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0:07
>> OKAY, THANKS FOR COMING. LET'S GET STARTED. I AM MIA SUN. IT'S MY PLEASURE TO INTRODUCED TO'S SPEAKER. BUT BEFORE I FORGET, TODAY'S CME CODE IS 5016. -- 50116. AND THE PEOPLE WATCHING THE TALK ON VIDEOCAST CAN SUBMIT YOUR QUESTIONS ON THE BUTTON SEND LIVE FEEDBACK. YOU CAN ASK THE SPEAKER QUESTIONS THAT WAY. OKAY, SO LET ME TRY TO GIVE A GOOD INTRODUCTION. BECAUSE IT'S REALLY NOT AN EASY JOB TO CONDENSE WENDELL LIM'S ACCOMPLISHMENT INTO A SHORT INTRODUCTION BUT I WILL DO MY
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BEST. SO WENDELL LIM, DR. WENDELL LIM IS BUYER'S DISTINGUISHED PROFESSOR AT U.C. SAN FRANCISCO, SCIENTIST OF MOLECULAR, HE GOT BACHELOR'S HARVARD IN CHEMISTRY. POSTDOC AT YALE. ALSO IN BIO PHYSICS AND BIOCHEMISTRY. HE JOINED U.C. SAN FRANCISCO IN 1996. SINCE THEN HE HAS TAKEN NUMEROUS LEADERSHIP POSITIONS INCLUDING THE DIRECTOR OF U.C. SAN FRANCISCO CENTER FOR SYSTEMS AND SYNTHETIC BIOLOGY, ONE OF THE NIH CENTERS IN SYSTEMS BIOLOGY UNTIL 2019. SINCE 2020 HE HAS BEEN THE
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DIRECTOR OF CELL DESIGN INSTITUTE, AND ALSO DIRECTOR OF CENTER FOR SYNTHETIC IMMUNOLOGY, BOTH AT U.C. SAN FRANCISCO. AND BEYOND U.C. SAN FRANCISCO HE HAS BEEN REALLY SERVING THE ENTIRE COMMUNITY OF PEOPLE WHO ARE DOING QUANTITATIVE SYSTEMS BIOLOGY AND SYNTHETIC BIOLOGY IN THIS ANNUAL CONCERT SERIES WE CALL WINTER Q BIO. A LONG TIME CO-ORGANIZER IN THAT CONFERENCE SERIES. WENDELL IS REALLY A PIONEER IN SYNTHETIC BIOLOGY APPROACHES AND HE HAS OPENED MANY MULTIPLE
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DIFFERENT RESEARCH AVENUES. SO I WILL JUST TRY TO COVER A LITTLE BIT HERE. SO HIS APPROACHES TO ENGINEERING CELLS IS REALLY COOL IN THE SENSE YOU BASICALLY TELL LYMPHOCYTES TO TELL INSTRUCTIONS LIKE IF YOU SEE A AND B, DO, C AND D. SUCH AS KILLING THE TARGET CELLS OR ACTIVATE SOME GENES. SO THESE ARE REALLY HIGH PRECISION THERAPEUTIC APPROACHES HE HAS PIONEERED. AND IN 2016 HE PUBLISHED NOT ONE, BUT THREE CELL PAPERS THAT REALLY DEMONSTRATE THAT THIS WORKS WITH THE SYSTEM THAT HE
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TERMS SYNTHETIC NOTCH RECEPTORS, ALSO CALLED SYN NOTCH SYSTEMS. AND IN 2022 HE PUBLISHED TWO PAPERS IN SCIENCE. ABOUT HOW CAR T-CELL PHENOTYPES CAN BE MANIPULATED BY MIXING AND MATCHING OF DIFFERENT MOTIFS OR DOMAINS. REALLY BEAUTIFUL WORK. HE SO FAR PUBLISHED MORE THAN 150 PEER-REVIEWED PAPERS IN HIS OVER 60 PATENTS. AND I WAS CURIOUS TO SEE WHAT IS THE TOP-SIDED PAPER OUT OF ALL OF THIS. SO I FOUND IT'S ACTUALLY THIS CELL PAPER PUBLISHED IN 2013. TOGETHER WITH ACTUALLY ANOTHER
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WALS SPEAKER NEXT MONTH, JENNIFER DOUDNA, SHOWING CRISPR CAN BE MADE TO BE REVERSIBLE. HE DEVELOPED THIS CRISPR EYE METHOD IN THE PAPER. FOR ALL HIS ACCOMPLISHMENTS HE HAS RECEIVED NUMEROUS HONORS AND AWARDS, MANY DISTINGUISHED LECTURES. HE HAS BEEN EDITORIAL BOARD OF CELL AND SCIENCE. HOWARD HUGHES MEDICAL INSTITUTE INVESTIGATOR. AND EVEN BACK IN 2012, HE WAS NAMED ONE OF THE 50 PEOPLE WHO WILL CHANGE THE WORLD BY U.K. MAGAZINE WIRED. AND TODAY'S LECTURE, WE WILL
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SEE HOW HE CONTINUES TO REALLY PUSH THE BOUNDARIES, BOTH TECHNOLOGICALLY AND CONCEPTUALLY AT THE FOREFRONT OF SYNTHETIC BIOLOGY. THANK YOU SO MUCH FOR VISITING NIH. WENDELL, I WOULD LIKE TO WELCOME YOU. [ APPLAUSE ] >> WENDELL LIM: THANK YOU. CAN EVERYONE HEAR ME OKAY? SO THANK YOU VERY MUCH, MIA, FOR THAT KIND INTRODUCTION AND FOR INVITING ME. AND HELLO TO EVERYONE BOTH HERE IN PERSON AND ONLINE. SO WHAT I WOULD LIKE TO TALK ABOUT IS OUR EFFORTS OVER THE
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LAST SEVERAL YEARS TO TRY TO ASK WHETHER WE CAN ACTUALLY CREATE NEW BIOLOGY. WHAT DOES IT TAKE TO ACTUALLY LEARN HOW TO PROGRAM CELLS AND CELLS IN THEIR FUNCTION AND HOW CAN WE APPLY THIS TO VARIOUS IMPORTANT THERAPEUTIC PROBLEMS. HERE ARE MY DISCLOSURES. AND THEN WHAT I WOULD LIKE TO DO IS START BY TALKING ABOUT THIS GENERAL FIELD OF SYNTHETIC BIOLOGY AND HOW I THINK ABOUT IT. AND IT'S REALLY THIS IDEA OF LEARNING BY BUILDING. I THINK THAT OVER THE LAST MANY
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DECADES WITH THE ADVENT OF MOLECULAR BIOLOGY AND UNDERSTANDING THE GENE, WE HAVE COME TO REALIZE THAT BIOLOGY IN THE HIGH-LEVEL FUNCTIONS WE SEE IN VARIOUS ORGANISMS ARE THE RESULT OF A HIERARCHY OF LEVELS WHERE GENETIC PROGRAMS LEAD TO ENCODE MOLECULAR NETWORKS LEAD TO CELLULAR NETWORKS AND IT'S REALLY THE ENSEMBLE OF THESE THAT LEAD TO THE FUNCTIONS WE ARE SOEN NAM ORED OF THAT WE SEE IN LIVING SYSTEMS. RIGHT NOW IN GENERAL, THE WAY WE LOOK AT THESE SORTS OF
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SYSTEMS WE ARE INTERESTED IN PARTICULAR FUNCTION AT A HIGH LEVEL AND WE TEND TO TRY TO DECONSTRUCT AND MAP OUT THE GENES AND HOW DO THEY INTERACT WITH ONE ANOTHER. THIS IS FOCUSED ON TAKING THOSE BIOLOGICAL BEHAVIORS THAT EXIST AND TRYING TO UNDERSTAND THEM. IN A WAY, WHAT WE ARE INTERESTED IN SYNTHETIC BIOLOGY, WE ARE INTERESTED IN THAT PROBLEM BUT REALLY IN A BIGGER CONTEXT. MAYBE ANOTHER WAY OF PHRASING WHAT ARE POSSIBLE. WHAT ARE THINGS YOU COULD BUILD WITH BIOLOGICAL SYSTEMS?
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AND THAT IS IMPORTANT FROM SEVERAL DIFFERENT PERSPECTIVES. ONE IS THAT REALLY, YOU KNOW, IF YOU LOOK AT A BROADER SET OF SYSTEMS AND UNDERSTAND WHAT CAN ACHIEVE A CERTAIN FUNCTION, WHAT ARE THE GENERACIES, PAERNTS, YOU CAN GET A BETTER UNDERSTANDING OF THE RULES THAT GOVERN THE BEHAVIOR. IF YOU EXPLORE THIS SPACE OF USING BIOLOGIC COMPONENTS TO CREATE NEW SYSTEMS PROBABLY THERE ARE INTERESTING NEW BEHAVIOR S OUT THERE SOME WHICH COULD BE THERAPEUTICALLY USEFUL.
