SO, GOOD MORNING, EVERYONE.
MY NAME IS MICHAEL GOTTESMAN,
CHIEF OF THE LABORATORY OF CELL
BIOLOGY AT THE NCI AND THE
FORMER DEPUTY DIRECTOR FOR
INTRAMURAL RESEARCH AT THE NIH.
AND I'VE KNOWN AND KNEW PHIL FOR
MANY, MANY YEARS.
ON BEHALF OF THE NIH ACTING
DIRECTOR, DR. LARRY TABAK AND
CURRENT DEPUTY DIRECTOR NINA
SCHOR I'D LIKE TO WELCOME YOU TO
THE SYMPOSIUM HONORING THE
EXTRAORDINARY ACCOMPLISHMENTS OF
PHIL LEDER, A SCIENTIST BOTH AT
THE NIH AND AT HARVARD.
AS YOU'LL SEE, WAS INSTRUMENTAL
IN PROBABLY MOST OF THE MAJOR
MOLECULAR ACCOMPLISHMENTS OF THE
LAST 50 YEARS, AND IN GENERATING
AN INCREDIBLE NUMBER OF TRAINEE
TRAINEES.
THERE WILL BE NINE FORM MEMBERS
OF THE LABORATORY, THE TENTH
COULDN'T MAKE IT, WE'RE SORRY
ABOUT THAT.
IT GIVES MORE TIME I THINK AND
ALLOWS US TO PUT A BREAK IN
WHICH WE DIDN'T HAVE SCHEDULED
BEFORE.
I WOULD LIKE TO WELCOME THE
LEDER FAMILY.
LET'S SEE, THERE YOU ARE!
I KNOW IRIS IS NOT HERE BUT WILL
WATCH REMOTELY, AND THEIR
CHILDREN AND PHIL'S CHILDREN,
AND I WELCOME YOU AND THANK YOU
FOR BEING GENEROUS PARTICIPANTS
IN THIS PROGRAM.
I ALSO WANT TO THANK THE FAES
WHICH IS SPONSORING THE PROGRAM,
HOSTING THIS ACTIVITY, WE'RE
VERY GRATEFUL AND YOU'LL HEAR
MORE LATER FROM CHRISTINA ABOUT
THE FAES.
AND THE ORGANIZING COMMITTEE WHO
HELPED COME UP WITH THE PROGRAM.
I'VE NEVER BEEN INVOLVED IN AN
ACTIVITY IN WHICH EVERY SINGLE
PERSON WHO WAS ASKED TO SPEAK
SAID ABSOLUTELY YES, I WILL
COME.
SO, THIS IS I THINK A TESTAMENT
TO PHIL'S LEGACY AS WELL.
I SEE THERE ARE SOME YOUNGER
CAREER SCIENTISTS IN THE
AUDIENCE, AND SO LET ME TELL YOU
ABOUT PHIL LEDER.
MOST OF US KNOW QUITE A LOT
ABOUT HIS ACCOMPLISHMENTSES BUT
IF YOU WERE BORN AFTER 1980 YOU
MIGHT NOT REALIZE HOW MUCH HE
DID.
UNDERGRADUATE AT HARVARD
COLLEGE, M.D. HARVARD MEDICAL
SCHOOL, DURING COLLEGE ABOUT
INTERNSHIP WITH MARTHA VAUGHN,
NATIONAL HEART INSTITUTE, A
LEGENDARY NIH SCIENTIST AND I
THINK HE LEARNED REALLY
WONDERFUL BIOCHEMISTRY FROM HER.
AFTER HIS MEDICAL INTERNSHIP AND
RESIDENCY AT THE UNIVERSITY OF
MINNESOTA, HE RETURNED TO NIH TO
DO POSTDOCTORAL WORK WITH
MARSHAL NUREMBERG AND DESIGNED
EXPERIMENTS THAT MADE POSSIBLE
DECIPHERING OF GENETIC CODE,
WEIZMANN INSTITUTES IN ISRAEL,
CAME BACK TO NIH, MOVED TO NICHD
TO BECOME CHIEF OF LABORATORY OF
MOLECULAR GENETICS, AND THIS
WAS A REALLY LANDMARK
CONTRIBUTION TO THE MOLECULAR
BIOLOGY.
HE ALSO ASKED ANOTHER IMPORTANT
QUESTION, HOW IS IT POSSIBLE
THAT THERE'S SO MUCH ANTIBODY
DIVERSITY IF THE NUMBER OF GENES
ARE SO LIMITED, AND BEGAN
WORKING ON THAT AT NIH AS WELL.
HIS INTEREST IN TEACHING WAS
EXEMPLIFIED BY STINT AS
PRESIDENT FOR ADVANCED EDUCATION
IN SCIENCE, OUR HOST FOR TODAY,
HEAD OF THE FAES CHEMISTRY
DEPARTMENT.
IN 1980 WAS RECRUITED TO HARVARD
FIRST CHAIR OF MEDICAL GENETICS
DEPARTMENT AND AN DEMONSTRATED
ROLE OF C MICK IN BURKITT'S
LYMPHOMA AND PATENTED THE
ONCOMOUSE, AS YOU KNOW
GENETICALLY ENGINEERED MICE FORM
THE BASIS FOR MUCH OF THE
RESEARCH THAT'S BEEN DONE
RECENTLY, OVER THE YEARS
ACTUALLY, TO UNDERSTAND THE
MOLECULAR BASIS OF CANCER.
PHIL'S WORK WAS RECOGNIZED BY A
GREAT MANY AWARDS, INCLUDING
LASKER MEDICAL RESEARCH AWARD,
NATIONAL MEDAL OF SCIENCE,
ELECTED TO MULTIPLE SOCIETIES
INCLUDING NATIONAL ACADEMIES OF
SCIENCE AND MEDICINE, NATIONAL
ACADEMY OF ARTS AND SCIENCES.
AT NIH AND HARVARD MENTORED AND
INSPIRED A GENERATION OF
RESEARCHERS FORTUNATE ENOUGH TO
SPEND TIME IN HIS LAB.
HE DIED ON FEBRUARY 6, 2020, AT
AGE 85.
SO, WHAT WAS THE SECRET SAUCE
THAT ALLOWED PHIL TO BE SO
SUCCESSFUL?
FIRST OF ALL, HE CHOSE THE MOST
CRITICAL QUESTIONS OF HIS TIME,
AND ANSWERED THEM WITH CAREFULLY
DESIGNED AND BRILLIANTLY
EXECUTED EXPERIMENTS.
AND HE WAS ABLE TO COMMUNICATE
HIS FINDINGS WERE BEAUTIFULLY
PREPARED LECTURES AND PAPERS AND
COULD ATTRACT SOME REALLY
WONDERFUL EARLY CAREER
SCIENTISTS TO WORK WITH HIM,
MANY OF WHOM ARE HERE TODAY.
LOOKING AT TODAY'S PROGRAM, I'M
REMINDED OF A FAMOUS QUOTE FROM
JOHN KENNEDY, WHO OFFICIATED AT
THE WHITE HOUSE OVER A MEETING
OF NOBEL LAUREATES, AND MANY OF
YOU MAY REMEMBER THIS QUOTE AND
COMMENTED, I THINK THIS IS THE
MOST EXTRAORDINARY COLLECTION OF
TALENT OF HUMAN KNOWLEDGE THAT
HAS EVER BEEN GATHERED TOGETHER
AT THE WHITE HOUSE, WITH THE
POSSIBLE EXCEPTION OF WHEN
THOMAS JEFFERSON DINED ALONE.
SO OUR SPEAKERS TODAY ARE AN
AMAZING COLLECTION OF TALENT AT
THE NIH, ONLY EXCEEDED BY THE
TIME WHEN PHIL LEDER WAS CHIEF
OF THE LABORATORY OF MOLECULAR
GENETICS AT NICHD.
LET ME FINISH WITH A PERSONAL
ANECDOTE.
I MENTIONED HE WAS A DEDICATED
TEACHER AND TAUGHT A VERY
POPULAR FAES COURSE, DNA
REPLICATION, TRANSCRIPTION, AND
TRANSLATION.
DURING MY POSTDOCTORAL YEARS AT
NIH MY MENTOR, MARTY GELLER,
ASKED ME ONE YEAR TO TAKE OVER
THE TEACHING OF THE DNA
REPLICATION PART OF THE COURSE
BECAUSE HE COULD NOT DO IT.
I WOULD MEET REGULARLY WITH PHIL
TO DISCUSS THE CURRICULUM, WHAT
I WOULD BE PRESENTING, WHAT HE
WOULD SUBSEQUENTLY BE
PRESENTED.
ONE WARM SUMMER DAY AS I WAS
TALKING TO HIM ON THE PHONE I
NOTICED THERE SOUNDED LIKE AN
ECHO, HE COULD HEAR HIS VOICE
TWICE, IN DIFFERENT BUILDINGS A
FEW HUNDRED YARDS APART, WE USED
TO BE ABLE TO OPEN THE WINDOWS,
CAN YOU IMAGINE?
HE WAS A FEW HUNDRED YARDS AWAY.
I REALIZED I COULD HEAR HIM ON
THE PHONE BUT ALSO COULD HEAR
HIS VOICE PROJECTING ACROSS THE
SPACE BETWEEN THE TWO BUILDINGS.
HE HAD A BEAUTIFUL SPEAKING
VOICE, WHICH PROJECTED EXTREMELY
WELL, I'M SURE YOU ALL REMEMBER
THE TALKS THAT HE GAVE.
SO HOW CAN RERECOGNIZE SUCH
IMMENSE TALENT?
THIS SYMPOSIUM IS ONE WAY TO
HIGHLIGHT PHIL LEDER'S LEGACY OF
SCIENCE AND TRAINING AND WITH
THE HELP OF DONORS FAES DONATED
A HOUSE ACROSS ON CEDAR LANE IN
HIS NAME.
I'D LIKE TO INTRODUCE THE CEO
AND EXECUTIVE DIRECTOR OF THE
FAES WHO WILL TELL YOU MORE
ABOUT THIS PROJECT.
CHRISTINA?
[APPLAUSE]
>> GOOD MORNING, AND THANK YOU
SO VERY MUCH FOR BEING HERE WITH
US TODAY.
MY NAME IS CHRISTINA, IN MY 12
YEARS HERE WORKING WITH THE NIH
COMMUNITY, I HAVE NEVER BEEN
MORE PROUD.
FAES WAS CREATED IN 1959 BY NIH
SCIENTISTS FOR THE NIH AND
BROADER BIOMEDICAL COMMUNITY,
OVER THE LAST 64 YEARS WE HAVE
TRAINED AND EDUCATED MORE THAN
150,000 INDIVIDUALS.
SOME PEOPLE THINK OF US AS A
SLEEPING GIANT, A SMALL
ORGANIZATION, BUT WE GET TO HAVE
SUCH AN AMAZING BROAD IMPACT.
AND WHEN I WAS DOING MY RESEARCH
ABOUT DR. PHIL LEDER, I WAS
TREMENDOUSLY PROUD AND HUMBLED
BY NOT ONLY HIS ACCOMPLISHMENTS
AT NIH OF COURSE AND BEYOND, BUT
THE FACT HE SERVED AS ONE OF OUR
BOARD PRESIDENTS.
WE'VE HAD THE PRIVILEGE OF
HAVING WONDERFUL PRESIDENTS AND
BOARD MEMBERS BUT I HAVE TO SAY
PHIL LEDER HAS DONE SOME
SPECTACULAR THINGS, AND I THINK
IT'S CERTAINLY EVIDENT, I'M
GOING TO ECHO WHAT DR. GOTTESMAN
SAID, EVIDENCED BY THE SPEAKERS
YOU'LL HEAR TODAY.
WE WERE SO EXCITED WE THOUGHT WE
WOULD EXTEND ALL INVITATIONS AND
WHEN EVERYONE ACCEPTED, I
THOUGHT, OH MY GOODNESS, THESE
WONDERFUL GENIUS PEOPLE NOW HAVE
ONLY 20 MINUTES TO TALK TO YOU.
I'M SURE THEY ARE GOING TO
IMPOSE WONDERFUL WISDOM AND
SHARE ABOUT THE NIH COMMUNITY,
THE BROADER BIOMEDICAL
COMMUNITY, ALSO HARVARD, AND HOW
WORKING TOGETHER AS SCIENTISTS
THE WORLD IS A BETTER PLACE FOR
ALL OF US.
SO THANK YOU SO MUCH FOR BEING
HERE.
WE ARE ALL SO LUCKY TO HAVE
DONORS WHO HELPED US AND WILL BE
DEDICATED ONE OF OUR SCHOLAR
RESIDENCES TO DR. PHIL LEDER.
WE WILL HAVE A WEBSITE UPDATED,
AND A QR CODE IS ON YOUR PROGRAM
AND WILL BE ON THE SCREEN.
PLEASE FOLLOW ALONG BECAUSE THIS
IS GOING TO BE A LIVING STORY.
WE WANT TO NOT ONLY TALK ABOUT
THE HISTORY OF DR. LEDER, WE
WANT TO THANK HIS FAMILY FOR
PARTICIPATING WITH US, AND ALSO
ALL OF THESE WONDERFUL PEOPLER
HERE TODAY.
SO, WITH THAT, WE WILL START OUR
SYMPOSIUM.
THANK YOU ALL FOR BEING HERE.
IT'S REALLY MY HONOR AND
PRIVILEGE.
THANK YOU.
>> I MENTIONED SHIRLEY TILLMAN
COULD NOT BE HERE TODAY.
AS I WAS SPEAKING TO A NUMBER OF
SPEAKERS, EVERYBODY HAD A
SHIRLEY TILLMAN ANECDOTE.
IN HONOR OF SHIRLEY NOT BEING
HERE IF YOU COULD EACH TELL AN
ANECDOTE, I WOULD APPRECIATE IT.
BECAUSE SHE'S NOT HERE, IT MEANS
WE CAN IN ADDITION TO LIMITING
THE SPEAKERS TO 20 MINUTES, WE
CAN HAVE A 20-MINUTE BREAK AND
DEAL WITH THAT AFTER THE FIFTH
SPEAKER, WHICH IS A NATURAL
DIVISION BETWEEN SPEAKERS WHO
WERE WITH PHIL AT HIS NIH LAB
AND THOSE WITH PHIL AT HIS
HARVARD LAB.
NOW, BECAUSE WE DON'T -- SHIRLEY
WAS GOING LEAD A SESSION ON
MENTORING, AND BECAUSE WE WILL
NOT BE HAVING THAT SESSION, IF I
COULD ASK EACH OF THE SPEAKERS,
I THINK THEY ARE AWARE, TO TALK
A LITTLE BIT ABOUT HOW THEIR
EXPERIENCE IN PHIL'S LAB
AFFECTED THEIR SCIENTIFIC
JOURNEYS AND HOPEFULLY WE'LL
HEAR MORE ABOUT THEIR SCIENCE AS
WELL.
AND IF TIME PERMITS AT THE END
WE'LL HAVE A Q&A SESSION.
SO IF YOU HAVE QUESTIONS, HOLD
THEM UNTIL THE END.
NOW, LET'S GET ON WITH THE
PROGRAM.
OUR FIRST SPEAKER IS STUART
ORKIN, DAVID NATHAN
DISTINGUISHED PROFESSOR AT
PEDIATRICS AT HARVARD MEDICAL
SCHOOL.
S TU DEFINED MOLECULAR BASIS OF
BLOOD DISORDERS, ESPECIALLY
THALASSEMIAS, B.S. FROM M.I.T.,
M.D. HARVARD, RESEARCH ASSOCIATE
WITH PHIL LEDER AT NIH FROM 1973
TO 1975.
FOLLOWING HIS EXPERIENCE AT NIH,
STU RETURNED TO BOSTON, TRAINED
IN PEDIATRICS AND HEMATOLOGY
ONCOLOGY, CHAIRMAN OF DEPARTMENT
OF PEDIATRIC ONCOLOGY AT
DANA-FARBER CANCER INSTITUTE,
ELECTED MEMBER, THIS WILL SOUND
FAMILIAR, NATIONAL ACADEMIES OF
SCIENCE AND MEDICINE, AMERICAN
ACADEMY OF ARTS AND SCIENCES,
AMERICAN PHILOSOPHICAL SOCIETY,
WON MANY AWARDS INCLUDING
ALPERT, GRUBER PRIZE, CANADIAN
INTERNATIONAL AWARD.
STU.
>> THANKS VERY MUCH.
IT'S A PLEASURE AND AN HONOR TO
BE HERE TO CELEBRATE PHIL.
I THINK YOU'LL SEE THAT THERE'S
A DIRECT LINE BETWEEN MY
EXPERIENCE AT THE NIH AND WHAT
I'VE BEEN DOING EVER SINCE.
SO, THIS WAS PHIL PRIOR TO MY
ARRIVAL, I ARRIVED IN 1973.
I GUESS HE HAD A UNIFORM, I
DIDN'T HAVE A UNIFORM.
I WAS AT A PUBLIC HEALTH
SERVICE, BUT I NEVER GOT MY
UNIFORM.
AND THIS IS WHEN I WAS HERE AS
PUBLIC HEALTH SERVICE ASSOCIATE,
BUILDING 6, WHICH WAS A QUARTER
OF THE SIZE IT IS NOW.
THERE'S MY PICTURE FROM THE LAB
NOTEBOOK, I HAD MORE HAIR IN
THOSE DAYS, A PICTURE OF THE
LAB.
IF YOU NOTICE, PHIL IS NOT IN
THIS PICTURE, NOR IS AYA,
BECAUSE THEY ARE ON SABBATICAL
FOR ONE OF THE TWO YEARS I WAS
AT THE NIH.
THAT WAS BEFORE E-MAIL AND ALL
THIS SO IT WAS HARD TO
COMMUNICATE BUT HE CAME BACK AND
FORTH, WE STILL MANAGED TO
COMMUNICATE.
IN 2004, THERE WAS A TRAINING
REUNION AT -- UP IN BOSTON, AND
LOTS OF PEOPLE WERE THERE.
THERE'S SHIRLEY.
SHE TOOK MY LAB BENCH WHEN I
LEFT.
A PICTURE OF ME AND PHIL, TO THE
RIGHT IS PHIL WITH PAUL, A
VISITING SCIENTIST FROM HUNGARY,
PAUL WAS A BIOCHEMISTRY
INSTRUCTOR FOR THE PERSON WHO
JUST WON THE NOBEL PRIZE.
I HAD A CHAIR AT HARVARD NAMED
AFTER ME, DOESN'T HAVE MY NAME
UNTIL I THINK I'M DEAD, PHIL WAS
AT THIS RECEPTION WITH AYA, WE
HAVE A LONG HISTORY.
WHAT I LEARNED FROM PHIL?
TO CHOOSE A GOOD PROBLEM, BE
FOCUSED AND GOAL ORIENTED.
DON'T SWEAT THE SMALL STUFF,
TELL YOUR STORY WELL, AND BE
HUMAN AND AUTHENTIC.
I DON'T HAVE TIME TO TELL
STORIES ABOUT THE LAST POINT BUT
I WAS CLOSE TO FILL BECAUSE OF
SOME PERSONAL THINGS THAT
HAPPENED IN MY FAMILY WHILE I
WAS AT THE NIH, A FEW WEEKS
AFTER I ACTUALLY ARRIVED.
WHAT HAVE I DONE SINCE?
I STUDIED BLOOD CELLS EARLY,
PART OF THE GLOBIN PART OF THE
LAB, WHERE DO THEY COULD FROM,
WHAT DO THEY DO, AND SINCE
LEAVING PHIL, FIRST THING I DID
IN BOSTON WAS DETERMINE
MUTATIONS RESPONSIBLE FOR BETA
THALASSEMIA, AT THE TIME WE
THOUGHT THAT WOULD TELL US ABOUT
RED CELL DEVELOPMENT WHICH IT
RAIL DIDN'T.
CLONED THE FIRST TRANSCRIPTION
FACTOR FOR HEMATOPOIESIS, AND
FOR A LONG PERIOD OF TIME
DEFINED MANY REQUIREMENTS FOR
TRANSCRIPTION FACTORS IN
HEMATOPOIESIS.
WE'VE BEEN WORKING ON THIS
LONGSTANDING PROBLEM IN THE
FIELD, SWITCH FROM FETAL TO
ADULT HEMOGLOBIN, AND I'LL JUST
SUMMARIZE THAT'S BEEN AN ACTIVE
AREA OF LATE.
FOR MANY YEARS BEFORE I EVEN
CAME TO THE NIH, WE KNEW THE
GLOBIN GENES CHANGED, WE HAVE
FETAL GLOBIN, DURING ADULT LIFE
OBVIOUSLY ADULT GLOBIN.
IT WAS RECOGNIZED ACTUALLY IN
1948 BY AN ASTUTE PEDIATRICS
NEWBORNS WITH SICKLE CELL
DISEASE WHEN THEY WERE OLDER
WERE NOT SICK IN UTERO, NOT SICK
IN EARLY NEONATAL PERIOD, AND
SHE SURMISED THIS IS BECAUSE
THEY HAD FETAL HEMOGLOBIN WHICH
WAS PROTECTIVE AND SHE WAS
CORRECT.
THAT LED TO THE NOTION IN THE
FIELD FOR MANY YEARS THAT WE
SHOULD STRIVE TO INCREASE LEVEL
OF FETAL HEMOGLOBIN TO PARTLY
OFFSET THE ADULT HEMOGLOBIN,
BECAUSE GENES ARE EXPRESSED ON
CHROMOSOME THIS WOULD LEAD TO
DECREASE IN ADULT HEMOGLOBIN FOR
THE BETA CHAIN, AND THIS COULD
BE THERAPEUTIC.
AND THE VISION IN THE FIELD FOR
AS LONG AS I'VE BEEN IN IT IS TO
RAISE LEVEL TO ROUGHLY 20% OR SO
IN A CELL ON AVERAGE, REGULATION
IS ACCOMPLISHED, LOCUS CONTROL
REGION IN THE CHROMOSOME 11 IS
THE BASIC DRIVER IN FETAL STAGE
LOOPS TO TWO GAMMA GENES, WHICH
ARE FETAL GENES, IN ADULT BONE
MARROW LOOPS TO THE BETA GENE,
EARLY 2000s WE WERE KIND OF
WANDERING IN THE DESERT BECAUSE
WE HAD A LOT OF INFORMATION
ABOUT HOW GLOBIN GENES WERE
REGULATED, NOTHING ABOUT THE
TEMPORAL OR STAGE-SPECIFIC
EXPRESSION.
AND THE BREAK CAME FROM
ASSOCIATION STUDIES, INVOLVED
WITH A GROUP IN SARDINIA, AND
THIS LED TO THE DISCOVERY OF B
CELL SWITCH FACTOR.
WE WORKED ON THE MECHANISM BY
WHICH IT CONTROLS THE SWITCH,
LEVERAGED FOR GENETIC THERAPIES,
AND I THINK WE DISCOVERED IT BUT
IS IT REALLY IMPORTANT?
THIS WAS A CRITICAL EXPERIMENT,
WITH ENGINEERED MICE, SICKLE
CELL NICE INTERBRED, RESTRICTED
THE KNOCKOUT, ASKED WOULD THAT
RESCUE THE PHENOTYPE.
THE RESULT WAS BETTER THAN WE
MIGHT HAVE GUESSED OR HOPED FOR.
THE SICKLE CELL MOUSE IS IN THE
MIDDLE, MICE WERE RESCUED IN
TERMS OF HEMATOPOIESIS BECAUSE
THEY HAVE FETAL HEMOGLOBIN
EXPRESSION, HIGH LEVEL IN
VIRTUALLY ALL CELLS, REALLY THE
BASIS FOR TREMENDOUS AMOUNT OF
ONGOING CLINICAL WORK RIGHT NOW
IN TRANSLATION CLINICAL TRIALS
FOR BOTH BETA THALASSEMIA AND
SICKLE CELL DISEASE.
IT COMES OUT IN ADULT STAGE AND
DISPLACES ACTIVATORS AND
REPRESSED GAMMA SCREEN, TAKING
PLACE ON A 30 BASE PAIR TEMPLATE
IN THE GAMMAGLOBIN PROMOTER.
BUT THE QUESTION WAS WHAT DOES
GWAS HAVE TO DO?
THERE'S NO CHANGE IN PROTEIN
SEQUENCING COMMON VARIATION BUT
THE VARIATION AFFECTS REGULATORY
ELEMENT ENHANCER WITHIN THE GENE
AND THIS HAPPENS TO BE SPECIFIC
ENHANCER, ADULT SPECIFIC SHOWN
AT THE BOTTOM, ABOUT 13
KILOBASES IN LENGTH, RED CELL
SPECIFIC ELEMENT IN A
STAGE-SPECIFIC AND
ERYTHROID-SPECIFIC MANNER WHICH
GOES ALONG WITH LOTS OF WORK IN
THE GWAS AREA THAT MOST COMMON
VARIATION OCCURS IN REGULATORY
ELEMENTS.
AND FORTUNATELY, ABOUT THIS TIME
CRISPR CAME ON THE SCENE,
ALLOWED US TO DISSECT ENHANCER,
DONE BY PEPPERING ENHANCER AND
ASKING WAS THERE A PLACE IN
13,000 BASE PAIRS WE MIGHT USE
FOR TRANSLATION TO THE CLINIC.
AND REALLY QUITE SURPRISINGLY
THERE'S A SINGLE CITE, ACHILLES
HEEL, WITHIN THE ENHANCER, GATA
BINDING SITE REQUIRED FOR THE
ACTIVITY OF THE ENHANCER TO THE
EXTENT OF ABOUT 2/3 TO 3/4 OF
THE ENHANCER FUNCTION, AND WE
SAID AT THAT TIME THIS WOULD BE
A REALLY IDEAL SITE FOR GENE
EDITING.
SO THIS TURNED OUT TO BE UNIQUE
IN THAT BOTH COMMON VARIATION
AND I HAVEN'T TOLD YOU BUT RARE
GENETIC VARIATION, SO-CALLED
HEREDITARY PERSISTENCE OF FETAL
HEMOGLOBIN SYSTEM CONVERGE,
BINDING SITE AND HOW IT'S
REGULATED.
