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Nuclear damage and miscounted chromosomes: Human T cell leukemia virus transformation of cells

February 18, 2020


>>>SO I’D LIKE TO WELCOME YOU TO THE ANNUAL GEORGE KHOURY LECTURE AT THE NIH. IT’S MY GREAT PLEASURE TODAY TO ACTUALLY INTRODUCE TWO OUTSTANDING NIH RESEARCHERS. THE FIRST, OF COURSE, IS GEORGE KHOURY, FOR WHOM THIS LECTURE IS NAMED AND WHO WAS INCREDIBLE SCIENTIST AT THE NIH. UNFORTUNATELY DIED 25 YEARS AGO FROM LYMPHOMA, AND WE HAVE HAD THIS LECTURE VIRTUALLY EVERY YEAR SINCE THEN IN HIS HONOR. ANYONE WHOEVER MET GENERAL ANYHOW THAT HE WAS GOING TO BE SUCCESSFUL — GEORGE — I NOTICED MARILYN KHOURY IS IN THE AUDIENCE AND SHE WILL ASSURE US THAT IS THE CASE. HE WAS AN HONORS GRADUATE OF PRINCETON AND HARVARD MEDICAL SCHOOL. AND THEN AT THE RIPE OLD AGE OF 33, BECAME CHIEF OF THE NCI VIRUS TUMORS SECTION AT NIH AND NCI. IN HIS ALL-TOO SHORT CAREER AT THE NIH, GEORGE MANAGED TO CONTRIBUTE IN AN INCREDIBLE WAY TO SCIENCE HERE, PUBLISHING OVER 140 ORIGINAL RESEARCH PAPERS AND EARNING MEMBERSHIP IN THE NATIONAL ACADEMY OF SCIENCES, WHICH IS AWARDED JUST A FEW MONTHS BEFORE HIS DEATH. HIS PRIMARY INTEREST WAS IN REGULATION OF GENE EXPRESSION IN MAMMALIAN CELLS. WE ALL KNOW ABOUT THE IMPORTANCE OF SEQUENCES WE CALL ENHANCERS, BUT PROBABLY SOME PEOPLE IN THE AUDIENCE DON’T REALIZE THAT THE TERM WAS OR AND THE CONCEPT WAS INVENTED BY GEORGE COREY. AND THAT IS A LASTING LEGACY FOR WHICH WE ARE ALL RATHER GRATEFUL. HE MADE MANY SEMINOLE CONTRIBUTIONS TO THE FIELD OF VIRALLY. THERE WASN’T MUCH KNOWN ABOUT HOW VIRUSES CAUSE CANCER WHEN HE STARTED HIS WORK IN THE 1970s. HE WAS AMONG THE FIRST PEOPLE TO REALIZE THE WAY IN WHICH SMALL VIRUSES, SUCH AS SV40, WERE ABLE TO CAUSE CANCER BY TURNING ON GENE EXPRESSION AND SUB VERTING NORMAL GROWTH MECHANISMS PRESENT IN MAMMALIAN CELLS. HIS SCIENTIFIC PROWES WAS ONLY MATCHED BY HIS INCREDIBLE MENTORING. IN THAT SHORT PERIOD OF TIME, A HUGE NUMBER OF REALLY SUCCESSFUL SCIENTISTS PASSED THROUGH HIS LABORATORY AND EACH ONE OF THEM BENEFITED FROM HIS INDIVIDUAL CARE AND ATTENTION AND MANY OF US WHO WERE FRIENDS AND COLLEAGUES ALSO BENEFITED FROM HIS SAGE ADVICE, HIS WISDOM. HE REALLY LOVED SCIENCE. HE LOVED TO GO TO SEMINARS, AND WOULD GET DEEPLY ENGAGED IN ANY SCIENTIFIC SNUG A WHOLE VARIETY OF ISSUES. THE KIND OF PERSON WHO REALLY ADDED PIZAZZ TO THE NIH INTEREST MURAL PROGRAM AND WE REALLY, REALLY MISS HIM. SO IN HIS HONOR, WE HAVE CREATED THE GEORGE COREY LECTURE SERIES. IT’S ONE OF OUR WEDNESDAY AFTERNOON LECTURES. AND WE EXPECT THAT EVERY YEAR, A LEADING INVESTIGATOR, ESPECIALLY PEOPLE WHO HAVE BEEN ASSOCIATED OR HAD BEEN ASSOCIATED WITH GEORGE IN HIS CAREER, WOULD BE ONE OF THE SPEAKERS. I SHOULD POINT OUT THAT THERE IS ANOTHER GEORGE COREY MEMORIAL LECTURE SERIES AT THE WISTAR INSTITUTE, AND I HAD THE PRIVILEGE LAST YEAR TO GIVE THAT LECTURE WHICH I WAS DEEPLY HONORED AND IT MADE ME APPRECIATE THAT GENERAL’S IMPACT EXTENDED WAY BEYOND THE NIH. PEOPLE AND MANY INSTITUTIONS AT PRINCETON, THE WISTAR INSTITUTE WHICH HE WAS ENGAGED IN OVER THE YEARS, WHO REMEMBER HIM EXTREMELY FONDLY AND WANTED ALSO TO MEMORIALIZE HIM WITH A LECTURE SERIES. SO, THE SECOND OUTSTANDING NIH INTRAMURAL SCIENTIST I’LL TALK ABOUT IT IS — STUDY OUR SPEAKER. THAT KUAN-TEH JEANG. ONE OF THE QUOTE, YOUNG SCIENTISTS, WHOM GEORGE COREY HELPED TO TRAIN. YOU NEED A BACKSTORY HERE. EVERY YEAR WE HELP TO CHOOSE THE SPEAKER FOR THE ELEC TER. DIANE GRICH FROM HOPKINS, FRANK RASHER FROM PENN, JUST IN THE LAST THREE YEARS. SO, HAS DONE A TERRIFIC JOB AND WITH LARRY BRODIE, I’M SORRY, JOHN BRADY WHO DIED UNFORTUNATELY AT AN EARLY AGE A FEW YEARS AGO, THEY REALLY RAN THIS LECTURE SERIES FOR MANY YEARS. SO THIS YEAR, AS I WAS FRETTING OVER WHO WOULD BE THE BEST SPEAKER, I HAD AN EPIPHANY AND WHAT STRUCK ME WAS THAT TEH, HIMSELF, A REALLY TRULY OUTSTANDING SCIENTIST, WOULD BE A TERRIFIC SPEAKER IN THIS SERIES. WE INVITED HIM. HE DITHERED A LITTLE BIT ABOUT WHETHER HE REALLY WANTED TO DO IT, BUT HE DID AGREE AND WE ARE DELIGHTED TO HAVE HIM HERE TODAY. SO TEH, MANY KNOW, IS A REAL DYNAMO. HE HAS BEEN A SCIENTIST AT NIAID FOR 25 YEARS. WHERE HE IS NOW CHIEF OF THE MOLECULAR VIROLOGY SECONDS OF THE LABORATORY OF MOLECULAR MICROBIOLOGY, PUBLISHED 250 SCIENTIFIC PAPERS AND #EDITED SIX BOOKS. HE IS THE EDITOR-IN-CHIEF OF RETRO VIROLOGY AND EDITOR OF CELL AND BIOSCIENCE AND ASSOCIATE EDITOR FOR CANCER RESEARCH AND SERVES ON EDITORIAL BOARDS FROM THE JVC AND JOURNAL OF VIROLOGY. HE IS ALSO PRESIDENT OF THE SOCIETY OF CHINESE BIOSCIENTISTS IN AMERICA. AND IN THAT CAPACITY, HAS BEEN OUTSPOKEN IN HIS DESIRE FOR INCREASED REPRESENTATION AND LEADERSHIP POSITIONS FOR ASIAN-AMERICAN SCIENTISTS AND HE REALLY HAS BEEN VERY COMPELLING IN THAT POSITION AND HOPEFULLY WILL BE EXTREMELY SUCCESSFUL IN ADVANCING CAREERS OF VARIED PEOPLE WHO REALLY DESERVE RECOGNITION. SO, SOME OF HIS MORE RECENT WORK INCLUDES FINDINGS ON GENOME-WIDE CHARACTERIZATION OF 252 HUMAN HOST CELL FACTORS IN GER KET T-CELLS CONTRIBUTORY TO HIV1 REPLICATION, AND CHARACTERIZATION OF SEVERAL ONCOGENIC MICRORNS IN THEIR CONTRIBUTION TO HLV1 ONCOGENESIS AND THE LIST OF HIS CONTRIBUTIONS IS ALMOST ENDLESS. HE WAS AN UNDERGRADUATE AT MIT AND GOT HIS MD AND Ph.D. FROM HOPKINS UNIVERSITY SCHOOL OF MEDICINE. SO LET’S WELCOME KUAN-TEH JEANG WHOSE TALK IS NUCLEAR DAMAGE AND ANEUPLOIDY HTLV1 AND LOU KEIM GENESIS. THANK YOU.>>THANK YOU, MICHAEL T IS INDEED A GREAT HONOR TO BE HERE TO BOTH GIVE THE LECTURE AND ALSO TO CELEBRATE GEORGE COREY AND HIS CONTRIBUTIONS TO SCIENCE. AS MICHAEL MENTIONS,ED I WAS ONE OF GEORGE’S LAST POSTDOCTORAL FELLOWS, AND I’M REALLY INDEBTED TO GEORGE IN THE SENSE THAT I LEARNED A LOT ABOUT SCIENCE FROM HIM AND PERHAPS EACH MORE ABOUT BEING A HUMAN BEING, AND HOW TO SORT OF CONDUCT AND TEACH SCIENCE. SO, SOME OF YOU HAVE VISITED MY OFFICE OVER THE YEARS. AND I THINK YOU HAVE ALWAYS NOTICED, AND ESPECIALLY NEW POSTDOCTORAL FELLOWS WHO COME INTO THE LAB, YOU ALWAYS NOTICED THIS PICTURE HANGING IN MY OFFICE. AND IT HAS BEEN THERE FOR THE LAST 25 YEARS. AND SOME PEOPLE WONDER WHO IS THIS PERSON? AND THIS IS ACTUALLY THE PICTURE THAT GEORGE GAVE TO ME A FEW WEEKS BEFORE HE PASSED AWAY. AND IT CERTAINLY IS A PICTURE THAT I HAVE TREASURED AND HAVE HUNG IT IN MY OFFICE FOR THE LAST 25 YEARS. NOW, I WOULD LIKE TO SHARE A COUPLE OF OTHER PICTURES WITH YOU. THESE ARE PICTURES OF GEORGE AND A LITTLE BIT EARLIER, BEFORE WHEN HE SORT OF STARTED LOOKING SORT OF WAYNE FROM THE DISEASE. AND SO, THIS IS A VERY NICE PICTURE OF GEORGE AND MARILYN. THESE ARE PICTURES OF A PARTY THAT WE HAD AT GEORGE AND MARILYN’S HOUSE DURING THE PERIOD WHEN GEORGE WAS DOING WELL IN HIS REMISSION. AND YOU CAN SEE SOME INDIVIDUALS, I THINK SOME OF THEM ARE STILL HERE AT THE NIH. THIS IS LIONEL WHO IS OVER THERE AT NCI FREDRICK. I BELIEVE THIS IS JEFF GREEN. IT’S REMARKABLE HOW SOME OF THESE PEOPLE HAVE CHANGED AND THEY LOOK SO MUCH BETTER WHEN THEY WERE YOUNGER. [ LAUGHTER ] SOME OF YOU KNOW CAMEL WHO IS NOW AT JEFFERSON. THIS WAS CAMEL WHEN HE WAS YOUNGER. THIS WAS OF COURSE GEORGE, AND THIS IS ACTUALLY AN IMAGE OF ME SOME 30 YEARS AGO. AND THIS IS LOU. AND THIS LOVELY 20 SOMETHING-YEAR-OLD GIRL SITTING THERE IS ACTUALLY MY WIFE, DIANE, WHO ALSO CAME TO THE PARTY. AND HERE ARE A COUPLE OF — HERE IS A PICTURE OF GENERAL WITH NADIA ROSENTHAL, WHICH I KNOW MANY OF YOU ARE FRIENDS WITH. NADIA WAS ALSO HERE AT THE NIH MANY YEARS AGO. SO, THESE ARE SOME OF THE SORT OF OFF-HOUR GOOD TIMES THAT WE HAD WITH GEORGE. AND GEORGE WAS A TERRIFIC SCIENTIST AND I THINK ALSO A TERRIFIC MENTOR AND TEACHER TO US. SO, WITH THAT THEN, SHORTLY AFTER GEORGE PASSED AWAY, YOU KNOW, I HAD THE DESIRE THAT BECAUSE I FELT I HAD BENEFITED SO MUCH FROM GEORGE, THAT I SHOULD DO SOMETHING TO TRY TO REMEMBER GEORGE. AND SO, IN 1994. THIS WAS THE VERY FIRST POSTER OF THE VERY FIRST LECTURE, ARMY LEVINE, A GREAT FRIEND OF GEORGE’S WHO CAME AND TALKED ABOUT p53 TUMOR SUPPRESSOR, A TOPIC THAT IS VERY NEAR AND CLOSE OR NEAR AND DEAR TO GEORGE’S HEART. AND AS YOU CAN SEE FROM THOSE EARLY DAYS WHEN WE SORT OF WERE ON A LOW BUDGET, WE ACTUALLY ENDED UP HAVING — YOU MAY NOT BE ABLE TO READ THESE NAMES BUT THESE WERE A NUMBER OF GEORGE’S POSTDOCTORAL FELLOWS WHO PERSONALLY CONTRIBUTED MONEY TO BOOTSTRAP THIS VERY 50 LECTURE, INCLUDING JOE JOHN, WHO I KNOW IS IN THE AUDIENCE TODAY. AND SO, YOU CAN APPRECIATE THAT BACK IN THOSE DAYS WHEN YOU FIRST STARTED LECTURE WITH LOW BUDGET, THAT YOU SORT OF PRINT THE PROGRAM AND THE ANNOUNCEMENT ON THIS SORT OF BROWN BAG PAPER. SO THAT’S REALLY INDICATING THAT THIS IS SORT OF A — THE EARLY DAYS. BUT THEN BY-AND-BY, WE SORT OF GOT LUCKY BECAUSE WHEN MICHAEL BECAME THE INTRAMURAL DIRECTOR, WE WERE ABLE TO HAVE THE LECTURE BECOME PART OF THE WEDNESDAY AFTERNOON LECTURE SERIES. AND THEN YOU CAN SEE THAT WE GOT MUCH NICER POSTER DESIGNS AND BETTER FRAMING AND GOT A NUMBER OF ADDITIONAL LOOM NARRIES COME TO GIVE THE LECTURE — LUMINAIRES. AND OVER THE 18 YEAR HISTORY OF THE GEORGE KHOURY LECTURE, YOU CAN SEE WE HAVE REALLY HAD A NUMBER OF ALL-STAR THAT IS HAVE COME. THESE ARE FRIENDS, COLLEAGUES, STUDENTS OF GEORGE’S, THAT HAVE COME AND REALLY HAVE PRESENTED WONDERFUL SCIENTIFIC TALKS AND ALSO IN MANY CASES, REALLY, REALLY PERSONAL STORIES ABOUT GEORGE. AND SO HERE IS A PICTURE OF MY FRIEND AND THE FORMER GEORGE KHOURY LECTURER IN 2009, FRANK RAUSCHER AND THIS IS A VERY NICE PICTURE OF GEORGE HE SHOWED DURING HIS LECTURE. NOW I WANT TO SAY WITH EVERY SUCCESSFUL LECTURE SERIES, THERE IS ALWAYS A COMMUNITY OF INDIVIDUALS — IT CERTAINLY IS NOT ME WE OWE CREDIT TO. AND IN THE CASE OF THE GEORGE KHOURY LECTURE, THERE ARE A NUMBER OF INDIVIDUALS THAT WE SHOULD CREDIT, WHICH IS THAT INITIALLY DURING THE EARLY DAYS, ALAN RAPS EN, KIRSCHSTEIN WAS VERY INSTRUMENTAL IN HELPING ME SET UP THE LECTURE AND OVER THE COURSE, WE ALWAYS HAD VERY WONDERFUL SUPPORT FROM MARILYN AND WE ALSO HAD SUZANNE, MICHAEL AND IRA FORM THE COMMUNITY OF INDIVIDUALS WHO HAVE REALLY HELPED US GET THE LECTURE GOING AND CONTINUING THE LECTURE IN THE HIGH-QUALITY WAY. WE HAVE ALSO HAD BERNIE, TOM AND BRUCE AND DOUG AND WARREN LEONARD AND MALCOLM ARNOLD. SO THESE ARE THE INDIVIDUALS THAT REALLY THAT WE OWE CREDIT TO OVER THE LAST 18 YEARS FOR CONTINUING THIS LECTURE AND REALLY MAINTAINING THE HIGH QUALITY OF THE LECTURE. BUT OUT OF ALL OF THESE PEOPLE, THE ONE PERSON THAT I REALLY WANT TO ACKNOWLEDGE PERHAPS 234 AN OUTSTANDING WAY IS OF COURSE, MY FRIEND AND FORMER COLLEAGUE IN GEORGE’S LAB, JOHN BRADY. JOHN AS MICHAEL MENTIONED, PASSED AWAY A FEW YEARS AGO. AND DURING THE COURSE OF THOSE 18 YEARS, THERE WAS A PERIOD OF TIME WHEN I WAS ACTIVE LOOLY NEGOTIATING FOR A POSITION ELSEWHERE. AND DURING THAT TIME, JOHN STEPPED IN FOR SEVERAL YEARS, BECAUSE I WAS CONCERNED THAT I NEEDED SOMEONE THAT COULD REALLY CONTINUE AND DO THE GEORGE KHOURY LECTURE IN A REALLY EFFICIENT AND OUTSTANDING FASHION. AND SO, THERE WERE A LARGE NUMBER OF YEARS THERE IN THE MIDDLE WHERE JOHN STEPPED IN AND DID THE BULK OF THE WORK, MAKING SURE THAT THE LECTURE WAS DONE IN A REALLY OUTSTANDING FASHION. AND I ALSO OWE JOHN A LOT BECAUSE WHEN I ARRIVED IN THE LAB, JOHN WAS ALREADY THERE AND WAS SORT OF THE SENIOR RESEARCHER. IN MANY WAYS, I ALSO LEARNED A LOT FROM JOHN SCIENTIFICALLY. WE WILL MISS JOHN AND I VALUE HIS MEMORIES AS I ALWAYS HAVE VALUES GENERAL’S MEMORIES. SO, LET ME MOVE ON THEN IT TALK ABOUT SCIENCE. AND HERE, AS MANY OF YOU KNOW MY LABORATORY WORKS ON TWO DIFFERENT RETT REVIRUSES. TWO DIFFERENT HUMAN RETROVIRUSES. HTLV1 WHICH I STARTED IN GEORGE’S LAB. AND HIV1 WHICH I CONTINUED AFTER LEAVING GEORGE’S LAB AND JOINING MALCOLM MARTYN’S ORGANIZATION. SO, WHEN I WAS ASKED TO SPEAK AND GIVE THIS YEAR’S GEORGE KHOURY LECTURE, FOR PERSONAL REASONS, AND ALSO SCIENTIFIC REASONS, ELECTED OF COURSE TO TALK ABOUT HTLV1, BECAUSE THIS IS INDEED THE VIRUS THAT I STARTED WORKING AND THE VIRUS THAT GEORGE TAUGHT ME ABOUT. SO, I THOUGHT THIS WOULD BE THE MORE IMPORTANT, MORE APPROPRIATE TOPIC TO SORT OF DEVOTE THIS YEAR’S LECTURE ON. BUT NEVERTHELESS, I WANT TO EMPHASIZE TO YOU THAT IN A LARGE PART, MY CAREER STARTED IN HIV SORT OF USING MANY OF THE IDEAS AND MANY OF THE TEACHINGS THAT I ACTUALLY LEARNED FROM GENERAL WHEN I WAS WORKING ON HTLV1. SO, AS MICHAEL ELUDED TO, GEORGE WAS INSTRUMENTAL IN THE DISCOVERY OF THE CONCEPT OF ENHANCER SEQUENCES. SO THIS WAS A REVIEW ARTICLE THAT HE AND I WROTE TOGETHER ALL THE WAY BACK IN 1988. THE ARTICLE WAS ACTUALLY PUBLISHED A YEAR AFTER GEORGE PASSED AWAY. HE PASSED AWAY IN APRIL, ON APRIL 25, 19 87. SO, AT THE TIME, GEORGE SAID TO ME, HE THOUGHT THAT THERE WOULD BE THREE WAYS MECHANISTICALLY, THAT WE COULD EXPLAIN HOW ENHANCERS WORK. SO OF COURSE WE ALL UNDERSTAND PROMOTORS ARE THE BASIC NITTY-GRITTY SEQUENCES ON TO WHICH THE RNA POLYMERASE CABBAGES UPON AND DRIVES TRANSCRIPTION. SO ENHANCERS IN GENERAL ARE UPSTREAM SEQUENCES THAT GEORGE THOUGHT WOULD IN FACT, SOMETHING WOULD ENHANCER WOULD COME HERE, AND IT WOULD TOP LOGICALLY CHANGE THE PROMOTOR MAKING THE PROMOTOR MORE ACCESSIBLE IN A WAY THAT WAS STILL SORT OF AT LEAST, BACK THEN IN ’88, NOT WELL UNDERSTOOD. HE ALSO THOUGHT THAT LOOPING WOULD BE IMPORTANT FOR ENHANCERS AND PROMOTORS TO SOMEHOW TOUCH EACH OTHER EACH THOUGH THIS WOULD BE IN THE 5 PRIME DIRECTION OR 3 PRIME DIRECTION AND THEREBY BRINGING — BRINGING FACTORS TO THE PROMOTOR DRIVING TRANSCRIPTION. AND IN ANOTHER WAY HE THOUGHT ENHANCERS WOULD WORK WOULD BE THAT THINGS WOULD BIND HERE AND THEN THEY WOULD SLIDE DOWNWARD AND THEY WOULD MEET THE PROMOTER AND DRIVE TRANSCRIPTION COMING FROM THE PROMOTER. SO I THINK RETROSPECTIVELY NOW, WITH THE BENEFIT OF THREE DECADES, WE KNOW THAT ALL OF THESE MODELS ARE IN ONE WAY OR ANOTHER APPLICABLE TO THE CONCEPT OF ENHANCERS. SO WHAT GEORGE ALWAYS EMPHASIZED TO ME WAS, AT THE END OF THE DAY, THE KEY FACTORS THATY WOO ARE GOING TO PAY ATTENTION TO, MUST BE THE BINDING PROTEINS. DNA BINDING PROTEINS BINDING TO