY'S GROUP,

2609
01:55:38,276 --> 01:55:40,745
AND WHAT WE SHOWED WAS THAT

2610
01:55:40,745 --> 01:55:45,483
MUTATIONS OF JAK 3 UNDERLIE AND 

2611
01:55:45,483 --> 01:55:50,021
FORM SCID THAT'S SIMILAR TO

2612
01:55:50,021 --> 01:55:50,488
X-LINK SEVERE COMBINED

2613
01:55:50,488 --> 01:55:51,790
IMMUNODEFICIENCY.

2614
01:55:51,790 --> 01:55:54,592
THIS IS THE FIRST KNOCK OUTIN

2615
01:55:54,592 --> 01:55:57,962
THE JAK/STAT PATHWAY, VERY

2616
01:55:57,962 --> 01:56:01,666
QUICKLY OTHER KNOCKOUT MICE

2617
01:56:01,666 --> 01:56:02,367
CONFIRMED THESE FINDINGS.

2618
01:56:02,367 --> 01:56:05,537
THE OTHER PART ABOUT THIS IS

2619
01:56:05,537 --> 01:56:09,707
THAT THE AMAZING PART ABOUT THIS

2620
01:56:09,707 --> 01:56:10,508
STORY WITH THE CYTOKINE

2621
01:56:10,508 --> 01:56:12,410
REVOLUTION WITH ALL THE CLONING

2622
01:56:12,410 --> 01:56:14,479
OF ALL THE MANY CYTOKINES THAT

2623
01:56:14,479 --> 01:56:16,181
WE HAVE IN THE GENOME PROBABLY

2624
01:56:16,181 --> 01:56:18,883
MORE THAN 200 OR SO THAT WAS

2625
01:56:18,883 --> 01:56:21,953
RAPIDLY CAPITALIZED UPON BY

2626
01:56:21,953 --> 01:56:24,656
MAKING A VARIETY OF MONOCLONAL

2627
01:56:24,656 --> 01:56:27,625
ANTIBODIES, THE NEXT LEVEL

2628
01:56:27,625 --> 01:56:31,930
BEYOND DRUGS LIKE METHOTREXATE

2629
01:56:31,930 --> 01:56:33,097
AND CYCLOPHOSPHAMIDE, ET CETERA.

2630
01:56:33,097 --> 01:56:35,733
WE HAD THE OPPORTUNITY TO REALLY

2631
01:56:35,733 --> 01:56:39,404
BEGIN TO SHUT OFF THE

2632
01:56:39,404 --> 01:56:40,405
INFLAMMATORY RESPONSES BY

2633
01:56:40,405 --> 01:56:41,706
TARGETING TNF, ET CETERA, ET

2634
01:56:41,706 --> 01:56:43,441
CETERA, ALL THE OTHER DIFFERENT

2635
01:56:43,441 --> 01:56:46,878
CYTOKINES SHOWN HERE BUT DESPITE

2636
01:56:46,878 --> 01:56:49,380
THAT, DESPITE THE IMPRESSIVE

2637
01:56:49,380 --> 01:56:50,815
RESULTS WITH BIOLOGICS, THERE

2638
01:56:50,815 --> 01:56:53,017
ARE PATIENTS WHO HAD INCOMPLETE

2639
01:56:53,017 --> 01:56:54,352
OR DIDN'T REALLY RESPOND TO

2640
01:56:54,352 --> 01:56:55,887
THESE DRUGS.

2641
01:56:55,887 --> 01:56:57,021
WHAT OTHER OPTIONS MIGHT WE

2642
01:56:57,021 --> 01:56:57,222
HAVE?

2643
01:56:57,222 --> 01:57:00,859
THE IDEA AT THE TIME WAS CAN WE

2644
01:57:00,859 --> 01:57:03,394
TARGET SIGNALING MOLECULES

2645
01:57:03,394 --> 01:57:11,469
MAINLY CAN WE TARGET JAKs?

2646
01:57:11,469 --> 01:57:13,838
IS WE MADE THIS BOLD CLAIM WITH

2647
01:57:13,838 --> 01:57:15,607
WARREN IN 1995.

2648
01:57:15,607 --> 01:57:18,009
WE SAID THIS WORK SUGGESTS WE

2649
01:57:18,009 --> 01:57:20,645
COULD TARGET JAKs, A NEW CLASS

2650
01:57:20,645 --> 01:57:21,446
OF IMMUNOSUPPRESSANT MOLECULES,

2651
01:57:21,446 --> 01:57:23,248
BUT AS YOU MIGHT IMAGINE, IT'S

2652
01:57:23,248 --> 01:57:25,516
ALWAYS EASY TO MAKE A PREDICTION

2653
01:57:25,516 --> 01:57:27,852
WHEN YOU KNOW THE ANSWER.

2654
01:57:27,852 --> 01:57:29,888
SO THIS IS IN 1995, BUT I WAS

2655
01:57:29,888 --> 01:57:33,091
FORTUNATE ENOUGH TO BE AT THIS

2656
01:57:33,091 --> 01:57:36,160
MEETING HERE IN LOVELY SAXTON

2657
01:57:36,160 --> 01:57:37,562
RIVERS, VERMONT, IN 1993.

2658
01:57:37,562 --> 01:57:42,700
I RAN INTO THIS GUY, PAUL

2659
01:57:42,700 --> 01:57:44,669
CHINGELLI, WE WORKED ON

2660
01:57:44,669 --> 01:57:46,905
COMPLEMENT RECEPTORS, SORT OF

2661
01:57:46,905 --> 01:57:48,273
COMPETITORS, HE WAS AT HARVARD,

2662
01:57:48,273 --> 01:57:50,808
I WAS WORKING WITH MIKE FRANK.

2663
01:57:50,808 --> 01:57:55,613
BUT IT WAS A FRIENDLY

2664
01:57:55,613 --> 01:57:55,914
COMPETITION.

2665
01:57:55,914 --> 01:57:56,848
BUT I SAID AT THE TIME WAIT A

2666
01:57:56,848 --> 01:57:59,217
MINUTE, PAUL, WHAT ARE YOU DOING

2667
01:57:59,217 --> 01:58:00,785
HERE AT THIS LYMPHOCYTE AND

2668
01:58:00,785 --> 01:58:01,986
ANTIBODIES MEETING?

2669
01:58:01,986 --> 01:58:06,824
I'M WORKING AT AT PFIZER, NOW,

2670
01:58:06,824 --> 01:58:11,396
AND PFIZER IS THINKING WE SHOULD

2671
01:58:11,396 --> 01:58:13,164
TARGET TYROSINE KINASE AS NEW

2672
01:58:13,164 --> 01:58:14,432
THERAPY FOR AUTOIMMUNE DISEASE.

2673
01:58:14,432 --> 01:58:18,703
HE'S RIGHT NEXT TO ART WEISS,

2674
01:58:18,703 --> 01:58:21,372
WHO JUST CLONED JAK 7, I THINK

2675
01:58:21,372 --> 01:58:22,907
HE WAS TRYING TO GET AWAY FROM

2676
01:58:22,907 --> 01:58:26,911
ME AND TALK TO ART.

2677
01:58:26,911 --> 01:58:27,278
OOPS.

2678
01:58:27,278 --> 01:58:29,414
I'M RUINING MY JOKES HERE.

2679
01:58:29,414 --> 01:58:29,681
[LAUGHTER]

2680
01:58:29,681 --> 01:58:31,182
IT'S OKAY.

2681
01:58:31,182 --> 01:58:33,718
YOU LAUGHED, THAT'S GOOD.

2682
01:58:33,718 --> 01:58:36,554
SO, YOU COULD ALSO SEE HERE ALL

2683
01:58:36,554 --> 01:58:47,031
SORTS OF OTHER NIH-ERS HERE,

2684
01:58:48,666 --> 01:58:51,202
MIKE LEOARDO AND ON AND ON.

2685
01:58:51,202 --> 01:58:55,606
WE HAD GONE INTO THE WOODS HERE,

2686
01:58:55,606 --> 01:59:03,982
WE HAD THIS COLLABORATION 

2687
01:59:03,982 --> 01:59:05,783
CRADA, WHAT WOULD WE DID?

2688
01:59:05,783 --> 01:59:06,684
THAT'S A FORBES MAGAZINE WHETHER

2689
01:59:06,684 --> 01:59:11,456
THEY TALK ABOUT THE IDEA OF THIS

2690
01:59:11,456 --> 01:59:13,591
COLLABORATION BETWEEN PUBLIC AND

2691
01:59:13,591 --> 01:59:14,459
PRIVATE APPROACH WHICH, YOU

2692
01:59:14,459 --> 01:59:17,695
KNOW, LOTS TO SAY THERE BUT

2693
01:59:17,695 --> 01:59:19,430
MAYBE IN A DIFFERENT LECTURE.

2694
01:59:19,430 --> 01:59:22,867
IN ANY CASE, NOW THIS IS WHERE

2695
01:59:22,867 --> 01:59:24,235
WE ARE NOW.

2696
01:59:24,235 --> 01:59:27,605
THERE ARE 11 APPROVED

2697
01:59:27,605 --> 01:59:30,742
INDICATIONS FOR JAK INHIBITORS, 

2698
01:59:30,742 --> 01:59:32,176
12 JAK INHIBITORS, MANY MORE ON

2699
01:59:32,176 --> 01:59:34,112
THE WAY, DRUGS APPROVED FOR

2700
01:59:34,112 --> 01:59:39,717
VARIOUS FORMS OF ARTHRITIS SHOWN

2701
01:59:39,717 --> 01:59:41,919
HERE. 

2702
01:59:41,919 --> 01:59:43,321
ULCERATIVE COLITIS,

2703
01:59:43,321 --> 01:59:47,992
CORONAVIRUS'S DISEASE,

2704
01:59:47,992 --> 01:59:49,227
DERMATOLOGICAL DISEASES AND JAK

2705
01:59:49,227 --> 01:59:50,661
INHIBITORS USEFUL IN CYTOKINE

2706
01:59:50,661 --> 01:59:53,064
STORM ASSOCIATED WITH COVID-19.

2707
01:59:53,064 --> 01:59:54,732
WELL, IT'S INTERESTING IN THAT

2708
01:59:54,732 --> 01:59:56,901
REGARD THAT WITH THE CRADA WHEN

2709
01:59:56,901 --> 01:59:59,103
WE WERE WORKING WITH PFIZER, YOU

2710
01:59:59,103 --> 02:00:01,072
COULD IMAGINE THAT THERE'S SOME

2711
02:00:01,072 --> 02:00:03,241
RESTRICTIONS ON WE DO THESE

2712
02:00:03,241 --> 02:00:10,248
EXPERIMENTS AND I REMEMBER WE 

2713
02:00:10,248 --> 02:00:18,056
SAID CAN WE USE TOFACINITIB?

2714
02:00:18,056 --> 02:00:27,432
I LIKED THE  EXPERIMENT.

2715
02:00:27,432 --> 02:00:29,367
COLLABORATED WITH NCATS, HOME

2716
02:00:29,367 --> 02:00:36,507
BREW, WE SHOWED IN SEPTIC SHOCK

2717
02:00:36,507 --> 02:00:39,243
MODEL JAK INHIBITORS BLOCKED

2718
02:00:39,243 --> 02:00:41,479
THAT SEPTIC SHOCK PHENOTYPE,

2719
02:00:41,479 --> 02:00:47,652
USEFUL KNOWLEDGE DURING THE

2720
02:00:47,652 --> 02:00:49,754
PANDEMIC.

2721
02:00:49,754 --> 02:00:54,592
ELEVEN  JAK INHIBITORS, NOT JUST

2722
02:00:54,592 --> 02:00:56,794
FOR PEOPLE BUT ALSO FOR DOGS.

2723
02:00:56,794 --> 02:00:58,329
THIS IS AN NIH DOG.

2724
02:00:58,329 --> 02:01:00,598
THIS IS THE JOKE I RUINED.

2725
02:01:00,598 --> 02:01:01,799
I'VE SHOWN THIS MANY TIMES.

2726
02:01:01,799 --> 02:01:04,969
WHEN I GO AROUND THE WORLD AND

2727
02:01:04,969 --> 02:01:06,137
SHOW PICTURES OF JAK INHIBITORS,

2728
02:01:06,137 --> 02:01:07,772
ET CETERA, ET CETERA, PEOPLE

2729
02:01:07,772 --> 02:01:09,173
HAVE A TENDENCY TO SHOW ME

2730
02:01:09,173 --> 02:01:13,611
PICTURES OF THEIR DOG SO IF

2731
02:01:13,611 --> 02:01:17,849
ANYBODY'S DOGS ARE ON A JAK

2732
02:01:17,849 --> 02:01:19,717
INHIBITORS, SEND ME A PICTURE.

2733
02:01:19,717 --> 02:01:22,120
I'LL PRESERVE, THESE ARE HIPAA

2734
02:01:22,120 --> 02:01:26,124
COMPLIANT DOGS BUT HAPPY TO SHOW

2735
02:01:26,124 --> 02:01:27,091
THESE PICTURES.

2736
02:01:27,091 --> 02:01:30,261
THIS IS WHERE WE WERE WITH

2737
02:01:30,261 --> 02:01:31,929
BIOLOGICS, TURN THE LIGHT ON,

2738
02:01:31,929 --> 02:01:34,298
TURN THE LIGHT OFF, YOU HAVE A

2739
02:01:34,298 --> 02:01:35,566
DISEASE ASSOCIATED WITH

2740
02:01:35,566 --> 02:01:36,968
CYTOKINES, BUT THEN JAK

2741
02:01:36,968 --> 02:01:39,637
INHIBITORS GAVE THE OPPORTUNITY

2742
02:01:39,637 --> 02:01:42,807
TO HAVE A RHEOSTAT, GETTING FROM

2743
02:01:42,807 --> 02:01:43,574
HOME DEPOT.

2744
02:01:43,574 --> 02:01:45,476
IT GIVES TWO OPTIONS TO BE

2745
02:01:45,476 --> 02:01:49,780
THINKING ABOUT HOW TO TREAT

2746
02:01:49,780 --> 02:01:50,148
THESE DISEASES.

2747
02:01:50,148 --> 02:01:54,485
BUT THE OTHER THING WE'RE

2748
02:01:54,485 --> 02:01:55,853
THINKING ABOUT WAS WHAT'S AHEAD

2749
02:01:55,853 --> 02:01:57,922
FOR THE NEXT 30 YEARS?

2750
02:01:57,922 --> 02:02:01,058
OBVIOUSLY WE HAVE A LOT TO DO

2751
02:02:01,058 --> 02:02:01,759
WITH UNDERSTANDING DISEASE

2752
02:02:01,759 --> 02:02:07,932
MECHANISM, MUCH BETTER, HAVE A

2753
02:02:07,932 --> 02:02:09,834
BETTER UNDERSTANDING OF

2754
02:02:09,834 --> 02:02:10,735
GENOMICS, PERSONALIZED MEDICINE,

2755
02:02:10,735 --> 02:02:12,470
CAPITALIZE ON GENE EXPRESSION.

2756
02:02:12,470 --> 02:02:15,406
THE OTHER THING THANKS TO CHRIS

2757
02:02:15,406 --> 02:02:17,475
GARCIA'S WORK, NOW JUST

2758
02:02:17,475 --> 02:02:20,178
RECENTLY, IN THE LAST YEAR OR

2759
02:02:20,178 --> 02:02:22,146
SO, WE FINALLY HAVE THE

2760
02:02:22,146 --> 02:02:25,816
STRUCTURE OF JAK SO THE FIRST

2761
02:02:25,816 --> 02:02:28,386
GENERATION OF JAK INHIBITORS

2762
02:02:28,386 --> 02:02:31,055
TARGETED THE CATALYTIC DOMAIN,

2763
02:02:31,055 --> 02:02:33,424
SO COMPETITIVE ANTAGONIST OF

2764
02:02:33,424 --> 02:02:33,991
ATP.

2765
02:02:33,991 --> 02:02:35,092
I REMEMBER WHEN PAUL WAS TELLING

2766
02:02:35,092 --> 02:02:38,496
ME ABOUT THIS, I WAS THINKING

2767
02:02:38,496 --> 02:02:39,096
ATP?

2768
02:02:39,096 --> 02:02:40,264
YOU'RE GOING TO TARGET ATP?

2769
02:02:40,264 --> 02:02:41,766
THAT'S NOT POSSIBLE.

2770
02:02:41,766 --> 02:02:45,002
IT IS POSSIBLE.

2771
02:02:45,002 --> 02:02:46,370
BUT IT'S ACTUALLY PROBABLY IF

2772
02:02:46,370 --> 02:02:52,410
YOU WANT TO GET MORE 

2773
02:02:52,410 --> 02:02:56,914
SPECIFICITY, TARGETING GETTING

2774
02:02:56,914 --> 02:02:57,648
ALLOSTERIC INHIBITORS.

2775
02:02:57,648 --> 02:03:01,018
I WANTED TO GO BACK TO THINKING

2776
02:03:01,018 --> 02:03:03,187
ABOUT DISEASE MECHANISM AND THEN

2777
02:03:03,187 --> 02:03:03,921
PERSONALIZED MEDICINE, ET

2778
02:03:03,921 --> 02:03:05,022
CETERA, AND GENE EXPRESSION.

2779
02:03:05,022 --> 02:03:08,659
I'LL GIVE A FEW EXAMPLES OF

2780
02:03:08,659 --> 02:03:08,859
THAT.

