>>I'M DELIGHTED TODAY AND
HONORED, REALLY, TO INTRODUCE
DR. TONI RIBAS.
HE IS A PROFESSOR OF MEDICINE
WITH TENURE, PROFESSOR OF
SURGERY IN MOLECULAR AND MEDICAL
PHARMACOLOGY AT UCLA AND THE
DIRECTOR OF THE TUMOR IMMUNOLOGY
PROGRAM AT THE JONSSON
COMPREHENSIVE CANCER CENTER, AND
DIRECTOR OF THE PARKER INSTITUTE
FOR CANCER IMMUNOTHERAPY AT
UCLA.
TONY DID HIS MD PH.D. AT THE
UNIVERSITY OF BARCELONA AND
STAYED ON AS A CLINICAL
INSTRUCTOR BEFORE GOING TO UCLA
IN 1996 AND DOING HIS POSTDOC
THERE.
SINCE THAT TIME, HE STAYED AT
UCLA IN MULTIPLE DIFFERENT ROLES
AND HE REALLY IS A TRUE
PHYSICIAN SCIENTIST.
HIS RESEARCH IS FOCUSED ON
LABORATORY AND CLINICAL STUDIES
TO ADDRESS PRIMARY AND ACQUIRED
RESISTANCE FOR IMMUNOTHERAPY AND
TARGETED THERAPIES AND ALSO THE
DEVELOPMENT OF GENE ENGINEERED
ADOPTIVE T-CELL TRANSFER
THERAPIES FOR OTHER CANCERS.
HE'S BEEN INSTRUMENTAL IN THE
CLINICAL DEVELOPMENT OF SEVERAL
DIFFERENT AGENTS THAT HAVE LED
TO FDA APPROVAL, INCLUDING
PEMBROLIZUMAB, THE FIRST PD-1
INHIBITOR APPROVED BY THE FDA,
TWO COMBINATIONS OF BRAF AND MEK
INHIBITORS AND WAS ALSO INVOLVED
IN THE INITIAL CD-19 CAR T FOR
LYMPHOTHAT AXI CELL.
HE'S A PAST PRESIDENT OF THE
AACR AND ELECTED FELLOW IN THE
AACR ACADEMY, THE ROYAL ACADEMY
OF MEDICINE OF CATALONIA, THE
AMERICAN SOCIETY OF CLINICAL
INVESTIGATION AND THE NATIONAL
ACADEMY OF MEDICINE.
YOU KNOW, MY FIRST INTRODUCTION
TO TONY REALLY WAS WHEN I HAD
THE PRIVILEGE OF BEING
ASSOCIATED WITH HIM AS HE WAS
THE CO-LEADER OF THE STAND UP TO
CANCER IMMUNOTHERAPY DREAM TEAM,
ALONG WITH DR. JIM ALISON.
AND YOU KNOW, I WAS REALLY
IMPRESSED BY HIS LEADERSHIP AND
HIS SCIENTIFIC INSIGHTS THERE.
I ALSO HAD THE DISTINCT -- AND I
WOULD GO AND VISIT WITH THE
REVIEW TEAM EVERY SIX MONTHS FOR
ABOUT FIVE YEARS, AND THEY MADE
SOME AMAZING PROGRESS, HAD SOME
AMAZING SCIENCE THAT CAME OUT OF
THAT TEAM.
IN ADDITION, I HAD THE DISTINCT
PLEASURE OF WORKING WITH HIM ON
THE AACR NCI EURTC TRIPLE
MEETING FOR TWO YEARS AND
REALLY, I JUST WANT TO SAY THAT
HE'S AN INCREDIBLY BRIGHT AND
THOUGHT-PROVOKING LEADER, HE'S A
DYNAMIC SPEAKER, AND HE'S A
SINCERE PERSONAL FRIEND.
SO PLEASE JOIN ME IN WELCOMING
DR. TONY RIBAS.
[APPLAUSE]
>>THIS IS WHAT HAPPEN WHEN IS A
FRIEND INTRODUCES YOU AND MAKES
YOU LOOK GOOD.
SO I WISH MY MOM WAS HERE AND
SHE COULD HEAR THIS.
[LAUGHTER]
I'LL START WITH MY DISCLOSURES.
WHAT I'M TRYING TO TALK ABOUT IS
PERSONALIZED CELLULAR THERAPIES
FOR CANCER.
I'VE BEEN INVOLVED WITH SEVERAL
COMPANIES.
THERE'S ONE THAT HAS A
PARTICULAR RELEVANCE, WHICH IS
THIS 1PACPHARMA, BECAUSE ALL OF
THE WORK I'M GOING TO BE TALKING
ABOUT IS IN COLLABORATION WITH
THEM.
SO PLEASE TAKE THAT INTO ACCOUNT
ON WHAT I'M TALKING ABOUT.
AND I'M GOING TO BE TALKING
ABOUT ANALYSIS OF ANTIGEN
SPECIFIC -- BY TAKING SAMPLES
FROM PATIENTS, ANALYZING THIS
FIELD OF RARE CELLS THAT HAVE
ANTIGEN SPECIFICITY FOR THE
CANCER.
AND THEN THE ABILITY TO
RECOGNIZE THOSE CELLS ALLOWS US
TO THINK ABOUT THE THERAPEUT%C
WHERE WE ENGINEER CELLS TO
EXPRESS THE RECEPTORS AND GIVE
THEM TO PATIENTS WITH CANCER.
I KNOW IT TAKES A LOT OF COURAGE
TO GO TO THIS BUILDING AND TALK
ABOUT TARGETING NEOANTIGENS, AND
IT PROBABLY WHY JAMES HAS
INVITED ME ON A DAY THAT --
ROSENBERG, IT'S ONE OF THE FEW
DAYS HE'S NOT IN TOWN.
STEVE TOLD ME I'M ON VACATION IN
LOS ANGELES AND YOU'RE COMING
HERE, SO IT GIVES ME A LITTLE
BIT MORE CONFIDENCE OF WHAT I'M
TALKING ABOUT, BECAUSE WE'VE
STUDIED ALL OF THESE BECAUSE OF
THE PIONEERING WORK THAT HAS
BEEN DONE IN THIS BUILDING,
ABOUT HOW TILLS RECOGNIZE
CANCER -- AT THE CORE OF HOW THE
IMMUNE SYSTEM CAN DIFFERENTIATE
CANCER CELLS FROM NORMAL CELLS.
OUR THOUGHT WAS, WELL, IF THAT'S
THE CASE AND WE KNOW THERE'S THE
ALPHA AND BETA T-CELL RECEPTOR,
CAN WE CLONE THOSE T-CELL
RECEPTORS AND ENGINEER OTHER
CELLS IN A WAY OF MAKING
SYNTHETIC TILL THERAPIES.
SO TO DO THAT, WE HAD TO COME UP
WITH A SERIES OF NEW
TECHNOLOGIES TO BE ABLE TO STUDY
THE LANDSCAPE OF RESPONSES TO
NEW ANTIGENS, CLONE THOSE T-CELL
RECEPTORS AND ENGINEER THEM
BACK.
ONE OF THE NEW TECHNOLOGIES THAT
NEEDED TO BE GENERATED IS TO BE
ABLE TO ISOLATE PERSONALIZED
T-CELL RECEPTORS BY DIRECT
MISSION OF THE NEOANTIGENS, BUT
WE WANT TO NOT DO IT ONLY ON
HLA2.1, WE WANT TO BE ABLE TO
COVER THE MAJORITY OF THE
POPULATION, AND TO DO THAT, WE
WANTED TO HAVE A PLATFORM THAT
WOULD ALLOW US TO DO AS MANY
HLAs CLASS Is AS WE COULD,
ENDED UP 64 C CLASS Is, AND BE
ABLE TO TAKE THOSE T-CELL
RECEPTORS AND ENGINEER THEM BACK
INTO THE PATIENT'S OWN T-CELLS,
AND IF WE TRIED TO DO THAT WITH
RETROVIRUS AND LENTIVIRUSES, WE
COULD TREAT ONE PATIENT, I DON'T
KNOW HOW MANY -- IN HOW MANY
YEARS, AND THEN DO ANOTHER ONE,
WE WANTED TO DO THIS IN AN
EFFICIENT WAY SO WE HAD TO
CREATE THOSE TECHNOLOGIES.
SO I'LL WALK YOU THROUGH THE
FIRST GENERATION OF THE
TECHNOLOGIES AND THEN I'LL WALK
YOU THROUGH WHAT EVENTUALLY WAS
DONE WHEN THE TECHNOLOGIES WERE
INDUSTRIALIZED AND PUT INTO
CLINICAL -- THE FIRST WAS A
POSTDOC IN JIM KEITH'S LAB,
POSTDOC IN DAVID BALTIMORE'S LAB
AND A GRADUATE STUDENT IN MY LAB
WHO WORKED ON THIS SOLUBLE
COMPLEXES OF -- PEPTIDE FROM DON
SCHUMACHER'S APPROACH AND LOAD
IT WITH A MUTATIONAL PEPTIDE,
PUT IT INTO A NANOPARTICLE AND
SORTED THEM OUT.
