>> WE'RE ALL GOOD NOW.
I'M MATT WOLFE.
WELCOME TO CCG R.
A BIG WELCOME TO DAVE MOONEY
FROM HARVARD UNIVERSITY.
PLEASE TAKE IT AWAY.
[APPLAUSE]
>> THANK YOU VERY MUCH FOR THE
INVITATION AND OPPORTUNITY.
I'LL TALK ABOUT RESEARCH IN THE
CONTEXT OF USING BIOMATERIALS TO
IMPACT T CELL IMMUNITY, THIS IS
AN IMAGE FROM ONE OF THE
TECHNOLOGIES I'LL TALK ABOUT
TODAY, T CELLS COLORIZED IN
YELLOW ENGAGING WITH RED
MICROPARTICLES, ANTIGEN
REPRESENTING CELLS.
GOOD, THIS WORKS.
THE LAB HAS A VARIETY OF
COMMERCIAL ENGAGEMENTS, RELATIVE
TODAY THERE ARE A FEW, NOVARTIS
AND ATTIVARE AND LYELL, FUNDED
IN PART BY THE NCI ONCOLOGY
NETWORK, PART OF THE CANCER
MOONSHOT, PART OF THE NCI BILL
AND MELINDA GATES FOUNDATION AND
EARLY PARTS OF A LOT OF RESEARCH
FUNDED BY NIBIB.
SO, AS I MENTIONED, WE'RE
INTERESTED IN USING MATERIALS TO
CONTROL T CELL BIOLOGY, WE'RE
INTERESTED IN THIS OVER THE
ENTIRE LIFE CYCLE OF A T CELL IN
THE CONTEXT OF CANCER DEVELOPING
STRATEGIES MIMICKING KEY
FEATURES OF BONE MARROW TO TRY
TO DRIVE T CELL DIFFERENTIAL
FROM HEMATOPOIETIC STEM CELLS,
ENGINEERING HUMAN THYMIC TISSUE
TO CONTROL POSITIVE AND NEGATIVE
SELECTION, INTERESTED IN THE
IDEA OF -- THIS IS INTERESTING.
THE IMAGES ON THE BOTTOM ARE NOT
THE REAL IMAGES BUT THAT'S OKAY.
A LITTLE BIT MIXED UP.
THAT'S WHAT HAPPENS
TRANSITIONING FROM MAC AND PC,
USING BIOMATERIALS TO ADAPT AND
FINALLY IN THE CONTEXT OF
MODELING TUMORS.
I'LL TALK ABOUT THE LAST THREE
AND NOT THE FIRST TWO TODAY.
TO GET STARTED I'LL TALK ABOUT A
TECHNOLOGY WE'VE BEEN DEVELOPING
THAT WE HOPE WILL ENABLE US TO
HAVE AN IMPACT IN ADOPTIVE
T-CELL THERAPIES.
HERE AT THE NCI YOU'RE FAMILIAR
IN THE CONTEXT OF CANCER
THERAPY.
THE QUESTION WE BECAME
INTERESTED IN A FEW YEARS AGO
WHETHER WE COULD IMPROVE T CELL
MANUFACTURING AND IMPORTANTLY
CONTROL THE ACTIVITY OF THE
TRANSFERRED T CELLS.
THE APPROACH WE CAME AT THIS
FROM WAS TO TRY TO DEVELOP A
BETTER ANTIGEN-PRESENTING CELL
MIMIC.
YOU'RE FAMILIAR WITH USE OF
DYNABEADS TO EXPAND AND
MANUFACTURE T CELLS.
THESE PRESENT A LOT OF CUES IN
NON-PHYSIOLOGIC MANNER, BECAME
INTERESTED IN THE ANTIGEN
PRESENTING CELL MEMETIC SCAFFOLD
TO MIMIC NATURAL ANTIGEN
PRESENTATION AND OTHER CUES T
CELLS GET FROM APCs TO HAVE
BETTER CONTROL OVER T CELL
BIOLOGY.
THE SYSTEM WE DEVELOPED IS SHOWN
HERE.
THIS IS THE WORK OF ALEX CHUNG,
NOW AT LYELL, AND DAVID ZHANG
WHO FINISHED HIS Ph.D. IN THE
LAB AS WELL, STARTING WITH
MICROPARTICLES OF A SILICA THAT
DISSOLVES BY HYDROLYSIS, IT GOES
AWAY INSIDE OR OUTSIDE THE BODY,
THIS IS AN IMAGE OF WHAT
PARTICLES LOOK LIKE, LONG
ALIGNED NANOPORES DOWN THE
LENGTH OF THE PARTICLE PROVIDING
A REALLY USEFUL PLACE TO PUT
BIOLOGICAL CARGOES.
FOR EXAMPLE, WE CAN ABSORB
CYTOKINES KEY MEDIATORS OF
INTERACTION BETWEEN ANTIGEN
PRESENTING CELLS AND T CELLS IN
THE BODY.
HAD THESE RELEASED IN A
SUSTAINED AND LOCALIZED MANNER
TO INTERACTING T CELLS.
I'M JUST SHOWING DATA WITH A FEW
CUES HERE, IL-2, IL-21,
TGF-BETA, TYPICALLY ABOUT A WEEK
TO TWO WEEKS OF SUSTAINED
PRESENTATION OF CUES DIRECTLY
FROM THE MATERIALS.
NOW, TO ENGAGE THE T CELLS
DIRECTLY WITH T CELL
STIMULATION, CO-STIMULATION, WE
COAT THE MICROPARTICLES FIRST
WITH SUPPORTED LIPID BILAYER, AN
INTERFACE CONNECTED TO THAT
NORMAL THE T CELL IS SEEING IN
ANTIGEN PRESENTING CELL, INSTEAD
OF RIDGE IMPARTICLE IS A MIMIC
OF A CELL MEMBRANE, FRET STUDIES
SHOWING IT'S INDEED IT HAS
FLUIDITY AND THAT AFTER WE
BLEACH WE CAN GET RECOVERY
FOLLOWING BLEACHING.
WE CAN THEN CONJUGATE VARIOUS
TYPES OF SIGNALS WE WANT TO
PROVIDE DIRECT STIMULATION OR
CO-STIMULATION TO THE T CELLS,
AND KEY FEATURE HERE IS UNLIKE
NORMAL ANTIGEN PRESENTING CELL,
HERE WE CAN PRECISELY CONTROL
THE DENSITY AND NUMBER OF THESE
CUES WE PRESENT TO THE CELLS.
THIS WILL END UP BEING IMPORTANT
LATER, HERE WE'RE JUST SHOWING,
FOR EXAMPLE, WE CAN CONTROL
AMOUNT OF IgG, STIMULATORY
ANTIBODY PRESENTED FROM LIPID
MEMBRANES, COMBINE WITH T CELLS,
HERE A VIDEO SHOWING THE
MICROPARTICLES WE'VE INCLUDED
GREEN FLUORESCENT TAG IN LIPID
BILAYER, T CELLS IN RED.
NOW, THE FIRST THING I NEED TO
MENTION ABOUT THE SYSTEM IS THAT
WE CAN DO BOTH KIND OF
POLYCLONAL T CELL EXPANSION BY
PRESENTING ANTIBODIES THAT
ENGAGE TCR AS WELL AS
CO-STIMULATION OR WE CAN DO
STIMULATION IN ANTIGEN SPECIFIC
MANNER CONJUGATING, PRESENTING
THE RIGHT PEPTIDE ANTIGEN TO THE
CELLS AND I'LL DESCRIBES STUDIES
WITH BOTH OVER THE NEXT FEW
MINUTES.
THE FIRST STUDY ALEX DID WAS
SIMPLE EXPANSION STUDY TO USE
THESE TO MANUFACTURE LARGE
QUANTITIES OF T CELLS.
HE DID A SIMPLE COMPARISON OF
EXPANSION COMPARING APCMS TO
INDUSTRY STAND ART, DYNABEADS,
FOR THOSE NOT FAMILIAR THESE ARE
PLASTIC BEADS THAT HAVE
ANTIBODIES CONJUGATED TO THEIR
SURFACE.
WHAT ALEX DEMONSTRATED THESE
MORE METIC PARTICLES COULD LEAD
TO EXPANSION OF T CELLS QUITE
ROBUST T CELL EXPANSION,
ACTUALLY SIGNIFICANTLY
OUTPERFORMING DYNABEADS, THE
INDUSTRY STANDARD, IN TERMS OF
EXPANSION OF THE CELLS AND THIS
CAME WITHOUT THE COST OF
INDUCING EXHAUSTION, AS
INDICATED HERE BY PD-1 LAG3
STAINING ON T CELLS.
IT LOOKED LIKE A PROMISING
SYSTEM IN TERMS OF ENSURING WE
DID NOT LOSE FUNCTIONALITY WHEN
EXPANDING WITH THE SYSTEM, THE
CLASSIC EXPERIMENT HERE,
ADAPTIVE CELL THERAPY,
IMMUNOCOMPROMISED ANIMALS,
INTRODUCED HUMAN MODEL OF
CANCER, INTRODUCED CAR-T CELLS
THAT WE EXPANDED WITH DYNABEADS
OR APCMS SYSTEM AND TRANSFERRED
THE SAME NUMBER.
