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IT'S MY PLEASURE TO WELCOME

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DR. STEPHANIE HELENA TO THE

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NATIONAL CANCER INSTITUTE.

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DR. HELENA IS THE PROFESSOR OF

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HEMEATOLOGY AND CHIEF OF THE

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SECTION OF HEMEATOLOGY AT THE

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YALE SCHOOL OF MEDICINE AND I

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IPT GREATER VIETED HER BECAUSE I

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CONSIDER HER A STUNNING EXAMPLE

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OF A PHYSICIAN SCIENTIST WHOSE

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BOTH A CARING PHYSICIAN AND AN

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OUTSTANDING SCIENTIST WHO

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COMBINED BASIC TRANSLATIONAL AND

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CLINICAL STUDIES TO ADVANCE OUR

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UNDERSTANDING OF HEMEAT O LOGIC

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MALIGNANCIES.

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SHE RECEIVED HER DOCTORAL DEGREE

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FROM THE EVERHART MEDICINE AND

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AFTER THAT SHE JOINED CHILDREN'S

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HOSPITAL LOS ANGELES WHERE SHE

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STUDIED HEMATOPOIETIC STEM CELL

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BIOLOGY.

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IN 2000 SHE JOINED THE YALE

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INTERNAL MEDICINE RESIDENCY

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PROGRAM IN THE PHYSICIAN

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SCIENTIST RESEARCH TRACK AND IN

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2002 SHE BEGAN A HEMEATOLOGY

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FELLOWSHIP WHERE SHE STUDIED

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NUTRIFILL WITH MARY, AND THEN

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ALSO STUDIED WITH DR. KRAUSE.

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IN 2010 SHE TEABED HER OWN LAB

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RAARE TOY IN THE SECURITIZATION.

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OF HEMEATOLOGY AND IN 2019 SHE

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BECAME THE INTERIM SECTION CHIEF

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OF HEMEATOLOGY AND IN 2020, WAS

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LABEL--WAS NAMED SECTION CHIEF,

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WAS NAMED THE SECTION CHIEF OF

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HEMEATOLOGY.

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SHE ESTABLISHED THE HEMEATOLOGY

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TISSUE BANK AND TOOK ON THE ROLE

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OF ASSISTANT MEDICAL DIRECTOR OF

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THE CLINICAL TRIALS RESEARCH

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SUPPORT LABORATORY IN THE YALE

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CANCER CENTER.

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SHE'S ALSO CHIEF OF THE DIVISION

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OF CHANSALATION HEMEATOLOGY AND

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DIRECTS THE DE LUCA CENTER FOR

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HEMEATOLOGY RESEARCH IN THE YALE

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CANCER CENTER.

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HER SCIENTIFIC INTERESTS FOCUS

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ON HEMEAT O POETICESIS,

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MYELODYSPLASTIC SYNDROME AND

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AAMERICA L.

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AND IN COLLABORATION WITH

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COLLEAGUES AT YALE AND ELSEWHERE

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SHE'S DEVELOPED A

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XENOTRANSPLANTATION MODEL FOR

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MDS, ADVANCED OUR UNDERSTANDING

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OF R NA SPLICING FACTORS IN MDS

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AND ALSO HEMEAT ON POETICESIS

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AND LEUKEMIA.

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SHE'S RECEIVED SIGNIFICANT

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RECOGNITION FOR HER WORK.

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IN 2015 SHE RECEIVED THE SIR

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WILLIAM AWARD FROM THE

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INTERURBAN CLINICAL CLUB AND IN

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2022 SHE RECEIVED THE EMERGING

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LEADER IN INTERNAL MEDICINE

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AWARD FROM THE YALE SCHOOL OF

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MEDICINE AND IN 2024 SHE WAS

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ELECTED TO MEMBERSHIP IN A

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CONNECTICUT ACADEMY OF SCIENCE

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AND ENGINEERING SO PLEASE JOIN

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ME IN WELCOMING DR. STEPHANIE

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HELENA TO THE NCI.

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[ APPLAUSE ]

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>> THANK YOU SANDY FOR
THIS

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AMAZING INTRODUCTION, VERY KIND.

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[INDISCERNIBLE] COVID OR FLU,

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BUT OTHERWISE IT'S JUST

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ALLERGIES AND THEY DON'T SEEM TO

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GO AWAY, BUT I UPON TO SAY AS A

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PHYSICIAN, I REALLY, REALLY YOU

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KNOW WHEN I STARTED RESEARCH, I

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REALLY HAD NO CLUE AND I THINK

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TO BE A PHYSICIAN WHO ALWAYS

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DOES RESEARCH IT TAKES THIS

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VILLAGE AROUND YOU AND SANDY

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[INDISCERNIBLE] WAS 1 OF THOSE

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PEOPLE ENCOURAGING PHYSICIANS

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LIKE ME, YEAH, JUST WORK ON RNA,

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IT'S AN ARK MAZING FIELD AND

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THAT IS I'M INCREDIBLY GRATEFUL

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FOR THE SCIENTISTS, INVITING

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SOMEBODY LIKE ME INTO DOING

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SCIENCE AND DISEASE, SO I HAVE

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KIND OF 2 STORIES FOR YOU AND I

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KNOW THIS IS ALSO CLINICAL, I

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HAVE A BIT OF CLINICAL

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BACKGROUND.

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I KNOW THERE ARE TRAINEES IN THE

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ROOM, IF YOU WANT TO ASK

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QUESTIONS, PLEASE INTERRUPT ME

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AND WE CAN IMPROVISE AT THE END

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AND CUT A FEW SLIDES.

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OKAY, LET'S SEE HOW I MOVE

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FORWARD.

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>> OKAY, SO MY LAB IN
PARTICULAR

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IS INTERESTED IN MYELODYSPLASIA

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BECAUSE IT'S ALSO A CLINICAL

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PROBLEM I'M INTERESTED WHEN I

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SEE THESE PATIENTS I ALSO WANT

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TO KNOW WHAT WE CAN DO FOR THEM

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AND TREATMENT ISN'T GREAT AND IF

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YOU LOOK HERE AT MDS

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MYELODYSPLASIA, THERE'S OVER

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20,000 PATIENTS IN THE U.S. PER

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YEAR, IT'S CAUSED BY GENETIC,

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EPIGENETIC, TRANSKRINGZ AND

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SIGNALING AND SPLICING

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ABNORMALITIES AND MANY PATIENTS

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HAVE CYTOPENIA, AND HEART

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DISEASE OR WHATEVER DUE TO MDS

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AND A SUBSET PROGRESS TO AML.

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IF YOU SEE HERE, SURVIVAL ISN'T

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GREAT AND IT AFFECTS OLDER

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PEOPLE ON THE RIGHT THAT IS

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ALWAYS RELATIVE.

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SO THE HEMATOPOIETIC STEM CELL

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DISORDER SO ALL CELLS IN THE

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BLOOD CAN BE AFFECTED BUT

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PREDOMINANTLY MYELOID

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ILLUMINATEIAGE IS SHOWN HERE

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ASSOCIATED WITH INCREASED AGE,

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EXPOSURE TO PREVIOUS CHEMO

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THERAPY AND RADIATION, SO IF YOU

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CURE PATIENTS FROM THE CANCER,

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THEY'RE NOW AT RISK FOR MDS, AND

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THEN PROGRESSION TO SECONDARY

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AML AND IN 2011, BEING AN

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OPPORTUNE TIME FOR MY LAB, THESE

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MUTATIONS WERE DISCOVERED TO

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OCCUR IN OVER 50% OF THE

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PATIENTS WITH MDS AND SEVERAL

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OTHER PATHWAYS ARE ALSO AFFECTED

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BY OUR FOLKS TODAY ARE ON

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SPLICING.

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SO MY TALK HAS KIND OF 2 SIDES,

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1 IS REALLY TALKING ABOUT OUR

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WORK ON LIMITATIONS AND I HOPE I

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WILL GET TO TELL YOU ABOUT THE

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XENOTRANSPLANTATION MODEL WE

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DEVELOPED TOGETHER WITH RICHARD

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[INDISCERNIBLE].

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SO JUST A FACTOR ON THE

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MUTATIONS, THEY WERE DISCOVERED

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IN 2011, YOU WONDERED WHY NOT

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BEFORE BUT THIS WAS A BIG EFFORT

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TO SEQUENCE THE MDS GENOME, THEY

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ARE REALLY KEY FACTORS OF THE

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SPLICING MACHINERY AND MAYBE

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THAT'S WHY THEY WERE ALSO KIND

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OF--LIMITATIONS WERE IGNORED

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BECAUSE PEOPLE THOUGHT IT WAS

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IMPOSSIBLE.

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AND THEY OCCURRED IN OVER 50% OF

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THE PATIENTS WITH MDS OR CHRONIC

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MONOSITTIC LEUKEMIA AND IN 20%

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OF THE PATIENTS WITH SECONDARY

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AML BUT ALSO WITH THE CHRONIC

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LEUKEMIA AND EVEN THIS A HANDFUL

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OF SOLID TUMORS AND WE DON'T

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KNOW WHY THE SELECTION SO

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THERE'S STILL A LOT OF WORK ON

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TO DO.

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WHAT'S INTERESTING IS THAT I

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THEY ARE MUTUALLY EXCLUSIVE SO

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PATIENTS GENERALLY OVER HAVE 1

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OF THE PLIESING MUTATIONS, IF

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THERE ARE 2, THEY ARE IN THE

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SAME CELL, THE VERY FREQUENTY IS

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HIGH, AND REALLY THEY ARE IN THE

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DOMINANT CLONE, AND WE KNOW THAT

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THEIR DRIVER MUTATIONS FOR MANY

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MOUSE STUDIES FOR EXAMPLE, SO

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JUST A BRIEF, BRIEF, VERY SIMPLE

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PRIMER ON SPLICING RIGHT, SO IN

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THE GENE, YOU HAVE REGULATORY

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SEQUENCES, ENHANCES PROMOTERS

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YOU HAVE THE TEAM AND THEN THE

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YEEN IS EXIT AND ENTRON, THEY

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SPLICE THESE ENTRONS OUT TO

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CREATE THE MRNA, OR THE

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RNATHAT'S UNCAPPED,

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[INDISCERNIBLE] EXPORTED INTO

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THE CYTOPLASM WHERE IT'S THEN

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TRANSLATED INTO PROTEINS.

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AND YOU KNOW AUTOSPLICING IS AN

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AMAZING REGULATORY MECHANISM

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WHERE YOU CAN NOW CREATE PROTEIN

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DIVERSITY FROM A TRANSCRIPT

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THAT'S DIFFERENTIALLY SPLICED

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WHEN THE SPLICING IS

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DISREGULATIONED, THEN YOU KNOW

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YOU LOSE PROTEINS, YOU GAIN

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PROTEINS, SO THAT IS PART OF THE

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DISEASE PROCESS.

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GENERALLY WE THINK OF THESE

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MUTATIONS AS HOT SPOT MUTATIONS

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SO THEY'RE VERY SPECIFIC

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RESIDUES IN THE DIFFERENT

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PROTEINS THAT MUTATED AND WHAT

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WE KNOW SO FAR IS THAT THEY ARE

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ALL KIND OF INTERACT WITH THE

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RNA, ZRSR2 IS A BIT DIFFERENT,

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THE CELLS HAVE TO HAVE A

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WILD-TYPE ALLELE AND WE THINK OF

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THEM AS GAIN OF MUTATION

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FUNCTIONS.

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MEANING THE PROTEIN FUNCTION IS

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LOT BUT THERE'S SOMETHING NEW

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ADDED TO WHAT THE PRACTICES TEEN

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- IS NOW DOING.

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AND WHEN WE SPLICE THESE

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MUTATIONS TO FIND CLINICAL MINO

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TYPES SO AS THE MUTATIONS OCCUR,

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THE CANNED WHAT UV MELANOMA AND

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MUTATIONS OCCUR IN MDS BUT IN

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LUNG CANCER AS THEY OCCUR IN MDS

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AND CHRONIC MILE O SIDIC

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LEUKEMIA BUT I DON'T THINK

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THIEVE BEEN SEEN INSIDE TUMORS.

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THIS IS JUST AN OLDER

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CLASSIFICATION, AND YOU CAN SEE

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HERE, YOU KNOW HOW FREQUENT

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THESE SPLICING FACTORS ARE, THE

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MUTATIONS ARE THE MOST COMMON

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AND THEY'LL CURE AGAIN, THERE'S

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SUBTYPE SPECIFICITY WITH AN MDS,

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I KIND OF TRIED TO PULL OUT LIKE

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THE KEY DIFFERENTIALLY PLIESED,

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YOU KNOW TRANSCRIPTS AND YOU

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KNOW RIGHT, THEY VARY BETWEEN

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THE DIFFERENT SPLICING FACTORS,

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THERE ARE FEW THAT MAYBE

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OVERLAPPING BUT OF COURSE WE ALL

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WANT TO KNOW WHY LIMITATIONS

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WHAT DO THEY HAVE IN COMMON AND

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MAYBE THAT'S A COMMON MECHANISM

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THAT WE CAN TARGET, AND WHAT YOU

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SEE HERE IT'S AGAIN, THIS TILL A

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SIMPLIFICATION OF PLIESING AND

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YOU VALID TO HAVE

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00:09:21,445 --> 00:09:24,081
[INDISCERNIBLE] IN THE AUDIENCE

255
00:09:24,081 --> 00:09:28,218
TO GO IN NOR DETAIL BUT YOU SEE

256
00:09:28,218 --> 00:09:29,286
THESE SPLICING FACTORS, ALL HAVE

257
00:09:29,286 --> 00:09:31,255
A FUNCTION EARLY IN THE WHOLE

258
00:09:31,255 --> 00:09:33,023
SPLICING PROCESS AS I DEFINE THE

259
00:09:33,023 --> 00:09:34,992
EXOHMS THAT ARE INCLUDED OR

260
00:09:34,992 --> 00:09:37,995
EXCLUDED, THE SPLICE SIDES, AND

261
00:09:37,995 --> 00:09:42,766
REALLY INITIAL ATTACK AT THESE

262
00:09:42,766 --> 00:09:44,401
ENTRON EXON JUNCTIONS.

263
00:09:44,401 --> 00:09:46,236
AND SO, EARLY ON, SO WE WERE

264
00:09:46,236 --> 00:09:49,073
WORKING ON REALLY, REALLY

265
00:09:49,073 --> 00:09:50,474
COMPLICATED FUSION PROTEIN IN MY

266
00:09:50,474 --> 00:09:52,676
LAB AND I WAS INTERESTING IN THE

267
00:09:52,676 --> 00:09:54,011
BINDING PROCESS AND THESE

268
00:09:54,011 --> 00:09:55,612
MUTATIONS WERE PUBLISHED AND

269
00:09:55,612 --> 00:09:58,148
THAT'S WHAT WE JUMPED ON BECAUSE

270
00:09:58,148 --> 00:10:01,585
I REALLY WANTED TO BE AN ADULT

271
00:10:01,585 --> 00:10:03,921
HEMEATOLOGYIST AND NOT A

272
00:10:03,921 --> 00:10:06,056
PEDIATRIC HEMEATOLOGYIST, AND WE

273
00:10:06,056 --> 00:10:07,091
CHOSE THE [INDISCERNIBLE]

274
00:10:07,091 --> 00:10:09,126
BECAUSE IT BINDS RNA, IT BINDS

275
00:10:09,126 --> 00:10:10,728
EXONS AND THOSE ARE MORE

276
00:10:10,728 --> 00:10:13,063
CONSERVED BETWEEN MOUSE AND

277
00:10:13,063 --> 00:10:13,997
MUSEUM MAN.

278
00:10:13,997 --> 00:10:16,600
AND JUST A QUICK PRIMER ON WHAT

279
00:10:16,600 --> 00:10:18,202
WE DID, RIGHT?

280
00:10:18,202 --> 00:10:20,204
, SO, AS THE TUMORS ARE KNOWN TO

281
00:10:20,204 --> 00:10:21,939
HAVE THE RNA BINDING DOMAIN AND

282
00:10:21,939 --> 00:10:23,841
KIND OF LIKE THESE PIECES OF RNA

283
00:10:23,841 --> 00:10:26,143
AND 1 SIDE AND ON THE OTHER SIDE

284
00:10:26,143 --> 00:10:27,544
AND THIS REGION HERE IS CALLED

285
00:10:27,544 --> 00:10:29,012
THE LINKER SO IT LOOKED LIKE

286
00:10:29,012 --> 00:10:30,881
THESE MUTATIONS ARE JUST IN A

287
00:10:30,881 --> 00:10:33,050
LINKER REGION, BUT THAT'S NOT

288
00:10:33,050 --> 00:10:37,254
TRUE BECAUSE YOU CAN SIGH HERE,

289
00:10:37,254 --> 00:10:39,123
THESE 2 RNA SEQUENCES, AND

290
00:10:39,123 --> 00:10:41,959
ACTUALLY IT'S A CONIF I WERATION

291
00:10:41,959 --> 00:10:45,696
CHANGES COME SF 2 WHEN RNA'S

292
00:10:45,696 --> 00:10:48,232
THERE AND THESE 2 LINKERS AND

293
00:10:48,232 --> 00:10:49,600
END TERMINAL REGION ARE REALLY

294
00:10:49,600 --> 00:10:51,301
KIND OF LIKE ARMS THAT GO AROUND

295
00:10:51,301 --> 00:10:52,469
THE RNA AND IT MAKES SENSE IF

296
00:10:52,469 --> 00:10:54,338
YOU HAVE A MUTATION HERE, A

297
00:10:54,338 --> 00:10:56,240
CHANGE MAY BE HOW RNA IS BOUND

298
00:10:56,240 --> 00:10:59,510
AND THAT'S WHEN WE DID IN

299
00:10:59,510 --> 00:11:00,778
COLLABORATION WITH YOUR

300
00:11:00,778 --> 00:11:02,246
[INDISCERNIBLE], AND GOT ON TO

301
00:11:02,246 --> 00:11:04,882
SHOW THAT THERE IS SIGNIFY

302
00:11:04,882 --> 00:11:06,717
SEQUENCE SPECIFIC DIFFERENCE IN

303
00:11:06,717 --> 00:11:08,218
BINDING AFFINITY.