ENHANCER AND BINDING TO PROMOTERS AND THAT IS IN FACT THE FINAL EFFECTORS THAT END UP DRIVING TRANSCRIPTION. SO, WHEN I LEFT HIS LAB TO START MY OWN LAB IN NIAID, THE VERY FIRST CHALLENGE THAT WE HAD WAS THINKING ABOUT THIS FACTOR THAT WAS A TRANSCRIPTIONAL TRANSACT VARIATE AND THIS IS THE HIV TAP PROTEIN THAT DRIVES TRANSCRIPTION FROM THE HIV LTR. AND AT THAT TIME, IT WAS VERY PUZZLING BECAUSE ALL THE LESSONS I LEARNED FROM GEORGE WAS THAT IT IS SUPPOSED TO BE A DNA BINDING PROTEIN. IT IS SUPPOSED TO DRIVE TRANSCRIPTION. AND IT HAS PHENOMENAL OBSERVATIONS THAT WOULD BE CONSISTENT WITH DNA-TYPE ENHANCER. BUT NONE OF THE EXPERIMENTAL EVIDENCE FITS WITH THAT. SO ONE OF THE THINGS I ALSO LEARNED FROM GEORGE IS THAT WHEN THINGS DON’T LOOK COMPLETELY RIGHT, YOU NEED TO PIVOT YOUR ANALYSIS. AND SO AT THE MOMENT WHEN WE TRIED TO EXPLAIN WHAT WAS GOING ON, I HAD AN EPIPHANY OR A NEW THOUGHT THAT PERHAPS IN FACT, IT IS NOT DNA BINDING, BUT IN FACT, ALTHOUGH AT THAT TIME, THERE WERE NO EXAMPLES OF SUCH A THING, AND IN FACT IT WAS A TRANSCRIPTIONAL ACTIVATOR THAT BOUND TO A MASON RNA DRIVEN FROM THE BASAL PROMOTER. SO IN FACT, THAT PARTICULAR NOTION TURNED OUT TO BE CORRECT AND WE PUBLISHED A PAPER IN CELL, WHICH DEMONSTRATED CAT AS A FIRST EXAMPLE OF AN RNA BINDING PROTEIN BINDS FROM THE HAIR PIN RNA AND THIS WAY WORKS LIKE A DNA ENHANCER BUT IS NOT A DNA ENHANCER BUT IS A DNA ENHANCER-LIKE PROPERTY THEY BINDS TO RNA AND REPRESENTED THE FIRST EXAMPLE OF TRANSCRIPTIONAL ACTIVATION THROUGH RNA BINDING LEADING TO ENHANCED TRANSCRIPTION FROM THE PROMOTER. SO THAT OPENED UP AN INTEREST IN KLINEING RNA BINDING PROTEINS AND WE ENDED UP CLONING ONE OF THE FIRST RNA BINDING PROTEINS IN HUMAN CELLS AND WE COINED THE NAME FOR THAT RNA BINDING PROTEIN, TRBP, FOR TAR RNA BINDING PROTEIN. SOME OF YOU YOUNGER PEOPLE IN THE AUDIENCE WILL HAVE ABSOLUTELY NO CLUE THAT THIS IS WHAT WAS THE ORIGIN OF THIS PROTEIN. WHEN I SAY TRBP TO YOU, YOU WOULD HAVE OF COURSE IMMEDIATELY SAID, OF COURSE TRBP. THAT IS IN FACT THE CRITICAL FACTOR WITH DICER FOR THE PROCESSING OF SMALL MICRORNAs. AND YOU WOULD OF COURSE BE RIGHT. BUT TRBP WAS ORIGINALLY IDENTIFIED AS A TAR RNA BINDING PROTEIN AND THE REASON IT WORKS AS A MICRORNA IN FACTOR IS BECAUSE IF YOU — IS BECAUSE IF YOU LOOK AT TAR RNA, NOW WITH THE BENEFIT HINDSIGHT 2020, IT LOOKS LIKE A SMALL MICRORNA. THOSE WERE THE TEACHINGS I APPRECIATE FROM GEORGE IN TERMS OF LOOKING AT TRANSCRIPTION AND LOOKING AT WAYS OF THINKING TO PIVOT YOUR NOTIONS IF IN FACT THE EVIDENCE SUGGESTS TO YOU THAT THE PHENOL NOLOGY IS SIMILAR BUT THE MECHANISM MIGHT BE DIVERSE. NOW THE REASON I WANT TO TALK ABOUT HTLV1 IS BECAUSE IT WAS A SHARED INTEREST BETWEEN JOHN BRADY, GEORGE KHOURY AND I AND IN FACT, THESE TWO PAPERS THAT WERE PUBLISHED BACK-TO-BACK IN CONSECUTIVE YEARS IN 19 KETCH AND 1988 AND INSTRUMENTAL AT THE TIME IN TERMS OF DEFINING HOW THE COUSIN OF THE HIV PROTEIN, THE HTLV1 KATZ PROTEIN WORKS IN TERMS OF MOTIF IT WAS RECOGNIZING AND IN TERMS OF A PATHWAY IN WHICH IT WAS USED FOR ACTIVATING TRANSCRIPTION. SO IT’S REMINISCENT OF THE ENHANCER OF THE SO THESE ARE DNA BINDING MOTIFS. AND THE SIGNALING PATHWAY THAT THE TAX PROTEIN ACTIVATES TRANSCRIPTION IS THROUGH THE CYCLIC AMP SIGNALING PATHWAY. SO, THESE ARE PAPERS ON WHICH JOHN AND I AND GENERAL AND OTHERS COLLABORATED ON IN THOSE EARLY DAYS, WHICH I THINK REALLY PAVEED THE INITIAL ROAD MAP IN TERMS OF OUR UNDERSTANDING OF THE BEGINNING OF THE HTLV1 LIFE CYCLE WHICH OF COURSE AS THE VIRUS ENTERS AND BECOMES A PROVIRUS IS AT THE LEVEL OF TRANSCRIPTION. NOW I KNOW MANY OF YOU ARE VERY FAMILIAR WITH HIV AND I KNOW PROBABLY MOST OF YOU ARE NOT FAMILIAR WITH HTLV1. SO I THINK ONE OF THE THINGS THEY WANTED DO IS BEFORE I TELL YOU THE SHORT STORIES I’M GOING TO TELL YOU ABOUT TH — HTLV1 IS TO GIVE YOU A BACKGROUND ABOUT THIS PARTICULAR VIRUS AND THE DECEASED IT CAUSES. SO, HUMAN T-CELL LEUKEMIA VIRUS TYPE I WAS THE FIRST HUMAN RETROVIRUS AND THAT WAS DISCOVERED BY BOB GALLOW. AND THE VIRUS IS THE CAUSATIVE AGENT OF A T-CELL LEUKEMIA CALLED ADULT T-CELL LEUKEMIA. THE INTERESTING NATURAL HISTORY OF THE VIRUS IS THAT THE VIRUS INFECT THE HUMAN AND 20-30 YEARS LATER, YOU HAVE WHITE BLOOD CELLS THAT END UP BECOMING LEUKEMIC CELLS. SO THERE IS A VERY LONG LATENCY PERIOD. SO SUGGESTING THAT THERE MUST BE MULTISTEP COMPLICATED PROCESSES AFTER VIRAL INFECTION BEFORE THAT INFECTED CD4 POSITIVE T-CELLS ENDS UP BECOMING LEUKEMIC CELL. THE VIRUS IS RESPONSIBLE FOR ALL THE IMMUNE DISEASES, HLV ASSOCIATED MILOPATHY, AND PARESIS AND EFFECTS 20 INDIVIDUALS WORLDWIDE. SO IT’S NOT BY ANY MEANS AN OVER AN DISEASE BECAUSE IN REALITY, THE NUMBER OF INDIVIDUALS INFECTED WITH HIV AT ANY GIVEN TIME IS AROUND 30 TO 35 MILLION INDIVIDUALS WORLDWIDE. SO, ESSENTIALLY THE NUMBER OF CAREIOTS FOR THLV1 IS ABOUT 60% OF THE NUMBER INDIVIDUALS INFECTED WITH HIV. NOW THE REASON WHY IT SEEMS TO BE VERY, VERY MUCH LESS VISIBLE IN THE UNITED STATES THAN ELSEWHERE IN PLACES LIKE JAPAN AND SOUTH AMERICA AND PARTS OF AFRICA, IS SIMPLY BECAUSE WHILE JAPAN HAS A SIGNIFICANT PREVALENCE, SO DOES IRAN AND PART OF AFRICA AND SOUTH AMERICA, THE PREVALENCE OF T RADIO – HTLV IS REALLY MINIMAL IN THE CONTINENTAL UNITED STATES. SO VERY FEW OF YOU WOULD HEAR OF ANYBODY INFECTED WITH HTLV OR HAVE ADULT T-CELL LEUKEMIA AS A DOWNSTREAM SEQUEL I OF HTLV1 INFECTION. THAT IS THE REASON WHY THE DISEASE IS NOT REALLY VERY VISIBLE IN WELL-KNOWN IN THE UNITED STATES. ALTHOUGH, I THINK IT IS A WONDERFUL MODEL IN TERMS OF OUR ATTEMPTS AT UNDERSTANDING AND INITIAL STIMULUS AND INSIGHTING EVENT AND HOW IT LEADS ULTIMATELY TO TRANSFORMATION. SO IF YOU ARE A PHYSICIAN, CHANCES ARE YOU WILL END UP SEEING ON A VERY RARE BASIS, AND THESE ARE SLIDES PROVIDED TO ME BY A BRAZILIAN PHYSICIAN WHO SEES AS I SHOWED YOU ON THAT MAP, A VERY HIGH CONCENTRATION OF TH — HTLV1 INFECTED INDIVIDUALS IN BRAZIL. AND PEOPLE TEND TO COME IN WITH THESE CUTANEOUS DERMAL MANIFESTATIONS BECAUSE ORIGINALLY, AS MANY OF YOU MAY REMEMBER, THE VIRUS THAT WAS ISOLATED BY BOB GALLOW, WAS ORIGINALLY DIAGNOSED AS A PERSON WHO HAD CUTANEOUS T-CELL LYMPHOMA. SO THE T-CELLS, ONCE THEY BECOME TRANSFORMED AS TOM WALLEDDER MAN CAN EXPLAIN BETTER THAN I CAN, ENDS UP MOVING INTO THE DERMAL SPACE AND PRESENTING WITH THESE RASH-LIKE LESIONS. NEVERTHELESS, I THINK THE BAD NEWS FOR MANY OF THESE INDIVIDUALS IS THAT THIS IS AN AGGRESSIVE T-CELL LEUKEMIA AND IT TENDS TO BE IN TRACKABLE AND UNTREATABLE. SO THE OUTCOME IS VERY POOR. THEREFORE GIVING ADDITIONAL IMPETUS TO WHY WE SHOULD DO MORE TO UNDERSTAND THE GENESIS OF THIS LEUKEMIA. NOW FOR THE PURPOSES OF THE MOLECULAR BIOLOGY OF T46789 — HTLV, THIS IS THE GENOME ORGANIZATION, LOOKS VERY MUCH LIKE A PROTOTYPIC ANIMAL RETROVIRUS WITH GAG, POL AND EN — ENV AND OF WHICH THE KEY FACTOR I SPENT SORT OF MY CAREER STUDYING IS THIS 40 KILL DALTON PROTEIN CALLED TAX. WHAT HAPPENS WHEN A CD4 POSITIVE T LYMPHOCYTE, NORMAL T LYMPHOCYTE BECOMES INFECTED WITH HTLV1, AS I MENTIONED TO YOU AFTER