FROM THE GRAPHS ON THE BOTTOM IN
TERMS OF THE NORMALIZED TOTAL
FLUX WHICH GIVES AN INDICATION
OF THE TUMOR BURDEN IN THE
ANIMALS, THE SURVIVAL, UNTREATED
ANIMALS RAPIDLY SUCCUMBED, BOTH
THE DOSE OF DYNABEAD EXPANDED T
CELLS AND APCMS EXPANDED T CELLS
WERE FUNCTIONAL IN TERMS OF
BEING ABLE TO SLOW TUMOR
PROGRESSION AND APC-MS CELLS
WHICH WE HAD NOT MADE EFFORT,
OUTPERFORMED DYNABEADS, CELLS
MORE FAST, MORE CELLS MORE
RAPIDLY, WENT COMPROMISING TERMS
OF THEIR FUNCTION.
THIS WAS INTERESTING.
BUT WHAT WE WERE WANTING TO GET
AT IS COULD WE USE THIS KIND OF
SYSTEM, ENGINEERED SYSTEM TO
CONTROL THE FUNCTIONS OF T CELL
POPULATIONS THAT WE WANTED TO
DELIVER INTO THE BODY.
WHEN WE STARTED, WE'RE NOT SURE
AS A FIELD WHAT ARE OPTIMAL
CHARACTERISTICS OF A PRODUCT WE
WANT TO TREAT A PATIENT WITH.
THE APPROACH WAS A NON-BIASED
APPROACH, WHERE WITH THIS SYSTEM
WE CAN CONTROL FEATURES OF THE
MSRs, CONTROL THEIR PHYSICAL
DIMENSIONS, FOR EXAMPLE, WE CAN
CONTROL WHAT CUES, WHAT
PERACRINE ACCOUNT AING SIGNALS
WE PUT IN, CAN CONTROL
MECHANOTRANSDIGS BY CONTROLLING
LIPIDS THAT PRESENT THE CUES,
INSTEAD I'M GOING TO TALK ABOUT
SIMPLE EXPERIMENTS WITH
CONTROLLING THE AMOUNT OF
STIMULATION BY VARYING QUANTITY
OF T CELL LIGANDS.
THE APPROACH WAS VERY
NON-BIASED.
WE CREATED A LARGE LIBRARY OF
DIFFERENT APC-MS, VARY THE
AMOUNT OF STIMULATION, EXPAND T
CELLS AND LOOK AT A PANEL OF
MARKERS FROM T CELL
DIFFERENTIATION STATUS AND
PHENOTYPE.
AND AS MENTIONED WE VARIED T
CELL LIGANDS, WE WOULD COMPARE
ALL THESE DIFFERENT END RESULTS,
T CELL PRODUCTS, AND DO TYPICAL
PRINCIPAL COMPONENT APPROACH TO
LOOK AT FEATURES WE COULD
GENERATE IN T CELLS.
EACH ONE OF THE DOTS IS A
SEPARATE MANUFACTURING RUN, WITH
A SLIGHTLY DIFFERENT OR MAYBE
MAJORLY DIFFERENT VERSION OF
APC-MS SYSTEM, WE VARIED OVER A
BROAD RANGE, GENERATED LOTS OF T
CELL PRODUCTS WE CHARACTERIZED A
VARIETY OF WAYS, WHEN WE USED K
CLUSTER APPROACH TO BEND THEM
INTO GROUPS, YOU CAN SEE HERE WE
HAD ABOUT 8 GROUPS WE BEND THESE
INTO FOR SIMPLICITY, AND IF WE
STARTED TO LOOK AT PHENOTYPE OF
THESE DIFFERENT CLUSTERS WHAT WE
SEE IS THAT DEPENDING ON THE
SPECIFIC STIMULATION WE PROVIDE
WE CAN HAVE A DRAMATICALLY
DIFFERENT IMPACT ON THE
EXPANSION OF THE T CELLS.
IF WE LOOK AT THINGS LIKE CD4 TO
CD8 RATIO, BELIEVED TO BE AN
IMPORTANT METRIC, WE CAN VARY
THAT OVER A WIDE RANGE.
IF YOU LOOK AT EXHAUSTION, AGAIN
THESE CLUSTERS SHOW VERY
DIFFERENT PROPERTIES.
IF WE WANTED TO THINK ABOUT
CONTROLLING DIFFERENTIATION
STATUS BY LOOKING AT MARKERS
THAT ARE INDICATIVE OF MEMORY
STATUS OF THESE POPULATIONS,
AGAIN WE CAN VARY THIS QUITE
DRAMATICALLY SHOWN IN THE LOWER
RIGHT WITH DIFFERENT COLORS
THERE.
THE POINT OF THIS IS, THIS
PROVIDES A REALLY SIMPLE AND I
THINK FACILE APPROACH THAT
ALLOWS US TO TUNE THE TYPES OF T
CELL PRODUCTS THAT YOU GET FROM
A STARTING POPULATION.
WE CAN DO THIS GENERATE LARGE
NUMBERS OF CELLS WITH WIDE
VARIETY OF DIFFERENT TYPES OF
METRICS.
WE HAVE SOME IDEAS WHAT METRICS
MIGHT BE USEFUL FOR T CELL
PRODUCTS BUT AS THAT EVOLVES
THIS COULD BE A USEFUL SYSTEM TO
ALLOW US TO GENERATE DIFFERENT
KINDS OF PROPERTIES.
ONE OF THE INTERESTING FEATURES
IF WE COLOR CODE BASICALLY EACH
OF THESE INDIVIDUAL RUNS BY
ABSOLUTE AMOUNT OF STIMULATION
PROVIDING FROM CO-STIMULATORY
CUES, DARKER MEANS MORE, YOU CAN
SEE THE GENERAL TREND HERE WITH
THESE CELLS ALL DERIVED FROM
HEALTHY INDIVIDUALS, MORE
STIMULATION GENERALLY DOMINATED
PC 1.
NOW, WE BECAME INTERESTED IN
SAYING WHAT HAPPENS IF WE TAKE
CELLS FROM PATIENTS INSTEAD OF
HEALTHY INDIVIDUALS?
SO THIS WORK WAS DONE WITH KATHY
WU AND JERRY RITZ AT
DANA-FARBER, A LOT IN
COLLABORATION WITH NIKO, THE
POSTDOC IMAGE ON THE TOP, ON THE
LEFT, AND A Ph.D. STUDENT IN
THE LABORATORY, SO WE TAKE CELLS
FROM PATIENTS, WOULD LOOK TO
EXPAND THEM, AGAIN WITH INDUSTRY
STANDARD, DYNABEADS, OR APC-MS
SYSTEM AND CHARACTERIZE
FUNCTIONAL PROPERTIES WE GOT.
THIS GIVES A SENSE OF THE RANGE
OF STIMULATION WE PROVIDED.
FOR DIRECT COMPARISON, DYNABEAD
3:1 PROVIDES THE SAME CUES AS
APC-MS, AS A COMPARISON.
WHEN WE LOOK AT THE EXPANSION OF
THESE CELLS, YOU CAN SEE FIRST
OF ALL WE DON'T GET NEARLY THE
LEVEL OF EXPANSION WITH THE
PRIMARY CELLS FROM THE HEALTHY
DONORS, WE ALSO SEE A DIFFERENT
RESULT WHERE GENERALLY SPEAKING
WITH THE CELLS FROM HEALTHY
DONORS AS YOU INCREASE AMOUNT OF
STIMULATION YOU DRIVE MORE AND
MORE EXPANSION.
WITH THESE CELLS THAT'S NOT
ACCURATE.
AND INDEED IF YOU LOOK AT
DYNABEAD CONDITIONS AS WE GO
MORE DYNABEAD STIMULATION WE
DIMINISH THE EXPANSION WHILE
WITH APC-MS SYSTEM THERE'S
OPTIMAL, TOO LITTLE STIMULATION
DON'T GET A LOT, TOO MUCH WE
LOSE PROLIFERATION AS WELL.
NOW, IF YOU LOOK AT THE
FUNCTION OF THESE CELLS BY
LOOKING AT CYTOTOXIC ABILITY,
SIGNIFICANTLY DIFFERENT
CYTOTOXIC FUNCTIONALITY OF THESE
CELLS DEPENDING HOW WE
MANUFACTURE THEM.
ACTUALLY, THE ORANGE HERE, .1 IS
THE ONE THAT ALSO LED TO MAXIMAL
STIMULATION, OR MAXIMAL
EXPANSION.
SO IT'S NOT JUST WE'RE HAVING
INVERSE RELATIONSHIP BETWEEN
SUBSTANTIAL AND FUNCTION; WE CAN
ACTUALLY CREATE THESE SYSTEMS
WHERE WE CAN GET OPTIMAL
EXPANSION AND ALSO OPTIMIZE
FUNCTION SIMULTANEOUSLY.
NOW, TO DEMONSTRATE THIS HAD
RELEVANCE IN VIVO IN TERMS OF
TRUE FUNCTIONALITY, WE THEN WENT
AHEAD AND COMPARED CELLS
MANUFACTURED BASICALLY AT THREE
LEVELS OF STIMULATION, IN THE
SAME MODEL.
AGAIN USING CAR T CELL APPROACH.
