WE ARE RIGHT ON TOP OF
THE HOUR.
SO LET'S GET STARTED.
WELCOME ALL OF YOU ONLINE OR IN
PERSON.
THIS IS NCI GRAND ROUNDS AND IT
IS ABSOLUTELY MY PLEASURE TO
BRUCE TODAY'S SPEAKER DR. TIM G
RETEN, HE RECEIVED HADIS DEGREE
FROM THE UNIVERSITY OF GERMANY,
HE COMPLETED HIS RESIDENTIAL
TRAINING IN INTERNAL MEDICINE
ALONG WITH HEMEATOLOGY,
ONCOLOGY, HEPATOLOGY.
BEFORE MOVING TO NCI JOINED AS A
TENURED TRACK INVESTIGATOR AND
RIGHT NOW HE'S A CME DEPUTY
INVESTIGATOR CHIEF FOR
[INDISCERNIBLE] MALIGNANT BRANCH
AND CO-DIRECTOR FOR THE NCI
LIVER CANCER PROGRAM.
TIM IS ACTUALLY TRULY A
PHYSICIAN SCIENTIST.
HIS EARLY WORK AND DOING POST
DOC TRAINING AND JOHNS HOPKINS
RELATED TO TUMOR IMMUNOLOGY, HE
USED THIS KNOWLEDGE TOGETHER
WITH HIS MEDICAL EXPERTISE IN
THE GASTRO, ONCOLOGY, AND
HEPATOLOGY, AND MEDICAL ONCOLOGY
FOR HIS RESEARCH DEVELOP NOVEL
TREATMENT FOR PATIENT WITH LIVER
CANCER.
HE HEADS THE RESEARCH TEAM TO
STUDY THE MICROENVIRONMENT OF
CANCER IN THE [INDISCERNIBLE]
MEL ANGIO CARCINOMA.
IN,A DITION HE HELPED CONDUCT
CLINICAL TRIAL FOR PATIENTS WITH
GI CANCER, HE IS AMONG THE FIRST
1 INVESTIGATE HOW THE IMMUNE
CHECK POINT INHIBITOR WORKS IN
PATIENT WITH [INDISCERNIBLE]
WITH THE CONCEPT IN COMBINATION
TOGETHER WITH LOCAL REGIONAL
THERAPY.
WITHOUT READING HIS ACCOLADES,
MANY, MANY, MANY OF THOSE, I SAW
THE AWARD FROM NCI OUTSTANDING
MENTOR AWARD, THIS IS ACTUALLY
REALLY FANTASTIC, NIH DIRECTOR
MERITS AWARD AND NIH DEPUTY
DIRECTOR AWARD.
HE PUBLISHED MANY, MANY JOURNALS
IN THE FIELD AND HIS WORK IS
CONSIDERED TOP SIDED IN MANY,
MANY OF THOSE EMPLOY SOME OF
THOSE INCLUDE CCI, CONSIDERED AS
THE ADVANCE SCIENCE.
PERSONALLY, I THINK IT'S
EXTREMELY SMART AND VERY CLEVER
TO WORK WITH MANY PEOPLES.
SO WITHOUT FURTHER ADO, I WOULD
LIKE TO WELCOME TIM TO THE
PODIUM.
[ APPLAUSE ]
>> THANK YOU FOR THOSE VERY,
VERY KIND WORDS AND
INTRODUCTION.
SO THIS IS KIND OF THE MENU I
WILL TELL YOU, JUST TOUCH ON
HEALTHCARE DISPARITIES AND HOW
IT RELATES TO OUR PROJECTS AND
THEN I DECIDED TO BRIEFLY TELL
YOU ABOUT STUDIES THAT WE
FINISHED ONLY IN THE PAST 2 AND
HALF YEARS TOUCHING ON DIFFERENT
AREAS IN THE FIELD OF
IMMUNOTHERAPY OF LIVER CANCER,
LIVER DISEASE WE LOOK AT THE
SPECIFIC ENVIRONMENT, A LINGS
BIT OF THE MIKE ARE BIOME AND
HOW IT'S RELATED TO TREATMENT.
SO IF I START WITH THIS, THIS IS
OUR ESTIMATED DEATH RATES FOR
THE COMMON CAPSER IN THE UNITED
STATES AND [INDISCERNIBLE] FROM
THE GROUP LOOKED AT THIS AND SHE
MADE THE OBSERVATION THAT LIVER
CANCER IS THE ONLY CANCER WHERE
NONHISPANIC WHITES HAVE THE
LOWEST ANNUAL DEATH RATES IN
COMPARE TO AFRICAN AMERICANS OR
HISPANIC PATIENTS.
INDICATING THERE IS A CLEAR
DISPARITY IN THIS.
WE ASKED, YOU KNOW WHY IS THIS,
WHY WOULD THIS BE AND 1
POTENTIAL REASON SYSTEM PROBABLY
OBESITY.
SO HAD IS THE PREVALENCE OF
OBESITY IN THE UNITED STATES
AMONG NONHISPANIC WHITE ADULTS.
IF YOU COMPARE THIS TO HISPANIC
AND OBVIOUSLY THE DARKER YOU CAN
SEE THE FREQUENCY IS, YOU CAN
SEE HOW THIS SHOWS UP AND IF YOU
GO TO THE BLACK ARK DULTS, YOU
CAN SEE THERE ARE STATES WHERE
MORE THAN 50% OF THE INDIVIDUALS
DEMONSTRATE--ADULTS ARE OBESE.
NOW OBVIOUSLY WE DON'T BELIEVE
THAT OBESITY IS THE ONLY REASON
ASK WE ARE CURRENTLY STUDYING
DIFFERENT ENVIRONMENT FACTORS
AND OTHERS POTENTIALLY
IMMUNOLOGIC FACTORS THAT MAY
EXPLAIN THIS.
I THINK IT SETS THE STAGE A
LITTLE BIT YOU KNOW TO STUDY
NONALCOHOLIC FATTY LIVER DISEASE
AND THAT'S WHAT I WOULD LIKE TO
START WITH.
NOW THOSE WHO KNOW ME, KNOW THAT
WE USUALLY TRY TO ANY INTO
NICHES AND STUDY SOMETHING ELSE
NOBODY ELSE STUDIED.
WHAT I WILL START WITH IS I WILL
TELL YOU ABOUT A STUDY WE
COMPLETED, IT'S A PRECLINICAL
STUDY, A MOUSE STUDY WHERE WE
LOOKED AT PLATELETS IN THE
CONTEXT OF NONALCOHOLIC FATTY
LIVER DEC AND HCC.
NOW FROM A CLINICAL PERSPECTIVE
WE KNOW THAT PATIENTS WITH LIVER
DISEASE AND HEPATITIS B, THEY
BENEFIT FROM ASPIRIN TREATMENT.
YOU CAN SEE HERE, LARGE COHORT
STUDY OUT OF SCANDINAVIAN
CLEARLY REDUCES LIVER AND
HCC RELATED MORTALITY.
SO THIS WOULD SUGGIEST AND WE
KNOW THIS FROM ANY OTHER
DISEASE, YOU KNOW THAT
INHIBITING PLATE FUNCTION MAY BE
A GOOD THING.
NOW WE TESTED THIS IN A MOUSE
MODEL AND WE WILL SHOW YOU HOW
WE DID THIS, WE HAVE THE MOUSE
MODEL NOW HERE, WE CAN SCREEN
TUMORS SO WE INJECT TUMOR CELLS
AT 55.1 AND THEN WE PERFORM
SCANS AT DIFFERENT TIME POINTS
AND YOU CAN SEE HERE ON THIS
SLIDE, BASICALLY THE CT SCAN
FROM THE MOUSE UNFORTUNATELY WE
PUT THE MOUSE WRONG WAY AROUND
IF WE COMPARE THIS TO THE HUMAN,
YOU SEE THE FRONT AND THE BACK,
AND YOU CAN SEE THE TUMOR.
WE CAN SCAN THE MOUSE AND YOU
CAN SEE BASICALLY FROM THE HEAD
TO THE TAIL AND YOU WILL SEE
FIRST THE LUNG AND THEN YOU WILL
SEE THE LIVER OF THIS MOUSE YOU
SEE HERE AND YOU SEE HERE HOW
THE TUMOR IS ACTUALLY INSIDE THE
LIVER AND OBVIOUSLY WE CAN
MEASURE THIS IN AN EXPERIMENTAL
SETTING AND WE COMBINED THIS,
NOW WITH ELIMINATION OF
PLATEMENTS USING AN ANTIBODY GP1
B, AND INTERESTING THING HERE,
IF THIS MOUSE IS FED A DIET TO
INDUCE FATTY LIVER DISEASE, ALL
OF A SUDDEN PLATELET INHIBITION
HERE ACTUALLY CAUSES AN INCREASE
OF TUMOR GROWTH.
THIS APPLIES TO SEVERAL MODELS,
THIS IS ANOTHER MODEL WHERE WE
INDUCE THROUGH DIET, LIVER
TUMORS AND AGAIN IN THIS CASE,
NOW, WE USE A PHARMAICOLOG
CALAWAY TO INHIBIT PLATEMENT
FUNCTION AND YOU CAN SEE THAT
INHIBITING PLATELET FUNCTION
ACTUALLY PROGRESSING TUMORS AND
THIS IS NOT LIMITED TO HTC, WE
SEE THIS WITH 3 DIFFERENT LIVER
METASTASIS MODELS AND WHEN WE
GROW THESE IN THE LIVERS, THEY
ARE GROWING BIGGER.