A REMARKABLY LENGTHY LATENCY PERIOD, THE VIRUS UNDERGOES TRANSFORMATION AND GIVES A VERY PATH PNEUMONIC PICTURE, IF YOU TAKE ANY SORT OF PATHOLOGY BOARD EXAM AND YOU SEE THIS KIND OF FLOWER CONFIGURATION SHAPE, NUCLEUS, THIS IS INEVITABLY GOING TO BE ATL AND NOT ANY OTHER KIND OF LEUKEMIA. SO, WHEN I APPROACHED THE QUESTION THEN ABOUT TWO DECADES AGO IN TERMS OF HOW AND WHY MECHANISTICALLY DOES THIS NICE ROUND SHAPED NUCLEUS BECOME DAMAGED AND DISTORTED NUCLEUS, I THOUGHT MYSELF, THAT THE PATHOLOGY AND THE HISTOLOGY IS TELLING US SOMETHING. IT’S TELLING US THAT THE DISEASE MUST HAVE SOMETHING TO DO WITH HOST SCALE DAMAGE TO THE CONSTITUENTS OF THE NUCLEUS, WHICH OF COURSE IS THE DNA. SO THERE MUST BE WHOLE CELL DNA DAMAGE OR THERE MUST BE WHOLE CELL CHROMOSOMAL CHANGES IN ORDER FOR SUCH DISTORTION FROM THIS NICE WELL-ROUNDED SHAPE TO THIS VERY, VERY CONTORTED FLOWER-LIKE CONFIGURATION OF A NUCLEUS. SO I THOUGHT, AMONG MANY HYPOTHESES THAT ONE COULD ARK RIFE AT IN TERMS OF INVESTIGATING HOW AND WHY YOU GO FROM NORMAL SEED TO A BARRON SEED, THAT ONE OF THE LIKELY AND PLAUSIBLE HYPOTHESES WOULD BE TO INVESTIGATE THE QUESTION OF HOW YOU GO FROM VIRUS INFECTING THE CELL, GENERATING DISTORTED NUCLEAR MORPHOLOGY, WHICH PROBABLY SHOULD BE ACCOMPANIED BOY GROWS ANEUPLOIDY, MARKEDLY GROSS ANEUPLOIDY. AND I THINK GENERALLY THAT WOULD NOT BE A VERY BAD GUESS BECAUSE IN FACT, IF YOU LOOK AT WHOLESALE SERIES OF CANCERS, 70-80% OF THEM ARE LINKED TO THE ANEUPLOIDY OF THEIR GENOMES. BY COMPARISON, THE NUMERICAL LINKAGE IS p53 MUTATIONS AND SOMEWHAT NORTH OF 50%. AND OF COURSE, ONE OF THE STRIKING CORRELATIONS IS THAT IF YOU LOOK AT ATL CELLS, ALL OF THEM ARE ANEUPLOIDY. SO 100% OF THEM. SO SUGGESTING THAT, AS I THINK MANY ONCOLOGISTS AND CANCER BIOLOGISTS COMING AROUND, WHICH IS ANEUPLOIDY, COULD POTENTIALLY BE THE CAUSAL REASON FOR ONCOGENESIS AS OPPOSED TO THE ARGUMENT, WHICH HAS BEEN FLOATING AROUND FOR MANY YEARS, WHICH IS IN FACT, ANPLOYED SEPERHAPS THE CONSEQUENCE OF TRANSFORMATION AND NOT NECESSARILY THE CAUSE OF TRANSFORMATION. SO, IF ONE WERE THEN TO ACCEPT THE HYPOTHESES THAT ANEUPLOIDY IS A WORTHWHILE NOTION TO EXPLORE FOR HTLV GENESIS OF ATL, THEN HOW DOES ONE GO ABOUT MECHANISTICALLY TRYING TO UNDERSTAND HOW THAT ARISES? IF YOU THINK ABOUT THE GENESIS ANEUPLOIDY, BASICALLY SOMETHING BAD HAS TO HAPPEN IN MITOSIS, BECAUSE DURING MITOSIS IS WHEN THE 46 CHROMOSOMES GET DUPLICATED AND WHEN THEY ARE SUPPOSED TO SEPARATE WITH NUMERICAL FIDELITY SO THAT WHEN THE 46 CHROMOSOMES ALL BECOME DOUBLE, THE SPINDLE POLL CATCHES THEM AND HAS EQUAL SEGREGATION SO THE ONE MOTHER CELL THAT WEEKS MITOSIS, ENDS UP WITH TWO DAUGHTER CELLS, BOTH OF WHICH MAINTAINS UPLOYEDY. THOSE UPLOYEDIES OCCUR BECAUSE THERE ARE TWO POLLS. YOU CAN SAY THIS IS THE WEST POLL AND THIS IS THE EAST POLL AND THEY THROW UP ROPES, TIED TO THE KINETIC COURSE OF THE CHROMOSOMES AND THEN THERE IS EQUAL TENSION AND EQUAL PULLING SO THEN YOU GET EQUAL FIDELITY OF SEGREGATION. BUT WHAT HAPPENS, AND THIS HAPPENS QUITE FREQUENTLY IN CANCER CELLS, WHICH IS THAT THE BODIES HAVE BECOME SPINDLE POLES, SOMETIMES MISS DUPLICATE. SO INSTEAD OF TWO SPINDLE POLES, YOU COULD HAVE 3 SPINDLE POLES OR YOU COULD HAVE 4 SPINDLE POLES AND IN THAT CASE, THEN, IT WOULD BE AN UNEQUAL TUG-OF-WAR BECAUSE WHEN EXTRA ROPES ARE THROWN FROM THESE THREE SPINDLE POLES AND THEY HAVE TO BE MATCHED AGAINST THE OTHER ONE SPINDLE POLE, YOU CAN IMAGINE HOW THREE POLES PULLING WOULD PULL MORE CHROMOSOMES INTO ONE CELL THAN ONE POLE PULLING IN THE OPPOSITE DIRECTION. SO THEREBY YOU HAVE THE GENESIS AND THE OUTCOME OF SIMPLE PHYSICS WHERE MORE FORCE GOES IN THIS WAY, LESS FORCE GOING THIS WAY AND MORE FORCE GOING THIS WAY AND SO MORE CHROMOSOMES GOING THROUGH ONE DAUGHTER AND WE’LL FEWER CHROMOSOMES GOING WITH THE OTHER DAUGHTER CELL AND THEREFORE THE GENERATION OF ANEUPLOIDY CELLS. AND WE THINK THEN THAT THE GENERATION OF ANEUPLOIDY CELLS IS THEN ONE OF THE EARLY MANIFESTATIONS OF ATL AND THE ACCUMULATION OF A TIME IS HOW IT BECOMES A FRANK LEUKEMIC CELL DOWN THE ROAD. SO IF THAT IS THE CASE, AND IF WE WERE TO UNDERSTAND THE ATTACKS PROTEIN IS THE ONCOGENIC FACTOR FOR ATL, THEN HOW DOES TAX GO AROUND CAUSING THESE ABARENT NUMBERS OF NUMERICAL SEPARATION? THE ANSWER TURNS OUT TO BE ONE OF THE FINDINGS WE CAME UP WITH, IS IF YOU MAKE A FLORESCENT TAX PROTEIN, WHAT YOU FIND IS THAT WHEN YOU INTRODUCE IT INTO CELLS, EVERY TIME YOU SEE THE CONSTITUENT PROTEIN OF THE SPINDLE POLE, OR THE CENTROSOME, EVERY TIME YOU STAIN FOR PER I SEN TRIN, YOU FIND THAT THE GREEN TAX PROTEIN ALWAYS GOES WHEREVER THE POLE IS SUPPOSED TO BE SOLD FUELED FIND TWO BRIGHTLY GROWN TAX STAINED POLAR BODIES. IF YOU FIND THREE, THIS IS AN ABARENT MITOSIS BECAUSE THERE IS A TRY POLER AS OPPOSED TO BY POLER. AND THIS KIND WILL ALWAYS LEAD TO AN ANEUPLOIDY CELL. THEN YOU FIND WHAT HAPPENS IS THERE ARE THREE GREEN SPOTS. SO SUGGESTING THAT EVERYWHERE WHEN YOU SEE A SPINDLE POLE BODY, THAT IS THE ACTUAL ANCHORS RESPONSIBLE FOR FIDELITY OF SEPARATION, YOU WILL FIND THE TAX PROTEIN HERE. AND IN FACT, WHAT WE THINK IS GOING ON IS THAT THE TAX PROTEIN, ACTUALLY ASSOCIATES WITH PARASEN TRIM PROTEINS EARLY AND CAUSES THOSE PARASEN TRIM PROTEINS BY COALESCING ON THEM, TO MISFRAGMENT SO THAT AT AN INCREASED RATE, THEY TEND TO FRAGMENT, INSTEAD OF INTO TWO PARTS, INTO MULTIPLE FRAGMENTS. AND THEREFORE, IN TAX EXPRESSING CELLS YOU HAVE A MUCH GREATER PROPENSITY FOR MULTIPOLAR MITOSIS AND THAT IN ITSELF BECOMES A PHYSICAL EXPLANATION FOR WHY YOU WILL END UP WITH THE INSIGHTING DRIVING FORCES FOR THE CREATION OF ANEUPLOIDY. THAT CAN’T BE THE WHOLE ANSWER. BECAUSE WHENEVER THIS TRIES TO GO ON OR IF YOU HAVE A MISS ATTACHMENT OF SPINDLES, THE MICROTUBULAR FROM THE SPINDLES, WHEN THESE EVENTS ARE SUPPOSED TO BE GOING ON, WE KNOW THE NORMAL CELLS HAVE A CHECKPOINT THAT SAYS, YOU MAY FORM THESE THINGS, BUT THE CHECKPOINT OVERRIDES YOU AND SAY YOU MAY NOT EXIT MITOSIS AND YOU MAY NOT DIVIDE. SO WHAT IS THAT CHECKPOINT? THAT CHECKPOINT OF COURSE IS THE MITOTIC SPINDLE ASSEMBLY CHECKPOINT. SO, THERE ARE SEVERAL CHECKPOINTS IN THE CELL CYCLE. BUT I TEND TO THINK ABOUT THEM AS TWO MAJOR CHECKPOINTS. THERE IS ONE MAJOR CHECKPOINT BY ONE OF GEORGE’S FAVORITE PROTEINS BECAUSE ACTIVATED BY SB40 LARGE ANDROGEN AND p53, THAT IS A CHECKPOINT FOR STRUCTURAL DNA DAMAGE AS THE DNA GROWS G1 TO S TO BECOME REPLICATED TO BE COPIED. SO STRUCTURAL DAMAGES MUST BE CLEARED UP BEFORE p53 WILL ALLOW TRANSITION FROM G1 TO S. HERE NOW, THE STRUCTURAL GAME IS OVER. WHAT IS IMPORTANT HERE IS THE NUMERICAL GAME. BECAUSE WHEN YOU EXIT MITOSIS, YOU ARE SUPPOSED TO HAVE FIDELITY OF NUMERICAL SEPARATION, THE STRUCTURAL MONITORING IS DONE HERE AND NUMERICAL MONITORING IS DONE HERE. AND IF, IN FACT, EVEN IF YOU HAD THESE UNEQUAL TUG-OF-WARS, THEY ARE NOT SUPPOSED TO EXIT MITOSIS, AS LONG AS THE SPINDLE ASSEMBLY CHECKPOINT IS INTACT. SO WE KNEW THAT IN TAX EXPRESSING CELLS IN ATLs, MULTIMITOSIS OCCURS AND MULTIPOLAR MITOSIS ARE