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WE ARE INTERESTED IN BOTH QUESTIONS AND EXPLORING THEM REALLY IN CONJUNCTION. ONE WAY OF THINKING ABOUT THAT, THE TRADITIONAL WAY, PERHAPS THINKING ABOUT PROBLEMS IN IMMUNOLOGY OR DEVELOPMENT, THESE ARE BOTH CASES WHERE SYSTEMS OF CELLS DO AMAZINGLY COMPLEX AND PRECISE ACTIONS. PERHAPS TRYING TO REPHRASE THIS IN A MORE GENERATIVE WAY IN TERMS OF HOW YOU CREATE FUNCTION FROM THE BOTTOM UP. WE COULD ASK IF WE UNDERSTAND THE DESIGN LOGIC OF THESE SYSTEMS HOW MIGHT WE BE ABLE TO
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FOR INSTANCE IN IMMUNOLOGY CREATE IMMUNE CELLS THAT KILL FOREIGN CELLS THAT NATURALLY INVADE. OR SYNTHETIC DEVELOPMENT HOW CAN WE PROGRAM CELLS SO THEY SELF ORGANIZE IN CUSTOM TISSUES FOR REGENERATION OR REPAIR. BY TAKING ON THESE SORTS OF PROBLEMS WE ARE TRYING TO DO THING THAT'S ARE USEFUL AND ONLY CELLS COULD DO BUT ALSO COULD HELP US GIVE US BIGGER CONTEXT FOR UNDERSTANDING THE PRINCIPLES AND RULES AND LOGIC OF BIOLOGICAL REGULATION. THE OTHER THING IS RIGHT NOW WE
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ARE VERY FOCUSED ON THE NOTION OF TRYING TO CHANGE CELLULAR CONVERSATIONS. HOW CELLS INTERACT WITH ONE ANOTHER AS NETWORKS. OF COURSE BIOLOGY OCCURS AT MANY DIFFERENT LEVELS. ONE OF THE POINTS I WANT TO MAKE, WE HAVE MADE AMAZING ADVANCES AT THE ATOMIC AND MOLECULAR SCALE UNDERSTANDING THE INTERACTIONS BETWEEN DIFFERENT MODULES. FOR EXAMPLE, THE FIELD OF CHEMISTRY USED TO BE ABOUT STUDYING INDIVIDUAL MOLECULES. BUT REALLY NOW WE HAVE THIS GREAT FUNDAMENTAL UNDERSTANDING
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HOW ORBITALS AND BONDS WORK. AND REALLY CHEMISTRY IS MUCH MORE BASED ON PRINCIPLES AND NOT ABOUT WHAT MOLECULES DON'T AND DO EXIST BUT WHAT CAN EXIST. LIKE THAT WE UNDERSTAND MOLECULES AT A BETTER LEVEL. BUT I THINK A LOT OF BIOLOGY WE LOOK AT THE HIGHER LEVEL BOILS DOWN TO MOLECULES ENGAGING AT THE CELLULAR LEVEL. WE ARE INTERESTED IN TRYING TO FOCUS ON THAT. WHAT WE WOULD LIKE TO BE ABLE TO DO IS TO DEVELOP THE FIELD OF CELL ENGINEERING AND CELL DESIGN TO BE AS FACILE WHAT WE
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CAN DO WITH TODAY'S SYNTHETIC CHEMISTRY OR PROTEIN ENGINEERING. THEY HAVE AMAZING CAPABILITIES I THINK MOLECULES BY THEMSELVES CAN'T HAVE, WHETHER SMALL MOLECULES OR BIOLOGICS. MOLECULES COULD HAVE PRECISE SPECIFICITY. BECAUSE THESE ARE MOLECULES THEY WILL GO EVERYWHERE, AND THE SAME GENES ARE EXPRESSED IN DIFFERENT CELLS YOU WILL HAVE SYSTEMIC CROSS REACTIONS AND TOXICITIES. CELLS, HOWEVER, THIS HAPPENS OF COURSE NATURALLY IN THE BODY, CELLS HAVE THE ABILITY TO SENSE
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AND INTEGRATE ENVIRONMENTAL INFORMATION TO CONTROL WHERE THEY GO AND WHEN THEY EXECUTE CERTAIN FUNCTIONS. CELLS CAN MIGRATE. THEY CAN ADHERE OR BE RETAINED IN CERTAIN PLACES. INTERACT SPECIFICALLY AND LOCALLY INDUCE ACTIVITIES, PROLIFERATION, PAYLOADS OR COMMUNICATION WITH OTHER CELLS. I WOULD ARGUE MANIPULATING CELLULAR AGENTS MAY BE A FAR MORE POWERFUL WAY TO INTERACT WITH DISEASES SPECIFICALLY COMPLEX DISEASES THAT INVOLVE CELLULAR ECOSYSTEMS LIKE CANCER, AUTOIMMUNE ORFEI BREAUX
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SIS. WHAT I WOULD LIKE TO TALK ABOUT TODAY QUICKLY IS THREE THINGS. ONE I WANT TO GIVE YOU AN INTRODUCTION TO THE TYPE OF SYNTHETIC BIOLOGY TOOLKIT TO REWIRE CELLS. WHAT ARE THE FUNDAMENTAL UNITS IF YOU WERE GOING TO ENGINEER CELLS WHAT ARE THE MODULES YOU NEED THAT WOULD GIVE YOU THE MOST CAPABILITIES. I WILL TELL YOU ABOUT SOME THINGS WE HAVE BEEN WORKING ON AND REWIRE HOW CELLS INTERACT WITH THEMSELVES AND THEIR ENVIRONMENT. AND THEN I WANT TO TALK ABOUT
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HOW WE CAN THEN APPLY THIS TOOLKIT TO TWO DIFFERENT PROBLEMS. MOSTLY I WILL TALK ABOUT DEVELOPING NEXT GENERATION IMMUNE CELL THERAPIES FOR SOLID TUMORS BUT BEYOND CANCER, INCREASED RECOGNITION, THIS DEVELOPMENT OF A TISSUE GPS SYSTEM THAT ALLOWS US TO TELL CELLS TO GO TO AND ONLY ACT IN A CERTAIN ANATOMIC ISSUE AND PROGRAMMING CELLS TO TREAT DISEASES LIKE CANCER LIKE AUTOIMMUNITY OR NEUROINFORMATION. AND A SHORTER PART TO TALK ABOUT EFFORT TO APPLY SAME TOOL
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WITH DIFFERENT PROBLEM THAT OF DEVELOPMENT TRYING TO UNDERSTAND HOW WE CAN WIRE CELLS TO SELF-ORGANIZE AND SELF PATTERN INTO VARIOUS ARCHITECTURES AND HOW THIS MIGHT BE USED TO HELP GUIDE STEM CELLS TO DEVELOP INTO SPECIFIC TISSUES OR SHAPES. SO LET ME FIRST TALK ABOUT THIS TOOLKIT FOR REWIRING CELLULAR NETWORKS. LET'S SAY WE HAVE A PROBLEM OF A CAR-T CELL AND WE WANT TO ENGINEER TO TREAT A SOLID TUMOR. WE NEED IT TO BE ABLE TO TALK TO THE TUMOR, RECOGNIZE IT,
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HAVE A CONVERSATION, TALK TO THE STROMA, AS WELL AS OTHER IMMUNE CELLS IN THE ENDOGENOUS IMMUNE SYSTEM BUT ALSO AMONGST THEMSELVES IN AUTOCRATIC SORT OF WAYS. IF WE WANT TO DO THAT WHAT ARE THE WORDS WE NEED TO WRITE NEW CELLULAR PROGRAMS TO CHANGE THE RELATIONSHIPS AND CONVERSATIONS THESE CELLS HAVE, IDEALLY WE WOULD LIKE TO, LIKE WITH THIS DRY ERASE MARKER PENCIL IN NEW THINGS AND ASK HOW DOES THAT CHANGE THE BEHAVIOR OF THE NEW SYSTEM. WE WANT TO BE ABLE TO DO THAT
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AND WE WANT TO HAVE GENETICALLY ENCODED COMMANDS THAT COULD PROGRAM NEW CELLULAR INPUT/OUTPUT RELATIONSHIPS. THIS IS STILL DIFFICULT BUT FINITE QUESTION, WE WOULD LIKE A CELLULAR NODE TO ENGINEER TO DETECT A NEW INPUT LIKE XYZ, SOMETHING OUTSIDE ITS ENVIRONMENT, HAVE THAT TURN ON A NUMBER OF CANONICAL RESPONSES, PRODUCING SECRETED SIGNAL OR RECEPTOR TO SEND SOMETHING OR CHANGE SHAPE OR DIVIDE AND GROW. CLASSICAL RESPONSES WE WOULD LIKE TO BE ABLE TO HAVE AS OUTPUTS.