AND WHAT YOU CAN APPRECIATE IS
EVEN THOUGH WE'RE DELIVERING THE
SAME NUMBER OF CELLS EXPANDED
WITH THE SAME MICROPARTICLE
SYSTEM, THAT DEPENDING ON THE
LEVEL OF STIMULATION WE PROVIDE
WE GET DRAMATIC CHANGES IN THE
FUNCTION.
AND YOU CAN APPRECIATE, FOR
EXAMPLE, THE CELLS STIMULATED AT
THE HIGH ARE MUCH LESS
FUNCTIONAL IN TERMS OF BEING
ABLE TO BASICALLY CLEAR THE
CANCER CELLS FROM THESE ANIMALS
AND HERE YOU JUST SEE SOME OF
THE SURVIVAL DATA AND SOME OF
THE BASICALLY TUMOR BURDEN LOAD.
SO WE CAN HAVE DRAMATIC IMPACT
IN CELL CULTURE AND THE ANIMAL
IN THESE TYPE OF SYSTEMS.
ONE OF THE CHALLENGES YOU CAN
APPRECIATE FROM THAT LAST SLIDE
IS EACH OF THOSE DOTS WAS AN
INDIVIDUAL PATIENT'S CELLS.
THERE'S GREAT VARIABILITY.
WE ALL APPRECIATE THAT WHEN WE
START GETTING TO REAL PATIENTS.
THERE'S A LOT OF VARIABILITY.
WE BEGAN TO WONDER HOW COULD WE
TRY TO DEAL WITH THIS AND MAYBE
MAKE A MORE HOMOGENOUS PRODUCT
AT THE END.
AND SO WE CAME UP WITH A CONCEPT
THAT MANY PEOPLE PURSUING
SIMILAR THINGS TODAY WHERE WE
COULD START WITH SOME
PHENOTYPING OF THE STARTING T
CELL POPULATION, SO WHAT ARE THE
CELLS.
IF WE SAID WHAT PRODUCT WE WANT
AT THE END, AND THEN WE
BASICALLY USE A.I. APPROACH, TO
FEED IN A LOT OF DATA, TO CREATE
A PRE- PREDICTIVE ALGORITHM HOW
DO WE STIMULATE CELLS FROM GIVEN
POPULATION TO GET TO ENDPOINT WE
WANT?
AND THIS SYSTEM AS I MENTIONED
IS EASY FOR US TO MANIPULATE.
WE TOOK THIS APPROACH, HAD
REASONABLE ACCURACY, 76% IN
TERMS OF BEING ABLE TO PREDICT
FUNCTIONAL CHARACTERISTICS OF
THE CELLS AT THE END, DEPENDING
ON THE INPUT APPROACH AND
MODULATING APC-MS STIMULATION.
SO WE THINK THIS CREATES A PATH
TO BE ABLE TO DEFINE AHEAD OF
TYPE WHAT CAR CAR T PRODUCTS OR
ADOPTIVE CELL WE WANT AND
PREDICT HOW WE NEED TO STIMULATE
WITH APC-MS TO GET TO CONSISTENT
PRODUCT IN THE END INDEPENDENTLY
OF INPUT PATIENT PROPERTIES,
THIS COULD BE USEFUL POTENTIALLY
FROM THAT PERSPECTIVE.
THE OTHER THING I WANT TO SPEND
A FEW MINUTES TALKING ABOUT IN
THE CONTEXT OF NOT POLYCLONAL T
CELL EXPANSION BUT
ANTIGEN-SPECIFIC T CELL
EXPANSION, MCI HAS BEEN A REAL
PIONEER AND HOME FOR TIL
STRATEGIES, WE'VE BEEN
INTERESTED IN ANTIGEN-SPECIFIC
T-CELL THERAPIES AS WELL.
AND ONE OF THE LIMITATIONS THERE
IS ABILITY TO REALLY EXPAND
THESE CELLS, PARTICULARLY WITH
THE APCs THAT COME FROM THE
SAME PARTICIPANT-CENTERED. --
PATIENT.
WE THOUGHT THAT MIGHT BE USEFUL
FROM THAT PERSPECTIVE.
FIRST WORK WAS WE LOOKED FOR
BASICALLY EXPANSION IN THIS CASE
MART 1 SPECIFIC T CELLS.
WHICH ARE BASICALLY COMMON T
CELL ANTIGEN, CONTEXT OF
MELANOMA, THAT EXISTS OFTENTIMES
IN OTHERWISE HEALTHY PATIENTS.
SO HERE WE'RE LOOKING AT ABILITY
TO EXPAND AND ENRICH MART1 CELLS
CELLS FROM PATIENT SAMPLES.
SIMP
DEPENDING ON AMOUNT OF
STIMULATION FROM ANTIGEN
PRESENTING MIMICS, .1, .5, 1%
RELATES TO HOW MUCH SUBSTITUTION
WE PUT IN ANTIBODIES ON THE
ARTICLES, YOU CAN SEE WE HAVE
DRAMATIC EFFECT ON EXPANSION AND
ENRICHMENT, AND WE GUT UP TO 10
TO THE 4th EXPANSION OF CELLS
OVER THE TIME COURSE OF A COUPLE
WEEKS WITH THE OPTIMIZED SYSTEM.
THIS WORK IS BEING DONE BY NIKO
IN CATHY'S LAB, AND A PEDIATRIC
HEMATOLOGIST/ONCOLOGIST AT
CHILDREN'S HOSPITAL, AND
DANA-FARBER DOING RESEARCH IN
THE LABORATORY.
NOW, ONE OF THE QUESTIONS IS HOW
DOES THIS FUNCTION RELATIVE TO
TYPICAL MONOCYTE DERIVED TCs
USED FOR EXPANSION OF THESE
CELLS?
WE'VE DONE DIRECT HEAD-TO-HEAD
COMPARISON, BOTH AGAIN IN TERMS
OF THEIR ABILITY TO ENRICH THE
POPULATION, AND ACTUALLY TO
EXPAND THE CELLS.
WHAT YOU CAN APPRECIATE FROM
BOTH THESE METRICS ON THE GRAPHS
ON THE LEFT THAT WE'RE ABLE TO
GET MUCH GREATER ENRICHMENT, AND
ABOUT, YOU KNOW, LOG, LOG AND A
HALF MORE EXPANSION OF THE MART1
SPECIFIC CELLS WITH THIS SYSTEM
THAN IS POSSIBLE WITH BASICALLY
THE NATURAL CELL ONE WOULD USE.
THERE'S GREAT VARIABILITY
DEPENDING ON SPECIFIC PATIENT
FROM FROM WHICH WE OBTAIN THE
CELLS.
WE'VE LOST A LOT OF PARTS ON THE
GRAFT BUT ON THE RIGHT YOU CAN
GET A SENSE OF THE DOT ON THE
LEFT, AND CIRCLE ON THE RIGHT,
IS CONNECTING INITIAL POPULATION
OF MART1 CELLS IN THAT SAMPLE,
THE BUBBLE ON THE RIGHT IS END
OF THE EXPANSION.
THERE'S A GREAT VARIABILITY
BETWEEN THE INDIVIDUAL PATIENT
SAMPLES, IN ALL CASES APC-MS LED
TO MUCH MORE DRAMATIC EXPANSION
OF THESE ANTIGEN-SPECIFIC CELLS
THAN DID THE MONOCYTE-DERIVED
DENDRITIC CELLS.
NOW, IN TERMS OF FUNCTION OF
THESE CELLS, WE'RE ALSO VERY
INTERESTED OBVIOUSLY NOT JUST IN
THE NUMBERS BUT ALSO THE
FUNCTION, AND WE'VE DEMONSTRATED
WE CAN GENERATE CELLS THAT LOOK
LIKE THEY HAVE A BETTER
PHENOTYPE IN TERMS OF ADOPTIVE
CELL THERAPY.
IN THIS WE'RE LOOKING AT GREEN
BASICALLY APC-MS-EXPANDED CELLS,
LIGHT BLUE MONOCYTE DERIVED, ON
THE LEFT A FEW MARKERS, IL7
RECEPTOR HLA-DR, PD-1, BETTER
PHENOACTIVIC CHARACTERISTICS, ON
THE RIGHT WE BRING TOGETHER A
LOT OF THE DATA WITH ALL THE
DIFFERENT MARKERS AND WHAT YOU
CAN APPRECIATE IS THE KIND OF
PHENOTYPE WE RESULT WITH APC-MS
SYSTEM IS PROBABLY ONE THAT
WOULD BE BENEFICIAL IN TERMS OF
OUTCOMES AND IN TERMS OF PATIENT
TREATMENTS WITH THESE CELLS AS
COMPARED TO MONOCYTE-DERIVED
DCs.
WE CAN EXPAND THE CELLS MORE
RAPIDLY, ENRICH THEM FOR
SIGNIFICANTLY AND GET CELLS THAT
LOOK LIKE THEY HAVE GREATER
FUNCTIONALITY AT THE SAME TIME.
