GOOD MORNING, EVERYONE.
MY NAME IS STEPHANIE GOFF AND I
WAS CHAIR OF THE PLANNING
COMMITTEE FOR THIS CONFERENCE SO
ON BEHALF OF DR. ROSENBERG AND
MYSELF, THANK YOU ALL FOR
ATTENDING.
I'D LIKE TO ASK DR. JAMES GULLE
TO GIVE US SOME OPENING REMARKS,
CO-DIRECTOR OF THE CENTER FOR
IMMUNOONCOLOGY, THE CLINICAL
DIRECTOR HERE AT THE NCI AND
ACTING CO-CHAIR OF THE CENTER
FOR CANCER RESEARCH.
PLEASE WELCOME DR. GULLE.
[APPLAUSE]
>> THANK YOU SO MUCH.
IT'S REALLY A DELIGHT TO BE HERE
TODAY AND ON BEHALF OF THE
INCREDIBLE INVESTIGATORS DOING
OUTSTANDING RESEARCH HERE AT THE
CENTER FOR CANCER RESEARCH, I'D
LIKE TO BOTH WELCOME YOU AND I'M
DELIGHTED AND HONORED TO KICK
OFF THIS SYMPOSIUM ON THE PAST,
PRESENT AND FUTURE OF CELLULAR
IMMUNOTHERAPY.
SO AS SOMEONE WHO EARLY IN MY
CAREER REALLY DRANK THE KOOL-AID
OF IMMUNOTHERAPY, IT'S BEEN SUCH
AN INCREDIBLE RIDE TO SEE US
GOING FROM THE APPROVAL OF IL-2
AS YOU'RE GOING TO HEAR ABOUT IN
A LITTLE BIT FROM DR. LOTZE
THROUGH WHERE WE ARE TODAY, WITH
A MULTIBILLION DOLLAR INDUSTRY
AIMED AT HELPING PATIENTS, BUT
MORE IMPORTANTLY, ALL THE LIVES
IMPACTED, AND ALL THE PATIENTS
CURED.
AS ONE OF THE PRIMARY DRIVERS OF
THIS WORK WAS RIGHT HERE IN THE
INTRAMURAL PROGRAM IN THE NIH.
AND THIS MORNING, WE ARE MEETING
HERE IN THE AUDITORIUM OF THE
NIH CLINICAL CENTER, AND I JUST
WANT TO SAY THAT THE NIH
CLINICAL CENTER IS THE LARGEST
CLINICAL RESEARCH CENTER IN THE
WORLD, AND THE NCI'S CENTER FOR
CANCER RESEARCH IS ONE OF ITS
MAJOR USERS.
THE NIH CLINICAL CENTER IS NOT A
REGULAR HOSPITAL, IT'S REALLY
DEDICATED TO PATIENT RESEARCH
AND INTENSIVE PATIENT RESEARCH,
AND ALL OF THE PATIENT TREATED
HERE ARE TREATED ON CLINICAL
TRIALS.
THE NCI INTRAMURAL CLINICAL
PROGRAM IS A RESOURCE FOR BOTH
THE BASIC SCIENCE AND CLINICAL
SCIENTISTS AND REALLY WHAT WE
EXCEL IN IS TRANSLATING THESE
BASIC DISCOVERIES INTO FIRST IN
HUMAN CLINICAL TRIALS.
I WANT TO JUST SHARE A LITTLE
BIT MORE ABOUT THE CENTER FOR
CANCER RESEARCH.
WE HAVE CENTRAL FUNDING WHICH
ALLOWS US TO FLEXIBLY SPEED
THINGS INTO THE CLINIC.
IT ALLOWS US THE INDEPENDENCE TO
ALLOW HIGH RISK RESEARCH.
WE ALSO HAVE CLINICAL RESOURCES
THAT WE DON'T BILL PATIENTS OR
INSURANCE.
WE WILL PAY FOR PATIENTS TO
TRAVEL FROM ANYWHERE IN THE
UNITED STATES TO COME HERE ON
CLINICAL TRIALS, AND THIS REALLY
ALSO AFFORDS EXCELLENT
OPPORTUNITIES TO TRAIN PHYSICIAN
SCIENTISTS BECAUSE WE DON'T HAVE
AN RVU-BASED SYSTEM.
IN ADDITION, OUR COLLABORATIVE
ATMOSPHERE REALLY ENCOURAGES
TEAM SCIENCE AND PARTNERING.
MORE ON THAT IN A LITTLE BIT.
SO THIS IS A SNAPSHOT OF SOME OF
OUR METRICS ON OUR CLINICAL
RESEARCH ACTIVITY.
WE HAVE ABOUT 1700 NEW PATIENT A
YEAR HERE, AND IN ADDITION TO
OUR NATURAL HISTORY STUDIES AND
NON-TREATMENT STUDY, WE HAVE
ABOUT A THOUSAND PATIENTS THAT
ARE ACCRUED TO INTERVENTIONAL
STUDIES EACH YEAR HERE.
TYPICALLY THESE ARE IN PHASE ONE
OR PHASE TWO FIRST IN HUMAN
STUDIES.
IN TERMS OF CELL THERAPIES,
THERE ARE CURRENTLY 29 OPEN CELL
THERAPY STUDIES IN HERE, AND
WE'RE TALKING ABOUT CAR T, TCR
Ts, TILs, BUT ALSO NK CELLS,
MYELOID CELLS AND MONOCYTES.
THESE CELL THERAPY STUDIES ARE
ONGOING IN SIX DIFFERENT
BRANCHES HERE, WHICH IS ABOUT A
THIRD OF THE GROUPS DOING
CLINICAL TRIALS.
SO I'M PROUD OF THIS SLIDE HERE.
THIS SLIDE OUTLINES SOME OF THE
RESEARCH THAT HAS LED TO NEW
TREATMENTS BEING APPROVED FOR
PATIENTS AS A TESTAMENT TO SOME
OF THE CLINICAL SCIENCE GOING ON
HERE.
YOU CAN SEE THAT OVER THE PAST
SEVEN YEARS, WE'VE HAD 12 NEW
THERAPIES THAT WERE
FDA-APPROVED, AND WHAT I'VE
OUTLINED IN RED HERE ARE THE
IMMUNOTHERAPIES THAT HAVE BEEN
APPROVED BY THE FDA.
IN ADDITION, THERE'S BEEN
MULTIPLE BREAKTHROUGH THERAPY
AND ORPHAN DRUG DESIGNATION
BASED ON WORK GOING ON HERE.
BUT I'D JUST LIKE TO HIGHLIGHT
THESE TWO CELL THERAPIES BASED
ON WORK THAT CAME OUT OF THE
SURGERY BRANCH.
AND YOU'RE GOING TO HEAR MORE
ABOUT THESE, I'M SURE, LATER ON
TODAY.
SO WE'VE COME FROM CYTOKINES TO
IMMUNE CHECKPOINT INHIBITORS AND
CELL THERAPY, BUT WHERE ARE WE
GOING TO GO NEXT?
AND SO THIS IS A PAPER THAT WAS
PUT TOGETHER AUTHORED BY EMENS
AND COLLEAGUES THAT OUTLINES
SOME OF THE CHALLENGES AND
OPPORTUNITIES IN THE FIELD.
I WANTED TO BRING UP SOME OF THE
THINGS THAT WAS BROUGHT OUT IN
THIS PAPER AND REALLY ONE OF THE
POINTS THAT WAS BROUGHT OUT WAS
THE POTENTIAL FOR PARTNERSHIPS.
HERE.
SO PARTNERSHIPS BETWEEN ACADEMIC
INSTITUTIONS, PARTNERSHIPS WITH
COMPLEMENTARY SCIENCE
DISCIPLINES AND PARTNERSHIPS
BETWEEN PHARMA AND BIOTECH.
ON THE LATTER, I'D JUST LIKE TO
MENTION THAT ABOUT 70% OF NEW
DRUGS COME FROM BIOTECH.
SO BASED IN PART ON THAT, THE
SOCIETY FOR IMMUNOTHERAPY OF
CANCER OUTLINED THEIR
BIOTECHNICIANTIVE, WHICH REALLY
WHAT OUR GOAL IS, IS TO GET 100
NEW APPROVALS WITHIN THE NEXT
DECADE.
AND SO FAR THIS YEAR, YOU CAN
SEE THERE'VE BEEN FOUR
APPROVALS, AND THESE ARE
HIGHLIGHTED HERE BELOW, AND YOU
CAN SEE THAT TWO OF THESE ARE IN
CELL THERAPIES.
SO ONCE AGAIN, I'D LIKE TO
WELCOME EACH AND EVERY ONE OF
YOU TO THIS SYMPOSIUM, AND I'D
LIKE TO THANK MY FRIEND AND
COLLEAGUE, DR. STEVE ROSENBERG,
FOR HIS TENACIOUS BELIEF IN
LIVING THERAPIES THAT HAVE
BROUGHT US ALL HERE.
THANK YOU.
[APPLAUSE]
>> HELLO, EVERYONE.
THANK YOU SO MUCH FOR COMING.
I'M NICK CLEMEN, I'LL BE
MODERATING THE FIRST SESSION.
THE TALKS ARE ONLY 15 MINUTES
LONG AND WE'RE TRYING TO KEEP ON
SCHEDULE SO WE'LL HOLD THE
QUESTIONS FOR THE END.
SO WITHOUT FURTHER ADO, FIRST UP
IS DR. MICHAEL LOTZE FROM THE
UNIVERSITY OF PITTSBURGH.
>> WELL, FIRST OF ALL, IT'S AN
UNUSUAL PLEASURE AND HONOR TO BE
ABLE TO BE HERE AND GIVE THE
FIRST TALK.
I WOULD LIKE TO SAY THAT ALL
RHODES LEAD TO IL-2 AND LED FROM
IL-2 FOR CELL THERAPY.
BUT I'M BIASED.
AND THANKS, JAMES GULLEY, FOR
THAT NICE INTRODUCTION.
I THINK WE STILL HAVE A LONG WAY
TO GO TO GET 100 APPROVALS, BUT
WE'RE ON OUR WAY.
SO I CALL THIS A PROVOCATIVE
TITLE, THE FIRST EFFECTIVE
CHECKPOINT THERAPY WAS IL-2, IN
PART BECAUSE AT THE TIME WHEN WE
DID IT, WE DIDN'T KNOW THAT WE
HAD TO GET OVER ALL THOSE
GLUTTONOUST REGS THAT ATE OUR
IL-2, IT MEANT WE HAD TO GO TO
HIGH DOSE.
THE UNIVERSITY OF PITTSBURGH IS
NOW PUTTING UP, FINALLY, AFTER
35 YEARS THERE, A CELL THERAPY
FACILITY.
I DO WANT TO PAUSE A MINUTE AND
SAY ONE OF THE GRADUATES OF OUR
PROGRAM HERE AT THE SURGERY
BRANCH OF THE NCI, JEFF WEBBER,
SADLY PASSED JUST A FEW MONTHS
AGO, AND HE MOVED FROM BEING ONE
OF OUR FELLOWS INTO A POSITION
ON THE SENIOR STAFF, AND THEN AN
ILLUSTRIOUS CAREER THAT TOOK HIM
THROUGH MOFFITT AT ONE POINT AND
THEN ALSO TO NYU, WHERE HE
FINISHED HIS CAREER.
I SHOULD SAY THAT I CAME TO THE
BRANCH IN 1978.
THE LAST MILLENNIUM, THE LAST
CENTURY.
AND TWO PEOPLE WHO TAUGHT ME HOW
TO DO RESEARCH WERE ELANA RNA IN
WHO STARAND SUSIE SCHWARTZ.
I HAD MY OWN SMALL GROUP IN THE
BRANCH.
THIS IS JOSH REUBEN, HE JUST
RETIRED AS A SURGEON.
MARY, ALL THE WAY UP TO THE TOP
OF THE NCI.
DAVE JABLINS WHO RAN A BAGEL
BUSINESS IN SAN FRANCISCO AS
WELL AS BEING HEAD OF THORACIC
SURGERY AT UCSF.
AND YUTAKA KAWAKAMI, WHO I'M
DELIGHTED TO SEE HERE AFTER SO
MANY YEARS.
AND THIS IS A GROUP THAT I WAS
ABLE TO BUILT AT UNIVERSITY IN
PITTSBURGH WHEN I LEFT IN 1990
BUT IT WAS ALL PREDICATED ON
THIS EARLY GROUP THAT STEVE PUT
TOGETHER.
THIS IS A PICTURE FROM HIS BOOK
IN 1982, MANY OF YOU WILL
RECOGNIZE SOME OF THE PEOPLE
THERE, BUT IT INCLUDED FRED
CHANG, LIZ GRIM, STEVE
EDINGHAUSEN AND JIM WHO YOU'LL
HEAR FROM LATER.
I HAVE SOME DISCLOSURES, HERE
THEY ARE.
THIS IS A 50 YEAR CELEBRATION
FOR STEVE ROSENBERG BUT AN
ORGANIZATION THAT I'VE BEEN
ASSOCIATED WITH FOR MUCH OF MY
CAREER IS GOING TO ITS 39TH
MEETING IN HOUSTON, SITC, I
ENCOURAGE YOU ALL TO CONSIDER
IT, AND ALSO COME BACK WHEN IT'S
BACK IN WASHINGTON FOR THE 40TH
ANNIVERSARY.
I SHOULD SAY THAT TUMOR
IMMUNOLOGY IS REALLY COMPLICATED
AND CONFUSING.
THIS IS A PICTURE FROM GLEN
DRANOFF FROM 20 YEARS AGO WHERE
YOU SEE ALL THIS BLISTERING
ARRAY OF CELLS, CYTOKINES AND
CHEMOKINES AND IT'S REALLY
CONFUSING BUT IF YOU REALLY LOOK
AT IT, YOU EITHER GET ENDOGENOUS
IL-2 BEING MADE OR GIVE
EXOGENOUS IL-2 AS THE ENDOGENOUS
COMING FROM SIGNAL 1 AND SIGNAL
2, OR AS I DID WHEN I WAS AT
NEREX USE A B INHIBITOR BUT THE
REAL PROBLEM WAS WE DIDN'T KNOW
ABOUT IT. REGS AND GIVING
EXOGENOUS IL-2 WAS ALWAYS A
DICEY PROPOSITION AS WE LEARNED.
SO THE FIRST 40 YEARS OF CANCER
IMMUNOTHERAPY FROM MY STANDPOINT
IS WE ACTIVATED WITH WHAT WAS
THEN CALLED T-CELL GROWTH
FACTOR, SUBSEQUENTLY CALLED
IL-2, LYMPHOKINE ACTIVATED
KILLER CELLS.
I DID THAT AS A CLINICAL
ASSOCIATE WORKING WITH STEVE IN
1980.
WE STARTED OUR IL-2 CLINICAL
TRIALS IN THE MID 80s.
SUZANNE TOOK OVER MY LAB WHEN I
LEFT THE NIH, HAD AN ILLUSTRIOUS
CAREER HERE, ADVANCING TUMOR
INFILTRATING LYMPHOCYTES, SHOWN
IN RED, SUBSEQUENTLY BECAME A
GREAT CHAMPION FOR PD-1 AND
PDL1.
YUTAKA KAWAKAMI DID WONDROUS
STUDIES EVALUATING ANOTHER
T-CELL GROWTH FACTOR, IL-4 BUT
THEN WENT ON TO CLONE MART1 AND
M 1 ANDMELAN A.
DENDRITIC CELLS BEING GIVEN TO
PATIENTS, WENT ON TO DEFINE THE
ROLE OF THE MICROBIOME AND
IMMUNOGENIC CELL DEATH, AND THEN
DAL WIN TONG.
THAT'S A 40 YEAR HISTORY.
BUT ALONG THE WAY AND THANKFULLY
FOR ALL OF US, THE CHECKPOINTS
CAME ALONG IN 2010 AND 2014.
SO IF YOU LOOK AT IT, WE HAD T
CELLS AND T-CELL GROWTH FACTOR,
IL-2 IN THE BEGINNING.
A LONG PERIOD TESTING TUMOR
VACCINES, AND THEN THE
RENAISSANCE OF IMMUNOTHERAPY
WITH CAR-T CELLS, TIL AND
CHECKPOINT BLOCKADE.
CELL THERAPY WAS NOT NEW, IT
GOES BACK TO DONALD THOMAS'
STUDY WITH BONE MARROW
TRANSPLANT, WHICH HE DID IN
COOPERS TOWN, NEW YORK AT THE
MARY IMAGING BASSETT HOSPITAL, A
LITTLE 200-BED COMMUNITY
HOSPITAL.
I'D LIKE TO GIVE YOU A REALLY
QUICK UPDATE ON IL-2 AND HOW IT
CAME ABOUT.
THIS IS A PICTURE OF A WHALE
CRESTING THE GOLDEN GATE BRIDGE.
IT SAT IN THE BOARDROOM FOR
CETUS TRYING TO CLEAN UP OIL
SLICKS TO SAVE THE WHALES.
ALSO CLONED IL-2.
STEVE AND I DID OUR FIRST
STUDIES WITH NATURAL PRODUCT
IL-2 WITH A COLLABORATION WITH
DU PONT WHERE THEY PUT 30 PEOPLE
IN A MA HA TAN-LAKE PROJECT
MAKING IL-2, BUT THAT SAME YEAR,
THEY CLONED IL-2 AND
OUT-LICENSED TO ROCHE.
ROCHE MADE A FAUSTIAN DEAL WITH
CETUS TO TRADE PCR RIGHTS SO
THAT CETUS COULD GET IL-2.
WHEN IT FIRST WENT IN FOR
APPROVAL IN 1990, I REMEMBER THE
MEETING AT THE FDA IN AUGUST,
THEY FAILED TO GET APPROVAL SO
IT WAS ACQUIRED BY CHIRON.
NOVARTIS ACQUIRED CHIRON,
NOVARTIS OUTLICENSED IL-2 TO
PROMETHEUS, PROMETHEUS WAS
ACQUIRED B NESTLE, IT WAS THEN
CLINGEN DID IT, AND MOST
RECENTLY IOVANCE ACQUIRED IT FLO
YE40YEARS LATER IN 2023.
THE GREAT PROMISE OF IL-2 WAS
DRIVEN BY THE FACT THAT YOU
COULD SEE EXTRAORDINARY LONG
LIVED RESPONSES, REALLY PROVING
THAT IMMUNOTHERAPY WORKS, SUCH
AS THIS GENTLEMAN WE TREATED
HERE IN THE BRANCH AT THE
CLINICAL CENTER WHO HAD A LIVER
THAT LOOKED JUST AS BAD AS HIS
LUNGS AND AFTER A SINGLE COURSE
OF IL-2, HAD LONG-LIVED COMPLETE
RESPONSE.
THERE WERE SOME RANDOMIZED
TRIALS ALONG THE WAY.
THE SECOND ONE WAS THIS
RANDOMIZED PHASE 3 TRIAL IN WHAT
WAS CALLED THE LACIL2 WORKING
GROUP THAT WAS SET UP BY VINCE
DAVITA TO TRY AND PROVE THAT
STEVE AND I AND OTHER PEOPLE IN
THE BRANCH WERE NOT CRIMINALS,
THAT WE WERE NOT MAKING UP ALL
THIS STUFF, THAT IT REALLY DID
WORK.
AND THIS WAS A STUDY DONE
SHOWING -- COMPARING HIGH DOSE
IL-2 TO A LOWER DOSE IL-2 AND
INTERFERON REGIMEN.
BUT THAT WAS THE SECOND
RANDOMIZED TREATMENT.
THE FIRST ONE WAS ONE DONE BY
JIM YANG, WHERE HE COMPARED HIGH
DOSE AND LOW DOSE IL-2 AND IL-2Y
CANCER PATIENTS AND SHOWED THAT
THOSE CRs WERE MUCH MORE
DURABLE THAN PATIENTS GETTING
HIGH DOSE IL-2.
SO FOR THOSE -- THIS IS A LITTLE
ADVERTISEMENT, WE JUST DID A
REPORT IN NATURE IMMUNOLOGY ON
CD28.
MY PERCEPTION OF IMMUNOLOGY AND
TUMOR IMMUNOLOGY IS THAT ALL
RHODES LEAD TO CD28 AND COSTIM,
WHETHER THAT BE CHECKPOINTS,
REQUIREMENT FOR CD28 IN CAR-T
CELLS, THE SMALL MOLECULE
INHIBITORS, OR NOW NEW
GENERATION OF COSTIMULATORY
MOLECULES, AND THEY ALL LEAD
ULTIMATELY TO IL-2 PRODUCTION.
SO I LIKE TO SAY THAT THE EARLY
INTUITION THAT IL-2 WAS
IMPORTANT WAS CORRECT.
THIS GOES BACK TO THE SIGNAL
1 SIGNAL 2 OF BRECHER AND COHEN,
THE DISASTROUS USE OF ANTIBODIES
TO CD28 WHICH CAUSED THE
CYTOKINE STORM.
SOME OF YOU MAY BE AWARE OF
THAT.
THE USE OF CHECKPOINT INHIBITORS
WHICH LARGELY WORK BY LIBERATING
CD28 SIGNALING, THE ABILITY OF
CAR-T CELLS TO WORK.
THE FIRST GENERATION DONE BY PAT
HU HERE AT THE BRANCH DIDN'T
WORK ALL THAT WELL BUT WHEN YOU
PUT A CD28 SIGNALING DOMAIN IN,
ALL OF A SUDDEN THEY START
WORKING, WE NOW HAVE A NEW
GENERATION.
IL-2 IS NOT OVER.
JONATHAN WARY IS GOING TO COME
VISIT THE BRANCH, I GUESS, FOR
THE NEXT SITE VISIT, SHOWED THAT
MANY OF THE SENESCENCE MARKERS
SEEN WITH EXPRESSION OF TOX
DRIVING PD-1, LAG3, TIM 3, ET
CETERA CAN BE OVERCOME IN PART
BY PHOSPHOSTAT5 EXPRESSION AS
THE ANECDOTE DRIVEN LARGELY BY
IL-2.
IN A MODEL, IN A MURINE MODEL
WHERE HE ADOPTIVELY TRANSFERRED
CELLS, THOSE CELLS THAT HAD
PHOSPHOSTAT5 CONSTITUTIVELY
ACTIVE WERE MUCH MORE EFFECTIVE
IN MEDIATING ANTITUMOR EFFECTS
THAN THOSE WITHOUT.
AL, TWO GROUPS REPORTED RECENTLY
IN NATURE THAT PG2 INHIBITS IL-2
SIGNALING AND IT DOES THIS
THROUGH A VERY INTERESTING
PATHWAY OF DOWN REGULATING THE
ILIL-2 RECEPTOR GAMMA CHAIN.
SO AGAIN, SIGNALING THE
IMPORTANT ROLE OF IL-2 IN THE
BIOLOGY.
I SHOULD SAY IT'S BEEN A REALLY
LONELY TIME IN SIGHT KINE IN CYY
OSMGHT WE HAVEN'T HAD A NEW
APPROVAL IN CYTOKINE SINCE 1994,
UNTIL THE APPROVAL OF THE IL 15,
ANKTIVA FOR BLADDER CANCER.
I WROTE BOOKS ON CYTOKINES, I
THOUGHT THAT WAS GOING TO BE THE
WAY HOME BUT IT ACTUALLY TURNED
OUT TO BE CELL THERAPY.
MATT KIEVER AND -- CHAMPIONED
THIS IN MOUSE MODELS BUT THE
WORK THAT WAS DONE HERE IN THE
BRANCH WAS SEMINAL.
SOME OF YOU MAY NOT KNOW IT, BUT
THE FIRST CELL THERAPIES WERE
ACTUALLY SENSITIZED BIG
LYMPHOCYTES TO HUMAN TUMORS THAT
NORM WALLMARK DID.
THIS WAS NEVER PUBLISHED.
THEN SUBSEQUENTLY THE LAK
STUDIES THAT WE CONDUCTED
PUBLISHED IN '80, THEN THE THING
THAT OPENED UP THE DOORS, THE
COMBINATION OF IL-2 AND LAK IN
1985 WITH THIS NEW ENGLAND
JOURNAL PAPER.
YUTAKA KAWAKAMI AND I SHOWED
THAT IL 4 WAS INDEED A T-CELL
GROWTH FACTOR, AND IT HELPED
LAUNCH THE TIL PROGRAMS, AND
THEN SINCE THEN, IT'S BEEN
AMAZING THAT THE CELL THERAPY
LANDSCAPE IS REALLY INCREDIBLY
CROWDED AND COMPLEX NOW.
I WAS AT A LITTLE COMPANY CALLED
IOVANCE, I'M GLAD TO SEE SOME OF
MY COLLEAGUES STILL HERE, HELPED
TO DEVELOP THE PROPOSAL THAT
JUST GOT APPROVED THIS LAST
FEBRUARY.
IF YOU LOOK AT ALL THE TIL
COMPANIES, THERE WAS SOMETHING
LIKE 20 OF THEM THAT HAD BEEN
DEVELOPED AGAIN LARGELY COMING
OUT OF WORK BY STEVE AND HIS
COLLEAGUES HERE AT THE BRANCH.
WE GOT OUT OF THE BUSINESS ONLY
BECAUSE OF ITS COSTLINESS.
THERE ARE NEW ENTREES, CONTROL
THERAPEUTICS IN CHICAGO, AND
KENDALL SQUARE, MICHAEL BENSON
IS HERE FROM THERE, BUT I NSTILL
BIOJUST GOT OUT AND ACHILLES
JUST GOT OUT OF THE BUSINESS.
I'D LIKE TO THANK JAMES GULLEY,
WHO HELPED PREPARE ME FOR MY NEW
ROLE, WHICH IS AS NEW EDITOR AND
CHIEF FOR GITSI AND ENCOURAGE
ANY OF YOU WHO HAVE GOOD
MANUSCRIPTS TO SEND THEM MY WAY.
AND THAT'S IT.
THANK YOU FOR YOUR ATTENTION.
[APPLAUSE]
>> WE HAVE A FEW MINUTES SO IF
THERE ARE QUESTIONS, WE CAN TAKE
THEM, OTHERWISE WE'LL GO TO THE
NEXT SPEAKER.
OKAY, SO OUR NEXT SPEAKER IS
GOING TO BE JAMES MULE FROM THE
MOFFITT CANCER INSTITUTE.
>> STEVE, IT'S WONDERFUL TO SEE
YOU AGAIN.
I JOINED THE BRANCH COMING OUT
OF THE FRED HUTCH IN SEATTLE.
I JOINED THE BRANCH IN '83, AND
IT CHANGED MY LIFE
PROFESSIONALLY FOR SURE.
AND SO IT'S A SPECIAL HONOR TO
BE HERE TO CELEBRATE THIS 50TH
ANNIVERSARY.
IT'S REALLY A WONDERFUL
EXPERIENCE.
SO I'M GOING TO GIVE YOU SORT OF
A QUICK HISTORY OF THE
DEVELOPMENT OF IL IL-2
PRECLINICALLY, AND THEN ALSO
TALK ABOUT THE PRE-CLINICAL WORK
WITH LAK CELLS, AND IF TIME
PERMITS, I'LL GIVE YOU A SENSE
OF SOME OF THE THINGS I'M DOING
NOW.
SO WE USED TO CALL THE CYTOKINE
OF THE MONTH CLUB, AND AFTER WE
STARTED THE WORK WITH CETUS, A
NUMBER OF OTHER COMPANIES
STARTED COMING TO DR. ROSENBERG
ABOUT OPPORTUNITIES WITH OTHER
CYTOKINES.
THIS IS JUST A PARTIAL LIST
DURING THE TIME I WAS AT THE
BRANCH.
A NUMBER OF PUBLICATIONS HAD
COME OUT OF THAT.
DEANX, A CORPORATION IN PALO
ALTO WAS INVOLVED, GENETICS
INSTITUTE IN BOSTON AND OTHER
COMPANIES.
BUT I WANT TO -- AS I SAID
EARLIER, I'M GOING TO FOCUS MOST
OF THE TIME ON IL-2.
SOME OF YOU MAY NOT KNOW THIS,
BUT CETUS WAS ACTUALLY THE FIRST
BIOTECH COMPANY ON THE PLANET.
IT WAS NOT GENENTECH ACTUALLY,
CETUS STARTED FIVE YEARS BEFORE
GENENTECH IN THIS SMALL BUILDING
IN BERKELEY, CALIFORNIA.
IT QUICKLY, AS MIKE POINTED OUT,
EXPANDED DURING THE TIME OF
THEIR WORK WITH US WITH THE
ACQUISITION OF CETUS BY KI CHIR.
THIS WAS THE BUILDING IN
EMERYVILLE WHICH MANY OF US HAD
SPENT TIME VISITING.
NOW YOU LOOK TODAY AT EMERYVILLE
AND YOU CAN SEE THE CIRCLE WHERE
THE OLD CETUS CHIRON BUILDING
USED TO BE, AND THE REMARKABLE
BIOTECH EXPLOSION THAT HAS OCC
OCCURRED OVER THE YEARS.
IT'S NOW CALLED EMERY YARDS.
SO THIS LAUNCHED EVERYTHING,
THIS PAPER WITH STEVE IN SCIENCE
WITH COLLEAGUES AT CETUS.
WHICH WAS THE FIRST USE OF
RECOMBINANT IL-2 PRODUCED IN
E. COLI.
WE WERE OFF TO THE RACES.
THIS WAS ONE OF THE FIGURES IN
THE PAPER, THE CLOAPING OF THE E
IL-2 GENE BY ACRYLAMIDE GEL, IT
WAS ABLE TO BE PURIFIED IN THE
COMPANY USING THESE MEGA COLUMNS
THAT STOOD FROM THE FLOOR TO THE
CEILING OF THREE STORIES IN THE
BUILDING.
BUT STEVE MAY REMEMBER THIS.
THE FIRST OR SECOND OR THIRD LOT
THEY SENT US, WE PUT IN MICE AND
ALL THE MICE DIED.
AND IT WAS BECAUSE OF THE
CONTAMINATION WITH ENDO TOXIN,
WHICH, AGAIN, THEY QUICKLY TOOK
CARE OF THAT PROBLEM AND SENT US
THE BEST STUFF.
AND YOU CAN SEE THE 15KD CLEANED
UP SAMPLE OF IL-2.
SO STEVE SHOWED THAT THE
MATERIAL COULD, IN FACT,
REPLICATE WHICH WAS DONE IN
NATURAL IL-2, THE CREATION OR
GENERATION OF LAK CELLS, BOTH IN
THE MOUSE AND THE HUMAN.
AND AS I MENTIONED, THIS WAS A
REVOLUTION.
I MEAN, TO LOOK AT THE ABILITY
OF THIS EQUIPMENT IN THE COMPANY
IN THE EARLY YEARS OF BIOTECH
WAS PRETTY ASTONISHING.
THIS WAS ONE EXAMPLE OF THE
PRODUCTION OF THE E. COLI AT THE
COMPANY.
STEVE MAY NOT HAVE SEEN THIS
ONE, BUT THIS WAS -- I TOOK A
LIST OF ALL THE IL-2 THAT WAS ON
THE FREEZER DOOR OUTSIDE THE
LAB, AND THE LOT NUMBERS ARE
SHOWN, DURING THE TIME I WAS IN
THE BRANCH.
NOW THINK ABOUT THIS.
FOUR TIMES 10 TO THE 11TH
INTERNATIONAL UNITS OF IL-2 WERE
USED DURING THAT TIME.
AND THIS WAS PURELY FOR THE
PRE-CLINICAL WORK.
AND I REMEMBER STEVE SAYING TO
ME ONE DAY, HE SAYS, JIM, IF WE
EVER COME BACK AS MICE, WE'RE IN
TROUBLE.
[LAUGHTER]
SO THEN WE WENT ON WITH THIS
JOURNAL OF EXPERIMENTAL MEDICINE
PAPER, WHICH REALLY SHOWED THE
ABILITY OF RECOMBINANT IL-2 AT
HIGH DOSE TO IMPACT ESTABLISHED
TUMORS IN MICE.
AND THIS WAS THE RESULT OF 36
CONSECUTIVE EXPERIMENTS THAT
WE'VE SHOWN IN THE PAPER WITH
THREE-DAY EARLY MULL KNOWARY
METASTASES.
YOU CAN SEE THAT MORE IL-2,
BETTER EVENT IN MICE.
-- BETTER EFFECT IN MICE.
AND THAT'S WHERE I THINK WE CAME
UP WITH THE ONE MORE DOSE LOTZE
FOR IL-2.
AND THIS WAS 15 CONSECUTIVE
EXPERIMENTS IN ALREADY ESTABL
ESTABLISHED LUNG TUMORS WITH
SARCOMA, AND AGAIN, YOU CAN SEE
THE EFFICACY OF RECOMBINANT IL-2
IMPACTING AND CAUSING REGRESSION
OF ACTUALLY ESTABLISHED CANCERS.
AND THIS WAS SHOWN AS WELL NOT
ONLY IN THE LUNGS BUT
SUBCUTANEOUS TUMORS.
YOU COULD SEE THE REGRESSION OF
ALREADY ESTABLISHED TUMORS AT
SUBCUTANEOUS SITES.
WITH STEVE EDINGHAUSEN JOINING
US IN THE LAB, STEVE HAD HIM ON
THIS PROJECT, WAS TO LOOK AT
STIMULATION OF LYMPHOCYTES
WITHIN THE MOUSE WHEN ONE WAS
GIVEN IL-2 TO START TO GET A
SENSE OF THE MECHANISMS INVOLVED
IN THE REGRESSION OF TUMORS BY
THAT INTERLEUKIN.
I WON'T GO THROUGH ALL THE
SPECIFICS OTHER THAN TO SAY THAT
HE DID SHOW THAT IL-2 COULD
CAUSE THE PROLIFERATION OF
LYMPHOCYTES IN THE
IMMUNE-RELATED ORGANS, BUT AS
YOU WOULD EXPECT, NOT ELSEWHERE
AND SO FORTH.
SO IL-2 COULD CAUSE SIGNIFICANT
PROLIFERATION IN THE ANIMAL
SYSTEMICALLY AT MULTIPLE
LYMPHOID ORGAN SITES.
HE ALSO SHOWED NOT ONLY DID IT
CAUSE PROLIFERATION BUT IT
PRODUCED LAK CELLS IN VIVO.
SO THIS WAS TO SHOW THAT YOU
COULD TAKE CELLS OUT OF THOSE
MICE, EXPOSE THEM TO FRESH TUMOR
CELLS IN CULTURE AND SHOW THAT
THEY COULD, IN FACT, LYSE
MULTIPLE TYPES OF FRESH TUMOR
CELLS.
WE THEN WENT ON TO GIVE A MORE
IN DEPTH ANALYSIS OF WHAT WAS
GOING ON IN THERAPEUTIC EFFICACY
OF MICE.
THIS TO ME WAS ONE OF THE MORE
ASTONISHING FINDINGS THAT WE HAD
WHERE STEVE ASKED US TO COMPARE
NON-IMMUNOGENIC SARCOMAS THAT WE
HAD IN THE LAB, WHICH WAS MCA101
AND 102, WHICH WAS
NON-IMMUNOGENIC DEFINED BY THE
INABILITY OF THESE TUMOR CELLS
BY DIFFERENT MEANS TO IMMUNIZE
THE MICE TO REJEBT VIABLE TUMOR.
SO THESE WERE CLASSICALLY
NON-IMMUNOGENIC.
AND MCA-105 AND 106, WHICH WERE
CLASSICALLY DEFINED AS WEAKLEY
IMMUNOGENIC.
GIVE IL-2 AND YOU COULD SEE THAT
THREE DAY METS, RARTLESS IF THE
TEU NOR WAS NON-IMMUNOGENIC OR
IMMUNOGENIC, YOU COULD CAUSE
REGRESSION.
HOWEVER, WITH THE MORE
ESTABLISHED TUMORS, IT WAS ONLY
THE WEAKLY IMMUNOGENIC TUMORS
THAT RESPONDED TO HIGH DOSE IL-2
AND DURING THAT 10-DAY PERIOD,
ALLOWED THE IMMUNE SYSTEM TO
GENERATE T CELLS THAT THEN COULD
REACT TO THE IL-2 AND CAUSE
REGRESSION.
SO IN THIS STUDY, IMMUNOGENIC
TUMORS, IL-2 WAS EFFECTIVE.
NON-IMMUNOGENIC TUMORS, ONLY
EARLY ON DURING THE EARLY
METASTATIC STAGE, AND THOSE WERE
DUE TO THE PRO TU PRODUCTION OFK
CELLS.
SO LAK CELLS EARLY ON, WE SHOWED
YOU COULD ELIMINATE THESE LAK
CELLS WITH ANTI-ACGM1, AND YOU
COULD LOSE THE EFFECT ON
THREE-DAY METS OF HIGH DOSE
IL-2.
THE OPPOSITE WAS TRUE ON 10-DAY
METS SO IF YOU ELIMINATE IT, AT
THAT TIME IT WAS CALLED LY2, WAS
THE MARKER FOR CD8 CELLS.
SO WE HAD THE ABILITY TO
ELIMINATE THOSE CELLS IN MICE,
AND THAT ELIMINATED THE IMPACT
OF HIGH DOSE IL-2 ON ESTABLISHED
TUMORS.
SO THAT REALLY MOCKED DOWN THE
ABILITY OF US TO DISTINGUISH THE
IMPACT OF LAK CELLS EARLY ON
WITH T CELLS LATER ON DURING THE
PROGRESSION OF TUMOR.
ALONG THE SAME PATH, STEVE ASKED
ME TO WORK WITH HIM AND OTHERS
ON LAK CELLS AND THE ABILITY TO
USE THOSE AT LOWER DOSES OF IL-2
AND TO SEE IF THAT COULD IMPACT
ON TUMORS IN MICE.
SO THIS WAS A PAPER IN SCIENCE
IN 1984.
AND, IN FACT, YOU COULD SEE HERE
THE LAK CELLS BY THEMSELVES HAD
SOME IMPACT ON EARLY TUMOR, BUT
WITH IL-2, YOU COULD SEE A
DRAMATIC REDUCTION IN THE NUMBER
OF METASTASES.
AND THIS WAS THE FIRST PHOTO
THAT WAS SHOWN OF THE EFFICACY
OF LAK CELLS IN LUNG TUMORS.
IN
THAT SCIENCE PAPER.
THIS WENT ON TO SHOW THAT YOU
COULD GENRES LAK CELLS AS EARLY
AS -- IN CULTURE, SO NOW I'M
GOING TO END JUST BY SHOWING YOU
A SNAPSHOT OF WHAT WE'VE BEEN
DEVELOPING SINCE.
SO I'VE BEEN INTERESTED IN
CHEMOKINES FOR A NUMBER OF
YEARS, AND I WAS ASKED BY JCO TO
HAVE A COMMENTARY ON A PAPER BY
WOLF FRIDMAN'S GROUP WHICH
SHOWED THE ACTIVITY OF WHAT WAS
CALLED ECTOPIC LYMPH NODES AT
THE TIME, CONNECTING TO CLINICAL
RESPONSE IN LUNG CANCER.
SO WHEN I WAS ASKED TO HAVE THE
COMMENTARY, I DID A QUICK
SEARCH, AND IF YOU GOOGLE THAT
WITH WHATEVER SEARCH ENGINE YOU
USE AND YOU PUT IN TERTIARY
LYMPHOID STRUCTURES OR ECTOPIC
LYMPH NODE TUMORS, NOTHING CAME
UP.
FAST FORWARD TODAY, YOU CAN SEE
THE EXPLOSION OF ACTIVITY OF
INDIVIDUALS NOW WORKING TO
UNCOVER WHAT IT MEANS TO HAVE
THESE TERTIARY LYMPHOID
STRUCTURES IN SOLID TUMOR
MASSES.
SO WITH MERCK BIOINFORMATACIST,
WE HIT ON A GENE EXPRESSION
SIGNATURE YEARS AGO.
THIS IS 12 YEARS AGO WHEN WE
PUBLISHED THIS, THAT COULD
DISTINGUISH TUMORS AS YOU CAN
SEE GREEN VERSUS RED.
THIS WAS FROM THE META GENE
GROUPING OF ABOUT 96 GENES THAT
MERCK UNCOVERED.
WITHIN THAT GROUPING WERE A HOT
SPOT OF 12 CHEMOKINES.
WE THEN TOOK IT ACROSS THOUSANDS
OF SOLID TUMOR ARRAYS.
THIS SHOWS ONE EXAMPLE IN
MELANOMA, WHICH, IN FACT, THE
SIGNATURE COULD RELIABLY
IDENTIFY WITHIN MELANOMA
COLORECTAL CANCER, FRIDMAN USED
IT, TERTIARY STRUCTURES WHICH
YOU SEE SURROUNDING A TUMOR MASS
IN A HUMAN MELANOMA.
YOU KNOW, WHEN WE FIRST
UNCOVERED THIS, I ASKED THE
PATHOLOGIST, I SAID HAVE YOU
EVER LOOKED AT THIS, HAVE YOU
EVER SEEN THIS?
THEY SAID NO.
I SAID, WELL, WHY NOT?
THEY SAID, WELL, NO ONE EVER
ASKED US TO LOOK FOR THEM.
AND HISTORICALLY THEY WOULD
SCORE IMMUNE ILL FILTRATE FROM A
1 PLUS TO A 4 PLUS.
SO NOW YOU CAN SEE THAT THESE
UNIQUE STRUCTURES ACTUALLY MIMIC
PERIPHERAL LYMPH NODES TO
SOME DEGREE WHERE YOU SEE A
DENSE B CELL FOLLICLE IDENTIFIED
BY CD20 SURROUNDED BY T CELLS,
MARGINS AROUND T CELLS, 4s AND
8s, AS WELL AS THE UNDERLYING
FOLLICULAR RETICULAR SITES OR
DENDRITIC CELLS.
THIS PAPER WILL BE APPEARING IN
NATURE MEDICINE.
WE CONDUCTED A MOFFITT -- A
CLINICAL TRIAL WITH AN ONCOLYTIC
VIRUS WITH A COMPANY CALLED CG
ONCOLOGY WITH PEMBRO.
DRAMATIC REGRESSION OR
ELIMINATION OF A
NON-MUSCLE-INVASIVE BLADDER
CANCER BY THIS THERAPY, WHICH IN
THIS PAPER SHOWS A DIRECT
LINKAGE WITH THE ACTIVITY AND
PRESENCE OF TLSs IN THOSE
BIOPSIES.
SO I'LL END HERE BY SAYING
CONGRATULATIONS, DR. ROSENBERG,
AND IT A DELIGHT TO BE HERE.
SO THANK YOU VERY MUCH.
[APPLAUSE]
>> WE'RE DOING GREAT ON TIME.
IF ANYBODY HAS A QUESTION, WE
COULD TAKE ONE NOW.
ALL RIGHT.
SO LET'S MOVE ON TO OUR NEXT
SPEAKER THEN.
WE HAVE STEPHANIE GOFF, A MEMBER
OF OUR CURRENT SENIOR STAFF HERE
IN THE SURGERY BRANCH, TO TALK
ABOUT THE EVOLVING THERAPEUTIC
LANDSCAPE OF MELANOMA.
>> THANK YOU ALL.
I HAVE BEEN TASKED WITH SORT OF
GIVING YOU AN OVERVIEW OF ALL OF
THE FANTASTIC WORK, STARTED
OBVIOUSLY BY DR. ROSENBERG IN
CONJUNCTION WITH MIKE AND JAMES
AND HOW IT REALLY HAS CHANGED
THE LANDSCAPE FOR MELANOMA OVER
THE COURSE OF THE 50 YEARS THAT
HE'S BEEN HERE.
SO THE FIRST THING IS THAT IT'S
GREAT NEWS, RIGHT?
MORTALITY FOR PATIENTS WITH
METATATIC MELANOMA HAS BEEN
DROPPING.
UNFORTUNATELY, THAT WASN'T DUE
TO INTERLEUKIN NECESSARILY
BECAUSE OF THE SMALL RATES, BUT
REALLY THE REVOLUTION THAT
HAPPENED WHEN CHECKPOINT
INHIBITORS CAME ON BOARD.
I'M NOT GOING TO EMBARRASS
MYSELF ABOUT TRYING TO TALK TO
YOU ABOUT THAT WHEN WE HAVE
OTHERS TO TALK ABOUT THAT
TOMORROW.
BUT I DID WANT TO TALK YOU
THROUGH SORT OF HOW LONG THIS
TAKES TO HAPPEN.
WHEN DR. ROSENBERG FIRST GOT
HERE, DECASH SEEN WAS REALLY THE
ONLY THING BEING USED FOR
PATIENTS WITH METATATIC
MELANOMA.
THE ONE-YEAR SURVIVAL RATE WAS
IN THE 5 TO 10%.
IT TOOK FROM 1985 WHEN THAT
FIRST CLINICAL EXPERIENCE WAS
PUBLISHED TO 1998 TO GET
INTERLEUKIN 2 APPROVED FOR
PATIENTS WITH METATATIC
MELANOMA.
CTLA4 WAS FIRST DESCRIBED OF AS
1987.
THE FIRST CLINICAL TRIAL STARTED
HERE IN 2001, WASN'T PUBLISHED
UNTIL 2003 AND TOOK AN
ADDITIONAL EIGHT YEARS TO GET
THAT APPROVED BY THE FDA.
PD-1 WAS FIRST DESCRIBED IN
1992, FIRST CLINICAL EXPERIENCE
WASN'T PUBLISHED FOR 10 YEARS
AND IT TOOK FORTUNATELY JUST A
FEW SHORT YEARS FOR THAT TO GET
APPROVED IN BOTH ITS VERSIONS.
WE LEARNED A LOT ABOUT TUMORS
DURING THIS TIME.
NO ONE WOULD HAVE EVER THOUGHT
ABOUT SEQUENCING A TUMOR BACK IN
1975 BUT IN 2002 WE STARTED TO
UNDERSTAND ABOUT ACTIVATING BRAF
IN PATIENTS WITH MELANOMA AND
THE FIRST CLINICAL EXPERIENCE
WITH THE BRAF DRUGS WAS IN 2010.
IT WAS ABLE TO GET APPROVAL IN
2011.
THEN THE COMBINATIONS OF THOSE
THERAPIES STARTED TO GET
APPROVED.
2016, 2020, 2022, THE ADDITION
OF LAG 3.
THROUGH ALL OF THIS DEVELOPMENT
AS ALL OF THIS WAS HAPPENING IN
THE FIELD OF MEL N MELANOMA, THE
BRANCH TO BEGIN WITH AND THEN
OTHERS, MOSTLY PEOPLE WHO HAD
BEEN AT THE BRANCH AND
METASTASIZED ACROSS THE WORLD,
THEN LED TO THIS GREAT
ACHIEVEMENT THIS PAST YEAR WHEN
IOVANCE WAS ABLE TO GET THE
FIRST TIL THERAPY APPROVED FOR
PATIENTS WITH METATATIC
MELANOMA.
SO WHY?
YOU KNOW, YOU'VE HEARD US TALK
ABOUT LAK CELLS, YOU'VE HEARD
THE WORK ABOUT IL-2 AND
CYTOKINES.
A BEAUTIFUL THING ABOUT MELANOMA
IS WHEN YOU LOOK AT THE TUMORS,
ALMOST ALL OF THEM ARE HEAVILY
IMMUNE ILL FILTR INFILTRATEED WH
CD3, ALL THESE BEAUTIFUL T CELLS
LIVING IN THERE.
SO THE SCHEME THAT MANY OF US
HAVE COME TO KNOW SO WELL, THE
CIRCLE OF LIFE.
YOU EXCISE A TUMOR, YOU PLATE
THE FRAGMENTS, YOU THEN TRY TO
CULTURE THOSE IN HIGH DOSE IL-2,
YOU TEST FOR SOME REACTIVITY.
SO WE DON'T ALWAYS PLATE THEM.
ANYBODY WHO'S BEEN HERE OVER THE
TIME KNOWS THAT WE'VE
EXPERIMENTED WITH DIGEST THAT WE
THEN PLATED, WE EXPERIMENTED
WITH BAGS THAT WE GREW THEM IN,
WE EXPERIMENTED WITH THE GAS
PERMEABLE FLASKS.
OUR CURRENT ME THIS HA METHOD IK
TO THE FRAGMENTS.
IN THE DAYS OF MELANOMA, WE
WOULD TEST AGAINST CELL LINES
THAT HAD BEEN VERY THOROUGHLY
CHARACTERIZE.
CHARACTERIZED.
WE COULD EXPRESS WHICH ONES WE
WANTED.
NOW IN THOSE DAYS WE THOUGHT WE
WANTED THE MELANOMA
DIFFERENTIATION ANTIGEN SO WE
LOOKED FOR REACTIVITY AGAINST
THINGS LIKE MART, WE WOULD ALSO
BE ABLE TO GROW THE PATIENT'S
OWN TUMOR CELL LINE, A GREAT
MODEL TO TEST TO SEE IF THE TIL
WERE REACTIVE TO THAT.
WE WOULD PICK THE FRAGMENTS WE
WANTED, EXPAND THEM UP TO THE
ORDER OF 10 TO THE TENTH, AND
THEN GIVE THE PATIENT A LITTLE
BIT OF CHEMOTHERAPY IN THE
BEGINNING, A LOT OF CHEMOTHERAPY
LATER ON, AND THEN GIVE THE
PATIENTS THEIR CELLS AND IL-2.
SO THE FIRST PUBLICATION OF
THIS, 1988, THESE WERE PATIENTS
WITH METASTATIC MELANOMA, AND
YOU CAN SEE THE BEFORE AND AFTER
CHEST X-RAYS HERE.
THIS IS PRE-C T SCAN.
SO IF YOU CAN IMAGINE DOING ALL
THE ADVANCED CLINICAL WORK WE DO
TODAY, TRYING TO MAKE
MEASUREMENTS OFF CHEST X-RAYS
AND CHEST TOMOGRAMS.
9 OF THE 15 PATIENTS NAIVE TO
IL-2 HAD RESPONSE TO THIS
THERAPY, AND TWO OF THE FIVE
PATIENTS HAD BEEN REFRACTORY TO
IL-2, ALSO DEVELOPED A RESPONSE
TO THIS THERAPY.
BUT UNFORTUNATELY, THOSE
RESPONSES WERE NOT VERY DURABLE.
SO THE NEXT QUESTION WAS, A
SINGLE DOSE OF CYCLOPHOSPHAMIDE
DIDN'T GIVE US SORT OF THE
IMMUNE MILIEU THAT WE WANTED.
INCREASE THE LYMPHODEPLETION, AT
FLEW DARE BEAN, GET RID OF THE
ENDOGENOUS T CELLS AND THEN GIVE
THESE CELLS AGAIN.
THIS WAS PUBLISHED BY DR. DUDLEY
IN 2005.
YOU CAN SEE A PATIENT WITH
DISEASE IN THE LUNG AND IN THE
LEG THAT DISAPPEARED.
49% OBJECTIVE RESPONSE RATE, AND
MORE IMPORTANTLY, A 12% COMPLETE
RESPONSE RATE.
SO WHEN YOU COMPARE THAT, IL-2
WAS RUNNING IN A COMPLETE
RESPONSE RATE OF ABOUT 4%.
LYMPHODEPLETION WORKED.
IT HELPED MAKE THESE THINGS
DURABLE.
IS MORE LYMPHODEPLETION BETTER?
SO A NUMBER OF SEQUENTIAL
STUDIES WERE DONE.
MANY OF YOU TOOK PART IN A LOT
OF THESE STUDIES.
AND IN 2011, DR. ROSENBERG
PULLED TOGETHER THE 93 PATIENTS
THAT WE TALKED ABOUT FOR SO MANY
YEARS.
43 OF THEM HAD GOTTEN STANDARD
NON-MYELOABLATIVE CHEMOTHERAPY.
ANOTHER GROUP HAD GOTTEN 200
SENT DWRAID OF TOTAL BODY EE
RADIATION TO DEEPEN THEIR
LYMPHODEPLETION AND ANOTHER
GROUP HAD GOT A HIGHER DOSE THAT
REQUIRED STEM CELL RESCUE.
SO WHAT WE YOU A SAW WAS THAT
THE OVERALL RESPONSE RATE, THE
PARTIAL RESPONSE RATE WAS NOT
THAT DIFFERENT BUT THE COMPLETE
RESPONSE RATE APPEARED TO JUMP.
IT SEEMED LIKE YOU DID NEED THAT
DEEPER LYMPHODEPLETION EVEN
THOUGH IT DID COME AT THE COST
OF SOME EXTRA SIDE EFFECTS.
THESE AGAIN WERE SEQUENTIAL, AND
IN THE MATTER OF ALL GOOD
SCIENCE, YOU HAVE TO GO BACK AND
THEN TEST IN A RANDOMIZED
FASHION.
WHEN WE DID THAT, IT TURNED OUT
YOU DON'T ACTUALLY NEED THE
LYMPHODEPLETION, WHICH TURNED
OUT TO BE A GREAT THING FOR OUR
PATIENTS.
YOU DON'T NEED THAT TBI.
THE RESPONSE RATES WERE
IDENTICAL, THE COMPLETE RESPONSE
RATES WERE NEARLY IDENTICAL.
AND WE PUBLISHED THIS IN 2016.
WE ALL KNOW WHAT THESE RESPONSES
CAN LOOK LIKE.
IT CAN HAPPEN IN THE
SUBCUTANEOUS DISEASE, IT CAN
HAPPEN IN VISCERAL DISEASE IN
THE LIVER, AND IT CAN EVEN
HAPPEN IN THE BRAIN.
THIS IS A YOUNG MAN JUST 12 DAYS
LATER, YOU SEE THAT LARGE MET IS
GONE.
HE HAD UPWARDS OF 40 OF THEM ALL
OVER HIS BODY, ALL OF WHICH
RESOLVED.
HE ASKED US TO STOP CALLING HIM
ABOUT SEVEN YEARS LATER, SO I
HOPE HE'S STILL OUT THERE ALIVE
WITHOUT DISEASE.
BUT HE NO LONGER WANTED TO BE A
PATIENT.
NOT THAT I CAN BLAME HIM.
THIS GENTLEMAN WITH LIVER
DISEASE, YOU CAN SEE SEVEN YEARS
LATER, THE LIVER IS CLEAR.
AND THIS GENTLEMAN WITH BRAIN
METS, FOUR OF THEM, THAT YOU CAN
SEE ENHANCED BY CONTRAST, ALL
RESOLVED WITHIN THREE MONTHS.
THESE REGRESSIONS WERE DURABLE,
AND THAT'S REALLY BEEN THE
HALLMARK OF THIS.
SO WHEN WE LOOKED AT ALL OF OUR
PATIENTS HERE AT THE NCI THAT
GOT CELLS GROWN IN A SIMILAR WAY
BY OTHER FRAGMENTS OR DIGESTS
THAT WERE GROWN IN PLATES, WE
LOOKED AT 226 OF THEM AND WE HAD
A 22-MONTH SURVIVAL WITH A
5-YEAR SURVIVAL OF 35%.
BUT WHAT WAS MOST IMPRESSIVE, AT
LEAST TO ME, IS THAT GREEN LINE.
OUR COMPLETE RESPONDERS HAD A
10-YEAR SURVIVAL RATE OF 96%.