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SO WHAT ARE SOME OF THESE GENETIC COMMANDS? MANY OF YOU ARE PROBABLY FAMILIAR WITH CHIMERIC -- CAR. BASICALLY IT TAKES THE INTRACELLULAR PARTS OF THE TCR COMBINED WITH SIGNATORY RECEPTORS BUT PUTS ON THE OUTSIDE NEW ANTIBODY FOR A PARTICULAR ANTIGEN. THAT ALLOWS US TO REDIRECT TO A CELL THAT HAS THIS NEW ANTIGEN. SO IT SHOWS HOW YOU CAN REPROGRAM THAT OUTPUT TO LINK IT TO ENDOGENOUS OUTPUT. HERE ARE THINGS WE HAVE DEVELOPED AND ARE WORKING ON, ONE I WILL GO INTO MORE DETAIL,
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SYNTHETIC NOTCH SYSTEM. THAT'S THE WAY WE COULD CHANGE THE OUTSIDE WITH SINGLE CHAIN ANTIBODY, WHAT IT DOES, IT RELEASES TRANSCRIPTION ALDO MAIN THAT GOES INTO THE NUCLEAL DOMAIN. SO IT'S A NEW WAY TO PUT A NEW INPUT BUT HAVE ALMOST ANYTHING AS AN OUTPUT THAT WE CAN ENCODE GENETICALLY. IT'S VERY, VERY FLEXIBLE AS AN INFORMATION TRANSMISSION MODALITY. AND THEN SOME OTHER THINGS, WHAT I WILL CALL SYNTHETIC CELL ADHESION MOLECULES, SYNCAMS. WHAT KIND OF INTERFACE BY
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CHANGING THE INTRACELLULAR DOMAINS AND CHANGE THE CYTOSKELETON. ENGINEERED CHEMOKINE SYSTEM. GIVE YOU LONG-TERM MEMORY AND FATE BIFURCATION. I WON'T TALK ABOUT TODAY. LET'S GO OVER THE SYN NOTCH SYSTEM. THE NATURAL NOTCH RECEPTOR DETECTS SIGNAL DELTA ON A NEIGHBORING CELL, THIS MECHANICAL FORCE LEADS TO CLEAVAGE AND INTRACELLULAR KETS CUT OFF AND REGULATES TRANSCRIPTION OF VARIOUS GENES. WE COULD RECOGNIZE ARBITRARY MOLECULE X ON THE OUTSIDE WORLD. PUT TRANSCRIPTION FACTOR ON THE
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INSIDE. THIS COULD TURN ON WHATEVER GENES, WHY WE PUT BEYOND THAT RESPONSIVE PROMOTER. SO THIS ALLOWS US TO CHANGE WHAT WE DETECT IN THE ENVIRONMENT AND CONTROL ALSO WHAT THE CELLULAR RESPONSE IS. MOREOVER, WE HAVE SHOWN THAT WE CAN CHANGE THE OUTSIDES, HAVE DIFFERENT KIND OF RECOGNITION HEADS AND BY THE SAME TOKEN WE CAN ALSO CHANGE THE INTRACELLULAR PART AND PUT ON DIFFERENT TRAN DESCRIPTION ALDO MAIN THAT'S RECOGNIZE DIFFERENT PROMOTERS SO WE COULD HAVE A
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WHOLE SET, ORTHOGONAL TO EACH OTHER. THESE CAN ALL FUNCTION IN THE SAME CELL. THAT'S AN INCREDIBLY FLEXIBLE WAY TO CHANGE RELATIONSHIPS BETWEEN CELLS AND INDUCE NEW RESPONSES. VERY QUICKLY ABOUT SYNTHETIC ADHESION MOLECULES. THEY HAVE THIS, THEY HAVE TO BIND TO EACH OTHER TO FORM TISSUE, TO THE EXTRA CELLULAR MATRIX AND TO MIGRATE OR TRANSMIGRATE THROUGH THE ENDOTHELIUM AND IMPORTANT FOR T CELL SYNAPSE OR NEURONAL SYNAPSES. THERE ARE A LOT OF COMPLEX ADHESION MOL KEWELLS.
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WE COULD ENGINEER SYNTHETIC WE REPLACE THESE EXTRA CELLULAR DOMAINS WITH ANTIGEN INTERACTIONS OR GFP BUT DEPENDING ON WHAT KIND OF INTRACELLULAR DOMAIN WE PUT ON THE INSIDE AND HOW IT ENGAGES THE INTRACELLULAR, CYTOSKELETON WE COULD GET DIFFERENT STRENGTH AND MORPHOLOGICAL DEPEN DEN SY. WHAT IS SHOWN HERE IS AN EXAMPLE, IF WE TAKE GFP INTERACTION BUT HAVE IT TIED TO THE INTRACELLULAR, WE GET SOMETHING THAT REPLICATES BROAD AND TIGHT INTERFACE WITH CADHERIN.
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THIS ALLOWS US TO TUNE STRENGTH AND SYNAPSE PROPERTIES FOR SORTING AND ASSEMBLY. I WILL SHOW YOU MOVIES THESE ARE THINGS WE ENGINEERED THE BLUE CELLS WITH DIFFERENT ADHESIONS THEY COULD EITHER FORM A NODE OR FORM THIS SHELL AROUND THE CLUSTER SO WE COULD START PROGRAMMING THOSE BEHAVIORS AND WE HAVE USED THAT IN A LOT OF DIFFERENT WAYS. LET ME GO TO THE MAIN PART OF THE TALK HOW WE CAN APPLY THIS TO NEXT GEN CELL THERAPIES. EVERYONE KNOWS ABOUT CAR T THERAPIES ONE OF THE FIRST CELL
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THERAPIES FDA A PROVED, THANKS TO PIONEERING WORK BY JIM -- WE KNOW WE CAN MAKE THESE ARTIFICIAL RECEPTORS THAT RECOGNIZE ANTIGENS ON CANCER AND KILL THEM AND THIS HAS BEEN REMARKABLE ADVANCE FOR HEMOPOETIC CANCERS. WE HAVEN'T HAD AS MUCH SUCCESS IN TARGETING SOLID TUMORS WITH CAR T'S. BECAUSE IT'S NOT REALLY THAT SIMPLE AS RECOGNIZING THE CANCER CELLS. RECOGNIZING CD 19 KILLS ALL T CELLS IS TOLERABLE AND THAT'S HOW IT CAN WORK. WE NEED A CELL THAT CAN
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RECOGNIZE THE ANTIGENS ON THE CANCER. BUT EQUALLY IMPORTANT, MANY ANTIGENS ESPECIALLY FOR SOLID TUMOR ARE ALSO EXPRESSED ON NORMAL TISSUE. WE HAVE TO FIND WAYS TO AVOID TOXIC CROSS REACTION. IN ADDITION MANY TUMOR SPECIFIC ARE ALSO HETEROGENEOUS. CERTAIN CELLS CAN ESCAPE. IF YOU GET TOO SPECIFIC YOU HAVE PROBLEM OF MORE CELLS MIGHT ESCAPE. ALSO SOLID TUMORS TEND TO BE SUPPRESSIVE, SO YOU HAVE TO OVERCOME THE SUPPRESSIVE TUMOR MICRO ENVIRONMENT. THERE ARE MULTIPLE CHALLENGE
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THAT'S HAVE TO BE SOLVED IN CONCERT TO GET THIS THERAPY. PART OF OUR GOAL IS DEPLOY SOME OF THE DIFFERENT MODULAR TOOLS WE HAVE TO IMPROVE EACH OF THESE DIFFERENT ISSUES AND TRY TO DO IT IN CONCERT. LET ME TELL YOU ABOUT ONE SOLUTION WE HAVE BEEN WORKING ON FOR QUITE SOME TIME. FIRST FOCUSED ON INCREASING RECOGNITION SPECIFICITY OF SAY SOLID TUMORS BUT HAS OTHER IMPLICATIONS ON SOME OTHER PROBLEMS. CAN WE PROGRAM FOR EXAMPLE A T CELL TO RECOGNIZE A MULTI-ANTIGEN SIGNATURE USING A
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TWO RECEPTOR CIRCUIT WHERE WE COMBINE SYN NOTCH AND A CAR INTO A KIR SET. KIR SET. -- CIRCUIT. IN NORMAL TISSUES COULD YOU GET THIS CROSS REACTION THAT COULD BE TOXIC. THIS IS WHAT I WAS TALKING ABOUT, WE COULD MAKE A CELL THAT HAS A SYN NOTCH RECEPTOR THAT WOULD RECOGNIZE ANTIGEN A THAT WOULD BE IN THE TUMOR. IT WOULD BE LINKED TO THE EXPRESSION OF A CAR GENE, SO THAT CAR IS NOT GOING TO BE EXPRESS BUD ONLY -- EXPRESSED. IF WE MAKE THIS TWO RECEPTOR
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CIRCUIT, BASICALLY IT'S OFF SO IT WOULD SPARE THAT HAD AND GENT B BUT TISSUE THAT HAS ANTIGEN A ON A CANCER CELL OR PART OF THE CANCER MICRO ENVIRONMENT IT WOULD TURN ON THE SYN NOTCH AND THIS WOULD TURN ON THE CAR, AND THE CAR WOULD BE ABLE TO INDUCE EXPRESSION OF THE CAR AND KILL THE CELLS THAT HAD ANTIGEN B. BUT ONLY IN THE CASE WHERE THE A AND B WERE TOGETHER LOCALLY IN SPACE, OKAY. AND WHAT IS SHOWN HERE IS AN ANIMATION OF WHAT WE HOPED WOULD HAPPEN.