THAT'S NOT WHAT WE DID IN THE
CLINIC.
THIS WAS THE FIRST ITERATION, IT
NEEDED TO BE MORE SOLID THAN
WHAT WE DESCRIBED IN THIS PAPER.
AND THEN IF WE DO THAT, WE
ISOLATE T-CELLS WITH THAT --
THAT RECOGNIZE TUMOR ANTIGENS BY
THIS SORTING WITH THE HLA
PEPTIDE COMPLEXES, THEN WE NEED
TO GET THOSE T-CELL RECEPTORS
BACK.
AND TO DO THAT, WE HAVE TO ALSO
GENERATE THE NEW TECHNOLOGY, AND
THAT WAS ENABLED BY THE CRISPR
EDITING WHERE YOU CAN ONE STEP,
YOU CAN -- CAS9 ON A DNA PLASMIC
WITH HOMOLOGY ENDS THAT WOULD
BIND TO WHERE YOU CUT IT AND YOU
COULD EFFICIENTLY KNOCK OUT THE
GENE AND KNOCK IN ANOTHER GENE.
AND THIS WOULD BE THE T-CELL
RECEPTOR.
THE FIRST ITERATION OF THIS WAS
DONE BY A GRADUATE STUDENT IN
ALEX MARSON'S LAB AND A POSTDOC
IN MY LAB, AND YOU'LL SEE MORE
WORK FROM CHRIS THAT COMES FROM
THIS ABILITY TO --
SPECIFICALLY -- IN AN EFFICIENT
MANNER.
SO THEN WE'VE DONE ALL OF THIS,
THEN I'M GOING TO SHOW YOU IF
THE TECHNOLOGIES WORK, WHAT DO
THESE T-CELL RECEPTORS DO, CAN
WE GET THESE T-CELLS THAT ARE
GENE-ENGINEERED WITH A NEO --
THAT WE SORTED FROM PATIENTS,
CAN WE GET THEM TO SHOW THEY
RECOGNIZE AND KILL TUMOR CELLS.
SO I'M GOING TO GO THROUGH DATA
PRESENTED BY CRIS THAT PAST FALL
THAT'S NOW BEEN WRITTEN UP AND
IT WILL COME OUT IN PUBLICATION.
THIS IS THE ABILITY TO USE THESE
TECHNOLOGIES TO STUDY AN IMMUNE
RESPONSE IN THA HUMANS.
WE'LL TALK AFTERWARDS HOW TO USE
THE T-CELL RECEPTORS AS
THERAPEUTICS.
WHAT WE WANTED TO DO WAS HAVE A
LANDSCAPE ANALYSIS OF
NEOANTIGEN-SPECIFIC T-CELL
RESPONSES INDUCED BY IMMUNE
CHECKPOINT BLOCKADE.
AND TO DO THIS, WE TOOK 11
PATIENTS, ALL OF THEM FROM MY
CLINIC AT UCLA, THAT HAD
RECEIVED ANTI-PD-1-BASED THERAPY
IN SOME FORM, DIFFERENT
ANTI-PD-1s, MOST OF THEM
SINGLE AGENTS, SOME OF THEM IN
COMBINATION.
OF THESE 11 PATIENTS, THE FIRST
SEVEN PATIENTS HAD OBJECTIVE
RESPONSES TO THERAPY.
AND THE LAST FOUR HAD NO
RESPONSES TO THERAPY.
SO WE WANTED TO COMPARE THE
T-CELLS THAT WE ISOLATED FROM
THESE PATIENTS FROM T-CELLS THAT
WE ISOLATED FROM THESE PATIENTS.
AND IN SOME OF THEM, IN FIVE OF
THESE 11 PATIENTS, WE HAD
STUDIED A PATIENT
MELANOMA-SPECIFIC CELL LINE,
TESTING IN VITRO AGAINST THE
PATIENT'S OWN CELL LAN
IRRESPECTIVE OF WHAT HLA WAS
RESTRICTED BECAUSE THEY WERE ALL
ENDOGENOUS HLAs.
SO NOW THE TECHNOLOGY AS IT
EVOLVED WHEN PACPHARMA PUT OVER
100 PEOPLE TO WORK ON IT, WAS
THAT WE WOULD TAKE A TUMOR AND
CELL LINE INITIALLY, ANY TUMOR
BIOPSY, AND DO DNA TO LOOK FOR
MUTATIONS, RNA TO LOOK FOR
EXPRESS GENE, AND THEN I COMPARE
IT WITH A PERIPHERAL BLOCK MONO
NUCLEAR CELLS, SO YOU COULD HAVE
A PREDICTED LIST OF MUTATIONS
THAT COULD -- THAT WOULD BE
PRESENTED BY DIFFERENT HLAs
AND THE SEQUENCING WOULD GIVE US
THE SIX HLAs OF THE PATIENT,
AND THEN WE'D HAVE THIS ABILITY
TO MAKE PROTEINS THAT HAVE --
THAT ARE A SINGLE CHAIN TRIMER
OF HLA BETA TO MICRO GLOBULIN ON
THE MUTATED PEPTIDE ALL IN A
SEQUENCE OF NUCLEOTIDES THAT
THEN WOULD MAKE THE PROTEIN IN
293 CELLS, AND THOSE PROTEINS
WOULD SELF-ASSEMBLE IF THE
PEPTIDE STABILIZES, IT
STABILIZES THIS SINGLE CHAIN
TRIMER WITH BIOTIN LAITION AND
THEN YOU HAVE THE AFFINITY, DNA
BARCODING TO HAVE HUNDREDS OF
THESE PER PATIENT, FLUORESCENTLY
LABELED AND COCULTURED WITH
T-CELLS THAT WE OBTAIN FROM THE
PATIENT'S BLOOD OF THE PATIENTS'
TUMORS.
I KNOW I'M GOING FAST ON ALL OF
THIS, BUT IT A COMPLICATED
TECHNOLOGY, BUT THE WHOLE THING
IS THAT WE CAN MAKE THESE
BARCODED HLA MUTATED PEPTIDE
COMPLEXES THAT COULD BE USED
THROUGH MULTIPLE HLAs, THE SIX
HIGH PRESSUHLAs FROM EVERY PATI.
SO HERE ARE THE 11 PATIENTS, AND
THIS IS THE NUMBER OF MUTATIONS.
HERE ARE THE PATIENTS WHO HAVE
RESPONDED TO ANTIPD-1 THERAPY,
AND HERE ARE PATIENTS WHO DID
NOT RESPOND TO ANTIPD-# THERAPY.
THESE ARE ALL PATIENTS WITH
MELANOMA.
THE PATIENTS WITH CLINICAL
RESPONSE TEND TO HAVE MORE
MUTATIONS BULL I BUT IT'S NOT L.
THESE HAD 977 NON-SYNONYMOUS
MUTATIONS.
FROM THERE, WE LOOKED AT THE
MUTATIONS THAT COULD BE SCREENED
BECAUSE WE HAD THE PREDICTION
ALGORITHMS TO PRESENT WITH ONE
OF THESE 64 HLA CLASS 1s THAT
IN THE PACKPHARMA REPOSITORY,
THEN THEY MADE EIGHT OF THESE
SINGLE CHAIN TRIMERS THAT ARE
MULTI-MERRIZED OF NEOANTIGEN
HLAs.
SO FROM THIS FIRST PATIENT THAT
THAT -- MADE IT INTO 243 LIBRARY
OF HLA PEPTIDE -- BETA PEPTIDE
COMPLEXES BECAUSE FOR SOME
MUTATIONS, YOU COULD HAVE A
PRESENTATION BY DIFFERENT
HLAs.
WE CAN GO DOWN, THIS IS THE
MAXIMUM THAT WE DID IS 243.
THIS TUMOR THAT HAD MUTATION, WE
COULD ONLY MAKE 17 SOLID
COMPLEXES OF PEPTIDE HLA.
SO WE SEE HOW MANY OF THESE
COMPLEXES BOUND TO T-CELLS IN
PATIENTS' BLOOD THAT MADE THE
CUTOFF TO SAY THAT THEY'RE
ANTIGEN-SPECIFIC T-CELLS.
WE CAN SEE THAT NOW THE NUMBER
OF MUTATIONS THAT ARE TARGETED
AND RECOGNIZED BY T-CELLS ARE
VERY FEW.
SO EVEN IF YOU START WITH 3,000,
THERE WERE ONLY -- WE MADE A
LIBRARY OF 243 HLA PEPTIDE
COMPLEXES, ONLY 11 OF THEM WERE
RECOGNIZED.
LET GO DOWN HERE FROM 17 PEPTIDE
HLA COMPLEXES, FIVE WERE
RECOGNIZED.
TELLING US THAT THERE'S
SOMETHING CONSTANT HERE WHERE
THE IMMUNE SYSTEM RECOGNIZES
SOME MUTATIONS IN AN
IMMUNODOMINANT WAY, AND SEEMS TO
IGNORE MANY OTHERS, AND IT'S NOT
LINEAR LIQUOR LALY CORRELATED, R
THAT THE TUMOR HAS AND THE
NUMBER OF T-CELLS THE PATIENT
RECOGNIZES.