SO NOW YOU MAY SAY, WHAT'S GOING
ON?
FIRST I SHOWED YOU PLATELETS
DECREASING INCIDENCE OF LIVER
TUMORS, AND NOW IN OUR ANIMAL
MODELS, I AM SHOWING YOU THE
EXACT OPPOSITE.
NOW I WANT TO MAKE SURE THAT I
STRESS THAT ALL OUR STUDIES ARE
DONE IN MICE THAT HAVE FATTY
LIVER DISEASE, WE DON'T SEE THE
ABSENCE OF FATTY LIVER DEC.
SO WE LOOKED INTO THIS IN MORE
DETAIL AND WE WENT AND STARTED
READING AND 1 OF THE FACTORS
THAT WE KNEW IS THAT PLATELETS
ARE ACTIVATED IN INDIVIDUALS
WITH FATTY LIVER DISEASE AND
PLATELETS ARE ALSO A MAJOR
SOURCE OF CD40 LIGAND, SO WE
LOOKED AT CD40 LIGAND IN OUR
MODEL AND INSIDE IF WE PUT MICE
ON A DIET TO INDUCE FATTY LIVER
DISEASE, CD40 IS INCREASED WITH
DIFFERENT MODEL ANDS WE ALSO
FIND THIS ACTUALLY IF WE TAKE
BLOOD FROM PATIENTS WITH FATTY
LIVER DISEASE, CD40 LIGAND IS
INCREASE THE--WE CAN DO THE SAME
THING, THIS DOES NOD APPLY TO A
MOUSE THAT DOESN'T HAVE FATTY
LIVER DISEASE AND IF WE DEPLETE
CD40, THE [INDISCERNIBLE] MAY BE
GONE.
SAME THING HERE, NOW WE USE CD40
LIGAND KNOCKOUT MICE, THERE'S NO
CENTER FOR EXCELLENCE ON AGING
ON THE VEHICLE, SO FAR, BUT IT'S
REALLY THE CD40 THAT COMES FROM
THE PLATELET THAT'S IMPORTANT
AND WE DID THIS RATHER
COMPLICATED EXPERIMENT WHERE WE
ACTUALLY INJECT MICE THAT ARE
PF4 DTR MICE, WHERE WE DEPLETE
PLATELETS, WE PUT THEM ON THE
MCD DIET SO THAT WE INDUCE SCEEZ
AND INJECT LIVER TUMORS AND THEN
WE TRANSFER CD40 LIGAND OR XOK
OUT PLATELETS AND LOOK FOR TUMOR
GROWTH.
THIS IS JUST THE CONFIRMATION WE
CAN TRANSFORM PLATELETS AND
INDEED WE SEE THE TUMORS MORE IF
WE USE A C, DID 40 LIGAND KNOCK
OUT PLATELET AS A TRANSFER
ITSELF.
WHAT IS THE DOWN STREAM EFFECT,
THE CD4 LIGAND BASICALLY
ACTIVATES KRRK DT-CELLS, WE CAN
SEE THIS HERE, IF WE PLOK THIS
T-CELL,--SORRY IF WEB CONNECTED
BLOCK THE CELLS MAKE LESS
INTERFERON-GAMMA.
IF YOU DEPLETE--IF WE DEPLETE
CD8 T-CELLS THEY MAKE LESS CD6,
AND SAME IF WE TAKE A CD40
LIGAND KNOCK OUT PLATELET.
SO THE NEXT QUESTION IS HOW DOES
THIS WORKING, WHY DOES THE PLATE
HAVE SO MUCH CDLIGAND SO WE WENT
ARK HEAD AND STARTED TO
LOOK--AND THEY HAVE CARIO SIGHTS
IN THE PRODUCTION OF CD4
LIGANDS.
FROM THIS WE USE PLASMA THAT ARE
ON AN NORMAL DIET OR MCD DIET
AND WE SHOW THAT THOSE PLATELETS
OR THOSE MEGACARRIER SITES THAT
ARE ENCUE BAITED FROM PLASMA
FROM THE MOUSE ON AN MCD DIET
ARE INDEED EPIRICHED IN KRRK D40
LIGAND AND YOU CAN SEE HERE
CORRESPONDING IMAGING.
THIS AFFECT IS DEPEND END ON
IL12.
IF YOU BLOCK IL12, YOU SEE A
REDUCTION OF CD40 LIGAND IN THE
MEGASITES.
AGAIN, WE CAN SEE THE SAME ALSO
IF WE INHIBIT THE PLATELET
FUNCTION AND AS SHOWN HERE,
SHOWN HERE, AND INTERESTINGLY,
THE PLATELETS ARE NOT ONLY
PRODUCED BY MEGACARIO CITES IN
THE BONE MARROW BUT THERE'S ALSO
A FAIR NUMBER OF MEGACARIO CITES
IN THE LUNG AND YOU CAN SEE THAT
THOSE ARE JUST EQUALLY WELL
EQUIPPED TO PRODUCE CD40 LIGAND
ENRICHED PLATELETS.
SO LET ME SUMMARIZE WHAT I'VE
SHOWN YOU HERE--THIS IS NOT
BEHAVING SO I CAN'T USE THE
POINTER FOR YOU.
SO WHAT I CAN SHOW YOU HERE IS
THAT IN THE CONTEXT OF FATTY
LIVER DISEASE, THERE IS A
RELEASE TO IL12 THAT SIGNALS TO
THE BONE AND MEGASIGHTS IN THE
BONE TO MAKE PLATELETS THAT ARE
ENRICH INDEED C, DID 40 LIGAND.
THIS PLATELETS GO BACK INTO THE
LIVER IN THE CONTEXT OF FATTY
LIVER DISEASE, SAY I ACTIVATED
THROUGH A P2 Y12 MECHANISM,
REMEMBER,OOSE INHIBITING P2 Y12,
IT ACTIVATES CD40 LIGAND ANDS
THAT BLOCKS TUMOR GROWTH.
NOW LET'S SWITCH TO THE
MICROBIOME.
I WILL SHOW YOU A STUDY WHERE WE
FINISHED RECENTLY WHERE WE LOOK
AT THE EFFECT OF THE GUT
MICROBIOME AND HOW THE GUT
MICROBIOME IN THE LIVER, SHAPES
ANTITUMOR IMMUNITY AND WITH THIS
WE WENT NOW AND LOOK AT BILLIARY
CANCER.
SO THIS IS A TUMOR THAT GROWS
FROM IN THE BIODUCT AWELL AS IN
THE IMAWL BLADDER AND THERE ARE
CERTAIN RISK FACTORS AND PRIMARY
CHOL ANGITEIS, AND BOWEL
DISEASE, BECAUSE PATIENTS WITH
THIS DISEASE HAVE A LOWERED GUT
MICROBIOME, SO IF THAT IS THE
CASE WE SHOULD LOOK INTO THIS IN
MORE DETAIL AND SEE HOW THIS
APPLIES TO A PHOSPHORYLATED
MODEL AND IF YOU CAN BETTER
UNDERSTAND THE MECHANISM FOR
THIS.
INDEED IF WE INDUCE PSC IN MICE,
WE SEE A LEAKY GUT, WE SEE THE
FOUNDATION OF OCCRUDIN IN A
GRAPHS, USING A BIODUCT
LOCATION, IF THIS HAPPENS WE
ALSO SEE AN INCREASE OF IL17 IN
THE SMALL INTESTINE, WE SEE AN
INCREASE OF SRNA AS A BACTERIAL
READ OUT IN TRANSLOCATION IN THE
PORTAL VEIN, AND WE IF WE LOOK
AT PEASHT SAMPLES WITH
CIRRHOSIS, WE CAN FIND
POTENTIALLY BACTERIA IN THE
GROWTH.
WHAT HAPPENS IN THE LIVER.
WE LOOKED AT DIFFERENT IMMUNE
CELLS AND WE FOUND IF WE INDUCE
PSC, IN THIS CASE, WE USE MDRD2
KNOCK OUT, THERE'S AN INCREASE
IN SO CALLED MYELOID DEPRESSOR
CELLS WHICH WE CAN FURTHER
CHARACTERIZE AND DIVIDE INTO
MONOSITTIC AND [INDISCERNIBLE]
WHICH ARE INCREASED IN THE LIVER
JUST BY INDUCING THE CIRRHOSIS
IN THE LIVER.
IT ALSO APPLIES TO THE SECOND
MODEL, THIS MDSCs EXPRESS
CXCR2.
A CHEMOKINE RECEPTOR THAT IS
SPECIFIC FOR CXCR1.