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AS WE TAKE THESE SORT OF CAR SYN NOTCH T-CELLS AND PUT THEM IN A MOUSE WITH TWO TUMORS, ONE WITH THE KILLING ANTIGEN, THEY WOULDN'T BE ACTIVATED AND BE NEUTRAL BUT WITH THE PRIMARY ANTIGEN IN GREEN THEY WOULD GET TURNED ON AND KILL THAT IN THAT CASE ON THAT SIDE. WE SHOW WE TAKE MODEL TUMORS PUT IN ONE THAT HAS THE KILLING, KILLING AND PRIMING, WE SEE THIS AMAZING DIFFERENCE IN THE SAME ANIMAL WHERE THE SYN CAR T CELLS ONLY KILL THAT ONE SIDE. THIS SHOWS THESE ARE THE
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CONTROL T-CELLS AREN'T KILLED IF YOU PUT IN THE NORMAL CAR T CELLS IT WILL KILL BOTH TUMORS BUT THE SYN NOTCH CAN DETERMINE BETWEEN THE SINGLE AND DUAL ANTIGEN. SO WE HAVE NOW MOVED FROM THE SIMPLE MODEL ENGINEERED TUMORS TO SEVERAL DIFFERENT THINGS. WE AND OUR COLLEAGUES HAVE WORKED ON DEPLOYING THIS IN
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GLIOBLASTOMA. THEY ONLY KILL WHERE THEY ARE IN PROXIMITY. CAR EXPRESSION SHUTS OFF ONCE THE CELLS LEAVE THE REGION OF PRIMING WITH A HALF-LIFE OF A FEW HOURS. SO THEY DON'T HAVE TIME TO MOUNT THIS MULTI-DAY RESPONSE YOU NEED TO CLEAR A TUMOR. IN ADDITION, IT'S IMPORTANT TO NOTE THAT THE PRIMING ANTIGEN IS NOW ONLY USED FOR RECOGNITION AND NOT FOR KILLING. SO IT'S REALLY JUST AN INFORMATIONAL MOLECULE THAT LOCALIZES WHERE THE KILLING WILL TAKE PLACE. IT DOESN'T NEED TO BE HOMOGENOUS.
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IT CREATES BLAST RADIUS THE CELLS PRIMED CAN NOW KILL ANY CELL THAT HAS THE KILLING ANTIGEN OVER A BLAST RADIUS. WE CAN HARNESS NEOANTIGENS THAT ARE SPECIFIC BUT HOMOGENOUS. THE KILLING ANTIGEN NEED NOT BE PERFECTLY SPECIFIC AS LONG AS DUAL ANTIGEN COMBINATION IS ONLY FOUND ON THE TUMOR. THAT'S IMPORTANT, IF WE PRIME AT A VERY LOCALIZED WAY WE COULD START KILLING BASED ON BROADER SET OF ANTIGENS INCLUDING ONE THAT'S MIGHT HAVE CROSS REACTION IN OTHER TISSUES, AS LONG AS THOSE DON'T
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OCCUR WITH THE PRIMING ANTIGEN. AND THEN IMPORTANTLY, CONDITIONAL INDUCTION OF THE CAR, GREATLY DECREASES THE DEGREE OF CAR T CELL EXHAUSTION. MANY CIRCUIT ARE MUCH BETTER CLEARING THE TUMORS IN THE CIRCUIT CONTEXT RATHER THAN CONSTITUENT OF CAR. ALSO IMPORTANTLY, I WON'T TALK ABOUT THIS BUT VERY IMPORTANT PART, SYN NOTCH COULD BE USED TO INDUCE INFLAMMATORY PAYLOADS WE HAVE DONE THIS WITH IL2 AND HAS POWERFUL EFFECT OVERCOMING THE MICRO ENVIRONMENT IN
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PANCREATIC AND MELANOMA TUMORS AND MORE EFFECTIVE THAN ARMORED FORMS OF CAR THAT ARE CONSTITUTIVE. I WANT TO TALK B WE HAVE DONE A LOT OF THIS WORK IN COLLABORATION WITH MY COLLEAGUE IN NEUROSURGERY AN EXPERT IN GLIOBLASTOMA, THERE ARE TWO WE HAVE BEEN DEVELOPING, ONE E-SYNC. A CIRCUIT THAT PRIMES WITH A TUMOR NEOANTIGEN, EGFRVIII. SPLICE VARIANT PART OF THE EXTRA CELLULAR DOMAIN. THIS IS TUMOR SPECIFIC. BUT WHEN THEY DID A TRIAL USING A CAR OF RECOGNIZING IT, IT
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WENT TO THE TUMOR AND KILLED THE EGFRVIII CELLS BUT IT ESCAPED AND SURVIVED WITH EDITING OF THAT ANTIGEN. SO IT'S VERY POWERFUL BUT WE WANT TO USE IT FOR KILLING IN A SELECTIVE WAY BUT INFORMATION PRIMING ANTIGEN, WE HAVE USED IT AS A SYN NOTCH RECEPTOR TO PRIME EXPRESSION OF A TWO-HEADED CAR. THESE ARE MORE HOMOGENOUS BUT IMPERFECTLY SPECIFIC BUT BY COMBINING IN THIS PARTICULAR WAY, ONE GIVES MORE COMPLETE KILLING AND ONE GIVES MORE LOCALIZED SPECIFICITY IT WORKS QUITE WELL.
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THIS HAS BEEN PUBLISHED. WE HAVE A 90-YEAR APPROVAL AND IN PHASE TWO OF CLINICAL TRIAL, WE ARE EXCITED ABOUT THAT. THIS OTHER ONE B-SYNC PRIME ONCOA NORMAL BRAIN-SPECIFIC ANTIGEN. BCAN ONLY FOUND IN THE BRAIN. WE COULD MAKE A SYN NOTCH THAT RECOGNIZE THAT'S MOLECULE IN THE BRAIN AND INDUCES THIS TWO-HEADED CAR, THE SAME ONE. AND THESE ANTIGENS ARE NOT EXPRESSED IN THE NORMAL BRAIN. IT ENDS UP BEING REALLY VERY SPECIFIC AND WE ARE VERY EXCITED ABOUT IT AND I WILL TELL YOU ABOUT THAT.
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ONE OF THE KEY CONCEPTS HERE IS THAT WE WOULD LIKE TO ACTUALLY BE ABLE TO RESTRICT A THERAPEUTIC, NOT TO JUST RECOGNIZE IT'S MOLECULAR TARGET BUT RESTRICT ANATOMICALLY IN A TISSUE WHERE IT ACTS. IT'S HARD TO DO THAT WITH A MOLECULAR DRUG. OFTEN BIND WITH MULTIPLE TISSUES BECAUSE OF THE SAME OR RELATED TARGETS ARE IN DIFFERENT TISSUES. LET'S SAY WE WANTED TO HAVE THIS ATTACK SOMETHING IN THE BRAIN. THEN ONE IT MIGHT HAVE TROUBLE GETTING INTO THE BRAIN BUT ALSO
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CROSS REACT WITH OTHER CITES. -- SITES. IF WE COULD INSTRUCT THE DRUG TO ONLY GO TO THE BRAIN IT WOULD SOLVE A LOT OF PROBLEMS, IT MIGHT BE MORE EFFECTIVE IN THE BRAIN WHERE THE DISEASE IS. BUT ALSO WE WOULD DROP OUT ALL THE CROSS REACTION THAT OCCURRED OUTSIDE THAT TISSUE. OF COURSE THIS IS HOW A LOT OF BIOLOGY WORKS. THINGS GO AND OCCUR IN THE SPECIFIC TISSUES AND THAT'S WHY THE SAME MOLECULES COULD BE USED IN DIFFERENT CONTEXT. HOW MIGHT WE DESIGN A CELL TO
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RESTRICT ITS ACTIVITY TO A SPECIFIC TISSUE LIKE THE BRAIN? WE HAVE TRIED TO IDENTIFY BRAIN OR CNS SPECIFIC EXTRA CELLULAR ANTIGEN AND DESIGN SYN NOTCH TO RECOGNIZE THESE, AND USE TO INDUCE EXPRESSION OF A THERAPEUTIC PAYLOAD. IN THE CASE OF A CANCER LIKE GLIOBLASTOMA WE COULD INDUCE EXPRESSION OF A CAR, ONLY KILL WITHIN THOSE ANTIGENS IN THE BRAIN CONTEXT. BUT AS I WILL TELL YOU LATER WE ARE ALSO INTERESTED IN EXPRESSING OTHER PAYLOADS FOR NEUROINFLAMMATION COULD BE
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EXPRESSED SUPPRESSIVE PAYLOADS. SO WE DID WITH MILOS SIMIC LAB RESEARCH, THERE ARE CERTAIN MOLECULES IN THE BRAIN THAT ARE EXTRA CELLULAR THAT ARE QUITE SPECIFIC THAT INCLUDES CERTAIN SURFACE MOLECULES ON NEURONS OR
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ON OLIGO DEN SITES. EXTRA CELLULAR NETWORK, IT HAS HYLAURONIC, BUT THEN BCAN. WE RAISE ANTIBODIES AGAINST THIS AND MADE ELEVATED BCAN SYN NOTCH RECEPTORS.