NOW, WE ALSO HAVE LOOKED AT THE
FUNCTION AFFINITY OF THE T CELLS
BASICALLY FOR THE PEPTIDE
ANTIGEN, AND WE SEE
STATISTICALLY SIGNIFICANT
ENHANCEMENT IN TERMS OF MART1
TCR AFFINITY, ALL THE METRICS
THAT WE'VE BEEN ABLE TO LOOK AT,
AT THIS POINT IN TIME WE THINK
THIS COULD BE A NICE TECHNOLOGY
FOR EXPANSION OF TILs AND
OTHER ANTIGEN-SPECIFIC T CELLS
POPULATIONS FOR ADOPTIVE
THERAPY.
WE'VE STARTED TO DO A LITTLE BIT
OF WORK WITH BASICALLY MELANOMA
PATIENTS, IN THIS CASE TREATMENT
NAIVE MELANOMA PATIENTS.
WHERE WE BASICALLY HAVE DONE
BLOOD DRAWS, WE'VE SCREENED
BASICALLY FOR HLA2, BASICALLY
THE MHC THAT WE'RE BASICALLY
PRESENTING FROM THE APC-MS, AND
THEN BASICALLY EXPLORING WHETHER
WE CAN EXPAND MART1 CELLS FROM
THEIRS THESE PATIENTS.
ABOUT 5 OF THE 8 CELLS, 5 OF 8
PATIENTS FROM WHO WE RECEIVE
SAMPLES TO DATE WE SAW
SIGNIFICANT EXPANSION, GREATER
THAN 5% OF C -- T CELL
ENRICHMENT, A FEW GAVE DRAMATIC
RESULTS, PATIENT 11, 55% WERE
MART1 SPECIFIC.
THE GRAPH ON THE RIGHT SHOWS
DISTRIBUTION WHERE WE HAD A
PRETTY GOOD RESPONSE FROM THE
MAJORITY OF PATIENTS, WE THINK
THE STRATEGY WILL BE USEFUL
BROADLY, NOT JUST FROM THE
CONTEXT OF HEALTHY T CELLS BUT
ALSO THOSE FROM PATIENTS WITH
DISEASE, AND WE HAVE SOME
ONGOING EFFORTS NOW TO TAKE SOME
OF THE PATIENT SAMPLES FROM SOME
OF CATHY WU'S NEW ANTIGEN
VACCINE TRIALS AND SEE IF KEY
CAN EXPAND OUT SPECIFIC
POPULATIONS IN THE SAMPLES FROM
PATIENT CLINICAL TRIALS.
SO, THE LAST THING I WANT TO
MENTION ABOUT THIS SYSTEM, WE'RE
ALSO THINKING THAT, WELL, MAYBE
WE COULD USE IT BEYOND IN CELL
CULTURE TO EXPAND T CELL
POPULATIONS IN TERMS OF
MANUFACTURING, MAYBE WE COULD
ALSO USE IT TO ACTUALLY BOOST T
CELLS THAT ARE ALREADY IN THE
BODY.
AND SO THE FIRST FORAY INTO THIS
WAS WORK BY DAVID DANE, AND
JOSHUA MOVED TO HIS OWN FACULTY
POSITION, UNIVERSITY OF
WISCONSIN, JOSH BROCKMAN.
IF IT'S NOT PROVING TO BE
EFFECTIVE COULD WE JUST DO A
SIMPLE SUBCUTANEOUS INJECTION OF
PARTICLES, HAVE THE CELLS HOME
SO THE PARTICLES GET A I
APPROPRIATE STIMULATION AND
EXPAND AND ALTER PHENOTYPE OF
THESE CELLS, STRAIGHTFORWARD
IDEA.
THE FIRST EXAMPLE WE'VE JUST
COMPLETED.
THIS IS IN CONTEXT OF POLYCLONAL
T CELL EXPANSION USING THE SAME
MODEL WITH T-CELL THERAPY IN
IMMUNOCOMPROMISED ANIMALS.
UNTREATED ANIMALS SUCCUMB
RAPIDLY.
IF WE PROVIDE CAR T AND
PURPOSELY DELIVER SUBTHERAPEUTIC
DOSE TO CAR-T CELLS, SO IT WOULD
BE PARTIALLY EFFECTIVE LOSING
EFFECTIVENESS WITH TIME AND
INJECTED WITHOUT STIMULATORY
CUES THEY WERE ALSO INEFFECTIVE.
IF WE DELIVERED CUES WITH THE T
CELL, TCR STIMULATION AND
CO-STIMULATION, ALSO RELEASING
SMALL AMOUNT OF IL-2 WE GOT
GREATER RESPONSE, I'M NOT
SHOWING DATA, BUT DUE TO THE
FACT WE GOT RESTIMULATION OF T
CELLS, EXPANSION OF DELIVERED
CAR-T CELLS, AND THAT SEEMED TO
UNDERLIE EFFECTIVENESS IN THIS
KIND OF MODEL.
SO WE THINK THIS COULD
POTENTIALLY BE INTERESTING IN
THE FUTURE FROM POTENTIAL OF
DIRECT IN VIVO EXPANSION IN
POPULATIONS OF INTEREST AS
WEVMENT -- WELL.
HOPEFULLY, HOW WE CAN PROVIDE A
VARIETY OF CUES, WITH THESE
PARTICLES, WITH THIS WE CAN MORE
RAPIDLY EXPAND T CELLS, WE CAN
TUNE THE PRODUCTS, WE CAN
GENERATE HIGHER FUNCTIONALITY,
AND WE THINK THIS COULD BE
RELEVANT IN A NUMBER OF
CONTEXTS.
BEYOND T CELL BIOLOGY, OR THIS
PART OF T CELL BIOLOGY WE'VE
BEEN EXPLORING ALTERNATIVE
INPUTS, VARIETY OF DIFFERENT
TYPES OF SIGNALS WE COULD
PROVIDE FROM THESE MATERIALS,
AND THEN WE'RE ALSO BEGINNING TO
THINK ABOUT THIS AS BEING A
BROADLY USEFUL PLATFORM FOR
EXPANSION OF A VARIETY OF CELL
TYPES BEYOND T CELLS, SO, FOR
EXAMPLE, IN COLLABORATION WITH A
LABORATORY AT THE DANA-FARBER
WE'VE BEEN LOOKING AT NK CELL
EXPANSION, MANY OF YOU ARE QUITE
FAMILIAR, THEY ARE OF GREAT
INTEREST TODAY, EXPANSION OF
THESE CELLS IS PROBLEMATIC.
BUT WHAT WE'VE BEEN ABLE TO
DEMONSTRATE WITH THIS SYSTEM IN
THIS CASE PRESENTING IL-21 AND
4-1BBL, JOHN CUE -- CONJUGATED
TO THE MEMBRANE CAN GET UP TO
2,000-FOLD EXPANSION OVER THE
TIME COURSE OF A FEW WEEKS AND
CELLS WE'VE EXPANDED AGAIN STAY
HIGHLY FUNCTIONAL.
WE THINK THIS KIND OF STRATEGY,
THIS KIND OF APPROACH MIGHT BE
USEFUL IN A VARIETY OF CONTEXTS
BEYOND T CELLS.
OKAY.
SO I'D LIKE TO CHANGE GEARS NOW
AND TALK ABOUT ANOTHER ASPECT OF
THE TECHNOLOGY, AGAIN RELATED TO
T CELL BIOLOGY.
BUT HERE THE IDEA OF TRYING TO
DEGENERATE T-CELL RESPONSES
USING THERAPEUTIC VACCINATION.
THE GENERAL STRATEGY WE FOLLOW
IN THE LABORATORY SHOWN IN THIS
SLIDE, WHICH WE FIRST PROPOSED
NOW 15 YEARS AGO.
AND WHAT WE PROPOSED AT THAT
POINT IN TIME IS THAT ONE COULD
TAKE A BIOMATERIAL AND ONE COULD
PLACE IT IN THE BODY, AND IF IT
RELEASED WHAT WE CALL RECRUITING
FACTORS, IN THE CONTEXT OF MY
COMMENTS TODAY THAT WILL BE
GMC-MS, COULD DRAW IN
POPULATIONS BY CHEMOTAXIS,
AIMING TO BRING IN IMMATURE
DENDRITIC CELLS, PROVIDE
APPROPRIATE AGENTS, IN THE
CONTEXT OF CANCER AND ADJUVANTS
TO ACTIVATE CELLS, LOAD THEM
WITH ANTIGENS, AND INDUCE THEM
TO ACTUALLY THEN RETURN TO THE
DRAINING LYMPH NODES.
A KEY FEATURE HERE IS THAT THIS
IS NOT A ONE SHOT.
THIS IS A CONTINUOUS PROCESS.
WE THOUGHT OF THIS AS A CELL
FACTORY IN THE BODY, YOU'RE
CONTINUOUSLY BRINGING IN CELLS
FOR A PERIOD OF TIME, ACTIVATING
THEM, LOADING WITH ANTIGEN,
SENDING THEM TO THE DRAINING
LYMPH NODES, TO THEN INDUCE
ADAPTIVE IMMUNE RESPONSE.
THAT WAS THE GENERAL IDEA.
FIRST GENERATION DEVICE WAS
WD-VAK FROM POLYMERS USED TO
MAKE SUTURES.
A LYSATE OBTAINED FROM BIOPSY OF
TUMORS, AN ADJUVANT, A SMALL
TABLET ABOUT THE SIZE OF A BABY
ASPIRIN UNDER THE SKIN OF A
PATIENT.