SO WE WENT INTO THAT COHORT TO
TRY TO DECIDE WHAT MADE PATIENTS
GET TO THAT GREEN LINE.
WELL INFUSION PRODUCTS THAT HAD
MORE CELLS APPEARED MORE LIKELY
TO BE ABLE TO MEDIATE RESPONSE.
WE IMREU AS MANY AS WE COULD.
MAYBE IT WAS AN UNDERLYING
FEATURE THAT WE WERE ABLE TO
GROW SO MANY CELLS BUT THERE WAS
A SIGNIFICANT DIFFERENCE BETWEEN
OBJECTIVE RESPONDERS AND
NON-RESPONDER WHEN IS YOU LOOK
BY MEDIAN AND IF YOU START TO --
IT OUT IN GROUPS BY 2 TO THE 10,
WHERE THE HIGHER OBJECTIVE
RESPONSE RATE IN THE HIGHEST
GROUP OF CELLS.
WHAT WE DID FIND WAS THAT PRIOR
THERAPY HAD A BIG IMPACT ON
WHETHER OR NOT THESE PATIENT
WOULD DEVELOP A RESPONSE TO TIL.
IF YOU LACKED AT OUR PATIENTS LT
RECEIVED IPIMIBULAB AND COMPARED
THEM TO THOSE THAT WERE
REFRACTORY, THERE WASN'T A
DIFFERENCE IN RESPONSE RATE,
ABOUT 49 TO 60%.
HOWEVER IF YOU LOOKED AT
PATIENTS WHO RECEIVED ANTIPD #
AND YOU LOOKED AT THE GROUP THAT
WAS NAIVE YOU HAD A 56 RESPONSE
RATE WITH A HEFTY NUMBER OF
COMPLETE RESPONDERS.
WHEN YOU LOOKED AT THE SMALLER
GROUP OF 34 PATIENTS THAT HAD
GOTTEN ANTIPD-1 BEFORE YOU HAD A
MUCH LOWER COMPLETE RESPONSE
RATE AND AT THAT TIME ONLY ONE
COMPLETE RESPONSE.
SIMILARLY WITH THE PATIENTS WHO
HAD BRAF MUTATED TUMORS, IF YOU
LOOKED AT THOSE THAT HAD
RECEIVED MAP KINASE INHIBITORS,
YOU HAD A LOWER RESPONSE RATE IN
THOSE PATIENTS THAT WERE
REFRACTORY.
AND THAT IMPACTED ON THEIR
SURVIVAL AS WELL.
YOU CAN SEE THE DIFFERENCES
BETWEEN THE PATIENTS THAT WERE
NAIVE AND REFRACTORY IN THESE
CURVES.
NOW THE PRESENCE OR ABSENCE OF
THAT ACTIVATING BRAF MUTATION
DIDN'T HAVE AN EFFECT ON WHETHER
TIL THERAPY WAS ABLE TO BE
USEFUL, BUT IF THEY HAD GOTTEN
THE DRUGS TO TRY TO BREAK DOWN
THE BRAF MEK THEN YOU SAW THE
DIFFERENCE IN RESPONSE.
HOWEVER, COMPLETE RESPONSES WERE
STILL POSSIBLE WHEN YOU USED TIL
AFTER PD-1.
THIS IS A GENTLEMAN WHO HAD GONE
THROUGH COMBINATION THERAPY, HAD
DEVELOPED A GROWING PARARENAL
MASS THAT WAS RESENTED AS THE
SOURCE OF HIS TIL.
AT THE TIME HE WAS TREATED YOU
SEE THE LUNG DISEASE THERE AS
WELL AS THE DISEASE THAT WAS
SORT OF WRAPPING AROUND THE
ILIAC BONE AND YOU CAN SEE FIVE
YEARS LATER, THAT DISEASE IS
GONE, AND IN FACT THE BONE HAS
RECALCIFIED.
SIMILARLY, THIS IS A GENTLEMAN
WHO CAME TO US WITH UNMANAGEABLE
AXILLARY DISEASE TO THE POINT HE
COULDN'T LOWER HIS ARMS, IT WAS
AFFECTING HIS DAILY ACTIVITY,
AND CAN YOU SEE WE RESPECTED A
SMALL NODULE OF THAT TUMOR
SEPARATE FROM THE REST,
DEVELOPED TIL FROM THAT, AND
EIGHT YEARS LATER, HE HAS A
CLEAN PET SCAN.
AND ACTUALLY HAS JUST BEEN
RECENTLY DISCHARGED FROM THERAPY
BECAUSE HE, TOO, NO LONGER WANTS
TO BE A PATIENT.
SO TIL FOR CHECKPOINT REFRACTORY
METATATIC MELANOMA IS STILL
EFFECTIVE.
THIS IS THE RESULTS THAT CAME
OUT OF THE IOVANCE STUDY OF
66 PATIENTS DEMONSTRATING A 22
MONTH SURVIVAL AND A 5-YEAR
SURVIVAL THERE.
SIMILARLY ACROSS THE POND IN
EUROPE, THERE WAS A RANDOMIZED
TRIAL OF 'TIL VERSUS IPIMIBULAB
IN PATIENTS WITH METATATIC
MELANOMA.
WE HAD A 25-MONTH SURVIVAL AN
OBJECTIVE RESPONSE OF 49%.
THESE ARE NOW APPROVED BOTH HERE
IN THE UNITED STATES OBVIOUSLY
AS WE ALL KNOW IN FEBRUARY, AND
FOR USE THROUGH THE EUROPEAN
HEALTH AGENCY IN I BELIEVE
DENMARK AND COPENHAGEN.
SO WHAT IS THE IMPACT OF THAT
PRIOR THERAPY?
ONE OF OUR FELLOWS WENT BACK TO
ALL THE INDIVIDUAL FRAGMENTS
GROWN FOR ALL OF THOSE
INDIVIDUAL PATIENTS, SO WHAT
YOU'RE LOOKING AT HERE ARE
MEDIANS OF MEDIANS.
SO PATIENT MAY HAVE HAD ANYWHERE
FROM 2 TO 24 FRAGMENTS GROWN FOR
THEM AND IT LOOKED AT THE
PERCENTAGE -- HE LOOKED AT THE
PERCENTAGE OF CD3s AS WELL AS
CD8 AND NK CELLS.
IT WAS AFFECTED BY PD-1.
THERE WERE FEWER CD3s, FEWER
CD8s AND MORE NKs IN THOSE.
INTERESTINGLY, ANTICTLA4 DIDN'T
HAVE THAT SAME EFFECT.
THERE WERE NO DIFFERENCES IN THE
BULK PHENOTYPE OF THE INITIAL
TIL PRODUCTS THAT WE WERE
GROWING.
BUT THE INITIAL TIL PRODUCTS ARE
NOT ALWAYS THE FINAL TIL
PRODUCT.
SO WHEN YOU LK AT THE FINAL TIL
PRODUCT, WHAT WE FOUND WAS THAT
PATIENTS THAT WERE PD-1
REFRACTORY HAD MORE CD4 CELLS IN
THEIR BAGS, AND THAT WAS THE
ONLY REAL DIFFERENCE BETWEEN THE
TWO GROUPS.
TIL GREW EQUALLY WELL UNDER THE
UNIFORM CONDITIONS WITH MORE CD.
WHEN YOU LOOK AT THE GENOMICS OF
THE TUMOR, THIS WAS DONE BY
ANOTHER FELLOW HERE IN THE
AUDIENCE TODAY, THE TMB OF THESE
TUMORS IS LOWER.
EVERYONE SHOULD HAVE EXPECTED
THAT.
WE'RE ONLY GETTING PATIENTS NOW
THAT ARE REFRACTORY TO PD-1.
WHO ARE THE PATIENTS THAT ARE
RESPONDING TO PD-1?
THOSE THAT HAVE HIGH TMB.
SO WE'RE ONLY GETTING THE
PATIENTS FOR WHOM THAT THERAPY
HAS NOT WORKED.
SO BY OUR REFERRAL PATTERN, OUR
TMB IS LOWER.
WHAT WE'VE ALSO NOTICED, THOUGH,
IS THAT IT AFFECTS THE ABILITY
TO FIND NEOANTIGENS.
I'M NOT GOING TO GO INTO THE
DETAILS OF HOW WE FIND OUR
NEOANTIGENS.
YOU'RE GOING TO HEAR FROM A LOT
OF DIFFERENT SPEAKERS LATER HOW
WE'VE TAKEN THIS FROM MELANOMA
INTO OUR EPITHELIAL CANCER
SPACE.
INTERESTINGLY, ANNA PASETTA HAD
GONE BACK TO THE INFUSION BAGS
OF A NUMBER OF PATIENTS.
SHE ACTUALLY LOOKED AT THE MOST
FREQUENT CLONES IN THOSE BAGS,
RECONSTRUCTED THE TCRs THAT
CAME FROM THEM AND THEN TESTED
THEM TO SEE WHETHER THEY WERE
TISSUE CULTURE REACTIVE, SHARED
ANTIGEN REACTIVE, MUTATION
REACTIVE.
THE ONLY ONE OF THESE PATIENTS
THAT WAS PD-1 REFRACTORY WAS THE
PATIENT FOR WHOM SHE COULD NOT
IDENTIFY ANY REACTIVITY.
SO THAT'S JUST AN ANECDOTE BUT
MAYBE A HINT OF WHAT WE MAY BE
FACING IN THESE PATIENTS THAT
ARE ANTIPD-1 REFRACTORY.
SO WHEN IT COMES TO THE WHOLE
EVOLVING LANDSCAPE OF OUR
PATIENTS WITH METASTATIC
MELANOMA, THESE WERE THE
THERAPIES AVAILABLE TO THEM NOW.
ANTIPD-1, DUAL CHECKPOINT
BLOCKADE, TRIPLET THERAPY FOR
THOSE PATIENTS THAT HAVE BRAF
MUTATIONS, OR TIL.
CAN YOU SEEYOU CAN SEETHE RYE SL
VERY SIMILAR, YOU CAN SEE THE
COMPLETE RESPONSE RATES THERE,
ALL VERY SIMILAR.
YOU SEE THE GRADE 3 ADVERSE
EVENTS.
THAT HE IS WHERE TIL WILL ALWAYS
TAKE A HIT.
WE DESIGN OUR THERAPY -- ALL OF
OUR PATIENTS WILL HAVE A GRADE
III/4 AE.
WHEN YOU STOP TO LOOK AT THOSE
ONLY DUAL TO THE CELL PRODUCT
AND PERHAPS THE IL-2, YOU'RE
LOOKING AT A MORE REASONABLE
36%.
AND AGAIN THE HALLMARK.
THE DURABILITY OF THIS IS THE
FIVE-YEAR OVERALL SURVIVAL.
SO WHEN YOU LOOK AT THE
QUALITATIVE THINGS THAT OUR
PATIENTS ARE THINKING ABOUT, GO
TO AN INFUSION CENTER EVERY TWO
TO THREE WEEKS, IT'S VERY
CONVENIENT, ALTHOUGH IT DOES PUT
A HAMPER ON YOUR SCHEDULE,
VERSUS OURS WHICH IS INCREDIBLY
COMPLICATED, YOU HAVE TO HAVE AN
OPERATION, COME BACK TO THE
HOSPITAL IN THREE TO FOUR WEEKS
BUT YOU ONL HAVE TO DO IT ONCE.
SO IT A TRADEOFF THAT MANY
PATIENTS ARE FACING.
PATIENTS WITH LYMPHOMA ARE
FACING THIS AS WELL, DO YOU TAKE
THE DRUG YOU TAKE EVERY DAY OR
DO YOU GET THE EXPENSIVE CAR
THERAPY THAT REQUIRES YOU TO BE
IN THE HOSPITAL FOR TWO WEEKS?
THESE TRADEOFFS ARE GOING TO BE
WHAT DRIVES PATIENTS WITH CANCER
ARE CHOOSING IN THE FUTURE.
SO THIS SOMEWHERE W IS WHERE WE.
THE NEW PD-1 LAG 3 DRUG, THEN
THEY COME IN TO GET TIL
RESECTION, THEN THEY GET THEIR
CELL THERAPY.
WHERE WE STAND TO LEARN THE MOST
IN MY OPINION IS IF WE GET THE
OPPORTUNITY TO RESECT A TUMOR
BEFORE THEY HAVE ANY OF THOSE
THERAPIES.
SO THAT WE CAN UNDERSTAND
WHETHER THOSE THERAPIES ARE
ACTUALLY IMPACTING HOW WE GROW
THE TIL OR IMPACTING HOW THE TIL
ARE AFFE ABLE TO AFFECT THEIR CR
AFTERWARDS AND IN THE FUTURE,
DOG THE CELL THERAPY FIRST.
DOING THE CELL THERAPY FIRST.
IOVANS HAS LOOKED AT THAT AND
NOW RUNNING A TRIAL OF TIL
VERSUS PEMBROLIZUMAB AND IN
THEIR SMALL COHORT OF I BELIEVE
IT'S 20 OR SO PATIENTS, YOU CAN
GET THESE GREAT RESPONSES.
A 30% COMPLETE RESPONSE RATE TO
TIL PLUS PEMBRO IN
TREATMENT-NAIVE PATIENTS.
SO THAT IN MY OPINION IS WHERE
WE COULD BE GOING IF PATIENTS
ARE WILLING TO WORK THERE WITH
US.
SO THE EVOLVING THERAPEUTIC
LANDSCAPE OF MELANOMA, IT CANNOT
BE SAID ENOUGH THAT CHECKPOINT
BLOCKADE HAS REVOLUTIONIZED THE
CARE OF THESE PATIENTS.
IT IS RESPONSIBLE FOR A NEAR
HALVING OF THE MORTALITY RATE
OVER THE LAST FIVE YEARS IN
PATIENTS WHO HAVE METASTATIC
DISEASE.
FAST, ACCESSIBLE TUMOR
SEQUENCING HAS CHANGED OUR
THERAPIES AND RESEARCH.
WE NOW KNOW ABOUT DRIVER
MUTATIONS, WE UNDERSTAND BETTER
THE TUMOR-TIL RELATIONSHIP.
TIL CAN MEDIATE LONG TERM
DURABLE RESPONSES AND I THINK
THAT IS REALLY THE KEY.
TIL AS WE ALL KNOW IS NOW
APPROVED IN TREATMENT REFRACTORY
DISEASE, HOWEVER, PRIOR THERAPY
DOES LOWER THE LIKELIHOOD OF
THAT RESPONSE, SO IS THERE ROOM
FOR TIL AS A FRONT-LINE
STRATEGY?
BECAUSE IN MY OPINION, WE WANT
TO GET ALL OUR PATIENTS TO THIS
GREEN LINE.
THIS WAS ONE OF MY FONDEST
MEMORIES.
IT'S NOT OFTEN THAT YOU GET TO
SURPRISE DR. ROSENBERG, BUT WE
ACTUALLY MADE A WORKING
OPERATION GAME FOR OUR HOLIDAY
DOOR CONTEST A FEW YEARS BACK.
AND I KNOW ANYONE THAT'S WORKED
WITH HIM HAS HEARD HIM SAY THIS
MORE THAN ONCE, EVERY PATIENT
DESERVES AN OPTIMISTIC
ONCOLOGIST.
AND SO WITH THAT, I'LL TAKE ANY
QUESTIONS.
AND CERTAINLY WANT TO
ACKNOWLEDGE EVERYONE AT THE
BRANCH AND MOST PARTICULARLY OUR
PATIENTS AND THEIR FAMILIES.
[APPLAUSE]
I GOT OFF EASY.
NO QUESTIONS.
>> WHAT ABOUT CLASS 1 -- TUMORS?
>> WHAT ABOUT CLASS 1 LOSS
TUMORS?
YOU KNOW, WE'VE EXPLORED THAT
MORE IN OUR PATIENTS WITH
EPITHELIAL CANCER THAN WE HAVE
WITH MELANOMA, SO WE'RE STARTING
TO GO BACK AND LOOK AT THAT MORE
DEEPLY.
WHAT WE HAVE FOUND IS THAT A
SIGNIFICANT NUMBER OF OUR
PATIENTS HAVE ACTUALLY LOST
CLASS 1 AND WE NOW USE IT AS A
SCREENING BEFORE WE DO SOME OF
OUR GENE ENGINEERED THERAPIES
BECAUSE WE KNOW IT'S A MECHANISM
OF REEF CYST TANS.
WHAT HE WITH HOPE IS THAT
BECAUSE WE'RE GIVING A
POLYCLONAL PRODUCT THAT EVEN IF
THEY LOSE ONE OF THEIR ALLELES,
THEY'VE GOT THE OTHER AND
HOPEFULLY THEY'VE GOT A
POLYCLONAL POLYTARGETTED
THERAPY.
WHICH IS WHY I THINK TIL PLAY
HAVE IN MY OPINION AN ADVANTAGE
OVER SINGLE CELL THERAPIES.
>> [INAUDIBLE]
>> THEY DON'T GROW WELL IN THE
CONDITIONS, BUT THAT'S BECAUSE
WE DRIVE OUR GROWTH THAT WAY.
IF WE WANTED TO GROW
GAMMA-DELTA, WE'D HAVE TO CHANGE
THE WAY WE GROW OUR CELLS.
>> [INAUDIBLE]
>> PAUL ROBBINS, EVERYONE.
AND YOU'LL GET A CHANCE TO HEAR
FROM HIM A LITTLE BIT LATER
TODAY.
>> STEPHANIE, GREAT TALK.
I'D LIKE TO JUST FOLLOW UP ON
THIS ISSUE OF THE PD-1
REFRACTORY MELANOMAS.
DO YOU THINK THE ANTIPD-1
THERAPY IS JUST IDENTIFYING
MELANOMAS THAT ARE POORLY
IMMUNOGENIC SO THAT EVEN IF YOU
TREAT THEM IN THE FIRST LINE
WITH TIL, THEY'RE NOT GOING TO
BE GOOD RESPONDERS?
>> I THINK IT'S -- I THINK IT'S
A LITTLE BIT OF BOTH.
SO I THINK IF WE WEREN'T SEEING
ANY RESPONSES IN THE POST PD-1
PATIENTS, I WOULD THINK THAT
IT'S THAT IT'S IMMUNOEDITING TO
A POINT THAT WE CAN NO LONGER
EXPRESS IT.
THAT'S WHY I'D LIKE TO STUDY
MORE TEU NORS BEFORE PD-1 TO BE
ABLE TO COMPARE.
WE HAVE BEEN ABLE TO FIND
NEOANTIGENS IN THOSE PATIENTS,
WE JUST HAVEN'T BEEN ABLE TO
FIND THEM AS ROBUSTLY.
I KNOW THAT'S A NEW WAY WE'VE
STARTED SCREENING THEM.
ONE THING I DO WANT TO TRY TO
PUT FORWARD CLINICALLY NOW IS
NOT ONLY TAKING PART IN THE
RANDOMIZED TRIAL OF GROWING TIL
IN A STANDARD FASHION, BUT
TAKING WHAT WE'VE LEARNED FROM
EPITHELIAL AND ACTUALLY
SCREENING FOR NEOANTIGEN
REACTIVITY TO SEE IF WE CAN
ENRICH AND MAKE THE TIL FOR POST
PD-1 PATIENTS MORE EFFECTIVE.
>> THANK YOU.
>> VERY NICE TALK.
SAMIR KALIFF FOUND IN CANCER
VACCINE STUDY IN MICE THAT
PRE-TREATMENT WITH ANTIPD-1 LED
TO LESS EFFECTIVE VACCINE
RESPONSE THAN TREATING
CONCURRENTLY, AND WAS ASSOCIATED
WITH MORE CD8 -- CD38-POSITIVE
CD-8 T CELLS THAT WERE SO-CALLED
SUBPRIMED.
AND WE TESTED THAT IN OUR
VACCINE SYSTEM AND FOUND
SOMETHING VERY SIMILAR, THAT
PRE-TREATING WITH ANTIPD-1
ACTUALLY REDUCED THE
EFFECTIVENESS OR ELIMINATED THE
EFFECTIVENESS OF THE VACCINE
COMPARED TO GIVING IT
CONCURRENTLY OR AFTER THE
VACCINE.
SO I WONDER IF THAT'S RELATED AT
ALL TO WHAT YOU'RE DESCRIBING
HERE WITH THE TILs, AND HAVE
YOU HAD THE OPPORTUNITY TO LOOK
AT CD38 EXPRESSION IN THE PEOPLE
WHO ARE PRE-TREATED WITH
ANTIPD-1?
>> WHAT I WILL SAY IS IN THE
RECENT TWO TRIALS OF USING
CANCER VACCINES IN PATIENTS,
BOTH WITH MELANOMA AND WITH
PANCREAS CANCER, THOSE ARE GIVEN
IN CONJUNCTION WITH AN ANTIPD-1
OR ANTIPDL1 INHIBITOR.
SO THE PATIENTS WILL GET THE
DRUG, THEN GET THE VACCINE, AND
BOTH IN THE PAPER THAT DR. WEBER
PUBLISHED RECENTLY BEFORE HIS
PASSING AND IN THE WORK THAT
CAME OUT OF MEMORIAL SLOAN
KETTERING, THOSE PATIENTS WERE
ABLE TO BE VACCINATED.
AND SO THE EFFECT OF THAT
CHECKPOINT INHIBITOR IN HUMANS
DOES NOT SEEM TO DIMINISH THE
ABILITY TO RAISE THAT
IMMUNOGENIC RESPONSE.
ONE THING THAT WE HAVE LOOKED AT
IS NOT CD38 SPECIFICALLY, BUT
CD39 AND 69, AND I THINK RATHER
THAN STEAL THE THUNDER OF EITHER
DR. LOWRY OR DR. KRISHNA WHO
WILL PROBABLY PRESENT THAT MORE
FULLY LATER THIS AFTERNOON, WE
DID FIND THAT THOSE CELLS THAT
WERE NEGATIVE WERE LESS PRESENT
IN THE PD-1 GROUP.
>> OKAY.
THANK YOU.
>> THANK YOU ALL.
OH.
>> ONE COMMENT I'D LIKE TO MAKE
IS THAT WHEN YOU LOOK AT THE
COMPARISON BETWEEN THE VARIOUS
IMMUNOTHERAPIES AVAILABLE AND
THE -- THE -- ONE THING THAT'S
NOT MEASURED IS THE TIME OFF
TREATMENT.
THE AMOUNT OF TIME THE PATIENT
DOES NOT SEE A DOCTOR OR A
HOSPITAL OR A CLINIC.
THAT'S SOMETHING THAT WE NEED TO
CAPTURE, THE DIFFERENCE BETWEEN
THE -- PRETTY MUCH ALL OTHER
THERAPIES.
>> I AGREE, AND THE GROUP IN
EUROPE ACTUALLY INCORPORATED
QUALITY OF LIFE STUDIES IN THEIR
RANDOMIZED TRIAL AND FOUND THAT
QUALITY OF LIFE WAS BETTER IN
THE PATIENTS THAT HAD GOTTEN THE
TIL PRODUCT THAN THOSE THAT WENT
ON TO IPIMIBULAB.
THANK YOU FOR YOUR ATTENTION.
AND AGAIN, DR. ROSENBERG, I
WOULDN'T BE HERE WITHOUT YOU,
QUITE LITERALLY.
THANK YOU.
[APPLAUSE]
>> OKAY, THANK YOU, EVERYBODY.
THAT'S IT FOR OUR FIRST SESSION.
SO WE HAVE TIME FOR A 15-MINUTE
BREAK AND THEN WE'LL COME BACK
AT 9:50 FOR ANTIGEN DISCOVERY
WITH DR. PRIETO.
THANK YOU.
MORNING SO WE'RE GOING TO START
WITH OUR SECOND SESSION, WHICH
IS DEALING WITH ANTIGEN
DISCOVERY.
WE HAVE WITH US FIRST DR. YUTAKA
KAWAKAMI FROM THE INTERNATIONAL
HEALTH AND WELLNESS UNIVERSITY
FROM SHIBA, JAPAN.
DR. KAWAKAMI.
>> GOOD MORNING.
THANK YOU, CHAIRMAN AND THOSE
MEETING ORGANIZERS.
THIS IS MY GREAT HONOR FOR ME TO
GIVE A TALK AT THIS SPECIAL
SYMPOSIUM CELEBRATING
DR. ROSENBERG'S 50 YEARS AT NCI.
I STILL REMEMBER WHEN I WAS A
PHYSICIAN, HEMATOONCOLOGIST
TRAINED AT UNIVERSITY OF TOKYO,
I WAS VERY INTERESTED WITH ONE
PAPER WRITTEN BY DR. ROSENBERG
DISCUSSING POTENTIAL NEW CANCER
TREATMENT THAT WAS -- TRANSFER
OF LYMPHOCYTE.
SO I HAD A CHANCE TO JOIN THE
BRANCH.
I WAS HERE '87 TO '97, FOR 10
YEARS.
SO TODAY, I'M GOING TO TALK
ABOUT TUMOR ANTIGEN ISOLATION, A
HISTORICAL VIEW AND ALSO A
LITTLE BIT ABOUT FURTHER STUDY
AND RESEARCH.
SO THIS SLIDE SHOWS OUR CANCER
IMMUNOLOGY RESEARCH HISTORY.
SO ANTITUMOR ACTIVITY OF
CD8 POSITIVE TUMOR LYMPHOCYTE
WAS DISCOVERED AT THE BRANCH, WE
TRIED TO ISOLATE TUMOR ANTIGEN
RECOGNIZED BY -- TO UNDERSTAND
MECHANISM OF ANTITUMOR IMMUNE
RESPONSE AND THOSE MECHANISMS OF
IMMUNORESISTANCE.
SO LATER, 2010, LATER
CHECKPOINT -- WAS AVAILABLE,
COMPARATIVE ANALYSIS BETWEEN THE
RESPONDER AND NON-RESPONDER
ACTUALLY PROMOTED FURTHER CANCER
IMMUNOLOGY RESEARCH.
NOW WE KNOW MANY IMMUNOLOGICAL
SUBTYPE VARIETY OF CANCER AND
THUS A POTENTIAL GOOD
COMBINATION IMMUNOTHERAPY.
SO THIS IS A PICTURE, I WAS HERE
AT THE NIH.
SOME OF THE PEOPLE ACTUALLY HERE
TODAY.
THIS IS THE TUMOR ISOLATION,
ACTUAL MORE ANTIGEN ISOLATION
GROUP.
ALSO WE COLLABORATED WITH MANY
RESEARCHERS AT NIH AND AS AN
INSTITUTE FOR C D EXPRESSION
CLONING, BINDING PEP TIGHTS,
SPECTROMETRY ANALYSIS, NEW
TECHNOLOGY WAS ESSENTIAL AND
EVEN NOW IT'S MORE IMPORTANT TO
COLLABORATE WITH MANY PEOPLE.
SO WE FIRST ACTUALLY FOUND THREE
STRATEGIES, CLONING -- USED
FIRST STABLE EXPRESSION SYSTEM,
THEN COS CELL TRANSIENT SYSTEM.
TESTING KNOWN OR NEW CANDIDATE
IDENTIFIED BY A VARIETY OF
DIFFERENT METHODS.
THE THIRD ONE IS ISOLATION,
DIRECT ISOLATION OF HLA BINDING
PEPTIDES ON HLA MELANOMA CELL,
USING HPLC AND MASS
SPECTROMETRY.
UNFORTUNATELY I COULD NOT --
ISOLATE THIS WAY BUT LATER, THIS
IS VERY IMPORTANT TO JUST
CONFIRM THE PRESENCE OF PEPTIDE
ON HLA ON MELANOMA CELL.
WHEN I WENT BACK TO JAPAN, I
ALSO TRIED CD EXPRESSION CRO
CREXPRESSIONCLONING.
TUMOR ANTIGEN AND
CHARACTERIZATION OF ANTIGEN FOR
PREPARATION OF -- FOR
ANTISCREENING, WE'RE SCREENING
SO MANY TIL, RESULT IN TUMOR
REGRESSION.
HERE SHOW GP100 REACTIVE T-CELL
VERY GOOD RESPONSE.
WE ALSO EVALUATED -- OF
AUTOLOGOUS AND ALLOGENIC
MELANOMA CELL, ACTUALLY WE
PROVED THE PRESENCE OF SHARED --
ALSO -- BY VERY COMMON HLA-A2.1,
LATER A20, 1/3 INTSH 26,
SOMETHING LIKE THAT.
THIS WAS PRETTY NEW AT THE TIME,
IF YOU REMEMBER, THE STRUCTURE
OF HLA-ASM 26789 -- WAS JUST
PUBLISHED 1987.
TWO YEARS BEFORE I STARTED THE
TUMOR ANTIGEN PROJECT.
WE ALSO MADE SURE THE ENDODGE
JUST EXPRESSION OF EPITOPE, WE
CONFIRMED THE PRESENCE OF
EPITOPE PEPTIDE AND ALSO TUMOR
DEACTIVE T-CELL BY STIMULATION
WITH PEPTIDE.
TO CHARACTERIZATION OF TUMOR
ANTIGENS AND EP TOTALS, SELF OR
MUTATED, OR SHARED OR UNIQUE BY
DNA SEQUENCING AND ALSO HLA
BINDING AFFINITY OF PEPTIDES,
MANY OF THEM IT'S INTERMEDIATE
AFFINITY.
WE CAN MODIFY TO MAKE
IMMUNOGENIC PEPTIDE, AND ALSO
THE AVIDITY OF TCR TO
PEPTIDE/HLA COMPLEX, WE CAN MAKE
A HIGH AVIDITY TCR.
AND BIOLOGICAL FUNCTION OF
ANTIGEN, THIS IS ALSO IMPORTANT
RELATED TO THE UNDERSTANDING OF
IMMUNOLOGIST -- ME CANNISTS AND
TARGET THERAPY.
OF COURSE INVIE VO ANTITUMOR
ACTIVITY WAS CONFIRMED IN
CLINICAL TRIAL.
NOW MANY PEOPLE TRYING TO
GENERATE ALGORITHM PREDICTING I
MU KNOWGENIC NEOANTIGEN, ALSO
MAKING A LIBRARY OF TUMOR
ANTIGEN AND THEY'RE RECOGNIZING
TCR.
NOW THIS IS WHERE I JUST SHOW
REPRESENTATIVE -- WE FIRST
ACTUALLY ISOLATED TISSUE --
ANTIGEN.
LATER THE NEOANTIGEN DID WITH
MYSELF ISOLATED SINGLE --
PINPOINT MUTATION.
THIS MUTATION CREATES HIGHER HLA
BINDING PEPTIDE, WHICH ACTUALLY
INDUCED T-CELL.
WE ALSO ISOLATE -- VERY SIMILAR
MECHANISM.
THEN LATER WE ALSO FOUND IN
COLON CANCER -- DEFECTIVE ONE,
WE FOUND THE IMMUNE SYSTEM
ACTUALLY RECOGNIZED -- PEPTIDE.
THIS IS VERY IMPORTANT TOO.
NOW, THIS IS -- I MADE LATE NIEP
LATE90s SHOWING -- ON CANCER CE.
TRANSLATION, MODIFICATION,
VARIETY OF MECHANISMS TO
GENERATE T-CELL EPITOPE ON
CANCER CELL.
I WILL NOT GET IN DETAIL.
THIS IS ALSO LATE N LATE '90S, G
HLA PEPTIDE, AFFINITY MEANS --
CRYPTIC NATURE OF EPITOPE, WHICH
MEANS MAYBE WE CAN MAKE A
MODIFICATION.
AND ALSO WE MADE SURE -- PRESENT
OF THIS ANTIGEN OF -- MELANOMA.
SHOWING TARGET USED FOR CLINICAL
TRIAL.
-- T-CELL, THIS PEPTIDE, THIS
PEPTIDE MODIFIED GP100 USED FOR
VACCINE TRIAL.
THIS PEPTIDE IS ACTUALLY
BIOSTATISTIC -- CD3 AND THIS
PEPTIDE -- WAS ACTUALLY APPROVED
BY FDA.
THIS IS ACTUALLY EXPERIMENT OF
CANCER VACCINE WITH HIGHLY I YOU
MOO KNOWGENIC MODIFIED GP100
PEPTIDE.
THE SERIES OF STUDIES SHOW
ANTI-TUMOR ACTIVITY OF SHARED --
OR WE CAN SEE THE -- BUT ANYWAY,
IT IS A VERY NICE -- TUMOR
ANTIGEN MADE AS POSSIBLE TO
DEVELOP ANTIGEN-SPECIFIC
IMMUNOTHERAPY IN A VERY
SCIENTIFIC WAY.
THIS IS WHAT'S GOOD.
AND I THINK STILL THAT EVEN
THE -- T-CELL IN MELANOMA NEED
TO BE CLARIFIED.
LIKE VACCINE STUDY AND ALSO
TRANSFER OF MART 1 -- TRANSDUCED
T-CELL AND ALSO OLD PAPER BUT --
CORRELATE RESPONSE TO
INTERLEUKIN 2 THERAPY AND ALSO
RECENTLY THIS IS JEFF WEBER'S
PAPERS IN RESPONSE TO PD-1
TREATMENT.
SO MAYBE THERE WILL BE SOME LOAD
OVER -- ANTIGEN SPECIFIC T-CELL.
THIS IS A VERY INTERESTING
CLINICAL HISTORY ON A PATIENT
TREATED WITH -- SO ONE THING
INTERESTING -- TIL THERAPY --
NEOANTIGEN WAS VERY EFFECTIVE
DND MULTIPLE METASTASES.
-- HLA LOSS OCCUR.
PREVIOUSLY WE REPORTED IN THE
INTERLEUKIN 2 THERAPY.
AND INTERESTINGLY -- CANCER CELL
STILL EXPRESSING NEOANTIGEN
REMAINING EVEN IN COMPLETE
RESPONSE PATIENT.
THEN THREE YEARS LATER, TWICE,
THAT MEANS -- IMMUNORESISTANT
DOMINANT CANCER CELERY MAINING.
THEN THAT LED US ANALYSIS OF
CANCER STEM CELL EMT, LATER WE
FOUNT EMT CANCER CELL INCREASED
METASTASES PARTLY DUE TO VERY
STRONG IMMUNOSUPPRESSION.
NOW, THIS IS A PROTOTYPE OF
IMMUNOLOGICAL SUBTYPE IN
MELANOMA.
AS YOU ALREADY KNOW,
HYPERMUTATION TYPE -- AND
VARIETY OF IMMUNOSUPPRESSION.
ACTIVATED ONCO -- MUTATION --
RELATED TO EMT, CAF, ALTERED
METABOLISM, WE'RE NOW LOOKING ON
FATTY ACID, CHOLESTEROL AND ALSO
GUT MICROBIOTA -- MEDICINE.
NOW, THIS IS ONE -- OUR
RESEARCH -- DENDRITIC CELL
T-CELL ACCESS AFTER -- MUTATION
WAS DISCOVERED WE ACTUALLY WORK
ON BETA -- INDUCE
IMMUNOSUPPRESSION.
DIFFERENT MECHANISM THAN
INHIBITOR ACTIVATE ANTITUMOR
DENDRITIC CELLS -- NOW
DOWNSTREAM OF THE -- IS CD1,
THIS IS A FATTY ACID --
INHIBITOR ACTIVATE TUMOR --
AGAIN 16ER J SYNERGIZE WITH PD-.
THEN WE ARE FOCUSING INTRATUMOR
DENDRITIC CELL.
JIM MULE DISCUSSED ABOUT -- IN
CANCER, MANY PEOPLE WORKING ON
THAT.
NOW, WE ACTUALLY ARE WORKING
ON -- DENDRITIC CELL,
MICROBIOTA -- INHIBITOR -- THOSE
ACTUALLY ACTIVATE INTERTUMOR
DENDRITIC CELLS AND FINALLY
ACTIVATE T-CELL -- PD-1.
THIS IS ONE EXAMPLE WE
PREVIOUSLY IDENTIFIED --
IMMUNOSUPPRESSION.
THIS IN MOUSE -- MELANOMA,
INHIBITOR ANTIPD-1 ANTIBODY IS
EFFECTIVE BUT NOT ENOUGH.
IN ADDITION OF -- AGONIST IS
COMPLETELY ERADICATE ACTIVATION
OF INTRATUMOR DENDRITIC CELL,
CDC, PDC -- THIS WAS USED --
SIMILAR 2003.
NOW ACTUALLY INTERESTING
SITUATION, IL-2 RESPONSE
CANCER -- DEFICIENT CANCER,
VIRAL ASSOCIATED CANCER.
SO ONE IS BACK TO 2003, ACTUALLY
WE ANALYZED COLON CANCER WITH --
DEFECTIVE COLON CANCER -- T-CELL
INFILTRATION.
WE COULD REPORT IMMUNE
RECOGNITION AGAINST -- PEPTIDE,
THIS IS COMPLETELY CHANGED
BECAUSE OF DNA -- BY -- THEN
ACTUALLY CANCER RESEARCH -- WITH
DEFECTIVE -- IT TYPE OF TUMOR IS
PARTICULARLY INTERESTING IN
TERMS OF -- VIEW.
THEN LATER ACTUALLY PD-1
ANTIBODY WAS FOUND TO BE VERY
EFFECTIVE FOR -- CANCER,
ALTHOUGH SOME OF THEM STILL DO
NOT RESPOND SO -- CLASS 1 LOSS
OR IMMUNOSUPPRESSION -- TO ONE
OF THE MECHANISMS OF
SUPPRESSION.
NOW IN TERMS OF -- RECOGNIZE --
TUMOR ORGANOID AND THOSE ARE
FREQUENTLY -- HBV67.
CLINICAL TRIAL -- SIMILAR TO THE
PROTOCOL ACTUALLY -- PATIENT WAS
ALMOST COMPLETE RESPONSE.
NOW, THIS IS SHOWING -- I TOLD
YOU I WAS SO AMAZED BUT --
PROTEIN WHICH RECOGNIZED CD3 AND
GP100 PEPTIDE -- THIS ONE WAS
ACTUALLY APPROVED BY FDA FOR --
MELANOMA.
SO WE ALSO IDENTIFY MANY
ANTIGEN, ONE IS ONCOANTIGEN,
VERY EFFECTIVE PARTICULARLY IN
COMBINATION WITH ANTIPD-1 OR --
WE NOW TRY TO DEVELOP A CLINICAL
TRIAL.
SO THIS IS A SUMMARY OF ANTIGEN.
I'M NOT GOING TO -- THE ANTIGEN
ISOLATION, SCIENTIFIC -- T-CELL
NEO -- IS VERY IMPORTANT AND --
TARGETING TUMOR ANTIGEN IS VERY
PROMISING.
SO THIS IS -- MY COLLEAGUES, AND
TUMOR ISOLATION TEAM.
THANK YOU VERY MUCH.
[APPLAUSE]
>> ARE THERE ANY QUESTIONS FOR
DR. KAWAKAMI BEFORE WE MOVE ON
TO OUR NECK SPEAKER?
OKAY.
THE NEXT SPEAKER IS DR. PAUL
ROBBINS FROM THE CENTER FOR
CANCER RESEARCH AND MEMBER OF
THE SURGERY BRANCH.
>> ALL RIGHT.
I'M VERY HAPPY TO PRESENT TODAY.
ON THE USE OF PERSONALIZED
T-CELL ANTIGEN SCREENING
APPROACHES TO DEVELOP EFFECTIVE
ADOPTIVE IMMUNOTHERAPIES.
NOW, I WANTED TO START WITH THIS
SLIDE, WHERE I SHOW AN IMAGE OF
A CHARACTER IN GREEK MYTHOLOGY
AS EVERYBODY KNOWS, WHO WAS
PUNISHED BY THE GODS FOR KILLING
VISITORS ACTUALLY, BY BEING
FORCED FOR ETERNITY TO ROLL A
ROCK UP A HILL THAT THEN RODE
BACK DOWN AGAIN.
HIS WIFE GAVE HIM THIS FIGURE I
THINK WHEN HE FIRST CAME HERE.
APART FROM THE OBVIOUS THAT
STEVE HAS WELCOME AND CARED FOR
MORE PEOPLE THROUGH HIS WORK
THAN ANYBODY LISTENING IN THE
AUDIENCE, I BELIEVE THIS IMAGE
ALSO REALLY DOESN'T CAPTURE HIM.
AND THIS IS, I THINK, EVIDENT
HERE.
IF YOU LOOK AT PROGRESS IN
IMMUNOTHERAPY, OVER MEAN YEARS,
I THINK STEVE HAS REALLY PLAYED
A MAJOR ROLE IN ADVANCING
THERAPY.
SO THIS IS REALLY THE IMAGE THAT
I HAVE WHEN THINKING OF HIM.
HE'S ROLLING THE ROCK UP THE
HILL, WE HAVEN'T REACHED THE
TOP, WE'RE STILL ON OUR WAY,
STILL MAKING PROGRESS BUT I
THINK HE'S PLAYED A MAJOR ROLE
IN ROLLING THAT ROCK UP THE
HILL.
YOU CAN SEE HIS NAMEPLATE HERE
NEXT TO THIS FIGURE.
SO I WANT TO DISCUSS HERE WHAT
SOME OF THE MAIN ISSUES FOR THE
DEVELOPMENT OF EFFECTIVE
ADOPTIVE T-CELL THERAPIES.
I THINK UNDERSTANDING THE
FACTORS ASSOCIATED WITH THE
RESPONSE IS CRITICAL.
THE NATURE OF THE FUNCTION OF
THE ANTIGEN TARGET WHICH IS WHAT
I'M REALLY GOING TO FOCUS ON,
EXPRESSION, MHC BINDING, THE
CLASS OF T CELLS, YOU CAN
PROBABLY COME UP WITH A VERY
LONG LIST OF THINGS THAT ARE
IMPORTANT FOR THERAPY.
DEVELOPING STRATEGIES FOR
OVERCOMING TUMOR RESISTANCE.
I THINK WE NEED TO BROADEN THE
REP TRREPERTOIRE OF ANTIGENS TOL
WITH ANTIGEN AND/OR MHC LOSS.
AS I MENTIONED, WE DON'T SEE
COMPLETE LOSS OF MHC CLASS 1 IN
GI CANCERS, AT LEAST IN THE
COHORT OF PATIENTS THAT WE'VE
LOOKED AT.
IT'S VERY RARE, IN FACT, I CAN'T
EVEN RECALL A PATIENT WHO HAS
HAD A BA BETA 2 MICRO GLOBULIN T
RESULTS IN LOSS.
BUT I THINK WE NEED TO FOCUS ON
DRIVER MUTATIONS, SINCE THESE
WILL BE ESSENTIAL FOR THE
PHENOTYPE OF THE TUMOR.
I'M NOT GOING TO TALK ABOUT
EVALUATING T-CELL STATES, BUT
SEVERAL OTHER SPEAKERS ARE GOING
TO TALK ABOUT THAT.
NOW THE EXPRESSION CLONING
SYSTEM THAT I AND YUTAKA WORKED
ON WAS FIRST ACTUALLY DESCRIBED
IN 1987 WH 1987 BY BRIAN SEED, D
FROM HIS APPROACH.
INTERESTINGLY ENOUGH THE
MOLECULE HE FIRST DISCOVERED WAS
CD28, SO A VERY RELEVANT
MOLECULE TO A LOT OF THE STUDIES
THAT WE'VE DONE.
I DON'T NEED TO GO THROUGH THIS
SLIDE IN DETAIL, I THINK, BUT
BASICALLY EVERYBODY PROBABLY IS
FAMILIAR WITH THIS.
WE ISOLATE RNA FROM TUMOR CELLS,
MAKE A CDA LIBRARY, GENERATE
POOLS OF BARACK TI IN BACTERIA,N
TRANSFECT THEM AND ISOLATE THOSE
CLONES FROM THOSE CELLS.
THIS LED TO THE IDENTIFICATION
OF MULTIPLE CATEGORIES OF
ANTIGENS.
YUTAKA WENT OVER THESE IN MORE
DETAIL.
BUT ONE OF THE IMPORTANT
PRODUCTS THAT WERE IDENTIFIED
EARLY ON ARE THE NEOANTIGENS,
THE PRODUCT OF MUTATED GENES.
NOW THE AGNOSTIC NATURE OF THIS
CLONING SYSTEM REALLY LED TO
IDENTIFICATION OF SOME
NON-CANONICAL PRODUCTS AS WELL.
AGAIN, YUTAKA MENTIONED SOME OF
THESE TOO.
BUT I LIKE TO THINK OF THESE AS
THE DARK MATTER OF THE IMMUNE
SYSTEM, WHICH IS A POPULAR TERM
NOW OBVIOUSLY.
THE PEPTIDE SPICING WAS SHOWN
FIRST ACTUALLY BY KEN, WHERE
THEY'RE SPLICED POST
TRANSLATIONALLY IN THE
PROTEASOME, SUBSEQUENTLY SHOWN
THAT THIS OCCURS IN THE
PROTEASOME, AND THESE PEPTIDES
CAN BE SPLICED IN EITHER ORDER.
SO THE END TERMINAL PART OF THE
PEPTIDE CAN BE AT THE N-TERMINUS
OF THE PROTEIN OR AS THE
C-TERMINUS AS YOU CAN SEE BY THE
ARROW HOW THESE PEPTIDES ARE
JOINED.
EPITOPES ARE ALSO GENERATED BY
THE TRANSLATION OF ALTERNATIVE
OPEN READING FRAMES.
AS YOU CAN SEE HERE, THE
SEQUENCE SHOWN BELOW.
YOU HAVE TWO ALTERNATIVE OPEN
READING FRAMES DOWNSTREAM OPEN
READING FRAME THAT IS USED TO
INITIATE THE PRODUCTION OF A
PROTEIN THAT'S THEN PROCESSED
INTO A PEPTIDE.
THEN EPITOPES WITHIN INTRONS,
EITHER PARTIALLY OR ENTIRELY
WITH INTRONS.
ACTUALLY THE FIRST MUTANT TUMOR
ANTIGEN THAT WAS IDENTIFIED IN
HUMANS, FIRST CALLED MUM1 WAS
ACTUALLY THE PRODUCT OF AN
INTRON, WAS TRANSLATED FROM AN
INTRON.
NOW, THE ADVANTAGES OF THE CDNA
LIBRARY SCREENING APPROACH I
THINK, THOUGH, NEED TO BE
BALANCED BY THE LIMITATIONS.
ANTIGENS WERE READILY IDENTIFIED
IN THIS SYSTEM BECAUSE THEY WERE
HIGHLY REPRESENTED, HIGHLY
EXPRESSED IN MOST OF THE CELL
LINES AND A, THEREFORE, IN THE
LIBRARIES THAT WE MADE.
SO WE INITIALLY FOCUSED ON
MDAs AS TARGETS FOR THERAPY,
BUT IN TRIALS CARRIED OUT USING
TCRs DIRECTED AGAINST MAR1,
GP100, WE SAW DURABLE RESPONSES
IN ONLY A VERY SMALL MINORITY OF
PATIENTS.
BUT I THINK ANOTHER IMPORTANT
POINT IS THAT THE NEED FOR CELL
LINES AND LENGTHY PROCEDURES
INVOLVED WITH SCREENING MAKE
THIS ROUTINE SCREENING DIFFICULT
TO CARRY OUT.
CERTAINLY NOT PROSPECTIVELY ON A
PATIENT WHO WE WISH TO TREAT,
BECAUSE THIS CAN TAKE --
GENERALLY TAKES MANY MONTHS TO
CARRY OUT.
NOW, THE DRAMATIC DECLINE IN
SEQUENCING COSTS, THOUGH, IN THE
LATE TWO THOUSANDS, LED TO THE
DEVELOPMENT OF A NEW ANTIGEN
DISCOVERY PLATFORM, AND I'D LIKE
TO SHOW THIS BECAUSE IT SHOWS
YOU THE COST PER MEGABASE OF DNA
SEQUENCING AND THE DECLINE OF
THAT COST PARTICULARLY OVER THE
2000s.
YOU CAN SEE MOORE'S LAW PLOTTED
HERE, WHICH BASICALLY WAS
DEVELOPED BY GORDON MOORE, I
THINK BACK IN 1965, WHERE HE
PREDICTED THAT THE CAPACITY OF
CHIPS WOULD INCREASE BY TWOFOLD
EVERY TWO YEARS BASICALLY.
AND AMAZINGLY ENOUGH, MOORE'S
LAW HAS HELD FOR A LONG TIME.
I THINK FOR MANY DECADES.
BUT WHAT YOU CAN SEE HERE IS
THAT THERE WAS A PRECIPITOUS
DROP IN THE COST OF SEQUENCING
IN THE LATE 2000s.
SO OBVIOUSLY OTHER THINGS WERE
INVOLVED.
OBVIOUSLY COMPUTING TECHNOLOGY
IMPROVED, BUT THEN ADVANCED --
OTHER ADVANCED TECHNOLOGIES
ALLOWED I THE SEQUENCING COST TO
DECLINE QUITE PRECIPITOUSLY, AND
THIS ALLOWED US TO CARRY OUT A
VARIETY OF STUDIES AND OTHERS
BEGAN TO LOOK AT TUMORS, TUMOR
MUTATIONAL BURDEN, AND IN THIS
PAPER PUBLISHED IN 2017, WHICH I
THINK IS A LANDMARK PAPER
BASICALLY THIS GROUP SHOWED THAT
THE TUMOR MUTATIONAL BURDEN
CORRELATED WITH OBJECTIVE
RESPONSE TO PD-1 DIRECTED
THERAPY.
YOU CAN SEE MELANOMA OVER HERE
IS OBVIOUSLY HIGHLY RESPONSIVE
TO PD-1.
BUT AS WELL AS MISMATCH REPAIR
DEFICIENT COLORECTAL CANCER.
HOWEVER, MISMATCH REPAIR STABLE
COLORECTAL CANCER DOES NOT
RESPOND TO CHECKPOINT THERAPY AT
ANY APPRECIABLE LEVEL.
SO THE EXPRESSION CLONING SYSTEM
THAT WAS DEVELOPED BY OUR GROUP
AND THE GROUP IN BRUSSELS AS
WELL AS SEVERAL OTHER GROUPS WAS
BASICALLY A MODIFIED VERSION OF
THE EXPRESSION CLONING SYSTEM,
SO GENOMIC DNA IS ISOLATED, RNA
AS WELL FROM THE TUMOR,
GENOMIC -- IS -- WHOLE EXOME
SEQUENCE SOMETHING CARRIED OUT,
SOMATIC MUTATIONS ARE IDENTIFIED
AND THEN YOU CAN GENERATE MINI
GENES, TANDEM MINI GENES OR
PEPTIDES CORRESPONDING TO THESE
MUTANT TRANSCRIPTS, TRANSFECT
THEM OR PULSE THEM ON TO
AUTOLOGOUS DENDRITIC CELLS,
CO-CULTURE THOSE CELLS AND
IDENTIFY THE REACTIVE T CELLS OR
SHOWN ON THE RIGHT, USING AN
ALTERNATIVE PROCESS, IDENTIFY
THE T CELLS AND THE T-CELL
RECEPTORS THAT MEDIATE
RECOGNITION OF THOSE
NEOANTIGENS.
NOW NEOANTIGEN REACTIVITY HAS
CLEARLY BEEN ASSOCIATED WITH THE
RESPONSE TO ADOPTIVE
IMMUNOTHERAPY.
IN THIS PAPER PUBLISHED FROM THE
DANISH GROUP, THE FREQUENCY OF
NEW ANTIGEN-REACTIVE T CELLS
HERE IDENTIFIED USING
BARCODED -- WAS CLEARLY
ASSOCIATED WITH THE RESPONSE TO
MELANOMA TIL.
AND THEN IN OUR STUDY, WHICH WAS
PUBLISHED THE SAME YEAR, LEVI IN
OUR GROUP LOOKED AT PD-1 NAIVE
AND REFRACTORY MELANOMA PATIENTS
WHO RECEIVED ADOPTIVE CELL
TRANSFER, AND AS YOU CAN SEE
HERE, THE COMPLETE RESPONSE RATE
DROPPED PRECIPITOUSLY WHEN WE
LOOKED AT THE PD-1 REFRACTORY
PATIENTS.
INTERESTINGLY ENOUGH, THE
PARTIAL RESPONSE RATE WAS
SIMILAR IN THESE GROUPS.
BUT WHAT WAS MOST STRIKING, AND
I BELIEVE YUTAKA -- THIS SLIDE
HAS ALREADY BEEN SHOWN OR A
SIMILAR ONE -- RESPONDERS IN
BOTH THE PD-1 NAIVE AND
REFRACTORY GROUPS HAD A HIGHER
TUMOR MUTATIONAL BURDEN.
YOU CAN SEE PARTICULARLY ON THE
LEFT SIDE WHERE THERE ARE
MELANOMA TUMORS THAT HAVE VERY
HIGH MUTATIONAL BURDENS THAT THE
RESPONSE RATE, COMPLETE RESPONSE
RATE SHOWN IN THE BLUE DOT, IS
VERY HIGH, ESSENTIALLY ALL THE
PATIENTS THAT HAVE HIGH
MUTATIONAL BURDENS ARE
RESPONDING TO THERAPY.
THIS WAS ALSO TRUE FOR THE PD-1
REFRACTORY PATIENTS.
EVEN THOUGH THERE ISN'T A
SIGNIFICANCE DIFFERENCE,
STATISTICALLY BETWEEN THE NUMBER
OF MUTATIONS, WHEN YOU LOOK AT
THE RESPONDERS IN BOTH GROUPS,
YOU CAN SEE THAT WHAT'S MISSING
FROM THE PD-1 REFRACTORY
PATIENTS WITH THE EXCEPTION OF
THAT ONE PATIENT, ONE PARTIAL
RESPONDER IS THE TUMORS THAT
HAVE HIGH MUTATIONAL BURDENS.
NOW THE NUMBER OF MUTATIONS, THE
NUMBER OF NEOANTIGENS IDENTIFIED
IN BOTH OF THESE GROUPS, THOUGH,
WAS SIGNIFICANTLY HIGHER IN THE
RESPONDERS THAN THE
NON-RESPONDERS.
BUT ONE THING TO MENTION IS THAT
WHEN YOU LOOK AT THE DURATION OF
RESPONSES, THIS IS PARTICULARLY
TRUE FOR THE PD-1 REFRACTORY
PATIENT, THE PRs AS
ESSENTIALLY ALL OF THEM WITH THE
EXCEPTION OF ONE PATIENT WERE
PARTIAL RESPONDERS, WAS LOWER
THAN IN THE PD-1 NAIVE GROUP.
56% IN THE NAIVE GROUP VERSUS
ONLY 18%.
SO THE RESPONSES THAT WE DID SEE
IN THE REFRACTORY GROUP WERE OF
MORE LIMITED DURATION.
SO I THINK THIS EMPHASIZES THE
POINT AND RAISES THE QUESTION OF
WHAT'S HAPPENING IN THOSE
PATIENTS.
THERE IS LOWER MUTATIONAL BURDEN
IN THE REFRACTORY PATIENTS.
THAT CONTRIBUTES, I THINK, TO
SOME EXTENT BUT THERE MAY BE
OTHER REASONS WHY REFRACTORY
PATIENTS RESPOND AT A LOWER
RATE.
AND I THINK THAT'S SOMETHING WE
NEED TO UNDERSTAND.