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AS IT INTERACTS WITH THE P REGION AROUND THE ASTROCYTE IN GREEN. HOW DO WE APPLY THIS TO SOMETHING LIKE GLIOBLASTOMA, I WANT TO REVIEW, THE ANTIGENS WE CHOOSE TO USE FOR KILLING, WELL KNOWN GLIOBLASTOMA. BOTH OF THESE ARE WELL EXPRESSED ON MOST GLIOBLASTOMAS. BUT ONE OF THE PROBLEMS IS THAT THESE ARE ALSO EXPRESSED IN OTHER NORMAL TISSUES BUT NOT IN THE BRAIN. NOT IN THE NORMAL BRAIN. SO WHAT WE DECIDED TO DO, FROM MILOS SIMIC LEADING THIS WORK, TO MAKE A SYN NOTCH AGAINST
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BCAN AND HAVE THAT SO IF WE DETECT THE BRAIN, THAT WILL TURN ON THE CAR THAT WILL KILL THESE GLIOMA ASSOCIATED ANTIGENS EVEN THOUGH THEY AREN'T ABSOLUTELY SPECIFIC BUT THE ONLY ONES IN THE BRAIN, SO THE IDEA IS THE INTERSECTION OF THESE TWO VENN DIAGRAMS WOULD BE THE GLIOBLASTOMA. SO THEIR LAB MADE THESE CELLS, TESTED IN VITRO AND NOW INVIVO WITH A GBM TEW MORE, PBK MODEL IMPLANTED IN THE BRAIN. THIS ROBUSTLY CLEARED THESE TUMORS FROM THESE, INCREASED SURVIVAL, NEARLY ALL THE MICE
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SURVIVED. THIS SOMEONE OF THE STRONGEST RESPONSES WE HAVE SEEN OF ALL OUR GLIOBLASTOMA THERAPIES. IT WAS ACTING VERY EFFECTIVELY AND WE ARE GETTING A LOT OF PRIMING. WE TAKE THE TISSUE EARLY MID TREATMENT AND WE LOOK AT THE CD3 T CELLS THERE'S ONLY SIGNIFICANT ACTIVATION OF THE SYN NOTCH IN THE BRAIN SAMPLE. SO WE ARE VERY EXCITED ABOUT THAT, SEEMS TO BE VERY STRONG AND BRAIN SPECIFIC INDUCTION OF THE SYN NOTCH. ONE OF THE THINGS WE ARE MOST EXCITED IS THIS COULD BE A PAN
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GBM THERAPY, IT'S NOT THAT WE HAVE TO FIND PATIENTS THAT HAVE THAT SPECIFICKING THIS SHOULD WORK FOR ALL PATIENTS, THAT'S A BIG DEAL THINKING ABOUT HOW TO SCREEN AND HOW HARD IT IS FOR FINDING THE RIGHT PATIENTS. WHAT ABOUT OUTSIDE OF THE BRAIN? WHAT WE DID IS TOOK THE SAME TUMOR CELLS AND PLANTED ONE IN THE BRAIN AND ONE IN THE FLANK. NECESSITY -- THEY BOTH HAVE THE KILLING ANTIGEN BUT ONLY THE ONE IN THE BRAIN. IN THE SAME MICE, BRAIN TUMORS ARE CLEARED BUT THE FLANK
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IMPLANTED TUMORS ARE NOT CLEARED IN THE SAME ANIMALS. IT REALLY INDICATES THAT THIS IS BRAIN RESTRICTED ACTIVITY. OKAY, SO WE NOW ACTUALLY ARE REALLY EXCITED THAT WE HAVE THIS KIND OF GPS SYSTEM THAT CELLS COULD KNOW THEY ARE IN THE BRAIN. WE HAVE INTERESTED RESULTS ABOUT THINGS IN LUNG AND PANCREAS ALSO. BUT WHAT WE REALIZE THIS COULD BE USED FOR OTHER DISEASES TO DELIVER TO THE BRAIN. THERE'S BEEN A LOT OF WORK IN THE PAST PEOPLE TRYING TO FIND DIFFERENT CYTOKINES OR FACTOR
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THAT'S COULD HAVE DIFFERENT IMPACT ON NEUROINFLAMMATION, EA MODELS, SOME WORK WAS SHOWN TEN YEARS AGO, SOMETHING LIKE IL-10, SUPPRESSIVE CYTOKINE WHEN EXPRESSED IN THE BRAIN, WHEN YOU INJECT AAV IN THE BRAIN, THE HIGH LEVEL EXPRESSION CAN BLOCK EAE BUT SYSTEMICALLY INJECTED IL 10 IS INEFFECTIVE, EITHER NOT ABLE TO GET TO THE BRAIN OR HIGH ENOUGH LEVEL. MILOS WORKING WITH COLLEAGUES, INCLUDING SCOTT ZANVILLE IN THE NEUROLOGY DEPARTMENT ASKED CAN WE MAKE A T-CELL WITH SYN NOTCH
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DELIVER IL 10 AND HOW WOULD THIS WORK WITH MULTIPLE SCLEROSIS. LET ME TELL YOU ABOUT THE ANIMALS, MOG, ANTIGEN FROM MYELIN SHEATH CELLS. TH 17 AND ADOPTIVELY TRANSFER THAT INTO A MOUSE AND THAT MOUSE WILL DEVELOP NEURODEGENERATION AND NEUROINFLAMMATION. IN THIS CASE WE WOULD ASK COULD WE TRANSFER THE SYN NOTCH, THERAPEUTIC CELLS INTO THOSE CELLS AND WOULD THOSE HAVE EFFECTS ON THE EAE? WE LOOK AT DIFFERENT EFFECTS, NEUROLOGICAL SCORES, PARALYSIS, ET CETERA, WHAT WE SEE IN FACT
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WHEN WE GIVE THESE CELLS WE COULD SEE SIGNIFICANT AMELIORATION. BY DAY 12 THEY ADOPTED EAE BUT THE MICE TREATED WITH THOSE SUPPRESSORS CELLS MAINTAINED GOOD MOBILITY. WE ARE ABLE TO SEE AN EFFECT. WE ARE EXCITED THESE ARE EARLY DAYS BUT IDEA WE CAN TARGET CELLS TO TISSUE. WE ENGINEERED T-CELLS THAT USE MULTI-RECEPTOR CIRCUIT, MULTIPLE COMPONENTS LINKED TOGETHER. TO MAKE PRECISE DISEASE-SPECIFIC DECISIONS AS AN EXAMPLE IN THE CASE OF GLIOBLASTOMA WE ARE ABLE TO
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ENGINEER PRECISE BRAIN CANCER CELL THERAPIES BY PROGRAM CELLS THAT ONE CAN SENSE THEY ARE IN THE BRAIN AND INDUCE EXPRESSION OF A CAR. THIS IS AN EXAMPLE OF A POWERFUL EXPRESSION OF ANATOMICAL AND MOLECULAR SPECIFICITIES THAT REALLY ONLY A CELL CAN DO EFFECTIVELY. WE ARE EXCITED THAT WE HAVE DEVELOPED A WAY TO HAVE BRAIN SENSING CELLS THAT COULD BE USED IN A DISEASE-AGNOSTIC MANNER TO DELIVER EITHER IMMUNE SUPPRESSIVE BUT ALSO REGENERATIVE PIE LOADS OR ONES FOR NEODEGENERATION.
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WE ARE TRYING TO IMPROVE CELL MIGRATION, TRANSMIGRATION ACROSS THE BBB BUT ALSO IT'S POSSIBLE WE COULD TARGET OTHER TISSUES IN AN ANALOGOUS KIND OF GPS, CELL GPS MECHANISM. I WANT TO REMIND EVERYONE WE ARE REALLY EXCITED WE ARE STARTING THIS EGRB3 WITH THIS TEAM AT UCSF AS WELL AS MID STREAM IN TERMS OF PREPARING OUR IND FOR THE BRAIN-PRIMED BCAN ONE, HOPEFULLY WE WILL BE ABLE TO REPORT ON THAT SOON. SO THE LAST PART OF MY TALK, I WANT TO TALK ABOUT TRYING TO
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APPLY THESE SAME FUNDAMENTAL TOOLS TO A DIFFERENT PROBLEMS FACED IN BIOLOGY. BUT ONE THAT STILL REQUIRES CELLS TO TALK TO EACH OTHER IN VERY PRECISE PROGRAMS IN THE CASE OF DEVELOPMENT. HOW CELLS INTERACT WITH ONE ANOTHER TO CHANGE AND ORGANIZE IN SPACE. THAT COMPLEX BEHAVIOR COULD BE PRE-PROGRAMMED INTO A SET OF DNA. SO THIS, I WILL SHOW YOU IN THIS MOVIE, ACTUALLY SOME SIMPLE CASES WE TOOK, GOT THEM TO ORGANIZE IN DIFFERENT THINGS. SPONTANEOUS WAY. SO WHY GENE DEVELOPMENT, WE ARE