AND INDEED WE ACTUALLY FIND
INCREASED CXCR 1 BOTH WHEN WE
INDUCE SCLEROSIS AND CHOL
ANGITEIS, AND BOTH LEADING TO AN
INCREASE OF 16 SRNA AND BEING A
RISK FACTOR FOR CHOL ANGIAL
CARCINOMA EMPLOY IF YOU BLOOK
THIS, IN THIS CASE WE USE AN
ANTIBODY AGAINST CXCR1, IN ALL
OF THESE CASES, WE REDUCE THE
NUMBER OF THIS IMMUNOSUPPRESSIVE
MYELOID CELLS.
AND BY DOING SO, AND THIS
MECHANISM IS DEPENDENT ON TLR4
EXPRESSION ON HEPATOCYTES AS YOU
CAN SEE ON THE LOWER RIGHT.
SO THIS IS THOT REALLY A CANCER
MODEL YET, SO WHAT HAVE I SHOWN
YOU SO FAR IN WHAT I'VE SHOWN
YOU IS IN THE CONTEXT OF LIVER
CIRRHOSIS, THERE A LEAKY GUT,
WHICH LEADS TO HEPATOCYTES
MAKING CXCR1, AND THEN THE IPT
GREATER CREASE OF THE CXCR1
INCREASE IN THE DISEASE, AND
THIS MECHANISM IS TLR4, AND WE
CAN SHOW THIS IS DEPENDENT ON
THE NEGATIVE BACTERIA AND AGAIN
THIS ONLY EXPLAINS CIRRHOSIS AND
MDCs BUT NOT A TUMOR MODEL SO
WE NEXT WENT ARK HEAD AND
COMBINED THIS WITH THE TUMOR
MODEL.
SO FOR THIS AGAIN, WE USE THE
[INDISCERNIBLE] AND WE PERFORM
HYDRODYNAMIC INYEKS AND YOU CAN
SEE IF WE DO THIS, THESE MICE
ARE MORE LIKELY TO DEVELOP
TUMORS.
CAN YOU SEE THE MICROSCOPIC
PICTURES AND MICE WITH PSC, IN A
SECOND MODEL WHERE WE USE THE
MDR2 KNOCKOUT MICE, YOU SEE THE
SAME THING AGAIN.
NOW IS THIS AGAIN DEPENDENT ON
CHANGES IN THE GUT MICROBIOME
AND IS THIS DEPENDENT ON
POLYMORPHIN AMERICA DSCs, IF
WE DEPLETE THESE MDSCs, USING
AN ANTIBODY AGAINST LYSX1, OR IF
WE OVER EXPRESS CXCL1, CAN YOU
SEE IT AND INDEED THIS CONTROLS
TUMOR GROWTH.
NOW HOW CAN WE SHOW THIS IS
RELATED TO THE GUT MICROBIOME?
THE WAY WE DID THIS WAS BY USING
OR CONDUCTING FMT STUDIES.
SO IN THIS CASE, WE CONDUCT
STOOL SAMPLES FROM MICE THAT
HAVE BEEN TREATED WITH
ANTIBIOTICS WITH VANCOMYCIN, OR
[INDISCERNIBLE], AND THE
MECHANISM WE REPORT IS TLR
DEPENDENT AND INDEED IF WE TREAT
THESE MICE WITH NEOMICE IN, WE
SEE FEWER TUMORS SUGGIESTING
THIS MECHANISM OF TUMOR FROAGHT
DEPENDENT ON THIS TLR4 AND WE
CAN SHOW THIS ALSO USING
CONDITIONAL KNOCKOUT MICE WHERE
WE ACTUALLY ON KNOCK OUT THE
TLR4 MOLECULE ON HEPATOCYTE.
YOU CAN SEE THIS ON THE LOWER
GRAPH WITH THE BLUE BAR.
THERE IS FEWER TUMORS AND THEN
WE CAN RESCUE THIS IF WE
OVEREXPRESS CXCL1.
THE DOWN STREAM CELL THAT IS
AFFECTED IS MAINLY AN ENCASE O.
NOW WHAT IS THE MECHANISM THAT
WE PROPOSE HERE, SO, THERE IS
THIS INTERACTION THAT WE CALL
THE GUT LIVER ACCESS AND UNDER
NORMAL CIRCUMSTANCES YOU HAVE
THE LIVER RELEASING AND TALKING
TO THE GUT AS WELL THE AS GUT
TALKING TO THE LIVER.
ONCE YOU DEVELOP FIBROSIS OR
CIRRHOSIS, WHAT HAPPENS IS THAT
YOU WILL ACTUALLY HAVE CHANGES
IN BILE, AND YOU WILL HAVE
DISBIOSEIS, SO THE GUT
MICROBIOME IS CHANGING.
WE STUDIED THIS.
I DIDN'T SHOW YOU THIS FOR TIME
REASON.
BUT WHAT I SHOWED WAS THE LEAKY
GUT, REMEMBER THE HISTOLOGICAL
PICTURES AND THE INCREASE OF LPS
AND 16 SRNA, BACTERIA CAN GO AND
IMET INTO THE GUT EPITHELIUM AND
THEN GO THROUGH THE PORTER VEIN,
ONCE THEY REACH THE LIVER, THEY
INDUCE CXCL1 CREATION BY
HEPATOCYTES, THIS IS THROUGH A
TLR4 DEPENDENT MECHANISM,
LEADING TO ARK KIEWMULATION OF
IMMUNOSUPPRESSIVE CXCLPOSITIVE
MDSCs WHICH IN TURN BLOCK NK
CELL FUNCTION AND THEREBY
PROMOTE TUMOR GROWTH.
SO THIS IS 1 STUDY WHERE YOU
BASICALLY--SEE AGAIN HOW THE
ENVIRONMENT, THE LIVER SPECIFIC
ENVIRONMENT IS AFFECTED BY THE
GUT MICROBIOME BUT WHAT IS
ACTUALLY THE LIVER SPECIFIC
ENVIRONMENT AND I THINK THIS IS
PROBABLY THE MOST INTERESTING
PART, IF YOU WORK IN LIVER
CANCER IMMUNOLOGY WHICH IS THAT
THE LIVER IMMUNE
MICROENVIRONMENT IS VERY, VERY
DISTINCT.
WHICH BRINGS ME TO THE NEXT
TOPIC, THE LIVER IMMUNE
MICROENVIRONMENT AND FOR THOSE
OF YOU WHO DON'T LIKE T-CELLS
AND DO NOT WORK WITH IMMUNE
CELLS ALL THE TIME, LET ME
INTRODUCE YOU WHAT WE ARE
TALKING ABOUT.
SO YOU ALL KNOW CD8 T-CELLS
PROBABLY THAT RECOGNIZE MHC
CALCIUM 1 MOLECULE AND AN
ANTIIEN DERIVE FRIDAY
INTRACELLULAR PROTEINS AND
RECOGNIZE THEM ON AN MHC
MOLECULE EMPLOY NOW ARK PART
FROM THESE CLASSICAL T-CELLS WE
FIND IN THE LIVER, A NUMBER OF
OTHER IMMUNE CELLS.
WE FIND GAMMADELTA T-CELLS, WE
FIND NKT-CELLS.
THIS IS KILLING ME HERE.
SO WE FIND NKT CELLS AND WE FIND
MAIT CELLS, THIS RECOGNIZES THE
MOLECULE, RECOGNIZES THE FLIEK O
LIPID, MAIT CELLS WHICH
RECOGNIZE RIBOFLAVIN WHICH IS A
VITAMIN B METABOLITE AND A
GAMMADELTA T-CELL WHICH
RECOGNIZE FOSTER NURSED FOCUSED
ON ANTIGENS.
APART FROM THESE THERE ARE
RELATIVELY 1 CELLS AND OBVIOUSLY
I ALREADY ALLUDED A LITTLE BIT
TO THESE NK CELLS.
NOW THIS MAY BE STILL STRAIGHT
FORWARD, UNFORTUNATELY I HAVE TO
TELL YOU IT IS NOT THAT STRAIGHT
FORWARD AS YOU THINK BECAUSE WE
CAN SUBCATEGORIZE THESE CELLS ON
TRANSCRIPTION FACTORS A LITTLE
BIT MORE.
BUT WHAT I WOULD REALLY LIKE TO
TALK ABOUT IN THE REST OF MY
TIME HERE IS BASICALLY I WOULD
LIKE TO TALK ABOUT REGULATORY
T-CELLS TOWARDS THE END OF MY
TALK AND BEFORE I GET THERE,
ABOUT MAITs, WHICH ARE VERY
ABUNDANT IN THE LIVER AND CAN AS
I WILL SHOW YOU EXERT VERY
POTENT IMMUNITY AS I HAVE SHOWN
YOU IN PRECLINICAL MOUSE MODELS.
SO WE DECIDED TO STUDY THESE
MAITs WHICH ARE VERY COMMON
AND ABUNDANT IN MOUSE LIVERS IN
MORE DETAIL.
THE WAY WE DID THIS IS BASICALLY
SHOWN HERE, SO, WE SET UP A
COLLABORATION WITH OUR
COLLEAGUES AT JOARMING TOWN, THE
TRANSPLANT CENTER, AND THEY
EXPLANTED LIVERS AND WE WERE
ABLE TO CLICK HEART ATTACK
CC SAMPLES FROM THESE PATIENTS.