NOW RECENT ANALYSIS CARRIED OUT
IN GI CANCER PATIENTS RECEIVING
TIL ACT PLUS PD-1 HAVE SHOWN
OBJECTIVE RESPONSE RATE OF
ALMOST 24%.
IN A PREVIOUS STUDY CARRIED OUT
IN OUR GROUP, 83% OF TIL FROM
G.I. PATIENTS RECOGNIZED ONE OR
MORE NEOANTIGENS.
NOW, THESE TIL WERE SELECTED
PRIMARILY FOR NEOANTIGEN
REACTIVITY AND GIVEN
PEMBROLIZUMAB.
THE RESPONSES IN THIS COHORT OF
PATIENTS WAS ASSOCIATED WITH A
NUMBER OF NEOANTIGENS
RECOGNIZED, ALTHOUGH THE NUMBER
OF TARGETS IS LOW SO THESE
DIFFERENCES MAY NOT APPEAR VERY
IMPRESSIVE, BUT I THINK WHAT'S
MORE IMPRESSIVE IS THE GRAPH ON
THE RIGHT, WHERE YOU LOOK AT THE
TREND ANALYSIS, WHERE WHEN YOU
HAVE T CELLS THAT RECOGNIZE MORE
THAN ONE TARGET, YOUR LIKELIHOOD
OF RESPONDING INCREASES.
SO THIS WAS SIGNIFICANT BY A
TREND ANALYSIS.
NOW THIS IS UNPUBLISHED AND
DR. ROSENBERG WILL BE SHOWING
SOME MORE INFORMATION ABOUT THIS
TRIAL.
BUT ONE THING THAT I WAS CURIOUS
ABOUT WAS THE COMPARISON BETWEEN
GI AND MELANOMA.
BECAUSE OF COURSE MELANOMAS HAVE
A MUCH HIGHER TUMOR MUTATIONAL
BURDEN ON AVERAGE, BUT IF YOU
SIMPLY REMOVE THOSE PATIENTS
THAT HAVE A HIGHER MUTATIONAL
BURDEN, NOW ESSENTIALLY TRY TO
NORMALIZE THE NUMBER OF
MUTATIONS PRESENT ON THOSE
CANCERS, BECAUSE THERE IS A
LARGE OVERLAP BETWEEN GI AND
MELANOMA, IT'S JUST THAT THE GI
PATIENTS, THE MICRO SATELLITE
STABLE PATIENTS DON'T HAVE
TUMORS THAT HAVE SUCH A HIGH
BURDEN, YOU SEE THAT THE --
THERE'S A SIMILAR OBJECTIVE
RESPONSE RATE.
AGAIN, 24% IN G.I. AND 29% IN
MELANOMA.
BUT WHAT YOU CAN SEE IS THE
COMPLETE RESPONSE RATE IN THE
MELANOMA COHORT WAS 19%.
SO THIS IS ONLY SLIGHTLY LOWER
THAN THE COMPLETE RESPONSE RATE
OVERALL.
WHICH I THOUGHT WAS INTERESTING.
BUT THE MAJOR DIFFERENCE IS THAT
IN THE G.I. COHORT, WHILE THERE
WERE TWO RESPONDERS THAT HAD A
DURABLE RESPONSE OF 42 MONTHS
AND THEN ONGOING RESPONSE OF 17
MONTHS IN ANOTHER PATIENT, THERE
WERE NO CRs IN THIS GROUP.
THIS IS NOT DUE TO A DIFFERENCE
IN THE RATE OF NEOANTIGEN
RECOGNITION, BECAUSE IT'S JUST A
LITTLE UNDER 2%, IF YOU LOOK AT
THE NUMBER OF MUTATIONS VERSUS
THE NUMBER OF NEOANTIGENS WE
WERE ABLE TO IDENTIFY FOR BOTH
GROUPS.
SO THAT'S NOT RESPONSIBLE FOR
THIS DIFFERENCE.
SO WHAT POSSIBLE FACTORS MIGHT
AFFECT THIS?
CERTAINLY THE NUMBER OF
NON-SYNONYMOUS MUTATIONS HAS A
ROLE.
WE HAVE A LOWER NUMBER AND
THERE'S LESS LIKELIHOOD OF
GENERATING A NEOANTIGEN
RESPONSE, BUT CERTAINLY TUMOR
HETEROGENEITY IS ANOTHER FACTOR
THAT I THINK REALLY CLEARLY
DISTINGUISHES THESE IN THE FAR
RIGHT, YOU CAN SEE MELANOMA
PATIENTS, AND THIS TREE DIAGRAM
BASICALLY SHOWS YOU THE NUMBER
OF SHARED VARIANTS THAT ARE SEEN
WHEN YOU SEQUENCE MULTIPLE
TUMORS, AND THEN THE THIRD FROM
THE RIGHT, YOU CAN SEE
COLORECTAL CANCER.
YOU CAN SEE THIS BRANCH
STRUCTURE HERE WHERE YOU SEE
MANY MORE MUTATIONS THAT ARE
SUBCLONE OR THAT ARE NOT SHARED.
CERTAINLY DIFFERENCES IN TUMOR
CELL BIOLOGY COULD ALSO ACCOUNT
FOR THIS.
AND THE FACT THAT THE MAJORITY
OF GI CANCER PATIENTS RECEIVE
MULTIPLE LINES OF CHEMOTHERAPY
WHICH ARE NOT GENERALLY
ADMINISTERED TO MELANOMA
PATIENTS.
THESE DAYS COULD ALSO
GIVE RISE TO THIS HETEROGENEITY.
SO I THINK THAT THIS IS GOING TO
BE A MAJOR CONTRIBUTOR TO THE
DIFFERENCES IN RESPONSES.
SO WHAT ARE SOME OF THE FUTURE
OPPORTUNITIES AND CHALLENGES?
I THINK WE NEED TO PRIORITIZE
TARGETS, AND I'M NOT GOING TO
TALK ABOUT IT ABOUT ABOUT TCR ,
TARGETING NEOAT GENERALS USING
T-CELL RECEPTORS.
I ANY TIME WHATTED TO POINT OUT
THIS RECENT SITE, THE CAP SER
EPITOPE DATABASE AND ANALYSIS
RESOURCE OR SE DA CEDAR DEVELOPN
ASSOCIATION WITH THE IEDB WHICH
OF COURSE HAS A WEBSITE AS WELL
THAT CATEGORIZES ANTIGENS.
THIS SITE IS COMPLETELY FOCUSED
ON CANCER EPITOPES, AND THERE'S
OVER 6300 T-CELL EPITOPES THAT
HAVE BEEN IDENTIFIED.
SO WHEN WE'RE LOOKING FOR MORE
TARGETS, THERE ARE A LOT OF
TARGETS THAT HAVE BEEN
IDENTIFIED BUT HOW DO WE
PRIORITIZE THEM?
I THINK THAT'S GOING TO BE A KEY
QUESTION.
I THINK AN OBVIOUS SET OF
TARGETS, THOUGH, ARE THE DRIVER
MUTATIONS, OR THOSE EPITOPES
THAT ARE ENCODED BY DRIVER
MUTATIONS SINCE THOSE APPEAR TO
BE CRITICAL FOR THE FUNCTION OF
THE TUMOR.
SO HERE'S JUST A PARTIAL LIST OF
SOME OF THE T-CELL RECEPTORS
THAT HAVE BEEN IDENTIFIED
AGAINST MUTATIONS IN KRAS.
P53, PIC3CA, EGFR, BRAF, NRAS,
CDKN2A, AND ACTUALLY RECENTLY A
CLASS 2 EPITOPE HAS BEEN
IDENTIFIED WHICH ISN'T ON THIS
SLIDE FOR THE P114L MUTATION.
SO THIS SLIDE IS CONTINUALLY
BEING UPDATED AS WE IDENTIFY
MORE T CELLS.
IN SOME CASES WE IDENTIFY T
CELLS AS CLASS 1 AND A CLASS
2 RESTRICTED T-CELL AGAINST THE
SAME EPITOPE.
SO I THINK THIS EMPHASIZES THE
POINT THAT THERE ARE CERTAIN
REGIONS OF PROTEINS THAT MAY BE
PREFERENTIALLY PROCESS AND
PRESENTED AND RECOGNIZED BY THE
IMMUNE SYSTEM.
SO I WANT TO SORT OF END ON
COMING BACK TO THIS WILL THIS CF
DARK MATTER, WE HAVE SO-CALLED
ORPHAN TCRs, WE'VE NOW
IDENTIFIED OVER 50 TCRs FROM
13 PATIENTS THAT RECOGNIZE THE
AUTOLOGOUS TUMOR CELL LINE BUT
FAIL TO RECOGNIZE ANY OF THE
SCREENED NEOANTIGENS.
SO WHAT CATEGORY OF ANTIGEN DOES
THERE'S T CELLS BELONG TO?
THIS COULD BE NEOANTIGENS THAT
WE HAVEN'T IDENTIFIED BECAUSE OF
LIMITATIONS IN THE TUMOR
SEQUENCING.
WE ONLY SEQUENCE A SMALL REGION
OF THE TUMOR, SO PERHAPS WE
HAVEN'T CAPTURED ALL THE
MUTATIONS THAT ARE PRESENT IN
THAT TUMOR OR THERE MAY BE OTHER
REASONS DUE TO THE DEPTH OF
SEQUENCING BEING INADEQUATE FOR
SOME REGIONS OF THE EXOME.
THESE COULD REPRESENT CANCER
GERMLINE OR TISSUE-SPECIFIC
ANTIGENS, AND WE'RE BEGINNING TO
SCREEN, WE'VE ALREADY DONE SOME
SCREENS LOOKING AT THE
OVEREXPRESSED GENE PRODUCTS THAT
ARE PRESENT ON THOSE CELL LINES
BECAUSE WE HAVE THE
TRANSCRIPTOME DATA.
BUT WE SO FAR HAVEN'T IDENTIFIED
THE PRODUCTS.
-- TO TRY TO IDENTIFY THESE
BECAUSE THAT'S AN AGNOSTIC
METHOD.
SO WE'LL HAVE TO WAIT AND SEE.
IT COULD BE THAT THIS IS MORE
DARK MATTER, THOUGH.
THESE ARE THINGS THAT ARE FROM
PARTS OF THE GENOME THAT ARE
CHARACTERIZED AS BEING
NON-TRANSCRIBED.
WE CERTAINLY KNOW THERE'S TRANS
POE SAL BIOELEMENTS.
THERE'S LOTS OF OTHER MATERIAL
OUT THERE THAT COULD GIVE RISE
TO THESE EPITOPES.
BUT I THINK THIS IS IMPORTANT
BECAUSE WE KNOW THOSE T CELLS
ACTUALLY RECOGNIZE THE CELL LINE
FROM THAT PATIENT.
SO WE WANT TO TARGET THOSE
ANTIGENS, RIGHT?
NOT EVERY NEOTOAP REACTIVE
T-CELL WILL RECOGNIZE AN
ORGANOID.
SO INTERESTINGLY, WE'RE LOOKING
AT TUMOR RESISTANCE MECHANISMS
IN THOSE CELL LINES BECAUSE WE
DON'T KNOW IN MANY CASES WHAT'S
RESPONSIBLE FOR THE LACK OF
RECOGNITION.
THEN FINALLY I THINK WE NEED TO
USE MACHINE LEARNING APPROACHES.
WE NEED TO UNDERSTAND SOMETHING
ABOUT THESE ANTIGENS, WHY THEY
ARE PROCESSED AND PRESENTED.
THE RANDOM FOREST MODEL THAT WE
DEVELOPED BASED ON 147 KNOWN
CLASS I NEOEPITOPES REVEALS
FEATURES THAT ARE ASSOCIATED
WITH NEOANTIGEN RECOGNITION.
SO AS YOU GO TO THE LEFT OF THIS
GRAPH, THOSE ARE THE HIGHEST
AFFINITY PEPTIDES, AND THEN
GOING UP THE Y AXIS, THAT'S THE
MOST HIGHLY EXPRESSED ANTIGENS.
YOU CAN SEE ON THE UPPER
LEFT-HAND CORNER THERE'S AN
ENRICHMENT OF REACTIVITY AGAINST
PEPTIDES, THIS IS CLASS I
PEPTIDES THAT HAVE HIGH
EXPRESSION AND ARE VERY STRONG
BINDERS.
THIS WAS PRESENTED IN THE 2021
NATURE CANCER PAPER FROM OUR
GROUP.
CURRENTLY WE HAVE OVER 700
NEOANTIGENS THAT HAVE BEEN
IDENTIFIED AND OVER 300 CLASS I
NEOEPITOPES SO WE'VE MORE THAN
DOUBLED THE NUMBER OF TARGETS
HERE.
SO WE'RE HOPING TO UPDATE THIS
SOON.
USE SOME OTHER TOOLS, SUCH AS
THE MATCHING ALGORITHM THAT WAS
DEVELOPED BY STERNBERG AND HER
GROUP AND PUBLISHED IN A PAPER
IN IMMUNITY IN 2023.
SO WE'RE GOING TO RE-RUN THE
SYSTEM, RE-RUN THE MODEL AND TRY
TO SEE IF WE CAN IMPROVE OUR
ABILITY TO IDENTIFY THESE
TARGETS BECAUSE I THINK WHEN
YOU'RE TALKING ABOUT MINIMAL PEP
TIE, THERE'S A LARGE UNIVERSE OF
PEPTIDES, ANY ONE MUTATION CAN
GIVE RISE TO OVER 50 -- WE NEED
TO REALLY NARROW DOWN THE
CANDIDATES.
AND WE'RE ACTIVELY EXPLORING THE
USE OF THOSE MINIMAL PEPTIDES AS
METHODS TO IDENTIFY NEW
REACTIVITIES BY IN VITRO
SENSITIZATION OF TIL PERIPHERAL
BLOOD, SEE IF WE CAN INCREASE
THE UNIVERSE OF REACTIVITIES,
WHICH I THINK IS REALLY GOING TO
BE IMPORTANT FOR -- PARTICULARLY
FOR TREATING GI CANCER PATIENTS.
SO WHAT ARE SOME OF THE FUTURE
OPPORTUNITIES?
TO DEVELOP MORE
EFFECTIVE TREATMENTS?
I THINK THE IDENTIFICATION OF
ADDITIONAL TARGETS FOR THERAPY
AS I SAID SEVERAL TIMES IS GOING
TO BE IMPORTANT PARTICULARLY FOR
THESE EPITHELIAL CANCERS.
CERTAINLY REACTIVITY AGAINST
SELF-ANTIGENS CAN CONTRIBUTE AND
ACT USING T CELLS ENGINEERED TO
RECOGNIZE NYESO1 LED TO -- RATE
OF MORE THAN 50% IN PATIENTS
WITH MELANOMA, SYNOVIAL CELL
SARCOMA.
I THINK THE COMBINED PROTEOMIC,
RNA SEQ AND DGE ANALYSIS REALLY
CAN FACILITATE IDENTIFICATION OF
ADDITIONAL TARGET ANTIGENS, BUT
PARTICULARLY WHEN YOU'RE TALKING
ABOUT THE DARK MATTER OUTSIDE OF
THE NORMAL EXOME BECOMES
COMPUTATIONALLY VERY INTENSIVE.
ONLY 2% OF THE GENOME ENCODES
KNOWN EX-ONICS SEQUENCES.
SO YOU'RE TALKING ABOUT THE 98%
OF THE 3 BILLION BASES OF THE
HUMAN GENOME, THE NEED TO BE
EXPLORED.
SO IT'S COMPUTATIONALLY
INTENSIVE, AND WE HAVE A
BIOINFORMATICS GROUP THAT WE
HOPE TO EXPAND TO REALLY ADDRESS
SOME OF THESE QUESTIONS BECAUSE
I THINK THAT'S REALLY GOING TO
BE IMPORTANT AS WE TRY TO
INTEGRATE ALL OF THESE
APPROACHES.
AND OF COURSE FINALLY, WE WANT
TO UNDERSTAND WHAT FACTORS ARE
ASSOCIATED WITH IMMUNE RESPONSE.
SO WHAT ARE THE ANTIGENS WE
SHOULD BE TARGETING, WHAT ARE
THE ONES MEDIATING THESE
EFFECTIVE IMMUNE RESPONSES?
IS IT THE EXPRESSION OR THE
CLOAALITY OF THOSE ANTIGENS
THAT'S KEY, THE ROLE OF THOSE
ANTIGENS OR POTENTIALLY OTHER
FACTORS THAT COULD INFLUENCE THE
ABILITY TO RESPOND.
TO TREATMENT.
AND I'D JUST LIKE TO FINISH BY
ACKNOWLEDGING ALL THE SURGERY
BRANCH RESEARCH STAFF.
THERE'S TOO MANY PEOPLE TO PUT
ON THE SLIDE.
AND THE CLINICAL STAFF,
PATIENTS, AND OF COURSE
DR. ROSENBERG, WITHOUT WHOM THIS
WORK WOULD NOT BE POSSIBLE.
THANK YOU.
[APPLAUSE]
>> QUESTION FOR DR. ROBBINS?
>> HI, PAUL.
-- FROM TORONTO.
OLD FRIEND.
SO I AGREE WITH YOU ABOUT THE
NON-CODING RETRO TRANSPOSABLE
ELEMENTS.
I THINK THAT'S GOING TO BE A
HUGE AREA.
WE'VE BEEN LOOKING AT THAT IN
VARIOUS RARER CANCERS TOO AS
WELL.
THERE'S A HUGE AMOUNT OF
EXPRESSION AND PEPTIDES THAT ARE
PRESENTED, SO I WOULD DEFINITELY
BARK UP THAT TREE.
I HAD SENT A QUESTION, A LOT OF
THE FOCUS HAS BEEN ON CD8 IN
CLASS I, BUT WHAT ABOUT CLASS II
NEOANTIGENS, WHAT INRHODES ARE
BEING MADE?
ARE WE KIND OF BARKING UP THE
WRONG TREE BECAUSE A LOT OF
TUMORS DON'T EXPRESS A LOT OF
CLASS II BUT DENDRITIC CELLS
EXPRESS A LOT AND THEY'RE
NECESSARY FOR HELPING T CELLS.
SO SHOULD WE BE SCREENING MORE
NEOANTIGEN REACTIVE CD4 CELLS
THAT ARE ACTUALLY WHERE THE
PEPTIDES ARE BEING PRESENTED ON
DENDRITIC CELLS AND NOT NOT
NECESSARILY ON THE TUMOR.
>> WELL, WE ARE.
OUR SCREENING APPROACHES USE
LONG PEPTIDES, SO THOSE WE THINK
CAPTURE ESSENTIALLY ALL OF THE
CD4 REACTIVITY.
AND WE FIND IN GI CANCER
ACTUALLY A PREPONDERANCE OF
CD4 REACTIVITY, OR MAYBE IT'S
FAIRLY EQUAL IN TERMS OF THE CD.
SO WE ARE ADMINISTERING T CELLS,
CLASS II RESTRICTED T CELLS AND
IN SOME CASES WE'RE ACTUALLY
ADMINISTERING EXCLUSIVELY, AS
FAR AS WE KNOW, NEOANTIGEN
REACTIVE CD4 CELLS.
AND OF COURSE WE HAVE A
CHOLANGIOCARCINOMA PATIENT WHO
FAMOUSLY RESPONDED TO A CD4
REACTIVE T-CELL.
SO WE CERTAINLY DON'T WANT TO
DISCOUNT CD4 CELLS.
WE'RE ALSO EXPLORING THE USE OF
CLASS II RESTRICTED T-CELL
RECEPTORS.
WHETHER THOSE ALONE WILL BE
EFFECTIVE, WE DON'T KNOW.
WE'VE ONLY TREATED A FEW
PATIENTS SO IT'S STILL TOO
EARLY.
BUT PERHAPS THE COMBINATION OF
THE TWO WILL BE IMPORTANT.
WHICH AGAIN, WE JUST NEED TO
FIND MORE TARGETS BECAUSE YOU
NEED TO HAVE A PATIENT WHERE YOU
CAN MATCH BOTH.
AS I POINTED OUT WE HAVE KRAS
AND P53 CLASS I AND CLASS II
RESTRICTED T CELLS BUT THOSE ARE
NOT NECESSARILY IDENTIFIED IN
THE SAME PATIENT BECAUSE YOU
HAVE TO HAVE THE RIGHT
RESTRICTION ELEMENTS.
>> I GUESS WHAT I WAS SAYING IS
LIKE NEOANTIGEN CLASS II
NEOANTIGEN HELPER EPITOPES AS
OPPOSED TO DIRECT -- ANYWAY, WE
CAN TALK ABOUT IT LATER.
>> OKAY, WE CAN TALK ABOUT IT
LATER.
>> QUICK QUESTION.
SO PAUL, GREAT TALK.
THOSE T-CELL RECEPTORS YOU FIND
THAT ARE NOT NEOANTIGEN
RESTRICTIVE, HAVE YOU LOOKED AT
THOSE TO SEE HOW BROADLY THEY
REACT TO OTHER HLA MATCHED
TUMORS?
>> WE'RE JUST STARTING THOSE
STUDIES.
SO I THINK WE DO IN A FEW CASES
HAVE EVIDENCE THAT THEY CAN
RECOGNIZE MORE THAN ONE, BUT WE
DON'T HAVE ENOUGH DATA TO SAY.
>> SO WE'LL PRESENT A LITTLE BIT
OF DATA AT CTSI THAT APPEARS TO
BE HLA RESTRICTED IN A
B44 SUPERTYPE SO THERE ARE
THINGS THERE THAT ARE GOING TO
BE REALLY FASCINATING.
I THINK THEY'RE RELATED BACK TO
DARK MATTER.
>> SO WE'RE GOING TO MOVE ON
WITH OUR NEXT SPEAKER, DR. JIM
YANG, WHO'S BEEN A MEMBER OF THE
SENIOR STAFF HERE FOR 40 YEARS.
SO I BELIEVE LIKE DR. ROSENBERG,
JIM, THIS HAS BEEN YOUR FIRST
AND ONLY JOB?
>> YES, FIRST AND ONLY JOB.
[LAUGHTER]
I HOPE IT'S MY LAST.
I'D LIKE TO START OUT JUST BY
THANKING STEVE.
STEVE HAS CREATED A SCIENTIFIC
PARADISE HERE, AND HE HAS
ASSEMBLED A GROUP OF DEDICATED,
INTELLIGENT REALLY GOOD HEARTED
PEOPLE WHO UNDERSTAND THE
MESSAGE, AND MOST OF THEM ARE IN
THIS ROOM.
IT'S REALLY QUITE REMARKABLE,
AND HE SHARED WITH THEM THIS
UNIQUE EXPERIENCE OF USING YOUR
SKILLS TO TREAT A PATIENT WITH
INCURABLE CANCER AND RETURN THEM
TO A COMPLETELY NORMAL LIFE.
ONCE YOU'VE SEEN THAT, YOU NEVER
WANT TO DO ANYTHING ELSE.
SO THE NIH FACILITIES AND
STEVE'S LEADERSHIP HAS LED TO
THIS HAPPENING AGAIN AND AGAIN,
AND IT'S MADE MY LIFE A BLESSED
LIFE.
IT REALLY HAS.
SO I'D LIKE TO THANK YOU, STEVE.
SO I'M GOING TO BE TALKING ABOUT
TARGETING NEOANTIGEN WITH T
CELLS.
I'M GOING TO START OUT WITH,
LIKE JIM MULE,, THE PRETTIER THE
SLIDE, THE LESS COMPELLING THE
EVIDENCE.
SO WE ALWAYS JUST DID BLACK AND
WHITE SLIDES.
UNFORTUNATELY THERE ARE SOME
COLOR BACKGROUNDS THAT HAVE
SLIPPED IN, I'LL HAVE TO WORK ON
THAT.
IT'S TITLED STOCKING THE TCR
LIBRARY AGAINST DRIVER
MUTATIONS.
SO THE IDEA IS DISARMINGLY
SIMPLE OF ADOPTIVE T-CELL
TRANSFER, IS TO ADMINISTER TUMOR
REACTIVE T CELLS EXPANDED IN
VITRO TO CAUSE THE IMMUNOLOGIC
REJECTION OF METASTATIC CANCERS.
YOU CAN DO THIS WITH ORGANS,
SKIN GRAFTS AND SO FORTH, SO THE
QUESTION IS WHY NOT CANCER.
NOW THE THEORETICAL ADVANTAGES
FOR WHICH PROBABLY WE'RE THE
BEST KNOWN IS THAT YOU CAN
SELECT THE CLONE TYPES YOU CAN
ADMINISTER, YOU CAN USE REAGENTS
THAT ARE NOT TOLERATED IN VIVO,
CAN YOU ALSO PREPARE THE
RECIPIENT AND THE T-CELL INKNOCK
LANTS SEPARATELY BECAUSE THEY
OFTEN HAVE VERY CONFLICTING
NEEDS, AND YOU CAN MODIFY THE T
CELLS ADMINISTERED, SOMETHING
THAT STILL NEEDS TO BE EXPLORED
FURTHER THAT I'LL MENTION A
LITTLE BIT AT THE END.
SO YOU'VE HEARD FROM YUTAKA AND
PAUL ABOUT THE UNMUTATED SELF
ANTIGENS CHARACTER USED BY THE
MELANOCYTIC DIFFERENTIATION
PROTEINS AND YOU'VE ALSO HEARD
FROM PAUL DID -- IF YOU LOOK AT
THOSE TWO CATEGORIES WHAT YOU
SEE IS THAT THIS IS CONSTANT
BETWEEN PATIENTS.
THAT'S ITS REAL ATTRACTION.
PEOPLE LIKE IT BECAUSE YOU CAN
MAKE OFF THE SHELF REAGENTS.
BUT THE DOWNSIDE IS THERE'S A
POTENTIAL FOR AUTOIMMUNE
TOXICITY BECAUSE THESE ARE
NORMAL SELF PROTEINS AND THE
T-CELL REPERTOIRE CAN BE LIMITED
BY THYMIC EDITING.
NOW THE MUTATED NON-SELF
NEOANTIGENS ARE HIGHLY
PATIENT-SPECIFIC.
SO THAT'S VERY COMPLICATED.
BUT THEY HAVE VERY LOW POTENTIAL
FOR AUTOIMMUNITY, NO CENTRAL
THYMIC TOLERANCE, AND MOST
TUMORS AS PARKER AND OTHERS HAVE
SHOWN HAVE IMMUNOGENIC
NEOANTIGENS.
SO WHAT YOU'D REALLY LIKE IS
BOTH.
WHAT YOU'D LIKE TO TARGET THESE
UNIQUE NEOANTIGENS BUT ONES THAT
END UP BEING SHARED BETWEEN
DIFFERENT HUMAN CANCER, SO
THAT'S WHAT I'M GOING TO BE
TALKING ABOUT TODAY, IS
TARGETING RECURRENT ONCOGENIC
MUTATION.
WE KNOW THIS FROM THE PAPER
LOOKING AT A PATIENT TREATED
WITH TIL WHICH HAD REACTIVITY
AGAINST THE KRAS G12D MUTATION
PRESENTED BY H LACO802 AND SHE
HAD THIS REALLY GOOD RESPONSE OF
MULTIPLE LUNG METASTASES.
THIS WENT AWAY, THIS ONE WENT
AWAY, THIS ONE SHRUNK.
BUT THIS ONE IN THE THIRD ROW
GREW INITIALLY AND THEN --
SHRANK INITIALLY AND THEN GREW
BACK.
SO THIS SHOWS, THOUGH, THAT YOU
COULD INDEED TARGET THESE
EFFECTIVELY IN THIS CASE WITH
TIL.
SO WHAT I'M GOING TO BE TALKING
ABOUT IS GENERATING TCRs THAT
ARE OFF THE SHELF REAGENTS
AGAINST DRIVER MUTATIONS AND WE
USE HLA TRANSGENIC MICE FOR
THIS.
THE FIRST PAPER WE PUBLISHED WAS
AT THE SAME TIME THE CLINICAL
TIL PATIENT WAS PUBLISHED IN
2016.
THEY LOOKED AT IDENTIFYING THEM
USING TRANSGENIC MICE.
TRANSGENIC MICE HAVE HUMAN HLA
AND WE HAVE AT LEAST THREE
STRAINS THAT EXPRESS HLA0201,
A1101 AND WE MADE THE HL
HLA0301 MOUSE STRAIN.
THEY'RE IMMUNIZED AND YOU
HARVEST THEIR SPLEENS AND
DRAINING LYMPH NODES AND
STIMULATE THEM INVIE TROA.
IN VITRO.
YOU CAN ISOLATE THEM AND THEN
YOU CLONE THE T-CELL RECEPTORS
AND REEXPRESS THEM IN HUMAN PBL
AND TEST THEM.
WHEN SHE DID THIS FOR HER FIRST
TWO KRAS SPECIFICALLY AL11
RESTRICTED TCRs, SHE FOUND
THAT THEY DID WORK IN MOUSE
MODELS, WHERE THIS IS A MOUSE
MODEL WHERE A TUMOR IS
EXPRESSING THE G12V MUTATION IN
HLA-A11 TRANSFECTED TUMOR SO
THAT WHAT YOU SEE IS THAT
STARTING WITH PALPABLE
MACROSCOPIC TUMORS THAT YOU
COULD GIVE THESE CELLS AND CAUSE
REGRESSION ON THE BOTTOM LINE
THAT DID NOT OCCUR WITH NO
TREATMENT UNTRANSDUCED CELLS OR
MOCK TRANSDUCED CELLS.
SO THEY WERE EFFECTIVE IN THE
XENOGRAPH MODELS.
NOW WHY ARE WE LOOKING AT MOUSE
RECEPTORS?
WE KNOW FROM OUR OWN RECEPTORS
THAT THEY WORK IN HUMAN PBLs.
YOU CAN PUT THEM IN WITH NO
MODIFICATION, GIVE TO THAT
PATIENT AND THEY WILL WORK.
THESE ARE THREE SETS OF
PROTOCOLS WE PUBLISHED USING
MURINE RECEPTORS AGAINST
DIFFERENTIATION ANTIGENS,
AGAINST NORMAL -- IN ALL OF
THESE WE DID SEE -- WE ALSO SAW
AUTOIMMUAUTOIMMUNE TOXICITIES.
THIS IS THE DANGER OF COURSE OF
TARGETING NON-MUTATED SELF
ANTIGENS.
SO OUR APPROACH WAS TO LOOK AT
MUTATED NEOANTIGENS THAT WERE
SHARED.
SO THIS IS A STUDY THAT WAS DONE
BY CATHERINE AEDE LOOKING AT
SYSTEMATIC IDENTIFICATION BY
MASS SPEC.
THE APPROACH WAS TO TRANSFECT
CELL LINES WITH MUTATED MINI
GENES EXPRESSING THE TARGET AND
THE ALLELE OF INTEREST.
THESE WERE MHC NULL LIELS TO
SIMPLIFY THE ELUSION OF
PEPTIDES.
WE USE LINES WITH BOTH THE
STANDARD PROTEOSOME AND THE
IMMUNOPROTEOSOME AND TYPICALLY
DIDN'T FIND MUCH DIFFERENCE BUT
OCCASIONALLY SEE IT, AND WE THEN
IMMUNOPRECIPITATE THE HLA AND EE
LUTE THE PEPTIDES AND ANALYZE
THEM BY MASS SPEC.
WE PICK THESE RECURRING
MUTATIONS BECAUSE THEY'RE THE
MOST COMMON ONES IN HUMAN
CANCERS WITH VERY SPECIFIC
RECURRING AMINO ACID
SUBSTITUTIONS.
IN RAS, EGFR -- YOU CAN SEE THE
VARIOUS MUTATIONS LISTED HERE
AND WHICH CANCERS THEY OCCUR IN.
WE THEN PICK THE DOZEN MOST
COMMON CLASS I ALLELES IN THE
U.S. POPULATION.
AND WANTED TO INVESTIGATE THOSE.
BUT THAT RESULTS IN HUNDREDS OF
COMBINATIONS OF MUTATIONS AND
ALLELES SO WE NEEDED A METHOD TO
FIGURE OUT WHICH ONES WERE
WORTHY OF INVESTIGATIONS AND
WHICH ONES WERE NOT.
SO THIS IS AN EXAMPLE OF HOW IT
LOOKED.
THIS IS -- ACROSS THE TOP IS
LOOKING AT THE E545K MUTATION IN
PIC3CA.
WE PUT IT INTO THE TWO LINES,
THE TOP ONE BEING THE
IMMUNOPROTEOSOME LINE, THE
BOTTOM BEING THE STANDARD
PROTEOSOME LINE.
THE PEPTIDE, MUTATED PEPTIDE IS
SHOWN IN THE THIRD COLUMN, AND
THEN THE GREEN BOX TO THE RIGHT
REPRESENT INDIVIDUAL REPLICATES,
BIOLOGICAL REPLICATES OF THESE
EXPERIMENTS.
SO CATHERINE DID EVERY ONE OF
THESE UP TO EIGHT TIMES, AND
WHAT YOU SEE, IF IT'S DARK
GREEP, I GREEN, IT MEANS THE
MUTATED PEPTIDE WAS RECOVERED
ABUNDANTLY AND CONSISTENTLY.
LIGHT GREEN MEANS IT WAS A
LITTLE LESS CONSISTENT OR
ABUNDANT.
RED MEANS NO CANDIDATE MUTATED
PEPTIDES WERE RECOVERED AND
WHITE MEANS IT HASN'T BEEN DONE
YET.
AND WE DID ALSO FRACTIONATION
THAT SHOWS BELOW THE PEPTIDE WE
EE LUTE AND IDENTIFY IS INDEED
THE PEPTIDE WE THINK IT IS.
SO THIS IS A SUMMARY OF THE HLA
A11 PEPTIDES FROM FOUR OF THE
COMMON MUTATED DRIVER GENES.
YOU CAN SEE MANY OF THEM ARE
WELL PROCESSED AND PRESENTED BY
BOTH OF THE LINES SHOWN HERE.
OCCASIONALLY YOU'LL SEE SOME
THAT ARE NOT.
THE BRAF -- WAS POORLY PRESENTED
BUT THE OTHERS WERE WELL
PRESENTED.
HLA A3 ALSO VERY GOOD PRESENTER
OF MANY OF THESE AND
OCCASIONALLY SOME LIKE YOUR G12D
PRESENTED BY A3 WAS POORLY
PRESENTED AND AGAIN, ONE WE DID
NOT PURSUE.
SO THIS ALLOWED US TO FOCUS ON
WHICH ONES WERE WORTHY OF
EFFORT.
THEY WEREN'T ALL POSITIVE.
HLA A11 DID NOT PRESENT THESE
TWO EPITOPES AND HLA A3 DIDN'T
PRESENT THE THREE BELOW.
THE OTHER THING THAT WAS
INTERESTING WAS THE A2 PRESENTED
EPITOPES WERE NOT VERY GOOD.
WE RARELY FOUND THESE PRESENTED
BY A2 SO WE FOCUSED MIME PRIMARN
A3 AND -- 11.
YOU DO THE VACCINATION, YOU
HARVEST THE SPLEENS AND LYMPH
NODES OF THESE HLA TRANSGENIC
MICE AND THEN YOU TRY AND SELECT
THE REACTIVE CELLS AND WE USED A
VARIETY OF TECHNIQUES, BUT ONE
THAT WAS VERY HELPFUL WAS USING
TETRAMERS.
WHAT I SHOW HERE IS THE TWO
DIFFERENT COLOR TETRAMERS ON THE
X AND Y AXIS.
THIS IS UNSTAINED.
OVER HERE IS THE TETRAMER
STAINING.
YOU CAN SEE THERE'S DOUBLE
POSITIVE TETRAMER EXPRESSING
CELLS.
YOU SORT FOR THESE, YOU THEN
CLONE THEIR T-CELL RECEPTORS AND
YOU GET THESE T-CELL RECEPTOR
ALPHA BETA PAIRS OVER HERE, YOU
RECONSTRUCT THEM IN PBL AND THEN
YOU TEST THEM.
WHEN YOU DO THAT, YOU SEE HERE
THE MUTANT PEPTIDE TITRATIONS OF
ONE, THIS IS THE HLA A11
RESTRICTED EGFRL858R MUTATION
AND WHEN YOU DO THAT, YOU CAN
SEE THAT THESE THREE RECEPTORS
WE ISOLATED WERE ALL SPECIFIC
FOR THE MUTANT PEPTIDE WITH
LITTLE TO NO RECOGNITION OF WILD
TYPE.
AND WHEN YOU DO THIS IN A
TRANSFECTED K562A11 EXPRESSING
CELL, YOU CAN SEE THAT PUTTING
IN THE TARGET AND ALLOWING TO BE
NATURALLY PROCESSED ALSO RESULTS
IN RECOGNITION WITH NO
RECOGNITION OF JUST HLA POS HIF
TARGETS WITHOUT THE ANTIGEN.
NOW, WHEN WE DID THAT, THIS IS
ANOTHER RECEPTOR, AGAIN TOTALLY
SPECIFIC FOR A MUTATED EPITOPE,
NO WILD TYPE REACTIVITY.
BUT WHEN WE TEST IT A PANEL OF
TUMORS THAT ARE NATURALLY
EXPRESSING THIS, THESE THREE
TUMORS EXPRESS HLA AND THE G12B
KRAS MUTATION.
ALL THREE SHOW SOME REACTIVITY.
THE TWO NEGATIVE CONTROLS FOR
HLA AND THE ANTIGEN ARE
NEGATIVE, BUT YOU CAN SEE THESE
ARE NOT VERY POTENT.
THIS AMOUNT OF INTERFERON GAMMA
ELABORATION IS VERY LOW FOR OUR
TYPICAL TUMOR REACTIVE T-CELL.
THE SAME WAS THROUGH FOR ANOTHER
ONE, A3 PRESENTED EPITOPE.
AGAIN WE HAD THREE TCRs, ALL
OF THEM WERE HIGHLY RECOGNIZING
THE MUTANT EPITOPE, BUT TWO OF
THEM HAD SOME WILD TYPE
REACTIVITY, TCR 4 HAD NONE.
WHEN YOU TAKE TCR 4 AND TEST IT
AGAINST THIS PANEL OF TARGETS,
AGAIN, THE SPECIFIC CORRECT
TUMORS ALL RELEASE SOME
INTERFERON, THE TWO NEGATIVE
CONTROLS DO NOT.
BUT AGAIN, VERY LOW LEVELS OF
INTERFERON.
AND SO AND THIS WAS TRUE FOR
MOST OF OUR TCRs WE FOUND.
THIS IS THE SET OF FOUR OF THEM
FOR HLA A11 AND I LIST THESE
FOUR BECAUSE WE HAVE NO HUMAN
HOMOLOGUE FOR THESE.
WE'VE NEVER FOUND THESE
REACTIVITIES IN HUMAN TIL.
SO WHEN WE WANT TO DO A T-CELL
RECEPTOR PROTOCOL, WE'VE BEEN
USING HUMAN TCRs BUT WE DO NOT
HAVE THEIR EQUIVALENT FOR THESE
IN HUMAN TCRs.
SO AGAIN, YOU CAN SEE THEY'RE
EXTREMELY ECK QIS ITALY SPECIFIC
FOR THE MUTANT ALLELE BUT NOT
FOR THE WILD TYPE.
THIS IS THE A3.
AGAIN, ALL OF THESE WERE VERY
SPECIFIC EXCEPT FOR THE ONE I
SHOWED YOU AGAINST BRAF -- TWO
OF THEM HAD WILD TYPE REACTIVITY
BUT RECEPTOR 4 DID NOT.
SO THE QUESTION IS CAN WE DO
SOMETHING ABOUT THIS.
CAN WE ENHANCE THE REACTIVITY OF
THESE TCRs TO FILL THESE
VACANCIES IN OUR DRIVER
NEOANTIGEN TCR LIBRARY.
SO WE TRIED MODIFYING CDR3s AS
PAUL HAS DONE PREVIOUSLY
SUCCESSFULLY.
WE ADDED CHIMERIC CORECEPTORS TO
THE TCRs.
WE MODIFIED THE SIGNALING IN
THESE T CELLS AND NONE OF THESE
INTERVENTIONS HAD CONSISTENT OR
SIGNIFICANT BENEFIT.
SO FINALLY WE TURNED TO THE IDEA
OF ADDING IL12 TO THE TUMOR
MICROENVIRONMENT.
WE DID THIS BECAUSE WE HAD A
PREVIOUS EXPERIENCE IN THE
BRANCH USING AN N FAD DRIVEN
PROMOTER TO CREATE A SINGLE
CHAIN IL2 MOLECULE.
THIS IS WITH LING SANG AND RICK
MORGAN AND THEY REPORTED THAT
THERE ARE 33 PATIENTS WITH MEL
MELANOMA ENROLLED.
ONE PERSON HAD A TRIVIAL CELL
DOAN'S, 30 MILLION TIL, HAD A
COMPLETE DURABLE AND COMPLETE
RESPONSE AND THE OVERALL
RESPONSE RATE WAS 6 63% WITH AG
LESS CELLS IN MANY CASES THAN WE
TYPICALLY GIVE.
SO THIS WAS VERY ENCOURAGING
INITIALLY BUT AT HIGHER DOSES
PATIENTS EXPERIENCED CYTOKINE
STORM AND HEPATOTOXICITY THAT
LIMITED OUR ABILITY TO GIVE
THOSE CELLS.
THIS IS THE PATIENT I MENTIONED,
QUITE A REMARKABLE PATIENT
BECAUSE SHE HAD BEEN TREATED
WITH CONVENTIONAL TIL AND IL-2.
SHE GOT 30 BILLION CELLS WITH
SEVERAL DOSES OF IL-2, YOU CAN
SEE HER TUMOR GREW RIGHT THROUGH
THAT.
WE THEN USED THE EXACT SAME TIL
USING THE -- IL12 AND SHE ONLY
GOT -- SHE HAD THIS VERY
DRAMATIC RESPONSE.
SO WE WERE VERY ENCOURAGED BY
THAT BUT WE WERE WORRIED ABOUT
THE SAFETY.
SO WE'RE LOOKING AT TRYING TO
ADD IL-2 TO CELLS SAFELY, AND
INITIALLY LYNN ZANG SHOWED THERE
WAS INCREASED SAFETY IF YOU
MEMBRANE TETHERED THE -- MICE
DID NOT GET SICK WITH THAT.
PETER THEN DEVELOPED AN NFAT
DRIVEN MEMBRANE TETHERED IL-12
INCORPORATING A DEGRON DOMAIN,
SO THIS IS YET AN ADDITIONAL
SAFETY MEASURE THAT YOU CAN USE
IN THIS CASE.
AND WE TRIED THIS THEN IN OUR
MURINE TIL PBL.
SO IL-12 CAN ENHANCE THE IN
VITRO FUNCTION OF TUMOR-REACTIVE
CELLS BOTH IN MICE AND
APPARENTLY IN HUMANS AS WELL.
THE DOMINANT KNOWN MECHANISM IS
THROUGH INTERFERON GAMMA.
IT AUGMENTS INTERFERON GAMMA, IT
BIASES CELLS TO A TH1
TC1 PHENOTYPE BUT LITTLE IS
KNOWN BEYOND THAT WHAT IT DOES
TO OTHER FUNCTIONS.
-- OR ONE THAT IS REACTING
AGAINST A VERY SPARSE ANTIGEN.
IT'S ALSO NOT CLEAR WHAT IT
WOULD DO TO OTHER EFFECTOR
FUNCTIONS SUCH AS CYTOTOXICITY.
SO WE ADDED IT, CATHERINE AND
PETER TOGETHER ADDED THIS
CONSTRUCT TO THE PBL EXPRESSING.
IN THIS CASE THE MURINE TIL
AGAINST THE G TELL VA3 REA
STRICTED RECEPTOR FOR WHICH WE
DO NOT HAVE A HUMAN EQUIVALENT.
WHAT YOU SEE HERE IS THAT THIS
IS THE RECEPTOR BY ITSELF.
AGAIN, A FEW HUNDRED DOLLARS
PICOGRAMS OF INTERFERON GAMMA
ELABORATED AGAINST THE MUTANT
ALLELE, BUT WHEN YOU ADD THE
DEGRON DOMAIN AGAINST THIS
TUMOR, YOU GET A HUGE
AUGMENTATION OF THE INTERFERON
GAMMA SECRETION.
AND THIS IS TRUE FOR, AGAIN,
THIS TUMOR LINE AGAIN POSITIVE
FOR THE ANTIGEN AND THE MHC AND
THIS TUMOR LINE INTO THE
THOUSANDS INSTEAD OF HUNDREDS OF
PICOGRAMS OF INTERFERON.
AND WITHOUT LOSS OF SPECIFICITY.
THESE ARE THE SPECIFICITY
RESULTS FOR THE HLA AND THE
ANTIGEN.
SO THIS WAS A ENCOURAGING
AUGMENTATION WITHOUT LOSS OF
SPECIFICITY.
IF YOU THEN ADD THIS ALSO TO A
DIFFERENT RECEPTOR, THE TCR
4V600E RECEPTOR, I SHOWED YOU
THE SAME THING, AGAIN, THIS IS
VERY LOW REACTIVITY WE SAW
INITIALLY, THIS IS THE HIGHLY
AUGMENTED SECRETION OF
INTERFERON GAMMA, AND THE
SPECIFICITY REMAINED VERY GOOD
AGAINST THE WILD TYPE ALLELE.
SO THE QUESTION -- SO THIS IS
ENCOURAGING.
THE QUESTION IS WHAT DOES DO TO
OTHER EFFECTOR FUNCTIONS? IF
YOU LOOK AT CYTOTOXICITY, THIS
IS THE FIRST KRAS REACTIVE
RECEPTOR, THIS IS THE DRK I'M
SORRY BRAF AND THIS IS THE K
RAF.
THE TCR ALONE SHOWS THIS BLUE
LINE REACTIVITY AND THE TUMOR
CELLS GROW OUT PROGRESSIVELY
WITH VERY LITTLE DIFFERENCE
BETWEEN THE CONTROLS SHOWING THE
AMOUNT OF KILLING HERE.
BUT WHEN YOU PUT IN THE IL12
DOMAIN, IT ACTUALLY TURNS INTO
AN EXCELLENT KILLER.
SAY TOE TOXICITY COMPARABLE TO
OTHER RECEPTORS THAT ARE
SUCCESSFUL.
AGAINST THE KRAS EPITOPE, AGAIN,
THERE WAS REALLY NO CYTOTOXICITY
OF THE RECEPTOR BY ITSELF, BUT
WITH THE DEGRON DOMAIN, THERE
CLEARLY IS CYTOTOXICITY.
IT WASN'T QUITE AS GOOD AS THIS
AUTOLOGOUS EFFECTOR CLONE FROM
THE SAME PATIENT AND FROM THE
PATIENT WHO PRODUCED THIS CELL
LINE, BUT IT CLEARLY IS
CYTOTOXICITY THAT WASN'T PRESENT
BEFORE.
SO THIS IS VERY ENCOURAGING THAT
IT IS AUGMENTING OTHER FUNCTIONS
BESIDES INTERFERON GAMMA.
SOME PEOPLE ASK WHY ARE YOU
TRYING TO RESCUE WEAK TCRs?
THE ANSWER TO THAT IS STRONG
ANTITUMOR RESPONSES CAN GENERATE
PRIMARY RESISTANCE THROUGH TUMOR
EDITINGMENT SOME OF THESE ESCAPE
MECHANISMS SUCH AS ANTIPD-1 ARE
REVERSIBLE.
IF A T-CELL EXPRESSES PD-1, IT
CAN BE BLOCKED BY ANTIPD-1, BUT
OTHERS ARE NOT.
YOU'VE HEARD A LOT IN THE
CLASS I SYSTEM FOR LOSS OF THE
PRESENTING ALLELE FOR THE TUMOR
ANTIGEN.
AND FOR CLASS I CELL THERAPY,
THAT'S THE KISS OF DEATH, THAT
NO LONGER WORKS.
SO THAT LED ME TO MY FINAL POINT
WHICH IS TALKING ABOUT THIS
SURPRISE ATTACK HYPOTHESIS.
THIS ORIGINALLY CAME UP BECAUSE
I WONDERED WHY DO CAR-T CELLS
AGAINST B CELL AND PLASMA CELL
MALIGNANCIES WORK SO WELL WITH
ONLY A FEW MILLION CELLS?
THAT'S A TYPICAL WHOLE BODY DOSE
WE GIVE TO PATIENT WITH THOSE
CARS.
THAT'S A THOUSAND FOLD FEWER WE
GIVE WITH TILL USING NORMAL
T-CELL RECEPTORS AGAINST A SOLID
TUMOR ANTIGENS.
AND THE ANSWER I ALWAYS GET WAS,
WELL, THE CARs ARE TREATING
LIQUID TUMORS, MORE ACCESSIBLE
TO T CELLS, OR THAT THE TARGET
TUMORS ARE HEMATOPOIETIC CANCERS
AND THEY STIMULATE OR
CO-STIMULATE T CELLS.
SO LOOKING AT THAT, THESE ARE --
THIS IS THE PARADOX.
THIS IS THE TIL AT THE TOP WHERE
WE GIVE 30 TO 150 BILLION CELLS,
WE GET VERY GOOD RESPONSE RATES
IN MELANOMA.
IF YOU LOOK AT THE TWO CAR CD19
AND BCMA, YOU'RE GIVING TWO TO
THREE LOGS LESS CELLS TO GET
THOSE RESPONSE RATES.
AND THE ANSWER WAS ALWAYS, OH,
JUST A LIQUID TUMOR.
BUT THEN THERE ARE NOW A COUPLE
OF CAR THERAPIES AGAINST SOLID
TUMORS THAT ARE STARTING TO SHOW
ACTIVITY.
NOTICE THE CELL DOSES ARE AGAIN
MICROSCOPIC, 2 TO 3 LOGS LESS,
AND COMPARABLE TO THE CARS FOR
LIQUID TUMORS.
I REMIND YOU ALSO THAT WE HAD A
T-CELL RECEPTOR AGAINST ANOTHER
ANTIGEN, CEA, AGAIN SOLID TUMOR,
CLASSIC T-CELL RECEPTOR, AND
AGAIN, THE DOSE THAT LED TO THIS
TUMOR REGRESSION, WE ONLY
TREATED THREE PATIENTS.
THIS TUMOR REGRESSION, AN AND
FEROCIOUS COLITIS, THAT WAS ALSO
WITH VERY, VERY SMALL NUMBERS OF
CELLS.
THEN FINALLY ARE THE THREE
NEWEST PROTOCOLS LOOKING AT
THESE OTHER SELF-ANTIGENS IN
SOLID TUMORS.
THESE ARE MIXED.
THESE ARE ANTIGENS THAT HAVE
SOME ENDOGENOUS IMMUNE RESPONSES
BUT OFTEN VERY WEAK.
IF YOU CLONE THEM AND AUGMENT
THEIR AFFINITY AND GIVE THEM,
AGAIN, IN SOME CASES, VERY LOW
NUMBERS OF CELLS GIVEN CAN
RESULT IN GOOD CLINICAL
RESPONSES.
SO THE ATTACK -- AND IF YOU'LL
NOTICE, THESE ALL REQUIRED
AFFINITY ENHANCEMENT HERE TO
CAUSE CLINICAL RESPONSES.
SO THE ATTACK HYPOTHESIS.
I SAID STRONG TUMOR NEOANTIGENS
WILL GENERATE PRIMARY
RESISTANCE, NONE OF WIS IS
REVERSIBLE.
THIS STUDY SHOWS IF YOU LOOK AT
ALL KRAS MUTATED PATIENTS IN OUR
DATABASE, YOU INCLUDE HLA A2 --
IT THE MOST COMMON HLA CLASS I
ALLELE AND THESE ARE FOUR
PRESENTING ALLELES THAT DO
PRESENT KRAS MUTATED EPITOPES.
IF YOU LACK AT THE G12D ONE,
THIS IS A NUMBER OF TUMORS IN
RED THAT HAVE LOST THEIR
PRESENTING ALLELE.
YOU CAN SEE THAT IN SOME CASES,
NONE OF THEM LOST IT.
BUT IN OTHER CASES, THERE IS
GROWING LOSS.
THIS IS THE G12V EPITOPE.
YOU SHOULD KNOW THAT THAT IS THE
MOST IMMUNOGENIC KRAS EPITOPE.
WHEN WE WERE VACCINATING OUR
MICE, THAT WAS OUR POSITIVE
CONTROL FOR VACCINATION, BECAUSE
WE ALWAYS GENERATED A GOOD G12V
ANTIEPITOPE RESPONSE.
SO IF YOU LOOK AT THAT, THE MOST
POTENT PRESENTER WITH THE MOST
ABUNDANT PEPTIDE PRESENTATION
WAS THE A11 EPITOPE.
YOU CAN SEE HERE THAT A11 HERE,
HALF OF THE PATIENTS THAT HAVE
THIS COMBINATION OF THIS HIGHLY
IMMUNOGENIC EPITOPE AND THE
MOST -- AND THE BEST PRESENTING
ALLELE HAVE CLASS I LOSS OF A11.
I ALSO SHOW YOU AT THE TOP WITH
THE ASTERISK, THESE ARE THE
EPITOPES AND HLA COMBINATIONS
FOR WHICH WE HAVE HUMAN TCRs
BECAUSE WE FOUND THAT REACTIVITY
IN TIL.
YOU CAN SEE THOSE ARE THE FOUR
WITH THE GREATEST CLASS I LOSS.
SO THIS IS A CLASS I SELECTION
PROBLEM THAT IS NOT REVERSIBLE.
SO THE ATTACK HYPOTHESIS STARTS
WITH THAT, AND THAT CARS AND
SOME TCRs AGAINST NORMAL SELF
ANTIGENS HAVE BEEN EFFECTIVE AT
LOW CELL DOSES BECAUSE THOSE
ATTACK TARGETS HAVE NEVER
ENCOUNTERED IMMUNE ATTACK
BEFORE.
THE P CELLS HA HAVE NOT BEEN
ATTACKED BECAUSE OF CD4 EVER IN
THEIR LIFE AND WEAK -- HAVE THIS
ADVANTAGE AS WELL BUT THEY CAN
BE MADE CLINICALLY EFFECTIVE
WITH PCR ENHANCEMENT.
ALBEIT WITH POTENTIAL HAZARD.
SO -- TO TRY AND ENHANCE THEIR
REACTIVITY BY -- DISPLAY,
PROTEIN PREDICTION STRUCTURAL
STUDIES, THEN TO PRODUCE THIS
ARRAY WITH THE MEMBRANE IL12
DEGRON CONSTRUCT AND IF THEY
SHOW ACTIVITY IN XENOGRAPH
MODELS AND DO NOT SHOW OFF
TARGET CROSS REACTIVITY, THEN WE
WOULD PROCEED TO CLINICAL TRIALS
WITH THOSE.
AND THESE ARE THE PEOPLE
INVOLVED.
THANK YOU.
[APPLAUSE]
>> THANKS, JIM.
OUR NEXT SPEAKER IS DR. ALENA
GROS FROM THE VAIL D'HEBON
INSTITUTE OF ONCOLOGY IN
BARCELONA, ALSO FORMER FELLOW.
>> GOOD MORNING, EVERYONE.
I'M THRILLED TO BE HERE.