WE DID ALL THE PRE-CLINICAL
STUDIES, PHASE 1 CLINICAL TRIAL
RUN BY STEVE HODE AT THE
DANA-FARBER, THIS IS STEVE WITH
ONE OF THE FIRST PATIENTS THAT
RECEIVED THE PLASTIC UNDER THE
SKIN TO TREAT MELANOMA.
I'LL LEAVE IT TO STEVE TO
PRESENT THE OUTCOME IN TERMS OF
CLINICALLY THE OUTCOME BUT FROM
AN ENGINEERING PERSPECTIVE
THERE'S TWO KEY OBSERVATIONS WE
MADE.
THAT WERE LIMITATIONS OF THIS
TECHNOLOGY.
ONE IS THAT WE HAD TO DO A SMALL
INCISION, PLACE OUT UNDER THE
SKIN.
THAT WAS OKAY BUT NOT IDEAL.
YOU'D RATHER HAVE A MINIMALLY
INVASIVE DELIVERY.
THE SECOND LIMITATION IS THAT WE
COULDN'T ACTUALLY FABRICATE THIS
VACCINE, UNTIL WE HAD THE
ANTIGEN OF INTEREST FROM THE
PATIENT.
WHICH MEANS WE DO A BIOPSY, WE
WOULD BASICALLY MORE OR LESS
FREEZE DRY IT, AND THEN MAKE THE
VACCINE.
SO THIS RESULTED IN DELAY OF
MULTIPLE WEEKS BEFORE WE COULD
TREAT A PATIENT.
SO, WE THOUGHT WE NEEDED TO MOVE
ON TO A NEXT GENERATION SYSTEM
THAT WOULD OVERCOME THESE
LIMITATIONS.
I SHOULD ALSO MENTION THIS
TECHNOLOGY WAS LICENSED BY
NOVARTIS AND IN PART BY A SMALL
START-UP.
ATAVARE.
ONE OF THE SYSTEMS IS SHOWN
HERE.
WE DECIDED TO USE THE SAME TYPE
OF MICROPARTICLES THAT I JUST
DESCRIBED THAT WE'RE USING FOR
ARTIFICIAL ANTIGEN-PRESENTING
CELLS HERE WITHOUT A LIPID
COATING, USING THESE AS MICRO
PARTICLES TO WHICH WE CAN ABSORB
DIFFERENT CARGOES ON DIFFERENT
SETS OF MICROPARTICLES, ALTERING
THE CHEMISTRY, MIX AND MATCH
DIFFERENT PARTICLES THAT MAYBE
ONE WOULD HAVE ADJUVANT, ONE
GCM-MS, ANOTHER AN ANTIGEN OF
INTEREST.
WE CAN MAKE THE SYSTEM, AND WE
HAVE THEM READY TO BE USED IN A
PATIENT AND IF WE'RE OBTAINING
ANTIGEN FROM A PATIENT WE CAN
JUST ABSORB TO THESE PARTICLES
AFTER THEY HAVE BEEN FULLY
MANUFACTURED, AND I'LL SHOW YOU
NEXT DATA FROM THAT APPROACH.
ABSORB IT FOR A COUPLE HOURS AND
INJECT, MAKE IT NEEDLE
INJECTABLE AND MINIMIZE THE WAIT
BEFORE WE CAN ACTUALLY MOVE TO
TREATMENT.
THIS IDEA WAS FIRST BROUGHT TO
FRUITION BY KIM AND LEE IN THE
LABORATORY, NOW RUNNING A
LABORATORY IN SOUTH KOREA,
EILEEN IS WORKING FOR LYELL.
LONG ALIGNED NANOPARTICLES,
NANOPORES, WE PUT THE CARGO, AND
THE IDEA HERE IN CONTEXT OF
VACCINE WE COULD HAVE THESE IN
SUSPENSION IN SALINE, WOULD
INJECT THEM, AND AFTER INJECTION
THE SALINE DISSIPATE, PARTICLES
COLLAPSE AND CREATE MICROPOROUS
MATERIAL SCAFFOLDING SIMILAR TO
WDVAX, IN THE BODY, CELLS WITH
CRAWL INTO THE PORES AS OPPOSED
TO WDVAX OUTSIDE THE BODY.
WE COULD RECRUIT LARGE NUMBERS
OF CELLS, COMPARED TO THE SAME
MATERIAL WITHOUT BLUE.
WE GET HIGH NUMBERS OF
ANTIGEN-PRESENTING CELLS AS
INDICATED BY CD11 STAINING, WE
GET A COUPLE MILLION
ANTIGEN-PRESENTING CELLS TO THAT
SITE.
AND IF WE RELEASE AN ADJUVANT,
CpG, KEY INCREASE SURFACE
EXPRESSION OF CD86, 82,
ACTIVATING DENDRITIC CELLS
SIMULTANEOUSLY LOADING WITH
ANTIGEN.
BOTTOM SHOWS THE IDEA THAT WE
CAN USE THESE AS A SUSTAINED
RELEASE SYSTEM, AS I MENTIONED
BEFORE, WE'RE WE LOADED WITH
FLUORESCENTLY LABELED ALBUMIN,
INJECTION SUBCUTANEOUS, FOLLOWED
THE SIGNAL OVER TIME.
YOU CAN APPRECIATE A COUPLE
WEEKS OF CONTINUOUS PRESENTATION
OF ANTIGEN WITH THIS SYSTEM.
NOW, WHEN WE LOOKED AT THE
RESULTING RESPONSE WE'VE
PUBLISHED THIS IN A NUMBER OF
FORMATS, SO I'M GOING TO GIVE A
COUPLE PIECES OF DATA TO GIVE A
SENSE OF THE PERFORMANCE OF THE
SYSTEM.
IN THIS CASE WHEN WE'RE USING
GnRH, A SMALL PEPTIDE, AS THE
ANTIGEN WHAT YOU CAN APPRECIATE
WITH SINGLE INJECTION, SINGLE
VACCINATION, THERE'S NO BOOST.
WITH THREE SYSTEMS, THE ORIGINAL
ONE WDVAX, A GEL SYSTEM I WON'T
DESCRIBE DATE GENERATED HIGH AND
QUITE DURABLE TITERS THAT LAST
THE FOR WELL OVER A YEAR WITH
SINGLE VACCINATION, AND ALSO
ABLE TO GENERATE PRETTY ROBUST
T-CELL RESPONSES.
HERE AS INDICATED BY T-CELL
RESPONSES WHEN USING OVA AS THE
ANTIGEN, e-7 PEPTIDE, AND
BASICALLY THE RED IS THE
OPTIMIZED VACCINE, ALSO WHEN WE
LOOK AT TILs B 16S 10 MODEL
USING NEOANTIGENS AS SOURCE TO
GENERATE THE RESPONSE, AGAIN YOU
SEE WE GET A SIGNIFICANT
INCREASE IN BASICALLY TILs
WITHIN THE TUMORS IN ANIMALS.
SO SYSTEM WORKS WELL GENERATING
HUMORAL RESPONSES, EFFECTIVE IN
MURINE MODELS, SIGNIFICANT
ADVANTAGE, FOR EXAMPLE, IN
CONVEX OF BASICALLY TC 1
SUPPRESSING MODEL OF CERVICAL
CANCER AND NICE EFFECTIVENESS
IN B 16, SYNERGISTIC EFFECT WITH
CHECKPOINT BLOCKADE THERAPY, AS
ONE WOULD EXPECT FROM REASONABLE
VACCINE.
SO THE SYSTEM BASICALLY HAS
WORKED NICELY.
AND WE'VE BEEN EXPLORING IT A
NUMBER OF CONTEXTS, ONE CONTEXT
MORE RECENTLY IS THAT MANY OF
YOU MAY BE FAMILIAR WITH THE
FACT THAT MANY TYPES OF CANCER
CELLS WHEN THEY ARE STRESSED
THEY WILL BEGIN TO EXPRESS NK
G2D LIGANDS, TARGETS FOR
CLEARANCE FROM A VARIETY OF
CYTOTOXIC LYMPHOCYTES INCLUDING
NK CELLS, MANY DEVELOPED AN
ESCAPE MECHANISM TO SHED LIGANDS
BINDING TO RECEPTORS ON
LYMPHOCYTES AND DOWNREGULATE THE
FUNCTION.
A COMMON MECHANISM OF ESCAPE
FROM MANY TYPES OF HUMAN
CANCERS, THIS HAS BEEN
DOCUMENTED.
SO KAI CAME TO THE LAB A FEW
YEARS AGO AND SAID HE WAS
COLLABORATING WITH DEVELOPMENT
OF THE SYSTEM AND PROPOSED WE
COULD USE THIS AS EFFECTIVE
MEANS OF VACCINATING
SPECIFICALLY AGAINST THE SITE AT
WHICH THESE LIGANDS ARE SHED
FROM THE SURFACE OF THE CANCER
CELLS WHICH HAD BEEN PREVIOUSLY
DESCRIBED IN THE LITERATURE.
SO WE BASICALLY USED THAT DOMAIN
OF MYC A, MYC B AS ANTIGEN IN
THE SYSTEM.
BASICALLY WE SHOWED IN THIS
PAPER THAT THIS ACTUALLY
DIMINISHED THE ABILITY OF THE
CELLS TO SHED MICA AND B.
