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I WOULD LIKE TO WELCOME YOU ALL

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TO THE WEDNESDAY AFTERNOON

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LECTURE SERIES.

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MY NAME IS LAURA KALEY, I AM A

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SENIOR INVESTIGATOR AND BRANCH

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CHIEF OF THE SOCIAL AND

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BEHAVIORIAL RESEARCH BRANCH IN

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THE NATIONAL HUMAN GENOME

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RESEARCH INSTITUTE.

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TODAY'S LECTURE AND SPECIAL FOR

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AT LEAST 2 REASONS, 1 REASON IS

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THAT THIS IS AN NIH MARGARET

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PITTMAN LECTURE WHICH I WILL

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EXPAND UPON BRIEFLY IN A MOMENT

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THE OTHER REASON IS THAT OUR

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SPEAKER DR. GG LOZANO CAN

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FINALLY JOIN US IN PERSON.

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WE NATIONAL  NOMINATED DR. LOZA2

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YEARS AGO, SHE WAS SCHEDULED TO

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SPEAK DURING LAST WALs VISIT

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BUT POSTPONED SO THAT COULD SHE

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COULD DELIVER HER TALK IN THE

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FLESH.

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SO HERE SHE IS, IT'S BEEN REALLY

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EXCITING PLANNING FOR HER VISIT

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AND BUILDING UP FOR THIS LECTURE

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AND WE'RE REAL LE EXCITED TO

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HAVE HER HERE.

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DR. ROTINI WILL INTRODUCE OUR

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SPEAKER TODAY BUT I WANTED TO

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SAY A FEW WORDS ABOUT THIS

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REMARKABLE PERSON, MARGARET

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PITTMAN FOR APPROXIMATE WHOM

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THIS LECTURE IS IN HONOR OF.

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MARGARET PITTMAN CAME TO

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MEDICINE EARLY IN OUR LIFE,

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HELPING HER FATHER WHO WAS A

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COUNTRY DOCTOR IN RURAL ARKANSAS

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AND SHE BECAME SMITTEN WITH

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BIOLOGY AFTER EARNING A

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BACHELOR'S DEGREE AND BIOLOGY

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AND MATHEMATICS SHE WENT INTO

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TEACHING AND SHE USED THE MONEY

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THAT SHE SAVED FROM TEACHING TO

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GO TO THE UNIVERSITY OF CHICAGO

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AND EARNED IN 1928 HER DOCTORATE

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IN BACTERIAERIOLOGY.

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DR. PITTMAN CAME TO THE NIH IN

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1936 BY WAY OF THE ROCKEFELLER

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INSTITUTE AND THE NEW YORK STATE

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DEPARTMENT OF HEALTH.

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AT THE NATIONAL INSTITUTES OF

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HEALTH, HER CAREER PATH LED HER

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TO BECOME THE VERY FIRST WOMAN

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TO HOLD THE POSITION OF

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LABORATORY CHIEF, HEADING THE

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LABORATORY OF BACTERIAL PRODUCTS

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DIVISION OF BIOLOGICS AND

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STANDARDS FROM 1957 TO 1971.

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DR. PITTMAN ISOLATED THE

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INFLUENZA STRAIN, RESPONSIBLE

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FOR MOST CHILDHOOD MENINGITIS,

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AND HELPED IDENTIFY THE CAUSE OF

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EPIDEMIC CONJUNCTIVITIS AND MADE

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KEY OBSERVATIONS THAT LED TO THE

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DEVELOPMENT OF THE SALMONELLA

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VACCINE.

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IN 1970, MARGARET PITTMAN WAS

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RECOGNIZED WITH THE FEDERAL

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WOMEN'S AWARD AND SHE SERVED AS

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THE PRESIDENT OF THE SOCIETY OF

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AMERICAN BACTERIAERIOLOGYISTS

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AND OF THE WASHINGTON ACADEMY OF

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SCIENCES.

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ALTHOUGH DR. PITTMAN RETIRED IN

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1971, SHE KEPT WORKING AT THE

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NIH AS A GUEST UNTIL 1993.

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THUS, SHE CONTRIBUTED TO THE

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SCIENTIFIC COMMUNITY FOR ALMOST

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50 YEARS.

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THE MARGARET PITTMAN LECTURESHIP

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CREATED IN 19 HONORS DR. PITTMAN

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HONORS HER ACHIEVEMENTS AT THE

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NATIONAL INSTITUTES OF HEALTH.

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AS WELL THE MARGARET PITTMAN

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LECTURE HONORS A RESEARCHER WHO

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EXEMPLIFIES THE INTELLIGENCE

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SCIENTIFIC EXCELLENCE AND DRIVE

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OF DR. PITTMAN AND WHOSE

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DEDICATED TO ADVANCING AND

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IMPROVING THE CAREERS OF WOMEN

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SCIENTISTS.

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DR. LOZANO IS MOST CERTAINLY

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DESERVING OF THIS HONOR.

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SO ON BEHALF OF THE WOMEN

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SCIENTIST ADVISORS, I WOULD LIKE

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TO WELCOME DR. GG LOZANO TO THE

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NIH AND WE ARE REALLY LOOKING

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FORWARD TO HEARING MORE ABOUT

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YOUR WORK.

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WITH THAT I WOULD LIKE TO

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INTRODUCE DR. CHARLD ROTINI WHO

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WILL INTRODUCE DR. GIGI LOZANO.

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>>THANK YOU SO MUCH FOR
THAT

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WONDERFUL COMEBTARY ABOUT

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MARGARET PITTMAN.

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IT IS INDEED MY HONOR TODAY TO

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INTRODUCE OUR SPEAKER BUT BEFORE

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I DO THAT, I--MY NAME IS

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DR. ROTINI, AND I AM A MEMBER OF

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THE NATIONAL WHOLE GENOME

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INSTITUTE.

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I HAVE QUICK ANNOUNCEMENTS, 1

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HAS TO DO WITH CONTINUING

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MEDICAL EDUCATION, THE CODE IS

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44434. 

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AND I REPEAT, 44434.

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OKAY, SO, PLEASE SIGN UP AND GET

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YOUR CREDIT FOR ATTENDING THIS

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WONDERFUL TALK.

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ALSO, THERE'S A FEW WATCHING

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REMOTELY, VIA THE NIH VIDEOCAST.

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YOU CAN SEND LIVE FEEDBACK TO

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THE SPEAKER, SIMPLY CLICK ON THE

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BUTTON THAT SAYS SEND LIVE FEED

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WHICH IS BELOW THE VIDEOCAST

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WINDOW.

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TYPE IN YOUR MESSAGE AND WE WILL

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RELATE IT TO OUR SPEAKER AT THE

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END OF HER LECTURE TODAY.

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NOW, MY TREMENDOUS HONOR TO

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INTRODUCE OUR GUEST DR. LOZANO

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IS THE HERBERT L. OLIVE STRINGER

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DISTINGUISHED CHAIR IN ONCOLOGY

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IN HONOR OF SUE [INDISCERNIBLE]

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STRINGER AT MD ANDERSON.

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'S RENOWNED GENETICIST, SHE'S

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RECOGNIZED FOR HER STUDIES OF

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THE P53 TUMOR IN THE PATHWAYS

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WHICH IS ON THE MIND IN THE

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LARGE PERCENT OF HUMAN CANCER

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VIA MUTATIONS AND DELETIONS OF

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P53.

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USING MASS MODEL DR. LOZANO AND

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HER TEAM IDENTIFIED P53 PROTEINS

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AS A TRANSCRIPTIONAL ACTIVATOR.

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THEY SHOWED THAT MDM2 AND MD

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M PROTEINS ARE INHIBITOR OF 13,

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AND OVER TIME OUR LAB STUDIED

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MULTIPLE MUTATIONS AND

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EXTENSIVELY CHARACTERIZING THE

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EFFECT ON THE TUMOR SUPPRESSION

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AND TUMOR REGENESIS.

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HER GROUND BREAKING WORK HAS

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PROVIDED INVALUABLE TESTING

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NOVEL THERAPEUTIC ISHT VENTIONS,

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DR. LOZANO RECEIVED HER

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UNDERGRADUATE DEGREE IN BIOLOGY

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FROM THE UNIVERSITY OF TEXAS RIO

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GRAND VALLEY, FORMERLY THE PAN

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AMERICAN UNIVERSITY.

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SHE RECEIVED HER Ph.D. IN

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BIOCHEMISTRY FROM ROUGH ATOM

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CHESTER UNIVERSITY CONDUCTED

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POST DOCTORAL TRAINING AT

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PRINCETON UNIVERSITY.

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I SHOULD NOTE THAT GIGI,  THAT'S

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WHAT WE CALL HER, LET'S JUST SAY

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THAT SHE SPENT SOMETIME HERE AT

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THE NIH AND THAT IS WHERE WE

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THINK SHE RECEIVED THE BUG OF

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WHAT IS CALLED THE SCIENTIFIC

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BUG AND THE RESEARCH BUG AND SHE

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WAS AT NCI [INDISCERNIBLE] AND I

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WAS JUST TALKING TO MICHAEL

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GOTTESMAN, HE REMEMBERS ALL OF

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THE STORY ABOUT GIGI AND

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INTERRUPTION WITH NCI SO

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HOPEFULLY AT SOME POINT WE CAN

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TALK ABOUT THAT.

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SO, JUST TO COMPLETE THE

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INTRODUCTION, DR. LOZANO HAS

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PICKED UP MANY ACCOLADES AND

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HONORS ALONG THE WAY, SHE IS A

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MEMBER OF THE AMERICAN RESEARCH

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ACADEMY AND MEMBER OF THE

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ACADEMY ARTS AND SCIENCES,

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NATIONAL ACADEMY OF SCIENCES AND

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NATIONAL ACADEMY OF MEDICINE AND

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RECIPIENT OF THE LEADERSHIP ARK

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WARD FOR ADVANCING WOMEN AND

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MINORITY FACULTY AT MD CANCER

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CENTER.

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IN 2020 DR. LOZANO 1 OF THE MOST

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INSPIRING, HISPANICS, LATINO

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SCIENTISTS IN AMERICA.

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THIS YEAR DR. LOZANO,

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[INDISCERNIBLE] A WORD FOR

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FACULTY LEADERSHIP AT MD

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ANDERSON AND I WOULD BE VERY

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REMISS NOT TO SAY THAT AT A

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PERSONAL LEVEL AS A SCIENTIFIC

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DIRECTOR OF NHGRI, DR. LOZANO

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STAYS ON OUR BOARD OF SCIENTIFIC

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COUNSELORS AND WE HAVE JUST

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TAPPED HER SERVICES AGAIN TO BE

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ON OUR BLUE RIBBON PANEL.

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SO THE TITLE OF HER TALK TODAY

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IS MUTANT P53 ACTIVITIES IN

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MOUSE TUMOR MODELS.

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GIGI, WELCOME BACK TO THE NIH.

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>>THANK YOU CHARLES FOR
THAT

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WONDERFUL INTRODUCTION.

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IT'S GREAT TO BE BACK.

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I REALLY--NIH WAS MY VERY FIRST

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LAB EXPERIENCE.

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I LEARNED MOLECULAR BIOLOGY HERE

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AND I THINK I'M JUST VERY GLAD

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TO BE BACK APPROXIMATE SEE SOME

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OLD FRIENDS AND TO TO  FRIENDS U

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FOR YOUR CONTRIBUTIONS TO MY

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CAREER.

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SO TODAY I WILL TALK ABOUT OUR

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MODELS AND I CHANGED THE TITLE

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JUST A LITTLE BIT TO INCLUDE

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SOME WILD-TYPE P53 ACTIVITIES

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BECAUSE, WE'RE TRYING TO REALLY

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UNDERSTAND NOT JUST WHAT

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WILD-TYPE PPEAF DOES BUT WHAT

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HAPPENS WHAT WHEN IT'S NOT THERE

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AND WHAT OTHER ACTIVITIES MUTANT

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P53 MIGHT HAVE.

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SO I THINK THIS WILL WORK.

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OKAY, THESE ARE MY DISCLOSURES.

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SO I'M GOING TO FIRST INTRODUCE

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THE P53 PATHWAY.

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HOPEFULLY--I WONDER IF I HAVE AN

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ARROW?

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PROBABLY NOT.

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OKAY.

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SO THE P53 PROTEIN IS PRESENT AT

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VERY LOW LEVELS IN CELLS, IT'S

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HARDLY DETECTABLE, BUT STRESS,

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DNA DAMAGE, AND INAPPROPRIATE

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ACTIVATION OF AN ONCA GENE WILL

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UNLEASH A SERIES OF KINASES THAT

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PHOSPHORYLATE P53 AND MAKE IT A

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VERY STABLE PROTEIN.

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AND THEN IN TURN P53 FUNCTIONs

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AS A TRANSCRIPTION FACTOR,

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TETRAMERIC TRANSCRIPTION FACTOR

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THAT ACTIVATES HUNDREDS OF GENES

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AND I ONLY SMOA SOME OF THEM ON

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THIS SLIDE AND IN THESE GENES

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HAVE IMPORTANT CELLULAR

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FUNCTIONS.

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PROBABLY THE FIRST 1 THAT WAS

258
00:10:37,000 --> 00:10:40,760
CHARACTERIZED WAS THE ABILITY TO

259
00:10:40,760 --> 00:10:42,480
INITIATE APOPTOSIS, SO P53 CAN

260
00:10:42,480 --> 00:10:44,000
KILL THE CELL, IT CAN ALSO KILL

261
00:10:44,000 --> 00:10:50,520
THE CELL BY ANOTHER PROCESS

262
00:10:50,520 --> 00:10:52,560
CALLED FERROPITOSEIS, BUT IT CAN

263
00:10:52,560 --> 00:10:54,840
ARREST THE CELL, SO IT CAN CAUSE

264
00:10:54,840 --> 00:10:58,800
THE CELL TO ARREST OR UNDERGO

265
00:10:58,800 --> 00:10:59,280
SENESCENSEL.

266
00:10:59,280 --> 00:11:00,080
THIS CELL PSYCHE ALTHOUGH REST

267
00:11:00,080 --> 00:11:04,360
IS NOT A DEAD END, SENESCENSEL

268
00:11:04,360 --> 00:11:06,360
IS A DEAD END.

269
00:11:06,360 --> 00:11:08,440
IN ADDITION, P53 HAS ABILITY TO

270
00:11:08,440 --> 00:11:10,720
REGULATE ITS OWN LEVEL SO

271
00:11:10,720 --> 00:11:12,400
THERE'S WAYS TO ACTIVATION TO

272
00:11:12,400 --> 00:11:13,640
LEAD TO SURVIVAL OF THE CELL AND

273
00:11:13,640 --> 00:11:16,560
IT'S THROUGH THE PROTEIN MDM2,

274
00:11:16,560 --> 00:11:18,720
IT'S A P53 TARGET.

275
00:11:18,720 --> 00:11:23,960
AND IN TURN MDM2 ENCODES LIGASE

276
00:11:23,960 --> 00:11:25,160
THAT TARGETS P53 FOR

277
00:11:25,160 --> 00:11:25,480
DEGRADATION.

278
00:11:25,480 --> 00:11:27,720
SO I THINK WITH THIS MY OPENNIC

279
00:11:27,720 --> 00:11:29,280
VIEW OF THE P53 PATHWAY, CAN YOU

280
00:11:29,280 --> 00:11:31,360
BEGIN TO SEE THAT P53 HAS SHOW

281
00:11:31,360 --> 00:11:33,440
MANY CHOICES IN THE CELL, IT CAN

282
00:11:33,440 --> 00:11:34,760
KILL THE CELL, STOP THE CELL AND

283
00:11:34,760 --> 00:11:36,600
SAY TIME TO GO ON.

284
00:11:36,600 --> 00:11:39,440
AND EVEN THOUGH WE KNOW SOME OF

285
00:11:39,440 --> 00:11:41,200
THE CIRCUMSTANCES WHERE P53

286
00:11:41,200 --> 00:11:42,880
MAKES THESE DECISIONS, MOST OF

287
00:11:42,880 --> 00:11:45,840
THE TIME, WE CAN'T GUESS WHAT

288
00:11:45,840 --> 00:11:50,440
P53 IS GOING TO DO IN THE CELL.

289
00:11:50,440 --> 00:11:55,480
SO THE FIRST FEW SLIDES BECAUSE

290
00:11:55,480 --> 00:11:58,680
WE WERE INTERESTED IN TRYING TO

291
00:11:58,680 --> 00:11:59,600
UNDERSTAND WHAT THE TARGETS ARE

292
00:11:59,600 --> 00:12:01,560
AND COULD THEY HAVE POTENTIAL

293
00:12:01,560 --> 00:12:05,160
ROLES AS TUMOR SUPPRESSORS IN

294
00:12:05,160 --> 00:12:06,040
AND OF THEMSELVES.

295
00:12:06,040 --> 00:12:08,440
IT'S BEEN VERY HARD TO REGRIEWS

296
00:12:08,440 --> 00:12:09,760
P53S INTO THE TUMORS AND THE

297
00:12:09,760 --> 00:12:13,160
IDEA WAS MAKE WE CAN REACTIVATE

298
00:12:13,160 --> 00:12:16,040
AN IMPORTANT DOWN STREAM TUMOR

299
00:12:16,040 --> 00:12:16,440
SUPPRESSIVE PATHWAY.

300
00:12:16,440 --> 00:12:18,800
SO I HAVE TO ALSO INTRODUCE THE

301
00:12:18,800 --> 00:12:19,760
MDM2 MOUSE BECAUSE THIS IS A

302
00:12:19,760 --> 00:12:22,880
TOOL THAT WE USE IN THE LAB, SO

303
00:12:22,880 --> 00:12:27,280
DILUTION OF MDM2 IS AN

304
00:12:27,280 --> 00:12:28,520
EMBRYO--WHOOPS, WRONG SLIDE.

305
00:12:28,520 --> 00:12:31,720
DELETION OF MDM2 IS EMBRYO

306
00:12:31,720 --> 00:12:32,320
LETHAL.

307
00:12:32,320 --> 00:12:33,720
OOPS, I'M PRESSING THE WRONG

308
00:12:33,720 --> 00:12:33,920
BUTTON.

309
00:12:33,920 --> 00:12:35,320
I THOUGHT I WAS PRESSING THE

310
00:12:35,320 --> 00:12:37,480
RIGHT BUTTON.

311
00:12:37,480 --> 00:12:40,800
OKAY, MDM2 LOSS LEADS TO EMBRYO

312
00:12:40,800 --> 00:12:43,520
LEATHAL PHENOTYPE AT THE BLAST

313
00:12:43,520 --> 00:12:45,880
SIS STAGE, THIS CELL IS

314
00:12:45,880 --> 00:12:46,600
UNDERGOING APOPTOSIS, EVERY 1 OF

315
00:12:46,600 --> 00:12:47,800
THOSE CELLS IS DEAD AND WE ASKED

316
00:12:47,800 --> 00:12:49,360
THE QUESTION WHAT IS THE ROLE OF

317
00:12:49,360 --> 00:12:52,040
PPEAR IN THIS PROCESS AND IF WE

318
00:12:52,040 --> 00:12:54,360
DELETE P53, WE GET A PERFECTLY

319
00:12:54,360 --> 00:12:56,440
VIABLE MOUSE, SO THIS REALLY

320
00:12:56,440 --> 00:12:58,080
CEMENTED THE RELATIONSHIP

321
00:12:58,080 --> 00:12:58,760
BETWEEN MDM2 AND PPARADOXICAL

322
00:12:58,760 --> 00:13:00,440
AND IN FACT WE NOW HAVE

323
00:13:00,440 --> 00:13:02,080
CONDITIONAL ALLELES AND WE'VE

324
00:13:02,080 --> 00:13:05,240
TAKEN MDM2 OUT IN ANY TISSUE, IT

325
00:13:05,240 --> 00:13:07,400
IS ALWAYS A CELL LETHAL AND IT'S

326
00:13:07,400 --> 00:13:09,720
ALWAYS IN A P53 DEPENDENT

327
00:13:09,720 --> 00:13:09,960
FASHION.

328
00:13:09,960 --> 00:13:12,280
SO WHAT WE'VE DONE THEN IS WE'VE

329
00:13:12,280 --> 00:13:14,360
USED A MODEL IN WHICH WE USE

330
00:13:14,360 --> 00:13:22,920
TISSUE AND M

331
00:13:22,920 --> 00:13:23,440
ACROPHAGES--TAMOXIFEN AND

332
00:13:23,440 --> 00:13:25,040
WITHIN A FEW DAYS OF INJECTION,

333
00:13:25,040 --> 00:13:26,360
ALL THE ANIMALS ARE DEAD SHOWN

334
00:13:26,360 --> 00:13:30,600
HERE BY THE RED LINE AND AND

335
00:13:30,600 --> 00:13:32,280
THIS IS A P53 DEPENDENT PROCESS

336
00:13:32,280 --> 00:13:35,680
BECAUSE IF YOU REMOVE P53, THE

337
00:13:35,680 --> 00:13:37,120
ANIMALS ARE PERFECTLY VIABLE.

338
00:13:37,120 --> 00:13:40,240
SO THIS ALLOWS US THEN TO ASK,

339
00:13:40,240 --> 00:13:42,800
WHAT HAPPENS--WHAT IS THE P53

340
00:13:42,800 --> 00:13:44,440
PHYSIOLOGICAL RESPONSE UPON MDM2

341
00:13:44,440 --> 00:13:44,760
DELETION.

342
00:13:44,760 --> 00:13:47,320
AND WE DID THIS IN A SIMILAR

343
00:13:47,320 --> 00:13:50,000
MANNER, BUT IN AN ACUTE WAY SO

344
00:13:50,000 --> 00:13:53,840
THAT 24 HOURS AFTER THE LAST

345
00:13:53,840 --> 00:13:55,120
INJECTION OF TAMOXIFEN IN WHICH

346
00:13:55,120 --> 00:13:57,000
WE HAVE 50% OF THE CELLS THAT

347
00:13:57,000 --> 00:14:00,440
HAVE LOST MDM2 WE CAN EVALUATE

348
00:14:00,440 --> 00:14:01,160
THAT TRANSCRIPTIONAL PROGRAM.