ALLOWED TO IN FACT CONSUMMATE TO PRODUCE ANEUPLOIDY CELLS. SO SUGGESTING TO US THEN SOMETHING ELSE MUST BE HAPPENING TO THIS CHECKPOINT, EVEN THOUGH WE HAD ONLY IDENTIFIED THE TAX PROTEIN AS ASSOCIATING WITH THE SPINDLE POLE BODIES. SO, IN ORDER TO ANSWER THIS QUESTION, THEN, A POSTDOCTORAL FELLOW WHO JOINED THE LAB DECIDED THAT THIS MUST BE A NUCLEAR INTERACTION OCCURRING DURING MITOSIS. SO HE WENT AHEAD AND HE CLONED EVERY SINGLE TAX BINDING PROTEIN IN THE MITOTIC PHASE OF THE CELL CYCLE THAT BOUND THE TAX PROTEIN. AND FROM THAT CLONING, WE IDENTIFIED THE PROTEIN THAT WAS AT THAT TIME POSTULATED TO EXIST FROM EAST MITOTIC EXPERIMENTS AS IN MY TO THETIC DEFICIENCY PROTEIN NUMBER 1. SO THIS TURNS OUT TO BE THE MITOTIC SPINDLE ASSEMBLY CHECKPOINT PROTEIN THAT THE TAX PROTEIN, ALSO HAS TO BIND TO AND ALSO HAD TO END UP ENACTEDIVATING. SO IT TURNS OUT THERE ARE TWO DIFFERENT EVENTS GOING ON. ONE IS THE EVENT THAT CAUSES THE PHYSICAL REASON FOR GENERATION OF ANEUPLOIDY. AND THEN THERE IS THE SECOND EVENT WHICH IS THAT THE TAX PROTEIN HAS TO IN ACTIVATE THE CENTURY, THE POLICE MAN THAT SAYS, WHEN SOMETHING BAD HAPPENS, YOU ARE STILL ALLOWED TO LIVE AND YOU MUST NOT BE SENT INTO APOPTOSIS AND BE COMMITTED TO THAT. SO BY DOING TWO DIFFERENT THINGS, CREATING THE PHYSICAL REASON FOR ANEUPLOIDY AND ALSO INACTIVATING THE CHECKPOINT THAT SENSORS AGAINST ANEUPLOIDY, WE BELIEVE THAT A TAX PROTEIN ALLOWS THE CONSUMMATION OF THE FORMATION OF ANEUPLOIDY CELLS, WHICH ARE IN FACT THE LEADING EVENTS, EARLY EVENTS FOR THE GENESIS OF ATL LEUKEMIA. WHEN WOO PUBLISHED THIS PAPER IN “CELL,” THERE WAS A LOT OF BROUHAHA. PEOPLE SAID THIS PAPER REALLY DOESN’T ANSWER THE QUESTION OF SPINDLE ASSEMBLY CHECKPOINT AND CANCER. ALL IT DID REALLY SAY WAS, THE MAD ONE PROTEIN BINDS TO AN OVER EXPRESSED ONCOPROTEIN. AND WE REALLY DON’T KNOW WHETHER THE ONCOPROTEIN IS SIMPLY ONLY INACTIVATING THE MAD ONE PROTEIN TO CAUSE TRANSFORMATION AND DEFECTS OR WHEN YOU OVER EXPRESS THE ONCOPROTEIN, THERE ARE A LOT OF THINGS YOU DON’T CHOOSE TO SEE, THEY ARE ACTUALLY RESPONSIBLE FOR THE CHANGES — NOT COMPLETE IN TERMS OF DEMONSTRATING THIS CHECKPOINT AND THIS PROTEIN HAS ANY ROLL IN TERMS OF TRANSFORMATION. SO WE SORT OF FRETTED OVER THAT FOR A WHILE AND THEN WE FINALLY DECIDED THAT THE SIMPLEST WAY TO ANSWER THAT QUESTION WAS SIMPLY TO MAKE THE KNOCKOUT. SO IF YOU KNOCKOUT THIS PARTICULAR CHECKPOINT PROTEIN AS MANY OTHER CHECKPOINT PROTEINS HAVE BEEN DONE WITH, IF THE CHECKPOINT PROTEIN IS IMPORTANT FOR GUARDING AGAINST TUMORIGENESIS, THEN WE MOVING THE CHECK POINT PROTEIN SHOULD BE IN A VERY SIMPLE WAY SHOW UP INCREASING TUMORIGENESIS IN THE KNOCKOUT ANIMAL AND THAT EXPERIMENT TOOK US A FEW YEARS TO DO BECAUSE WE WERE JUST LEARNING HOW TO DO KNOCKOUT EXPERIMENTS AND WE ENDED UP CREATING HET REZYGOUS KNOCKOUTS. AND THE HOMOZYGOUS KNOCKOUTS TURNED OUT TO BE EM BRIE ONICALLY LETHAL. IF IT IS IMPORTANT AND YOU LOSE ONLY 50% OF THE FUNCTION OF THAT CHECKPOINT, IT SHOULD STILL BE SUFFICIENTLY CLEAR TO MANIFEST INCREASE IN THE NUMBER OF TUMORS. AND IN FACT, IF DID TURN OUT TO BE THE CASE. SO IN A FAIRLY LARGE SERIES OF MICE, SO OVER 100 MICE, WILDTYPE MICE, WE ALL KNOW THAT WILDTYPE MICE HAS A BASAL LEVEL OF TUMORIGENESIS SPONTANEOUS TUMORIGENESIS UPON AGING. WHEN YOU AGE THESE MICE, BY THE TIME YOU REACH TWO YEARS, SPONTANEOUS TUMORIGENESIS IS ABOUT 18 OUT OF 130 AND THE HETEROZYGOUS KNOCKOUT OF THIS CHECKPOINT PROTEIN SHOWS UP DOUBLING THE RATE WITH VERY SIGNIFICANT P VALUES OF .0043 SO SUGGESTING THAT OUR PARTICULAR HYPOTHESES IS CORRECT. INACTIVATION OF THIS PARTICULAR CHECKPOINT LEADS TO TUMOR — INCREASED ANPLOYEDY AND INACTIVATION BY THE AN COPROTEIN CREATES THAT GENERATES THAT AND ULTIMATELY CAUSES TUMORIGENESIS. NOW WE CAN EMBELLISH UPON THAT EXPERIMENT IN THE FOLLOWING FASHION. WE KNOW NOW IN THE EARLY DAYS, WE KNEW LESS, BUT WE KNOW NOW THAT IN FACT MAD ONE IS ONE OF SEVERAL COMPONENTS OF THIS CHECK POINTED. MANY OF THESE ARE REDUNDANT, ALTHOUGH MANY ARE ALSO INDEPENDENTLY CRITICAL. SO WE KNOW IN FACT THE MAD 1 AND MAD 2 PROTEINS ARE HETERODIMERIC PARTNERS. SO WE OF COURSE MADE A WEAK NONFUNCTIONAL CHECKPOINT BECAUSE WE COULD ONLY MAKE THE HETEROZYGOUS ANIMAL. WE THOUGHT TO OURSELVES THAT PERHAPS THE EXPERIMENT COULD BE REINFORCED THAT THE NOTION AND HYPOTHESES COULD BE REINFORCED. IF WE COULD MAKE COMPOUND ANIMALS WITH THE KNOCKOUT OF ALSO THE MAP TWO PROTEIN AND MAD 1 PROTEIN AND MARRY THESE MICE TOGETHER TO MAKE BABIES THAT WERE IN FACT DOUBLING UP FOR MAD 1 AND MAD 2. NOW EACH HOMOZYGOUS KNOCKOUT IS LETHAL SO WE COULD MAKE COMPOUND HETEROZYGOUS KNOCKOUTS. WE GENERATED A NUMBER OF INDEPENDENT MOUSE EMBRYO FIBROBLASTS CLONES THAT ARE WILDTYPE MICE THAT ARE HETEROZYGOUSLY KNOCKED OUT FOR ONE CHECKPOINT PROTEIN. HETEROZYGOUS KNOCKOUT FOR THE SECOND AND THOSE THAT ARE DOUBLY HETEROZYGOUS KNOCKED OUT FOR BOTH OF THE CHECK POINT PROTEINS. AND SO THE PREDICTION AND CELL BIOLOGY VERIFIES A PREDICTION THAT THERE IS AN INCREASED PROPENSITY TOWARDS AN PELOSI AS YOU GO FROM WILDTYPE TO THESE KNOCKOUTS. MAD TWO IS MORE CRITICAL THAN MAD ONE. MAD ONE TENDS TO BE A REGULAR PARTNER OF MAD TWO. AM AND THE BOTTOM LINE IS THAT AS YOU GO INCREASING IN THE DEGREE OF PROPENSITY TOWARDS AN LIKEWISE I YOU DECREASE TUMORIGENESIS. MAD 1 ARE MORE TUMOR PRONE AND MAD TWO ARE EACH MORE. AND THIS EXPERIMENT WE SHOW YOU SOME EXAMPLES HERE AND BUT IT HAS BEEN REPLICATED AND INDICATING THE WHOLE IDEA IS TRUE. THAT THE MORE YOU GENERATE IN TERMS OF CELLS, THAT TEND TO BECOME ANEUPLOIDY, THE MORE POTENT THEY ARE IN TERMS OF TUMORIGENESIS WHEN INTRODUCED BACK INTO THESE ANIMALS. SO THEN YOU SAY TO YOURSELF, BUT AT THE END OF THE DAY, THESE ARE STILL MICE. RIGHT? WHAT IS THE EVIDENCE FOR MAD ONE INACTIVATION HUMAN CANCERS? AND YOU LOOK VERY HARD AND YOU LOOK FOR GENETIC MUTATIONS OF MAD 1. YOU DON’T FIND THEM VERY FREQUENTLY. SO A LOT OF PEOPLE ARGUED THESE ARE INTERESTING AND FINE OBSERVATIONS IN MOUSE SYSTEMS BUT PERHAPS NO RELEVANCE TO HUMAN SYSTEM SYSTEM. IT TURNS OUT THAT’S NOT CORRECT BECAUSE IN FACT YEARS AGO WHEN WE SURVEYED COLON CANCERS, WE FOUND THAT THE GENETIC EVIDENCE IS CORRECT. INACTIVATION OF MAD 1, GENETICALLY IN TERMS OF MUTATION, IS IN FACT VERY RARE. WHAT WE FOUND WAS OVER 80% OF THE COLON CANCERS HAVE LOST MITOTIC SPINDLE ASSEMBLY CHECKPOINT FUNCTION BY OVEREXPRESSING THE MAD ONE PROTEIN. SO AT THE TIME WHEN WE REPORTED THAT, PEOPLE SAID, NO, I MEAN, THAT’S JUST SORT OF THEIR RATIONALIZATION BECAUSE THEY CAN’T BIND MAD 1, GENETIC MUTATIONS. SO IT TURNS OUT THAT A COUPLE OF MONTHS AGO, AS I WAS READING THROUGH CATCHING UP ON THE MEAD ONE LITERATURE, THIS PARTICULAR PAPER WAS PUBLISHED IN THE PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCE BASICALLY AND ALWAYS GRATIFYING WHEN PEOPLE IN THEIR ABSTRACT SITE YOUR PAPER OUTRIGHT AT THE BEGINNING AND SAY, BASED ON WHAT THEY FOUND, FIVE YEARS AGO, WE WENT AND LOOKED AND IN THAT CASE THEY LOOKED IN HUMAN BREAST CANCERS AND THEY FOUND THAT OVER 70% OF HUMAN BREAST