HERE IS THE ACHILLES HEEL,
SINGLE SITE WITHIN ENHANCER
RESPONSIBLE FOR A GOOD BIT OF
THE ACTIVITY, AND BASICALLY
WHAT'S BEEN TRANSLATED NOW TO
PATIENTS IS SHOWN HERE, THIS IS
TO TAKE HEMATOPOIETIC STEM AND
PROGENITOR CELLS, EDIT THEM AT
THE ACHILLES HEEL, IN THIS CASE
WITH CRISPR/CAS9, AND THEN
REINFUSE THOSE CELLS INTO
PRECONDITIONED PATIENTS, AND
WHAT THIS DOES IS BASICALLY TURN
ON FETAL HEMOGLOBIN IN AN
OTHERWISE ADULT CELL.
WE'RE NOT TURNING THE CELL BACK
TO A FETAL STAGE.
AND THE WAY THIS IS DONE IS
ESSENTIALLY BONE MARROW
TRANSPLANTATION, AND THERE ARE
MANY PARAMETERS WHICH ARE
CRITICAL.
I DON'T HAVE TIME TO GO OVER
REALLY ANY OF THEM EXCEPT TO SAY
THAT THEY NEED TO BE EXECUTED
PRECISELY TO ACCOMPLISH THIS
WELL, AND THIS HAS BEEN
TRANSLATED BY COLLABORATION OF
CRISPR THERAPEUTICS AND BRITAX
PHARMACEUTICAL LEVERAGING
ACHILLES HEEL WITH A SINGLE
GUIDE, THESE ARE THE FIRST TWO
PATIENTS, REPORTED WITH BETA
THALASSEMIA, SICKLE CELL DISEASE
IN THE NEW ENGLAND JOURNAL, BOTH
PATIENTS WERE ESSENTIALLY CURED.
THIS IS FROM AN FDA ADVISORY
MEETING EARLIER THIS WEEK,
BASICALLY YOU CAN SEE AS THEY
CONTINUED THE TRIAL IN SICKLE
CELL PATIENTS THEY ARE FREE OF
PAINFUL CRISES OF SICKLE CELL
DISEASE, MORE THAN 95% OF
PATIENTS HAVE BEEN RENDERED
ESSENTIALLY CURED, AND "NEW YORK
TIMES" COVERED THIS, THIS WEEK,
A FORMAL APPROVAL IS EXPECTED IN
DECEMBER 8, NOT DECEMBER 3,
SHOULD BE APPROVED, THE FIRST
GENETIC EDITING THERAPY.
AND I THINK WE HAVE TO
APPRECIATE HOW PROFOUND THIS IS
FOR THE PATIENTS.
THIS IS ACTUALLY THE FIRST
PATIENT TREATED WHO HAS BEEN
SORT OF A SPOKESPERSON,
BASICALLY SAID HER LIFE -- SHE
DIDN'T HAVE A LIFE BEFORE THIS
THERAPY AND NOW SHE HAS A LIFE
AND ACTUALLY CAN GO ON AND LOOK
TO THE FUTURE.
SO, GENETIC THERAPY IS NOW
VALIDATED.
THE REAL CHALLENGE NOW IS HOW TO
EXPAND THIS TO THE MANY NUMBER
OF PATIENTS WHO COULD BENEFIT
FROM IT, AND QUESTION AS I THINK
COULD BE DONE IN VIVO GENETIC
THERAPY AS OPPOSED TO EX VIVO,
WHICH IS THE CURRENT SCHEME.
THIS HAS BEEN A LONG TIME IN
COMING.
FROM JANET WATSON IN 1948 TO THE
ANTICIPATED APPROVAL AT THE END
OF THIS YEAR, AND IT WAS A LONG
GAP IN BETWEEN.
I THINK I SEE A THREAD RIGHT
BACK TO MY WORK AT THE NIH WITH
PHIL.
I JUST WANT TO IN CLOSING SAY A
FEW THINGS ABOUT WHY THIS IS
REALLY A SOLUTION TO THE
HEMOGLOBIN OPATHIES, AND
ACTIVATION OF HEMOGLOBIN F IS
VALIDATED BY GENETIC, AND
CLINICAL TRIALS, MAINTAINS THE
BALANCE OF GENE EXPRESSION, NOT
THE CASE IN LENTIVIRALGLOBIN
ADDITION PAIN THERAPY, SUITABLE
FOR HEMOGLOBIN OPATHIES, THERE'S
A FIRST GENERATION METHOD BUT I
THINK THERE WILL BE OTHERS
COMING ALONG, MODIFYING THE
BINDING SITE IN THE GAMMA
PROMOTER, USING BASE EDITING
RATHER THAN CRISPR/CAS9, BUT
THESE WILL BE SECOND AND THIRD
GENERATION THERAPIES.
I THINK IT AVOIDS DIFFICULTIES
THAT ARE TECHNICAL IN SINGLE
BASE REPAIR, FOR EXAMPLE, IN
SICKLE CELL DISEASE.
WHAT CONTRIBUTES TO SUCCESS?
MANY UNIQUE ASPECTS OF PROTEIN
AND ITS ROLE WHICH I DON'T HAVE
TIME TO GO INTO, BUT IT'S
REALLY -- THE SYSTEM IS UNIQUELY
HARD WIRED, VERY FEW TARGETS OF
ERYTHROID CELLS OTHER THAN
GLOBINS, CELL IS INSENSITIVE TO
DOWNREGULATION OF THE FACTOR,
NOT TRUE OF MANY TRANSCRIPTION
FACTORS.
THERE'S EXCEEDINGLY ROBUST HUMAN
GENETICS SUPPORTING THE WORK,
STRONG DEPENDS OF HEMOGLOBIN F
ON THIS ONE REPRESSOR, THERE'S A
SECOND REPRESSOR AS WELL THOUGH.
WE HAVE MANY EXPERIMENTAL
RESOURCES IN PLACE, REALLY TO
STUDY THIS FACTOR.
WE HAD MANY OBVIOUSLY
OUTSTANDING COMMITTED TRAINEES,
I ALSO ATTRIBUTE TO DUMB LUCK,
BEING IN THE RIGHT PLACE AT THE
RIGHT TIME.
THIS INVOLVES EX VIVO
MANIPULATION OF CELLS,
PRECONDITIONING, BONE MARROW
TRANSPLANTATION, IT'S GOING TO
BE AT LEAST $2 MILLION, THE
PRICE IS NOT SET BUT I THINK
IT'S A BARGAIN IN TERMS OF
SAVING THESE PATIENTS AND GIVING
THEM A LIFE AND ALSO REDUCING
MEDICAL COSTS.
BUT IT IS CUMBERSOME,
INSUFFICIENT NUMBER OF
TRANSPLANT CENTERS TO TREAT ALL
THE PATIENTS, NOT TRANSFERABLE
TO MANY PARTS OF THE WORLD WHERE
THERE ARE MANY PATIENTS, AND SO
UNLESS WE CAN DO IN VIVO EDITING
AT HIGH EFFICIENCY, I THINK WE
STILL HAVE A NEED FOR SMALL
MOLECULES OR DRUGS TO ACTUALLY
DO WHAT GENETICS CAN DO FOR US.
IT'S BEEN A LONG AND REWARDING
JOURNEY WHERE WE'VE GONE FROM
OVER THE LAST 15 YEARS FROM A
GWAS RIGHT THROUGH TO WHAT WE
BELIEVE WILL BE AN APPROVED
THERAPY BUT I THINK THIS WOULD
NOT HAVE BEEN POSSIBLE WITHOUT,
YOU KNOW, THE MUCH EARLIER WORK,
PARTICULARLY PHIL'S WORK, SO
WE'RE IN A NEW ERA, NEW
TECHNOLOGY IS BEING BROUGHT,
SOME INVOLVING STEM CELLS, AND I
THINK I MUST THANK PHIL AND SAY
HE PROVIDED A GREAT FOUNDATION
FOR GOING FORWARD.
THANK YOU.
[APPLAUSE]
>> THANKS, STU.
WHAT A WONDERFUL EXAMPLE OF HOF
BASIC SCIENCE LEADS TO CLINICAL
IMPROVE.
NEXT SPEAKER IS JONATHAN
SEIDMAN, PROFESSOR OF
CARDIOVASCULAR GENETICS AT
HARVARD, A GRADUATE OF HARVARD
COLLEGE, Ph.D. IN MOLECULAR
BIOLOGY FROM UNIVERSITY OF
WISCONSIN, WAS A POSTDOC IN
PHIL'S LAB AT NIH FROM 1979 TO
1981, WORK REVOLUTIONED
UNDERSTANDING OF GENETIC
DISORDERS THAT LEAD TO CARDIO
CARDIOMYOPATHIES, HE AND HIS
WIFE CRICKET, WHO IS ALSO VERY
WELL KNOWN AT NIH, HAS BEEN A
PARTNER IN MUCH OF THEIR JOINT
RESEARCH, RECOGNIZED BY A NUMBER
OF AWARDS, INCLUDING THE
DISTINGUISHED SCIENTIST AWARD,
AMERICAN HEART ASSOCIATION, KATZ
PRIZE, DISTINGUISHED ACHIEVEMENT
AWARDS FROM SOCIETIES FOR
CARDIOVASCULAR PATHOLOGY,
BRISTOL MYERS SQUIBB AND
McGILL HEART INSTITUTE.
>> THANK YOU, MIKE AM.
GOOD MORNING.
FIRST, THAT'S A TOUGH ACT TO
FOLLOW.
WHAT I'D LIKE TO DO TODAY IS
FIRST OF ALL MENTION THAT ALL
THE WORK I'M GOING TO TALK ABOUT
TODAY WAS DONE BY MYSELF AND
KRICKET SEIDMAN.
YOU'RE TRAINING MANY PEOPLE,
MANY PEOPLE WILL STAY, THE LEDER
HOUSE WILL HAVE PhDs, SOME
MAY HAVE MD/PHD, WE WORKED THIS
OUT IN A BETTER WAY, WAY BETTER
TO GET YOUR M.D. BY MARRIAGE, I
DON'T KNOW IF THAT'S A METHOD
THAT CAN BE BROAD TO THE FAES
BUT I STRONGLY RECOMMEND IT.
I WOULD LIKE TO POINT OUT I WAS
A CO-FOUNDER WITH KRICKET AND
OTHERS OF A SMALL COMPANY
MYOCARDIA, NO LONGER IN
CONFLICT, WHAT I WANT TO DO
ASSUMING I CAN ADVANCE -- YEAH.
DID THAT WORK?
SORRY.
WHAT I WANT TO DO IS TELL YOU
ABOUT THE WORK THAT WE'VE BEEN
DOING, SOMEHOW I WENT TOO FAR.
YEAH, OKAY.
SORRY.
A LITTLE BIT ABOUT LIFE IN
BUILDING 6 IN PHIL LEDER'S LAB,
WHEN I WENT TO PHIL'S LAB IT WAS
AN EYE-OPENING EXPERIENCE FOR
ONE THING PHIL AND I WERE
WORKING TOGETHER, AND THAT WAS
SOMETHING THAT I SAID THAT'S A
GREAT IDEA, HOW COME I DIDN'T
THINK OF THAT?
SOMETHING KRICKET AND I HAVE
TAKEN TO HEART AND HAVE FOLLOWED
EVER SINCE.
BUT IN ADDITION TO THE FACT PHIL
AND I WERE WORKING TOGETHER
THERE WERE -- HIS LAB AND STU
WENT THROUGH SOME OF THE
FEATURES THAT MADE IT A SPECIAL
PLACE, IN PARTICULAR THERE WAS A
SOLUTION BOOK, I DON'T KNOW IF
YOU CAN SEE MY CURSOR, YOU DON'T
FIND IT IN EVERY LABORATORY IN
THE UNITED STATES.
ONLY MOSTLY LABS THAT DESCENDED
FROM PHIL'S LAB HAVE SOLUTION
BOOKS.
AT SOME INSTITUTIONS, THEY ARE
MAKING US CONVERT TO ELECTRONIC
LAB NOTEBOOKS, TRUST ME A
SOLUTION BOOK IS WAY BETTER.
THERE WAS, AS STU POINTED OUT,
THERE'S A PICTURE OF STU AS
EVERYBODY HAD THE SOLUTION
NUMBER, LEN IS GOING TO TELL US
ABOUT HIS, SHIRLEY, I'M SORRY
YOU'RE NOT HERE BECAUSE I WANT
TO POINT OUT NOT ONLY ARE YOU IN
THE SOLUTION BOOK BUT YOU'RE
DOCUMENTED AS BREAKING AN NIH
RULE.
FORTUNATELY FOR ALL OF US THE
STATUTE OF LIMITATIONS HAS
EXPIRED AND THE FACT THAT YOU
WERE CONSUMING ALCOHOL IN
BUILDING 6 IN CELEBRATION, WHICH
HAPPENED MORE FREQUENTLY THAN
YOU MIGHT HAVE EXPECTED, IS JUST
SOMETHING YOU JUST HAVE TO KEEP
TRACK OF.
I WANT TO MENTION WE WERE USING
IN THOSE DAYS A MAJOR
TECHNOLOGY, I WOULD GUESS THAT
HALF THE PEOPLE IN THE AUDIENCE
DON'T KNOW WHAT AN R-LOOP IS.
HERE THEY ARE.
A VERY USEFUL METHOD OF
COMPARING RNA AND DNA SEQUENCES,
NOT DEVELOPED IN PHIL'S LAB, HE
USED IT, HIS LAB USED IT
PRODUCTIVELY FOR SEVERAL YEARS.
STU POINTED OUT A NUMBER OF
RULES.
I THINK THEY ARE PRINCIPLES THAT
WE SHOULD LIVE BY.
PHIL TAUGHT ME A LOT.
HE HAD A VERY STRONG IMPACT ON
THE WAY WE DO SCIENCE.
AND YOU'LL NOTICE HERE THERE'S
NOT A PRINCIPLE NUMBER 1.
ONE OTHER MENTOR POINTED OUT YOU
THEY NEVER HAVE PRINCIPLE NUMBER
1 BECAUSE EVERYBODY HAS ONE.
WE WENT RIGHT TO PRINCIPLE
NUMBER 2.
ONE WAS PEOPLE DON'T CHANGE.
IMPORTANT WHEN YOU LOOK FOR
REVIEWERS FOR YOUR PAPER.
SO YOU WANT PEOPLE WHO ARE GEE
WHIZ PEOPLE, WHO YOU HAVE SHOW
THEM ALMOST ANY RESULT, LIKE THE
NIH SHUTTLE THAT BROUGHT US HERE
TODAY, SOME SAY GEE WHIZ, OTHERS
SAY SO WHAT, BUT YOU WANT A GEE
WHIZ PERSON.
YOU CAN'T DO SCIENCE BY
COMMITTEE.
THAT'S CHANGED OVER THE YEARS,
NOW WE TRIED TO MODIFY THAT WITH
ZOOM, WE NOW HAVE ZOOMS BUT
DOING SCIENCE BY COMMITTEE
DEFINITELY IS A PROBLEM.
I'VE TOLD YOU ABOUT SOLUTION
BOOKS.
ANOTHER RULE THAT I THINK WE
THINK ABOUT QUITE FREQUENTLY,
IT'S NOT WHAT YOU DID FOR ME
YESTERDAY, IT'S NOT THAT PHIL
FOUND CODONS THAT WERE PART OF
THE GENETIC CODE OR THAT HE
IDENTIFIED A METHOD OF CELLULOSE
FOR CAPTURING RNA, TECHNIQUES
STILL USED, VERY IMPORTANT IN
BIOLOGY STILL TODAY, IT'S WHAT
YOU DO FOR ME TODAY AND WHAT
YOU'RE GOING TO DO FOR ME
TOMORROW.
I'M GOING TO DISCUSS, CARRY ON
HOW THOSE RULES PHIL INTRODUCED
ME TO, THE FACT THAT THE SCIENCE
THAT I HAVE DONE FOR THE NEXT 30
YEARS.
SO, AS STU POINTED OUT IN 1980
1980, PHIL MOVED TO BOSTON, THEY
STARTED A NEW GENETICS
DEPARTMENT.
HARVARD MEDICAL SCHOOL NEVER HAD
A GENETICS DEPARTMENT BEFORE
1980, IT'S CHANGED HARVARD
MEDICAL SCHOOL AS WELL AS
CREATING A GREAT PLACE TO DO
SCIENCE.
JUST TO SHOW HOW EFFICIENTLY OR
EFFECTIVELY PHIL'S
ADMINISTRATIVE CAPACITIES WERE
THESE ARE PICTURES OF DEPARTMENT
GROWING FROM 1981 THROUGH 1994.
BY 1994, THE DEPARTMENT WAS SO
BIG THAT WE REALLY COULDN'T FIT
IT INTO A LARGE SINGLE PICTURE
SO WE STOPPED TAKING THESE
PICTURES.
AND PHIL HAD A SERIES OF MORE
PRINCIPLES, FOR GUIDING HIS
DEPARTMENT.
ONE WAS TO RECOGNIZE THE AWESOME
POW EVERY GENETICS, AND THERE
WAS ADMINISTRATIVE THINGS LIKE
DEFINITELY AS A PERSON RUNNING A
LAB YOU HAVE TO LEARN HOW TO SAY
NO IN AN ACADEMIC ENVIRONMENT.
DEVELOPED A SYSTEM, TWO THINGS
MOST IMPORTANT, CHANGED THE WAY
THEY THOUGHT ABOUT DOING TENURE
AT HARVARD.
IN 1980 IN GENERAL IF YOU WERE
IN A DEPARTMENT AS ASSISTANT
PROFESSOR THEY WOULD SAY YOU
LOOK TO YOUR LEFT AND RIGHT,
ONLY ONE OF YOU IS GOING TO BE
HERE IN A YEAR OR TWO, THAT
CHANGED DRAMATICALLY SO
ESSENTIALLY WITH FEW EXCEPTIONS
ALL ARE STILL IN THE DEPARTMENT
TODAY, SO IT'S QUITE REMARKABLE.
ONE OTHER PRINCIPLE THAT I
THOUGHT WAS REALLY QUITE -- HE
FELT YOU NEEDED COMMUNICATION,
EVEN FACE-TO-FACE COMMUNICATION,
AND THE WAY HE BROUGHT THAT
ABOUT WAS TO -- WHEN THE
DEPARTMENT ACTUALLY MOVED
SEVERAL TIMES TO DIFFERENT
BUILDINGS, AND PART OF THAT WAS
TO ENSURE THAT AS DEPARTMENT GOT
BIGGER AND MOVED THERE WAS GOOD
COMMUNICATION BETWEEN THE
FLOORS.
IF YOU LOOK AROUND HARVARD,
THERE ARE IMPRESSIVE STAIRCASES.
AS WE MOVED UP, THE STAIRCASES
GOT BIGGER AND MORE IMPRESSIVE
AND THEY ARE LITTERED THROUGHOUT
THE HARVARD CAMPUS, THAT WAS
TERRIFIC.
ANOTHER RULE PHIL HAD, THE POWER
OF HUMAN GENETICS IS REMARKABLE,
IT'S LIKE PLAYING THE LOTTERY.
SOMETIMES YOU MAP A GENE AND AS
PHIL DID HERE, HE FOUND THAT MYC
WAS TRANSLOCATED, THAT
TRANSLOCATION ACTIVATED, LEADING
TO BURKITT'S LYMPHOMA, HE FOUND
A TRANSCRIPTION FACTOR IN A
REGULATED EXPRESSION OF HIS
GENES, LIKE WINNING THE LOTTERY.
OTHERS, OTHER TIMES, I'LL SHOW
AN EXAMPLE IN OUR LAB, WHERE WE
FOLLOWED THE SAME PRINCIPLE,
MAPPED A GENETIC LOCUS, IN A WAY
NOT EXACTLY AS EXCITING AS WE
HAD THOUGHT.
SO JUST TO GO BACK, WE FIRST
STARTED, AND PART OF THIS ISSUE
OF MAKING SURE WE DON'T ALL WORK
ON THE SAME TOPIC, WE DECIDED --
KRICKET AND I DECIDED NOT TO
STAY, THE COMMUNITY IS BROAD,
INTERNATIONALLY AS WE'LL HEAR
THERE ARE MANY IMMUNOLOGISTS AND
THOUGHT WE WOULD GO IN DIFFERENT
DIRECTION, I NEEDED AN ADULT
CARDIOLOGIST, SO KRICKET
INTRODUCED ME TO A DISEASE WHICH
IN 1980 WAS STILL CALLED IHSS,
WHICH ROLLS OFF YOUR TONGUE,
MANY PEOPLE MIGHT NOT HAVE HEARD
OF IT BUT IT WAS DISCOVERED IN
THIS BUILDING OR NAMED BY GENE
BRANWALD, 1970s, A DISEASE
THAT'S BEEN AROUND FOR QUITE
SOME TIME.
YOU MAY HAVE HEARD OF IT BECAUSE
EVERY NOW AND THEN YOU HEAR
ABOUT AN ATHLETE WHO DIES
SUDDENLY ON THE FIELD, 50% WHO
DIE SUDDENLY HAVE IHSS, NO
LONGER IDIOPATHIC, BUT IN
ADDITION THERE ARE OTHER CARDIAC
DISEASES THAT CAN BE TREATED FOR
THEIR ACTUAL SYMPTOM BUT YOU
ACTUALLY UNTIL RECENTLY THERE
WAS NO TREATMENT FOR THE ACTUAL
CAUSE OF THE DISEASE.
SO, IT TURNED OUT THAT IHSS IS
INHERITED IN MANY INSTANCES
AUTOSOMAL DOMINANT TRAIT, FOUND
FAMILIES CORRECTIVELY AND
STARTED TO MAP THE GENES.
SOMETIMES YOU WOULD LAND ON THE
LOCUS OF EXCITING LIKE MYC OR
Bcl11 OR A GENE THAT MANY
WOULD CONSIDER BORING.
SO WHAT IS MYOSIN HEAVY CHAIN?
HERE YOU SEE AN iPS CELL,
GREEN, FOR THOSE WHO REMEMBER
YOUR LIGHT MICROSCOPY FROM HIGH
SCHOOL, YOU MEMBER SKELETAL
MILLION IS STRIATED MUSCLE
SARCOMERES THAT CONTRACT AND
RELAX, IT IS A COMPONENT OF THAT
CONTRACTILE APPARATUS, WE MAPPED
A NUMBER OF OTHER GENES, ALL
CAUSED MUTATIONS, EACH CAUSED
HYPERTROPHIC CARDIOMYOPATHY,
PART OF THE COMPLEX, THICK OR
THIN FILAMENT.
SO, OLD MODEL WAS THAT THE WAY
SARCOMERES WORKED, STEPPED
ALONG, THESE SMALL MOLECULES
THAT ARE GOING ON AND OFF ARE
ATP AND CALCIUM, SO BASICALLY
THE MYOSIN HEAD WAS ACTING LIKE
ATPase, IT TURNED OUT THIS IS
WHERE ONE OF PHIL'S RULES ABOUT
LAMP POST SCIENCE -- YEAH,
OKAY -- CAME INTO EFFECT.
SO HISTORICALLY TO STUDY
BIOCHEMISTS WITH CLEAVE AND
ISOLATE S1 HEAD, CONTAINED THE
MYOSIN ATPase WITH A SINGLE
ACTIVITY, THAT WAS LOOKING WHERE
WE COULD LOOK BECAUSE THEY WERE
ONLY LOOKING AT THE HEAD.
WHEN YOU CLEAVED THE MYOSIN
HEAD, THE ROD WHICH LOOKS TO BE
BORING, TURNED OUT THAT THE HEAD
ACTUALLY HAS A DOCKING SPOT,
ALONG THE MYOSIN ROD, WHEN THEY
DOCK THAT REDUCES ATPase
ACTIVITY FIVE FOLD, PRESERVING
ENERGY FOR CARDIOMYOCYTE, ONLY
ACTIVE ON THE OTHER HAND WHEN
THE HEAD IS SEPARATED FROM THE
ROD AND IS OUT BINDING THE THIN
FILAMENT.
AT ANY GIVEN TIME IN ALL OF US
90% OF OUR MYOSIN HEADS ARE
RESTING IN THE SUPER RELAXED
STATE, 10% OF TIME ARE THEY OUT
CHEWING UP ENERGY AND MAKING
MUSCLES CONTRACT.
SO, THE MUTATIONS THAT WE
IDENTIFIED WERE ONES THAT
PRIMARILY DISRUPTED THE
INTERACTION OF THE HEAD WITH THE
ROD, AND SO THEY WERE FORCING
THE MYOSIN MOLECULE CONTINUALLY
INTO THE DISORDERED STATE WHERE
IT WAS CHEWING UP ATP AND SO THE
REAL PROBLEM ASSOCIATED WITH
THIS CONDITION WAS THAT WE WERE
USING UP ATP AT A HIGHER RATE
THAN THEY NEEDED TO AND THAT
EVENTUALLY LED TO MYOCYTE DEATH
AND DYSFUNCTION.
SO, ABOUT TEN YEARS AGO
COLLECTIVELY WE COLLABORATED AND
PUT TOGETHER BY SOMEONE WHO IS
THE GODFATHER OF OUR COMPANY, WE
GOT TOGETHER, DECIDED IT WOULD
BE USEFUL TO HAVE A MOLECULE
THAT INHIBIT THE MYOSIN
ATPase, AND AFTER SCREENING A
LARGE NUMBER OF MOLECULES WE WON
THE LOTTERY, WE FOUND A MOLECULE
THAT YOU COULD TAKE IT ORALLY
AND IT DIDN'T JUST GO ANY, WHERE
JUST WENT TO THE HEART MUSCLE,
AND IN THE HEART MUSCLE WOULD
HAVE A BENEFICIAL EFFECT OF
REDUCING MYOSIN ATPase
ACTIVITY.
AND THEN AT HARVARD MEDICAL
SCHOOL IN PHIL'S MOUSE ROOM WE
COULD SHOW MICE AT HARVARD NEVER
NEED TO HAVE HYPERTROPHIC
CARDIOMYOPATHY AND DEVELOP HCM
BECAUSE WEIGH COULD GIVE THEM
MYK-461.