304
00:11:08,218 --> 00:11:13,090
WE   PERFORMED YOU KNOW BACK

305
00:11:13,090 --> 00:11:15,526
THEN HITS CLIP AND NOT SO

306
00:11:15,526 --> 00:11:16,693
EFFICIENT TECHNOLOGY TO LOOK AT

307
00:11:16,693 --> 00:11:18,896
HOW WILD-TYPE AND MUTANT, HAVE

308
00:11:18,896 --> 00:11:21,331
BIND RNA AND THEN WE SEE A

309
00:11:21,331 --> 00:11:23,700
SEQUENCE MOTIVE DIFFERENCE AND

310
00:11:23,700 --> 00:11:27,471
SO IN MUTANT TO CC&  G

311
00:11:27,471 --> 00:11:29,940
CONTAINING EXONS ARE

312
00:11:29,940 --> 00:11:32,042
PREFERENTIALLY INCLUED AND

313
00:11:32,042 --> 00:11:32,910
[INDISCERNIBLE] ARE EXCLUDED.

314
00:11:32,910 --> 00:11:34,445
SO YOU HAVE ABERRANT SPLICING.

315
00:11:34,445 --> 00:11:38,849
AND THEN AT THE SAME TIME, WHILE

316
00:11:38,849 --> 00:11:41,051
BRADLEY AND DAUBNER WERE WORKING

317
00:11:41,051 --> 00:11:42,886
ON [INDISCERNIBLE], AND OMAR

318
00:11:42,886 --> 00:11:44,421
GENERATED THE MOUSE MODEL SO WE

319
00:11:44,421 --> 00:11:52,696
PUT ALL OUR DATA TOGETHER AND

320
00:11:52,696 --> 00:11:54,832
THE SRSF2 MUTATIONS DID BIND AND

321
00:11:54,832 --> 00:11:57,267
SPLICE, IT IS INCLUSION OF

322
00:11:57,267 --> 00:11:58,101
SOPHISTICATED CALLED POISON EXON

323
00:11:58,101 --> 00:11:59,636
THAT CHANGES THE FRAME AND

324
00:11:59,636 --> 00:12:04,942
RESULTS IN DECREASED TRANSLATION

325
00:12:04,942 --> 00:12:06,443
OF EZH2, ALSO CONSIDER IN

326
00:12:06,443 --> 00:12:09,413
MDECKER S BUT THEY'RE MUTT YULE

327
00:12:09,413 --> 00:12:11,415
LE EXCLUSIVE SO OTHER BIOLOGY

328
00:12:11,415 --> 00:12:13,050
MADE SENSE, I ALWAYS FELT THIS

329
00:12:13,050 --> 00:12:14,418
WAS LIKE THE FINAL PUBLISHING WE

330
00:12:14,418 --> 00:12:15,552
HAVE TO DO, RIGHT?

331
00:12:15,552 --> 00:12:17,754
YOU HAVE A REALLY COMPLEX

332
00:12:17,754 --> 00:12:19,857
PROCESS, YOU AND YOU FIND 1

333
00:12:19,857 --> 00:12:20,824
ALTERNATIVE SPLICED EVENT THAT

334
00:12:20,824 --> 00:12:22,659
TELLS A BEAUTIFUL STORY, SO WE

335
00:12:22,659 --> 00:12:25,362
STILL DON'T KNOW SO MUCH ABOUT

336
00:12:25,362 --> 00:12:26,597
HOW THE SPLICING FACTOR

337
00:12:26,597 --> 00:12:30,133
MUTATIONS FUNCTION BUT IT WAS A

338
00:12:30,133 --> 00:12:30,667
GREAT START.

339
00:12:30,667 --> 00:12:32,336
SO OF COURSE EARLY ON, YOU KNOW

340
00:12:32,336 --> 00:12:34,304
PEOPLE WERE ASKING, OKAY, CAN WE

341
00:12:34,304 --> 00:12:37,374
TREAT PATIENTS, CAN WE TREAT

342
00:12:37,374 --> 00:12:39,409
PATIENTS AND FROM THE FACT THAT

343
00:12:39,409 --> 00:12:42,746
YOU KNOW THAT THESE MUTATIONS

344
00:12:42,746 --> 00:12:44,081
ALWAYS HETEROZYGOUS, HE KNOWS HE

345
00:12:44,081 --> 00:12:47,551
RELIES ON THE WILD-TYPE ALLELE,

346
00:12:47,551 --> 00:12:49,186
THEY'RE MUTUALLY EXCLUSIVE SO

347
00:12:49,186 --> 00:12:51,021
ONLY 1 SPLICING MUTATION

348
00:12:51,021 --> 00:12:52,823
TOLERATED, LATER ON PEOPLE PROVE

349
00:12:52,823 --> 00:12:53,991
THAT IN-MOUSE MODELS AND SO CAN

350
00:12:53,991 --> 00:12:57,761
YOU ACTUALLY KIND OF LIKE, YOU

351
00:12:57,761 --> 00:12:59,396
KNOW PERTURB THE WILD-TYPE

352
00:12:59,396 --> 00:13:01,532
NORMAL SPLICING PROCESS AND WHAT

353
00:13:01,532 --> 00:13:04,134
MUTANT CELLS WOULD BE

354
00:13:04,134 --> 00:13:05,602
PREFERENTIALLY SENSITIVE.

355
00:13:05,602 --> 00:13:07,905
AND YOU KNOW THERE WAS AN

356
00:13:07,905 --> 00:13:09,540
INHIBITOR OF SF 3 B 1 AND SO

357
00:13:09,540 --> 00:13:11,241
PEOPLE THOUGHT LIKE OKAY, COME

358
00:13:11,241 --> 00:13:13,577
AND USE THE SFB 3 B 1 INHIBITOR

359
00:13:13,577 --> 00:13:17,247
AND THEN CAN WE SPECIFICALLY

360
00:13:17,247 --> 00:13:19,016
TARGET MUTANT MDS AND I THINK IN

361
00:13:19,016 --> 00:13:21,151
MOUSE MODELS AND CELL LINES IT

362
00:13:21,151 --> 00:13:21,952
LOOKED ALL GREAT.

363
00:13:21,952 --> 00:13:23,954
WHEN I WENT INTO CLINICAL TRIAL,

364
00:13:23,954 --> 00:13:27,291
UNFORTUNATELY IT DID NOT WORK.

365
00:13:27,291 --> 00:13:30,827
THERE'S ALSOA A LOT OF FEAR

366
00:13:30,827 --> 00:13:33,664
OVER, YOU KNOW TOXICITY, THIS

367
00:13:33,664 --> 00:13:35,499
WAS LIKE AN OLDER COMPOUND THAT

368
00:13:35,499 --> 00:13:36,867
HAD LED TO BLINDNESS WHICH

369
00:13:36,867 --> 00:13:39,970
DIDN'T HAPPEN IN THIS TRIAL BUT

370
00:13:39,970 --> 00:13:42,773
REALLY WAS NO MUTANT SPLICING

371
00:13:42,773 --> 00:13:43,840
SIGNATURE SO RIGHT, SO NOW THIS

372
00:13:43,840 --> 00:13:45,375
DRUG IS NOT ON THE MARKET,

373
00:13:45,375 --> 00:13:46,476
ALTHOUGH PEOPLE AGAIN CAREFULLY

374
00:13:46,476 --> 00:13:48,445
LOOKING AT THE DATA AND

375
00:13:48,445 --> 00:13:50,180
HOPEFULLY FINDING MAYBE A SUBSET

376
00:13:50,180 --> 00:13:51,481
OR COMBINATIONS OF OTHER

377
00:13:51,481 --> 00:13:52,583
MECHANISMS WHERE IT CAN BE USED

378
00:13:52,583 --> 00:13:54,518
IN THE TREATMENT OF MDECKER S

379
00:13:54,518 --> 00:13:55,586
BECAUSE WE DON'T HAVE ANYTHING

380
00:13:55,586 --> 00:13:59,222
GOOD TO GIVE TO PATIENTS.

381
00:13:59,222 --> 00:14:03,760
SO AT THAT SAME TIME, YOU KNOW

382
00:14:03,760 --> 00:14:06,229
RIGHT, ONCE YOU DEVELOP A REALLY

383
00:14:06,229 --> 00:14:07,431
GOOD APPROACH, ECNOLOGY, IT WAS

384
00:14:07,431 --> 00:14:11,535
OBVIOUS FOR US, OKAY, LET'S

385
00:14:11,535 --> 00:14:12,936
TACKLE U2 FAC1, BECAUSE THAT'S

386
00:14:12,936 --> 00:14:16,273
WHAT WE CAN DO IN THE LAB, WE

387
00:14:16,273 --> 00:14:17,441
DIDN'T WANT TO REINVENT THE

388
00:14:17,441 --> 00:14:20,110
WHILE, WE DIDN'T WANT TO MOVE ON

389
00:14:20,110 --> 00:14:20,677
TO MANAGE NEW.

390
00:14:20,677 --> 00:14:22,980
SO WE LOOKED AT MUTANT 1 BIND

391
00:14:22,980 --> 00:14:24,715
AND HOW DO THEY ALTER BINDING

392
00:14:24,715 --> 00:14:27,284
AND THERE WAS THIS VERY SIMPLE,

393
00:14:27,284 --> 00:14:28,452
YOU KNOW HYPOTHESIS THAT PEOPLE

394
00:14:28,452 --> 00:14:32,289
HAD FROM IN INVITRO DATA AND

395
00:14:32,289 --> 00:14:33,690
ALSO RNA SEQUENCING DATA WHERE

396
00:14:33,690 --> 00:14:36,560
IT SAID, YOU KNOW, SO IT MUST

397
00:14:36,560 --> 00:14:39,162
JUST NOT BIND AND THEN HAVE YOU

398
00:14:39,162 --> 00:14:39,796
EXON EXCLUSION.

399
00:14:39,796 --> 00:14:41,732
I SAID WELL, LET'S LOOK AT OUR

400
00:14:41,732 --> 00:14:44,701
TECHNOLOGY AND WHEN I SAY LET'S

401
00:14:44,701 --> 00:14:46,103
LOOK, I TOLD YULIA CAN YOU

402
00:14:46,103 --> 00:14:48,238
PLEASE LOOK AND SO JULIA IS AN

403
00:14:48,238 --> 00:14:49,272
AMAZING POST DOC, AND JUNIOR

404
00:14:49,272 --> 00:14:50,941
SCIENTIST IN THE LAB ACTUALLY

405
00:14:50,941 --> 00:14:54,478
LOOKING FOR FACULTY POSITION,

406
00:14:54,478 --> 00:14:55,879
SHE'S UTTERLY AMAZING, SHE'S A

407
00:14:55,879 --> 00:14:56,613
TRIPLE THREAT THAT EVERYONE

408
00:14:56,613 --> 00:14:59,783
WANTS TO HAVE IN THE LAB,

409
00:14:59,783 --> 00:15:02,019
BIO-INFORMATICS, AMAZING AT THE

410
00:15:02,019 --> 00:15:03,920
BENCH, FANTAST COLLABORATION AND

411
00:15:03,920 --> 00:15:07,557
TUMOR BIOLOGY WHO FORTUNATELY

412
00:15:07,557 --> 00:15:09,559
WORKED IN MY LAB FOR 3 YEARS AND

413
00:15:09,559 --> 00:15:12,596
I WAS A PROFESSOR IN ITALY SO WE

414
00:15:12,596 --> 00:15:14,331
HAVE JOINT LABS ACROSS THE

415
00:15:14,331 --> 00:15:14,598
ATLANTIC.

416
00:15:14,598 --> 00:15:21,271
SO AGAIN, IF YOU LOOK AT U21 AND

417
00:15:21,271 --> 00:15:22,706
USF2, SO HERE THESE FINGER

418
00:15:22,706 --> 00:15:24,775
DOMAINS THAT HAVE THE POINT

419
00:15:24,775 --> 00:15:27,711
MUTATIONS AND IN THE MIDDLE IS

420
00:15:27,711 --> 00:15:29,112
YOUR END DOMAIN WHERE PEOPLE

421
00:15:29,112 --> 00:15:30,380
HAVE SHOWN BEAUTIFULLY IT'S

422
00:15:30,380 --> 00:15:31,615
THESE 2 ZINC FINGERS THAT

423
00:15:31,615 --> 00:15:33,784
COMBINE AND ARE ACTUALLY THE RNA

424
00:15:33,784 --> 00:15:36,086
BINDING SURFACE WHICH IS

425
00:15:36,086 --> 00:15:36,353
GORGEOUS.

426
00:15:36,353 --> 00:15:38,922
THE PROBLEM IS THAT IT ALWAYS

427
00:15:38,922 --> 00:15:44,094
OCCURS IT'S LIKE AN OBLIGATORY

428
00:15:44,094 --> 00:15:44,394
HETERODIMER.

429
00:15:44,394 --> 00:15:47,931
AND PEOPLE ALWAYS THOUGHT, WELL

430
00:15:47,931 --> 00:15:51,134
WE KNOW U2 AF2 BINDS BUT IT

431
00:15:51,134 --> 00:15:52,803
HADN'T BEEN PROVEN IN VIVO.

432
00:15:52,803 --> 00:15:58,775
SO WE DECIDED, YOU KNOW TO USE

433
00:15:58,775 --> 00:16:01,812
ADVANCED TECHNOLOGY OVER

434
00:16:01,812 --> 00:16:03,346
[INDISCERNIBLE] CLIP ENHANCED

435
00:16:03,346 --> 00:16:09,219
CROSS LINKS AND IMMUNO

436
00:16:09,219 --> 00:16:09,920
PRECIPITATION, AND PULL OUT

437
00:16:09,920 --> 00:16:11,354
THOSE THAT HAVE A FLAG TAG.

438
00:16:11,354 --> 00:16:12,989
AND WE LABEL THESE RNAs THAT

439
00:16:12,989 --> 00:16:16,126
WE PULL OUT WITH P32 AND WHEN WE

440
00:16:16,126 --> 00:16:17,694
SEQUENCE TWINED OF THE END OF

441
00:16:17,694 --> 00:16:21,064
THE READ FROM THE REVERSE

442
00:16:21,064 --> 00:16:22,799
TRANSCRIPTION IS EXACTLY THE

443
00:16:22,799 --> 00:16:23,567
CROSS LINKING SIDE.

444
00:16:23,567 --> 00:16:26,036
AND BECAUSE WE KNEW THAT YOU

445
00:16:26,036 --> 00:16:28,739
KNOW WE HAVE THIS HETERODIMER,

446
00:16:28,739 --> 00:16:31,808
YULIA DECIDED TO CUT THIS

447
00:16:31,808 --> 00:16:34,644
MEMBRANE, THIS IS THE RIA THAT'S

448
00:16:34,644 --> 00:16:36,947
BOUND, LABELED AS P32 TO CUT THE

449
00:16:36,947 --> 00:16:40,016
MEMBRANE JUST UNDERNEATH THE

450
00:16:40,016 --> 00:16:44,621
SIZE OF U2 AF2, WHICH IS 65

451
00:16:44,621 --> 00:16:45,021
KILODALTONS.

452
00:16:45,021 --> 00:16:47,724
AND THEN THE BOUND RNETWORKA

453
00:16:47,724 --> 00:16:49,593
SHIFT IS BOUND THESE PROTEINS

454
00:16:49,593 --> 00:16:49,893
UP.

455
00:16:49,893 --> 00:16:51,928
SO SHE CUT THE MEMBRANE TO

456
00:16:51,928 --> 00:16:55,265
RELOOK AT, OKAY, WHAT DOES U2

457
00:16:55,265 --> 00:16:56,500
AF1 RECEIVE THE DIFFERENCES?

458
00:16:56,500 --> 00:16:57,501
SO UP HERE WE DIDN'T KNOW WHAT

459
00:16:57,501 --> 00:17:01,037
WE WERE GOING TO GET, WERE WE

460
00:17:01,037 --> 00:17:05,041
GETTING U2 AF2 OR

461
00:17:05,041 --> 00:17:05,442
HETEROBINDINGING.