CANCERS BEHAVE LIKE WHAT WE HAD SEEN WITH COLON CANCERS, ALL OF THEM, ALL 70% OF THOSE ARE INACTIVATED FROM MITOTIC SPINDLE ASSEMBLY CHECKPOINT BECAUSE BY VIRTUE OF OVEREXPRESSING THE MAD ONE PROTEIN AT THE PROTEIN LEVEL AS OPPOSED TO GENETIC MUTATION OF THE MEAD ONE GENE IN TERMS OF INACTIVATING THAT PARTICULAR PROTEIN. SO OVER EXPRESSION AS WELL AS LOSS OF EXPRESSION, BOTH END UP INACTIVATING THE SPINDLE CHECKPOINT AND THE KEY MESSAGE IS INACTIVATION OF THE SPINDLE ASSEMBLY CHECKPOINT WHETHER BY GENETIC MUTATION OR OVEREXPRESSION, IN THE MOUSE CASE, WE CAN DEMONSTRATE BY GENETIC MUTATION. IN THE HUMAN CASE IT IS OCCURRING BY OVEREXPRESSION OF THE MAD 1 PROTEIN AND LEADS TO THE SAME FINAL OUTCOME WHICH IS CANCER ONCOGENESIS. NOW, IN THE FIRST PART I TALKED ABOUT HTLV1 AND ANEUPLOIDY AND I WANT TO SWITCH GEARS A LITTLE BIT AND TALK ABOUT A SHORTER STORY, WHICH IS ABOUT HTLV1 AND SANE DIP TEE. ONE OF THE FAVORITE QUESTIONS I’M ALWAYS ASKED BY MY COLLEAGUES OUTSIDE OF NIH, WHAT IS THE BENEFIT OF YOU GUYS WORKING AT THE NIH? OTHER THAN THE FACT THAT YOU HAVE TO FILL IN ALL SORTS OF ETHICS FORMS AND YOU DON’T GET APPROVED TO TRAVEL WITHOUT ETHICS SORT OF GOING THROUGH THE FINE-TOOTH COMB? I ALWAYS SAY ONE OF THE BENEFITS HAVE COMMUNITY RESEARCHERS OF NIH, IS THAT WE GET TO BE ABLE TO FOLLOW SERENDIPITOUS FINDINGS AND A LOT OF TIMES, THAT ENDS UP BEING SOME OF OUR BEST SCIENCE. SO, PART OF OUR SERENDIPITOUS FINDING THAT I’M GOING TO TELL YOU, THIS IS A MUCH MORE RECENT STORY, WHICH IS THAT — AND IT FOLLOWS FROM OUR STUDY OF THE MAD ONE PROTEIN. WE WANTED TO UNDERSTAND HOW THIS PARTICULAR PROTEIN ENDS UP FROM OF COURSE TRANSALATION INTO THE CYTOPLASM AND AT SOME POINTED, HAVING TO GO INTO THE NUCLEUS AND GO INTO THE SPINDLE POLE BODIES AND SITTING ON KINETIC COURSE, HOW DOES IT GO? WHAT IS THE JOURNEY OF THIS PARTICULAR PROTEIN? AND COULD WE LEARN SOMETHING FROM WHERE IT STOPS AND WHAT PARTNERS IT HOLDS HANDS WITH? SO, BY-AND-BY, AS WE WERE STUDYING THE TRANSLATED PROTEIN AND AS IT MOVES INTO THE NUCLEUS, POISED WHEN MITOSIS BEGINS IT TO GO TO THE KINETIC CORES, AND SPINDLE POLE BODIES, WE FOUND AT THAT TIME A CRUDE PROTEOMICS ANALYSIS THAT THE PROTEIN WAS IN FACT BINDING TO AN INNER NUCLEUS PROTEIN CALLED SUNG ONE. SO I SAID TO THE POSTDOC, I SAID, WELL, YOU KNOW, HERE IS A SIMPLE EXPERIMENT TO DO BECAUSE NOW WE HAVE GOTTEN MUCH BETTER AT MAKING KNOCK KNOCKOUT MICE. WE SHOULD — THIS IS A REGULATORY PROTEIN OF THE MAD 1 PROTEIN. MAD 1 HAS TO SIT HERE AND BOUND TO THIS INNER NUCLEO PROTEIN BEFORE IT GOES TO DOING MITOTIC SPINDLE ASSEMBLY CHECKPOINT FUNCTION. WE KNOCK THIS GUY OUT, THE WHOLE SPINDLE ASSEMBLY CHECKPOINT FALLS APART. MAD 1 KNOCKOUT IS HOMOZYGOUSLY LETHAL. MAYBE THIS IS NOT SO IT GIVES US A CHANCE TO ANSWER THIS QUESTION IN A CLEAN FASHION. SO POSTDOC AGREE AND WE MADE A KNOCKOUT OF THIS PARTICULAR PROTEIN AND LOW AND BEHOLD WE WERE EXCITED BAUDS THIS ONE WE COULD KNOCKOUT AND WE COULD BREED TO HOMOZYGOSITY. SO WE SAID, OKAY. NOW, THE WAY LAY STATION FOR MAD 1 ON ITS WAY TO KINETIC CORE IS NOW DESTROYED. SO THE CHECKPOINT MUST BECOME INACTIVATED. ALL WE HAVE TO DO IS SIT HERE AND LOOK AT THESE MICE AND THEY ARE BORN ALIVE AND SO WE CAN FOLLOW THEIR PHENOTYPE AND LET’S JUST WAIT FOR TUMORS TO FORM. AND WE WAITED AND WAITED AND WAITED AND EVERYTHING WAS NORMAL. NO TUMORS. AND SO, THE POSTDOC WORKED VERY, VERY HARD BECAUSE YOU SPEND A LOT OF TIME DEVELOPING AN KNOCKOUT MOUSE AND YOU REALLY NEED TO GET INSIGHT INTO IT. SO SHE FOUND THAT IN FACT, NOTHING WAS VERY REMARKABLE EXCEPT THAT THESE ANIMALS HAVE PROBLEMS WITH MEALIO GENESIS. SO IN THIS CASE, SPERMS DON’T FORM VERY WELL. AND OF COURSE, THESE SMALL RNAs, PRIOR RNAs DIDN’T FORM. WE PUBLISHED A PAPER DESCRIBING FINDINGS BUT IT WAS UNSATISFYING BECAUSE OUR ORIGINAL MODEL WAS THAT THIS WAS SUPPOSED TO LEAD TO A DISEASE MANIFESTATION AND WE WERE REALLY INTERESTED IN FINDING A DISEASE PHENOTYPE. SO AROUND THIS TIME, I WAS FRETTING ABOUT THIS AND I SAID, THIS IS AN INNERNIQUEULAR ENVELOPE PROTEIN. MAYBE I SHOULD READ UP ABOUT NUCLEAR ENVELOPE PROTEINS WHEN THEY ARE NOT WORKING, WHAT SORT OF DISEASE THEY ARE SUPPOSED TO CAUSE, BECAUSE AGAIN IT’S A NEW AREA FOR ME. SO I CAME ACROSS THIS REVIEW PAPER AT THAT TIME WHICH WAS PUBLISHED BY COLIN STEWART AND COLIN USED TO BE AT NCI FREDRICK BEFORE MOVING TO SINGAPORE. SO COLIN IS OF COURSE AN EXPERT IN NUKELY ENVELOPE PROTEINS AND HE WROTE THIS VERY, VERY ILLUMINATING ARTICLE BASICALLY DESCRIBING THAT A NUMBER OF HUMAN GENES AND PROTEINS THAT ARE RESPONSIBLE WHEN THEY HAVE BEEN LOSS FOR FUNCTION, THEY ARE RESPONSIBLE FOR NUCLEAR ENVELOPE DISTORTIONS AND ABERRATIONS AND THOSE TURN OUT TO BE MUSCULAR DISTROPEIC TYPE OF DISEASES. OF WHICH, ONE OF THE MORE IMPORTANT CATEGORY, ARE THE LAMINATING MUTATION AND THE — RESPONSIBLE FOR DISEASE JUST LIKE MUSCULODYSTROPHY AND LAM INA IS RESPONSIBLE FOR THIS FAMOUS DISEASE OF WHICH FRANCIS COLLINS IS THE WORLD RENOWNED EXPERT ON. SO I SAID, I DON’T UNDERSTAND. EVERYBODY ELSE’S NUCLEAR ENVELOPE GENE KNOCKOUT ENDS UP DEVELOPING INTO THESE VERY, VERY TRAUMATIC PHENOTYPES AND OUR KNOCKOUT AND THIS PROTEIN IS ONE OF THE MAJOR COMPONENTS OF THE NUCLEAR ENVELOPE. AND OURS JUST SORT OF MICE JUST LOOKED BACK AND BLINK AT YOU AND THERE IS REALLY NO PHENOTYPE EXCEPT THE MICE DON’T LIKE TO REPRODUCE CHILDREN AND THEY ARE INFERTILE. SO I SAID, WHAT CAN WE DO ABOUT THIS? AND SO SITTING DOWN DISCUSSING WITH A POSTDOC, I THOUGHT, PERHAPS WHAT THIS IS, IS THAT THIS IS NOT A MAJOR FACTOR BUT MAYBE IT’S A CO-FACTOR. SO WE KNOW THAT THERE ARE MOUSE MODELS FROM COLLINS STEWART, KNOCKOUT FOR LAMIN A AND THOSE TENDS TEND TO REPLICATE WHAT PEOPLE SEE IN HUNTING TON PROJARE YOU’RE YASSIN CHROME, THEY HAVE A RAPID AGING PHENOTYPE AND THEY DIE PREMATURELY. SYNDROME — THIS MOUSE DIES WITHIN 50 DAYS AFTER BIRTH. THAT IS EXTREMELY PREMATURE AGING. SO IF WE THINK THAT THIS GUY DOES NUCLEAR DAMAGE AND NUCLEAR ENVELOPE DISORDERS, IF WE MARRY THIS NOWS OUR SUN 1 KNOCKOUT MOUSE, WOULD WE NOT END UP WITH A MORE DRAMATIC NUCLEAR DISTORTION AND END UP DEVELOPING A FASTER AGING MODEL AND IN FACT THAT WOULD BE THE CATS MEOW AND PEOPLE INTERESTED IN AGING WOULD WANT TO AND AND ASKS FUR OUR MOUSE MODEL. I SUGGESTED THAT TO THE POSTDOCTORAL FELLOW AND SHE AGAIN WAS VERY ACQUIESCE ENT ABOUT IT AND CREATED THE COMPOUND MARRIED MOUSE IN WHICH WE THOUGHT WOULD END UP INTO AN EACH MORE LETHAL FASTER AGING PHENOTYPE. AND LOW AND BEHOLD WHAT WE FOUND WAS, AS YOU RECALL, I TOLD YOU, THIS PARTICULAR SINGLE KNOCKOUT RESULTS IN 100% LETHALITY BY DAY 50. ALTHOUGH ALL THE MICE ARE BORN NORMAL. INTERESTINGLY, AND THIS IS SOMETHING THAT TELLS ME THAT MY MOTHER AND YOUR MOTHER PROBABLY TOLD YOU THE SAME THING, ISN’T ALWAYS RIGHT. BECAUSE IF YOU WERE, MOM USED TO ALWAYS SAY, YOU KNOW, TEH, TWO WRONGS DON’T MAKE A RIGHT. SO HERE IS ONE WRONG, NOW HERE WE CREATE TWO WRONGS AND NOW IN FACT WE RESCUE INSTEAD OF GREATER LETHALITY, WE END UP RESCUING THE MICE NOW BACK TO NORMAL AGING. ALMOST