IF YOU WATCH CABLE NEWS, YOU
KNOW THEY FREQUENTLY ADVERTISE
MOLECULES THAT WE NEVER HEARD
OF, ONE THAT YOU MIGHT NEVER
HAVE HEARD OF IS NOW CALLED
CAMZIOS, APPROVED BY THE FDA AS
A DRUG FOR THE TREATMENT OF
HYPERTROPHIC CARDIOMYOPATHY,
CAN YOU SEE ADVERTISEMENTS ON
MSNBC.
WE BENEFITED FROM FOLLOWING
PHIL'S PRINCIPLES TAKING
ADVANTAGE OF POWER OF HUMAN
GENETICS.
I WANT TO MENTION THAT AS I SAID
IT'S NOT WHAT YOU DID FOR US
YESTERDAY, IT'S WHAT YOU'RE
GOING TO DO FOR US TODAY AND
WHAT YOU WILL DO FOR US
TOMORROW.
SO JUST QUICKLY, ONE THING THAT
STUDIES OF THE HUMAN HEART
PROGRESSED RAPIDLY BECAUSE OFFED
A VEND OF SINGLE NUCLEI RNA
SEQUENCING ANALYSIS, IT LOOKS
LIKE A SMALL THING BUT THERE WAS
A MAJOR BREAKTHROUGH WHEN RUN
RECOGNIZE IT'S YOU CAN DO THESE
BY LOOKING AT NUCLEI INSTEAD OF
WHOLE CELLS, FREEZE HUMAN TISSUE
AND JUST LOOK AT THE NUCLEUS TO
REMIND YOU CARDIAC MYOCYTE IS
100 MICRONS IN LENGTH, BIG
ENOUGH TO CLOG UP EVERYBODY'S
SINGLE CELL DEVICE BUT NOW YOU
CAN DO IT, INSTEAD OF THE SIMPLE
CARDIAC TISSUE WE THOUGHT WE
HAD, BASICALLY CARDIOMYOCYTES,
WE HAVE 11 CELL POPULATIONS,
INTERESTINGLY WOMEN HAVE MORE
CARDIOMYOCYTES THAN MEN AND THAT
MAY ACCOUNT AT LEAST IN PART FOR
WHY MEN ARE MORE SUSCEPTIBLE TO
HEART DISEASE THAN WOMEN.
MALE CARDIOMYOCYTES ARE SLIGHTLY
LARGER THAN FEMALE.
TO MAKE MATTERS EVEN WORSE, THE
DICTIONARY IS MORE COMPLICATED,
11 CELL TIMES, THEY CAN BE
BROKEN DOWN INTO SUBPOPULATIONS,
I DON'T HAVE TIME TODAY BUT YOU
CAN START TO LOOK AT THESE
SUBPOPULATIONS AND UNDERSTAND IN
VERY EXQUISITE LEVEL WHY IT
LEADS TO THICKENING OF HEART
WALL AND SHAPE IN THE LEFT
VENTRICULAR WALLS, CHAMBERS GET
LARGE, BY LOOKING AT SINGLE
NUCLEI CAN HAVE A CLEAR PICTURE
WHY THAT'S GOING ON BUT YOU HAVE
TO READ ABOUT THIS IN THE
LITERATURE, I DON'T HAVE TIME TO
DISCUSSED TODAY.
LAST THING IN LINE WITH WHAT STU
WAS TALKING ABOUT, CLEARLY SMALL
MOLECULES ARE NICE, GENE THERAPY
IS EVEN BETTER.
ONE BIG ISSUE IN THE HEART IS
HOW DID THE LIVER VECTORSES TO
TO THE HEART ALLOW SINGLE BASED
EDITING, TURNS OUT ONE NICELY
AAV GETS INTO THE MOUSE HEART,
YOU CAN PAINT THE HEART RED
USING TOMATO OR GREEN USING GFP,
GET TRANSDUCTION, USING BASE
EDITORS YOU CAN ACTUALLY TREAT
AGAIN HYPERTROPHIC
CARDIOMYOPATHY, WE'VE CHANGED
THEIR BASE OR EDITED THEIR BASES
SO THEY ARE BASICALLY CURED OR
THEY HAVE A NORMAL GENETIC
SEQUENCE.
AND THAT'S QUITE EFFICIENT,
WHETHER WE CAN TRANSLATE THAT TO
HUMANS HAS ALL THE ISSUES THAT
STU TOLD US ABOUT, DELIVERY, WE
WANT TO EDIT IN VIVO, WE DON'T
HAVE THE OPPORTUNITY OF GROWING
CELLS IN CULTURE THE WAY ONE CAN
DO IN THE HUMAN HEMATOPOIETIC
SYSTEM, SO MANY ISSUES TO GO,
THAT'S WHERE WE'RE GOING FOR THE
FUTURE.
I JUST WANT TO SAY AS I SAID NOW
IN ORDER TO AVOID THE PROBLEM OF
DOING PROBLEMS ASSOCIATED WITH
SCIENCE BY COMMITTEE WE DO
SCIENCE BY ZOOM.
THIS IS THE CURRENT ZOOM PICTURE
FROM OUR GROUP.
EVERY NOW AND THEN I LOOK AT
THIS PICTURE AND I SAY, YOU
KNOW, IT'S HARD DOING SCIENCE BY
COMMITTEE BUT SCIENCE BY ZOOM IS
A LITTLE BETTER.
THEN I WANTED TO TAKE THIS
OPPORTUNITY TO THANK THE
PATIENTS AND THE DONORS AND
FAMILIES WHO CONTRIBUTED, ALSO
THANK THE LEDER LAB AND
COMMUNITY, THANK YOU FOR THE
MANY YEARS WE ENJOYED BEING WITH
YOU AND THANK FAES FOR MAKING
THE PHIL LEDER HOUSE, A LOT OF
EDUCATION WILL COME FROM THAT
BEYOND THE 150,000 PEOPLE YOU
ALREADY EDUCATED SO THANK YOU.
[APPLAUSE]
>> NEXT SPEAKER IS PHIL HIETER,
PROFESSOR IN THE SMITH
LABORATORIES, DEPARTMENT OF
MEDICAL GENETICS, UNIVERSITY OF
BRITISH COLUMBIA, ENTERED THE
FAES JOHNS HOPKINS GRADUATE
PROGRAM, YOU MAY BE MEMBERS OF
THAT PROGRAM, AS A Ph.D.
STUDENT IN 1976, CONDUCTED
DOCTORAL RESEARCH WITH PHIL
LEDER FROM 1978 TO 1981.
HE WAS A POSTDOCTORAL FELLOW AT
STANFORD WITH RON DAVIS, JOINED
FACULTY AT JOHNS HOPKINS SCHOOL
OF MEDICINE IN LATE '80s AND
'90s, MOVED TO UNIVERSITY OF
BRITISH BRITISH COLUMBIA IN
1997, PRESIDENT OF THE GENETIC
SOCIETY OF AMERICA IN 2012 AND
PRESIDENT OF THE CANADIAN
SOCIETY FOR MOLECULAR
BIOSCIENCES IN 2017, CURRENTLY
CO-DIRECTOR OF THE CANADIAN RARE
DISEASES MODELS AND MECHANISMS
NATIONAL NETWORK.
RECOGNIZED FOR WORK ON
STRUCTURAL AND REGULATORY
PROTEINS THAT ENSURE FAITHFUL
SEGREGATION OF CHROMOSOMES IN
CELL DIVISION AND FOR RELATING
FINDINGS IN YEAST TO HUMAN
CANCER AND THERAPEUTICS.
HE'S A MEMBER OF THE NATIONAL
ACADEMY OF SCIENCES, ROYAL
SOCIETY OF CANADA.
PHIL.
>> IT'S A GREAT HONOR TO BE
HERE.
SO GREAT TO SEE THE FAMILY.
AND TO SEE MY FORMER COLLEAGUES.
A LITTLE BIT OF DEJA VU
EXPERIENCE TODAY WALKING BY
BUILDING 6 BECAUSE IT WAS IN
1975 I VISITED BUILDING 6,
INTERVIEWING WITH PHIL LEDER,
AND I WAS TOTALLY BLOWN AWAY,
MEETING WITH HIM IN HIS OFFICE,
THE INCREDIBLE ENTHUSIASM AND
PASSION THAT HE ARTICULATED, AND
JOHN SEIDMAN CAME IN WITH AN
ELECTRON MICROGRAPH OF AN
R-LOOP, DEJA VU AGAIN, IT WAS
FANTASTIC.
I WAS FORTUNATE TO BE ACCEPTED
IN THE LAB.
IT WAS REALLY SOME OF THE BEST
YEARS OF MY LIFE.
HOW DO I CHANGE THE SLIDE?
WHAT DO I DO?
WHICH ONE?
AM I IN CONTROL OF THIS?
THIS IS A PICTURE OF THE GROUP
BACK IN 1979-ISH.
AND THIS GIVES ME AN OPPORTUNITY
TO SAY -- EXPRESS GRATITUDE, IT
WAS AN INCREDIBLE EXPERIENCE.
NOT IN THE PICTURE BUT THANKS TO
JON.
JON BASICALLY WAS THE ONE WHO
TOOK ME UNDER HIS WINGS AND
TAUGHT ME CLONING AND HOW TO DO
EXPERIMENTS FAST.
AND WELL.
I REALLY APPRECIATE THAT.
THANKS TO THE WHOLE GROUP.
YOU'LL SEE FAMILIAR FACES THAT
WE'LL HEAR FROM IN A FEW
MINUTES.
I THOUGHT I WOULD JUST LIST A
FEW WHAT I CAME UP WITH, A FEW
CHARACTERISTICS OF THE AWESOME
POWER OF PHIL LEDER, FIRST I
WOULD SAY AS WE'VE HEARD BEFORE
HE KNEW HOW TO PICK IMPORTANT
RESEARCH PROBLEMS.
HE EMPHASIZED TO ME AND I THINK
EVERYBODY THE IMPORTANCE OF
TECHNOLOGY, IMPORTANCE OF
DEVELOPING NEW TECHNOLOGY,
HAVING TECHNOLOGY REALLY WORKING
IN THE LAB, SO YOU COULD
RELIABLY GET DATA AND REALLY
MAKE PROGRESS.
OF COURSE HE WAS CREATIVE AND
RIGOROUS SCIENTIST, EXUDED
PASSION AND JOY IN THE LAB, THAT
WAS JUST INFECTIOUS.
THE OTHER THING THAT FOR ME
PERSONALLY HE REALLY MADE ME
APPRECIATE THE VALUE OF
EVOLUTIONARY CONSERVATION
BECAUSE OF THE PROJECT THAT HE
GAVE ME WHEN I WAS A GRADUATE
STUDENT.
MOVING FROM HOUSE TO HUMAN
IMMUNOGLOBULIN GENES, USING
THOSE BASICALLY TO USE HUMAN
CELL LINES AND HUMAN PATIENTS
CELLS AS A MODEL FOR
UNDERSTANDING GENE
REARRANGEMENTS AND DEVELOPMENT.
AND THIS MAYBE GAVE ME THE
COURAGE TO JUMP TO YEAST AFTER
LEAVING THE LAB AND USING THAT
AS A MODEL SYSTEM FOR STUDYING
BIOLOGY RELEVANT TO HUMANS AND
HUMAN DISEASE, HUMAN BIOLOGY.
THE OTHER THING I LEARNED IS THE
POWER OF COLLABORATION AND
COLLEGIALITY, AND COMPETITION,
JOYFUL COMPETITION TO GET THINGS
DONE AND BE THE FIRST TO GET
THERE, AND HE REALLY SHOWED US
HOW COLLABORATING WITH OTHER
LABORATORIES, RESEARCH EXPERTISE
AND RESOURCES, COULD SYNERGIZE
AND DRIVE THINGS FORWARD.
AND THAT'S SOMETHING I'VE TAKEN
WITH ME THROUGH MY WHOLE CAREER.
HE ALSO INTRODUCED TO THE
CLINICIAN SCIENTIST, BASIC
SCIENTIST INTERACTION, AT LEAST
HALF THE PEOPLE IN THE LAB WERE
M.D.s, I WAS THE SOLE GRADUATE
STUDENT WITH EIGHT POSTDOCS AND
FANTASTIC CLINICIANS.
THERE WAS A VERY MEANINGFUL
CONNECTION THAT WAS MADE DURING
MY THESIS WORK.
HIS MENTORSHIP, HE WAS TRULY
ENGAGED, REALLY CARED ABOUT
PEOPLE IN THE LAB.
HE EMPOWERED US, GAVE US
OWNERSHIP OF THE WORK WE WERE
DOING TO GIVE US CONFIDENCE.
HE TAUGHT US LOTS OF THINGS, HOW
TO GIVE A GOOD TALK.
EVERYONE HAS A STORY ABOUT
LEARNING HOW TO GIVE A GOOD
TALK.
TRANSITIONS BETWEEN SLIDES AND
GROUPS OF SLIDES, FANTASTIC.
HAD HE A HUGE IMPACT ON MY
FUTURE CAREER, AND PROVIDES A
ROLE MODEL FOR ME, AND ACTUALLY
EVERYBODY SO I THOUGHT I WOULD
GO THROUGH A COUPLE THINGS,
LOOKING BACK AT THE THESIS WORK,
THE VALUE OF EVOLUTIONARY
CONSERVATION THAT STUCK WITH ME
I WOULD SAY.
AND SO PHIL'S AND JON SEIDMAN'S
GIFT TO ME IN 1978 AFTER TWO
YEARS IN BALTIMORE WAS TO
SUGGEST THE AUDACIOUS -- I DON'T
KNOW, AUDACIOUS -- THEY HAVE
ALREADY CLONE THE MOUSE GENES,
WOULD KEY CLONE THE HUMAN GENES
BY CROSS-SPECIES HYBRIDIZATION,
PROBE HEME PRE-B AND B CELL
LEUKEMIAS TO GET INFORMATION
ABOUT GENE REARRANGEMENTS AT
DIFFERENT LEVELS OF DEVELOPMENT.
IN THOSE DAYS, SEPARATED IN THE
LAB BY TWO DIMENSION METHOD,
CHROMATOGRAPHY AND FRACTIONS RUN
ON A GEL, PURIFIED BY SIZE ON
WHAT WAS CALLED THE GENE
MACHINE.
THAT ALLOWED YOU TO TAKE A
MILLION FRAGMENTS AND GET THEM
DOWN TO, SAY, THOUSANDS.
AND MADE IT POSSIBLE,
TECHNICALLY FEASIBLE, TO CLONE
GENOMIC FRAGMENTS OF DNA.
WHAT THEY HAD PREVIOUSLY SHOWN,
THREE CROSS-HYBRIDIZING
FRAGMENTS IN THE GENOME, AFTER
MUCH TIME IN A YEAR OR SO WE
FOUND THE SEQUENCE AND IT WAS A
REPEAT IN THE FRAGMENT OF THE
MOUSE GENE, VERY DISAPPOINTING
BUT, AGAIN, I MEAN IN TYPICAL
PHIL LEDER FASHION HE WAS LET'S
GET RIGHT BACK TO IT AND FIGURE
THIS OUT.
SO, THE SOLUTION WAS TO RESTRICT
THE PROBE TO THE OPEN READING
FRAME AND THAT SHOWED THE 2 1/2
KB FRAGMENT WAS THE KEY.
SO, THAT LED TO CLONING OF THE
HUMAN KAPPA GENE.
THIS PARTICULAR FIGURE STICKS IN
MY MIND, WHERE IT'S A
HETERODUPLEX BETWEEN -- I WON'T
TRY TO POINT ON THIS, BUT
BETWEEN THE HUMAN KAPPA GENE
WITH J REGION GENES IN THE MOUSE
GENE, YOU CAN SEE HOMOLOGY
SEGMENTS THAT CORRESPOND TO THE
C REGION, CONSTANT REGION,
CONSERVED SEQUENCE IN
INTERVENING SEQUENCE, EACH OF
THE J REGIONS FORMING
HETERODUPLEX, SO YOU CAN MAP
WHERE ON THESE CLONES THESE
ELEMENTS ARE, AND THE SEQUENCE
WAS VERY REVEALING, SHOWED
EVOLUTIONARY CONSERVED NOT ONLY
AT THE AMINO ACID LEVEL BUT
SEQUENCES IMPORTANT FOR VJ
RECOMBINATION AND PROBABLY
SPLICING.
ONCE WE SUCCESSFULLY CLONED THE
IMMUNOGLOBULIN KAPPA GENE WERE
ABLE TO GET HUMAN LAMBDA GENES,
ARRANGED IN A TANDEM, EACH WITH
A SINGLE J REGION.
AND THEN WITH THOSE CLONES IN
HAND THIS GETS TO THE POWER OF
COLLABORATION AND THE LINKAGE
BETWEEN CLINICIAN AND BASIC
SCIENTIST.
PHIL AFTER CLONING THE KAPPA
GENE BASICALLY SET UP AN
APPOINTMENT WITH TOM WALDMAN, AN
IMMUNOLOGIST AT -- WAS AT NIH,
AND I REMEMBER HIM TAKING ME
ACROSS CAMPUS AND SITTING IN THE
OFFICE AS A GRADUATE STUDENT
WITH PHIL AND TOM WALDMAN, AND
HE EXPLAINED WE WERE CLONING
THESE GENES, WANTED TO USE THEM
AT PROBES TO LOOK AT B CELL
DEVELOPMENT STAGES.
HE WANTED ME TO HOOK UP WITH A
RESEARCH FELLOW THAT'S IN HIS
GROUP AND WANTED TO BE THE BEST
ONE, THAT'S WHAT HE TOLD ME.
OKAY, WHO WAS IT?
STAN KORSMEYER, QUITE AN AMAZING
GUY.
HE WAS A FELLOW IN WALDMAN'S
GROUP, AND WE ESTABLISHED A
COLLABORATION WHERE HE HAD THE
SORT OF CELL LINES AND HAD
PATIENTS WITH DIFFERENT TYPES OF
KINDS OF LEUKEMIA, AND THOSE
WERE THE CELLS WE USED TO LOOK
AT GENE REARRANGEMENTS AT
DIFFERENT STAGES OF DEVELOPMENT.
BASICALLY WE SHOWED A HIERARCHY
IN THE GENE REARRANGEMENT
PROGRAM, SO THE MU HEAVY CHAIN
WOULD REARRANGE FIRST FOLLOWED
BY KAPPA REGION, LAMBDA GENE,
SOME KIND OF FEEDBACK WHEN THERE
WAS A FUNCTIONAL REARRANGEMENT,
THE PROCESS STOPPED.
SO, THIS WAS KIND OF EXPLAINED,
EXCLUSION WERE BIG ISSUES, VERY
SATISFYING.
STAND WENT ON TO DO TERRIFIC
WORK IN HIS OWN RIGHT AND
UNFORTUNATELY PASSED AWAY AT A
YOUNG AGE.
AND THEN THE LAST PIECE IS THE
MENTORSHIP PIECE.
AS I SAID, HE WAS JUST AN
INCREDIBLE MENTOR, AND ONE OF
THE THINGS I REMEMBER HIM DOING
ABOUT A YEAR BEFORE I WAS
FINISHED IN THE LAB WAS TO COME
TO ME IN THE HALLWAY AND SAY YOU
NEED TO THINK ABOUT WHAT YOU
WANT TO DO, WHAT'S YOUR NEXT
PHASE, WHERE DO YOU WANT TO DO A
POSTDOC?
AND HE SAID, GO AWAY FOR A
MONTH, AND TALK TO AS MANY
PEOPLE AS YOU CAN.
COME BACK AND LET'S, YOU KNOW,
DECIDE WHERE YOU WANT TO GO AND
LET'S GO FOR IT.
I DID THAT.
I TALKED TO ALL THESE PEOPLE IN
THE LMG.
I REMEMBER REALLY GREAT
DISCUSSIONS WITH BOB CROUCH,
MIKE CASHELL AND OTHERS ON THE
FLOOR, THEY INTRODUCED ME TO THE
WORK OF RON DAVIS WHICH I DID
NOT KNOW ABOUT.
HE HAD IDENTIFIED YEAST DATA,
REPLICATION, INVOLVED IN
DEVELOPING A LOT OF CLONING
TECHNIQUES THAT WE WERE USING IN
THE LAB.
AND THIS SEEMED LIKE -- AND THEY
WERE WORKING IN YEAST.
I THOUGHT, PHIL SAID LET'S DO
SOMETHING TOTALLY DIFFERENT AND
TRY TO COMBINE THEM INTO YOUR
FUTURE LAB.
SO I WAS -- THIS WAS TAKING AN
EVOLUTIONARY JUMP FROM THE
MAMMALIAN MODELS TO YEAST AS A
MODEL, OVER A BILLION YEARS OF
EVOLUTION AS A MODEL SYSTEM FOR
STUDYING BASIC EUKARYOTIC
BIOLOGY.
AND THAT WAS AROUND THE EARLY
'80s, THAT WAS TIME WHEN DNA
MEDIATED TRANSFORMATION HAD JUST
BEEN WORKED OUT AND WAS SEEN
THAT DNA ACTUALLY BEHAVED IN
HOMOLOGOUS REGIONS IN THE
CHROMOSOME AND WE CAN DO GENE
REPLACEMENTS IN THE GENOME IN
THE EARLY '80s, AT THE SAME
TIME CENTROMERES AND ORIGINS
REPLICATION WERE BEING DEFINED
THAT ALLOWED DEVELOPMENT AT THE
SOMAL VECTORS FOR INTRODUCING
GENES IN THE YEAST.
THIS WAS A VERY POWERFUL SYSTEM,
AWESOME POWER OF YEAST GENETICS,
AND THE OTHER THING I'LL MENTION
IS REALLY BY THE END OF THE
'80s THERE WAS THIS PUSH TO
SEQUENCE THE HUMAN GENOME,
RECOGNIZED BY A PANEL RUN BY THE
NRC, THE COMMITTEE ON HUMAN
GENOME PROJECT WHICH RELEASED
REPORT IN 1988 THAT REALLY THE
GENOME PROJECT NEEDED TO INCLUDE
THE HUMAN GENOME AND MODELS
SYSTEMS, YEAST, FLIES, MICE.
ON THAT COMMITTEE, STU ORKIN AND
SHIRLEY TILGHMAN, I WAS GOING TO
THANK THEM FOR THAT BRILLIANT
ARCHITECTS REALIZED WE NEEDED
MODEL SYSTEMS TO FUNCTIONALIZE
THE GENES.
AND SO VERY QUICKLY, I'M NOT
GOING TO DESCRIBE ALL THE WORK
WE'VE DONE BUT WE DECIDED THE
PROBLEM WAS TO USE YEAST AS A
MODEL FOR STUDYING MECHANISMS OF
CHROMOSOME SEGREGATION,
STABILITY, THEREFORE CANCER
BIOLOGY BECAUSE IT WAS
RECOGNIZED THAT TUMORS HAVE
PROPERTY OF GENOME INSTABILITY,
THIS DRIVES THE EVOLUTION OF
PROGENITOR CELLS SO THEY CAN
ACCUMULATE ALL OF THE GENES
REQUIRED FOR THE CANCER CELL
PHENOTYPE.
BUT WE COULD USE THE YEAST
SYSTEM TO DEFINE THE GENES,
STUDY BIOLOGY AND BY DEFINITION
THESE REPRESENTED CROSS-SPECIES
CANDIDATE GENES THAT COULD BE
MUTATED IN TUMORS AND THEN GO ON
TO USE YEAST AS A GENETIC SYSTEM
TO CONSIDER THESE GENES, THESE
MUTATIONS AS GENETIC
VULNERABILITIES THROUGH THE
CONCEPT OF SYNTHETICICALLY
VALIDATED FINDING GENES THAT
WOULD SELECTIVELY KILL A CANCER
MUTANT, DEFECT DEFECTIVE IN
CHROMOSOME SEGREGATION.
IN 1985 ONE THING PHIL DID WAS
GIVE THEM COURAGE TO TACKLE BIG
PROBLEMS, REQUIRING NEW
TECHNOLOGIES.
THIS COMES FROM TRAINING FROM
JON SEIDMAN AND OTHERS IN THE
LAB WAS TO DEVELOP A SYSTEM
WHERE WE COULD GENERATE
ARTIFICIAL CHROMOSOMES, EXTRA
CHROMOSOMES, AS MARKERS TO
MONITOR THE STABILITY OF
CHROMOSOME SEGREGATION, IN A
SECTOR ASSAY, CARRYING A COLONY
COLORED MARKER WHICH GAVE RISE
TO RED SECTORS ON CHROMOSOME
LOSS AT RARE FREQUENCY, IDENTIFY
MUTANTS THAT INCREASE FREQUENCY
OF SEGREGATION AND USE AS POINT
FOR CLONING BY COMPLEMENTATION,
YOU SEE AN EXPERIMENT WHERE
GENOMIC LIBRARY WAS PLACED IN A
MUTANT, THERE'S A COMPLEMENTED
CLONE, WE CAN RESCUE THE PLASMID
AND FIND THE GENE.
THERE WAS A LOT OF -- ONE AREA
WAS THE YEAST GENETIC CORE, MANY
CTF MUTANTS WERE INVOLVED IN
FUNCTION, ANOTHER CLASS INVOLVED
SISTER CHROMATID COHESION,
CENTROMERE FUNCTION AND COHESION
ARE INTIMATELY INTERTWINED IN
TERMS OF FUNCTION BECAUSE THAT
ALLOWS THE CELL TO ACHIEVE
BIPOLAR ATTACHMENT OF
CHROMOSOMES TO ACHIEVE META
PHASE, THERE'S A SIGNAL
TRANSDUCTION MECHANISM THROUGH
THE SPINDLE CHECKPOINT THAT
REGULATES THE META
PHASE/ANAPHASE TRANSMISSION,
INVOLVING DEGRADATION OF KEY
CYCLE REGULATORS.
SO THE STORY THAT GETS US BACK
TO PHIL WAS IN 1992, I WAS THE
FIRST -- BEGINNING TO CLONE
HUMAN ORTHOLOGS OF YEAST GENES,
STUDYING YEAST, OF COURSE THERE
WAS NO HUMAN GENE, NO
ORTHOLOGS.
A STUDENT STARTED TO DO THE
ORTHOLOGS, IN A RESTED META
PHASE, A CHECKPOINT THAT
MONITORINGS DNA DAMAGE, WE
THOUGHT THESE WERE COMPONENTS OF
THE SPINDLE.