WE WERE ABLE TO COLLECT A PIECE
THAT BASIC LYE CONTAINS THE
ADJACENT LIVER, THE TUMOR
BORDER, AND THE TUMOR CORE AND
WE USED THIS TO ISOLATE
MONONUCLEAR CELLS AND SORT THEM
BUT MORE IMPORTANTLY WE USE THEM
FOR TISSUE EMBEDDING AND
STAINING.
THEN WE PERFORMED CODE
EXPECTATIONS SAMPLING USING
SINGLE CELL ANTIBODIES AND
SINGLE CELL FLOW CYTOMETRY AND
THE SINGLE CELL DATA WAS FURTHER
STUDIED USING BIOINFORMATIC
ANALYSIS.
NOW THIS DATA THIS DATA IS
INTERESTING, BUT WE ONLY USE IT
TO GEPERATE A HYPOTHESIS AND
DECIDED WE WILL LIKE TO
VALENTINED DIADIC FINDINGS
TRUSTEES THIS USING DIFFERENT
METHODS SO WE USED PATIENTS WE
HAD TREATED WITH IMMUNE CHECK
INHIBITORS AND SINGLE CELL DATA.
WE VERIFIED SOME OF THE DATA
USING DATA FROM PATIENT COHORTS
THAT WERE AVAILABLE AND LOOKING
AT SURVIVAL ANALYSIS, BUT MOST
IMPORTANTLY WE ALSO PERFORMED
FUNCTIONAL STUDIES IN MICE TO
REALLY VERIFY WHAT WE SEE USING
THIS HIGHLY COMPLEX DATA REALLY
TRANSLATES INTO THE MECHANISM SO
NOW NEXT DATA, I HAVEN'T DONE
ANYTHING, BUT THIS IS A COMPLEX
PROJECT THAT REQUIRED A LOT OF
COLLABORATION AND WE ARE THE KEY
PLAYERS FOR THIS WORK IT
STARTING WITH WILL ALEX CORMA AT
GEORGE TOWN, WE HAD
[INDISCERNIBLE] WHO HELPED US
WITH CODEX AND [INDISCERNIBLE],
AND IN MY GROUP [INDISCERNIBLE]
WHO LED THIS ROG ECTOMYOSIN.
SO IF YOU LOOK AT MAITs, THE
FIRST THING WE NOTICE IS IF WE
COMPARE THE NONTUMOR VERSUS
TUMOR TISSUE, THE MAITs DON'T
MAKE IT INTO THE TUMOR.
THERE'S A DECREASE IN
INFILTRATION.
WE SEE EXACTLY THE SAME IN MICE.
YOU SEE HERE THE DATA THE
NONDATA VERSUS TUMOR, FEWER M,
ITs, IN THE TUMOR, IT'S NOT
THAT THEY'RE LESS FREQUENT,
THEY'RE ALSO LESS FUNCTIONAL AS
AN EXAMPLE OF INTERFERON-GAMMA.
SO THIS IS OBVIOUSLY SOMETHING
THAT YOU KNOW THAT WE WOULD NOT
NECESSARILY LIKE.
SO LET ME JUST SUMMARIZE BEEF
BRAE, MAIT CELLS ARE ABUB ANT IN
THE LIVER DATA, I HAVE NOT SHOWN
YOU ALL THE DATA, THIS IS SOME
OF THIS, WE WERE ABLE TO MAKE
SURE IT CORRELATES WITH THE
PATIENT'S OUTCOME, I'VE SHOWN
YOU THEY CAN IMPAIR--ACTUALLY I
HAVEN'T SHOWN YOU THAT THEY
IMPAIR TUMOR FROAGHT, BUT YOU
HAVE TO BELIEVE IT, WE HAVE DONE
THE STUDIES AND THEY ARE
CHARACTERIZED BY FILTRATION INTO
LIVER TUMORS INCREASING
DYSFUNCTION AND LOSS OF TOXICITY
WITH WITHIN THE HCC TUMOR
ENVIRONMENT.
SO THIS BRINGS UP THE QUESTION,
WHY IS THIS?
WHAT CAUSES THIS INNATE CELL
DYSFUNCTION?
WHY DON'T THEY GO INTO THE
TUMORS AND WHAT MAKES THEM
DYSFUNCTIONAL IN SO IN ORDER TO
DO THIS WE DECIDED TO DO THE
CODEX ANALYSIS TO IDENTIFY IPT
GREATERUNE CELL SUBSETS
THROUGHOUT THE LIVER.
THE WAY WE DO THIS IS BASICALLY,
YOU KNOW WE DIVIDE THE CELLS
INTO CD45 POSITIVE IMMUNE CELLS
AND PAREN CHIMAL CELLS.
WE LOOK AT DIFFERENT NOT P A
RACHYMALL CELLS, THE MARKERS,
HEPATOCYTES, ENDOTHELIAL CELLS
AND VASC VASCULAR ENDOTHELIAL
CELLS, AND WE LOOK AT THE CELLS
ON TOP OF THE LIVER EMPLOY WE
SELECTED A GROUP OF MARKERS THAT
CAN HELP US ASSESS THE FUNCTION
OF THE DIFFERENT IMMUNE CELLS
EMPLOY SO WITH ALL THIS DATA,
YOU GET YOU GET A LOT OF
INFORMATION BUT BEFORE WE GO
THERE I WILL LIKE TO SHARE THE
PRIMARY IMAGES ACTUALLY THAT WE
WERE ABLE TO OBLIGATIONS STAIN.
SO THIS IS AN EXAMPLE OF THIS,
YOU CAN SEE ON THE SLIDE.
ON THE BOTTOM RIGHT IT IS
BASICALLY THE TUMOR CORE, IN
YELLOW, CAN YOU SEE RIM OF THIS
TUMOR AND IN RED THE AREA OF THE
NONTUMOR TISSUE AND SINCE THIS
IS A MICROSCOPIC PICTURE, YOU
CAN ZOOM IN AND SEE THE
PERIPORTAL AREA AND RECOGNIZE
THE DIFFERENT STRUCTURES THAT
YOU FIND IN THE NONTUMOR LIVER.
NOW, OBVIOUSLY, WE HAVE WAY TOO
MANY MARKERS AS YOU CAN SEE ON
THE SIDE THAT I CAN SHOW YOU ON
1 PICTURE, SO LET ME SHOW YOU
HOW THIS LOOKS ACTUALLY IN
REALITY, SO THESE ARE THE
DIFFERENT MARKERS THAT WE USED
HERE.
YOU CAN SEE IF WE TART JUST
BASICALLY TURN THE LIGHT ON TO
LOOK FOR KRRK DECKER 45 POSITIVE
CELLS, OR IF WE LOOK FOR LIVER
SIGNUE SOID CELLS, IF WE LOOK
FOR MACROPHAGES THROUGHOUT THE
LIVER, IF YOU ARE INTERESTED IN
THE T-CELLS, YOU CAN FIND THEM,
THE GRANUE LO SIGHTS OR FOR
INITANCE IF YOU WANT TO TODAY
B-CELLS YOU CAN SEE WHERE
THEY'RE LOCATED.
NOW ONCE YOU HAVE THIS DATA
TRUSTEES THESE PATIENTS,
OBVIOUSLY THERE IS A LOT OF WORK
DOWN STREAM, SO SO AFTER
STAINING THIS AND GET
THANKSGIVING ON THE MICROSCOPE,
YOU HAVE TO PERFORM TISSUE
ANNOTATION SO YOU HAVE TO
DETERMINE WHICH AREA OF THE
SLIDE IS TUMORRED AND HERE I
WOULD LIKE TO ACKNOWLEDGE THE
WORK FROM DAVID CLIENER WHO
HELPED US WITH THIS.
THIS IS JUST ANOTHER EXAMPLE TO
SHOW YOU FROM ANOTHER TISSUE TO
SEE YOU HAD THIS LOOKS LIKE
EMPLOY NOW HERE YOU SEE, SIMILAR
TO SINGLE CELL DATA WE CAN THEN
PERFORM IMMUNE ANALYSIS AND AND
CHARACTERIZE THE'MUNE CELL
SUBSETS ON THE HEAT MAP EMPLOY
THE INTERESTING PART AGAIN, IS
THE PART AFTERWARDS WHERE YOU
ACTUALLY PERFORMED THE
NEIGHBORHOOD ANALYSIS AND IN
THIS CASE I HAVE TO GIVE A BIG
COLLABORATORS THAT WE HAVE THAT
DEVELOPED--NOT NOVEL BUT
PROTOCOL OR CODE TO PERFORM THIS
NEURAL NETWORK ANALYSIS WHICH IN
THE END TOLD US THAT IF WE TRY
TO FIND OUT WHICH IS THE MOST
LIKELY CELL THAT IS CLOSE TO A
MAIT, IT TURNED OUT TO BE A PDL1
POSITIVE TUMOR ASSOCIATED
MACROPHAGE.
NOW THIS IS BASED ON COMPUTER
ALGORITHM IN THE DATA WE FED
INTO THE SYSTEM BUT BECAUSE WE
HAVE IMAGES WE CAN ACTUALLY GO
BACK AND SEE, IS THIS TRUE IN
AND INDEED WE FIND THE MAITs
NEXT TO THE CDL1 OR 6 POSITIVE
MACROPHAGE.