NORMAL AGING. THIS IS NORMAL AGING. THIS IS NORMAL AGING. BUT THIS IS PRETTY DARN GOOD BECAUSE THE P VALUE IS DOWN TO .0001. SO THIS TELLS THAT YOU SOMETHING IS GOING ON. AND THIS TELLS YOU THAT IN THIS MOUSE, LIES A CLUE TO WHY THIS MOUSE AGES PREMATURELY BECAUSE THIS MOUSE ENDS UP CURING THE AGING PHENOTYPE THAT IS COMING OUT OF THIS MOUSE. SO WHAT IS THE ANSWER? SO, WE END UP RECENTLY PUBLISHING THIS PAPER IN CELL. AND THE ANSWER TURNS OUT THAT THE INTERESTING OUTCOME OF ONE KNOCKOUT, WHICH IS A KNOCKOUT OF LAMIN A, IS THAT THE LAMIN A KNOCKOUT ITSELF, WE THINK, IS NOT DIRECTLY PATHOGENIC. WHAT HAPPENS IS THAT WHEN YOU LOSE THE LAMIN A PROTEIN, THE NUCLEAR ENVELOPE IS NOT SINT SIDES PROPERLY AND THE CELL SEEMS TO THINK THAT THE REASON IT’S NOT SYNTHESIZING PROPERLY IS BECAUSE IT’S NOT MAKING ENOUGH SUN 1 PROTEIN. SO IT KEEPS ON TURNING OUT THE SUN 1 PROTEIN AS COMPENSATION FOR THE LOSS OF LAMIN A SO WHAT YOU FIND IN LAMIN AMEFs IS YOU HAVE GOBS AND GOBS OF OVER ACCUMULATION OF SUN 1. AND OF COURSE, IT MAKES SENSE WHEN YOU KNOCKOUT THE SUN 1, THE LAMIN A LOSS IN ITSELF IS NOT PATHOGENIC. WHEN YOU KNOCKOUT SUN 1, YOU CAN’T OVERTURN THE SUN 1 PROTEIN. SO THEREFORE THE DIRECT REASON FOR PATHOGENESIS, WHICH IS OVEREXPRESSION AND THE AMOUNTY TO RID OF THIS OVERACCUMULATEDPLOYEDY, IS NOW SOLVED AND THEREFORE AGING COMES BACK TO NORMAL. SO IS THIS THE SAME WITH THE HUMAN EXAMPLE OF LAMIN A MUTATION, WHICH IS THE PRO JARE YASSIN DREAM? IT TURNS OUT THAT AT LEAST FROM WHAT WE HAVE SEEN TO ALSO BE THE CASE, BECAUSE IN HTPS, THERE ARE ALSO CELLS WHICH OVEREXPRESS SUN 1. AND ONLY THE CELLS THAT OVER EXPRESS SUN 1, SO THESE ARE NORMAL CELLS, THEY STAIN FOR SUN 1 AND IN HTPS, 1-THREE CELLS IS OVER EXPRESSED SUN 1 BUT ONLY THESE OVER EXPRESSED IN HTPS ARE DISTORTED IN TERMS OF NUCLEAR MORPHOLOGY AND ARE PATHOLOGICAL. THE ONES THAT ARE NOT OVEREXPRESSED FOR SUN 1, EVEN THOUGH EVERY ONE OF THEM HAVE LAMIN A MUTATION, SOME DON’T TURN OUT TO BE OVER EXPRESSED FOR SUN 1. SOME OVER COMPENSATION FOR OVER EXPRESSION OF SUN 1. AND THE ONES THAT OVER OVER COMPENSATE, RESORT IN THESE DISTORTED NUCLEI AND END UP BEING THE PATHOLOGICAL CELLS. AND SO, IF ONE IS ABLE TO USE siRNA TO KNOCK THEM DOWN, THEN WHAT YOU FIND IS THAT THE CELLS ARE NOT KNOCKED DOWN AND REMAIN CREAMIATED BUT ALL THE CELLS IN HTPS ARE KNOCKED DOWN FOR SUN 1 AND NOW RECOVER THE NORMAL NUCLEAR MORPHOLOGY AND WE PREDICT THAT THOSE CELLS WOULD BE PHYSIOLOGICALLY NORMAL AND IT’S THE OVEREXPRESSION THAT ENDS UP IN THE BAD ACTOR. SO, WHAT WE THINK IS HAPPENING IN DISTROPEIC DISEASES, AT LEAST FROM THE EXAMPLE THAT WE HAVE WORKED ON, IS THAT WHEN YOU HAVE A MUTATION IN ONE ENTITY, THERE IS COMPENSATORY OVERSYNTHESIS OF PROTEINS OF OTHER ENTITIES. AND IN THIS CASE, WHEN YOU HAVE A MUTATION IN LAMIN A, YOU HAVE OVERSYNTHESIS WITH SUN 1 PROTEIN AND IT’S THE OVER SYNTHESIS, PER SE WITHOUT BEING GETTING RID OF THEM, THAT ENDS UP DISTORTING THE NUCLEUS AND CREATING THE PATHOLOGY. SO THEREFORE, IT’S FULLY IN KEEPING WITH THE PAPER THAT WAS RECENTLY PUBLISHED IN SCIENCE TRANSLATIONAL MEDICINE FROM DR. COLLINS LAB, WHICH SAYS THAT IF YOU NOW INCREASE THE PROTEIN DEGRADATION PATHWAY, WHICH RAP MICE IN IS AN INDUCER OF THE PATHWAY, WHICH GETS RID OF THESE OVER ACCUMULATED PROTEIN AGGREGATES, THEN TREATMENT WITH THAT DRUG IS CONSISTENT AND RETURNING BACK TO NORMALCY. AND WE THINK THAT ABNORMAL AGING AND NORMAL AGING, MAY BE MECHANISTICALLY THE SAME. BECAUSE AS YOU KNOW, THERE IS ALSO A BODY OF LITERATURE THAT SAYS, IF YOU TREAT MICE, NORMAL MICE, WITH RAPAMYCIN, WHICH OF COURSE ALSO IMPROVES THEIR JUNK PROTEIN DEGRADATION PATHWAY, THOSE MICE ACQUIRE HIGHER INCREASED LONGEVITY. ANOTHER WAY OF LOOKING AT THE HYPOTHESES IS THAT HERE IS THE NORMAL CELL. THIS IS WHAT HAPPENS TO THE CELL THAT IN FACT IS KNOCKED OUT FOR LAMIN. SO SOMEONE OVERACCUMULATES AND THEN THIS IS WHAT HAPPENS TO THE CELL WHEN YOU HAVE KNOCKOUT OF LAMIN AND KNOCKOUT OF SUN 1. THESE OVERACCUMULATION GOES AWAY AND SO DO ALL THE BAD THINGS THAT HAPPEN WHEN THE NUCLEUS GETS DISTORTED BY HYPERACTIVE DNA DAMAGE AND ABNORMAL TRANSCRIPTION AND CYTOTOXICITY FROM OVER ACCUMULATION. THEY ALL BECOME AB SOLVED AND THEREFORE YOU TURN BACK TO NORMAL LONGEVITY IN MOST OF THESE MICE. I THINK IN MANY WAYS, WHEN YOU DO GOOD SCIENTIFIC FINDINGS, THEY SORT OF END UP CONVERGING AND DOVETAILING TOGETHER, WHICH IS THAT WE ALSO KNOW THAT AGING ALSO RESULTS IN WEAKENING OF THE CHECKPOINT AND AGING ALSO RESULTS IN INCREASING ANEUPLOIDY. AND SO MY GUESS IS THAT SOMEWHERE DOWNSTREAM, WHEN YOU LOOK AT SOME OF THESE PREMATURE AGING MICE, WILL YOU FIND THAT THEY HAVE INCREASED PROPENSITY FOR ANEUPLOIDY WITH THOSE NUCLEAR DISTORTIONS AND WITH ALSO DNA DAMAGE AND MY GUESS IS THAT SOMEWHERE, IF THEY LIVED LONG ENOUGH, WE WOULD HAVE FOUND THEY HAVE WOULD INCREASED PREVALENCE OF TUMORIGENESIS. SO, LET ME WRAP IT UP BY SUMMING FOR YOU THAT IN THE FIRST HALF OF MY TALK, I TALKED TO YOU BASICALLY ABOUT HTLV1 TRANSFORMATION AND OLD YOU ONLY A LITTLE SNIPPET OF WHAT OUR REAL PROGRAM HAS BEEN. SO I TOLD YOU ONLY ABOUT THIS PARTICULAR ASPECT OF OUR RESEARCH. IN FACT, THERE ARE ALL THESE DIFFERENT OTHER AREAS WHICH ALL OF THESE DIFFERENT POSTDOCS OVER THE YEARS HAVE PARTICIPATED IN. ALL OF THESE OTHER STEPS ARE ALSO IMPORTANT LIKE GENERATION OF REACTIVE OXYGEN SPECIES, OF ALSO ACTIVATION OF MICRORNAs AND SO ON, THAT I HAVEN’T HAD TIME TO TOUCH UPON. AND SO I THINK THAT THIS IS VERY EXCITING FOR US. THIS IS I’M VERY GRATEFUL TO GEORGE FOR SORT OF STARTING ME ON THIS PARTICULAR SYSTEM, AND WE THINK WE HAVE MADE GOOD MILEAGE ON THE ROAD TRAVELING TO TRY TO UNDERSTAND HOW THE VIRUS CAUSES LEUKEMIA. WELL AND BY-AND-BY, I THINK THAT THE WONDERFUL COMMUNITY OF DOING SCIENCE AT THE NIH IS THAT WE ARE ABLE TO PURSUE INTERESTING LEADS AND I SIMPLY WANT TO SUM UP THE SECOND HALF OF MY TALK BY SORT OF HIGHLIGHTING THIS PARTICULAR NEWS AND VIEWS ARTICLE THAT WAS WRITTEN IN BIOLOGY WHEN OUR PAPER WAS PUBLISHED. YOU CAN SUM UP THE ISSUE HERE, AS MOTHER WOULD ALWAYS TELL YOU, TOO MUCH SUN IS REALLY BAD FOR YOU. AND SO, IN OUR CASE, TOO MUCH SUN WILL REALLY MAKE YOU AGE FASTER AND CERTAINLY DO BAD THINGS TO YOUR PHYSIOLOGY. NOW, LET ME CLOSE WITH THIS SLIDE TO ACKNOWLEDGE CURRENT POSTDOCTORAL FELLOWS IN LAB. PREVIOUS ONES WHO WORKED IN THE LAB WHO CONTRIBUTED TO SOME OF THE STORIES THAT I TOLD YOU TODAY. SOME OF OUR COLLABORATORS ARE THE LABORATORIES AND NIAID INTRAMURAL FUNDING AND OVER THE LAST 18 YEARS OR SO, I RECEIVED FUNDING AND WE HAVE ALSO HAD A CHALLENGE GRANT THAT WAS AWARDED TO US FROM BILL AND MELINDA GATES FOUNDATION. I WANT TO CLOSE BY MAKING A REMARK ABOUT THREE THINGS THAT GEORGE TAUGHT ME SEPARATE FROM SCIENCE, WHICH I WOULD REALLY LIKE TO ACKNOWLEDGE. GEORGE TAUGHT ME THREE THINGS VERY IMPORTANT. HE SAYS ALWAYS REMEMBER YOUR FAMILY, FRIENDS AND YOUR COMMUNITY. AND SO, I WANT TO CLOSE BY ACKNOWLEDGING OF COURSE, THIS IS FAMILY PICNIC THAT THE SOCIETY THAT I WAS PRIVILEGED TO LEAD WITH THE CHAPTER AT