2781
02:03:08,859 --> 02:03:12,563
SO, THIS IS THE CARTOON I SHOWED

2782
02:03:12,563 --> 02:03:13,864
JUST BEFORE, THINKING ABOUT WAY

2783
02:03:13,864 --> 02:03:17,768
BACK IN THE 20th CENTURY, THIS

2784
02:03:17,768 --> 02:03:20,137
IS HOW WE THOUGHT ABOUT

2785
02:03:20,137 --> 02:03:22,406
SIGNALING, PROMOTER CENTRIC,

2786
02:03:22,406 --> 02:03:23,507
THINKING ABOUT TRANSCRIPTION

2787
02:03:23,507 --> 02:03:24,709
FACTORS, BINDING THESE PORTIONS

2788
02:03:24,709 --> 02:03:26,577
OF THE GENOME BUT ONE THING THAT

2789
02:03:26,577 --> 02:03:30,114
WE FOUND IS THAT THE GENOME IS

2790
02:03:30,114 --> 02:03:34,385
ONLY 2% GENES AND 98% OF OTHER

2791
02:03:34,385 --> 02:03:36,420
STUFF, AND AT THE TIME WE DIDN'T

2792
02:03:36,420 --> 02:03:45,096
REALLY KNOW WHAT THIS STUFF WAS.

2793
02:03:45,096 --> 02:03:45,930
THAT GOES BEYOND PROMOTERS.

2794
02:03:45,930 --> 02:03:49,033
MUCH OF THE GENOME IS ACTIVE IN

2795
02:03:49,033 --> 02:03:51,168
CELL-SPECIFIC MANNER.

2796
02:03:51,168 --> 02:03:52,837
THERE'S THOUSANDS OF CELL

2797
02:03:52,837 --> 02:03:53,271
SPECIFIC ENHANCERS.

2798
02:03:53,271 --> 02:03:55,806
MUCH OF THE GENOME IS

2799
02:03:55,806 --> 02:03:58,075
TRANSCRIBED INCLUDING LONG

2800
02:03:58,075 --> 02:04:01,245
NON-CODING RNAs, AND MUCH OF

2801
02:04:01,245 --> 02:04:02,680
THE LINKS TO HUMAN DISEASE IS

2802
02:04:02,680 --> 02:04:04,348
NOT IN THE CODING PORTIONS OF

2803
02:04:04,348 --> 02:04:07,785
THE GENE BUT IN THE NON-CODING 

2804
02:04:07,785 --> 02:04:14,825
PORTION OF THE GENOME, ET

2805
02:04:14,825 --> 02:04:15,493
CETERA.

2806
02:04:15,493 --> 02:04:19,664
THE OTHER PART WITH THESE ACTIVE

2807
02:04:19,664 --> 02:04:21,432
ENHANCERS WE HAVE BETTER

2808
02:04:21,432 --> 02:04:27,772
UNDERSTANDING OF THE LOOPING OF 

2809
02:04:27,772 --> 02:04:30,474
THE GENOME, AGAIN CELL SPECIFIC

2810
02:04:30,474 --> 02:04:31,108
INACTIVATION DEPENDENT MATTERS,

2811
02:04:31,108 --> 02:04:33,611
I'LL COME TO THAT AS WELL.

2812
02:04:33,611 --> 02:04:35,246
ONE OF THE STRIKING FINDINGS,

2813
02:04:35,246 --> 02:04:39,684
THIS IS THE WORK FROM DAN

2814
02:04:39,684 --> 02:04:40,551
KASTNER AND MY BOSS LINDSEY

2815
02:04:40,551 --> 02:04:43,087
CRISWELL WE KNOW A LOT ABOUT OF

2816
02:04:43,087 --> 02:04:47,024
ROLE OF MHC AS A LINK IN

2817
02:04:47,024 --> 02:04:48,759
DISEASES, AUTOIMMUNE DISEASE,

2818
02:04:48,759 --> 02:04:53,264
BUT UP HERE IS STAT 4, STRIKING

2819
02:04:53,264 --> 02:04:54,498
TO US.

2820
02:04:54,498 --> 02:04:59,270
WE WORKED A LOT WITH STAT4 OVER

2821
02:04:59,270 --> 02:05:00,204
THE YEARS, DIDN'T EXPECT

2822
02:05:00,204 --> 02:05:02,440
NECESSARILY THAT IT WOULD BE

2823
02:05:02,440 --> 02:05:08,312
ASSOCIATED WITH RHEUMATOID

2824
02:05:08,312 --> 02:05:10,381
ARTHRITIS, LUPUS, ET CETERA.

2825
02:05:10,381 --> 02:05:15,486
THE PRESENCE OF THE STAT4 RISK

2826
02:05:15,486 --> 02:05:16,787
ALLELE, THIS VARIANT IS

2827
02:05:16,787 --> 02:05:17,988
ASSOCIATED WITH MORE SEVERE

2828
02:05:17,988 --> 02:05:18,222
DISEASE.

2829
02:05:18,222 --> 02:05:20,758
WE WERE THINKING IN TERMS OF

2830
02:05:20,758 --> 02:05:25,496
USING JAK INHIBITORS, MIGHT WE

2831
02:05:25,496 --> 02:05:27,198
TAKE THIS INTO CONSIDERATION,

2832
02:05:27,198 --> 02:05:28,232
PATIENTS WITH STAT4 RISK ALLELE

2833
02:05:28,232 --> 02:05:32,970
AND THOSE WHO DIDN'T. 

2834
02:05:32,970 --> 02:05:35,306
I HAVE TO SAY THAT THIS WAS

2835
02:05:35,306 --> 02:05:37,041
BETTY DIAMOND'S IDEA.

2836
02:05:37,041 --> 02:05:39,810
AND WE HAD A CLINICAL TRIAL HERE

2837
02:05:39,810 --> 02:05:43,647
IN THE CLINICAL CENTER, VERY

2838
02:05:43,647 --> 02:05:46,684
SMALL TRIAL WITH 30-SOME-ODD

2839
02:05:46,684 --> 02:05:46,951
SUBJECTS.

2840
02:05:46,951 --> 02:05:48,853
AND I SAID AT THE TIME TO BETTY

2841
02:05:48,853 --> 02:05:51,222
THAT, YOU KNOW, WE'RE NEVER

2842
02:05:51,222 --> 02:05:54,658
GOING TO GET THE SENSITIVITY,

2843
02:05:54,658 --> 02:05:56,160
WE'RE NEVER GOING TO GET IT WITH

2844
02:05:56,160 --> 02:05:57,628
THAT FEW PATIENTS.

2845
02:05:57,628 --> 02:05:58,829
MAYBE IF WE HAD A THOUSAND

2846
02:05:58,829 --> 02:06:01,198
PATIENTS WE COULD SEE THAT.

2847
02:06:01,198 --> 02:06:02,566
THEY DIDN'T LISTEN TO ME IN

2848
02:06:02,566 --> 02:06:10,007
PLANNING THIS, THAT WAS REALLY

2849
02:06:10,007 --> 02:06:14,478
GOOD.

2850
02:06:14,478 --> 02:06:16,781
BECAUSE THEY DEVISED A PROTOCOL,

2851
02:06:16,781 --> 02:06:19,116
MARIANA INTERESTED IN THE ROLE

2852
02:06:19,116 --> 02:06:20,451
OF NEUTROPHILS IN LUPUS, ONE

2853
02:06:20,451 --> 02:06:24,255
THING THAT WAS CLEAR YOU COULD

2854
02:06:24,255 --> 02:06:29,994
SEE THAT THE EFFECT OF JAK

2855
02:06:29,994 --> 02:06:32,663
INHIBITORS AND LEVEL OF

2856
02:06:32,663 --> 02:06:36,233
NETosis WAS MORE PROFOUND WITH

2857
02:06:36,233 --> 02:06:39,703
STAT4 RISK ALLELE, BETTER 

2858
02:06:39,703 --> 02:06:41,772
RESPONSE WITH JAK INHIBITORS.

2859
02:06:41,772 --> 02:06:49,647
SO GOING FURTHER ON THIS I GUESS

2860
02:06:49,647 --> 02:06:51,849
I WASN'T -- HAD LOOKED NOT JUST

2861
02:06:51,849 --> 02:06:53,017
FOR PATIENTS WITH LOSS OF

2862
02:06:53,017 --> 02:06:54,485
FUNCTION OF DIFFERENT STAT

2863
02:06:54,485 --> 02:06:56,854
MOLECULES AND LOSS OF FUNCTION

2864
02:06:56,854 --> 02:06:58,823
OF STATS, ET CETERA, ET CETERA,

2865
02:06:58,823 --> 02:07:00,257
BUT THIS DISEASE REALLY WAS

2866
02:07:00,257 --> 02:07:03,694
REALLY QUITE STRIKING TO ME.

2867
02:07:03,694 --> 02:07:05,496
AND REVEALS A COUPLE LESSONS I

2868
02:07:05,496 --> 02:07:05,863
THINK.

2869
02:07:05,863 --> 02:07:11,602
AND SO THIS IS A DISEASE CALLED

2870
02:07:11,602 --> 02:07:14,004
PAN SCLEROTIC MORPHEA, PATIENTS

2871
02:07:14,004 --> 02:07:18,042
HAVE THIS DEFORMING SKIN DISEASE

2872
02:07:18,042 --> 02:07:19,376
SHOWN HERE IN THIS PATIENT,

2873
02:07:19,376 --> 02:07:21,912
REALLY TRAGIC CHILD WHO HAS

2874
02:07:21,912 --> 02:07:27,785
ACTUALLY THREE AMPUTATION OF

2875
02:07:27,785 --> 02:07:29,119
LIMBS WITH INFLAMMATION IN THE

2876
02:07:29,119 --> 02:07:32,089
SKIN, REALLY AS YOU MIGHT

2877
02:07:32,089 --> 02:07:33,958
IMAGINE WAS REFRACTORY TO

2878
02:07:33,958 --> 02:07:35,226
VARIOUS THERAPIES.

2879
02:07:35,226 --> 02:07:38,529
AND WHAT HAD TURNED OUT, THIS

2880
02:07:38,529 --> 02:07:42,666
WAS A COLLABORATION OF RODERIC

2881
02:07:42,666 --> 02:07:44,568
AND KASTNER AND TOUROC,

2882
02:07:44,568 --> 02:07:48,305
UNIVERSITY OF PITTSBURGH, WHAT

2883
02:07:48,305 --> 02:07:51,775
THEY HAD WAS VARIANTS IN THE SH

2884
02:07:51,775 --> 02:07:53,944
2 DOMAIN, THE STRUCTURE OF

2885
02:07:53,944 --> 02:07:58,415
STATS, DNA COMING OUT AT YOU,

2886
02:07:58,415 --> 02:08:02,486
AND STAT MOLECULES BIND AROUND

2887
02:08:02,486 --> 02:08:05,723
THE DNA, IT'S THIS SH 2 DOMAIN,

2888
02:08:05,723 --> 02:08:07,925
THIS INTERACTION IS THE THING

2889
02:08:07,925 --> 02:08:10,194
THAT PROVIDES THE HINGE OVER

2890
02:08:10,194 --> 02:08:11,529
THESE STAT MOLECULES SHOWN HERE

2891
02:08:11,529 --> 02:08:15,833
AND THESE PATIENTS WHO HAVE GAIN

2892
02:08:15,833 --> 02:08:17,635
OF FUNCTION ALLELES HAVE

2893
02:08:17,635 --> 02:08:18,469
EXAGGERATED STAT ACTIVITY WHICH

2894
02:08:18,469 --> 02:08:21,038
I'LL SHOW IN A MOMENT AND SO

2895
02:08:21,038 --> 02:08:26,911
THAT THIS DISEASE HAS

2896
02:08:26,911 --> 02:08:31,248
EXAGGERATED RESPONSES TO 

2897
02:08:31,248 --> 02:08:33,083
CYTOKINES THAT USE STAT4.

2898
02:08:33,083 --> 02:08:35,986
WE FOUND THAT JAK INHIBITORS ARE

2899
02:08:35,986 --> 02:08:41,325
EFFECTIVE IN THIS SETTING, THAT

2900
02:08:41,325 --> 02:08:43,093
THESE STAT4 VARIANT -- I'M GOING

2901
02:08:43,093 --> 02:08:45,396
TO GET INTO MORE DETAIL IN A

2902
02:08:45,396 --> 02:08:48,699
MOMENT BUT WE ASSOCIATED STAT4

2903
02:08:48,699 --> 02:08:50,901
WITH FUNCTIONING IN IMMUNE

2904
02:08:50,901 --> 02:09:01,612
CELLS, LYMPHOCYTES, ET CETERA, 

2905
02:09:01,612 --> 02:09:02,980
NK CELLS.

2906
02:09:02,980 --> 02:09:05,516
IL-6 CAN SIGNAL THROUGH STAT4

2907
02:09:05,516 --> 02:09:07,117
AND ACTUALLY THEN GIVES A

2908
02:09:07,117 --> 02:09:09,453
FEEDBACK SO THERE'S MORE IL-6

2909
02:09:09,453 --> 02:09:11,689
PRODUCTION AND SO YOU HAVE

2910
02:09:11,689 --> 02:09:14,058
ABERRANT FUNCTION OF FIBROBLAST

2911
02:09:14,058 --> 02:09:15,726
AND INCREASE FIBROSIS AND

2912
02:09:15,726 --> 02:09:17,227
IMPAIRED WOUND HEALING.

2913
02:09:17,227 --> 02:09:18,162
THANKFULLY WITH THESE PATIENTS

2914
02:09:18,162 --> 02:09:20,764
WE NOW HAVE THE OPPORTUNITY TO

2915
02:09:20,764 --> 02:09:26,470
TREAT THESE PATIENTS WITH JAK

2916
02:09:26,470 --> 02:09:26,770
INHIBITORS.

2917
02:09:26,770 --> 02:09:28,572
TURNING TO ANOTHER PART OF THE

2918
02:09:28,572 --> 02:09:33,377
STORY, THIS IS WORK BY RACHEL

2919
02:09:33,377 --> 02:09:36,113
PHILLIPS IN THE LAB, STARTS WITH

2920
02:09:36,113 --> 02:09:38,482
CHRIS BYRON, THIS IS WHAT I WAS

2921
02:09:38,482 --> 02:09:42,186
REFERRING TO BEFORE WHEN WE

2922
02:09:42,186 --> 02:09:44,822
FIRST STUDIED JAK/STAT PATH WAY

2923
02:09:44,822 --> 02:09:48,325
HAD A TENDENCY TO TAKE

2924
02:09:48,325 --> 02:09:49,727
CYTOKINES, THIS USES STAT,

2925
02:09:49,727 --> 02:09:54,698
STAT1, IN THIS CASE, SHOWN HERE,

2926
02:09:54,698 --> 02:09:57,001
STAT1, STAT2, HOW INTERFERON

2927
02:09:57,001 --> 02:09:58,969
WORKS, IL-12, WORK WE DID SOME

2928
02:09:58,969 --> 02:10:02,206
TIME AGO WITH CHRIS BYRON IN MY

2929
02:10:02,206 --> 02:10:03,974
LAB, LINKED IL-12 TO STAT4.

2930
02:10:03,974 --> 02:10:07,544
SO WHEN WE WERE THINKING ABOUT

2931
02:10:07,544 --> 02:10:11,415
THE ACTION OF TYPE I

2932
02:10:11,415 --> 02:10:17,287
INTERFERONS, SIGNAL VIA STAT1

2933
02:10:17,287 --> 02:10:19,056
HOMODIMERS AND ISGF 3, BINDS A

2934
02:10:19,056 --> 02:10:23,927
DIFFERENT ELEMENT IN THE GENOME,

2935
02:10:23,927 --> 02:10:26,096
AND STAT4 ACTIVATES -- IS

2936
02:10:26,096 --> 02:10:27,731
ACTIVATED BY IL-12, THE PART

2937
02:10:27,731 --> 02:10:30,968
THAT DRIVES THIS ACTIVATION

2938
02:10:30,968 --> 02:10:32,002
PHENOTYPE, IN PARTICULAR INDUCES

2939
02:10:32,002 --> 02:10:32,736
INTERFERON GAMMA.

2940
02:10:32,736 --> 02:10:35,873
THAT'S THE KEY PART OF THE STORY

2941
02:10:35,873 --> 02:10:42,146
THAT I'LL EXPLAIN AS I GO ALONG

2942
02:10:42,146 --> 02:10:42,346
HERE.

2943
02:10:42,346 --> 02:10:43,747
SO WHAT WE FOUND AT THE TURN OF

2944
02:10:43,747 --> 02:10:46,850
THE CENTURY OR SO LONG BEFORE WE

2945
02:10:46,850 --> 02:10:48,252
REALLY HAD THE TECHNIQUES THEY

2946
02:10:48,252 --> 02:10:50,054
HAVE NOW IN TERMS OF SEQUENCING

2947
02:10:50,054 --> 02:10:54,491
AND THE LIKE, BUT WHAT CHRIS

2948
02:10:54,491 --> 02:10:57,928
BYRON HELPED US UNDERSTAND WAS

2949
02:10:57,928 --> 02:11:00,864
THAT IN LYMPHOCYTES THAT TYPE I

2950
02:11:00,864 --> 02:11:01,999
INTERFERONS ALSO ACTIVATE STAT4,

2951
02:11:01,999 --> 02:11:06,870
THAT MAKES A LOT MORE SENSE

2952
02:11:06,870 --> 02:11:10,507
BECAUSE LUPUS IS AN

2953
02:11:10,507 --> 02:11:12,309
INTERFERONOPATHY, AND THE

2954
02:11:12,309 --> 02:11:13,477
LINKAGE BETWEEN TYPE I

2955
02:11:13,477 --> 02:11:15,646
INTERFERONS AND STAT4 MAKES

2956
02:11:15,646 --> 02:11:17,848
SENSE WITH LINDSEY CRISWELL AND

2957
02:11:17,848 --> 02:11:19,917
DAN KASTNER'S WORK.