349
00:14:01,160 --> 00:14:02,080
AND THE FIRST THING WE DID WAS

350
00:14:02,080 --> 00:14:04,480
JUST LOOK AT THE PATHOLOGIES OF

351
00:14:04,480 --> 00:14:05,920
SEVERAL OF THE TISSUES IN THE

352
00:14:05,920 --> 00:14:10,160
MOUSE, AND JUST 24 HOURS AFTER

353
00:14:10,160 --> 00:14:11,680
RECOMBINING AND LOSING MDM2, WE

354
00:14:11,680 --> 00:14:13,400
HAVE ALL THESE FAZINATING

355
00:14:13,400 --> 00:14:14,880
PATHOLOGIES, THE HEART AND THE

356
00:14:14,880 --> 00:14:19,640
OVARY, I'M PUSHING THE RIGHT

357
00:14:19,640 --> 00:14:22,680
BUTTON--THE HEART AND THE OVARY

358
00:14:22,680 --> 00:14:23,840
ARE PERFECTLY NORMAL, THEY DON'T

359
00:14:23,840 --> 00:14:25,040
SHOW ANY TIME POINTSA THE THIS

360
00:14:25,040 --> 00:14:28,360
POINT BUT CAN YOU SEE THE

361
00:14:28,360 --> 00:14:29,040
INTESTINE, KIDNEY, PAN KRIST AS

362
00:14:29,040 --> 00:14:31,000
HAVE ALL TYPES OF PHENOTYPES.

363
00:14:31,000 --> 00:14:33,280
IN THE INTESTINE WE SEE LOSS OF

364
00:14:33,280 --> 00:14:38,520
THE CRIPS, IN THE KIDNEY WE SEE

365
00:14:38,520 --> 00:14:39,880
DILATED TUBULES AND SUGGESTING

366
00:14:39,880 --> 00:14:42,600
DYSFUNCTION THERE AND IN THE

367
00:14:42,600 --> 00:14:46,560
PANCREAS WE SEE THIS SAR

368
00:14:46,560 --> 00:14:47,240
METADUCTILE TRANSPLACIA.

369
00:14:47,240 --> 00:14:49,320
SO THEN WE LOOK AT ADDITIONAL

370
00:14:49,320 --> 00:14:51,760
TARGETS OF P53 AND WE IDENTIFIED

371
00:14:51,760 --> 00:14:53,480
HUNDREDS OF TARGETS SO FOR

372
00:14:53,480 --> 00:14:56,480
EXAMPLE IN THE PANCREAS, WE

373
00:14:56,480 --> 00:14:57,800
IDENTIFIED 206 P53 TARGETS.

374
00:14:57,800 --> 00:15:00,600
IN THE OVARY WHICH HAD NO

375
00:15:00,600 --> 00:15:01,280
PHENOTYPE, WE IESHES DENTIFIED

376
00:15:01,280 --> 00:15:03,880
21 TARGETS AND THESE ARE P53

377
00:15:03,880 --> 00:15:05,040
DIRECT TARGETS BECAUSE WE

378
00:15:05,040 --> 00:15:09,160
BASICALLY TOOK THE RNA SEQ DATA

379
00:15:09,160 --> 00:15:11,800
AND COMBINED IT WITH CHPSEQ DATA

380
00:15:11,800 --> 00:15:12,720
THAT WAS AVAILABLE IN THE

381
00:15:12,720 --> 00:15:16,080
LITERATURE AND THIS IS THE P53

382
00:15:16,080 --> 00:15:18,520
TRANSCRIPTIONAL PHYSIOLOGICAL

383
00:15:18,520 --> 00:15:18,760
PROGRAM.

384
00:15:18,760 --> 00:15:20,080
SO I THINK THERE ARE 2 IMPORTANT

385
00:15:20,080 --> 00:15:22,040
POINTS I WANT TO MAKE, THERE ARE

386
00:15:22,040 --> 00:15:23,680
ONLY 7 GENES THAT ARE COMMON

387
00:15:23,680 --> 00:15:25,960
BETWEEN THESE 5 TISSUES AND MDMD

388
00:15:25,960 --> 00:15:27,000
IS 1 OF THEM.

389
00:15:27,000 --> 00:15:29,800
SO IT'S INTERESTING THAT MDM2 IS

390
00:15:29,800 --> 00:15:31,440
REGULATED BY P53 IN THE

391
00:15:31,440 --> 00:15:32,160
PHYSIOLOGICAL SYSTEM IN EVERY

392
00:15:32,160 --> 00:15:35,200
TISSUE THAT WE ANALYZED.

393
00:15:35,200 --> 00:15:39,040
BUT IT'S PROBABLY DOING IS

394
00:15:39,040 --> 00:15:40,480
BALANCING THAT P53 LEVELS, MAYBE

395
00:15:40,480 --> 00:15:47,160
AT DIFFERENT TIMES DURING THE

396
00:15:47,160 --> 00:15:47,520
CELL CYCLE.

397
00:15:47,520 --> 00:15:51,160
POLK IS A POLYMERASE, EDA2 R IS

398
00:15:51,160 --> 00:15:52,240
A CELL SURFACE RECEPTOR INVOLVED

399
00:15:52,240 --> 00:15:53,000
IN CELL DEATH.

400
00:15:53,000 --> 00:15:54,520
BUT TO ME THE OTHER FASCINATING

401
00:15:54,520 --> 00:15:57,520
ASPECT OF THIS SLIDE IS HOW MANY

402
00:15:57,520 --> 00:15:59,040
TARGETS ARE TISSUE SPECIFIC.

403
00:15:59,040 --> 00:16:05,200
SO FOR EXAMPLE, AGAIN IN THE

404
00:16:05,200 --> 00:16:05,840
PANCREAS, 135 TRANSCRIPTIONAL

405
00:16:05,840 --> 00:16:06,440
TARGETING SYSTEM EXPOSURE TO

406
00:16:06,440 --> 00:16:08,280
RADIATIONS ARE ONLY EXPRESS

407
00:16:08,280 --> 00:16:09,400
INDEED THE PANCREAS, SO THAT

408
00:16:09,400 --> 00:16:11,440
TELLS YOU THERE'S A HUGE

409
00:16:11,440 --> 00:16:12,880
DIVERSITY OF TRANSCRIPTIONAL

410
00:16:12,880 --> 00:16:14,440
TARGETS THAT P53 CAN REGULATE.

411
00:16:14,440 --> 00:16:16,040
WE'VE DONE 1 ADDITIONAL

412
00:16:16,040 --> 00:16:17,520
VALIDATION EXPERIMENT TO ASK IF

413
00:16:17,520 --> 00:16:20,760
THIS IS A TRUE P53 RESPONSE BY

414
00:16:20,760 --> 00:16:22,400
ERADIATING THE WHOLE MOUSE AND

415
00:16:22,400 --> 00:16:23,520
ISOLATING THESE SAME TISSUES,

416
00:16:23,520 --> 00:16:25,880
AND YOU CAN SEE HERE IN THE

417
00:16:25,880 --> 00:16:28,320
MIDDLE IS A WILD-TYPE MOUSE THAT

418
00:16:28,320 --> 00:16:31,920
WAS RADIATED, SO THOSE ARE THE

419
00:16:31,920 --> 00:16:35,400
SAMPLES THAT THE TRANSCRIPTIONAL

420
00:16:35,400 --> 00:16:37,200
TARGETS--OKAY, I'M NOT USING THE

421
00:16:37,200 --> 00:16:38,280
POINTER ANYMORE.

422
00:16:38,280 --> 00:16:41,080
SO THAT MIDDLE PORTION IS

423
00:16:41,080 --> 00:16:43,200
WILD-TYPE P53 AFTER RADIATION

424
00:16:43,200 --> 00:16:45,920
AND THOSE ARE THE LIKE LYNN G

425
00:16:45,920 --> 00:16:47,920
AND ED82 R TARGETS THAT ARE

426
00:16:47,920 --> 00:16:49,800
UPREGULATED IN A WILD-TYPE

427
00:16:49,800 --> 00:16:51,720
ERADIATED MOUSE BUT NOT IN A P53

428
00:16:51,720 --> 00:16:52,120
NULL MOUSE.

429
00:16:52,120 --> 00:16:55,480
SO THIS IS NOT JUST--THIS IS THE

430
00:16:55,480 --> 00:16:57,120
TRANSCRIPTIONAL PROGRAM, WITH

431
00:16:57,120 --> 00:17:02,960
LOSS OF MDM2 AND IN THE PRESENCE

432
00:17:02,960 --> 00:17:03,320
OF RADIATION.

433
00:17:03,320 --> 00:17:05,200
SO I THE OTHER MAJOR FINDING WAS

434
00:17:05,200 --> 00:17:12,320
REALLY WHEN WE TRIED TO ANALYZE

435
00:17:12,320 --> 00:17:13,840
THIS ASENAR PHENOTYPE IN THE

436
00:17:13,840 --> 00:17:14,120
FACIA.

437
00:17:14,120 --> 00:17:16,040
BECAUSE AFTER 24 HOURS WE SEE A

438
00:17:16,040 --> 00:17:18,480
COMPLETE SORT OF REWIRING OF THE

439
00:17:18,480 --> 00:17:24,560
PANCREAS FROM NORMAL AT THE TOP

440
00:17:24,560 --> 00:17:26,200
LEFT TO THIS DUCTILE STRACTURE

441
00:17:26,200 --> 00:17:28,840
AND WE STAINED WITH THE DUCTILE

442
00:17:28,840 --> 00:17:30,480
STAINING YOU CAN SEE THE BROWN

443
00:17:30,480 --> 00:17:32,760
STAINING SO YOU CAN SEE A

444
00:17:32,760 --> 00:17:33,480
PLASTICITY OF TRANSITION THAT

445
00:17:33,480 --> 00:17:34,600
OCCURS AND THE DATA ARE

446
00:17:34,600 --> 00:17:35,360
QUANTIFIED ON THE RIGHT.

447
00:17:35,360 --> 00:17:37,080
SO THE QUESTION WE WANTED TO

448
00:17:37,080 --> 00:17:39,800
ASK, WE WANTED TO DEVELOP A

449
00:17:39,800 --> 00:17:41,640
MODEL THAT WE WERE JUST LOOKING

450
00:17:41,640 --> 00:17:43,520
AT THIS TRANSITION IN THE

451
00:17:43,520 --> 00:17:48,880
PANCREAS AND WE USED AN SNR

452
00:17:48,880 --> 00:17:50,760
SPECIFIC KREE RECOMBIN ACE, THIS

453
00:17:50,760 --> 00:17:55,000
IS A--THIS IS MST 1 CREE ER

454
00:17:55,000 --> 00:17:57,840
TRANSGENE SO WE ADD TAMOXIFEN

455
00:17:57,840 --> 00:17:59,960
AND WE ASKED OURSELVES DO YOU

456
00:17:59,960 --> 00:18:01,480
SEE DELETION OF MDM2.

457
00:18:01,480 --> 00:18:04,200
WHEN WE COMBEAR THE

458
00:18:04,200 --> 00:18:04,840
RECOMBINATION FREQUENCIES FROM

459
00:18:04,840 --> 00:18:05,960
FIRST MOUSE AND THIS MOUSE, THEY

460
00:18:05,960 --> 00:18:08,520
ARE SIMILAR SO WE ARE DELETING

461
00:18:08,520 --> 00:18:11,480
MDM 2 AND ACTIVATING P53 IN THE

462
00:18:11,480 --> 00:18:13,160
PANCREAS.

463
00:18:13,160 --> 00:18:16,760
BUT WE COMPLETELY LOST THIS SNR

464
00:18:16,760 --> 00:18:19,040
TO DUCTILE METAPLACIA, SO NOW

465
00:18:19,040 --> 00:18:21,160
THE TUMOR LOOKS--NOT TUMOR, THE

466
00:18:21,160 --> 00:18:22,600
NORMAL MOUSE PANCREAS LOOKS

467
00:18:22,600 --> 00:18:24,560
ABSOLUTELY NORMAL AND THE IMMUNE

468
00:18:24,560 --> 00:18:26,840
COMPONENT IS ALSO VERY NORMAL.

469
00:18:26,840 --> 00:18:29,160
SO, TO ME THIS IS AN IMPORTANT

470
00:18:29,160 --> 00:18:30,320
FINDING BECAUSE IT'S THE FIRST

471
00:18:30,320 --> 00:18:36,560
TIME THAT WE REALIZE THAT THERE

472
00:18:36,560 --> 00:18:38,560
ARE THAT THE TRANSCRIPTIONAL

473
00:18:38,560 --> 00:18:40,120
COMMUNICATION ISN'T TRUE FOR ANY

474
00:18:40,120 --> 00:18:42,200
CELL IN THE PANCREAS AND THAT

475
00:18:42,200 --> 00:18:43,640
THERE ARE IMPORTANT NONCELL

476
00:18:43,640 --> 00:18:45,720
AUTONOMOUS EFFECTS THAT GIVE

477
00:18:45,720 --> 00:18:46,920
RISE TO THIS PLASTICITY SO IT'S

478
00:18:46,920 --> 00:18:48,560
1 OF THE FIRST TIMES THAT WE'VE

479
00:18:48,560 --> 00:18:51,280
BEEN ABLE TO ACTUALLY SEE

480
00:18:51,280 --> 00:18:53,120
COMMUNICATION BETWEEN P53

481
00:18:53,120 --> 00:18:54,400
TRANSCRIPTIONAL PROGRAM IN

482
00:18:54,400 --> 00:18:55,320
DIFFERENT CELLS.

483
00:18:55,320 --> 00:18:57,880
SO TO SUMMARIZE THESE DATA, I'M

484
00:18:57,880 --> 00:18:59,320
FASCINATED THAT P53, I MEAN IT

485
00:18:59,320 --> 00:19:00,480
IS A TRANSCRIPTION FACTOR.

486
00:19:00,480 --> 00:19:02,120
YOU EXPECT IT TO HAVE HUNDREDS

487
00:19:02,120 --> 00:19:04,400
OF GENES THAT WERE REGULATED BUT

488
00:19:04,400 --> 00:19:06,840
I DIDN'T EXPECT TO SEE SO MANY

489
00:19:06,840 --> 00:19:08,360
REG IEWLGTED IN A TISSUE

490
00:19:08,360 --> 00:19:08,720
SPECIFIC MANNER.

491
00:19:08,720 --> 00:19:11,160
SO IT'S ALSO SHOWING US THAT P53

492
00:19:11,160 --> 00:19:13,680
TRANSCRIPTION EFFECTS CELLS IN

493
00:19:13,680 --> 00:19:14,760
THE NONCELL AUTONOMOUS FASHION.

494
00:19:14,760 --> 00:19:16,880
AND SO 1 OF THE THINGS WE JUST

495
00:19:16,880 --> 00:19:18,280
INITTIAITED IS A SINGLE CELL

496
00:19:18,280 --> 00:19:22,840
ANALYSIS OF THE PANCREAS, TO

497
00:19:22,840 --> 00:19:24,920
DETERMINE WHAT CELLS ARE

498
00:19:24,920 --> 00:19:25,600
EXPRESSING WHICH TRANSCRIPTION

499
00:19:25,600 --> 00:19:27,400
FACTOR AND HOW ARE THEY

500
00:19:27,400 --> 00:19:28,080
COMMUNICATING WITH EACH OTHER.

501
00:19:28,080 --> 00:19:29,720
BUT THE MAJOR PART OF MY TALK

502
00:19:29,720 --> 00:19:31,640
TODAY IS ON MUTANT P53

503
00:19:31,640 --> 00:19:34,760
ACTIVITIES AND WHAT IS MUTANT

504
00:19:34,760 --> 00:19:37,440
P53, HOW IT MUTANT P53 CAUSING

505
00:19:37,440 --> 00:19:38,920
CANCER SO THE FIRST FEW SLIDES

506
00:19:38,920 --> 00:19:41,200
JUST BASICALLY TELL YOU HOW

507
00:19:41,200 --> 00:19:46,920
IMPORTANT P53 IS AS TUMOR

508
00:19:46,920 --> 00:19:47,200
SUPPRESSOR.

509
00:19:47,200 --> 00:19:49,640
THAL GRAPH, WE CALL THE

510
00:19:49,640 --> 00:19:50,960
MANHATTAN PLOT, WE LOOK LIKE

511
00:19:50,960 --> 00:19:53,160
THERE'S A BUNCH OF SKYSCRAPERS,

512
00:19:53,160 --> 00:19:57,480
IT WAS DEVELOPED BY A COLLEAGUE,

513
00:19:57,480 --> 00:19:59,120
ACROSS THE X-AXIS, THERE'S 125

514
00:19:59,120 --> 00:20:01,080
GENES THAT ARE MUTATE INDEED

515
00:20:01,080 --> 00:20:03,400
CANCER, AND ACROSS THE Y-AXIS

516
00:20:03,400 --> 00:20:04,720
THERE'S 36 DIFFERENT KINDS OF

517
00:20:04,720 --> 00:20:08,040
CANCERS AND I THINK THAT YOU CAN

518
00:20:08,040 --> 00:20:11,040
CLEARLY SEE IS THAT THAT ROW OF

519
00:20:11,040 --> 00:20:12,040
SKYSCRAPERS ON THE RIGHT HAND

520
00:20:12,040 --> 00:20:14,640
SIDE OF THE SLIDE IS WHERE P53

521
00:20:14,640 --> 00:20:16,760
IS LOCATED SO IT'S MUTATE INDEED

522
00:20:16,760 --> 00:20:18,640
MOST CANCERS.

523
00:20:18,640 --> 00:20:23,840
AND WHEN P53 ISN'T MUTATED YOU

524
00:20:23,840 --> 00:20:27,760
OFTEN HAVE PATHWAY OF THE OTHER

525
00:20:27,760 --> 00:20:28,560
MECHANISMS.

526
00:20:28,560 --> 00:20:30,200
THE OTHER FASCINATING ASPECT OF

527
00:20:30,200 --> 00:20:32,280
THE P53 TUMOR SUPPRESSOR IS THAT

528
00:20:32,280 --> 00:20:35,160
IT'S NOT USUALLY LOST, MISSENSED

529
00:20:35,160 --> 00:20:37,320
MUTATIONS ARE THE MOST COMMON

530
00:20:37,320 --> 00:20:38,400
TYPE OF P53 ALTERATION.

531
00:20:38,400 --> 00:20:40,040
SO THIS A LINE DRAWING OF P53

532
00:20:40,040 --> 00:20:42,040
AND THESE ARE THE KINDS OF

533
00:20:42,040 --> 00:20:43,880
MUTATIONS THAT OCCUR IN BREAST

534
00:20:43,880 --> 00:20:44,200
CANCER.

535
00:20:44,200 --> 00:20:45,440
IT'S SIMILAR FOR OTHER CANCERS

536
00:20:45,440 --> 00:20:47,160
BUT I'LL BE TALKING ABOUT BREAST

537
00:20:47,160 --> 00:20:50,320
MODELS AND SO THAT'S WHY I'M

538
00:20:50,320 --> 00:20:51,080
SHOWING THIS SLIDE.

539
00:20:51,080 --> 00:20:53,280
THE LITTLE BLACK CIRCLES ARE

540
00:20:53,280 --> 00:20:53,920
TRUNCATING MUTATIONS.

541
00:20:53,920 --> 00:20:57,320
AND THEN THE GREEN CIRCLES ARE

542
00:20:57,320 --> 00:20:58,720
ALL THE MISSENSE MUTATIONS AND

543
00:20:58,720 --> 00:21:01,240
WHEN I DID THE CALCULATIONS THIS

544
00:21:01,240 --> 00:21:02,960
IS ABOUT 60% MISSENSE MUTATIONS.

545
00:21:02,960 --> 00:21:05,360
THERE ARE SEVERAL HOT SPOTS, THE

546
00:21:05,360 --> 00:21:09,800
175, THE 248, AND THE 273 AMINO

547
00:21:09,800 --> 00:21:13,480
ACIDS ARE VERY, VERY OFTEN

548
00:21:13,480 --> 00:21:20,560
MUTATE INDEED CANCERS.

549
00:21:20,560 --> 00:21:21,440
THE RGENINE ARE IN THERE, THEY

550
00:21:21,440 --> 00:21:22,720
MOST OCCUR IN THE RED DOMAIN

551
00:21:22,720 --> 00:21:24,760
WHICH IS THE DNA BINDING DOMAIN

552
00:21:24,760 --> 00:21:29,840
AND THE 248 AND THE 273 ARE

553
00:21:29,840 --> 00:21:31,560
RGENINEs THAT DIRECTLY BIND

554
00:21:31,560 --> 00:21:33,560
DNA SO THOSE DISRUPT THE ABILITY

555
00:21:33,560 --> 00:21:36,960
OF THE PROTEIN TO BIND DNA.

556
00:21:36,960 --> 00:21:37,640
THE TRANSCRIPTIONAL ACTIVATION

557
00:21:37,640 --> 00:21:38,960
IS AT THE TERMINUS OF THE

558
00:21:38,960 --> 00:21:39,920
PROTEIN AND IT'S INTACT IN ALL

559
00:21:39,920 --> 00:21:42,280
OF THESE IN A SENSE MUTATIONS.

560
00:21:42,280 --> 00:21:46,240
AND THEN THE CAR BOXY TERMINUS

561
00:21:46,240 --> 00:21:47,880
CONTAINS THE TETRAMERDOMAIN

562
00:21:47,880 --> 00:21:50,080
BECAUSE P53 FUNCTIONS AS A

563
00:21:50,080 --> 00:21:51,560
TETRAMERAND IN THAT DOMAIN YOU

564
00:21:51,560 --> 00:21:54,720
ALSO SEE MISSENSED MUTATIONS

565
00:21:54,720 --> 00:21:55,960
THAT DISRUPT TETRAMERRIZATION.

566
00:21:55,960 --> 00:21:58,160
SO THERE'S SEVERAL FEATURES OF

567
00:21:58,160 --> 00:21:59,120
THESE P53 MISSENSE MUTATIONS

568
00:21:59,120 --> 00:22:01,120
THAT I WANT TO YOU KEEP IN MIND

569
00:22:01,120 --> 00:22:02,960
AS I DISCUSS OUR DATA TODAY.