AND HE TRIED ALL WAYS TO CLONE
THESE THINGS BY FUNCTIONAL
COMPLEMENTATION, AND OTHER
METHODS TO GENERATE PCR, IT WAS
UNSUCCESSFUL.
BUT WHAT THE BREAKTHROUGH CAME
WHEN THE FIRST ROUND OF
EXPRESSED SEQUENCE TAGS WERE
RELEASED PUBLICLY, 8,000,
ORIGINALLY FOUND AT NIH BY CRAIG
BENTNER, ACATCHED TO A CLONE IN
THE FREEZER, A WEAK HOMOLOGY
BETWEEN CDC27 AND OPEN READING
FRAME ON THE EST.
WE BEING AT HOPKINS IN THE
MEDICAL GENETICS COMMUNITY, WE
MAPPED THIS CLONE, FULL LENGTH
COPY WITH 20% IDENTITY OF THE
AMINO ACID LEVEL, AN ORTHOLOGUE,
IT MAPPED AND WAS IN THE BRCA1
800 KB SEGMENT, BECAME A
CANDIDATE GENE FOR A MONTH, WE
SENT PROBES.
THIS HAD A HUGE IMPRESSION ON
ME.
WHAT IF WE DID THIS
SYSTEMATICALLY AND IDENTIFIED
ESTs RELATED TO PROTEINS BEING
ACTIVELY STUDIED IN YEAST WITH
ANNOTATE MAPS WITH BIOLOGY OF
YEAST AS CANDIDATE GENES, AND
THE STORY I'LL TELL, IT'S A
LITTLE OVERKILL HERE, BUT IN
1992 I WAS RECRUITED TO BE ON
THE GENOME COMMITTEE, SO I HAD
TO GO TO THE MEETINGS.
I SUBMITTED AN ABSTRACT, AND IT
WAS REJECTED.
THE ABSTRACT WAS REJECTED.
I WENT TO THE MEETING.
I DON'T THINK IT WAS A
REFLECTION OF THE HUMAN
GENETICIST VIEW AS A MODEL
SYSTEM BUT THERE WAS SKEPTICISM
ABOUT THE VALUE OF YEAST AT THE
TIME, I WOULD SAY.
WE GOT A GRANT TO DO THIS
SYSTEMATICALLY AND WE MAPPED 300
YEAST GENE EST-RELATED CLONES TO
THE HUMAN MAP AND WHICH BRINGS
US BACK TO PHIL THAT THIS WAS
ONE OF THE MOST GRATIFYING TALKS
I'VE GIVEN IN MY CAREER,
AMERICAN SOCIETY OF HUMAN
GENETICS HAD A SESSION ON
LESSONS OF MODEL ORGANISM TO
HUMAN DISEASE.
PHIL LEDER WAS AWARDED THE
ALLLAN AWARD AND GAVE A TALK TO
THE GENETIC APPROACH TO CANCER
PROBLEM, MY TALK WAS IMMEDIATELY
AFTER HIS, I WAS ABLE TO
PUBLICLY THANK HIM WITH HIM IN
THE ROOM, VERY SPECIAL, TO ME,
HERE MY SECOND CHANCE TO THANK
HIM, IN FRONT OF HIS FAMILY, SO
THAT'S REALLY GREAT.
ONE THING I WOULD LIKE TO SAY
WE, IN CANADA, AS YOU KNOW AFTER
2010, WHEN IT DROPPED FOUR
ORDERS OF MAGNITUDE SCIENTISTS
IDENTIFIED RARE DISEASE GENES
THROUGH WHOLE EXOME OR WHOLE
GENOME SEQUENCING, AND OVER THE
LAST TEN YEARS THERE'S BEEN A
RARE DISEASE GENE DISCOVERED PER
DAY, OVER 300 PER DAY FOR THE
LAST TEN YEARS, CANADIAN
SCIENTISTS IDENTIFIED 700 RARE
DISEASE GENES IN THE LAST TEN
YEARS, INITIATING A NETWORK THAT
ESSENTIALLY WAS A DATING
SERVICE, WHEN A CLINICIAN
SCIENTISTS FINDS A RARE GENE WE
CREATE A DATABASE, WE TRY TO
CONNECT THEM AT THE TIME OF GENE
DISCOVERY, SEED FUNDING MONEY TO
ESTABLISH A COLLABORATION, AND
THAT HAS BEEN VERY PRODUCTIVE
EXERCISE AND WE HAVE FUNDING TO
2027.
LET ME END BY SUMMARIZING WHAT I
THINK PHIL'S LEGACY HAS BEEN TO
ME AND SO MANY PEOPLE.
HIS SCIENCE, FIRST OF ALL,
PICKING AN IMPORTANT PROBLEM,
IMPORTANCE OF TECHNOLOGY, TO
ATTACK THOSE PROBLEMS.
HIS CRITICAL THINKING,
INCREDIBLE TALENT IN SCIENCE
THAT WE ALL TRY TO EMULATE, THE
SECOND IS HIS MENTORSHIP AND,
AGAIN, HE WAS JUST SUCH A CARING
AMAZING MENTOR, EXUDED PASSION
AND OPTIMISM, HE WAS JUST A
WONDERFUL PERSON.
IT WAS JUST WONDERFUL TO HEAR
HIM TALK.
I DON'T KNOW IF YOU'VE LISTENED
TO OR LOOKED UP HIS INTERVIEWS
WHEN HE WON THE -- FROM THE
ALBERT LASKER FOUNDATION, BUT
THEY ARE AMAZING.
I'D RECOMMEND YOU CHECK IT OUT.
AND AS A ROLE MODEL.
AND I THINK, YOU KNOW, I'VE
STRIVED TO EMULATE PHIL IN MY
OWN CAREER AND NEVER COULD QUITE
REACH THAT LEVEL BUT I WANTED TO
SHOW THIS PICTURE BECAUSE WE GO
BACK TO THE LAB OF ABOUT TEN
PEOPLE IN 1979, THAT'S MY FIRST
LAB AT HOPKINS, I'VE ACTUALLY
MAINTAINED -- I THINK IT'S A
MAGIC NUMBER, TEN PEOPLE MY
ENTIRE CAREER.
SO OVER THE YEARS THIS IS MY
CREW NOW, THERE'S ABOUT TEN
PEOPLE.
32 PhDs GRADUATED, 16
POSTDOCS, AND I SEE A DIRECT
CONNECTION BETWEEN PHIL'S
MENTORSHIP OF ME AND WHAT HE
GAVE TO ME, AND ALL THESE PEOPLE
THAT I'VE TRAINED OVER THE
YEARS.
SO THANK YOU VERY MUCH.
[APPLAUSE]
>> WOW!
NEXT SPEAKER IS JEFF RAVETCH,
PROFESSOR AT ROCKEFELLER
UNIVERSITY, HEAD OF THE WAGNER
LABORATORY OF MOLECULAR GENETICS
AND IMMUNOLOGY, UNDERGRADUATE AT
YALE UNIVERSITY, RECEIVED MD/PHD
FROM CORNELL MEDICAL SCHOOL, GOT
HIS Ph.D. IN 1978 IN GENETICS,
POSTDOC AT THE NIH WITH PHIL,
IDENTIFIED AND CHARACTERIZED
GENES FOR HUMAN ANTIBODIES AND
DNA ELEMENTS INVOLVED IN SWITCH
RECOMBINATION.
FROM 1982 TO 1996 A MEMBER OF
THE FACULTY AT SLOAN-KETTERING
AND CORNELL, LABORATORY
ESTABLISHED FC DOMAIN FUNCTIONAL
DIVERSITY AND PREEMINENCE OF FCR
PATHWAYS IN HOST DEFENSE
INFLAMMATION AND TOLERANCE.
A MEMBER OF THE NATIONAL
ACADEMIES OF SCIENCE AND
MEDICINE, AMERICAN ACADEMY OF
ARTS AND SCIENCES.
HIS WORK HAS BEEN RECOGNIZED BY
NUMEROUS AWARDS, INCLUDING PEW
SCHOLAR, WOLF PRIZE IN MEDICINE,
AND MOST RECENTLY I THINK OF
INTEREST TO THIS AUDIENCE THE
WILLIAM POHL MEMORIAL AWARD.
JEFF?
>> THANKS.
>> THANK YOU.
IT'S GREAT TO BE BACK AND TO SEE
OLD FACES.
WE HAVE THE SAME SLIDES, OUR
IMAGINATIONS ARE LIMITED BY WHAT
PHIL LEFT US.
THIS REMIND ME OF WONDERFUL
ANECDOTES WE CAN TALK ABOUT AT
LUNCH.
I CAN'T RESIST THE IDEA LOOKING
AT PHIL HIETER NEXT TO PHIL
LEDER, LADY DI WAS IN OUR
THOUGHTS AND YOU MAY RECALL HIM
DRESSING UP AT LADY DI FOR
HALLOWEEN.
1979, LATE SPRING, GIVEN A CART
TO DO MY WORK INSTEAD OF A DESK.
IT WAS PHIL'S WAY OF TESTING MY
RESOLVE, WOULD I SURVIVE?
I PROCEEDED TO PARK MYSELF AT
ANY BENCH TEMPORARILY VACATED,
DISPLACING THE LEGITIMATE
OCCUPANT IN THE PROCESS.
AFTER SEVERAL MONTHS OF THIS
ITINERANT LIFESTYLE I ACQUIRED A
STABLE SPOT, BIGGER THAN AN ICE
BUCK ET WITH A POWER OUTLED.
IT WAS EXCITING.
THERE WERE NEVER ENOUGH GEL
BOXES, POWER SUPPLIES, WATER
BATHS, ENZYMES, ALL THE REAGENTS
WE NEEDED TO CLONE AND SEQUENCE
GENES, IN A RACE TO CHARACTERIZE
HEAVY GENES, PHIL WOULD LOOK
OVER AS THE SEQUENCING GELS CAME
OUT, EAGER TO SEE WHAT WE FOUND.
WE CRAMMED OURSELVES INTO THE
LAB DURING THE DAY, WORKED IN
SHIFTS NIGHTS AND WEEKENDS TO
COMPLETE THE WORK IN RECORD TIME
TAKING TURNS, WATCHING THE
LIBRARY SCREENS, SETTING UP
HYBRIDIZATIONS, INOCULATING
CULTURES TO GROW.
IN THE BACKGROUND WE LISTENED TO
THE RADIO FAVORING CLASSICAL
MUSIC, PHIL WAS A CON STAND
PRESENCE, STOPPING AT OUR
BENCHES WHEN DATA WAS COMING.
WE WORKED TO CLONE cDNA IT'S
AND GERMLINE REARRANGE FOR EACH
OF THE HEAVY CHAINS USING
CROSS-SPECIES HOMOLOGY WHEN WE
COULD.
PHIL WOULD LISTEN AND RETURN
LATER REPEATING RESULTS IN A
POLISHED VERSION.
I WAS BEWILDERED UNTIL I
RELATESSED HE WAS READY TO
LAUNCH INTO A MANUSCRIPT FOR A
TALK.
HE TAUGHT US RULES, NEVER
PRESENT A DATA SLIDE WITHOUT
PREPARING THE GROUND WORK, MAKE
THE AUDIENCE THINK THEY REACHED
THE CONCLUSION ON THEIR OWN
WITHOUT BEING TOLD.
AND LESS IS ALWAYS MORE.
WE TEAMED UP TO CHARACTERIZE MU
AND ALPHA HEAVY CHAINS, CLONED,
IN THE FIRST PAPER -- NOW I'M
TRYING TO FIGURE IT OUT.
WHAT'S THE SECRET?
THERE YOU GO.
GOT IT.
SO FIRST PAPER PUBLISHED IN
PNAS, WE USED HETERODUPLEX
MAPPING TO DEFINE RELEVANT
REGIONS OF THE MU AND ALPHA
GENES WITHOUT HAVING TO SEQUENCE
THEM.
IT'S STILL A FAVORITE BRINGING
BACK MEMORIES OF NIGHT IN THE
BASEMENT OF BUILDING 6 STARING
AT EM GRADES.
A PRESCIENT STUDY DEFINING
SWITCH REGIONS WITH HOMOLOGY AND
SWITCH RECOMBINATION, NOT TOO
SHABBY.
NEXT CAME DEMAND FOR SEQUENCING
GEL BOXES, PLATES, POWER
SUPPLIES AND FREEZER SPACE, TO
SEQUENCE I DECIDED TO START MY
EXPERIMENT ALSO ON A CHRISTMAS
EVE AND WORK THROUGH THE HOLIDAY
TAKING OVER GEL BOX, POWER
SUPPLY AND BENCH IN THE EMPTY
LAB, AS SOON AS I MOVED FROM ONE
END TO THE OTHER IT WAS TIME TO
START ALL OVER WITH A SECOND AND
THIRD LOAD UNTIL I REACHED MAX
UNIT, THE TERM WE COINED TO
ACKNOWLEDGE 5.5 KILOBASES AND
TOLD BY PHIL IT WAS GOOD ENOUGH,
AND PUT THE DATA TOGETHER
RESULTING IN A CELL PAPER THAT
DEFINED THE LOCUS.
NEXT SLIDE PLEASE.
THANK YOU.
OTHER PAPERS FOLLOWED WHICH STAN
KORSMEYER AND OTHERS.
SO MY THREE YEARS IN HIS LAB
WITH ONE OF THE MOST EXCITING
TIMES IN MY CAREER.
THE FOREFRONT OF IMMUNOLOGY,
USING CUTTING EDGE EXPERIMENTS,
YIELDING UNEXPECTED RESULT,
GIVING ME A TASTE FOR WHAT I
WANTED TO DO FOR THE NEXT 40
YEARS APPLYING TOOLS TO PROBLEMS
IN THE IMMUNOLOGY.
I RECALL THE DAY I SAT IN HIS
OFFICE THREE YEARS LATER, TIME
TO SET OFF ON MY OWN, LAB MOVING
TO BOSTON, I WAS GOING HALFWAY
BACK TO NEW YORK WHERE I DID MY
GRADUATE STUDIES.
PHIL ASKED WHAT I WAS PLANNING
ON WORK ON.
I SAID I THINK I'D LIKE TO WORK
ON SWITCH RECOMBINATION.
PHIL SAID NOTHING FOR WHAT SEEMS
LIKE MINUTES.
AND THEN SAID I WOULDN'T
RECOMMEND THAT.
FIND SOMETHING NO ONE IS WORKING
ON.
PROBABLY THE BEST ADVICE I'VE
EVER RECEIVED.
FORCED KNEE -- ME INTO A
PROBLEM THAT OCCUPIED ME FOR THE
REST OF MY CAREER.
NO ONE CARED ABOUT THE FC, THEY
ASSUMED IT WAS ACCOUNTED FOR BY
THE CLASSICAL PATHWAY OF
CONFLICT ACTIVATION, FC
RECEPTORS NOT ON ANYONE'S RADAR.
I TOOK MY FREEZER BOXES AND
PACKAGING EXTRACTS AND GENES I
CLONED, TWO WEEK OLD SON, HEADED
BACK TO NEW YORK.
NEXT SLIDE PLEASE.
TO START MY LAB AT
SLOAN-KETTERING.
THIS IS MY ATTEMPT TO CHANNEL MY
INNER FORESTER.
NEXT SLIDE.
SLOAN-KETTERING IN THE
BACKGROUND, ROCKEFELLER
UNIVERSITY IN THE FOREGROUND,
CORNELL MEDICAL ON THE
RIGHT-HAND SIDE OF THE SLIDE.
IT SET ME ON THE PATH PHIL HAD
SHOWN ME.
WE DEFINED THE cDNAS AFTER
YEARS OF EFFORT USING TRICKS
PHIL TAUGHT US AND NEXT SLIDE
WERE ABLE TO DEFINE COMPLEX
FAMILY OF FC RECEPTORS THAT
MODULATED BIOLOGICAL ACTIVITY OF
ANTIBODIES IN A VARIETY OF
APPLICATIONS.
SO GOING INTO THE DETAILS OF
THESE FC RECEPTORS, MEMBERS OF
THE IG SUPER FAMILY ON ONE CASE,
FUNCTION TO COUPLE THE
RECOGNITION DOMAINS OF THE FAB
MEDIATING POSITIVE AND NEGATIVE
SIGNALING PATHWAYS, WE KNOCKED
OUT THE GENES IN VARIOUS MOUSE
MODELS, AND DEFINED HOW THEY
IMPACTED ON THE BIOLOGY OF THE
SYSTEM AND THEN DID THE
COORDINATE MAPPING OF FC REGIONS
TO DETERMINE WHERE ENGAGEMENT
WOULD OCCUR, LEADING TO ABILITY
TO MANIPULATE THE FC REGION TO
DEVELOP NOVEL THERAPEUTICS AND
THIS IS A LIST OF SOME HALF
DOZEN ANTIBODIES THAT HAVE BEEN
FDA APPROVED WHOSE FCs WERE
MODIFIED TO SELECTIVELY ENGAGE
SPECIFIC FC RECEPTORS TO ENHANCE
OR MODULATE THE BIOLOGICAL
ACTIVITY OF ANTITUMOR ANTIBODIES
AS WELL AS ANTI-INFLAMMATORY
ANTIBODIES.
SO OVER THE YEARS WE'VE FOCUSED
ON HOW WE CAN COUPLE THIS SYSTEM
TO A VARIETY OF PATHWAYS, NEXT
SLIDE, YOU CAN SEE THE CURRENT
EXCITEMENT IN THE FIELD OF
MEDICAL ONCOLOGY, USING
ANTIBODIES TO BLOCK VARIOUS
CHECKPOINTS ON T CELLS, OUR
FOCUS HAS BEEN ON THE RIGHT-HAND
SIDE HOW WE CAN ACTIVATE APC BY
USING MOLECULE CALLED CD40, WELL
KNOWN TO THE IMMUNOLOGICAL
COMMUNITY, ALWAYS USED WHEN WE
WANT TO ENHANCE BIOLOGICAL
RESPONSE IN AN ANIMAL.
BUT WHEN ATTEMPTS WERE MADE,
NEXT SLIDE, TO USE THE ANTI-CD40
APPROACH AS THERAPEUTIC, NEXT
SLIDE PLEASE, IT WAS FOUND THAT
THE ANTI-C 40 AGONISTIC
ANTIBODIES WERE HIGHLY TOXIC AND
GAVE ESSENTIALLY NO IN VIVO
BIOLOGICAL RESPONSE, A SHOCK.
GOING BACK TO THE WAY PHIL
TAUGHT US TO THINK ABOUT THESE
TYPES OF PROBLEMS, IT MEANT THAT
OUR CROSS-SPECIES MODELS WERE
INADEQUATE, NOT ACCOMMODATING
THE SYSTEMS WHERE THE ANTIBODIES
WERE FIRST DEVELOPED AND
TRANSLATION INTO THE CLINICAL
ASPECT.
SO WE DID THAT BILLING
BILLING -- BUILDING MOUSE
MODELS, NEXT SLIDE, WE
APPRECIATED A SIMPLE NOTION THAT
FOR MEMBERS OF THE SUPER FAMILY
OF WHICH CD40 IS ONE THE
INTERACTION OF LIGAND IS A
TRIMER-TRIMER INTERACTION.
ANTIBODIES DIMERIZE AND
THEREFORE WERE VERY INEFFICIENT
WHEN IT CAME TO STIMULATING AN
AGONISTIC RESPONSE ON DENDRITIC
CELL, WE CAN USE THE FC AS A
SCAFFOLD TO CROSS-LINK IN
TRENDS, THE WAY IS TO USE AN FC
RECEPTOR, BUT WILL ACT INERTLY
SO WE REENGINEERED AN EXISTING
THERAPEUTIC ANTIBODY IN SEVEN
PHASE 1 STUDIES.
NEXT SLIDE.
DEMONSTRATING WE COULD ENHANCE
THE IN VIVO ACTIVITY QUITE
DRAMATICALLY.
YOU CAN SEE IF YOU COMPARE THE
PARENT ANTIBODY WHICH IS SHOWN
IN GREEN TO THE FC ENHANCED
VERSION OF THE SAME FAB IN
PURPLE WE CAN SHOW COMPLETE
ERADICATION OF MG 38 AND
METASTATIC MELANOMA MODEL IN THE
LUNG.
SO WE WERE FACED WITH A DILEMMA,
I COULD SATISFY MY WIFE'S
CRITERIA, WHAT DOES YOUR HUSBAND
DO, HER ANSWER WAS IF YOU WERE A
MOUSE HE COULD CURE YOU.
WE WANT TO GO BEYOND THAT, COULD
WE DO THIS IN A CLINICAL
SETTING?
THIS WAS A SYSTEM DEMONSTRATED
MULTIPLE TIMES, FAILED IN THE
CLINIC, NOBODY WANTED TO TOUCH
IT.
AT ROCKEFELLER WE HAD INSIGHTFUL
TRUSSEES INCLUDING JULIAN
ROBINSON, WHEN I PRESENTED THIS,
USING A REENGINEERED ANTIBODY HE
SAID I CAN WRITE A CHECK, AND HE
DID, USING PHILANTHROPIC FUNDING
WE WERE ABLE TO GMP MANUFACTURE
AND DO FIRST IN HUMAN STUDIES OF
FC ENGINEERED AGONISTIC
ANTI-CD40 ANTIBODY.
IT HAS HIGH SYSTEMIC TOXICITY.
WE CONFIRMED WHEN WE
REENGINEERED TO INCREASE
POTENCY, BUT WHY GIVE
SYSTEMICALLY WHEN YOU TRY TO
ACTIVATE IMMUNE RESPONSE IN THE
HUMAN MICROENVIRONMENT TO THEN
GIVE RISE TO SYSTEMIC
DIVERSIFICATION?
THAT'S WHAT VACCINATION DOES.
WE DECIDED TO ACTIVATE TUMOR
LOCALLY AND COUNT ON THE FACT
THAT THE ACTIVATED CD8 CELLS
WOULD THEN MIGRATE TO LYMPH
NODES AND DISPERSE THROUGH
ORGANISM TO ERADICATE OTHER
TUMORS.
DOES IT WORK?
YEAH, AMAZING.
MELANOMA IN TRANSIT, TEN YEARS
AGO, CAME BACK TEN YEARS LATER,
SLOAN-KETTERING, DIFFUSE
METASTATIC MELANOMA, YOU CAN SEE
BOTH DARK SPOTS AS WELL AS
LESIONS ON THE FOOT DOWN TO THE
FOOT.
WE DECIDED TO INJECT INTO THAT
ONE LESION A 2-MILLIGRAM DOSE,
AFTER MULTIPLE CYCLES OF
TREATMENT YOU CAN SEE PARTIAL
RESPONSE BY SIX CYCLES, AND BY
THE 15th CYCLE THERE WAS
NOTHING LEFT TO INJECT.
WE SAW RESIDUAL REGIONS OF
MELANOTIC FIGMENTATION, NO
TUMOR, COMPLETE RESPONSE.
WASN'T THE ONLY ONE.
NEXT SLIDE SHOWS EXAMPLE OF WHAT
HAPPENED IN A PATIENT, NEXT
SLIDE PLEASE, WHO HAD RECURRENCE
OF METASTATIC BREAST CARCINOMA,
THIS IS AN ESTROGEN RECEPTOR
POSITIVE HER2-NEGATIVE
METASTATIC BREAST CANCER,
MULTIPLE ROUND OF CHEMOTHERAPY,
RADIATION, OF COURSE SURGERY,
AND PRESENTED ONCE AGAIN WITH
MULTIPLE LESIONS IN HER AXILLA
AND SHOULDER, AND DEEP LIVER
METASTASES, WE INJECTED IN THE
TWO SITES WITH BLUE ARROWS,
LOCAL INJECTION, 5 MILLIGRAMS AT
EACH SITE.
YOU SEE IMMEDIATE RESPONSE, BY
SECOND CYCLE LESIONS STARTING TO
LEVEL OUT, BY FOURTH CYCLE
NOTHING LEFT TO INJECT,
COMPLETELY RESOLVED AS WELL AS
DEEP LIVER METASTASIS.
WHAT'S GOING ON?
NEXT SLIDE.
WHEN YOU BIOPSY THE SITES OF
NON-INJECTED TUMORS FROM BOTH
PATIENTS, WHAT YOU SEE ARE NOT
TUMOR CELLS, YOU INSTEAD SEE
INFILTRATION OF LYMPHOID CELLS,
THEY HAVE WELL ORGANIZED T CELL
ZONES, B CELL ZONES, DENDRITIC
CELLS, CANONICAL DENDRITIC
CELLS.
WHEN WE INTRODUCE TREATMENT YOU
SEE IMMEDIATE DECLINE IN THE
LEVEL OF TUMOR BURDEN IN THIS
PATIENT DEMONSTRATING IN FACT
INTRATUMOR EFFECTS CAN BE
MOBILIZED, IT'S NOT ONLY
RECOGNITION OF FAB ENGAGING
TARGET ANTIGEN BUT CAN PLAY
DRAMATIC ROLE IN EFFICACY.
LET ME END WITH AN ILLUSTRATION
I FOUND A LECTURE GIVEN IN 1900
FOLLOWING ON THE WORK OF THE
DESCRIPTION OF ANTI-TOXINS AS A
THERAPEUTIC MODALITY FOR
TREATING TETANUS, NATURE OF
ANTI-TOXIN, THEY ARE PRESENT IN
THE ORGANISM, IMMUNE BODIES SO
CONSTITUTED THAT THEY ARE
ENDOWED NOT ONLY WITH AFFINITIES
OF AFFINITIES ...
THE ILLUSTRATION SHOWN HERE, SO
ANTIGEN IS CLEARLY DENOTED.
NEXT CLICK PLEASE.
IMMUNE BODY HAS A REGION OF
AFFINITY WHICH IS THE FAB
REGION.
CLICK AGAIN PLEASE.
FAB REGION.
NEXT REGION WOULD BE THE FC
REGION.
ALL OF THIS ALLOWS FOR
ENGAGEMENT IN FC RECEPTOR, SO
WHAT WE'VE COME TO RECOGNIZE IS
THAT THE IDEA BEHIND ANTIBODY
COUPLING TO EFFECTOR RESPONSES
IS NOT NEW.