THESE DID NOT SEPARATE PATIENTS
WITH RESPONSE OR NO RESPONSE TO
THERAPY.
JUST THE METRIC OF THE NUMBER OF
T-CELLS RECOGNIZING THESE
COMPLEXES.
BUT IF WE LOOK AT THE NUMBER OF
P CELL RECEPTOR CHRONOTYPES THAT
RECOGNIZE THESE MUTATIONS, THEN
THERE'S A SPREAD.
THE PATIENTS WHO HAD CLINICAL
RESPONSE TO THERAPY, THEY HAD A
LOT MORE CLONAL TYPES FOR EVERY
KNEW TAITIONEVERYMUTATION THAT'.
SO SAY THERE'S THREE RECOGNITION
BUT 24 DIFFERENT T-CELL
RECEPTORS THAT RECOGNIZE THOSE
THREE HLA PEPTIDE COMPLEXES.
IN PATIENTS WITHOUT RESPONSE TO
THERAPY, THERE WAS SOME
DISCREPANCY BUT THE MAJORITY OF
THEM HAD A LOT LESS TCR CLOA
KNOW TYPES RECOGNIZING SIMILAR
NUMBERS OF MUTATIONS.
THE DATA I'M SHOWING YOU, I'M
GOING TO WALK YOU THROUGH HERE.
THIS IS A PATIENT WITH A LUNG
METASTASES RESPONDING TO
ANTI-PD-1.
THIS IS PRUF RAL BLOOD MONO --
BASELINE AT DAY 14.
HERE FROM THIS PATIENT NUMBER
TWO, WE HAD MET A LIBRARY OF 24A
COMPLEXES, I THINK IT WAS OVER
FOUR OF THE SIX, AND THEY WERE
ORGANIZED BY LOW AFFINITY TO
HIGH AFFINITY.
AND THIS IS THE RECOGNITION OF
T-CELLS, SO EACH BOX IS ONE
T-CELL.
AND X IS 10 T-CELLS SO WE DON'T
DWARF ALL OF THESE.
THERE'S ACTUALLY THREE MUTATIONS
THAT ARE RECOGNIZE DOLLARS.
IL-18 PRESENT BY HLA2.4 OR HLA
A03 IS RECOGNIZED BY SEVERAL
T-CELLS, EACH CLONE OF T-CELL
RECEPTOR IS ONE COLOR.
SO THERE'S TWO T-CELLS WITH A
T-CELL RECEPTOR THAT'S THE
SEQUENCE OF THIS BLUE AND
THERE'S THE RED T-CELL RECEPTOR,
THERE'S 14, 10 PLUS FOUR HERE.
THERE'S THESE OTHER TWO ANTIGENS
THAT ARE RECOGNIZED WITH JUST A
COUPLE OF CELLS THAT MADE THE
CUTOFF TO BE ABLE TO RECOGNIZE.
AND WITH NOT ONLY THIS ONE TIME,
WE DID IT OVER TIME, SO NOW I'M
GOING TO ADD THE DIFFERENT TIME
POINTS.
AT BASELINE WE HAD A BIOPSY,
WHICH IS JUST A CORE BIOPSY AND
WE EXPANDED T-CELLS FROM THERE,
SO IT'S A SKEWED READOUT.
HERE'S PBMCs AT DIFFERENT TIME
POINTS, AND THEN WHEN THE
PATIENT WAS RESPONDING, THERE
WAS ANOTHER CORE BIOPSY FROM
THIS LUNG METASTASES ON
EXPANDING THE T-CELLS.
IF WE LOOK AT THIS, AGAIN, IT'S
NON-RANDOM DATA.
IT THE SAME FOUR MUTATED
PEPTIDES, ONE OF THEM
PRESENTED -- THE SAME PEPTIDE
PRESENTED BY TWO HLAs, SO
THERE'S ONLY THREE MUTATIONS
THAT ARE BEING RECOGNIZED OVER
TIME, WHEN WE HAD A LIBRARY OF
243 PUTATIVE HLA NEOANTIGEN
COMPLEXES.
AND THERE'S THESE CLONOTYPES
THAT APPEAR AND DISAPPEAR, SO
THIS ONE IS THE SAME ONE ALL OF
THE TIME, THIS ONE IS THE SAME
ONE ALL THE TIME BUT HERE WE SEE
THIS ANTIGEN HAS MULTIPLE T-CELL
RECEPTORS OF DIFFERENT COLORS
THAT EXPAND AT DIFFERENT TIME
POINTS.
I DON'T THINK THIS TELLS US
ANYTHING ABOUT THE CLONAL
REPOPULATION.
I THINK IT'S SAMPLING ERROR.
WE HAD 10 MLs OF BLOOD IN A
CORE BIOPSY SO THERE'S A LOT OF
SAMPLING ERROR OF THIS BUT WHAT
IT TELLS US IS THAT THE SAME
MUTATIONS KEEP BEING RECOGNIZED
OVER TIME, IN THIS PATIENT
RESPONDING TO ANTIPD-1.
THAT'S PATIENT NUMBER 2, I'LL
SHOW YOU THE SAME GRAPH.
THERE WERE FOUR MUTATIONS,
ACTUALLY THREE MUTATIONS
PRESENTED BY -- ONE OF THEM
PRESENTED BY TWO HLAs AND
21 NEOANTIGEN PCRs.
HERE WE HAVE A LIBRARY OF
243 NEOANTIGEN HLA COMPLEXES ALL
ORGANIZED.
THE ONE IN COLOR ARE THE ONES
THAT HAVE AT LEAST ONE T-CELL
THAT RECOGNIZE IT, SO LET'S SAY
THIS ONE, PLA2GA PRESENTED BY
HLA2.1, THERE'S A SINGLE T-CELL
THAT RECOGNIZES THIS MUTATION AT
ONE TIME POINT, AS OPPOSED TO
THIS NON188, RECOGNIZING IT
MULTIPLE TIMES OR WDR1 THAT HAS
ONLY TWO CLONOTYPES BUT MULTIPLE
T-CELLS EXPANDED IN DIFFERENT
TIMES.
SO HERE THERE WERE 11 MUTATIONS
RECOGNIZED BY 41 DISTINCT T-CELL
RECEPTORS.
SO HOW CAN I CONVINCE THAT THESE
T-CELL RECEPTORS ARE THE RIGHT
ONES AND NOT JUST NOISE OR
BINDING -- NONSPECIFIC BINDING
WHEN WE HAD CELL LINES FROM
THESE PATIENTS, WE CAN GET THESE
T-CELL RECEPTORS, CLONE THEM,
CRISPR EDITING THEM INTO DONOR
T-CELLS AND THEN COCULTURE THEM
WITH THE MELANOMA CELL LINE FROM
THE PATIENT.
SO HERE WE GO BACK TO PATIENT
NUMBER 2, THE ONE THAT HAD TWO
MUTATIONS, PRESENTED BY HLA
2.1 SO T-CELLS EXPRESSING THIS
T-CELL RECEPTOR RECOGNIZING THIS
MUTATION KILLED THE TUMOR, THE
TUMOR IS IN RED, THE T-CELLS
EXPAND AND THAT'S WHY THERE'S
ALL THESE VERY DARK -- THOSE ARE
THE T-CELLS, AND HERE'S A
CONTROL NEOANTIGEN TCR,
ENGINEERED -- WITH CRISPR
EDITING, THE MELANOMA CELLS GROW
PRPROGRESSIVELY.
WE TREAT WITH -- THIS TCR NUMBER
2 KILLED THE TUMOR.
THAT'S ONE OF THE TCRs THAT
I'LL SHOW YOU FROM THAT PLOT BUT
THERE'S 20 OTHERS.
WHAT HAPPENED WITH THE 2 20
OTHERS?
WE DID THE SAME EXPERIMENT AND
ALL OF THEM, ALL 21 OF THEM,
KILLED THE ANTIGEN MATCHED
TUMOR.
TELLING US ASSORTING THESE
CELLS, CLONING THE TCRs AND
GENE EDITING THEM BACK CAN GET
THEM IN SOME CASES AS EFFICIENT
AS THESE ARE, EVERY SINGLE ONE
SELECTED THIS WAY LED TO
CYTOTOXIC ACTIVITY TO THE SAME
CANCER CELL.
THAT'S NOT IN ALL, BUT IT
ACTUALLY WAS THE MAJORITY.
LET'S LOOK AT PATIENT NUMBER ONE
THAT HAD, I THINK, 41 TCRs,
TARGETING 11 MUTATIONS.
NON-NEOANTIGEN -- BY DIFFERENT
HLAs SO HERE ARE ALL OF THE
NEOANTIGENS.
IN THIS CASE IT WAS A LITTLE BIT
DIFFERENT.
THE CELL LINE FROM THIS PATIENT
WERE CALLED MR495, HERE WE'RE
TESTING A SET OF TCRs, ANOTHER
SET OF TCRs, ANOTHER SET OF
TCRs, AND THEY'RE ORGANIZED
LOGICALLY WHERE ALL OF THESE TCR
KILL THE AUTOLOGOUS CELL, NOT
THE MISMATCH.