570
00:22:02,960 --> 00:22:03,880
LOSS OF FUNCTION.

571
00:22:03,880 --> 00:22:05,320
THESE ARE TRUE LOSS OF FUNCTION

572
00:22:05,320 --> 00:22:09,720
ALLELES, THEY DO NOT EXPRESS A

573
00:22:09,720 --> 00:22:12,560
PROTEIN THAT CAN BIND DNA AND

574
00:22:12,560 --> 00:22:13,200
ACTIVATE TRANSCRIPTION.

575
00:22:13,200 --> 00:22:15,200
THEY ALSO HAVE DOMINANT NEGATIVE

576
00:22:15,200 --> 00:22:17,040
AND INHIBITORS EFFECTS ON

577
00:22:17,040 --> 00:22:17,280
TISSUES.

578
00:22:17,280 --> 00:22:21,560
SO BECAUSE IN MOST OF THESE D NA

579
00:22:21,560 --> 00:22:22,960
BINDING MUTANTS, THE

580
00:22:22,960 --> 00:22:23,960
TETRAMERRIZATION DOMAIN IS

581
00:22:23,960 --> 00:22:28,360
INTACT, WHAT YOU HAD IS A MIXED

582
00:22:28,360 --> 00:22:29,880
TETRAMERS THAT HAVE TILED TYPE

583
00:22:29,880 --> 00:22:31,720
AND MUTANT PROTEINS SO THAT

584
00:22:31,720 --> 00:22:32,720
MUTANT PROTEIN BASICALLY

585
00:22:32,720 --> 00:22:33,960
INHIBITS THE ACTIVITY OF THAT

586
00:22:33,960 --> 00:22:34,840
WILD-TYPE PROTEIN AND I WILL

587
00:22:34,840 --> 00:22:38,560
SHOW YOU A VERY CLEAR EXAMPLE OF

588
00:22:38,560 --> 00:22:39,080
THAT.

589
00:22:39,080 --> 00:22:40,440
BUT THEN THE OTHER FEATURE

590
00:22:40,440 --> 00:22:41,560
THAT'S OFTEN DISCUSSED IN THE

591
00:22:41,560 --> 00:22:42,600
LITERATURE AND I THINK IT'S

592
00:22:42,600 --> 00:22:44,480
REAL, I WAS ABLE TO CONVINCE

593
00:22:44,480 --> 00:22:45,560
GLEN TODAY IS WAS LEGAL IS WHAT

594
00:22:45,560 --> 00:22:47,600
WE CALL A GAIN OF FUNCTION.

595
00:22:47,600 --> 00:22:49,680
BASICALLY WHAT THIS MEAN SYSTEM

596
00:22:49,680 --> 00:22:51,800
THAT THE MUTANT P53S HAVE

597
00:22:51,800 --> 00:22:53,040
ADDITIONAL ACTIVITIES THAT

598
00:22:53,040 --> 00:22:54,760
CONTRIBUTE TO THEIR TUMOR GROWTH

599
00:22:54,760 --> 00:22:56,520
AND OFTEN TIMES THESE ACTIVITIES

600
00:22:56,520 --> 00:22:59,280
ARE DUE TO INTERACTIONS WITH

601
00:22:59,280 --> 00:23:02,680
OTHER DNA BINDING PROTEINS AND

602
00:23:02,680 --> 00:23:04,520
BASICALLY YOU UNLEASH AA

603
00:23:04,520 --> 00:23:05,800
TRANSCRIPTIONAL PROGRAM THAT'S

604
00:23:05,800 --> 00:23:08,840
NOT A P53 DEPENDENT

605
00:23:08,840 --> 00:23:10,400
TRANSCRIPTIONAL PROGRAM.

606
00:23:10,400 --> 00:23:13,200
SO THE FIRST FEW SLIDES ARE GERM

607
00:23:13,200 --> 00:23:15,440
LINE MODELS WITH THESE P53

608
00:23:15,440 --> 00:23:17,320
MISSENSED MUTATIONS BECAUSE WE

609
00:23:17,320 --> 00:23:20,200
REALLY--THE FIRST, THE R1728S

610
00:23:20,200 --> 00:23:23,240
ARE EQUIVALENT TO THE 175 AND

611
00:23:23,240 --> 00:23:24,560
THAT'S THAT CONTACT MUTANT AND

612
00:23:24,560 --> 00:23:26,000
THEN THERE'S A STRUCTURAL MUTANT

613
00:23:26,000 --> 00:23:30,160
AND THEN WE HAVE THE 245 AND THE

614
00:23:30,160 --> 00:23:31,000
270 MUTANTS, AND WE REALLY

615
00:23:31,000 --> 00:23:32,960
WANTED TO ASK A VERY SIMPLE

616
00:23:32,960 --> 00:23:37,640
QUESTION, ARE ALL MUTANT PP53S

617
00:23:37,640 --> 00:23:37,960
THE SAME?

618
00:23:37,960 --> 00:23:40,080
SO THIS IS A MODEL IN WHICH WE

619
00:23:40,080 --> 00:23:41,800
CAM PAIR THE 3 DIFFERENT ALLELES

620
00:23:41,800 --> 00:23:43,840
IN THE SAME BACKGROUND, C57

621
00:23:43,840 --> 00:23:48,320
BLACK 6 AND WE ASKED DO THESE

622
00:23:48,320 --> 00:23:50,320
THEY ALL GET TUMORS BUT DO THEY

623
00:23:50,320 --> 00:23:51,680
GET THE SAME KIND?

624
00:23:51,680 --> 00:23:52,800
SO THE FIRST EXPERIMENT WE DID

625
00:23:52,800 --> 00:23:56,800
IS ACTUALLY LOOK TO SEE WHETHER

626
00:23:56,800 --> 00:23:59,520
THE WILD-TYPE PROTEIN ACTIVITY

627
00:23:59,520 --> 00:24:01,240
VARIED DEPENDING ON THE PRESENCE

628
00:24:01,240 --> 00:24:03,000
OF THE MUTANT P53.

629
00:24:03,000 --> 00:24:06,560
SO THIS IS AN EXPERIMENT RAW

630
00:24:06,560 --> 00:24:08,000
DATA IN VIVO WE'VE TAKEN A WHOLE

631
00:24:08,000 --> 00:24:10,040
NEW MOUSE, WITH THE DIFFERENT

632
00:24:10,040 --> 00:24:10,720
GENOTYPES, RADIATED THE MOUSE

633
00:24:10,720 --> 00:24:14,760
AND LOOKED AT THE ABILITY OF

634
00:24:14,760 --> 00:24:17,080
THAT P53 TO ACTIVATE

635
00:24:17,080 --> 00:24:17,600
TRANSCRIPTION.

636
00:24:17,600 --> 00:24:21,520
SO THE FIRST GRAY BAR IS THE

637
00:24:21,520 --> 00:24:23,440
ERADIATED TISSUE AND THIS IS

638
00:24:23,440 --> 00:24:25,040
JUST PROBABLY THE THYMUS AND

639
00:24:25,040 --> 00:24:30,400
THAT SHOWS THAT FOR EXAMPLE HERE

640
00:24:30,400 --> 00:24:32,440
IN THE BAX, P53 TARGETING SYSTEM

641
00:24:32,440 --> 00:24:33,720
SET ACTIVATED 8 FOLD ABOVE

642
00:24:33,720 --> 00:24:36,360
BACKGROUND AND THEN YOU SEE THE

643
00:24:36,360 --> 00:24:39,280
CONTROLS WHICH IS THE WHITE BAR

644
00:24:39,280 --> 00:24:40,280
IS THE HETEROZYGOUS MOUSE AND

645
00:24:40,280 --> 00:24:43,000
THEN THE 3 DIFFERENT MUTANTS FOR

646
00:24:43,000 --> 00:24:44,640
ANALYZED SUBSEQUENTLY TO THAT.

647
00:24:44,640 --> 00:24:46,720
AND I THINK AS YOU LOOK ACROSS

648
00:24:46,720 --> 00:24:49,160
THE ENTIRE SLIDE, YOU CAN SEE

649
00:24:49,160 --> 00:24:52,080
THAT THOSE BARS WITH THE MUTANT

650
00:24:52,080 --> 00:24:55,160
P53, AN LOG TYPE ALLELE ARE

651
00:24:55,160 --> 00:24:57,760
ALWAYS LOWER THAN THE

652
00:24:57,760 --> 00:24:58,720
HETEROZYGOUS WHITE BAR AND WHAT

653
00:24:58,720 --> 00:25:01,160
THAT SAYS IS THAT IN RESPONSE TO

654
00:25:01,160 --> 00:25:03,480
DNA DAMAGE, THAT MUTANT PPEA IS

655
00:25:03,480 --> 00:25:05,240
ACTUALLY DAMPENING THE EFFECTS

656
00:25:05,240 --> 00:25:06,480
OF THAT WILD-TYPE P53.

657
00:25:06,480 --> 00:25:08,720
SO THIS CAN TRIBUTE TO AN

658
00:25:08,720 --> 00:25:09,520
INCREASED TUMOR PHENOTYPE

659
00:25:09,520 --> 00:25:12,280
BECAUSE YOU'VE GOT LESS

660
00:25:12,280 --> 00:25:12,880
WILD-TYPE P53.

661
00:25:12,880 --> 00:25:14,880
AND IN FACT, WHEN WE LOOK AT THE

662
00:25:14,880 --> 00:25:16,800
SURVIVAL OF THESE ANIMALS, THESE

663
00:25:16,800 --> 00:25:18,000
ANIMALS GET TUMORS, I WILL TELL

664
00:25:18,000 --> 00:25:20,720
YOU WHICH KIND IN A MINUTE, BUT,

665
00:25:20,720 --> 00:25:23,200
YOU CAN CLEARLY SEE ON THE LEFT

666
00:25:23,200 --> 00:25:26,160
WHEN WE LOOK AT A LITTLE OVER A

667
00:25:26,160 --> 00:25:29,120
YEAR, THE SAMPLES THAT HAVE THE

668
00:25:29,120 --> 00:25:32,400
BLUE AND THE BLACK, THE 245 AND

669
00:25:32,400 --> 00:25:35,080
THE 270, THESE ARE THE CONTACT

670
00:25:35,080 --> 00:25:37,280
MUTANTS, THEY GET CANCERS

671
00:25:37,280 --> 00:25:38,640
EARLIER THAN THE--THAN THE

672
00:25:38,640 --> 00:25:40,200
HETEROZYGOUS MOUSE AND THE 172

673
00:25:40,200 --> 00:25:42,240
MUTATIONS, SO THERE IS A

674
00:25:42,240 --> 00:25:44,680
DIFFERENT IN TERMS OF TUMOR

675
00:25:44,680 --> 00:25:46,080
ONSETS IN VIVO.

676
00:25:46,080 --> 00:25:47,920
AND IN ADDITION THE KINDS OF

677
00:25:47,920 --> 00:25:49,320
TUMORS THEY GET ARE ALL VERY

678
00:25:49,320 --> 00:25:52,120
SIMILAR AND THEY GET LYMPHOMAS,

679
00:25:52,120 --> 00:25:53,520
SARCOMA AND THEY GET CARCINOMAS,

680
00:25:53,520 --> 00:25:55,760
REMEMBER WE'RE IN THE SAME

681
00:25:55,760 --> 00:25:58,240
GENETIC BACKGROUND BUT THE OTHER

682
00:25:58,240 --> 00:26:00,560
MAJOR PHENOTYPE THAT WE SEE WITH

683
00:26:00,560 --> 00:26:03,800
THE MUTANTS, ALL 3 MUTANTS IS A

684
00:26:03,800 --> 00:26:05,600
METASTATIC PHENOTYPE AND TYLER

685
00:26:05,600 --> 00:26:08,000
JAX'S LAB AND MY DESCRIBED THIS

686
00:26:08,000 --> 00:26:09,520
AS THE MUTANT P53 GAIN OF

687
00:26:09,520 --> 00:26:12,280
FUNCTION PHENOTYPE YEARS AGO.

688
00:26:12,280 --> 00:26:14,320
SO, THE DIFFERENT MUTANTS, I

689
00:26:14,320 --> 00:26:15,520
THINK I SUMMARIZED, OKAY THIS IS

690
00:26:15,520 --> 00:26:18,480
1 MORE SLIDE FROM THESE DATA IS

691
00:26:18,480 --> 00:26:20,680
1 OF THE OTHER CONCEPTS, I WANT

692
00:26:20,680 --> 00:26:24,320
YOU TO TAKE HOME FROM THIS, TALK

693
00:26:24,320 --> 00:26:26,640
IS THAT MUSEUM TABT PROTEIN HAS

694
00:26:26,640 --> 00:26:28,280
TO BE PRESENT IN LIE LEVELS IN

695
00:26:28,280 --> 00:26:29,840
ORDER TO DAMPEN THE WILD-TYPE

696
00:26:29,840 --> 00:26:31,000
P53 AND IN ORDER FOR IT TO

697
00:26:31,000 --> 00:26:32,960
CONTRIBUTE TO THE GAIN OF

698
00:26:32,960 --> 00:26:34,280
FUNCTION OF THE METASTATIC

699
00:26:34,280 --> 00:26:34,760
PHENOTYPE.

700
00:26:34,760 --> 00:26:36,040
SO WHAT WE'VE DONE ON THIS

701
00:26:36,040 --> 00:26:37,120
EXPERIMENTOT RIGHT IS WE

702
00:26:37,120 --> 00:26:38,640
COMPARED ANIMALS THAT ARE

703
00:26:38,640 --> 00:26:40,920
HOMOZYGOUS FOR THESE DIFFERENT

704
00:26:40,920 --> 00:26:43,120
ALLELES AND WE BEND THEM INTO

705
00:26:43,120 --> 00:26:44,280
WHETHER THEY EXPRESS LESS THAN

706
00:26:44,280 --> 00:26:47,040
40% OF THE STABLE MUTANT P53 ON

707
00:26:47,040 --> 00:26:48,800
THE LEFT OR VERY HIGH LEVELS OF

708
00:26:48,800 --> 00:26:49,720
P53 ON THE RIGHT.

709
00:26:49,720 --> 00:26:53,800
YOU CAN SEE A TISSUE STAINED

710
00:26:53,800 --> 00:26:54,960
WITH P53 BY IMMUNOHISTOCHEMISTRY

711
00:26:54,960 --> 00:26:58,400
AND ON 91% OF THOSE CELLS HAVE

712
00:26:58,400 --> 00:27:00,200
VERY STABLE MUTANT P53 AND WHEN

713
00:27:00,200 --> 00:27:02,920
WE BEND THESE IN 2 CATEGORIES

714
00:27:02,920 --> 00:27:04,840
LESS THAN 54% STABLE IN P53

715
00:27:04,840 --> 00:27:07,200
CELLS OR GREATER THAN 60% IN

716
00:27:07,200 --> 00:27:09,240
SALE CELLS THERE'S A BIG

717
00:27:09,240 --> 00:27:11,000
DIFFERENCE IN SURVIVAL AS WELL

718
00:27:11,000 --> 00:27:12,600
AND THE SAMPLES THAT HAVE STABLE

719
00:27:12,600 --> 00:27:14,160
P53 IN RED, THOSE MICE ARE

720
00:27:14,160 --> 00:27:16,120
GETTING CANCER SOONER THAN THE

721
00:27:16,120 --> 00:27:17,840
OTHER ANIMALS THAT HAVE LESS

722
00:27:17,840 --> 00:27:19,680
STABLE P53.

723
00:27:19,680 --> 00:27:21,480
SO THIS IS 1 OF NUMEROUS PIECES

724
00:27:21,480 --> 00:27:23,000
OF DATA THAT SUGGEST THAT

725
00:27:23,000 --> 00:27:26,640
STABILITY IS CONTRIBUTING TO THE

726
00:27:26,640 --> 00:27:27,800
LETHAL EFFECTS IN THESE CELLS.

727
00:27:27,800 --> 00:27:32,560
SO JUST TO SUMMARIZE THIS BIT OF

728
00:27:32,560 --> 00:27:34,040
THE TALK.

729
00:27:34,040 --> 00:27:35,840
WE'VE MADE GERM LINE MUTATIONS

730
00:27:35,840 --> 00:27:38,440
THAT MIMIC FROM THE SYNDROME AND

731
00:27:38,440 --> 00:27:41,760
ALL 3 OF THOSE WHEN STRESSED

732
00:27:41,760 --> 00:27:44,000
DAMPEN THE ACTIVITY OF WILD-TYPE

733
00:27:44,000 --> 00:27:44,200
P53.

734
00:27:44,200 --> 00:27:45,280
THE WILD-TYPE ARE MORE

735
00:27:45,280 --> 00:27:46,680
ADDRESSIVE, THEY HAVE WORSE

736
00:27:46,680 --> 00:27:48,400
OUTCOMES THAN THE 172 OF THE P53

737
00:27:48,400 --> 00:27:50,160
KNOWN ALLELE AND THAT'S THE

738
00:27:50,160 --> 00:27:51,800
STABILITY OF THE MUTANT P53

739
00:27:51,800 --> 00:28:01,280
LEADS TO THE DECREASE SURVIVAL

740
00:28:01,280 --> 00:28:01,960
OF THOSE ANIMALS.

741
00:28:01,960 --> 00:28:07,760
SO WE STUDIED THE GERM LINE AD

742
00:28:07,760 --> 00:28:08,720
NAUSEUM LOOKING AT P53 YOU

743
00:28:08,720 --> 00:28:10,000
HADITATIONS AND ALLELES BUT WE

744
00:28:10,000 --> 00:28:13,280
REALLY WANT TO STUDY A SOMATIC

745
00:28:13,280 --> 00:28:14,160
MODEL BECAUSE SOMATIC HAPPENS

746
00:28:14,160 --> 00:28:15,280
MORE OFTEN THAN GERM LINE AND

747
00:28:15,280 --> 00:28:18,040
WHAT WE REALLY WANTED TO DO IS

748
00:28:18,040 --> 00:28:18,920
DEVELOP AN ANIMAL MODEL ON THE

749
00:28:18,920 --> 00:28:20,800
RIGHT WHERE YOU CAN MAKE A

750
00:28:20,800 --> 00:28:22,560
MUTANT P53 ONLY IN A FEW CELLS

751
00:28:22,560 --> 00:28:24,840
SURROUNDED BY A WILD-TYPE

752
00:28:24,840 --> 00:28:27,520
COMPARTMENT AND FOR US IT

753
00:28:27,520 --> 00:28:29,000
WAS--WE REALLY WANTED THIS MOUSE

754
00:28:29,000 --> 00:28:32,320
TO BE ABLE TO UNDERSTAND THE

755
00:28:32,320 --> 00:28:32,840
TUMOR MICROENVIRONMENT

756
00:28:32,840 --> 00:28:33,640
INTERACTIONS AND SO IN THE

757
00:28:33,640 --> 00:28:36,080
ANIMAL THEY WILL SHOW YOU IN A

758
00:28:36,080 --> 00:28:39,760
FEW MINUTES, THE NUTRIFILLS, THE

759
00:28:39,760 --> 00:28:42,760
CAPS, THE IMMUNE SYSTEMS, THE

760
00:28:42,760 --> 00:28:44,160
T-CELLS ARE ALL WILD-TYPE FOR

761
00:28:44,160 --> 00:28:44,400
P53.

762
00:28:44,400 --> 00:28:47,960
SO THIS IS HOW WE GENERATED

763
00:28:47,960 --> 00:28:48,560
THOSE ANIMALS.

764
00:28:48,560 --> 00:28:54,120
WE CALL IT THE P53 WM-ALLELE

765
00:28:54,120 --> 00:28:56,080
BECAUSE IT CONVERTS WILD-TYPE

766
00:28:56,080 --> 00:28:57,880
P53 TO MUTANT P53 AND THE WAI WE

767
00:28:57,880 --> 00:29:00,200
GENERATE THESE ALLELES IS

768
00:29:00,200 --> 00:29:01,680
BASICALLY UPSTREAM OF THE POINT

769
00:29:01,680 --> 00:29:04,440
MUTATION WHICH IS AN ASTERISK

770
00:29:04,440 --> 00:29:06,960
AND WE INSERTED BY WILD-TYPE

771
00:29:06,960 --> 00:29:09,280
CDNA IN THE POPULATION SEAT

772
00:29:09,280 --> 00:29:09,520
EVERSITE.

773
00:29:09,520 --> 00:29:11,480
SO NORMALLY THIS WOULD EXPRESS A

774
00:29:11,480 --> 00:29:13,000
WILD-TYPE P53 PROTEIN AND THEN

775
00:29:13,000 --> 00:29:14,880
IN A FASHION, YOU WOULD REMOVE

776
00:29:14,880 --> 00:29:16,360
THOSE WILD-TYPE SEQUENCES AND

777
00:29:16,360 --> 00:29:17,640
EXPRESS THE MUTANT PROTEIN AND

778
00:29:17,640 --> 00:29:20,200
WE DID THIS BOTH WITH THE

779
00:29:20,200 --> 00:29:21,960
STRUCTURAL MUTANT, THE 172 IN

780
00:29:21,960 --> 00:29:23,920
THE CONTACT MUTANT AND THE 245

781
00:29:23,920 --> 00:29:25,760
WHICH IS THE MOST AGGRESSIVE

782
00:29:25,760 --> 00:29:28,560
ALLELE WE HAVE IN THE LAB.

783
00:29:28,560 --> 00:29:31,280
SO THIS IS JUST 1 OF MANY SLIDES

784
00:29:31,280 --> 00:29:33,480
THAT I WON'T SHOW YOU, THAT

785
00:29:33,480 --> 00:29:34,920
BASICALLY SHOWS THAT THE ANIMALS

786
00:29:34,920 --> 00:29:37,560
THAT INHERENT 1 OF THESE

787
00:29:37,560 --> 00:29:39,640
ALLELES, EITHER THE 172 IN BLUE,

788
00:29:39,640 --> 00:29:43,080
OR THE 245 IN RED HAVE NO

789
00:29:43,080 --> 00:29:45,040
DIFFERENCES TO A NORMAL MOUSE.