WE'VE WORKED OUT DETAILS AND
JUST ILLUSTRATES THE IMPORTANCE
OF THE LEGACY OF THESE
REMARKABLE SCIENTISTS.
PHIL'S LEGACY LIVES ON IN EACH
OF US, IN THE WORK WE DO, THE
STUDENT WE TRAIN, STANDARDS WE
STRIVE FOR AND GOALS WE SET.
I COULDN'T BE MORE GRATEFUL TO
HIM AND HIS FIERCE DEVOTION TO
SCIENCE AT THE HIGHEST LEVEL.
THANKS, PHIL.
THANK YOU.
[APPLAUSE]
>> ONE MORE TALK BEFORE THE
COFFEE BREAK.
THAT'S ILAN KIRSCH, PEDIATRIC
RESIDENCY AT CHILDREN'S HOSPITAL
MEDICAL CENTER IN BOSTON,
FELLOWSHIP IN PEDIATRIC
HEMATOLOGY AND ONCOLOGY AT THE
NCI.
HE THEN DID A POSTDOC WITH PHIL
LEDER FROM 1979 TO 1982,
MOLECULAR GENETICS, YOU'VE SEEN
HIS PICTURE TEN TIMES ALREADY,
IN THE LABORATORY OF PHIL AT
NICHD AND SPENT 20 YEARS AT NCI
AS BASIC RESEARCHER, ATTENDING
PHYSICIAN IN PEDIATRIC BRANCH
ULTIMATELY CHIEF OF GENETICS
BRANCH, FOCUSED ON STRUCTURE AND
FUNCTION OF T-CELL RECEPTOR
GENES AND USE OF PROBES FROM
THESE LOCI TO STUDY
CANCER-SPECIFIC GENETIC
INSTABILITY, IN 2005 JOINED
AMGEN, HEADING ONCOLOGY RESEARCH
GROUP IN SEATTLE, HAS BEEN A
CONSULTANT WITH EXTENDED
ASSIGNMENTS FOCUSED ON
DEVELOPMENT OF ANTIBODY
THERAPEUTICS AND MOLECULAR
DIAGNOSTICS.
>> I WANT TO UNDERSCORE THE
SENTIMENT HEARD, WHAT AN HONOR
AND PLEASURE IT IS TO BE INVITED
TO JOIN THESE COLLEAGUES AND
SPEAK AT THIS DEDICATION OF THE
FAES PHIL LEDER RESIDENCE HOUSE.
WHEN I WAS A POSTDOCTORAL FELLOW
I KNEW I WAS IN A VERY GOOD LAB.
BUT I ASSUMED THAT IT WAS JUST
ONE OF MANY VERY GOOD LABS.
IT WAS ONLY AFTER LEAVING THE
LAB AND SETTING UP A RESEARCH
GROUP OF MY OWN AND VISITING AND
MORE CLOSELY OBSERVING THE LABS
OF OTHERS BOTH ACADEMIC AND
COMMERCIAL, THAT I REALIZED HOW
SPECIAL WAS THE LEDER LAB.
IT'S BEEN REINFORCED TODAY AS
I'VE LISTENED TO THE SPEAKERS
WHO HAVE PRECEDED ME SO FAR AT
THIS PODIUM.
AS A MEDICAL STUDENT, AND
RESIDENT I CAME TO KNOW STUART
ORKIN, THE FIRST TO SHOW ME HOW
TO DO A SOUTHERN BLOT.
WAS ACCEPTED INTO PHIL'S LAB,
BEFORE I LEFT BOSTON FOR
BETHESDA, I WAS SPEAKING WITH
STUART AND VOICING MY CONCERN
OVER WHETHER I'D BE ABLE TO
MEASURE UP TO THE CALIBER OF THE
PEOPLE IN THE LEDER LAB.
MANY YOU JUST HEARD SPEAK.
I REMEMBER DISTINCTLY THE
COMFORTING INSULT THAT STUART
GAVE ME, SAYING, DON'T WORRY,
LANNY, PHIL COULD GET GOOD
RESEARCH OUT OF A STONE.
OVER THE PAST 60 YEARS
ACHIEVEMENTS HAVE BEEN MENTIONED
BY OTHER SPEAKERS.
MY OWN LIST IS INCOMPLETE BUT
STILL DESERVES LISTING IN
REPUTATION.
ANY ONE OF THESE ACCOMPLISHMENTS
WOULD BE REASON ENOUGH TO BRING
US HERE TODAY TO HONOR PHIL
LEDER.
THE FACT HE WAS ABLE TO ACHIEVE
THEM ALL IS SIMPLY AWE
INSPIRING.
AND ALL THE WHILE THAT HE WAS
DOING THIS SCIENCE HE WAS A
PROTOTYPE, MENTOR, AND AS FAES
KNOWS WELL AN INCREDIBLY SKILLED
EDUCATOR.
WHEN A PERSON FROM HIS LAB WAS
SCHEDULED TO GIVE A TALK, HE
WOULD INVITE THAT PERSON INTO
THE BUILDING 6 6 LIBRARY AROUND
HAVE HIM OR HER DELIVER THE TALK
WITH SLIDES TO AN AUDIENCE OF
ONE, PHIL.
HE WOULD THEN QUESTION,
INSTRUCT, MODIFY, AND SUGGEST
WAYS THAT THE PRESENTATION MIGHT
BE IMPROVED.
WHAT A PRIVILEGE TO BE ABLE TO
HAVE THIS ONE-ON-ONE INSTRUCTION
BY A MASTER IN THE ART OF
SCIENTIFIC PRESENTATION AND
PUBLIC SPEAKING.
I, LIKE SO MANY PEOPLE, SPEAKING
TODAY, WAS DIRECTLY INTRODUCED
TO PHIL DURING ONE OF HIS PUBLIC
PRESENTATIONS.
MY FIRST EXPOSURE WAS AT THE
STILL AMAZING IN RETROSPECT COLD
SPRING HARBOR SYMPOSIUM ON
QUANTITATIVE BIOLOGY HELD IN
1976 ON LYMPHOCYTE DIVERSITY.
I REALLY CAN'T RECALL HOW I, A
MERE STUDENT, MANAGED TO GET
MYSELF INTO THAT MEETING.
PHIL SPOKE ON THE FINAL DAY,
INTRODUCING THE FOUNDATIONAL
PRINCIPLE OF VDJ RECOMBINATION,
PHIL'S TALK WAS A TOUR DU FORCE
PRESENTING WORK FROM HIS LAB.
THE AUDIENCE RECOGNIZED THE
IMPORTANCE OF WHAT IT WAS
HEARING.
I REMEMBER THAT JIM WATSON
SPECIFICALLY ASKED THAT THE TIME
CONSTRAINTS BE LIFTED SO THAT HE
COULD ASK PHIL AN ADDITIONAL
QUESTION.
A FEW YEARS AFTER THAT I WAS IN
PHIL'S LAB, AND A FEW YEARS
AFTER THAT PHIL LEFT NIH FOR
HARVARD.
ABOUT A YEAR OR SO AFTER PHIL
HAD LEFT THE NIH, HE WAS ASKED
TO RETURN TO GIVE A SPECIAL
NAMED LECTURESHIP, RIGHT HERE IN
THIS MASUR AUDITORIUM.
I WAS SITTING ABOUT TEN ROWS UP,
IN THE MIDDLE SECTION.
SO AS NOT TO MISS A SINGLE WORD
OR BIT OF DATA.
PHIL SHOWED A SLIDE ON THE
SCREEN.
A PICTURE OF A MOUSE.
PERHAPS IT WAS THIS VERY SAME
MOUSE.
FOR AT LEAST FIVE FULL MINUTES,
THAT MOUSE REMAINED THE ONLY
THING ON THE SCREEN WHILE PHIL
DETAILED THE STORY AND
IMPLICATIONS OF THE FIRST
ONCOMOUSE.
DURING THOSE MINUTES, AS THE
AUDIENCE SAT IN RAPT ATTENTION,
I FOUND MYSELF BOTH LISTENING
INTENTLY BUT ALSO TRYING TO
GLEAN ADDITIONAL PRACTICAL
KNOWLEDGE IN TERMS OF PUBLIC
SPEAKING.
HOW IS HE MANAGING TO DO THIS, I
ASKED MYSELF.
AS I MARVELED AT THIS EXAMPLE OF
ORATORY, I WAS INTERRUPTED BY A
STAGE WHISPER OF A PERSON BEHIND
ME, FOUR MINUTES IN TURNED TO
HIS FRIEND NEXT TO HIM AND SAID,
THIS GUY CAN DO ANYTHING!
HE COULD BE PRESIDENT!
[LAUGHTER]
YEP, EXACTLY, I THOUGHT TO
MYSELF.
I THINK THAT THE BROAD RANGE OF
PHIL'S SUCCESSFUL CONTRIBUTIONS
MAY HAVE AFFECTED ME TOO MUCH.
I WANTED TO EMULATE HIS REACH
AND GRASP, BUT REALIZED SOON
ENOUGH THAT THERE WAS ONLY ONE
PHIL LEDER.
I WORRIED MY OWN RESEARCH
ACTIVITIES WERE TOO DIFFUSE, I
WAS ENVIOUS OF COLLEAGUES
BUILDING SCIENTIFIC UNIVERSES
FROM THE GROUND UP.
LIKE STAN KORSMEYER, DEVOTING
HIMSELF TO THE PROGRAMMED CELL
DEATH PATHWAY OR JEFF RAPHETCH
WITH FC RECEPTORS.
A THEME GOVERNED MY CAREER
SPRINGING FROM MY TIME IN THE
LEDER LAB.
I'VE MADE A LIST OF INVOLVEMENTS
BUILT ON FOUNDATION OF GENOMIC
REARRANGEMENTS OF
IMMUNOGLOBULIN, AND T-CELL
RECEPTOR LOCI.
I THINK IF PHIL WERE LOOKING, HE
MIGHT THINK I HAD LEFT OUT A
PARTICULARLY IMPORTANT
INVOLVEMENT.
NAMELY, MY INTRODUCING HIM TO
THE MAGNIFICENT CYNTHIA MORTON.
POINT WELL TAKEN.
I'M GOING TO FOCUS THE NEXT PART
OF THIS TALK ON TWO PARTICULAR
SCIENTIFIC AND MEDICAL
APPLICATIONS OF THE WORK THAT
FIRST BEGAN FOR ME IN PHIL'S
LAB.
OF COURSE, IT BEGINS WITH THE
FOUNDATIONAL DIAGRAM THAT YOU
SAW MANY TIMES THAT SUMMARIZE
RESOLUTION OF THE QUESTION OF
THE ORIGIN OF LYMPHOCYTE
DIVERSITY, EXISTING IN OUR
GERMLINE DNA AS DISCRETE
DISCONTIGUOUS SEGMENTS, VARIABLE
FOR SOME LOCI.
THERE IS RANDOM ASSORTMENT THAT
PREVIOUSLY RANDOM ASSORTED WITH
Js TO FORM A NOW CONTIGUOUS
VDJ REGION.
THE SEQUENCE CREATED BY THESE
REARRANGEMENTS FORMS
HYPERVARIABLE REGION 3 OR CDR 3
PART OF IMMUNOGLOBULIN OR T-CELL
RECEPTOR.
THE PART OF THE MOLECULE MOST
RESPONSIBLE FOR SPECIFICITY.
THIS RECONFIGURATION OF GENOMIC
DNA CREATES A SPECIFIC MARKER OF
EACH LYMPHOCYTE THAT HAS REACHED
THE DEVELOPMENTAL STAGE WHERE
THIS RANDOM ASSORTMENT AND
RE-LIGATION OCCURS, THIS WAS
RECOGNIZED IN AND ELUCIDATED BY
PHIL IN THE LABORATORY BUT IN
ORDER FOR IT TO ACHIEVE ITS
CURRENT LEVEL OF PRACTICAL
APPLICATION, THREE BROAD STREAMS
OF KNOWLEDGE NEEDED TO BE
BROUGHT TOGETHER.
THE ORIGIN OF LYMPHOCYTE
DIVERSITY THAT I'VE JUST
DESCRIBED, MARKED IMPROVEMENTS
IN THE COST AND EFFICIENCY OF
DNA SEQUENCING, AND THE
COMPUTATIONAL ABILITY TO HANDLE
ENORMOUS AMOUNTS OF DATA.
THIS HAS RESULTED IN A PRACTICAL
SCALABLE AND ENORMOUSLY
INFORMATION RICH
IMMUNOSEQUENCING PROCESS, SHOWN
HERE.
IN WHICH FOUR PRIMERS RECOGNIZE
EACH AND EVERY KNOWN V SEGMENT
AND REVERSE PRIMERS RECOGNIZE
EVERY J SEGMENT FOR THE SAME
LOCUS, APPLIED TO DNA EXTRACTED
FROM ANY SAMPLE OF INTEREST, AND
AMPLIFICATION OCCURS ACROSS THAT
HIGHLY SPECIFIC CDR 3 REGION FOR
EACH AND EVERY B OR T CELL
PRESENT IN THE SAMPLE.
ALL AMPLIFIED MOLECULES ARE THEN
SEQUENCED.
THE NUMBER OF UNIQUE CDR 3s
CORRELATES WITH NUMBER OF UNIQUE
B OR T CELLS PRESENT IN THE
SAMPLE.
THE NUMBER OF TIMES ANY GIVEN
CDR 3 SEQUENCE IS SEEN
CORRELATES WITH THE SIZE OF THAT
B OR T CELL CLONE WITHIN THAT
SAMPLE.
EVERY TIME THE ASSAY IS RUN,
MORE INFORMATION IS GENERATED
THAN WAS GENERATED FOR THE FIRST
HUMAN GENOME SEQUENCE PROJECT.
AND THAT INFORMATION IS NOT
STATIC.
IT CHANGES OVER TIME, DEPENDING
ON WHAT ANTIGENS OR PATHOGENS AN
INDIVIDUAL ENCOUNTERS, OR FOR
THAT MATTER IF A PERSON DEVELOPS
A LYMPHOID MALIGNANCY.
SO, FOR EXAMPLE, IF APPLIED TO
BONE MARROW OF A HEALTHY
INDIVIDUAL, THE ENTIRE IMMUNE
REPERTOIRE OF THAT SAMPEL IS
PROFILED BUT NO ONE SEQUENCE OR
SEQUENCES STANDS OUT HIGH ABOVE
THE FREQUENCIES OF THE BULK
REPERTOIRE.
IN CONTRAST, IN A PATIENT WITH
LEUKEMIA LYMPHOMA OR MYELOMA,
BECAUSE OF THE DYSREGULATED
GROWTH OF THAT PARTICULAR CLONE,
THE CDR 3 SEQUENCE OR SEQUENCES
THAT RESIDE WITHIN THAT CLONE
ARE HIGHLY OVERREPRESENTED.
THAT SEQUENCE CAN THEN BE TAGGED
FOR SUBSEQUENT TRACKING DURING
THAT PATIENT'S COURSE OF
THERAPY, PROVIDING A DIRECT
DETERMINATION OF MEASURABLE
RESIDUAL DISEASE, MRD, IN THAT
PATIENT.
THIS IS THE BASIS FOR WHAT IS
NOW THE ONE AND ONLY FDA CLEARED
ASSAY IN PATIENTS WITH LYMPHOID
LEUKEMIA AND MYELOMA.
AT DIAGNOSIS, A HIGH TUMOR
BURDEN SAMPLE WAS RECEIVED,
REPORT LIKE THIS ONE IS
GENERATED.
THE NUMBER OF DOMINANT IMMUNE
RECEPTORS SEQUENCES RESIDING
WITHIN THE CLONE OF INTEREST ARE
ENUMERATED.
EACH DOMINANT SEQUENCE IS
PRESENTED ALONG WITH FREQUENCY
OF THAT SEQUENCE FOR TOTAL
NUCLEATED CELLS PRESENT IN THE
SAMPLE.
SAMPLE CLONALITY, NUMBER OF
UNIQUE CDR 3s, TOTAL NUMBER OF
CDRs 3 WHICH PROVIDES ESTIMATE
MUCH TOTAL NUMBER OF B OR T
CELLS IN THE SAMPLE ARE ALL
PROVIDED.
IF SOMATIC HYPERMUTATION IS
PRESENT IT IS NOTED, AND THE
SPECIFIC V SEGMENT INVOLVED IS
DEFINED.
IN SUBSEQUENT SAMPLES, THE
ENTIRE REPERTOIRE AGAIN IS
PROFILED, BUT THE SPECIFIC
SEQUENCE OR SEQUENCES IDENTIFIED
AS DOMINANT AT DIAGNOSIS ARE
SEARCHED FOR AND REPORTED, AS AN
INTEGER VALUE PER ONE MILLION
NUCLEATED CELLS, CLINICAL
REPORTS SENT TO ANY HEALTH CARE
PROVIDER WHO ORDERS THESE
CLONALITY AND TRACKING TESTS.
NOW HAVING ASSISTED IN CLINICAL
MANAGEMENT OF OVER 50,000
INDIVIDUAL PATIENTS, AND
HUNDREDS OF CLINICAL TRIALS.
INCREDIBLY INFORMATION RICH AND
FILLED WITH ADDITIONAL POSSIBLE
MEDICAL AND BIOLOGICAL INSIGHTS
OF WHICH WE'RE JUST BECOMING
AWARE.
THE SAME FUNDAMENTAL ASSAY CAN
BE EXTENDED TO ANY SITUATION IN
WHICH THE IMMUNE RESPONSE PLAYS
A ROLE, FOR LEUKEMIA LYMPHOMA
AND MYELOMA THE SEQUENCES FIND
US.
FOR OTHER SITUATIONS WE FIND THE
SEQUENCES THROUGH TWO
APPROACHES, ONE OF WHICH IS THE
REALIZATION OF ASPIRATION GOAL
SINCE VDJ RECOMBINATION WAS
REFINED, LINKAGE OF A SPECIFIC
T-CELL RECEPTOR NUCLEOTIDE
SEQUENCED TO THE TARGET OF THE T
CELL WITHIN WHICH THAT RECEPTOR
IS ENCODED.
THE METHODOLOGY OF OF THE
APPROACH IS STRAIGHTFORWARD.
T CELLS ARE ALIQUOTED AND
EXPOSED TO POOLS OF ANTIGEN
THROUGH PEPTIDE PULSING OR
TRANSFECTION, AND ENDOGENOUS
PRESENTATION BY
ANTIGEN-PRESENTING CELLS.
EACH ANTIGEN HAS A DISTINCT SET
OF WELL ADDRESSES ON THE
EXPERIMENTAL PLATE.
AFTER INCUBATION, CELLS FROM
EACH WELL ARE SORTED FOR
ACTIVATION MARKERS, AND THEN
SEQUENCED.
AND ACTIVATION MARKER POSITIVE
REPERTOIRES ARE DELINEATED
THROUGH DECONVOLUTION PROCESS,
RESPONDING TO, FOR EXAMPLE,
ANTIGEN 1 CAN BE DISTINGUISHED
FROM THOSE RESPONDING TO
ANTIGENS 2 OR 3, AND ALL OF
THOSE DISTINGUISHED FROM ANY
BACKGROUND NON-SPECIFIC
RESPONDING CELLS.
HUNDREDS OF T-CELL RECEPTORS
RESPONDING TO THOUSANDS OF
EPITOPES FROM VIRUSES,
AUTOIMMUNE ANTIGENS,
TUMOR-SPECIFIC ANTIGENS HAVE NOW
BEEN IDENTIFIED.
FOR EXAMPLE, EARLY ON IN THE
DEVELOPMENT AN ANALYSIS WAS
DONE AND PERFORMED AND PUBLISHED
BY THIS GROUP.
RESULTS OF T-CELL RECEPTOR
IDENTIFICATION OF THESE EPITOPES
FROM ONE ROUND OF MIRA FROM A
SINGLE PATIENT IS SHOWN HERE.
THE TUMOR ASSOCIATED GENES ARE
SHOWN IN DIFFERENT COLORS ON THE
X-AXIS WITH DIFFERENT EPITOPES
IN SEPARATE COLUMNS, EACH POINT
REPRESENTS A T-CELL RECEPTOR
ENRICHED BY EXPOSURE TO A
SPECIFIC EPITOPE.
AS THE EXPERIMENT IS REPEATED,
ADDITIONAL DONORS ARE ADDED AND
DONORS OF DIFFERENT HLA TYPES
ARE INCLUDED AND PATTERN OF
PUBLIC AND PRIVATE T-CELL
RESPONSES BECOMES MORE AND MORE
ROBUST.
THIS PROVIDES A MUCH MORE
ACCURATE AND SENSITIVE APPROACH
THAN ANY OTHER METHOD OF LINKAGE
OF T CELL TO TARGET.
ADDING TO THIS CAPABILITY IS THE
WELL KNOWN DESIGN OF CASE
CONTROL STUDIES.
IF WE CAN DEFINE A POPULATION
WITH A PARTICULAR EXPOSURE, FOR
EXAMPLE A RECENT POSITIVE PCR
TEST TO SARS-COV-2, AND HAVE A
CONTROL POPULATION FOR WHICH
WE'RE QUITE CERTAIN THERE WAS NO
EXPOSURE, FOR EXAMPLE SAMPLES IN
THE DATABASE PRIOR TO 2019, THEN
WE CAN INTERROGATE T CELL
REPERTOIRES FROM THE TWO
POPULATIONS AND EXTRACT A
CLASSIFIER SET OF T-CELL
RECEPTORS THAT IDENTIFY THOSE
WHO HAVE BEEN EXPOSED.
THE BULK OF THE RESPONSE FOR ANY
INDIVIDUAL WILL BE PRIVATE.
OCCURRING ONLY WITHIN THAT
INDIVIDUAL.
HOWEVER, THERE ARE ENOUGH SHARED
RECEPTORS AND MORE WITH
INCLUSION OF DIFFERENT HLA
TYPES, THIS WAS THE BASIC FOR
DIAGNOSTICS THAT RECEIVED
EMERGENCY USE AUTHORITIATION
AUTHORIZATION EARLY IN THE
PANDEMIC, IT TOOK LESS THAN TEN
MONTHS.
WHEN THE MIRA ASSAY IS COMBINED,
EVEN GREATER POWER IS ACHIEVED.
WE CAN DISSECT A DIFFERENTIAL
IMMUNOGENICITY OF DELTA OR
OMICRON COVID VARIANTS AND KNOW
PRECISELY WHICH T CELLS WILL
STILL RECOGNIZE A NEW EMERGENT
VIRUS, WE CAN DISTINGUISH T
CELLS THAT RECOGNIZE THE SPIKE
FROM NON-SPIKE PARTS OF THE
VIRAL GENOME, AND IN SO DOING
DISTINGUISH VACCINE RESPONSE
FROM RESPONSES DUE TO ENDOGENOUS
INFECTION.
THESE EXAMPLES THAT I PROVIDED
ARE JUST A SNAPSHOT OF THE POWER
OF IMMUNOSEQUENCING, THE ABILITY
TO ENUMERATE, SPECIFY, QUANTIFY
EACH AND EVERY B AND/OR T CELL
IN ANY SAMPLE OF INTEREST.
IT REINFORCES THE FUNDAMENTAL
REALIZATION THE IMMUNE SYSTEM IS
AN EVOLUTIONARY MARVEL AND
IMMUNOSEQUENCING OFFERS A
SPECIFIC STANDARDIZED AND
SENSITIVE METHOD FOR
DETERMINATION, PROVIDES
CLASSIFIERS FOR EVALUATION OF
IMMUNE RESPONSIVENESS TO
SPECIFIC ANTIGENS AND PATHOGENS,
HIGH-THROUGHPUT AND SCALABLE
PROCESS IS NOW AVAILABLE FOR
LINKING A T-CELL RECEPTOR
NUCLEOTIDE SEQUENCED TO TARGET
OF THE RECEPTOR THAT SEQUENCE
ENCODES, IMMUNOSEQUENCING LEADS
TO SPECIFIC IMMUNOGLOBULIN AND T
CELL THERAPEUTICS.
IMMUNE REPERTOIRE IS A DIARY,
IMMUNE RECEPTOR PROFILING CAN
PROVIDE INSIGHT INTO ONCOLOGY,
INFECTIOUS DISEASE,
AUTOIMMUNITY, AGING, ET CETERA.
I WANT TO CONCLUDE BY RETURNING
TO WHERE MY PERSONAL STORY WITH
PHIL BEGAN.
YOU'VE SEEN THIS PICTURE MANY
TIMES.
THIS IS A PICTURE OF PHIL FROM
AROUND THE TIME OF THAT COLD
SPRING HARBOR SYMPOSIUM I
MENTIONED EARLIER.
IN CASE IT'S NOT APPARENT SO
FAR, NOTE THE DIAGRAM ON THE
BOARD BEHIND HIM.
I WISH THAT PHIL COULD BE HERE
TO BE ACKNOWLEDGED FOR THE
PROGRESS THAT HAS BEEN MADE IN
THE APPLICATION OF THE
FOUNDATIONAL PRINCIPLES OF VDJ
RECOMBINATION TO BIOLOGY AND
MEDICINE.
I'M SURE HE WOULD REFLECT THAT
WE'VE ONLY SCRATCHED THE SURFACE
OF THE IMPLICATIONS AND
APPLICATIONS OF THIS PROCESS.
I WISH THAT IN PREPARATION FOR
THIS TALK, I MIGHT HAVE ONCE
AGAIN BEEN AT THE PODIUM IN THE
LIBRARY OF BUILDING 6 WITH PHIL
SEATED BY HIMSELF NO MORE THAN
TEN FEET AWAY, STOPPING ME TO
SUGGEST A BETTER SLIDE TO SHOW
OR PHRASE TO USE.
FOR YOU ALL, IT WOULD HAVE MADE
THE PAST 20 MINUTES MORE
ENJOYABLE AND INSTRUCTIVE.
IT WOULD ALSO HAVE PROVIDED ME
WITH ADDITIONAL INSIGHTS AND
UNDERSTANDING OF MY OWN WORK AND
THE WORK OF OTHERS, INSIGHTS
THAT FRUSTRATINGLY I MAY NOW
NEVER REALIZE.
I WANT, AGAIN, TO THANK THE FAES
FOR ITS RECOGNITION AND TRIBUTE
TO PHIL IN THE BUILDING THAT NOW
IS DEDICATED TO HIM, AND THANK
YOU ALL FOR ALLOWING ME TO SHARE
MY RECOLLECTIONS AND
PERSPECTIVE.