THIS ONE HAD INTERMEDIATE
KILLING, THESE ONES HAD NO
KILLING AT ALL.
WHEN WE WENT THROUGH THE DATA OF
THIS PATIENT, IT WAS SAID THAT
THE NUP P188 HAD MULTIPLE TCRs
AT MULTIPLE TIME POINTS, THAT
WAS ONE OF THE GOOD KILLERS, AS
OPPOSED TO THE 2GA 4-RBGS THERE
WAS ONLY ONE T-CELL AND THAT WAS
ONLY ONE TIME POINT AND THAT
DIDN'T KILL AT ALL, THEN THERE'S
THE INTERMEDIATE TCRs, ALSO
HAD A PAUCITY OF CLONAL TYPES
RECOGNIZING THESE MUTATIONS.
SO THIS IS PATIENTS WHO HAD
CLINICAL RESPONSES TO ANTI-PD-1
THERAPY THAT MAKES SENSE THAT
THOSE T-CELLS WERE STRONG ENOUGH
TO GET AN IMMUNE RESPONSE THAT
SHRINK THE CANCER AND LED TO
LONG TERM BENEFIT.
SO WHAT HAPPENED IN PATIENTS
WHERE WITH PULL T-CELLS WITH
THIS APPROACH BUT TCRs FROM
PATIENTS THAT DID NOT HAVE
CLINICAL RESPONSE TO THERAPY.
SO WE HAD THREE OF THESE
PATIENTS WHERE WE HAD NO
RESPONSE TO THERAPY, BUT WE GOT
THE CELL LINE AND WE GOT PULLED
TCRs.
WE PULLED A LOT LESS TCRs, SO
FROM PATIENT NUMBER ONE, THERE'S
ONLY ONE CELL THAT RECOGNIZES
ONE MUTATION.
PATIENT NUMBER 9.
PATIENT NUMBER 10, THERE'S THREE
CELLS FROM TWO CLONAL TYPES THAT
RECOGNIZE THE SAME MUTATION.
PATIENT 11, THERE'S TWO CELLS
THAT RECOGNIZE TWO MUTATIONS.
FIVE TCRs ON TOTAL FROM THESE
THREE PATIENTS WITHOUT CLINICAL
RESPONSE TO THERAPY.
SO HOW MANY OF THESE 5TCRs DO
YOU THINK WILL RECOGNIZE AND
KILL THE MATCHED CELL LINE?
ACTUALLY ALL OF THEM.
HERE'S THE TWO TCRs FROM
PATIENT 11, THE ONE ACTUALLY TWO
TCRs FROM PATIENT 10, AND ONE
TCR FROM PATIENT NINE.
ALL OF THEM HAD CYTOTOXICITY TO
THE MATCHED CELL LINE.
SO THESE ARE PATIENTS WHO HAD NO
CLINICAL RESPONSE TO ANTI-PD-1
BUT WE CAN STILL PULL RARE
T-CELLS FROM BLOOD OR THE TUMOR
AND ISOLATE BASED ON THEIR
ANTIGEN SPECIFICITY AND GENE
ENGINEER THEM AND SHOW THIS
ACTIVITY.
SO FROM THIS PART OF THE TALK,
WE CAN CONCLUDE THAT NEOANTIJET
SPECIFIC T-CELLS CAN BE
IDENTIFIED FROM PATIENTS WITH
METASTATIC MELANOMA THAT HAVE OR
DO NOT HAVE CLINICAL RESPONSES
ENDUED BY PD-1 BLOCKADE-BASED
THERAPY.
BUT THESE NEOANTIGEN-SPECIFIC
TCRs WHEN -- ONE STEP OF
CYTOSPECIFIC JEAN ENGINEERING
WHERE WE KNOCK OUT ENDOGENOUS
T-CELL RECEPTOR ALPHA BETA --
THAT WE CAN GET THEM TO HAVE
REACTIVITY AGAINST MELANOMA
CELLS.
AND THAT THIS IS THE -- THESE
NEOANTIGEN-SPECIFIC T-CELL
RESPONSES ARE NON-RANDOM,
THEY'RE NOT MORE MUTATIONS, MORE
T-CELLS ARE RECOGNIZED IN MORE
MUTATIONS, BUT THERE'S
IMMUNODOMINANCE INTO THE
RECOGNITION, BUT TO THOSE
IMMUNODOMINANT MUTATIONS
PRESENTED BY -- HLA, THE
CLINICAL RESPONSE, PATIENTS WITH
CLINICAL RESPONSE HAVE A
DIVERSITY OF CLONOTYPES
RECOGNIZED IN THOSE FEW
MUTATIONS.
WHICH IS ALL GREAT DESCRIPTIVE
SCIENCE, I THINK, BUT WHAT DO WE
DO WITH IT?
WHAT WE DO WITH IT IS WE WANT TO
TAKE THESE AND MAKE THEM INTO
THERAPEUTIC BECAUSE THOSE T-CELL
RECEPTORS FROM PATIENTS WHO
DON'T RESPOND TO ANTIPD-1 CAN
STILL KILL IN VITRO, WE DON'T WE
DO THIS IN HUMANS? WE ALL KNOW
WHEN WE DO THINGS IN HUMANS, IT
TAKES A LITTLE MORE EFFORT BUT
AT LEAST WE MADE AN ATTEMPT AND
THIS IS THE SECOND PART OF THIS
PRESENTATION THAT WAS
PRESENTED -- THAT WAS MOST OF
THE DATA PRESENTED BY STEPHANIE
MANDELL AND WE HAVE AN ARTICLE
ALREADY IN NATURE THAT SOME OF
YOU HAVE COMMENTED ON.
HERE WE'RE DOING THE SAME PART
WE DID BEFORE, THE TUMOR
SEQUENCING COMPARED TO BLOT,
MAKE THE LIBRARY OF THE HLA
PEPTIDES WITH THE SINGLE CHAIN
TRIMERS, MULTI-MERRIZED TO
T-CELLS FROM BLOOD IN THIS CASE
AND CLONE THE T-CELL RECEPTORS
FROM SINGLE CELLS AND THEN
PRODUCE T-CELLS BY GENE
ENGINEERING, BY THIS CRISPR
BASE -- OF CAS9 GUIDE RNA AND ON
THE PEPTIDE -- AND THEN WE DID
THIS FOR PATIENTS, WE DID UP TO
THREE T-CELL RECEPTOR PER
PATIENT BECAUSE FROM THE START,
WE DON'T WANT TO DO ONE AND HAVE
THE TUMOR ESCAPE TO ONE T-CELL
RECEPTOR SO WE MADE IT EVEN
HARDER FOR OUR CELLS AND START
WITH THE T-CELL RECEPTOR
PRODUCT.
IF WE LOOK AT FROM THE PATIENT
PERSPECTIVE, PATIENTS WHO WERE
ENROLLED TO LOOK FOR ARCHIVAL
TISSUE TO DO THE SEQUENCING, AND
THEN ONCE WE HAD THE T-CELL
RECEPTORS SH PATIENTS, PATIENTS,
TESTING, THEN THE PATIENTS WOULD
COME IN, CHEMOTHERAPY, THE THREE
T-CELL RECEPTOR AND HAVE AN
EARLY BIOPSY, AND THEN A LOOK AT
ANY POTENTIAL OF TUMOR
RESPONSES.
IT WAS A DOSE ESCALATION PHASE
ONE TRIAL.
WE STARTED ON -- GOING UP TO
FOUR TIMES 10 TO THE 9.
AND THEN THIS PROTOCOL, DESPITE
MY COMPLAINING, DIDN'T HAVE
IL2 AS A COMPONENT FROM THE
START BUT THEY WERE COHORTS THAT
WERE ALLOWED TO GIVE
IL2 AFTERWARDS.
WE ENROLLED 189 PATIENTS, AND WE
LOOKED FOR T-CELL REACCEPT PO RM
89 PATIENTS.
FROM THE FIRST 88 PATIENTS WITH
A LIBRARY OF 64 HLAs, WE
PREDICTED OVER 50,000
NEOANTIGENS, AND HERE THEY'RE
ORGANIZED BY LOW AFFINITY TO
HIGH AFFINITY.
EACH DOT WOULD BE PREDICTED
NEOANTIGEN PRESENTED BY ONE OF
THE HLAs OF THAT PATIENT.
THAT LED TO PACPHARMA MAKING
12,000 DISTINCT NEW ANTIGEN
REACTIONS, COCULTURED WITH THE
T-CELLS.
WE CAN SEE THAT THESE ARE THE
GREEN DOTS AND IT'S SKEWED OVER
HERE BECAUSE ONLY THE HIGHER
AFFINITY PEPTIDES WOULD
STABILIZE THE SINGLE CHAIN
TRIMER, IF IT'S LOW AFFINITY, IT
FALSE OFF.
AND THEN HOW MANY T-CELL
RECEPTORS WERE PULLED, THERE ARE
OVER 900 UNIQUE NEOANTIGEN
RECEPTORS, PROBABLY THE HIGHEST
SO FAR THAT ANYBODY HAS CLONED,
COULD BE ISOLATED FROM ALL OF
THESE SAMPLES.