790
00:29:45,040 --> 00:29:47,320
SO THESE ARE NOT TUMOR PRO

791
00:29:47,320 --> 00:29:49,520
MODELS THAT SUGGEST THAT YOU'RE

792
00:29:49,520 --> 00:29:51,800
REALLY MAKING A WILD-TYPE P53 IN

793
00:29:51,800 --> 00:29:53,920
THESE MICE.

794
00:29:53,920 --> 00:29:55,160
OKAY, SO THE FIRST EXPERIMENT I

795
00:29:55,160 --> 00:30:00,160
WILL DESCRIBE IS 1 IN WHICH

796
00:30:00,160 --> 00:30:01,120
WE'VE DONE--A BREAST MODEL AND

797
00:30:01,120 --> 00:30:04,880
WHAT WE'VE DONE IS WE'VE

798
00:30:04,880 --> 00:30:07,360
INJECTED AN ADENO VIRUS IN THE

799
00:30:07,360 --> 00:30:10,440
CREE INTO THE DUCT OF THE MEMORY

800
00:30:10,440 --> 00:30:10,680
GLAND.

801
00:30:10,680 --> 00:30:14,400
AND IF WE USE LOW TITER, WE CAN

802
00:30:14,400 --> 00:30:15,800
GET TD ON THE RED CELLS AND ON

803
00:30:15,800 --> 00:30:18,600
THE RIGHT IF WE USE A HIGHER

804
00:30:18,600 --> 00:30:19,680
TITER, WE GENERATE A WHOLE BUNCH

805
00:30:19,680 --> 00:30:26,000
OF CELLS THAT ARE POSITIVE FOR

806
00:30:26,000 --> 00:30:27,400
CREE AND TDTMTO, AND THE OTHER

807
00:30:27,400 --> 00:30:29,160
POINT I WANT TO MAKE WITH THE

808
00:30:29,160 --> 00:30:31,040
SLIDE IS WE INJECT ALWAYS

809
00:30:31,040 --> 00:30:32,800
FEMALES WHEN THEY'RE 10-12 WEEKS

810
00:30:32,800 --> 00:30:37,000
OLD AND WE USE A BACKGROUND THAT

811
00:30:37,000 --> 00:30:41,120
IS 50% C57 AND 50% BALBC, AND

812
00:30:41,120 --> 00:30:45,040
REASON WE USE THE BALB C MICE IS

813
00:30:45,040 --> 00:30:46,400
THAT THEY'RE SUSCEPTIBLE TO

814
00:30:46,400 --> 00:30:48,600
BREAST CANCER WHEREAS THE C57

815
00:30:48,600 --> 00:30:51,200
STRAIN WE'VE BEEN WORKING WITH

816
00:30:51,200 --> 00:30:53,920
ALL ALONG, RARELY GETS BREAST

817
00:30:53,920 --> 00:30:59,520
CANCER SO SOMETHING ABOUT THIS 1

818
00:30:59,520 --> 00:31:00,600
THAT YIELDS TO BREAST CANCER.

819
00:31:00,600 --> 00:31:01,880
SO THESE ARE THE DAILY BASIS

820
00:31:01,880 --> 00:31:02,520
THEA.

821
00:31:02,520 --> 00:31:03,560
AGAIN, THESE STUDIES SHOW THERE

822
00:31:03,560 --> 00:31:05,560
ARE DIFFERENCES BETWEEN THE

823
00:31:05,560 --> 00:31:09,000
ALLELES, SO THE 172 MICE ON THE

824
00:31:09,000 --> 00:31:10,080
LEFT FOR EXAMPLE, HETEROZYGOUS

825
00:31:10,080 --> 00:31:15,640
MICE BECAUSE WE WANTED TO MIMIC

826
00:31:15,640 --> 00:31:16,800
THE SOMATIC MUTANT P53 WITHOUT

827
00:31:16,800 --> 00:31:19,680
LOH, WE NEVER SAW ANY TUMORS.

828
00:31:19,680 --> 00:31:22,000
IF WE ERADIATE THAD MICE AND

829
00:31:22,000 --> 00:31:22,920
GENERATED RANDOM MUTATIONS WE

830
00:31:22,920 --> 00:31:24,720
BEGIN TO SEE TUMORS, BUT LOOK AT

831
00:31:24,720 --> 00:31:28,720
THE RIGHT HAND SIDE OF THAT

832
00:31:28,720 --> 00:31:30,440
GRAPH, THE 245 HETEROZYGOUS ALL

833
00:31:30,440 --> 00:31:32,960
BY ITSELF AT LOW DOSE AND AT

834
00:31:32,960 --> 00:31:37,400
HIGH DOSE, YIELDED BREAST

835
00:31:37,400 --> 00:31:37,800
CANCERS.

836
00:31:37,800 --> 00:31:40,560
ALL OF THE MICE THAT HAD MUTANT

837
00:31:40,560 --> 00:31:42,080
P53 HA METASTASIS BUT AGAIN THE

838
00:31:42,080 --> 00:31:45,040
245 WAS THE MOST METASTATIC

839
00:31:45,040 --> 00:31:48,560
LESION AND THE OTHER FASCINATIG

840
00:31:48,560 --> 00:31:52,600
ASPECT OF THE STORY IS ON THE

841
00:31:52,600 --> 00:31:56,560
RIGHT THERE, SO YOU DON'T ALWAYS

842
00:31:56,560 --> 00:31:58,320
APPROXIMATE SEE LOH WITH THESE,

843
00:31:58,320 --> 00:32:01,160
WITH THE 245 BUT YOU ALWAYS SAW

844
00:32:01,160 --> 00:32:02,000
LOH WITH THE 172.

845
00:32:02,000 --> 00:32:04,480
SO TO ME THIS SUGGESTS THAT

846
00:32:04,480 --> 00:32:05,960
THERE ARE DIFFERENCES IN THE

847
00:32:05,960 --> 00:32:08,160
ABILITYINGS OF THESE MUTANT

848
00:32:08,160 --> 00:32:09,560
PROTEINS TO DAMPEN WILE TYPE P53

849
00:32:09,560 --> 00:32:10,880
AND SOMETIMES YOU NEED TO LOSE

850
00:32:10,880 --> 00:32:11,640
THAT,A LEGAL AND SOMETIMES YOU

851
00:32:11,640 --> 00:32:12,000
DON'T.

852
00:32:12,000 --> 00:32:13,880
SO YOU NEED TO DAMPEN THE P53

853
00:32:13,880 --> 00:32:18,440
PATHWAY AND YOU PROBABLY DO IT

854
00:32:18,440 --> 00:32:20,680
WITH MULTIPLE MECHANISMS.

855
00:32:20,680 --> 00:32:22,360
THE OTHER I THINK FASCINATING

856
00:32:22,360 --> 00:32:23,320
PHENOTYPE THAT WE'RE STRUGGLING

857
00:32:23,320 --> 00:32:25,840
WITH AND TRYING TO UNDERSTAND IS

858
00:32:25,840 --> 00:32:27,760
THAT RIGHT COLUMN HERE IS SO

859
00:32:27,760 --> 00:32:29,560
THAT THE 245 MUTANT GAVE US 3

860
00:32:29,560 --> 00:32:31,880
DIFFERENT KINDS OF BREAST

861
00:32:31,880 --> 00:32:33,600
CANCER, BLACK, BAR IS TRIPLE

862
00:32:33,600 --> 00:32:37,760
NEGATIVE BREAST, AND THEN IN

863
00:32:37,760 --> 00:32:40,200
DARK BLUE IS HER TO ENRICHED AND

864
00:32:40,200 --> 00:32:42,960
LIGHT BLUE IS LIKELY LUMINAL B,

865
00:32:42,960 --> 00:32:44,440
SO TO ME THIS IS FASCINATING

866
00:32:44,440 --> 00:32:46,120
BECAUSE WE START OUT WITH THE

867
00:32:46,120 --> 00:32:49,440
SAME P53 MUTATION IN ALL OF

868
00:32:49,440 --> 00:32:51,640
THESE CELLS, AND YET TUMORS

869
00:32:51,640 --> 00:32:52,880
EVOLVE IN DIFFERENT DIRECTIONS

870
00:32:52,880 --> 00:32:54,440
AND I DON'T NEED TO TELL THIS

871
00:32:54,440 --> 00:32:57,040
AUDIENCE THAT YOU KNOW TRIPLE

872
00:32:57,040 --> 00:32:58,080
NEGATIVE BREAST CANCER IS HARD

873
00:32:58,080 --> 00:33:01,400
TO TREAT BUT HERE IN LUMINAL B

874
00:33:01,400 --> 00:33:02,360
THERE ARE HORMONE THERAPIES TO

875
00:33:02,360 --> 00:33:03,760
TREAT THOSE TUMORS SO WE

876
00:33:03,760 --> 00:33:06,640
EMBARKED ON IA BIG GENOME STUDY

877
00:33:06,640 --> 00:33:10,560
TO TRY TO UNDERSTAND THE RNA AND

878
00:33:10,560 --> 00:33:12,920
THE DNA TRANSCRIPTOMES TO SEE

879
00:33:12,920 --> 00:33:15,160
WHAT ARE THE CHANGES THAT ARE

880
00:33:15,160 --> 00:33:17,880
DRIVING THESE 3 DIFFERENT KINDS

881
00:33:17,880 --> 00:33:18,280
OF BREAST CANCERS.

882
00:33:18,280 --> 00:33:20,480
SO TO CONCLUDE THIS PART OF THE

883
00:33:20,480 --> 00:33:22,920
STUDY, MUTANT P53 MODELS DEVELOP

884
00:33:22,920 --> 00:33:23,760
METASTATIC BREAST CANCER, I MEAN

885
00:33:23,760 --> 00:33:26,040
THIS IS JUST A BEAUTIFUL MODEL

886
00:33:26,040 --> 00:33:29,360
OF WHAT HAPPENS IN HUMANS.

887
00:33:29,360 --> 00:33:31,920
THE 245 AS I JUST INDICATED,

888
00:33:31,920 --> 00:33:33,480
LEADS TO DEVELOPMENT OF 3

889
00:33:33,480 --> 00:33:34,200
DIFFERENT KINDS OF BREAST

890
00:33:34,200 --> 00:33:35,160
CANCERS AND THE DATA I DIDN'T

891
00:33:35,160 --> 00:33:36,560
HAVE TIME TO SHOW YOU BECAUSE I

892
00:33:36,560 --> 00:33:37,920
DO WANT TO GET TO THE OTHER PART

893
00:33:37,920 --> 00:33:41,920
OF MY TALK IS THAT, THE

894
00:33:41,920 --> 00:33:45,960
METASTASIS IN THIS MODEL, ARE

895
00:33:45,960 --> 00:33:47,000
SEATED VERY EARLY.

896
00:33:47,000 --> 00:33:49,080
THE INITIAL STUDY I THINK

897
00:33:49,080 --> 00:33:50,360
INCLUDED 20 MICE EXPW WE'VE NOW

898
00:33:50,360 --> 00:33:53,040
EXPANDED THAT STUDY TO ABOUT 100

899
00:33:53,040 --> 00:33:55,600
ANIMALS WITH THE 245 MUTATION

900
00:33:55,600 --> 00:33:56,720
AND WHICH ACI HAVE FIESED MOST

901
00:33:56,720 --> 00:34:00,400
OF THESE MICE AND SOME OF THEM

902
00:34:00,400 --> 00:34:01,560
WE SACRIFICED BECAUSE THEY HAD

903
00:34:01,560 --> 00:34:03,400
DIFFICULTY BREATHING AND THE

904
00:34:03,400 --> 00:34:05,640
VETERINARIAN SAID WE HAVE TO

905
00:34:05,640 --> 00:34:07,680
SACRIFICE THE MICE, SO WHEN WE

906
00:34:07,680 --> 00:34:08,680
SACRIFICE THE MICE WE HAVE A LOT

907
00:34:08,680 --> 00:34:10,640
OF LESIONS IN THE LUNG YOU BUT

908
00:34:10,640 --> 00:34:12,320
WE CAN'T FIND A PRIMARY BEST

909
00:34:12,320 --> 00:34:15,360
TUMOR SO WE THINK THIS IS JUST

910
00:34:15,360 --> 00:34:17,600
ANOTHER EXAMPLE OF EARLY

911
00:34:17,600 --> 00:34:23,280
DISSEMINATION OF THAT PRIMARY

912
00:34:23,280 --> 00:34:23,520
CANCER.

913
00:34:23,520 --> 00:34:24,720
SO THE LAST MODEL I WILL DISCUSS

914
00:34:24,720 --> 00:34:26,600
WITH YOU TODAY IS 1 IN WHICH WE

915
00:34:26,600 --> 00:34:28,560
USED THIS SYSTEM TO BASICALLY

916
00:34:28,560 --> 00:34:30,200
TURN ON THE MUTANT P53 AND THEN

917
00:34:30,200 --> 00:34:32,960
TAKE IT AWAY AND WE WANT TO ASK

918
00:34:32,960 --> 00:34:34,840
THE QUESTIONS, ARE THOSE TUMORS

919
00:34:34,840 --> 00:34:37,680
DEPENDENT ON HAVING THAT MUTANT

920
00:34:37,680 --> 00:34:38,000
P53.

921
00:34:38,000 --> 00:34:39,880
SO, WE START OUT WITH THE

922
00:34:39,880 --> 00:34:42,480
WILD-TYPE P53, THESE ARE THESE

923
00:34:42,480 --> 00:34:44,640
WM ALLELES, WE THEN IN THIS

924
00:34:44,640 --> 00:34:47,320
MODEL WE USE K14 CREE, SO WE'RE

925
00:34:47,320 --> 00:34:50,000
EXPRESSING CREE RECOMBIN ACE IN

926
00:34:50,000 --> 00:34:52,560
THE ENTIRE EPITHELIUM OF THE

927
00:34:52,560 --> 00:34:56,240
BREAST AND THE--THE SALIVARY

928
00:34:56,240 --> 00:34:58,360
GLANDS AND THE SKIN AND THEN A

929
00:34:58,360 --> 00:34:59,840
YEAR LATER WE GET BREAST CANCER

930
00:34:59,840 --> 00:35:01,760
AND WE ALSO GET HEAD AND NECK

931
00:35:01,760 --> 00:35:03,720
CANCERS BUT WE HAVEN'T STUDIED

932
00:35:03,720 --> 00:35:06,680
THOSE, SO FOR THE BREAST MODELS

933
00:35:06,680 --> 00:35:07,880
BECAUSE WE'VE ALSO TURNED ON

934
00:35:07,880 --> 00:35:10,080
CAS9 WHEN WE TURNED ON K14 CREE,

935
00:35:10,080 --> 00:35:13,520
SO NOW WE'RE ABLE TO COME IN,

936
00:35:13,520 --> 00:35:16,080
WITH A GUIDE RNA, THE ADENO

937
00:35:16,080 --> 00:35:17,520
VIRUS EXPRESSING A GUIDE RNA TO

938
00:35:17,520 --> 00:35:19,720
TAKE OUT THE MUTANT P53 AND WE

939
00:35:19,720 --> 00:35:20,800
CAN ASK WHAT HAPPENS TO THIS

940
00:35:20,800 --> 00:35:25,160
CELL WHEN YOU REMOVE THAT MUTANT

941
00:35:25,160 --> 00:35:25,680
PPEA.

942
00:35:25,680 --> 00:35:27,720
SO IMPORTANTLY, THESE MICE ONLY

943
00:35:27,720 --> 00:35:30,760
EXPRESS 1 MUTANT P53 AND 1 KNOWN

944
00:35:30,760 --> 00:35:32,520
ALLELE, SO THERE'S NO WILD-TYPE

945
00:35:32,520 --> 00:35:34,400
P53, THERE'S NO LOH, WE DID IT

946
00:35:34,400 --> 00:35:39,120
FROM THE BEGINNING, MUTANT P53S

947
00:35:39,120 --> 00:35:40,440
IS ONLY EXPRESSED IN THE SYSTEM.

948
00:35:40,440 --> 00:35:43,640
AND SOME OF THESE HAVE TMTD, SO

949
00:35:43,640 --> 00:35:46,920
WE CAN MONITOR THE PRIMARY TUMOR

950
00:35:46,920 --> 00:35:48,200
DEVELOPMENT AND THE METASTASIS.

951
00:35:48,200 --> 00:35:51,760
SO THE K14 CREE MODEL GIVES US

952
00:35:51,760 --> 00:35:52,280
TRIPLE NEGATIVE BREAST.

953
00:35:52,280 --> 00:35:56,280
ALL THE MICE ARE DEAD WITHIN

954
00:35:56,280 --> 00:35:59,000
LIKE A YEAR AND A HALF.

955
00:35:59,000 --> 00:36:02,400
THEY GET 2 KINDS OF BREAST

956
00:36:02,400 --> 00:36:04,200
CARCINOMAS 1 IS THE ADEN O

957
00:36:04,200 --> 00:36:07,360
CARCINOMA AND THE OTHER IS A

958
00:36:07,360 --> 00:36:08,840
SARCOMAATOID LIKE TUMOR AND THEY

959
00:36:08,840 --> 00:36:10,120
HAVE VERY DIFFERENT ETIOLOGIES.

960
00:36:10,120 --> 00:36:13,440
SO WHAT WE'VE DONE THEN IS

961
00:36:13,440 --> 00:36:16,280
INJECTED THESE TUMORS AFTER THEY

962
00:36:16,280 --> 00:36:18,480
DEVELOPED WITH THE ADEN O

963
00:36:18,480 --> 00:36:20,120
ASSOCIATED VIRUS EXPRESSING A

964
00:36:20,120 --> 00:36:22,720
GUIDE RNA TO THE MUTANT P53, SO

965
00:36:22,720 --> 00:36:24,920
DEPLETING THE TUMORS OF MUTANT

966
00:36:24,920 --> 00:36:27,160
P53 AND ON THE LEFT I HAVE A.

967
00:36:27,160 --> 00:36:28,240
 Q.OF THAT SHOWS THE

968
00:36:28,240 --> 00:36:29,320
CONTROLS IN A, SO THIS

969
00:36:29,320 --> 00:36:31,160
IS HA HAPPEN WHEN IS

970
00:36:31,160 --> 00:36:32,960
MUTANT P53 IS STILL

971
00:36:32,960 --> 00:36:34,120
BEING EXPRESSED AND

972
00:36:34,120 --> 00:36:35,960
YOU CAN SEE THAT THE

973
00:36:35,960 --> 00:36:38,040
TUMOR VOLUME CONTINUES

974
00:36:38,040 --> 00:36:39,880
TO INCREASE IN PURPLE,

975
00:36:39,880 --> 00:36:42,960
OR THE TUMORS THAT NOW

976
00:36:42,960 --> 00:36:43,440
HAVE DEPLETED MUTANT

977
00:36:43,440 --> 00:36:44,520
P53 AND I THINK YOU

978
00:36:44,520 --> 00:36:45,560
CAN SEE A BIG

979
00:36:45,560 --> 00:36:47,040
DIFFERENCE BETWEEN

980
00:36:47,040 --> 00:36:48,240
THOSE BARS, SO SOME OF

981
00:36:48,240 --> 00:36:50,400
THE TUMORS HAVE A

982
00:36:50,400 --> 00:36:54,080
DECREASED VOLUME AND

983
00:36:54,080 --> 00:36:54,760
SOME HAVE A LARGE

984
00:36:54,760 --> 00:36:55,720
DECREASE IN OW QUICKLY

985
00:36:55,720 --> 00:36:59,120
THAT TUMOR IS GROWING.

986
00:36:59,120 --> 00:37:01,560
ON THE RIGHT YOU SEE THE TUMOR

987
00:37:01,560 --> 00:37:06,440
VOLUME WITH TIME, AGAIN IN

988
00:37:06,440 --> 00:37:08,520
PURPLE ARE THE MICE THAT HAVE

989
00:37:08,520 --> 00:37:09,480
DEPLETED P53 AND IN GREEN ARE

990
00:37:09,480 --> 00:37:13,800
THE MICE THAT HAVE HIGH LEVELS

991
00:37:13,800 --> 00:37:15,600
OF THAT MUTANT PROTEIN.

992
00:37:15,600 --> 00:37:19,360
IN ORANGE ARE 2 INTERESTING

993
00:37:19,360 --> 00:37:22,240
SAMPLES THAT LOOK LIKE MUTANT

994
00:37:22,240 --> 00:37:24,440
P53, THEY'VE NOT--THEY'RE

995
00:37:24,440 --> 00:37:25,800
CONTINUING TO GROW.

996
00:37:25,800 --> 00:37:28,000
THIS WAS A--THESE WERE 2 SAMPLES

997
00:37:28,000 --> 00:37:32,960
FOR WHICH THE MUTANT P53 WAS NOT

998
00:37:32,960 --> 00:37:33,200
STABLE.

999
00:37:33,200 --> 00:37:36,280
SO YOU NEED TO HAVE A STABLE

1000
00:37:36,280 --> 00:37:38,760
MUTANT P53 FOR THE TUMORS TO BE

1001
00:37:38,760 --> 00:37:40,920
DEPENDENT ON THAT MUTANT P53.

1002
00:37:40,920 --> 00:37:43,880
NEXT IS JUST A SURVIVAL KUEVER

1003
00:37:43,880 --> 00:37:45,640
SO IN PURPLE ARE THE ANIMALS FOR

1004
00:37:45,640 --> 00:37:47,080
WHICH WE REMOVE THAT P53 PROTEIN

1005
00:37:47,080 --> 00:37:48,400
AND YOU CAN SEE THAT THEY'RE

1006
00:37:48,400 --> 00:37:50,400
LIVING A LOT LONGER.

1007
00:37:50,400 --> 00:37:53,240
THIS IS PROBABLY AN

1008
00:37:53,240 --> 00:37:54,440
UNDERESTIMATE OF HOW LONG THEY

1009
00:37:54,440 --> 00:37:56,200
LIVE BECAUSE SOME OF THESE

1010
00:37:56,200 --> 00:37:57,240
ANIMALS GET SECONDARY TUMORS AND

1011
00:37:57,240 --> 00:37:59,120
YOU HAVE TO SACRIFICE THEM

1012
00:37:59,120 --> 00:38:00,880
BECAUSE OF THE SECONDARY TUMOR

1013
00:38:00,880 --> 00:38:04,720
AND NOT THE PRIMARY TUMOR THAT

1014
00:38:04,720 --> 00:38:05,800
WE TREATED.