THERE ARE TWO WORDS THAT REMIND
ME OF STEVEN ROSENBERG AND
AUTOMATICALLY MAKE ME SMILE.
LASER FOCUS.
HE'S REALLY, REALLY ABLE TO
FOCUS ON HIS SCIENTIFIC GOALS
WITH AN INTENSITY UNLIKE
ANYTHING I'VE SEEN, AND I THINK
IT'S PROBABLY ONE OF HIS SECRET
WEAPONS TO HIS SUCCESSFUL
CAREER.
I REMEMBER MEETING WITH HIM AND
SHOWING HIM SOME DATA AND HIM
SMILING AT ME WITH HIS BLUE EYES
AND SAYING, ALENA, I NEED TO
LASER-FOCUS ON THIS.
NOW MANY YEARS AFTER THAT, I
TELL THIS TO MYSELF PRETTY
FROACTLY, I MUST FREQUENTLY, I .
I'M THRILLED TO BE HERE AND I
CAN'T IMAGINE A BETTER WAY OF
CELEBRATING YOUR CAREER THAN
SHOWING YOU SOME DATA GENERATED
IN MY LAB, SO I'LL BE TALKING
ABOUT THE POSSIBILITY OF EXPL
EXPLORING THE IMMUNOGENICITY OF
NOVEL SOURCES OR TUMOR ANTIGENS.
UNDERSTANDING WHAT TUMOR
ANTIGENS ARE LISTED ANTITUMOR
T-CELL RESPONSES IS CENTRAL TO
CANCER I KNEW KNOW THERAPY
BECAUSE MAINLY UNDERSTANDING
WHICH WILL ENABLE AND HAS
ENABLED THE DEVELOPMENT OF MORE
SAFE AND EFFECTIVE T-CELL
THERAPIES ON VACCINES BUT ALSO
TO UNDERSTAND AND PERSONALIZE
IMMUNE CHECKPOINT INHIBITOR
THERAPIES AND OTHER
IMMUNOTHERAPIES.
CERTAINLY NEOANTIGENS HAVE
EMERGED AS POTENT TARGETS FOR
CANCER IMMUNOTHERAPY, AND WE'VE
HEARD FROM PAUL ROBBINS SOME OF
THE BENEFITS OF TARGETING THESE.
THESE ARE MORE SAFE BECAUSE
THEY'RE EXQUISITELY
TUMOR-SPECIFIC AND THEY'RE NOT
SUBJECT TO CENTRAL TOLERANCE.
THERE IS REPRODUCIBLE DATA
SHOWING THAT THERE'S A
CORRELATION BETWEEN
ADMINISTERING HIGHER FREQUENCY
OF NEOANTIGEN REACTIVE TIL AND
HAVING A MORE LIKELY BETTER
RESPONSE TO TIL THERAPY AND ALSO
MORE LIKELY RESPONSE TO IMMUNE
CHECKPOINT BLOCKADE.
IMPORTANTLY, A VAST MAJORITY OF
PATIENTS WILL PRESENT
NEOANTIGEN-SPECIFIC TIL SO BY
TARGETING THESE NEOANTIGENS,
WHAT COULD POTENTIALLY GENERATE
A THERAPY TARGETING A BROAD
PATIENT POPULATION, AND THERE IS
INCREASING AMOUNT OF EVIDENCE OF
NEED OWE ANTIGEN SPECIFIC TO
INDUCING ANTITUMOR RESPONSES IN
SELECTED CANCER PATIENTS.
AND MANY OF THESE, MUCH OF THIS
WORK WAS BEING DONE IN THE
SURGERY BRANCH WHILE HE WAS A
POSTDOC.
SO WHEN I WENT BACK TO BARCELONA
TO START MY GROUP I DECIDED FOR
ONE OF MY MAIN FOCUSES TO START
A CLINICAL TRIAL WITH NEOANTIGEN
SELECTED TIL VERY MUCH INSPIRED
BY THE WORK I SAW HERE IN THE
SURGERY BRANCH.
SO ON ONE SIDE, WHAT WE DO, WE
RECENTLY STARTED THIS CLINICAL
TRIAL AND ON ONE SIDE, WE EXPAND
THE TIL FROM FRAGMENTS, WE THEN
SEQUENCE THE TUMOR, IDENTIFY
CANDIDATE MUTATIONS, WE EITHER
CLONE THESE IN THE FORM OF
TANDEM MINI GENES OR WE SIPT
SIZE LONG PEPTIDES AND WE USE
EITHER THESE RNA ENCODING FOR
TANDEM GENES OR PEPTIDES TO
PULSE OR EXPRESS THESE CANDIDATE
NEOANTIGENS IN AUTOLOGOUS
PRESENTING CELLS WHICH ARE USED
AS TARGETS, AND IF AND WHEN WE
FIND THESE NEOANTIGEN REACTIVE
TIL, WE SELECT THE TIL
CONTAINING THE REACTIVITY AND
FURTHER EXPAND FOR PATIENT
TREATMENT.
NOW IT BEEN A VERY STEEP
LEARNING CURVE, I'M NOT A
CLINICIAN AND THERE STILL IS
SOME CHALLENGES BUT I'M REALLY
EXCITED TO SEE THIS HAPPENING IN
BARCELONA.
THANKS FOR THE ENORMOUS TEAM OF
PEOPLE AND A LOT OF
COLLABORATIONS.
NA TO THESE SELECTED TIL, WE'RE
ALSO EXPLORING THE USE OF
NON-SELECTED TIL AND T CELLS
FROM PERIPHERAL BLOOD.
BUT TODAY I'M REALLY EXCITED TO
TELL BUT SLIGHTLY MORE BASIC
QUESTION THAT WE'RE TRYING TO
ADDRESS.
WE HAVE ESTABLISHED SEVERAL
METHODS THAT WE USE, THEY'RE
COMPLIMENTARY APPROACHES WE USE
IN PARALLEL TO INVESTIGATE THE
SPECIFIC TUMOR ANTIGENS THAT ARE
TARGETED BY TIL.
WE ROUTINELY GET TUMOR AND
MATCHED PERIPHERAL BLOOD WITH
PATIENTS, DIFFERENT CANCER
TYPES, AND WE ATTEMPT TO
GENERATE AN AUTOLOGOUS TUMOR
CELL LINE OR AN ORGANOID, AND IN
PARALLEL WE EXPAND TILES BUT WE
ALSO ISOLATE A SPECIFIC -- WHICH
WE KNOW ARE ENRICHED IN TUMOR
REACTIVITY.
IN ORDER TO IDENTIFY WHAT THESE
TIL RECOGNIZE, WE OF COURSE WANT
TO INCLUDE THE NEOANTIGENS AND
CANDIDATE ANTIGENS SO WE PERFORM
NORMAL DNA BUT ALSO TUMOR DNA
SEQUENCING TO IDENTIFY THESE
MUTATIONS, WE USE THIS
PERSONALIZED SCREENING APPROACH
WHICH I ALLUDED TO IN THE
PREVIOUS SLIDE, BUT IN ADDITION
TO INCLUDING THE NEOANTIGENS, WE
FREQUENTLY ALSO CLONE CANCER
GERMLINE ANTIGENS WHICH WE FIND
ARE EXPRESSED AT RNA LEVEL IN
THE TUMOR BIOPSIES BUT ALSO
TISSUE DIFFERENTIATION ANTIGEN
SO WE'RE NOT LIMITING OURSELVES
TO NEOANTIGENS ONLY IN THIS MORE
EXPLORATORY WORK.
WE FREQUENTLY USE HL.
A BINDING PREDICTION
SPECIFICALLY WHEN PATIENTS HAVE
A VERY LARGE NUMBER OF SOMATIC
MUTATIONS AND WE HAVE ALSO
ESTABLISHED OUR OWN
IMMUNOPEPTIDE OMICS PLATFORM
WHICH ENABLES US TO I MAW KNOW
PRECIPITATE THE -- ON CANCER
CELL LINES THAT WE GENERATE, AND
TO IDENTIFY THE SPECIFIC
PEPTIDES EITHER CONTAINING
MUTATIONS OR DERIVED FROM CANCER
GERMLINE ANTIGENS OR TISSUE
DIFFERENTIATION ANTIGENS.
SO THESE APPROACHES ARE
COMPLEMENTARY.
AND WE USE THEM ALL IN PARALLEL.
WE EITHER -- RNA ENCODING FOR
THESE OR WE SYNTHESIZE THE
PEPTIDES, FINALLY, TO SCREEN AND
TEST WHETHER THESE TILES THAT
WERE EXPANDED OR T CELLS FROM
BLOOD THAT CAN RECOGNIZE ANY OF
THESE ANTIGENS.
I'M GOING TO SHOW YOU DATA FROM
THREE PATIENTS WE SCREENED USING
THESE THREE PARALLEL APPROACHES.
THIS IS THE REACTIVITY TO THE
AUTOLOGOUS CELL LINE OF SEVERAL
TIL THAT WERE EXPANDED FROM A
METASTATIC MELANOMA PATIENT.
WE CAN SEE THERE'S VARIABLE
FREQUENCY OF TUMOR RECOGNITION
BASED ON 41BB EXPRESSION AND
THIS IS ONLY LIMITED TO
CD8 RESPONSES FOR THE MOMENT.
SO WE TOOK THESE TILES AND
SCREENED FOR ALL THESE
NEOANTIGENS AND ALSO
TUMOR-ASSOCIATED ANTIGEN
RECOGNITION.
WHAT WE FOUND IS THAT WE COULD
EXPLAIN, FOR EXAMPLE, A VERY
DOMINANT T-CELL RESPONSE WAS
PRESENCE IN THIS TIL FRAGMENT
THREE.
ALSO SOME NEOANTIGEN REACTIVITY
IN TIL 5 AND NOT VERY DOMINANT.
AND A FAIRLY DOMINANT POPULATION
TARGETING -- 5.
BUT WHAT WE ALSO OBSERVED, IN
FACT, IS THAT MANY OF THESE
TILs, DESPITE ALL THIS
EXTENSIVE SCREENING, WE STILL
DIDN'T KNOW WHAT TUMOR ANTIGENS
THEY RECOGNIZE.
THEY CLEARLY RECOGNIZED THE
CANCER CELLS, NOT NON-MATCHED
MELANOMA CELL LINES, BUT WE
COULDN'T STILL KNOW WHAT THEY
WERE RECOGNIZING.
AND THIS HAS BEEN THE
OBSERVATION OVER MANY PATIENTS.
SO THESE ARE TWO OTHER PATIENTS
WITH HEAD AND NECK CANCER.
HE WITH EXPANDED TIL, MANY OF
THEM CAN RECOGNIZE TUMOR TO A
DIFFERENT EXTENT.
WE SCREEN FOR NEOANTIGEN
RECOGNITION.
WE CAN CERTAINLY FIND THESE.
THIS IS WHAT MAKES IT ROW U.P.S.
AND WE CAN NOW POTENTIALLY
EXPLORE THIS TOOL TO TREAT
PATIENTS.
HOWEVER, AGAIN, MANY OF THESE
TIL RECOGNIZE CANCER AS WE STILL
DON'T KNOW WHAT THEY RECOGNIZE.
SO THIS BRINGS US BACK TO THE
QUESTION, WHAT ADDITIONAL
ANTIGENS CAN' LIS CAN ELICIT REN
CANCER PATIENTS?
WHEN WE MADE THIS O OBSERVATION
THERE WERE A GROWING NUMBER OF
ARTICLES POINTING TOWARDS
NON-CANONICAL ANTIGENS, ALSO
KNOWN AS CRYPTIC ANTIGENS OR
ANTIGENS DERIVED FROM DARK
MATTER.
THE FACT IS THAT MOST OF THE
CURRENT EFFORTS OF TUMOR ANTIGEN
DISCOVERY FOCUS ON TESTING THE
IMMUNOGENICITY OF ANNOTATED
PROTEINS WHICH REPRESENT
APPROXIMATELY 2% OF THE ENTIRE
GENOME.
HOWEVER, IT IS NOW KNOWN THAT
UNDER CERTAIN CIRCUMSTANCES, UP
TO 75% OF THE GENOME CAN BE
TRANSCRIBED AND POTENTIALLY
TRANSLATED.
AND THERE IS EVIDENCE OF SOME
LITERATURE OF T-CELL RESPONSES
TARGETING DIFFERENT TYPES OF
EPITOPES DERIVED FROM DARK
MATTER SUCH AS TRANSPOSABLE
ELEMENTS, ALTERNATIVE RNA
SPLICING, RNA TRANSLATION AND
PEPTIDE SPLICING.
MUCH OF THIS EVIDENCE WAS
GENERATED IN THE 1990s, USING
THE CDNA TUMOR LIBRARIES.
BUT THERE WAS EMERGING DATA
SUPPORTING THAT HLA ANALYSIS
COULD LEAD TO THE MORE
SYSTEMATIC IDENTIFICATION OF
THESE CRYPTIC EPITOPES IN A MORE
OR LESS PERSONALIZED FASHION
EITHER USING BIOPSIES OR CANCER
CELL LINES.
SO WE REASONED THAT IF ABERRANT
TRANSLATION HAPPENS EXCLUSIVELY
IN CANCER CELLS, THIS COULD LEAD
TO THE PRESENTATION OF
IMMUNOGENIC PEPTIDES THAT COULD
POTENTIALLY EXPAND THE LANDSCAPE
OF TARGETABLE ANTIGENS FOR
CANCER IMMUNOTHERAPY.
AND THIS BECAME THE QUESTION OF
A VERY TALENTED GRADUATE STUDENT
THAT I HAD IN MY LAB, SHE'S NOW
A POSTDOC, AND SO WHAT SHE DID
IS SHE PERFORMED IMMUNOPEPTIDE
OMICS OF CELL LINES OF DIFFERENT
ORIGIN.
I WILL EXPLAIN LATER THE ORIGIN.
FOLLOWING THIS AND IN
COLLABORATION, WE DECIDED TO USE
HIS PIPELINE PEPTIDE PRISSMM,
IT'S A PROTEOGENOMICS PIPELINE
WHICH ENABLE US ON ONE HAND TO
IDENTIFY PEPTIDES THAT MAP TO
THE ANNOTATED PROTEINS IN THE
GENOME, BUT IT CAN ALSO MAP
PEPTIDES TO THE THREE FRAME
TRANSCRIPTOME, SIX FRAME GENOME,
AND ALSO INCLUDING MUTATIONS
IDENTIFIED THROUGH WHOLE EXOME
SEQUENCING.
SO USING THIS APPROACH, ONE CAN
IDENTIFY PEPTIDES THAT ARE
MAPPING TO FIVE PRIME UTRs,
THREE PRIME UTRs, OFF-FRAME,
INTRONIC REGIONS, INTERGENIC AND
ALSO POTENTIALLY NON-CONTAINING
MUTATIONS.
SO SUFFICE TO SAY WE USED THIS
APPROACH TO IDENTIFY THE
CANONICAL PEPTIDES, WHICH WERE
EITHER CONTAINING MUTATIONS,
CANCER GERMLINE ANTIGENS OR
TUMOR-ASSOCIATED ANTIGENS, BUT
ALSO THESE NON-CANONICAL
ANTIGENS.
BECAUSE SOME OF THESE
NON-CANONICAL ANTIGENS CAN BE
POTENTIALLY TRANSLATED IN ALSO
HEALTHY TISSUE, WE USE THIS HLA
LIGAND ATLAS TO FILTER OUT
WHATEVER PEPTIDES THAT COULD
POTENTIALLY BE ALSO FOUND IN
THESE LIGAND -- 2, FINALLY GET A
LIST OF MORE TUMOR PREFERENTIAL
NON-CANNON CAN ANTIGENS
PREFERENTIALLY PRESENTED IN
CANCER CELLS.
SO VERY BRIEFLY, I WANT TO SHOW
YOU THE DATA.
SO ACROSS NINE DIFFERENT CANCER
CELL LINES, YOU CAN SEE WE HAVE
FOUR TUMORS, TWO HEAD AND NECK,
THREE MELANOMAS, WE WERE ABLE TO
IDENTIFY NON-CANONICAL PEPTIDES
IN ALL NINE CANCER CELL LINES.
AND THEY REPRESENTED A
RELATIVELY SMALL FRACTION OF ALL
THE PEPTIDES THAT WE WERE EABL
TO ELUTE FROM THESE CELL LINES.
USING THIS FILTER, FILTERING OUT
ALL THE PEPTIDES NON-CANONICAL
PEPTIDES THAT COULD BE DETECTED
IN THIS HLA LIGAND ATLAS OF
HEALTHY TISSUES, WE THEN
OBTAINED A MORE NARROW LIST OF
NON-CANONICAL TUMOR LIGANDS.
THESE ARE POTENTIALLY
PREFERENTIALLY PRESENTED BY
CANCER CELLS.
SO WE DECIDED TO STUDY THIS IN
MORE DETAIL.
WHEN WE LOOKED AT WHAT TYPE OF
NON-CANONICAL ANTIGENS THESE
DERIVE FROM, WHAT WE FOUND IS
THAT THEY MAINLY DERIVE FROM
FIVE-TIME UTR OR ALSO
ALTERNATIVE OPEN READING FRAMES,
NON-CODING RNA, AND LESS
FREQUENTLY TO THREE PRIME
UTRs, INTRONIC OR INTERGENIC
REGION.
AND WE WERE ONLY ABLE TO
IDENTIFY ONE SINGLE PEPTIDE THAT
WAS CRYPTIC AND CONTAINED A
MUTATION.
WE THOUGHT POTENTIALLY THIS ONE
COULD BE REALLY INTERESTING TO
TEST.
ON THE RIGHT WHAT YOU CAN SEE IS
THE COMPARISON IN TERMS OF THE
NUMBER OF PEPTIDES THAT WE
IDENTIFIED FROM EACH OF THE --
WE HAVE MUTATED PEPTIDES ON THE
TOP, CANCER GERMLINE MELANOMA
ASSOCIATED AND THE NON-CANONICAL
TUMOR LIGANDS.
AS YOU CAN SEE WHAT WE FOUND IS
THAT INDEED THESE NON-CANONICAL
TUMOR LIGANDS APPEARED TO BE
QUITE AN ABUNDANT SOURCE OF
ANTIGENS.
BUT YET WE NEEDED TO TEST
WHETHER THEY WERE IMMUNOGENIC.
THIS WAS THE MAIN QUESTION THAT
WE INITIALLY SET TO RESPOND.
SO TO ADDRESS THIS, WHAT WE DID
IS WE SYNTHESIZED ALL THE
PEPTIDES SHOWN HERE IN THIS
GRAVEL ONGRAPH ON THE RIGHT.
WHAT WE AIMED TO DO IS TEST THIS
IN A PERSONALIZED FASHION.
SO FOR EXAMPLE THE PEPTIDES FROM
MEL 3 WOULD ONLY BE TESTED FOR
THAT PATIENT.
SO THE TILES WHICH WE HAD TILs
WERE RECOGNIZING THE AUTOLOGOUS
SELL LINE WERE CO-CULTURED WITH
THE CELL LINES DERIVED FROM THIS
MELANOMA CELL LINE.
AND THE FINDINGS IN FACT WERE
QUITE INTRIGUING I MUST SAY.
SO I'M GOING TO FOCUS, AND I
WISH I COULD HAVE A POINTER?
SO FOCUSING HERE ON ONE OF THE
PATIENTS WE STUDIED IN DETAIL,
YOU CAN SEE THAT FOR THIS
PATIENT, WE SCREENED SEVERAL TIL
POPULATIONS THAT WE HAD THAT WE
KNEW RECOGNIZED THE AUTOLOGOUS
TUMOR CELL LINE.
PARTICULARLY IN THIS PATIENT WE
IDENTIFIED 215 NON-CANONICAL
TUMOR LIGANDS, SEVERAL TUMOR
ASSOCIATED CANCER GERMLINE AND
THREE MUTATED PEPTIDES.
AND AS YOU CAN SEE, WE FOUND
THAT TWO OUT OF THE THREE
MUTATED PEPTIDES WERE
IMMUNOGENIC.
TWO OUT OF THE 22 TUMOR
ASSOCIATED ANTIGENS WERE
IMMUNOGENIC BUT NON-OF THE 215
NON-CANONICAL TUMOR LIGANDS WE
IDENTIFIED WERE IMMUNOGENIC,
UNFORTUNATELY.
AND IN FACT, GLOBALLY, WE DID --
WE WERE NOT ABLE TO DETECT ANY
REACTIVITY TO ANY OF THE
NON-CANONICAL TUMOR LIGANDS
IDENTIFIED.
I MUST SAY THAT THIS WAS QUITE
SURPRISING.
WE HAD IN FACT VALIDATED THIS
PEPTIDE SO WE'RE PRETTY
CONFIDENT THAT THESE ARE
ACTUALLY PRESENTED ON THE CELL
SURFACE OF THE CANCER -- WE WERE
QUITE STRUCK EVEN THOUGH THE
LITERATURE POINTING TOWARDS
THESE ANDROGENS.
AND YET WE STILL WANTED TO KNOW,
WELL, YOU KNOW, POTENTIALLY THEY
ARE NOT BEING RECOGNIZED IN
CANCER PATIENTS NAYTIVELY, BUT
COULD WE IDENTIFY T CELLS AND
TCRs TARGETING NON-CANONICAL
TUMOR LIGANDS.
SO -- WITH A LIST OF
NON-CANONICAL TUMOR LIGANDS THAT
WERE IDENTIFIED AND ELUTED FROM
HLA A1101.
FOLLOWING THIS DESENSITIZATION,
DESENSITIZATION -- TARGETING
THREE DISTINCT NON-CANONICAL
TUMOR LIGANDS SHOWN HERE, ONE
DERIVING FROM FIVE PRIME UTR --
13, ONE FROM A NON-CODING RNA
AND ANOTHER ONE FROM A SECOND
FIVE PRIME UTR.
AND AS SHOWN ON THE RIGHT, THE
HLA RESTRICTION OF THESE
PEPTIDES WAS PROVEN TO BE FOR
TWO OF THEM HLA11 RESTRICT AND
FOR ONE OF THEM, HLA6801.
THE MAIN QUESTION WE HAD AFTER
SEEING THESE REACTIVITIES WAS
WHETHER THESE T-CELL POPULATIONS
WERE CAPABLE OF RECOGNIZING
THESE PEPTIDES NATURALLY
EXPRESSED AND PRESENTED BY
CANCER CELLS WHERE THEY WERE
INITIALLY IDENTIFIED.
AND SO WE PERFORMED THE
EXPERIMENTS, AND THE FINDINGS
WERE REALLY INTERESTING.
INDEED ONE -- THE CORRECT HLA
RESTRICTION ELEMENTS, YOU CAN
SEE THAT THESE T CELLS RESPOND
TO THE PEPTIDE WELL.
THEY DO NOT RESPOND TO MELANOMA
CELL LINE WHERE THESE ANTIGENS
WERE NOT IDENTIFIED.
EVEN WHEN YOU TRANSFECT HLA A11
OR A6801.
BUT THEY DO NATURALLY PRESENT
AND NATURALLY RECOGNIZE MEL 3.
SO LONG AS THE HLA RESTRICTION
ELEMENT IS ALSO PRESENT.
AND THIS CELL LINE IS NAYTIVELY
A1101 POSITIVE.
SO WE NEXT WANTED TO KNOW ONE OF
THE ADVANTAGES POTENTIALLY OF
TARGETING THESE NON-CANONICAL
CRYPTIC PEPTIDES THESE ARE NOT
PRIVATE SO POTENTIALLY BY
TARGETING THIS ONE COULD TREAT A
LARGER PATIENT POPULATION SO
LONG AS THIS IS PROCESSED,
PRESENTED AND RECOGNIZED BY THE
T CELLS.
AND ON DIFFERENT CELL LINES.
SO WHAT WE DID IS WE CLONED THE
TCR GENERATED TCR TRANSDUCED T
CELLS TARGETING -- JUST SHOWING
THE RESULTS FOR ONE OF THEM,
TARGETING 5 PRIME UTR OF HOC13.
AS YOU CAN SEE WE CO-CULTURED
THESE CELLS WITH A GOOD NUMBER
OF CANCER CELL LINES THAT WE
GENERATED OVER THE YEARS FROM
DIFFERENT ORIGIN GYNECOLOGICAL
HEAD AND NECK MELANOMA GI AND
RENAL AND LUNG, AND OBSERVED IN
THE -- WHAT WE OBSERVED WAS THEY
WERE ABLE TO RECOGNIZE ONLY THE
MELANOMA CELL LINES THAT WEREN'T
NATURALLY EXPRESSING HLA A1101
WHICH WAS THE RESTRICTION
ELEMENT FOR THE TCR.
NONETHELESS WHEN WE INTRODUCED
HLA SN 1101 INTO ALL THE REST OF
THE CANCER CELL LINES, WE BEGAN
TO SEE SOME RECOGNITION OF GI
CANCER OR GYNECOLOGICAL CANCER,
AGAIN POTENTIALLY SUGGESTING
THAT THESE CRYPTIC PEPTIDES
COULD BE TARGETABLE IN OTHER
CANCER TYPES.
AND NOT WITH A6801.
THIS WAS REALLY INTERESTING, WE
WERE STILL SLIGHTLY WORRIED THAT
WE DIDN'T KNOW HOW
TUMOR-SPECIFIC THESE ANTIGENS
WERE.
TO ADDRESS THIS -- ADDRESSING
THIS IS NOT THAT SIMPLE BECAUSE
THIS ABERRANT TRANSLATION CANNOT
BE DETECTED AT RNA SEQ LEVELS SO
WE CANNOT USE RNA SEQ TO SEE
WHETHER CELL LINES ARE POSITIVE
OR NEGATIVE FOR OUR TARGETS.
SO WE USED A SLIGHTLY DIFFERENT
APPROACH.
WE PURCHASED HEALTHY CELLS.
THESE ARE RENAL FIBROBLASTS,
EPITHELIAL CELLS, CARDIAC
MYOCYTES OR MELANOCYTES.
WHAT WE SEE IS THAT, IN FACT,
THESE TCR TRANSDUCED T CELLS CAN
RECOGNIZE THESE CELL LINES SO
LONG AS THE HLA RESTRICTION
ELEMENT IS THERE, IT CAN EVEN
INCREASE WHEN YOU FURTHER
ENHANCE THE EXPRESSION OF HLA
RESTRICTION ELEMENT, BUT IT DOES
NOT DISPLAY RECOGNITION OF ANY
OF THE HEALTHY CELLS.
NOW WE'RE ONLY TESTING A FEW
NUMBER OF NORMAL HEALTHY CELLS
AND THIS MAY BE A LIMITATION,
BUT WE DID HAVE A POSITIVE
CONTROL OF A TCR THAT WE KNOW
CAN LEAD TO AUTOIMMUNE TOXICITY
WHICH IS THE MART 1 CELLS AND
THESE ARE RESTRICTED -- NOT ONLY
THEY RECOGNIZE MELANOMA CELL
LINES THAT ARE TRANSDUCED WITH
HLA -- AND SURPRISINGLY TO A
LOWER EXTENT CARDIAC MYOCYTES.
SO I THINK THIS -- IN SUMMARY, I
FIND THAT THESE NON-CANONICAL
TUMOR LIGANDS CAN BE IMMUNOGENIC
AND SHARED ACROSS CANCER TYPES
AND APPEAR TO BE PREFERENTIALLY
PRESENTED BY CANCER CELLS.
I THINK AT THIS POINT WE STILL
HAVE TO EVALUATE HOW
TUMOR-SPECIFIC THEY ARE AND ALSO
WE HAVE TO FIND WAYS POTENTIALLY
HOW TO LOGISTICALLY SELECT THESE
PATIENTS IF WE WANT TO PUSH
FURTHER WITH THE TCR TRANSDUCED
APPROACH.
WE FOUND THAT ABERRANT
TRANSLATION DID NOT EXPLAIN THE
RESPONSES AND THIS BEGS THE
QUESTION WHY DIDN'T THESE
CRYPTIC PEPTIDES ELICIT A T-CELL
RESPONSE IN CANCER PATIENTS.
WE HAVE SOME POTENTIAL TECHNICAL
EXPLANATIONS WHICH WE HAVE MORE
OR LESS RULED OUT SINCE WE THINK
THOSE SHOULD AFFECT OTHER
NEOANTIGENS AND OTHER
TUMOR-ASSOCIATED ANTIGENS.
BUT WE FEEL THAT TWO PLAUSIBLE
HYPOTHESES FOR THAT IS THAT WE
BELIEVE THESE PEPTIDES ARE
POTENTIALLY VERY EFFECTIVELY
PRESENTED ON -- OF CANCER CELLS
BUT LESS EFFECTIVELY CROSS
PRESENTED POTENTIALLY LEADING TO
INEFFECTIVE PRIMING OF T-CELL
RESPONSES.
NOW THIS ALTHOUGH COULD BE A
DISADVANTAGE IN TERMS THAT
THEY'RE NOT NATURALLY
IMMUNOGENIC COULD BE AN
ADVANTAGE, AS THEY HAVE NOT BEEN
SO IMMUNOEDITED.
SO THIS IS SOMETHING THAT
POTENTIALLY WE NEED TO STUDY
MORE IN DEPTH.
THERE'S A SECOND HYPOTHESIS,
THOUGH, WHICH IS INDEED THESE
ARE NOT SO TUMOR-SPECIFIC,
POTENTIALLY THIS COULD BE
EXPRESSED AND PRESENT BID KIEEDY
THYMIC -- DURING THYMIC
DEVELOPMENT.
FINALLY WE ARE STILL MISSING A
CONSIDERABLE FRACTION OF
ANTIGENS TARGETED BY T CELLS IN
CANCER PATIENTS AND WE ARE STILL
DRIVEN TO CONTINUE TO ADDRESS
THIS QUESTION.
WE THINK FUSIONS, ALTERNATIVE
SPLICING, INTRON RETENTION,
STRUCTURAL VARIANTS AND
CERTAINLY TUMOR ASSOCIATED AND
TISSUE DIFFERENTIATION AND
CANCER GERMLINE ANTIGENS.
HOWEVER THIS LIST OF ANTIGENS IS
EVER-GROWING AND WE FEEL THAT
THIS PROTEOMICS APPROACH IS NOT
SENSITIVE ENOUGH, AND WE ARE IN
NEED OF MORE HIGH-THROUGHPUT
STRATEGIES TO -- SUCH A LONG
LIST OF PEPTIDES.
AND FOR THIS REASON, WE ARE
MOVING TOWARDS A MORE
HIGH-THROUGHPUT SCREENING OF
USING DNA BASED TUMOR LIBRARIES.
THIS IS SPEARHEADED BY A POSE
DPOSTDOCAND GRADUATE STUDENT INB
WHO HAVE COME UP WITH THIS TITLE
OF UNBIASED PERSONALIZED
SCREENING OF CANDIDATE ANTIGEN
LIBRARIES.
WHEREBY WE INTEND TO
IMMORTGAGIZE B CELLS AND EXPRESS
AN ANTIGEN LIBRARY WHERE EACH B
CELL IS EXPRESSING A DIFFERENT
CANDIDATE ANTIGEN AND YOU CAN
HAVE UP TO TENS OF THOUSANDS OF
ANTIGENS AND CO-CULTURE T CELLS
OR TCRs OR TCR LIBRARIES WITH
THESE CANDIDATE ANTIGENS,
ISOLATE THOSE THAT HAVE BEEN
RECOGNIZED BY T CELLS TO GZB
REPORTER AND FINALLY SEQUENCING
THE RECOGNIZED ANTIGENS TO
DISCOVER NOVEL PEPTIDES.
THIS IS ONGOING WORK, AND WITH
THAT, I WILL END WITH THE
ACKNOWLEDGMENTS.
I'D LIKE TO THANK THE PEOPLE
FROM MY GROUP THAT HAVE
CONTRIBUTED TO THIS WORK.
ALL THE COLLABORATORS, THE
FUNDING AI AGENCIES AND VERY
ESPECIALLY ALL PATIENTS THAT
CONTRIBUTED.
THANK YOU VERY MUCH.
[APPLAUSE]
>> THANK YOU, DR. GROS.
OUR FINAL SPEAKER FOR THIS
SESSION WILL BE DR. KEN-ICHI
HANADA, STAFF SCIENTIST FROM THE
SURGE ROW BRANCH.
SURGERY BRANCH.
>> SO MY -- I HAVE TO USE MY
LAPTOP.
GIVE ME A SECOND TO SET IT UP.
SO WHILE WE'RE WAITING I'D LIKE
TO SHARE MY EXPERIENCE THROUGH
WHICH I CAME TO UNDERSTAND WHAT
THE SURGERY BRANCH IS ABOUT.
I WAS FORTUNATE ENOUGH TO
IDENTIFY POST TRANSLATIONALLY
SPLICED PEPTIDE.
IT TOOK ME MORE THAN THREE
YEARS, FOUR YEARS, AND FINALLY
FROM FOUND THE EPITOPE, I WAS FF
JOY.
AND THEN AT THE MEETING I
PRESENTED THE DATA TO
DR. ROSENBERG.
SO WHEN I WAS PRESENTING THE
DATA, I HAD THIS BURNING
QUESTION, OKAY, I FOUND THIS
EPITOPE, BUT HOW IS THIS
HAPPENING?
WHAT IS THE MECHANISM?
THAT WAS MY BURNING QUESTION
THAT GET ME AWAKE.
SO WHEN I PRESENTED THE DATA TO
DR. ROSENBERG, I EXPECTED AN
ENCOURAGING COMMENT LIKE, OKAY,
NOW YOU WANT TO FIND THE
MECHANISM, WHAT WILL BE YOUR
APPROACH?
THAT KIND OF COMMENT, I
EXPECTED.
BUT DR. ROSENBERG'S COMMENT WAS
VERY DIFFERENT.
HE WAS LIKE, OKAY, YOU MADE A
GREAT DISCOVERY.
IT'S VERY INTERESTING.
SO OTHER PEOPLE WILL WORK ON
THIS TOO.
SO YOU STOP IT.
SO YOU HAVE TO MOVE ON.
HE ALSO SAID THAT --
DR. ROSENBERG ALSO SAID WE HAVE
TO WORK ON THE QUESTIONS, WE
THINK IS IMPORTANT, BUT NO OTHER
PEOPLE FEEL SO IMPORTANT.
SO WE HAVE TO FOCUS OUR POWER,
RESEARCH POWER ON THAT KIND OF
QUESTION.
THAT WAS HIS COMMENT.
SO WHEN I HEARD THE COMMENT, AT
FIRST FOR A SECOND OR TWO, I WAS
LIKE, WHAT?
I WAS SO DISAPPOINTED FOR A
SECOND.
BUT YOU KNOW, IT DIDN'T TAKE
LONG TO REALLY UNDERSTAND, YEAH,
THAT'S -- YOU KNOW, THAT'S THE
REASON WHY -- OUR EXISTENCE, THE
BRANCH, THAT'S WHAT THE BRANCH
IS ABOUT, THAT'S WHAT
DR. ROSENBERG IS ABOUT.
HE HAS BEEN THE -- TO ME HE HAS
BEEN THE LIGHTHOUSE OR THE
BEACON FOR ALREADY 27 YEARS, AND
THEN EVERY DAY, I CAN COME IN
BEING EXCITED.
SO I FEEL IT A GIFT, AND
DIRECTLY THANK DR. ROSENBERG FOR
IT.
OKAY.
SO NOW, BY THE WAY, MY NAME IS
KEN-ICHI HANADA.
I'M A STAFF SCIENTIST AT
DR. JAMES YANG'S LABORATORY.
THTODAY'S TALK IS TITLED ANTITUR
MECHANISM MEDIATED BY CLASS II
MHC-RESTRICTED CD4 PLUS T CELLS.
NOW I'D LIKE TO START WITH AN
INTRODUCTION.
AS A SURGERY BRANCH, WE EMPLOY
THREE APPROACHES TO TREAT
PATIENTS USING -- CELL TRANSFER.
FIRST TUMOR INFILTRATING
LYMPHOSIGNS, TILs, WITH TUMOR
REACTIVITY.
SECOND APPROACH IS AUTOLOGOUS
LYMPHOCYTES ENGINEERS TO EXPRESS
PATIENT-SPECIFIC
NEOANTIGEN-REACTIVE T-CELL
RECEPTOR, SO-CALLED TCR
PROTOCOLS, AND THE THIRD WAY IS
OFF THE SHELF TCRs TARGETING
COMMON DRIVER MUTATION DERIVED
EPITOPES PRESENTED BY WIDELY
SHARED MHC MOLECULE.
DR. YANG PRESENTED THAT.
SO -- BUT IN CONDUCTING THESE
CLINICAL TRIALS, WE REALIZED
SOME PROBLEMS.
ONE PROBLEM IS A SIGNIFICANT
NUMBER OF PATIENTS HAVE
ALREADY -- HAVE ALREADY
EXPERIENCED MHC LOSS PRIOR TO
THE THERAPY AS A BRANCH.
SO THIS IS -- THESE PIE CHARTS
SHOW THE FREQUENCY OF -- LOSS AT
THE SURGERY BRANCH - BASED ON
WHOLE EXOME SEQUENCING.
THERE ARE SIX CANCER, AND
ROUGHLY -- SO THE RED PIE,
THAT'S A LOSS OF MHC LOSS.
BLUE IS INCONCLUSIVE, BECAUSE WE
ANALYZED THE -- USING TUMOR
TISSUE SO DEPENDING ON
CONTAMINATION OF THE -- CELLS,
MEASURING OR JUDGING JUDGMENT
LOH -- SO THERE ARE MANY
INCONCLUSIVE CASES.
BUT AFTER AT LEAST 20 TO 30%,
THIS IS A VERY -- BASED ON THE
VERY STRICT CRITERIA, SO 22, 30%
OF THE PATIENTS COME IN WITH MHC
LOSS.
SO THAT'S ONE BIG PROBLEM.
ON THE SECOND PROBLEM IS T-CELL
REACTIVITY CAN LEAD TO THE SLEK
THECOLLECTION OF TUMOR CELLS WIH
LOSS OF FUNCTION MUTATION --
LOSS OF FUNCTION MUTATIONS OR
LOSS OF MHC MOLECULES.
AND THEN THERE ARE EXAMPLES OF
THAT, SO IF I BRIEFLY MENTION
THESE EXAMPLES.
THE FIRST ONE WAS PUBLISHED BY
ERIC TRAN AND THEN THIS WAS A
CASE IN WHICH COLORECTAL CANCER
PATIENT WAS TREATED WITH KRAS --
AND THIS RESULTED IN THE
EMERGENCE OF RECURRENT TUMOR
WITH HLA-C0802 LOSS.
THE SECOND CASE IS A MORE RECENT
CASE, SO THIS PATIENT WAS
TREATED WITH AGAIN KRAS G12D
BUT -- RESTRICTED.
INITIALLY, SEVEN MONTHS, VERY
GOOD RESPONSE WAS SEEN.
BUT AFTER TWO MONTHS, THERE WAS
A SMALL RECURRENCE AND THIS
TUMOR HAD MUTATION IN THE
RESTRICTED MHC, LOSS OF
FUNCTION -- MUTATION MHC.
SO THIS, A BIG PROGRAM WITH
ATTACKING TUMOR WITH CD8 T
CELLS.
SO TODAY I'D LIKE TO TALK ABOUT
CD4 T CELLS AND THEN I'D LIKE TO
EXPLAIN WHY CD4 T CELLS CAN BE
IMPORTANT.
SO FIRST I'D LIKE TO EXPLAIN TO
YOU, DOES -- ANTITUMOR CD4
RESPONSE IS MORE COMPLICATED
THAN CD8 RESPONSE.
SO I WANT TO BRIEFLY EXPLAIN
ABOUT THAT.
THEN CD8 T CELLS, MOST OF THE
REACTIVITY IS BASED ON THE
DIRECT RECOGNITION OF TUMORS
THROUGH -- PRESENTING THE
EPITOPE.
VERY SIMILAR THING CAN HAPPEN
WITH CD4 TOO.
NOW IN THE CASE OF CD4, THE
ANTIGEN PROCESSING MECHANISM IS
SLIGHTLY DIFFERENT BUT AT THE
END OF THE DAY CD4 T CELLS CAN
REPRESENT AN EPITOPE PRESENT ON
THE SURFACE OF THE TUMOR, SO --
EXIST IN BOTH CASES.
THE MECHANISMS DOESN'T SEEM TO
EXIST ON CD8, CD4 UNIQUE
MECHANISM -- ANTITUMOR MECHANISM
BY CD4 T CELLS.
SO THIS HAS BEEN SHOWN MOUSE
MODEL.
THE CONCEPT CD4 T CELLS DON'T
NEED AN MHC EXPRESSION OF TUMOR
CELLS, IT'S NOT A NEW IDEA.
THERE WERE TWO MAJOR
PUBLICATIONS THAT SHOWED THAT
WORKED IN DEPTH ABOUT HOW THIS
ANTITUMOR MECHANISM BY CD4 IS
WORKING.
ACCORDING TO THEM, SO THE TUMOR
CELLS SECRETE ANTIGENS.
THE ANTIGEN IS TAKEN UP BY A PSM
C WHICH IS RECOGNIZED BY CD4.
THE CD4 T CELLS SECRETE
CYTOKINES SUCH AS INTERFERON KA
GAMMA -- MONOCYTES OR
NEUTROPHILS.
SO -- SECRETE EFFECTOR MOLECULES
SUCH ACINI TRICK OXIDE AND THAT
RESULTS IN KILLING OF THE TUMOR.
STILL, THEY DIDN'T HAVE A CLEAR
EXPLANATION ABOUT WHERE THE
TUMOR SPECIFICITY COMES FROM.
THEY DID NOT HAVE THAT KIND OF
EXPLANATION, OR BECAUSE OF THE
COMPLEXITY, WE STILL DON'T KNOW
IF THIS MECHANISM -- YOU CAN
POTENTIALLY TREAT -- TUMOR SO
IT'S A VERY -- BUT AT THIS POINT
WE DON'T KNOW IF IT CAN BE
TRANSLATABLE TO HUMAN.
SO I'M TRYING TO ANSWER THE
QUESTION IF IT IS TRANSLATABLE
TO HUMAN.
THAT'S ONE REASON WHY CD4 IS
IMPORTANT.
THE SECOND REASON CD4 CAN BE
IMPORTANT IS THAT THIS IS FROM
A -- BY MARIAH, SO IN THIS
STUDY, AS YOU MAY KNOW, IN THE
CASE OF MELANOMA, CD8 REACTIVITY
WAS OVERWHELMINGLY DOMINANT, BUT
IN THE CASE OF -- CANCER, IT'S
NOT THE CASE.
ACTUALLY CD4 IS MORE THAN CD8,
AND IN SOME CASES, I THINK ONLY
CD4 REACTIVITY, SO THAT MAKES --
DEPENDING ON THE CASE, IT
BECOMES AN IMPORTANT REACTIVITY.
AND THEN THIS IS IN THIS -- THIS
IS A CLINICAL TRIAL CONDUCTED AT
THE BRANCH RECENTLY.
IT'S A CLINICAL PROTOCOL THAT
TREATED THE GASTROINTESTINAL
CANCER USING TIL.
THEN FRANK LOWERY AND STEPHANIE
GOFF EXAMINED THE ASSOCIATION
BETWEEN CLINICAL RESPONSE AND
HIGHER CD4 T-CELL COUNTS.
IN THIS CASE, CD4 T-CELLS SHOWED
BETTER ASSOCIATION WITH PARTIAL
RESPONSE.
SO I GUESS WE STILL DON'T KNOW
THE EXACT MECHANISM, BUT IF THIS
IS THE CASE, AGAIN, CD4 T CELLS
MAY BE A -- AND THE LAST ONE IS
ACCUMULATING SUCCESSFUL --
CLINICAL RESPONSES FOUND WITH
CD4 T CELLS.
THE FIRST EXAMPLE IS THE PAPER
IN WHICH THEY TREATED -- USING
THE -- CD4 T CELLS.
IT WAS A SINK SINGLE CASE BUT S
RESULTED IN THE COMPLETE
RESPONSE.
AND THEN THE SECOND EXAMPLE IS
AGAIN ERIC TRAN'S PUBLICATION,
IN THIS CASE -- REACTIVE TIL
RESULTED IN CLINICAL RESPONSES.
AND THEN THE THIRD EXAMPLE IS
THIS IS A TCR TRANSDUCTION STUDY
USING THE -- RECEPTOR -- AND IN
THIS CASE TWO COMPLETE RESPONSES
AGAINST CERVICAL CANCER AND --
CANCER AND THEN TWO PARTIAL
RESPONSES AGAINST ESOPHAGEAL
CANCER AND THEN OSTEOSARCOMA
AMONG 17 PATIENTS TREATED.
SO THIS IS AN EXAMPLE OF THE
COMPLETE RESPONSE WITH --
CANCER.
THE LEFT SIDE SCAN IS FROM
AUGUST TO -- AND THE RIGHT SIDE
IS FROM 2019.
YOU CAN SEE THE MULTIPLE
VISCERAL METASTASES DISAPPEARED
AFTER THIS THERAPY.
SO CD4 THERAPY CAN WORK.
SO BASED ON THESE FINDINGS, I
STARTED MY EXPERIMENTS.
SOME EXPERIMENTS I'M GOING TO
PRESENT ARE ALREADY PUBLISHED,
BUT I WANTED TO CONFIRM TO SEE
HOW IT GOES.
SO I USE THIS MOUSE MODEL, AND T
CELLS ARE -- REACTIVE CD4 T
CELLS RESTRICTED BY -- AND THEN
ISOLATED T CELLS FROM THE TCR
TRANSGENIC MICE OR USING
RETROVIRUS TRANSDUCE THE T-CELL
RECEPTOR TO CD4 SPLENOCYTES.
THIS TRANSDUCTION EXPERIMENT I
PERFORM IS BECAUSE THAT'S WHAT
WE DO WHEN WE DO THE CLINICAL
TREATMENT, WE TRANSDUCE TCR BY
RETROVIRUS, SO I WANTED TO MIMIC
THAT CLINICAL TRIAL SO I TOOK
THIS WAY.
FOR THE TUMOR, B16-F10 OR
B16-F10 WITH H -- LOSS BY
CRISPR.
SO B16-F10 RESULTING GAMMA
TREATMENT DOES NOT EXPRESS --
MHC BUT WITH INTERFERON
TREATMENT -- AND IT DOESN'T
HAPPEN IN THE CASE OF THE CRISPR
KNOCKED OUT B16 AND THEN ON THE
RIGHT SIDE I'M SHOWING DIRECT
RECOGNITION OF THE TUMOR BY THE
T-CELL, SO THIS T-CELL CAN'T
DIRECTLY RECOGNIZE THE -- THE
RESTRICTION ELEMENT AND WHICH
DOES NOT EXIST IN THE CASE OF
THE CRISPR KNOCKOUT.
SO USING THESE I PERFORM THE
FOLLOWING EXPERIMENT.
SO THE FIRST IN VIVO MODEL I
PERFORMED WAS FIRST I INJECTED
B16 OR H-2A -- KNOCKOUT B16 TO
MICE AND THEN WAITED FOR 10 OR
11 DAYS UNTIL TUMOR IS
ESTABLISHED.
WE DID A WHOLE BODY IRRADIATION
AND ON DAY ZERO, I GAVE TYRP1 T
CELLS IV IN CONJUNCTION WITH
ANTI-OX40 ANTIBODY.
SO THIS IS A MODEL BY HIRSCHO
RN'S GROUP.
THIS MODEL HAS A STRENGTH, THE
POTENTIAL FOR -- AGONIST
AVAILABLE USED IN CLINICAL
TRIALS AND HAVE SHOWN SAFETY.
AND THEN THIS METHOD DOES NOT
NEED VACCINATION.
SO VACCINATION CAN BE LIMITATION
IN THE CLINICAL SETTING BECAUSE
OFTENTIMES THE MUTATIONS ARE
SPECIFIC TO PATIENTS.
SO IF YOU HAVE TO PREPARE
PATIENT-SPECIFIC VACCINES, IT
BECOMES A HEAVY BURDEN TO OUR
SITE.
BUT IN THIS CASE, IT'S NOT
NEEDED.
SO THE TOP -- AND THEN THE -- IN
BOTH CASES, BOTH TUMORS GROW
RAPIDLY, AND THEN WE HAVE TO
EUTHANIZE THE MICE.
AND THEN IF YOU USE THE T-CELL
AND THEN ANTIOX40, IN BOTH
CASES, THIS CASE, THERE IS NO
RESTRICTIONAL ELEMENT -- SO IN
THIS CASE THIS RESPONSE SEEMS TO
BE HAPPENING RESULT OF DIRECT
CYTOTOXICITY.
FOR THIS KIND OF RESPONSE TO
HAPPEN, YOU NEED BOTH T CELLS
AND THE ANTIOX40 BECAUSE
ANTIOX40 ANTIBODY ALONE DOES NOT
DO IT OR TYRP1 T CELLS DOES NOT
DO THAT EITHER.
SO JUST A QUICK SLIDE SHOWING IT
IS NOT LINKED TO -- IN THIS CASE
I USE OVER TUMOR CELLS B16-F10
OR F10 WITH OVA AND THEN THESE
MICE WERE TREATED WITH OT-II T
CELLS IN THIS CASE AND IT WASN'T
AS STRONG AS TYRP BUT YOU SEE
OBVIOUS YOU CAN ACHIEVE COMPLETE
RESPONSE SO IT'S NOT LIMITED TO
TYRP1.
IN THIS EXPERIMENT, I EXAMINE
THIS RESPONSE DEPENDENCY ON
THOSE HOST T CELLS AND B CELLS
BY UTILIZING RAD KNOCKOUT MICE.
THE LEFT MOST IS B16-F10 SO THIS
IS B16-F10 GIVEN TO NORMAL MICE.
YOU SEE GOOD RESPONSE.
THEN SECOND ONE THIS IS A NORMAL
B6 MOUSE BUT THE TUMOR CELLS
DIDN'T EXPRESS THE RESTRICTIONAL
ELEMENT.
AS I SHOW IN THE PREVIOUS
EXPERIMENT.
THE THIRD GROUP IS THE RESULT
ENDOGENOUS T1B CELLS.
AGAIN YOU SEE THE RESPONSE.
THE FOURTH GROUP IS WHEN YOU
LOSE BOTH THE -- AND
RESTRICTIONAL ELEMENT, YOU START
SEEING -- TREATMENT BUT EVERY
CASE, IF YOU COMPARE BETWEEN THE
RADIATION CONTROL VERSUS TREATED
GROUP, YOU DO SEE SIGNIFICANT
IMPROVEMENT, THERAPEUTIC EFFECT.
SO THIS TREATMENT CAN'T HAPPEN
WITHOUT THE HELP OF THE
ENDOGENOUS B CELLS.
FOR THE NEXT EXPERIMENT, I
WANTED TO IDENTIFY THE
ENDOGENOUS -- WHEN YOU GIVE
TYRP1 T CELLS TO MICE, WHAT KIND
OF CELLS ARE BEING AFFECTED?
IS IT -- THE PUBLICATIONS, ONE
PUBLICATION SUGGESTS IT'S
MACROPHAGE, THE OTHER SUGGEST
NEUTROPHIL, AND THEN I WANTED TO
TEST IT BY MY HAND.
I STILL DON'T -- HONESTLY, I
STILL DON'T HAVE THE ANSWER, BUT
TO ANSWER THIS QUESTION, I
PERFORM THIS EXPERIMENT.
ACTUALLY STILL ONGOING BUT I'D
LIKE TO SHARE SOME RESULTS OF
THIS EXPERIMENT.
THERE WERE FOUR GROUPS.
THE FIRST TWO GROUPS RECEIVED
B16-F10 AND THE THIRD AND FOURTH
GROUP RECEIVED WITHOUT THE
RESTRICTIONAL ELEMENT.
AND THEN THE FIRST GROUP WAS THE
T-CELL WAS MOCK TRANSDUCED T
CELLS, PLUS ANTIOX 40.
THE SECOND ONE IS -- THE T CELLS
WAS THYS IS 1.1.
THIS IS RESULT FROM THE -- SO
AFTER THE TREATMENT, I FORGOT TO
MENTION THIS, AFTER TREATMENT ON
DAY FOUR, I ISOLATED CD45
POSITIVE CELLS FROM THE TUMOR,
AND THEN PERFORMED THE CITE-SEQ
ANALYSIS.
THE REASON BEING THIS IS A
POINT -- I START SEEING THE
DIFFERENCE IN THE TUMOR SIZE.
I START SEEING THE TUMOR SHRINK
AFNLGT SO I WANTED TO SEE WHAT
WAS THE FIRST THING THAT
HAPPENS.
AND THEN SO -- I AM RUNNING
OVER.
I'M SORRY.
SO THIS IS THE RESULT WITH THE
CD45 -- AS ANALYZED BY SINGLE
CELL ANALYSIS, AND THEN AS YOU
CAN SEE, THERE IS CD4 T CELLS --
AS YOU CAN SEE VERY MINOR
POPULATION WITHIN THIS CD 45
POSITIVE CELLS.
THERE ARE MACROPHAGE -- NSM K
C -- NK CELLSAND THEN NEUTROPHI.
I WANTED TO SHOW YOU THE BIG
PICTURE OF WHAT CHANGE IS
HAPPENING IN THESE DIFFERENT
CELLS AND THEN ROUGHLY SPEAKING,
I THINK YOU CAN SEE MAJOR GENE
EXPRESSION CHANGE ON THE -- WHEN
T -- GIVEN TO MICE CARRYING THE
B16-F10 WITHOUT THE
RESTRICTIONAL ELEMENT.
WHEN B16 DOES NOT HAVE THE
RESTRICTIONAL ELEMENT, YOU SEE A
STRONGER EFFECT ON THESE OTHER
CELLS.
SO IN TERMS OF -- IN THIS -- OF
DENDRITIC CELLS, THIS SEEMS TO
BE -- MUCH DIFFERENCE BETWEEN
THESE GROUPS, AND THEN IN TERMS
TERMS -- IN THE INTEREST OF
TIME, THERE IS SOME SIGN OF
INTERFERON RESPONSE, BUT NOT
MUCH DIFFERENCE.
I WANT TO SHOW YOU WHAT HAPPENED
TO THE CD8 T CELLS, DENDRITIC
CELLS IN TERMS OF THE NUMBER,
YOU SEE MORE CD8 T CELLS WHEN
TYRP1 T CELLS ARE GIVEN TO THE
RICE -- LOSS.
AND THEN ALL OF THESE CELLS,
THIS GRAPH SHOWS THE CD8 T CELLS
AND ALL THESE ARE -- ALL DERIVED
FROM -- THAT SURVIVED THE
OPERATION AND THEN PROLIFERATED
IN RESPONSE TO THE CD4 T CELLS
GIVEN TO THESE MICE.
AND THEN THIS IS SHOWING ON THE
LEFT SIDE, YOU SEE WHAT HAPPENED
TO CD8 T CELLS IN B16-F10 -- ON
THE RIGHT SIDE IS MHC LOSS --
CD8 T CELLS.
THE GREEN GENES OR -- PATHWAY
INVOLVE THE GENES.