THE NIH, SOCIETY OF CHINESE BIOSCIENTISTS IN AMERICA. THIS A FAMILY PICNIC WE ORGANIZE EVERY YEAR, EVERY SUMMER. THIS WAS AT THE CABIN JOHN. AND IT’S A REMARKABLY WONDERFUL OCCASION FOR US TO SORT OF RECOGNIZE THE IMPORTANCE OF THE FAMILY IN TERMS OF ALLOWING US TO DO THE LONG HOURS AND SORT OF THE COMMITTED WORK WE DO AT THE NIH. AND FINALLY I WANT TO GIVE A FEW WORDS OF THANKS TO OF COURSE MANY OF YOU. MOST OF YOU ARE MY FRIENDS SITTING IN THE AUDIENCE AND I REALLY APPRECIATE THAT YOU HAVE COME. BUT WITHIN OUR LARGER NIH COMMUNITY AS I MENTIONED, THERE IS A SMALLER COMMUNITY OF CHINESE BIOSCIENTISTS WHO I MEET WITH EVERY MONTH AND IN MANY WAYS, WE COLLABORATE AND WE OFFER EACH OTHER ADVICE AND SUPPORT. AND SO, SOME INDIVIDUALS THAT I WOULD PARTICULARLY LIKE TO RECOGNIZE, YOU ALL KNOW PAUL, YOUNG, KJ, JAY, SHING AND A NUMBER OF OTHER INDIVIDUALS HERE. AND THIS WAS LAST YEAR WHEN OUR SOCIETY AND OUR CHAPTER ORGANIZED TOGETHER WITH NIAAA SYMPOSIUM IN BUILDING 31. MANY OF THESE INDIVIDUALS OVER THE LAST 27 YEARS HAVE PROVIDED REALLY IMPORTANT GUIDANCE AND SUPPORT TO ME FOR MY CAREER AT THE NIH AND FOR ALL OF YOU, FOR MY FAMILY AND FOR ALL THE FRIENDS, MANY OF YOU IN THE AUDIENCE AND CERTAINLY FOR THE GREATER NIH COMMUNITY, THOSE ARE THE THINGS THAT GEORGE HAS TAUGHT ME THAT ARE IMPORTANT AND I THANK YOU. [ APPLAUSE ]>>ALTHOUGH THE HOUR IS A BIT LATE, I THINK GEORGE WOULD NOT REST EASY IF WE DIDN’T HAVE SOME QUESTIONS. TOM, YOU HAVE A QUESTION?>>– THE SECOND HALF OF YOUR LECTURE IS AGING AS EARLY SUNSET, I WOULD LIKE TO TURN TO TAX AND ANEUPLOIDY. THROUGH THE COURSE OF HTLV1 INFECTION ARE WHICH FOR MANY PEOPLE IS LIFELONG, THAT IS IT STARTS AT BIRTH, ONE HAS A CARRIER STATE WHERE TAX IS EXPRESSED. DURING THAT PERIOD, ARE THE — IS THERE INCREASED ANEUPLOIDY? AND AT THE OTHER END OF THE DISEASE, MOST — OVER HALF OF THE PATIENTS WITH ACUTE ADULT T-CELL LEUKEMIA, NO LONGER EXPRESS TAX. THEY EXPRESS HTZ BUT DOES THIS ANEUPLOIDY CONTINUE IN THAT PHASE IN THE ABSENCE OF TAX?>>SO, LET ME ANSWER THE SECOND PART FIRST. SO, WE THINK THAT IN THE CASE OF ATL, IT IS A LITTLE BIT DIFFERENT FROM OTHER VIRAL CAUSE TUMORS. SO FOR EXAMPLE, I OCCASIONALLY SERVE ON STUDY SECTIONS ON VIROLOGY STUDY SECTIONS AND I WOULD HAVE THIS RUNNING ARGUMENT WITH HPV VIRALLOLOGISTS AND THEY ARGUE IN THEIR CASE, IF YOU LOSE E6 AND E7, YOU CAN NEVER MAINTAIN THE CERVICAL CANCER. IN OUR CASE, AS YOU KNOW, WE LOSE TAX, AND THE CELLS ARE HAPPY TO BE ATL CELLS. SO I THINK WHAT HAPPENS IS THAT YOU KNOW, CAN, CHEMICAL INDUCED CARCINOGENESIS HAS NO ONCOGENE. I MEAN NO VIRAL ONCOGENE. IT IS SIMPLY CHANGING GENETIC COMPLIMENT OF THAT PARTICULAR GENOME. SO I THINK YOU REACH A CERTAIN STAGE IF HAVE YOU ANEUPLOIDY, YOU HAVE ACQUIRED THAT ALL OF THE INDEPENDENT PROLIFERATION AND ANTIAPOPTOSIS AND ANTISI NECESSARIENCE SIGNALS. SO YOU NO LONGER NEED TAX. YOUR FIRST QUESTION IS, IN EARLY INFECTION, DOES ANEUPLOIDY OCCUR? AND HOW DO WE MEASURE THAT? I HAVE TO PROFESS, WE DON’T HAVE EVIDENCE, SO WE CAN’T TELL YOU. BUT I CAN SAY THAT THERE ARE OR THERE IS A STRUGGLE WHEN THE VIRUS INFECTS A CELL BECAUSE AS YOU KNOW, THE FIRST CELL OF DEFENSE AND CELLS ARE INCREDIBLY MORE ALTRUISTIC THAN YOU AND I ARE. CELLS ARE WILLING TO COMMIT SUICIDE BEFORE ALLOWING ANOTHER CELL TO BECOME INFECTED. SO I THINK THE FIRST SIGNAL AND THE FIRST INTENSE OF THE CELL WHEN IT IS INFECTED BOY A VIRUS, WOULD BE TO COMMIT APOPTOSIS OR IN SOME OF JOE’S STUDIES, THE CELL WOULD COME IN SI NECESSARIENCE AND GO TO SLEEP TO PROTECT AGAINST THE SPREADING OF INFECTION. SO THERE IS A SELECTION OF ONLY CELLS THAT EXPRESS A SMALL AMOUNT OF THE TAX PROTEIN. THOSE CELLS, I THINK WILL NOT GIVE YOU WHOLESALE ACQUISITION OF ANEUPLOIDY BUT SLOWLY YOU WILL GAIN TRY 70 AND 21 AND THAT KIND AND THEN THE THINGS START ROLLING OVER TIME. AND SO I THINK THE REALLY TRAUMATIC CHANGES, THOUGHT APOPTOSIS, AND SO YOU GET THE MILD GUYS AND THEY BECOME SORT OF UNITED FROM WHICH ACCUMULATIONS OCCUR AND THEN FINALLY WHETHER YOU HAVE SUFFICIENT ACCUMULATION, YOU BECOME NOW TAX INDEPENDENT. 90% OF THAT IS CONFABALATION. I THINK 10% IS PROBABLY ROUTED IN OBSERVATIONS. DAVID?>>HI, THAT WAS A TERRIFIC TALK. SO, FROM THE POINT OF VIEW OF THE VIRUS, BECAUSE PRESUMABLY TAX EVOLVED THIS FOR THE VIRAL LIFE CYCLE, NOT FOR THE SAKE OF TUMORS. WHAT THE IS ADVANTAGE TO THE VIRUS FOR THE INTERACTION WITH MAD 1? I THINK THE VALUE FOR THE VIRUS IS THAT THE VIRUS — SO I THINK THE VIRUS IS BASICALLY PRO PROLIFERATIVE AND TRANSFORMATION IS A PERHAPS UNWELCOME BUT UNAVOIDABLE SIDE EFFECT OF WHENEVER YOU INCREASE PRO PROLIFERATION. BECAUSE THIS IS A VIRUS THAT DOES NOT PRODUCE A BIRTH SIZE OF VIRAL PARTICLES. SO THE ONLY WAY TO AMPLIFY THE VIRAL GENETIC MATERIAL IS THAT YOU MAKE THE INTEGRATEED PROVIRUS IN A CELL THAT CARRIES INTEGRATED PROVIRUS DIVIDE MORE AND MORE AND MORE. SO IT’S VERY DIFFERENT FROM HIV. IT DOESN’T RELEASE THE BURST OF VIRAL PARTICLES. IT SIMPLY MAKES OR DRIB ELSE OUT A VIRUS. SO IT BEHOOVES THE VIRUS TO REALLY THE SMARTEST HTLV WOULD BE A HTLV THAT IMMORTALIZES T-CELLS WITHOUT GOING OVER THE TIPPING POINT OF MAKING INTO CANCERS. SO HISTORICALLY, AS YOU KNOW, MANY CANCER VIROLOGY STUDY TRANSFORMATION IN-VITRO AND WE ALWAYS THINK ABOUT THE FIRST STEP IS IMMOBILIZATION AND THEN THAT IS BY TRANSFORMATION, IMMORTALIZED CELLS, PROLIFERATE IN FINITE IMBUT CANNOT CAUSE TUMORS IN VIVO. BUT WHEN YOU TIP TO THE TRANSFORMATION STAGE, NOT ONLY CAN YOU REPLICATE FOREVER, YOU CAN ALSO CAUSE TUMORS WHEN YOU INJECT INTO ANIMALS. SO I THINK HTLV, UNFORTUNATELY HAS EVOLVED TO BE OVERLY ROBUST. THE BEST HTV WOULD BE IMMORTALIZING VIRUS BUT NOT A TRANSFORMING VIRUS. BUT I THINK IN THE QUEST FOR EFFICIENT IMMORTALIZATION, THE VIRUS HAS ALWAYS ACQUIRED THE ABILITY TO TRANSFORM.>>I’M STILL TRYING TO SQUARE YOUR MORPHOLOGICAL OBSERVATION OF BINDING OF LABEL TAX TO SPINDLE PROTEINS. MECHANISTIC STUDIES. YOU SAID ANEUPLOIDY SPINDLE PROTEINS ARE FRAGMENTED ADIPOSE BUT THE TAX IS NONETHELESS ABLE TO BIND TO THIS POLINGS WHICH HAVE RISEN OUT OF FRAGMENTED STARTING POINT OF PRECURSORS. DOES THAT MEAN THAT THE TAX CAN BIND NONSPECIFICALLY TO THE CONFIDENCE OF THE POLE? OR THAT THE POLES WHICH ARE ASSEMBLED REGARDLESS, THEY ARE COMING FROM FRAGMENTEDDED PARTS AND THEY ARE ALL COMPETENCE TO BIND THE TAX?>>SO THE SIMPLE EXPLANATION WOULD BE TAX IS DIRECTLY ABLE TO BIND PARACENTRUM PROTEIN. AND THEREFORE, EVERY TIME WHERE YOU SEE PARASEN TRIN, YOU WILL FIND TAX. AND WHICH IS WHAT THE STAINING TELLS: WHEN TAX BINDS ON THE PAIR CENTRUM, IT TENDS NOT TO BE ABLE TO FORM THE BIPOLAR PROGRAM. INSTEAD TENDS TO FORM MULTI-POLAR PROGRAM. IT’S DIFFERENT FROM PARACENTRIM. A DIFFERENT TARGET.>>OKAY. I THINK WHEN GEORGE STARTED WORKING ON SB40 HIS THOUGHT WAS THAT A SIMPLE VIRUS WHICH HAD SUCH A PROFOUND EFFECT ON CELL BIOLOGY WOULD ALLOW US TO LEARN A LOT ABOUT HOW THE CELL WORKS AND I THINK GEORGE WOULD BE EXTREMELY PLEASED TO KNOW HOW FAR THIS WORK HAS COME.

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