THEY MAINTAINED IT ON THEIR
SURFACE.
SIGNIFICANTLY INCREASED
CROSS-PRESENTATION OF TUMOR
ANTIGENS BY CDCs, TARGETING OF
TUMORS BY NK CELLS AND T CELLS.
POTENTIAL POTENCY, THIS IS TWO
SETS OF DATA, BASICALLY
VACCINATING AFTER PRIMARY TUMOR
REMOVAL LOOKING TO PREVENT
METASTASES, THE TOP IN THE
CONTEXT OF B 16 MODEL, BOTTOM
WITH A 4T 1 OR BREAST CANCER
MODEL, VACCINATION HERE HAS A
SIGNIFICANT REDUCTION IN
METASTASES AFTER PRIMARY TUMOR
REMOVAL SUGGESTING THE SYSTEM IS
POTENT ABLE TO GENERATE ROBUST
RESPONSES.
WHAT WE'VE BEEN LOOKING AT THE
LAST COUPLE YEARS, WE'RE TRYING
TO UNDERSTAND HOW THIS SYSTEM
WORKS AND WHY IT'S SO EFFECTIVE.
AND ONE OF THE THINGS WE THOUGHT
AND THAT DATA SUPPORTS IS THAT
IT'S PARTLY JUST A NUMBERS GAME.
THAT THIS VACCINE BY ATTRACTING
LOTS OF ANTIGEN-PRESENTING CELLS
GETTING THEM TO MOVE TO DRAINING
LYMPH NODE CARRYING ANTIGEN,
OVER SOME SUSTAINED PERIOD OF
TIME WE GET A LOT MORE CELLS,
MOVING TO DRAINING LYMPH NODE,
MORE ACTIVITY, MORE ROBUST
RESPONSE.
WE'VE LOOKED WITH A VARIETY OF
METRICS, ALEX JUST FINISHED UP
IN THE LAB FOR EXAMPLE HAS SHOWN
THIS.
YOU CAN APPRECIATE THIS GROSSLY.
IF YOU LOOK AT THE DRAINING
LYMPH NODE, DOWNSTREAM FROM THE
VACCINATION SITE, THIS IS JUST
USING ULTRASOUND IMAGING TO LOOK
AT DRAINING LYMPH NODE WITH PBS
INJECTION, TYPICAL BOLUS VACCINE
INJECTION WITH ANTIGEN,
ADJUVANT, GMC-SF IN SOLUTION,
SIGNIFICANT EXPANSION OF THE
DOWNSTREAM DRAINING LYMPH NODE
AND EXPANSION THAT PERSISTS FOR
MULTIPLE WEEKS WITH THE VACCINE
SYSTEM.
CURRENTLY NOT SURPRISINGLY WE
SEE SIGNIFICANT INCREASES IN
DENDRITIC CELLS, T CELLS, B
CELLS, IN THE DRAINING LYMPH
NODE.
AND THE ORANGE HERE IS BASICALLY
BIOMATERIAL VACCINE, GREEN IS
BOLUS, AND THE OTHERS ARE SOME
OF THE CONTROLS.
SO, PARTLY A NUMBER GAME WE GET
A LOT MORE CELLS TRAFFICKING
ANTIGEN BACK TO DRAINING LYMPH
NODE.
BUT WE'VE ALSO BEGUN TO LOOK AT
THE GENE EXPRESSION OF THE CELLS
IN THE DRAINING LYMPH NODES.
AND STRIKINGLY WE FOUND WE JUST
DON'T HAVE MORE CELLS BUT WE'RE
DRAMATICALLY CHANGING THE
EXPRESSION, GENE EXPRESSION OF
CELLS IN THE DRAINING LYMPH
NODE.
AND SO IF WE LOOK AT JUST THE
MYELOID CELLS HERE AND LOOK AT
THE NUMBER OF DIFFERENTIALLY
EXPRESSED UP OR DOWNREGULATED
WE'RE GETTING CHANGES IN
ANTIGEN CELLS THAT ARE
IMPORTANT, INFLAMMATORY
MONOCYTES, ALSO PLASMA CELLS.
SO WE'VE BEEN DOING SOME MINING
OF DATA IN COLLABORATION WITH
SHANNON TURLEY AT GENENTECH, SO
WE FOUND INFLAMMATORY MONOCYTES
ARE DRAMATICALLY EXPANDED IN THE
MPS-BASED VACCINATION, YOU CAN
SEE THE NUMBERS ON THE TOP, ON
THE BOTTOM BASICALLY A UMAP
PLOT, WITH THE MPS VACCINATION
WE PARTICULARLY ENRICH IN CELLS
IN THIS BASICALLY ZERO QUADRANT
OF THE POPULATION, AND IF YOU
LOOK AT GENE EXPRESSION OF THOSE
CELLS YOU SEE THAT IT'S
DRAMATICALLY -- DRAMATIC
UPREGULATION IN PROCESSING, IF
YOU'RE IN GAMMA RESPONSE,
INFLAMMATORY RESPONSE, WE THINK
PART OF THE REASON IT'S SO SEE
EFFECTIVE IS THE NUMBERS BUT
PARTIALLY BECAUSE WE'RE HAVING
SIGNIFICANT ALTERATIONS IN THE
BILE GENE AND DRAINING LYMPH
NODE.
PLASMA CELLS YOU CAN APPRECIATE
WITH THIS VACCINE SYSTEM WE GET
SIGNIFICANT UPREGULATION IN THE
CELLS WITH THE BIOMATERIAL
VACCINE, FOR EXAMPLE IN IgG HG
1, AS CONTRASTED TO MORE
IMMATURE RESPONSE BASICALLY WITH
BOLUS VACCINATION.
OKAY.
SO ALL THIS WORK THAT I'VE BEEN
DESCRIBING HAS BEEN PART OF
NIH-FUNDED CENTER, NCI AND NIA
FUNDED CENTER TO ACHIEVE THESE
THINGS.
MAYBE WE'LL TALK FOR A COUPLE
MINUTES ON THE LAST PART, I'LL
TRY TO END MORE OR LESS ON TIME.
TALK A FEW MINUTES ABOUT ONE
LAST ASPECT OF THE WORK.
SO, NONE OF WHAT I'VE DESCRIBED
HOW T CELLS CAN GET INTO THE
SOLID TUMOR ENVIRONMENT WHICH WE
KNOW IS A CHALLENGE.
THIS IS JUST A REVIEW WE WROTE
LAST YEAR THAT HIGHLIGHTED A FEW
OF THE KNOWN BARRIERS TO T CELL
TRAFFICKING.
BUT WHAT WE BECAME INTERESTED IN
BEYOND ALL OF THESE THAT HAVE
BEEN PREVIOUSLY PROPOSED OR LAST
COUPLE YEARS WE'VE BECOME
INTERESTED IN CHANGES IN THE
PHYSICAL PROPERTIES OF THE
TUMORS THEMSELVES MIGHT ALSO
CREATE CHANGES IN T CELL BIOLOGY
THAT COULD BE RELEVANT IN TERMS
OF MEDIATED ABILITY TO PROVIDE
EFFECTIVE THERAPY.
NOW, THE RATIONALE HERE THAT
YOU'RE QUITE FAMILIAR WITH IS
THAT IN THE CONTEXT OF SOLID
TUMOR DEVELOPMENT, WE ALL
APPRECIATE THAT BASICALLY THE
TISSUE BECOMES STIFFER AND MORE
FIBROSED, AND THERE'S BEEN A LOT
OF INTEREST IN TRYING TO
UNDERSTAND IMPACT OF CHANGES IN
STIFFNESS THAT MAY ACCOMPANY
CANCER AND TUMOR PROGRESSION ON
BIOLOGY OF CELL TYPES.
AND PART OF THIS IS DUE TO THE
FACT THAT WHEN CELLS ENGAGE WITH
THE EXTRACELLULAR MATRIX IT'S A
MECHANICALLY ACTIVE ENGAGEMENT
WHERE THEY ARE CONSTANTLY
PUSHING AND PULLING AS SHOWN BY
THE VIDEO OF THE HUMAN IMMUNE
CELL THROUGH COLLAGEN SCAFFOLD,
ACTIVE MECHANICAL INTERACTION OF
CELLS WITH ENVIRONMENT, SO AS
MECHANICAL PROPERTIES CHANGE IT
MAKES SENSE THAT THE CELL
BEHAVIOR MIGHT CHANGE.
HOWEVER, THE EMPHASIS TO DATE
HAS BEEN ON THE IMPACT OF
CHANGES IN STIFFNESS OF THE
TISSUE.
BUT ALL THE TISSUES IN OUR BODY
AND TUMORS IN OUR BODY ARE NOT
PURELY ELASTIC LIKE RUBBER
BANDS.
THEY ARE A PROPERTY INTERMEDIATE
BETWEEN SOLID AND LIQUID.
THEY ARE VISCOELASEK.
ONE DEMONSTRATION, MAPPING OF
SKELETAL TISSUES, SOFT TISSUES,
RECONSTITUTED EXTRACELLULAR
MATRICES, THEY ALL EXHIBIT
INTERMEDIATE BEHAVIOR.