SO IT SHOWS THAT METABOLISM OF T
CELLS HAVE CHANGED -- SHIFTED
FROM OX 4 TO -- PATHWAY, AND
THEN ALSO YOU SEE THE EFFECTOR
MOLECULE SUCH AS -- GRZ AND SO
ON, AND IN ADDITION, YOU SEE THE
GENES THAT ARE ASSOCIATED WITH
EFFECTOR DIFFERENTIATION.
YOU SEE THE DOWN REGULATION,
HERE YOU SEE THE DOWN REGULATION
OF KF2 OR -- THAT'S BACK TO AND
THEN -- SO ID2 IS HIGHLY
EXPRESSED SO THIS -- GIVING
TYRP1 T CELLS IN THE CONTEXT
OF -- RESTRICTIONAL ELEMENT
RESULTED IN THE FURTHER
DIFFERENTIATION OF ENDOGENOUS
CD8 T CELLS.
SO AT THIS POINT, THAT'S ALL I
HAVE, AND THEN IF I -- I DON'T
THINK I HAVE TIME TO SUMMARIZE
BUT -- I'M GOING TO READ THROUGH
VERY QUICKLY.
SO THE SIGNIFICANT FREQUENCY OF
LOSS OF NEOANTIGEN-SPECIFIC
CLASS I MHC ALLELE IN PATIENT
TUMORS THE HIGHER PREVALENCE OF
CD4 NEOANTIGEN REACTIVITIES IN
NON-MELANOMA EPITHELIAL CANCERS
AND SEVERAL EXAMPLES OF CD4 T
CELLS SUCCESSFULLY TREATING
TUMORS IN HUMANS HIGHLIGHT THE
GROWING UP IMPORTANCE OF CD4
T-CELL BASED THERAPIES.
AND THE CD4 T CELLS APPEAR TO
MEDIATE ANTITUMOR EFFECTS
THROUGH THREE KEY MECHANISMS.
ONE IS ACTIVATION OF HOST
MACROPHAGES AND THE NEUTROPHILS,
WHICH IS MHC INDEPENDENT AND
VERY ATTRACTIVE BUT MECHANISM
THAT WE DO NOT KNOW IF IT
TRANSLATES TO HUMAN.
AND THEN DIRECT RECOGNITION AND
ACTIVATION OF ENDOGENOUS CD8
T-CELL RESPONSES.
AND THEN THE FIRST MECHANISM
NEEDS FURTHER INVESTIGATION,
FURTHER STUDY ABOUT THE OTHER
IMMUNE COMPONENTS IN THE TUMOR
TO UNDERSTAND BETTER.
AND THEN -- SO THE LAST POINT IS
THE POINT THAT I JUST MADE, CD8
T CELLS ARE BETTER ACTIVATED
WHEN TUMOR CELLS DID NOT EXPRESS
THE PLUS 2 MHC.
OKAY.
FUTURE PLAN, I WILL SKIP THIS,
AND THEN THIS IS THE
ACKNOWLEDGMENT.
SO SORRY FOR RUNNING OVER.
I WOULD BE GLAD TO TAKE
QUESTIONS.
[APPLAUSE]
>> ONE QUICK QUESTION AND THE
THEN A QUICK COMMENT.
KEN, YOU THAT FOR EMPHASIZING
THE IMPORTANCE OF CD4 CELL,
BECAUSE I WORK ON CD4 CELL FOR A
LONG TIME AND WITH DR. ROSENBERG
AND I HAVE -- BUT I WANT TO JUST
TELL YOU, WE -- IN HUMAN AND --
AND PARTICULARLY I WANT TO THANK
NCI FOR SUPPORTING OUR
PROGRAM -- AND TODAY HERE AND I
WANT TO TALK WITH YOU MORE.
THAT IS -- TCR.
SO WORK FAIRLY WELL IN THE LUNG
CANCER, BREAST CANCER, COMPLETE
ELIMINATE CANCER CELL.
BECAUSE -- FOR FIVE YEARS, I
NEVER ABLE TO ERADICATE THE
BREAST CANCER BUT WHEN I START
WORKING ON CD4 CELL, COMPLETELY
ERADICATE CANCER CELL.
IT'S AMAZING RESULT.
THANK YOU.
>> VERY INTERESTING.
I'D LIKE TO TALK ABOUT IT.
>> OKAY.
BEFORE WE ADJOURN, I JUST HAVE A
FEW REMARKS AND SOME
INSTRUCTIONS.
I'D LIKE TO JUST SAY HOW PROUD I
AM TO NOT ONLY HAVE BEEN A
SURGERY BRANCH ALUMNI, BUT HAVE
ALSO SPENT THAT COVETED ONE-YEAR
IMMUNOTHERAPY FELLOWSHIP UNDER
DR. ROSENBERG, AND I ECHO JIM'S
COMMENTS THAT THIS IS REALLY A
LIFE-CHANGING EXPERIENCE.
AND NOW I JOINED THE VERY
TALENTED TEAM AT IOVANCE WHO HAD
A REMARKABLE 2024 WITH THE FOOD
AND DRUG ADMINISTRATION
APPROVING THE FIRST
TUMOR-ACQUIRED AUTOLOGOUS T-CELL
THERAPY FOR PATIENTS WITH
REFRACTORY METASTATIC MELANOMA.
AND REALLY THIS IS THE TIP OF
THE ICEBERG.
WE ALSO HAVE AN EXPANDING
PIPELINE FOR EARLIER INDICATIONS
AS WELL AS BROADER INDICATIONS.
SO WE'RE VERY EXCITED ABOUT THE
WORK THAT WE'VE DONE, YET THAT
HAS BEEN 50 YEARS IN THE MAKING
AND NONE OF THAT WOULD BE
POSSIBLE WITHOUT THE WORK THAT
YOU HAVE DONE HERE AS A PIONEER,
DR. ROSENBERG.
YOUR CLINICAL STAFF, MANY WHOM
ARE IN THIS ROOM, THE STAFF OF
THREE NORTHWEST AGAIN, WE'RE SO
GRATEFUL FOR THAT, WE'RE
GRATEFUL FOR WHAT YOU'VE DONE
FOR US, FOR OUR PATIENT, SOME OF
WHOM ARE ACTUALLY IN THIS ROOM
TODAY.
SO THANK YOU FOR THAT, AND
IOVANCE IS PROUD TO BE A
SUPPORTER OF TODAY'S SYMPOSIUM.
SO WE ARE TO RECONVENE AT
1:20 SHARP, I'M TOLD, SO PLEASE
BE ON TIME, BUT WE WILL NOW
ADJOURN FOR THE POSTER AND
LUNCH.
THANK YOU.
THANK YOU FOR COMING, ANOTHER
THANK YOU FOR THE SPONSOR OF
LUNCH AND POSTER SESSIONS TODAY
AS WELL AS REFRESHMENTS AT THE
BREAKS.
IT IS MY HONOR TOLL START THE
ROUNDS OF INTRODUCTIONS BY
ASKING DR. MONICA BERTAGNOLLI TO
COME TO THE STAGE.
SHE'S THE 17th DIRECTOR OF THE
NIH, SHE IS THE FIRST SURGEON,
ONLY THE SECOND WOMAN TO EVER
HOLD THAT POSITION.
[APPLAUSE]
SHE PREVIOUSLY SERVED AS 16th
DIRECTOR OF NCI, AND I GOT TO
KNOW HER WHEN SHE WAS RICHARD
WILSON PROFESSOR OF SURGERY AT
THE BRIGHAM.
WITHOUT FURTHER ADO, DR.
BERTAGNOLLI.
>> THANK YOU.
[APPLAUSE]
I GET THE BEST JOB OF THE DAY, I
GET TO INTRODUCE OUR DR. STEPHEN
ROSENBERG.
LET ME JUST SAY, STEVE HAS BEEN
ONE OF MY HEROES SINCE 1985.
THAT WAS THE YEAR I BECAME AN
INTERN IN SURGERY AT BRIGHAM AND
WOMEN'S HOSPITAL.
IN MEDICAL SCHOOL, I WANTED TO
BE AN IMMUNOLOGIST.
I LOVED IMMUNOLOGY.
BUT THE FIRST TIME I ENTERED AN
OPERATING ROOM I KNEW I HAD TO
BE A SURGEON.
IT HAPPENS TO SURGEONS.
AND I WAS ACTUALLY -- YOU KNOW,
THEN I THOUGHT, I HAVE TO MAKE A
DECISION, A CHOICE.
I'M EITHER GOING TO PURSUE
SURGERY OR BE AN IMMUNOLOGIST.
I DECIDED I WOULD DO A SURGERY
RESIDENCY.
IT JUST DIDN'T SEEM POSSIBLE TO
EXCEL IN BOTH FIELDS.
SO IMAGINE HOW THRILLED I WAS
WHEN I ENTERED MY INTERNSHIP,
FOUND OUT IT WAS NOT ONLY
POSSIBLE TO CONTINUE TO STUDY
TUMOR IMMUNOLOGY WHILE PURSUING
A CAREER IN SURGERY, IT WAS
ACTUALLY HIGHLY ENCOURAGED.
AND IN FACT, A WIDELY, WIDELY
ACCLAIMED SURGEON WHO TRAINED AT
BRIGHAM AND WOMEN'S WAS
TRANSFORMING THE FIELD OF
IMMUNOLOGY SHOWING HOW ANTITUMOR
RESPONSES COULD BE DRIVEN IN
REAL PEOPLE.
NOT JUST MICE.
USING EXOGENOUSLY ADMINISTERED
CYTOKINES.
AT BRIGHAM AND WOMEN'S, STEVE
WAS A VERY BIG DEAL.
I READ EVERY ONE OF HIS PAPERS,
HE WAS A ROLE MODEL CRITICAL TO
MY GROWTH AS A RESEARCHER AND A
SURGEON.
HE'S STILL MY HERO.
I LOVE EVERY MINUTE THAT I GET
TO SPEND ATTENDING ROSENBERG LAB
MEETINGS HERE AT NIH.
IT'S FAR TOO RARE THAT I GET TO
SPEND A WHOLE LAB MEETING, BUT
IT'S WONDERFUL.
STEVE IS ALSO A VERY BIG DEAL AT
BRIGHAM AND WOMEN'S HOSPITAL FOR
ANOTHER REASON.
HE WAS MARRIED TO ALICE
O'CONNELL.
THE ALICE O'CONNELL.
HEAD NURSE, OVERALL DIRECTOR OF
THE BRIGHAM AND WOMEN'S HOSPITAL
EMERGENCY ROOM.
AT THE BRIGHAM, ESPECIALLY IN
THOSE DAYS, EMERGENCY ROOM WAS
THE NERVE CENTER OF THE ENTIRE
HOSPITAL, AND, BOY, YOU WOULD BE
ABSOLUTELY INSANE TO MESS AROUND
WITH THE DIRECTOR.
[LAUGHTER]
SO I'M TOLD BY THOSE WHO WERE
THERE AT THE TIME THAT STEVE'S
RELATIONSHIP WITH ALICE WAS
ANOTHER THING THAT MADE HIM
INVINCIBLE.
[LAUGHTER]
AND OVER THE YEARS, AS I'VE HAD
THE PLEASURE OF KNOWING BOTH OF
THEM, IT'S CLEAR THAT THIS IS
STILL VERY TRUE TODAY.
SO, STEVE, WE ALL KNOW YOU'RE
NOT ONE TO SEEK GLORY FOR
YOURSELF AND EVENTS LIKE THIS
MAKE YOU VERY UNCONTRACTABLE.
WELL, TOO BAD, YOU'LL HAVE TO
SIT THERE AND TAKE IT, BECAUSE
WE'RE ALSO PROUD OF YOU AND SO
GRATEFUL FOR ALL YOU'VE GIVEN TO
US PERSONALLY AND TO THE ENTIRE
WORLD.
A FEW QUICK HIGHLIGHTS.
STEVE LEFT HIS RESIDENCY TO
PURSUE Ph.D. IN BIOPHYSICS AT
HARVARD.
HE THEN -- HE STUDIED CELL
MEMBRANES, STRUCTURE OF HUMAN
CELL MEMBRANES, HE THEN JOINED
THE PUBLIC HEALTH SERVICE, WAS
AN IMMUNOLOGY FELLOW AT NATIONAL
CANCER INSTITUTE, BECAME CHIEF
OF THE SURGICAL BRANCH HERE AT
NIH JULY 1, 1974, A POST HE
STILL HOLDS TODAY.
TWO HIGHLIGHTS FOR THOSE OF US
WHO CARE SO MUCH ABOUT WHAT
HAPPENS TREATING PATIENTS WITH
CANCER, FDA APPROVAL, 1992, IL-2
FOR METASTATIC RENAL CELL
CARCINOMA.
COUNTLESS OTHER MILESTONES ON
THE ROAD TO SHOWING THAT
IMMUNOTHERAPY CAN WORK FOR
PEOPLE WITH CANCER, FOR PEOPLE
WITH AUTOIMMUNE DISEASES, FOR
WHO KNOWS, THE ENTIRE FIELD IS
SO INCREDIBLY WIDE BECAUSE OF
STEVE'S WORK.
I JUST WANT TO END WITH ONE LAST
THING.
PERHAPS HIS GREATEST LEGACY, AND
I THINK HE MIGHT EVEN AGREE WITH
THIS, ARE THE 400 TRAINEES THAT
HAVE COME THROUGH THE SURGERY
BRANCH HERE AT THE NATIONAL
INSTITUTES OF HEALTH.
LEADERS THROUGHOUT THE COUNTRY
IN ACADEMIC MEDICAL CENTERS, IN
INDUSTRY, IN CLINICAL CARE.
IN HIS TALK TODAY HE WILL
DISCUSS HIS LATEST EFFORTS USING
TUMOR INFILTRATING LYMPHOCYTES
FOR PATIENTS WITH METASTATIC
EPITHELIAL CANCER.
I'M DELIGHTED AND HONORED TO ASK
DR. STEVE ROSENBERG TO COME TO
THE STAGE.
[APPLAUSE]
>> MONICA, THAT WAS A WONDERFUL
INTRODUCTION.
IT'S AN HONOR TO WORK HERE AT
THE NIH UNDER YOUR LEADERSHIP.
SO DELIGHTED THAT YOU'RE HERE.
I'M GOING TO VERY BRIEFLY
SUMMARIZE THIS AFTERNOON HOW WE
GOT TO THIS PLACE, AND WHERE
WE'RE HEADING IN THE FIELD OF
IMMUNOLOGY.
MANY PEOPLE DON'T REALIZE JUST
HOW RECENT THE DEVELOPMENT OF
CELLULAR IMMUNOLOGY WAS.
IN 1980s, ANTIBODIES WERE
DESCRIBED AND DOMINATED STUDIES
OF IMMUNOLOGY UNTIL THE 1960s.
IN 1958, IN FACT, THE "JOURNAL
OF IMMUNOLOGY" DID NOT EVEN
INCLUDE THE WORD "LYMPHOCYTE" IN
THE INDEX, RARE CELLS, UNKNOWN
FUNCTION, CIRCULATING IN LARGE
NUMBERS.
STARTING IN THE MID-1960s, ITS
IMPORTANCE BECAME KNOWN AS AN
INDICATOR OF ALLOGRAFT
REJECTION, TO PROTECTION AGAINST
TRANSFER OF MOUSE TUMORS FROM
ONE MOUSE TO ANOTHER.
YOU HAD TO USE LYMPHOCYTES,
GETTING THE FIELD GOING.
IN THE 1970s, JULY 1, 1974
WHEN I CAME TO THE NIH, CHIEF OF
THE SURGERY BRANCH, THERE WAS NO
CONVINCING EVIDENCE THAT HUMAN
LYMPHOCYTES WERE REACTIVE WITH
CANCER, THAT THERE WAS SUCH A
THING AS HUMAN CANCER ANTIGENS,
AND NO EFFECT OF IMMUNOTHERAPY
FOR CANCER IN HUMANS.
AND WE MADE SOME ATTEMPTS TO
UTILIZE THE IMMUNE SYSTEM.
AN EARLIER SPEAKER MENTIONED THE
FACT WE TRIED TO IMMUNIZE MANY
PIGS, AND USED THEIR LYMPHOCYTES
TRANSFERRED INTO PATIENTS, IT
WAS A NAIVE EVENT.
DID NOT CAUSE TUMOR RESPONSES
BUT AN INDICATION OF THE FACT WE
WERE THINKING HARD ABOUT WAYS WE
MIGHT USE THE NATURAL IMMUNE
SYSTEM THAT A PATIENT HAS TO TRY
TO TREAT THEIR CANCER.
VERY IMPORTANT TO THE FIELD WAS
THE DISCOVERY BY MORGAN RESETTI
AND GALLO IN 1976, ABOUT 7 YEARS
LATER THE DNA SEQUENCE OF THE
GENE CODING TRIAL 2 WAS
DESCRIBED BY TANAGUCI IN JAPAN.
BEGAN WORKING TO EXPRESS IL-2
AND E. COLI AT HIGH
CONCENTRATION, DETERMINE
BIOLOGICAL CHARACTERISTICS,
BEGAN TO APPLY USE OF IL-2 IN
THE HUMAN AND MOUSE THAT MIGHT
BE ABLE TO RECOGNIZE CANCER.
THAT BEGAN A SERIES OF EFFORTS,
KNOWING THAT IN FACT IT WAS
POSSIBLE TO CAUSE LYMPHOCYTES TO
EXPAND IN VITRO AND VERY LIKELY
IN VIVO, WE PERFORMED A SERIES
OF TRIALS HERE AT THE NATIONAL
INSTITUTES OF HEALTH, FIRST
USING NATURAL MAMMALIAN IL-2
THAT WE OBTAIN FROM DUPONT, MADE
LARGE AMOUNTS IN OUR OWN LAB TO
USE FOR TREATMENT OF ANIMAL
MODELS.
YOU HEARD A LITTLE BIT THIS
MORNING LACK CELLS WITH
RECOMBINANT IL-2, TREATED 16
PATIENTS WITH NATURAL IL-2, NONE
RESPONDED, ALL WITH WIDELY
METASTATIC DISEASE, NONE
RESPONDED TO LAK CELLS WITH IL-2
AND USED INCREASES DOSES OF IL-2
TO ATTEMPT TO TREAT PATIENTS.
AND SO WE TREATED 66 PATIENTS
WITH WIDELY METASTATIC CANCER,
NONE RESPONDED.
ALL WOULD GO ON TO DIE OF THEIR
CANCER.
THEY HAD VARIOUS HISTOLOGIC
TYPES OF DISEASE.
UNTIL NOVEMBER OF 1984, WE
FINALLY UTILIZED A HIGH DOSE
BOLUS IL-2, HALF-LIFE IN HUMANS,
DEVELOPED BY JIM YANG, AND IT
WAS THIS PATIENT WHO GAVE ME HER
PERMISSION TO USE THIS SLIDE,
THE FIRST PATIENT WITH
METASTATIC MELANOMA THAT
RESPONDED TO INTERLEUKIN-2
THERAPY, WIDESPREAD MEDICAL NO,
NO,
MA'AM -- MELANOMA, DISEASE FREE
TODAY 40 YEARS LATER.
WHEN YOU'RE TRYING TO DO
SOMETHING AND IT HASN'T EVER
WORKED, YOU DON'T KNOW WHETHER
WILL WORK.
ONCE YOU SEE IT HAPPENS ONCE YOU
AT LEAST KNOW IT'S POSSIBLE AND
CAN BEGIN TO WORK WITH IT AND
WORK ON IMPROVING IT.
WE WENT ON TO TREAT OVER 400
PATIENTS HERE IN THE CLINICAL
CENTER WITH HIGH DOSE IL-2.
MODEST RESPONSE RATES.
THE RESIST RESPONSE WAS 17%,
HALF OF THOSE PATIENTS HOWEVER
WERE COMPLETE RESPONDERS.
AND IF YOU HAD A COMPLETE
RESPONSE, AS YOU HEARD EARLIER
THIS MORNING, YOU WERE VERY
LIKELY TO SUSTAIN THAT COMPLETE
RESPONSE AND VERY LIKELY BE
CURED.
AND THE FDA APPROVED
INTERLEUKIN-2 FINALLY IN 1992
FOR PATIENTS WITH METASTATIC
RENAL CANCER, 1998 FOR PATIENTS
WITH METASTATIC MELANOMA.
THAT LED US TO BEGIN STUDYING
HOW DOES IL-2 MEDIATE REGRESSION
OF CANCER IN HUMANS.
AND THAT LED US TO EXAMINE A
LARGE NUMBER OF CELL TYPES
WITHIN THE HUMAN PERIPHERAL
BLOOD, LYMPH NODES, FINALLY
SETTLED ON CELL CALLED TUMOR
INFILTRATING LYMPHOCYTE, WITHIN
THE CANCER STROAMA ITSELF THESE
ARE IMMUNE CELLS THAT INFILL
INFILTRATE
INTO THE STROMA.
WHEN YOU TAKE THEM OUT THEY CAN
RECOGNIZE AUTOLOGOUS TUMOR, IN
VITRO ASSAYS, PUBLISHED IN A
MOUSE IN 1986, IN THE HUMAN IN
1987.
AND 1988 FINALLY BEGAN THE FIRST
CLINICAL TRIALS WITH TUMOR
INFILTRATING LYMPHOCYTES IN
HUMAN, PUBLISHED THIS IN THE NEW
ENGLAND JOURNAL OF MEDICINE.
THESE WERE MODEST RESPONSES.
THEY TENDED NOT TO BE VERY
DURABLE BUT DID SHOW THAT IN
FACT INTERLEUKIN-2 COULD IN
PATIENTS WITH MELANOMA MEDIATE
SOME REGRESSIONS OF SUBSTANTIAL
VOLUMES OF DISEASE.
WE TREATED 192 PATIENTS IN THE
CLINICAL CENTER WITH AUTOLOGOUS
TILL, PUBLISHED 2011, INTRODUCED
LYMPHODEPLETING CHEMOTHERAPY,
LARGELY WORKED ON BY MARK
DUDLEY, ADDED IL-2, FOLLOWING
ADMINISTRATION OF CELLS TO KEEP
THOSE CELLS ALIVE, AND FROM
THOSE MODEST RESULT ALSO THAT WE
REPORTED, IN SCIENCE IN 1986, WE
ACHIEVED OBJECTIVE RESPONSE
RATES BY RHESUS CRITERIA, 56%,
AND OF THE COMPLETE RESPONDERS
COMPRISED 25% OF ALL PATIENTS.
AND OF THE 48 PATIENTS THAT HAD
A COMPLETE RESPONSE, ONLY 2
RECEIVED MORE THAN A SINGLE
TREATMENT.
YOU HEARD FROM DR. YANG EARLIER.
THEY CAN EXPAND 10,000-FOLD IN
THE FIRST TWO WEEKS AFTER THEY
ARE ADMINISTERED, AND THEY CAN
RECOGNIZE THE CANCER AND MEDIATE
ITS DESTRUCTION.
AGAIN, IF HAD YOU A COMPLETE
RESPONSE, ONE OF THOSE 48, ONLY
2 HAD EVER RECURRED IN THE FIRST
10 YEARS FOLLOWING TREATMENT,
AND SUBSEQUENTLY.
WERE VERY LIKELY CURED OF THEIR
MELANOMA.
THE FDA, THANKS TO EXTRAORDINARY
EFFORTS GUY BIOTHERAPEUTICS WHO
TOOK THIS TO THE FDA REPEATEDLY,
FINALLY RECEIVED FDA APPROVAL IN
FEBRUARY OF 2024, JUST SEVERAL
MONTHS AGO.
IT'S NOW MORE WIDELY AVAILABLE.
SO THERE WAS A 36-YEAR INTERVAL
FROM THE FIRST DESCRIPTION OF
TIL, IN 1988, TO ITS FDA
APPROVAL IN WIDESPREAD
APPLICATION.
VERY SOON AFTER BEGINNING TO
WORK WITH CELLS, WE BEGAN TO
WONDER WHETHER OR NOT IT MIGHT
BE POSSIBLE TO GENETICALLY
MODIFY THOSE CELLS AND IMPROVE
THEIR ACTIVITY.
AND I BEGAN WORKING WITH FRENCH
ANDERSON AND MICHAEL BLAZE, HERE
AT THE NIH, TO SEE IF WE
COULDN'T GENETICALLY MODIFY
LYMPHOCYTES, INSERT THEM INTO
PATIENTS, AND LOOK FOR
IMPROVEMENTS IN THEIR ACTIVITY.
INITIAL STUDIES, HOWEVER, DEALT
WITH A PROPOSAL WE MADE TO A
VARIETY OF THE REVIEW GROUPS TO
INSERT A GENE, BACTERIAL GENE
ENCODING NEOMYCIN
PHOSPHO-TRANSFORAYS THAT WOULD
ALLOW US TO TRACK CELLS INSIDE
THE BODY AFTER THEY WERE
ADMINISTERED, DISTINGUISH THEM
FROM ENDOGENOUS CIRCULATING
LYMPHOCYTES.
THIS TURNED OUT TO BE QUITE A
COMPLEX APPROVAL PROCESS, SO I
TOOK THIS PAGE FROM MY
NOTEBOOKS, I KEPT TRACK OF THE
REVIEW GROUPS THAT WE HAD TO GO
THROUGH, AND A VARIETY OF
INSTITUTES AT THE NIH BECAUSE IT
INVOLVED SEVERAL OF US,
DIFFERENT BRANCHES.
THE RECOMBINANT DNA ADVISORY
COMMUNITY, THE RAC, PROVIDED
FINAL APPROVED BEFORE WE COULD
BEGIN.
AND THEY FINALLY HAD A VOTE IN
DECEMBER OF 1988, TO SEE IF IN
FACT THEY WOULD APPROVAL OUR
ABILITY TO DO THIS.
THE VOTE UNFORTUNATELY WAS 16 IN
FAVOR, 5 AGAINST.
JAMES WYNGAARDEN, THEN DIRECTOR
OF THE NIH, FELT BEFORE, FOR THE
FIRST TIME HUMAN FOREIGN GENES
WERE INSERTED INTO HUMANS, HE
INSISTED THE RAC APPROVE IT
UNANIMOUSLY.
WE HAD TO GO BACK THROUGH ALL
THE REVIEW GROUPS, GIVEN THE
CHANGES THAT WE HAD TO MAKE,
FINALLY THEY HAD A REVOTE AND
VOTED 13 IN FAVOR, 0 AGAINST,
AND 1 ABSTENTION.
WE WERE GIVEN PERMISSION TO
BEGIN, TO UTILIZE THESE GENE
MODIFIED CELLS, SOMETHING WE'LL
BE TALKING ABOUT IN A FEW
MINUTES.
ONLY TO HAVE JEREMY RIFKE N FILE
A LAWSUIT, THOUGHT IT WAS
IMMORAL, SETTLED SIX MONTHS
LATERN 1989 TREATED THE FIRST
PATIENT, WENT ON TO TREAT 10
PATIENTS, FIRST 5 WE REPORTED IN
NEW ENGLAND JOURNAL, CELLS
RETROVIRALLY TRANSDUCED TO
INVERT THIS NEOMYCIN PHOSPHOR
TRANSFERASE, COULD BE DONE
SAFELY, PASS QUALITY ASSURANCE,
QUALITY CONTROL TESTS, AND THAT
ENABLED US TO BEGIN WORK
ATTEMPTING TO GENETICALLY MODIFY
CELLS.
YOU'LL HEAR FROM RICK MORGAN WHO
PLAYED A VITAL CONTROL, BUT MANY
YEARS BEFORE WE COULD INSERT
GENES THAT HELPED IMPROVE THE
ABILITY OF THE CELLS TO WORK AND
YOU'LL HEAR FROM DR. MORGAN I
THINK NEXT OF OUR STUDIES THAT
ENABLED US TO PUT T-CELL
RECEPTORS THAT RECOGNIZED
MELANOCYTE ANTIGENS INTO
PATIENTS YOU A AUTOLOGOUS
LYMPHOCYTES, COULD INSERT CELLS
AFTER GROWING THEM IN IL-2 IN
PATIENTS, SIX OF THE FIRST 20
PATIENTS WITH METASTATIC
MELANOMA HAVE AN OBJECTIVE
RESPONSE, WE WERE OFF AND
RUNNING TO FIND BETTER WAYS TO
INSERT T CELL RECEPTORS AND
GENES TO IMPROVE THE ACTIVITY OF
THE CELLS.
THAT LED FINALLY, NOT FINALLY,
BUT LED EVENTUALLY TO OF THE
DEVELOPMENT OF CAR-T CELLS,
TARGETING RECEPTORS FOR CD19, A
CELL SURFACE MOLECULE EXPRESSED
ON NORMAL B LYMPHOCYTES AND MOST
B CELL MALIGNANCIES, JIM
COKENKERFER CAME TO MY LAB WITH
THE IDEA WE SHOULD APPLY TO
HUMANS, WORKING IN ANIMAL MODELS
PRECEDED EVERYTHING WE DID IN
THE HUMAN.
HE FIRST SHOWED THAT CD19
TRANSDUCED CELLS TO ELIMINATE
LYMPHOMAS IN SYNGENEIC MOUSE
MODELS AND REPORTED FIRST
INSTANCE OF HUMAN ANTI-CDCAR
MEDIATING REGRESSION OF LYMPHOMA
IN HUMANS, FIRST TEN PATIENTS WE
TREATED IN THE SURGERY BRANCH 6
EXPERIENCED COMPLETE RESPONSE,
WIDELY PICKETED UP - PICKED UP
BY INVESTIGATORS, APPROVED BY
THE FOOD AND DRUG ADMINISTRATION
IN 2017.
THIS IS A PROBLEM WE FACE IN
CANCER IN THE UNITED STATES.
STATISTICS OF THE AMERICAN
CANCER SOCIETY, OVER 600,000
DEATHS IN THE UNITED STATES EACH
YEAR FROM CANCER.
90% OF THEM RESULT FROM THE
GROWTH OF SOLID EPITHELIAL
CANCERS, ALMOST ALL OF THE
ORGANS IN THE BODY CONTAIN
DUCTS, LINING OF THOSE DUCT ARE
EPITHELIAL CELLS, MISTAKES ARE
MAKE, MUTATIONS RESULT IN MOST
CANCERS.
AND SO ABOUT SIX OR SEVEN YEARS
AGO, WE STARTED TO DEVOTE ALL OF
OUR EFFORTS TO TRYING TO FIND
WAYS TO USE THE IMMUNE SYSTEM TO
TARGET THE SOLID EPITHELIAL
CANCERS THAT RESULT IN 90% OF
ALL PATIENTS WHO DIE OF CANCER
TODAY.
THAT'S TURNED OUT TO BE QUITE A
THORNY PROBLEM.
MAJOR CHALLENGE COULD BE
FRONTING ALL OF ONCOLOGY BUT
CERTAINLY CANCER IMMUNOTHERAPY
IS DEVELOPMENT OF EFFECTIVE
IMMUNOTHERAPIES FOR THAT GROUP
OF PATIENTS THAT CANNOT BE CURED
BY ANY AVAILABLE TREATMENT.
THE CHECKPOINT MODULATORS HAVE
HAD MAJOR IMPACT ON ONCOLOGY,
BUT IN FACT SOLID EPITHELIAL
CANCERS, G.I. CANCERS, BREAST
CANCER, OTHERS, TEND NOT TO BE
RESPONSIVE TO THE CHECKPOINTS.
AND SO WE BEGAN A SERIES OF
EFFORTS TO ASK WHETHER OR NOT
ADOPTIVE CELL THERAPY COULD BE
EFFECTIVE FOR TREATMENT OF SOLID
CANCERS.
I'LL TALK ABOUT THREE APPROACHES
NOW IN PROGRESS IN THE SURGERY
BRANCH, ATTEMPTING TO USE
AUTOLOGOUS LYMPHOCYTES TO TARGET
THESE SOLID EPITHELIAL CANCERS.
WE TRY TO USE TIL.
AS WE GREW THEM FROM MELANOMAS
TO TREAT, IT DID NOT WORK.
MELANOMA, YOU HAVE TO TAKE ANY
LIMB LOW SITES -- LYMPHOCYTES,
PUT THEM IN A TUMOR, LET THEM
GROW, IF YOU ADMINISTER THEM
FOLLOWING CHEMOTHERAPY INDUCE
ANTES TUMOR RESPONSES IN OVER
HALF OF MEDICAL MELANOMA PATIE.
THAT DOES NOT WORK IN SOLID
EPITHELIAL PATIENTS, MUCH LESS
RESPONSIVE TO IMMUNOLOGIC
MANEUVERS, AND THAT MEANT WE HAD
TO SOMEHOW SEE IF THERE WERE
LYMPHOCYTES IN THESE PATIENTS
THAT WE COULD SELECT AND
SELECTIVELY GROW THEM TO SEE IF
WE COULDN'T CAUSE ANTITUMOR
RESPONSES.
AN IMPORTANT CLUE ASKING WHETHER
OR NOT -- FINDING WHAT TIL WERE
RECOGNIZING ON TUMORS WAS OUR
OBSERVATION THAT THERE WAS NO
OFF-TUMOR ON-TARGET TOXICITY IN
PATIENTS, THAT IS NORMAL CELLS
WERE NOT INTERFERED WITH, AND
THAT LED US TO EXPLORE THE ROLE
OF THE MOST SPECIFIC VARIATION
IN CANCER COMPARED TO NORMAL
CELLS, THAT IS ACCUMULATION OF
CANCER MUTATIONS, ASK WHETHER OR
NOT COULD SPECIFIC TARGETING OF
THESE ANTIGENS IF THEY WERE
IMMUNOGENIC LEAD TO TUMOR
REGRESSION.
THE ADVANTAGE OF TARGETING
MUTATIONS AS A TARGET VIRTUALLY
ALL CANSERS IS HAVE MUTATIONS IF
ONE COULD DEVELOP AND APPROACH
TARGET MUTATIONS ONE MIGHT
POTENTIALLY BE ABLE TO TREAT A
WIDE VARIETY OF DIFFERENT CANCER
TYPES.
THAT LED INVESTIGATORS IN
SURGERY BRANCH TO DEVELOP A
BLUEPRINT FOR IDENTIFYING
MUTATION REACTIVE CELLS AND
COMMON EPITHELIAL CANCERS,
RESECT TUMOR IF YOU JUST FOLLOW
THIS CLOCKWISE, WE WOULD RESECT
THE TUMOR, ISOLATE THE GENOMIC
DNA AND RNA, WHOLE EXOME
TRANSCRIPTOME SEQUENCING TO
DETERMINE WHICH GENES WERE BEING
EXPRESSED.
WE THEN IN BRIEF WOULD THEN
SYNTHESIZE 25 SURROUNDING EVERY
NON-SYNONYMOUS MUTATION IN THE
CANCER COMPARED TO NORMAL CELLS
AND USING PEPTIDES 25 PEPTIDES
WITH MUTATION IN THE MIDDLE OR
MANY GENES INSERTED INTO A
PATIENT'S ANTIGEN-PRESENTING
CELLS THAT WOULD PRESENT THOSE
ANTIGENS ON MHC MOLECULES OF
THAT PATIENT, WOULD CULTURE TIL
THAT CAUSED TUMOR REGRESSIONS IN
PATIENTS, SOME THAT DID NOT.
CO-CULTURE THAT WITH THESE
ANTIGEN-PRESENTING CELLS
EXPRESSING ALL OF MUTATIONS IN
THAT CANCER, AND BY IMMUNOLOGIC
ASSAYS INTERFERON GAMMA
SECRETION, UPREGULATION OF
ACTIVATION MOLECULES SUCH AS OX
40 COULD IDENTIFY IN POOLS
ANTIGENS THAT WERE BEING
RECOGNIZED BY TIL CELLS.
THE KEY AGAIN WAS TO TAKE THIS
25 MER WITH NON-SYNONYMOUS
MUTATION IN THE MIDDLE, T CELLS
WILL RECOGNIZE PEPTIDES 9 TO 25
AMINO ACIDS ON THE SURFACE,
PUTTING THE MUTATION IN THE
MIDDLE OF THIS CONSTRUCT
SURROUNDED BY NORMAL AMINO ACID,
EVERY POTENTIAL 9 TO 25 PEPTIDE
WOULD BE POTENTIALLY EXPRESSED
IN THIS 25 MER, AND BY THIS KIND
OF ASSAY, NO NEED TO PREDICT
PEPTIDE BINDING OF THESE
MUTATION, ALL WERE BEING TESTED
AT THE SAME TIME.
ALL CANDIDATE PEPTIDES AND MHC
LOCI, OF COURSE A PEPTIDE
PRESENTED ON MHC ON THE SURFACE
THAT IS RECOGNIZED BY IMMUNE
CELLS, AND THERE WERE NO TUMOR
CELL LINES NECESSARY.
IT'S VERY DIFFICULT TO GROW
EXCEPT FOR MELANOMA AND CERTAIN
OTHER CANCERS, DIFFICULT TO GROW
LONG-TERM LINES OF THE COMMON
EPITHELIAL CANCERS, BY STRINGING
MUTATIONS TOGETHER WE HAVE AN
AVATAR OF THE TUMOR ITSELF THAT
WE COULD STUDY.
WE THEN STUDIED MANY PATIENTS
WITH MELANOMA, BUT 130
CONSECUTIVE PATIENTS WITH
GASTROINTESTINAL CANCERS.
MEDIAN OF 123 MUTATIONS PER
CELL.
THESE 130 PATIENTS THEREFORE HAD
20,046 MUTATIONS, SCREENED
INDIVIDUALLY 15,000 OF THOSE
MUTATIONS, THEY WERE THE ONES
BEING EXPRESSED IN THE TUMOR AND
BY DOING THAT WE COULD IDENTIFY
210 IMMUNOGENIC MUTATIONS,
MUTATIONS RECOGNIZED BY A
PATIENT'S T CELL, IN THESE 130
PATIENTS.
INTERESTINGLY 1.3% OF ALL OF THE
MUTATIONS TURNED OUT TO BE
IMMUNOGENIC, AND THAT'S A NUMBER
THAT TURNED OUT TO BE THE CASE
FOR VIRTUALLY EVERY TUMOR WE
LOOKED AT, MELANOMA, BREAST
CANCERS, AND OTHERS.
INTERESTINGLY, HALF OF THE CELLS
THAT RECOGNIZE THESE TUMOR
ANTIGENS, MUTATED ANTIGENS, WERE
CD8, HALF CD4, AND THE BIG
SURPRISE IN THIS WAS THAT OF
THESE 210 NEOANTIGENS RECOGNIZED
BY A PATIENT'S T CELLS, 209 WERE
ABSOLUTELY UNIQUE ONLY TO THE
PATIENT THAT BORE THAT CANCER
AND WAS NOT SHARED BY ANY OTHER
PATIENT.
THE ONLY ANTIGEN THAT WAS SHARED
BY TWO PATIENTS WAS A KRAS G12 D
MUTATION RESTRICTED BY CW 0802,
AND THAT POINTED TO ONE OF THE
POTENTIAL PROBLEMS OF THE NEED
FOR SPECIALIZATION,
PERSONALIZATION OF TREATMENTS
BECAUSE EACH PATIENT WAS
RECOGNIZING SOMETHING DIFFERENT
IN THEIR OWN CANCER.
WE STUDIED 205 CONSECUTIVE
PATIENTS, A VARIETY OF CANCER,
GASTROINTESTINAL, BREAST, SMALL
CELL LUNG CANCER, OVARIAN,
PROSTATE, IF YOU GO DOWN THE
LIST OF ALL THE CANCERS ABOUT
3/4 OF PATIENTS WILL HAVE T
CELLS IN THEIR TUMORS, OR
CIRCULATING, THEY CAN BE
DETECTED IN THE PERIPHERAL
BLOOD, ALTHOUGH RARER FREQUENCY,
ALL OF THOSE WERE UNIQUE TO THE
INDIVIDUAL PATIENT WITH, AGAIN,
ONLY THIS KRAS G12 D MUTATION
DETECTED BY TIL SHARED BY
PATIENTS, 363 NEOANTIGENS
RECOGNIZED.
THAT THEN LED TO REALIZATION OF
WHAT A CANCER ANTIGEN WAS,
BASICALLY ANY INTRACELLULAR
PROTEIN, APPEARED TO BE RANDOM
SOMATIC MUTATIONS, COULD
POTENTIALLY BE A CANCER ANTIGEN
IF IT WAS MUTATED, PROCESSED
EXTRACELLULARLY TO A PEPTIDE.
COMPLEX MOLECULE, 1 IN 70 WERE
NEOANTIGENS, RECOGNIZED BY T
CELLS.
SINCE ALMOST ALL PATIENTS HAVE
MUTATIONS IF WE COULD FIGURE OUT
A WAY TO TARGET THEM PATIENTS OF
MANY CANCER TYPES WOULD BE
ELIGIBLE.
BAD NEWS, TREATMENT IS GOING TO
HAVE TO BE HIGHLY
INDIVIDUALIZED.
AND THEREFORE COMPLEX.
BUT, AGAIN, IF WE COULD FIND
THINGS THAT WORK, WE HAD EVERY
CONFIDENCE THAT THE GENIUS OF
AMERICAN INDUSTRY WOULD FIGURE
OUT WAYS TO MAKE IT AVAILABLE.
WELL, THAT BEGAN THEN A CLINICAL
TRIAL.
YOU'VE SEEN A VARIATION OF THIS
SLIDE BEFORE, AGAIN IF YOU
FOLLOW EXCISION OF TUMOR
CLOCKWISE, WE NOW EXCISE 1 OR
MORE FROM AN INDIVIDUAL CANCER
PATIENTS, ALL OF THE PATIENTS
THAT WE TREAT HAVE ADVANCED
METASTATIC MALIGNANCY THAT HAVE
BEEN THROUGH ALL STANDARD
TREATMENTS, AND OF COURSE FDA
ONLY ALLOWS EXPERIMENTAL
TREATMENTS OF UNKNOWN
EFFECTIVENESS AND UNKNOWN
TOXICITIES, IN THAT DIFFICULT
PATIENT POPULATION.
WE PLATE 24 TO 48 FRAGMENTS FROM
THE TUMOR, GOOD SPATIAL
RECOGNITION OF LYMPHOCYTES FROM
A VARIETY OF SITES.
EVERY INDIVIDUAL FRAGMENT IS
TESTED FOR REACTIVITY, AGAINST
THE PATIENT'S TIL CELLS, THAT
ENABLES US THEN TO FIND TIL THAT
LETS US IDENTIFY THE ANTIGEN.
THOSE FRAGMENTS SHOWN IN VITRO
TO RECOGNIZE A FRAGMENT WE THEN
SELECT THAT FRAGMENT OR
FRAGMENTS ONLY AND GROW THEM IN
VITRO IN OUR GMP FACILITY, AND
THEN INFUSE THEM FOLLOWING A
LYMPHODEPLETION OF THE PATIENT
THAT HAS MANY ADVANTAGES,
PREDOMINANT BEING ELIMINATION OF
T REGULATORY CELLS, MYELOID
DERIVED SUPPRESSORS, STIMULATES
ENDOGENOUS CYTOKINE PRODUCTION.
AND THE FIRST PATIENT WE DID
THIS ON HAPPENS TO BE IN THE
AUDIENCE.
I WASN'T AWARE SHE WAS GOING TO
BE HERE, 45-YEAR-OLD WOMAN WHO
HAD A METASTATIC CARCINOMA, 45
YEARS OLD, SIX CHILDREN.
HAD A RIGHT HEPATECTOMY,
DEVELOPED LUNG AND LIVER
METASTASES, RECEIVED MULTIPLE
CHEMOTHERAPY REGIMENS, DID NOT
RESPOND.
WE GAVE HER UNSELECTED TIL
GROWING ALL THE CELLS AND GIVING
THEM, SHE DID NOT RESPOND.
WE THEN SELECTED A FRAGMENT THAT
SEEMED TO BE MUCH MORE REACTIVE
THAN OTHERS.
SHE HAD 26 MUTATIONS.
ERIC TRAN IN THE LAB IDENTIFIED
CELLS THAT RECOGNIZED HER B1 IB
CANCER MUTATION.
ON YOUR LEFT IS AN EXAMPLE OF
THE LARGE TUMORS THAT THIS
PATIENT HAD GROWING IN HER
LUNGS, CAN YOU SEE YELLOW ARROWS
POINTING TO THEM.
LIVER METASTASES AS WELL.
SHE UNDERWENT DRAMATIC
REGRESSION.
ABOUT 3 1/2 YEARS LATER.
HOWEVER, A FEW NODULES APPEARED
IN RIGHT LUNG, RESECTED, SHE
BECAME AND REMAINS DISEASE FREE
TO THE PRESENT TIME.
MELINDA, IT'S A DELIGHT TO HAVE
YOU HERE.
PLEASE COME AND SAY HELLO TO ME.
[APPLAUSE]
WE BEGAN A SERIES OF THREE
CONSECUTIVE TRIALS, IN PATIENTS
WITH SOLID METASTATIC EPITHELIAL
CANCERS, ALL OF WHOM HAD BEEN
THROUGH STANDARD TREATMENT.
AND THOSE THREE TRIALS THAT I'LL
DISCUSS IN THE NEXT FEW MINUTES
INVOLVE GIVING TIL NOT SELECTED,
WE TREATED 18 PATIENTS, MANY
MORE IN FACT IN TRIALS PRIOR
WITH NO RESPONSES.
WE THEN USED OUR SELECTION
CRITERIA IN 39 PATIENTS, ONLY 3
OF THOSE PATIENTS RESPONDED.
BUT THE CELLS THAT WE GAVE WERE
EXPRESSING VERY HIGH LEVELS OF
PD-1, AND EXPERIMENTS IN THE
LABORATORY SHOWED US THAT
FOLLOWING THE INFUSION OF THESE
CELLS, THE FREQUENCY OF PD-1 AND
INTENSITY OF PD-1 EXPRESSION WAS
INCREASED AS CELLS GREW AND
RECOGNIZED THE CANCER IN THE
PATIENT.
THAT LED US TO A CURRENT TRIAL
NOW ONGOING, BRINGING YOU UP TO
DATE, THAT WE GIVE THESE
SELECTED TILs, ALONG WITH
PEMBROLIZUMAB, AN ANTI-PD-1
REAGENT, EIGHT PATIENTS OF 34
HAD OBJECTIVE RESPONSES BY
RHESUS CRITERIA, 23.5%.
IN MANY WAYS I FEEL THE WAY I
FELT, I FEEL NOW THE WAY I FELT
WITH THE FIRST IL-2 RESPONSES,
REALIZING IT WAS POSSIBLE TO SEE
THESE EPITHELIAL CANCERS
DISAPPEAR, STIMULATES US TO NOW
TAKE ADVANTAGE OF THAT
INFORMATION AND TRY TO IMPROVE
UPON I YOU CAN SEE EACH LINE
REPRESENTING A PATIENT, IN THIS
SPIDER DIAGRAM OF THE SIZE OF
THEIR TUMORS.
YOU CAN SEE IN THE UNSELECTED
TIL A COUPLE PATIENTS SHOWED
MINOR REGRESSIONS BUT NEVER
REACHED RECIST CRITERIA.
WHEN WE GAVE COLLECTED TIL THREE
PATIENTS GOOD SHOW A REGRESSION.
WHEN WE ADDED PEMBROLIZUMAB YOU
CAN SEE THIS MASS OF PATIENTS
WHOSE TUMORS WERE DECREASING
FOLLOWING THE ADMINISTRATION OF
THE CELLS, AND 23.5% OF PATIENTS
NOW SHOW OBJECTIVE RECIST
REGRESSIONS, AND WE'VE ONLY
SUMMARIZED NOW RECENTLY AND
SUBMITTED MANUSCRIPTS,
GASTROINTESTINAL PATIENTS WITH
METASTATIC DISEASE, THREE
PANCREATIC PATIENTS, ONE
RESPONDED, ONE
CHOLANGIOCARCINOMA, 6 OF 28
COLORECTAL, AND 24% OBJECTIVE
RESPONSE RATE, AND WE SAW TUMORS
REGRESS FROM MULTIPLE ORGANS.
THESE LARGE LUNG METASTASES
DISAPPEARED COMPLETELY IN THIS
PATIENT.
THIS PATIENT WITH A
CHOLANGIOCARCINOMA HAD BONY
METASTATIC DISEASE, ERODING
BONE, YOU CAN SEE WHEN THE
LEASES DISAPPEARED THE BONE
REFORMED.
MULTIPLE INTRAABDOMINAL LYMPH
NODES SHRANK SUBSTANTIALLY AS
WELL.
THIS PATIENT WITH COLON CANCER,
AN MRI SCAN, YOU CAN SEE
MULTIPLE LIVER METASTASES, THESE
PATIENTS PROGRESSED, SHOWN
PROGRESSION PRIOR TO ALL OTHER
TREATMENTS, DRAMATIC REGRESSION,
PARTIAL REGRESSION, OF MANY OF
THESE LESIONS.
ANOTHER PATIENT WITH MULTIPLE
LUNG DEPOSITS.
THERE'S NO ORGAN THAT HAS NOT
RESPONDED, ALTHOUGH WE DO NOT
TREAT PATIENTS WITH BRAIN
METASTASES SO I CAN'T COMMENT ON
THAT.
THIS IS A PANCREATIC CANCER
PATIENT WHO HAD TWO LESIONS, ONE
WON'T TO DISAPPEAR, THE OTHER
SHRANK.
SO PANCREATIC CANCERS ARE NOT
RESISTANT TO THIS AS WELL.
RECENTLY COMPLETE AND SUBMITTED
FOR PUBLICATION DETAILED
ANALYSIS OF THESE PATIENTS.
CONDUCTED BY FRANK LOWERY AND
STEPHANIE GOFF, AND FOUND
FACTORS ASSOCIATED WITH
OBJECTIVES RESPONSES,
UNFORTUNATELY NOT AS MANY AS WE
WISHED, BUT AS MIGHT BE
OBVIOUSLY UNDERSTOOD THE
RESPONDERS TENDED TO REACT WITH
MORE ANTIGENS, YOU SAW A LITTLE
OF THIS FROM PAUL ROBBINS THIS
MORE THAN, THAN NON-RESPONDERS.
NUMBER OF REACTIVE CD4 CELLS
RATHER THAN CD8 CELLS HIGHLY
CORRELATED WITH LIKELIHOOD OF A
PATIENT RESPONSE AND WE'RE
CONTINUING IN DETAIL THOSE
STUDIES AS WE PROGRESS.
THIS IS NOT ONLY
GASTROINTESTINAL CANCER,
ASTONISHING TO ME THIS PATIENT
IS ALSO IN THE AUDIENCE, I
DIDN'T REALIZE SHE WOULD BE
COMING BUT SHE HAD A METASTATIC
BREAST CANCER THAT HAD BEEN
THROUGH SEVEN PRIOR DIFFERENT
TREATMENTS, CHEMOTHERAPIES,
HORMONAL AND TARGETED
TREATMENTS, PRESENTED TO US WITH
MULTIPLE METASTASES IN HER
LIVER.
CAN YOU SEE IN THE CAT SCAN
PROBABLY INTERNAL MAMMARY NODE
BEGINNING TO GROW THROUGH THE
CHEST, GROW THROUGH THE CHEST
WALL.
WE TREATED HER WITH HER OWN
AUTOLOGOUS CELLS ALONG WITH
PEMBROLIZUMAB, AND EVERYTHING
HAS DISAPPEARED, AND SHE REMAINS
DISEASE FREE NOW, OVER FIVE
YEARS LATER.
LINDA, DELIGHTED THAT YOU'RE
HERE TO HELP CELEBRATE THIS
OCCASION.
[APPLAUSE]
GIVEN OUR START, WITH THE
INSERTION OF RECEPTORS, INTO A
PATIENT'S NORMAL LYMPHOCYTES TO
CONVERT NORMAL LYMPHOCYTES TO
CELLS WITH ANTITUMOR ACTIVITY WE
BEGAN A SERIES OF TRIALS IN
WHICH WE COULD IDENTIFY THE
INDIVIDUAL T-CELL RECEPTORS
RECOGNIZING PATIENTS' UNIQUE
ANTIGENS BECAUSE WE COULD
IDENTIFY THE CELL, WITH QUITE
STRAIGHTFORWARD AS YOU'LL HEAR
FROM MARIA PARKER WHO RAN THAT
STUDY, ONE COULD QUITE READILY
OBTAIN THE SEQUENCE OF T-CELL
RECEPTORS FROM THE CELLS OR GENE
SIGNATURES BEING DEVELOPED IN
THE LABORATORY BY FRANK LOWERY.
INSERT THOSE INDIVIDUAL T-CELL
RECEPTORS TARGETING UNIQUE
ANTIGENS, INTO A PATIENT, WE
REPORTED THIS IN NATURE MEDICINE
JUST A FEW MONTHS AGO.
THREE OF THE SEVEN PATIENTS THAT
WE TREATED WITH THEIR OWN UNIQUE
CELLS SHOWED OBJECTIVE
REGRESSION OF TUMOR.
AGAIN, EACH WITH DIFFERENT
ANTIGEN.
INTERESTINGLY OF THE CD8 CELLS
THAT WERE MEDIATING THESE
RESPONSE THEY HAD THE KINDS OF
CRITERIA THAT ANOTHER FELLOW IN
THE LAB IDENTIFIED AS OPTIMAL
PHENOTYPE OF MELANOMA CELLS THAT
CAUSE REGRESSION, AND THAT IS
MORE STEM-LIKE PHENOTYPE CD39
NEGATIVE CD69 NEGATIVE, LATER ON
TODAY WE'LL HEAR FROM MORIAH
PARKER ABOUT THIS TRIAL IN SOME
DETAIL.
THESE WERE UNIQUE ANTIGENS.
THERE ARE SURPRISINGLY FEW
SHARED ANTIGENS, AS YOU HEARD
FROM THE ELEGANT STUDIES THAT
YOU HEARD ABOUT THIS MORNING
FROM DR. YANG.
TWO FELLOWS IN THE LABORATORY
KIM AND LEVIN, USED A HIGHLY
TARGETED SEQUENCING APPROACH AS
WELL AS IN VITRO SENSITIZATIONS
TO TRY TO RAISE T-CELL RECEPTORS
THAT RECOGNIZED THE TWO MOST
COMMONLY EXPRESSED MUTATIONS IN
HUMANS WITH CANCER, AND THAT WAS
THE KRAS AND P53 MUTATIONS.
KRAS PRESENT IN 35%, THEY
IDENTIFIED A LIBRARY OF 14 KRAS
AND 14 P53 MUTATIONS, T CELLS,
THAT COULD RECOGNIZE THEM AND WE
PUBLISHED THAT A COUPLE YEARS
AGO, ALONG WITH THE SEQUENCE OF
ALL OF THESE RECEPTORS SO THEY
COULD BE WIDELY USED BY OTHERS.
AND SO WE BEGAN OURSELVES TO SEE
IF WE COULDN'T UTILIZE THESE
RECEPTORS TO TARGET P53 AND
KRAS, WE HAVEN'T PUBLISHED ANY
OF THIS YET.
INTERESTINGLY, WE HAVE SEEN
RESPONSES TO A PATIENT'S OWN
NORMAL CELLS, TRANSDUCED WITH
ONE OF THE T-CELL RECEPTORS IN
THIS KRAS LIBRARY.
YOU CAN SEE THESE METASTATIC
DEPOSITS IN THE LIVER.
WHICH BY THREE MONTHS HAD
DISAPPEARED.