2958
02:11:19,917 --> 02:11:23,220
IN LYMPHOCYTES, IN THE BASAL

2959
02:11:23,220 --> 02:11:28,559
STATE, WHEN YOU FIRST HAVE AN 

2960
02:11:28,559 --> 02:11:29,026
INSULT, INDUCTION OF

2961
02:11:29,026 --> 02:11:33,864
INTERFERENCE, TYPE I INTERFERONS

2962
02:11:33,864 --> 02:11:35,866
PREFERENTIALLY ACTIVATE STAT4

2963
02:11:35,866 --> 02:11:37,301
AND DRIVE ACTIVATION PHENOTYPE

2964
02:11:37,301 --> 02:11:39,937
IN INTERFERON GAM A LATER ON

2965
02:11:39,937 --> 02:11:42,973
STAT1 INCREASES AND THEN YOU

2966
02:11:42,973 --> 02:11:45,876
HAVE MORE PREDOMINANTLY THIS

2967
02:11:45,876 --> 02:11:46,510
ANTI-VIRAL RESPONSE.

2968
02:11:46,510 --> 02:11:46,810
OKAY.

2969
02:11:46,810 --> 02:11:51,782
SO WHAT MIGHT THAT MEAN?

2970
02:11:51,782 --> 02:11:53,484
SO, THERE ARE STAT1 LOSS OF

2971
02:11:53,484 --> 02:11:54,818
FUNCTION VARIANTS, STAT1 GAIN OF

2972
02:11:54,818 --> 02:11:55,619
FUNCTION VARIANTS.

2973
02:11:55,619 --> 02:11:58,555
AND YOU KNOW ALL THIS WORK FROM

2974
02:11:58,555 --> 02:11:59,757
STEVE HOLLAND AND COLLEAGUES

2975
02:11:59,757 --> 02:12:01,925
SHOWN HERE, I COULDN'T FIND A

2976
02:12:01,925 --> 02:12:06,130
PICTURE OF ALEX FREEMAN WITH HER

2977
02:12:06,130 --> 02:12:06,630
PARTNER OVER HERE.

2978
02:12:06,630 --> 02:12:09,700
BUT SOMEBODY SHOULD TAKE A

2979
02:12:09,700 --> 02:12:10,033
PICTURE.

2980
02:12:10,033 --> 02:12:14,304
CHARLIE, ARE YOU STILL HERE? 

2981
02:12:14,304 --> 02:12:14,938
MAYBE HE'S NOT HERE.

2982
02:12:14,938 --> 02:12:16,273
>> WE'LL WORK ON IT.

2983
02:12:16,273 --> 02:12:17,841
>> OKAY.

2984
02:12:17,841 --> 02:12:21,211
IN ANY CASE, STEVE AND JOHN, ET

2985
02:12:21,211 --> 02:12:22,846
CETERA, IDENTIFIED PATIENTS WITH

2986
02:12:22,846 --> 02:12:25,315
STAT1 LOSS OF FUNCTION VARIANTS,

2987
02:12:25,315 --> 02:12:26,483
THESE PATIENTS HAVE BACTERIAL

2988
02:12:26,483 --> 02:12:29,086
AND VIRAL INFECTION BUT ALSO

2989
02:12:29,086 --> 02:12:30,454
IDENTIFIED PATIENTS WITH STAT1

2990
02:12:30,454 --> 02:12:32,156
GAIN OF FUNCTION VARIANT.

2991
02:12:32,156 --> 02:12:36,193
WHAT THAT RESULTS IN IS THAT

2992
02:12:36,193 --> 02:12:38,996
THEY HAVE IMPAIRED RESPONSES TO

2993
02:12:38,996 --> 02:12:41,865
FUNGAL INFECTIONS, CANDIDA SHOWN

2994
02:12:41,865 --> 02:12:45,736
HERE, BECAUSE INTERFERON GAMMA

2995
02:12:45,736 --> 02:12:46,737
AND EXAGGERATING INTERFERON

2996
02:12:46,737 --> 02:12:50,474
RESPONSES IMPAIRS THE TYPE 3

2997
02:12:50,474 --> 02:12:55,245
RESPONSE, IL-17 RESPONSES BUT

2998
02:12:55,245 --> 02:12:56,480
INDUCES LUPUS.

2999
02:12:56,480 --> 02:12:59,283
THE PARADOX HERE THOUGH IS THAT

3000
02:12:59,283 --> 02:13:00,450
PATIENTS WHO HAVE STAT1 GAIN OF

3001
02:13:00,450 --> 02:13:04,421
FUNCTIONS AND OTHER PATIENTS WHO

3002
02:13:04,421 --> 02:13:06,056
HAVE INTERFERONOPATHYS INCLUDING

3003
02:13:06,056 --> 02:13:09,393
CHILDREN WITH DOWN SYNDROME AND

3004
02:13:09,393 --> 02:13:11,361
THAT HAVE INTERFERONOPATHYS

3005
02:13:11,361 --> 02:13:13,130
BECAUSE THEY HAVE DUPLICATION OR

3006
02:13:13,130 --> 02:13:22,539
INCREASED EXPRESSION OF

3007
02:13:22,539 --> 02:13:23,373
INTERFERON RECEPTORS,

3008
02:13:23,373 --> 02:13:24,608
EXAGGERATED INTERFERON RESPONSE,

3009
02:13:24,608 --> 02:13:28,545
SHOULD HAVE IN PRINCIPLE

3010
02:13:28,545 --> 02:13:29,179
EXAGGERATED ANTI-VIRAL RESPONSE

3011
02:13:29,179 --> 02:13:31,181
BUT THAT'S NOT HOW IT WORKS.

3012
02:13:31,181 --> 02:13:32,516
THAT'S THE CASE FOR COVID-19.

3013
02:13:32,516 --> 02:13:36,954
SO WE WANTED TO UNDERSTAND THE

3014
02:13:36,954 --> 02:13:37,688
MECHANISM OF THIS.

3015
02:13:37,688 --> 02:13:39,957
AND SO WE MADE A MOUSE, SHOWN

3016
02:13:39,957 --> 02:13:43,327
HERE, WE PUT THE MUTANT ALLELE

3017
02:13:43,327 --> 02:13:44,828
IN SO THAT THE MICE WOULD BE

3018
02:13:44,828 --> 02:13:46,630
LIKE THESE PATIENTS THAT THEY

3019
02:13:46,630 --> 02:13:48,232
WOULD HAVE A ONE GAIN OF

3020
02:13:48,232 --> 02:13:50,534
FUNCTION ALLELE AND ONE NORMAL

3021
02:13:50,534 --> 02:13:51,969
ALLELE, AND THIS IS

3022
02:13:51,969 --> 02:13:57,074
COLLABORATION AGAIN WITH JOSH

3023
02:13:57,074 --> 02:14:05,315
MILNER AND STEVE AND LEE, WE

3024
02:14:05,315 --> 02:14:07,050
INFECT MICE WITH USUAL VARIANTS,

3025
02:14:07,050 --> 02:14:10,120
VIRUSES THAT WE USUALLY THINK

3026
02:14:10,120 --> 02:14:14,758
ABOUT BUT ALSO WITH SARS, I 

3027
02:14:14,758 --> 02:14:16,026
MANAGED TO COVER THAT.

3028
02:14:16,026 --> 02:14:17,294
THE LEGEND HERE.

3029
02:14:17,294 --> 02:14:22,633
SUFFICE IT TO SAY THAT THE MICE

3030
02:14:22,633 --> 02:14:24,935
RECAPITULATE WHAT YOU SEE IN

3031
02:14:24,935 --> 02:14:29,273
PATIENTS, THAT THE MICE IN THIS

3032
02:14:29,273 --> 02:14:31,608
CASE THIS -- THESE MICE EXPRESS

3033
02:14:31,608 --> 02:14:33,343
THE MUTANT ALLELE IN ALL TISSUES

3034
02:14:33,343 --> 02:14:35,612
BUT WE OBVIOUSLY CAN DO THIS IN

3035
02:14:35,612 --> 02:14:37,214
A TISSUE-SPECIFIC MANNER AS

3036
02:14:37,214 --> 02:14:37,447
WELL.

3037
02:14:37,447 --> 02:14:42,519
AND WHAT YOU SEE IS INCREASE

3038
02:14:42,519 --> 02:14:43,153
SUSCEPTIBLABILITY TO VIRAL

3039
02:14:43,153 --> 02:14:44,688
INFECTION, THIS IS SARS-COV-2 AS

3040
02:14:44,688 --> 02:14:45,055
WELL.

3041
02:14:45,055 --> 02:14:49,459
SO WE WANTED TO UNDERSTAND THE

3042
02:14:49,459 --> 02:14:50,460
MECHANISM AND WHAT RACHEL FOUND,

3043
02:14:50,460 --> 02:14:52,296
IN THE INTEREST OF TIME I WON'T

3044
02:14:52,296 --> 02:14:55,432
SHOW ALL THE DATA, THAT THESE

3045
02:14:55,432 --> 02:14:57,067
MICE HAVE CYTOKINE STORM, AGAIN

3046
02:14:57,067 --> 02:14:58,602
AS WE HAVE BEEN HEARING OVER THE

3047
02:14:58,602 --> 02:15:01,104
PAST THREE YEARS OR SO, BUT ONE

3048
02:15:01,104 --> 02:15:03,307
OF THE SURPRISING THINGS IS THAT

3049
02:15:03,307 --> 02:15:04,541
THESE MICE DON'T MAKE INTERFERON

3050
02:15:04,541 --> 02:15:04,875
GAMMA.

3051
02:15:04,875 --> 02:15:08,679
I JUST TOLD YOU ACTUALLY THAT

3052
02:15:08,679 --> 02:15:10,480
THIS IS AN INTERFERONOPATHY, AND

3053
02:15:10,480 --> 02:15:12,282
YOU HAVE INCREASED RESPONSES TO

3054
02:15:12,282 --> 02:15:13,717
INTERFERONS, AND THERE'S A PAPER

3055
02:15:13,717 --> 02:15:16,019
THAT CAME OUT IN SCIENCE

3056
02:15:16,019 --> 02:15:17,854
TRANSLATIONAL MEDICINE BY

3057
02:15:17,854 --> 02:15:19,489
ANOTHER GROUP THAT MADE THESE

3058
02:15:19,489 --> 02:15:23,060
MICE AS WELL, AND THEY FOUND

3059
02:15:23,060 --> 02:15:23,694
THERE'S PATHOLOGY ASSOCIATED

3060
02:15:23,694 --> 02:15:24,962
WITH INTERFERON GAMMA.

3061
02:15:24,962 --> 02:15:27,364
IF THEY KNOCK OUT INTERFERON

3062
02:15:27,364 --> 02:15:29,132
GAMMA RECEPTOR THEY ACTUALLY

3063
02:15:29,132 --> 02:15:32,035
HAVE REDUCTION IN AUTOIMMUNE

3064
02:15:32,035 --> 02:15:32,269
DISEASE.

3065
02:15:32,269 --> 02:15:35,706
TURNS OUT WHAT HAPPENS IN THESE

3066
02:15:35,706 --> 02:15:41,645
MICE IS THAT YOU HAVE IMPAIRED

3067
02:15:41,645 --> 02:15:42,512
STAT4 ACTIVATION.

3068
02:15:42,512 --> 02:15:50,821
TYPE I INTERFERON IS 

3069
02:15:50,821 --> 02:15:51,755
PREFERENTIALLY ACTIVATE STAT4.

3070
02:15:51,755 --> 02:16:02,232
IN THE MICE YOU HAVE REDUCED

3071
02:16:03,433 --> 02:16:04,334
STAT4 PHOSPHORYLATION.

3072
02:16:04,334 --> 02:16:06,303
MAY BE YOU CAN GIVE INTERFERON

3073
02:16:06,303 --> 02:16:10,507
GAMMA BACK?

3074
02:16:10,507 --> 02:16:13,877
THE PATHOLOGY IS DRIVEN BY

3075
02:16:13,877 --> 02:16:14,845
INTERFERON GAMMA.

3076
02:16:14,845 --> 02:16:16,346
I ALSO WAS THINKING IT'S GOING

3077
02:16:16,346 --> 02:16:18,882
TO BE REALLY HARD TO FIGURE OUT

3078
02:16:18,882 --> 02:16:20,417
WHAT'S THE DOSE OF INTERFERON

3079
02:16:20,417 --> 02:16:23,153
GAMMA, THE TIMING, ET CETERA, ET

3080
02:16:23,153 --> 02:16:23,387
CETERA.

3081
02:16:23,387 --> 02:16:25,489
BUT, AGAIN, RACHEL DIDN'T LISTEN

3082
02:16:25,489 --> 02:16:25,689
TO ME.

3083
02:16:25,689 --> 02:16:28,325
AND WHAT SHE DID WAS TO DO A

3084
02:16:28,325 --> 02:16:30,494
COUPLE THINGS HERE, AND WHAT SHE

3085
02:16:30,494 --> 02:16:33,597
DID WAS THIS IS THE NORMAL

3086
02:16:33,597 --> 02:16:36,333
RESPONSE TO INFECTION SHOWN

3087
02:16:36,333 --> 02:16:36,666
HERE.

3088
02:16:36,666 --> 02:16:39,836
AND HERE ARE THE MICE THAT HAVE

3089
02:16:39,836 --> 02:16:42,606
THE STAT1 GAIN OF FUNCTION

3090
02:16:42,606 --> 02:16:43,607
VARIANT.

3091
02:16:43,607 --> 02:16:46,910
AND THEY DID SOMETHING ELSE

3092
02:16:46,910 --> 02:16:48,745
CLEVER, HOW ABOUT IF WE BLOCK

3093
02:16:48,745 --> 02:16:50,947
INTERFERON GAMMA, DOES THAT GIVE

3094
02:16:50,947 --> 02:16:51,982
THE SAME PHENOTYPE?

3095
02:16:51,982 --> 02:16:54,418
AGAIN WE'VE DONE LOTS OF

3096
02:16:54,418 --> 02:16:55,819
DETAILS, LOTS OF SINGLE CELL

3097
02:16:55,819 --> 02:16:56,953
RNAseq, ET CETERA.

3098
02:16:56,953 --> 02:16:58,422
I'M GOING TO SPARE YOU THAT FOR

3099
02:16:58,422 --> 02:16:59,189
THE MOMENT.

3100
02:16:59,189 --> 02:17:03,693
AND WHAT YOU SEE WAS WHEN YOU

3101
02:17:03,693 --> 02:17:05,295
BLOCK INTERFERON GAMMA YOU SET

3102
02:17:05,295 --> 02:17:06,496
THE SAME PHENOTYPE.

3103
02:17:06,496 --> 02:17:12,436
IF I GIVE THE MICE INTERFERON

3104
02:17:12,436 --> 02:17:16,173
GAMMA EXOGENOUS YOU CAN RESOLVE

3105
02:17:16,173 --> 02:17:20,377
THIS CYTOKINE STORM.

3106
02:17:20,377 --> 02:17:21,978
SO, THIS IS PRETTY INTERESTING

3107
02:17:21,978 --> 02:17:23,847
IN A WAY THAT WHEN WE THINK OF

3108
02:17:23,847 --> 02:17:26,049
THE PATIENTS THAT WE'VE SEEN

3109
02:17:26,049 --> 02:17:29,119
WITH CYTOKINE STORM, IF SOME

3110
02:17:29,119 --> 02:17:31,288
PATIENTS, IN THE SETTING, IF

3111
02:17:31,288 --> 02:17:34,124
THEY -- EVEN IF THEY HAD BASAL

3112
02:17:34,124 --> 02:17:38,462
LEVELS OF INTERFERON GAMMA, YOU 

3113
02:17:38,462 --> 02:17:41,631
KNOW, AT HIGHER LEVELS, THAN

3114
02:17:41,631 --> 02:17:42,699
NORMAL SUBJECTS, BUT THEY

3115
02:17:42,699 --> 02:17:45,202
DIDN'T -- WERE NOT ABLE TO SORT

3116
02:17:45,202 --> 02:17:47,204
OF GET THIS BOOST OF INTERFERON

3117
02:17:47,204 --> 02:17:51,942
GAMMA PRODUCTION YOU WOULD NEED

3118
02:17:51,942 --> 02:17:54,344
DURING THE INFECTIONS, THEN

3119
02:17:54,344 --> 02:17:56,480
ACTUALLY YOU COULD TREAT

3120
02:17:56,480 --> 02:17:57,481
PATIENTS WITH INTERFERON GAMMA,

3121
02:17:57,481 --> 02:17:59,216
THAT MIGHT BE RELATIVE FOR

3122
02:17:59,216 --> 02:18:07,257
PATIENTS WITH DOWN SYNDROME AND

3123
02:18:07,257 --> 02:18:08,425
INTERFERONOPATHYS, ET CETERA.

3124
02:18:08,425 --> 02:18:16,533
SIGNALING IS NOT JUST ABOUT THIS

3125
02:18:16,533 --> 02:18:17,667
ON/OFF SWITCH.

3126
02:18:17,667 --> 02:18:19,202
ONE PART, TO END QUICKLY, ONE

3127
02:18:19,202 --> 02:18:23,173
THING WE LEARNED IN TERMS OF

3128
02:18:23,173 --> 02:18:26,510
THINKING ABOUT CYTOKINES IS THE

3129
02:18:26,510 --> 02:18:27,744
COMPLEXITY OF CYTOKINE GENES

3130
02:18:27,744 --> 02:18:31,114
SHOWN HERE, SO HERE YOU HAVE

3131
02:18:31,114 --> 02:18:32,816
INTERFERON GAMMA, A TINY

3132
02:18:32,816 --> 02:18:33,583
INTERFERON GAMMA GENE, LITTERED

3133
02:18:33,583 --> 02:18:37,354
WITH ALL OF THESE ACTIVE

3134
02:18:37,354 --> 02:18:42,492
ENHANCERS, AND INCLUDES THINGS 

3135
02:18:42,492 --> 02:18:46,163
LIKE LONG ENCODING RNAs,

3136
02:18:46,163 --> 02:18:47,697
INCLUDING INTERFERON GAMMA 1,

3137
02:18:47,697 --> 02:18:50,367
Th1 CELLS WITH ACTIVE

3138
02:18:50,367 --> 02:18:55,405
ENHANCERS, Th2 CELLS WHICH

3139
02:18:55,405 --> 02:18:57,607
DON'T, AND WHAT WE FOUND A FEW

3140
02:18:57,607 --> 02:19:01,845
YEARS AGO THAT IF YOU LOOKED AT

3141
02:19:01,845 --> 02:19:03,480
SUPERENHANCERS IN IMMUNE CELLS,

3142
02:19:03,480 --> 02:19:13,957
THEY OFTEN ENCOMPASS CYTOKINES,

3143
02:19:13,957 --> 02:19:14,524
CYTOKINE RECEPTORS, ANOTHER

3144
02:19:14,524 --> 02:19:15,659
STORY MORRISON PRESENTED THIS

3145
02:19:15,659 --> 02:19:18,061
TWO DAYS AGO BUT I'LL SPARE YOU

3146
02:19:18,061 --> 02:19:20,330
THAT AT THE MOMENT.