ANOTHER WAY TO DEMONSTRATE IS IF
YOU PUSH ON A VISCOELASTIC
TISSUE, IT PLASTICALLY DEFORMS
AND DISSIPATES ENERGY, STRESS
DIMINISHES WITH TIME.
YOU CAN SEE A VARIETY OF TISSUES
IN THE BODY DO THIS, IN CONTRAST
A PURELY ELASTIC MATERIAL LIKE
RUBBER BAND WOULD STAY CONSTANT
FROM THE LINE ACROSS THE TOP.
I WON'T GO DETAIL BUT A SENSE OF
WHAT IT LOOKS LIKE THIS IS A
VIDEO OF A PURELY ELASTIC
MATERIAL.
PRESS DOWN, IT DEFORMS.
WHEN YOU REMOVE, IT JUMPS BACK
TO ORIGINAL DIMENSIONS.
IN CONTRAST ON THE BOTTOM WE'RE
LOOKING AT VISCOELASTIC
MATERIAL, BRAIN TISSUE.
WHEN WE PULL UP THE TISSUE STAYS
DEFORMS, DISSIPATED SOME STRESS.
THE MOVIE CYCLES, IT JUMPS UP
BECAUSE THE MOVIE IS STARTING
OVER AGAIN.
AND SO WE BECAME INTERESTED IN
WHETHER TRANSITIONS THAT MAY
HAPPEN IN THINGS LIKE CANCER
BETWEEN A MORE VISCOELASTIC
NORMAL TISSUE AND MORE ELASTIC,
YOU KNOW, FIBROSED DEVELOPING
TUMOR MIGHT IMPACT T CELL
BIOLOGY.
A TALENTED Ph.D. STUDENT WITH
ANOTHER TALENTED Ph.D. STUDENT
IN THE LAB TO BEGIN TO EXPLORE
THIS FIRST OF ALL, WELL, LET'S
GET SOME REALITY CHECK, LET'S
LOOK AT PUBLISHED SINGLE CELL
RNAseq DATA OF T CELLS FROM
CANCER PATIENTS AND SEE IF THE
GENE EXPRESSION REALLY CHANGES
BETWEEN VISCOUS ENVIRONMENT IN
THE BLOOD, VISCOELASTIC
ENVIRONMENT IN NORMAL TISSUE,
AND MORE ELASTIC ENVIRONMENT IN
BASICALLY A SOLID TUMOR.
THEY COLLECTED DATA FOR A FEW
TYPES OF CANCER, T CELLS IN
DIFFERENT ENVIRONMENTS DID
SEGREGATE FROM EACH OTHER AND IF
YOU DEFINE BASICALLY GENE SETS,
FOR THE CELLS BASICALLY TUMORS,
STIFF ELASTIC TUMORS, YOU SEE
THEIR EXPRESSION VARIES BY
LOCATION.
THIS WASN'T CAUSE AND EFFECT,
SIMPLY CORRELATION, BUT
SUGGESTED POSSIBILITY.
I'M GOING TO SKIP THAT SLIDE
WHICH SHOWS THE MATERIAL SYSTEM
THAT ALLOWS US TO CLEANLY
REGULATE VISCOELASTICITY
SEPARATELY FROM STIFFNESS OR
VARIABLES.
WE CAN CLEANLY SEPARATE THE
VARIABLES BUT THIS MODEL SYSTEM
IN CELL CULTURE, WHICH ALLOWS US
TO CLEANLY SEPARATE
VISCOELASTICITY, PUT HUMAN T
CELLS, THEY LOOK PHENOTYPICALLY
DIFFERENT.
LOOK AT HEAT MAP ON THE BOTTOM,
T CELLS IN THE SLOWLY RELAXING
MORE VISCOELASTICITY MATERIAL
HAVE MORE ACTIVATION AND
EXHAUSTION MARKER EXPRESSION
WHILE THOSE IN THE FAST RELAXING
GELS HAVE -- EXPRESSION MORE
MARKERS RELATED TO MEMORY TYPES
OF PHENOTYPES.
FURTHERMORE IF YOU DO SOME
SINGLE CELL RNAseq ON THESE
CELLS, WHAT YOU FIND IS
EXPRESSION OF A WIDE VARIETY OF
GENES RELEVANT TO T CELL
FUNCTION SIGNIFICANTLY ALTERED
TRANSITION FROM ELASTIC TO
VISCOELASEK ENVIRONMENT.
IF YOU DO SOME ANALYSIS OF
PATHWAYS, YOU SEE AGAIN YOU HAVE
SIGNIFICANT ALTERATIONS IN
PATHWAYS BASICALLY T CELL
BIOLOGY IN THESE TWO PHYSICAL
ENVIRONMENTS AND A LOT OF THESE
ARE HIGHLY RELEVANT TO WHAT YOU
MIGHT EXPECT TO LEAD TO CHANGES
IN FUNCTION OF THE T CELLS.
NOW, TO SEE WHETHER OR NOT THIS
HAD ANY REALITY CHECK HERE, HE
TOOK THE GENE SETS HE IDENTIFIED
IN CELL CULTURE MODEL, NOW USED
THOSE AND WENT BACK TO
PATIENT-DERIVED DATA,
DIFFERENTIAL EXPRESSION IN T
CELLS IN PATIENTS, YOU
SIGNIFICANTLY MORE EXPRESSION IN
T CELLS AND PATIENT TUMORS
CONTRASTED TO OTHER
ENVIRONMENTS, DOESN'T PROVE
ANYTHING BUT SUGGESTS MAYBE
THERE'S RELEVANCE.
IN CASE YOU'RE WORRIED IT'S DUE
TO DIFFERENT T CELL POPULATION,
ACTUALLY HE THEN FURTHER LOOKED
AT OTHER SETTINGS WHERE WE KNOW
WE GET CHANGES FROM MORE
VISCOELASTIC TO ELASTIC
BACKGROUND, LIVER CIRRHOSIS,
PULMONARY FIBROSIS, SAW SAME
ENRICHMENT IN THESE GENES OF THE
T CELLS IN THESE MORE DISEASED
SETTINGS.
I'M GOING TO SKIP THIS DATA, WE
LOOKED AT THE PATHWAY, I'LL
FINISH WITH THIS LAST PIECE OF
DATA, WHICH IS WE THINK THIS
MIGHT BE RELEVANT TO UNDERSTAND
WHAT HAPPENS TO T CELLS IN SOLID
TUMORS.
ALSO THEN OPEN TO THE
POSSIBILITY MAYBE THIS IS
SOMETHING THAT WE COULD EXPLOIT
IN TERMS OF T CELL MANUFACTURING
TO CHANGE THEIR PHYSICAL
ENVIRONMENT TO GET DIFFERENT
TYPES OF FUNCTIONAL T CELLS OUT.
SO TO LOOK AT THIS, A SIMPLE
EXPERIMENT, TOOK CAR-T CELLS
THEY WERE MANUFACTURING, HAD
THEM MANUFACTURED IN A MORE
ELASTIC GEL OR A MORE
VISCOELASTIC GEL, AND
TRANSFERRED INTO THE SAME KINDS
OF MODELS, WHAT THEY
DEMONSTRATED WAS THE CELLS THAT
THEY HAD EXPANDED IN THE MORE
SLOWLY RELAXING GELS WERE MUCH
MORE EFFECTIVE IN THIS CONTEXT
IN TERMS OF CANCER CELL
CLEARANCE BACK IN THE BODY.
SO WE THINK THAT THIS MAY HAVE
RELEVANCE FOR CANCER BIOLOGY,
SOME RELEVANCE IN TERMS OF HOW
WE THINK ABOUT TRYING TO MAGNIFY
CELLS.
SO, I'M GOING TO STOP HERE AND
IF THIS IDEA OF CHANGING
PHYSICAL PROPERTIES AND
VISCOELASTICITY IS OF INTEREST
TO YOU, WE'VE SHOWN NOW IT'S NOT
JUST T CELLS THAT ARE
DRAMATICALLY IMPACTS, EPITHELIAL
TO MESENCHYMAL TRANSITION IS
INTACTED BY CHANGES IN
VISCOELASTICITY AND MONOCYTE,
ABILITY TO DO ANTIGEN
PRESENTATION ALSO DRAMATICALLY
IMPACTED BY THESE CHANGES IN
VISCOELASTICITY AS WELL.
SO I'LL STOP THERE.
I'D BE HAPPY TO TAKE ANY
QUESTIONS.
[APPLAUSE]
>> IF YOU EXPOSE THE T CELLS TO
GIVEN VISCOELASTIC PROPERTIES IS
THERE SOME MECHANICAL
(INDISCERNIBLE) DOES IT LAST?
>> THAT'S A REALLY IMPORTANT
QUESTION.
YES, THERE DOES SEEM TO BE --
YOU KNOW, T CELL MEMORY IS
DIFFERENT THAN -- THE TERM
MECHANICAL MEMORY, VERY
DIFFERENT, THERE SEEMS TO BE A
MECHANICAL MEMORY, DEPENDENT ON
TIME SCALE THE CELLS WERE
EXPOSED.
IF WE HAVE THE CELLS EXPOSED TO
THE DIFFERENT PHYSICAL
ENVIRONMENT FOR 24 HOURS, IT'S
MORE TRANSIENT AND YOU TEND TO
START LOSING THE PHENOTYPE
WITHIN ABOUT 2 OR 3 DAYS OF
REMOVING THEM.