462
00:17:05,442 --> 00:17:08,478
SO HERE YOU SEE THE

463
00:17:08,478 --> 00:17:09,279
[INDISCERNIBLE] TRACK AND YOU

464
00:17:09,279 --> 00:17:11,815
CAN SEE THE VERY BRIGHT CLEAR

465
00:17:11,815 --> 00:17:14,017
PEEK OF U2 F1 BINDING AT THE 3

466
00:17:14,017 --> 00:17:17,521
PRIME SPLICE SIDE AND THEN RIGHT

467
00:17:17,521 --> 00:17:18,822
IN THIS LARGER BAND THAT WE

468
00:17:18,822 --> 00:17:22,459
PULLED OUT, YOU CAN SEE THAT

469
00:17:22,459 --> 00:17:24,995
THERE'S U2 AF1 BINDING AND NOW

470
00:17:24,995 --> 00:17:29,299
YOU SEE U2 AF2 BINDING OVER THE

471
00:17:29,299 --> 00:17:29,533
TRACT.

472
00:17:29,533 --> 00:17:32,202
IF YOU LOOK AT WHAT DO THE

473
00:17:32,202 --> 00:17:36,106
MUTATIONS DO, SO ORANGE THE S34

474
00:17:36,106 --> 00:17:39,743
F-MUTATION IN THE FIRST FINGER

475
00:17:39,743 --> 00:17:41,378
AND THE R-MUTATION THE

476
00:17:41,378 --> 00:17:42,679
SEQUENCING FINGER, YOU CAN SEE

477
00:17:42,679 --> 00:17:43,480
THEY'RE DIFFERENT, THEY'RE NOT

478
00:17:43,480 --> 00:17:45,982
THE SAME SO CAN YOU SEE THAT

479
00:17:45,982 --> 00:17:51,621
WITH F34 F WE GAIN A MINUS 3 P,

480
00:17:51,621 --> 00:17:55,859
OVER THIS C VERSUS U-3 POSITION

481
00:17:55,859 --> 00:17:58,028
AND WITH Q87 WE GAIN A VERY

482
00:17:58,028 --> 00:17:58,795
SUBTLE PLUS 1 PEAK.

483
00:17:58,795 --> 00:18:00,430
AND THEN THE OTHER THING YOU CAN

484
00:18:00,430 --> 00:18:04,267
SEE THAT THE MUTATIONS, THEY NOT

485
00:18:04,267 --> 00:18:06,369
ONLY AFFECT THE U2 F 1 BINDING

486
00:18:06,369 --> 00:18:08,538
BUT MABEL THEY ALSO AFFECT HOW

487
00:18:08,538 --> 00:18:11,842
THE WAVE INTERACTING WITH THE R,

488
00:18:11,842 --> 00:18:14,411
INA AND THEN IF WE ARE NOT

489
00:18:14,411 --> 00:18:15,946
COMBINE THIS WITH THE SEQUENCING

490
00:18:15,946 --> 00:18:18,381
DATA AND THAT HAD ALREADY BEEN

491
00:18:18,381 --> 00:18:21,017
PUB LESHED BEFORE, IS THERE IN

492
00:18:21,017 --> 00:18:23,753
BINDING WE ACTUALLY DO NOT HAVE

493
00:18:23,753 --> 00:18:29,326
A SEQUENCE SPECIFICITY TO

494
00:18:29,326 --> 00:18:31,194
WHETHER YOU--S34 F-MUTANT BINDS

495
00:18:31,194 --> 00:18:33,530
OR DOESN'T BIND, IT BINDS

496
00:18:33,530 --> 00:18:34,698
EQUALLY WELL, IT DOESN'T MATTER

497
00:18:34,698 --> 00:18:37,000
WHAT THE SEQUENCE IS AT THE 3,

498
00:18:37,000 --> 00:18:38,635
OR WHAT THE SEQUENCE IS AT THE +

499
00:18:38,635 --> 00:18:40,837
1 BUT WHEN YOU LOOK AT ACTUAL

500
00:18:40,837 --> 00:18:42,806
SPLICING, IF YOU HAVE U AT THAT

501
00:18:42,806 --> 00:18:45,575
MINUS 3 POSITION, THE EXON IS

502
00:18:45,575 --> 00:18:46,676
ACTUALLY PREVENTIVE

503
00:18:46,676 --> 00:18:47,544
REPRESENTIALLY EXCLUDED IF YOU

504
00:18:47,544 --> 00:18:55,318
HAVE A C IT'S PREFERENTIALLY

505
00:18:55,318 --> 00:18:58,321
INCLUDED. 

506
00:18:58,321 --> 00:18:59,990
SO SPLICING DOESN'T NECESSARILY

507
00:18:59,990 --> 00:19:01,758
MEAN SPLICING IN THE OUTCOME SO

508
00:19:01,758 --> 00:19:03,159
A FEW MORE WORK ON DO.

509
00:19:03,159 --> 00:19:04,895
SO THIS IS NICE AND IT'S NOT

510
00:19:04,895 --> 00:19:08,365
LIKE ALL OR NOTHING OR BLACK AND

511
00:19:08,365 --> 00:19:08,765
WHITE.

512
00:19:08,765 --> 00:19:10,767
SO JULIA REALLY INTEGRATED

513
00:19:10,767 --> 00:19:13,403
BINDING AND SPLICING AND IT

514
00:19:13,403 --> 00:19:15,739
SEEMS THAT U2 AF1 MUTATIONS LOOK

515
00:19:15,739 --> 00:19:17,607
LIKE MORE OF GAIN OF BYPASSING,

516
00:19:17,607 --> 00:19:20,543
LOTS OF SPLICING MODELS, THAT'S

517
00:19:20,543 --> 00:19:22,245
THIS QUADRANT HERE, MORE BINDING

518
00:19:22,245 --> 00:19:23,580
BUT THERE'S LESS EXIT AND

519
00:19:23,580 --> 00:19:25,448
INCLUSION AND THIS IS AN EXAMPLE

520
00:19:25,448 --> 00:19:28,184
OF BOTH, YOU KNOW THE U2 F 1 AND

521
00:19:28,184 --> 00:19:29,386
THEN THE COMBINED BINDING AND

522
00:19:29,386 --> 00:19:39,596
YOU CAN SEE THAT THERE'S A

523
00:19:39,596 --> 00:19:41,197
REDUCED BINDING THAT IS NEEDED

524
00:19:41,197 --> 00:19:42,399
FOR THE INCLUSION OF THE EXON

525
00:19:42,399 --> 00:19:47,203
AND THEN WHEN YOU LOOK AT U2 AF1

526
00:19:47,203 --> 00:19:48,338
MUTATIONS, THAT LOOKS MORE LIKE

527
00:19:48,338 --> 00:19:50,774
A LOSS OF BINDING, LOSS OF

528
00:19:50,774 --> 00:19:52,309
SPLICING MODEL KIND OF SHOWN

529
00:19:52,309 --> 00:19:55,145
HERE, RIGHT, SO THERE'S LESS OF

530
00:19:55,145 --> 00:19:57,981
A PEAK AT THIS 1 POSITION AND

531
00:19:57,981 --> 00:20:02,719
ALSO YOU AGAIN HAVE KIND OF LESS

532
00:20:02,719 --> 00:20:04,421
BINDING OF U2 F2, BUT OF COURSE

533
00:20:04,421 --> 00:20:06,456
YOU HAVE DOTS IN ALL THE

534
00:20:06,456 --> 00:20:06,957
QUADRANTS.

535
00:20:06,957 --> 00:20:08,825
SO IN THE NEXT STEP, YULIA

536
00:20:08,825 --> 00:20:10,760
LOOKED AT OKAY, WE WANTED TO

537
00:20:10,760 --> 00:20:15,699
UNDERSTAND WHAT ARE THE DIRECT

538
00:20:15,699 --> 00:20:19,169
EFFECTS OF THESE MUTATIONS SO WE

539
00:20:19,169 --> 00:20:20,003
START BY INTEGRATING, BINDING

540
00:20:20,003 --> 00:20:22,372
AND SPLICING AND LOOKING AT

541
00:20:22,372 --> 00:20:24,574
PATHWAYS THAT WE WOULDN'T LOOK

542
00:20:24,574 --> 00:20:25,842
AT SECONDARY EFFECTS BECAUSE ANY

543
00:20:25,842 --> 00:20:27,510
EFFECT BY THESE CELLS GROW FOR

544
00:20:27,510 --> 00:20:29,245
SOMETIME, SO IF YOU JUST LOOK AT

545
00:20:29,245 --> 00:20:30,780
PLIESING, THERE MAY BE MANY

546
00:20:30,780 --> 00:20:32,649
SECONDARY EFFECTS AND IN

547
00:20:32,649 --> 00:20:34,150
PARTICULAR, WE SEE DIFIENTIAL

548
00:20:34,150 --> 00:20:36,419
SPLICING OF OTHER RNA BINDING

549
00:20:36,419 --> 00:20:37,787
PROTEINS, WELL THERE'S KIND OF

550
00:20:37,787 --> 00:20:39,456
THIS CASCADE, BUT BY LOOKING

551
00:20:39,456 --> 00:20:42,625
DREBLGHTLY AT BINDING AND

552
00:20:42,625 --> 00:20:44,160
SPLICING, YOU KNOW MAYBE ABLE TO

553
00:20:44,160 --> 00:20:45,428
LOOK AT DIRECT EFFECTS, AND WHAT

554
00:20:45,428 --> 00:20:48,732
YOU FOUND IN AN ENRICHMENT FOR

555
00:20:48,732 --> 00:20:53,703
THIS, REGULATION OF ORGANELLE

556
00:20:53,703 --> 00:20:56,573
ASSEMBLY, OR CYTOPLASMIC STRESS

557
00:20:56,573 --> 00:20:58,208
GRANULES AND BEFORE THIS THREAD,

558
00:20:58,208 --> 00:20:59,843
MANY OTHER PATHWAYS WERE

559
00:20:59,843 --> 00:21:01,945
IDENTIFIED BUT IN OUR DATA,

560
00:21:01,945 --> 00:21:04,914
BYPASSING, SPLICING, THIS IS A

561
00:21:04,914 --> 00:21:06,549
REALLY PREDOMINANT NODE STRESS

562
00:21:06,549 --> 00:21:08,151
GRANULES BUT ALSO CALLED

563
00:21:08,151 --> 00:21:10,820
[INDISCERNIBLE] BODIES AND OTHER

564
00:21:10,820 --> 00:21:12,155
NUCLEAR BODIES, ASSEMBLIES.

565
00:21:12,155 --> 00:21:15,158
SO, WHAT OUR STRESS GRANULES, I

566
00:21:15,158 --> 00:21:17,394
GREAN THINK IT'S TOTALLY

567
00:21:17,394 --> 00:21:18,895
BEAUTIFUL SCIENCE AND I'M VERY

568
00:21:18,895 --> 00:21:20,663
TBLAD THAT JULIA AS A PUNISHING

569
00:21:20,663 --> 00:21:22,432
HD IS KIND OF GOING INTO THAT

570
00:21:22,432 --> 00:21:24,401
FEE AND I'M LOOKING AT IT LIKE

571
00:21:24,401 --> 00:21:26,669
WOW, THIS IS TOO COMPLICATED FOR

572
00:21:26,669 --> 00:21:29,105
ME, BUT STRESS GRANULES AND MANY

573
00:21:29,105 --> 00:21:30,740
OTHER GRANULES, THEY ARE

574
00:21:30,740 --> 00:21:32,675
ORGANELLES AND THEY RIGHT, SO

575
00:21:32,675 --> 00:21:36,980
THEY'RE LIKE PHYSICAL CHEMICAL

576
00:21:36,980 --> 00:21:37,914
TIMING, WHATEVER, CONGRAMERATES

577
00:21:37,914 --> 00:21:39,249
OF CERTAIN PROTEINS AND RNAs

578
00:21:39,249 --> 00:21:40,917
AND I THINK THE BEAUTY AND

579
00:21:40,917 --> 00:21:45,722
STRESS GRANULES IS THAT THEY CAN

580
00:21:45,722 --> 00:21:46,289
BE--PROBABLY--IT'S PROBABLY

581
00:21:46,289 --> 00:21:48,825
ENERGY EFFICIENT TO REGULATE

582
00:21:48,825 --> 00:21:49,459
TRANSLATION AND CELLULAR

583
00:21:49,459 --> 00:21:51,428
FUNCTION THAT WAY AND IT'S

584
00:21:51,428 --> 00:21:53,063
PROBABLY ALSO, IT CAN HAPPEN

585
00:21:53,063 --> 00:21:55,131
FAST BECAUSE YOU DON'T HAVE THIS

586
00:21:55,131 --> 00:22:01,071
ACTIVE TRANSPORT ACROSS

587
00:22:01,071 --> 00:22:01,738
MEMBRANES.

588
00:22:01,738 --> 00:22:03,006
AND [INDISCERNIBLE] SMART, SO

589
00:22:03,006 --> 00:22:05,075
PEOPLE HAVE SHOWN THAT STRESS

590
00:22:05,075 --> 00:22:07,510
GRANULES SEEM TO PROVIDE AN

591
00:22:07,510 --> 00:22:09,279
ADVANTAGE TO CANCERS AND SO

592
00:22:09,279 --> 00:22:10,447
COULD STRESS GRANULES BE A

593
00:22:10,447 --> 00:22:14,384
MECHANISM BY WHICH THESE MUTANT

594
00:22:14,384 --> 00:22:17,387
CELLS GAIN THAT ADVANTAGE IN

595
00:22:17,387 --> 00:22:19,422
ELDERLY PATIENTS BONE MARROW

596
00:22:19,422 --> 00:22:20,990
OVERTIME SO THE FIRST THING SHE

597
00:22:20,990 --> 00:22:23,293
HAD TO SHOW THAT YES THERE ARE

598
00:22:23,293 --> 00:22:26,863
MORE STRESS GRANULES SO THIS IS

599
00:22:26,863 --> 00:22:30,400
INDUCIBLE CELL LINES, AND TO USE

600
00:22:30,400 --> 00:22:31,101
1 STRESSOXIDATIVE STRESS WITH

601
00:22:31,101 --> 00:22:32,735
[INDISCERNIBLE] AND YOU CAN SEE

602
00:22:32,735 --> 00:22:33,903
THERE ARE MORE STRESS GRANULES

603
00:22:33,903 --> 00:22:35,805
IN THE MUTANT CELLS AND THE

604
00:22:35,805 --> 00:22:37,774
WILD-TYPE CELLS, THESE CELL

605
00:22:37,774 --> 00:22:39,976
LINES WEOXON NOWSLY EXPRESS, YOU

606
00:22:39,976 --> 00:22:43,179
HAVE A MUTANT F1 AND YOU TOOK

607
00:22:43,179 --> 00:22:45,148
PATIENT SAMPLES FROM PATIENTS

608
00:22:45,148 --> 00:22:48,218
WITH SECONDARY AML FROM MDS WHO

609
00:22:48,218 --> 00:22:50,854
ARE EITHER WILD-TYPE OR MUTANT

610
00:22:50,854 --> 00:22:53,156
FOR UGF 1 BUT FOR OTHER SPLICING

611
00:22:53,156 --> 00:22:57,494
FACTORS AND SEE CAN SEE STRESS

612
00:22:57,494 --> 00:22:58,428
GRANULES.

613
00:22:58,428 --> 00:23:00,697
SO, IN ANOTHER--BECAUSE AGAIN 1

614
00:23:00,697 --> 00:23:02,999
CAN SEE LONG INVITRO OF THE CELL

615
00:23:02,999 --> 00:23:04,367
LINES, ET CETERA BUT SHE ALSO

616
00:23:04,367 --> 00:23:06,903
WANTED TO KNOW OKAY, CAN SHE

617
00:23:06,903 --> 00:23:08,438
FIGURE OUT WHAT THESE MUTATIONS

618
00:23:08,438 --> 00:23:11,708
DO, IN THE SINGLE CELL LEVEL IN

619
00:23:11,708 --> 00:23:13,309
THE HEMATOPOIETIC HIERARCHY AND

620
00:23:13,309 --> 00:23:18,281
SHE DID 10 X SEQUENCING ON 5 MDS

621
00:23:18,281 --> 00:23:19,749
PEASHTS WITHOUT ANY SPECIFIC

622
00:23:19,749 --> 00:23:22,051
MUTATIONS AND 3 MDS PASHTS WITH

623
00:23:22,051 --> 00:23:24,521
[INDISCERNIBLE] F MUTATIONS AND

624
00:23:24,521 --> 00:23:27,056
SHE ACTUALLY USED 10 X 5 PRIME

625
00:23:27,056 --> 00:23:30,460
LIBRARY, WHICH IS NICE BECAUSE

626
00:23:30,460 --> 00:23:32,996
THIS S34 F MUTATION, RIGHT, IS

627
00:23:32,996 --> 00:23:34,397
VERY, VERY 5 PRIME AND WE'RE

628
00:23:34,397 --> 00:23:37,033
HOPING THAT MAYBE EVEN IN THE

629
00:23:37,033 --> 00:23:38,768
SINGLE CELL, SCHWARTZ THAT THEY

630
00:23:38,768 --> 00:23:40,803
WOULD GET, THAT YOU WOULD BE

631
00:23:40,803 --> 00:23:43,139
ABLE TO SEE WIDER VERSUS MUTANT

632
00:23:43,139 --> 00:23:46,676
READS SO SHE CAN DO THAT RIGHT,

633
00:23:46,676 --> 00:23:48,878
SO THE IN GRAY, ARE THE CELLS

634
00:23:48,878 --> 00:23:52,582
FROM PATIENTS WHO DON'T HAVE

635
00:23:52,582 --> 00:23:53,583
MUTATIONS, IN THIS DARK ORANGE

636
00:23:53,583 --> 00:23:55,718
ARE CELLS FROM PATIENTS WITH S34

637
00:23:55,718 --> 00:23:56,886
FMARKED FOR IDENTIFICATION

638
00:23:56,886 --> 00:23:57,954
MUTATIONS WHERE FOR EACH CELL

639
00:23:57,954 --> 00:24:00,790
SHE COULD SEE AT LEAST 1 MUTANT

640
00:24:00,790 --> 00:24:03,326
READ AND THEN SLIDE ORANGE ARE

641
00:24:03,326 --> 00:24:05,295
CELLS WHERE THERE WERE TO HAVE

642
00:24:05,295 --> 00:24:06,596
READS BUT NONE OF THEM MUTANT

643
00:24:06,596 --> 00:24:08,798
BUT IF YOU DON'T HAVE ENOUGH

644
00:24:08,798 --> 00:24:09,966
READS, YOU CAN'T NECESSARILY

645
00:24:09,966 --> 00:24:12,869
EXCLUDE THAT THEY'RE MUTANT AND

646
00:24:12,869 --> 00:24:17,807
SORRY, I'LL KEEP CLICKING HERE

647
00:24:17,807 --> 00:24:20,877
AND THEN, RIGHT FROM HER BINDING

648
00:24:20,877 --> 00:24:21,744
SPLICING [INDISCERNIBLE], AND

649
00:24:21,744 --> 00:24:24,514
THEN PUBLISH DATA SETS

650
00:24:24,514 --> 00:24:27,917
PROTEOMICS AND DATA SETS ON ICE

651
00:24:27,917 --> 00:24:30,687
GRANULES, SHE DESCRIBED A STRESS

652
00:24:30,687 --> 00:24:32,188
GRANULE SCORE AND SHE CAN APPLY

653
00:24:32,188 --> 00:24:34,157
THIS TO THE SINGLE CELLS.