3147
02:19:20,330 --> 02:19:23,033
INTERFERE GAMMA AS 1 IS LINKED

3148
02:19:23,033 --> 02:19:24,301
TO IBD.

3149
02:19:24,301 --> 02:19:25,635
THIS IS ANOTHER EXAMPLE OF WHAT

3150
02:19:25,635 --> 02:19:27,571
I WAS SAYING BEFORE THAT THE

3151
02:19:27,571 --> 02:19:30,473
GENETIC LINK IS OUT HERE, NOT

3152
02:19:30,473 --> 02:19:31,775
ACTUALLY IN THE CODING REGION.

3153
02:19:31,775 --> 02:19:36,046
AND WHAT WE DID WAS WE MADE TWO

3154
02:19:36,046 --> 02:19:38,782
MODELS OF INTERFERON GAMMA AS 1

3155
02:19:38,782 --> 02:19:43,954
KNOCKOUT MICE AND SHOWED THAT

3156
02:19:43,954 --> 02:19:47,924
THIS GAVE REDUCED SUSCEPTIBLITY

3157
02:19:47,924 --> 02:19:51,428
OR INCREASED SUSCEPTIBILITY.

3158
02:19:51,428 --> 02:19:53,496
ONE THING, A COLLABORATION WITH

3159
02:19:53,496 --> 02:19:58,335
AHMED, HE FOUND IN  HIS COHORT

3160
02:19:58,335 --> 02:20:00,904
VACCINATED WITH YELLOW FEVER,

3161
02:20:00,904 --> 02:20:02,005
LOOKING AT ANTIGEN-SPECIFIC

3162
02:20:02,005 --> 02:20:06,209
MEMORY T CELLS, HIGHER LEVELS OF

3163
02:20:06,209 --> 02:20:08,211
INTERFERON GAMMA AS1 EVEN WITH

3164
02:20:08,211 --> 02:20:13,516
NORMAL LEVELS OF INTERFERON

3165
02:20:13,516 --> 02:20:14,851
GAMMA.

3166
02:20:14,851 --> 02:20:23,793
FINALLY. 

3167
02:20:23,793 --> 02:20:25,829
SO RETURNING TO WHAT I WAS

3168
02:20:25,829 --> 02:20:28,064
MENTIONING BEFORE ABOUT THE

3169
02:20:28,064 --> 02:20:30,066
LOOPING OF CHROMATIN, SO HERE WE

3170
02:20:30,066 --> 02:20:31,134
HAVE THE INTERFERON GAMMA GENE

3171
02:20:31,134 --> 02:20:32,002
SHOWN HERE.

3172
02:20:32,002 --> 02:20:33,837
HERE IS THE LONG NON-CODING RNA

3173
02:20:33,837 --> 02:20:35,171
THAT I MENTIONED.

3174
02:20:35,171 --> 02:20:37,974
THIS IS A CYTOKINE CLUSTER,

3175
02:20:37,974 --> 02:20:45,849
INTERFERON GAMMA IL- 22 AND IN

3176
02:20:45,849 --> 02:20:47,117
HUMANS IL-26.

3177
02:20:47,117 --> 02:20:50,620
HUMAN GENOME IS PLAID. 

3178
02:20:50,620 --> 02:20:56,760
IS THIS PLAIDNESS IMPORTANT?

3179
02:20:56,760 --> 02:21:03,300
AND SO HUNG HU WHO WILL GIVE A 

3180
02:21:03,300 --> 02:21:06,102
TALK OCTOBER 4, CAREFULLY

3181
02:21:06,102 --> 02:21:08,305
ASSESSED FOLDING OF CHROMATIN IN

3182
02:21:08,305 --> 02:21:12,042
THIS LOCUS, WHAT SHE IDENTIFIED

3183
02:21:12,042 --> 02:21:16,279
WAS CTCF BINDING ELEMENT, MINUS

3184
02:21:16,279 --> 02:21:22,218
70 REGION, USING CRISPR/Cas.

3185
02:21:22,218 --> 02:21:30,427
I'LL CUT TO THE CHASE. 

3186
02:21:30,427 --> 02:21:33,296
WHAT WAS FOUND IF YOU ALTER

3187
02:21:33,296 --> 02:21:35,932
CHROMATIN STRUCTURE NK CELLS AND

3188
02:21:35,932 --> 02:21:38,435
MEMORY CD8 T CELLS, THAT'S FINE.

3189
02:21:38,435 --> 02:21:40,170
THEY CAN MAKE INTERFERON GAMMA.

3190
02:21:40,170 --> 02:21:43,540
WHAT HAPPENS IS THAT IN TH1

3191
02:21:43,540 --> 02:21:47,277
CELLS THAT START IN NAIVE STATE

3192
02:21:47,277 --> 02:21:48,044
AND POLARIZE TO Th1 CELLS

3193
02:21:48,044 --> 02:21:52,782
THAT'S WHERE THIS ELEMENT IS

3194
02:21:52,782 --> 02:21:53,383
CRITICAL.

3195
02:21:53,383 --> 02:21:56,152
ARCHITECTURE DOES MATTER IN THIS

3196
02:21:56,152 --> 02:21:56,486
SETTING.

3197
02:21:56,486 --> 02:21:58,621
AND SO THIS COMES BACK TO THIS

3198
02:21:58,621 --> 02:22:00,490
POINT THAT I THINK ONE OF THE

3199
02:22:00,490 --> 02:22:02,592
CHALLENGES FOR THE NEXT 30

3200
02:22:02,592 --> 02:22:05,562
YEARS, THAT WE HAVE BIOLOGICS,

3201
02:22:05,562 --> 02:22:06,763
THAT'S GREAT.

3202
02:22:06,763 --> 02:22:11,201
WE REALLY -- PATIENTS COME INTO

3203
02:22:11,201 --> 02:22:12,802
THE CLINIC, REALLY DO NOT COME

3204
02:22:12,802 --> 02:22:15,705
IN WHEELCHAIRS AS THEY DID WHEN

3205
02:22:15,705 --> 02:22:18,174
I WAS A RESIDENT, ET CETERA.

3206
02:22:18,174 --> 02:22:21,244
SO WE HAVE BIOLOGICS, CAN TREAT

3207
02:22:21,244 --> 02:22:22,712
THESE PATIENTS, AND WE ALSO HAVE

3208
02:22:22,712 --> 02:22:23,713
JAK INHIBITORS AS WELL.

3209
02:22:23,713 --> 02:22:26,750
I WAS THINKING ABOUT THIS, THIS

3210
02:22:26,750 --> 02:22:28,852
IS OUR BAND, FRANCIS COLLINS, ET

3211
02:22:28,852 --> 02:22:29,352
CETERA.

3212
02:22:29,352 --> 02:22:31,254
AND WHAT YOU MAY NOT THINK ABOUT

3213
02:22:31,254 --> 02:22:34,624
AS MUCH AS I THINK ABOUT WHEN

3214
02:22:34,624 --> 02:22:36,226
WE'RE PLAYING IS THAT SOMEBODY

3215
02:22:36,226 --> 02:22:39,262
HAS TO PUT ALL THIS STUFF

3216
02:22:39,262 --> 02:22:41,197
TOGETHER, YOU HAVE THESE MIXING

3217
02:22:41,197 --> 02:22:44,067
THAT'S GOING ON BEHIND THE

3218
02:22:44,067 --> 02:22:45,568
SCENE, SO THAT IT SOUNDS

3219
02:22:45,568 --> 02:22:47,670
BEARABLE TO YOU.

3220
02:22:47,670 --> 02:22:54,110
WHEN I'M OFF KEY OR PLAYING THE

3221
02:22:54,110 --> 02:22:55,912
WRONG SONG, ARRINGTON PRICE OR A

3222
02:22:55,912 --> 02:22:57,414
COLLEAGUE CAN TURN ME DOWN

3223
02:22:57,414 --> 02:22:57,847
QUICKLY.

3224
02:22:57,847 --> 02:23:00,917
I WAS THINKING THIS REMINDS ME

3225
02:23:00,917 --> 02:23:04,954
OF THE INTERFERON GAMMA GENE, WE

3226
02:23:04,954 --> 02:23:07,424
HAVE ALL THESE SWITCHES, JUST

3227
02:23:07,424 --> 02:23:08,124
SURROUNDING THIS REGION.

3228
02:23:08,124 --> 02:23:11,761
AND THE ISSUE OF COURSE IS DO WE

3229
02:23:11,761 --> 02:23:13,229
TAKE THAT INTO CONSIDERATION AS

3230
02:23:13,229 --> 02:23:15,999
WE'RE SORT OF THINKING ABOUT

3231
02:23:15,999 --> 02:23:16,766
TREATMENT?

3232
02:23:16,766 --> 02:23:20,003
YOU KNOW, WE HAVE BIOLOGICS, JAK

3233
02:23:20,003 --> 02:23:24,374
INHIBITORS, WHAT WE NEED TO

3234
02:23:24,374 --> 02:23:26,543
UNDERSTAND IS MORE SOPHISTICATED

3235
02:23:26,543 --> 02:23:28,344
BIOMARKERS IN PATIENTS,

3236
02:23:28,344 --> 02:23:29,379
CONSEQUENCE OF CYTOKINE

3237
02:23:29,379 --> 02:23:30,146
SIGNALING, ET CETERA.

3238
02:23:30,146 --> 02:23:31,714
AND REALLY UNDERSTAND AS I WAS

3239
02:23:31,714 --> 02:23:33,583
REFERRING TO BEFORE WITH THE

3240
02:23:33,583 --> 02:23:37,720
STAT 4 RISK ALLELE AND PATIENTS

3241
02:23:37,720 --> 02:23:38,888
HAVE VARIOUS GAIN OF FUNCTION

3242
02:23:38,888 --> 02:23:40,056
MUTATIONS TO UNDERSTAND THAT SO

3243
02:23:40,056 --> 02:23:41,691
WE CAN PICK THE RIGHT DRUGS AT

3244
02:23:41,691 --> 02:23:42,625
THE RIGHT DOSES.

3245
02:23:42,625 --> 02:23:43,927
REALLY ULTIMATELY I THINK WE

3246
02:23:43,927 --> 02:23:48,631
NEED TO UNDERSTAND THE

3247
02:23:48,631 --> 02:23:49,999
COMPLEXITY OF GENE REGULATION

3248
02:23:49,999 --> 02:23:51,501
THERAPEUTICALLY AND REALLY IN

3249
02:23:51,501 --> 02:23:54,204
MOST DISEASES EVERYTHING FROM

3250
02:23:54,204 --> 02:23:56,539
HYPERTENSION TO CANCER, IT'S

3251
02:23:56,539 --> 02:23:57,407
COMBINATORIAL THERAPY.

3252
02:23:57,407 --> 02:24:00,610
TYPICALLY YOU GO TO THE DOCTOR,

3253
02:24:00,610 --> 02:24:01,744
WHATEVER, SAY, NO, OKAY, THAT

3254
02:24:01,744 --> 02:24:02,946
DIDN'T WORK, LET ME GIVE YOU

3255
02:24:02,946 --> 02:24:04,714
THIS OTHER DRUG.

3256
02:24:04,714 --> 02:24:06,516
IT'S NOT RATIONALE, DOESN'T TAKE

3257
02:24:06,516 --> 02:24:08,318
INTO ACCOUNT ALL OF THE FACTORS

3258
02:24:08,318 --> 02:24:09,953
REALLY THAT REGULATE ALL THESE

3259
02:24:09,953 --> 02:24:10,854
KEY GENES.

3260
02:24:10,854 --> 02:24:13,122
I WOULD ARGUE THAT THIS IS JUST

3261
02:24:13,122 --> 02:24:14,491
THE BEGINNING OF OUR

3262
02:24:14,491 --> 02:24:16,059
UNDERSTANDING OF IMMUNE CELL

3263
02:24:16,059 --> 02:24:16,326
SIGNALING.

3264
02:24:16,326 --> 02:24:18,561
AND I THINK I MENTIONED PEOPLE'S

3265
02:24:18,561 --> 02:24:22,065
NAMES ALONG THE WAY.

3266
02:24:22,065 --> 02:24:22,999
AGAIN, JUST TREMENDOUSLY

3267
02:24:22,999 --> 02:24:25,368
THANKFUL JUST TO BE HERE AT THE

3268
02:24:25,368 --> 02:24:25,535
NIH.

3269
02:24:25,535 --> 02:24:26,503
SO THANKS.

3270
02:24:26,503 --> 02:24:28,004
[APPLAUSE]

3271
02:24:28,004 --> 02:24:34,010
 

3272
02:24:34,010 --> 02:24:34,477
 

3273
02:24:34,477 --> 02:24:37,113
>> DR. O'SHEA'S BAND IS
PLAYING

3274
02:24:37,113 --> 02:24:41,117
TOMORROW AT THE CLOSE.

3275
02:24:41,117 --> 02:24:41,384
>> NOT ME.

3276
02:24:41,384 --> 02:24:42,652
>> WE GOT A SOUND PERSON.

3277
02:24:42,652 --> 02:24:44,554
YOU CAN ADD ME TO YOUR

3278
02:24:44,554 --> 02:24:45,088
ACKNOWLEDGMENT PAGE.

3279
02:24:45,088 --> 02:24:46,956
>> I HAVE TO MAKE A
COMMENT.

3280
02:24:46,956 --> 02:24:49,359
I HAVE TO SAY FIRST OF ALL I'VE

3281
02:24:49,359 --> 02:24:54,430
ALWAYS THOUGHT IT IS INDICATIVE

3282
02:24:54,430 --> 02:24:57,000
OF INCREDIBLE HEALTHY EGO OF THE

3283
02:24:57,000 --> 02:24:59,102
SCIENTIFIC COMMUNITY THAT

3284
02:24:59,102 --> 02:25:00,336
BECAUSE WE DON'T UNDERSTAND 98%

3285
02:25:00,336 --> 02:25:03,473
OF THE GENOME WE JUST CALL IT

3286
02:25:03,473 --> 02:25:04,240
JUNK, CLEARLY NOT.

3287
02:25:04,240 --> 02:25:06,109
AND THE OTHER THING THAT

3288
02:25:06,109 --> 02:25:08,344
OCCURRED TO ME, IT'S ALWAYS BEEN

3289
02:25:08,344 --> 02:25:10,780
AMAZING TO ME, SINCE EARLY IN

3290
02:25:10,780 --> 02:25:12,482
MEDICAL SCHOOL, THAT THE SAME

3291
02:25:12,482 --> 02:25:16,686
PERSON THAT MAKES SUCH A ROBUST

3292
02:25:16,686 --> 02:25:20,390
IMMUNE RESPONSE THAT THEY

3293
02:25:20,390 --> 02:25:22,125
DESTROY THEIR GENES, THEIR

3294
02:25:22,125 --> 02:25:25,929
JOINTS, THEIR G.I. TRACT, THEIR

3295
02:25:25,929 --> 02:25:28,364
HEART, CAN'T MAKE AN IMMUNE

3296
02:25:28,364 --> 02:25:30,466
RESPONSE TO KILL OFF THE BUGS

3297
02:25:30,466 --> 02:25:32,635
THAT INFECT ALL OF US, I MEAN,

3298
02:25:32,635 --> 02:25:36,005
THAT IS -- THAT IN ITSELF SAYS

3299
02:25:36,005 --> 02:25:40,310
THIS IS A REGULATORY DISORDER,

3300
02:25:40,310 --> 02:25:45,815
AND NOT AN IMMUNE SINGLE

3301
02:25:45,815 --> 02:25:46,849
FUNCTION DISORDER.

3302
02:25:46,849 --> 02:25:47,650
ANYWAY, VERY INTERESTING.

3303
02:25:47,650 --> 02:25:50,620
I THINK YOU SEE THE THREAD IN

3304
02:25:50,620 --> 02:25:53,690
THIS THAT EVERY ANSWER TO A

3305
02:25:53,690 --> 02:25:56,025
QUESTION GENERATES 100 MORE

3306
02:25:56,025 --> 02:25:56,326
QUESTIONS.

3307
02:25:56,326 --> 02:25:58,461
AND THAT IS THE JOY OF WHAT WE

3308
02:25:58,461 --> 02:26:00,730
ALL DO.

3309
02:26:00,730 --> 02:26:02,198
SO OUR FINAL SPEAKER THIS

3310
02:26:02,198 --> 02:26:10,106
AFTERNOON IS DR. DEBORAH

3311
02:26:10,106 --> 02:26:13,676
MORRISON, SENIOR INVESTIGATOR AT

3312
02:26:13,676 --> 02:26:16,312
NCI CCR, AN EXPERT IN THE

3313
02:26:16,312 --> 02:26:20,917
FUNCTION OF THE RAS PROTEIN

3314
02:26:20,917 --> 02:26:23,419
KINASE PATHWAYS, AND SHE HAS

3315
02:26:23,419 --> 02:26:25,388
STUDIED BOTH THE NORMAL FUNCTION

3316
02:26:25,388 --> 02:26:28,291
OF THOSE PATHWAYS AND THE WAY

3317
02:26:28,291 --> 02:26:31,594
THOSE PATHWAYS AND THEIR

3318
02:26:31,594 --> 02:26:33,429
ABERRATIONS LEAD TO DISEASE.