IN CONTRAST IF THEY ARE IN THE
ENVIRONMENT FOR 3 DAYS, THEN WE
SEE THEM MAINTAINING
DIFFERENTIAL PHENOTYPE AND GENE
EXPRESSION FOR AT LEAST A WEEK,
WHICH IS AS FAR AS WE'VE LOOKED.
THEY CAN MAINTAIN IT.
IT DEPENDS ON HISTORY OF
EXPOSURE.
>> AND IF I COULD ADD A
FOLLOW-UP.
SO IF YOU TIME BASED ON NORMAL
ORGAN HOMOSTASIS DO YOU SEE
BETTER RESULT FOR YOUR CAR T
EXPANSION?
>> THAT'S A REALLY INTERESTING
IDEA.
DON'T KNOW.
WE HAVEN'T TRIED TO CAREFULLY
LOOK AT THAT YET.
YEAH, THE -- YEAH, WE DON'T KNOW
YET.
>> THANK YOU.
REALLY FASCINATING TALK.
MY QUESTION IS ABOUT YOUR --
SOMETHING YOU COVERED AT THE
BEGINNING WITH THE PROGRAMMING
KIND OF -- OR TRYING TO USE AN
A.I. ALGORITHM TO PREDICT HOW
LIKE WHEN YOU TOGGLE T CELL
INPUT IN TERMS OF TYPES OF T
CELL POPULATIONS YOU GET OUT,
WHEN YOU DO THAT HOW DO YOU
QUANTIFY LIKE WHAT CONSTITUTES A
DIFFERENT T CELL ENVIRONMENT?
AT LEAST FROM THE PROCESS YOU
SHOWED IT WAS A GRADIENT, IT'S
NOT LIKE -- HOW DO YOU KIND OF
FIGURE OUT WHICH T CELL
ENVIRONMENTS ARE SIMILAR OR NOT.
I KNOW YOU USED KIND OF
CLUSTERING, RIGHT?
HOW DO YOU KIND OF -- IS THAT --
YOU GO FROM THERE TO AN ERROR
FUNCTION OR SOME WAY OF THINKING
WHAT'S DIFFERENT OR NOT?
>> I'M NOT SURE I COMPLETELY
UNDERSTAND.
I'LL TRY.
SO WE BASICALLY COLLECTED DATA
ON A NUMBER OF DIFFERENT
PHENOTYPIC AND DIFFERENTIATION
STATUS MARKERS FOR THE T CELLS.
WE THEN BASICALLY FOR THE
STARTING CELL POPULATION, WE
THEN SUBJECTED A NUMBER OF
DIFFERENT PATIENT-DERIVED CELLS
TO DIFFERENT LEVELS OF
STIMULATION, AND KIND OF
FIGURED, OKAY, WHAT'S THE ENDING
PHENOTYPE.
AND THEN WE DEFINED -- SAID,
OKAY, IF WE WANT PHENOTYPE X, AT
THE END OF DIFFERENTIATION
PROCESS HOW DO WE NEED TO CHANGE
APC-MS TO GET US THERE AS A
FUNCTION OF THE STARTING
POPULATION?
>> THAT MAKES SENSE.
MY QUESTION WAS HOW DID YOU
FIGURE OUT LIKE WHEN YOU SAY
PHENOTYPE X, HOW DID YOU
IDENTIFY LIKE PHENOTYPE X AND Y
AS OPPOSED TO --
>> YEAH, WE BASICALLY KIND OF
TOOK WHAT WE THOUGHT REPRESENTED
THE CURRENT STATE OF KNOWLEDGE
FOR WHAT PEOPLE ARE BELIEVING
YOU WANT TO AIM FOR IN TERMS OF
ADOPTIVE CAR T THERAPY.
SO IF THIS IS WHAT PEOPLE THINK
IS THE OPTIMAL PROPERTIES, BUT
KIND OF THE APPROACH AND
ALGORITHM IS INDEPENDENT OF
THAT.
SO IF TWO YEARS FROM NOW WE AS A
FIELD DECIDE THERE IS A
DIFFERENT PHENOTYPE THAT WOULD
BE IDEAL OR DIFFERENTIATION
STATUS THAT'S IDEAL WE COULD
PLUG THAT IN INSTEAD AND
BASICALLY HAVE THIS BASICALLY
SPIT OUT TO US, OKAY, HOW DO WE
NEED -- WHAT APC-MS TO WE NEED
TO GIVE PATIENTS TO GET US
THERE.
>> THANK YOU.
>> YEP.
>> EXCELLENT TALK.
I STUDY MIGRATION OF T CELLS
FROM PERIPHERY TO CENTRAL
NERVOUS SYSTEM IN THE CONTEXT OF
AUTOIMMUNE NEUROINFLAMMATION.
THIS PROCESS IS HIGHLY
REGULATED.
ONE OF THE TOP PATHWAYS THAT'S
COME UP BEING DIFFERENTIALLY
REGULATED IN T CELLS IS EMT
PATHWAY SO INTRIGUED BY THE LAST
PART.
AP-1 PATHWAY, HAVE YOU TRIED TO
MANIPULATE THE PATHWAY IN CAR-T
CELLS BECAUSE IT'S ALSO THE
BIGGEST DRAWBACK WHERE WE TRY TO
USE CAR-T CELLS IN TREATMENT OF
BRAIN TUMORS, THEY JUST HAVE
DIFFICULTY PENETRATING THE
BLOOD-BRAIN BARRIER EVEN THOUGH
IT'S LEAKY TO INFILTRATE THE
BRAIN TISSUE.
>> YEAH, WE'VE NOT DONE THAT.
WHAT WE'VE -- ALL WE'VE DONE
FROM THAT RESPECT IS WE'VE
ACTUALLY -- WE HAVE GONE BACK TO
THE CELL CULTURE, 3D CELL
CULTURE MODEL AND HAVE
DEMONSTRATED IF WE INHIBIT THE
PATHWAY, WE INHIBIT RESPONSE TO
CHANGING VISCOELASTICITY, SO WE
DO BELIEVE WE'VE SHOWN THAT
PATHWAY IN CAUSATIVE IN TERMS OF
CELL'S ABILITY TO RESPOND BUT WE
HAVEN'T TAKEN THAT IN VIVO.
WE HAVE ACTUALLY IN THE MONOCYTE
WORK, WE HAVE TAKEN THAT BACK IN
VIVO AND DEMONSTRATED
INTERFERING WITH THE SPECIFIC
SIGNAL TO TRANSDUCTION PATHWAY
DOES ALLOW US TO SLOW DISEASE
PROGRESSION, SO IN THAT CONTEXT
WE HAVE BUT NOT IN THE T CELLS
YET.
>> OKAY.
THANK YOU VERY MUCH.
>> EXCELLENT TALK.
SO, YOU CAME FULL CIRCLE GETTING
BACK TO CAR T CELL PROLIFERATION
AND GENERATION AND
VISCOELASTICITY PLAYS A BIG
ROLE.
GOING BACK TO MSRs I ASSUME
THOSE ARE STIFF ELASTIC.
SO IS THAT A STRATEGY THAT YOU
WOULD FEED BACK INTO THAT
PLATFORM WHERE YOU'RE
CONTROLLING YOUR LIGAND DENSITY
AND THOSE SIGNALS?
>> MSRs THEMSELVES, YES, ARE
RIGID AND ELASTIC BUT WITH CELLS
ENGAGING WITH LIPID BILAYER, A
SOFT COATING CELLS SEE WHEN THEY
ENGAGE T CR WITH CUES AND
PULLING ON IT, WE HAVE FOUND AS
WE VARY THE LIPID COMPOSITION
FOR EXAMPLE USING PANEL OF
LIPIDS AT DIFFERENT MELTING
POINT, SO WE HAVE DIFFERENT KIND
OF RIGIDITY ON THE SURFACE, WE
ACTUALLY HAVE A PROFOUND IMPACT
ON THE PHENOTYPE OF THE T CELL
POPULATIONS WE GENERATE WITH
APC-MS SO WE THINK THE LAST I
GOT IS CONSISTENT WITH THAT IDEA
AS WE CHANGE BASICALLY THE
MECHANICS OF THE INTERACTION WE
CAN INDEED CHANGE THE OUTPUT,
THOUGH IT'S DIFFERENT BETWEEN
THE TWO, THE APC-MS WE'RE
DIRECTLY ENGAGING TCR,
PRESUMABLY WHAT'S SEEING
DIFFERENTIAL PHYSICAL
ENVIRONMENT WHILE IN THE WORK
WITH THE GELS THE TCR ISN'T
ACTUALLY ENGAGED WITH THE GEL,
INTEGRIN LIGANDS ARE ENGAGED
WITH THE CELL BUT BOTH SEEM TO
CONVERGE HAVING A BIG IMPACT ON
T CELL PHENOTYPE.
>> THANK YOU VERY MUCH.
>> ANY QUESTIONS OFFLINE?
NOPE?
OKAY.
WELL, A BIG ROUND OF APPLAUSE
FOR DR. MOONEY.
[APPLAUSE]
[END OF PROGRAM]