654
00:24:34,157 --> 00:24:35,592
I THINK I YUSMED A SLIGHT SO I

655
00:24:35,592 --> 00:24:36,893
THINK I WILL GO BACK TO THAT SO

656
00:24:36,893 --> 00:24:41,898
YOU CAN SEE THAT THE CELLS WITH

657
00:24:41,898 --> 00:24:43,866
MUTANT READINGS REALLY HAVE A

658
00:24:43,866 --> 00:24:45,335
HIGH STRESS GRANULE SCORE

659
00:24:45,335 --> 00:24:46,436
COMPARED TO WILD-TYPE CELLS AND

660
00:24:46,436 --> 00:24:49,405
THESE CELLS WHERE WE DON'T KNOW

661
00:24:49,405 --> 00:24:50,440
OTHER WILD-TYPE MUTANT THEY FALL

662
00:24:50,440 --> 00:24:52,008
IN BETWEEN AND SORRY I JUMPED

663
00:24:52,008 --> 00:24:54,410
HERE, AND WHAT YOU SEE HERE NOW

664
00:24:54,410 --> 00:24:57,180
IS RIGHT, SO THIS REGION HERE

665
00:24:57,180 --> 00:25:00,550
HAS THE PROGENITOR CELLS, GRANUE

666
00:25:00,550 --> 00:25:02,852
LOW CITES, MONOCYTES AND THE

667
00:25:02,852 --> 00:25:08,157
MUTANT CELLS SEEM TO BE ARRESTED

668
00:25:08,157 --> 00:25:10,627
IN THAT MORE PROGENITOR CELL,

669
00:25:10,627 --> 00:25:14,831
AND IF ANYTHING THEY SEEM TO

670
00:25:14,831 --> 00:25:16,466
HAVE MORE MONOSITTIC AND WE SEE

671
00:25:16,466 --> 00:25:20,169
THIS IN PATIENTS WITH ANEMIA AS

672
00:25:20,169 --> 00:25:21,104
IT FITS THEIR PHENOTYPE.

673
00:25:21,104 --> 00:25:22,538
AS A SUMMARY OF THE FIRST PART,

674
00:25:22,538 --> 00:25:26,676
RIGHT, SO IT SEEMS THAT SPLICE

675
00:25:26,676 --> 00:25:27,310
MUTATIONS, USUALLY 1 MUTATIONS

676
00:25:27,310 --> 00:25:28,911
RESULT IN THE STRESS GRANULE

677
00:25:28,911 --> 00:25:32,882
RESPONSE AND IS THAT MAYBE A

678
00:25:32,882 --> 00:25:33,950
MECHANISM THAT ALLOWS THESE

679
00:25:33,950 --> 00:25:36,919
CELLS TO SURVIVE IN AN ADVERSE

680
00:25:36,919 --> 00:25:37,887
ENVIRONMENT WITH AGING AND THEN

681
00:25:37,887 --> 00:25:42,592
THE OTHER QUESTION IS, WAS IT

682
00:25:42,592 --> 00:25:43,559
COMMON TO OTHER LIMITATIONS,

683
00:25:43,559 --> 00:25:45,428
TOO, MAYBE ARE THEY TARGETABLE

684
00:25:45,428 --> 00:25:47,163
IN IS THIS PROCESS TARGETABLE IN

685
00:25:47,163 --> 00:25:52,168
SO THEN WE WENT BACK TO OUR SSF2

686
00:25:52,168 --> 00:25:53,469
MUTANT CELL ANDS DATA AND I

687
00:25:53,469 --> 00:25:55,471
DIDN'T SHOW THIS EARLIER BUT

688
00:25:55,471 --> 00:25:58,875
WHEN WE INITIALLY DID OUR RNA

689
00:25:58,875 --> 00:26:00,543
SEQUENCING, ANALYSIS, WE WERE A

690
00:26:00,543 --> 00:26:02,779
LITTLE BIT AT A LOSS BECAUSE WE

691
00:26:02,779 --> 00:26:03,980
SAW THE DIFFERENTIAL SPLICING OF

692
00:26:03,980 --> 00:26:05,748
MANY OTHER R, INA BINDING

693
00:26:05,748 --> 00:26:07,950
PROTEINS, MANY OF THESE ARE

694
00:26:07,950 --> 00:26:10,353
INVOLVED IN YOU KNOW IN STRESS

695
00:26:10,353 --> 00:26:13,690
GRANULES, FOR EXAMPLE, AND OTHER

696
00:26:13,690 --> 00:26:22,665
MEMBRANES ORGANELLES, AND THEN,

697
00:26:22,665 --> 00:26:25,501
JULIA ANALYZED ALL THE SRF 2

698
00:26:25,501 --> 00:26:27,036
MUTANT PLIESING CELL LINE AT

699
00:26:27,036 --> 00:26:29,105
PATIENT DATA SETS AND ALSO FOUND

700
00:26:29,105 --> 00:26:31,574
AGAIN SOME ENRICHMENT IN THIS

701
00:26:31,574 --> 00:26:32,809
STRESS GRANULE SIGNATURE, AND

702
00:26:32,809 --> 00:26:36,813
THEN AGAIN SHE SEQUENCED--USED

703
00:26:36,813 --> 00:26:39,048
THE SAME PATIENTS BUT ALSO

704
00:26:39,048 --> 00:26:40,850
SEQUENCE SF 2 MUTANT PATIENTS

705
00:26:40,850 --> 00:26:43,686
AGAIN WITH THE 3 BIEM

706
00:26:43,686 --> 00:26:45,988
SEQUENCING, SO RIGHT S34 F IS

707
00:26:45,988 --> 00:26:48,658
THE ESEN 34, P95 AND AN AMINO

708
00:26:48,658 --> 00:26:51,761
ACID, 95, SO MUCH, MUCH, HARDER

709
00:26:51,761 --> 00:26:54,197
TO ACTUALLY FIND READS THAT GO

710
00:26:54,197 --> 00:26:56,265
THAT FAR INTO, YOU KNOW INTO THE

711
00:26:56,265 --> 00:26:56,733
SEQUENCE.

712
00:26:56,733 --> 00:26:57,900
BUT EVEN THERE, THERE'S A

713
00:26:57,900 --> 00:26:59,769
SUGWREOF THE YEN THAT THERE MAY

714
00:26:59,769 --> 00:27:02,238
BE AN INCREASED STRESS RESPONSE

715
00:27:02,238 --> 00:27:06,109
SO THAT COULD BE A COMMON

716
00:27:06,109 --> 00:27:06,509
MECHANISM.

717
00:27:06,509 --> 00:27:09,512
SO THEN THE QUESTION IS HEY,

718
00:27:09,512 --> 00:27:14,050
DOES IT HAVE ANY BIOLOGIC

719
00:27:14,050 --> 00:27:15,785
FUNCTION AND ONCE SHE TAKES THE

720
00:27:15,785 --> 00:27:17,620
WILD-TYPE OR THE CELLS AND

721
00:27:17,620 --> 00:27:18,354
EXPOSES THEM TO

722
00:27:18,354 --> 00:27:19,389
ARE--ADMINISTRATIVE SONNITE AND

723
00:27:19,389 --> 00:27:20,923
LOOKS AT CELL VIABILITY, THERE

724
00:27:20,923 --> 00:27:24,026
SEEMS TO BE SOME ADVANTAGE IN

725
00:27:24,026 --> 00:27:24,694
THE MUTANT CELLS.

726
00:27:24,694 --> 00:27:27,864
AND THERE IS AN INHIBITOR OF

727
00:27:27,864 --> 00:27:32,468
THIS INTEGRATED TRESES RESPONSE

728
00:27:32,468 --> 00:27:34,237
CALLED ISRIP AND IT FUNCTIONS

729
00:27:34,237 --> 00:27:37,140
HERE, IT INHIBITS THE FUNCTION

730
00:27:37,140 --> 00:27:38,074
OF PHOSPHORYLATED EIF2 ALPHA

731
00:27:38,074 --> 00:27:40,543
WHICH IS IN THE STRESS RESPONSE

732
00:27:40,543 --> 00:27:43,546
THAT WOULD LEAD TO STOP OF

733
00:27:43,546 --> 00:27:43,913
TRANSLATION.

734
00:27:43,913 --> 00:27:46,582
AND I LEARNED READING ABOUT

735
00:27:46,582 --> 00:27:49,352
ISRIB, THAT I THINK IT'S BEEN

736
00:27:49,352 --> 00:27:51,187
USED, YOU KNOW PEOPLE WHO WANT

737
00:27:51,187 --> 00:27:52,722
TO NOT AGE OR ANYTHING LIKE THAT

738
00:27:52,722 --> 00:27:55,792
BUT ALSO IN ALL SORTS OF THINGS.

739
00:27:55,792 --> 00:27:57,994
SO WE DON'T KNOW IF THIS IS

740
00:27:57,994 --> 00:27:59,829
VIABLE DRUG OR NOT THAT PEOPLE

741
00:27:59,829 --> 00:28:01,798
COULD TAKE WITH THESE DISEASES,

742
00:28:01,798 --> 00:28:03,566
BUT HE TESTED IT INVITRO IN OUR

743
00:28:03,566 --> 00:28:06,002
CELL LINES AND THE EFFECT IS

744
00:28:06,002 --> 00:28:08,204
VERY, VERY SUBTLE, HENS THIS

745
00:28:08,204 --> 00:28:09,238
REPRESENTATION BUT THERE SEEMS

746
00:28:09,238 --> 00:28:12,008
TO BE A DIFFERENTIAL EFFECT OF

747
00:28:12,008 --> 00:28:14,477
THIS INHIBITOR ON MUTANT

748
00:28:14,477 --> 00:28:15,077
COMPARED TO WILD-TYPE CELLS.

749
00:28:15,077 --> 00:28:18,381
SO THIS IS WHAT SHE HAD SO FAR.

750
00:28:18,381 --> 00:28:20,550
AND THEN OBVIOUSLY, THE OBVIOUS

751
00:28:20,550 --> 00:28:21,751
QUESTION AND THOMAS ASKED ME,

752
00:28:21,751 --> 00:28:24,353
WELL IS IT DIRECT, IS IT

753
00:28:24,353 --> 00:28:25,822
INCORRECT AND WE DON'T KNOW.

754
00:28:25,822 --> 00:28:27,256
BUT THE FIRST PLACE TO ELECTRIC

755
00:28:27,256 --> 00:28:34,297
NOW IS OKAY, WHAT IS IN THESE

756
00:28:34,297 --> 00:28:34,664
STRESS GRANULES?

757
00:28:34,664 --> 00:28:36,833
AND I THINK IT'S DIFFICULT TO

758
00:28:36,833 --> 00:28:42,872
ISOLATE LITTLE  ORGANELLES IN TE

759
00:28:42,872 --> 00:28:44,474
MEMBRANES, HOW DO YOU ISOLATE

760
00:28:44,474 --> 00:28:47,376
THEM AND ARE THIS PHYSICAL

761
00:28:47,376 --> 00:28:49,745
CONGLOMERATE BUT SHE GOT A

762
00:28:49,745 --> 00:28:51,113
PROTOCOL FROM [INDISCERNIBLE]

763
00:28:51,113 --> 00:28:52,315
LAB WHO'S THE WORLD EXPERT IN

764
00:28:52,315 --> 00:28:54,984
THIS FIELD AND IT'S A VERY

765
00:28:54,984 --> 00:28:59,355
COMPLICATED PROCESS WITH LOTS OF

766
00:28:59,355 --> 00:29:00,256
SENTRIFICATIONS, LABELING OF

767
00:29:00,256 --> 00:29:01,123
THESE STRESS GRANULES AND THEN

768
00:29:01,123 --> 00:29:03,759
YOU CAN PULL THEM OUT WITH 1 OF

769
00:29:03,759 --> 00:29:08,798
THE STRESS GRANULE, KIND OF LIKE

770
00:29:08,798 --> 00:29:10,867
MARKET PROTEINS GPP31, SO SHE

771
00:29:10,867 --> 00:29:14,170
DID THIS ON CELLS WITH OR

772
00:29:14,170 --> 00:29:16,105
WITHOUT ARSONITE STRESS, AND

773
00:29:16,105 --> 00:29:19,642
SUBMITTED INPUT AND THE IP

774
00:29:19,642 --> 00:29:21,878
PRODUCT BOTH MASS SPEC AND

775
00:29:21,878 --> 00:29:23,946
RNA-SEQ AND THIS IS RECENT DATA

776
00:29:23,946 --> 00:29:25,381
AND WE STILL DOESN'T HAVE AN

777
00:29:25,381 --> 00:29:25,615
ANSWER.

778
00:29:25,615 --> 00:29:29,986
BUT SHE WAS ABLE TO YOU KNOW

779
00:29:29,986 --> 00:29:37,093
PULL OUT 37 DIFFERENTIALLY 37

780
00:29:37,093 --> 00:29:40,930
PROTEINS, THAT WERE PRESENT IN

781
00:29:40,930 --> 00:29:45,768
S34 F MUTANT CELLS, TRESES

782
00:29:45,768 --> 00:29:46,335
GRANULES, VERSUS WILD-TYPE.

783
00:29:46,335 --> 00:29:48,771
AND THEY WERE ALSO ENRICHED OVER

784
00:29:48,771 --> 00:29:50,139
JUST INPUT.

785
00:29:50,139 --> 00:29:53,643
SO THAT'S THE FIRST START AND

786
00:29:53,643 --> 00:29:58,180
THEN, SAME ANALYZING THESE

787
00:29:58,180 --> 00:29:59,582
RNAs, SO THEY'RE--SHE HAS

788
00:29:59,582 --> 00:30:00,917
DIFFERENTIAL GENE EXPRESSION

789
00:30:00,917 --> 00:30:02,318
WITH RNAs ENRICHED IN THE

790
00:30:02,318 --> 00:30:04,353
STRESS GRANULES THAT ARE

791
00:30:04,353 --> 00:30:05,555
DIFFERENTIALLY EXPRESSED IN THE

792
00:30:05,555 --> 00:30:07,757
WILD-TYPE AND THEN ENRICHED IN

793
00:30:07,757 --> 00:30:09,058
THE GRANULES OVER TOTAL SIDE OF

794
00:30:09,058 --> 00:30:13,396
RTHIS,A IN THE CELL, AND SHE

795
00:30:13,396 --> 00:30:14,463
ALSO HAS SPLICING DIFFERENCES,

796
00:30:14,463 --> 00:30:16,132
SO NOW I THINK FOR HER, THIS IS

797
00:30:16,132 --> 00:30:18,601
A PROJECT SHE WILL TAKE ON IN

798
00:30:18,601 --> 00:30:20,369
HER OWN LAB WILL COME TO NOW

799
00:30:20,369 --> 00:30:23,205
FIGURE OUT WHAT THEY ARE AND I

800
00:30:23,205 --> 00:30:25,174
THINK DOES EVERYBODY HAVE A

801
00:30:25,174 --> 00:30:28,144
COLLABORATION HERE WITH THE NCI.