3319
02:26:33,429 --> 02:26:37,200
AND, AGAIN, TO LOOK AT THE

3320
02:26:37,200 --> 02:26:38,101
BIOCHEMICAL AND STRUCTURAL

3321
02:26:38,101 --> 02:26:41,671
ASPECTS OF RAS AND THE PROTEINS

3322
02:26:41,671 --> 02:26:44,073
WITH WHICH IT INTERACTS TO TRY

3323
02:26:44,073 --> 02:26:46,409
AND DEVELOP NEW THERAPEUTIC

3324
02:26:46,409 --> 02:26:46,709
STRATEGIES.

3325
02:26:46,709 --> 02:26:47,110
DR. MORRISON.

3326
02:26:47,110 --> 02:26:50,513
[APPLAUSE]

3327
02:26:50,513 --> 02:26:57,420
 

3328
02:26:57,420 --> 02:26:57,553
 

3329
02:26:57,553 --> 02:27:02,125
>> IT'S GREAT TO BE HERE
TODAY.

3330
02:27:02,125 --> 02:27:04,060
AND I WANT TO GIVE YOU -- TO

3331
02:27:04,060 --> 02:27:06,496
GIVE ME A CHANCE TO TELL YOU

3332
02:27:06,496 --> 02:27:09,165
ABOUT MY JOURNEY IN SCIENCE.

3333
02:27:09,165 --> 02:27:09,899
LET'S SEE.

3334
02:27:09,899 --> 02:27:14,771
I WANT TO GO FORWARD.

3335
02:27:14,771 --> 02:27:24,547
 

3336
02:27:24,547 --> 02:27:25,081
 

3337
02:27:25,081 --> 02:27:26,115
I NEED HELP.

3338
02:27:26,115 --> 02:27:33,823
JUST TO GET ME STARTED. 

3339
02:27:33,823 --> 02:27:36,192
SO, ONE OF THE MAJOR FOCUSES OF

3340
02:27:36,192 --> 02:27:43,866
MY LAB HAS BEEN IN THE STUDY OF

3341
02:27:43,866 --> 02:27:47,804
SINGLE -- SIGNAL TRANSDUCTION

3342
02:27:47,804 --> 02:27:52,208
PATHWAY REGULATED BY RAF FAMILY

3343
02:27:52,208 --> 02:27:52,975
OF GTPaseS.

3344
02:27:52,975 --> 02:27:57,313
LET ME GO BACK.

3345
02:27:57,313 --> 02:27:57,447
 

3346
02:27:57,447 --> 02:27:58,881
CLEARLY I'VE NEVER USED THIS

3347
02:27:58,881 --> 02:28:01,050
POINTER BEFORE.

3348
02:28:01,050 --> 02:28:03,486
 

3349
02:28:03,486 --> 02:28:12,395
I WANT TO GO BACK. 

3350
02:28:12,395 --> 02:28:13,062
BOTTOM?

3351
02:28:13,062 --> 02:28:21,738
NO, THIS IS GOING FORWARD. 

3352
02:28:21,738 --> 02:28:31,781
 

3353
02:28:49,665 --> 02:28:50,299
>> THERE YOU GO.

3354
02:28:50,299 --> 02:28:51,734
>> ANYHOW, WE'RE
INTERESTED IN

3355
02:28:51,734 --> 02:28:56,038
THE PATHWAY THAT'S REGULATED BY

3356
02:28:56,038 --> 02:28:58,674
THE RAF GTPase, AND THIS PLAY

3357
02:28:58,674 --> 02:29:01,144
AS VERY IMPORTANT ROLE IN

3358
02:29:01,144 --> 02:29:02,445
TRANSMISSION OF MANY CELLULAR

3359
02:29:02,445 --> 02:29:05,381
SIGNALS AND OFTEN SIGNAL IS

3360
02:29:05,381 --> 02:29:07,416
INITIATED BY EITHER GROWTH

3361
02:29:07,416 --> 02:29:10,219
FACTOR OR LIGAND BINDING TO

3362
02:29:10,219 --> 02:29:12,655
RECEPTOR TYROSINE KINASE ON THE

3363
02:29:12,655 --> 02:29:23,132
CELL SURFACE THAT LEADS GTP

3364
02:29:23,666 --> 02:29:26,202
LOADING, A CRITICAL FACTOR

3365
02:29:26,202 --> 02:29:30,139
COMPONENTS OF CASCADE, AND A

3366
02:29:30,139 --> 02:29:32,942
PROTEIN KINASE, THE RAF KINASES.

3367
02:29:32,942 --> 02:29:36,546
I AGAIN WORKING ON RAS AS A

3368
02:29:36,546 --> 02:29:41,184
FIRST-YEAR POSTDOC IN IT TOM

3369
02:29:41,184 --> 02:29:43,152
ROBERTS' LAB AT DANA-FARBER

3370
02:29:43,152 --> 02:29:44,754
CANCER INSTITUTE, 30 YEARS LATER

3371
02:29:44,754 --> 02:29:46,055
I'M STILL WORKING ON IT.

3372
02:29:46,055 --> 02:29:48,758
BE CAREFUL WHAT YOU CHOOSE AS A

3373
02:29:48,758 --> 02:29:54,330
POSTDOC TO STUDY.

3374
02:29:54,330 --> 02:29:55,131
SO, LET'S SEE.

3375
02:29:55,131 --> 02:29:56,866
IN ADDITION TO PLAYING AN

3376
02:29:56,866 --> 02:29:58,734
IMPORTANT ROLE IN NORMAL GROWTH

3377
02:29:58,734 --> 02:30:00,536
AND DEVELOPMENT, WE ALSO KNOW

3378
02:30:00,536 --> 02:30:02,872
THIS PATHWAY PLAYS A VERY

3379
02:30:02,872 --> 02:30:04,574
IMPORTANT ROLE IN HUMAN DISEASE

3380
02:30:04,574 --> 02:30:06,409
STATES AS WELL.

3381
02:30:06,409 --> 02:30:11,080
AND SOMATIC MUTATIONS IN VARIOUS

3382
02:30:11,080 --> 02:30:13,149
COMPONENTS OF THE PATHWAY ARE

3383
02:30:13,149 --> 02:30:18,988
KNOWN TO BE DRIVERS IN CERTAIN 

3384
02:30:18,988 --> 02:30:19,589
CANCERS.

3385
02:30:19,589 --> 02:30:21,257
IN ADDITION, GERMLINE MUTATIONS

3386
02:30:21,257 --> 02:30:23,759
AND COMPONENTS OF THE PATHWAY

3387
02:30:23,759 --> 02:30:26,863
CAN ALSO BE CAUSATIVE FOR A

3388
02:30:26,863 --> 02:30:27,830
GROUP OF DEVELOPMENTAL

3389
02:30:27,830 --> 02:30:36,072
DISORDERS, SHOWN HERE, KNOWN 

3390
02:30:36,072 --> 02:30:40,276
COLLECTIVELY AS RASOPATHYS. 

3391
02:30:40,276 --> 02:30:46,282
BECAUSE OF THAT, FOR THE PAST 25

3392
02:30:46,282 --> 02:30:49,151
YEARS EFFORTS TO TARGET

3393
02:30:49,151 --> 02:30:51,320
COMPONENT PATHWAYS FOR

3394
02:30:51,320 --> 02:30:54,156
THERAPEUTIC INTERVENTION.

3395
02:30:54,156 --> 02:30:56,893
BUT I THINK ONE OF THE LESSONS

3396
02:30:56,893 --> 02:30:59,495
WE'VE LEARNED OVER TIME IS THAT

3397
02:30:59,495 --> 02:31:02,131
IT'S REALLY IMPORTANT FOR THIS

3398
02:31:02,131 --> 02:31:07,036
KIND OF DEVELOPMENT TO BE MOST

3399
02:31:07,036 --> 02:31:08,471
OFFICIAL AND TO WORK THE BEST

3400
02:31:08,471 --> 02:31:10,706
YOU NEED TO UNDERSTAND HOW

3401
02:31:10,706 --> 02:31:13,743
NORMAL SIGNALING THROUGH THE

3402
02:31:13,743 --> 02:31:16,579
PATHWAY TAKES PLACE.

3403
02:31:16,579 --> 02:31:19,715
AND IN -- WHAT HAS TURNED OUT IS

3404
02:31:19,715 --> 02:31:24,954
FOR THE RAF KINASES THEIR 

3405
02:31:24,954 --> 02:31:28,291
REGULATION IS HIGHLY COMPLEX.

3406
02:31:28,291 --> 02:31:33,162
 

3407
02:31:33,162 --> 02:31:35,364
 

3408
02:31:35,364 --> 02:31:37,600
SO FOR THOSE WHO DON'T THINK

3409
02:31:37,600 --> 02:31:41,304
ABOUT THE RAFs ALL DAY EVERY

3410
02:31:41,304 --> 02:31:44,740
DAY, THEY ARE BASICALLY -- THEY

3411
02:31:44,740 --> 02:31:47,877
CONSIST OF C-TERMINAL CATALYTIC

3412
02:31:47,877 --> 02:31:49,178
DOMAIN, N-TERMINAL REGULATORY

3413
02:31:49,178 --> 02:31:54,483
DOMAIN, CONTAINS A RAF BINDING

3414
02:31:54,483 --> 02:31:54,717
DOMAIN.

3415
02:31:54,717 --> 02:31:59,989
WHICH TURNS OUT THERE ARE THREE

3416
02:31:59,989 --> 02:32:03,526
RAF FAMILY MEMBERS ARAF, BRAF, C

3417
02:32:03,526 --> 02:32:06,062
RAF, EVERY ONE CAN FUNCTION AS

3418
02:32:06,062 --> 02:32:07,596
EFFECTOR OF RAF, DIRECTLY BINDS

3419
02:32:07,596 --> 02:32:11,600
RAF, ALL CAN ALSO FUNCTION AS

3420
02:32:11,600 --> 02:32:13,402
INITIATING KINASE IN THE

3421
02:32:13,402 --> 02:32:13,736
CASCADE.

3422
02:32:13,736 --> 02:32:15,604
IF WE THINK ABOUT WHAT'S

3423
02:32:15,604 --> 02:32:16,939
DIFFERENT ABOUT THEM, WELL, IT

3424
02:32:16,939 --> 02:32:20,776
REALLY IS IN TERMS OF ENZYMATIC

3425
02:32:20,776 --> 02:32:21,043
ACTIVITY.

3426
02:32:21,043 --> 02:32:22,445
AND THE INTRINSIC CATALYTIC

3427
02:32:22,445 --> 02:32:24,814
ACTIVITY OF BRAF IS BY FAR THE

3428
02:32:24,814 --> 02:32:25,982
HIGHEST.

3429
02:32:25,982 --> 02:32:28,617
THEN C RAF, THEN ARAF.

3430
02:32:28,617 --> 02:32:31,120
WHAT DISTINGUISHES BRAF IS

3431
02:32:31,120 --> 02:32:32,755
DIFFERENTS WHERE THE ASTERISKS

3432
02:32:32,755 --> 02:32:34,023
ARE, DIFFERENCE IN SEQUENCES AT

3433
02:32:34,023 --> 02:32:37,026
THE BEGINNING OF THE KINASE

3434
02:32:37,026 --> 02:32:39,862
DOMAIN THAT DETERMINE ITS HIGH

3435
02:32:39,862 --> 02:32:42,164
LEVEL OF BASAL KINASE ACTIVITY.

3436
02:32:42,164 --> 02:32:45,234
SO, WORK FROM OUR LAB AND A

3437
02:32:45,234 --> 02:32:46,936
NUMBER OF LABS IN THE FIELD HAVE

3438
02:32:46,936 --> 02:32:49,338
KIND OF LED TO THIS MODEL OF HOW

3439
02:32:49,338 --> 02:32:54,243
THE RAFs ARE REGULATED.

3440
02:32:54,243 --> 02:32:56,512
BRAF AND CRAF ARE PRIMARY

3441
02:32:56,512 --> 02:32:57,313
DISEASE DRIVERS.

3442
02:32:57,313 --> 02:33:00,850
BRAF IS IMPORTANT IN CANCER,

3443
02:33:00,850 --> 02:33:03,619
c-RAF AND BRAF TOGETHER ARE

3444
02:33:03,619 --> 02:33:13,529
DRIVERS IN THE RAFOPATHYS.

3445
02:33:13,529 --> 02:33:15,698
THEY EXIST AS AUTOINHIBITTIVE

3446
02:33:15,698 --> 02:33:19,935
MONOMERS IN THE CYTOSOL AND IT'S

3447
02:33:19,935 --> 02:33:22,671
BOUND TO DIMER OF 14-3-3.

3448
02:33:22,671 --> 02:33:24,173
ONE HIGHLIGHT OF MY CAREER IN

3449
02:33:24,173 --> 02:33:29,478
THE LAST COUPLE YEARS HAS BEEN A

3450
02:33:29,478 --> 02:33:37,620
REALLY WONDERFUL COLLABORATION

3451
02:33:37,620 --> 02:33:45,027
WITH A YOUNG TENURE TRACK 

3452
02:33:45,027 --> 02:33:47,196
INVESTIGATOR OF NCI, ABLE TO

3453
02:33:47,196 --> 02:33:50,499
SOLVE THE CryoEM STRUCTURE,

3454
02:33:50,499 --> 02:33:53,536
YOU CAN SEE THE 14-3-3 DIMER IS

3455
02:33:53,536 --> 02:33:57,807
A CRADLE, NIGH CASE ON TOP.

3456
02:33:57,807 --> 02:34:01,010
RBD WAS ALSO RESOLVED.

3457
02:34:01,010 --> 02:34:07,883
AND THE RBD REPRESENTS THE RAF

3458
02:34:07,883 --> 02:34:10,052
BINDING DOMAIN, A BREAKTHROUGH

3459
02:34:10,052 --> 02:34:12,421
WHEN WE LEARNED THEY WERE DIRECT

3460
02:34:12,421 --> 02:34:14,023
EFFECTORS OF RAF.

3461
02:34:14,023 --> 02:34:18,494
AND THEY CAN DIRECTLY INTERACT

3462
02:34:18,494 --> 02:34:21,464
RAS, FIRST IT RECRUITS THE RAF

3463
02:34:21,464 --> 02:34:23,732
KINASES TO THE PLASMA MEMBRANE,

3464
02:34:23,732 --> 02:34:27,036
PROMOTES THE RELEASE OF THE

3465
02:34:27,036 --> 02:34:29,605
AUTOINHIBITED STATE AND RESULTS

3466
02:34:29,605 --> 02:34:34,643
IN RAF DIMERIZATION WHICH WE

3467
02:34:34,643 --> 02:34:37,780
KNOW IS ESSENTIAL IN MOST

3468
02:34:37,780 --> 02:34:41,383
INSTANCES FOR RAF TO BECOME AN

3469
02:34:41,383 --> 02:34:42,785
ACTIVE KINASE.

3470
02:34:42,785 --> 02:34:45,121
AND WHAT I'D LIKE TO DO IN THE

3471
02:34:45,121 --> 02:34:47,223
REMAINING TIME IS TELL YOU A

3472
02:34:47,223 --> 02:34:48,524
LITTLE BIT ABOUT A RECENT

3473
02:34:48,524 --> 02:34:51,560
PROJECT THAT WE'VE DONE THAT I

3474
02:34:51,560 --> 02:34:52,695
THINK REALLY HIGHLIGHTS SOME OF

3475
02:34:52,695 --> 02:34:53,796
THE COLLABORATIONS THAT WE'VE

3476
02:34:53,796 --> 02:34:56,031
BEEN ABLE TO TAKE ADVANTAGE OF

3477
02:34:56,031 --> 02:34:58,334
HERE AT THE NIH.

3478
02:34:58,334 --> 02:35:01,170
AND ALSO, IT ALLOWED US TO

3479
02:35:01,170 --> 02:35:03,873
DOVETAIL IN AN EXCITING NEW

3480
02:35:03,873 --> 02:35:05,474
INITIATIVE, THAT'S ALSO GOING ON

3481
02:35:05,474 --> 02:35:06,842
AT NIH.

3482
02:35:06,842 --> 02:35:10,146
SO AS I SAID, BRAF IS ONE OF THE

3483
02:35:10,146 --> 02:35:12,581
RAF FAMILY MEMBERS THAT PLAYS A

3484
02:35:12,581 --> 02:35:16,185
ROLE IN HUMAN DISEASE STATE.

3485
02:35:16,185 --> 02:35:18,487
BOTH IN CANCER AND IN

3486
02:35:18,487 --> 02:35:19,088
RASOPATHYS.

3487
02:35:19,088 --> 02:35:21,790
WHAT'S INTERESTING IS THAT THE

3488
02:35:21,790 --> 02:35:23,125
CANCER ASSOCIATED BRAF MUTATIONS

3489
02:35:23,125 --> 02:35:25,427
HAVE BEEN FOUND TO OCCUR

3490
02:35:25,427 --> 02:35:28,697
PRIMARILY IN THE KINASE OR

3491
02:35:28,697 --> 02:35:29,865
CATALYTIC DOMAIN.

3492
02:35:29,865 --> 02:35:31,967
OFTEN STRONG GAIN OF FUNCTION.

3493
02:35:31,967 --> 02:35:37,573
AS A RESULT BECAUSE THEY ARE SO

3494
02:35:37,573 --> 02:35:38,174
ACTIVE, THEIR EMBRYONIC

3495
02:35:38,174 --> 02:35:39,441
(INDISCERNIBLE) IF YOU TRY TO

3496
02:35:39,441 --> 02:35:42,745
MAKE A MODEL.