AGAIN, THERE'S A SKEWING TO THE
HIGHER AFFINITY, BUT THERE'S
ALSO LOWER AFFINITY T-CELL
RECEPTORS THAT WOULD DISTINCTLY
BIND WITH ALL OF THE CUTOFFS
THAT WE HAD TO GET OUT FROM
BACKGROUND.
THE DATA FROM THESE PATIENTS IS
LIKE YOU SAW BEFORE, WITH THE
PATIENTS WITH MELANOMA, SO HERE
IS A LIBRARY OF 26 262 PIP TIED
LIBRARY COVERING MUTATIONS FROM
LOW AFFINITY TO HIGH AFFINITY.
THERE'S ONLY ONE, TWO, THREE AND
FOUR MUTATIONS BEING RECOGNIZED,
THIS ONE PRESENTED BY THREE OF
THE HLA -- SO THIS MUST BE TWO
MUTATIONS, I'M SORRY, AND THIS
IS ONE OF THEM.
HERE THERE'S MULTIPLE -- I'M
SORRY IT'S NOT LABELED
COMPLETELY WITH THE MUTATIONS
BUT IN THE PAPER, THERE'S A
TABLE WITH ALL OF THE HLAs,
THE SEQUENCES OF THE MUTATIONS
AND THE SEQUENCES OF THE T-CELL
RECEPTORS, IF ANYBODY IS
INTERESTED ON THAT.
FROM CLONING THE T-CELL
RECEPTORS, THEY HAVE TO BE GENE
EDITED AND THIS IS -- THESE
PRECISION CAS9-BASED GENE
EDITING KNOCKING OUT THE T-CELL
RECEPTOR ALPHA AND BETA, AND
THEN KNOCKING INTO THE T-CELL
RECEPTOR ALPHA, THE TWO -- THIS
PEPTIDE THAT HAS -- THE PLASMID
THAT HAS THE TRANSGENIC TCR
ALPHA AND PART OF THE BETA --
THE BETA AND PART OF THE ALPHA,
AND THEN EXPAND THE CELLS AND
GIVE THEM TO PATIENTS.
HERE'S THE SCHEMATIC OF THE
PEPTIDE, SO IT'S THE FULL
SEQUENCE TCR BETA WITH 2A AND
THE VARIABLE PART OF THE TCR
ALPHA BECAUSE WE'RE PUTTING
UNDER TRACK LOCUS DOUBLE LATE TO
EXPRESSION OF THE TRANSGENIC TCR
ALPHA AND BETA.
THAT WENT INTO MANUFACTURING OF
THE T-CELLS, AND WITH TIME, AS
WE'RE RUNNING THE CLINICAL
TRIAL, THE MANUFACTURING KEEPING
PROBING, TO THIS IS THE VERSION
1.1 ONLY HAD 10 KNOCK-IN -- 10%
KNOCK-IN AND VARIABLE KNOCK-OUT,
VERSION TWO HAD INCREASED AND
VERSION THREE WENT UP TO 90%
KNOCKOUT AND 40% KNOCK-IN WITH
THE CRISPR BASE, WHICH IS
GETTING CLOSE TO WHAT YOU CAN
GET WITH A RETROVIRUS.
AND WITH THE NUMBER OF CELLS
PRODUCED WITH THE VERSION THREE
WAS ALREADY AT THE LEVELS WE
COULD GO UP IN THE DOSE
ESCALATION.
I'LL SHOW YOU THE DETAILS OF THE
GENE EDITING.
SO HERE IS CD8 T-CELLS AND
CD4s.
HERE'S UNEDITED CELLS, AND
HERE'S WITH THE T-CELL RECEPTOR.
THIS PATIENT RECEIVED T-CELL --
THREE T-CELL RECEPTORS, 1036,
1033, AND 1037.
THIS FIRST T-CELL RECEPTOR IS ON
CD8s AND CD4s, SO IT'S A
CD8 -- TRANSGENIC T-CELL
RECEPTOR.
THESE TWO OTHER T-CELL RECEPTORS
ARE ONLY EXPRESSED IN CD #s
CD8s ANDNOT IN CD4s SO THEY ARE
CD8 DEPENDENT SO THEY NEED THE
CORE RECEPTOR TO BE EXPRESSED ON
THIS SURFACE AND THEN BIND TO
THE COMPLEXES THAT THE
MULTI-MERES THAT WE HAVE THAT WE
USE FOR SELECTING AND ALSO WE
USE FOR VALIDATION.
NOT ONLY WE VALIDATED THE
EXPRESSION BUT THE POTENCY ASSAY
WAS IF WE GIVE THESE SOLUBLE HLA
PEPTIDE COMPLEXES THAT WE CALL
COMPACTS, WOULD THAT INDUCE
PROLIFERATION OF DONOR T-CELLS
THAT EXPRESS THE T-CELL RECEPTOR
THAT WAS PART OF THE TESTING.
WE CAN SEE THAT THE GENE
EDITED -- THEY ALL HAVE DILUTION
OF CFSC SHOWING PROLIFERATION.
THE CELLS THAT HAVE ONLY
KNOCKOUT AND NO -- DO NOT
PROLIFERATE AND THE WILD TYPE
CELLS WITHOUT THE KNOCK-OUT OR
KNOCK-IN DO NOT PROLIFERATE SO
IT'S TRUANT GENERAL-SPECIFIC
PROLIFERATION.
SO GOING BACK TO THE PATIENTS,
WE ENROLLED 187 PATIENTS THAT
WENT THROUGH DIFFERENT PHASES OF
THE SCREENING.
88 PATIENTS HAD ENOUGH OF THE
TISSUE AND DATA THAT WE COULD DO
THE SCREENING OF T-CELL
RECEPTORS.
THAT'S THE DATA I SHOWED YOU
BEFORE.
28 MADE TO LOU KA PHORESIS AND
16 PATIENTS WERE DOSED OVER A
PERIOD OF TIME.
THESE 16 PATIENTS HAD DIFFERENT
CANCERS.
THERE WAS NO RESTRICTION -- THIS
WAS A PHASE ONE TRIAL THAT
ALLOWED PATIENTS WITH MULTIPLE
CANCERS TO COME IN.
IT TURNED OUT THAT ONE SIDE PUT
A LOT OF PATIENTS WITH
COLORECTAL CANCER AND THOSE
COVERED THE FIRST COHORT, SO
THAT WAS THE PREDOMINANT NUMBER
OF PATIENTS THAT WERE INVOLVED.
THIS IS A SLIDE THAT'S PAINFUL
BECAUSE IT SHOWS THE TIME THAT
IT TOOK FROM BASELINE TO GETTING
THE INFUSION IN THESE 16
PATIENTS.
AND HERE WE HAVE ONE YEAR.
SO MAJORITY OF THE PATIENTS TOOK
UP TO A YEAR.
BUT IT WAS NOT THAT ONE STEP WAS
MORE LENGTHY.
HERE IS COLOR-CODED INFORMED
CONSENT, BIOINFORMA TICKETS,
IMPACT ISOLATION, YOU CAN SEE
THE GREY BARS ARE LONG SO JUST
GETTING THE PATIENTS IN AND
START THE PROCESS TOOK QUITE A
WHILE AT MULTIPLE SITES.
THESE ORANGE ONE MAY TAKE LONG
SO THE ISOLATION OF THE --
MAKING THE HLA COMPLEXES, BUT
ONCE WE HAD THE CELL
MANUFACTURING TO DOSING, IT ALL
BECAME MUCH SHORTER.
SO IT'S -- THE LATER PART WAS
MORE CONTROLLABLE.
BUT WHY THAT?
WELL, IF WE 50 BACK GO BACK TO S
CARTOON, WE SCREENED
187 PATIENTS, WE TREATED 16, AND
IN BETWEEN, WE MADE THE MEDIAN
OF 35 -- WE TARGETED A MEDIAN OF
35, RANGE OF 20 TO 236 EXPRESS
MUTATIONS PER PATIENT.
A MEDIAN OF 104 PEPTIDE HLA
LIBRARIES WERE MADE.
THE MAJORITY WENT FROM 49 TO 26F
1,841 PEPTIDE-HLA PROTEINS WERE
PRODUCED TO TREAT THESE 16
PATIENTS WITH A MEDIAN OF SIX
HLAs TARGET PER PATIENT SO
COVERING MOST OF THE HLA
CLASS I.
WE HAD THE MEDIAN OF EIGHT
NEOTCRs FOR PATIENT FOR A
TOTAL OF 175 TCRs IN THE GROUP
OF 16 PATIENTS, AND AT 37
NEOANTIGEN TCRs WERE INFUSED
BACK TO THE 16 PATIENT.
THEY HAD DIFFERENT EC50s FROM
LOW AFFINITY TO VERY HIGH
AFFINITY.
HERE'S THE CLINICAL RESULTS.
THERE WERE NO OBJECTIVE
RESPONSES, FIVE PATIENTS WITH
STABLE DISEASE.