SO THIS IS AN IMMUNE CELLAT
LAS PINTAS, I FOCUSED ON THE
MAITs AND THEY ARE CLOSE
PROXIMITY TO THE MA ROUGH ATOM
PHAGES IN THE TUMOR SURROUNDING
TISSUE.
BUT THIS IS STILL DESCRIPTIVE.
WE HAVE A LOT OF DATA, I DIDN'T
SHOW YOU ALL THE SEQUENCING DATA
WE HAVE BUT WE STILL DON'T KNOW
BUT WE DON'T KNOW IF THIS IS
REALLY FUNCTIONAL.
SO WE DECIDED THAT FUNCTIONAL
STUDIES ARE REQUIRED TO REALLY
UNDERSTAND THE BIOLOGICAL
SIGNIFICANCE OF THIS INTERACTION
OF MAITs AND PDL1 POSITIVE
TEMs, SO HOW DID WE DO THIS?
WE ACTUALLY DID AND I WILL NOT
SHOW YOU ALL OF THIS BUT WE TOOK
6 DIFFERENT APPROACHES, SO WE
PERFORMED STUDIES USING HUMAN
DERIVED SAMPLES WHERE WE CAN
ACTUALLY CO INCUBATE THEM XI
WILL SHOW YOU THIS.
I HAD ALREADY SHOWN YOU THAT WE
HAVE MOUSE MODELS WE CAN USE
THAT WE CAN LOOK AT USED TO
STUDY MAITs, SO WE PERFORMED
IN VIVO STUDIES WHERE WE BLOCK
PDL1 INTERACTION.
WE WENT BACK TO SINGLE CELL
ANALYSIS FROM HUMAN AND TUMOR
BEARING MICE.
WE PERFORM DEPLETION TODAYS
BECAUSE WE HAVE 2 CELLS THAT
INTERACT WITH EACH OTHER, IT'S
EASY TO TAKE 1 OUT AND SAY WHAT
HAPPENS CAN WE SEE AN EFFECT?
WE CAN BE EVEN MORE SPECIFIC AND
USE CONDITIONAL KNOCKOUT MICE
WHERE THE PDL1 IS SPECIFICALLY
ELIMINATED ON MACROPHAGE SUBSETS
AND WE HAD LIMITED DATA
AVAILABLE FROM OUR OWN PATIENTS.
TREATED WITH CHECK POINT
INHIBITORS WITH ANTIPDL1 TO SEE
WHETHER WE SEE AN EFFECT SIMILAR
TO WHAT WE WOULD PREDICT
PRACTICES OUR MOUSE STUDIES AS
WELL AS HUMAN STUDIES.
SO LET'S START WITH THE FIRST 1,
WE TOOK TISSUE FROM SPECIMEN,
PRIMARY PATIENTS WITH HCC, AND
WE ISOLATED MONONUCLEAR CELLS
AND ISOLATED THEM TO
TETRAMERPOSITIVE MAITs AS WELL
AS TUMOR ASSOCIATED MA ROUGH
ATOM PHAGES AND WE COLOR THEM
AND THE RESULTS ARE SHOWN HERE.
IF WE COCULTURE THE SALES, THEY
ARE INNATE THIS THEIR 80 TO
PRODUCE INTERFERON-GAMMA, AS YOU
SEE ON THE LEFT, THE CONTROL IS
GOING DOWN AND THEY HAVE NO
EFFECT EMPLOY THIS IS INDEED
CONTACT DEPENDENT.
IF WE PUT THE SAME EXPERIMENT IN
A TRANSFER WHERE WE PUT A
MEMBRANE BETWEEN THE CELLS, WE
DON'T SEE THE EFFECT ANYMORE, WE
SEE INTERFERON-GAMMA IS GOING
UP, AND THE SAME APPLIES TO AN
EXPERIMENTAL SETTING WHERE WE
NOW USE AN ANTIBODY AGAINST PDL1
SO THAT THIS INTERACTION IS
BLOCKED AND AGAIN THE FUNCTION
OF THESE MAITs IS INKRESSED
AND INTERFERON-GAMMA IS GOING
UP.
O THIS IS FIRST INDICATION THAT
INDEED THIS INTERACTION MAY BE
AT LEAST RELEVANT FOR THE
DYSFUNCTION OF MARK ITs.
HOW ABOUT AN IN VIVO EXPERIMENT?
SO NEXT WE WENT AHEAD AND USED
TUMOR MODEL THAT WE GENERATED
WHERE WE INJECT TUMORS STRAIGHT
INTO THE LIVER AND WE TREAT THEM
WITH PDL1.
AND INDEED IF WE DO THIS YOU CAN
SEE ALL OF A SUDDEN MAITs
INCREASE UPON PDL1 TREATMENT,
IT'S NOT ONLY THAT MAITs GO UP
BUT THE OTHERS GO UP AS WELL.
SO FOR THOSE WHO ASKING ME WHY I
USE THE MC38, I AM SHOWING YOU
WHERE WE USE THESE TUMORS.
THEN WE WENT AHEAD AND LOOKED AT
MORE DETAIL AT THE HUMAN DATA
THAT WE HAD AND MORE IMPORTANTLY
AT THE MYELOID CELLS.
I TOLD YOU THAT WE PERFORM
SINGLE CELL ANALYSIS WHERE WE
IDENTIFY THE DIFFERENT B-CELLS,
T-CELLS AND THE MYELOID CLUSTER.
WE ZOOMED INTO THE MYELOID CLUOF
THENER MORE DETAIL AND FOUND
THERE WERE ACTUALLY 6 DIFFERENT
CLUSTERS THAT WE COULD IESHES
DENTIFY.
ONE OF THESE CLUSTERS, WAS
ENRICHED IN CD163, THAT'S THE
SECOND CLUSTER.
THESE ARE THE TUMOR ASSOCIATED
MACROFAINLS WHICH ALSO EXPRESS
206.
WE LOOK AT THESE CELLS IN MORE
DETAIL AND WHAT WE WERE ABLE TO
SEE IS THAT IF WE LOOK FOR THE
CD163 CDLB MACROPHAGES WE FIND
THEM IN THE TUMOR TO BE
INCREASED IF WE COLLECT SASM
ETCHS FROM PATIENTS WHO HAVE
HCC, VERSUS THOSE PATIENTS THAT
HAVE OTHER LIVER DISEASES BUT
NOT A TUMOR.
AND THIS INDICATES THAT THE
PRESENCE OF A TUMOR ACTUALLY
SIGNALS TO THE ENVIRONMENT AND
LEADS TO THIS ACCUMULATION OF
TUMOR ASSOCIATED MACROPHAGES
WHICH INEXPERT IMMUNOSUPPRESSIVE
FUNCTION AND 1 OF THE MECHANISMS
IS BY UPREGGULING THE PDL1 AND
IT'S UPREGULATED ON THESE
MACROPHAGES.
NOW BASED ON THIS DATA, WHERE WE
SHOWED THAT IT'S INDEED THE PDL1
EXPRESSION ON THESE MACROPHAGES
AND WHAT I AGAIN DOESN'T SHOW
YOU THERE ARE CSFR POSITIVE
MACROPHAGES, WE WENT TO THE NEXT
STEP, AND CONDUCTED STUDIES IN
MICE WHERE WE NOW SPECIFICALLY
KNOCK OUT THE PDECKER L1 ON THE
MOUSE MACROPHAGES AND YOU CAN
SEE THIS WELLS TO AN INCREASE OF
MAITs, AND AN INCREASE OF
INTERFERON-GAMMA AS WELL AS
INCREASE IN GRANZYME PRODUCTION,
THIS IS ALSO SHOWN ON THE KNOCK
OUT ON THE DENDRITIC CELLS BUT
HERE WE DON'T SEE THE SAME
EFFECTS ON THE CSFR+ TAMs.
LAST BUT NOT LEAST, WE WERE
ASKING CAN WE USE THESE MAITs
TO ACTIVATE IMMUNE RESPONSE IN
COMBINATION OF PDL1 THERAPY.
IMEAN WE USED AN ANIMAL MODEL,
AND WE INYECT THE TUMOR CELL AT
55.1 WHICH IS AN HKRRK C CELL,
IN A WILD-TYPE MOUSE OR DOESN'T
HAVE ANY MAITs, AN MR1 KNOCK
OUT MOUSE, WE USE 5 PRU AND CPG
WHICH ACTIVATES MAITS AND WHAT
YOU CAN SEE IN THE MIDDLE GRAPH
AND THE WILD-TYPE MICE AND WHILE
WE DON'T FIND AN EFFECT IN THIS
PARTICULAR MODEL USING ANTIPDL1
EITHER AS A SINGLE AGENT OR TO
ACTIVATE T-CELLS OR THE 5 OF PRU
AND CPG, TO ACTIVATE THE
MAITs, AND THE COMBINATION
INDEED IS LEADING TO A REDUCTION
OF TUMOR WEIGHT OR TUMOR GROWTH.
CAN THIS MECHANISM IS INDEED
DEPENDENT ON MAITs, BECAUSE IF
YOU USE AN MR1 KNOCK OUT MOUSE,
THERE IS BASICALLY NO EEIVET
LEFT.