[APPLAUSE]
>> THANK YOU.
WE'LL TAKE A TEN-MINUTE BREAK.
PLEASE ONLY TAKE TEN MINUTES AND
BE BACK AT 11:41.
THANK YOU.
OUR NEXT SPEAKER IS KATHLEEN
KELLY, WHO IS A DEPUTY
SCIENTIFIC DIRECTOR, CENTER FOR
CANCER RESEARCH, AND CHIEF OF
THE LABORATORY OF , RECEIVED A
Ph.D. IN CELLULAR IMMUNOLOGY
FROM UNIVERSITY OF CALIFORNIA
IRVINE, JOINED THE LEDER LAB AT
HARVARD MEDICAL SCHOOL, WHILE IN
THE LAB KATHY MET AND MARRIED
ANOTHER POSTDOCTORAL FELLOW IN
THE LEDER LAB, AFTER COMPLETING
THEIR POSTDOCTORAL TRAINING BOTH
ESTABLISHED PROGRAMS AT NIH AND
NIAID AT NIH, DR. STEPHEN LIST
WHO STUDIED THE NF-kappaB
FAMILY OF TRANSCRIPTION FACTORS
PASSED AWAY FROM PANCREATIC
CANCER IN 2020, WE WISH HE
COULD HAVE BEEN WITH US BUT
KATHY PROMISED TO TALK ABOUT HIS
CONTRIBUTION AS WELL.
SHE FOCUSES ON ONCOGENESIS,
ADVANCED PROSTATE CANCER AS
RELATED TO ASSOCIATED DRUG
VULNERABILITIES AND RESISTANCE.
>> THANK YOU VERY MUCH.
IT REALLY IS A DISTINCT PLEASURE
TO BE HERE TO HONOR PHIL LEDER,
AND WHAT I WILL DO IS SHARE SOME
MEMORIES WITH YOU.
LIKE THOSE THAT YOU'VE ALREADY
HEARD THIS MORNING, THEY WILL
REALLY UNDERSCORE NOT ONLY
PHIL'S IMMENSE SCIENTIFIC
TALENT, ALSO HIS DEDICATION TO
MENTORS AND SUPPORTING THE
SCIENTIFIC COMMUNITY.
HIS LEGACY AT NIH IS HUGE,
COULDN'T BE MORE APPROPRIATE TO
HAVE A PHIL LEDER HOUSE TO
PROMOTE SCIENTIFIC TRAINING.
I'M ONE OF THE SMALL GROUP OF
TRAINEES WHO BRIDGE NIH AND
HARVARD YEARS.
I THINK I MAN THE ONLY ONE IN
THIS GROUP WHO IS HERE TODAY.
I COMPLETED MY GRADUATE TRAINING
AT U.C. IRVINE IN CELLULAR
IMMUNOLOGY, AND DURING MY
GRADUATE YEARS IMMUNOGLOBULIN
REARRANGEMENTS WERE DISCOVERED
THAT YOU'VE BEEN HEARD ABOUT,
AND THIS WAS VERY CONCEPTUALLY
TRANSFORMATIVE STIMULATING OUR
IMAGINATIONS ABOUT UNKNOWN
POSSIBILITIES OF GENOMIC
STRUCTURE AND FUNCTION TO COME.
IT ALSO UNDERSCORED THE VALUE
AND IMPORTANCE OF OBTAINING
MOLECULAR GENETIC SKILLS AS PHIL
HIETER POINTED OUT.
I COULD HARDLY BELIEVE MY GOOD
FORTUNE WHEN PHIL INVITE
IMMEDIATE TO JOIN HIS LAB.
I ARRIVED IN SUMMER OF 1980,
PHIL JUST ACCEPTED A POSITION AT
CHAIR OF GENETICS AT HARVARD,
ASKED ME TO MOVE WITH THE LAB
THE FOLLOWING SPRING, THIS WAS
FINE WITH ME.
I HAD SPENT MY LIFE TO THAT
POINT ON THE WEST COAST, AND I
WAS HAPPY TO EXPLORE MORE OF THE
EAST COAST.
MY FIRST PROJECT YOU'VE SEEN
PICTURES OF WHERE WE WERE IN
BUILDING 6, WAS AN AMBITIOUS
ONE.
TO CLONE THE T-CELL RECEPTOR.
ULTIMATELY, OUR APPROACH DIDN'T
PAN OUT BUT IN THE PROCESS I
LEARNED HOW TO DO MOLECULAR
GENETICS.
THIS FIRST YEAR TURNED OUT TO BE
AN IMMENSELY EXCITING TIME,
RECURRING CHROMOSOMAL
TRANSLOCATION IN BURKITT'S
LYMPHOMA AND CML WERE
IDENTIFIED, LANNY'S BRILLIANT
MOVE TO INTRODUCE CYNTHIA TO
PHIL, THE CONCEPT OF ONCOGENES,
CELLULAR HOMOLOGS OF GENES AND
TRANSFORMING RETROVIRUSES WAS
RAPIDLY EVOLVING, THE
TRANSFORMATION ASSAY HAD
IDENTIFIED THE RAS ONCOGENES,
ONCOGENE AND OTHERS.
AND SO AS YOU'VE HEARD FROM
LANNY, HE, BECKY AND CYNTHIA
IDENTIFIED MYC ONCOGENE IN
BURKITT'S LYMPHOMA.
FROM THE VANTAGE NOW OF HAVING
DIRECTED COMPETITIVE PROJECTS IN
MY OWN LAB, I CAN NOW REALLY
APPRECIATE HOW EXCITED PHIL WAS
BY SCIENCE BUT HOW HE MAINTAINED
A DIGNIFIED DEMEANOR.
THIS FIRST YEAR WASN'T ONLY
SCIENTIFICALLY MOMENTOUS BUT
PERSONALLY MOMENTOUS, AS MICHAEL
JUST SAID PRIOR TO MOVING TO
BOSTON I MARRIED ANOTHER POSTDOC
FELLOW IN PHIL'S LAB, AND I'LL
TALK ABOUT BOTH OF OUR CAREER
JOURNEYS.
SO, PRIOR TO JOINING PHIL'S LAB
ULI WAS A GRAD STUDENT AT
HARVARD DURING THE DEVELOPMENT
OF SEQUENCING, RNA POLYMERASE
INTERACTION WITH PROKARYOTIC
PROMOTERS.
HERE IS A PICTURE OF STEVE ON
THE LEAVE, ULLI AND CARL WU.
I CHOSE THIS PHOTO BECAUSE ALL
THREE OF THESE SCIENTISTS SPENT
AT LEAST PART OF THEIR
SCIENTIFIC CAREER HERE AT NIH.
SO, ULLI JOINED PHIL'S LAB, AND
DURING THE FIRST FEW YEARS
WORKED ON IMMUNOGLOBULIN GENE
STRUCTURE AND REARRANGEMENT
WHICH FOR THE IMMUNOLOGISTS IN
THE AUDIENCE INCLUDED DISCOVERY
OF D SEGMENTS.
A SMALL GROUP WENT WITH PHIL
FROM NIH TO HARVARD, HERE YOU
SEE ULLI, AND TIM STEWART JOINED
US IN HARVARD, I WANTED TO
INCLUDE THIS PICTURE.
TIM WAS THE PERSON THAT MADE THE
ONCOMOUSE THAT YOU'VE HEARD MUCH
ABOUT.
SO, ONE MEMBER OF THE GROUP THAT
SPENT TIME AT BOTH NIH AND
HARVARD WAS JIM BATTEY, WHICH
UNFORTUNATELY HE'S NOT HERED
THAT BUT JIM RETURNED TO NIH AND
EVENTUALLY BECAME DIRECTOR OF
NIDCD, PRIOR TO HIS RETIREMENT A
FEW YEARS AGO.
JUST ANOTHER EXAMPLE OF PHIL'S
NIH LEGACY.
ANYWAY, DURING THE THAT TIME AT
HARVARD THE FIELD OF ONCOGENES
EXPLODED, HOW DO THEY WORK?
NICK WAS PARTICULARLY PUZZLING
IN THE SENSE THERE WERE NO CLEAR
SOMATIC MUTATIONS.
SO, DRAWING ON MY BACKGROUND IN
CELLULAR IMMUNOLOGY WHERE I
WORKED ON B CELL ACTIVATION, AND
USING A NEWLY DEVELOPED AND
SENSITIVE RNAASE ASSAY AS YOU'VE
HEARD PHIL ALWAYS ENABLED NEW
TECHNOLOGIES AND PROMOTED THEM
IN THE LAB, I WAS ABLE TO
DETERMINE THAT MYC RNA WAS
TRANSCRIPTIONAL INDUCED BY B
CELL MITOGENS, IT MIGHT FUNCTION
AS A RESULT OF EXCESSIVE OR
MISTIMED EXPRESSION.
IN ADDITION, WE WORKED WITH
CHUCK STYLES AT DANA-FARBER TO
DEMONSTRATE THE SAME PATTERN OF
MITOGEN RESPONSIVNESS AND AT THE
TIME THE TOOLS WEREN'T ABLE TO
LOOK AT MORE THAN ONE GENE AT A
TIME, AND THIS OBSERVATION OF
MYC REGULATION WAS OF GREAT
INTEREST TO OUR ONCOGENE
COMMUNITY.
SO WE WROTE UP THE PAPER AND
DECIDED TO TRY A SUBMISSION TO
"CELL."
PHIL DID SOMETHING AT THIS POINT
THAT MEANT A LOT TO ME,
EXEMPLIFIES HOW HE SUPPORTED AND
PROMOTED HIS TRAINEES.
THE CELL OFFICES WERE IN
CAMBRIDGE.
WE PACKAGED UP OUR SUBMISSION
INTO MANILA ENVELOPE, I THINK WE
WROTE BEN'S NAME ON THE FRONT
WITH FELT WRITING PEN.
SO, THERE WAS NOTHING LIKE
ONLINE SUBMISSIONS IN THOSE
DAYS.
AND PHIL DROVE ME OVER THE RIVER
WHERE I HANDED IN OUR
SUBMISSION.
ON THE WAY HOME HE INSISTED WE
STOP AND EACH GET A GIANT COOKIE
FROM THE BAKERY TO CELEBRATE.
THE PAPER WAS ACCEPTED,
FOLLOWING THIS PUBLICATION PHIL
GENEROUSLY PASSED ON TO ME MANY
OF THE SEMINAR AND MEETING
INVITATIONS HE RECEIVED TO TALK
ABOUT THE WORK AND CRITICIZED MY
TALK AS LANNY HAS DESCRIBED TO
YOU.
SO, ULLU USED HIS EXPERIENCE IN
CHROMATIN STRUCTURE MAPPED THE
SEQUENCE.
SHORTLY THEREAFTER, IN EARLY
1984, ULLI AND I RETURNED TO NIH
TO SET UP AN INDEPENDENT LAB, AT
NIAID AND NCI.
WE WORKED TOGETHER TO ISOLATE
AND IDENTIFY GENES THAT WERE
TRANSCRIPTIONALLY REGULATED IN
HUMAN ACTIVATED T CELLS,
NF-kappaB BECAME THE FOCUS OF
ULLI'S WORK HE PASSED AWAY IN
2020, AS YOU HEARD.
WHEN PHIL WAS ON CAMPUS HE WOULD
SAY HI, CHECK UP ON THE FAMILY,
SHARE NEWS ABOUT HIS FAMILY.
HE ALWAYS STAYED IN TOUCH.
SO, I'LL RETURN TO MY OWN WORK
IN A MOMENT.
AND END WITH A VIGNETTE OF
INVESTIGATIONS TO GIVE YOU A
FLAVOR OF WHERE I HAVE ENDED UP
AFTER 40 YEARS.
BEFORE DOING THAT, I JUST WANT
TO SAY IT IS SO APPROPRIATE THAT
THE LEDER HOUSE WOULD BE HOME TO
STUDENTS WHO PARTICIPATE IN
COLLABORATIVE AND LARGELY
MULTI-DISCIPLINARY PROJECTS.
AS A SCIENTIST, PHIL HAD A RANGE
OF INTEREST IN BIOLOGICAL
MEDICINE, BIOLOGICAL MECHANISMS
AND MEDICINE, INCLUDING
ONCOLOGY, IMMUNOLOGY,
DEVELOPMENT, GENE REGULATION.
HE SET AN IMPORTANT EXAMPLE FOR
ALL OF US TO CONDUCT ETHICAL AND
COMPLETE RESEARCH WHILE
MAINTAINING RESPECT AND HUMANITY
TOWARD OTHERS.
SO, I'D LIKE TO SHARE AN E-MAIL
I RECEIVED, LET'S SEE.
THIS IS GOING TO BE HARD BECAUSE
I THINK THE E-MAIL GOT BLACKED
OUT SOMEHOW.
OKAY, THANK YOU.
THAT'S GREAT, OKAY.
WE'RE GOOD.
SO I WANT TO SHARE THIS E-MAIL
THAT I RECEIVED A FEW DAYS AGO
FROM HOWARD YOUNG, WHO LIKE ME
IS AN INVESTIGATOR FOR MANY
YEARS AT THE NCI.
SO, HE WROTE, I WANTED TO
MENTION ONE INTERACTION I HAD
WITH DR. LEDER THAT I NEVER HAVE
FORGOTTEN, WORKING AT THE LABS
WHEN THE COMPANY ANNOUNCED
ONE-THIRD OF EMPLOYEES WERE
BEING TERMINATED.
I WAS NOT IN THE GROUP.
DR. LEDER TOLD ME HE WOULD HELP
ME FIND A NEW POSITION.
THIS WAS INCREDIBLY GRACIOUS OF
HIM, ESPECIALLY SINCE WE DID NOT
HAVE A LOT OF PERSONAL
INTERACTIONS.
TO THIS DAY, I'M NOT SURE WHAT
PROMPTED HIM TO REACH OUT TO ME
BUT IT CERTAINLY HAS REMAINED
WITH ME ALL THESE YEARS.
SO, THIS MADE ME THINK ABOUT THE
FACT THAT THERE IS LIKELY SO
MUCH PHIL HAS DONE FOR OTHERS
THAT WE CANNOT FULLY APPRECIATE
EVEN TODAY.
SO, FINALLY THEN I'LL TURN TO MY
OWN WORK, WHICH HAS REVOLVED
AROUND ONCOGENIC MECHANISMS FOR
THE MOST PART, IN THE PAST 20
YEARS I'VE CONCENTRATED ON
ADVANCED PROSTATE CANCER, PHIL
WAS AN INNOVATOR PUSHING BOUND
A RIES.
IN INVESTIGATING PROSTATE
CANCER, ONE OF THE MOST
PERSISTENT PROBLEMS HAS BEEN A
LACK OF MODELS THAT RECAPITULATE
RANGE OF HUMAN DISEASE.
IN RESPONSE TO THAT CHALLENGE,
MY LAB HAS DEVELOPED A PLATFORM
OF MODELS THAT PREVIOUSLY
COULDN'T BE MAINTAINED IN A
CULTURE DISH TO WORK WITH.
BUT USING A NEW TYPE OF
CULTURING METHOD, CALLED
ORGANOID CULTURES, WE HAVE BEEN
ABLE TO ESTABLISH A MORE
REPRESENTATIVE APPROACH.
SO, CAN WE GO BACK TO THE WHITE
BACKGROUND MAYBE AND THE BLACK
LETTERING?
WOULD THAT BE POSSIBLE?
SOMETHING CHANGED SOMEWHERE.
WELL, I'LL GET STARTED, IF YOU
CAN PUT IT BACK TO THE ORIGINAL
WAY, THE SLIDE WAS, WITH THE
WHITE BACKGROUND AND BLACK
LETTERING THEN WE COULD INCLUDE
EVERYTHING.
SO, WHAT I'M SHOWING HERE IS A
SUMMARY OF THE SEQUENCE OF
EXPERIMENTS THAT SPANS SEVERAL
YEARS, AND THAT START BY
DETERMINING THERAPEUTIC RESPONSE
TO ABOUT 100 DRUGS, ACROSS --
GREAT, THANK YOU, ACROSS 35
MODELS OF ADVANCED PROSTATE
CANCER.
AND WHAT'S EMPHASIZED HERE IS
THE RESPONSE TO A COMMONLY
DOCETAXEL THAT INHIBITS FOR A
WHILE IN A PORTION OF ADVANCED
PROSTATE CANCER PATIENTS.
SO, HAVING A LARGE GROUP OF
RESPONDERS AND NON-RESPONDERS
ALLOWED US TO IDENTIFY A
CONSISTENT DIFFERENCE BETWEEN
GROUPS.
AND THAT DIFFERENCE IS THAT
NON-RESPONDERS EXPRESSED THE
HNF1 TRANSCRIPTION FACTOR.
SO, LOOKING MORE INTO THE
MECHANISM OF ACTION, WE
DETERMINED THAT THE PRESENCE OF
HNF1 STIMULATES EXPRESSION OF
ANOTHER PROTEIN, BIRK 3, THAT
HELPS TREATED CELLS.
THERE'S A CLASS THAT INHIBITS
THE ACTION OF BIRK 3, ACTIVE IN
MODEL THAT EXPRESS HNF1 AND DO
NOT RESPOND TO DOCETAXEL.
WHAT THIS MEANS, POSSIBLE TO
IDENTIFY PATIENTS THAT MAY NOT
RESPOND TO DOCETAXEL AND THERE
MAY BE A TREATMENT THAT WILL
COMPLEMENT DOCETAXEL FOR
NONRESPONSIVE PATIENTS.
I'M GOING TO END AND AGAIN JUST
SAY PHIL'S LEGACY AT NIH IS
SIMPLY IMMENSE, HE'S TOUCHED
MANY OF US, CERTAINLY MY LIFE.
THANK YOU VERY MUCH FOR YOUR
ATTENTION.
[APPLAUSE]
>> NEXT IS LOTHAR HENNIGHAUSEN,
GREW UP IN GERMANY, RECEIVED A
Ph.D. IN GENETICS FROM
UNIVERSITY OF C COLOGNE, JOINED
THE LEDER LAB IN 1983, RECRUITED
TO THE NIH, CHIEF OF THE
GENETICS AND PHYSIOLOGY SECTION
IN LABORATORY OF CELL AND
MOLECULAR BIOLOGY.
HIS RESEARCH FOCUSES ON HOW
CYTOKINES CONTROL ORGAN
PHYSIOLOGY, INCLUDING MAMMARY
GLAND DURING PREGNANCY.
WELCOME.
>> I BROUGHT THIS BOOK WHICH
PROBABLY IS THE MOST IMPORTANT
BOOK I EVER READ.
WE TALKED ABOUT TECHNOLOGY, THIS
AFFECTED ME DURING MY Ph.D.,
IN GERMANY TASKED WITH CLONING
GENES FROM THE MOUSE MAMMARY
GLAND.
YOU HAD TO ISOLATE MESSENGER
RNA, FIRST STEP TO ISOLATE
GENES.
AND THE FIRST PAPER I READ WAS
1972, CELLULOSE TO ISOLATE
MESSENGER RNA.
YOU COULDN'T JUST ORDER FROM THE
COMPANY, YOU MIXED IT TOGETHER,
LOWED COLUMNS.
YOU CAN SEE AN EXPERIMENT OF A
Ph.D. STUDENT PUBLISHED A FEW
YEARS LATER, ON THE LEFT INPUT
RNA, 28 AND 18 RNS, ON THE RIGHT
YOU CAN SEE ENRICHMENT, ARROWS
POINT TO ABUNDANT SPECIES CLONED
LATER ON.
1982, ONE SENTENCE TO POINT OUT
IS I THINK YOU WILL FIND THE
ATMOSPHERE IN OUR LABORATORY
BOTH FRIENDLY AND STIMULATING.
AND THIS WAS ABSOLUTELY TRUE.
WHAT WAS IT HERE?
THIS OFFER PRESUMES YOU'LL BE
ABLE TO OBTAIN FELLOWSHIP
SUPPORT, I SECURED A FELLOWSHIP.
YOU MAY RECOGNIZE SOME PEOPLE.
THIS MUST BE CYNTHIA HOLDING A
CAN OF BEER, PHIL AND I, TALKING
TO MYSELF, OH, AND CHRIS OVER
THERE.
WE TALKED ABOUT WHAT WE LEARNED
FROM PHIL, MESSAGES HE PASSED ON
TO US.
AND ACTUALLY ALMOST EVERYBODY
HAS THE FIRST ONE, IDEAS ARE
CHEAP.
WE ALWAYS BOUNCED IDEAS OFF HIM.
HE ALWAYS SAID IDEAS ARE CHEAP,
YOU HAVE TO FOLLOW UP.
STILL TRUE TODAY.
HE SAID, I'M A LINEAR THINKING,
IN THE MODERN WORLD HE WOULD SAY
I'M A STRATEGIC THINKER.
THIS ALSO I LEARNED FROM HIM.
LENNY AND KATHY MENTIONED
ALREADY THAT WE HAD TO GIVE
PRACTICE TALKS TO PHIL.
PHIL WAS INVITED TO GIVE A
LECTURE AT THE CONFERENCE AT THE
MIDWEST IMMUNOLOGISTS, DOES THIS
STILL EXIST?
THEY ARE BASED IN CHICAGO, NOT
SURE THE ORGANIZATION STILL
EXISTS.
PHIL DIDN'T WANT TO GO AND SENT
ME.
THEN I GAVE MY PRACTICE TALK TO
PHIL.
AND AT THE END OF THE TALK HE
SAID, LOTHAR, NOBODY IN THIS
AUDIENCE WILL HAVE FOLLOWED YOUR
WORK.
THEY WOULDN'T HAVE UNDERSTOOD A
SINGLE WORD YOU SAID.
WE WENT THROUGH THE SLIDES AND I
THINK I IMPROVED THE TALK TO
SOME EXTENT.
I'M STILL STRUGGLING WITH THAT.
ONE OF OUR LAST VISITS,
PRISCILLA, MY WIFE AND I,
VISITED A COUPLE YEARS AGO, PHIL
AND AYA AND I WENT OUT FOR
DINNER.
HE SAID IN MY CAREER, I GAVE IT
ALL.
THIS REFLECTS HIM, GIVING IT
ALL, THE PASSION.
SO NOW LET'S GO TO PHILS LAB,
1983, KATHY INTRODUCED THE LIST.
LOOKING FOR ELEMENTS THAT COULD
REGULATE GENES ESPECIALLY C-MYC,
ENHANCERS JUST BEING DISCOVERED
AT NIH, ULLI SET OUT TO MAP
CANDIDATE ENHANCERS USING DNA
HYPERSENSITIVITY STUDIES, OPEN
CHROMATIN WHICH WOULD NOT BIND
TO HISTONES BUT REGULATORY
PROTEINS.
AND MY TASK WAS ACTUALLY -- PHIL
WAS KIND, HE SAID JUST TALK TO
PEOPLE AND FIND A PROJECT.
HE DIDN'T IMPOSE ANY PROJECT.
JUST FIND ONE WHICH INTERESTS
YOU.
I WAS INTERESTED IN PROTEINS
BINDING TO DNA.
AND, AGAIN, HOW DO YOU IDENTIFY
PROTEINS BY DNA?
CHIP-seq, RNAseq, BACK IN
THE DAY IT WAS MORE PRIMITIVE.
ALL OF YOU KNOW THIS PICTURE.
IF YOU GO TO THE MUSEUM AT NIH
YOU'LL SEE PHIL'S ORIGINAL
DEVICE USED TO CRACK THE GENETIC
CODE WITH NUREMBERG, IT WAS
PATENTED.
I USE THIS DEVICE, A MORE
UPDATED VERSION, THE WAY WE
IDENTIFIED DNA BINDING PROTEINS
WE LAID DNA FRAGMENTS, KINASE,
FRAGMENTS INCUBATED WITH NUCLEAR
EXTRACTS.
AND THEN YOU PASSED OVER THE
MEMBRANE IN THE FILTRATION
APPARATUS.
FREE DNA, NOT BOUND TO DNA,
FLOWING THROUGH THE MEMBRANE,
NITRO CELLULOUS.
THEY WERE LABELED.
ON THE LEFT SIDE YOU CAN SEE THE
INPUT DNA WITHOUT PROTEIN,
FRAGMENTS WENT THROUGH THE
MEMBRANE, RIGHT-HAND SIDE,
NUCLEAR EXTRACT, YOU CAN SEE
PROTEIN BIND TO THE UPPER
FRAGMENT.
THIS WAS IN A PAPER IN 1984, THE
DRIVING FORCE IN THIS STUDY,
GENE REGULATION IN BURKITT
LYMPHOMA.
WE WANT TO KNOW WHERE PROTEINS
BIND, YOU USE YOUR FRAGMENTS,
DUMP THE PROTEIN, PARTIAL
DIGESTION.
IT'S A FOOTPRINT, RESISTANT TO
ONE DIGESTION.
AND WE DIDN'T KNOW WHAT THE
PROTEINS WERE AT THAT POINT, NOW
CALLED STAT PROTEINS AND NFIB
FOLLOWED MY CAREER FOR THE NEXT
ALMOST 40 YEARS.
SO, TIME TO MOVE ON, AND
EVERYBODY ELSE MOVED TO THEIR
OWN CAREER, GOT THE SOLUTION
BOOK, THE IDEA WAS YOU ACTUALLY
RECORD ALL THE SOLUTIONS YOU
GENERATE AND ALL THE CLONES AND
ALL THE IMPORTANT THINGS
INCLUDING PICTURES.
AND ALWAYS THE PERSON WHO LEFT
THE LAB WAS NUMBER ONE.
AND THIS WAS ME BACK IN THE DAY,
THOSE OF YOU WHO KNOW THE LAB IN
BOSTON THIS WAS THE DOOR TO
PHIL'S OFFICE.
ACTUALLY I SHARED A BENCH WITH
PHIL.
AND NORMALLY PHIL DIDN'T WORK
TOO MUCH AT THE BENCH BUT WHEN
ONCOMASS CAME ALONG HE WAS THERE
EVERY DAY, HIS WHITE LAB COAT,
CAMERA, AROUND ALL THE TIME.