SOME OF THEM, IT COULD BE FROM
THE CONDITIONING CHEMOTHERAPY.
I'LL SHOW YOU THE ONE PATIENT WE
THINK THAT WAS NOT THE
CONDITIONING CHEMOTHERAPY.
I'M SORRY, I'LL GO BACK.
AND THE TOXICITIES WERE MOSTLY
THAT THE CONDITIONING THERAPY,
THERE WAS ONE PATIENT WITH
HISTORY OF BRAIN METASTASES THAT
HAD SOME KIND OF ENCEPHALOPATHY
FOR ONE DAY AND WAS GIVEN
CORTICOSTEROIDS.
WE'LL NEVER BE SURE WHAT THAT
WAS, BUT IT SEEMED TO BE THAT
THIS NEOANTIGEN T-CELLS, THE
MAJORITY OF THEM, PATIENTS HAD
NO DISCERNING TOXICITIES EVEN
THOUGH WE CLONED THEM IN LARGE
NUMBERS OF THEM.
SIMILAR TO YOUR EXPERIENCE WITH
TIL CULTURES.
SO HERE ARE THE THREE DOSE
LEVELS.
DOSE LEVEL ONE, DOSE LEVEL TWO,
DOSE LEVEL THREE.
DOSE LEVEL ONE, THERE WAS VERY
LOW EXPOSURE, SO VERY FEW
T-CELLS EXPANDED AND STAYED
AROUND.
THAT WAS WITH LOW DOSES OF
CONDITIONING CHEMOTHERAPY AND
WITHOUT IL-2.
DOSE LEVEL TWO WAS HIGHER DOSES
BUT ALSO WE CHANGED THE
CONDITIONING BECAUSE WE THOUGHT
THERE WAS NO -- NOT ENOUGH
LYMPHODEPLETION AND THAT LED TO
MORE EXPOSURE.
THOSE LEVEL THREE, SOME PATIENTS
REALLY HAD GOOD EXPOSURE LIKE
THIS LAST ONE, WHERE 40% OF THE
T-CELLS AT ONE POINT CIRCULATING
IN BLOOD WHERE ANTIGEN -- WHERE
TCR TRANSGENIC T-CELLS.
BUT HERE WE HAVE HIGH EXPOSURE.
WITH IL-2, THERE SEEMS TO BE
HIGH EXPANSION INITIALLY THAN
THE ONES WITHOUT THE IL-2.
THE KEY POINT OF ALL OF THIS IS
DID WE REDISTRICT IMMUNE SYSTEM
TO THE CANCER BECAUSE THERE WERE
THE MUTATIONS ALLOWED THE
T-CELLS TO GET THEM -- TO GET
THOSE T-CELL RECEPTORS TO
RECOGNIZE CANCER CELLS.
AND HERE WE HAVE A SERIES OF
PATIENTS ON DOSE LEVEL ONE, DOSE
LEVEL TWO AND DOSE LEVEL THREE
WITH A BASELINE AND A
NON-THERAPY BIOPSY AND THIS
PATIENT RECEIVED THREE T-CELL
RECEPTORS, ONLY ONE WAS
RECOGNIZED AT BASELINE.
THIS IS A LOGARITHMIC SCALE ON
THE Y AXIS.
YOU CAN SEE THAT THE THREE
T-CELL RECEPTORS NOW BECAME
PRESENT IN THE TE TUMOR.
HERE THERE'S 1 T-CELL RECEPTOR
THAT SPANNED HERE, THREE T-CELL
RECEPTORS EXPANDED AND HERE YOU
CAN SEE THE DATA FROM ALL OF
THESE PLOTS WOULD COME SO
PATIENT 612, SO THIS ONE HERE
FROM DOSE LEVEL THREE, AT
BASELINE WE COULD NOT DETECT ANY
OF THE T-CELL RECEPTORS AMONG
THE MOST EXPRESSED T-CELL
RECEPTORS, AND POST DOSING,
THOSE THREE T-CELL RECEPTORS
WERE 6%, 1.4% AND 1.1% OF THE
T-CELL RECEPTORS THAT WERE
PRESENT IN THE TUMOR.
TELLING US THAT WE COULD
EFFICIENTLY REDIRECT T-CELLS TO
TUMORS BY GIVING THEM THESE
TRANSGENIC T-CELL RECEPTORS.
THIS ONE PATIENT THAT WE THINK
THAT WAS LITTLE EVIDENCE OF
RESPONSE BUT SOME IS A PATIENT
WHO HAD RECEIVED -- WHO HAD
LUNG -- METASTATIC LUNG CANCER
WITH SIX -- LINES OF THERAPY AT
THE TIME OF SCREENING, WENT UP
TO NINE AT THE TIME OF DOSING,
HAD ONLY ONE T-CELL RECEPTOR
PRODUCT AND HAD IL-2 FOR 6 DAYS
AND WE CAN SEE TWO OF THE
METASTASES RETROPERITONEAL AND
OVARIAN MASS THAT SLIGHTLY
DECREASED IN SIZE.
OTHERS INCREASED IN SIZE.
SO THIS IS -- I'M NOT SAYING
THIS IS THERAPEUTIC BUT AT LEAST
THERE WAS A HINT THAT THESE
T-CELL RECEPTORS, EVEN THOUGH
THEY WERE PROBABLY TOO FEW AND
TOO LATE FOR THIS PATIENT, HAD
SOME EFFECT.
AND IF YOU WOULD SAY, WELL, WHY
DEPARTMENT YOU HAVE BETTER
RESPONSES, THESE T-CELL
RECEPTORS KILL CANCER CELLS IN
VITRO, WHY DIDN'T YOU GET BETTER
RESPONSES IN THIS CLINICAL
TRIAL?
WELL, WE'VE DONE A WHOLE BUNCH
OF STUDIES GOING BACK, SO HERE
IS LOOKING AT THE POTENCY OF THE
T-CELL RECEPTORS, HERE THE 37
T-CELL RECEPTORS GIVEN TO
PATIENTS IN THE SEMI LOGARITHMIC
SCALE AND HERE ARE T-CELL
RECEPTORS THAT MANY OF YOU GUYS
HAVE DEVELOPED, THE DMF5, WHICH
IS A HIGH AFFINITY T-CELL
RECEPTOR HERE, AND A PAUL
ROBBINS ENGINEERED T-CELL
RECEPTOR IS HERE, THEY'RE ALL
HIGHER AFFINITY THAN THE MEDIAN
OF THE PAC T-CELL RE ACCE ACCEPS
AT LEAST WITH THESE PATIENTS
WITH EPITHELIAL CANCERS.
NOT ONLY -- BUT THE EXPRESSION
OF THE ANTIGEN, WE CAN SEE THAT
THE PACKED NEOANTIGENS WERE IN
MEDIAN LOW EXPRESSION LIKE KRAS,
AS EXPOSED TO HIGHLY EXPRESSED
TARGETS IN THE CANCER CELLS THAT
MAKE IT BETTER TARGETS FOR
T-CELLS.
THE LAST THING IS, ARE WE DOING
THIS TOO LATE AND TOO LITTLE.
AND WE LOOKED FOR HLA LAWS OF
HETEROZYGOSITY RETROSPECTIVELY
AND THESE ARE ALL OF THE
BIOPSIES WE LOOKED FOR THESE
DIFFERENT PATIENTS.
WHEN WE SEE AN HLA WITH GREEN,
THAT MEANS THIS T-CELL RECEPTOR
RECOGNIZED A B ALLELE, AND THAT
B ALLELE WAS EXPRESSED.
BUT IF THERE'S A RED, IT MEANS
THAT THIS T-CELL RECEPTOR FROM
THE PATIENT THAT RECEIVED THE
THREE T-CELL RECEPTOR PRODUCT
HAD LOSS OF HETEROZYGOSITY.
SO WE GAVE TO THIS PATIENT ONE
T-CELL RECEPTOR THAT THE TUMOR
NO LONGER EXPRESSED BECAUSE OF
LOSS OF HETEROZYGOSITY SO THAT
HAPPENED IN ONE, TWO, THREE
PATIENTS, THIS ONE HAD TWO FROM
THE C ALLELE THAT HAD LOST
EXPRESSION, SO THE PEPTIDE WAS
NO LONGER EXPRESSED BY THE TIME
WE HAD DONE ALL OF THESE AND
MADE IT INTO THE CLINIC.
SO WHAT I WANT TO CONVEY WITH
THIS IS THAT ACTUALLY WE CAN DO
IT.
I'M NOT SAYING THAT IT'S EASY OR
THAT IT'S GOING TO BE
THERAPEUTIC ANY TIME SOON, BUT
WE CAN ISOLATE AND CLONE T-CELL
RECEPTORS THAT CAN RECOGNIZE
PATIENT-SPECIFIC MUTATIONAL KNEW
OWE AT GENERALS, WE CAN USE THIS
ONE STEP SINGLE NON-VIRAL PREER
SITION GENE EDITING TO INSERT
THE KNOCK-IN THE T-CELL RECEPTOR
AND KNOCK OUT THE ENDOGENOUS
T-CELL RECEPTORS, THAT WE CAN DO
THIS IN -- AND MAKE UP TO
THREE -- PER PATIENTS, WE CAN
SAFELY REINFUSE THEM TO PATIENTS
AND WE CAN DEMONSTRATE THAT THAT
REDIRECTS T-CELLS TO ACCUMULATE
IN THE TUMOR BECAUSE AFTERWARDS
THOSE T-CELL CLONOTYPES THAT --
TCRs WERE THE MOST EXPRESSED
OF THE TCRs IN THE TUMORS.