802
00:30:28,144 --> 00:30:30,212
BECAUSE NOW I THINK THE NEXT

803
00:30:30,212 --> 00:30:32,315
WORK WILL BE TO FIND THESE

804
00:30:32,315 --> 00:30:35,418
PROTEINS RNAs AND THEN ALTER

805
00:30:35,418 --> 00:30:36,586
THEIR EXON INCLUSION, EXCLUSION

806
00:30:36,586 --> 00:30:40,156
TO SEE DOES IT ALTER STRESS

807
00:30:40,156 --> 00:30:41,857
GRANULE BIOLOGY.

808
00:30:41,857 --> 00:30:49,365
AND AGAIN, WHAT'S EXCITING I

809
00:30:49,365 --> 00:30:51,567
THINK, MKS, AML, ARE ALL SOLID

810
00:30:51,567 --> 00:30:54,670
TUMORS AND THEY MAY USE SIMILAR

811
00:30:54,670 --> 00:30:57,607
MECHANISMS IN A SIMILAR WAY AND

812
00:30:57,607 --> 00:30:59,241
P-GRANULES THEY'RE INVOLVED IN

813
00:30:59,241 --> 00:31:01,110
MANY CANCER HALLMARKS, SO

814
00:31:01,110 --> 00:31:02,945
HOPEFULFULLY WE WILL BE ABLE TO

815
00:31:02,945 --> 00:31:05,147
FIND A MECHANISM THAT MAYBE

816
00:31:05,147 --> 00:31:06,782
ULTIMATELY NOT OHM LEAD TO

817
00:31:06,782 --> 00:31:09,085
TREATMENT FOR MDS, BUT ALSO

818
00:31:09,085 --> 00:31:14,156
OTHER LEUKEMIAS, AND SO, I'M

819
00:31:14,156 --> 00:31:17,193
SWITCHING NOW, BECAUSE THIS IS

820
00:31:17,193 --> 00:31:18,961
VERY, VERY, RNA HEAVY AND SANDY

821
00:31:18,961 --> 00:31:21,731
INVITED ME BUT I ALSO SAID I'M A

822
00:31:21,731 --> 00:31:22,898
CLINICIAN, LET ME MAYBE SHOW

823
00:31:22,898 --> 00:31:24,433
SOMETHING ELSE THAT WE'RE DOING

824
00:31:24,433 --> 00:31:30,873
IN THE LAB.

825
00:31:30,873 --> 00:31:34,310
SO, WHEN I WAS A POST DOC AND I

826
00:31:34,310 --> 00:31:36,278
WANTED TO STUDY IN SANDY'S LAB,

827
00:31:36,278 --> 00:31:38,848
I HAD 2 PROBLEMS, I HAD NO

828
00:31:38,848 --> 00:31:39,115
SAMPLES.

829
00:31:39,115 --> 00:31:42,551
THERE WERE NO SAMPLES FOR MILE

830
00:31:42,551 --> 00:31:43,019
MYELODYSPLASIA.

831
00:31:43,019 --> 00:31:45,721
AND AT THAT POINT THERE WERE NOT

832
00:31:45,721 --> 00:31:49,025
CONVINCING MOUSE MODELS, BUT I'M

833
00:31:49,025 --> 00:31:52,128
A CLINICIAN, AND SO I ATTENDED

834
00:31:52,128 --> 00:31:54,764
IN-PEASHT SERVICE AND OF COURSE

835
00:31:54,764 --> 00:31:56,165
THAT'S ALWAYS THE WORST, BECAUSE

836
00:31:56,165 --> 00:31:57,533
THE PEOPLE WHO DO WELL ARE NOT

837
00:31:57,533 --> 00:31:59,168
IN THE HOSPITAL AND EVERY TIME I

838
00:31:59,168 --> 00:32:02,104
COME OFF SERVICE, I'M LIKE WHAT

839
00:32:02,104 --> 00:32:02,672
CAN WE DO?

840
00:32:02,672 --> 00:32:05,541
SO I REALLY WANTED TO DEVELOP A

841
00:32:05,541 --> 00:32:07,109
MODEL TO STUDY MDS AND AT THE

842
00:32:07,109 --> 00:32:08,944
TIME THAT I KIND OF WAS A POST

843
00:32:08,944 --> 00:32:12,915
DOC TO THE LAB, WAS VERY

844
00:32:12,915 --> 00:32:17,186
FORTUNATE THAT IN A RICHARD

845
00:32:17,186 --> 00:32:18,688
FLAVELL, WAS DEVELOPING WITH

846
00:32:18,688 --> 00:32:19,522
ANTHONY [INDISCERNIBLE] A POST

847
00:32:19,522 --> 00:32:24,293
DOC IN HIS LAB, THESE MICE WHERE

848
00:32:24,293 --> 00:32:26,595
THEY WANTED TO HAVE A HUMANIZED

849
00:32:26,595 --> 00:32:28,564
IMMUNE SIZE TO STUDY HIV AND

850
00:32:28,564 --> 00:32:30,900
THEN HUMAN RAT CELLS TO STUDY

851
00:32:30,900 --> 00:32:33,836
MALARIA BECAUSE IT WAS IN PART

852
00:32:33,836 --> 00:32:35,371
FUNDED BY THE GATES FOUNDATION

853
00:32:35,371 --> 00:32:37,339
TO IMPROVE GLOBAL HEALTH AND FOR

854
00:32:37,339 --> 00:32:40,076
THAT, THEY DECIDED TO HUMANIZE

855
00:32:40,076 --> 00:32:40,943
THE BONE MARROW BECAUSE THEY

856
00:32:40,943 --> 00:32:45,414
NEEDED TO BE ABLE TO INGRAFT

857
00:32:45,414 --> 00:32:46,682
HEMATOPOIETIC STEM CELLS AND

858
00:32:46,682 --> 00:32:48,451
ALREADY, WE KNEW AT THAT POINT

859
00:32:48,451 --> 00:32:53,456
THAT MDS, YOU CAN TAKE AN A.

860
00:32:53,456 --> 00:32:56,358
L. L. OR MANY A. M. L.S AND PUT

861
00:32:56,358 --> 00:32:57,993
THEM IN THE WORSE DEFIC MOUSE

862
00:32:57,993 --> 00:32:59,962
MODELS AND YOU CAN GET

863
00:32:59,962 --> 00:33:01,597
INGRAFTMENT AND YOU CAN KEEP

864
00:33:01,597 --> 00:33:05,000
THIS OVERTIME BUT MDS, IF PEOPLE

865
00:33:05,000 --> 00:33:07,136
HAD 0.5% INGRAFTMENT, THEY WERE

866
00:33:07,136 --> 00:33:08,804
HAPPY THEY HAD INGRAFTMENT AND

867
00:33:08,804 --> 00:33:12,274
YOU COULD STUDY IT A LITTLE BIT.

868
00:33:12,274 --> 00:33:14,910
SO WHAT RICHARD'S GROUP DID, WAS

869
00:33:14,910 --> 00:33:17,613
TOO LOOK AT, YOU KNOW CYTOKINES

870
00:33:17,613 --> 00:33:19,515
AND HEMEAT O POETICESIS AND WHAT

871
00:33:19,515 --> 00:33:21,350
CYTOKINES ARE NEEDED AND TO LOOK

872
00:33:21,350 --> 00:33:26,889
AT WELL, WHERE IS A CROSS

873
00:33:26,889 --> 00:33:28,023
REACTIVITY BETWEEN MOUSE AND

874
00:33:28,023 --> 00:33:29,125
HUMAN, RIGHT IN AND MOST

875
00:33:29,125 --> 00:33:31,660
IMPORTANTLY THEY WERE LOOKING AT

876
00:33:31,660 --> 00:33:34,029
OKAY, WHICH CYTOKINES FROM THE

877
00:33:34,029 --> 00:33:44,473
MOUSE DO NOT WORK ON HUMAN

878
00:33:45,508 --> 00:33:50,246
CELLS? 

879
00:33:50,246 --> 00:33:53,816
AND AMONG THOSE THROMPOETIN, AND

880
00:33:53,816 --> 00:34:00,990
OTHERS, AND THEY PAIRED UP WITH

881
00:34:00,990 --> 00:34:02,491
REGEN RON WHO DEVELOPED THESE

882
00:34:02,491 --> 00:34:05,294
TECHNOLOGIES SO IT WAS LIKE JUST

883
00:34:05,294 --> 00:34:06,228
BEFORE CRISPR SOLVED EVERYTHING

884
00:34:06,228 --> 00:34:08,964
AND THEY INTRODUCE THE ENTIRE

885
00:34:08,964 --> 00:34:12,001
HUMAN GENOMIC LOCUST INTO THE

886
00:34:12,001 --> 00:34:13,035
MOUSE GENOMIC LOCUST.

887
00:34:13,035 --> 00:34:16,305
SO NOW THE ENTIRE CODING AND

888
00:34:16,305 --> 00:34:20,042
ALSO NONCODING BUT EVER SEQUENCE

889
00:34:20,042 --> 00:34:22,711
OF THE HUMAN GENES WERE

890
00:34:22,711 --> 00:34:24,180
PHYSIOLOGICALLY EXPRESSED UNDER

891
00:34:24,180 --> 00:34:26,982
THE MOUSE, YOU KNOW REGULATORY

892
00:34:26,982 --> 00:34:27,383
SEQUENCES.

893
00:34:27,383 --> 00:34:28,751
AND THIS IS KIND OF JUST A

894
00:34:28,751 --> 00:34:32,288
LITTLE BIT OF A STORY OF THESE,

895
00:34:32,288 --> 00:34:33,789
YOU KNOW HUMANIZED--IMMUNO

896
00:34:33,789 --> 00:34:34,623
DEFICIENT MODELS, RIGHT, SO I

897
00:34:34,623 --> 00:34:40,062
REMEMBER WHEN I WAS IN--AT CHLA,

898
00:34:40,062 --> 00:34:43,399
YOU KNOW PEOPLE WERE WORKING ON

899
00:34:43,399 --> 00:34:45,367
NUDE AND SKID MICE, SO WE FIRST

900
00:34:45,367 --> 00:34:47,136
PRIORITY YOU HAVE TO HAVE A

901
00:34:47,136 --> 00:34:48,404
MOUSE THAT IS IMMUNO DEFICIENT

902
00:34:48,404 --> 00:34:50,406
SO THE MOUSE IMMUNE CELLS DO NOT

903
00:34:50,406 --> 00:34:53,509
ATTACK THE HUMAN CELLS AND YOU

904
00:34:53,509 --> 00:34:56,946
KNOW STARTED WITH GETTING RID OF

905
00:34:56,946 --> 00:34:59,181
T-CELLS, T AND B CELLS, MAYBE

906
00:34:59,181 --> 00:35:04,653
INHIBITING A LITTLE BIT

907
00:35:04,653 --> 00:35:05,955
MONOCYTES MACROPHAGES AND THEN

908
00:35:05,955 --> 00:35:07,690
EVENTUALLY ALSO GETTING RID OF

909
00:35:07,690 --> 00:35:09,458
NK CELLS, SO NOW THE HUMAN CELLS

910
00:35:09,458 --> 00:35:11,460
CAN GET IN BUT AGAIN, RIGHT, SO

911
00:35:11,460 --> 00:35:13,762
THEY WERE STILL LACKING

912
00:35:13,762 --> 00:35:14,363
DIFFERENTIATION FACTORS THAT,

913
00:35:14,363 --> 00:35:16,799
WOOED ON THEM, SO THEN A VERY

914
00:35:16,799 --> 00:35:20,135
POPULAR MODEL IS NSGS MODEL AND

915
00:35:20,135 --> 00:35:22,905
THAT EXPRESSES STEM CELL

916
00:35:22,905 --> 00:35:24,506
FACTORED GMCSF AND INTERLEUKIN

917
00:35:24,506 --> 00:35:26,709
3, BUT THOSE ARE TRANSGENIC MICE

918
00:35:26,709 --> 00:35:29,345
SO THOSE FACTORS ARE JUST

919
00:35:29,345 --> 00:35:31,313
EXPRESSED ANYWHERE, THEY'RE NOT

920
00:35:31,313 --> 00:35:32,248
REGULATED, THEY'RE EXPRESSED ALL

921
00:35:32,248 --> 00:35:35,184
THE TIME SO ACTUALLY IF YOU

922
00:35:35,184 --> 00:35:36,018
INGRAFT HEMATOPOIETIC STEM CELLS

923
00:35:36,018 --> 00:35:38,754
INTO THE MICE FOR SOMETIME, YOU

924
00:35:38,754 --> 00:35:41,624
GET GET HEMEAT O POEESIS BUT

925
00:35:41,624 --> 00:35:43,492
THEN YOU GET EXHAUSTION BECAUSE

926
00:35:43,492 --> 00:35:44,693
OF THESE CELL LINES.

927
00:35:44,693 --> 00:35:46,962
AND THEN THE LAB LABELED THESE

928
00:35:46,962 --> 00:35:49,164
MICE MR. G, IT'S A GOOD ACRONYM

929
00:35:49,164 --> 00:35:50,399
AND THESE THAT STAND FOR THE

930
00:35:50,399 --> 00:35:53,102
CYTOKINES THAT ARE INTRODUCED,

931
00:35:53,102 --> 00:35:55,137
THE RAD GAMMA KNOCK OUT

932
00:35:55,137 --> 00:35:57,740
PHENOTYPE, IMMUNO PHENOTYPE AND

933
00:35:57,740 --> 00:36:00,175
THERE'S ALSO REPLACEMENT OF

934
00:36:00,175 --> 00:36:04,280
ALPHA WITH HUMAN ALPHA, AND

935
00:36:04,280 --> 00:36:06,015
ALPHA BINDS CD47, IT'S KIND OF

936
00:36:06,015 --> 00:36:07,616
THIS DON'T EAT ME SIGNAL AND

937
00:36:07,616 --> 00:36:15,658
IT'S VERY IMPORTANT IN THE  XENO

938
00:36:15,658 --> 00:36:15,925
TOLERANCE.

939
00:36:15,925 --> 00:36:18,560
SO MANY POST DOCS WORKING ON

940
00:36:18,560 --> 00:36:21,096
THIS IN RICHARD'S LAB, REALLY

941
00:36:21,096 --> 00:36:23,632
CHARACTERIZED EVERY SINGLE

942
00:36:23,632 --> 00:36:25,834
REPLACEMENT, AGAIN, RESULTING IN

943
00:36:25,834 --> 00:36:27,102
THE CYTOKINE KNOCK OUT MOUSE

944
00:36:27,102 --> 00:36:28,871
MODEL THAT ALSO INCLUDES

945
00:36:28,871 --> 00:36:30,572
INTERLEUKIN 6, THERE ARE OTHER

946
00:36:30,572 --> 00:36:32,174
VERSIONS FOR EXAMPLE,

947
00:36:32,174 --> 00:36:33,575
INTERLEUKIN 15 THAT THEY QUICKLY

948
00:36:33,575 --> 00:36:35,611
REALIZED THAT IF YOU HAVE MCSF

949
00:36:35,611 --> 00:36:37,379
AND YOU HAVE MONOCYTES AND

950
00:36:37,379 --> 00:36:40,349
MACROPHAGES, THOSE ACTUALLY THEN

951
00:36:40,349 --> 00:36:42,017
MAKE HUMAN IL15 THAT'S NEEDED

952
00:36:42,017 --> 00:36:43,285
FOR NK CELL DEVELOPMENT, YOU

953
00:36:43,285 --> 00:36:46,422
DON'T NEED TO PUT IN EXOGENOUS

954
00:36:46,422 --> 00:36:47,356
INTERLEUKIN 15 AND SO THEN,

955
00:36:47,356 --> 00:36:49,425
EARLY ON I WAS STILL A POST DOC

956
00:36:49,425 --> 00:36:52,261
BUT IT WAS EVENTUALLY FINISHED

957
00:36:52,261 --> 00:36:54,096
BY WOREALLY MAIRKSZING PEOPLE IN

958
00:36:54,096 --> 00:36:56,465
MY LAB, BILLY AND

959
00:36:56,465 --> 00:36:57,566
[INDISCERNIBLE] AND TOGETHER

960
00:36:57,566 --> 00:36:59,802
WITH ANTHONY [INDISCERNIBLE], WE

961
00:36:59,802 --> 00:37:01,570
TOOK MDS OF ALL LO GRADE AND

962
00:37:01,570 --> 00:37:05,674
HIGH RISK AND THEN ALWAYS

963
00:37:05,674 --> 00:37:07,176
COMPARED INGRAFTMENT AND MR. G

964
00:37:07,176 --> 00:37:09,411
MICE COMPARED TO THE GOAL

965
00:37:09,411 --> 00:37:11,613
STANDARD THEIR, MSG MICE AND HAD

966
00:37:11,613 --> 00:37:13,682
MUCH, MUCH, IMPROVED INGRAFTMENT

967
00:37:13,682 --> 00:37:15,351
AND REALLY OVER50% OF THE MICE

968
00:37:15,351 --> 00:37:16,418
INGRAFTMENT OVER LIKE 10%, SO

969
00:37:16,418 --> 00:37:18,754
YOU CAN IMAGINE, YOU CAN REALLY

970
00:37:18,754 --> 00:37:20,155
ALSO TEST DRUGS BECAUSE CAN YOU

971
00:37:20,155 --> 00:37:22,157
ACTUALLY MEASURE SOMETHING WITH

972
00:37:22,157 --> 00:37:24,426
SOME RELIABILITY.