HE SAYS, LOTHAR, IN 1983 IN HIS
TEMPORARY CORNER, I WAS SUPPOSED
TO GET A PROPER DESK BUT I STATE
FOR MORE THAN TWO YEARS ON THIS
TINY DESK WITH THE GEL BOX NEXT
TO IT.
VERY CONVENIENT.
SO I MOVED ON STARTING SOMETHING
NEW, FOLLOWING YOUR DREAMS,
STARTING SOMETHING NEW.
AND I'VE CLONED THE GENES FROM
THE MAMMARY GLAND IN THE MOUSE
AND DECIDED HAVING LEARNED NEW
THINGS IN PHIL'S LAB MOUSE
GENTLES COULD BE REGULATED, WENT
BACK TO THIS.
WHY IS THIS SUCH A FASCINATING
PROBLEM?
MAMMARY GLAND IS AN ORGAN ONE
SOLE PURPOSE TO PRODUCE LARGE
QUANTITIES OF MILK, IN THE MOUSE
THE MOUSE COULD DO PRODUCE 20%
OF BODY WEIGHT IN MILK EVERY
DAY.
YOU CAN SEE THE MEMORY ALVEOLI,
PURPOSE TO MAKE MILK PROTEINS,
ACCOUNTING FOR 90% OF MESSENGER
RNA IN THOSE CELLS, ONE REASON
WHY EASY, IN QUOTATION MARKS, TO
CLONE GENES.
NEXT DECADE WE USED TECHNOLOGIES
TRANSGENIC ANIMALS, KNOCKOUT
ANIMALS TO IDENTIFY HOW GENES
ARE REGULATED.
ONE KEY TRANSCRIPTION FACTOR IN
THE '90s, STAT5, KEY DRIVER OF
MAMMARY GLAND DEVELOPMENT.
IF YOU KNOCK OUT STAT5 THERE'S
NO MAMMARY GLAND DEVELOPMENT
DURING PREGNANCY MUCH IT'S
IMPORTANT FOR ACTIVATION OF
GENES, WE IDENTIFY USING MOUSE
GENETICS AND MUTATIONS AND
DELETIONS.
HOW THREE COMPONENTS DRIVE
10,000 FOLD, PROMOTERS,
ENHANCERS, SUPERENHANCERS,
DEPENDENT ON STAT5.
MOST GENES IN THE MAMMARY GLAND
ARE SINGLE GENE LOCI, SOME ARE
IN CLUSTERS.
AND THERE'S ONE LOCUS WHICH IS
QUITE UNIQUE, IT'S CALLED
CASEINS, IN THE MAMMARY GRANDS,
OTHERS IN THE SALIVARY GLAND,
OTHERS EXPRESSED IN MAMMARY
ANDEL IS -- SALIVARY
WHAT IS THE LOCATION OF SUCH A
COMPLEX LOCUS WITH EIGHT
DIFFERENT GENES?
ARE THERE ANY SPECIFIC ENHANCER
ELEMENTS?
LEE SOLVED THE QUESTION AND
IDENTIFIED SUPER ENHANCER, AND
WE THOUGHT THE SUPER ENHANCER
WOULD COVER THE ENTIRE LOCUS,
RESPONSIBLE FOR ACTIVATION OF
ALL THE GENES, BUT IN THE
MAMMARY GLAND THIS CONTROLS ONLY
TWO MAMMARY SPECIFIC GENES AND
ONE SHARED GENES.
THE OTHER PROMOTERS, OWN
ENHANCERS, RESTRICTED.
THIS LOCUS IS UNIQUE,
INTERSPERSED IN MAMMARY AND
SALIVARY GLAND, SUPER ENHANCER
IS DUAL ACTIVITY, ACTIVATE GENES
IN MAMMARY GLAND OR SPECIFICALLY
SALIVARY GLAND.
HIGHLY ACTIVATED GENES.
STAT5 IS CENTRAL, BUT THE NEXT
QUESTION WAS HOW IMPORTANT IS
THIS PROTEIN IN ANCHORING OR
ESTABLISHING THE ENTIRE SUPER
ENHANCER OR ENHANCER COMPLEX?
AND HERE'S A SCHEMATIC,
IDENTIFIED ENHANCER, WHICH IS
VERY COMPLEX BINDING NUMEROUS
PROTEINS, NFIB WHEN I WORKED IN
PHIL'S LAB.
SEVERAL PROTEINS BIND TO DNA,
BUT IT TURNS OUT THAT ONLY ONE
IS THE ANCHOR FOR THIS TTC GAA
MOTIF RECOGNIZED BY STAT5.
THE GENE MUTATES ONE NUCLEOTIDE,
G TO A IN THE MOUSE.
NOT ONLY STAT5 FALLS OFF, BUT
THE ENTIRE LOCUS, ENTIRE
ENHANCER COMPLEX DISAPPEARS,
CANNOT BIND ANYMORE.
SINGLE ANCHOR WHICH RECRUITS
OTHER PROTEINS, WHICH ALSO WOULD
BIND TO DNA, BUT THEY DON'T BIND
TO SPECIFIC MOTIFS.
SO, IN BOSTON, KATHY MENTIONED
ULLI, I ALSO FOUND A MATE,
PRISCILLA WORKED ACROSS THE QUAD
IN BERNIE'S LAB.
IT OF I WENT ON A BLIND -- I ON
A BLIND DATE, KELSEY MIGHT
REMEMBER, THE PIZZA PLACE,
CORRECT?
ACTUALLY PHIL ADOPTED PRISCILLA,
MOLECULAR EXPERIMENTS, MANY
TEST TUBES, JUST SEQUENCE IT,
JUST SEQUENCE IT, REMEMBER THAT?
AND SO WE ALWAYS STOPPED IN
PHIL'S LAB, ON VACATION, THIS IS
THE BUILDING WHERE WE STARTED
IT.
YOU CAN SEE THREE CHILDREN, THE
HUMAN REPRODUCTION.
AND IN HONOR OUR SON ZAK, HIS
MIDDLE NAME IS BOSTON, HE MOVED
BACK TO BOSTON AND PEOPLE DON'T
BELIEVE HIM SHOW HE HAS TO SHOW
HIS DRIVER'S LICENSE.
THOSE WERE THE GOOD OLD DAYS.
YOU MIGHT RECOGNIZE YOURSELF.
JOHN, YOU STILL LOOK THE SAME.
PHIL ALWAYS VERY PROPER.
AYA, YOU CAN SEE MYSELF OVER
THERE.
KATHY, JIM BATTEY ALSO MENTIONED
BY KATHY, AND JAY.
THANK YOU VERY MUCH.
[APPLAUSE]
>> IT'S A SMALL WORLD, ISN'T IT?
OUR NEXT SPEAKER IS THE MUCH
REFERENCED CYNTHIA MORTON.
WILLIAM LAMBERT RICHARDSON
PROFESSOR OF OBSTETRICS AND
GYNECOLOGY, DIRECTOR OF
CYTOGENETICS, PROFESSOR OF
PATHOLOGY HARVARD, UNDERGRAD
FROM COLLEGE OF WILLIAM AND
MARY, Ph.D. FROM MEDICAL
COLLEGE OF VIRGINIA.
SHE WAS A POSTDOC IN THE
LABORATORIES OF PHIL LEDER,
JOINED THE HARVARD FACULTY,
OVERALL THEME IS APPLICATION OF
EVOLVING TECHNIQUES IN MOLECULAR
CYTOGENETICS TO ADDRESS PROBLEMS
IN HUMAN CYTO GENETICS, THERE IS
INCLUDE IDENTIFYING GENES IN
HEARING, DEAFNESS DISORDERS, AS
WELL AS OTHER DEVELOPMENTAL
DISORDERS.
SHE'S BEEN THE PRESIDENT OF THE
AMERICAN SOCIETY OF HUMAN
GENETICS, SERVED NIH IN MANY
CAPACITIES INCLUDING BEING CHAIR
OF THE NIDCD BOARD OF SCIENTIFIC
COUNSELORS, RECIPIENT OF THE
DISTINGUISHED CYTOGENTETICIST
AWARDS AMONG OTHERS.
[APPLAUSE]
OF.
>> GOOD AFTERNOON.
WHAT AMAZING MEMORIES HAVE COME
UP IN ME BEING HERE TODAY.
IT'S REALLY BEEN ENORMOUSLY
SPECIAL.
SO, I HAVE A WARM MEMORY OF THE
FIRST IN-PERSON MEETING WITH
PHIL LEDER AT A RECEPTION IN THE
GARDEN OF THE JERUSALEM MUSEUM
IN THE FALL OF 1981.
I WAS A FIFTH YEAR GRADUATE
STUDENT, IN HUMAN GENETICS,
MEDICAL COLLEGE OF VIRGINIA IN
RICHMOND.
THE STORY IS ONE THAT'S A
FAVORITE IN MY LIFE THAT BEGINS
AT A WORKSHOP AT THE SHORT
COURSE IN EXPERIMENTAL MAMMALIAN
GENETIC, IN BAR HARBOR, CONNECTS
WITH RICK, AMAZING THESE THINGS
HAPPENED IN LIFE.
AS A GRAD STUDENT I HAD CHOSEN
HUMAN CYTOGENETICS AS MY
SPECIALLY.
I VOLUNTEERED TO ASSIST IN
KARYOTYPING WORKSHOP WITH AN
OPPORTUNITY TO CUT PHOTOGRAPHS
OF PRO METAPHASE AND META PHASE,
ALIGN IN SPARES WITH DOUBLE
STICKY TAPE.
THROUGH A CHANCE MEETING AT THE
TABLE, I JOINED, TO ASSIST
STUDENTS I MET LANNY KIRSCH WHO
ATTENDED THE SHORT COURSE IN
1981, I MET JON, KRICKET, AND A
WORK IN PROBLEM WHO BECAME THEIR
DAUGHTER ANIKA.
I LISTENED TO LANNY DESCRIBING
EXCITING EXPERIMENTS UNDERWAY IN
THE LEDER LABORATORY TO MAP
HUMAN GENES.
BY CHROMOSOMAL IN SITU
HYBRIDIZATION WITH DNA LABELED
PROBES.
A MISSING ELEMENT WAS IN THE
EXPERIMENTAL PLAN, SOMEONE TO
READ THE SLIDES.
I WANTED TO OFFER HELP, AND
PROPOSE APPLYING THE METHOD I
WAS USING IN MY DISSERTATION
RESEARCH TO STAIN CHROMOSOMES
WITH A SILVER NITRATE SOLUTION,
AND THEN ILLUMINATE THE
CHROMOSOMES BY USING
FLUORESCENCE MICROSCOPY.
FOLLOWING THE SILVER STAINING,
YOU CAN SEE BY CHANGING THE
BRIGHT FIELD AND FLUORESCENCE
AND SCORE THE SILVER STAINING
AND THEN DETERMINE WHICH
CHROMOSOME IT WAS.
SO, THERE'S SOME OVERLAP IN THE
EXPERIMENTAL METHOD, SILVER
STAIN ON THE ACTIVE NUKE LEOLA
ORGANIZER REGIONS GENES
ILLUMINATED IN THE PHOTOGRAPHIC
EMULSION BUT IT WAS REALLY JUST
DUMB LUCK.
NEVERTHELESS, ONE DAY A BOX OF
SLIDES ARRIVED IN RICHMOND FROM
LANNY, FOR ME TO ANALYZE.
FRANKLY I DIDN'T THINK FOR ONE
MOMENT WHAT GENE THEY MIGHT BE
MAPPING.
LANNY CALLED ME ONE AFTERNOON
AND I WAS AT THE FLUORESCENCE
MICROSCOPE.
I ANSWERED THE PHONE, TO ASK
WHETHER I LOOKED AT THE SLIDES.
I HADN'T.
AFTER ALL, I WAS BUSY WITH MY
OWN DISSERTATION RESEARCH, BUT I
STOPPED AND STAINED THE LIDS --
SLIDES AND WENT TO THE
FLUORESCENT MICROSCOPES.
I KNEW IN AN HOUR THE LOCATION
OF THE DNA PROBE.
LATER THAT AFTERNOON, CAME THE
LIFE-CHANGING PHONE CALL FROM
LANNY.
HE INQUIRED AS TO WHETHER I
COULD DETERMINE THE LOCATION OF
THE SILVER GRAINS, AND I SAID,
YES, 14.
AND THEN HE ASKED IF I COULD
DETERMINE WHERE THEY WERE, ON
CHROMOSOME 14, AND I SAID, OH,
YES, THEY ARE AT 14Q 32.
SO THIS IS A SORT OF FUZZY IMAGE
FROM THE MICROSCOPE WHERE YOU
CAN SORT OF SEE JUST THE
FLUORESCENCE AND THEN THE
FLUORESCENCE WITH THE BRIGHT
FIELD TURNED UP, AND YOU CAN SEE
ON BOTH CHROMOSOMES 14 THEY HAD
THIS LARGE BLACK AREA OF
STAINING.
AND WE PREPARED THESE TYPES OF
IDIOGRAMS FOR PUBLICATION.
THERE WAS NO POWERPOINT AT THAT
POINT.
OKAY.
SO, THERE'S AN EVEN LARGER
VERSION OF WHAT THE CHROMOSOME
14 LOOKED LIKE.
SO, THEN THERE'S ANOTHER PLAYER.
LANNY ASKED IF SOMEONE IN THE
LAB COULD PICK UP THE PHONE AND
JUST ANSWER SOME QUESTIONS THAT
I HAD JUST ANSWERED.
SURE.
THAT SOMEONE WAS JEFF RADISH.
AND SO LANNY REPEATED THE SAME
QUESTIONS.
YOU KNOW, WHAT CHROMOSOME IT'S
ON, DO YOU KNOW WHERE IT IS ON
CHROMOSOME 14.
AND THAT WAS THE MAPPING OF THE
IMMUNOGLOBULIN HEAVY CHAIN
LOCUS.
AND I REMEMBER JUST STANDING IN
THE LAB FOR A LITTLE BIT
THINKING, WHAT JUST HAPPENED?
LITTLE DID I REALIZE THAT THIS
WOULD CHANGE MY ENTIRE CAREER.
AND THEN WE WENT ON TO ACTUALLY
MAP LOTS OF GENES, AND IT WAS
JUST TERRIFICALLY FUN, I'LL TELL
YOU.
SO, THEN ANOTHER SORT OF CHANCE
THING HAPPENED, IN THAT I HAD
ACCEPTED A POSITION AT A POSTDOC
AT HARVARD, BOSTON CHILDREN'S
HOSPITAL, WITH A JUNIOR
INVESTIGATOR THERE, AND THE
LEDER LAB WAS SO EXCITING, IT
WAS REALLY HARD, IT WAS LIKE AT
NIGHT I WOULD GO OVER THERE, AND
DO MORE GENE MAPPING.
EVENTUALLY, I DECIDED THAT THIS
IS WHAT I REALLY WANTED TO DO.
AND I WANTED TO ASK PHIL IF I
COULD JOIN HIS LABORATORY.
SO, I JUST WAS STANDING IN THE
DOOR AND ASKED HIM THIS
QUESTION.
HE SAID, CLOSE THE DOOR AND SIT
DOWN.
OKAY.
AND AFTER I POPPED THAT
QUESTION, HE TOLD ME THAT IT
REALLY WOULDN'T BE GOOD FOR HIM
TO BE RECRUITED TO THE FACULTY
AND FOR HIM TO THEN TAKE THE
POSTDOCS FROM THE JUNIOR
FACULTY.
I SAID, I UNDERSTAND THAT.
BUT I'M NOT GOING TO STAY WHERE
I AM.
I'M TELLING YOU THAT FOR SURE.
AND I KNOW BOSTON HAS LOTS OF
OPPORTUNITIES, SO PROBABLY I'LL
BE LEAVING THAT LAB ANYWAYS.
IT'S JUST THAT THE LAB WAS SUCH
AN EXCITING PLACE, MOST OF THE
FOLKS IN THE CHILDREN'S
DEPARTMENT AT THAT POINT HAD
FAMILIES AND CHILDREN AND THEY
WERE GOING HOME AT NIGHT AND I
WAS THERE BY MYSELF.
AND SO IT REALLY BECAME AT THAT
POINT -- I CAN'T SPEAK FOR WHAT
WAS GOING ON ON THE SIXTH FLOOR,
STU WAS ON THE 6th FLOOR, I
WAS EXCITED TO JOIN THE LAB.
AND THEN THERE WAS THE NEXT
EXPERIMENT, ONE OF THE NEXT
MAPPING EXPERIMENTS, WHICH WAS
THE MYC GENE.
YOU CAN SEE THE SPIKE AT 8Q24.
THIS WAS TRULY A DREAM COME
THROUGH FOR A YOUNG
CYTOGENETICIST, A PATH TO
DISCOVER THE CHROMOSOMAL
REARRANGEMENTS, BUT FOR MYC AND
IMMUNOGLOBULIN THIS REALLY ONLY
TOOK PLACE IN SEPTEMBER --
PUBLICATION IN SEPTEMBER IN
"SCIENCE" LAST YEAR, 2022.
SO, THERE WAS A PAPER THAT
SHOWED THAT REPLICATION TIMING
IS RELATED TO TRANSLOCATION
REARRANGEMENTS, AND THAT
IMMUNOGLOBULIN IS IN THE SAME
TOPOLOGICAL-ASSOCIATED DOMAIN AS
IS MYC.
SO, DOUBLE-STRAND BREAKS OCCUR,
AS YOU KNOW, IN THE ONTOGENY OF
IMMUNOGLOBULIN BIOLOGY, AND THEN
SOME STATES TAKE PLACE IN MYC,
DOUBLE-STRAND BREAKS OCCUR,
THAT'S THE ORIGIN OF THE
TRANSLOCATIONS.
SO IT'S POSSIBLE THAT MAYBE
THERE WOULD BE SOME POTENTIAL
THERAPY BY ACTUALLY PUSHING MYC
INTO A DIFFERENT REPLICATION
TIMING DOMAIN.
I WANTED TO SHARE THAT PAPER
WITH PHIL TO SAY NOW WE KNOW, 39
YEARS LATER, WHICH WENT BY LIKE
A FLASH.
BUT I TOOK THE SECOND BEST IDEA,
WHICH WAS TO GO AND SHARE THIS
WITH AYA.
SO WE SAT FOR HOURS TALKING
ABOUT THIS PAPER.
IT WAS JUST AMAZINGLY FUN, JUST
TOTALLY HAD A GREAT TIME, ABOUT
6 TO 8 WEEKS AGO NOW.
THE LEDER LAB WAS A FAMILY
PLACE.
SOME OF MY HAPPIEST DAYS IN THE
LAB WERE THERE.
THE RULES WERE EASY TO LIVE BY.
I OFTEN ORIENTED NEW MEMBERS OF
THE LAB, TELLING THEM NOT TO PUT
TAPE ON THE WALLS OR FURNITURE
OF THE LABORATORY.
[LAUGHTER]
SO, ONCE WHILE LOOKING OVER MY
SHOULDER, PHIL REMINDED ME THAT
IT WAS POSSIBLE TO USE BOTH ENDS
OF THE TOOTHPICK.
[LAUGHTER]
I SAID, OKAY.
SO, THAT'S JUST BEING THRIFTY,
RIGHT?
THAT WAS A GOOD IDEA.
HE WAS A FABULOUS SPEAKER.
AND THERE WERE TIMES WHEN I
ATTENDED HIS TALKS AT HARVARD
JUST TO WATCH HIM GIVE THE TALK.
AND AT SOME TIMES I FELT THAT HE
WAS GAZING UP FOR DIVINE
INSPIRATION AND THEN WOULD TELL
YOU ABOUT THIS EXPERIMENT.
IT WAS REALLY FABULOUS.
I KNOW THAT THERE ARE MANY
MEMORIES LIKE THIS, AMONG OTHERS
HERE, THAT SHAPED THEIR LIVES,
AND HE LED US ALL BY EXAMPLE.
HIS PASSION FOR SCIENCE, HIS
SKILL AS A TEACHER AND
COMMUNICATOR, THROUGH HIS
HUMANITY, AND DECENCY, HE MADE
US ALL BETTER PEOPLE AND BETTER
SCIENTISTS.
SO, I'M JUST GOING TO WRAP UP
HERE BY THANKING THE ORGANIZERS
OF THIS MEETING FOR GIVING US
THIS OPPORTUNITY TO BE TOGETHER.
AND THANK YOU SO MUCH FOR
CONSTRUCTING THIS HOUSE, WHICH
THERE'S NO DOUBT HE WOULD LOVE
THIS.
HE WOULD ABSOLUTELY LOVE THIS.
SO, I BELIEVE THAT THIS HOME
WILL NOURISH AMONG ITS OCCUPANTS
SOME OF THE CHARACTER TRAITS AND
SCIENTIFIC GENIUS OF A GREAT MAN
WHO LOVED HIS FAMILY.
I HOPE THAT THOSE OF YOU WHO ARE
SO FORTUNATE TO LIVE WITHIN
THESE WALLS, AND I IMAGINE THE
STUDENTS ARE BUSY SO THEY ARE
NOT HERE TODAY, BUT WILL ACQUIRE
AT LEAST A BIT OF HIS REMARKABLE
CHARACTER.
FOR CERTAIN, THERE'S NEVER A DAY
WHEN I'M HAPPIER THAN ONE WHERE
I HAVE AN OPPORTUNITY TO SEIZE
THE MOMENT AND TO SHARE THAT I
WAS A POSTDOC IN THE LEDER
LABORATORY.
HE TRULY SHAPED MY LIFE AND MY
CAREER, AND HE WAS SIMPLY THE
BEST.
[APPLAUSE]
>> OKAY.
FINAL SPEAKER IS RICHARD
WOYCHIK, DIRECTOR OF THE
NATIONAL INSTITUTES OF
ENVIRONMENTAL HEALTH SCIENCES.
RICK DID HIS UNDERGRADUATE WORK
AT UNIVERSITY OF WISCONSIN, GOT
HIS Ph.D. FROM CASE WESTERN
RESERVE UNIVERSITY.
JOINED THE LEDER LAB 1984 TO
1987 AT HARVARD, MOVED TO OAK
RIDGE FOR TEN YEARS, DIRECTING
THE OFFICE OF FUNCTIONAL
GENOMICS, BEFORE RETURNING TO
CASE WESTERN AS A PROFESSOR IN
THE DEPARTMENT OF PEDIATRICS AND
VICE CHAIRMAN FOR RESEARCH
THERE.
HE WAS HEAD OF THE PARKE-DAVIS
LABORATORY FOR MOLECULAR
GENETICS DEVELOPMENT AND LINKS
THERAPEUTICS BEFORE BECOMING
PRESIDENT AND CEO OF THE JACKSON
LABORATORY IN BAR HARBOR.
IN 2011, HE WAS RECRUITED BACK
TO THE NIH AS DEPUTY DIRECTOR OF
NIEHS, AND IN 2020 BECAME THE
DIRECTOR.
HIS RESEARCH PRIOR TO JOINING
NIEHS FOCUSED ON IDENTIFYING
GENES RESPONSIBLE FOR VARIETY OF
HUMAN INHERITED DISORDERS.
RICK.
>> WELL, LET ME START BY SAYING
THAT, AGAIN, IT'S TRULY AN HONOR
TO BE HERE.
PHIL WAS JUST A REMARKABLE
INDIVIDUAL.
SO IF ANY OF YOU TRAINEES IN THE
AUDIENCE OR IF YOU'RE NOT HERE
THEN PASS ON THE WORD, BE VERY
CAREFUL WHO YOU CHOOSE AS YOUR
MENTOR.
YOUR MENTOR CAN HAVE A VERY
POWERFUL INFLUENCE, NOT ONLY ON
THE PERIOD IN WHICH YOU'RE
TRAINING BUT ACTUALLY ON THE
REST OF YOUR CAREER.
I'M GOING TO GIVE YOU A STORY
ABOUT WHAT HAPPENED IN MY
CAREER.
I STARTED OFF IN THE LABORATORY
AS A MOLECULAR BIOLOGIST,
LOOKING AT SOUTHERN BLOTS AND
THEN I ENDED UP AS A MOUSE
GENETICIST.
LET ME TAKE YOU THROUGH WHAT
HAPPENED OVER THE COURSE OF THE
THREE-YEAR PERIOD OF TIME WHEN I
WAS IN THE LEDER LABORATORY FROM
1984 TO 1987.
SO, I ARRIVED IN JUNE OF 1984,
WHEN PHIL HAD JUST -- TIM
STEWART MADE THE TRANSGENIC
MOUSE, MADE THE COVER OF "CELL,"
LINKING THE DYSREGULATED
EXPRESSION OF MYC WITH A
TUMIROGENIC TRAIT IN MAMMARY
TISSUE.
ONE OF MY PROJECTS, I CHOSE TO
CONTINUE TO WORK ON THIS
WONDERFUL MOUSE.
AND I WANTED TO STUDY THE
MOLECULAR ONCOGENESIS AND DO ALL
THE TINES -- TYPES OF THINGS
YOU CAN DO WITH THIS POWERFUL
MOUSE MODEL.
I DISCOVERED THEY WERE WAITING
FOR A POSTDOC TO COME THERE WERE
HUNDREDS OF MICE THAT NEEDED TO
BE GENOTYPED TO DETERMINE
WHETHER THEY CARRIED THE GENE, I
SPENT THE FIRST SEVERAL WEEKS
SNIPPING TAILS IN THE MOUSE ROOM
SORTING THROUGH MICE, SNIPPING
TAILS, MAKING DNA.
AFTER A FEW WEEKS I THOUGHT, IS
THIS WHAT I SIGNED UP FOR?
THERE HAS TO BE AN EASIER WAY.
TAKE A MALE AND FEMALE THAT
CARRY THE GENE AND MAKE A HOMOZY
COAT Z YGOTE, THAT STARTED MY
CAREER IN MOUSE GENETICS.
ONE OF THOSE MMTV MYC MICE
HAD -- WHEN YOU MADE -- 25% OF
OFFSPRING EXHIBIT A LIMB
ABNORMALITY.
IT WAS INTERESTING BECAUSE I
STILL REMEMBER THE SATURDAY
MORNING PHIL WAS IN HIS OFFICE,
I BROUGHT IN THE STAINED
SKELETONS, CHECKING TO SEE
WHETHER THIS NEW MUTATION THAT
AROSE BY INSERTION OF IMMUNOGEN
SIS IN THIS TRANSGENIC LINE WAS
SIMILAR, SIMILAR PHENOTYPE TO
ANY OF THE EXISTING MUTATIONS IN
MARGARET GREEN'S BOOK.