1015
00:38:05,800 --> 00:38:08,440
THE OTHER MUTATION, THE 245,

1016
00:38:08,440 --> 00:38:10,440
IT'S THE SAME STORY, WE DELETE

1017
00:38:10,440 --> 00:38:11,880
THE MUTANT P53 AND YOU CAN SEE

1018
00:38:11,880 --> 00:38:13,720
IN PURPLE THAT THESE TUMORS

1019
00:38:13,720 --> 00:38:15,760
EITHER REGRESS OR DRASTICALLY

1020
00:38:15,760 --> 00:38:18,000
SLOW DOWN AND CAN YOU SEE ON THE

1021
00:38:18,000 --> 00:38:20,320
RIGHT, AND SURVIVAL KUEVER, YOU

1022
00:38:20,320 --> 00:38:21,160
ALSO EXTEND SURVIVAL OF THESE

1023
00:38:21,160 --> 00:38:26,360
MICE WHEN YOU REMOVE THAT MUTANT

1024
00:38:26,360 --> 00:38:26,960
P53 PROTEIN.

1025
00:38:26,960 --> 00:38:28,240
AND THIS IS THE EXPERIMENT WHERE

1026
00:38:28,240 --> 00:38:30,400
I WANT TO SHOW YOU THE

1027
00:38:30,400 --> 00:38:30,840
RESPONDERS VERSUS THE

1028
00:38:30,840 --> 00:38:35,480
NONRESPONDERS SO HAVE YOU

1029
00:38:35,480 --> 00:38:38,920
THIS--THIS IS A SECTION THROUGH

1030
00:38:38,920 --> 00:38:39,840
THE TUMORS.

1031
00:38:39,840 --> 00:38:41,360
THE AAV CONTROL IS ON THE LEFT

1032
00:38:41,360 --> 00:38:43,480
AND EVERYTHING IN RED IS P53, SO

1033
00:38:43,480 --> 00:38:44,880
THE 2 RESPONDERS IN THE MIDDLE,

1034
00:38:44,880 --> 00:38:47,040
CAN YOU SEE THE HUGE HOLE THAT

1035
00:38:47,040 --> 00:38:48,560
DOESN'T HAVE ANY STAINING AND

1036
00:38:48,560 --> 00:38:50,760
THAT'S WHERE WE DEPLETED THE

1037
00:38:50,760 --> 00:38:54,720
MUTANT P53 IN BOTH OF THOSE

1038
00:38:54,720 --> 00:38:56,120
TUMORS REGRESSED, THE VOLUMES

1039
00:38:56,120 --> 00:38:57,800
REGRESSED AND ON THE RIGHT YOU

1040
00:38:57,800 --> 00:38:58,840
HAVE A NONRESPONDER SO THAT YOU

1041
00:38:58,840 --> 00:39:01,720
CAN CLEARLY SEE THERE'S NO

1042
00:39:01,720 --> 00:39:03,480
STABLE MUTANT P53 IN THIS

1043
00:39:03,480 --> 00:39:04,480
RESPONDER SO THIS RESPONDER

1044
00:39:04,480 --> 00:39:06,280
LOOKS LIKE IT HAD NO P53 TO

1045
00:39:06,280 --> 00:39:08,560
START WITH SO IT WASN'T

1046
00:39:08,560 --> 00:39:09,680
DEPENDENT ON THE MUTANT P53.

1047
00:39:09,680 --> 00:39:11,720
SO I THINK THAT THIS IS AN

1048
00:39:11,720 --> 00:39:13,160
IMPORTANT EXPERIMENT, BECAUSE I

1049
00:39:13,160 --> 00:39:14,440
THINK IN THE CLINIC WE NEED TO

1050
00:39:14,440 --> 00:39:16,520
THINK ABOUT NOT JUST THE POINT

1051
00:39:16,520 --> 00:39:17,720
MUTATION BUT WHETHER IT'S BECOME

1052
00:39:17,720 --> 00:39:18,640
A STABLE PROTEIN OR NOT.

1053
00:39:18,640 --> 00:39:22,520
SO WE MIGHT HAVE TO REFER BACK

1054
00:39:22,520 --> 00:39:24,080
TO IMMUNOHISTOCHEMISTRY TO

1055
00:39:24,080 --> 00:39:26,040
DETECT MUTANT P53 STABILITY.

1056
00:39:26,040 --> 00:39:27,880
SO THE MUTANT PPARADOXICAL

1057
00:39:27,880 --> 00:39:31,520
PROTEINS BOTH THE 172 AND THE

1058
00:39:31,520 --> 00:39:33,920
245, ARE REQUIRED FOR

1059
00:39:33,920 --> 00:39:35,240
MAINTENANCE OF THESE TUMOR

1060
00:39:35,240 --> 00:39:37,000
PHENOTYPES BECAUSE WHEN WE TAKE

1061
00:39:37,000 --> 00:39:38,720
THESE 2 PROTEINS OUT, TUMORS

1062
00:39:38,720 --> 00:39:40,960
REGRESS, AND MOST OF THESE

1063
00:39:40,960 --> 00:39:43,080
TUMORS HAVE STABILIZED THE

1064
00:39:43,080 --> 00:39:44,760
MUTANT P53, A FEW DO NOT AND I

1065
00:39:44,760 --> 00:39:45,360
THINK THAT'S ANOTHER IMPORTANT

1066
00:39:45,360 --> 00:39:46,320
QUESTION IN THE FIELD THAT WE'VE

1067
00:39:46,320 --> 00:39:49,480
NOT BEEN ABLE TO FULLY

1068
00:39:49,480 --> 00:39:51,120
UNDERSTAND WHAT IS IT IN THESE

1069
00:39:51,120 --> 00:39:52,520
TUMORS THAT IS CAUSING THE

1070
00:39:52,520 --> 00:39:55,240
STABILITY OF THAT MUTANT P53.

1071
00:39:55,240 --> 00:39:59,040
YOU KNOW, IS THERE SOME STRESS,

1072
00:39:59,040 --> 00:40:00,440
DAMAGES, INFLAMMATION, ARE THOSE

1073
00:40:00,440 --> 00:40:02,040
CONTRIBUTING TO THE INSTABILITY

1074
00:40:02,040 --> 00:40:04,520
OF P53 PROTEIN AND THEN IF THE

1075
00:40:04,520 --> 00:40:06,240
TUMORS HAVE AN UNSTABLE P53

1076
00:40:06,240 --> 00:40:07,880
THEY'RE NOT DEPENDENT ON THAT

1077
00:40:07,880 --> 00:40:11,560
MUTANT P53 FOR CONTINUED GROWTH.

1078
00:40:11,560 --> 00:40:15,040
SO NOW THAT WE KNOW THAT TAKING

1079
00:40:15,040 --> 00:40:17,000
OUTLET MUSEUM TABT P53 CAUSES

1080
00:40:17,000 --> 00:40:18,640
THESE TUMORS TO REGRESS, WE'RE

1081
00:40:18,640 --> 00:40:19,600
REALLY INTERESTED IN TRYING TO

1082
00:40:19,600 --> 00:40:21,320
UNDERSTAND HOW THAT HAPPENS, SO

1083
00:40:21,320 --> 00:40:24,360
IN THIS EXPERIMENT, WHAT WE'VE

1084
00:40:24,360 --> 00:40:27,360
DONE IS WE'VE IN PURPLE AGAIN WE

1085
00:40:27,360 --> 00:40:33,240
TREATED THESE TUMORS WITH THE

1086
00:40:33,240 --> 00:40:34,760
ASSOCIATED GUIDE RNA THAT

1087
00:40:34,760 --> 00:40:35,600
DEPLETES MUTANT P53 AND YOU CAN

1088
00:40:35,600 --> 00:40:40,040
SEE THAT THE TUMORS DON'T GROW

1089
00:40:40,040 --> 00:40:40,760
AS QUICKLY.

1090
00:40:40,760 --> 00:40:42,480
THIS IS JUST 6 DAYS, SO IT'S

1091
00:40:42,480 --> 00:40:44,480
VERY EARLY ON, BECAUSE WE WANTED

1092
00:40:44,480 --> 00:40:46,480
TO CATCH THE EARLY EVENTS TO TRY

1093
00:40:46,480 --> 00:40:48,920
TO UNDERSTAND WHAT THE TRIGGER

1094
00:40:48,920 --> 00:40:49,160
WAS.

1095
00:40:49,160 --> 00:40:51,920
SO IN THE MIDDLE PANELS WHAT WE

1096
00:40:51,920 --> 00:40:53,680
JUST DID SOME H&  E AND
LOOKED

1097
00:40:53,680 --> 00:40:55,960
AT THESE AS THESE TUMORS AND

1098
00:40:55,960 --> 00:40:57,920
WHEN WE DEPLETE MUTANT P53, WE

1099
00:40:57,920 --> 00:41:03,600
CAN SEE THESE VERY LARGE LIPID

1100
00:41:03,600 --> 00:41:04,800
LIKE STRUCTURES AND THEY'RE

1101
00:41:04,800 --> 00:41:07,000
QUANTIFIED ON THE RIGHT, SO WHEN

1102
00:41:07,000 --> 00:41:09,160
WE DEPLETE P53 WE SEE THESE

1103
00:41:09,160 --> 00:41:11,680
LARGE STRUCTURES VERY

1104
00:41:11,680 --> 00:41:14,800
REMINISCENT OF A PROCESS CALLED

1105
00:41:14,800 --> 00:41:16,640
CELL APOPTOSIS, SO WE STAINED

1106
00:41:16,640 --> 00:41:18,640
THESE WITH 4 HNE WHICH IS

1107
00:41:18,640 --> 00:41:20,280
BASICALLY A MARKER FOR LIPID

1108
00:41:20,280 --> 00:41:21,440
PEROXIDATION AND I THINK CAN YOU

1109
00:41:21,440 --> 00:41:26,160
CLEARLY SEE ALL THAT BROWN

1110
00:41:26,160 --> 00:41:28,360
STAINING IN THE MIDDLE WHEN WE

1111
00:41:28,360 --> 00:41:29,440
DEPLETE MUTANT P53 WE HAVE A

1112
00:41:29,440 --> 00:41:35,800
HIGH LEVEL OF LIPID

1113
00:41:35,800 --> 00:41:38,240
PEROXIDATION, SO THESE THAT

1114
00:41:38,240 --> 00:41:39,560
SUGGEST TO US THAT MAYBE WE'RE

1115
00:41:39,560 --> 00:41:41,040
DEALING WITH A CELL DETH

1116
00:41:41,040 --> 00:41:43,200
MECHANISM SO NEXT EXPERIMENT WAS

1117
00:41:43,200 --> 00:41:45,120
SIMPLY TO ISOLATE THOSE TUMOR

1118
00:41:45,120 --> 00:41:47,000
CELLS AWAY FROM ALL THE REST OF

1119
00:41:47,000 --> 00:41:48,040
THE MICROENVIRONMENT TO SEE WHAT

1120
00:41:48,040 --> 00:41:49,800
WAS ACTUALLY HAPPENING IN THOSE

1121
00:41:49,800 --> 00:41:53,640
CELLS SO WE DID SOME SINGLE CELL

1122
00:41:53,640 --> 00:41:56,120
ANALYSIS, WE'VE IDENTIFIED 3

1123
00:41:56,120 --> 00:42:02,680
CLUSTERS, 1, 11 AND 14 THAT WERE

1124
00:42:02,680 --> 00:42:04,120
THE TUMOR CELLS AND YOU CAN SEE

1125
00:42:04,120 --> 00:42:06,160
IN THE CLOAL THESE CLUSTERS ARE

1126
00:42:06,160 --> 00:42:08,160
PRESENT AT HIGH LEVELS AND WHEN

1127
00:42:08,160 --> 00:42:09,720
WE DEPLETE THESE, THESE CLUSTERS

1128
00:42:09,720 --> 00:42:11,080
ARE LOST, SO THESE ARE THE

1129
00:42:11,080 --> 00:42:18,000
CLUSTERS THAT ARE THE TUMOR

1130
00:42:18,000 --> 00:42:19,640
CELLS THAT EXPRESS THE MUTANT

1131
00:42:19,640 --> 00:42:21,080
P53 PROTEIN SO OF COURSE YOU DO

1132
00:42:21,080 --> 00:42:24,640
THESE ANALYSIS AND YOU LOOK AT

1133
00:42:24,640 --> 00:42:28,200
THE TRANSCRIPTOME AND IN THESE

1134
00:42:28,200 --> 00:42:29,960
SAMPLES WITH THE DEPLETION, WE

1135
00:42:29,960 --> 00:42:32,680
SAW UPREGULATION OF REACTIVE

1136
00:42:32,680 --> 00:42:34,720
OXYGEN SPECIES SO AGAIN, THIS IS

1137
00:42:34,720 --> 00:42:39,760
A MECHANISM THAT INCREASED R. O.

1138
00:42:39,760 --> 00:42:40,280
S. LEADS TO THERAPY AND

1139
00:42:40,280 --> 00:42:41,760
OPERATING GLOBALLY TO THETIC

1140
00:42:41,760 --> 00:42:48,560
RESPONSE, SO WE'RE BEGINNING TO

1141
00:42:48,560 --> 00:42:49,240
THING THAT THERRAPITOSEIS.

1142
00:42:49,240 --> 00:42:50,040
SO WHAT IS IT?

1143
00:42:50,040 --> 00:42:55,960
WE'RE LEARNING A LOT ABOUT

1144
00:42:55,960 --> 00:42:56,440
THERRAPOTIIS.

1145
00:42:56,440 --> 00:42:59,040
IT'S 2 OF THE SPECIES CAUSES

1146
00:42:59,040 --> 00:43:01,000
LIPID PEROXIDATION, IT'S AN IRON

1147
00:43:01,000 --> 00:43:02,560
DEPENDENT PROCESS AND BASICALLY

1148
00:43:02,560 --> 00:43:04,040
IT KILLS THE CELL.

1149
00:43:04,040 --> 00:43:05,800
SO, ON THE LEFT CAN YOU SEE A

1150
00:43:05,800 --> 00:43:10,920
LIPID MOLECULE WITH A LOT OF

1151
00:43:10,920 --> 00:43:15,640
HYDROXYL RADICALS AND THAT WILL

1152
00:43:15,640 --> 00:43:18,240
LEAD TO FERROPITOSEIS, AND

1153
00:43:18,240 --> 00:43:20,200
THERE'S A GLUTEO THIGH OWN

1154
00:43:20,200 --> 00:43:22,000
TRANSFER ACE WHICH REMOVES THESE

1155
00:43:22,000 --> 00:43:23,160
RADICALS AND ALLOWS THE CELL TO

1156
00:43:23,160 --> 00:43:25,920
SURVIVE SO IT'S A VERY CRITICAL

1157
00:43:25,920 --> 00:43:29,440
PROTEIN IN THE CELL THAT

1158
00:43:29,440 --> 00:43:31,280
MAINTAINS THE PEROXIDATION AT A

1159
00:43:31,280 --> 00:43:32,280
LEVEL THAT'S TOLERATED.

1160
00:43:32,280 --> 00:43:33,120
SO THE NEXT EXPERIMENT I WILL

1161
00:43:33,120 --> 00:43:38,480
SHOW YOU IS 1 IN WHICH WE'VE

1162
00:43:38,480 --> 00:43:47,360
INHIBITED GBX 4, SO BASICALLY WE

1163
00:43:47,360 --> 00:43:48,560
INDUCED FERROPITOSEIS IN THESE

1164
00:43:48,560 --> 00:43:49,360
CELLS.

1165
00:43:49,360 --> 00:43:50,560
SO IT'S AN INTERESTING

1166
00:43:50,560 --> 00:43:51,000
EXPERIMENT.

1167
00:43:51,000 --> 00:43:53,520
WE HAVE IN GREEN CELLS EITHER

1168
00:43:53,520 --> 00:43:56,040
EXPRESS THE MUTANT 172, OR THE

1169
00:43:56,040 --> 00:43:58,160
MUTANT 245 AND THEN YOU USE

1170
00:43:58,160 --> 00:44:00,560
CRSPR TO REMOVE IT FROM THESE

1171
00:44:00,560 --> 00:44:02,080
CELLS IN MY POST DOCKER CAME TO

1172
00:44:02,080 --> 00:44:05,200
ME AND SAID, I HAVE CELLS WITH

1173
00:44:05,200 --> 00:44:06,920
DELETION P53 AND SAID IN VIVO

1174
00:44:06,920 --> 00:44:08,480
THOSE CELLS ARE DYING HOW DID

1175
00:44:08,480 --> 00:44:09,760
YOU GET THOSE CELLS TO LIVE AND

1176
00:44:09,760 --> 00:44:13,600
IT TURNS OUT THEY'RE VERY

1177
00:44:13,600 --> 00:44:15,040
FRAGILE, WE HAVE TO CHANGE THE

1178
00:44:15,040 --> 00:44:17,120
MEDIA EVERY DAY BECAUSE IT TURNS

1179
00:44:17,120 --> 00:44:19,480
VERY YELLOW.

1180
00:44:19,480 --> 00:44:22,160
AND SO, THE TOLERATED LOSS OF

1181
00:44:22,160 --> 00:44:25,560
P53 IN CULTURE BUT THEY'RE VERY

1182
00:44:25,560 --> 00:44:29,320
FRAGILE AND WHEN WE ADD RSL 3,

1183
00:44:29,320 --> 00:44:31,560
WE CAN SEE THE CELLS HAVE NO

1184
00:44:31,560 --> 00:44:34,640
P53, DELETION ARE VERY SENSITIVE

1185
00:44:34,640 --> 00:44:36,440
TO FERROPITOSEIS AND I THINK THE

1186
00:44:36,440 --> 00:44:37,520
DRAMATIC SAMPLE ON THE FAR RIGHT

1187
00:44:37,520 --> 00:44:41,200
IS 1 IN WHICH, THE CELL THAT HAS

1188
00:44:41,200 --> 00:44:43,400
THE MUTANT P53 IS RESISTANT AND

1189
00:44:43,400 --> 00:44:46,920
THE CELL THAT HAS LOST P53 IS

1190
00:44:46,920 --> 00:44:52,040
JUST--IT DIES VERY QUICKLY.

1191
00:44:52,040 --> 00:44:55,760
SO WE'VE ALSO INHIBITED

1192
00:44:55,760 --> 00:44:56,760
FERROPITOSEIS IN VIVO.

1193
00:44:56,760 --> 00:44:58,400
SOPHISTICATEDY THIS IS AN

1194
00:44:58,400 --> 00:44:59,800
EXPERIMENT THAT I'VE PREVIOUSLY

1195
00:44:59,800 --> 00:45:02,000
SHOWN WHERE YOU CAN SEE IF YOU

1196
00:45:02,000 --> 00:45:04,000
DEPLETE MUTANT P53 IN PURPLE

1197
00:45:04,000 --> 00:45:07,280
THESE ANIMALS LIVE LONGER SO

1198
00:45:07,280 --> 00:45:09,400
WHAT WE'VE DONE IS WE'VE

1199
00:45:09,400 --> 00:45:15,680
DEPLETED MUTANT P53 AND THEN WE

1200
00:45:15,680 --> 00:45:16,840
ADDED LACK ROUGH ATOM STATIN

1201
00:45:16,840 --> 00:45:22,680
WHICH IS AN INHIBITOR OF

1202
00:45:22,680 --> 00:45:23,840
FERROPITOSEIS, THE DATA ARE

1203
00:45:23,840 --> 00:45:25,760
QUANTIFIED IN THE MIDDLE, SO

1204
00:45:25,760 --> 00:45:28,080
WHEN WE PREVENT FERROPITOSEIS

1205
00:45:28,080 --> 00:45:30,320
FROM HAPPENING, THOSE ANIMALS

1206
00:45:30,320 --> 00:45:31,480
ARE DYING SOONER AND THAT DATA

1207
00:45:31,480 --> 00:45:34,480
THEN IS SHOWN ON THE RIGHT.

1208
00:45:34,480 --> 00:45:37,680
THE MUTANT P53 DEPLETED TUMORS

1209
00:45:37,680 --> 00:45:40,080
CONTINUE TO--THEY LIVE LONGER,

1210
00:45:40,080 --> 00:45:43,040
THE MICE WITH MUTANT P53

1211
00:45:43,040 --> 00:45:47,480
DEPLETED LIVE LONGER BUT IF YOU

1212
00:45:47,480 --> 00:45:48,680
INDUCE FERROPITOSEIS, THEY'RE

1213
00:45:48,680 --> 00:45:48,880
DYING.

1214
00:45:48,880 --> 00:45:49,840
AND I THINK THIS IS VERY CLEAR.

1215
00:45:49,840 --> 00:45:54,200
EACH 1 OF THESE IS A--IT'S NOT A

1216
00:45:54,200 --> 00:45:55,920
SIN GENETIC MOUSE, IT'S A REAL

1217
00:45:55,920 --> 00:45:57,120
MOUSE THAT GEPPED A TUMOR AND

1218
00:45:57,120 --> 00:45:59,720
THEN ONCE YOU SAW THE TUMOR WE

1219
00:45:59,720 --> 00:46:02,480
TREATED IT WITH THE LIPO STATIN.

1220
00:46:02,480 --> 00:46:05,600
SO THE HYPOTHESIS THAT WE'RE

1221
00:46:05,600 --> 00:46:08,240
WORKING WITH IS THAT P53OXIDATES

1222
00:46:08,240 --> 00:46:11,320
THIS STRESS AND PROTECTS ITS

1223
00:46:11,320 --> 00:46:13,680
FROM FERROPITOSEIS, WE REMOVE

1224
00:46:13,680 --> 00:46:15,040
THE P53 MUTANT IN THE CELL

1225
00:46:15,040 --> 00:46:18,040
TISSUE, THE CELLS ARE DYING BY

1226
00:46:18,040 --> 00:46:18,560
FERROPITOSEIS.

1227
00:46:18,560 --> 00:46:20,840
SO HOW ON TO TEST THIS.