SO I BELIEVE WHAT I WAS ABLE TO
HOPE--YOU KNOW WHAT I WAS ABLE
TO SHOW YOU HERE IS THAT THESE
CELLS WHICH ARE VERY ABUNDANT
ABOUT AT THE SAME FREQUENCY THAN
CD8 T-CELLS IN THE LIVER, THEY
CAN ACTUALLY EXERT VERY POTENT
ANTITUMOR IMMUNITY.
THEY ARE COMPROMISED IN THE
CONTEXT OF A TUMOR AND THE
MECHANISM THAT WE PROPOSE IS
BEASKLY SHOWN HERE.
SO WE BELIEVE THAT WHEN THE
CELLS ARE IN THE ADJACENT LIVER,
THEY INTERACT WITH CD163
POSITIVE TUMOR ASSOCIATED
MACROPHAGES THROUGH PD1, PDL1.
THIS LEADS TO A DECREASE IN THE
INFILTRATION IN THE TUMOR CORE
AND A DECREASE OF THEIR FUNCTION
EMPLOY THEY BECOME
DYSFUNCTIONAL.
I'VE SHOWN YOU SOME OF THE READ
OUTS, I'VE SHOWN YOU A DECREASE
IN INTERFERON-GAMMA, A DECREASE
OF GRANZYME B, I DID NOT SHOW
TNF ALPHA BUT WE DID LOOK AT IT,
I DIDN'T SHOW IT.
I'VE SHOWN YOU CONDITIONAL KNOCK
OUT IS ITS, I'VE SHOWN YOU
STUDIES WHEREY WOO BLOCK PDL1,
USING ANTIABOUTS, I DIDN'T SHOW
THE DATA WHERE WE BLOCK CSFR,
BUT IT'S A SIMILAR MECHANISM
WHERE WE DEPLETE THESE
MACROFEAJS USING OTHER MODELS WE
HAVE AVAILABLE, THIS LEADS TO AN
INCREASE OF THE FUNCTION,
INCREASE IN INTERFERON-GAMMA,
GRANZYME, AND THE TUMOR, AND THE
TUMOR BECOMES ENFIL TRAITED BY
MAITs AND BECOMES MORE
INFLAMED AND ULTIMATELY SHRINKS
IN TUMOR SIZE EMPLOY NOW WHAT I
HAVE AS A TITLE IN MY TALK WAS
FROM BENCH TO BEDSIDE AND BACK
AGAIN.
AND I THINK WHAT I'M TRYING TO
SHOW YOU IS, AND THAT'S REALLY
THE THEME OF WHAT I AM TRYING TO
DO AS THE THEME OF THIS TALK IS
TO SHOW YOU HOW CAN WE COMBOYNE
HUMAN DATA AND MOUSE DATA TO GET
REALLY THE MAXIMUM INFORMATION
OUT OF IT.
AND WHAT I WOULD LIKE IN THE
LAST 10 MINUTES OR LESS TO
ACTUALLY SHARE WITH YOU SOME
UNPUBLISHED DATA.
THIS IS ALTHOUGH IT'S RECENTLY
PUBLISHED DAILY BASIS THEA ISSUE
UNPUBLISHED DATA WHERE
NONAPOPTOTIC I FEEL REALLY PUSH
THIS TO THE EXTREME AND IT'S
OBVIOUSLY VERY PROVOCATIVE, THE
DATA I SHOW YOU IS NOT COMPLETE
AND I THINK IT'S VERY
INTERESTING AND AGAIN, YOU KNOWS
THIS QUESTION, YOU KNOW HOW CAN
YOU USE HUMAN DATA OR MOUSE DATA
TO GET THE MOST BEING ABOUT IT.
SO RIGHT NOW THIS IS ONGOING IN
THE CLINICAL CENTER AND I WILL
PUT MY HEAD ON AS A MEDICAL
ONCOLOGIST.
SO AS A MEDICAL ONCOLOGIST, WE
USED ANTICDLA4, AND ANTIPDL1,
AND COLLEAGUES THAT USUALLY
WORKS THEY HAVE 1 DRUG THAT
WORKS, THEY HAVE A SECOND DRUG
WORKS AND THEN THEY ASK THE
SCIENTIFIC QUESTION, WHAT HAPPEN
FIST YOU COMBINE THEM?
SO I DID THE SAME THING.
I USED ALL THE DRUGS HERE, WE
PERFOR THE PURPOSED READ OUTS
AND BIOPSIES, BLOOD SAMPLES,
LOOKED AT TAIJING, ET CETERA.
AND TOLD YOU WE HAD DONE STUDIES
BEFORE WHERE WE ONLY USED THE
SAME COMBINATION BUT NO
ANTIVEGFOR OR THE SAME
COMBINATION WHERE WE DIDN'T HAVE
ANTIPUNISHING DL1.
OKAY, SO THIS IS SUPER EXCITING.
BUT I SEE THIS WE ONLY TREATED 7
PATIENTS, AND YOU CAN SEE THE
OUTCOME AND THIS AGAIN SOMETHING
YOU HAVE TO TAKE WITH A GRAIN OF
SALT BECAUSE WE CANNOT COMPARE
BETWEEN TRIALS AND THERE ARE
DIFFERENCE IN DOSING, ET CETERA,
SO YOU KNOW THIS IS ALL-I TAKE
ALL THOSE CRITICISM AND THESE
ARE VERY SMALL PATIENT NUMBERS,
BUT THE DATA IS QUITE
SURPRISING.
YOU CAN SEE THAT ONCE WE USE THE
[INDISCERNIBLE] IN RED, ON THE
BOTTOM YOU SEE THE PROGRESS FREE
SURVIVAL, IT'S SIGNIFICANTLY
LONGER THAN BEFORE, BELIEVE ME,
THE PATIENTS WE SEE HERE HAVEN'T
CHANGED MUCH IN THE PAST 6 OR
8IERS AND THIS ALSO LEADS TO A
DIFFERENCE IN SURVIVAL AS CAN
YOU SEE ON TOP WITH JUST A VERY
FEW PATIENTS.
NOW IF IT WAS ONLY THIS, I WOULD
NOT BE VERY EXCITED BECAUSE I
WOULD SAY IT'S PROBABLY PATIENT
SELECTION EMPLOY YOU KNOW, 7
PATIENTS, IT'S REALLY NOTHING,
BUT THERE'S MORE.
THESE PEASHTS DEVELOP
SIGNIFICANT IMMUNE RELATED
ADVERSE EVENTS.
SO CAN YOU SEE HERE ON THE
RIGHT, THE 7 PATIENTS.
YOU CAN SEE THE DAY WHEN THESE
IMMUNE RELATED ADVERSE EVENTS
OCCURRED IS YOU CAN SEE THE
GRADE.
AND I WILL LIKE TO POINT YOUR
ATTENTION TO THE ADVERSE EVENTS
THAT TARTS WITH AN M.
SUCH AS MYOCYTEIS, MYASTHENIA,
MY O CARDITEIS, ATRIAL
FIBRILLATION, THESE ARE NOT THE
TYPES OF EVENTS WE USUALLY FIND
IN OUR PATIENTS.
AND I KNOW THIS' A LOT OF LAB
PEOPLE IN HERE SO I THOUGHT I
WOULD GO DEEPER AND SHOW YOU A
SLIDE FROM 1 OF THE
PRESENTATIONS THAT I GOT FROM
OUR COLLEAGUES FROM NEUROLOGY
THAT DESCRIBED THE HOSPITAL
COURSE OF 1 OF THESE PEASHTS AND
I JUST HIGHLIGHT A FEW OF THESE
YOU SEE INDUCED--NEW CHECK POINT
INHIBITOR--SO THESE ARE HEART
ENZYMES, YOU SEE MY O CARDITEIS,
YOU SEE WEAKNESS, THE PATIENT
WAS UNAWARE OF THE EXPENTINE
REGIMEN OF HIS SWALLOWING
IMPAIRMENT, EYE MOVEMENT GOT
BETTER, THE VOICE WAS SO HORSE
HE WAS NOW USING A VOICE
AMPLIFIER SO OUR PATIENT
COULDN'T REALLY SPEAK ANYMORE,
AND THE PRIMARY TEAM SO THAT'S
US, WE EVEN CONSIDERED A
PUNISHING EC FOR THAT PATIENT
ISSUES THAT'S FOR THE PEOPLE WHO
DON'T DO THE WORK AT THE TIME,
THEY PUT A TUBE INTO THE
PATIENT.
SO WE SEE SIGNIFICANT RELATED
ADVERSE EVENTS IN THESE
PATIENTS.
THIS CAUGHT OUR ATTENTION AND
I'M NOT GOING TO TELL YOU MORE
ABOUT THE CLINICAL PART BUT WHAT
I THINK--WHAT I THOUGHT AT THAT
POINT WAS, THERE MUST BE A VERY,
VERY STRONG IMMUNE RESPONSE.
SO THIS IS NOW THE--OUTLINE FOR
THE CORRELATIVE STUDIES FOR THIS
PROJECT AND DON'T WORRY, I WILL
NOT SHOW YOU ALL OF THIS BUT WE
HAD TUMOR BIOPSIES AND I WE
TRANSLATED ALL THIS INTO A
PHOSPHORYLATED MODEL TO VERIFY
AGAIN SIMILAR TO WHAT I'VE SHOWN
YOU BEFORE, YOU KNOW WHAT THE
UNDERLYING MECHANISM IS.