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INTERESTED FROM A BOTTOM-UP VIEW. ARE THEY UTILIZED IN DIFFERENT DEVELOPMENTAL PROGRAMS AND HOW THEY ARE PUT TOGETHER TO CONSTRUCT THE DIFFERENT MORPHOLOGIES WE SEE IN METZOANS. FOR TRANSPLANT, MIGHT IT BE BETTER TO HAVE CERTAIN PROPERTIES UNIQUE TO THAT. THE FUNDAMENTAL QUESTION, PART OF WHAT WE ARE TRYING TO DO AT A MORE BASIC LEVEL IS, TO ASK, CAN WE USE THE SAME SORT OF TOOLKIT WE HAVE BEEN USING FOR ENGINEERING THE T-CELLS TO ALSO ENGINEER DEVELOPMENT? ARE THOSE REALLY THE
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FUNDAMENTAL RELATIONSHIPS THAT ARE POWERFUL FOR CREATING THESE NEW PROGRAMS? AND SO SEVERAL YEARS AGO, TODA AND MARSUIT WHEN POST DOCS THEY ASKED CAN WE TAKE CELL LINES LIKE IL 19 OR LESS THAT DON'T SELF ORGANIZATION AND PUT IN PROGRAMS THAT WOULD. WHAT IS THE BASIC LOGIC OF DEVELOPMENTAL PROGRAMS, ONE HYPOTHESIS, MOST PATHWAYS IN DEVELOPMENT HAVE CELL THAT'S TALK TO ONE ANOTHER AND THOSE INDUCE SELECTIVE CHANGES IN MORPH FOLG. SO CAN WE USE SYN NOTCH TO ENGINEER TO RECOGNIZE EACH
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OTHER BUT LINK THOSE TO THE INDUCTION OF VARIOUS MORPHOLOGICAL, THINGS LIKE ADHESION, COULD WE INDUCE ADHESION MOLECULES THAT CHANGE HOW THE CELLS SORT, AS STANDARD FOR DIFFERENTIATION, COULD WE INTRODUCE FLUORESCENT PROTEINS BUT ALSO INTRODUCE NEW SIGNALS LIKE SYN NOTCH LIGANDS THAT WOULD CARRY ON THIS CONVERSATION FURTHER. SO THEY DID THAT, THIS JUST SHOWS SOME OF THE THINGS THEY WERE ABLE TO DO, THIS WAS A CELL POPULATION ABLE TO INTERACT WITH ONE ANOTHER AND
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YOU GET THIS DIFFERENTIATION INTO GREEN AND RED POPULATION, IN THIS CASE, THEY ORGANIZE IN THIS POLARIZED FASHION. THE NEXT EXAMPLE IS ONE WHERE WE HAVE SIMILAR SET OF CELLS WE PUT TOGETHER, THEY START INTERACTING, WHAT YOU WILL SEE IS FORMATION OF A GREEN-COLORED CORE AND THAT LEADS TO FORMATION OF AN INTERMEDIATE RED OR M-CHERRY INTERMEDIATE SHELL. JUST TO EXPLAIN HOW THIS WORKS, IN THIS LAST CASE THIS IS RECIPROCAL CONVERSATION BETWEEN TWO CELLS. TWO CELLS, A AND B SHOWN AT THE
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TOP. THE A CELL HAS A SIGNAL THAT WILL TURN ON A SYN NOTCH AND B WILL TURN ON GFP AND HIGH LEVEL OF KADHERON. TURNS OUT THAT WAS AN EXTRA CELLULAR, THAT TURNS ON M SHARING AND INTERMEDIATE LEVEL. THOSE CELLS TURN RED AND FORM THIS SHELL. THIS HAPPENS IN A VERY SPONTANEOUS AND STEREOTYPED WAY CONSISTENTLY. WE CAN CUT THEM AND THEY WILL STILL REGENERATE THAT STRUCTURE, ET CETERA. WE COULD GET SIMPLE SELF ORGANIZATION. THIS IS ANOTHER EXAMPLE OF A LOT OF DEVELOPMENT HAS LARGER
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SCALE INGREDIENTS THAT LEAD TO PATTERNING. TODA SHOWED HE COULD MAKE A SYN NOTCH, BY MAKING SECRETOR CELLS THAT WOULD PRODUCE A GFP IN THIS CASE, WHAT WE CALL A BODY, BOTH ANCHOR CELLS THAT WOULD TRAP THE GFP AND THEN COULD ALLOW IT TO BE DETECTED BY A TELL WITH AN ANTIGFP SYN NOTCH. SENDERS AND BODY OF CELLS, FOUR MILLILITER SO FAIRLY LARGE SCALE. WHAT WE SEE WE ARE ABLE TO GENERATE A GRADIENT OF RESPONSE THAT EMANATES FROM THAT SOURCE. AND IF WE NOW ELABORATE THAT
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WITH VARIOUS POSITIVE AND NEGATIVE FEEDBACK LOOPS THAT OCCUR IN THE BODY, THIS ARTIFICIAL BODY OF CELLS WE ARE ABLE TO GET THE SAME BODY TO RESPOND IN DIFFERENT WAYS TO THE GRADIENTS AND WE GET DIFFERENT KINDS OF PATTERNS LIKE SHOWN ON THE RIGHT SIDE. A FRENCH FLAG PATTERN, THE CLASSIC RESPONSES THAT HAVE BEEN THEORIZED IN DEVELOPMENTAL BIOLOGY. I WON'T TALK, MUCH OF THAT HAS BEEN PUBLISHED AND THERE'S A LOT OF INTERESTING STUFF THERE. BUT THAT'S KIND OF TAKING CELLS
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THAT DON'T DEVELOP AND TRY TO MAKE SIMPLE THINGS FROM SCRATCH. WHAT ABOUT TRYING TO GUIDE LIKE STEM CELLS. ONE THING WE HAVE BEEN INTERESTED IN MORE RECENTLY, ASKING IF WE CAN TAKE ES CELLS AND ENGINEER CELLS THAT WILL TALK TO THEM AND SPATIALLY DEFINED WAYS. THESE ARE WHAT WE ARE CALLING SYNTHETIC ORGANIZER CELLS THAT COULD PROVIDE MORPH GENIC INSTRUCTION. WE CAN GET PRETTY INTERESTING THINGS TO COME OUT OF THAT. YOU KNOW, THE IDEA IS WHEN WE LOOK AT DEVELOPMENT AND TRY TO
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DRIVE DIFFERENTIATION IN VITRO, MOST OF THE TIME WE ARE TAKING ES CELLS AND ADDING MORPHOGENS, MAYBE CHANGING THE TIME AND DOSE BUT DOING THIS IN THIS ISOTROPIC WAY. VERY DIFFERENT FROM WHAT HAPPENS IN VIVO, THERE ARE CELLS IN SPECIFIC LOCATIONS PRODUCING SPECIFIC MORPHOGENS. SOME EXCITING WORK ON EMBRYOS WORK FROM VARIOUS LAB THAT'S SHOW EXTRA EMBRYONIC CELLS CAN SELF ORGANIZE AROUND ES CELLS THAT LEAD TO QUITE ADVANCED IN VITRO EMBRYO GENESIS. CAN WE TRY TO RECAPITULATE THAT.
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REENGINEER SYNTHETIC CELL THAT'S WOULD ORGANIZE IN SPACE AROUND THE ES CELLS AND BE NOW THE SOURCES OF THESE MORPHOGENS. WE NEED TO PROGRAM SPATIAL CELL ASSEMBLY. NATURAL ADHESION AND ENGINEERED ONES WE COULD PROGRAM WITH SYNTHETIC ADHESION MOLECULES. AS I SHOWED BEFORE THESE MOVIES ARE FIBROBLAST CELL LINES ENGINEERED, THE SAME ONES BUT ENGINEERED IN DIFFERENT WAYS TO ORGANIZE AROUND THESE RED ES CELLS IN EITHER FORMING A NODE OR A SHELL. WHAT WE CAN DO WITH THESE IS
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NOW COMBINE IN DIFFERENT WAYS AND GET DIFFERENT ARCHITECTURES, SINGLE NODE, SHELLS COMBINED TO GET DIFFERENT PATTERNS AND TRY TO EXPRESS MORPHOGENS FROM THESE ANISOTROPIC WAYS. THE FIRST THING, THIS IS WORK FROM TOSHI, POST DOC IN THE LAB, IS TO ASK CAN WE DELIVER THIS KEY EARLY EMBRYONIC MORPHOGEN WITH 3A FROM SPECIFIC ORGANIZER CELLS AND WHAT WE FIND IS WE CAN GENERATE VERY TUNABLE AND DIRECTIONAL GRADIENTS. THESE ARE ES CELLS WITH PTCF MCHERRY, THAT'S WHY IT WILL TURN ON.