1228
00:46:20,840 --> 00:46:22,720
SO HOW THE FIRST THING WE DID WE

1229
00:46:22,720 --> 00:46:26,120
WENT BACK TO OUR SINGLE CELL RNA

1230
00:46:26,120 --> 00:46:29,080
SEQ DAT IS AND WE LOOKED AT THE

1231
00:46:29,080 --> 00:46:30,520
REACTIVEOX GENERATED SPECIES,

1232
00:46:30,520 --> 00:46:32,560
THE GENES THAT ARE RESPONSIBLE

1233
00:46:32,560 --> 00:46:36,720
FOR ROS, AND THERE ARE 2 GENES

1234
00:46:36,720 --> 00:46:42,920
THAT STUCK OUT, PIDRX 6 AND MGST

1235
00:46:42,920 --> 00:46:45,600
3, AND PIDR6 A GLUTATHIONE

1236
00:46:45,600 --> 00:46:48,120
DEPENDENT SO IT'S GOING TO WORK

1237
00:46:48,120 --> 00:46:49,840
SIMILARLY TO GPX 4 AND REMOVE

1238
00:46:49,840 --> 00:46:52,280
THOSE EYE DROKS ILLEGALS

1239
00:46:52,280 --> 00:46:54,080
RADICALS AND MDGIT IS A MIKE

1240
00:46:54,080 --> 00:46:54,800
STUDIES OF MULTIPLE ENDOCRINIA

1241
00:46:54,800 --> 00:46:57,560
GRIEWT O THE IPAD OWN AS A

1242
00:46:57,560 --> 00:46:58,880
TRANSFER ACE GREENE GENE SO BOTH

1243
00:46:58,880 --> 00:47:02,000
OF THESE ARE SIMILAR TO GPX 4 IN

1244
00:47:02,000 --> 00:47:03,720
THAT THEY'RE REMOVING THE OXYGEN

1245
00:47:03,720 --> 00:47:05,360
RADICALS FROM THE LOW SYSTEM.

1246
00:47:05,360 --> 00:47:07,160
ON THE LOWER LEFT ON THE SLIDE,

1247
00:47:07,160 --> 00:47:10,240
WHAT WE'VE DONE IS WE'VE

1248
00:47:10,240 --> 00:47:11,520
COMPARED THE MUTANT P53

1249
00:47:11,520 --> 00:47:14,400
EXPRESSING CELLS AND THOSE WITH

1250
00:47:14,400 --> 00:47:15,600
MUTANT P53 DEPLETION AND

1251
00:47:15,600 --> 00:47:19,200
BASICALLY VALIDATED THE DATA

1252
00:47:19,200 --> 00:47:21,320
FROM RNA SEQ, SO DEPLETION OF

1253
00:47:21,320 --> 00:47:24,040
YOU HADITANT P53 CAUSES DOWN

1254
00:47:24,040 --> 00:47:28,080
REGULATION OF THESE 2 IMPORTANT

1255
00:47:28,080 --> 00:47:31,120
GLUTEO THIGH OWN ENZYMES.

1256
00:47:31,120 --> 00:47:39,720
SO IN THESE CELLS, NOW, IN SIN

1257
00:47:39,720 --> 00:47:41,680
GENERATED EIC MICE, WE'VE

1258
00:47:41,680 --> 00:47:43,640
KNOCKED DOWN PRDX 6 AND THE

1259
00:47:43,640 --> 00:47:44,960
KNOCK DOWN DATAA ARE ON THE LEFT

1260
00:47:44,960 --> 00:47:46,800
AND WE GOT PRETTY GOOD KNOCK

1261
00:47:46,800 --> 00:47:48,720
DOWN ASK ON THE RIGHT WE MONITOR

1262
00:47:48,720 --> 00:47:49,040
TUMOR GROWTH.

1263
00:47:49,040 --> 00:47:50,760
SO CAN YOU CLEARLY SEE IN THE

1264
00:47:50,760 --> 00:47:52,760
RED AND GREEN BARS THAT IF WE

1265
00:47:52,760 --> 00:47:55,720
KNOCK DOWN THESE 2 ENZYMES WE'VE

1266
00:47:55,720 --> 00:47:59,560
SENSITIZED THE CELLS TO

1267
00:47:59,560 --> 00:47:59,880
FERROPITOSEIS.

1268
00:47:59,880 --> 00:48:10,440
SO WHAT WE THINK P53 IS DOING IS

1269
00:48:17,080 --> 00:48:18,200
BASICALLY BY ALLOWING EXPRESSION

1270
00:48:18,200 --> 00:48:19,960
OF THESE 2 GENES IT'S

1271
00:48:19,960 --> 00:48:21,000
MAINTAINING THE STAINING OF THE

1272
00:48:21,000 --> 00:48:24,000
PEROX DAITIONZ OF THE LIPID

1273
00:48:24,000 --> 00:48:26,840
RADICALS IS TOLERATED BY THE

1274
00:48:26,840 --> 00:48:27,040
CELL.

1275
00:48:27,040 --> 00:48:30,520
SO IF YOU REMEMBER THE GAIN OF

1276
00:48:30,520 --> 00:48:32,600
FUNCTION SLIDE, 1 OF THE WAYS

1277
00:48:32,600 --> 00:48:34,120
THAT MUTANT P53 CAN CONTRIBUTE

1278
00:48:34,120 --> 00:48:36,160
TO GAIN OF FUNCTION IS BASICALLY

1279
00:48:36,160 --> 00:48:37,880
BY INTERACTING WITH OTHER

1280
00:48:37,880 --> 00:48:40,560
TRANSCRIPTION FACTORS.

1281
00:48:40,560 --> 00:48:46,560
SO WE TOOK OUR RNA SEQ DATA AND

1282
00:48:46,560 --> 00:48:47,560
WE ASKED--THERE'S DIFFERENT

1283
00:48:47,560 --> 00:48:49,480
DATABASES THAT YOU CAN USE TO

1284
00:48:49,480 --> 00:48:51,560
BASICALLY ASK, IS THERE ANY

1285
00:48:51,560 --> 00:48:56,040
SIMILARITY BETWEEN ALL OF THESE

1286
00:48:56,040 --> 00:48:57,680
TRANSCRIPTOMES AND SO WHAT WE

1287
00:48:57,680 --> 00:49:03,040
IDENTIFIED IN THIS CASE IS 4

1288
00:49:03,040 --> 00:49:04,440
DIFFERENT--4 DIFFERENT PROMOTER

1289
00:49:04,440 --> 00:49:05,840
SEQUENCES THAT WERE PRESENT IN

1290
00:49:05,840 --> 00:49:09,920
THESE GENES THAT ARE UPREGULATED

1291
00:49:09,920 --> 00:49:10,760
UPON DEPLETION OF MUTANT P53,

1292
00:49:10,760 --> 00:49:12,160
AND THE 1 THAT CAUGHT OUR

1293
00:49:12,160 --> 00:49:16,360
ATTENTION IN THIS CASE, IS THIS

1294
00:49:16,360 --> 00:49:19,840
NRF 2 PROTEIN, AND NRF 2 IS A

1295
00:49:19,840 --> 00:49:21,200
DNA BINDING PROTEIN AND ABOUT A

1296
00:49:21,200 --> 00:49:24,360
THIRD OF THE GENES THAT WERE

1297
00:49:24,360 --> 00:49:26,480
DIFIENTIALLY EXPRESSED UPON

1298
00:49:26,480 --> 00:49:32,120
DEPLETION OF MUTANT P53 HAD AN

1299
00:49:32,120 --> 00:49:33,600
NRF 2 MOTIF.

1300
00:49:33,600 --> 00:49:36,080
AND NRF 2 IS IN THE

1301
00:49:36,080 --> 00:49:37,240
FERROPITOSEIS PATHWAY AND I WILL

1302
00:49:37,240 --> 00:49:38,600
SHOW YOU WHERE IT FITS IN A

1303
00:49:38,600 --> 00:49:39,000
MINUTE.

1304
00:49:39,000 --> 00:49:41,000
SO THE NEXT QUESTION WE ASK IS

1305
00:49:41,000 --> 00:49:42,720
BECAUSE THIS IS THE HYPOTHESIS

1306
00:49:42,720 --> 00:49:43,800
WE'RE WORKING WITH IS THAT

1307
00:49:43,800 --> 00:49:46,760
MUSEUM AT THAT POINT P53

1308
00:49:46,760 --> 00:49:48,080
INTERACTING WITH OTHER MUTANT

1309
00:49:48,080 --> 00:49:48,880
TRANSCRIPTION FACTORS TO DRIVE

1310
00:49:48,880 --> 00:49:50,440
THE PROGRAM SO THESE ARE 2

1311
00:49:50,440 --> 00:49:51,720
DIFFERENT ASSAYS IN WHICH WE

1312
00:49:51,720 --> 00:49:54,040
LOOK TO SEE IF MUTANT COULD

1313
00:49:54,040 --> 00:49:56,320
INTERACT WITH NRF 2, SO ON THE

1314
00:49:56,320 --> 00:49:59,120
LEFT IS IMMUNO PRECIPITATION

1315
00:49:59,120 --> 00:50:00,920
EXPERIMENTS IN CELLS, THAT

1316
00:50:00,920 --> 00:50:02,120
EXPRESS THE 172 MUTANT PROTEIN

1317
00:50:02,120 --> 00:50:04,920
AND YOU CAN SEE THAT IF WE

1318
00:50:04,920 --> 00:50:06,640
IMMUNO PRECIPITATE WITH P53 AND

1319
00:50:06,640 --> 00:50:09,440
THE RIGHT MOST LANE, WE CAN

1320
00:50:09,440 --> 00:50:10,840
DETECT NRF 2.

1321
00:50:10,840 --> 00:50:13,160
AND THEN ON THE RIGHT WAS SOME

1322
00:50:13,160 --> 00:50:16,520
INVITRO DATA WHERE BASICALLY WE

1323
00:50:16,520 --> 00:50:19,920
EXPRESS THESE PROTEINS, INVITRO

1324
00:50:19,920 --> 00:50:21,240
PURIFIED THEM AND AGAIN IN THE

1325
00:50:21,240 --> 00:50:22,520
RIGHT MOST PANEL YOU CAN SEE

1326
00:50:22,520 --> 00:50:25,080
THAT A EFTHIMIOS TAG NRF 2

1327
00:50:25,080 --> 00:50:27,960
PROTEIN IS INTERACTING WITH THE

1328
00:50:27,960 --> 00:50:29,760
245 W TAG.

1329
00:50:29,760 --> 00:50:31,600
SO, IN--AND WE'VE DONE THIS FOR

1330
00:50:31,600 --> 00:50:33,720
BOTH THE 245 AND THE 172 MUTANT

1331
00:50:33,720 --> 00:50:35,480
SO IT DOESN'T MATTER WHICH

1332
00:50:35,480 --> 00:50:37,640
MUTANT YOU'RE EXPRESSING IN THIS

1333
00:50:37,640 --> 00:50:41,120
SYSTEM, THAT MUTANT PROTEIN IS

1334
00:50:41,120 --> 00:50:42,840
INTERACTING WITH NRF 2.

1335
00:50:42,840 --> 00:50:45,960
SO, THIS IS A--THIS IS NOT A CHP

1336
00:50:45,960 --> 00:50:48,080
ASSAY, IT'S A NEW FANGLED CUT

1337
00:50:48,080 --> 00:50:49,560
AND RUN ASSAY WHERE AGAIN YOU'RE

1338
00:50:49,560 --> 00:50:53,560
ASKING THE SAME QUESTION, IS THE

1339
00:50:53,560 --> 00:50:55,440
MUTANT PROTEIN, MUTANT P53 AND

1340
00:50:55,440 --> 00:50:57,640
NR2 PRESCRIBINGENT ON THE SAME

1341
00:50:57,640 --> 00:50:59,920
DNA SEQUENCES SO WE TOOK 2 OF

1342
00:50:59,920 --> 00:51:04,200
THESE A. R. E. ELEMENTS FROM THE

1343
00:51:04,200 --> 00:51:06,600
PRDX 6 GENES, SITE 1 EXPW 2 AND

1344
00:51:06,600 --> 00:51:08,000
CAN YOU SEE THAT NRF 2 ON THE

1345
00:51:08,000 --> 00:51:09,440
LEFT IS BOUND TO THE SEQUENCES

1346
00:51:09,440 --> 00:51:12,080
AND ON THE RIGHT, YOU CAN SEE

1347
00:51:12,080 --> 00:51:14,080
THAT WHEN MUTANT P53 IS PRESENT,

1348
00:51:14,080 --> 00:51:16,360
IT'S ALSO BOUND TO THE SEQUENCE.

1349
00:51:16,360 --> 00:51:18,800
AND SO THIS WAS, I THINK VERY

1350
00:51:18,800 --> 00:51:19,840
CONVINCING DATA ALTHOUGH WE'RE

1351
00:51:19,840 --> 00:51:21,640
HAVING A HARD TIME CONVINCING

1352
00:51:21,640 --> 00:51:25,080
THE REVIEWERS THAT MUTANT P53 IS

1353
00:51:25,080 --> 00:51:25,800
INTERACTING WITH NRF 2.

1354
00:51:25,800 --> 00:51:27,880
SO HERE'S WHAT WE THINK IS

1355
00:51:27,880 --> 00:51:29,960
HAPPENING IN THE CELL, SO IN THE

1356
00:51:29,960 --> 00:51:33,320
LOWER LEFT HAND CORNER, YOU HAD

1357
00:51:33,320 --> 00:51:34,720
MUTANT P53 INTERACKING WITH NRF

1358
00:51:34,720 --> 00:51:37,160
2 AND IT'S DRIVING EXPRESSION OF

1359
00:51:37,160 --> 00:51:40,320
THESE 2 PROTEINS IN GST 3 AND

1360
00:51:40,320 --> 00:51:41,960
PRDX 6.

1361
00:51:41,960 --> 00:51:44,560
IN THESE PROTEINS, IN THE MIDDLE

1362
00:51:44,560 --> 00:51:46,680
GREEN, FUNCTION SIMILAR TO GPX

1363
00:51:46,680 --> 00:51:47,520
4, THEY'RE RESPONSIBLE FOR

1364
00:51:47,520 --> 00:51:50,680
REMOVING SOME OF THESE HYDROXY

1365
00:51:50,680 --> 00:51:52,080
RADICALS ON LIPIDS.

1366
00:51:52,080 --> 00:51:54,280
AND ALLOWING THE CELLS TO

1367
00:51:54,280 --> 00:51:54,560
SURVIVE.

1368
00:51:54,560 --> 00:51:55,920
SO WHAT HAPPENS IF YOU

1369
00:51:55,920 --> 00:51:56,840
TRANSITION TO 1 SIDE OF THE

1370
00:51:56,840 --> 00:51:58,800
SLIDE IS YOU REMOVE THE MUTANT

1371
00:51:58,800 --> 00:52:01,240
P53 AND YOU NO LONGER HAVE

1372
00:52:01,240 --> 00:52:02,480
EXPRESSION OF THESE 2 PROTEINS,

1373
00:52:02,480 --> 00:52:05,760
CAN YOU SEE FROM THE SINGLE CELL

1374
00:52:05,760 --> 00:52:06,560
DATA IN THE TRANSCRIPTOME DATA

1375
00:52:06,560 --> 00:52:11,800
THAT THESE 2 PROTEINS WERE DOWN

1376
00:52:11,800 --> 00:52:13,280
MODULATED UPON DOWN MODULATION

1377
00:52:13,280 --> 00:52:14,080
OF P53.

1378
00:52:14,080 --> 00:52:15,760
SO WE THINK YOU OFFSET THE

1379
00:52:15,760 --> 00:52:17,840
BALANCE, NOW YOU ONLY HAVE GPX 4

1380
00:52:17,840 --> 00:52:23,200
TRYING TO DEAL WITH THIS

1381
00:52:23,200 --> 00:52:24,440
INCREASED REACTIVE OXYGEN

1382
00:52:24,440 --> 00:52:25,400
SPECIES AND RADICALS AND IT'S

1383
00:52:25,400 --> 00:52:27,480
NOT SUFFICIENT AND SO REMOVING

1384
00:52:27,480 --> 00:52:34,680
THAT MUTANT P53 IS TAKING CELLS

1385
00:52:34,680 --> 00:52:35,080
DOWN THAT PATHWAY.

1386
00:52:35,080 --> 00:52:36,320
NOW 1 ARK DITIONAL SIDE STORY

1387
00:52:36,320 --> 00:52:38,320
THAT I WON'T GIVE YOU A WHOLE

1388
00:52:38,320 --> 00:52:40,160
LOT OF DETAILS BUT AGAIN IT'S

1389
00:52:40,160 --> 00:52:41,240
FASCINATING WITH RESPECT TO THE

1390
00:52:41,240 --> 00:52:42,960
COMPLEXITY OF THE SYSTEM, SO I

1391
00:52:42,960 --> 00:52:45,640
TOLD YOU AT THE BEGINNING THAT

1392
00:52:45,640 --> 00:52:47,600
THESE MICE GET THESE ADEN O

1393
00:52:47,600 --> 00:52:49,200
CARCINOMAS AND THEY ALSO GET

1394
00:52:49,200 --> 00:52:53,080
THESE SARCOMA LIKE TUMORS SO IN

1395
00:52:53,080 --> 00:52:55,520
BOTH INSTANCES WHEN WE REMOVE

1396
00:52:55,520 --> 00:52:59,360
THE MUTANT P53, WE GET DEPLETION

1397
00:52:59,360 --> 00:53:01,560
OF THE TUMOR VOLUME AND THE

1398
00:53:01,560 --> 00:53:04,240
ANIMALS LIVE LONGER BUT FOR THE

1399
00:53:04,240 --> 00:53:05,200
ADEN O CARCINOMAS, I'VE SHOWN

1400
00:53:05,200 --> 00:53:08,280
YOU ALL THE DATA FOR THE ADEN O

1401
00:53:08,280 --> 00:53:10,600
CARCINOMA, THIS SEEMS TO BE

1402
00:53:10,600 --> 00:53:12,360
FERROPITOSEIS, BUT IF WE LOOK AT

1403
00:53:12,360 --> 00:53:15,160
THE SARCOMA DATA IT'S NOT

1404
00:53:15,160 --> 00:53:16,240
FERROPITOSEIS, EVEN THOUGH THOSE

1405
00:53:16,240 --> 00:53:19,000
TUMORS THAT ARE DRIVEN BY THE

1406
00:53:19,000 --> 00:53:20,640
SAME P53 MUTATION, THEY DEVELOP

1407
00:53:20,640 --> 00:53:21,000
DIFFERENTLY.

1408
00:53:21,000 --> 00:53:23,400
THEY GIVE RISE TO A DIFFERENT

1409
00:53:23,400 --> 00:53:24,440
PATHOLOGY, AND WHILE THEY'RE

1410
00:53:24,440 --> 00:53:26,800
DEPEND OANT P53, THEY'RE NOT

1411
00:53:26,800 --> 00:53:28,560
DEPENDENT BECAUSE OF A

1412
00:53:28,560 --> 00:53:28,880
FERROPITOSEIS.

1413
00:53:28,880 --> 00:53:30,600
SO THAT JUST GIVES YOU 1 MORE

1414
00:53:30,600 --> 00:53:32,640
EXAMPLE OF THE COMPLEXITY OF

1415
00:53:32,640 --> 00:53:33,160
WORKING WITH P53.

1416
00:53:33,160 --> 00:53:39,040
I THINK THERE'S GOING TO BE A

1417
00:53:39,040 --> 00:53:41,720
LOT OF TISSUE SPECIFICITY.

1418
00:53:41,720 --> 00:53:43,480
SO HOPEFULLY I'VE GIVEN YOU A

1419
00:53:43,480 --> 00:53:46,840
SMATTERING OF SOME OF THE

1420
00:53:46,840 --> 00:53:48,120
STUDIES THAT WE'VE DONE.

1421
00:53:48,120 --> 00:53:50,720
I'M NASAA.COMINATED BY WILD-TYPE

1422
00:53:50,720 --> 00:53:51,080
P53 ACTIVITIES.

1423
00:53:51,080 --> 00:53:52,480
WHAT IS THE TRANSCRIPTIONAL

1424
00:53:52,480 --> 00:53:54,880
PROGRAM THAT P53 CAN INITIATE IT

1425
00:53:54,880 --> 00:53:56,040
IN DIFFERENT CELL TYPES.

1426
00:53:56,040 --> 00:53:57,600
WE ARE HOPING THAT MIGHT LEAD TO

1427
00:53:57,600 --> 00:54:00,080
THE ABILITY TO ACTIVATE THAT

1428
00:54:00,080 --> 00:54:01,840
PATHWAY DOWN STREAM AND

1429
00:54:01,840 --> 00:54:03,240
POTENTIALLY CONTRIBUTE TO TUMOR

1430
00:54:03,240 --> 00:54:04,480
SUPPRESSION IN SOME WAY?

1431
00:54:04,480 --> 00:54:07,120
AND THEN I TALKED ABOUT A LOT OF

1432
00:54:07,120 --> 00:54:08,760
THE DIFFERENT P53 MISSENSED

1433
00:54:08,760 --> 00:54:11,640
MUTATIONS, SOME OF THEM IN THE

1434
00:54:11,640 --> 00:54:12,320
GERM LINE, AND THEY'RE

1435
00:54:12,320 --> 00:54:13,520
DIFFERENT, THEY'RE NOT THE SAME

1436
00:54:13,520 --> 00:54:15,600
AND I THINK THE OTHER

1437
00:54:15,600 --> 00:54:17,440
FASCINATING ASPECT OF ALL OF

1438
00:54:17,440 --> 00:54:18,520
THESE MODELS THAT WE'VE

1439
00:54:18,520 --> 00:54:19,800
GENERATED IS WE NEVER KNOW WHAT

1440
00:54:19,800 --> 00:54:24,560
TUMOR TYPE WE'RE GOING TO GET.

1441
00:54:24,560 --> 00:54:26,320
SO I THINK TRYING TO UNDERSTAND

1442
00:54:26,320 --> 00:54:28,600
THE EVOLUTION OF THOSE TUMORS,

1443
00:54:28,600 --> 00:54:30,600
TRYING TO UNDERSTAND SOME OF THE

1444
00:54:30,600 --> 00:54:31,280
CONTRIBUTING LESIONS, AND SO

1445
00:54:31,280 --> 00:54:32,680
SOME OF THE STUDIES ONGOING IN

1446
00:54:32,680 --> 00:54:36,760
THE LAB ARE TRYING TO UNDERSTAND

1447
00:54:36,760 --> 00:54:38,240
THOSE EARLY LESIONS AND WHAT ARE

1448
00:54:38,240 --> 00:54:40,560
THE CHANGES THAT HAPPEN EARLY ON

1449
00:54:40,560 --> 00:54:41,640
THAT CONTRIBUTE TO DISEASE.