973
00:37:24,426 --> 00:37:26,595
IMPORTANTLY, WE COULD RECEDE

974
00:37:26,595 --> 00:37:28,097
DISPLASSIA, BECAUSE YOU KNOW YOU

975
00:37:28,097 --> 00:37:30,766
GET ARITHROUGH ATOM POETICESIS,

976
00:37:30,766 --> 00:37:32,968
THE HALLMARK, WITH THE

977
00:37:32,968 --> 00:37:34,837
MUTATIONS, AND I REMEMBER, YOU

978
00:37:34,837 --> 00:37:35,804
KNOW   PERFORMING A BONE MARROW

979
00:37:35,804 --> 00:37:37,806
ON THE PATIENT, SEEING THE BONE

980
00:37:37,806 --> 00:37:39,308
MARROW IN PATHOLOGY, AND THEN

981
00:37:39,308 --> 00:37:40,976
THE PEASHT GRACIOUSLY WRITE,

982
00:37:40,976 --> 00:37:42,845
CONSENTED TO GIVING US A SAMPLE,

983
00:37:42,845 --> 00:37:43,812
INGRAFTING IT INTO MICE AND

984
00:37:43,812 --> 00:37:45,447
LOOKING AT THE MOUSE, AND I'M

985
00:37:45,447 --> 00:37:48,984
LIKE WOW, THIS IS EXACTLY THE

986
00:37:48,984 --> 00:37:50,386
SAME PHENOTYPE.

987
00:37:50,386 --> 00:37:51,487
WE ALSO SEQUENCED, THE PATIENTS

988
00:37:51,487 --> 00:37:54,923
AND THE MICE TO SHOW THAT WE ARE

989
00:37:54,923 --> 00:37:57,192
INGRAFTING THE MUTANT STEM

990
00:37:57,192 --> 00:37:59,661
CELLS, AND THEN ULTIMATELY WE

991
00:37:59,661 --> 00:38:00,929
ALSO   PERFORMED THERAPEUTIC

992
00:38:00,929 --> 00:38:03,198
TESTING WITH MUTANT IDH

993
00:38:03,198 --> 00:38:05,467
INHIBITORS COMPARING THAT FOR

994
00:38:05,467 --> 00:38:08,570
EXAMPLE IN ELABORATE OR PERP

995
00:38:08,570 --> 00:38:11,407
INHIBITOR WHICH A COLLEAGUE HAD

996
00:38:11,407 --> 00:38:13,609
SHOWN WORKS IN IDH MUTANT CELLS

997
00:38:13,609 --> 00:38:18,881
THAT END UP HAVING A DNA REPAIR

998
00:38:18,881 --> 00:38:19,114
DEFECT.

999
00:38:19,114 --> 00:38:20,616
SO WE'RE VERY EXCITED ABOUT THAT

1000
00:38:20,616 --> 00:38:22,618
AND THEN OF COURSE, ONCE YOU

1001
00:38:22,618 --> 00:38:24,153
HAVE SUCH A MODEL THAT'S QUITE

1002
00:38:24,153 --> 00:38:25,854
GOOD, YOU THINK IT'S NOT GOOD

1003
00:38:25,854 --> 00:38:26,121
ENOUGH.

1004
00:38:26,121 --> 00:38:29,892
AND 1 BIG DEFICIT IN ALL

1005
00:38:29,892 --> 00:38:31,527
HUMANIZED MOUSE MODELS IS THAT

1006
00:38:31,527 --> 00:38:34,663
WHILE YOU STILL HAVE THE MOUSE

1007
00:38:34,663 --> 00:38:36,598
INNATE IMMUNITY, SO THE MOUSE

1008
00:38:36,598 --> 00:38:38,167
GRANUE LOW SIGHTS BUT THE

1009
00:38:38,167 --> 00:38:39,001
MACROPHAGES ARE ACTUALLY A

1010
00:38:39,001 --> 00:38:41,737
PROBLEM, AND SO IN THESE MICE

1011
00:38:41,737 --> 00:38:44,139
YOU ACTUALLY DON'T HAVE

1012
00:38:44,139 --> 00:38:45,140
CIRCULATING RED CELLS, AND

1013
00:38:45,140 --> 00:38:49,411
INKRED EBLLY ARK MAZING POST DOC

1014
00:38:49,411 --> 00:38:53,348
IN RICHARD'S LAB WHOSE GOT

1015
00:38:53,348 --> 00:38:56,018
FACULTY 

1016
00:38:56,018 --> 00:38:58,320
FACULTY TRY TO FIGURE OUT AGAIN

1017
00:38:58,320 --> 00:38:59,621
DELETING ALL THINGS FROM THE

1018
00:38:59,621 --> 00:39:02,391
MICE, CROSSING IT TO MCSF,

1019
00:39:02,391 --> 00:39:03,225
KNOCKOUT MICE, ET CETERA TO SEE

1020
00:39:03,225 --> 00:39:06,128
WHAT WE CAN GET, CAN WE GET

1021
00:39:06,128 --> 00:39:09,631
CIRCULATING HUMAN RED CELLS, AND

1022
00:39:09,631 --> 00:39:10,899
THEN 1 EXPERIMENT THAT KIND OF

1023
00:39:10,899 --> 00:39:12,868
TOLD THEM WHAT TO GO AFTER WAS

1024
00:39:12,868 --> 00:39:15,104
TO TAKE HUMAN RED CELLS, LABEL

1025
00:39:15,104 --> 00:39:16,171
THEM WITH FLUORESCENT DIE,

1026
00:39:16,171 --> 00:39:17,806
INYECT THEM INTO THE MICE AND

1027
00:39:17,806 --> 00:39:19,208
TAKING OTHER ORGANS AND THEN

1028
00:39:19,208 --> 00:39:21,076
LOOKING WHERE ARE THESE CELLS,

1029
00:39:21,076 --> 00:39:22,444
SO SOME GOT THE QUESTION THE

1030
00:39:22,444 --> 00:39:24,646
SPLEEN AND WE ALL THOUGHT THAT

1031
00:39:24,646 --> 00:39:26,748
WAS THE MAIN SITE WHERE MOST OF

1032
00:39:26,748 --> 00:39:28,183
THE CELLS WERE SEQUESTERED IN

1033
00:39:28,183 --> 00:39:32,321
THE LIVER  LIVER EMPLOY SO THERS

1034
00:39:32,321 --> 00:39:32,821
THIS MOUSE.

1035
00:39:32,821 --> 00:39:34,223
I ALWAYS HAVE TO WRITE THIS DOWN

1036
00:39:34,223 --> 00:39:37,593
BECAUSE I CAN NEVER SAY IT THAT

1037
00:39:37,593 --> 00:39:45,033
IS KNOCKED OUT FOR FUMER OLDER

1038
00:39:45,033 --> 00:39:46,101
PEOPLE ASEITATE-ASIGNIFYTAL

1039
00:39:46,101 --> 00:39:46,635
HYDROLACE [INDISCERNIBLE].

1040
00:39:46,635 --> 00:39:51,607
SO THEY HAVE A METABOLIC DEFECT

1041
00:39:51,607 --> 00:39:52,674
IN TYROSEEN METABOLISM AND SO

1042
00:39:52,674 --> 00:39:53,942
YOU ACTUALLY HAVE TO KEEP THEM

1043
00:39:53,942 --> 00:39:58,380
ON SPECIAL WATER SO THEY DON'T

1044
00:39:58,380 --> 00:39:59,281
GENERATE TOXIC METABOLITES THAT

1045
00:39:59,281 --> 00:40:00,249
TISSUE CULTURE MEDIAS THEIR

1046
00:40:00,249 --> 00:40:02,417
LIVER CELLS.

1047
00:40:02,417 --> 00:40:05,521
WELL, IT TURNS OUT YOU CAN

1048
00:40:05,521 --> 00:40:07,256
ACTUALLY TAKE HEPATOCYTES,

1049
00:40:07,256 --> 00:40:09,324
EITHER MOUSE HEPATOCYTES OR

1050
00:40:09,324 --> 00:40:12,394
HUMAN HEPATOCYTES AND INJECT

1051
00:40:12,394 --> 00:40:15,030
THEM INTO THE SPLENIC VEIN, AND

1052
00:40:15,030 --> 00:40:17,566
THEN THESE CELLS WERE LARGE IN

1053
00:40:17,566 --> 00:40:20,903
THE LIVER, SO YOU CAN INJECT

1054
00:40:20,903 --> 00:40:21,637
HUMAN HEPATOCYTES INTO THESE

1055
00:40:21,637 --> 00:40:24,039
SURGERY INTO THESE MICE, AND YOU

1056
00:40:24,039 --> 00:40:25,674
WITHDRAW THIS WATER, SO WITH

1057
00:40:25,674 --> 00:40:28,911
BITS NOW, YOU'RE CREATING THE

1058
00:40:28,911 --> 00:40:30,179
MOUSE HUMAN HEPATOCYTES AND THEY

1059
00:40:30,179 --> 00:40:32,247
WILL ACTUALLY TAKE OVER AND THEN

1060
00:40:32,247 --> 00:40:34,483
YOU CAN MEASURE HUMAN ALBUT MIN

1061
00:40:34,483 --> 00:40:36,018
IN PLASMA AND KIND OF WHAT IT

1062
00:40:36,018 --> 00:40:39,888
SETS A CERTAIN LEVEL, THEY'RE

1063
00:40:39,888 --> 00:40:41,256
PROBABLY MOSTLY HUMAN

1064
00:40:41,256 --> 00:40:43,692
HEPATOCYTES AROUND AND SO THEN

1065
00:40:43,692 --> 00:40:48,797
WE CAN TAKE THESE HUMAN LIVER

1066
00:40:48,797 --> 00:40:52,000
HUMANIZED MR. G MICE AND INGRAFT

1067
00:40:52,000 --> 00:40:56,004
HUMAN CD34 CELLS JUST

1068
00:40:56,004 --> 00:40:57,506
RETROORBITALLY, AND NOW WE HAVE

1069
00:40:57,506 --> 00:41:00,976
HUMAN AND MICE THAT HAVE HUMAN

1070
00:41:00,976 --> 00:41:04,713
LIVER AND HEMATOPOIETIC STEM

1071
00:41:04,713 --> 00:41:06,148
CELLS INGRAFTED AND REALLY TO

1072
00:41:06,148 --> 00:41:08,016
OUR DELIGHT, THESE MICE NOW HAVE

1073
00:41:08,016 --> 00:41:09,084
CIRCULATING HUMAN RED CELLS.

1074
00:41:09,084 --> 00:41:10,953
AGAIN THIS IS A LOG SCALE, BUT

1075
00:41:10,953 --> 00:41:13,755
YOU KNOW REALLY THERE ARE UP TO

1076
00:41:13,755 --> 00:41:16,291
10% CIRCULATING HUMAN RED CELLS,

1077
00:41:16,291 --> 00:41:16,959
IN THESE MICE.

1078
00:41:16,959 --> 00:41:18,961
YOU CAN'T REPLACE ALL THE RED

1079
00:41:18,961 --> 00:41:20,395
CELLS, THE HUMAN RED CELLS ARE

1080
00:41:20,395 --> 00:41:21,797
MUCH BIGGER THAN THE RED CELLS

1081
00:41:21,797 --> 00:41:24,199
AND THE MICE NEED THEIR OWN RED

1082
00:41:24,199 --> 00:41:27,236
CELLS THAT CIRCULATE PROPERLY,

1083
00:41:27,236 --> 00:41:28,370
MOST LIKELY.

1084
00:41:28,370 --> 00:41:29,571
AND THAT WAS--WELL, SURPRISINGLY

1085
00:41:29,571 --> 00:41:31,773
ALSO THAT WE ACTUALLY HAD HIGHY

1086
00:41:31,773 --> 00:41:34,476
INGRAFTMENT IN THE BONE MARROW

1087
00:41:34,476 --> 00:41:36,445
AND ALSO GENERALLY MORE ERYTH

1088
00:41:36,445 --> 00:41:38,013
ROUGH ATOM PORESIS IN THE BONE

1089
00:41:38,013 --> 00:41:39,081
MARROW, SO SUGGESTING IN THE

1090
00:41:39,081 --> 00:41:40,482
LIVER, MANY OTHER PROTEINS THAT

1091
00:41:40,482 --> 00:41:44,052
MAY BE 1 DAY WE COULD SCREEN, TO

1092
00:41:44,052 --> 00:41:45,587
UNDERSTAND, WHAT OTHER PROTEINS

1093
00:41:45,587 --> 00:41:48,690
MADE IN THE LIVER THAT ARE

1094
00:41:48,690 --> 00:41:51,226
ACTUALLY IMPORTANT FOR HEMEAT O

1095
00:41:51,226 --> 00:41:51,560
POETICESIS.

1096
00:41:51,560 --> 00:41:56,665
SO, WE WANTED TO KNOW, YOU KNOW

1097
00:41:56,665 --> 00:41:58,400
WHAT THE MECHANISM IS FOR THIS

1098
00:41:58,400 --> 00:41:59,835
IMPROVED, YOU KNOW CIRCULATION

1099
00:41:59,835 --> 00:42:02,804
OF HUMAN RED CELL, AND 1 VERY

1100
00:42:02,804 --> 00:42:04,406
OBVIOUS THING WAS ACTUALLY TO

1101
00:42:04,406 --> 00:42:07,009
LOOK FOR COMPLEMENT AND SO

1102
00:42:07,009 --> 00:42:11,446
REALLY THE LIVER HUMANIZED MICE

1103
00:42:11,446 --> 00:42:15,417
HAVE LESS MURINE C3 AND THERE'S

1104
00:42:15,417 --> 00:42:17,185
ALSO LESS COMPLEMENT COATING ON

1105
00:42:17,185 --> 00:42:18,687
THE HUMAN RED CELLS SO YOU

1106
00:42:18,687 --> 00:42:20,055
REALLY JUST DON'T HAVE

1107
00:42:20,055 --> 00:42:21,356
DESTRUCTION OF THE LABELING OF

1108
00:42:21,356 --> 00:42:23,759
THE RED CELLS WITH COMPLEMENT

1109
00:42:23,759 --> 00:42:29,564
AND THEN THEIR PHAGOCYTOSIS BY

1110
00:42:29,564 --> 00:42:31,767
MOUSE MACROPHAGES AND SO I THINK

1111
00:42:31,767 --> 00:42:34,970
THIS RIGHT--I KNOW HAD IS CANCER

1112
00:42:34,970 --> 00:42:38,273
BUT ACTUALLY ATL, BUT CLASSIC

1113
00:42:38,273 --> 00:42:39,308
[INDISCERNIBLE] AND ALSO OUR

1114
00:42:39,308 --> 00:42:40,275
PATIENTS WITH SICKLE CELL

1115
00:42:40,275 --> 00:42:43,211
DISEASE ARE GETTING CARE IN THE

1116
00:42:43,211 --> 00:42:44,746
CENTER, AND WHICH IS PHENOMENAL

1117
00:42:44,746 --> 00:42:46,515
AND THE RIGHT THING TO DID.

1118
00:42:46,515 --> 00:42:49,051
SO WE TOOK THESE CELLS AND CD34

1119
00:42:49,051 --> 00:42:51,720
CELLS WITH PATIENTS FROM SICKLE

1120
00:42:51,720 --> 00:42:53,722
CELL DISEASE, A MONOGENIC DEC WE

1121
00:42:53,722 --> 00:42:54,790
IGNORED FOR FAR TOO LONG AND

1122
00:42:54,790 --> 00:42:57,626
THEN WHAT YOU SEE NOW HERE, ARE

1123
00:42:57,626 --> 00:43:01,263
ACTUALLY SECTIONS FROM LUNG,

1124
00:43:01,263 --> 00:43:05,734
LIVER, AND KIDNEY WITH THE HUMAN

1125
00:43:05,734 --> 00:43:06,635
SPECIFIC HEMOGLOBIN A ANTIBODY

1126
00:43:06,635 --> 00:43:09,204
AND IF YOU COMPARE LUNGS FROM

1127
00:43:09,204 --> 00:43:11,506
PATIENTS FROM MICE INGRAFTED

1128
00:43:11,506 --> 00:43:15,310
WITH THESE CELLS, OR CD34 CELLS

1129
00:43:15,310 --> 00:43:17,012
WITH PATIENT FROM SICKLE CELL

1130
00:43:17,012 --> 00:43:19,047
DISEASE, CAN YOU SEE VASE O,

1131
00:43:19,047 --> 00:43:21,049
CLIEWGZ, SO THESE ARE RED CELLS

1132
00:43:21,049 --> 00:43:24,553
STUCK IN THE LUNG VASCULATURE,

1133
00:43:24,553 --> 00:43:27,789
YOU CAN SEE RED CELLS KIND OF

1134
00:43:27,789 --> 00:43:28,857
STUCK IN THE CAPILLARIES IN THE

1135
00:43:28,857 --> 00:43:31,493
LIVER AND ALSO IN THE GLUE MARUE

1136
00:43:31,493 --> 00:43:33,495
LIE AND YOU KNOW IN THE KIDNEYS

1137
00:43:33,495 --> 00:43:37,566
SO WE CAN ACTUALLY NOW STUDY,

1138
00:43:37,566 --> 00:43:43,605
THESE OCCLUSION FROM HUMAN

1139
00:43:43,605 --> 00:43:43,939
CELLS.