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IF WE GIVE A WNT AGONIST, IT TURNS ON, IN THE MEDIA IT TURNS EVERYTHING ON IN THIS ISOTROPIC SPHERICAL WAY. IF WE PROVIDE WNT EXPRESS FROM SHELL, WE SEE SOMETHING VERY SIMILAR. BUT IF WE EXPRESS WNT FROM THIS NODE WE SEE POLARIZED ACTIVATION, GRADIENT THAT GOES ACROSS THAT AND ELON GAIGS OF THE BODY ALONG WHAT LOOKS LIKE A PRIMITIVE ANTERIOR, POS STEEROR AXIS. IF WE MAKE A SECOND NODE AND PUT IT 180° AWAY AND MAKE A WNT INHIBITOR DKK1 WE CAN GET STEEPER, ACROSS THE SPAN OF
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THAT BODY, IF YOU WILL. SO WHAT HAPPENS AS WE TUNE THESE GRADIENTS? WHAT HAPPENS TO THE MORPHOLOGICAL OUTPUTS? WHAT IS SHOWN HERE AFTER EIGHT DAYS SOME OF THE THINGS WE SEE. I WANT TO START WITH THE MIDDLE ONE. WHICH IS WHERE WE HAVE BOTH THE WNT AND DKK NODES, THE INHIBITOR, THIS LEADS TO STEEP GRADIENT WITH HIGH DYNAMIC RANGE ACROSS THE WHOLE BODY. WE SEE SPECIFIC TISSUES ARRANGED IN MORE OR LESS THE CORRECT ANTERIOR, POS STEEROR ARRANGEMENT. WE SEE THINGS LIKE OTX2 NEURAL
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MARKERS AROUND THE DKK1 NODE AND THEN VARIOUS MARKERS FOR EXAMPLE, THIS IS CARDIAC TISSUE, VERY LOCALIZED IN THE MIDDLE. WE THINK THIS IS THE ANTERIOR AND POS STEEROR IN THIS DUAL NODE STRUCTURE. INTERESTINGLY THOUGH, IF WE USE SINGLE NODE WITH DKK1 WE GET ALMOST ALL NEURAL FATES. WE SEE POSTERIOR AND THE CARDIAC TISSUE AT THE MOST ANTERIOR THERE. AND SO WHAT WE SEE, WE CAN SEE SPECIFIC LINEAGES ARISING AND ORGANIZED IN THE APPROPRIATE ANTERIOR POSTERIOR TOPOLOGY BUT
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THE RANGE AND SHAPE AND STEEPNESS DETERMINE THE SPECTRUM OF LINEAGES WE OBSERVED IN THE RESULTING EMBRYOID. DUAL HEAD WITH THE MOST RANGE OF FATES. ONE THING I HOPE YOU CAN SEE IN THE MOVIE, THE BEATING CARDIAC TISSUE, IT'S LOCALIZED. THESE ARE SELF ORGANIZING ORGANIZED SUB TISSUES IN THIS ANTERIOR, POSTERIOR AXIS. IF WE LOOK MORE CAREFULLY IN THE CASE IF WE HAVE SINGLE WNT NODE, THAT HAPPENS THERE, THIS GRADIENT IS LESS STEEP AND YOU DON'T GET ALL THE TISSUES BUT
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IT'S SHALLOWER, WE FIND WE ACTUALLY GET INTERESTING MORPHOLOGY SUGGESTING HIGHER RESOLUTION SPATIAL OR MORPHOLOGICAL INFORMATION. ONE THING WE SEE PROBABLY ABOUT 15% OF THOSE EMBRYOIDS. WE SEE A CAVITY, AND HEART BY IMAGING HAS A REGULAR BEAT, WHICH IS QUITE UNEXPECTED AND REALLY DIFFICULT TO SEE IN A SPONTANEOUS SELF ORGANIZING WAY, THIS KIND OF STRUCTURE. MOST OTHER EFFORTS AT RECONSTRUCTING HEARTS HAVE INVOLVED TAKING MULTIPLE LINEAGES AND COMBINING THEM. NOT ONLY TO WE SEE THIS
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HEART-LIKE CAVITY. IT HAS MARKERS LIKE CARDIAC TRA CONE IN, OR TX2K5 LOCALIZED AROUND THE REGION, BUT WE SEE BY STAINING FOR ENDOTHELIAL MARKERS, IT LOOKS IT'S LINKED TO THE EXTENDED VASCULAR NETWORK MARKED IN GREEN. (NKX2.5) REALLY THIS IS AN AMAZING CASE OF SELF ORGANIZATION OF A MULTI-CELL TYPE HEART-LIKE STRUCTURE IN A SPONTANEOUS WAY. SO ONE OF THE THINGS WE ARE THINKING, THERE ARE KIND OF DIFFERENT WAYS OF LOOKING HOW TO DRIVE DEVELOPMENT IN VITRO OR RECONSTITUTING.
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SOME WORK IN THE PAST HAS BEEN TAKING ES CELLS AND USING COMBINATIONS OF PROTOCOLS OF DIFFERENTIATION FACTORS TO DRIVE DIFFERENTIATION OF SPECIFIC LINEAGES AND MAYBE YOU COMBINE THOSE IF YOU ARE INTERESTED IN TISSUES. ON THE SYNTHETIC EMBRYO SIDE USING FAIRLY NATURAL CELLS, CAN YOU PUSH ALL THE LINEAGES AND GET SELF ORGANIZATION OF THE WHOLE BODY. WE THINK WITH SOME SYNTHETIC ORGANIZERS WE CAN TUNE SOMEWHERE BETWEEN THAT TO DRIVE CERTAIN REGIONS OR RANGES OF
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LINEAGES AND STILL GET SOME SELF ORGANIZATION OR DRIVE THINGS TO NON NATURAL NON NATIVE FATES IN SOME CONTROLLED WAYS. THIS IS EARLY DAYS BUT WE ARE VERY EXCITED ABOUT THAT. WHAT WE HAVE SHOWN WITH THE SYNTHETIC DEVELOPMENT WORK, WE CAN COMBINE COMPONENTS THAT CONTROL CELL-CELL SIGNALING, DIFFERENTIATING CELL ADHESION AND MORPHOLOGICAL PRODUCTION FOR EXTRA CELLULAR MORPHOLOGIES.
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THEY CAN PROVIDE PROGRAMABLE CELLS WITH INFORMATION RICH SPATIAL THAT CAN GENERATE DISTINCT TISSUE, ASSOCIATED VASCULAR NETWORK, WE CAN CUSTOM TUNE THE GRADIENT RANGE AND STEEPNESS TO ALTER THE RANGE AND RESOLUTION OF THE MORPHOLOGICAL OUTCOMES. AND THIS APPROACH, WE THINK COULD BE APPLIED TO MANY DIFFERENT TYPES OF MORPHOGENS AS PROGENITOR CELLS. COULD WE USE THIS APPROACH IN VIVO TO GENERATE OR RECONSTRUCT AROUND SITES OF INJURY TO DRIVE REGENERATION IN SITU.
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A LONG WAYS AWAY BUT REALLY EXCITING. TO CONCLUDE THE WHOLE TALK I HOPE I HAVE BEEN ABLE TO SHOW YOU LIVING CELLS ARE THESE AMAZING MACHINES UNLIKE OTHER SCALES LIKE MOLECULES HAVE THIS MUCH HIGHER LEVEL TO INTEGRATE INFORMATION OVER A HIERARCHY OF SCALES, ANATOMICAL OR MOLECULAR, AND THEY CAN SPATIALLY COORDINATE BY SENSING AND INTERACTING WITH THEIR ENVIRONMENT OR NEIGHBORING CELLS. IF WE TAKE ADVANTAGE OF THIS, THERE ARE MANY THINGS WE CAN DO AND BUILD TOOLS AND SENSORS
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THAT ALLOW US TO PRECISELY CONTROL AND CHANGE THE CELLULAR INTERACTIONS AND BUILD DIFFERENT CELLULAR PROGRAMS AND CIRCUIT, INCLUDING USING THESE TWO OPERATING, MANY DIFFERENT CELLS INCLUDING IMMUNE CELLS THAT COULD SEARCH FOR CANCER OR TARGET TISSUE FOR SUPPRESSING INFLAMMATION OR PUTTING IN CELL THAT'S COULD PROVIDE SPATIAL INSTRUCTION THAT COULD GUIDE DEVELOPMENT. AND JUST WANT TO LEAVE YOU WITH THE THOUGHT THAT UNDERSTANDING HOW TO MANIPULATE THE LOGIC WILL ANSWER I THINK MANY
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QUESTIONS IN BIOLOGY WE DON'T UNDERSTAND. WE KNOW A LOT ABOUT THE COMPONENTS OF BIOLOGY, WE KNOW LESS HOW THEY ARE PUT TOGETHER, THE LOGIC. POWERFUL WAYS TO GENERATE NEW THERAPEUTIC PLATFORM THAT'S COULD INTERFACE WITH COMPLEX DISEASES LIKE CANCER OR FIBROSIS OR AUTOIMMUNITY. WE CAN DIRECT THE CONVERSATIONS THAT HAPPEN IN THESE ECOSYSTEMS. WITH THAT LET ME THANK ALL THE PEOPLE IN MY LAB AND COLLABORATORS. I WANT TO FOCUS, MILO SIMIC THE FELLOW LEADING THAT, WATCHMAKER. SCOTT ZAMVIL.
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ALSO BIO INFORMATICS DONE WITH MY COLLEAGUE AT PRINCETON. THE SYNTHETIC ORGANIZE ERICCA ALSTEAD WORK DONE BY SOSHI YAMADA IN COLLABORATION WITH OPHIR KLEIN. SO THANK YOU VERY MUCH. [ APPLAUSE ] >> ANYONE WITH QUESTIONS PLEASE USE THE MICKS. WE HAD A QUESTION THAT CAME IN, IF YOU ARE READY FOR IT. >> OKAY. >> SO HOW DO BRAIN TARGETED CAR T CELLS PENETRATE THE BLOOD-BRAIN BARRIER? >> WE KNOW THERE IS ALWAYS, I MEAN T-CELLS CAN GET IN THE
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BRAIN, THEY END UP CAUSING M.S., THEY CAN GET IN EARLY GBM TRIALS THAT EGFR CAR THEY DID GET INTO THE BRAIN. IT'S PRETTY CLEAR IN DISEASES LIKE BRAIN TUMORS AND M.S., THERE ARE SOME ALSO SOME DAMAGE TO THE BBB THAT ALLOWS CELLS TO GET IN EARLIER, MORE EFFECTIVELY. SO IT'S UNCLEAR, OUR CELLS ARE GETTING IN THERE, WE DO SEE THEM, THEY ARE PROLIFERATING. SO THEY DO GET IN. TO SOME DEGREE, WE THINK IT'S AMPLIFIED IN THESE PARTICULAR DISEASE CASES BUT WE ARE WORKING ON TRYING TO IMPROVE
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WAYS TO GET THEM THROUGH THE BBB. >> VERY BRILLIANT, AS USUAL. MY QUESTION IS ABOUT THE SPECIFICITY OF YOUR GBM TARGETING T-CELLS. YOU SHOWED BACKGROUND INFORMATION IN THE SPLEEN. DO YOU UNDERSTAND IT, IS THERE ENOUGH LIGANDS IN THE SPLEEN? WHAT IS HAPPENING THERE? IS THERE LEAKINESS. >> YOU ARE SEEING THE GLASS HALF EMPTY. >> VERY CONCERNED ABOUT BACKGROUND. >> IT'S MUCH, MUCH LOWER. THERE'S A LITTLE BIT, I WOULD SAY, WE, I GUESS, WE HAVE DONE
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EXTENSIVE NON-BRAIN TUMOR KILLING AND WE DON'T SEE ANYTHING THERE. WE DON'T SEE PROLIFERATION OF CELLS THERE. WE THINK WE ARE BELOW THE THRESHOLD FOR ANY SORT OF EFFECTS. >> I GUESS MORE GENERAL QUESTION, MOVING FROM THE MOUSE MODEL TO PATIENT, DO YOU HAVE A WAY TO TEST THE DIFFERENT TISSUES TO SEE -- >> WELL, I MEAN, WE'VE GONE THROUGH THE R&D PROCESS ON THIS AND WE ARE USING MOUSE CROSS REACTIVE ANTIBODIES. WE ARE DOING TOX IN THE MOUSE. ALSO RECHECKING EVERYTHING
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AGAINST HUMAN TISSUE LIBRARIES AND SO FAR THAT'S ALL CHECKED OUT. YOU KNOW, THIS IS WHAT A DOSE ESCALATION IS FOR. I WOULD SAY, A LOT OF THESE, THE EGFR 3, THAT CAR HAS BEEN IN TRIALS AND THERE WASN'T A PROBLEM. SO YOU KNOW, COMBINATION OF THINGS. BUT THERE ARE UNKNOWNS. YES? >> WHEN YOU WERE SHOWING US THE EXPRESSION OF THE WNT, AND YOU SHOWED US THE GRADIENT WHEN YOU INDUCED THE NODES AND INHIBITORS WHAT WERE YOU USING TO DIRECT THE NODES AND INHIBITORS?