3497
02:35:42,745 --> 02:35:43,412
RASOPATHY-ASSOCIATED MUTATIONS

3498
02:35:43,412 --> 02:35:45,381
TEND TO BE WEAKER GAIN OF

3499
02:35:45,381 --> 02:35:49,618
FUNCTION, OFTEN OBSERVED IN THE

3500
02:35:49,618 --> 02:35:51,887
N-TERMINAL REGULATORY DOMAIN, IN

3501
02:35:51,887 --> 02:35:55,991
PARTICULAR FOR RASOPATHYS, FOR

3502
02:35:55,991 --> 02:35:57,359
BRAF, CYSTEINE-RICH DOMAIN HAS

3503
02:35:57,359 --> 02:36:01,230
BEEN A UNIQUE HOT SPOT.

3504
02:36:01,230 --> 02:36:03,465
AND FOR THOSE MAYBE NOT SO

3505
02:36:03,465 --> 02:36:05,834
FAMILIAR WITH THE RASOPATHYS,

3506
02:36:05,834 --> 02:36:06,802
IT'S A DEVELOPMENTAL SYNDROME

3507
02:36:06,802 --> 02:36:09,004
AND THERE'S A GROUP OF THEM

3508
02:36:09,004 --> 02:36:09,939
LISTED HERE.

3509
02:36:09,939 --> 02:36:13,042
AND THEY ARE CAUSED BY GERMLINE

3510
02:36:13,042 --> 02:36:17,546
MUTATION IN MEMBERS OF THE RAS

3511
02:36:17,546 --> 02:36:22,918
OR RAS KINASE PATHWAY OCCUR IN 1

3512
02:36:22,918 --> 02:36:30,559
IN A THOUSAND TO 1 IN 2,000 LIVE

3513
02:36:30,559 --> 02:36:40,569
BIRTHS, FOR NOONAN 7% HAVE C RAF

3514
02:36:40,569 --> 02:36:41,604
MUTATION

3515
02:36:41,604 --> 02:36:44,406
70% IN CARDIO FASCIOCUTANEOUS.

3516
02:36:44,406 --> 02:36:49,345
THE RASOPATHYS CAN BE BROADLY

3517
02:36:49,345 --> 02:36:51,513
CHARACTERIZED BY DISTINCT

3518
02:36:51,513 --> 02:36:52,648
CRANIOFACIAL FEATURES,

3519
02:36:52,648 --> 02:36:54,016
CONGENITAL HEART DEFECTS,

3520
02:36:54,016 --> 02:36:57,119
PATIENTS ARE OFTEN SHORT

3521
02:36:57,119 --> 02:36:58,854
STATURE, THEY HAVE LEARNING

3522
02:36:58,854 --> 02:37:03,259
DISABILITIES, AND INCREASED RISK

3523
02:37:03,259 --> 02:37:04,460
OF CERTAIN CHILDHOOD CANCERS.

3524
02:37:04,460 --> 02:37:08,163
SO I SAID THAT FOR BRAF, AGAIN

3525
02:37:08,163 --> 02:37:12,968
WHICH 71% OF PATIENTS WITH CFC

3526
02:37:12,968 --> 02:37:14,870
HAVE BRAF MUTATION, CRD IS A

3527
02:37:14,870 --> 02:37:15,971
PARTICULAR HOT SPOT.

3528
02:37:15,971 --> 02:37:19,508
WE WANTED TO UNDERSTAND WHAT WAS

3529
02:37:19,508 --> 02:37:21,010
THE BASIS FOR THIS.

3530
02:37:21,010 --> 02:37:24,480
AND SO JUST TO LET YOU KNOW WHAT

3531
02:37:24,480 --> 02:37:28,417
A CYSTEINE-RICH DOMAIN IS, IT IS

3532
02:37:28,417 --> 02:37:32,321
A SMALL, 15 AMINO ACID DOMAIN IN

3533
02:37:32,321 --> 02:37:34,323
PROTEINS, COORDINATE ZINC IONS,

3534
02:37:34,323 --> 02:37:36,358
TWO TYPES FOUND.

3535
02:37:36,358 --> 02:37:39,428
TYPICAL AND ATYPICAL.

3536
02:37:39,428 --> 02:37:42,131
AND A CLASSICAL TYPICAL C1

3537
02:37:42,131 --> 02:37:47,770
DOMAIN IS CONTAINED IN PROTEIN

3538
02:37:47,770 --> 02:37:51,106
KINASE C, BINDS.

3539
02:37:51,106 --> 02:37:53,475
ATYPICAL ONES, ATYPICAL IS THAT

3540
02:37:53,475 --> 02:37:55,544
THERE'S A TRUNCATION IN LOOP 2,

3541
02:37:55,544 --> 02:38:01,083
WHICH DOESN'T ALLOW THEM TO

3542
02:38:01,083 --> 02:38:02,718
INTERACT, BUT THE ATYPICAL

3543
02:38:02,718 --> 02:38:05,421
CRDs ARE OFTEN FOUND IN

3544
02:38:05,421 --> 02:38:06,422
SIGNALING MOLECULES, CAN

3545
02:38:06,422 --> 02:38:08,691
INTERACT WITH BOTH LIPIDS AND

3546
02:38:08,691 --> 02:38:11,527
PROTEINS, AND ARE OFTEN INVOLVED

3547
02:38:11,527 --> 02:38:17,299
IN REGULATING ACTIVITY AND

3548
02:38:17,299 --> 02:38:18,467
LOCALIZATION.

3549
02:38:18,467 --> 02:38:21,837
WHAT WE KNEW ABOUT THE RFA CRD

3550
02:38:21,837 --> 02:38:23,305
BEFORE WE STARTED THIS PROJECT

3551
02:38:23,305 --> 02:38:27,076
WAS THAT THEY HAVE BEEN SHOWN

3552
02:38:27,076 --> 02:38:37,619
NOT TO BIND ESTER BUT INTERACT 

3553
02:38:40,389 --> 02:38:43,859
WITH STERINE, AND RECENTLY

3554
02:38:43,859 --> 02:38:46,362
INTERACT CORRECTLY WITH RAS.

3555
02:38:46,362 --> 02:38:49,898
THESE FUNCTIONS OF CRD ARE

3556
02:38:49,898 --> 02:38:53,902
CRITICAL UNDER NORMAL SIGNALING

3557
02:38:53,902 --> 02:38:54,203
CONDITIONS.

3558
02:38:54,203 --> 02:39:01,777
BUT IT'S SURPRISING THAT THE CRD

3559
02:39:01,777 --> 02:39:04,646
PLAYS A ROLE IN RAF

3560
02:39:04,646 --> 02:39:07,516
AUTOINHIBITION AS WELL.

3561
02:39:07,516 --> 02:39:09,151
CRD IS IN MAGENTA.

3562
02:39:09,151 --> 02:39:15,090
IT FITS THERE AT THE CENTER OF 

3563
02:39:15,090 --> 02:39:17,259
THE AUTOINHIBITED COMPLEX, MAKES

3564
02:39:17,259 --> 02:39:26,034
CONTEXT WITH KINASE DOMAIN AND 

3565
02:39:26,034 --> 02:39:28,003
BOTH PROTOMERS OF THE DIMER

3566
02:39:28,003 --> 02:39:29,772
WHAT IS THE RELATIVE IMPORTANCE

3567
02:39:29,772 --> 02:39:31,306
AND HOW DID THEY INTEGRATE?

3568
02:39:31,306 --> 02:39:33,041
ONE IS A POSITIVE, ONE IS A

3569
02:39:33,041 --> 02:39:36,211
NEGATIVE, HELPING TO KEEP IT

3570
02:39:36,211 --> 02:39:37,713
EITHER QUIESCENT OR PROMOTING

3571
02:39:37,713 --> 02:39:38,113
ACTIVATION.

3572
02:39:38,113 --> 02:39:44,386
AND WE ALSO WANTED TO KNOW DOO

3573
02:39:44,386 --> 02:39:48,090
RASOPATHY MUTATIONS ALTER THESE

3574
02:39:48,090 --> 02:39:48,490
MUTATIONS.

3575
02:39:48,490 --> 02:39:50,592
SO FOR OUR MUTATIONS WE SELECTED

3576
02:39:50,592 --> 02:39:53,328
A PANEL OF THEM.

3577
02:39:53,328 --> 02:39:54,730
AND ACTUALLY SEVERAL OF THE

3578
02:39:54,730 --> 02:39:57,266
PANEL WERE IN A REGION THAT'S

3579
02:39:57,266 --> 02:39:59,001
KNOWN AS LOOP 1.

3580
02:39:59,001 --> 02:40:01,203
THAT'S ABOUT 11% OF THE

3581
02:40:01,203 --> 02:40:02,271
MUTATIONS CLUSTER IN AND AROUND

3582
02:40:02,271 --> 02:40:03,505
LOOP 1.

3583
02:40:03,505 --> 02:40:06,341
THOSE ARE HIGHLIGHTED IN ORANGE.

3584
02:40:06,341 --> 02:40:09,511
BUT THE ADDITIONAL ONES THAT

3585
02:40:09,511 --> 02:40:10,946
YOU'LL SEE ARE SCATTERED

3586
02:40:10,946 --> 02:40:12,047
THROUGHOUT THE DOMAIN.

3587
02:40:12,047 --> 02:40:14,283
WHAT THEY TEND TO DO IS ADD A

3588
02:40:14,283 --> 02:40:19,121
POSITIVE CHARGE TO THE DOMAIN.

3589
02:40:19,121 --> 02:40:22,357
IN PARTICULAR, THE Q2 57R

3590
02:40:22,357 --> 02:40:27,029
ACCOUNTS FOR 67% OF THE CRD 

3591
02:40:27,029 --> 02:40:29,498
MUTATIONS IN THE RASOPATHY.

3592
02:40:29,498 --> 02:40:35,938
AND SO WHAT WE WANTED TO DO IS

3593
02:40:35,938 --> 02:40:45,180
ASK DO THEY ALTER THE FUNCTION.

3594
02:40:45,180 --> 02:40:47,216
CLEARLY I DON'T KNOW IF THIS

3595
02:40:47,216 --> 02:40:52,821
IS -- SO WE JUST WANTED TO LOOK

3596
02:40:52,821 --> 02:40:55,491
AT THE PANEL WITH THE DIFFERENT

3597
02:40:55,491 --> 02:40:58,760
ACTIVITIES THAT WE KNEW.

3598
02:40:58,760 --> 02:41:00,729
FIRST WAS PS BINDING.

3599
02:41:00,729 --> 02:41:04,766
SHOULD I BE POINTING AT

3600
02:41:04,766 --> 02:41:05,033
SOMETHING?

3601
02:41:05,033 --> 02:41:14,877
IT'S JUST NOT -- YEAH, MAYBE.

3602
02:41:14,877 --> 02:41:19,581
IF I CAN DO THAT MAYBE I HAVE

3603
02:41:19,581 --> 02:41:20,549
ADVANCE THIS DAY.

3604
02:41:20,549 --> 02:41:22,117
TO LOOK AT LIPID BINDING THIS

3605
02:41:22,117 --> 02:41:23,519
WAS A COLLABORATION WITH ANDY

3606
02:41:23,519 --> 02:41:27,222
STEVENS AT THE RAS INITIATIVE,

3607
02:41:27,222 --> 02:41:30,492
WE USED SURFACE PLASMA RESONANCE

3608
02:41:30,492 --> 02:41:31,493
BINDING.

3609
02:41:31,493 --> 02:41:35,330
AND SO WE COUPLED LIPO SENSOR TO

3610
02:41:35,330 --> 02:41:37,933
THE SURFACE, FLOWED OVER,

3611
02:41:37,933 --> 02:41:39,234
PURIFIED CRD, WILDTYPE OR

3612
02:41:39,234 --> 02:41:41,670
MUTANTS, WHAT YOU CAN SEE HERE

3613
02:41:41,670 --> 02:41:49,378
IS THAT IF WE HAD

3614
02:41:49,378 --> 02:41:50,078
PHOSPHO-SERRINE WOULD SEE

3615
02:41:50,078 --> 02:41:55,717
BINDING, AT THE CONTROL

3616
02:41:55,717 --> 02:41:59,321
LIPOSOMES WERE 100% CONTAINED NO

3617
02:41:59,321 --> 02:42:00,856
STEERENE WE SAW VERY LITTLE

3618
02:42:00,856 --> 02:42:01,223
BINDING.

3619
02:42:01,223 --> 02:42:05,594
WE BEGAN TO LOOK AT MUTANTS,

3620
02:42:05,594 --> 02:42:15,437
LOOP 1 VERY ALSO EFFECT ON PS

3621
02:42:15,437 --> 02:42:20,842
BINDING -- VERY LITTLE EFFECT ON

3622
02:42:20,842 --> 02:42:22,077
PS BINDING.

3623
02:42:22,077 --> 02:42:25,714
IF WE STEPPED BACK AND LOOK AT

3624
02:42:25,714 --> 02:42:28,450
STRUCTURE, THIS IS IN

3625
02:42:28,450 --> 02:42:30,018
ELECTROSTATIC SURFACE DEPICTION,

3626
02:42:30,018 --> 02:42:32,721
JUST THE CRD, WE'RE LOOKING AT

3627
02:42:32,721 --> 02:42:36,425
THE MEMBRANE BINDING VIEW,

3628
02:42:36,425 --> 02:42:37,993
THERE'S HYDROPHOBIC POCKET, WHAT

3629
02:42:37,993 --> 02:42:41,897
YOU SEE WITH THESE PUTTING IN

3630
02:42:41,897 --> 02:42:44,633
THESE BASIC RESIDUES, THEN

3631
02:42:44,633 --> 02:42:47,302
SURFACE BECOMES MORE POSITIVELY

3632
02:42:47,302 --> 02:42:47,536
CHARGED.

3633
02:42:47,536 --> 02:42:50,439
AND ALSO SHOWING SIDE VIEW, YOU

3634
02:42:50,439 --> 02:42:52,207
GET THESE BASIC PATCHES BUT THEY

3635
02:42:52,207 --> 02:42:54,242
ARE EXTENDING OUT.

3636
02:42:54,242 --> 02:42:57,012
SO WE THINK THIS ADDING THESE

3637
02:42:57,012 --> 02:42:59,648
POSITIVE CHARGES WOULD BE

3638
02:42:59,648 --> 02:43:02,217
CONDUCIVE FOR INTERACTIONS WITH

3639
02:43:02,217 --> 02:43:04,152
NEGATIVELY CHARGED PLASMA

3640
02:43:04,152 --> 02:43:04,419
MEMBRANE.

3641
02:43:04,419 --> 02:43:05,487
SO THAT MADE SENSE.

3642
02:43:05,487 --> 02:43:11,026
NOW WE WANTED TO LOOK AT EFFECTS

3643
02:43:11,026 --> 02:43:12,527
ON AUTOINHIBITION AND THOUGHT IT

3644
02:43:12,527 --> 02:43:14,229
WAS IMPORTANT TO DO IN THE CELL.

3645
02:43:14,229 --> 02:43:18,800
WE NEEDED TO KNOW HOW IT WAS

3646
02:43:18,800 --> 02:43:19,835
AFFECTING AUTOINHIBITION IN THE

3647
02:43:19,835 --> 02:43:21,003
CONTEXT OF EVERYTHING GOING ON

3648
02:43:21,003 --> 02:43:22,671
IN THE CELL.

3649
02:43:22,671 --> 02:43:26,375
RUSSELL SPENCER SMITH, A

3650
02:43:26,375 --> 02:43:28,243
POSTDOC, DEVELOPED A PROXIMITY

3651
02:43:28,243 --> 02:43:34,316
BASED ASSAY TO DO THIS. 

3652
02:43:34,316 --> 02:43:35,684
IT'S A BIOLUMINESCENCE ENERGY

3653
02:43:35,684 --> 02:43:37,386
TRANSFER BASED ON RESULTS WE HAD

3654
02:43:37,386 --> 02:43:38,820
KNOWN FOR MANY YEARS IN THE

3655
02:43:38,820 --> 02:43:42,024
FIELD, IF YOU JUST EXPRESS THE

3656
02:43:42,024 --> 02:43:45,961
ISOLATE THE BRAF KINASE DOMAIN

3657
02:43:45,961 --> 02:43:47,796
IT CAN FORM CONSTITUATIVE DIMERS

3658
02:43:47,796 --> 02:43:50,232
AND SIGNAL TO MEK.

3659
02:43:50,232 --> 02:43:53,702
IF YOU EXPRESS THE ISOLATED

3660
02:43:53,702 --> 02:43:56,471
REGULATORY DOMAIN, IT COULD BIND

3661
02:43:56,471 --> 02:44:00,342
AND ACT TO BLOCK THE ACTIVITY OF

3662
02:44:00,342 --> 02:44:02,044
THE ISOLATED KINASE DOMAIN.

3663
02:44:02,044 --> 02:44:06,214
SO WE WONDERED IF WE TAGGED EACH

3664
02:44:06,214 --> 02:44:08,216
OF THOSE ISOLATED DOMAINS WITH A

3665
02:44:08,216 --> 02:44:10,819
TAG THAT IN THE RIGHT CONTEXT

3666
02:44:10,819 --> 02:44:13,789
COULD SERVE AS ENERGY DONOR AND

3667
02:44:13,789 --> 02:44:16,692
ENERGY ACCEPTOR, WOULD WE SEE A

3668
02:44:16,692 --> 02:44:20,095
SIGNAL THAT REFLECTED

3669
02:44:20,095 --> 02:44:20,996
AUTOINHIBITION?