AND RETROSPECTIVE ANALYSIS TELLS
US SOME PLACES WE HAVE TO
IMPROVE, WE SHOULD GET THE
HIGHER AFFINITY T-CELL
RECEPTORS, HIGH EXPRESSION OF
THE ANTIGENS, AND FOR SURE MAKE
SURE THAT THAT TUMOR BY THE TIME
WE TREAT THE PATIENT HAS NOT
DONE IMMUNOEDITING APPROACHES
THAT -- EXPRESSION.
THE DATA I PRESENTED TO YOU IS
WORK FROM MANY PEOPLE.
PROBABLY THE PAC LIST SHOULD GO
TO 200 PEOPLE.
THESE ARE THE PEOPLE FROM MY
LAB.
I TALKED ABOUT CRIS PUIG-SAUS,
NOW FACULTY MEMBER AT UCLA
PROCEEDING TO THE
CHARACTERIZATION OF THESE
NEOANTIGEN SPECIFIC TCRs THAT
WE DESCRIBED.
ELENA GROSS TRAINED IN SPAIN,
HELPED US A LOT IN SETTING UP
THE FIRST CULTURES, THE
TECHNOLOGIES WERE DEVELOPED WITH
JIM KEITH AND DAVID BALTIMORE,
WITH MULTIPLE FUNDING SOURCES,
NIH -- SO I'D BE HAPPY TO TAKE
ANY QUESTIONS OR COMMENTS.
[APPLAUSE]
>>HI.
THIS IS A TOUR DEFORCE.
THANK YOU.
I THINK IT'S IMPORTANT FOR
EVERYONE HERE TO SEE THIS TYPE
OF STUDY.
HOW MUCH OF THE ACTIVITY OR LACK
OF ACTIVITY DO YOU THINK WAS DUE
TO IMMUNOSUPPRESSIVE FACTORS IN
THE TUMOR MICROENVIRONMENT?
THAT IS, WERE YOU ABLE TO
ANALYZE ANYTHING IN THE TUMOR
MICROENVIRONMENT IN TERMS OF
STABLE DISEASE VERSUS
PROGRESSIVE DISEASE?
>>WELL, WE LOOKED AT
TRANSCRIPTOME ANALYSIS OF THE
TUMORS BECAUSE WE HAVE IT IN ALL
OF THEM AND THE COLORECTAL
CANCERS, THE MAJORITY OF THEM
WERE THESE CS4 THAT SEEMS TO BE
THE MOST IMMUNE SUPPRESSIVE
WHERE THERE'S NATURAL LESS
T-CELLS AND THOSE WERE THE ONES
THAT ARE ACTUALLY MORE COMMON
THAT DON'T RESPOND TO ALL THE
THERAPIES.
I DON'T THINK THE TUMOR
MICROENVIRONMENT IS WHAT LED TO
NO RESPONSE, I THINK NOT HAVING
ENOUGH T-CELLS AND GIVING ENOUGH
OF THEM.
THIS WAS LIKE BUILDING A PLANE
AS WE'RE FLYING IT, AND THE
MANUFACTURING IMPROVED.
THE LAST COHORT OF PATIENTS ARE
THE ONES THAT HAVE ENOUGH
EXPANSION OF T-CELLS AND ENOUGH
NUMBERS THAT COULD HAVE A
POTENTIAL TO WORK, AND THAT'S
THE ONLY TIME WHERE WE SAW SOME
ACTIVITY.
SO WE CANNOT -- I CANNOT ANSWER
YOUR QUESTION BECAUSE I DON'T
THINK WE HAVE TREATED ENOUGH
PATIENTS WITH THE FULL THERAPY
TO BE ABLE TO ANSWER IT.
>>THANK YOU.
>>THANK YOU, THAT WAS VERY,
VERY INTERESTING.
ACTUALLY IT'S NOT THAT MANY
PEOPLE SO I HAVE TWO QUESTIONS.
SO THE FIRST QUESTION I HAVE IS,
THERE'S SO MANY ANTIGENS,
THERE'S SO MANY EPITOPES, AND
YOU IDENTIFY THEM ALL, YOU HAVE
THIS ONE MAP AND THEN -- WHY ARE
THERE SUCH FEW TCRs FINALLY
RECOGNIZING THIS?
>>SO THIS CONCEPT OF
IMMUNODOMINANCE IS SOMETHING
THAT HAS EXISTED IN THE VIRAL
LITERATURE FOR YEARS, WHERE IF
YOU TAKE AN ANTIGENIC PROTEIN
AND IMMUNIZE A MOUSE OR A HUMAN,
IT TENDS TO FOCUS -- AN
ANTIGENIC VIRUS, IT TENDS TO
FOCUS ON VERY FEW PROTEINS.
I REMEMBER STUDIES FROM ELI
SARCARS ON AUTOIMMUNITY WHERE IF
YOU TOOK SOME OF THE MORE
IMMUNOGENIC PROTEIN OUT THEN
THERE WERE DOM NAPT CLONES THATT
WOULD BE RECOGNIZED.
I DON'T KNOW THE BASIS OF
IMMUNODOMINANCE, THAT'S THE
DESCRIPTIVE PART OF HIS WORKT.
WE TRIED TO COVER AS MUCH AS
POSSIBLE.
WE KNOW WE HAVE VERY LOW
BACKGROUND BECAUSE THE WAY WE
DID THE SORTING OF THE CELLS
TOOK OUT MOST OF THE --
FLUORESCENCE, IT WOULD MINIMIZE
BACKGROUND.
THE POSITIVE HITS ARE HITS, BUT
THEY ALL MAKE LET'S -- MUTATION
AND HLAs, THE MECHANISM, I
DON'T KNOW.
>>SO THE OTHER QUESTION IS, YOU
KNOW, YOU SHOW IN THE END THE
DOWN REGULATION OF THE HLA AND
THAT'S WHY THE TUMOR WAS NOT
RECOGNIZED IN SOME CASES.
SO THAT BRINGS UP A QUESTION,
AND YOU SAY THAT IT TAKES -- IT
TOOK YOU A YEAR TO DEVELOP THIS.
SO DOES THIS SUGGEST THAT IT'S
BETTER TO TREAT A PATIENT THAT
WOULD ACTUALLY RESPOND TO AN
IMMUNE CHECKPOINT INHIBITOR, OR
ONE THAT ACTUALLY POTENTIALLY
DOES NOT RESPOND BECAUSE IT'S
LESS LIKELY TO DEVELOP DOWN
REGULATION OR, YOU KNOW, ANY
OTHER IMMUNE ESCAPE?
>>SO THE MORE IMMUNE EDITING
WOULD BE -- CANCER IS MORE
IMMUNOGENIC, ESPECIALLY IF
YOU'RE DOING IMMUNOTHERAPY AND
THE TUMOR BECOMES RESISTANT,
THAT'S WHEN WE SEE GENETIC --
MORE FREQUENT GENETIC MECHANISMS
OF RESISTANCE AS OPPOSED TO
BASELINE, SO IT COULD BE HLA
LOH, IT COULD BE B2M HOMOZYGOUS
LOSS OR JAK1 OR JAK2 MUTATIONS
SO THOSE PATIENTS WOULD NOT BE
TREATED WITH THESE AND THE
ALGORITHMS, THE BIOINFORMATIC
ALGORITHMS ARE NOW ABLE TO
DETECT THAT.
IT'S JUST THAT IT NEEDS TO BE ON
A RECENT BIOPSY, NOT THE
BASELINE BIOPSY FROM A LONG TIME
AGO.
THE LONG TIME, OBVIOUSLY AS
WE'RE GOING IN, EVERYTHING CAN
BE IMPROVED.
IF YOU PUT 10 PEOPLE MORE TO
WORK ON EVERY STEP, IT WOULD GO
MUCH FASTER.
SO IT'S WHAT COULD BE VISIBLY
DONE WITH A STARTUP COMPANY.
>>THANK YOU.
>>SO TONY, JUST WONDERFUL WORK,
AND THANKS SO MUCH FOR SHARING
WITH US.
I JUST HAD A QUICK QUESTION
ABOUT THE -- HOW YOU SCREENED
FOR THE ON TARGET OFF TUMOR
POTENTIAL FOR THE TCRs BEFORE
YOU PUT THAT INTO PATIENTS.
>>THAT WAS NOT -- THAT WAS NOT
PART OF THE PROCESS.
SO THE WHOLE PREMISES THAT THIS
IS AUTOLOGOUS.
>>YEAH.
>>WE WERE ABLE TO GET THROUGH
THE FDA REVIEW AND DO THIS IN
PATIENTS.