I-- PHIL WAS READING THE
DESCRIPTION OF LIMB DEFORMITY,
INDICATING THERE WAS A FUSION OF
THE RADIUS AND ULNA IN THE
FORELIMB, INTERESTING, THAT'S
WHAT WE'RE SEEING.
THERE WAS A REPLACEMENT OF THE
TIBIA AND FIBULA IN HIND WITH
SINGLE TRIANGULAR BONE.
WOW, THAT'S EXACTLY WHAT WE'RE
SEEING!
AND WENT TO TO DESCRIBE OTHER
ASPECTS OF THE PHENOTYPE.
TURNS OUT THERE WERE SPONTANEOUS
MUTATIONS OF LIMB DEFORMITY AT
THE JACKSON LABORATORY, AND ALSO
OAK RIDGE.
WE ACQUIRED ALLELES, DISCOVERED
WE HAD A NEW MUTATION OF LIMB
DEFORMITY BUT IN OUR CASE THE
MUTATION AROADS BY INSERT OF
MMVT MYC.
REMEMBER, THIS IS IN 1984 AND
85, THERE WERE NO GENOME
SEQUENCES AND THINGS YOU CAN DO.
HAD TO DO POSITIONAL CLONING.
WHAT WE DID ARE OUR ALLELE WAS
USED MYC GENE AS A MOLECULAR
TAG.
WHEN TIM STEWART MADE THE MICE
HE INJECTED ONE OF THE --
REPLICATION ORIGIN AND
ANTIBIOTIC RESISTANT GENE, DID A
PLASMA RESCUE, CLONED THE
FLANKING SEQUENCES.
THAT WAS QUITE REVEALING.
WHAT IT WAS CLEAR TO ME, THIS IS
A GREAT WAY TO CONNECT GENES AND
WE DISCOVERED THIS RATHER NOVEL
GENE ON MOUSE CHROMOSOME 2 WITH
VERY INTERESTING DEVELOPMENTAL
AND HUMAN RELEVANT PHENOTYPES.
AFTER FINISHING THAT CAREER I
DECIDED, I'M GOING TO MAKE A
BUNCH OF MUTATIONS, ONE PLACE
WAS JACKSON LABORATORY, OTHER
WAS OAK RIDGE.
I DECIDED TO GO DOWN, MAKE
HUNDREDS OF LINES OF TRANSGENIC
MICE AND SCREEN FOR INTERESTING
PHENOTYPES.
WE USED PERMACLUDE TECHNIQUE,
WEREN'T DOING HOMOLOGOUS BACK
THEN, NO CRISPR TECHNOLOGY.
WE USED A MINI GENE TO CORRECT
THE ALBINISM DETECT, YOU COULD
TELL IF THE MOUSE HAD A PIGMENT
THE COAT COLOR IT CARRIED THE
GENE, IF IT IT DOUBLE THE DOSE
OR DOUBLE COLOR IN ITS COAT IT
WAS A HOMOZYGOTE.
WE ASSIGNED NUMBERS TO EACH OF
THE ANIMALS, WE GENERATED LOTS
OF DIFFERENT MUTANT LINES.
I COULD TELL YOU ABOUT MANY BUT
I SELECTED ONE TO TELL YOU WHAT
I HOPE YOU'LL FIND AS WE DID A
VERY INTERESTING STORY.
BACK IN THE EARLY 1980s AND
'90s PEOPLE WERE TRYING TO
FIND GENES ASSOCIATED WITH
POLYCYSTIC KIDNEY DISEASE.
WHEN YOU MADE HOMOZYGOTES, THERE
WERE HOLES IN THE KIDNEY, IT
LOOKED LIKE.
IT WASN'T HOLES.
IN THE COLLECTING TUBULES
THERE'S A LOCALIZED
PROLIFERATION OF EPITHELIAL
CELLS, CYSTIC LESIONS.
WE WERE SLICING AND DICING AND
SO IT APPEARED THERE WERE HOLES,
THIS WAS -- WE WORKED WITH A
PATHOLOGIST, THIS WAS A CLASSIC
EXAMPLE OF AUTOSOMAL RECESSIVE
POLYCYSTIC KIDNEY DISEASE, WE
COULD CLONE THE GENE, THIS IS
AMINO ACID SEQUENCE WHICH YOU
CAN'T READ BUT HAD SOME
CHARACTERISTIC FEATURES VERY
INTERESTING, PEPTIDE REPEATS,
SUGGESTING INTERACTION WITH
OTHER PROTEINS.
WE MADE A KNOCKOUT.
THIS IS A FEW YEARS LATER ONCE
TECHNOLOGY WAS AVAILABLE, TO
MAKE CERTAIN THE GENE IS THE ONE
ASSOCIATED WITH PKD.
SO, WHAT WE DID WAS WENT IN AND
DELETED SOME OF THE CODING
SEQUENCES, IF YOU BALANCE THE
INSERTION MUTATION WITH KNOCKOUT
ALLELE YOU HAVE POLYCYSTIC
KIDNEY DISEASE.
WITH THE BETA GALACTIC WE COULD
SEE WHERE IS THE GENE INVOLVED
IN POLYCYSTIC KIDNEY DISEASE
EXPRESSED?
SPECIFICALLY INTERESTED IN
LOOKING AT WHAT EMBRYONIC
DEVELOPMENT.
ON THE LEFT IN PANEL C WE
STAINED FOR LAC C, REFERRED TO
AS THE NODE, ONE OF THE SIGNAL
CENTERS OF THE EMBRYO WITH A
VERY IMPORTANT ROLE IN
LEFT-RIGHT DETERMINATION.
EACH OF THE CELLS IN THE NODE IS
EPITHELIAL CELLS, HAS A
FINGER-LIKE STRUCTURE, THIS
IS -- I'M NOT A DEVELOPMENTAL
BIOLOGIST BUT APPARENTLY THESE
STRUCTURES CREATE SOMETHING
CALLED NODAL FLOW AND THAT
DIRECTS MORPHOGENS TO ALIGN LEFT
AND RIGHT POLARITY IN THE
EMBRYOS.
WE MADE KNOCKOUT MICE HOMOZYGOUS
AND FOUND IT CAUSE THE EMBRYONIC
LETHALITY, STAINED WITH LEFTY 2,
ON PANEL A YOU GET ASYMMETRIC
STAINING BUT IN MUTANT EMBRYOS
GETTING BILATERAL STAINING,
BOTTOM LINE HERE IS THAT THE
INACTIVATION OF THE GENE THAT
CAUSE THE POLYCYSTIC KIDNEY
DISEASE CAUSED LEFT-RIGHT
POLARITY DEFECTS IN THE EMBRYO.
WE DECIDED TO NAME THE GENE
POLARIS.
THAT'S WHERE IT COMES FROM.
IT TOLD US THAT THE ORIGIN
SEGREGATIONS MUTATION WAS A
HYPOMORPHIC ALLELE, WASN'T A
KNOCKOUT, A COMPLETE KNOCKOUT OF
THE GENE.
SO, IT'S VERY INTERESTING.
WHAT'S THE CONNECTION BETWEEN
THE NODE?
CELLS WITH THE CENTRAL PSYLLIUM
AND POLYCYSTIC KIDNEY DISEASE?
THE POSTDOC IS NOW A PROFESSOR
AT UNIVERSITY OF ALABAMA, AND HE
DEVELOPED AN ANTIBODY TO THIS
POLARIS GENE AND ALSO LOOKED AT
THE SURFACE OF THE EPITHELIAL
CELLS AND YOU CAN SEE THIS
PUNCTATE STAINING, IF YOU
STAINED FOR CENTRAL PSYLLIUM YOU
CAN SEE THE CILIATED FINGER-LIKE
STRUCTURE, INVOLVED IN THE
STRUCTURE AND FUNCTION OF THE
CENTRAL CILIUM.
THERE WERE MANY OTHER LABS THAT
CLONED GENES ASSOCIATED WITH
POLYCYSTIC KIDNEY DISEASE, ALSO
COMPONENTS OF THE CENTRAL
CILIUM.
I HAVEN'T FOLLOWED THE PKD WORK.
HOW ARE THESE GENES ASSOCIATED
WITH THE CILIATED STRUCTURE
SOMEHOW CAUSING POLYCYSTIC
KIDNEY DISEASE?
A VERY INTERESTING INSIGHT INTO
GENES AND GENE FUNCTION.
I ALSO WANT TO TALK ABOUT MOUSE
AGOUTI GENE.
A COLLEAGUE AT OAK RIDGE HAD
JUST GENERATED RADIATION INDUCED
MUTATION ON CHROMOSOME 2, THERE
WAS A MUTATION IN LIMB DEFORMITY
AS WELL AS IN AGOUTI GENES.
WE GOT THE COVER OF "CELL" IN
199 1992 BECAUSE AGOUTI WAS A
TARGET OF A TEST AT THE OAK
RIDGE LABORATORY, THEY COULD
DETERMINE WHETHER OR NOT
DIFFERENT PHYSICAL OR CHEMICAL
AGENTED CAUSED MUTATION, A BANK
WAS AVAILABLE TO ANALYZE, THERE
WERE 19 DISTINGUISHABLE ALLELES
OF AGOUTI THAT RANGED FROM
EMBRYONIC LETHALITY TO A COAT
COLOR ALTERATIONS TO ADULT ONSET
OBESITY AND TYPE 2 DIABETES.
SO, A SINGLE GENE GIVING RISE TO
ALL PHENOTYPICALLY PHENOTYPES,
MUST BE A COMPLICATED GENE.
SO WE CLONED THE GENE AND
REMARKABLE THE SIMPLEST, THREE
CODING EXONS ON THE RIGHT, 1
COMES TOGETHER AS MESSAGE RNA
PRODUCES A SMALL SECRETED
PROTEIN WHICH MADE SENSE
RELATIVE TO WHAT WE KNEW ABOUT
HOW AGOUTI WORKS IN THE HAIR
GROWTH CYCLE TO REGULATE
PIGMENTATION OF FUR.
THAT MADE A LOT OF SENSE.
SO, WE WENT ON AND ANALYZED MANY
MUTATIONS, AND IT WAS JUST
REMARKABLE THE WAYS YOU CAN
MUTATE A GENERAL AND TO GIVE
RISE TO THE BROAD ASSORTMENT OF
PHENOTYPES.
SO MAYBE YOU'VE USED C57 BLACK 6
JACK, THIS IS THE STRUCTURE,
SMALL A PANEL HERE.
TURNS OUT IT'S NOT A MUTATION IN
THE CODING SEQUENCE OF AGOUTI,
IT'S A COMPLICATED STRUCTURE.
TWO RETROVIRAL-LIKE ELEMENTS
INSERTED WITHIN THE FIRST INTRON
OF AGOUTI, LTR, LTR-A, AND
ANOTHER VIRAL INSERTION.
THAT'S WHAT BLACK 6 LOOKS LIKE.
SO THIS WAS INTERESTING BECAUSE
IF YOU BRED ENOUGH C57 BLACK 6J
MICE MUTATIONS HAPPEN WITH
REMEMBER LATER, ONE TO BLACK AND
TAN IN THE FIRST SLIDE, MICE
THAT HAVE A YELLOW BELLY AND
BLACK BACK, A FINE LINE ACROSS
THE LATERAL SURFACES.
TURNS OUT WHEN YOU RECOMBINE THE
INSIDE RETROVIRAL ELEMENTS, LTR
B 5 AND 3 PRIME, IF YOU
RECOMBINE OUT THROUGH THE LTR-A,
AND LTRs YOU'RE LEFT WITH SOLO
LTR THAT GIVES RISE TO DIFFERENT
PHENOTYPE WHITE BELLY AGOUTI.
PART OF THE REASON I'M AT THE
NATIONAL INSTITUTE FOR
ENVIRONMENTAL HEALTH SCIENCES IS
IT GOES BACK TO THE ANALYSIS WE
DID ON THIS OTHER VERY
INTERESTING MUTATION.
IT'S A DOMINANT MUTATION THAT
CAUSES YELLOW COAT COLOR, AND
THE MICE THAT HAVE A TOTAL
YELLOW COAT COLOR ALSO HAVE THE
PROPENSITY TO DEVELOP SEVERE
OBESITY AND TYPE 2 DIABETES.
LOWER RIGHT-HAND CORNER IS AN
EXAMPLE OF 85-GRAM MOUSE,
COMPARED TO 25-GRAM LITTER MATE.
DIFFERENCE IN WEIGHT IS ADIPOSE
TISSUE.
SO, WHAT WE WENT ON LEARNING
ABOUT THE AGOUTI GENE, WE
DEMONSTRATED THE MICE HE CAN
TOPICALLY EXPRESS AGOUTI
SIGNALING PROTEIN IN PLACES IT'S
NOT SUPPOSED TO BE NOTABLY IN
THE BRAIN.
WHAT'S HAPPENING IS ATOPIC
EXPRESSION OF THIS SECRETED
PROTEIN IS SHORT CIRCUITING THE
SATIETY RESPONSE, MICE WILL HAVE
LUNCH, THE BRAIN WILL SAY I'VE
HAD ENOUGH TO EAT, I CAN STOP
EATING, BUT IN THIS CASE THE
BRAIN WAS TELLING THEM I'M
STARVING, I'M STARVING, SO THEY
KEEP EATING AND EATING, AND THEY
DEVELOP TYPE 2 DIABETES.
WHAT WAS REALLY INTERESTING IN
OUR CASE IS THAT THE MICE THAT
YOU'RE SEEING, WE MADE THE COVER
OF GENES AND DEVELOPMENT, ALL
MICE ARE GENETICALLY IDENTICAL
AND CARRY THIS AIPY,
PHENOTYPICALLY DIFFERENT.
THEY CARRY THE MUTATION GIVING
RISE TO THESE YELLOW MICE THAT
DEVELOP OBESITY AND TYPE 2
DIABETES.
WE DID THE ANALYSIS.
TURNS OUT THESE ARE CAUSED BY --
INSERTION OF RETROVIRAL
DEVELOPMENT, IAP ELEMENT,
THERE'S A CRYPTIC PROMOTER IN
THE 3 PRIME LTR THAT HIJACKS
NORMAL EXPRESSION.
YOU'RE PRODUCING NOW A FUSION
TRANSCRIPT, CONTAINS ALL OF THE
NORMAL CODING SEQUENCES BUT THE
5 PRIME MESSENGER RNA IS FROM
THE LTR OF THE IAP.
SO WHAT WE ALSO DETERMINED IS
THAT VARIABLE PENETRANCE AND
HIERARCHY IS AAATTRIBUTE USUAL
TOABLE TO OF -- ATTRIBUTABLE TO
EPIGENETICS.
IN MOUSE WITH MODELING, SOME
CELLS LOOK LIKE TOP PANEL AND
SOME LOOK LIKE THE BOTTOM PANEL.
CELLULAR MOSAICS.
HOW DOES ONE GENERATE MICE WITH
VARIABLE PHENOTYPE WHEN IT WAS
INBRED STRAIN?
GENETICS IN NUCLEUS WERE
IDENTICAL.
A POSTDOC CAME UP WITH A GREAT
IDEA THAT, RICK, THEY ARE NOT
TOTALLY IDENTICAL BECAUSE THERE
MAY BE DIFFERENT MITOCHONDRIAL
FUNCTION.
SO IS THERE ROLE FOR DIFFERENT
DEGREES IN MITOCHONDRIAL
FUNCTION IN THE DIFFERENT
INDIVIDUAL ANIMALS?
IS THAT SOMEHOW CONNECTED WITH
THIS VARIABLE PHENOTYPE?
SO WE DECIDED TO TEST THAT.
TURNS OUT THERE'S A GOOD
OLD-FASHIONED NATURAL PESTICIDE
THAT'S DANGEROUS, ROTENONE, THE
TOXICOLOGY COMMUNITY IS
ANALYZING THIS, IT BINDS TO
COMPLEX 1 OF THE ELECTRON
TRANSPORT CHANGE AND TURNS IT
OFF.
AT 10 PARTS PER MILLION WE COULD
TICKLE THE MITOCHONDRIA, CAN WE
INFLUENCE DEGREE OF PENETRANCE
OF THE HIERARCHY?
WE DID THAT.
TESTED THIS DURING DEVELOPMENT.
THE BOTTOM LINE, AGAIN, YOU
HAVE VARIABLE PHENOTYPES ON THE
LEFT, ON THE RIGHT YOU CAN SEE
SPECIFICALLY WITH THE NUMBER OF
MICE THAT ARE FULLY YELLOW AND
HAVE PROPENSITY FOR OBESITY AND
TYPE 2 DIABETES YOU CAN INCREASE
THE NUMBER OF MICE THAT HAVE
THAT PHENOTYPE BY TICKLING THE
MITOCHONDRIA.
WE ASKED MORE RECENTLY IS AGOUTI
AND THIS UNIQUE STRUCTURE OF
AGOUTI THE ONLY GENE THAT
RESPONDS TO THE MITOCHONDRIAL
PERTURBATION OR TREATMENT WITH
PESTICIDES?
OR DOES IT HAPPEN WITH LOTS OF
GENES?
A LOT OF DATA ON HERE BUT THE
BOTTOM LINE WE WENT IN AND DID
WHOLE GENOME BUT SULFIDE
SEQUENCING WITH ZERO PARTS PER
MILL THE EPIGENOME CHANGES OVER
THE COURSE OF TIME, LOOKED AT
DAY 22, SIX MONTHS, 12 MONTHS,
18 MONTHS.
THERE ARE A NUMBER OF DIFFERENT
DIFFERENTIALLY METHYLATED LOCI
ACROSS THE LIFESPAN.
IF YOU LOOK SPECIFICALLY AT THE
LIVES OF THE ANIMALS TREATED
WITH 10 PARTS PER MILLION YOU'LL
SEE THERE IS A MASSIVE
REORGANIZATION OF THE EPIGENOME,
OR THE STATUS, AT 18 MONTHS THEY
PRESERVE THAT ALTERED
METHYLATION PATTERN, OKAY, THAT
WAS ATTRIBUTABLE ONLY TO
ROTENONEEXPOSURE IN DEVELOPMENT.
A BIG REASON I'M AT NIEHS.
HOW DO MANY OF THE ENVIRONMENTAL
TOXICANTS, PESTICIDES, YOU KNOW,
PFAS, HOW ARE THEY INFLUENCED
HUMAN BIOLOGY, AND TO WHAT
EXTENT IS THERE NOW A CONNECTION
WITH ALTERATIONS IN THE
EPIGENOME OF THE MICE.
IN THE INTEREST OF TIME I'LL
FINISH UP BY TALKING ABOUT PHIL.
SO IT'S EXEMPLARY SCIENCE, AN
OUTSTANDING MENTOR.
THERE ARE LOTS OF THINGS I'VE
LEARNED FROM PHIL.
I THINK YOU'VE HEARD THIS FROM
MANY OTHERS.
DON'T BE AFRAID TO SEEK BOLD AN
INNOVATIVE DIRECTIONS.
YOU SHOULD DO THAT.
AND IF NECESSARY, MAYBE INVENT
NEW TECHNOLOGIES, EXPERIMENTAL
APPROACHES TO DO THIS.
OLIGO DT, PHIL INVENTED IT.
WHEN HAD HE TO CLONE A GENE HE
INVENTED THE TECHNOLOGY, I THINK
LAMBDA GT WEST IF I REMEMBER
CORRECTLY.
TREAT ALL YOUR COLLEAGUES WITH
RESPECT.
I WAS SO STRUCK BY MY FIRST HOUR
IN THE LAB, PHIL TOOK ME THROUGH
AND INTRODUCED ME TO SOME NOBEL
LAUREATES, TO POSTDOCS, THE
SUPPORT STAFF WORKING IN THE
DEPARTMENT OF GENETICS.
AND THE ENTHUSIASM OF THIS
INTRODUCTION WAS INDEPENDENT OF
WHERE THEY WERE IN THEIR
SCIENTIFIC CAREER.
THE SUPPORT STAFF WERE
INTRODUCED WITH THE SAME DEGREE
OF ENTHUSIASM AS THE NOBEL
LAUREATES.
SO I THINK I REALIZED, YOU KNOW,
THE MESSAGE IT SENT TO ME
EVERYONE WORKING THERE IS
IMPORTANT AND TREAT PEOPLE WITH
RESPECT.
SO ENCOURAGE YOUR TRAINEES TO
ACHIEVE THE BEST THEY CAN DO TO
ADVANCE THEIR CAREERS.
AND HE WAS ESPECIALLY CAREFUL TO
MAKE SURE IN OUR PRESENTATIONS
WE WERE POLISHED IN THE WAY WE
GAVE OUR TALKS.
SO I STILL THREAP WAS ONE CASE
WHERE ONE OF THE POSTDOCS WAS
TALKING ABOUT RESEARCH AT A LAB
MEETING, EXCUSE MY FRENCH, HE
SAID THERE WAS A SHIT LOAD OF
EXPRESSION IN THE CELL TYPE.
I FELT PALPABLE NERVOUSNESS IN
THE ROOM.
OH MY GOD, PHIL WOULD NEVER TALK
LIKE THAT.
SO, WHEN WE ALL CAME AND GOT OUR
WITS ABOUT IT PHIL SAID, DAN,
DOES THAT MEAN THERE'S A LOT?
ANYWAY, HE HELPED US TO POLISH
THOSE PRESENTATIONS, SO I THINK
LOTHAR TALKED ABOUT PRACTICE
TALKS, MANY OF US HAD THE SAME
EXPERIENCE THE FIRST TIME
THROUGH.
IT WASN'T QUITE AS POLISHED AS
IT ULTIMATELY WAS.
PHIL CARED BUT AS A PERSON.
WHEN I MADE THE DECISION TO DO A
POSTDOC WITH PHIL, I MET
SHIRLEY.
SHIRLEY SAID, GIVE PHIL MY LOVE.
I THOUGHT, THAT'S INTERESTING.
NOW WHEN I LOOK BACK, I REALIZE
THAT I THINK MOST OF US HAVE AN
AFFECTION FOR PHIL THAT GOES
BEYOND AN APPRECIATION FOR HIS
SCIENTIFIC MENTORING.
HE'S A PERSON THAT CARED ABOUT
YOU.
I REALLY CARED ABOUT HIM.
SO IT WAS JUST A REMARKABLE
TIME.
MY FINAL SLIDE IS THIS IS AT THE
EVENT, IN 2004, IT'S -- YOU'RE
LOOKING AT PHIL AND I, IT
REMINDS ME OF THE BEST YEARS OF
MIGHT HAVE SCIENTIFIC CAREER
WHEN I WAS THERE WITH PHIL AND
MANY OF THE REST OF YOU, AT
HARVARD MEDICAL SCHOOL.
SO, THANKS, PHIL.
THANKS, AYA.
SO, IT WAS JUST AN INCREDIBLY
POWERFUL TIME.
SO THANK YOU ALL.
[APPLAUSE]
>> SO, THANK YOU.
THAT WAS QUITE AMAZING.
I HAVE TO SAY AGAIN, TO
REITERATE, QUITE HUMBLING.
THIS SYMPOSIUM HAS BEEN
ABSOLUTELY OUTSTANDING.
THERE HAVE BEEN SOME COMMON
THEMES WE'VE HEARD TODAY.
EDUCATION, COMMUNICATION,
COLLABORATION, MENTORSHIP.
WE ALL THANK PHIL AND FOR MANY
THINGS, FOR HIS CONTRIBUTIONS TO
SCIENCE, TO EDUCATION, ALSO HE
HAS AN AMAZING FAMILY.
THANK YOU, MICKI, ETHAN, BEN.
THANK YOU TO AYA WHO
UNFORTUNATELY COULDN'T BE HERE.
WHAT AN AMAZING FAMILY.
SO WE CAN'T PAY PHIL BACK.
BUT WE CAN PAY IT FORWARD.
FAES IS CONNECTING NIH ALUMNI
AND ALL ACTIVITIES WE DO WHETHER
THROUGH EDUCATION, EVENTS LIKE
THIS, OR NETWORKING EVENTS.
WE'RE WORKING ON SOME NEW
PROGRAMS IN THE NEXT SEVERAL
MONTHS TO YEARS, TO DO THINGS TO
HONOR WONDERFUL PEOPLE LIKE
PHIL, AND WE WOULD LOVE TO HAVE
THE FAMILY BACK.
SO, PLEASE KEEP AN EYE OUT FOR
THE THINGS WE'RE DOING.
WE ARE ALSO VERY EXCITED TO
ALWAYS HAVE NIH-ERS AND OTHER
BIOMEDICAL SCIENTISTS, EXPERTS,
TEACH FOR US AND LECTURE NOW
THAT THINGS ARE ONLINE.
WE CAN DO THINGS REMOTELY AS
WELL.
SO PLEASE REACH OUT.
TAKE A CLASS WITH FAES OR TEACH
A CLASS WITH FAES.
WE WOULD BE VERY HUMBLED TO HAVE
YOU JOIN US.
THINK OF FAES FOR YOUR
EDUCATION.
AND FINALLY, THANK YOU, DR.
GOTTESMAN, FOR WORKING WITH ME
ON THIS PROJECT AND THIS
SYMPOSIUM, LEADING THIS, NOT
ONLY THE LEDER DEDICATION BUT
ALSO THE PROJECT TO BUILD HOUSES
HAS BEEN EXCITING AND HUMBLING
AND IT GIVES FAES AN OPPORTUNITY
TO PAY SOME THINGS BACK, BUT
CERTAINLY RECOGNIZE THE
EXCELLENCE THAT HAPPENS HERE AT
NIH.
SO, THANK YOU FOR BEING HERE.
WE'RE VERY EXCITED.
AND THANK YOU.
WE REALLY APPRECIATE IT.
SOME OF YOU WILL BE JOINING US
FOR THE PHIL LEDER DEDICATION
FROM 2:30 TO 3:30.
WE WILL BE TOASTING PHIL LEDER
AND HIS FAMILY AT THAT EVENT.
THANK YOU AND HAVE A WONDERFUL
DAY.
[APPLAUSE]