3670
02:44:20,996 --> 02:44:24,566
SHOWN HERE AS WE DID INCREASING

3671
02:44:24,566 --> 02:44:25,567
AMOUNTS OF THE REGULATORY

3672
02:44:25,567 --> 02:44:27,202
DOMAIN, HOLDING THE KINASE

3673
02:44:27,202 --> 02:44:29,404
DOMAIN CONSTANT, YOU CAN SEE

3674
02:44:29,404 --> 02:44:34,576
WE'RE SEEING AN INCREASE IN BRET

3675
02:44:34,576 --> 02:44:35,811
SIGNAL THAT CORRELATES WITH

3676
02:44:35,811 --> 02:44:40,382
EXPRESSION OF SIGNALING TO THE

3677
02:44:40,382 --> 02:44:41,616
DOWNSTREAM SUBSTRATE MEK.

3678
02:44:41,616 --> 02:44:44,286
SO THIS WAS GREAT NEWS.

3679
02:44:44,286 --> 02:44:46,621
WE CAN DETECT AUTOINHIBITION.

3680
02:44:46,621 --> 02:44:49,825
WELL, NEXT IMPORTANT THING CAN

3681
02:44:49,825 --> 02:44:52,561
WE DETECT DISRUPTION OF

3682
02:44:52,561 --> 02:44:53,328
AUTOINHIBITION?

3683
02:44:53,328 --> 02:44:54,730
FOR THIS WE STARTED WITH ONE

3684
02:44:54,730 --> 02:44:57,332
THING WE THOUGHT WOULD BE AN

3685
02:44:57,332 --> 02:44:58,133
OBVIOUS CONTROL.

3686
02:44:58,133 --> 02:45:02,337
WHAT IF WE PUT IN AN ACTIVATED

3687
02:45:02,337 --> 02:45:02,871
RAF?

3688
02:45:02,871 --> 02:45:05,640
THAT SHOULD RECRUIT THE RBD AND

3689
02:45:05,640 --> 02:45:11,113
PULL THE REGULATORY DOMAIN AWAY 

3690
02:45:11,113 --> 02:45:11,980
FROM THE KINASE DOMAIN.

3691
02:45:11,980 --> 02:45:14,783
THAT'S WHAT WE SAW.

3692
02:45:14,783 --> 02:45:17,652
ABOUT A 50% REDUCTION IN BRET

3693
02:45:17,652 --> 02:45:18,954
SIGNAL.

3694
02:45:18,954 --> 02:45:21,256
WE DID ANOTHER CONTROL,

3695
02:45:21,256 --> 02:45:25,460
INCORPORATING V600E ONE OF THE

3696
02:45:25,460 --> 02:45:28,296
MOST TRANSFORMING OR ONCOGENIC

3697
02:45:28,296 --> 02:45:29,164
CANCER-ASSOCIATED BRAF

3698
02:45:29,164 --> 02:45:29,531
MUTATIONS.

3699
02:45:29,531 --> 02:45:34,302
AND WHAT WE ALSO FOUND WAS, YES,

3700
02:45:34,302 --> 02:45:35,470
IT ALSO DISRUPTED

3701
02:45:35,470 --> 02:45:36,037
AUTOINHIBITION.

3702
02:45:36,037 --> 02:45:40,208
WHEN WE LOOKED AT MUTANTS IT WAS

3703
02:45:40,208 --> 02:45:42,544
A BIT SURPRISING, ALL THEM

3704
02:45:42,544 --> 02:45:43,812
DISRUPTED ON AUTOINHIBITION

3705
02:45:43,812 --> 02:45:50,185
EXCEPT ONE, THAT WAS THE E275K,

3706
02:45:50,185 --> 02:45:53,021
ABOUT THE TIME WE GOT THE

3707
02:45:53,021 --> 02:45:54,756
STRUCTURE OR FIRST CryoEM

3708
02:45:54,756 --> 02:45:56,491
STRUCTURES, AND WHEN WE STARTED

3709
02:45:56,491 --> 02:45:58,860
LOOKING AT THE STRUCTURES AND

3710
02:45:58,860 --> 02:46:01,163
WHERE THE CRD LAY, IT MADE ALL

3711
02:46:01,163 --> 02:46:04,299
THE SENSE IN THE WORLD BECAUSE

3712
02:46:04,299 --> 02:46:10,138
ALL OF THE POINT MUTATIONS THAT

3713
02:46:10,138 --> 02:46:11,573
DISRUPTED AUTOINHIBITION HAD --

3714
02:46:11,573 --> 02:46:13,675
WERE INTERACTING AND MAKING

3715
02:46:13,675 --> 02:46:18,513
CRITICAL INTERACTIONS THAT 

3716
02:46:18,513 --> 02:46:20,048
HELPED HOLD THE AUTOINHIBITION

3717
02:46:20,048 --> 02:46:22,384
CONFIRMATION TOGETHER.

3718
02:46:22,384 --> 02:46:24,653
WHEN WE LOOKED AT E275 WHERE IT

3719
02:46:24,653 --> 02:46:27,155
LAY, IT'S LIKE OUT THERE WAVING

3720
02:46:27,155 --> 02:46:28,123
IN THE WIND.

3721
02:46:28,123 --> 02:46:31,226
AND SO IT'S NOT SURPRISING THAT

3722
02:46:31,226 --> 02:46:32,394
JUST CHANGING THAT RESIDUE IN

3723
02:46:32,394 --> 02:46:36,765
SIDE CHAINS WOULDN'T HAVE EFFECT

3724
02:46:36,765 --> 02:46:38,333
ON AUTOINHIBITION.

3725
02:46:38,333 --> 02:46:38,667
OKAY.

3726
02:46:38,667 --> 02:46:42,838
SO NOW WHAT ABOUT RAS BINDING?

3727
02:46:42,838 --> 02:46:45,640
SO, WE THEN WENT AND CONTINUED

3728
02:46:45,640 --> 02:46:46,908
TO THINKING IT WAS VERY

3729
02:46:46,908 --> 02:46:48,844
IMPORTANT TO LOOK AT THIS IN THE

3730
02:46:48,844 --> 02:46:50,812
CONTEXT OF THE CELL.

3731
02:46:50,812 --> 02:46:53,248
AND WE HAD ALSO DEVELOPED

3732
02:46:53,248 --> 02:47:00,555
ANOTHER -- THIS IS TRADITIONAL 

3733
02:47:00,555 --> 02:47:04,960
BRET ASSAYS TAGGING VENOUS TO

3734
02:47:04,960 --> 02:47:07,195
KRAS OR THE LOOP, IF THEY ARE

3735
02:47:07,195 --> 02:47:10,699
ABLE TO BIND AN ACTIVATED KRAS

3736
02:47:10,699 --> 02:47:13,435
WE SHOULD SEE A SIGNAL.

3737
02:47:13,435 --> 02:47:16,371
WE ACTUALLY DID A TITRATION

3738
02:47:16,371 --> 02:47:18,740
CURVE AND WE INCORPORATED A

3739
02:47:18,740 --> 02:47:21,943
KNOWN MUTATION IN THE RVD THAT

3740
02:47:21,943 --> 02:47:23,111
COULD DISRUPT BINDING.

3741
02:47:23,111 --> 02:47:25,814
IN CONTEXT OF FULL LENGTH

3742
02:47:25,814 --> 02:47:27,449
PROTEIN, BINDING TO RAS, THIS IS

3743
02:47:27,449 --> 02:47:30,285
GOING TO BE SUBJECT TO EFFECTS

3744
02:47:30,285 --> 02:47:33,521
ON AUTOINHIBITION AS WELL AS TO

3745
02:47:33,521 --> 02:47:36,658
CHANGES IN MEMBRANE BINDING.

3746
02:47:36,658 --> 02:47:38,894
WHAT WE OBSERVED WAS THAT YOU

3747
02:47:38,894 --> 02:47:41,930
SEE LITTLE INCREASE IN RAS

3748
02:47:41,930 --> 02:47:46,368
BINDING IF AUTOINHIBITION IS NOT

3749
02:47:46,368 --> 02:47:46,801
RELIEVED. 

3750
02:47:46,801 --> 02:47:50,939
SO FOR THE MUTANT, E2 75K WHERE

3751
02:47:50,939 --> 02:47:59,214
AUTOINHIBITION WAS  A FINE

3752
02:47:59,214 --> 02:47:59,414
BUNCH.

3753
02:47:59,414 --> 02:48:02,450
THE OTHERS WHERE AUTOINHIBITION

3754
02:48:02,450 --> 02:48:04,386
IS RELEASED BINDING IS HIGHER

3755
02:48:04,386 --> 02:48:06,421
LEVEL, HIGHER AFFINITY AS WELL,

3756
02:48:06,421 --> 02:48:10,158
AT THE TOP OF THE LIST ARE THE 

3757
02:48:10,158 --> 02:48:12,560
ONES THAT HAVE INCREASED

3758
02:48:12,560 --> 02:48:13,528
BINDING.

3759
02:48:13,528 --> 02:48:16,698
IF YOU CAN GET -- CAN OPEN AND

3760
02:48:16,698 --> 02:48:18,333
GET TO RAS, THEN THE ADVANTAGE

3761
02:48:18,333 --> 02:48:20,669
IS GIVEN TO YOU WITH BEING ABLE

3762
02:48:20,669 --> 02:48:28,376
TO HAVE A TIGHTER BINDING.

3763
02:48:28,376 --> 02:48:30,278
THIS IS A SUMMARY OF WHAT

3764
02:48:30,278 --> 02:48:30,912
MUTATIONS DO.

3765
02:48:30,912 --> 02:48:34,215
WE'VE KIND OF BEEN ABLE TO

3766
02:48:34,215 --> 02:48:36,451
CATEGORIZE HOW THEY AFFECTED

3767
02:48:36,451 --> 02:48:38,620
FUNCTIONS OF CRD BUT HOW DOES

3768
02:48:38,620 --> 02:48:41,823
THIS PLAY TOGETHER IN TERMS OF

3769
02:48:41,823 --> 02:48:43,858
THE BIOLOGICAL FUNCTION OF BRAF?

3770
02:48:43,858 --> 02:48:48,530
AND WE FIRST JUST DID A VERY

3771
02:48:48,530 --> 02:48:50,498
STANDARD, YOU KNOW, ASSAY IN

3772
02:48:50,498 --> 02:48:53,601
MAMMALIAN CELLS, LOOKED AT CELL

3773
02:48:53,601 --> 02:48:56,705
GROWTH, FOCUSED ASSAYS, AND

3774
02:48:56,705 --> 02:48:58,440
LOOKED AT SIGNALING DOWNSTREAM,

3775
02:48:58,440 --> 02:49:04,145
ABILITY TO ACTIVATE MEK BY

3776
02:49:04,145 --> 02:49:05,981
MEASURING p-MEK LEVELS, THEY

3777
02:49:05,981 --> 02:49:07,382
HAVE ELEVATED ACTIVITY ABOVE

3778
02:49:07,382 --> 02:49:11,353
WILD TYPE BUT WHAT'S NOTICEABLE

3779
02:49:11,353 --> 02:49:14,589
IS THAT THEY E275K THE GUY WHO

3780
02:49:14,589 --> 02:49:18,893
CAN ONLY -- HAS HIGHER BINDING

3781
02:49:18,893 --> 02:49:20,061
AFFINITY BUT DOESN'T RELIEVE

3782
02:49:20,061 --> 02:49:20,996
AUTOINHIBITION IS THE WEAKEST OF

3783
02:49:20,996 --> 02:49:24,099
ALL OF THE OTHER MUTANTS.

3784
02:49:24,099 --> 02:49:27,002
SO WE THOUGHT IT WAS IMPORTANT

3785
02:49:27,002 --> 02:49:29,871
ACTUALLY TO TRY TO GET INTO A

3786
02:49:29,871 --> 02:49:31,506
SYSTEM WHICH WOULD BE MORE

3787
02:49:31,506 --> 02:49:36,945
RELATABLE TO THE PATIENT. 

3788
02:49:36,945 --> 02:49:38,380
WHAT PATIENTS WITH EXPERIENCING,

3789
02:49:38,380 --> 02:49:45,954
WORKING WITH NCI AND FREDERICK

3790
02:49:45,954 --> 02:49:49,357
AND BIGGER LABORATORY, TO

3791
02:49:49,357 --> 02:49:51,026
ESTABLISH SOME ASSAYS IN

3792
02:49:51,026 --> 02:49:55,063
ZEBRAFISH, MODEL SYSTEM OF

3793
02:49:55,063 --> 02:49:55,764
ZEBRAFISH, THAT WOULD

3794
02:49:55,764 --> 02:49:58,400
RECAPITULATE SOME THINGS SEEN IN

3795
02:49:58,400 --> 02:49:59,901
THE RASOPATHY PATIENTS.

3796
02:49:59,901 --> 02:50:05,740
SO, WE INJECT THE PROTEINS OR

3797
02:50:05,740 --> 02:50:07,942
RNA INTO THE ZEBRAFISH AT THE

3798
02:50:07,942 --> 02:50:11,713
ONE CELL STAIN, ONE DAY

3799
02:50:11,713 --> 02:50:16,484
POST-FERTILIZATION LOOK AT --

3800
02:50:16,484 --> 02:50:18,953
MEASURE EMBRYS, AXES, LOOKING

3801
02:50:18,953 --> 02:50:21,790
FOR DEFECTS IN CONVERGENT

3802
02:50:21,790 --> 02:50:24,826
EXTENSION MOVEMENTS REFLECTIVE

3803
02:50:24,826 --> 02:50:27,595
IN GETTING OBLONG EMBRYOS.

3804
02:50:27,595 --> 02:50:30,498
WE ALSO INJECTED AND LOOKED AT

3805
02:50:30,498 --> 02:50:32,400
THREE DAYS POST-FERTILIZATION,

3806
02:50:32,400 --> 02:50:34,269
WHERE YOU CAN ACTUALLY SEE HEART

3807
02:50:34,269 --> 02:50:36,604
EDEMA THAT MANY OF THE RASOPATHY

3808
02:50:36,604 --> 02:50:38,540
PATIENTS HAVE.

3809
02:50:38,540 --> 02:50:43,611
AND FINALLY WE ALSO LOOK AT

3810
02:50:43,611 --> 02:50:46,815
ANGLE OF THE CARTILAGE WHICH IS

3811
02:50:46,815 --> 02:50:50,318
A DIRECT MEASURE, INDICATIVE OF

3812
02:50:50,318 --> 02:50:52,520
GETTING INCREASED HEAD WIDTH AND

3813
02:50:52,520 --> 02:50:54,989
EYE SPACING, CHARACTERISTICS OF

3814
02:50:54,989 --> 02:50:56,558
THESE PATIENTS.

3815
02:50:56,558 --> 02:50:57,892
AND SO WE TOOK REPRESENTATIVE

3816
02:50:57,892 --> 02:51:00,395
FROM EACH OF OUR KIND OF CLASSES

3817
02:51:00,395 --> 02:51:02,997
OF THESE MUTANTS, AND WHAT YOU

3818
02:51:02,997 --> 02:51:08,336
CAN SEE HERE IS THAT MUTATIONS

3819
02:51:08,336 --> 02:51:09,637
THAT RELIEVE AUTOINHIBITION SHOW

3820
02:51:09,637 --> 02:51:11,239
GREATEST INCREASE IN CONVERGENT

3821
02:51:11,239 --> 02:51:12,207
EXTENSION MOVEMENTS SO THESE

3822
02:51:12,207 --> 02:51:16,945
CELLS ARE MOVING MORE THAN THEY

3823
02:51:16,945 --> 02:51:22,750
REALLY SHOULD DURING

3824
02:51:22,750 --> 02:51:23,585
GASTRILATION.

3825
02:51:23,585 --> 02:51:25,720
IF WE LOOK AT HEART EDEMA

3826
02:51:25,720 --> 02:51:35,964
MUTANTS THAT RELIEVE

3827
02:51:35,964 --> 02:51:36,598
AUTOINHIBITION INCREASE, CLEARLY

3828
02:51:36,598 --> 02:51:41,769
DIFFERENT, NOT AS SEVERE AS WITH

3829
02:51:41,769 --> 02:51:43,805
THE OTHER TWO.

3830
02:51:43,805 --> 02:51:46,040
WHEN WE LOOK AT ANGLE OF THE

3831
02:51:46,040 --> 02:51:50,879
CARTILAGE YOU CAN SEE FOR THE

3832
02:51:50,879 --> 02:51:52,247
E275K IT IS NO DIFFERENT THAN

3833
02:51:52,247 --> 02:51:53,248
WILD TYPE.

3834
02:51:53,248 --> 02:51:56,184
FOR THE OTHER TWO WITH RELIEF

3835
02:51:56,184 --> 02:51:59,287
AND AUTOINHIBITION YOU'RE SEEING

3836
02:51:59,287 --> 02:52:01,389
A WIDENING OF THAT ANGLE.

3837
02:52:01,389 --> 02:52:02,557
AND THIS WAS ACTUALLY EXCITING

3838
02:52:02,557 --> 02:52:04,626
TO US BECAUSE WE WENT AND

3839
02:52:04,626 --> 02:52:04,959
LOOKED.

3840
02:52:04,959 --> 02:52:09,464
THERE'S NOT A LOT OF PATIENT 

3841
02:52:09,464 --> 02:52:11,099
DATA FOR THE RASOPATHY PATIENTS

3842
02:52:11,099 --> 02:52:13,067
BUT THERE'S SOME.

3843
02:52:13,067 --> 02:52:16,337
IN PARTICULAR, FOR THIS, E275K,

3844
02:52:16,337 --> 02:52:18,072
ESPECIALLY WHEN YOU'RE LOOKING

3845
02:52:18,072 --> 02:52:20,608
DOWN HERE WITH THE WIDE-SET

3846
02:52:20,608 --> 02:52:22,911
EYES, IT'S REFLECTIVE OF WHAT WE

3847
02:52:22,911 --> 02:52:29,551
SAW IN THE ZEBRAFISH EMBRYOS.