AND THIS PATIENT WHO HAD THIS
LARGE LESION AGAIN THE
PANCREATIC CANCER PATIENT WHO
RECEIVED ALLOGENEIC CELLS
TARGETING KRAS WHICH IS
SHRINKING AND NOW STILL
SHRINKING NINE MONTHS LATER.
WE JUST SAW THIS PATIENT IN
CLINIC EARLIER THIS WEEK.
THIS PATIENT WITH A DEVASTATING
RECTAL CANCER HUGE LESIONS IN
THE LIVER, SHOWED ELIMINATION
WHEN THEY RECEIVED P53 T-CELL
RECEPTOR, AGAIN THESE WERE
RECEPTORS RECOGNIZED, THESE BY
RESEARCH BY FELLOWS IN THE
LABORATORY.
AND YOU CAN SEE THE DRAMATIC
REDUCTION OR ELIMINATION OF
SEVERAL OF THESE RECTAL CANCER
LESIONS OUT TO SEVEN MONTHS.
WELL, AS I MENTIONED, I FEEL
WE'RE AT THE START NOW OF BEING
ABLE TO TARGET THE SOLID
EPITHELIAL CANCERS WITH TIL.
THERE ARE NONE NOW THAT CAN
TARGET THIS POPULATION.
LIST HERE A SERIES OF APPROACHES
NOW BEING STUDIED IN THE BRANCH.
WE NOW NOW ADOPTIVE CELL THERAPY
CAN MEDIATION REGRESSION OF
SOLID EPITHELIAL CANSERS IS,
EFFORTS UNDERWAY TO APPROVE BY
ADDING A VACCINE THAT TARGETS
THE SAME ANTIGEN AS T CELLS
RECOGNIZE, GENE-GENE SIGNATURES
TO IDENTIFY ANTITUMOR CELLS, I
THINK THERE'S BEEN VERY
IMPRESSIVE WORK DONE BY LEVIN
AND KIM SHOWING IN VITRO
SENSITIZATION TO ENHANCE ANTIGEN
REACTIVITY AND MANY OTHERS, HAVE
EVERY EXPECTATION OR ARE WORKING
LARD TO SEE IF WE CAN'T FIND
IMPROVEMENTS.
I'D LIKE TO FINISH BY
EMPHASIZING THE FACT THAT
VIRTUALLY EVERYTHING THAT I'VE
SHOWN YOU TODAY WAS PERFORMED BY
FELLOWS IN THE LABORATORY,
EITHER SURGICAL FELLOWS THAT
CAME TO SPEND THREE YEARS WITH
US, TO LEARN TO DO RESEARCH OR
PhDs WHO CAME TO THE
LABORATORY AND WHEN THEY LEAVE
WE PUT THEIR PHOTOGRAPH UP ON
THE OUTSIDE OF THE WALL OF THE
MAIN OFFICE OF THE SURGERY
BRANCH, AND IT GOES WAY DOWN THE
HALL.
AND SO WE'RE INDEBTED TO THESE
FELLOWS, FOR ALL THE HARD WORK
THEY HAVE DONE.
WE'VE ALSO HAD AN OUTSTANDING
NUMBER OF SENIOR STAFF THAT HAVE
WORKED WITH US OVER THE YEARS,
JIM YANG CAME IN MY LAB OUT OF
HIS RESIDENCY, STAYED 40 YEARS,
HE'S BEEN A VITAL PART OF
EVERYTHING THAT WE'VE DONE, ALL
OF THE DIFFERENT APPROACHES,
PAUL ROBBINS, MOLECULAR
BIOLOGIST, MARIA PARKHURST
RESPONSIBLE FOR CELL GROWTH AND
INDIVIDUAL TCR PROTOCOLS, NICK
RESTIFO SPENT 30 YEARS IN THE
SURGERY BRANCH STUDY ANIMAL
MODELS AND DEVELOPING PRINCIPLES
UPON WHICH WE BASE OUR CLINICAL
TRIALS.
YOU HEARD FROM KENICHI, 27
YEARS, SUZANNE WAS IT'S FIRST
PERSON EVER TO GIVE A TIL TO
PATIENTS.
MARK DUDLEY RAN THE LAB THAT
GREW THE CELLS THAT WE UTILIZED
IN OUR CLINICAL TRIALS.
UDAI IS WORKING ON APPLICATION
OF TIL FOR PATIENTS, 15 YEARS,
STEPHANIE GOFF NOW IN CHARGE OF
OUR GMP FACILITIESES AND
PROTOCOLS, CAME BACK ON OUR
SENIOR STAFF AND PLAYS A VITAL
ROLE.
PATRICK HWU, REMARKABLE
SCIENTIST, FIRST PERSON TO EVER
USE CAR-T CELLS IN PATIENTS,
FIRST PERSON IN OUR LAB TO
INSERT FOREIGN GENES INTO CELLS.
RICK MORGAN WILL FOLLOW MY TALK,
TALK BECOME STUDIES INSERTING
GENES.
MIKE LOTZE, JIM MULE YOU HERDS
FROM.
I MENTION MICHELLE GAYE, STARTED
AS A SECRETARY/ADMINISTRATIVE
ASSISTANT, NOW THE GLUE THAT
HOLDS THE SURGERY BRANCH
TOGETHER, WORKED WITH ME FOR 36
YEARS, THREATENING TO RETIRE IN
DECEMBER.
[LAUGHTER]
WHICH IS A FRIGHTENING PROSPECT.
FINALLY, MY LAST SLIDE, THE
FIRST WEEK THAT I WAS AN INTERN
IN SURGERY AT THE HOSPITAL IN
BOSTON I I MET ALICE O'CONNOR,A
NURSE IN THE SURGERY WARD.
SHE MIGHT TELL THE STORY
DIFFERENTLY BUT IN FACT CHASED
ME SHAMELESSLY FOR FIVE YEARS.
[LAUGHTER]
BEFORE WE WERE FINALLY MARRIED,
AND THIS PAST YEAR WE CELEBRATED
OUR 57th WEDDING ANNIVERSARY.
NOW, I'VE WORKED HARD OVER THE
YEARS.
[APPLAUSE]
I WORKED HARD OVER THE YEARS,
AND THAT STEALS A LOT OF TIME
AWAY FROM FAMILY.
BUT ALICE HAS BEEN A REMARKABLE
SUPPORT, NEVER A WORD OF
COMPLAINT.
AND HAS BEEN A VITAL PERSON
RESPONSIBLE FOR MUCH OF WHAT
I'VE BEEN ABLE TO DO.
FIRST 40 YEARS THAT I WAS HERE,
AT THE NIH, THERE PROBABLY
WEREN'T 40 DAYS WHEN I WASN'T
TRAVELING THAT I WAS NOT IN THIS
HOSPITAL, EITHER SEEING PATIENTS
OR MEETING WITH FELLOWS ABOUT
THEIR RESEARCH.
SO, ALICE, THANK YOU FOR ALL
YOU'VE DONE.
[APPLAUSE]
COULD TAKE QUESTIONS.
[LAUGHTER]
[APPLAUSE]
>> YOU SAID THERE WERE 40 DAYS
IN THE YEARS YOU HAVEN'T BEEN AT
THE HOSPITAL.
WHERE WERE YOU?
[LAUGHTER]
>> WORD OF ADVICE FOR YOUNG UP
AND COMING PEOPLE, NEVER
MODERATE THE SESSION YOUR BOSS
IS TALKING IN.
I'D LIKE NOW TO ASK RICK MORGAN
TO COME UP, VITAL TO OUR EFFORTS
GIVING GENE THERAPY PATIENTS.
>> THANK YOU, STEPHANIE.
SO WHEN I SAW THE SCHEDULE AND
HAVE TO FOLLOW THE NIH DIRECTOR
AND DR. ROSENBERG, LIKE, OKAY,
BUT I'M A GLASS HALF FULL KIND
OF GUY.
THANK YOU FOR STAYING TO HEAR MY
TALK.
WHEN I ARRIVED HERE IN LATE
'80s AS A YOUNG POSTDOC, THIS
PLACE WAS LARGE AND
INTIMIDATING.
NOW WHEN I COME BACK IT FEELS
LIKE HOME.
SO, WHAT I'M GOING TO DO IS TELL
YOU SOME STORIES, MANY OF WHICH
ARE FROM OUR HOME HERE AT THE
NCI.
AND SET UP THE NEXT FEW TALKS SO
LET'S GO THROUGH THE ORIGINS OF
T-CELL RECEPTOR GENE THERAPY.
YOU NOW KNOW THIS CONCEPT.
IF YOU CAN IDENTIFY ANTIGEN
REACTIVE CELL, TECHNOLOGY EXISTS
TO IDENTIFY THOSE RECEPTORS THAT
ARE RECOGNIZING THE TUMOR.
SO ALPHA CHAIN, BETA CHAIN,
HETERODIMERE T-CELL RECEPTOR,
ONCE IDENTIFIED YOU CAN MOVE
THROUGH MOLECULAR CLONING
TECHNIQUES INTO A VECTOR THAT
ALLOWS YOU TO TAKE THE MOLECULE
AND GET IT INTO ANOTHER CELL.
YOU NEED TO VECTORIZE.
AND THEN YOU HAVE TO PRODUCE
THESE REAGENTS UNDER CONDITIONS
THAT YOU CAN GIVE IT TO A
PATIENT.
GOING THROUGH THE SAFETY
QUALIFICATIONS YOU NEED TO DO
BEFORE YOU CAN GIVE THIS REAGENT
TO PATIENT CELLS.
ONCE YOU'VE DONE THAT YOU VERIFY
THE POTENCY AND SPECIFICITY, AND
WHAT YOU'LL SEE AGAIN THIS IS
NOT -- IT'S A JOURNEY.
WE'VE HEARD ABOUT JOURNEYS.
T-CELL RECEPTOR GENE THERAPY IS
NO SHORTER A JOURNEY THAN THE
OTHERS YOU HEARD ABOUT TODAY.
BEGINNING IN 1986, THE VERY
FIRST MURINE T-CELL RECEPTORS
MOVED FROM ONE T CELL TO ANOTHER
T CELL.
AND THAT AFFORDED NEW T CELL
WITH THE SPECIFICITY FROM THE
ORIGINAL CELL, AGAIN SETTING THE
PREMISE YOU COULD TAKE THIS
HETERODIMER AND GET SPECIFICITY
OF ORIGINAL CELL.
OVER THE NEXT DECADE OR SO MORE
OF THESE ANTIGEN RECEPTORS WERE
CLONED PARTICULARLY IN SURGERY
BRANCH, THEY WERE THEN
TRANSFERRED IN 1995 BY MIKE'S
GROUP TO ANOTHER T CELL, SHOWED
T CELL NOW HAD ABILITY TO
RECOGNIZE THE TUMOR.
SO THAT WAS THE BASIC
TECHNOLOGY.
IT TOOK SOME TIME THEN ABOUT A
DECADE TO TAKE THAT GENE
TRANSFER TECHNOLOGY INTO ARE
EXPAND TO BE USED FOR HUMAN
CLINICAL APPLICATIONS.
SO WHAT I'M GOING TO DO IS GO
THROUGH THE APPLICATIONS, IT'S
BEEN AN AMAZING 20, 30 YEARS OF
HUMAN T-CELL RECEPTOR GENE
TRANSFER EXPERIMENTS.
FIRST TIME LYMPHOCYTES SHOWED
SHOWN IN HUMANS, A REPORT IN
"SCIENCE" IN 26.
THIS WAS TAKING A MELANOCYTE
DIFFERENTIATION ANTIGEN T-CELL
RECEPTOR CLONED BY THE BRANCH,
PUTTING INTO RETROVIRAL VECTOR,
PROTOCOL STARTS WITH APHERESIS,
YOU THEN USE GENE TRANSFER
TECHNOLOGY TO PUT IN THE NEW
RECEPTOR INTO THE T CELL, T CELL
HAS TWO RECEPTORS, THERE ARE
TRICKS TO MINIMIZE ANY POTENTIAL
CROSS-REACTIVITY BUT THAT'S A
TOPIC WHICH WE WON'T GET TO
TODAY.
EXPAND THOSE CELLS TO LARGE
NUMBERS.
RETURN THEM TO THE PATIENT FROM
WHICH THEY CAME.
NOW YOU'VE GIVEN THAT PATIENT AN
EXTRA BOOST OF A TUMOR-REACTIVE
POTENTIALLY TUMOR REACTIVE
LYMPHOCYTES, THE SAME
CHEMOTHERAPY THAT TIL PATIENTS
GET, WE DID PEPTIDE VACCINES AS
WELL.
WE DID IN A SMALL NUMBER OF
PATIENTS, ABOUT 12% OF INITIAL
PATIENTS RESPONDED, YOU COULD
GET DRAMATIC TUMOR REGRESSIONS,
LOOKING AT THE DRAMATIC MELANOMA
METASTASES TO THE DELIVER,
REGRESSION SMALL ENOUGH SURGEONS
EXCISED IT.
YOU COULD GENETICALLY ENGINEER A
NORMAL LYMPHOCYTE TO MEDIATE
TUMOR REGRESSION.
THAT ALLOWED US TO EXPAND
THINKING.
THIS INITIAL STUDY, THIS WASN'T
ACTIVE T-CELL RECEPTOR, BRANCH
FOCUSED ON MORE POTENT
RECEPTORS, YOU COULD DETECT
LOWER AMOUNTS OF POTENTIALLY
ANTIGEN ON THE SURFACE.
HERE IS AN EXAMPLE REPORTED IN
2009, ISOLATED THEM, TREATED A
NUMBER OF PATIENTS, AND INDEED
RESPONSE RATE PARTICULARLY WITH
THE MART RECEPTOR WENT UP FROM
12 TO 13% TO 13%, SHOWING
OBJECTIVE RESPONSES.
INCLUDING BRAIN METASTASES,
HAVING THAT DIFFERENTIAL ANTIGEN
AFFORDED ABILITY TO RECOGNIZE
NORMAL MELANOCYTES.
WE DID SEE TOXICITY IN THE SKIN,
EYE, IN THE EAR, WHERE NORMAL
MELANOCYTES RESIDE.
THE FIELD THOUGHT DIFFERENTLY,
ARE THERE OTHER ANTIGENS COULD
YOU TARGET WHICH DON'T HAVE THIS
ON-TARGET OFF-TUMOR TOXICITY.
ONE THOUGHT WAS CLASS OF
ANTIGENS, CANCER GERMLINE WHICH
PAUL ROBBINS REFERRED TO THIS
MORE THAN, PAUL PUBLISHED
BEAUTIFUL STUDIES SHOWING
DRAMATIC RESPONSES, DURABLE.
NOT PARTICULARLY HIGHLY
EXPRESSED, ON MOST EPITHELIAL
CANCERS.
ANOTHER CANCER TEST ANTIGEN,
MAGE, MORE HIGHLY EXPRESSED.
LOOKING AT T-CELL RECEPTORS.
IN 2013, BOTH SURGERY BRANCH AS
WELL AS CARL JUNE'S GROUP AT
UPenn PUBLISHED REPORTS WHERE
THEY WERE ISOLATED, MODIFIED,
ENHANCED REACTIVITY AGAINST
MAG-A3, GIVEN TO A NUMBER OF
CANCER PATIENTS.
IN THE SURGERY BRANCH TRIAL FIVE
OUT OF NINE PATIENTS WHO GOT
THIS ANTI-MAGE-A-3 RECEPTORS HAD
TUMOR REGRESSION.
AT THE SAME TIME, THREE OF NINE
DEVELOPED NEUROTOXICITY,
DEVELOPED WITHIN FIRST WEEK,
PROGRESSIVE, TWO DIED FROM
MASSIVE NEUROINFLAMMATORY
DISEASE.
UNRELATED BUT AT THE SAME TIME
PENN GROUP TREATED WHO PATIENTS
WITH THEIR ANTI-MAGE-A3
RECEPTORS.
BOTH GROUPS, WE DIDN'T SEE THIS
COMING.
WHAT WE FOUND WAS TWO TYPES OF
EXAMPLES.
IN THE SURGERY BRANCH EXAMPLE WE
KNEW THAT OUR MAGE A-3 RECEPTOR
COULD RECOGNIZE OTHER MEMBERS OF
THE FAMILY, 9 AND 12.
THERE'S A VERY SMALL AMOUNT,
RARE NEURONS IN THE BRAIN WHICH
EXPRESS PROBABLY MAGE-A12, WE
BELIEVE THAT'S WHAT OCCURRED IN
THE SURGERY BRANCH CASES, ENOUGH
T CELLS MADE THEIR WAY INTO CNS,
BUT NOT A UNIFORM FINDING
BECAUSE ONLY THREE OF THE NINE
PATIENTS TREATED HAD THIS
NEUROLOGICAL TOXICITY.
IN THE CASE OF THE PENN GROUP,
DIFFERENT RECEPTOR, THEIR
RECEPTOR HAD NOW AGAIN MODIFIED
RECEPTOR, ABILITY TO RECOGNIZE
DISSIMILAR PROTEIN, TITAN,
HIGHLY EXPRESSED IN
CARDIOMYOCYTES, A PEPTIDE COULD
BE RECOGNIZED BY THE T-CELL
RECEPTOR AGAINST THEIR T-CELL
RECEPTOR.
THAT WAS EXAMPLE AGAIN OF OFF
TARGET OFF TUMOR REACTIVITY.
THIS GAVE PAUSE TO THE FIELD.
WHAT ELSE CAN WE DO?
ONE CLEAR EXAMPLE OF WHAT MIGHT
BE AN IDEAL TARGET WOULD BE
VIRAL ASSOCIATED ANTIGENS FROM
FOR EXAMPLE HPV, AND CHRISTIAN
HENDRICKS IN THE BRANCH
DEVELOPED SPECIFIC RECEPTORS
AGAINST HPV A-6 PROTEIN.
IN 12 PATIENTS ADMINISTERED
THESE T CELLS TO THESE PATIENTS,
TWO PATIENTS RESPONDED.
WHAT I THINK IS REMARKABLE WAS
THEY WERE GIVEN VERY LARGE
NUMBERS OF T-CELL RECEPTOR
ENGINEERED CELLS, OVER 100
BILLION, THERE WAS NO
SIGNIFICANT TOXICITIES NOTICED.
SO IF YOU HAVE THE RIGHT T-CELL
RECEPTOR YOU CAN ADMINISTER IT,
IN LARGE NUMBERS, YOU'LL NOT SEE
POTENTIALLY TOXICITIES RELATED
SPECIFICALLY TO THE RECEPTOR PER
SE.
WHAT WAS ALSO FOUND IN
CHRISTIAN'S PAPER WAS WHEN YOU
PUT A LOT OF IMMUNE PRESSURE ON
A CANCER, YOU'RE GOING TO SEE
PUSHBACK.
IN THIS CASE IMMUNE ESCAPE WAS
IN THE FORM OF FRAME SHIFT
MUTATION IN INTERFERON GAMMA
RECEPTOR AS WELL AS HLA LOSS.
CERTAINLY AGAIN THESE HPV
STUDIES, IF YOU GET THE RIGHT
RECEPTOR, YOU HAVE A THERAPY CAN
YOU ADMINISTER SAFELY.
THAT LED TO WHETHER WE ARE
TODAY, AND TO THE TALKS YOU'LL
HEAR TO FOLLOW ME.
THIS CONCEPT OF THE NEOANTIGEN
ANTI-T-CELL RECEPTOR.
AND THESE ARE TWO PUBLICATIONS
YOU'LL HEAR FROM MARIA AND
FRANK.
I'M GOING TO PLUG ERIC'S PAPER,
HOW TO MAKE SOME RECEPTORS
BETTER, BUT THAT'S WHERE THE
FIELD IS GOING.
AND WHERE WE ARE NOW AGAIN WE
NOW HAVE AN APPROVED T-CELL
RECEPTOR-BASED MEDICINE.
IT TOOK, AGAIN, A FEW DECADES
BUT THIS IS REPORT PUBLISHED IN
THE LANCET THIS YEAR, A COMPANY
CALLED ADAPT IMMUNE FROM THE
U.K., TREATED 52 PATIENTS IN A
MULTI-NATIONAL TRIAL IN 23
SITES, YOU CAN SEE FROM THE
WATERFALL PLOTS, SPIDER PLOTS,
THAT WHEN PATIENTS RECEIVED
THEIR ANTI-MAGE A-4 T-CELL
RECEPTOR, YOU GOT A SIGNIFICANT
OBJECTIVE RESPONSE RATE, ALMOST
40% OF SYNOVIAL SARCOMA PATIENTS
HAD OBJECTIVE RESPONSE, OVERALL
SURVIVAL NOW GOING OUT TO 17
MONTHS.
THESE ARE HEAVILY PRE-TREATED
PATIENTS.
SO REALLY AN INDICATION THAT YOU
GET THE RIGHT RECEPTOR AND
ADMINISTER IT, YOU CAN ACTUALLY
GET DURABLE CANCER REGRESSIONS.
SO IT'S BEEN A LONG JOURNEY TO
GET US WHERE WE ARE TODAY.
WE DO AGAIN HAVE THE FIRST
MEDICINE, YOU'LL HEAR THE NEXT
FEW SLIDES SOME REALLY EXCITING
WORK.
I WANT TO END TO PERSONALLY
THANK DR. ROSENBERG.
MOST OF YOU DON'T KNOW THIS, BUT
STEVE SAVED MY CAREER.
IN 2000 THE GENOME INSTITUTE
DECIDED TO CLOSE THE CLINICAL
GENE THERAPY BRANCH.
I WAS OUT OF A JOB.
I DID NOT KNOW WHAT I WAS GOING
TO DO NEXT.
AND STEVE, WHO IS ALWAYS, AGAIN,
YEARS AHEAD OF US IN THINKING,
UNDERSTOOD THAT THE TECHNOLOGY
WHICH HAD BEEN DEVELOPED IN THE
GENOME INSTITUTE TO TREAT
GENETIC DISEASES WAS WHAT HE WAS
LOOKING FOR TO BRING TO THE
SURGERY BRANCH TO DEVELOP THOSE
NEW CLASSES OF MEDICINES WHICH
YOU'VE HEARD ABOUT AND WE
CONTINUE TO HEAR ABOUT TODAY.
SO THANK YOU, STEVE.
[APPLAUSE]
LET'S HEAR SOME SCIENCE.
>> I WELCOME DR. MARIA
PARKHURST, RESPONSIBLE FOR
NEOANTIGEN WORK YOU HEARD US
TALK ABOUT TODAY.
WELCOME.
>> THANK YOU FOR THE
INTRODUCTION.
DR. ROSENBERG, I DON'T HAVE
ANYTHING VERY PARTICULAR
WONDERFULLY THING TO SAY LIKE
SOME OTHER PEOPLE HAVE, BUT
SEEING YOU FOR 30 YEARS IT'S
BEEN AN AMAZING RIDE, I'M
THANKFUL TO STILL BE ON IT.
THANK YOU FOR THAT.
TODAY I'D LIKE TO PRESENT SOME
RECENT EFFORTS AT IDENTIFYING
PERSONALIZED NEOANTIGEN REACTIVE
T-CELL RECEPTORS FROM TIL AND
USE IN PATIENTS WITH SOLID
EPITHELIAL CANCERS.
OUR STRATEGIES SHOWN HERE, WE
FIRST RESECT, DISSECT INTO SMALL
PIECES, ALLOW T CELLS TO
EMIGRATE IN THE PRESENCE OF
IL-2, AND AT THE SAME -- WRONG
WAY.
WE ISOLATE DNA AND RNA FROM
THOSE TUMOR SPECIMENS, PERFORM
SEQUENCING, IDENTIFY SOMATIC
MUTATIONS, MAKE PEPTIDES AS YOU
PREVIOUSLY HEARD, AND WE
INTRODUCE THOSE INTO AUTOLOGOUS
ANTIGEN PRESENTING CELLS,
CO-CULTURE WITH TIL, FROM TIL
FRAGMENTS, EVALUATE RECOGNITION
BASED ON INTERFERON GAMMA
SECRETION AND INSTEAD OF
SELECTING TIL FOR PATIENT
TREATMENT WE WOULD THEN SINGLE
CELL SORT INDIVIDUAL CELLS AND
SEQUENCE TO IDENTIFY INDIVIDUAL
TCR ALPHA AND BETA CHAINS.
AS ONE EXAMPLE OF AN INITIAL TIL
SCREEN THIS IS A PATIENT, 4378,
FOR THIS PATIENT WE TESTED 24
INDIVIDUAL TIL FRAGMENTS
AGAINST -- FOR RECOGNIZE OF 133
VARIANTS ENCODED IN SEVEN
CONSTRUCTS, MULTIPLE FRAGMENTS
RECOGNIZED FNG 1 IN RED STARS, 2
IN BLUE STARS.
WE DECONVOLUTED ANTIGENS,
NEOANTIGEN BY PULSING INDIVIDUAL
25 AMINO ACID PEPTIDES DERIVED
FROM TMG OF INTEREST ON THE
PATIENT'S CONTROLS AND THERE
THIS CASE IDENTIFIED REACTIVITY
ASSOCIATED WITH TMG 1 TO BE THIS
PROTEIN, MUTATED GCLM, INVOLVED
IN GLUTATHIONE SYNTHESIS.
FOR TMG 2 REACTIVITY, A PROTEIN
INVOLVED IN ALCOHOL METABOLISM.
SO ONCE WE HAD THIS INFORMATION
IN MIND, WE THEN WON'T TO SORT
THE INDIVIDUAL CELLS TO IDENTIFY
T-CELL RECEPTORS MEDIATING THOSE
REACTIVITIES.
FOR ONE FRAGMENT WE ISOLATED CD8
CELLS THAT UPREGULATED 4 11B,
AND SEQUENCING WE IDENTIFIED
SINGLE DOMINANT T-CELL RECEPTOR
SHOWN HERE, SAME FOR TMG 2
REACTIVE CELLS.
THE REACTIVE POPULATION WAS A
POPULATION OF CD8 CD4 DOUBLE
POSITIVE CELLS BUT WE ISOLATED
INDIVIDUAL 4-1BB CELLS.
ONCE WE HAD THIS INFORMATION IN
HAND, WE WANT TO GENERATE FULL
LENGTH CONSTRUCTS, WE DID THIS
BY GENERATING PROTEIN SEQUENCE,
TCR ALPHA AND BETA CHAIN REGIONS
ARE GRAFTED MURINE CONSTANT
REGIONS, SEPARATED ALPHA AND
BETA CHAINS, TO GENERATE A FULL
LENGTH T-CELL RECEPTOR ALPHA
BETA CONSTRUCTS.
THIS IS AN EXAMPLE, HUMAN MOUSE
HYBRID BETA CHAIN FOLLOWED BY IN
RED P2A SEQUENCE, FOLLOWED BY
HYBRID HUMAN-MOUSE ALPHA CHAIN,
INTRODUCED MODIFICATIONS IN
YELLOW TO ENHANCE EXPRESSION OR
TO FACILITATE CLONING.
WE CODON OPTIMIZED, CLONED INTO
RETROVIRAL VECTOR.
WE THEN PRODUCED RETROVIRUSES
ENCODING TCRs, AND DO THIS BY
INTRODUCING T-CELL RECEPTOR IN
VECTOR WITH ENVELOPE VECTOR,
INTO 293GP CELLS WITH GAGPOL
GENES, WE PRODUCE SOME
RETROVIRAL-PRODUCING CELLS AND
USE THESE AS YOU'VE HEARD TO
INTRODUCE T-CELL RECEPTOR GENES
INTO PATIENT'S PERIPHERAL BLOOD
LYMPHOCYTES.
WE CAN USE ANTI-MURINE ANTIBODY
TO EVALUATE EXPRESSION OF TCRs
WE INTRODUCE, CAN YOU SEE FOR
THE GCLM PCR WE HAVE 77%
TRANSDUCTION EFFICIENCY, AND THE
CELLS ARE EQUALLY DIVIDED
BETWEEN CD4s AND CD8s.
WE EVALUATED CELL FUNCTION BY
LOOKING AT INTERFERON GAMMA
SECRETION, IN RESPONSE TO EITHER
DENDRITIC CELLS OR MUTANT
PEPTIDE PULSED DENDRITIC CELLS,
THIS RECEPTOR CLEARLY MEDIATES
RECOGNITION OF THE MUTANT GCLM
PEPTIDE.
THIS IS THE SIMILAR DATA THAT WE
IDENTIFIED, CLEARLY HAVE WHAT WE
BELIEVE TO BE TWO GOOD CELL
RECEPTORS FOR PATIENT TREATMENT.
FOR MANY CLASS 1 RECEPTORS WE'VE
ISOLATED ELEMENTS BY PEPTIDE
PULSING MINIMAL EPITOPES
TRANSFECTED WITH EACH OF THE
PATIENTS HLA MOLECULES.
AND IN THIS CASE IDENTIFIED THAT
THE GCLM PCR RECOGNIZED THAT
PEPTIDE IN CONTEXT OF HLA 25,
ALDA 2 TCR RECOGNIZED PROTEIN IN
THE CONTEXT OF HLA 2.
SO WITH ALL THIS BACKGROUND
INFORMATION WE WERE READY TO
TRANSLATE TO GO TO THE CLINIC.
AND THERE WAS ONE HURDLE THAT
HAD TO BE OVERCOME.
FOR EVERY T-CELL RECEPTOR WE
PREVIOUSLY MADE, INTENT WAS TO
TREAT A LOT OF PATIENTS WITH A
SINGLE PRODUCT.
WE MADE T-CELL RECEPTOR
SUPERNATANTS, A SINGLE PACKAGING
CELL LINE, MAKING A MASTER BANK
OF THAT CELL LINE AND MAKING GLP
QUALITY RETROVIRAL SUPERNATANTS.
THE AMOUNT OF TIME AND COST IN
PRODUCING AND QUALIFYING THE
CELL LINE AND SUPERNATANTS MADE
IT IMPRACTICAL TO TREAT AN
INDIVIDUAL PATIENT WITH SMALL
AMOUNT OF T CELL RECEPTOR.
A HUGE EFFORT BY STEVE FELDMAN
IN THE LAB HE AND NOW RACHEL
BIER DEVELOPED A GLP PROCESS
THAT IS NOW FDA APPROVED THAT
ALLOWS US TO USE TRANSIENTLY
TRANSFECTED 293 CELLS TO PRODUCE
GLP QUALITY RETROVIRUSES, IT'S
SHOWN HERE BASICALLY ON DAY
ZERO, A VIAL FROM MASTER CELL
BANK IS THAWED, EXPANDED TO
LARGE NUMBERS, TRANSFECTION IS
DONE ON DAY 11, BY
CO-TRANSFECTING WITH RETROVIRAL
TCR CONSTRUCT, THIS RD 114 ARE
VECTOR.
IT'S HARVESTED, TREATED TO
REMOVE PLASMA, CRYOPRESERVEED.
WE TAKE PERIPHERAL BLOOD
LYMPHOCYTES FROM PATIENT, INSERT
USING GMP QUALITY RETROVIRAL
PRODUCTS, EXPAND THOSE TO LARGE
NUMBERS IN THE CLINIC USING
SECOND SIMULATION AND FINALLY
REINFUSE INTO THE PATIENT AFTER
LYMPHODEPLETING CHEMOTHERAPY.
SO, IN ORDER TO CHARACTERIZE
INFUSED CELLS WE DO A FEW SIMPLE
MANEUVERS.
CLEARLY EVALUATE THE
TRANSDUCTION EFFICIENCY LOOKING
AT EXPRESSION OF MOUSE T-CELL
RECEPTOR CONSTANT REGION.
AND THEN WE DO SOME BASIC
PHENOTYPIC EVALUATION OF THE TCR
TRANSDUCED CELLS BY LOOKING AT
CD4 AND CD8, AND CD39 AND CD69,
IN THIS PATIENT POPULATION YOU
CAN SEE ABOUT 53% OF CELLS
TRANSDUCED, A LOT MORE CD4 CELLS
THAT WERE EXPRESSING THAN CD8,
DESPITE THE FACT IT WAS A CD8
RECEPTOR.
SURPRISINGLY, VERY ENCOURAGING
FROM OUR STANDPOINT, MAJORITY OF
CELLS WERE CD39, 69 DOUBLE
NEGATIVE, A PHENOTYPE PREVIOUSLY
ASSOCIATED WITH RESPONSES IN
PATIENTS TREATED WITH TIL IN
SETTING OF MELANOMA.
THIS IS THE ALDH 2 RECEPTOR,
SAME DATA FOR THAT, MAJORITY OF
THE CELLS WERE CD 4, EFFECTOR
MEMORY PHENOTYPE BUT LARGE
PERCENTAGE, MAJORITY WERE CD39,
69 DOUBLE NEGATIVE.
WE EVALUATED FUNCTION OF THE
INFUSED CELLS LOOKING AT
RECOGNIZE OF TITRATED PEPTIDE ON
THE PATIENT'S AUTOLOGOUS
DENDRITIC CELLS, BY LOOKING AT
INTERFERON GAMMA SECRETION, YOU
CAN SEE THAT IF WE USE MINIMAL
EPITOPE IT'S RECOGNIZED DOWN TO
ABOUT ONE NANOGRAM PER MILL,
ALDL 2 RECEPTOR WAS EVEN MORE
SENSITIVE THAN THAT.
WE ALSO LOOKED AT SECRETION OF
OTHER CYTOKINES, GRANZYME B AND
ARE HIGHLY FUNCTIONAL.
AS WE HEARD TODAY, IT'S BEEN
WELL ESTABLISHED THAT METASTATIC
TUMORS CAN BE VERY HETEROGENEOUS
IN EXPRESSION OF ANTIGENS, FOR
EVERY TUMOR NOW WE ROUTINELY
EVALUATE BOTH TUMOR CELL
NEOANTIGEN CLONALITY, LOSS OF
HETEROZYGOSITY.
THIS PATIENT THAT I KEEP
REFERRING TO, WE SEQUENCED SEVEN
INDIVIDUAL FRAGMENTS FROM THIS
PATIENT, ESTIMATED THE
PERCENTAGE OF TUMOR CELLS THAT
HAD EITHER GCLM MUTATION OR ALDH
2 MUTATION, YOU CAN SEE THAT THE
ALDH 2 MUTATION APPEARS CLONAL,
100% OF THE CELLS IN EVERY
FRAGMENT THAT WE SEQUENCED HAD
THAT MUTATION.
HOWEVER, THE GCLM MUTATION WAS
SUBCLONAL, ONLY ACTUALLY FOUND
IN TWO OF THE SEVEN FRAGMENTS WE
EVALUATED.
WE FOUND NO CLEAR EVIDENCE OF
CLASS 1 HLA LOH.
THIS PATIENT DID HAVE A PARTIAL
RESPONSE TO TREATMENT SO WE HAD
TWO BIOPSIES POST TREATMENT,
MUTATION WAS CLONALLY PRESENT IN
THE ALDT 2 MUTATION IN BOTH BUT
THE GCLM MUTATION WAS
UNDETECTABLE.
THERE WAS NO EVIDENCE OF CLASS 1
LOH.
THIS INFLUENCED OUR DECISION
ABOUT HOW TO COMBINE THE TWO
TCRs FOR PATIENT TREATMENT.
WE GAVE 2/3 OF THE INFUSED
SAMPLES, ALDH 2 TCR TRANSDUCED
CELLS, A THIRD WERE GCLM
TRANSDUCED CELLS.
THIS IS A SUMMARY OF ALL THE
PATIENTS THUS FAR, IN PARTICULAR
WE'VE TREATED 11 PATIENTS, 8
WITH COLORECTAL, TWO BREAST
CANCER, ONE WITH NON-SMALL CELL
LUNG CARS NO, CARCINOMA.
AND IN FOUR PATIENTS BECAUSE WE
SAW SOMETIMES THIS EXPRESSION OF
THE TCR WE WANTED IN A T CELL
EXPRESSING OPPOSITE CO-RECEPTOR
WE SELECTED FOR APPROPRIATE IN
SOME, TREATED FOUR PATIENTS WITH
CELLS ENRICHED FOR APPROPRIATE
CO-RECEPTOR.
I DIDN'T PUT THE DATA ON THE
TABLE, WE TARGETED ONE CLONAL
NEOANTIGEN, DIDN'T TREAT ANY
PATIENTS WITH CLASS 1 LOH
RELEVANT TO THE RESTRICTION
ELEMENT ASSOCIATED WITH TUMOR
TREATMENT.
MOST OF THE PATIENTS GOT ON THE
ORDER OF ONE TIMES 10 TO THE
11th CELLS, AND AT LEAST 50%
TRANSDUCED.
AND MAJORITY OF CELLS, EVERY
PATIENT HAD EFFECTOR MEMORY
PHENOTYPE, FORTUNATE WERE ABLE
TO GIVE EVERY PATIENT AT LEAST
10% USUALLY GREATER OF THESE
CD39 CD69 DOUBLE NEGATIVE CELLS,
TEN TO 100 TIMES THE PERCENTAGE
OF CELLS WE NORMALLY GIVE WITH
SELECTED TIL THAT HAD THIS
POPULATION.
THREE PATIENTS HAD PARTIAL
REJECTIVE RESPONSES BY RECIST
CRITERIA, 4 TO 7 MONTHS, ONE
RECEIVED A CD4 T-CELL RECEPTOR.
WE'VE ALSO TREATED SEVEN
PATIENTS PREVIOUSLY TREATED WITH
TIL OR ANOTHER T CELL-BASED
THERAPY TARGETING THE SAME
NEOANTIGENS WE DID IN OUR SECOND
THERAPY WITH THE TCRs.
AND WE DIDN'T SEE ANY RESPONSES
IN THIS GROUP, AND WE SPECULATE
THIS MAY LIKELY BE THE FACT WE
TARGETED THE SAME NEOANTIGENS
INEFFECTIVELY TARGETED IN THE
FIRST TREATMENT.
WE'VE TREATED THREE PATIENTS
WITH BREAST CANCER,
COMPASSIONATE EXEMPTION,
UNFORTUNATELY NONE OF THOSE
PATIENTS RESPONDED TO THERAPY.
IN SOME PATIENTS WE EVALUATED
PERSISTENCE OF TRANSDUCED CELLS,
IN PERIPHERAL CIRCULATION, WE
DISTINGUISHED BETWEEN THE TWO
BECAUSE THERE'S AN ANTIBODY
AVAILABLE.
THIS IS WHAT PRE-TREATMENT LOOKS
LIKE, DAY 7 POST TREATMENT 15%
EXPRESSED GCLM TCR, REPRESENTING
THE WAY INFUSION WE PROVIDED TO
THE PATIENT.
WE EVALUATED PERSISTENCE IN THE
FIRST SEVEN PATIENTS, THUS FAR.
AND IF WE LOOK AT THE
NON-RESPONDERS YOU CAN SEE TWO
OF THE NON-RESPONDERS, IN TWO
NON-RESPONDERS THE TCR
TRANSDUCED CELLS DWINDLED AND
WERE NOT APPARENT BEYOND ABOUT
DAY 40 AFTER TREATMENT.
BUT FOR TWO OF THE OTHER
NON-RESPONDERS WE COULD READILY
DETECT CELLS, YOU CAN SEE THE
LAST PATIENT WHO 50 DAYS STILL
HAD 60% OF CIRCULATING CDs 3
CELLS EXPRESSING T-CELL RECEPTOR
WE GAVE HER.
WHEN WE LOOKED AT RESPONDERS WE
HAVE LARGER TIME POINTS
AVAILABLE FOR RESPONDING
PATIENTS, WE COULD DETECT
TRANSDUCED CELLS IN EVERY
PATIENT, AT EVERY TIME POINT WE
EVALUATED.
AND I'LL SHOW A FEW EXAMPLES OF
RESPONSES.
THIS PATIENT HAD REGRESSION OF
LEFT LUNG LESION AS WELL AS
MEEDALSTINAL LYMPH NODES, THIS
HAD RETRO PERITONEAL, DOCUMENTED
LUNG DISEASE, RETRO PERITONEAL
LYMPH NODES AND PERIHEPATIC
TUMORS.
SO, I WAS GOING TO PRESENT A
LITTLE BIT OF ADDITIONAL DATA
ABOUT PATIENTS WHOM WE ATTEMPTED
TO TARGET T CELLS AGAINST THESE
COMMON DRIVER MUTATIONS, BUT DR.
ROSENBERG PRESENTED THAT, WE
DON'T NEED TO GO INTO THAT
FURTHER.
WE NOW HAVE A LIBRARY OF 39
REACTIVE T-CELL RECEPTORS, 23
KRAS, AND WE HAVE TREATED 26
PATIENTS WITH A VARIETY OF
THESE.
THESE ARE PATIENTS THAT COME TO
US WITHOUT HAVING TO DO A
RESECTION, IF WE HAVE SEQUENCING
INFORMATION, IF THEY HAVE THAT
PARTICULAR HOT SPOT MUTATION AND
APPROPRIATE HLA RESTRICTION
ELEMENT WE CAN GIVE AN OFF THE
SHELF REAGENT QUICKLY.
T CELLS THAT RECOGNIZE UNIQUE
SOMATIC MUTATIONS CAN BE FOUND
IN TIL.
FROM THE COMMON EPITHELIAL
CANSERS IS AND TCRs THAT
MEDIATE CAN READILY BE
IDENTIFIED, ISOLATED, USED TO
GENETICALLY MODIFY PBL.
CELL TRANSFER THERAPY WITH PBL
TO EXPRESS PERSONALIZED
NEOANTIGEN RECEPTORS, CAN
MEDIATE REGRESSIONS IN PATIENTS
WITH COLORECTAL CANCERS
REFRACTORY TO ALL OTHER
TREATMENTS.
WE HAVE NOT IDENTIFIED ANY
CORRELATES WITH CLINICAL
RESPONSE BUT AS YOU'LL PROBABLY
HEAR THROUGHOUT THE FEW DAYS A
LOOT OF PEOPLE ARE PURSUING
STRATEGIES TO IMPROVE THE
THERAPY, AND I WOULD SAY ONE
MAJOR EFFORT OR TWO MAJOR
EFFORTS ONGOING IN THE LAB THAT
ARE REALLY DESIGNED TO OVERCOME
THESE BARRIERS OF EXPRESSION AND
LOSS OF HETEROZYGOSITY TO
IDENTIFY MORE T-CELL RECEPTORS
FOR EACH PATIENT AND DEVELOP
PRACTICAL METHODS FOR TREATING
PATIENTS WITH THESE MORE DIVERSE
T CELL POPULATIONS, AND I THINK
DR. LOWERY AFTER ME WILL
DESCRIBE THESE EFFORTS.
THANK YOU VERY MUCH FOR YOUR
KIND ATTENTION.
[APPLAUSE]
>> DR. FRANK LOWERY WILL TALK
MORE ABOUT HOW WE'RE BROADENING
OUR TCR HUNT.
HE SERVES RESEARCH CAPACITY BUT
MOVING THINGS FROM THE LAB INTO
OUR GMP SPACES.
>> THANK YOU, DR. GOFF.
IT'S AN HONOR TO BE HERE.
THANK YOU TO DR. PARKHURST FOR
GIVING CONTEXTUALIZATION, AND
FOR TAKING ME UNDER YOUR WING
WHEN I GOT HERE AND SHOWING -- I
CAME IN WITH NO IMMUNOLOGY
BACKGROUND, A CANCER BIOLOGIST
IN GRADUATE SCHOOL, AND MARIA
SHOWED ME THE COMPLICATED NATURE
OF OUR T CELL ASSAYS, HOW TO
DISCOVER NEOANTIGENS, CLONE
TCRs, APPRECIATIVE TO HER FOR
THAT.
SO ANYONE WHO HAS EXPERIENCE IN
PATIENT CARE OR FAMILY MEMBER OR
DEALT WITH CANCER, MAJOR
LIMITING FACTOR IN DEVELOPING
THESE SPECIALIZED LAST DITCH
THERAPIES FOR PATIENTS DEALING
WITH ADVANCED METASTATIC DISEASE
IS TIME.
THAT'S THE ONLY NON-NEGOTIABLE.
SO WHEN WE'RE STARTING THE CLOCK
ON OUR TCR HUNT FOLLOWING TIL
SCREEN, ALREADY AN SEQUENCE
RESOURCE INTENSIVE AND TIME
INTENSIVE SEARCH, WHEN WE ONLY
START LOOKING FOR TCR FOLLOWING
TIL SCREENS, IT MEANS WE'RE
ADDING TWO MONTHS OR MORE OF
LOST TIME TO THAT CLOCK THAT THE
PATIENT'S DON'T HAVE HAVE TO
SPARE.
ONE POINT TO MAKE, IF WE GO
HUNTING FOR TCRs BASED OFF TIL
SCREEN DATA, OUR TCR TARGETS ARE
ONLY GOING TO BE LIMITED TO
THOSE THAT WE OBSERVED IN
FUNCTIONAL REACTIVITY SCREEN OF
THE TIL.
SO IF THE TIL WERE TOO EXHAUSTED
TO SHOW FUNCTIONAL REACTIVITY WE
WON'T PICK IT UP.
IF THE REACTIVITY WAS TOO WEAK
WE MIGHT MISS THAT WAY.
WE'LL BE LIMITED IN OUR
POTENTIAL NUMBER OF TARGETS WE
CAN HUNT THAT WAY.
AND THAT WHEN WE FIND THE
POTENTIAL CANDIDATE TCR THAT
SEEMS TO BE TUMOR MUTATION
REACTIVE, THE IDENTIFICATION,
SEQUENCING, THAT'S ONLY THE
FIRST STEP.
ULTIMATELY WE HAVE TO VALIDATE
MUTATION SPECIFIC, WE'RE SO
LUCKY TO BE HERE AT SURGERY
BRANCH, BUT THE GMP
QUALIFICATION, PREP OF A
PERSONALIZED CANCER TREATMENT IS
ANOTHER RESOURCE INTENSIVE PART
OF THE PROCESS.
SO, ONE MAJOR QUESTION THAT WE
SET OFF ON, AND ORIGINAL HUNT
WORK WAS WITH DR. KRISHNAN, MY
PARTNER IN CRIME IN A LOT OF
THIS STUFF.
ONE QUESTION WAS CAN TUMOR
RELEVANT TCRs BE PREDICTED
BASED ON SHARED PHENOTYPES?
ONE DUTY IS TCRs CAN BE USED
IF IT'S A TUMOR-SPECIFIC TCR YOU
CAN INTRODUCE IT INTO A
SECONDARY LYMPHOCYTE AND USE IT
TO REDIRECT SPECIFICITY TOWARDS
YOUR TARGET OF CHOICE.
THE OTHER BEAUTY AS A RESEARCHER
THAT THE TCR SERVES AS BARCODE
AND CAN YOU HAVE THAT BE A
BENCHMARK FOR THE CELL OF
INTEREST THAT EXPRESSES THAT
TCR.
ALL IT TAKES IS THE ROSETTA
STONE HAVING THE TCR REACTIVITY
DATA ALLOWS YOU TO TRACK THE
CLONES THAT HAVE THE SAME
REACTIVITY.
SO BASICALLY THAT'S THE
BACKGROUND IN HOW WE WERE ABLE
TO USE RNA SEQUENCING ANALYSIS
TETHERED TO FIND SHARED REACTIVE
PHENOTYPES.
AS A BRIEF BACKGROUND I DON'T
WANT TO GO INTO THE PUBLISHED
WORK, BUT OUR STUDY IN 202
SHOWED MAJORITY OF NEOANTIGEN
REACTIVE TCRs, THAT MARIA HAD
FOUND, I WAS LUCKY ENOUGH TO
SHARE MY WORK IN SORTING TO GET
THIS, BUT WE HAD BUILT A BANK OF
TCRs FOR PATIENTS, WITH
PRESERVED AVAILABLE WE STUDIED
THEM, ASKED THE QUESTION WHAT
STATE WERE THE CELLS BEARING
NEOANTIGEN REACTIVE CELLS IN.
WE ANALYZED TEN PATIENTS, NINE
HAD KNOWN REACTIVITY, WE SAW
THERE WAS REMARKABLE BIAS IN
LOCATION OF THE KNOWN NEOANTIGEN
REACTIVE TCRs, CD 4s IN THIS
STATE, CD8S IN EFFECTOR-LIKE
STATE.
WE BEEFED UP KNOWLEDGE, WHAT
ABOUT OTHER CELLS IN THE
CLUSTERS, REPREDICTED MORE
REACTIVITY BASED ON THE SHARED
PHENOTYPE AND ARCHIVAL SAMPLES,
AND SHOWED INDEED WHEN YOU
LOOKED FURTHER INTO THESE
CLUSTERS YOU FOUND MORE CD8
REACTIVE HERE AND MORE CD4
REACTIVE NEOTCRs HERE.
ONCE WE ASSEMBLED THE DATABASE,
WE LOOKED AT DIFFERENTIALLY
EXPRESSED GENES OF THE TUMOR
SPECIFIC CELLS, PERFORMED ROC
ANALYSIS.
IDENTIFIED WHAT WE CALLED NEOTCR
SIGNATURE, SENSITIVE FOR CD4
CELLS IN OUR STUDY, AND 8
SIGNATURE FOR CD8s.
NOTABLY THERE BE WERE SHARED
GENES AMONG THESE CD4 AND 8
STATES, NOT EXPRESSING MEMORY
STEM-LIKE MARKERS.
NOW, WHEN WE COMPARED OUR
SIGNATURES TO STUDIES DONE IN
OTHER TYPES OF TUMOR SOLID
GENES, ET CETERA, BASICALLY
PEOPLE AROUND THE WORLD OBSERVED
THE SAME PHENOMENON IN BLADDER
CANCER, LUNG CANCER, MELANOMA,
ET CETERA.
OUR SHARED STATE IS A COMMON
PHENOMENON.
IF WE LEARN THIS KNOWLEDGE IN
ARCHIVAL HOW CAN WE USE THIS
GOING FORWARD?
IF WE HAVE A PATIENT TUMOR IS
RESECTED, IMMEDIATELY START
HUNTING FOR TCR, WE TAKE FRESH
TUMOR DIGEST, ISOLATE, WE'RE NOT
JUST SEQUENCING ENTIRE TUMOR,
ARTIFICIALLY ENRICHING THE
LYMPHOCYTE COMPONENT.
WE PERFORMED SAME TRANSCRIPTOMIC
CLUSTERS ON THE ARCHIVAL SAMPLES
BUT NOW WE OBVIOUSLY HAVE SOME
BIAS IN OUR SIGNATURES.
THE FIRST STEP IS LOOK FOR
CLUSTERS THAT SELF-ASSEMBLE,
AROUND CAN YOU PROJECT ON THE
CLUSTERS AND MARK HOT REGIONS
THAT ARE A GOOD FIT FOR
SIGNATURES.
AND THEN BASED ON THAT YOU CAN
RECONSTRUCT AS DR. MORGAN TALKED
ABOUT, THE ALPHA BETA PAIR,
EXPRESS IN DONOR PLBR, SECRETARY
TO THE SAME PERSONALIZED
SCREENING METHODS WE'VE USED FOR
TIL.
AND IF YOU FIND VALIDATED
REACTIVE ONE THAT CAN BE A
TREATMENT CANDIDATE TCR.
I WANT TO SHOW HOW THIS LOOKS IN
PRACTICE, BUT IF YOU GETS MORE
IN THE WEEDS, NO PRIOR KNOWLEDGE
OF SAMPLE COMING IN, YOU NEED TO
PERFORM TRANSCRIPTOMIC
CLUSTERING, USE THE BIOCONDUCTOR
ALGORITHM, SCORES INDIVIDUAL
CELLS FOR FITNESS TO A GIVEN
SIGNATURE.
YOU'RE SEEING HOW WELL CELLS IN
PATIENT A'S TUMOR MAPS TO KNOWN
PREVIOUSLY DESCRIBED SIGNATURES,
WITHIN THAT POPULATION, AGAIN NO
IDEA OF REACTIVITY, WE DON'T
HAVE THE PATIENT'S TUMOR
SEQUENCING DATA YET, THIS SET OF
CELLS HAS A GREAT FIT FOR NEO8,
CD8 REACTIVE TO THE PATIENT'S
TUMOR ANTIGENS, WE ALSO HAVE THE
OPTION CITIED OF LIMITING TO
CELLS DEMARCATED CAN LOOK AT
CLUSTERS THAT SELF-ASSEMBLE,
I'LL SHOW WHAT I MEAN IN A
SECOND.
OCCASIONALLY UNAMBIGUOUS,
CLUSTER DEFINES ITSELF, WE
SCREEN AGAINST PATIENT
TUMOR-SPECIFIC REAGENTS,
PEPTIDES.
TMGS, TUMOR ORGANOIDS WHEN
AVAILABLE.
HERE IS ONE EXAMPLE, A COLON
CANCER, LIVER METASTASIS.
GENERALIZED CLUSTERING.
I SHOWED SELECTED GENES HERE,
I'M SORRY IT'S SMALL, UPPER
RIGHT PART OF THE MAP IS LARGELY
CD4.
LOWER INTO THE LEFT CD8.
AS THE BRIEFEST SHORT HAND FOR
SIGNATURES, CXCL 13 IS ONE GENE
THAT'S SHARED, HIGHLIGHTING THIS
REGION HERE, THIS REGION HERE.
YOU HAVE TO -- THE HUMAN MIND
ISN'T DESIGNED TO JUMP BACK AND
FORTH.
IF YOU NOTE REGIONS HIGHLIGHTED
IT WOULD BE CLUSTER DOWN HERE,
THE POSITIVE REGION, AND C17 IS
THIS TINY CLUSTER DEMARCATED
HERE.
WHEN PROJECTED ON THE MAP,
PRETEND YOU DON'T KNOW THE
CLUSTERING IDENTITY, YOU LOOK
FOR HOT SPOTS, YOU SAY ON THE
NEO4 SIGNATURE A HOT SPOT HERE,
HERE, HERE.
NOW, NEO8 SIGNATURE HIGHLIGHTS
ONE REGION, SO NEO4 IS NOW
PICKING UP CLUSTER THAT I WOULD
SUGGEST IS CD8 CLUSTER.
CXCL 13 POSITIVE UP HERE, ONE
ADDITIONAL CLUSTER, TREG,
DEMARCATED BY FOXP3, THREE HOT
SPOTS FOR CD4 SIGNATURE, AND I'M
PROJECTING ONES VALIDATED TO BE
TUMOR MUTATION SPECIFIC.
IN THIS CASE ONLY ONE OUT OF 13
PREDICTED CD4s WAS REACTIVE
BUT DID LIVE WITHIN CLUSTER 17,
IN FACT A SINGLE CELL WAS HIGHLY
SCORING FOR SIGNATURE, NOT A
CLONALLY EXPANDED CELL BUT
SEQUENCING AGAINST RANDOM MU.
TEN WERE REACTIVE.
THE VAST BIAS WHERE CELLS
HOUSING REACTIVE CELLS WERE IN
CLUSTER 9, WE DON'T SEE A HOT
SPOT IN ADDITIONAL CLUSTER,
EXCEPTIONAL CASINGS OF CELLS
BEARING TCRs FOUND IN OTHER
STATES.
SIGNATURE IS NOT A RULE.
YOU'RE CAPTURING CELLS IN
MULTIPLE STATES OF EXHAUSTION OR
ACTIVATION, BUT BY AND LARGE
THEIR FOCAL POINT IS DOWN HERE.