LET'S START ANALYZING THE
T-CELLS, WE SAW AS EXPECTED A
VERY STRONG KD8 T-CELL RESPONSE
APPROXIMATE WHEN I SAY DOUBLE
VERSUS TRIPLE ISSUE, WE
COLLECT--COMPARE SORRY, FROM THE
PASHTS THAT ONLY RECEIVE CTLA-4
AND PDL1 WITH THE 1S THAT
RECEIVE THE DOUBLET WITH THE
ANTIBGF.
BUT WHAT IS IT DOING?
WHAT IS CONSIDERED THE MOST
STRONG IMMUNE CHECK POINT
INHIBITOR, BUT APPARENTLY CD8
T-CELLS ARE WAY MORE ACTIVATED.
AND THEN QUITE SURPRISINGLY
THERE WAS AN INCREASE IN
REGULATORY T-CELLS.
WHY REGULATORY T-CELLS.
REGULATORY T-CELLS WOULD
SUPPRESS AN IMMUNE RESPONSE AND
I JUST TOLD YOU THESE PATIENT
HIS A GOOD OUTCOME AND THEY
DEVELOP CD8 IMMUNE RESPONSES AND
WE KNOW THESE REGULATORY T-CELLS
CAN REPRESS RESPONSES THROUGH A
DIFFERENT MECHANISM THROUGH
APCs AND SO WE KNOT REALLY
INTERESTED IN THIS.
THE OTHER FINDING WE MADE WAS
THE FOLLOWING.
WE HAD SERUM SAMPLES AND WE
COMPARE THEM WITH THE 1S THAT
HAVE THE TREATMENT, ASK WE SAW
THEY MADE A LOT OF IL12.
THIS IS THE FIRST COLUMN .
IF WE TREAT THESE PATIENTS WITH
ANTIBGF, SO WHAT IS THE
UNDERLYING MECHANISM?
SO WE WENT AHEAD AND TREATED OUR
MICE, WE TREATED THEM THE SAME
WAY, GAVE THEM THE COMBINATION,
AND INDEED WE WERE ABLE TO
REPEAT THIS.
WAWHAT WE HAD SEEN IN OUR
PATIENTS, WE SAW AN INCREASE OF
REGULATORY T-CELLS.
NOW WE WERE ABLE TO LOOK AT
THESE T-REGS IN MORE DETAIL, WE
SHOW THAT THEY MAKE A LOT OF
INTERFERON-GAMMA.
THEY TART TO UPREGULATE THE
TRANSCRIPTION FACTOR, AND THEY
DOWN REGULATE ANOTHER 1 HERE.
SO THIS SUGGESTS ALREADY THERE
MUST BE SOMETHING HAPPENING TO
THESE T-REGS THAT FIRST OF ALL
THEY START BECOMING LIKE AN
EFFECTOR CELL BECAUSE THEY MAKE
INTERFERON-GAMMA AND THE
PHENOTYPE IS CHANGING.
WHAT ABOUT THE IL12 THEY ARE
ACTUALLY LESS IMMUNOSUPPRESSIVE,
THEY MAKE MORE INTERFERON-GAMMA
AND SOMEHOW THEY ALSO LOSE
SUPPRESSOR FUNCTION EMPLOY NOW
WHAT ABOUT IL12.
IF WE USE ANTIIL12, WE ACTUALLY
LOSE THE EFFECT THAT WE GAIN BY
ADDING ANTIBGF, SO THERE SEEMS
TO BE ANOTHER CONNECTION THAT WE
HAD BASICEE PREDICTED BASED ON
OUR HUMAN DATA.
YOU CAN SEE FIRST A TUMOR SIZES
AND THE TUMOR WEIGHT AND THEN
AGAIN OUR CD8 T-CELLS ALL OF A
SUDDEN NO LONGER MAKE IL12,
SORRY THE T-REGS MAKE NO LONGER
LARGE AMOUNTS OF
INTERFERON-GAMMA THAT WE HAD
SEEN BEFORE AND THEY ALSO DON'T
CHANGE ANYMORE, DEDECREASE IS
NOT AS SIGNIFICANT AND WE DON'T
SEE A CHANGE IN T-BET.
SO IT HAD BEEN SUGGESTED BY
[INDISCERNIBLE]'S GROUP THE
NORMAL TISSUING REGS WE KNOW
EXPRESS A LOT OF Foxp3 AND ARE
IMMUNOSUPPRESSIVE AND BUT THEY
CAN CHANGE INTO FRAGILE AND
UNSTABLE TREGS, THEY LOSE
EXPRESSION OF Foxp3, THEY TART
TO RELEASE INTERFERON-GAMMA AND
THEY CHANGE EXPRESSION AND
EXPRESS Foxp3 AND START TO
BECOME LESS IMMUNOSUPPRESSIVE.
SO IF THIS IS TRUE WE SHOULD SEE
THIS IN OUR PATES.
SO NOW WE WENT BACK TO THE
PATIENTS, WE AND WHAT I'M DOING
HERE NOW IS THIS IS ALMOST THE
LAST SLIDE, WE LOOKED AT THE
EFFECT ISSUES WE SPECIFICALLY
ZOOM IN ON THE TREGS AND PREAND
POST TREATMENT ASK WE SEE THAT
THESE TREGS DON'T CLUSTER INTO 1
GROUP.
CAN YOU SEE THERE IS THE--THERE
ARE DIFFERENT SUBSES, SOME FOR
INSTANCE IF YOU LOOK AT CTLA-4,
THEY ARE THE STONES, THE SAME
APPLIES TO STAT 1 OR YOU KNOW
MANY OF THE OTHER MOLECULES THAT
YOU WANT TO LOOK AT AND IF WE DO
THIS, LOOK IN MORE DETAIL, WE
CAN ACTUALLY E DENTIFY HERE, THE
DIFFERENT T-CELL SUBSETS, THE
NTEGGIC--STRATEGIC REGS WHICH
ARE THE NATURAL TREGS WE BELIEVE
ARE VERY IMMUNOSUPPRESSIVE BUT
ALSO THE FRAGILE T-REGS AND THE
UTREGS, THE UNSTABLE TREGS, THIS
IS NOW THE ANALYSIS FROM ALL
THESE PATIENTS BEFORE AND AFTER
AND INTERESTINGLY IF WE COMPARE
PRE VERSUS POST, YOU CAN SEE
THAT THE RED CLUSTER GETS
SMALLER EMPLOY THESE ARE THE
NTREGS, THE UTREGS, THE GREEN 1
AS WELL AS FRAGILE TREGS, THESE
ARE THE 1S THAT MAKE MORE
INTERFERON-GAMMA AND THAT LOSES
SUPPRESSOR FUNCTION INCREASE IN
THESE PATIENTS.
AND THEN AGAIN, I KNOW YOU ARE
ALL DOING SINGLE CELL ANALYSIS
AND YOU DO THIS ALL THE TIME, SO
LINEAGE COMMITMENT AND WOE SEE
THAT INDEED, IN THE PATIENTS
THESE CELLS--THESE FRAGILE TREGS
AS WELL AS UTREGS CAME OUT OF
THE NTREG.
SO LET ME SUMMARIZE THE LAST
PART.
THE COMBINATION OF ANTIBGF, AND
THIS EVOKED VERY IMPORTANT
ANTITUMOR IMMUNITY.
I'VE SHOWN YOU THAT THE ADDITION
OF THE VEGF INCREASED SYSTEMIC
IERK L12 AND THIS LEADS TO AN
IL12 DEPENDENT CHANGE IN TREGS
WITH AN INCREASE IN FRAGILE AND
UNSTABLE TREGS.
SO WITH THIS, I WILL LIKE TO
CLOSE AND NEED TO THANK THE
GROUP, BUT BEFORE I THANK THE
GROUP THAT'S PRESENT, I THINK
IT'S IMPORTANT TO SPEBD A BRIEF
MOMENT TO THANK ALL THE PEOPLE
THAT GENERATE THIS DATA AND
ALREADY LEFT MY LAB AS WELL AS
THE TREMENDOUS CLINICAL SUPPORT,
WITHOUT THEM IT WOULD BE
IMPOSSIBLE TO DO THESE STUDIES
BOTHOT CLINICAL SIDE AS WELL AS
ON THE SIDE THAT WE NEED, YOU
KNOW ADMIN TRAITIVE SIDE TO DO
TRIAL SUPPORT.
AND THEN OBVIOUSLY, FOR THE
SCIENCE PART THE MANY, MANY
DIFFERENT NIH COLLABORATORS THAT
HELP WITH DIFFERENT ASPECTS OF
MICROBIOME AND OBVIOUSLY LIVER
CANCER AND IMAGING, I MENTIONED,
ET CETERA, THE VARIOUS CORE
FACILITIES, BIOINFORMATTIC
RESOURCES THAT HELP, EXTRAMURAL
COLLABORATORS AND OBVIOUSLY LAST
BUT NOT LEAST ALSO ADDITIONAL
FUNDING THAT I WAS ABLE TO
COLLECT THAT MADE THESE STUDIES
POSSIBLE AND WITH THIS, I AM
HAPPY TO TAKE ANY QUESTIONS.