3848
02:52:29,551 --> 02:52:31,519
IT'S A LESS SEVERE DISEASE, IN

3849
02:52:31,519 --> 02:52:33,288
THIS ONE PATIENT, THERE'S ONE

3850
02:52:33,288 --> 02:52:35,356
OTHER PATIENT THAT HAS BEEN

3851
02:52:35,356 --> 02:52:37,692
REPORTED RECENTLY ALTHOUGH WE

3852
02:52:37,692 --> 02:52:39,260
DON'T HAVE MORE INFORMATION

3853
02:52:39,260 --> 02:52:43,998
ABOUT WHAT THEIR TRAITS ARE.

3854
02:52:43,998 --> 02:52:45,166
IT SEEMS ZEBRAFISH, WELL,

3855
02:52:45,166 --> 02:52:47,569
RESULTS IN THE ZEBRAFISH EMBRYOS

3856
02:52:47,569 --> 02:52:50,738
ARE CORRELATING WELL WITH

3857
02:52:50,738 --> 02:52:52,874
PHENOTYPES OBSERVED IN PATIENTS.

3858
02:52:52,874 --> 02:52:57,645
AND SO, YOU KNOW, WE THINK

3859
02:52:57,645 --> 02:53:04,552
ACTUALLY TAKING THIS I GUESS

3860
02:53:04,552 --> 02:53:06,120
BIOCHEMICAL CELL, BIOLOGICAL,

3861
02:53:06,120 --> 02:53:09,524
STRUCTURAL APPROACH, HAS BEEN

3862
02:53:09,524 --> 02:53:12,594
VERY INFORMATIVE AND STUDYING

3863
02:53:12,594 --> 02:53:14,596
THESE DISEASE-ASSOCIATE MUTANTS

3864
02:53:14,596 --> 02:53:16,264
CAN GIVE IMPORTANT REGULATORY

3865
02:53:16,264 --> 02:53:16,564
INFORMATION.

3866
02:53:16,564 --> 02:53:18,533
AND I THINK IT'S INTERESTING TO

3867
02:53:18,533 --> 02:53:20,535
THINK ABOUT THIS IN CANCER WHEN

3868
02:53:20,535 --> 02:53:24,005
YOU REALLY NEED TO BE HITTING

3869
02:53:24,005 --> 02:53:26,074
THE CELL WITH A SLEDGEHAMMER,

3870
02:53:26,074 --> 02:53:28,409
THOSE MUTATIONS ACTIVATE THE

3871
02:53:28,409 --> 02:53:31,913
CATALYTIC ACTIVITY, YES, THEY

3872
02:53:31,913 --> 02:53:33,081
ALSO RELIEVE AUTOINHIBITION, BUT

3873
02:53:33,081 --> 02:53:34,148
WHILE THEY ARE SUCH STRONG

3874
02:53:34,148 --> 02:53:37,485
DRIVERS IS BECAUSE OF ENHANCED

3875
02:53:37,485 --> 02:53:38,953
CATALYTIC ACTIVITY, WHERE WHEN

3876
02:53:38,953 --> 02:53:41,356
YOU HAVE TO SLIGHTLY UPREGULATE

3877
02:53:41,356 --> 02:53:43,891
THE PATHWAY, IN ORDER TO

3878
02:53:43,891 --> 02:53:47,428
MAINTAIN LIFE, THESE MUTATIONS

3879
02:53:47,428 --> 02:53:48,363
ARE ACTUALLY IMPACTING

3880
02:53:48,363 --> 02:53:50,498
MECHANISMS THAT ARE KNOWN TO

3881
02:53:50,498 --> 02:53:57,105
KEEP THE PROTEIN IN A QUIESCENT 

3882
02:53:57,105 --> 02:53:58,473
STATE WHEN SIGNALING IS NOT

3883
02:53:58,473 --> 02:53:58,873
GOING ON.

3884
02:53:58,873 --> 02:54:01,643
WE'RE EXCITED TO BE PART OF A

3885
02:54:01,643 --> 02:54:06,848
NEW INITIATIVE CALLED THE NCI

3886
02:54:06,848 --> 02:54:07,782
ADVANCING RASOPATHY THERAPY,

3887
02:54:07,782 --> 02:54:11,552
ART, BRINGING TOGETHER PEOPLE

3888
02:54:11,552 --> 02:54:17,358
FROM THE CLINICAL GENETICS, 

3889
02:54:17,358 --> 02:54:19,394
PEDIATRIC ONCOLOGY, NCI, NCI RAS

3890
02:54:19,394 --> 02:54:22,096
INITIATIVE TO IDENTIFY NEW

3891
02:54:22,096 --> 02:54:25,767
RASOPATHY MUTANTS SO LOOKING

3892
02:54:25,767 --> 02:54:28,403
FIRST AT PATIENT DATA AND SAYING

3893
02:54:28,403 --> 02:54:31,472
ARE THERE MUTATIONS IN SOME OF

3894
02:54:31,472 --> 02:54:33,141
THE KNOWN COMPONENTS THAT

3895
02:54:33,141 --> 02:54:34,309
CONTRIBUTE TO THE RASOPATHYS,

3896
02:54:34,309 --> 02:54:38,012
AND ANOTHER GOAL IS TO ANALYZE

3897
02:54:38,012 --> 02:54:40,315
THESE POTENTIAL NEW MUTANTS AND

3898
02:54:40,315 --> 02:54:41,749
DEVELOPMENT OF NEW THERAPIES.

3899
02:54:41,749 --> 02:54:44,118
I WANT TO SAY WE'VE ALREADY --

3900
02:54:44,118 --> 02:54:45,486
THAT'S WHAT WE'RE CURRENTLY

3901
02:54:45,486 --> 02:54:46,254
WORKING ON.

3902
02:54:46,254 --> 02:54:48,723
WE'VE STARTED WITH THIS ACTUALLY

3903
02:54:48,723 --> 02:54:52,093
LOOKING AT THE CRAF, ONE OF THE

3904
02:54:52,093 --> 02:54:54,262
OTHER MEMBERS OF THE RAS FAMILY

3905
02:54:54,262 --> 02:54:54,562
MEMBERS.

3906
02:54:54,562 --> 02:54:57,632
WE'VE ANALYZED THE PATIENT DATA,

3907
02:54:57,632 --> 02:55:00,435
AND WE FOUND ACTUALLY A COUPLE

3908
02:55:00,435 --> 02:55:01,969
POTENTIAL CLUSTERS, NOW IN THE

3909
02:55:01,969 --> 02:55:04,405
PROCESS OF ANALYZING THESE.

3910
02:55:04,405 --> 02:55:08,376
AGAIN, IT'S INTERESTING,

3911
02:55:08,376 --> 02:55:10,378
REINFORCES THE IDEA THAT BECAUSE

3912
02:55:10,378 --> 02:55:12,246
WHERE THESE CLUSTERS LIE ARE IN

3913
02:55:12,246 --> 02:55:16,617
RESIDUES WE KNOW ARE CRITICAL

3914
02:55:16,617 --> 02:55:17,652
FOR DOWNREGULATING PATHWAY UNDER

3915
02:55:17,652 --> 02:55:21,723
NORMAL GROWTH CONDITIONS. 

3916
02:55:21,723 --> 02:55:24,892
WE THINK THE RASOPATHYS, YOU'RE

3917
02:55:24,892 --> 02:55:27,428
SUBVERTING SOMETHING THAT 

3918
02:55:27,428 --> 02:55:28,062
NORMALLY KEEPS IT SUPPRESSED,

3919
02:55:28,062 --> 02:55:31,199
NOT AS, YOU KNOW, A CATALYTIC

3920
02:55:31,199 --> 02:55:32,066
ACTIVATION.

3921
02:55:32,066 --> 02:55:35,069
SO THERE'S REALLY A DIFFERENCE

3922
02:55:35,069 --> 02:55:35,370
THERE.

3923
02:55:35,370 --> 02:55:38,773
A COUPLE YEARS AGO WE CONDUCTED

3924
02:55:38,773 --> 02:55:40,675
A HIGH-THROUGHPUT SCREEN USING

3925
02:55:40,675 --> 02:55:43,144
OUR LIVE CELL ASSAYS, IN

3926
02:55:43,144 --> 02:55:44,579
COLLABORATION WITH MOLECULAR

3927
02:55:44,579 --> 02:55:46,247
TARGETS PROGRAM IN FREDERICK,

3928
02:55:46,247 --> 02:55:48,449
AND THEIR LARGE AND DIVERSE

3929
02:55:48,449 --> 02:55:50,952
LIBRARY OF NATURAL PRODUCTS.

3930
02:55:50,952 --> 02:55:53,321
WE'RE LOOKING FOR COMPOUNDS THAT

3931
02:55:53,321 --> 02:55:54,822
WOULD DISRUPT DIRECTLY OR

3932
02:55:54,822 --> 02:55:56,824
INDIRECTLY SOME OF THESE

3933
02:55:56,824 --> 02:56:03,731
CRITICAL PROTEIN INTERACTIONS 

3934
02:56:03,731 --> 02:56:05,433
LIKE RAF, RAF BINDING, RAF

3935
02:56:05,433 --> 02:56:06,401
DIMERIZATION.

3936
02:56:06,401 --> 02:56:09,570
I WANT TO SHOW HOW WE'VE USED A

3937
02:56:09,570 --> 02:56:11,472
NATURAL PRODUCT INHIBITOR, AGAIN

3938
02:56:11,472 --> 02:56:13,841
SHOWING A MUTANT, AN N-RAS

3939
02:56:13,841 --> 02:56:16,544
MUTANT, REMEMBER RAS IS UPSTREAM

3940
02:56:16,544 --> 02:56:18,713
FROM RAF, WE KNOW SIGNAL

3941
02:56:18,713 --> 02:56:21,482
TRANSMISSION FROM A MUTANT RAF

3942
02:56:21,482 --> 02:56:22,750
DOES REQUIRE RAF DIMERIZATION.

3943
02:56:22,750 --> 02:56:26,354
AND YOU CAN SEE THAT EXPRESSION

3944
02:56:26,354 --> 02:56:29,791
OF THIS MUTANT IN THE ZEBRAFISH

3945
02:56:29,791 --> 02:56:31,592
EMBRYOS, YOU GET ELONGATED

3946
02:56:31,592 --> 02:56:32,293
EMBRYO.

3947
02:56:32,293 --> 02:56:34,762
AND IF WE TREAT THEM WITH THIS,

3948
02:56:34,762 --> 02:56:38,399
AS I CALL IT A RAF DIMER

3949
02:56:38,399 --> 02:56:39,901
DISRUPTER, WE CAN ACTUALLY

3950
02:56:39,901 --> 02:56:41,903
SUPPRESS THAT PHENOTYPE.

3951
02:56:41,903 --> 02:56:43,638
SO WE'RE STARTING TO SEE

3952
02:56:43,638 --> 02:56:43,938
SUPPRESSION.

3953
02:56:43,938 --> 02:56:47,275
IT'S TURNING OUT TO BE A VERY

3954
02:56:47,275 --> 02:56:49,610
RAPID WAY OF ASSESSING IF ANY OF

3955
02:56:49,610 --> 02:56:53,314
THESE NEW DRUGS OR NEW COMPOUNDS

3956
02:56:53,314 --> 02:56:57,018
MAY HAVE AN EFFECT ON SOME

3957
02:56:57,018 --> 02:56:58,953
RASOPATHY MUTANTS THAT ARE  NOT

3958
02:56:58,953 --> 02:56:59,954
-- YOU KNOW, THEIR ACTIVITY

3959
02:56:59,954 --> 02:57:01,355
NEEDS TO BE DAMPENED DOWN.

3960
02:57:01,355 --> 02:57:03,724
IT DOESN'T HAVE TO BE COMPLETELY

3961
02:57:03,724 --> 02:57:05,092
TURNED OFF.

3962
02:57:05,092 --> 02:57:06,494
JUST DAMPENED DOWN, YOU KNOW, TO

3963
02:57:06,494 --> 02:57:11,766
HELP SOME OF THESE PATIENTS.

3964
02:57:11,766 --> 02:57:12,800
I'D LIKE TO THANK -- THERE ARE

3965
02:57:12,800 --> 02:57:16,704
THE PEOPLE IN THE LAB THAT

3966
02:57:16,704 --> 02:57:18,206
HELPED, COLLABORATORS, AND LIKE

3967
02:57:18,206 --> 02:57:20,575
EVERYONE ELSE, I THINK I HAVE

3968
02:57:20,575 --> 02:57:25,446
TO --  A DEBT OF GRATITUDE TO

3969
02:57:25,446 --> 02:57:27,682
THE INSTITUTE BECAUSE WHAT WE'VE

3970
02:57:27,682 --> 02:57:29,984
ACCOMPLISHED OVER THE YEARS IS

3971
02:57:29,984 --> 02:57:32,720
REALLY REFLECTIVE OF A GROUP

3972
02:57:32,720 --> 02:57:34,188
OF -- A TEAM OF COLLABORATORS

3973
02:57:34,188 --> 02:57:36,224
I'VE BEEN ABLE TO WORK WITH WHO

3974
02:57:36,224 --> 02:57:37,558
HAVE ALWAYS BEEN RECEPTIVE, YOU

3975
02:57:37,558 --> 02:57:39,694
KNOW, TO SAYING, SURE, COME TO

3976
02:57:39,694 --> 02:57:42,930
OUR LAB, YOU KNOW, WE'LL BE GLAD

3977
02:57:42,930 --> 02:57:44,465
TO HELP YOU DO THIS.

3978
02:57:44,465 --> 02:57:47,869
I'M REALLY GRATEFUL FOR HAVING

3979
02:57:47,869 --> 02:57:53,140
TO BE HERE, BASICALLY MY ENTIRE

3980
02:57:53,140 --> 02:57:55,343
CAREER AS AN INDEPENDENT

3981
02:57:55,343 --> 02:57:55,676
INVESTIGATOR.

3982
02:57:55,676 --> 02:57:58,379
SO THANK YOU.

3983
02:57:58,379 --> 02:57:58,646
[APPLAUSE]

3984
02:57:58,646 --> 02:58:04,385
 

3985
02:58:04,385 --> 02:58:04,685
 

3986
02:58:04,685 --> 02:58:07,922
>> THAT CONCLUDES THIS

3987
02:58:07,922 --> 02:58:09,123
PHENOMENAL SYMPOSIUM.

3988
02:58:09,123 --> 02:58:12,293
BEFORE I THANK THE SPEAKERS, I'M

3989
02:58:12,293 --> 02:58:14,762
GOING TO MAKE A SUGGESTION THAT

3990
02:58:14,762 --> 02:58:18,533
NEXT YEAR WE CALL THIS SYMPOSIUM

3991
02:58:18,533 --> 02:58:20,835
MYTH BUSTERS BECAUSE I THINK

3992
02:58:20,835 --> 02:58:23,037
WHAT WE'VE LEARNED, SKIN FLORA

3993
02:58:23,037 --> 02:58:28,676
CAN BE FRIEND AND FOE, WE'VE

3994
02:58:28,676 --> 02:58:30,378
LEARNED THAT YOU CAN BREAK DNA

3995
02:58:30,378 --> 02:58:33,080
IN THE COURSE OF MEIOSIS, AND

3996
02:58:33,080 --> 02:58:35,049
THEN USE THE MITOTIC APPARATUS

3997
02:58:35,049 --> 02:58:35,783
TO FIX IT.

3998
02:58:35,783 --> 02:58:38,553
WE'VE LEARNED THAT YOU CAN HAVE

3999
02:58:38,553 --> 02:58:41,355
A POST MITOTIC CELL, AND HAVE

4000
02:58:41,355 --> 02:58:45,760
DAMAGE DONE TO IT BY AN

4001
02:58:45,760 --> 02:58:49,664
ANTI-MITOTIC AGENT.

4002
02:58:49,664 --> 02:58:52,199
WE'VE LEARNED THAT THERE ARE

4003
02:58:52,199 --> 02:58:54,702
PATIENTS WHO HAVE ABERRATIONS OF

4004
02:58:54,702 --> 02:58:59,206
SORT OF GAIN OF FUNCTION

4005
02:58:59,206 --> 02:58:59,774
INFLAMMATORY ABERRATIONS OF

4006
02:58:59,774 --> 02:59:00,408
GAMMA INTERFERON.

4007
02:59:00,408 --> 02:59:02,877
AND YOU CAN GIVE THEM INTERFERON

4008
02:59:02,877 --> 02:59:04,045
TO MAKE THEM BETTER.

4009
02:59:04,045 --> 02:59:06,514
AND WE'VE LEARNED THAT YOU CAN

4010
02:59:06,514 --> 02:59:12,219
HAVE A PEPTIDE THAT INHIBITS THE

4011
02:59:12,219 --> 02:59:14,622
FUNCTION OF A SIGNAL

4012
02:59:14,622 --> 02:59:17,258
TRANSDUCTANT, AND IT CAN ALSO

4013
02:59:17,258 --> 02:59:19,694
ACTIVATE THAT SAME SIGNAL

4014
02:59:19,694 --> 02:59:20,995
TRANSDUCTANT AND SO I THINK

4015
02:59:20,995 --> 02:59:23,631
WE'VE LEARNED THAT THERE'S A LOT

4016
02:59:23,631 --> 02:59:26,100
MORE COMPLEXITY TO THE BIOLOGICS

4017
02:59:26,100 --> 02:59:27,935
SYSTEMS THAN WE EVER EXPECTED

4018
02:59:27,935 --> 02:59:30,805
AND I THANK ALL OF OUR

4019
02:59:30,805 --> 02:59:32,340
WONDERFUL, WONDERFUL SPEAKERS

4020
02:59:32,340 --> 02:59:34,342
FOR HELPING US UNDERSTAND THAT.

4021
02:59:34,342 --> 02:59:36,577
PLEASE JOIN ME IN CONGRATULATING

4022
02:59:36,577 --> 02:59:36,911
OUR SPEAKERS.

4023
02:59:36,911 --> 02:59:46,911
[APPLAUSE]