WE PERFORMED THE SAME ANALYSIS
IN SECOND PATIENT, RECTAL CANCER
LUNG METASTASIS.
THIS HAD LESS OF A CLEAR
DEMARCATION PATTERN WHERE THE
REACTIVITY WAS HOUSED, TINY
CLUSTER, CXCL 13 HOT, CD8 HOT
RIGHT HERE.
AND THEN A REGION OF THIS
CLUSTER THAT WAS POSITIVE UP
HERE, NOT EVEN A LARGE REGION
WHATSOEVER.
LOOKING AT SIGNATURE PROJECTIONS
THIS IS MORE CONFUSING, NOT ONE
CLEAR ANSWER WHERE REACTIVITIES
MIGHT HAVE BEEN HOUSED.
SO WE THINK THIS IS A CHANCE,
WE'RE LUCKY TO BE HERE, THIS IS
A CHANCE TO LEARN MORE.
YOU STRETCH THE LIMIT SOMETIMES
AND IF IT'S AM AMBIGUOUS YOU ME
MORE TCRs.
I MADE THEM FROM THIS REGION
RIGHT HERE SCORING HIGHLY FOR
NEO8 SIGNATURE HERE.
FROM THIS FORK UP HERE THAT
SCORED HIGHLY FOR NEO4, AND THEN
ADDITIONALLY FOR CELLS SCATTERED
IN DIFFERENT STATES, IMPERFECT
FIT FOR OUR KNOWN SIGNATURES BUT
MIGHT BE FRUITFUL IF WE GO
LOOKING.
HERE IS VALIDATED TCRs.
32 PREDICTED, 9 REACTIVE TO 5
NEOANTIGENS.
THREE VERSES ONE, TWO VERSES
ANOTHER, TWO VERSUS ANOTHER, AND
THEN ONE VERSUS ANOTHER RANDOM
MUTATION.
YOU CAN SEE CD4s ARE
INTERESTING IN THAT A FEW OF
THEM WERE IN THAT FORK AREA
WHERE I PREDICTED CD4 REACTIVE
CELLS WOULD BE BUT A NUMBER WERE
CO-CLUSTERING WITH CD8 POSITIVE
CLUSTER, BY SIGNATURE THEY
SCORED HIGHLY.
THERE WAS A CLONE FOUND IN THE
TREG CLUSTER AT LEAST ONE, SAME
HERE WITH REACTIVE CD8 SAW --
SORRY, GETTING CUT OFF AT THE
BOTTOM.
WE SAW OF 24 PREDICTED CD8S, 14
REACTIVE, TO VAN GOGH LIKE 1,
ORPHAN TCR REACTIVE TO ORGANOID.
CD8s WERE MORE CENTRALLY
LOCATED RIGHT HERE IN THIS ONE
REGION.
LAST IS ANOTHER RECENT SAMPLE.
THIS WAS CONFUSING THERE WAS NO
GREAT HOT SPOT CLUSTER, DIDN'T
HAVE 103, CXCL 13, CD39 IN THE
SAME REGION.
JUST AS A QUICK CHECK YOU CAN
SEE CXCL 13 IS IN THE SOUTHERN
PORTION OF THIS LARGE CLUSTER.
CD8s ARE PROBABLY MORE POORLY
DEFINED HERE BUT IN GENERAL TO
THE RIGHT-HAND SIDE OF THE MAP.
CD4s ARE LARGELY OVER HERE, AT
THE GENE EXPRESSION LEVEL ON AN
INDIVIDUAL CELL MIGHT BE HARD TO
PICK UP CD4s BY SINGLE CELL
SEQUENCING.
WE PROJECTED THE SIGNATURES NOW,
CD8s ARE IN THIS REGION OVER
HERE, THAT DIDN'T HAVE CXCL 13,
A PARTIAL FIT FOR THE SIGNATURE,
THE POINT OF HAVING A DEEP
MULTIPLEX 240 GENE SIGNATURE,
NOT EVERY SAMPLE WILL HAVE THE
BEST FIT.
CD8s PREDICTED TO BE HERE,
CD4s HERE, TREG CLUSTERS UP
HERE.
WHEN WE PREDICTED THESE AGAIN WE
WERE VERY LOOSE ABOUT OUR
DEFINITION OF WHAT MIGHT BE A
HIT HERE BECAUSE WE WANTED TO
PUSH LIMITS.
FOR CD4s PREDICTED 30 TCRs,
4 REACTIVE.
s CD4s WERE NOT TIGHTLY
LOCATED HERE.
IF WE MADE IT BASED ON THAT
CLUSTER THAT WAS A CXCL 13 HERE
WOULD HAVE MISSED THESE GUYS,
BASED ON THE SIGNATURE, PEPPERED
THROUGHOUT THIS CD4 REGION.
CD8s WE PREDICTED 41 TCRs,
ONE SHOWED REACTIVITY.
AND INTERESTINGLY ENOUGH THIS
CD8 TCR IN THIS REGION, AGAIN IT
WAS AVOIDING CXCL 13 HOT REGION
HERE, IN AN INTERESTING
TRANSITIONAL STATE WHERE IT HAD
CD103, NOT CD39, INTERESTINGLY
ENOUGH THIS IS THE SAME TCR
MARIA FOUND THROUGH TIL
SCREENING FOR THIS PATIENT.
THAT MIGHT BE NOTEWORTHY.
SO I SHOWED THREE EXAMPLES.
WE'VE, KRISHNAN AND MY GROUP,
COMPLETED SCREENING OF EIGHT
ADDITIONAL SAMPLES THAT
COLORECTAL, BREAST, PANCREATIC
CANCER, SIX ADDITIONAL SAMPLES
IN THE SEQUENCING PIPELINE.
I HOPED TO SHOW TCR CLUSTERS CAN
BE OBVIOUS AND WELL DEFINED OR
VERY AMBIGUOUS.
IF I HAVE A FEW MINUTES LEFT, I
WANT TO RETROSPECTIVELY
REANALYZE NEW SAMPLES SINCE
PUBLICATION BASED ON PATIENTS
SINCE 2023.
AND REVISIT THE QUESTION DO
THESE PREDICTED TCRs HAVE
SIMILAR PHENOTYPE TO INDEPENDENT
SAMPLES PREVIOUSLY, SO WE
CO-CLUSTERED T CELLS FROM 11
SAMPLES WITH REACTIVITY AS WELL
AS TWO SAMPLES FROM PATIENTS TIL
HARVESTS IN THE PAST MONTH.
CO-CLUSTERED INTO ONE UMAP, 22
CLUSTERS OVERALL.
PROJECT OUR VALIDATED NEOANTIGEN
REACTIVE CELLS THEY FELL WITHIN
SEVERAL DISTINCT REGIONS, NOT
JUST THE TWO THAT WE HAD COME TO
EXPECT PREVIOUSLY.
AND YOU CAN SEE ALL TIL
BREAKDOWN IS HERE, YOU GENERALLY
HAVE YOUR LOWEST NUMBERED
CLUSTERS, TOP RANKED, HERE A
CLEAR BIAS C3 IS AN AREA, A HOT
SPOT FOR CD8s, C4 ANOTHER HOT
SPOT FOR CD8s.
C6 IS THE TREG CLUSTER, IN THIS
CASE INCLUDES SOME CD8 REACTIVE
CELLS THAT EXPRESS FOXP3, C 16
TRADITIONAL CLUSTER, INTERESTING
HERE NOTABLE CLUSTER WAS C14,
CXCL 13 CLUSTER, THIS IS SHOWING
THE TWO YET TO BE TESTED
SAMPLES, 4632 IS COLORECTAL,
LUNG METASTASIS, NICELY
REPRESENTED HOT SPOT IN C3 WITH
REACTIVITY IN OTHER SAMPLES.
AND 4632 COLORECTAL TO LIVER
METASTASIS, A NICE REGION HERE
AS WELL AS THIS GRANULATION
POSITIVE CXCL 13 REGION HERE.
HERE I'VE REPRODUCED THE SAME
FIGURE IN THE ORIGINAL PAPER, IF
WE LOOK AT GENE EXPRESSION, YOU
CAN SEE CXCL 13 IS POSITIVE
AMONG MOST REGIONS OF REACTIVITY
BUT NOTABLY ABSENT HERE.
CD39 IS LOWER HERE.
THE REGION IS TOX POSITIVE, NOT
LIKE THESE ARE STEM LIKE CELLS
BUT THEY DO NOT HAVE THE SAME
DYSFUNCTIONAL PHENOTYPE WE'RE
USED TO SEEING IN ALL THE
SAMPLES.
NOW IF YOU COMPARE OUR
SIGNATURES TO EVERYONE ELSE'S
SIGNATURES, THERE IS A NICE
OVERLAP OF A CONSERVED PROGRAM
THAT'S NOT 243 GENES LONG, ONLY
21 GENES LONG FOR CD8S, VERY FEW
COMMON TO ALL CD4s, SHOWING
OUR SIGNATURE NEEDS REFINEMENT
STILL.
WE WANT IT TO BE A CATCH-ALL BUT
CLEARLY IT'S HARD TO BE A
CATCH-ALL IN EVERY INDIVIDUAL
SAMPLE.
SO THIS IS WHERE WE'RE GOING TO
PUT FURTHER EFFORT INTO THE
FUTURE.
DETERMINING WHAT THE BEST
VERSION OF THIS SIGNATURE IS.
SO IN CONCLUSION THE NEOANTIGEN
SIGNATURES CAN BE USED TO
PREDICT RELEVANT TCRs, WE DO
SEE BOTTLENECKS IN THE CANCER
IMMUNOGENICITY WHERE WE FIND
REDUNDANCY, YOU MIGHT FIND A
TOTAL OF ONLY TWO TARGETS,
ANOTHER SPACE TO LOOK INTO DR.
GROSS'S WORK ON DARK MATTER, IF
THEY HAVE THIS EXHAUSTION
SIGNATURE WHY AREN'T THEY
RECOGNIZING OUR ANTIGENS.
THIS IS ALL LIVE ANALYSIS,
ONGOING ON CURRENT CLINICAL
SAMPLES, KNOWLEDGE CONTINUES TO
EVOLVE.
TO ACKNOWLEDGE DR. ROSENBERG, AN
HONOR TO HAVE GOTTEN TO WORK
WITH YOU.
YOU'RE NOT SUPPOSED TO MEET YOUR
HEROES, IF YOU GET THE PRIVILEGE
TO WORK WITH ONE, IT'S BEEN
AMAZING.
THANK YOU.
MY TEAM, THANK YOU FOR ALL THE
WORK.
AND KRISHNA AND LAB MEMBERS,
THANK YOU FOR THE SCREENING OF
ADDITIONAL SAMPLES.
AND PARTICULARLY SIVASISH FOR
HIS WORK.
WE'RE SHORT ON TIME.
IF YOU HAVE QUESTIONS, PLEASE
E-MAIL.
THANK YOU.
[APPLAUSE]
>> THANK YOU.
I'D LIKE TO INTRODUCE DR. ERIC
TRAN.
WE HAVE CAUGHT UP A LITTLE BIT
BUT I WOULD ASK THAT AT THE
CONCLUSION WE'LL HAVE A
10-MINUTE BREAK RATHER THAN 15.
HERE REGO.
HERE WE GO.
>> IT'S WONDERFUL TO BE BACK ON
CAMPUS.
IT'S AN ABSOLUTE HONOR TO BE
HERE CELEBRATING DR. ROSENBERG'S
50 YEARS AT NCI.
I KNOW MOST OF YOU THINK 50
YEARS OF RESEARCH IS A LONG TIME
BUT IF YOU ASK HIM HE WILL SAY
THAT HE FEELS LIKE HE'S JUST
GETTING STARTED.
WITH THAT, HERE WE GO.
I'M GOING TO SWITCH GEARS A
LITTLE BIT AND I KNOW I'M GOING
TO DISCUSS A PROJECT WHERE IF
YOU GOT A GOOD T-CELL RECEPTOR,
A GOOD TIL THAT TARGETS A
NEOANTIGEN WHAT CAN WE DO TO
ENHANCE THE T CELLS.
HERE ARE MY DISCLOSURES.
HERE IS GENERAL STRATEGIES OF
THE FIELD, PURSUING TO ENHANCE
CELLULAR THERAPIES.
A MAJOR ADVANTAGE IS THAT HAVE
YOU THE ABILITY TO MANIPULATE
THE CELLS EX VIVO, YOU CAN
MANIPULATE THE HOST IN VIVO.
AND SO I'M NOT GOING THROUGH IN
TOO MUCH DETAIL HERE BUT WE CAN
MANIPULATE CELL MANUFACTURED
CONDITIONS TO GENERATE POTENT T
CELLS, YOUNGER STEM-LIKE CELLS,
LEVERAGE CELLS TYPES, FOCUS ON
ALPHA-BETA T CELLS BUT OTHERS
MIST BE ADVANTAGEOUS.
HARNESSING RECEPTORS THAT TARGET
MULTIPLE ANTIGENS MAY OVERCOME
TUMOR HETEROGENEITY.
WE CAN GENE ENGINEER CELLS AND
KNOCK OUT GENES THAT INHIBIT T
CELL ACTIVITY.
WE CAN KNOCK IN GENES THAT
ENHANCE T CELL ACTIVITY.
IN VIVO SIDE, HOST
LYMPHODEPLETION APPEARS TO BE
IMPORTANT TO ENHANCE CELL
THERAPY, WE CAN DELIVER
CYTOKINES, CHECKPOINT BLOCKADE,
IMMUNE AGONIST, VACCINES, TUMOR
TARGETING SMALL MOLECULES,
AGENTS THAT MODULATE MYELOID AND
TREGS.
TODAY I'M GOING TO FOCUS ON ONE
WORK IN PROGRESS PROJECT THAT
DEALS WITH CHIMERIC RECEPTORS.
MOST PEOPLE THINK ABOUT CHIMERIC
ANTIGEN RECEPTORS, HIGHLIGHTED
ON THE LEFT.
THESE TARGET ANTIGEN, DELIVER
T-CELL ACTIVATION SIGNAL.
I'M NOT FOCUSING ON CHIMERIC
ANTIGEN RECEPTORS BUT
NON-ANTIGEN RECEPTORS.
HERE IS AS EXAMPLE, SWITCH
RECEPTORS, YOU REQUEST HAVE A
CHIMERIC RECEPTOR WITH AN
EXTRACELLULAR DOMAIN FUSED.
BY BINDING THE T CELL GETS A
POSITIVE SIGNAL INSTEAD OF
NEGATIVE SILL NAIL. -- SIGNAL.
THE TYPE OF IS A CCR.
THERE ARE SEVERAL EXAMPLES OF
CHIMERIC CYTOKINE RECEPTORS,
HIGHLIGHTED HERE, SO
EXTRACELLULAR DOMAINS MADE UP OF
IL-4 RECEPTOR, TGF-BETA
RECEPTOR, STITCHED ON TO
INTRACELLULAR SIGNALING DOMAINS.
THINKING ABOUT THIS CONCEPT,
WHAT ARE THE KEY CHARACTERISTICS
THAT MIGHT MAKE AN IDEAL
CHIMERIC CYTOKINE RECEPTOR.
ONE OF THE CHARACTERISTICS IS
THAT EXTRACELLULAR DOMAIN BINDS
TO CYTOKINE NOT BENEFICIAL FOR T
CELL FUNCTION U DON'T WANT TO
TAKE A POSITIVE SIGNAL FROM T
CELL BUT POTENTIALLY NEUTRAL OR
NEGATIVE SIGNAL TURNING INTO A
POSITIVE.
REGULATED, NOT EXPRESSED, AND
RESISTANT TO TUMOR
MICROENVIRONMENT CONDITIONS SUCH
AS ACIDIC ENVIRONMENT.
INTRACELLULAR DOMAIN SHOULD
DELIVER A SIGNAL THAT PROMOTES T
CELL FUNCTION.
SO HOW DO WE IDENTIFY SUITABLE
CYTOKINES TO TRIGGER RECEPTOR?
ONE OF THE THINGS WHEN WE HAVE
CLINICAL TRIALS WE'RE ABLE TO
TREAT PATIENTS AND ANALYZE THEIR
SERUM.
SO I'M GOING TO FOCUS ONE OF THE
FIRST THINGS WE LOOKED AT WAS
LOOKING AT CYTOKINES IN THESE
PATIENTS AFTER CELL TRANSFER.
SO, THE PATIENTS ARE FROM THIS
PUBLICATION TWE YEARS AGO, MET
METASTATIC PANCREAS CANCER,
SUMMARIZING THOSE TWO PATIENTS
HERE IN THIS TABLE.
BOTH RECEIVED LYMPHODEPLETION
REGIMEN, 0361 GOT SURGERY BRANCH
TYPE LYMPHODEPLETION, 4483
SLIGHTLY DOSE REDUCED REGIMEN,
BOTH RECEIVED GOOD NUMBER OF
CELLS, CLOSE TO 30 BILLION IN
ONE CASE, 16 BILLION IN THE
OTHER.
WE SUPPORTED THOSE TRANSFERRED T
CELLS WITH HIGH DOSE
INTERLEUKIN-2.
FOR ONE PATIENT BEST RESPONSE
LASTED ONE MONTH, THE OTHER BEST
RESPONSE WAS P.R. 72% REDUCTION
IN METASTATIC TUMORS FOR SIX
MONTHS, LASTED 12 MONTHS.
PERSISTENCE OF CELLS WAS GOOD.
SO ABOUT 90% FOR FIRST PATIENT,
AT ONE MONTH.
13% FOR THE OTHER PATIENT.
BUT THE FACT THAT THEY DID NOT
ACHIEVE DURABLE COMPLETE
RESPONSE MEANS WE HAVE ROOM TO
IMPROVE TO MAKE THIS THERAPY
BETTER.
THE CELLS ARE PERSISTENT,
SUGGESTING WE MIGHT BE ABLE TO
BOOST THEIR FUNCTION.
THESE ARE THE TWO PATIENTS
SAMPLES SERUM CYTOKINE DATA.
WE FOCUSED ON T CELL EFFECTOR
CYTOKINE.
THESE ARE THE TWO PATIENTS SHOWN
HERE, THESE ARE THE CLASSIC T
CELL EFFECTOR CYTOKINES, AND YOU
CAN SEE THAT IN BOTH PATIENTS
THESE EFFECTOR CYTOKINES ARE
ELEVATED, SLIGHTLY DIFFERENT
KINETICS, FOR 4483 WE HAVE PEAK
AT ONE DAY AFTER CELL TRANSFER,
3061 PEAKED AT FOUR DAYS POST
CELL TRANSFER.
TNF SPIKES AT 50 TO 150
PICOGRAMS PER ML, AND THEN
CONTINUES ON ALMOST GOES BACK TO
BASELINE.
INTERFERON GAMMA SPIKES AT 150
FOR BOTH PATIENTS AND STAYS
ELEVATED FOR 11 DAYS AFTER CELL
TRANSFER.
AND SIMILAR GNCF RETURNED BACK
TO BASELINE RAPIDLY.
THE CYTOKINES TREATED BY
INTERFERON GAMMA, MAINLY BECAUSE
THE CYTOKINES SPIKED AT
APPRECIABLE LEVEL, 150 PICOGRAMS
PER ML, SUSTAINED, WE THINK
AFTER T CELL TRANSFER YOU WANT
TO GIVE T CELLS A GOOD SIGNAL
FOR A GOOD NUMBER OF DAYS AND
WITH ELEVATION IN CYTOKINES AT
LEAST 11 DAYS OUT.
SO, EXPLOITING INTERFERON GAMMA
PATHWAY TO ENHANCE T CELL-BASED
THERAPIES INTERFERON GAMMA
LEVELS ARE ELEVATED IN PATIENT
SERUM.
INTERFERON GAMMA IS PRODUCED
UPON TCR ENGAGEMENT, ANTIGEN
DEPENDENT, SELF-LIMITING.
YOU GET INTERFERON GAMMA
PRODUCTION AFTER IT'S GONE,
EVENTUALLY DECAYING OVER TIME.
INTERFERON GAMMA SIGNAL CAN BE
DETRIMENTAL, CAN LEAD TO T CELL
DEATH.
INTERFERON GAMMA MAY BE MORE
STABLE IN ACIDIC CONDITIONS
COMPARED TO INTERLEUKIN 2, NOT
ACTIVE AT PH 6 OR LESS.
THIS IS A SCHEMATIC OF THE
DESIGN OF THE CYTOKINE
RECEPTORS, A WILDTYPE, RECEPTOR
ON LEFT, EXISTS AS TWO
HETERODIMERS OF INTERFERON GAMMA
R 1, R 1 BINDS TO DIMER OF
INTERFERON GAMMA, EXPOSES THE
SITE THAT ALLOWS FOR BINDING OF
R 1 AND R 2 TO INITIATE
CROSS-PHOSPHORYLATION OF
RECEPTOR SUBUNITS RECRUITING
STAT1 SO INTERFERON GAMMA
INITIATES STAT1 SIGNAL.
INITIAL DESIGNS LOOK LIKE THIS.
SO FIRST IS CALLED CONVENTIONAL
CHIMERIC CYTOKINE RECEPTOR, WE
REPLACED TRANSMEMBRANE AND
CYTOPLASMIC DOMAINS OF WILD TYPE
INTERFERON GAMMA WITH RECEPTOR
OF CHOICE.
THE OTHER TYPE OF CHIMERIC
CYTOKINE RECEPTOR IS THE DOMAIN
SWATH CHIMERIC CYTOKINE
RECEPTORS, R 1 AND R 2 STAY WILD
TYPE EXCEPT FOR STAT1.
DETAILS THERE, WE DECIDED TO
LOOK AT STAT5 ACTIVATING
RECEPTORS AND SO AS WAS SHOWN
THIS MORNING, ALL SIGNS POINT TO
IL-2, WE THINK IT'S A POTENT T
CELL CYTOKINE, WE KNOW IT'S
POTENT.
WE TESTED INTERLEUKIN-2,
INTRACELLULAR DOMAINS, IL-3,
IL7, GM-CSF, AND FOR THE DOMAIN
SWAP CRRs WE PLACED STAT1
BINDING DOMAIN WITH IL-2
RECEPTOR STAT5 BINDING DOMAIN
FOR SIGNALING, TYROSINE 5110, WE
HAVE ANOTHER CONSTRUCT THAT HAS
THE SAME TYROSINE RESIDUE BUT
MUTATED SOX BINDING SITE,
SUPPRESSES CYTOKINE SIGNALING TO
HOPEFULLY ENHANCE STAT5
PHOSPHORYLATION.
WE REPLACED C-TERMINAL RESIDUE,
21 AMINO ACIDS OF THE
C-TERMINAL, IL-2 R BAITA GROWTH
HORMONE RECEPTOR SITES, AS WELL
AS CYTOPLASTIC DOMAINS, AND
ERYTHROPOEITIN RECEPTOR STITCHED
ON A BINDING SITE.
THIS IS OUR MODEL SYSTEM.
WE HAVE A TCR THAT RECOGNIZES
KRAS G12 EPITOPE.
CO-TRANSDUCE THAT WITH OUR
INTERFERON GAMMA R 1 R 2
CHIMERIC RECEPTOR, TWO VECTORS
HERE, CO-TRANSDUCED, C CELLS
THAT EXPRESS KRAS AND CHIMERIC
CYTOKINE RECEPTOR.
AND THEN WE EVALUATE EXPRESSION
OF THE CHIMERIC CYTOKINE
RECEPTOR, TRANSDUCTION
EFFICIENCY, TEST BIOCHEMICAL
FUNCTION BY PHOSPHO-STAT5 AND
TEST IN VITRO SERIAL KILLING
AGAINST TUMOR CELL LINES.
AND JUST HERE IS AN EXAMPLE OF A
PLOT SHOWING EXPRESSION AFTER
CO-TRANSDUCTION, TCRs HAVE
MOST CONSTANT REGIONS, ALPHA
BETA, WE CAN STAIN WITH MOUSE
ANTIBODY, THIS IS INTERFERON
GAMMA R 1, 20 TO 30%
TRANSDUCTION OF THE TCR AND
CHIMERIC RECEPTOR.
WHEN TESTING FOR BIOCHEMICAL
FUNCTION WE TAKE THESE CELLS,
WASH THEM, STARVE THEM OF
CYTOKINE OVERNIGHT AND STIMULATE
THEM WITH IL-2 OR INTERFERON
GAMMA.
WHAT WE HAVE HERE IS
TCR-ENGINEERED T CELLS ALONE.
STIMULATE WITH IL-2, YOU GET
NICE PHOSPHO-STAT5, WHEN YOU HIT
THEM WITH DOSES OF INTERFERON
DAM GAMMA WE DON'T GET
PHOSPHO-STAT5 BUT TEST TCR PLUS
INTERFERON GAMMA R 1 IL-2R BETA
CHIMERIC RECEPTOR GOOD STAT5,
YOU MIGHT RECALL THIS FROM THE
SERUM DATA, CERTAINLY ACHIEVABLE
IN PATIENTS AFTER CELL TRANSFER.
SO WHEN WE JUST SUMMARIZE ALL
THE OTHER CHIMERIC RECEPTORS
THAT I DESCRIBED, THEY ARE SHOWN
HERE, IL-3, IL-2, IL7, YOU CAN
SEE FOR THE MOST PART MOST OF
THE RECEPTORS FUNCTION
BIOCHEMICALLY TO ELICIT
PHOSPHO-STAT5, DMFI IS
DIFFERENT.
IL-2 AND GM-CSF INTRACELLULAR
DOMAINS ACHIEVE HIGHEST LEVELS
OF PHOSPHO-STAT5.
DOMAIN SWAP WE GET HIGH
FREQUENCIES OF PHOSPHO-STAT5,
BUT THE MFI IS LOWER THAN THE
CONVENTIONAL INTERFERON GAMMA
CHIMERIC RECEPTORS.
WE HAVE EVIDENCE THAT
FRANKENSTEIN RECEPTORS SEEM TO
WORK BIOCHEMICALLY, SO WE WANT
TO TEST THEIR FUNCTION.
DID THEY ENHANCE TCR T-CELL
THERAPY?
WE GOT ONLY 20 TO 30%
CO-TRANSDUCTION FACT SORTED THE
POPULATION OF DOUBLE POSITIVE
AND RAPIDLY EXPANDED THEM, OVER
90% OF CELLS EXPRESS BOTH TCR
AND INTERFERON GAMMA CHIMERIC
RECEPTOR, CO-CULTURE WITH H PAC
CELLS THAT NATURALLY EXPRESSES,
AND WE ENGINEERED THESE CELLS TO
EXPRESS GFP AS WELL.
ON DAY ZERO WE ADD 10,000
PANCREAS CANCER CELLS EVERY TWO
DAYS, IN A SERIAL KILL CAPACITY.
THIS GRAPH HERE IS REALLY THE
POSITIVE CONTROL, A TCR
ENGINEERED CELL, WITHOUT
CHIMERIC CYTOKINE RECEPTOR, IN
THE PRESENCE OR ABSENCE OF IL-2.
WITHOUT IL-2 YOU CAN SEE TCR IS
QUITE POTENT ON ITS OWN FOR THE
FIRST COUPLE KILLS.
STARTS TO LOSE ACTIVITY.
THIS IS GFP.
THE ENGINEERED CELLS CONTINUE TO
KILL OVER SIX OR SEVEN ROUNDS OF
TUMOR CELL WITH MORE IL-2.
RECEPTORS WILL LOOK LIKE THE
BLUE LINE HERE IN ABSENCE OF
IL-2.
SO THIS IS WHAT THE DATA LOOKS
LIKE WHEN WE DO THE SERIAL KILL
ASSAY.
THIS IS THE CHIMERIC RECEPTOR,
DOES IMPROVE SERIAL KILL.
CANCER CELLS.
SIMILAR WITH IL-3 RECEPTOR AND
GMPSF AS WELL.
SEEMS TO BE SOMEWHAT
RECAPITULATING ADDITION OF IL-2,
IN THE ABSENCE OF INTERLEUKIN-2
SO PRESUMABLY INTERFERON GAMMA
IS BEING MADE BY T CELLS, TAKEN
OUT AND SIGNALING THROUGH AN
IL-2 PATHWAY.
WHEN WE LOOK AT THE DOMAIN
SWAPPED CYTOKINE RECEPTORS,
INTERESTINGLY YOU CAN SEE THAT
EVERY SINGLE ONE OF THEM DOES
WORSE THAN THE TCR ALONE.
DESPITE HIGH FREQUENCY OF
PHOSPHO-STAT5, LOWER MFI TO BE
GRANTED, BUT THEY ALL DO WORSE
IN TCR SHOWN IN BLACK.
WE HAVE A HYPOTHESIS WHY THIS
MIGHT BE, WE'LL GET TO IN A
MINUTE.
THIS WAS I THINK UNEXPECTED
FINDING.
SO, WE WANTED TO DIVE DEEPER
INTO THE BIOLOGY OF THIS AND SO
WE WANTED TO TEST IMPACT OF THE
TRANSMEMBRANE DOMAIN, ALSO
INTRACELLULAR RECEPTOR PARTNER
ON FUNCTION OF THESE RECEPTORS.
AND SO THIS IS ORIGINAL DESIGN.
AGAIN TRANSMEMBRANE DOMAIN AND
CYTOPLASMIC DOMAIN FROM
INTRACELLULAR RECEPTOR.
WHAT WE DID WAS A TRANSMEMBRANE
DOMAIN SWAP SO NOW DERIVED FROM
INTERFERON GAMMA R1 AND R 2, AND
DURING SWAP AND INTRACELLULAR
PARTNER SWAP, LONGEST CHANGE ON
SHORTEST CHANGE AND VICE VERSA,
TWO ADDITIONAL CONSTRUCTS TO SEE
IF THAT WOULD ENHANCE OR IMPAIR
RECEPTORS.
SO I'VE SHOWN THIS DATA HERE.
I'M GOING TO GO THROUGH
INTERLEUKIN-2 AND ALSO GM-CSF,
ORIGINAL DESIGNS, WHEN WE DO THE
TRANSMEMBRANE SWAP, YOU CAN SEE
THAT NOW THEY LOSE ABILITY TO
CONTROL THE TUMOR CELLS.
SIMILARLY THEY LOSE ABILITY TO
CONTROL TUMOR CELLS.
ONCE YOU DO BOTH TRANSMEMBRANE
AND INTRACELLULAR PARTNER SWAP
IT DOES BETTER, SO THIS
PHENOTYPE NOW LOOKS MORE LIKE IF
YOU JUST ADDED IL-2.
FOR THE GMPCSF, THESE ARE THINS
YOU HAVE TO TEST EMPIRICALLY
WITH NO REAL PATTERN WOULD THIS
TRANSMEMBRANE INTRACELLULAR
DOUGH PLAIN TO USE.
WE CAN CONCLUDE THAT
TRANSMEMBRANE DOMAIN AND PARTNER
SWAB, INTRACELLULAR SWAP, CAN BE
IMPORTANT FOR FUNCTION.
DESPITE THIS ENCOURAGING DATA
SERIAL KILL DATA, THE CHIMERIC
RECEPTORS DON'T RECAPITULATE
PROLIFERATIVE SIGNAL OF IL-2
RECEPTOR.
WE TOOK TCR ENGINEERED CELLS AND
WITH CHIMERIC RECEPTOR, OUR BEST
CYTOKINE INTERFERON GAMMA
CHIMERIC RECEPTOR, EXPANDED IN
THE PRESENCE OF EITHER HIGH DOSE
IL-2 OR INTERFERON GAMMA.
WITH IL-2 ALONE, WITH TCR ALONE,
GOOD EXPANSION, NOT MUCH WITH
INTERFERON GAMMA.
HIGH DOSE INTERFERON GAMMA DOES
NOT GET TO LEVEL OF IL-2, MOST
OF THE TIMELESS THAN HALF FOLD
EXPANSION THAN IL-2, NOT QUITE
FULLY RECAPITULATING THE SIGNAL.
BACK TO THE HYPOTHESIS, THE ODD
FINDING THAT DOMAIN SWAP
RECEPTORS WEREN'T DOING WELL,
THIS IS OUR ONGOING HYPOTHESIS
WHY THE CHIMERIC CYTOKINE
RECEPTOR DOESN'T MEDIATE FULL
PROLIFERATIVE IL-2 SIGNAL.
T CELLS AT SOME POINT CAN
EXPRESS WILD TYPE INTERFERON
GAMMA RECEPTOR SHOWN HERE.
WE FORCE EXPRESSION OF THE
CHIMERIC RECEPTOR HERE, R 1 AND
R 32, POSSIBILITY OF MIXED DIMER
FORMATION.
THERE'S A POSSIBILITY THEY COULD
BIND AND MEDIATE SIGNALING
DETRIMENTAL TO THE T CELL.
SO THERE'S CERTAINLY A
POSSIBILITY THERE MAY BE MIXED
DIMER FORMATION HERE.
WITH THAT, THIS IS A WORK IN
PROGRESS BUT THOUGHT IT WAS AN
INTERESTING AND NEED WHY THE TO
SHARE.
WE KNOW TARGETING ONCOGENIC
DRIVER MUTATIONS CAN MEDIATE
REGRESSION OF METASTATIC CANCERS
BUT IMPROVEMENTS ARE NEEDED.
EN WE FOUND OUT THAT
TRANSMEMBRANE DOMAIN AND
INTRACELLULAR FUSION PARTNER IS
IMPORTANT.
LEAD CANDIDATE DOES NOT FULLY
RECAPITULATE IL-2 SIGNALING WE
THINK BECAUSE OF MIXED RECEPTOR
DIMERS.
WE'RE GOING TO KNOCK OUT TO SEE
IF THAT ENHANCES ACTIVITY OF
CHIMERIC CYTOKINE RECEPTORS, AND
PERFORM IN VITRO STRESS TESTING
OF THE CHIMERIC CYTOKINE
RECEPTORS USING ACIDIC
CONDITIONS, PRESENCE OF
TGF-BETA.
AND WE RECENTLY GOT IACUC
APPROVAL, WE'LL BE TESTING IN
MOUSE TUMOR MODELS.
I'M RUNNING OUT OF TIME.
FLASHING OUT GENERAL STRATEGIES
TO ENHANCE CELL THERAPIES, I
SHOWED ONE PROJECT.
I WANT TO HIGHLIGHT IN RED WE'RE
WORKING ON MULTIPLE FACETS ON
TRYING TO IMPROVE THIS THERAPY,
ESPECIALLY MANUFACTURED
CONDITIONS.
WE FOUND THIS MAKES A MUCH MORE
POTENT CELL, SO WE'RE MOVING
AHEAD WITH ADDING TGF-BETA TO
CELLS.
WE'RE TAKING A PAGE, HARNESS
FROM PERIPHERAL BLOOD.
OVEREXPRESSED TD 40, TO ENHANCE
T CELLS.
AND WE'RE WORKING ON NEXT
CLINICAL TRIAL, CONSIDERING ALL
THESE FACTORS TO COMBINE WITH
OUR TCR ENGINEERED T CELLS.
WITH THAT I'D LIKE TO
ACKNOWLEDGE A LOT OF PEOPLE
HERE.
ALL THE WORK I SHOWED WITH
CHIMERIC RECEPTOR DONE BY MY
POSTDOC, WHO IS AN EXTREMELY
DEDICATED AND HARD-WORKING
POSTDOC IN MY LAB.
I WANT TO ACKNOWLEDGE THE WHOLE
INSTITUTE MAINLY BECAUSE IT
TAKES A VILLAGE TO GET THE
PROGRAM UP AND RUNNING, THAT'S
WHY THERE'S SO MANY PEOPLE ON
THIS ACKNOWLEDGMENT SLIDE.
WALTER IS OUR DIRECTOR OF OUR
CANCER RESEARCH, AND HE'S BEEN
SUPER SUPPORTIVE OF OUR CELL
THERAPY EFFORTS.
SADLY HE WILL BE RETIRING AT TE
END OF THE YEAR, DR. ROSENBERG,
THANK YOU FOR EVERYTHING YOU'VE
DONE FOR THE PATIENTS, FOR THE
FIELD, AND PERSONALLY FOR MYSELF
BECAUSE YOU HELPED ME DEVELOP
INTO THE SCIENTIST I AM TODAY.
LAST AND NOT LEAST PATIENTS AND
THEIR FAMILIES, IT'S WHAT DRIVES
US AND IT'S OUR REASON FOR
EXISTING.
THANK YOU VERY MUCH.
HAPPY TO ANSWER ANY QUESTIONS.
[APPLAUSE]
>> YOU IF QUESTIONS FOR DR. TRAN
FIND HIM DURING THE BREAK.
RECONVENE BY 3:45, WE'LL ONLY BE
A FEW MINUTES BEHIND SCHEDULE.
PLEASE ENJOY THE BREAK.
FIRST THING YOU MIGHT NOTICE I'M
NOT RACHEL BYER WHO COULDN'T BE
HERE TODAY.
I HAVE THE HONOR OF MODERATING.
I'D LIKE TO BRING UP ONE OF THE
ALUMNI LEADERS THAT DR.
ROSENBERG MENTORS, THE PRESIDENT
APARTMENT OF MOFFITT,INE
MODIFIED T CELLS, IF
MICROINJECTION TO CRISPR
TECHNOLOGY.
>> THANK YOU.
IT'S A PRIVILEGE TO BE HERE,
WHAT AN HONOR.
THE 14 YEARS I SPENT NEAR
BETHESDA WITH DR. ROSENBERG, I
CHERISH MY WHOLE LIFE.
THERE'S NO BETTER PLACE ON THE
PLAN TO DEVELOP AS A PHYSICIAN
SCIENTIST THAN SURGERY BRANCH
THE NCI.
EVERY TIME I COME BACK IT FEELS
LIKE I'M COMING HOME.
WE DIDN'T JUST LEARN A LOT BUT
HAD FUN TOO, DIDN'T WE?
WE DID TRIATHLONS, WE HAD TRAVEL
SLIDES DURING SEMINARS, SKITS AT
CHRISTMASTIME, WE EVEN HAD A
SURGERY BRANCH ROCK BAND.
STEVE LABUDI WAS THE DRUMMER.
DO YOU KNOW WHAT WE WERE CALLED?
PROTOCOL VIOLATION.
VERY GOOD.
THAT WAS THE NAME OF OUR ROCK
BAND.
SURGERY BRANCH BAND WAS PROTOCOL
VIOLATION.
WHEN WE DID TRIALS WE HAD NO
VIOLATIONS.
WHAT WE STARTED WITH WAS NOT CAR
T BUT TNF.
WE'RE CYTOKINE PLACE, RIGHT?
TNF CAUSED TUMORS TO REGRESS,
WHEN WE PUT THE GENE IN IT
DIDN'T CRANK OUT SO MUCH TNF.
IT WAS A CHALLENGE, EARLY DAYS
OF GENE TRANSFER.
WE SAID LET'S JUST INJECT THE
CELLS ONE AT A TIME WITH A
MICROINJECTOR.
YOU HAD TO HIT THEM IN THE
NUCLEUS, IF YOU PUT TOO MUCH
VOLUME IN THEY WOULD EXPLODE.
IT'S ALSO WHERE WE STORED THE
MOUSE BEDDING, I CAME OUT
SMELLING LIKE MOUSE BEDDING.
IT WORKED BUT WASN'T REALLY THAT
SCALABLE AS YOU MIGHT IMAGINE.
WE GOT RETROVIRUSES TO WORK,
OPTIMIZING EVERYTHING.
WE HAD ALL OF THIS STUFF WE
COULD DO BY PUTTING GENES INTO T
CELLS AND ONE THING THAT WE
STARTED WITH, WORKING WITH
SOMEONE ON A SABBATICAL,
PRE-FIRST GENERATION, THIS WAS
OUR VECTOR.
WE HAD THREE ACTUALLY, ONE FOR
OVARIAN, BREAST, COLON.
THE ONLY ONE THAT WORKED, WE
DIDN'T KNOW WHY, OVARIAN.
WE WENT WITH IT.
AND THIS IS OUR FIRST PAPER,
1993, SHOWING THAT WE COULD GET
MELANOMA TIL TO RECOGNIZE AND
KILL OVARIAN CANCER WHEN WE PUT
IN CHIMERIC ALPHA RECEPTOR
AGAINST ALPHA FOLATE RECEPTOR.
THIS IS TIL 1143, AN INCREDIBLE
TIL LINE.
JIM YANG HELPED WITH IN VIVO
STUDIES, WE SHOWED WITH CANCER
MODELS WE COULD GET THIS TO WORK
IN VIVO.
AND WE COULD PUT THIS INTO BONE
MARROW GETTING THIS TO WORK IN
MICE, EVEN WHEN WE DEPLETED T
CELLS.
THIS RECEPTOR WORKED IN OTHER
EFFECTOR CELLS LIKE NK CELLS.
SO IT WAS A VERY EXCITING TIME.
WE THOUGHT LET'S TAKE IT TO THE
CLINIC.
WE DID TAKE IT TO A PHASE 1
STUDY IN OVARIAN CANCER, WHICH
WAS ALSO WE HAD A COMMENTARY IN
THE 20th ANNIVERSARY CCR ISSUE
ABOUT THIS FIRST CHIMERIC
ANTIGEN RECEPTOR TRIAL.
WE DIDN'T GET ANY RESPONDERS.
AT THE SAME TIME IT WAS SAFE, AN
EARLY STAGE, WE DIDN'T HAVE
CO-STIM MOLECULES AND WERE
GETTING NEUTRALIZING ANTIBODY
ISSUES.
WE GOT FIRST GENERATION, SECOND,
THIRD GENERATION VECTORS.
AND YOU'LL HEAR FROM THE NEXT
SPEAKERS WILL FDA APPROVAL AND
SUCCESSS FROM THE CAR PRODUCTS.
WHAT'S NEXT?
NOW THE TECHNOLOGY WENT FROM
MICRO INJECTION TO NOW YOU CAN
PRETTY MUCH TAKE OUT ANY GENE
YOU WANT IN A LYMPHOCYTE.
IT'S AMAZING.
YOU MAKE THE GUIDES.
WE'VE WORKED ON MANY DIFFERENT
GENES.
YOU CAN TAKE A GENE OUT WITH
CRISPR, PUT A GENE IN, REPLACE A
GENE, CHANGE THE GENE.
IT'S SO MUCH THAT YOU CAN DO
TODAY THAT WE COULDN'T DO BACK
THEN.
WE'RE HAVING A LOT OF FUN WITH
THAT.
THIS WAS SAM FIX IN THE LAB, YOU
CAN TAKE TGF-BETA RECEPTOR,
KNOCK IT OUT, YOU DO NOT SEE
PHOSPHO-MATTER.
WE PUBLISHED A REVIEW IN CANCER
DISCOVERY OF ALL THE DIFFERENT
WAYS YOU COULD USE CRISPR FOR T
CELL THERAPEUTICS.
YOU CAN MAKE THE CELLS LESS
TOXIC, TAKE OUT THE BREAKS, SO
IT'S AN EXCITING WORLD.
THIS IS WHAT WE DID OVER THE
LAST FEW YEARS.
WE SAID WHY DON'T WE CHANGE
METABOLISM OF T CELLS AND HELP
THE T CELLS PROLIFERATE AT TUMOR
SITE.
WE TOOK 300 METABOLISM GENES,
AGNOSTIC WAY, shRNA, PUT THEM
INTO THE CELLS DEVELOPED LEER,
AND INJECTED THEM INTO THE
ANIMALS AND TOOK OUT THE B 16
MELANOMAS AND SAID, ALL RIGHT,
WHAT KNOCKDOWNS ARE PRESENT
THERE?
IT'S VERY AGNOSTIC DOING THE
WHOLE SCREEN IN VIVO, A TALENTED
SCIENTIST WITH US, SORAYA.
A GENE ALLOWED US TO GET MORE T
CELLS AT THE TUMOR SITE WHEN WE
KNOCKED IT OUT.
WE DID THIS CONFIRMATION WITH
CRISPR.
THEN THE WORK BEGINS BECAUSE I
DON'T KNOW ALL 20,000 GENES AND
SO YOU'RE USING A LOT OF
WIKIPEDIA TO FIGURE OUT WHAT
THEY DO.
WITH THE PDHB COMPLEX, IT TAKES
YOUR PYRUVATE, CONVERTS TO
ACETYLCHOLATE, ALLOWS YOUR
GLUCOSE TO GO INTO OXIDATIVE
PHOSPHORYLATION PATHWAY, CAUSING
THE T CELLS TO GO INTO
GLYCOLYSIS BY BLOCKING IT,
OPPOSITE OF WHAT I THOUGHT WE
SHOULD HAVE DONE BUT IT'S WHAT
THE DATA SHOWED.
WE DID THIS TEN TIMES TO MAKE
SURE IT WAS RIGHT.
, BEING A SURGERY BRANCH.
THIS JUST SHOWED IN OUR 3D
MODEL, WE'RE USING TO USE 3D
MODELS MORE BECAUSE IT
RECAPITULATES WHAT WE SEE IN
VIVO MORE.
SO THIS IS -- LET'S ME JUST TAKE
THIS.
I'M GOING TO PLAY THIS VIDEO.
T CELLS ARE LABELED IN GREEN.
HERE THIS PURPLE BLOB ARE
TUMORS.
THIS IS THE KNOCKOUT CELLS.
THESE ARE THE NON-TARGETING
CONTROL.
YOU CAN SEE WHEN WE PUT T CELLS
TOGETHER WITH THE TUMOR YOU GET
HUGE PROLIFERATION OF THE T
CELLS WHEN YOU KNOCK OUT THE
PDHB GENE, I DON'T HAVE TIME TO
SHOW THE MECHANISMS.
WE'RE TAKING GLUCOSE, INSTEAD OF
OXIDATIVE PHOSPHORYLATION WE ARE
GOING TO AEROBIC GLYCOLYSIS,
EVEN THOUGH OXYGEN IS AROUND
WE'RE SENDING THEM THROUGH
GLYCOLYTIC PATHWAY, INCREASING
PENTOSE PHOSPATE PATHWAY,
OXIDATIVE AND NON-OXIDATIVE,
INCREASING LIPID SYNTHESIS, ALL
THINGS T CELLS NEED TO
PROLIFERATE.
SOUNDS FAMILIAR BECAUSE CANCER
CELLS DO THIS.
IT'S CALLED THE WARBURG EFFECT.
CANCER CELLS USE EVEN IN
PRESENCE OF OXYGEN UNDERGO
AEROBIC GLYCOLYSIS SO THEY CAN
MAKE ALL THESE SUBSTANCES,
IMPORTANT FOR THEM TO
PROLIFERATE.
I RECOLLECT NICK CALCULATED THE
NUMBERING OF LUNCHES WE HAD
TOGETHER, ALL OF US, IT WAS
BETWEEN THREE AND FOUR THOUSAND,
SO ONE OF THOSE LUNCHES I
REMEMBER STEVE WAS TALKING ABOUT
A DREAM EXPERIMENT WHERE MAYBE
THE T CELLS COULD BE LIKE
LEUKEMIA CELLS AND JUST
PROLIFERATE OUT, ALL OVER THE
TUMOR.
REMEMBER THAT?
AND JUST TAKE OVER THE TUMOR.
SO THIS IS A BIT LIKE THAT.
IT'S IN THAT DIRECTION WHERE
WE'RE GIVING THE T CELLS
TUMOR-LIKE CAPABILITIES TO
UNDERGO WARBURG EFFECT, AEROBIC
GLYCOLYSIS, SO THAT WE CAN --
THEY CAN MAKE ALL THE SUBSTANCES
THEY NEED TO GROW.
WHEN WE LOOK IN HUMAN DATABASE,
RESPONDERS VERSUS
NON-RESPONDERS, A WONDERFUL GUY
AT THE BROAD, WE SAW THAT
ACTUALLY IN SOME OF HIS
RESPONDER SUBSETS, IN GREEN
HERE, YOU CAN SEE THAT THE
CYTOTOXIC LYMPHOCYTES HAVE LOW
PDHB.
THAT'S ALL OF THE DATA I'M
SHOWING.
I WANTED TO TELL YOU ABOUT
MOFFITT.
WE'RE REALLY BIG ON CELL
THERAPY.
I DON'T KNOW IF WE SHOULD BE
CALLED SURGERY DOWN SOUTH OR
MOFFITT NORTH, JUST JOKING.
WHY ARE WE SO FOCUSED ON CELL
THERAPY, 1300 INFUSIONS ON THE
FLOOR?
WE INVITE YOU TO COME DOWN AND
VISIT US.
THE STORY IS THAT ONE OF OUR
CHAIRMAN OF THE BOARD WAS
SENATOR CONNIE MACK, REMEMBER
HIM?
HE WAS A SENATOR, PUSHED FOR
DOUBLING OF THE NIH BUDGET, HE
LOVED READING.
ONE DAY IN D.C. HE WAS EARLY FOR
A MOVIE WITH HIS WIFE PRISCILLA,
WALKS INTO A BOOK STORE, GUESS
WHAT I PICKS UP.
THE TRANSFORMED CELL.
HE PICKS UP THE TRANSFORMED
CELL.
HIS BROTHER DIED OF MELANOMA,
REALLY INTERESTED IN IT.
HE READ IT VORACIOUSLY, GOT HOLD
OF DR. ROSENBERG, BECAME GOOD
FRIENDS WITH HIM.
THEN WHEN HE BECAME CHAIRMAN OF
THE BOARD AT MOFFITT, HE SAID
WE'VE GOT TO DO THIS CELL
THERAPY AT MOFFITT, HOW DO WE DO
IT?
DR. ROSENBERG SAID RECRUIT JIM
YULET.
THEY WENT TO MICHIGAN, 2003, THE
REST IS HISTORY.
IT'S SO FOCUSED ON CELL THERAPY
BECAUSE OF THE TRANSFORMED CELL.
WE HAVE OUR OWN CELL THERAPY
PRODUCTION FACILITY.
WE EVEN STARTED GENE ENGINEERING
FACILITY SO WE CAN GET ALL OF
THE VIRAL VECTORS, MAKE THOSE
OURSELVES NOW.
WE DIDN'T BUY A MICROINJECTOR,
MAYBE THAT WILL BE ON THE LIST.
THEN WE ALSO -- PEOPLE SAY, OH,
CELL THERAPY, IT'S NOT SCALABLE.
THAT'S ABSOLUTELY FALSE.
IT IS 100% SCALABLE.
MAY NOT BE SCALED BY BIOLOGISTS
LIKE OURSELVES.
IT'S GOING TO BE SCALED BY
ENGINEERS.
THESE ENGINEERED ARE A DIFFERENT
SPECIES.
MATERIALS, THEY UNDERSTAND.
THIS WAS A VERY SPECIAL ENGINEER
RECRUITED TO START OUR
ENGINEERING PROGRAM, GREG
SAWYER.
HE HAD ADVANCED MELANOMA, HE
WENT ON A CLINICAL TRIAL AT
MOFFITT.
HE WAS WORKING ON THE MARS
ROVER.
CLINICAL TRIAL AT MOFFITT FOR
ADVANCE THE MELANOMA TEN YEARS
AGO.
JEFF WEBER PUT HIM ON A TRIAL OF
IMMUNOTHERAPY.
HE'S ALIVE, IMMEDIATELY STOPPED
WORKING WITH NASA, NOW HE'S
WORKING ON CANCER.
ONE OF HIS GOALS IS TO SCALE
CELL THERAPY.
EVEN WITH THE CAR Ts APPROVED,
ONLY 20% OF THE PATIENTS THAT
SHOULD BE GETTING THOSE ARE
GETTING THOSE.
ONE OF THE REASONS IS
SCALABILITY.
WE HAVE TO FOCUS ON THIS.
IT'S GOING TO BE THROUGH
ENGINEERS.
WORKING WITH ENGINEERS.
SO WE'RE HAPPY.
THIS IS I WANTED TO THANK OUR
LAB.
I WANT TO MAKE A TRIBUTE TO DEB.
OUR MD/PHD STUDENT PASSED AWAY A
COUPLE YEARS AGO.
I THANK THE NCI SURGERY GROUP
FOR GIVING HER A NOVEL T-CELL
RECEPTOR, THE BEST CHANCE
POSSIBLE.
SO NOT ONLY IS THIS A GREAT
PLACE, IT'S OUR HOME.
IT'S A PLACE WHERE INCREDIBLE
COMPASSIONATE CARE IS GIVEN, SO
THE CLINICAL SERVICE HERE IS
LIKE NO OTHER.
WE REALLY APPRECIATE YOUR HELP
WITH DEB.
JUST SHOWS HOW MUCH MORE WE HAVE
TO DO.
I WANT TO TELL YOU THROUGH MY
LEADERSHIP JOURNEY WHAT WOULD
STEVE ABOUT, I THINK.
WHEN I GET IN A BIND, IT'S WHAT
I THINK.
MANY QUOTES.
PUSHING THE ENVELOPE.
HE JUST SAID KEEP DOING THAT.
BEING PATIENT.
IF YOU KEEP THE PATIENT'S BEST
INTEREST AT HEART IT WILL ALWAYS
BE THE RIGHT DECISION.
WHEN I GET IN A BIND I THINK
WHAT WOULD STEVE DO?
HE'S ALWAYS IN MY HEAD.
I WANT TO END, MY INVOLVEMENT,
WE WERE VERY EXCITED TO START
THE ROSENBERG FUND, YOU HEARD
FROM ONE OF OUR FIRST
RECIPIENTS.
AND STEVE HAS DONE SO MUCH FOR
YOUNG FACULTY, WHAT BETTER
PERSON, THERE'S NO BETTER PERSON
IN THE UNIVERSE TO REPRESENT A
SCHOLARSHIP FOR YOUNG FACULTY.
EVERY YEAR WE GET 50 TO 100
APPLICATIONS, FUND ONE
INCREDIBLY FORTUNATE PERSON.
IT CHANGES THEIR LIFE.
WE'RE CONTINUING TO TRY TO RAISE
MORE MONEY TO HELP MORE PEOPLE
AND HERE IS THE CODE, IF YOU OR
YOUR COMPANY WANTS TO FURTHER
ENDOW THIS AWARD THAT'S GOING TO
GO ON FOREVER, WELL BEYOND US,
SO IT WILL GO ON FOR FOREVER.
REBECCA GREGORY IS OUR HEAD OF
FUNDRAISING AT SITC, SHE'S RIGHT
THERE, TALK TO HER IF YOU HAVE
IDEAS HOW TO FURTHER ENDOW THIS
INCREDIBLY IMPORTANT PROGRAM FOR
THE ROSENBERG SCHOLARSHIP.
THANKS FOR YOUR TIME, IT'S SUCH
AN HONOR TO BE HERE.
[APPLAUSE]
>> THANK YOU FOR THE LOOK BACK
AND ACTUALLY INJECTING INTO THE
NUCLEUS OF THE CELL, SOMETHING I
DON'T THINK ANYBODY HAS THOUGHT
ABOUT FOR A WHILE, FORTUNATELY
FOR ME.
I'D LIKE TO WELCOME OUR NEXT
SPEAKER, JIM KOCHENDERFER SPENT
TIME IN THE ROSENBERG LAB, SINCE
DEVELOPED A SUCCESSFUL LAB AND
CLINICAL OPPORTUNITIES OF HIS
OWN WHICH LED TO DEVELOPMENT OF
CD19 FOR PATIENTS WITH LYMPHOMA.