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>> TO THE SECOND NODE? >> YES. >> THE SECOND NODE HAS MORE -- ELEMENT TO IT AND WE ARE TRYING TO IMPROVE THAT. >> OKAY, AND THE ON THE CARDIAC EXPRESSING CELLS, WHAT WERE THEY, SO THEY INDUCED THE VESSEL EXPRESSION IN THE NEARBY CELLS. DID THOSE VESSEL, WHAT DID THOSE VESSEL EXPRESSING DO TO THE NEIGHBORING CELLS? >> WENDELL LIM: YEAH, WE ARE JUST KICK-STARTING THE EVENT. THIS IS ONE SIGNAL. AND CLEARLY, THERE'S A LOT OF WNT PNB4, A LOT OF THINGS THAT
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GO ON IN REGIONS THERE. SO WE ARE TRYING TO TRACK BY SCOPE AND OTHER THINGS WHAT ARE THE SEQUEL LA OF EVENT THAT'S HAPPEN. THIS IS THE TRICKY THING. ONE TRIGGERING THE NEXT THING AND SO FORTH. SO THAT'S ALL TO BE WORKED OUT. >> THANK YOU. >> GREAT TALK. I HAVE A QUESTION ABOUT YOUR BRAIN-SPECIFIC SYN NOTCH CAR. AND YOU ARE USING TANDEM CAR? AND I'M WONDERING IF THERE'S ANY DIFFERENCE WHEN YOU HAVE ONLY SINGLE TARGET VERSUS TWO TARGETS, ANY RATIONALE BETWEEN USING TANDEM CAR?
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>> WENDELL LIM: YEAH, THE RATIONALE BEHIND USING THE TANDEM CAR HAS TO DO WITH THE FACT THAT EVERY PATIENT IS DIFFERENT, BUT THE NUMBER OF PATIENTS THAT HAVE HIGH RELATIVELY HOMOGENOUS EXPRESSION. IF YOU ARE LOOKING AT TANDEM, THAT'S AN ORGATE, KILLING ACROSS BOTH CIRCLES, RIGHT? WE WANT TO MAKE THE DECISION SWITCH OF SPECIFICITY TO BE VERY SPECIFIC AND WE WANT THE KILLING TO BE BROADER. >> ALL RIGHT, THANKS. >> I WANT TO BUILD OFF THE QUESTION ON THE SYNTHETIC
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ORGANIZER CELLS IN THE ASSEMBLY. IT'S INCREDIBLE YOU COULD SEE THE DEVELOPMENT OF THE CARDIAC TISSUE, BUT COULD YOU COMBINE THAT WITH YOUR SYN NOTCH TO FURTHER DEVELOP -- >> WENDELL LIM: THAT WAS REALLY OUR INTENT AT FIRST. THEN WE REALIZED WE HAVE TO DO FIRST THINGS FIRST. EVEN JUST LOCALIZING THEM WOULD BE VERY IMPORTANT. BUT YES, WE HAVE NOW THE ABILITY TO KILL NODES AND REPLACE THEM WITH NEW NODES AND WHAT WOULD BE REALLY POWERFUL, WHEN YOU TRY TO STRING TOGETHER
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THESE DIFFERENT EVENTS, AS IN DEVELOPMENT, IS TO BE ABLE TO NOW HAVE AN ORGANIZER THAT WOULD DETECT A MARKER OF THE NEXT STAGE OF DEVELOPMENT IN A LOCALIZED WAY. WE ARE TOTALLY INTO THAT. >> THANK YOU. >> HI, PROFESSOR, REALLY FANTASTIC WORK. I HAVE A KIND OF NAIVE QUESTION. THE OTHER LAB GOING TO USE SYN NOTCH FOR CUSTOMIZED INVIVO EXPERIENCE. HOW COULD WE UTILIZE THAT? ARE THOSE SYN NOTCH BUNCH OF PLASMA WITH MULTI-CODING OR ARE THERE COMPANIES WE CAN WORK
1:00:07
TOGETHER OR PAY FOR SERVICE? >> WENDELL LIM: THAT'S A GREAT QUESTION. I WOULD SAY THERE ARE NOW SEVERAL VERSIONS OF SYN NOTCH AND WHEN WE TAKE A NEW ANTIBODY, AN ANTIGEN THERE'S SOME DEGREE OF OPTIMIZATION WE NEED TO DO. IT ALSO DEPENDS ON SOME VERSIONS PRODUCE MORE PAYLOAD BUT MORE LEAKAGE, SOME ARE TIGHTER BUT HAVE LESS AND ALSO CELL-TYPE SPECIFIC THINGS. WE ARE TRYING TO COME UP, TO MAKE OUR LIVES EASIER BUT EVERYONE'S LIFE EASIER IS COME UP WITH A SMALL LIBRARY OF
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DIFFERENT TEMPLATES WHERE ONE COULD POP IN YOUR ANTIBODY, SCSV OF INTEREST, AND GUIDE THOSE INTO THINKING WHAT THE APPLICATION IS AND WHAT BEHAVIORS WOULD BE THE BEST. >> SO IF THIS IS LIKE A MINI LIBRARY AVAILABLE -- >> WENDELL LIM: NOT YET. A LOT OF THESE THINGS ARE AVAILABLE ON ADD GENE, BUT WE ARE CHANGING HEIGHTS OF EXTRA CELLULAR DOMAIN AND DIFFERENT SCAFFOLDS FOR THE RECEPTOR. WE ARE EXPANDING THAT FOR OUR OWN CASE. BUT WE WILL MAKE THAT AVAILABLE.
1:01:17
>> OKAY, THANK YOU. >> ONE MORE? >> THANK YOU FOR THE TALK. I HAVE A VERY GENERAL QUESTION. I'M THINKING WHETHER THERE ARE ANY EMERGING PHENOMENA IN THE WORK YOU PRESENTED. UNEXPECTED CELL STATE OR UNEXPECTED CELL FUNCTIONS? >> WENDELL LIM: IN THE T-CELLS OR DEVELOPMENTAL SYSTEMS? >> IN BOTH. >> WENDELL LIM: YEAH. WELL, THAT RAISES OBVIOUSLY A GREAT QUESTION OF ARE THERE POSSIBLE CELL STATES THAT YOU COULD DRIVE CELLS INTO, BE THEY
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IMMUNE CELLS OR STEM CELLS ALONG SOME TRAJECTORY. AND I SUSPECT THAT THERE ARE. I THINK IT'S A VERY FUNDAMENTAL QUESTION. YOU SEE A CERTAIN REALM OF SIMULATIONS DURING LIFE AND EVOLUTION. AND ARE THE CELL FATES OF THE T-CELLS AND STEM CELLS SO CANONIZED THEY STAY ON TRACK. THEY COULD GO OFF SOME. THERE ARE CELL THAT'S ARE HARD TO CLASSIFY. TO WHAT DEGREE, I DON'T KNOW. BUT THAT'S A VERY, SORT OF IMPORTANT AND FUNDAMENTAL QUESTION. >> THANK YOU. >> OKAY, THANK YOU VERY MUCH,
1:02:32
EVERYONE, FOR COMING TO THE LECTURE. AND THIS HAS BEEN A REALLY, YOU KNOW, LIVELY QUESTIONS AND ANSWERS. BUT YOU CAN CONTINUE CHATTING WITH THE SPEAKER IF YOU WANT. WE HAVE A RECEPTION JUST OUTSIDE THE AUDITORIUM. AND WITH THIS, THIS WRAPS UP THE LECTURE. THANK YOU. [ APPLAUSE ] WENDELL LIM: THANK YOU.
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