1450
00:54:41,640 --> 00:54:43,080
LIKE I SAID, WE'RE SEQUENCING A

1451
00:54:43,080 --> 00:54:46,040
BUNCH OF THESE TUMORS TO TRY TO

1452
00:54:46,040 --> 00:54:50,120
UNDERSTAND THE GENOME AS IT

1453
00:54:50,120 --> 00:54:54,160
EVOLVES, AND THEN LASTLY THERE

1454
00:54:54,160 --> 00:54:57,080
WAS 1 MORE, WE REALLY WANT TO

1455
00:54:57,080 --> 00:54:58,120
UNDERSTAND THE METASTATIC

1456
00:54:58,120 --> 00:54:59,200
PROCESS BECAUSE TAKEN--THEY

1457
00:54:59,200 --> 00:55:00,240
ULTIMATELY WHAT KILLS THE

1458
00:55:00,240 --> 00:55:04,360
PATIENT BESIDES DRUG RESISTANCE,

1459
00:55:04,360 --> 00:55:10,320
MICHAEL, AND SO, WE'RE

1460
00:55:10,320 --> 00:55:11,960
SEQUENCING THOSE IN THE LUNG AND

1461
00:55:11,960 --> 00:55:13,480
LIVER TO SEE WHAT CHANGES.

1462
00:55:13,480 --> 00:55:15,080
IT'S NOT AN EASY PROBLEM.

1463
00:55:15,080 --> 00:55:17,280
I THINK METASTASIS IS A VERY

1464
00:55:17,280 --> 00:55:18,920
DIFFICULT PROBLEM TO STUDY,

1465
00:55:18,920 --> 00:55:20,160
WE'VE STUDIED IT IN TISSUE

1466
00:55:20,160 --> 00:55:22,160
CULTURE WAY TOO LONG AND I THINK

1467
00:55:22,160 --> 00:55:25,400
WE NEED TO STUDY IT IN IN VIVO

1468
00:55:25,400 --> 00:55:26,680
SYSTEMS WHILE WATCHING THE

1469
00:55:26,680 --> 00:55:28,760
PROCESS BUT AGAIN WE NEED TO

1470
00:55:28,760 --> 00:55:30,440
MAKE THESE WITH TDTOMATO AND

1471
00:55:30,440 --> 00:55:31,640
SACRIFICE A LOT OF MICE EARLY IN

1472
00:55:31,640 --> 00:55:36,400
THE PROCESS TO TRY TO UNDERSTAND

1473
00:55:36,400 --> 00:55:37,240
THAT EVOLUTIONARY PROCESS, SO

1474
00:55:37,240 --> 00:55:39,440
WITH THAT, I WILL JUST KIND OF

1475
00:55:39,440 --> 00:55:41,640
THANK THE PEOPLE IN MY LAB, THIS

1476
00:55:41,640 --> 00:55:43,480
IS A NEW PICTURE EVER SINCE JUST

1477
00:55:43,480 --> 00:55:46,120
A FEW WEEKS AGO.

1478
00:55:46,120 --> 00:55:48,240
WE DROPPED THE MASK MANDATE AT

1479
00:55:48,240 --> 00:55:51,200
MD ANDERSON AND WE QUICK TOOK A

1480
00:55:51,200 --> 00:55:51,560
PICTURE.

1481
00:55:51,560 --> 00:55:53,720
[INDISCERNIBLE] DID ALL THE

1482
00:55:53,720 --> 00:55:55,840
STUDIES WITH THE FERROPITOSEIS

1483
00:55:55,840 --> 00:55:57,600
AND IT WAS REALLY HER WORK

1484
00:55:57,600 --> 00:55:58,920
LOOKING AT THE SEQUENCES,

1485
00:55:58,920 --> 00:56:01,440
LOOKING AT THE SAMPLES THAT

1486
00:56:01,440 --> 00:56:03,520
SUGGESTED THAT IT WAS A

1487
00:56:03,520 --> 00:56:05,720
MECHANISM BY WHICH MUTANT PPEA

1488
00:56:05,720 --> 00:56:11,200
WAS MAINTAINING THESE TUMORS,

1489
00:56:11,200 --> 00:56:13,840
[INDISCERNIBLE] IS A LONG-TERM

1490
00:56:13,840 --> 00:56:16,680
TECHNICIAN IN MY LAB, SYDNEY WHO

1491
00:56:16,680 --> 00:56:18,040
FINISHED PERPh.D. AND MOVED ON

1492
00:56:18,040 --> 00:56:20,320
SO WASN'T HERE FOR THE PICTURE

1493
00:56:20,320 --> 00:56:21,200
DID THE TRANSCRIPT OHM STUDIES

1494
00:56:21,200 --> 00:56:25,080
AND I WORK WITH 2 INCREDIBLE

1495
00:56:25,080 --> 00:56:26,120
PATHOLOGISTS, ADEL AND JAMES WHO

1496
00:56:26,120 --> 00:56:27,320
NEVER HESITATE TO LOOK AT TUMORS

1497
00:56:27,320 --> 00:56:28,200
THAT WE BRING THEM.

1498
00:56:28,200 --> 00:56:30,120
SO WITH THAT, I WILL STOP HERE

1499
00:56:30,120 --> 00:56:38,800
AND TAKE QUESTIONS.

1500
00:56:38,800 --> 00:56:39,600
[ APPLAUSE ]

1501
00:56:39,600 --> 00:56:41,360
>>THANK YOU, ANYONE'S
FREE TO

1502
00:56:41,360 --> 00:56:41,640
USE THE MIC.

1503
00:56:41,640 --> 00:56:43,200
LET ME START WITH A QUESTION

1504
00:56:43,200 --> 00:56:44,520
FROM ONLINE.

1505
00:56:44,520 --> 00:56:45,040
IT'S FROM [INDISCERNIBLE].

1506
00:56:45,040 --> 00:56:48,960
AND CAN YOU COMMENT ABOUT

1507
00:56:48,960 --> 00:56:51,640
ISOCHROMOSOMES 17 Q IN MYELOIDNY

1508
00:56:51,640 --> 00:56:52,520
O PLASMS.

1509
00:56:52,520 --> 00:56:54,440
THESE TUMORS ALWAYS LOSE 1 COPY

1510
00:56:54,440 --> 00:56:59,760
OF TP 53, DELETED, BUT WE NEVER

1511
00:56:59,760 --> 00:57:01,120
FOUND MUTATIONS IN THE SECOND

1512
00:57:01,120 --> 00:57:01,320
COPY.

1513
00:57:01,320 --> 00:57:03,120
THE BEHAVIORS OF THESE LEUKEMIA

1514
00:57:03,120 --> 00:57:08,800
ARE AS BAD AS THE TP53BIA

1515
00:57:08,800 --> 00:57:09,640
LEGALLIC LOST 1S?

1516
00:57:09,640 --> 00:57:11,520
>>YEAH, YEAH, SO I KNOW
THAT'S

1517
00:57:11,520 --> 00:57:13,200
TRUE, NONE OF OUR MODELS GET

1518
00:57:13,200 --> 00:57:15,160
LEUKEMIA, SO I THINK THAT

1519
00:57:15,160 --> 00:57:16,240
ESPECIALLY THE GERM LINE MODELS,

1520
00:57:16,240 --> 00:57:17,800
I THINK THAT THE OTHER CELL

1521
00:57:17,800 --> 00:57:20,080
TYPES ARE MORE SUSCEPTIBLE TO

1522
00:57:20,080 --> 00:57:21,120
P53 MUTATIONS IN THE BACKGROUNDS

1523
00:57:21,120 --> 00:57:23,480
THAT WE'RE WORKING WITH, IT'S A

1524
00:57:23,480 --> 00:57:24,520
FASCINATING PROBLEM AND I JUST

1525
00:57:24,520 --> 00:57:26,960
DON'T KNOW THE ANSWER.

1526
00:57:26,960 --> 00:57:27,520
IT'S DOING SOMETHING.

1527
00:57:27,520 --> 00:57:30,080
AND THOSE ARE THE WORST

1528
00:57:30,080 --> 00:57:31,600
LEUKEMIAS, WHEN THEY--WHEN THEY

1529
00:57:31,600 --> 00:57:35,040
HAVE A MUTANT P53, SO, AGAIN, I

1530
00:57:35,040 --> 00:57:36,440
THINK IT'S CONTEXT IS VERY

1531
00:57:36,440 --> 00:57:38,840
IMPORTANT AND I THINK WE NEED TO

1532
00:57:38,840 --> 00:57:40,360
LOOK AT THE TRANSCRIPT OHM IN

1533
00:57:40,360 --> 00:57:42,760
THE CONTEXT OF THAT MUTANT P53

1534
00:57:42,760 --> 00:57:43,640
AND HOW IT'S CHANGING.

1535
00:57:43,640 --> 00:57:46,560
BUT I'VE NOT KNOCKED ON AML BUT

1536
00:57:46,560 --> 00:57:49,760
I DO WANT TO.

1537
00:57:49,760 --> 00:57:51,920
WE'LL GET THERE.

1538
00:57:51,920 --> 00:57:53,920
>>YEAH, BEAUTIFUL.

1539
00:57:53,920 --> 00:57:56,920
AND LIKE AN IMMUNOLOGIST SO I

1540
00:57:56,920 --> 00:57:57,840
WOULD HAVE AN IMMUNOLOGY RELATED

1541
00:57:57,840 --> 00:57:58,240
QUESTION.

1542
00:57:58,240 --> 00:58:00,960
HAVE YOU LOOKED IN THIS TUMOR

1543
00:58:00,960 --> 00:58:02,720
INTO A RAG DEFICIENCY OR HOW

1544
00:58:02,720 --> 00:58:07,080
MUCH IS THE IMMUNE SYSTEM

1545
00:58:07,080 --> 00:58:07,440
IMPLICATED?

1546
00:58:07,440 --> 00:58:08,560
SO [INDISCERNIBLE] IT'S A VERY

1547
00:58:08,560 --> 00:58:10,600
LONG TIMELINE SO HAVE YOU ALSO

1548
00:58:10,600 --> 00:58:13,440
LOOKEDDA THE HOW MANY MUTATION

1549
00:58:13,440 --> 00:58:15,800
YOU ACCUMULATING IN THESE MODELS

1550
00:58:15,800 --> 00:58:18,160
AND HOW MANY CAN INDUCE A

1551
00:58:18,160 --> 00:58:19,360
SPECIFIC IMMUNE RESPONSE?

1552
00:58:19,360 --> 00:58:21,360
>>OKAY, SO WHAT WE'VE
DONE IS

1553
00:58:21,360 --> 00:58:22,920
WE'RE JUST IN THE PROCESS OF

1554
00:58:22,920 --> 00:58:24,200
SEQUENCING SO WE'RE GETTING ALL

1555
00:58:24,200 --> 00:58:25,880
THAT DATA BUT THE SINGLE CELL

1556
00:58:25,880 --> 00:58:31,280
DATA SUGGESTED THAT THERE WAS AN

1557
00:58:31,280 --> 00:58:33,480
IMMUNE RESPONSE WHEN WE REMOVE

1558
00:58:33,480 --> 00:58:34,760
THE MUTANT P53 SO WE'RE TRYING

1559
00:58:34,760 --> 00:58:36,160
TO FIGURE THAT OUT BUT WE

1560
00:58:36,160 --> 00:58:37,120
HAVEN'T DONE ANY EXPERIMENTS TO

1561
00:58:37,120 --> 00:58:39,880
ACTUALLY SAY HOW THAT IMMUNE

1562
00:58:39,880 --> 00:58:42,320
SYSTEM IS CONTRIBUTING TO THE

1563
00:58:42,320 --> 00:58:44,800
DECREASE IN THE TUMOR VOLUME AND

1564
00:58:44,800 --> 00:58:45,560
INCREASED SURVIVAL.

1565
00:58:45,560 --> 00:58:46,960
SO IN RELATION TO THAT, DO YOU

1566
00:58:46,960 --> 00:58:48,960
SEE LIKE IN HOW MANY CELLS, IS

1567
00:58:48,960 --> 00:58:52,200
THERE LIKE A TUMOR MOUSE WHICH

1568
00:58:52,200 --> 00:58:54,680
BECOMES CRITICAL IN TERMS OF

1569
00:58:54,680 --> 00:58:56,840
LIKE WHETHER THERE IS REJECTION

1570
00:58:56,840 --> 00:59:00,400
OR THERE IS LIKE REDUCTION OF

1571
00:59:00,400 --> 00:59:03,640
THE TUMOR GROWTH WHERE YOU

1572
00:59:03,640 --> 00:59:05,560
INDUCE, SO LIKE IF THE IMMUNE

1573
00:59:05,560 --> 00:59:06,840
RESPONSE IS INVOLVED, DO YOU

1574
00:59:06,840 --> 00:59:09,360
THINK THAT HOW MANY CELLS WOULD

1575
00:59:09,360 --> 00:59:11,480
THEN BE MUTATE SIDE RELEVANT?

1576
00:59:11,480 --> 00:59:12,600
>>YEAH, YEAH.

1577
00:59:12,600 --> 00:59:14,040
THAT'S A GREAT QUESTION AND 1 OF

1578
00:59:14,040 --> 00:59:16,040
THE REASONS THAT WE DEVELOPED

1579
00:59:16,040 --> 00:59:19,320
THE SOMATIC MODEL WITH THE ADEN

1580
00:59:19,320 --> 00:59:20,560
O DUCTAL INJECTION OF CREE IS

1581
00:59:20,560 --> 00:59:23,360
JUST TO MAKE A FEW CELLS BECOME

1582
00:59:23,360 --> 00:59:23,960
A TUMOR CELL.

1583
00:59:23,960 --> 00:59:25,600
SO I DON'T KNOW THE ANSWER TO

1584
00:59:25,600 --> 00:59:26,720
THE QUESTION, BUT I THINK WE CAN

1585
00:59:26,720 --> 00:59:29,000
USE THE MODEL TO TRY TO ADDRESS

1586
00:59:29,000 --> 00:59:29,720
THOSE QUESTIONS OVERTIME WHICH

1587
00:59:29,720 --> 00:59:33,840
IS I THINK WHAT YOU'RE ASKING,

1588
00:59:33,840 --> 00:59:34,040
YEAH.

1589
00:59:34,040 --> 00:59:34,280
>>GLEN?

1590
00:59:34,280 --> 00:59:35,360
>>GREAT TALK, SO I HAVE A

1591
00:59:35,360 --> 00:59:36,440
RELATED QUESTION OR MAYBE A

1592
00:59:36,440 --> 00:59:38,400
DEEPER QUESTION, SO YOU KNOW

1593
00:59:38,400 --> 00:59:40,400
YOU'VE DONE A FANTASTIC JOB OF

1594
00:59:40,400 --> 00:59:45,880
LOOKINGA THE INTRINSIC OR CELL

1595
00:59:45,880 --> 00:59:47,040
AUTONOMOUS MODELS BUT YOU COULD

1596
00:59:47,040 --> 00:59:48,000
ALMOST ELECTRIC AT EVERY PART

1597
00:59:48,000 --> 00:59:50,760
OFIOURE TALK AND ASK A QUESTION

1598
00:59:50,760 --> 00:59:51,840
ABOUT THE TUMOR

1599
00:59:51,840 --> 00:59:52,360
MICROENVIRONMENT.

1600
00:59:52,360 --> 00:59:54,960
SO FOR EXAMPLE, DO YOU KNOW IF

1601
00:59:54,960 --> 00:59:56,080
THE DIFFERENT MUTANTS HAVE

1602
00:59:56,080 --> 00:59:57,480
DIFFERENT EFFECTS ON EITHER THE

1603
00:59:57,480 --> 01:00:00,600
IMMUNE SYSTEM OR OTHER

1604
01:00:00,600 --> 01:00:01,200
MICROENVIRONMENTAL COMPONENTS?

1605
01:00:01,200 --> 01:00:02,120
DOES THAT PLAY A ROLE?

1606
01:00:02,120 --> 01:00:04,640
WHAT ABOUT AT THE METASTATIC

1607
01:00:04,640 --> 01:00:04,840
SITES?

1608
01:00:04,840 --> 01:00:06,320
AND OF COURSE WHEN YOU TURN IT

1609
01:00:06,320 --> 01:00:09,840
ON AND OFF, BUT THESE--THERE'S

1610
01:00:09,840 --> 01:00:10,640
PROBABLY DIFFERENCES.

1611
01:00:10,640 --> 01:00:11,280
BECAUSE THEY'LL CHANGE

1612
01:00:11,280 --> 01:00:13,080
SECRETIONS AND ALL KINDS OF

1613
01:00:13,080 --> 01:00:13,760
STUFF.

1614
01:00:13,760 --> 01:00:14,080
>>RIGHT.

1615
01:00:14,080 --> 01:00:15,520
WE'RE SEQUENCING ALL OF THESE

1616
01:00:15,520 --> 01:00:15,840
SAMPLES.

1617
01:00:15,840 --> 01:00:16,720
INVITE ME BACK NEXT YEAR.

1618
01:00:16,720 --> 01:00:22,080
NO, GIVE ME A COUPLE YEARS.

1619
01:00:22,080 --> 01:00:22,520
[LAUGHTER]

1620
01:00:22,520 --> 01:00:24,560
>>GIGI THANKS SO MUCH FOR
THE

1621
01:00:24,560 --> 01:00:25,080
LOVELY TALK.

1622
01:00:25,080 --> 01:00:28,160
SPEAKING ABOUT DRUG RESISTANCE,

1623
01:00:28,160 --> 01:00:32,120
1 PREDICTION IS THAT FOR YOUR

1624
01:00:32,120 --> 01:00:35,160
MUTANT P53, THAT HAS SOME WAY OF

1625
01:00:35,160 --> 01:00:35,720
BLOCKING FOR APOPTOSIS, YOU

1626
01:00:35,720 --> 01:00:37,280
WOULD EXPECT THAT THE TUMORS

1627
01:00:37,280 --> 01:00:42,000
WOULD BE MORE RESISTANT TO THE

1628
01:00:42,000 --> 01:00:44,480
NORMAL STIMULANTS, ARAFTIN FOR

1629
01:00:44,480 --> 01:00:45,880
EXAMPLE, FOR AFERROPITOSEIS, ARE

1630
01:00:45,880 --> 01:00:46,400
THEY RESISTANT TO THE

1631
01:00:46,400 --> 01:00:50,920
FERROPITOSEIS STIMULATORS?

1632
01:00:50,920 --> 01:00:54,640
>>SO WE HAVEN'T DONE THE
RAS

1633
01:00:54,640 --> 01:00:55,320
EXPERIMENT IN VIVO.

1634
01:00:55,320 --> 01:00:57,120
WE'VE ONLY DONE IT IN THE TISSUE

1635
01:00:57,120 --> 01:01:01,160
CULTURE CELLS, AT LEAST I DON'T

1636
01:01:01,160 --> 01:01:01,520
THINK SO.

1637
01:01:01,520 --> 01:01:04,080
>>IN TISSUE CULTURE ARE
THEY

1638
01:01:04,080 --> 01:01:05,120
RICYST ANT?

1639
01:01:05,120 --> 01:01:06,800
>>BUT THAT'S AN
EXPERIMENT WE

1640
01:01:06,800 --> 01:01:07,400
NEED DO.

1641
01:01:07,400 --> 01:01:09,040
>>AND WE HAD A
CONVERSATION

1642
01:01:09,040 --> 01:01:12,560
BEFORE ABOUT THE FACT IT THERE

1643
01:01:12,560 --> 01:01:16,160
ARE SOME MUTANT P53S THAT

1644
01:01:16,160 --> 01:01:16,960
IMSTIEWLT PGFG, MY FAVORITE

1645
01:01:16,960 --> 01:01:18,080
GENE, DO YOU KNOW IF THE 1

1646
01:01:18,080 --> 01:01:19,720
YOU'RE LOOKING AT IS A

1647
01:01:19,720 --> 01:01:21,120
STIMULATOR AND IF SO, THEN THE

1648
01:01:21,120 --> 01:01:24,280
CELLS SHOULD BE MORE GENERALLY

1649
01:01:24,280 --> 01:01:25,200
RESISTANT TO ANTICANCER?

1650
01:01:25,200 --> 01:01:25,600
>>RIGHT, RIGHT.

1651
01:01:25,600 --> 01:01:27,480
WE HAVE ALL THE SEQUENCING DATA

1652
01:01:27,480 --> 01:01:29,200
WE NEED TO LOOK AT FOR EXAMPLE

1653
01:01:29,200 --> 01:01:31,320
THOSE KINDS OF QUESTIONS.

1654
01:01:31,320 --> 01:01:32,200
YEAH.

1655
01:01:32,200 --> 01:01:33,360
YEAH.

1656
01:01:33,360 --> 01:01:33,800
YEAH.

1657
01:01:33,800 --> 01:01:35,480
>>HI, I CONQUER WITH
EVERYBODY

1658
01:01:35,480 --> 01:01:36,400
ELSE, WONDERFUL TALK, MY

1659
01:01:36,400 --> 01:01:38,040
QUESTION IS WHAT HAPPENS IN THE

1660
01:01:38,040 --> 01:01:41,600
CONTEXT OF A TUMOR VIRUS WHERE

1661
01:01:41,600 --> 01:01:44,440
YOU HAVE WILD-TYPE P53, AND THEN

1662
01:01:44,440 --> 01:01:46,880
A MUTANT THAT'S UNABLE TO ENGAGE

1663
01:01:46,880 --> 01:01:50,400
THE VIRAL PROTEIN LIKE T-ANTIGEN

1664
01:01:50,400 --> 01:01:52,200
OR WHAT HAVE YOU.

1665
01:01:52,200 --> 01:01:52,760
>>INTERESTING QUESTION.