PLEASE WELCOME JIM TO THE STAGE.
>> I'D LIKE TO THANK THE
ORGANIZERS FOR INVITING ME TO
TALK TODAY.
I'M FROM SENIOR INVESTIGATOR AND
SURGERY BRANCH, CONTINUING TO
WORK WITH DR. ROSENBERG,
PRIVILEGED AND THANKFUL TO DO
SO.
I'M GOING TO TALK ABOUT
HEMATOLOGIC
HEMATOLOGIC MALIGNANCIES, IT'S
ALL I DO.
I STARTED WORKING WITH STEVE IN
MARCH OF 2007, THAT WAS LIKE
DEFINITELY THE BEST PROFESSIONAL
DECISION OF MY LIFE.
I WAS THE ONLY PERSON WORKING ON
HEMATOLOGIC MALIGNANCIES, STEVE
SUGGESTED THAT I WORK ON THIS
ANTI-CD19 CAR T CELL PROJECT.
I WAS GOING TO WORK ON SOME TYPE
OF HEMATOLOGIC PROJECT, THAT
SUGGESTION POINTS OUT LIKE ONE
OF THE MOST IMPORTANT ATTRIBUTES
HE HAS WHICH IS ABILITY TO PICK
THE RIGHT PROJECT TO WORK ON.
THE MOST IMPORTANT THING HE SAID
IS PICKING THE RIGHT PROJECT TO
WORK ON, SO THANKFUL HE'S SO
GOOD AT PICKING THE RIGHT
PROJECT BECAUSE THAT HELPED OUT
MY CAREER IMMENSELY.
MORE THAN ONCE.
SO, ANOTHER POINT BACK IN THE
OLD DAYS OF 2007, IT WAS SUCH A
SMALL FIELD BACK THEN, ONLY A
FEW LABS ACROSS THE COUNTRY
DOING ADOPTIVE T-CELL THERAPY
FOR CANCER.
AND THERE WAS MINIMAL INDUSTRY
INVOLVEMENT SPECIFICALLY WITH
CELL THERAPY AT THAT TIME.
AND I THINK THE FIRST PART OF MY
TALK WILL SHOW HOW THAT CHANGED
SO MUCH, IN A SHORT PERIOD OF
TIME.
SO, THIS ANTI-CD19 CAR PROJECT
WE WORKED ON AS I MENTIONED
STARTED IN 2007.
I DESIGNED FMC 6328 DIS
PUBLISHED IN JOURNAL OF
IMMUNOTHERAPY IN 2009.
AND ALMOST IMMEDIATELY STARTED A
CLINICAL TRIAL ONE OF THE
STRENGTHS OF THE NCI TO
TRANSLATE THINGS INTO CLINICAL
TRIALS.
TRIAL WAS SUCCESSFUL, THE FIRST
PERSON, THAT'S THE CT SCANNER,
MASS IN CHEST, PLEURAL EFFUSION,
18 WEEKS LATER GONE, PLEURAL
EFFUSION IS GONE, ZERO TOXICITY,
AN AMAZING FIRST PATIENT.
EVEN MORE AMAZING THAN CLINICAL
RESPONSE IF YOU CAN SEE THE FLOW
PLOT THERE, MAINLY FOCUSING ON
CD19 ON THE Y-AXIS, BEFORE CAR T
CELL INFUSION HAD CELLS IN HIS
BLOOD, UPPER LEFT QUAD RANT,
THAT'S BONE MARROW, NOT BLOOD,
36 WEEKS LATER NO CD19 POSITIVE
CELLS AT ALL IN HIS BODY, I WENT
TO SHOW DR. ROSENBERG THE PLOTS,
INCREDIBLE TO ME THIS WORKED SO
WELL THAT WE ERADICATED EVERY
LAST B CELL IN THIS PATIENT,
BLOOD, BONE MARROW, COULDN'T
FIND ANY B CELLS, INCREDIBLE.
I DID MOUSE STUDIES THAT
REPLICATED THE SAME THING.
HE SAID WE DID MOUSE STUDIES
FIRST, THEN HUMAN, I WAS
ACTUALLY WORKING ON BOTH AT THE
SAME TIME.
WE SHOWED THAT.
AND THEN OVER THE YEARS THERE WE
WERE MAKING RAPID PROGRESS,
ITERATIONS ON THIS ANTI-CD19 CAR
T CELL APPROACH, THIS WAS OUR
FINAL ITERATION IN SURGERY
BRANCH USING LOW DOSE
FLUDARABINE AND
CYCLOPHOSPHAMIDE, THE
CONDITIONING REGIMEN WE SETTLED
HERE, THAT'S COMMONLY USED NOW
BY PRETTY MUCH ALL COMMERCIAL
CAR PRODUCTS, FIRST DONE IN
SURGERY BRANCH IN THE SETTING OF
CAR-T CELLS.
IN OUR FINAL ITERATION IN A
PAPER IN JOURNAL OF CLINICAL
ONCOLOGY 2017, 45% OF PATIENTS
HAD DURATIONS OF RESPONSE
GREATER THAN TWO YEARS, 45% OF
THE TOTAL PATIENTS IN THE TRIAL
HAD DURATION OF RESPONSE GREATER
THAN TWO YEARS.
JUST A COUPLE EXAMPLES OF THESE
PATIENTS, THE LARGE BURDEN OF
THE DISEASE THAT COULD BE
ELIMINATED, THE RED -- OOPS.
I'M NOT SURE HOW TO DO THE
POINTER.
RED AREA IS LYMPHOMA, A LARGE
MASS IS GONE TEN MONTHS LATER.
THE HEART AND KIDNEYS STILL RED
BECAUSE THEY ARE AT THE PLACE
WHERE THE CONTRAST ACCUMULATES.
BUT THE PATIENT WAS IN COMPLETE
REMISSION, SHOWN BELOW THE
YELLOW PART IS LYMPHOMA,
COMPLETELY GONE.
ONE OF MANY PATIENTS IN THE
TRIAL WITH GREAT OUTCOME.
2020 PUBLISHED A PAPER WITH
KATIE, A FELLOW WORKING WITH ME,
LONG-TERM FOLLOW-UP FROM THIS
TRIAL, MEDIAN EVENT FREE
SURVIVAL 55 MONTHS, 51% OF THE
PATIENTS HAD RESPONSES GREATER
THAN 3 YEARS, AND LONGEST
SURVIVING PATIENT, PICTURE IN
YOUR PROGRAM WITH DR. ROSENBERG,
IS STILL IN COMPLETE REMISSION,
HE'S BEEN IN EVENT-FREE SURVIVAL
FOR 14 YEARS, SO THAT'S THE
LONGEST CONTINUING SURVIVING CAR
T PATIENT IN THE WORLD.
YOU CAN SEE THE GRAPH WITH
EVENT-FREE SURVIVALS, B CELL
MALIGNANCIES, LOW GRADE LIMB
FOAMA CLL, WE TREATED ALL THREE
TYPES.
ALSO THE GRAPH ON THE RIGHT-HAND
SIDE SHOWING THE PLATEAU IN
EVENT-FREE SURVIVAL IN LARGE
B-CELL LYMPHOMA PATIENTS THAT
ATTAIN C.R.
HOW THE FIELD HAS CHANGED WE NOW
HAVE FOUR FDA-APPROVED PRODUCTS.
I WON'T READ OUT THE NAMES
THERE.
IT'S BASICALLY THE EXACT PRODUCT
WE USED IN SURGERY BRANCH, FINAL
ITERATION, A LOT OF PEOPLE WENT
INTO, YOU KNOW, MAKING THAT
PRODUCT, MAKING THAT A
COMMERCIAL PRODUCT BUT IN
SURGERY BRANCH PEOPLE THAT WENT
INTO THAT, CERTAINLY MARK DUDLEY
AND STEVE FELDMAN WERE
INSTRUMENTAL, RICK MORGAN WAS A
HUGE HELP, GENETIC ENGINEERING,
MOLECULAR BIOLOGY.
IT COMES DOWN TO TWO CAR T CELL
PRODUCTS, BOTH USE THE SAME CAR
WE MADE IN SURGERY BRANCH.
FOUR CAR T CELL PRODUCTS AND I
JUST CHECKED THIS WEEK BECAUSE I
THOUGHT IT WOULD BE INTERESTING,
OF THE CAR THAT WAS MADE IN
SURGERY BRANCH BACK IN 2007,
KITE TREATED GREATER THAN 23,000
PATIENTS, THAT'S AS OF TWO DAYS
AGO, I ASKED THEM.
THAT'S UP TO DATE NUMBER.
YOU CAN SEE HOW MUCH IT GREW.
WE HAVE FDA-APPROVED CARS FOR
LYMPHOMA INDICATIONS, ACUTE
LYMPHOID LEUKEMIA, CHRONIC
LYMPHOCYTIC LEUKEMIA, MULTIPLE
MYELOMA TWO CAR T CELL PRODUCTS
FDA APPROVED.
THAT WAS THE HISTORICAL PART OF
THE TALK.
NOW I'M GOING TO TALK ABOUT SOME
OF STUFF GOING ON MY LAB NOW.
I HAVE A LAB IN SURGERY BRANCH.
WE STILL DO ALL CELL THERAPY FOR
HEMATOLOGIC MALIGNANCY.
THE FIRST IS FOR GAMMADELTA T
CELL MALIGNANCIES, EXTREMELY
RARE TYPE OF MALIGNANCY.
IT HAS A HETERODIMER, THEY MAKE
UP 1 TO 10% OF BLOOD T CELLS.
MOST OF THE EXPRESS B DELTA 2
RECEPTOR, THE ROLE IS NOT CLEAR.
THERE'S EVIDENCE THEY ARE
IMPORTANT FOR IMMUNITY,
ESPECIALLY AGAINST CERTAIN
PATHOGENS, MYCOBACTERIA,
TUBERCULOSIS.
THERE'S NO IMMUNODEFICIENCY
DISEASE CHARACTERIZED BY
DEFICIENCY, WE CAN'T SAY WHAT
HAPPENS IF A PATIENT LACKS ALL
THEIR GAMMADELTA T CELLS.
THESE ARE VERY RARE
MALIGNANCIES, WHICH IS ACTUALLY
SOMETIMES CONSIDERED A PLUS WHEN
YOU'RE AN NIH INVESTIGATOR
BECAUSE IT'S SOMETHING THAT
WE'RE ENCOURAGED TO LEARN TO
TREAT VERY RARE DISEASES THAT
INDUSTRY MIGHT NOT HAVE AN
INTEREST IN AND BIGGER GROUPS
MIGHT NOT HAVE INTEREST IN.
SO, YOU CAN SEE WE HAVE
HEPATOSPLENIC T CELL LYMPHOMA,
FIVE YEAR OVERALL SURVIVAL 10 TO
20%.
T CELL ACUTE LYMPHOPLASTIC
LEUKEMIA, ANOTHER SERIOUS TYPE
OF LEUKEMIA, AND THEN VERY RARE
INTESTINAL LYMPHOMA, 7 TO 7.5
MONTHS, DISEASES I HELP TO
TREAT -- LEADERSHIP TO TREAT
WITH A GAMMADELTA CAR.
WE CONSTRUCTED A SERIES OF
CARs, TO SHOW SOME OF THE
EFFORT INTO MAKING CARs WE
ASSESS TEN ANTIGEN RECOGNITION
DOMAINS AND SELECT THE TWO BEST
FOR EVALUATION.
A POSTDOC IN MY LAB MADE 23 TO
COME UP WITH FACTORS NOW IN THE
FINAL STAGES OF EVALUATING WHICH
I'LL TALK ABOUT IN THE NEXT FEW
SLIDES.
WE TESTED VARIOUS PARTS OF THE
CARs, SOME DIAGRAMS OF CARs
WE TESTED, ONE FACTOR THAT I
HAVE FOUND RECENTLY WHICH CAN BE
AN IMPORTANT FACTOR IN CAR
FUNCTION IS LINKER, CARs HAVE
A SINGLE CHAIN FV, HEAVY CHAIN
RARABLE DOMAIN, LIGHT CHAIN OF
ANTIBODY, JOINED BY LINKER, AND
THE ONE WE USE IN MY CAREER HAS
BEEN 218 LINKER, WHITTLOW
LINKER.
WE'VE BEEN TESTING A LONGER
VERSION OF A G 4S LINKER.
LINKING THAT CAN SOMETIMES HELP
WITH CAR FUNCTION.
PERHAPS HAVING FUNCTIONAL
ADVANTAGES.
AND WE ALSO CAREFULLY LOOKED AT
THE TRANSMEMBRANE DOMAIN,
SOMETIMES DOESN'T GET THE MOST
ATTENTION IN CARs.
BUT WE FOUND TO BE VERY
IMPORTANT.
SO THE CARs ON THIS PAGE ONE
HAS A LONGER LINKER, AND 228
CARs, SO, AGAIN, WE WON'T TALK
ABOUT ALL 23 CARs BUT JUST
THOSE FOUR, AND THE PROCESS WE
GO THROUGH FIRST TO DO
EXPRESSION ANALYSIS TO SEE IF
THE CAR IS EXPRESSED ON THE T
CELLS, IN VITRO FUNCTIONAL
TESTING USING INTERFERON GAMMA,
ELISAs TO START WITH, AND
WE'RE EVALUATING CD28-CONTAINING
CARs TO SINGLE CHAIN, FOR FOUR
CARs, AND RED BAR SHOWING THE
ONE WE FOUND OUT TO HAVE THE
HIGHEST DEGREE OF SPECIFIC
RECOGNITION OF OUR TARGET CELLS,
WHICH WAS MGDL 28D AGAINST THREE
GAMMADELTA LYMPHOMA T CELL
LINES, WITH NEGATIVE CONTROL
CELL K 562, MINIMAL INTERFERON
GAMMA RELEASED THERE, T CELLS
ALONE.
THIS WAS ALL STANDARD ELISA, TO
DETERMINE THE CAR WHICH
RECOGNIZED MOST STRONGLY THE
LYMPHOMA CELLS.
WE'VE STARTED TESTING IN MICE,
USING NSG MICE, HIGHLY
IMMUNOCOMPROMISED MICE THAT GET
A SINGLE TREATMENT OF SINGLE
INJECTION OF CAR-T CELLS AFTER
THE TUMOR CELLS ARE ESTABLISHED.
DAY ZERO, MALIGNANCY ESTABLISHED
IN MICE.
THEY RECEIVE INJECTION OF T
CELLS ON DAY ZERO.
THEY DAYS LATER CANCER STARTING
TO DROP,.
WE HAVE A DOSE-DEPENDENT
REJECTION OF THE TUMOR.
THIS WAS A NICE OUTCOME.
WE'RE LEANING TOWARD THAT CAR.
HAVEN'T DECIDED WHICH TO GO
FORWARD YET, TESTING A COUPLE,
HAVEN'T DECIDED WHAT HINGE
REGION TO USE.
BUT THE OVERALL GOAL IS BECAUSE
THESE GAMMADELTA T CELL
MALIGNANCIES ARE RARE WITH BAD
PROGNOSIS, TO CONDUCT A CLINICAL
TRIAL, WE'RE PLANNING ON
STARTING A SMALL PHASE 3 TRIAL
ONCE WE GET OPTIMAL CAR DETECTED
AND FINISH MORE MOUSE
EXPERIMENTS, GET REGULATORY
APPROVALS, HOPEFULLY IN THE NEXT
COUPLE YEARS WILL START A TRIAL
TESTING ONE OF THESE CARs
AGAINST GAMMADELTA T CELL
MALIGNANCIES.
FINAL SEGMENT, THIS IS COMING
BACK TO A LOT OF PREVIOUS TALKS
FOR THE DAY, PEOPLE IN THE
BRANCH WORKING WITH NEOEPITOPE
SPECIFIC T-CELL RECEPTORS, AND
AGAIN I'M UTILIZING EXPERTISE OF
MY COLLEAGUES TO HELP APPLY
THINGS THEY HAVE DONE IN SOLID
TUMORS TO HEMATOLOGIC
MALIGNANCIES.
SOME TCRs HAVE BEEN COVERED TO
STREET HEMATOLOGIC MALIGNANCIES.
THE REASON I WANT TO DO THIS
CAR-T CELLS ARE EFFECTIVE MORE B
CELL, BUT IDEAL TARGETS FOR
OTHER MALIGNANCIES ARE HARDER O
IDENTIFY.
WE KNOW THAT AML AND
MYELODYSPLASTIC SYNDROME ARE
SUSCEPTIBLE BECAUSE THEY HAVE
BEEN TREATED WITH ALLOGENEIC
TRANSPLANT SINCE THE 70s, IT
HAS SIDE EFFECTS,
GRAFT-VERSUS-HOST DISEASE CAN BE
DEADLY.
WE HOPE TO ENHANCE OR REPLACE
ALLOGENEIC TRANSPLANT.
THAT'S THE GOAL.
WE'RE A LONG WAY FOR AML, BUT AT
LEAST WE CAN MAYBE ENHANCE IT OR
REPLACE IT IN SOME CASES.
UNLIKE CAR-T CELLS, ANOTHER
ADVANTAGE THEY CAN TARGET
INTRACELLULAR ANTIGENS.
SO THE NEOANTIGEN SOMATIC
MUTATIONS ARISE IN TUMOR CELLS,
SOMETIMES SOMATIC MUTATIONS TO
GIVE RISE TO NEW EPITOPES,
RECOGNIZED BY TCRs, THIS HAS
BEEN A TARGET OF INTENSIVE
EFFORTS IN THERAPY BRANCH FOR A
LONG TIME.
AND TWO OF THE NEOANTIGENS ARE
TP 53 AND RAS, I SAY RAS TO MEAN
KRAS, H-RAS, N-RAS.
I PUT THEM ALL TOGETHER TO SAVE
TIME.
SO WE'RE TESTING A VARIETY OF
TCRs, WHICH ARE HERE IN THIS
TABLE, A TOTAL OF EIGHT WE'RE
EVALUATING FOR USE WITH
HEMATOLOGIC MALIGNANCIES.
THE FIRST THREE ARE TARGETING
TP53 NEOEPITOPES, THEN WE HAVE
FIVE RAS EPITOPES.
SO SEVERAL SURGERY BRANCH
INVESTIGATORS DISCOVERED THESE
TCRs, AND THE LIBRARIES, MUCH
OF THE WORK DONE BY LEVIN AND
KIM IN PUBLICATIONS AT THE
BOTTOM OF THE SLIDE.
SO WE'VE BEEN TESTING THESE TO
EVALUATE THEIR ABILITY TO SHOW
RECOGNITION OF HEMATOLOGIC
MALIGNANCIES, ALSO TO CONFIRM
THE FINE SPECIFICITY.
THIS SLIDE IS SHOWING AN
EXPERIMENT WHERE WE TRANSDUCE
CELLS WITH HLA FOR ONE OF THE
NEOEPITOPES, TRANSFECT WITH GENE
ENCODING THE NEOEPITOPE OR WILD
TYPE GENE TO SHOW FINE
SPECIFICITY OF THE TCRs.
YOU CAN SEE THAT INTERFERON GAM
IS ON THE Y-AXIS.
THE BLUE BAR INDICATES A LOT OF
INTERFERON GAMMA RELEASE WITH
THE RIGHT HLA TYPE, A-1, WITH
THE RIGHT MUTATION, G12B RAS IN
THE SAME CELL.
IF WE LACK COMPONENTS SUCH AS
LACKING THE RIGHT MUTATION OR
LACKING THE RIGHT HLA TYPE,
THERE'S MINIMAL INTERFERON GAMMA
RELEASE.
AGAIN, THIS IS LIKE THE KEY, WE
NEED SPECIFICITY.
OF ALL THE EIGHT TCRs THE
TABLE ON THE RIGHT SHOWS RATIO
OF INTERFERON GAMMA RELEASE WHEN
THE COS CELL WAS TRANSFECTED,
YOU CAN SEE THERE'S A LARGE
RATIO OF INTERFERON GAMMA
RELEASE WITH MUTATED VERSUS WILD
TYPE WITH HIGHER INTERFERON
GAMMA BEING WITH MUTATED GENE IN
THE CELL.
AND, AGAIN, SUBSTANTIAL
INTERFERON GAMMA RELEASED IN ALL
THESE EXPERIMENTS, ALL TCRs
WORKED WONDERFULLY AS EXPECTED.
WE'VE GONE ON TO START TESTING
AGAINST LEUKEMIA IN MICE,
MULTIPLE MYELOMA.
THIS SHOWS MULTIPLE MYELOMA, NSG
MICE, STARTED A TUMOR CELL LINE
KMS 26 LUCIFERASE, AFTER
ESTABLISHED DAY ZERO AGAIN
ESTABLISHED TUMOR BURDEN, WE
HAVE UNTREATED MICE, IRRELEVANT,
AND MICE THAT REACHED TCR CELLS.
MICE THAT RECEIVED THE CORRECT T
CELLS, R 175H, A GOOD T-CELL
RECEPTOR HAD CLEARANCE OF THEIR
MULTIPLE MYELOMA, AND ALL THE
WAY DOWN TO THE END OF THE
EXPERIMENT WE DIDN'T SEE
RELAPSE, NOW USED WITH MULTIPLE
MYELOMA AND AML CELL LINE, WORKS
AGAINST BOTH HEMATOLOGIC
MALIGNANCIES, SURVIVAL IN
AGREEMENT WITH MOUSE IMAGES.
WITH THE NEOEPITOPE PROJECT WE
HAVE T CELLS SPECIFICALLY
RECOGNIZING CORRECT NEOEPITOPES,
ELIMINATING TUMORS IN MICE, WE
HELP TO START A CLINICAL TRIAL
MAKING USE AGAIN OF OUR GREAT
COLLABORATION WITH OUR
COLLEAGUES IN SURGERY BRANCH
WHETHER WE HAVE USABLE T CELL
TCR VECTORS AVAILABLE TO USE IN
A CLINICAL TRIAL.
WE'RE WELL ALONG THE WAY OF
GETTING REGULATORY APPROVAL,
HOPEFULLY TREATING PATIENTS
EARLY IN 2025.
I'D LIKE TO JUST FINISH UP,
THANKING DR. ROSENBERG AND
CO-WORKERS, LAUREN CUTMORE,
KATHERINE CAP, WHO DID THE IN
VITRO WORK, OTHER COLLEAGUES IN
MY LAB, AND OTHERS.
AGAIN, THANKS TO EVERYBODY.
[APPLAUSE]
>> THANK YOU, JIM.
WE'VE COME TO THE LAST TALK OF
THE DAY.
THANK YOU FOR STICKING WITH US.
I'M EXCITED TO HEAR DR. MACKALL
SPEAK BECAUSE WE'RE SEEING
EVIDENCE CAR IN SOLID CANCERS IS
HAVING CLINICAL BENEFIT AND NOW
MORE OBJECTIVE RESPONSES, AND
ALWAYS HAPPY TO HEAR HER TALK
ABOUT HER WORK.
PLEASE WELCOME DR. MACKALL TO
THE STAGE.
[APPLAUSE]
>> THANK YOU.
I MIGHT BE THE ONLY SPEAKER
TODAY NOT A MEMBER OF THE
SURGERY BRANCH.
AND I HOPE THAT MAYBE I COULD BE
CONSIDERED AN HONORARY MEMBER
BECAUSE IN FACT I HAVE THE
SIMILAR SLIDE THAT YOU'VE SEEN
TODAY, IN 1985 I WAS AN INTERN
IN AKRON, OHIO, HAVING GRADUATED
FROM A LOCAL MEDICAL SCHOOL
THERE, WITH AN INTEREST IN
ONCOLOGY, DOING AN INTERNAL
MEDICINE PEDIATRICS RESIDENCY.
I KNEW I WANTED TO DO ONCOLOGY.
I HADN'T DECIDED PEDIATRIC OR
ADULT.
I REMEMBER THIS LIKE YESTERDAY,
I LEARNED TODAY THAT THE PATIENT
WHO RESPONDED IN THIS PAPER I
GUESS IS WITH US TODAY, IT'S
JUST AMAZING TO SEE IT COME FULL
CIRCLE.
I SAID THAT'S WHAT I WANT TO DO.
WHERE WAS IT BEING DONE?
AT THE NIH.
IN 1989, I MOVED FROM OHIO TO
THE NIH, AND WAS HERE FOR 27
YEARS.
I WAS NEVER ON THE THIRD FLOOR.
I WAS FIRST ON THE 13th FLOOR,
THEN FIRST FLOOR, BUT DECIDED TO
PURSUE PEDIATRICS, BUT WITH THE
GOAL OF DEVELOPING IMMUNOTHERAPY
FOR CHILDREN'S CANCERS.
I DIDN'T HAVE ROLE MODELS IN
THAT SPACE BUT HAD STEVE
ROSENBERG AS A ROLE MODEL.
AND HE REALLY TAUGHT ME WHAT
NEEDED TO BE DONE.
THE NIH INTRAMURAL PROGRAM
SHOWED ME HOW TO TAKE IDEAS FROM
BENCH TO BEDSIDE.
I'M GOING TO TALK AND GET
EVERYBODY OUT ON TIME, ABOUT THE
PROSPECT OF USING CAR-T CELLS
FOR SOLID TUMORS.
AND I KNOW THERE MAY BE SOME
AMONG US WHO THINK THAT WILL
NEVER WORK BECAUSE, YES, THERE
ARE NO TRULY TUMOR-SPECIFIC CELL
SURFACE MOLECULES.
BUT IT IS MY BELIEF AND I THINK
BACKED UP BY DATA THAT THERE ARE
SEVERAL MOLECULES FOR WHICH
THERE IS A POTENTIAL THERAPEUTIC
WINDOW WHERE WE CAN THREAD IT
BASED UPON THE RELATIVELY HIGHER
ANTIGEN DENSITY THAT CAR-T CELLS
NEED TO FIRE AND THEREFORE THEY
WILL IGNORE NORMAL TISSUES.
WE ALL KNOW THAT THE CAR T CELL
RESULTS IN SOLID TUMORS HAVE
BEEN NOTHING LIKE WHAT WE'VE
SEEN IN LIQUID TUMORS BUT I
WOULD ARGUE THAT IN THE LAST FEW
YEARS WE HAVE STARTED TO SEE
SIGNALS OF ACTIVITY.
AND THIS IS A LIST OF SOME OF
THOSE.
YOU'LL NOTE THAT QUITE A FEW ARE
HAPPENING IN PEDIATRICS.
AND UNLIKE ADULT CANCERS WE'VE
HEARD SO MUCH TODAY ABOUT
MUTATED NEOANTIGENS, ABILITY TO
GO AFTER MOTHER NATURE'S KEY
CELL, TO RECOGNIZE THESE NEW
ANTIGENS BUT TUMORS IN CHILDREN
ARE TYPICALLY VERY, VERY
[ LOWLY
MUTATED.
WE HAVE TO PARTICULAR THE IMMUNE
SYSTEM INTO RESPONDING TO
TUMORS.
I'LL TALK ABOUT ONE STORY USING
A CAR TARGETING GD2 GANGLIACIDE,
IN DIFFUSE MIDLINE GLIOMA DRIVEN
BY MUTATION IN THE H3 HISTONE,
THAT PREVENTS TRIMETHYLATION,
UNABLE TO TURN OFF GENES.
IT LEADS TO WIDESPREAD
TRANSCRIPTIONAL DYSREGULATION
AND ONE OF THE GENES THAT GETS
TURNED ON ARE THE GENES THAT
DRIVE THE PRODUCTION OF THIS GD2
GANG LEOCIDE.
WHAT WE'VE DISCOVERED IN THE
LABORATORY OF A PIONEERING
NEURAL ONCOLOGIST AT STANFORD,
TUMORS THAT ARE NOT DRIVEN BY
THAT ONCOGENE DO NOT, NORMAL
TISSUES EXPRESS SOME, IT'S MANY
LOGS LOWER.
THIS IS A TRULY DEVASTATING
TUMOR, SHOWN IN GREEN ARE
PATIENTS THAT HAVE THE H3 MUTANT
VERSION.
ESSENTIALLY THEY ALL DIE.
THERE'S A MEDIAN SURVIVAL OF
NINE TO ELEVEN MONTHS.
AND IN THE MODERN DAY OF 2024,
WE HAVE NO FDA APPROVED
THERAPIES FOR THIS DISEASE.
A CHILD IS DIAGNOSED, THE
PEDIATRIC ONCOLOGIST SAYS WE'RE
GOING TO OFFER YOUR CHILD
PALLIATIVE RADIOTHERAPY, THAT'S
ALL WE HAVE TO OFFER.
SO WE DESPERATELY NEED NEW
THERAPY.
MEDIAN SURVIVAL FOR DIPG VERSION
IS NINE MONTHS.
THAT WHICH EXPRESSED HIGH
LITERALLY, MY COLLEAGUE MICHELLE
KNOCKED ON MY DOOR AND SAID
AREN'T YOU WORKING ON GD2 CAR?
MeSH DIDN'T EVEN -- MICHELLE
DIDN'T KNOW WHAT A CAR WAS, SHE
WAS A NEUROBIOLOGIST.
TRUTH BE TOLD IT'S NOT A GOOD
CAR BECAUSE WE LEARNED UNLIKE
THE CD19 CAR THAT JIM JUST
TALKED ABOUT, WHICH HAS SUCH
ASTOUNDING EFFICACY, THIS CD19
WHEN YOU MAKE IT A FLUORESCENT
CORRECTIONER YOU SEE IT
DISTRIBUTE IN THE CELL MEMBRANE
US A IMAGINE IT SHOULD BUT FOR
WHATEVER REASON THE GD2 CAR
USING THE ANTIBODY THAT IS
REALLY THE GOLD STANDARD
ANTIBODY TARGETING THIS MOLECULE
IN THE MEMBRANE LEADS TO
ACTIVATION, EVEN IN THE ABSENCE
OF ANTIGEN.
I WAS HERE AT NIH BEFORE
STANFORD IN 2016 A TALENTED
MD/PHD STUDENT WAS TRYING TO
MAKE THE CAR WORK BETTER, WE
TRIED TO USE IT FOR
NEUROBLASTOMA BUT IT SIGNALED IN
ABSENCE OF ANTIGEN, CAN YOU SEE
IT HERE, IT'S ALREADY
PHOSPHORYLATED, EVEN WITHOUT
CROSS-LINKING, DIFFERENT THAN
CD19, CELLS RAPIDLY UNDERWENT
EXHAUSTION.
THEY EXPRESSED CHECKPOINT
MOLECULES, DIDN'T WORK VERY
WELL.
WHEN WE USED THEM IN THE 28Z
CONFIGURATION.
WE USED THAT BECAUSE THAT'S THE
CONFIGURATION SURGERY BRANCH
UTILIZED.
BUT WE LEARNED IF YOU CHANGE
THAT UP, INSTEAD USE 4-1BB, A
MUTED SIGNAL, YOU COULD ACTUALLY
TURN THIS CAR FROM SOMETHING
THAT DIDN'T WORK WELL TO
SOMETHING THAT WORKED OKAY.
AND SO THIS WAS OUR OPTIMIZED
CAR THAT WE WERE WORKING WITH
WHEN I LEARNED ABOUT THE GD2
EXPRESSION AND DIFFUSE MIDLINE
GLIOMA.
USING THIS OPTIMIZED CAR, IN
NEONATAL MICE FOR WHOM THEY
RECEIVED IMPLANT INTO THE PONS
WE CURED WITH ONE IV INJECTION
OF A SMALL NUMBER OF CAR-T
CELLS, SIMILAR TO LEUKEMIA.
IT WAS SORT OF WHAT WE WERE USED
TO SEEING BUT DR. MANJAY HAD
NEVER SEEN THIS WORKED IN THIS
SPACE FOR A DECADE TRYING TO USE
SMALL MOLECULES TO CURE THIS
DISEASE.
THESE RESPONSES WERE LONG
LASTING.
THERE WAS A NOTE OF CAUTION
BECAUSE SMALL FRACTION OF MICE
DIED EARLY.
THIS WAS NOT DUE TO ON-TARGET
OFF-TUMOR TOXICITY BUT RATHER
HYDROCEPHALUS.
TUMORS IN THE PONS CELLS,
LEADING TO HYDROCEPHALUS AND
DIDN'T HAVE SUPPORTIVE CARE TO
OFFER.
WE LEARNED WE COULD FURTHER
ENHANCE THE FUNCTION OF THESE
TONICALLY SIGNALING GD2 CAR-T
CELLS IF WE MANUFACTURED THEM IN
THE PRESENCE OF A SMALL MOLECULE
INHIBITOR THAT BASICALLY
INHIBITED CAR SIGNAL, THIS BEING
DASATINIB.
SO YOU CAN SEE HERE WHEN WE
MANUFACTURED CELLS IN DASATINIB
WE WERE ABLE TO GET MORE POTENT
CELLS THAN WHEN WE ONLY USED
VEHICLE.
AND THEN FINALLY BEFORE WE TOOK
THIS TO THE CLINIC BECAUSE WE
WERE GOING TO BE TREATING A
TUMOR IN THE MIDDLE OF THE
BRAINSTEM WE KNEW THERE WERE
GOING TO BE RISKS, INCORPORATED
WHAT WE THOUGHT WAS BEST IN
CLASS SUICIDE DOMAIN, INDUCED
CASPASE 9 DEVELOPED AT BAYLOR
AND COULD BE ACTIVATED WITH
OTHERWISE INERT SMALL MOLECULE
AP 1903.
THIS WAS ABOUT 2018.
WE PUBLISHED THOSE RESULTS.
AT THE SAME TIME WHILE GETTING
CLINICAL TRIAL READY WE STARTED
TO WANT TO REALLY START TO THINK
ABOUT DIRECT DELIVERY OF THE CAR
T CELLS, BASED ON WORK COMING
OUT OF THE CITY OF HOPE WHERE
THEY WERE USING BASICALLY
INTRACEREBRAL ADMINISTRATION
INTO A CATHETER INTO THE LATERAL
VENTRICLES.
INDEED WE REPLICATED RESULTS,
QUITE REMARKABLE, IF YOU TREATED
LOCAL REGIONALLY YOU ONLY NEEDED
ABOUT A TENTH OF THE DOSE TO
CURE THE MICE AS YOU NEEDED IF
YOU TREATED INTRAVENOUSLY.
NOT ONLY DID YOU GET A MORE
POTENT ANTITUMOR EFFECT, BUT YOU
DID SO WITH LESS SYSTEMIC
INFLAMMATION, SO LOOKING AT YOUR
SYSTEMIC CYTOKINE LEVELS AGAIN
IN MICE NOW, IF YOU GIVE ICV
ADMINISTRATION YOU DIDN'T GET
THE SERUM CYTOKINES BUT GOT
SIMILAR LEVELS OF CSF CYTOKINES.
PEOPLE SOMETIMES ASK IF YOU PUT
CELLS INTO THE CSF DO THEY DIVE
INTO THE BRAIN?
THE ANSWER IS YES.
WE'VE SEEN THIS NOW BOTH IN MICE
HERE, CAN YOU SEE BY DAY 2 THE
CELLS GOING DEEPLY INTO THE
BRAIN, GREEN CELLS THERE BEING
CAR T, RED BEING TUMOR.
WE'VE ALSO NOW SEEN THIS IN
PRACTICE IN THE CLINIC.
AND SO THIS WAS WHERE WE STOOD.
WE KNEW INTRAVENOUS DELIVERY
DOING IT A SIMILAR WAY TO HOW WE
DID IT FOR LEUKEMIA OR LYMPHOMA
HAD ACTIVITY IN THE MICE BUT
ALSO HAD EARLY RESULTS
SUGGESTING DELIVERY INTO THE CNS
COULD BE USEFUL.
WE LAUNCHED A PHASE 1 CLINICAL
TRIAL, IV INFUSION CHEMOTHERAPY,
THREE DOSE ESCALATION, AFTER THE
PERIOD WAS OVER 28 DAYS WE WOULD
THEN ALLOW THE PATIENTS TO
POTENTIALLY RECEIVE A SUBSEQUENT
ICV INFUSION IN THE PATIENT
BENEFITED AND NOT HAD
DOSE-LIMITING TOXICITY.
I WANT TO CALL OUT STEVE FELDMAN
AT STANFORD, BECAUSE WE HAD
STEVE WE WERE ABLE TO GET THIS
DONE QUICKLY USING THE PRODIGY
PLATFORM, CELLS ARE READY, SEVEN
DAYS OF MANUFACTURING, MEDIAN
TIME FROM ENROLLMENT TO INFUSION
WAS 15 DAYS.
THIS WAS THE SECOND PATIENT
ENROLLED, THE FIRST WE DIDN'T
BENEFIT.
BUT THIS SECOND PATIENT CAME TO
US, MIDDLE OF THE PANDEMIC, JUNE
2020, A MEDICAL STUDENT DROVE
FROM FLORIDA TO CALIFORNIA, WAS
ABLE TO WALK WHEN THE DRIVE
STARTED BUT TUMOR IN THE SPINAL
CORD, NOT ABLE TO WALK BY THE
TIME SHE ARRIVED.
SHE HAD RAPIDLY PROGRESSING
TUMOR.
BECAUSE OF RAPID PROGRESSION
RETREATED HER ON COMPASSIONATE
EXEMPTION, WOULDN'T BE ABLE TO
DISTINGUISH TOXICITY FROM
PROGRESSION.
NONETHELESS SHE RECEIVED THE
THERAPY AS DIRECTED IN THE
PROTOCOL, WE SAW DRAMATIC
SHRINKAGE OF THE LARGE SPINAL
TUMOR, ASSOCIATED WITH CLINICAL
IMPROVEMENT.
SHE WAS COMPLETELY PARALYZED
WHEN SHE WENT ON WITH THIS
REDUCTION WAS ABLE TO WALK WITH
A LOT OF ASSISTANCE BUT ALSO
WENT FROM HAVING COMPLETE LOSS
OF BOWEL BLADDER FUNCTION TO
RETRIEVING BLADDER FUNCTION, THE
FIRST SIGNAL THAT MAYBE WE HAVE
SOMETHING HERE.
HERE'S THE THIRD PATIENT, A
YOUNG ADULT WITH SOMEWHAT
UNUSUAL, PONTINE TUMOR HAD A
CYST IN THE MIDDLE, COMPRESSED
FOURTH VENTRICLE, CAUSING
MASSIVE PRESSURE, GAVE
INTRAVENOUS INFUSION.
SYMPTOMS WERE MAINLY TROUBLE
OPENING HIS MOUTH, HE COULD OPEN
MIDWAY BUT NOT NORMALLY.
AND SEVEN DAYS AFTER INFUSION HE
HAD GONE FROM BEING ABLE TO OPEN
MIDWAY TO BASICALLY NOT BEING
ABLE TO OPEN AT ALL.
THIS WAS AT THE TIME OF A PEAK
CYTOKINE RELEASE SYNDROME.
SYMPTOMS WERE GETTING WORSE.
THIS WAS EXACTLY WHAT WE
ANTICIPATED BASED ON WHAT WE
LEARNED IN THE MOUSE, THE TUMOR
WAS SWELLING, SO HE USED A STRAW
TO DRINK, BY DAY 14 WAS NOW
STARTING TO BE ABLE TO USE HIS
MOUTH AGAIN, WE SAW SOME OPENING
UP OF THIS FOURTH VENTRICLE, NOW
STARTS TO HAVE A NORMAL SHAPE,
NUMBNESS ON THE LEFT SIDE OF HIS
FACE IMPROVING, BY DAY 21
SYMPTOMS ON FACE HAD RESOLVED.
HE WAS ABLE TO EAT A LARGE
CHEESEBURGER AND BOY, WAS HE
HAPPY.
SO IT REALLY WAS QUITE
REMARKABLE TO SEE THIS TOXICITY
SYNDROME WHICH WE'VE NOW CALLED
TUMOR INFLAMMATION ASSOCIATED
TOXICITY, REMINED ME BACK IN THE
DAYS WHEN CYTOKINE RELEASE
SYNDROME WAS DESCRIBED WE NEEDED
TO UNDERSTAND TOXICITY, TO GRADE
IT, AND TO BE ABLE TO REPORT THE
TOXICITY AND ALSO MANAGE IT IN A
GRADE-DIRECTED WAY.
WE WORKED WITH INVESTIGATORS
ACROSS THE COUNTRY TREATING
BRAIN TUMORS WITH VARIOUS TYPES
OF CELL THERAPIES INCLUDING CD19
CAR FOR CD19 POSITIVE TUMORS IN
THE BRAIN, COINED THIS TOXICITY
SYNDROME DISTINCT FROM CRS,
CAUSING LOCAL NEUROLOGIC
DYSFUNCTION OR MORE SEVERE CASES
HYDROCEPHALUS WHICH WOULD
REQUIRE URGENT INTERVENTION.
WE LAID OUT THE MANAGEMENT.
COULD BE OBSERVATION AND
SUPPORTIVE CARE ONLY TO RAPID
INTERVENTION TO LOWER
INTRACRANIAL PRESSURE.
BY UNDERSTANDING THE
PATHOPHYSIOLOGY, AND GRADING IT,
WE WERE ABLE TO BASICALLY KEEP
OUR TRIAL OPEN, NOT BE SHUT DOWN
FOR TOXICITY WE DIDN'T
UNDERSTAND, AND IMPORTANTLY WERE
ABLE TO KEEP ALL THESE PATIENTS
SAFE.
I THINK THIS IS PART OF THE WAY
WE DEVELOPED NEW THERAPIES, BY
NOT SHYING AWAY FROM TOXICITY
BUT LEANING IN AND REALLY
UNDERSTANDING WHAT IS HAPPENING
AND CAN WE MANAGE OUR WAY
THROUGH IT.
SO AT THE END OF ARM A, ARM A IS
THIS IV INFUSION FOLLOWED BY ICV
GIVEN AS EARLY AS EVERY MONTH,
WE SAW THAT THE FIRST INFUSION
WAS THE MOST TOXIC ONE, IT WAS
THE IV ONE, SAW SIGNIFICANT
CYTOKINE RELEASE SYNDROME,
DOSE-LIMITING TOXICITY DUE TO
CRS AND PATIENTS DON'T TOLERATE
THAT WELL.
WE SAW VERY LITTLE ICAN
SURPRISING.
WE SAW TUMOR ASSOCIATED
TOXICITY, NEUROTOXICITY, MORE
COMMON EARLY ON, AND GRADUALLY
DIMINISHED LIKELY BECAUSE THERE
WAS MORE SPACE AS THE TUMOR WAS
RESPONDING.
WE NEVER SAW A DLT AFTER THE ICV
INFUSION, REMARKABLY SAFE.
THIS IS A WATERFALL PLOT, A
TUMOR FOR WHICH WE'VE HAD NOT
THERAPY, NOW FOUR PATIENTS WITH
50% VOLUMETRIC REDUCTION, ALL
PROGRESSING AT THE TIME OF
ENROLLMENT.
THIS PATIENT HAD A COMPLETE AND
DURABLE RESPONSE, I'LL TALK
ABOUT HIM.
SOME HAVE WONDERED IS HE AN
OUTLIER.
HE'S NOT AN OUTLIER.
IN FACT IF YOU LOOK AT THE
OVERALL RESPONSE FROM THE
VOLUMETRIC CHANGE, WE HAVE
GAUSSIAN DISTRIBUTION, A BROADLY
ACTIVE THERAPEUTIC THAT IS, YOU
KNOW, REALLY SHOWING
REPRODUCIBLE AND IN SOME CASES
DURABLE ACTIVITY.
THE MEDIAN OVERALL SURVIVAL,
REMEMBER I TOLD YOU 9 TO 11
MONTHS FOR THESE PATIENTS, IT
WAS 17 MONTHS FOR THE DIPG
PATIENTS, IF YOU INCLUDE SPINAL
DMGs, 20.6 MONTHS.
SO, MOST OF THESE PATIENTS WENT
ON TO DIE EXCEPT FOR THE PATIENT
10, I'LL TELL YOU ABOUT, BUT FOR
THIS DISEASE SEEING SIGNIFICANT
CLINICALLY RELEVANT ACTIVITY AND
CHANGES IN SURVIVAL WAS VERY
ENCOURAGING.
HERE'S PATIENT NUMBER 3, WHO HAD
THAT ORIGINAL NICE RESPONSE,
LASTED ABOUT THREE MONTHS.
HE THEN CAME BACK WITH
DIFFICULTY SMILING.
AGAIN HIS SYMPTOMS ON THE LEFT
SIDE OF HIS FACE WERE RECURRENT.
IN THIS TIME WE GAVE HIM ICV
INFUSION, YOU CAN SEE IT MADE
HIS FACIAL MUSCLES ABLE TO SMILE
AGAIN, HERE HE IS WHEN HE CAME
BACK WITH REALLY MORE
RIGHT-SIDED STIFFNESS, AND
SPASTICITY, DIFFICULTY WITH
BALANCE, WE GAVE HIM THE ICV
INFUSION, HERE IS HE IN CLINIC,
A FEW DAYS LATER ABLE TO WALK
REALLY QUITE WELL, TOOK A DAY
HIKE WITH HIS SISTER.
EVENTUALLY ABOUT A YEAR AND A
HALF IN HE DIED OF A HEMORRHAGE
IN HIS TUMOR WHICH IS SEEN WITH
THESE TUMORS.
BUT WE CLEARLY GAVE HIM ABOUT A
YEAR AND A HALF OF GOOD TIME.
HERE'S PATIENT NUMBER 10, A
17-YEAR-OLD WHO CAME TO US WHEN
HE WAS A JUNIOR IN HIGH SCHOOL
WITH I HAD NASTY-LOOKING LARGE
PONTINE TUMOR, FOURTH VENTRICLE,
COMPLETE RESPONSE OVER ABOUT
FIVE MONTHS.
NOW SUSTAINED FOR MORE THAN 3
1/2 YEARS.
WHEN HE CAME, HE WAS ABLE TO
WALK BUT IF HAD HE TO GO TOO FAR
WAS IN A WHEELCHAIR, COULDN'T
ATTEND HIGH SCHOOL, HE IS NOW
STARTING ACTUALLY A SENIOR IN
COLLEGE NOW, SPENT THE SUMMER
TRAVELING EUROPE WITH HIS
FAMILY.
HERE HE IS A MONTH AFTER HIS
FIRST INFUSION, YOU CAN SEE HE'S
WITH HIS FATHER, WHO IS HELPING
HIM WALK BECAUSE HE CAN WALK,
BUT NOT VERY WELL.
AND HERE HE IS EIGHT MONTHS
AFTER FIRST INFUSION.
SO DRAMATIC REALIZATION FROM THE
TRIAL MUCH OF THE NEUROLOGIC
FUNCTION LOST IS POTENTIALLY
RECOVERABLE, AND HE'S NOW ABLE
TO RUN.
HERE IS LOOKING AT CLINICAL
BENEFIT IN BLUE, CAN YOU SEE
MANY OF THESE PATIENTS HAD
SIGNIFICANT CLINICAL BENEFIT,
AND SOME PATIENTS WHO DIDN'T
EVEN HAVE SHRINKAGE OF TUMOR ON
THE SCANS IT'S VERY HARD TO KNOW
WHAT'S TUMOR AND WHAT'S THE PONS
BECAUSE IT'S INFILTRATIVE BY
NATURE BUT THIS GIRL WAS 3 WHEN
SHE WENT ON, HAVING TROUBLE
COLORING BECAUSE OF THE
DYSFUNCTION, AND EVEN FOUR WEEKS
IN WAS ABLE TO REALLY HAVE MUCH
BETTER FINE MOTOR COORDINATION
AND SHE ENDED UP LIVING FOR OVER
TWO YEARS ON TRIAL WITH
GENERALLY GOOD QUALITY OF LIFE.
WHY DID SOME RESPOND AND NOT
OTHERS IN THE MILLION DOLLAR
QUESTION.
THIS IS THE DURABLE COMPLETE
RESPONSE, TUMOR VOLUME GOES TO
ZERO.
HIS CELL-FREE DNA AND CNS GOES
TO ZERO, PERSISTING IN
BLOODSTREAM OVER A YEAR OUT.
WE DON'T KNOW IF THESE ARE THE
CELLS HE GOT IV OR CELLS WE GAVE
ICV, WE KNOW ADMINISTRATION ICV
CAN LEAD TO TRAFFICKING IN THE
BLOOD BUT HE HAD THE MOST
DURABLE PERSISTENCE OF CAR-T
CELLS.
WE ALSO KNOW THAT EVERY TIME WE
GAVE HIM AN INTRACRANIAL
INFUSION WE COULD HAVE
INFLAMMATORY SIGNATURE IN HIS
CSF, LARGELY DRIVEN BY
INTERFERON GAMMA AND INDUCED
CYTOKINES.
HERE IS PATIENT NUMBER 10 WITH
THAT DURABLE PERSISTENCE OF CAR
Ts, ANOTHER YOUNG WOMAN SHE
HAD A 99% RESPONSE, TUMOR
COMPLETELY DISAPPEARED EXCEPT
FOR ONE TEENY SPOT, AND THEN
WOULDN'T YOU KNOW IT, EVENTUALLY
STARTED TO REGROW.
AND WE DON'T KNOW WHY BUT WE DO
KNOW THE TIME OF REGROWTH
COINCIDED WITH LOSS OF HER CAR T
CELL PERSISTENCE.
WE HAVE SEEN SOME -- BEEN ABLE
TO LOOK AT SOME TUMORS AT THE
TIME OF PROGRESSION AND SO FAR
WE'VE NOT SEEN LOSS OF TARGET.
WE KNOW RESISTANCE CAN HAPPEN
WITHOUT TARGET LOSS.
THAT DOESN'T MEAN NO ONE HAS
LOST TARGET.
WHEN YOU LOOK AT WHAT'S GOING ON
IN THE CSF IT'S REALLY
REMARKABLE WE'VE BEEN LEVERAGING
SINGLE CELL RNAseq IN THE
RESERVOIR, ACCESSING THE CELLS,
MANY OF THEM ARE T CELLS BUT
MANY OF THESE ARE MYELOID CELLS
IN THE CSF.
MYELOID CELLS COME IN MANY
SHAPES AN SIZES.
SOME ARE DERIVED FROM THE
MONOCYTES IN THE BLOOD.
SOME DERIVED FROM MICROGLIA,
SOME IMMUNE ACTIVATING, SOME
IMMUNE SUPPRESSIVE.
A TEMPORAL COURSE SEEMS TO WORK
WITH EACH PATIENT FOLLOWING A
SIMILAR TEMPORAL COURSE WITH
INFLUX OF MYELOID CELLS, COULD
BE WHY WE'RE SEEING RESISTANCE,
WE DON'T KNOW.
WE HAVE A CLEAR SIG AND -- SIG.
NOT A HOME RUN BUT WE'RE ON
BASE.
WE CONTINUE TO ACCRUE TO PHASE 1
TRIAL.
I SHARED RESULTS FROM ARM A.
WE ENROLLED PATIENTS ON ARM B
WHERE WE USE NO LYMPHODEPLETION,
ICV ADMINISTRATION ONLY, HIGH
RATES OF RESPONSE BUT RESPONSE
WAS NOT DURABLE.
WE'RE WORRIED MAYBE THESE
PATIENTS WERE REJECTING CAR-T
CELLS BUT DON'T HAVE PROOF AS
YET.
WE'RE NOW TREATING ON ARM C, WE
GIVE ICV DELIVERY AND PERIODIC
DEPLETION.
WE'VE SEEN IMPRESSIVE RESPONSES
WITH DURABILITY, MAYBE THAT'S
GOING TO BE THE FINAL DOSE ROUTE
AND SCHEDULE FOR FORMAL EFFICACY
TESTING.
I DIDN'T TALK ABOUT SAFETY
MANAGEMENT BUT IT'S EVOLVES
SUBSTANTIALLY, WE'RE CONFIDENT
IN OUR APPROACH AT THE SINGLE
INSTITUTION LEVEL BUT IMPORTANT
NEXT STEPS TO EXPORT TO OTHER
PEDIATRIC HOSPITALS TO BE SURE
OTHERS CAN DELIVER IT SAFELY AND
WE'RE NOW EASING OUR ELIGIBILITY
CRITERIA TO SOME MORE HIGH RISK
CEREBELLAR AND THYLAMIC
PATIENTS.
WE'RE PROPOSING MULTI-CENTER
POTENTIALLY PIVOTALL PHASE 2
PRIAL, -- TRIAL, ENPOINTS COULD
BE RESPONSE RATE, OR SURVIVAL.
A DRUG WAS APPROVED FOR
NEUROFIBROMATOSIS PATIENT ON
PATIENT-REPORTED OUTCOMES SO
WE'RE INCORPORATING PATIENT
OUTCOMES INTO THE TRIAL NOW BUT
THE BIGGEST PROBLEM THAT KEEPS
ME UP AT NIGHT WE HAVE NO WAY TO
DEVELOP THIS THROUGH THE
COMMERCIAL MODEL.
BIOPHARMA WILL NOT DEVELOP CELL
AND GENE THERAPIES FOR
PEDIATRICS.
SO WE NEED NEW BUSINESS MODELS
TO BE ABLE TO DELIVER THIS TO
PATIENTS, IN A WAY THAT ENSURES
ACCESS TO EVERY PATIENT WHO
NEEDS IT.
AND SO THAT'S WHAT I FIND MYSELF
WORKING ON TODAY.
A NEW BUSINESS MODEL.
SCIENTIFIC WORK IS ONGOING, WE
NEED NEXT GENERATION APPROACHES,
WE'RE DOING THAT IN PARALLEL.
WHY THIS DISEASE?
WE HAVE AN ONCOGENE-DRIVEN
DRIVER, THIS MUTATION DRIVES GD2
EXPRESSION.
WE HAVE AN OPTIMIZED CAR.
WE HAVE LOCAL REGIONAL
ADMINISTRATION THAT COULD
POTENTIALLY OVERCOME TRAFFICKING
PROBLEMS.
BUT IF WE HAD HAD ONLY THE
ABILITY TO GIVE ONE DOSE I THINK
WE WOULD HAVE SEEN HIGH RESPONSE
RATES AND HIGH RECURRENCE RATES.
THE FACT THAT WE'RE ABLE TO GIVE
REPEATED DOSING IN THESE
DISEASES THAT ARE VERY STEM-LIKE
I THINK ARE VERY IMPORTANT.
I DON'T KNOW IF SOLID TUMORS
WILL BE ABLE TO BE CURED WITH A
ONE AND DONE APPROACH LIKE
LIQUID TUMORS.
IT TAKES AN ARMY. .
I'VE HIGHLIGHTED THE WORK.
THE ENTIRE TEAM AT STANFORD AND
PATIENTS AND FAMILIES, AND HERE
ARE OUR FUNDERS.
THANK YOU VERY MUCH.
[APPLAUSE]
>> IF WE DON'T HAVE QUESTIONS I
WOULD LIKE TO THANK AGAIN
EVERYONE FOR PARTICIPATING IN
DAY 1.
WE WILL ADJOURN.
REMINDER FOR THOSE THAT ARE
SURGERY BRANCH ALUMNI, WE'RE
GOING TO GATHER ON THE FAES
TERRACE WHERE POSTER SESSION WAS
AT AN ATTEMPT AT GROUP PHOTO.