1140
00:43:43,939 --> 00:43:47,142
SO AGAIN WE'RE NEVER SATISFIED.

1141
00:43:47,142 --> 00:43:48,744
SO THAT IS ALSO COMPLICATED

1142
00:43:48,744 --> 00:43:52,547
MODEL TO GENERATE BUT YOU KNOW 1

1143
00:43:52,547 --> 00:43:54,649
THEORY IS THAT IN MDS,

1144
00:43:54,649 --> 00:43:55,951
INFLAMMATION PLAYS A ROLE,

1145
00:43:55,951 --> 00:43:56,651
RIGHT?

1146
00:43:56,651 --> 00:43:58,887
AND WE KNOW INFLAMMATION PLAYS A

1147
00:43:58,887 --> 00:44:02,758
ROLE AND ET CETERA, AND SO TO

1148
00:44:02,758 --> 00:44:04,126
INGRAFT HUMAN WITH THESE MICE WE

1149
00:44:04,126 --> 00:44:07,195
HAVE TO RADIATE THE MICE.

1150
00:44:07,195 --> 00:44:13,268
THERE ARE BEAUTIFUL PUBLICATIONS

1151
00:44:13,268 --> 00:44:20,342
SHOWINGS THAT IF YOU MUTATE THE

1152
00:44:20,342 --> 00:44:21,443
STEM CELL FACTOR RECEPTOR IN

1153
00:44:21,443 --> 00:44:23,412
MICE THAT YOU MAY NOT NEED TO

1154
00:44:23,412 --> 00:44:28,016
RADIATE BUT THAT IS ENOUGH OF A

1155
00:44:28,016 --> 00:44:30,051
COMPETITIVE DISADVANTAGE TO THE

1156
00:44:30,051 --> 00:44:32,287
MURINE STEM CELLS AND PORESIS

1157
00:44:32,287 --> 00:44:37,692
THAT YOU CAN NOW INGRAFT HUMAN

1158
00:44:37,692 --> 00:44:40,262
CELLS WITHOUT IRRADIATION, SO

1159
00:44:40,262 --> 00:44:42,431
VIA CRISPR, AGAIN, WE GENERATED

1160
00:44:42,431 --> 00:44:43,965
THESE MICE WITH THESE W41

1161
00:44:43,965 --> 00:44:45,500
MUTATIONS SO THESE ARE ACTUALLY

1162
00:44:45,500 --> 00:44:47,569
MUTATIONS THAT WERE DRIEBED IN

1163
00:44:47,569 --> 00:44:49,304
B6 MICE CALLED THE WHITE SPOTTED

1164
00:44:49,304 --> 00:44:51,106
MICE SO IN EFFECT THE RECEPTOR

1165
00:44:51,106 --> 00:44:52,707
IS ALSO IMPORTANT TO THE NANO

1166
00:44:52,707 --> 00:44:54,109
SIGHTS SO THESE MICE ARE

1167
00:44:54,109 --> 00:44:55,710
ACTUALLY BLACK BUT THEY HAVE A

1168
00:44:55,710 --> 00:44:57,112
LITTLE WHITE SPOT ON THEIR

1169
00:44:57,112 --> 00:45:00,115
STOMACH, THAT WAS EASY FOR GENO

1170
00:45:00,115 --> 00:45:03,819
TYPING, THE MSG IS WHITE SO WE

1171
00:45:03,819 --> 00:45:04,753
SELECTED COMPLICATED GEANY

1172
00:45:04,753 --> 00:45:07,055
TYPING TO MAKE THESE MICE AND SO

1173
00:45:07,055 --> 00:45:11,359
WITH THIS MOUSE MODEL WE STARTED

1174
00:45:11,359 --> 00:45:15,997
A COLLABORATION WITH LEE GRIMES

1175
00:45:15,997 --> 00:45:18,467
AND [INDISCERNIBLE], AND HE'S A

1176
00:45:18,467 --> 00:45:21,102
PHENOMENAL INFORMATICS PERSON TO

1177
00:45:21,102 --> 00:45:21,937
MODEL MYELODYSPLASIA IN THESE

1178
00:45:21,937 --> 00:45:24,372
MICE AND IT REALLY WAS THE IDEA,

1179
00:45:24,372 --> 00:45:26,208
WITH HOW FAITHFUL REALLY ARE

1180
00:45:26,208 --> 00:45:30,612
THESE MODELS, WE HAD DONE LIKE A

1181
00:45:30,612 --> 00:45:33,949
CRUDE ASSESSMENT AND WITH LEE,

1182
00:45:33,949 --> 00:45:41,923
AMY AND GABRIEL AMAZING PEOPLE,

1183
00:45:41,923 --> 00:45:43,992
SEAN, JENNIFER AND WEI, SO LEE

1184
00:45:43,992 --> 00:45:47,429
IN THE PAPER, HIS PAPER ON THIS

1185
00:45:47,429 --> 00:45:49,598
PHENOMENON METHOD JUST PUBLISHED

1186
00:45:49,598 --> 00:45:52,701
IN IMMUNOLOGY, HE DEVELOPED A

1187
00:45:52,701 --> 00:45:54,503
VERY COMPREHENSIVE SIDE SEQ

1188
00:45:54,503 --> 00:45:56,505
PANEL FOR HEMEAT O POETICESIS SO

1189
00:45:56,505 --> 00:45:59,174
EVERY ANTIBODY IS TITRATED.

1190
00:45:59,174 --> 00:46:01,910
AND SO, AND YOU SEE, HIS GROUP

1191
00:46:01,910 --> 00:46:05,614
SEQUENCED HEPATITIS E C CONTROL

1192
00:46:05,614 --> 00:46:07,916
[INDISCERNIBLE], CD34 CELLS, MSC

1193
00:46:07,916 --> 00:46:09,451
CELLS AND, IT NC AND THIS IS THE

1194
00:46:09,451 --> 00:46:10,418
WHOLE WITH BEAUTIFUL

1195
00:46:10,418 --> 00:46:10,785
REPRESENTATION.

1196
00:46:10,785 --> 00:46:14,756
THIS IS NOW AN MDS SAMPLE FROM

1197
00:46:14,756 --> 00:46:17,392
JOHNS HOPKINS AND WE TOOK THE

1198
00:46:17,392 --> 00:46:21,396
SASM AND THE G-MICE, TO AFTER 16

1199
00:46:21,396 --> 00:46:23,265
WEEKS AFTER INGRAFTMENT,

1200
00:46:23,265 --> 00:46:24,466
HARVESTED THE BONE MARROW TO

1201
00:46:24,466 --> 00:46:27,068
SEQUENCE AND YOU CAN SEE THAT

1202
00:46:27,068 --> 00:46:28,937
WHY THIS IS NOT PERFECT, IT'S

1203
00:46:28,937 --> 00:46:31,273
QUITE A GOOD REPRESENTATION, AT

1204
00:46:31,273 --> 00:46:34,109
THE SINGLE CELL LEVEL OF THE MDS

1205
00:46:34,109 --> 00:46:34,910
POPULATIONS AND HERE'S PROBABLY

1206
00:46:34,910 --> 00:46:38,346
A LITTLE BIT CLEARER SO THIS IS

1207
00:46:38,346 --> 00:46:39,981
THEN SEQUENCING RIGHTFULLY SO

1208
00:46:39,981 --> 00:46:41,383
ALL YEENS, THESE ARE ALL

1209
00:46:41,383 --> 00:46:44,686
SUBPOPULATIONS AND YOU CAN SEE

1210
00:46:44,686 --> 00:46:46,488
THAT THE MDS XENOGRAFTS LOOK

1211
00:46:46,488 --> 00:46:48,990
LIKE THE PASHT IN TERLS OF

1212
00:46:48,990 --> 00:46:51,993
DIFFERENT YALE EXPRESSION

1213
00:46:51,993 --> 00:46:52,794
ANALYSIS.

1214
00:46:52,794 --> 00:46:54,663
SO NOW WE HAVE SO 50ICATEED

1215
00:46:54,663 --> 00:46:58,233
MODEL WHERE WE CAN INGRAFT, AND

1216
00:46:58,233 --> 00:47:00,936
1 OF THE GOALS NOW IS TO INGRAFT

1217
00:47:00,936 --> 00:47:06,608
MICE AND TREAT THEM WITHAISE O

1218
00:47:06,608 --> 00:47:08,243
CYTODEAN, THE STANDARD OF CLAIR

1219
00:47:08,243 --> 00:47:11,880
THAT I THINK ANY MDS CLINICIAN

1220
00:47:11,880 --> 00:47:13,715
DOESN'T LIKE AT ALL BECAUSE WE

1221
00:47:13,715 --> 00:47:16,017
DON'T KNOW WHAT IT DOES OR HOW

1222
00:47:16,017 --> 00:47:18,219
IT WORKS AND MANY PATIENTS

1223
00:47:18,219 --> 00:47:19,621
RELAPSE EVEN AN TRANSPLANT AND

1224
00:47:19,621 --> 00:47:25,860
ONCE PATIENTS ARE REFRACTORY TO

1225
00:47:25,860 --> 00:47:26,828
ISOTOPE AZACYTIDINE, AND THERE'S

1226
00:47:26,828 --> 00:47:30,932
NOTHING ELSE THAT WORKS.

1227
00:47:30,932 --> 00:47:34,536
SO MICHELLE RUDEK YOU KNOW

1228
00:47:34,536 --> 00:47:36,171
DEVELOPED THE REGIMEN IN THE

1229
00:47:36,171 --> 00:47:37,339
MR. G MICE, THEY ARE SENSITIVE,

1230
00:47:37,339 --> 00:47:40,976
AND WE WERE ABLE TO YOU KNOW

1231
00:47:40,976 --> 00:47:48,049
MEASURE BOTH INCORPORATION OF

1232
00:47:48,049 --> 00:47:50,819
THE NUTRITIGHT INTO DNA AND THEN

1233
00:47:50,819 --> 00:47:53,822
ALSO SHOW DECREASE IN

1234
00:47:53,822 --> 00:47:54,823
METHYLATION RIGHT HERE.

1235
00:47:54,823 --> 00:47:56,257
SO THAT'S AN ONGOING PROJECT.

1236
00:47:56,257 --> 00:47:58,126
I THINK I BRIEFLY SHOWED THIS,

1237
00:47:58,126 --> 00:48:00,629
WE HAD USED THESE MICE BEFORE

1238
00:48:00,629 --> 00:48:03,965
BECAUSE YOU KNOW WE HAD FOUND

1239
00:48:03,965 --> 00:48:07,836
THIS MECHANISM, THAT IDH MUTANT

1240
00:48:07,836 --> 00:48:09,437
MDS, AML IS ACTUALLY SENSITIVE

1241
00:48:09,437 --> 00:48:12,374
TO P A RP INHIBITION BECAUSE OF

1242
00:48:12,374 --> 00:48:14,909
A DNA REDETECTED AND THESE MICE

1243
00:48:14,909 --> 00:48:22,951
WERE INGRAFTED AND MDS AML AND

1244
00:48:22,951 --> 00:48:25,053
THEN THEY ARE--WE ACTUALLY  

1245
00:48:25,053 --> 00:48:25,654
PERFORMED BONE MARROW

1246
00:48:25,654 --> 00:48:26,921
ASSPIRITUAL STRUGGLESSATIONS

1247
00:48:26,921 --> 00:48:28,223
BEFORE TREATMENT AND SACRIFICE

1248
00:48:28,223 --> 00:48:30,425
MICE AFTER TREATMENT AND THIS IS

1249
00:48:30,425 --> 00:48:33,128
SHOWING IN THESE 2 PATIENTS,

1250
00:48:33,128 --> 00:48:37,198
RIGHT, IN THE SPECIFIC MUTANT

1251
00:48:37,198 --> 00:48:41,836
IDH INHIBITOR, OR THE LAB WORKS

1252
00:48:41,836 --> 00:48:43,471
REALLY WELL, AND THEN THIS IS A

1253
00:48:43,471 --> 00:48:47,776
PATIENT NOW WHO HAD SECONDARY

1254
00:48:47,776 --> 00:48:52,047
AML, AND WHEN ON TREATMENT WITH

1255
00:48:52,047 --> 00:48:53,548
AZATINIB FOR A WREER AND THEP

1256
00:48:53,548 --> 00:48:54,749
PROGRESSED TO AAMERICA L, AND

1257
00:48:54,749 --> 00:49:00,989
THEN WAS NO LONGER SENSITIVE TO

1258
00:49:00,989 --> 00:49:03,091
AZACITINETWORK IB, BUT WAS

1259
00:49:03,091 --> 00:49:03,892
SENSITIVE TO [INDISCERNIBLE].

1260
00:49:03,892 --> 00:49:06,294
AND THEN WENT TO TO A SMALL

1261
00:49:06,294 --> 00:49:07,495
TRIAL, AND THIS IS A WAKE UP

1262
00:49:07,495 --> 00:49:09,130
CALL THAT WE CAN DO A LOT OF

1263
00:49:09,130 --> 00:49:10,131
THINGS IN THESE MODELS IN

1264
00:49:10,131 --> 00:49:12,233
INVITRO IN THE PATIENT, GIVING

1265
00:49:12,233 --> 00:49:13,401
PATIENTS PREP WHEN THEY HAVE

1266
00:49:13,401 --> 00:49:15,470
AMERICA DS, IT LOOKED VERY

1267
00:49:15,470 --> 00:49:19,340
DIFFERENT AND I WOULD SAY, IT

1268
00:49:19,340 --> 00:49:20,175
PROBABLY DOESN'T WORK AND WE

1269
00:49:20,175 --> 00:49:22,477
HAVE A LOT MORE WORK TO DO TO

1270
00:49:22,477 --> 00:49:27,882
FIGURE OUT CAN 1 FOR EXAMPLE

1271
00:49:27,882 --> 00:49:28,850
INTRODUCE THESE TREATMENTS FOR

1272
00:49:28,850 --> 00:49:32,954
THESE PATIENTS OR MAYBE NOT,

1273
00:49:32,954 --> 00:49:34,222
BECAUSE MDS, CYTOPENIC IS NOT A

1274
00:49:34,222 --> 00:49:35,423
GOOD RUG TO USE.

1275
00:49:35,423 --> 00:49:37,292
BUT WE'RE REALLY POISED TO USE

1276
00:49:37,292 --> 00:49:39,527
THESE MODELS, USE OUR ADVANCES

1277
00:49:39,527 --> 00:49:40,729
IN UNDERSTANDING MUTATIONS TO

1278
00:49:40,729 --> 00:49:41,663
HOPEFULLY GET SOME TREATMENT TO

1279
00:49:41,663 --> 00:49:43,932
PATIENTS AND EVEN IF WE ONLY,

1280
00:49:43,932 --> 00:49:45,900
YOU KNOW CONTRIBUTE SOME STONES

1281
00:49:45,900 --> 00:49:48,103
TO THAT BIG MOSAIC WITH THE

1282
00:49:48,103 --> 00:49:49,304
RESEARCH COMMUNITY, IT'S A VERY

1283
00:49:49,304 --> 00:49:51,606
EXCITING THING TO DO.

1284
00:49:51,606 --> 00:49:54,242
AND AGAIN, SO THIS WORK IS DONE

1285
00:49:54,242 --> 00:49:57,178
BY ABSOLUTELY AMAZING PEOPLE IN

1286
00:49:57,178 --> 00:49:57,912
MY LAB.

1287
00:49:57,912 --> 00:49:59,714
FORMER MEMBERS OF THE LAB AND

1288
00:49:59,714 --> 00:50:02,083
COLLABORATORS REALLY ALL OVER

1289
00:50:02,083 --> 00:50:07,956
THE WORLD AND OBVIOUSLY AMAZING

1290
00:50:07,956 --> 00:50:09,924
FUNDING, FUNDING SOURCES AND

1291
00:50:09,924 --> 00:50:11,326
THEN, IT'S ALWAYS IMPORTANT TO

1292
00:50:11,326 --> 00:50:13,995
HAVE FUN IN THE LAB, THIS IS

1293
00:50:13,995 --> 00:50:15,163
[INDISCERNIBLE] PROSENTING A

1294
00:50:15,163 --> 00:50:16,998
POSTER AT THE MOILOID MEETING

1295
00:50:16,998 --> 00:50:21,603
THIS IS HANNAH [INDISCERNIBLE]

1296
00:50:21,603 --> 00:50:22,804
AND JULIA, AND THE

1297
00:50:22,804 --> 00:50:26,074
[INDISCERNIBLE] MEETING THIS WAS

1298
00:50:26,074 --> 00:50:28,376
A DESSERT HE DESIGNED FOR THE

1299
00:50:28,376 --> 00:50:29,911
MEETING AND I CAN'T TELL YOU HOW

1300
00:50:29,911 --> 00:50:32,113
IT TASTED, I DIDN'T GET TO HAVE

1301
00:50:32,113 --> 00:50:34,983
IT, AND NEW HAVEN HAS GOOD PIZZA

1302
00:50:34,983 --> 00:50:45,527
AND SO THANK YOU FOR BEING HERE

1303
00:50:50,732 --> 00:50:50,865
TODAY.