1666
01:01:52,760 --> 01:01:55,680
SO YES, YES, SO TANTIGEN TUMOR

1667
01:01:55,680 --> 01:01:57,440
MODELS GENERATED YEARS AGO AND

1668
01:01:57,440 --> 01:01:58,680
IT WAS THE FIRST IN FACT THE

1669
01:01:58,680 --> 01:01:59,920
FIRST CANCER MODEL THAT WAS

1670
01:01:59,920 --> 01:02:05,920
GENERATED AND WE KNOW THAT

1671
01:02:05,920 --> 01:02:06,960
T-ANTIGEN SEQUESTERS WILD-TYPE

1672
01:02:06,960 --> 01:02:10,000
P53 PREVENTS IT FROM

1673
01:02:10,000 --> 01:02:11,400
FUNCTIONING, OKAY.

1674
01:02:11,400 --> 01:02:15,960
AND I DON'T REMEMBER EXACTLY

1675
01:02:15,960 --> 01:02:17,240
WHERE IN P53 T-ANTIGEN BINDS BUT

1676
01:02:17,240 --> 01:02:19,520
I'M GOING TO ASSUME IT'S THE

1677
01:02:19,520 --> 01:02:21,320
AMINO OR CAR BOXY TERMINUS

1678
01:02:21,320 --> 01:02:22,840
BECAUSE THAT'S WHERE MOST

1679
01:02:22,840 --> 01:02:24,840
INTERACTIONS OCCUR AND HAVE YOU

1680
01:02:24,840 --> 01:02:26,280
THE BYPASSING BOUND TO DNA AND

1681
01:02:26,280 --> 01:02:28,280
SO I THINK WHAT WOULD HAPPEN AND

1682
01:02:28,280 --> 01:02:30,080
I DON'T KNOW IF ANYBODY'S DONE

1683
01:02:30,080 --> 01:02:32,160
THE EXPERIMENT IS THAT THE

1684
01:02:32,160 --> 01:02:34,040
INTERACTIONS OF THAT T-ANTIGEN

1685
01:02:34,040 --> 01:02:36,120
WITH MUTANT P53 IS WOULD

1686
01:02:36,120 --> 01:02:37,160
STABILIZE THAT MUTANT PROTEIN

1687
01:02:37,160 --> 01:02:38,600
AND THEN YOU WOULD GET, IT

1688
01:02:38,600 --> 01:02:41,840
WOULDN'T JUST BE LOSS OF P53

1689
01:02:41,840 --> 01:02:43,360
WILD-TYPE P53 ACTIVITIES YOU

1690
01:02:43,360 --> 01:02:44,760
WOULD HAVE A MORE STABLE PROTEIN

1691
01:02:44,760 --> 01:02:46,400
THAT WOULD CONTRIBUTE TO A MORE

1692
01:02:46,400 --> 01:02:46,880
AGGRESSIVE PHENOTYPE.

1693
01:02:46,880 --> 01:02:48,000
IT WAS THE PREDICTION.

1694
01:02:48,000 --> 01:02:48,360
>>RIGHT.

1695
01:02:48,360 --> 01:02:52,280
AND THAT WOULD BE THE CASE FOR

1696
01:02:52,280 --> 01:02:54,440
EVERY ADEN O HPV WHARKS HAVE YOU

1697
01:02:54,440 --> 01:02:57,040
BECAUSE I NOTICED IN YOUR 125

1698
01:02:57,040 --> 01:03:02,080
GENES THAT WERE MODULATED,

1699
01:03:02,080 --> 01:03:04,800
CERVICAL, VIRAL AND BURKEETS.

1700
01:03:04,800 --> 01:03:05,240
>>RIGHT.

1701
01:03:05,240 --> 01:03:07,840
>>IN FACT WE PUBLISHED A
NICE

1702
01:03:07,840 --> 01:03:08,920
REVIEW A FEW YEARS BACK ABOUT

1703
01:03:08,920 --> 01:03:10,520
ALL THE MECHANISMS THAT CAN

1704
01:03:10,520 --> 01:03:11,600
ENACT VAIF P53 AND THAT'S

1705
01:03:11,600 --> 01:03:15,560
SOMETHING THAT WE NEED TO THINK

1706
01:03:15,560 --> 01:03:20,280
ABOUT BECAUSE YOU KNOW HPV, E6,

1707
01:03:20,280 --> 01:03:21,800
I THINK, DEGRADES WHILE TYPE P53

1708
01:03:21,800 --> 01:03:24,200
SO TO TREAT THOSE CANCERS YOU

1709
01:03:24,200 --> 01:03:26,000
NEED TO REMOVE HPV SO THAT THE

1710
01:03:26,000 --> 01:03:31,360
P53 CAN COME IN AND DO ITS WORK.

1711
01:03:31,360 --> 01:03:32,560
OTHER MECHANISMS INCLUDE

1712
01:03:32,560 --> 01:03:34,160
EXPRESSION OF THE MDM2 OR 4, AND

1713
01:03:34,160 --> 01:03:37,280
YOU SEE THOSE IN A LOT OF

1714
01:03:37,280 --> 01:03:38,080
SARCOMAS.

1715
01:03:38,080 --> 01:03:41,320
SO AGAIN, THERE ARE MDM2

1716
01:03:41,320 --> 01:03:42,040
INHIBITORS AND MDM4 BEING

1717
01:03:42,040 --> 01:03:47,760
DEVELOPED IN THE CLINIC TO

1718
01:03:47,760 --> 01:03:48,800
DISRUPT THAT MDM2 INTERACTION,

1719
01:03:48,800 --> 01:03:51,440
THEY'VE BEEN A LITTLE BIT TOXIC

1720
01:03:51,440 --> 01:03:53,120
IN THE TOXICITIES WERE PREDICTED

1721
01:03:53,120 --> 01:03:54,840
FROM OUR MODELS, BONE MARROW,

1722
01:03:54,840 --> 01:03:57,080
YOU KNOW IT'S A HIGHLY

1723
01:03:57,080 --> 01:03:59,600
PROLIFERATIVE TISSUE, YOU

1724
01:03:59,600 --> 01:04:00,440
DISRUPT THAT MDM2 P53

1725
01:04:00,440 --> 01:04:01,480
INTERACTION AND YOU DPET

1726
01:04:01,480 --> 01:04:05,360
DEPLETION OF BONE MARROW.

1727
01:04:05,360 --> 01:04:08,240
AND THEN, SO IT'S THE VIRAL

1728
01:04:08,240 --> 01:04:09,720
OVEREXPRESSED WITH P53 MUTATIONS

1729
01:04:09,720 --> 01:04:11,800
AND SEVERAL MECHANISMS AND I

1730
01:04:11,800 --> 01:04:14,600
THINK WE NEED TO BE COGNIZANT OF

1731
01:04:14,600 --> 01:04:17,520
HOW P53 WAS DISRUPTED IN ORDER

1732
01:04:17,520 --> 01:04:19,040
TO BE THINKING ABOUT HOW WE'RE

1733
01:04:19,040 --> 01:04:21,080
GOING TO TREAT THE PATIENTS BUT

1734
01:04:21,080 --> 01:04:22,960
WE'VE DONE A FEW STUDIES IN

1735
01:04:22,960 --> 01:04:31,880
ANIMAL MODELS WHERE WE

1736
01:04:31,880 --> 01:04:32,840
REINTRODUCED WILD-TYPE P53, IF

1737
01:04:32,840 --> 01:04:36,960
THERE'S NO P53 TO START WITH,

1738
01:04:36,960 --> 01:04:38,320
IT'S VERY GOOD AT SUPPRESSING

1739
01:04:38,320 --> 01:04:40,320
THAT GROWTH BUT IF YOU HAVE

1740
01:04:40,320 --> 01:04:41,720
MUTANT P53, IT CAN'T SUPPRESS.

1741
01:04:41,720 --> 01:04:43,720
, THATTA WHAT YOU HAVE WITH THE

1742
01:04:43,720 --> 01:04:43,960
VIRUS.

1743
01:04:43,960 --> 01:04:46,040
>>YES, SORE IF YOU HAVE
HIGH

1744
01:04:46,040 --> 01:04:47,680
LEVELS OF MDM2 YOU CAN'T GET

1745
01:04:47,680 --> 01:04:52,560
ENOUGH WILD-TYPE IN THERE TO

1746
01:04:52,560 --> 01:04:53,080
SUPPRESS GROWTH.

1747
01:04:53,080 --> 01:04:54,720
SO THOSE ARE ALL THINGS WE NEED

1748
01:04:54,720 --> 01:04:57,720
TO CONSIDER WHEN WE MOVE FORWARD

1749
01:04:57,720 --> 01:04:58,440
CLINICAL TREATMENTS.

1750
01:04:58,440 --> 01:04:59,600
>>YEAH.

1751
01:04:59,600 --> 01:05:01,240
>>THANKS.

1752
01:05:01,240 --> 01:05:01,520
>>YES?

1753
01:05:01,520 --> 01:05:03,800
>>I NOTICED THAT I MEAN
YOU

1754
01:05:03,800 --> 01:05:07,040
REALLY PRESENT THE STORY IN SOME

1755
01:05:07,040 --> 01:05:11,520
WAYS OF P53 AND PARTICULARLY THE

1756
01:05:11,520 --> 01:05:13,160
MUTANT AS A REDUCKS MODULATOR IN

1757
01:05:13,160 --> 01:05:15,480
THE CELL BUT I WONDER IF THE

1758
01:05:15,480 --> 01:05:18,120
OTHER SIDE OF THE EQUATION, THE

1759
01:05:18,120 --> 01:05:20,440
IRON AND THE HANDLING OF IRON

1760
01:05:20,440 --> 01:05:22,320
AND THE CELL, THE TRANSPORT OF

1761
01:05:22,320 --> 01:05:25,400
IRON INTO THE CELL OR OUT OF

1762
01:05:25,400 --> 01:05:27,760
TRANSFERRING DOES THAT CHANGE IN

1763
01:05:27,760 --> 01:05:29,920
ANY WAY WITH THESE MUTANTS.

1764
01:05:29,920 --> 01:05:31,600
>>THAT'S A GREAT
QUESTION.

1765
01:05:31,600 --> 01:05:34,560
WE HAVEN'T MEASURED IRON IN THE

1766
01:05:34,560 --> 01:05:34,800
SYSTEM.

1767
01:05:34,800 --> 01:05:36,240
I DON'T KNOW HOW TO DO IT BUT I

1768
01:05:36,240 --> 01:05:37,800
THINK WE COULD FIND OUT.

1769
01:05:37,800 --> 01:05:41,800
YEAH, THAT'S A GOOD QUESTION

1770
01:05:41,800 --> 01:05:47,960
BECAUSE, THAT IS REQUIRED, SO

1771
01:05:47,960 --> 01:05:50,880
MAYBE APPROACHING THE PROBLEM.

1772
01:05:50,880 --> 01:05:51,120
>>YEAH.

1773
01:05:51,120 --> 01:05:51,880
>>EQUATION THAT--

1774
01:05:51,880 --> 01:05:53,120
>>YEAH, I HADN'T THOUGHT
OF

1775
01:05:53,120 --> 01:05:58,480
THAT, THANK YOU.

1776
01:05:58,480 --> 01:05:58,840
>>THANK YOU.

1777
01:05:58,840 --> 01:06:00,360
>>WE WILL GO BACK AND
THINK

1778
01:06:00,360 --> 01:06:01,560
ABOUT EXPERIMENTS TO DO.

1779
01:06:01,560 --> 01:06:04,560
>>ONE MORE ONLINE, THIS
IS FROM

1780
01:06:04,560 --> 01:06:06,080
A COLLEAGUE ROWLAND OWEN WHO IS

1781
01:06:06,080 --> 01:06:10,160
I THINK TRIES TO MAKE ME TONGUE

1782
01:06:10,160 --> 01:06:11,200
TIED SO HERE'S MY BEST HERE

1783
01:06:11,200 --> 01:06:14,960
FLUTE O THE IPAD OWN CAN

1784
01:06:14,960 --> 01:06:16,120
RECHARGE NUCLEOTIDES O TIDE

1785
01:06:16,120 --> 01:06:18,880
REDUCT ACE, HAVE YOU LOOKEDDA

1786
01:06:18,880 --> 01:06:21,800
THE DNTPSPIEZ POOLS IN ANY OF

1787
01:06:21,800 --> 01:06:22,480
THESE P53 MUTANTS?

1788
01:06:22,480 --> 01:06:24,080
>>THAT'S A GREAT
QUESTION.

1789
01:06:24,080 --> 01:06:26,800
BECAUSE THERE'S OLD DAT THAT

1790
01:06:26,800 --> 01:06:28,640
WILD-TYPE P53 ACTIVATES REDUCT

1791
01:06:28,640 --> 01:06:29,720
ACE AND CAUSES CHANGES IN THE

1792
01:06:29,720 --> 01:06:31,440
DUCT ACE BUT WE DON'T HAVE

1793
01:06:31,440 --> 01:06:32,760
WILD-TYPE IN OUR SYSTEM, WE

1794
01:06:32,760 --> 01:06:37,800
HAVEN'T LOOKED SO WE NEED TO

1795
01:06:37,800 --> 01:06:38,000
LOOK.

1796
01:06:38,000 --> 01:06:38,160
YES?

1797
01:06:38,160 --> 01:06:40,960
>>I'M NOT SAYING THAT
WHAT

1798
01:06:40,960 --> 01:06:41,480
YOU'RE PRESENTING WASN'T

1799
01:06:41,480 --> 01:06:42,720
INTERESTING, I LOVED IT BUT I'M

1800
01:06:42,720 --> 01:06:48,040
STARTING TO GET CURIOUS ABOUT

1801
01:06:48,040 --> 01:06:52,680
THOSE NONP53 TUMORS AND IF

1802
01:06:52,680 --> 01:06:54,240
THERE'S A--IF YOU'VE SEEN OR A

1803
01:06:54,240 --> 01:06:54,960
POSSIBILITY THAT YOU INITTIAIT

1804
01:06:54,960 --> 01:06:57,360
THE TUMOR WITH THESE MUTANT

1805
01:06:57,360 --> 01:06:59,080
FORMS BUT THEN CAN YOU GET LOSS

1806
01:06:59,080 --> 01:07:07,480
OF THAT AND COMP PENSATORY LOSS

1807
01:07:07,480 --> 01:07:09,440
AND SAY FERRO--FER, OPITOSEIS.

1808
01:07:09,440 --> 01:07:12,160
DO YOU SEE IN THE TUMORS THERE'S

1809
01:07:12,160 --> 01:07:16,400
MUTATIONS IN IN FERROPITOSEIS

1810
01:07:16,400 --> 01:07:16,840
PATHWAY SNR.

1811
01:07:16,840 --> 01:07:17,600
>>THAT'S--WE DIDN'T
SEEQUENCE

1812
01:07:17,600 --> 01:07:19,200
THOSE TUMORS BUT I THINK THAT'S

1813
01:07:19,200 --> 01:07:21,400
A GOOD POINT BECAUSE YOU SHOULD

1814
01:07:21,400 --> 01:07:24,040
ALWAYS SEEQUENCE THE OPPOSITE,

1815
01:07:24,040 --> 01:07:25,160
THE 1S THAT AREN'T--NO, WE

1816
01:07:25,160 --> 01:07:26,720
HAVEN'T DONE THAT.

1817
01:07:26,720 --> 01:07:28,400
>>YOU JUST MIGHT SEE--

1818
01:07:28,400 --> 01:07:31,880
>>MAKES PERFECT SENSE,
YOU

1819
01:07:31,880 --> 01:07:32,960
DON'T JUST SEQUENCE THE 1S THAT

1820
01:07:32,960 --> 01:07:35,680
GIVE YOU THE RESULTS YOU WANT,

1821
01:07:35,680 --> 01:07:37,080
YOU SEQUENCE THEM TO GLEAN MORE

1822
01:07:37,080 --> 01:07:38,240
FROM THE 1S YOU DON'T.

1823
01:07:38,240 --> 01:07:39,440
WE WILL GO BACK.

1824
01:07:39,440 --> 01:07:42,280
, IT MAY COST MORE MONEY.

1825
01:07:42,280 --> 01:07:42,640
>>THANK YOU.

1826
01:07:42,640 --> 01:07:44,280
>>GIGI, THANK YOU
REFLECTIVE OF

1827
01:07:44,280 --> 01:07:46,480
MY IGNORANCE, IN THE FIELD OF

1828
01:07:46,480 --> 01:07:46,680
P53,.

1829
01:07:46,680 --> 01:07:48,560
>>ALL QUESTIONS ARE GOOD.

1830
01:07:48,560 --> 01:07:52,000
>>IS THERE ANY ROLE IN
STEM

1831
01:07:52,000 --> 01:07:53,840
CELL HOMEOSTASIS AND EACH STEM

1832
01:07:53,840 --> 01:07:57,040
CELL, P53 TREAT ANY ROLE?

1833
01:07:57,040 --> 01:07:59,040
AND I'M THINKING OF THOSE TUMORS

1834
01:07:59,040 --> 01:08:01,200
THAT ARE HYPOTHESIZED TO BE

1835
01:08:01,200 --> 01:08:05,280
DERIVED FROM STEM CELL AG

1836
01:08:05,280 --> 01:08:05,520
GRAVATION?

1837
01:08:05,520 --> 01:08:06,800
>>SO THE P53 NULL MOUSE
FOR THE

1838
01:08:06,800 --> 01:08:09,680
MOST PART IS VIABLE, SO AND IT

1839
01:08:09,680 --> 01:08:11,920
LOOKS NORMAL IN THE RIGHT SIZE

1840
01:08:11,920 --> 01:08:14,440
AND IT'S TUMOR PRONE, BUT IT

1841
01:08:14,440 --> 01:08:15,760
LOOKS DEVELOPMENT NORMAL SO IN

1842
01:08:15,760 --> 01:08:22,400
THAT SCENARIO, I WOULD SAY NO

1843
01:08:22,400 --> 01:08:24,800
BUT THERE'S AN INTERESTING

1844
01:08:24,800 --> 01:08:26,640
PHENOTYPE IN THE LIVER, AND THAT

1845
01:08:26,640 --> 01:08:28,880
IS IN YOU DELETE MDM2 IN THE

1846
01:08:28,880 --> 01:08:30,560
LIVER AND I TOLD YOU IT WAS CELL

1847
01:08:30,560 --> 01:08:37,040
LETHAL SO IT WAS--IT WAS TD THE

1848
01:08:37,040 --> 01:08:38,560
ALBUT MIN PROMOTER SO YOU DELETE

1849
01:08:38,560 --> 01:08:40,840
IT IN THOSE CELLS AND WHAT

1850
01:08:40,840 --> 01:08:42,680
HAPPENS IN THAT SCENARIO IS THAT

1851
01:08:42,680 --> 01:08:43,760
THE REMAINING CELLS REGENERATE A

1852
01:08:43,760 --> 01:08:43,960
LIVER.

1853
01:08:43,960 --> 01:08:46,160
SO THEY GET RID OF THE CELLS

1854
01:08:46,160 --> 01:08:48,960
THAT HAVE 2 HIGH LEVELS OF P53,

1855
01:08:48,960 --> 01:08:51,360
BUT THAT SENDS A SIGNAL TO THE

1856
01:08:51,360 --> 01:08:56,680
REST OF THE LIVER TO REGENERATE.

1857
01:08:56,680 --> 01:08:58,800
AND SO I DO THINK THERE'S A

1858
01:08:58,800 --> 01:08:59,840
WILD-TYPE ROLE IN REGENERATION.

1859
01:08:59,840 --> 01:09:01,920
I THINK THAT IT'S--IT'S NOT AT

1860
01:09:01,920 --> 01:09:03,600
THE CELL LEVEL LIKE YOU WERE

1861
01:09:03,600 --> 01:09:07,480
IMPLYING, IT'S AT THE ORGANISMAL

1862
01:09:07,480 --> 01:09:07,880
LEVEL.

1863
01:09:07,880 --> 01:09:09,920
WE HAD SIMILAR DATA AGING AGO IN

1864
01:09:09,920 --> 01:09:12,280
THE INTESTINE BECAUSE WE WOULD

1865
01:09:12,280 --> 01:09:13,760
TAKE OUT MDM2 IN THE INTESTINE

1866
01:09:13,760 --> 01:09:15,640
AND IN THE YOUNG MOUSE WE PUNCH

1867
01:09:15,640 --> 01:09:17,320
ALL THESE HOLES IN THE

1868
01:09:17,320 --> 01:09:19,040
INTESTINE, THEY WERE SICKLY, A

1869
01:09:19,040 --> 01:09:20,000
WEEK LATER THEY'RE FINE AND WHEN

1870
01:09:20,000 --> 01:09:22,800
WE LOOK AT THE MOUSE, EVERY CELL

1871
01:09:22,800 --> 01:09:25,920
THAT HAD DEPLETED MDM2 WAS GONE

1872
01:09:25,920 --> 01:09:27,480
AND WE HAD THIS CRYPT FUNCTION

1873
01:09:27,480 --> 01:09:30,640
THAT INCREASED AND SO,--SO THERE

1874
01:09:30,640 --> 01:09:34,640
IS A COMP PENSATORY MECHANISM TO

1875
01:09:34,640 --> 01:09:38,560
REGENERATE A TISSUE WHEN P53

1876
01:09:38,560 --> 01:09:39,480
DEPLETES THE TISSUE.

1877
01:09:39,480 --> 01:09:40,440
>>THANK YOU.

1878
01:09:40,440 --> 01:09:42,280
>>IT'S A FASCINATING
PHENOTYPE.

1879
01:09:42,280 --> 01:09:44,160
>>SO PLEASE JOIN ME IN

1880
01:09:44,160 --> 01:09:45,280
CONGRATULATING OUR SPEAKERS AND

1881
01:09:45,280 --> 01:09:47,920
THANKING THEM FOR A WONDERFUL

1882
01:09:47,920 --> 01:09:48,280
TALK.

1883
01:09:48,280 --> 01:09:49,120
[APPLAUSE ]

1884
01:09:49,120 --> 01:09:50,920
>>THANK YOU ALL.

1885
01:09:50,920 --> 01:10:01,440
ENJOYED BEING HERE IN PERSON.