KD--SALLY ASH PLAWZ
>> OVER HERE EMPLOY BEAUTIFUL
STORY.
ONE QUESTION, COMMENT ABOUT THE
LAST PART OF THE STORY WITH THE
ANTIVEGF.
SO AS YOU KNOW VEGF ITSELF
INDUCES AT THAT TIME 3 AND
ANOTHER CYTOKINE THAT INDUCES
STAT 3 AND IL10 WHICH KNOWN TO
INHIBIT PRODUCTION OF IL12.
SO 1 MIGHT EMBASSY THAT VEGF
COULD INHIBIT IL12 AND THEREFORE
ANTIVEGF MIGHT PROMOTE MORE IL12
PRODUCTION WHICH WOULD INDUCE
T-BET AND INTERFERON-GAMMA
PRODUCTION AND SO FORTH BY THE
TREGS.
SO CAN YOU LOOK TO SEE WHETHER
THE PHOSPHORYLATION OF STAT 3
PLAYS A ROLE IN THIS PROCESS
THAT YOU DESCRIBED?
>> SO HAD IS THE NICE THING
ABOUT THE MOUSE FORM AND WHY I
LIKE THESE TYPE OF STUDIES,
REMEMBER WE ONLY HAD 7 PATIENTS
AND YES, IN MICE WE CAN DO
EVERYTHING, YOU KNOW WE CAN LOOK
AT STAT 3 BECAUSE THESE
EXPERIMENTS ARE NOW RELATIVELY
EASY TO DO.
SO AS I MENTIONED, AS YOU KNOW
BECAUSE WE ACTUALLY HAVE A
SHARED LAB MEETING FOR EVERYBODY
ELSE, SO YOU KNOW JAY ACTUALLY
KNOWS ALL THIS DATA, OBVIOUSLY,
YOU KNOW WE CAN DECIPHER THIS IN
MORE DETAIL TO SEE WITH THE IL10
PLAYS A ROLE IN THIS.
>> OKAY, THANKS.
>> HI DR. GRETEN, I VERY MUCH
ENJOYED YOUR TALK AS I DID AS MY
TIME IN YOUR CLINIC AS AA
TRAINEE.
MY QUESTION HAS TO DO WITH
INNATE CELL INHIBITION GROWTH.
DO THE MAIT CELLS POSSESS THE
CAPACITY FOR METASTASIS AND ALSO
DO THEY HAVE THE SAME WITH THE
SCLEROTICS LIVER WITH THE
COLORECTAL MEDS?
THANK YOU.
>> O THAT IS OBVIOUSLY A VERY
GOOD QUESTION AND PART OF THE
MATTER OR ABILITY OR YOB IS TO
PROVIDE YOUR FELLOWS WITH GOOD
PROJECTS.
[LAUGHTER]
>> THANK YOU.
>> SO THIS POST DOC LEFT AND
STARTED HIS OWN LAB, HE'S GOING
TO STUDY NOT WHAT YOU PROPOSE
BUT HE WILL IS ITED THIS IN THE
CONTEXT OF LIVER DEC AND FATTY
LIVER DISEASE WHICH KIND OF WHAT
YOU SUGGEST, AND YOU KNOW I LET
THEM DO THE STUDIES.
IF YOU WANTED TO STUDY THIS IN
THE CO KNOW SEPTORS, WE BEING
POTENTIALLY DO THIS, TOO.
>> THANK YOU.
>> ALWAYS A HUGE PLEASURE AND
I'M EYE HUGE FAN OF TIM'S WORK
SO IT'S GREAT TO BE HERE.
THANK YOU SO MUCH.
I HAVE A QUESTION WHICH MAY BE 1
OF IGNORANCE BUT IN THE CONTEXT
OF THE MAIT CELLS AND THINKING
ABOUT THE MICROBI OTA DO YOU
KNOW HOW RIBOFLAVORIN LEVELS
PLAY A ROLE IN ALL THIS AND
COULD YOU MANIPULATE THAT?
RIGHT?
>>
>> YEAH, NO, I KNOW.
SO FIRST OF ALL I DON'T KNOW.
THAT'S NUMBER 1.
WE KNOW THAT, YOU KNOW FROM
JASMINE'S WORK THAT YOU NEED THE
GUT MICROBIOME TO HAVE THE
MAITs, AND YOU KNOW THIS, BUT
THE AUDIENCE PROBABLY DOESN'T
KNOW THIS IS IT'S VERY DIFFICULT
TO STUDY MAITs IN MICE,
BECAUSE THEY DON'T HAVE A LOT OF
NKT CELLS, IT'S HUMANS THAT HAVE
THE MAITs, YOU KNOW I KIND OF
DISMISS THAD PART SO IT'S REALLY
DIFFICULT TO DO THOSE STUDIES
WHY JUST BECAUSE THERE'S SUCH
FEW CELLS.
N[INAUDIBLE RESPONSE FROM
AUDIENCE ]
>> WE HAVEN'T LOOKEDDA THE THIS
BUT WHAT WE'RE BASICALLY--WE
DON'T KNOW.
BUT WHAT WE'RE TRYING TO DO NOW
IS, YOU KNOW WE ARE TRYING TO
FIND ALTERNATE WAYS TO INDUCE
MAITs AND THEN RATHER
INDEPENDENT OF [INDISCERNIBLE]
TO USE THEM AS A TOOL TO INDUCE
ANTITUMOR IMMUNITY.
>> LET ME ASK YOU 1 QUESTION,
STEMMED FROM THE LAST QUESTION
AS AN EXTENSION, MORE AS A
GLOBAL.
SO YOU FIND PLATELETS FOR
SIGNALING, YOU ALSO FIND THE
MAITs FOR SIGNALING AND IN THE
MICROBIOME AS WELL, BUT IT STEMS
FROM DISEASE AND MULTIPLE
IMMUNOLOGYS THAT CONTRIBUTE AND
HOW DO YOU SEE THAT IF IT'S A
DIFFERENT CONNECTION THAT HELP
TO IMPROVE THE TREATMENT,
WHETHER THEY'RE FUNCTIONAL AT
LENGTH NOW.
>> WELL OBVIOUSLY ANY OF THESE
SHOW THE MECHANISM OF SPECIFIC
SECTOR AND IN THE END, YES, THE
QUESTION IS YOU KNOW HOW YOU--IN
THE END THE PERSON WHO WINS IS
THE 1 WHO TREATS THE PATIENT AND
THE PATIENT RESPONDS.
THERE'S NO QUESTION.
BUT IN THE END AUTOWHAT'S MOST
IMPORTANT AND SIGNIFICANT.
AND THE IT'S A DIFFICULT
QUESTION AND A LONG QUESTION BUT
I DO HAVE IDEAS ABOUT THIS.
IT'S NOT THAT I WANT TO RUN AWAY
FROM THIS, I THINK A LOT OF THE
WORK THAT'S GOING ON RIGHT NOW
IS VERY T-CELL CENTRIC AND
THAT'S BECAUSE PEOPLE LIKE
T-CELLS, WE HAVE STUDIED T-CELLS
FOR A LONG TIME AND WE CHECK
POINT INHIBITORS, SO THAT'S WHY
PEOPLE STUDY THEM CAN WE TRY TO
STUDY THEM AS LITTLE AS POSSIBLE
BECAUSE THERE'S MANY OTHER VERY
IMPORTANT IMMUNE CELLS THAT ARE
DISMISSED AND THAT'S WHAT I TRY
TO SHOW YOU HERE IN THE END, SO
IF YOU ARE REALLY ASKING ME FROM
A BIG PICTURE, THE MYELOID
COMPARTMENT, I SHOWED YOU THE
TUMOR ASSOCIATED MACROPHAGES CHU
ONLY FIND IN THE SURROUNDING
TISSUE UPREGULATING PDL1, OUR
CERTAINTY IN CELL POPULATION
WHICH IS OF GREAT INTEREST,
THEY'RE VERY IMMUNOSUPPRESSIVE
AND THE SAME AMRIS TO REGULATORY
T-CELLS, WE HAVE ANOTHER STUDY
WHERE WE LACK THEA REGULATORY
T-CELLS WE KNOW THESE ARE MANY,
MANY YORES, WE ARE JUST NOT ABLE
TO SPECIFICALLY TARGET AND
ELIMINATE THEM.
WE DO THIS BY EXITING AND USING
ANTIVEGF, WE HAVE DRAMATIC
CENTER FOR EXCELLENCE ON AGINGS
BUT I THINK WHAT THE FIELD
REALLY HAS TO LEARN AND DO A
LITTLE BIT IS TRY TO FIND MORE
SIGNIFICANT CHANGES, RATHER THAN
THIS INCREMENTAL CHANGES USING
WHAT I DID MYSELF, ADDING PDL1
AND CTLA-4, GO FOR THE BIG
CHANGE.
NOKAY, WELL THIS IS A PERFECT
WRAP UP OF WHAT IS A FANTASTIC
GRAND ROUND.
THANK YOU VERY MUCH, TIM.