1304
00:50:50,865 --> 00:50:52,000
[ APPLAUSE ]

1305
00:50:52,000 --> 00:50:53,201
>> OKAY, THAT WAS GREAT.

1306
00:50:53,201 --> 00:50:54,702
THE MOUSE MODEL WAS REALLY

1307
00:50:54,702 --> 00:50:56,905
EXCITING, I HAVE A QUESTION

1308
00:50:56,905 --> 00:50:58,439
ABOUT THE SPLICING FACTOR

1309
00:50:58,439 --> 00:50:58,706
MUTATIONS.

1310
00:50:58,706 --> 00:51:00,308
I THINK WHAT'S STRIKING TO ME IS

1311
00:51:00,308 --> 00:51:02,343
THE FACT THAT THERAPY AND

1312
00:51:02,343 --> 00:51:05,280
HETEROZYGOTE AND YET, THE MUTANT

1313
00:51:05,280 --> 00:51:07,715
PROTEIN HAS AN OUTSIZED ROLE, OR

1314
00:51:07,715 --> 00:51:08,116
OUTSIZED FUNCTION.

1315
00:51:08,116 --> 00:51:10,885
DO YOU THINK THAT THAT'S

1316
00:51:10,885 --> 00:51:12,287
BECAUSE--THESE PROTEINS TEND TO

1317
00:51:12,287 --> 00:51:14,022
BE HIGHLY EXPRESSED, DO YOU

1318
00:51:14,022 --> 00:51:16,524
THINK IT'S BECAUSE OF A FUNCTION

1319
00:51:16,524 --> 00:51:19,194
OF A MUTANT PROINE, OR LESS

1320
00:51:19,194 --> 00:51:20,795
WILD-TYPE PROTEIN HAS BEEN MADE.

1321
00:51:20,795 --> 00:51:23,364
HAVE YOU OR ANYBODILE CHECKED IF

1322
00:51:23,364 --> 00:51:25,466
YOU INHIBIT THE MUTANT PROTEIN

1323
00:51:25,466 --> 00:51:30,038
DO YOU GET A RESTORATION OF

1324
00:51:30,038 --> 00:51:31,506
FUNCTION?

1325
00:51:31,506 --> 00:51:34,776
>> OKAY, SO MICE THAT ARE

1326
00:51:34,776 --> 00:51:35,910
HETEROZYGOUS FOR SHE'S SPLICING

1327
00:51:35,910 --> 00:51:36,878
FACTORS ARE DOING PRETTY WELL,

1328
00:51:36,878 --> 00:51:38,513
SO THEY DON'T HAVE MUCH OF A

1329
00:51:38,513 --> 00:51:44,185
PHENOTYPE SO I THINK THAT'S 1

1330
00:51:44,185 --> 00:51:44,419
ASPECT.

1331
00:51:44,419 --> 00:51:49,157
YOU KNOW 1 OF THE REASONS WE

1332
00:51:49,157 --> 00:51:51,025
WERE EXCITED THAT MUTANT 1

1333
00:51:51,025 --> 00:51:54,128
ACTUALLY BINDS THE RNA IS MAYBE

1334
00:51:54,128 --> 00:51:55,196
NOW AND DIFFERENTLY MAYBE NOW

1335
00:51:55,196 --> 00:51:58,700
THERE'S SOMETHING YOU COULD

1336
00:51:58,700 --> 00:51:58,967
INHIBIT,.

1337
00:51:58,967 --> 00:52:00,735
>> WITH THE DRUG
DEVELOPERS, I

1338
00:52:00,735 --> 00:52:01,736
WOULDN'T KNOW WHERE TO START TO

1339
00:52:01,736 --> 00:52:04,973
TRY TO DO THAT.

1340
00:52:04,973 --> 00:52:07,375
BUT I THINK, SO, IN THE CELL

1341
00:52:07,375 --> 00:52:12,313
LINES YOU KNOW WE EXOGLY EXPRESS

1342
00:52:12,313 --> 00:52:13,982
THESE PROTEINS, AND IT'S AULENTY

1343
00:52:13,982 --> 00:52:16,084
VIRAL VECTOR SO WE TRY TO

1344
00:52:16,084 --> 00:52:17,452
TRANSDUCE THESE CELLS AND IT WAS

1345
00:52:17,452 --> 00:52:21,289
1 SO WE GET 1 INTERNATIONAL

1346
00:52:21,289 --> 00:52:22,891
CLASSIFICATION GRAIN BUT THEN

1347
00:52:22,891 --> 00:52:25,860
IT'S STILL AULENTY VIRAL LTR AND

1348
00:52:25,860 --> 00:52:28,496
SO FOR THE SFR2, THERE'S RNA,

1349
00:52:28,496 --> 00:52:31,132
THERE'S LOTS OF MUSEUM TABT SSF2

1350
00:52:31,132 --> 00:52:33,001
BUT AT THE PROTEIN LEVEL THERE

1351
00:52:33,001 --> 00:52:35,036
ISN'T THAT MUCH MORE.

1352
00:52:35,036 --> 00:52:37,272
I THINK THE MOST WE GET

1353
00:52:37,272 --> 00:52:39,674
OVEREXPRESSED AT THE PROTEIN

1354
00:52:39,674 --> 00:52:43,344
LEVEL IS 1.5 X OR SO, SO I THINK

1355
00:52:43,344 --> 00:52:45,213
THERE'S A VERY, VERY TIGHT

1356
00:52:45,213 --> 00:52:47,682
REGULATION OF STILL SPLICING

1357
00:52:47,682 --> 00:52:49,384
FACTOR STOICHIOMETRY, AND

1358
00:52:49,384 --> 00:52:52,120
SO,--SO I THINK IT'S A GAME, I

1359
00:52:52,120 --> 00:52:56,891
THINK IT'S A GAIN OF FUNCTION

1360
00:52:56,891 --> 00:52:57,825
AND NOT NECESSARILY--BUT IT

1361
00:52:57,825 --> 00:53:00,194
COULD ALSO BE A COMBINATION.

1362
00:53:00,194 --> 00:53:01,796
>> SO HAS ANYONE TRIED TO

1363
00:53:01,796 --> 00:53:03,865
OVEREXPRESS THE WILD-TYPE

1364
00:53:03,865 --> 00:53:04,098
PROTEIN?

1365
00:53:04,098 --> 00:53:09,504
>> SO ALL OUR WORK IN CELL
LINES

1366
00:53:09,504 --> 00:53:11,739
WE OVEREXPRESS WILD-TYPE MUTANT

1367
00:53:11,739 --> 00:53:13,708
SEPARATELY, WE HAVE NOT DONE

1368
00:53:13,708 --> 00:53:21,115
THAT TOGETHER. 

1369
00:53:21,115 --> 00:53:21,716
>> OKAY, THANKS.

1370
00:53:21,716 --> 00:53:23,384
>> HAY, THANK YOU SO MUCH,
THAT

1371
00:53:23,384 --> 00:53:24,385
WAS FASCINATING.

1372
00:53:24,385 --> 00:53:26,054
I HAVE A QUESTION ABOUT STRESS

1373
00:53:26,054 --> 00:53:28,489
GRANULES, SO WHEN YOU PULL DOWN

1374
00:53:28,489 --> 00:53:30,692
STRESS GRANULES AND WHICH RNAs

1375
00:53:30,692 --> 00:53:32,026
WERE UPOR COUNSEL REGULATED AND

1376
00:53:32,026 --> 00:53:34,696
I WONDERED IF YOU LOOK TO SEE A

1377
00:53:34,696 --> 00:53:35,863
CORRELATION FOR STRESS GRANULES

1378
00:53:35,863 --> 00:53:37,665
THAT WERE DOWN REGULATED IF THEY

1379
00:53:37,665 --> 00:53:39,600
WERE DISPLACED DRCH LITE AND

1380
00:53:39,600 --> 00:53:41,269
BECAME SHORTER SINCE THE LEPGHT

1381
00:53:41,269 --> 00:53:43,471
OF RNAs TEND TO CORRELATE HOW

1382
00:53:43,471 --> 00:53:44,973
ENRICHED OR NOT ENRICHED THEY

1383
00:53:44,973 --> 00:53:47,041
ARE IN THE STRESS GRANULES.

1384
00:53:47,041 --> 00:53:49,143
NYEAH, THAT'S WHATEE JULIA'S

1385
00:53:49,143 --> 00:53:51,012
LOOKING AT NOW SHE WAS JUST AT

1386
00:53:51,012 --> 00:53:52,981
THE RNA SOCIETY MEETING THAT'S

1387
00:53:52,981 --> 00:53:54,415
WHAT SHE CRANKED THIS DATA FOR,

1388
00:53:54,415 --> 00:53:55,950
I'VE SEEN IT LIKE 5 MINUTES SO

1389
00:53:55,950 --> 00:53:57,618
WE DON'T HAVE THAT YET, BUT

1390
00:53:57,618 --> 00:53:59,520
YEAH, YEAH, YOU'RE RIGHT, SO

1391
00:53:59,520 --> 00:54:00,355
THAT'S FIRST YOU WILL HAVE TO

1392
00:54:00,355 --> 00:54:01,990
LOOK AT AND YOU KNOW IT'S A

1393
00:54:01,990 --> 00:54:03,624
LITTLE BIT SCARY BECAUSE WE MAY

1394
00:54:03,624 --> 00:54:07,362
OR MAY NOT FIND SOMETHING.

1395
00:54:07,362 --> 00:54:13,534
>> THANK YOU. 

1396
00:54:13,534 --> 00:54:14,836
>> REALLY GOOD QUESTION.

1397
00:54:14,836 --> 00:54:15,937
>> GREAT TALK.

1398
00:54:15,937 --> 00:54:21,409
SO IT'S EXCITING TO SEE THIS IS

1399
00:54:21,409 --> 00:54:22,043
[INDISCERNIBLE] STRESS GRANULES

1400
00:54:22,043 --> 00:54:28,082
AND SO, WHEN YOU LOOK AT THE

1401
00:54:28,082 --> 00:54:30,218
OVERLAP AFFECTED VENTS BETWEEN

1402
00:54:30,218 --> 00:54:30,885
[INDISCERNIBLE] MUTATIONS, THIS

1403
00:54:30,885 --> 00:54:33,021
IS GOING ON THIS WAY THINKING

1404
00:54:33,021 --> 00:54:34,389
INDEPENDENTLY, THIS MUTATIONS

1405
00:54:34,389 --> 00:54:36,457
MAY AFFECT DIFFERENT

1406
00:54:36,457 --> 00:54:37,725
TRANSCRIPTS,.

1407
00:54:37,725 --> 00:54:39,961
>> YEAH, YEAH, GOOD
QUESTION.

1408
00:54:39,961 --> 00:54:44,165
SO WE HAVEN'T ANALYZED PATIENT

1409
00:54:44,165 --> 00:54:46,667
DATA SETS BASED ON THAT.

1410
00:54:46,667 --> 00:54:49,570
YOU KNOW IN OUR--IF I REMEMBER

1411
00:54:49,570 --> 00:54:51,873
CORRECTLY IN OUR--BECAUSE OUR

1412
00:54:51,873 --> 00:54:54,175
SRF2 KIND OF LIKE SPLICING

1413
00:54:54,175 --> 00:54:56,744
ANALYSIS DATA WAS A WHILE AGO,

1414
00:54:56,744 --> 00:54:59,013
THERE WEREN'T ACTUALLY A TON OF

1415
00:54:59,013 --> 00:55:01,783
EVENTS SO I THINK NOW, THAT'S

1416
00:55:01,783 --> 00:55:04,118
WHAT JULIA WILL HAVE TO DO IS

1417
00:55:04,118 --> 00:55:06,988
REALLY, SO WE ACTUALLY BACK THEN

1418
00:55:06,988 --> 00:55:08,623
WE PUBLISHED SRF 2 TO

1419
00:55:08,623 --> 00:55:10,091
[INDISCERNIBLE] CLIP AND THAT'S

1420
00:55:10,091 --> 00:55:13,895
NOT SUCH A GREAT TECHNIQUE SO WE

1421
00:55:13,895 --> 00:55:15,296
ACTUALLY HAVE SSF2 ECLIP DATA

1422
00:55:15,296 --> 00:55:16,964
FROM LIKE 4 YEARS AGO THAT WE

1423
00:55:16,964 --> 00:55:18,833
HAVEN'T ANALYZED WHICH IS

1424
00:55:18,833 --> 00:55:24,338
EMBARRASSING BUT NOW WE'RE DOING

1425
00:55:24,338 --> 00:55:24,572
THAT.

1426
00:55:24,572 --> 00:55:26,207
I CAN'T AT ALL, BUT I THINK IT

1427
00:55:26,207 --> 00:55:27,775
WILL BE A REALLY INTERESTING.

1428
00:55:27,775 --> 00:55:30,011
SO THIS--YOU KNOW THE STRESS

1429
00:55:30,011 --> 00:55:31,179
GRANULES SIGNATURE, RIGHT, SHE'S

1430
00:55:31,179 --> 00:55:32,647
TAKING, SHE TOOK ALL THESE DATA

1431
00:55:32,647 --> 00:55:34,382
SETS, AND I THINK SHE'S PUTTING

1432
00:55:34,382 --> 00:55:35,683
THAT TOGETHER AS A LITTLE THING

1433
00:55:35,683 --> 00:55:38,553
FOR THE RNA SOCIETY, RIGHT, SO

1434
00:55:38,553 --> 00:55:39,587
YOU PULLED OUT ALL THE OTHER

1435
00:55:39,587 --> 00:55:41,022
REFRESH YOUR RECOLLECTION NAs

1436
00:55:41,022 --> 00:55:43,658
AND PROTEINS THAT HAVE BEEN

1437
00:55:43,658 --> 00:55:47,728
ISOLATED AND STRESS GRANULES, BY

1438
00:55:47,728 --> 00:55:49,864
VARIOUS GROUPS AND A LOT OF THAT

1439
00:55:49,864 --> 00:55:51,466
FOR EXAMPLE, I THINK LIKE

1440
00:55:51,466 --> 00:55:52,767
[INDISCERNIBLE] AND 2 AND 3

1441
00:55:52,767 --> 00:55:56,003
CELLS SO THERE'S STILL MORE THAT

1442
00:55:56,003 --> 00:55:57,271
CAN BE DONE.

1443
00:55:57,271 --> 00:55:58,239
HER STRESS GRANULE SIGNATURE,

1444
00:55:58,239 --> 00:56:01,242
SHE HAS TO NOW LOOK AT THAT

1445
00:56:01,242 --> 00:56:02,610
SPECIFICALLY FOR SSRF 2, BECAUSE

1446
00:56:02,610 --> 00:56:04,946
A LOT IS TILL INFLUENCED BY HER

1447
00:56:04,946 --> 00:56:06,581
DATA JUST BECAUSE SHE DOESN'T

1448
00:56:06,581 --> 00:56:09,417
HAVE THE BINDING, SPLICING,

1449
00:56:09,417 --> 00:56:09,617
YEAH.

1450
00:56:09,617 --> 00:56:10,284
YEAH, I DON'T KNOW.

1451
00:56:10,284 --> 00:56:11,986
I THINK IT WOULD BE NICE IF

1452
00:56:11,986 --> 00:56:14,422
THERE WERE A COMMON MECHANISM,

1453
00:56:14,422 --> 00:56:15,823
BECAUSE IT'S INTERESTING, RIGHT

1454
00:56:15,823 --> 00:56:19,760
BECAUSE WE HAVE PAPERS

1455
00:56:19,760 --> 00:56:22,530
SUGGESTING THAT SSF2 UGF 1 FOR

1456
00:56:22,530 --> 00:56:24,932
EXAMPLE, ALSO RESIDE IN MORE

1457
00:56:24,932 --> 00:56:29,003
R-LOOPS, BUT EVEN THE MUTATIONS

1458
00:56:29,003 --> 00:56:29,804
FOR [INDISCERNIBLE], THEY'RE NOT

1459
00:56:29,804 --> 00:56:33,708
THE SAME, THEY CURVE, THEY HAVE

1460
00:56:33,708 --> 00:56:34,909
DIFFERENT PHENOTYPES, THEY DO

1461
00:56:34,909 --> 00:56:36,544
DIFFERENT THINGS, SO WHAT'S

1462
00:56:36,544 --> 00:56:37,345
COMMON BETWEEN THOSE 2?

1463
00:56:37,345 --> 00:56:43,584
I DON'T THINK WE KNOW.

1464
00:56:43,584 --> 00:56:47,088
>> THANKS. 

1465
00:56:47,088 --> 00:56:49,757
>> ANY OTHER QUESTIONS?

1466
00:56:49,757 --> 00:56:50,124
ANYBODY ONLINE?

1467
00:56:50,124 --> 00:56:53,427
I HAVE NO IDEA HOW THIS WORKS.

1468
00:56:53,427 --> 00:56:55,763
OKAY, SO, IF THERE ARE NO

1469
00:56:55,763 --> 00:56:57,265
FURTHER QUESTIONS, LET'S THANK

1470
00:56:57,265 --> 00:56:59,634
STEPHANIE FOR A GREAT

1471
00:56:59,634 --> 00:56:59,967
PRESENTATION.

1472
00:56:59,967 --> 00:57:10,178
[ APPLAUSE ]