AND THERE'S SOME THINGS WHERE
THE FDA COULD SAY YOU'RE NOW
TAKING A TCR, YOU'RE ENGINEERING
INTO ALL THE T-CELLS EU GIVING
IT BACK, WHAT IF THAT TCR
RECOGNIZED THE NON-MU TAITIVE
PEPTIDE?
WELL, THAT WAS NOT PART OF THE
CRITERIA.
BUT THAT TCR ALREADY CAME FROM
THE PATIENT, IT WAS ALREADY
CIRCULATING.
>>RIGHT.
>>SO THE CHANCES OF INDUCED
AUTOIMMUNITY FROM THAT WOULD BE
LOWER, THAT'S NOT IMPOSSIBLE.
MY BIGGEST ARGUMENT FOR ANYBODY
WHO COULD SAY, OH, YOU SHOULDN'T
BE DOING THIS, IS THE TIL
THERAPY.
SO YOU GUYS HAVE BEEN GIVING TIL
TO PATIENTS FOR YEARS, HAS THERE
EVER BEEN A PATIENT WHERE YOU
INDUCE AUTOIMMUNITY OTHER THAN
VITILIGO WITH IMMUNE THERAPY?
I DON'T RECALL IT.
SO EVEN T-CELLS THAT MAY
RECOGNIZE SELF ANTIGEN, IF YOU
GIVE THEM IN LARGE DOSES OR
ADOPTIVE CELL TRANSFER DO NOT
BECOME -- AUTO REACT.
>>PERFECT.
THANK YOU.
>>PAUL OR TIM?
>>YOU SHOW ONE SLIDE WHICH WAS
LOOKING, COMPARING THE AVIDITY
OF THE T-CELLS, I BELIEVE, FROM
EFFECTIVE TCRs IN PREVIOUS
TRIALS AND YOUR TCRs.
I KIND OF PERHAPS APOLOGIZE FOR
NOT UNDERSTANDING THAT
COMPLETELY, BUT WERE YOUR TCRs
OF LOWER AVIDITY SO THAT WAS A
POTENTIAL PROBLEM, DO YOU HAVE
AN IDEA OF PERHAPS HOW TO
OVERCOME THAT ISSUE?
>>WHEN WE STARTED THE PROTOCOL,
SO IF A PATIENT -- WITH ALL OF
THE APPROACH LED TO 10 TCRs
AND WE HAD TO CHOOSE THE THREE
BEST ONES, THEY SET UP A SERIES
OF CRITERIA, YOU WANT TO HAVE
THE THREE TCRs STARTED IN
DIFFERENT HLAs, YOU WANT TO
HAVE THE THREE TCRs TARGETED
IN DIFFERENT ANTIGENS IF YOU CAN
CHOOSE, AND THEN AVIDITY WAS
SOMETHING THAT AT THE BEGINNING
WAS NOT PART OF THIS.
THAT WAS BECAUSE THERE'S THIS
DATA IN THE LITERATURE SAYING
THAT LOW AVID IITY T-CELLS MAY E
ABLE TO STAND MULTIPLE ROUNDS OF
ANTIGEN EXPOSURE AS OPPOSED TO
HIGH AVIDITY THAT WOULD BE
TURNED OFF EARLIER.
IN RETROSPECT, WE SEE THE ONES
THAT EXPANDED MORE IN THE TUMOR,
I DIDN'T SHOW THAT, BUT THE DATA
IS THERE, WERE THE HIGHER
AVIDITY T-CELL RECEPTORS AND THE
ONES THAT EXPANDED LESS IN THE
TUMORS WERE THE LOWER.
SO WE HAVE TO DO THIS AGAIN, WE
WOULD PUT THE AVIDITY AS ONE OF
THE CRITERIA TO SELECT IF
POSSIBLE.
IN SOME PATIENT, THERE WERE ONLY
ONE OR TWO TCRs THAT WERE
ISOLATED THAT WERE JUST USED.
>>THANK YOU.
>>BEAUTIFUL TALK.
>>THANKS.
>>SO WHEN YOU SHOWED THAT THE
T-CELL REPERTOIRE, THE TCR
REPERTOIRE WAS HIGHER IN THE
RESPONDERS, CAN YOU SPECULATE ON
WHAT DO YOU THINK THAT MEANS?
WHY WOULD THAT GIVE YOU A BETTER
CHANCE OF RESPONDING?
WHAT DO YOU THINK IS RESPONSIBLE
FOR THAT?
>>SO WHATEVER MAKES A MUTATION
YOU ME KNOWGENIC IN THOSE
PATIENTS WHO GO ON TO RESPOND TO
ANTI-PD-1 HAVE MORE OF THOSE
MUTATIONS -- DO NOT HAVE MORE OF
THOSE MUTATIONS BUT HAVE MORE
T-CELLS THAT RECOGNIZE THE
MUTATION WITH NONE OR MULTIPLE
TCRs.
>>SO IT'S JUST A NUMBERS GAME,
YOU THINK?
>>I THINK IT'S AN INDIRECT
MEASURE OF HOW GOOD THAT
MUTATION AND HLA COMPLEX WAS TO
TURN ON THE T-CELLS OF THAT
PATIENT.
AND IT'S NOT LINEAL LIQUOR LALYD
WITH THE MUTATIONAL LOAD, THAT'S
WHY IN -- MUTATIONS, RESPONSE TO
ANTI -- IS NON-LINEARLY
CORRELATED.
>>DO YOU THINK THESE STUDIES
WILL CONTINUE THERE OR SOMEPLACE
ELSE?
I MEAN --
>>SO PAUL, IF YOU HAVE -- I
HAVE CONNECTIONS WITH ONE
INVESTORS THAT WANT TO PUT MORE
MONEY --
[LAUGHTER]
>>
>>SORRY TO SAY I DON'T.
>>THIS WAS -- WE CAN TALK LONG,
WE NEED TO TBRAB A BEAR FOR
TH -- NEED TO GRAB ABEER FOR TH.
IT TURNS OUT THE BIOTECH
INVESTMENT FIELD RUN OUT OF
MONEY, PULLED THE MONEY AWAY,
AND THINGS LIKE THIS WERE -- WE
WOULD HAVE TO SPEND DOUBLE THAN
THIS TO BE ABLE TO MAKE IT THEIR
TEU PICK.
I THINK IT'S GOING TO BE THERE
AND I CONVINCED WE'RE GOING TO
TREAT CANCER IN THE FUTURE WITH
AN APPROACH LIKE THIS.
WE JUST NEED TO MAKE SECONDARY
GENE EDITS TO MAKE THOSE T-CELLS
REPEATED ANTIGEN EXPOSURE.
WE CAN DO SYNTHETIC BIOLOGY, WE
CAN PUT RECEPTORS LIKE WHAT
WE'VE DONE WITH CRIS GARCIA
WHERE WE CAN GIVE AN ORTHOGONAL
CYTOKINE THAT TURNS ON THOSE
T-CELLS ONLY SPECIFICALLY.
THE ABILITY TO DO THIS KNOCK-IN
ALLOWS US TO PUT TWO OR THREE
GENES ON THAT PLASMID, AND WE
DON'T LOSE EFFICIENCY.
IT'S ALL THERE TO BE DONE.
SO -- GO BACK TO DOING IT.
>>OKAY, GREAT.
>>THANK YOU.
THERE IS A GOOD BEER PLACE
NEARBY.
>>THANK YOU.
>>SO FROM AN INFORMATION POINT
OF VIEW, I'M TRYING TO
UNDERSTAND THE TWO SYSTEMS OR
THE PATH OF THE SYSTEM THAT YOU
COVERED, THE MUTATIONS ON
NEOANTIGEN AND THE SECOND, THE
TCR.
SO I'M TRYING TO UNDERSTAND
WHICH PART OF THE SYSTEM PLAYS A
MORE STRONGER ROLE THAN THE
OTHER.
SO LET'S SAY YOU HAVE A SINGLE
MUTATION IN A THOUSAND PATIENTS.
HOW MANY PATIENTS WOULD YOU SEE
THAT DO HAVE THE TCR TO TRY TO
TARGET THAT NEOANTIGEN?
>>I DON'T HAVE A WAY TO ANSWER
THAT QUESTION.
MORE IMMUNOGENIC CANCER, MO
TCRs, MORE CLONOTYPES, THAT'S
GOOD AND THOSE PATIENT RESPOND
TO APT PD-1, THEY DON'T NEED
ANYTHING ELSE.
WE HAVE TO TREAT THE ONES WHERE
THIS DOESN'T HAPPEN.
BUT EVEN IN THAT CASE, WHEN WE
HAVE VERY FEW MUTATIONS, VERY
FEW TCRs, I SHOW YOU THOSE
THREE RANDOM PATIENTS WITH
MELANOMA, WE CLONE FIVE TCRs
AND ALL OF THEM KILL THE MATCHED
TUMORS.
SO THERE'S HOPE THERE THAT EVEN
IN THE SITUATIONS WHERE THE
CANCER WASN'T TOO IMMUNOGENIC ON
THE IMMUNE SYSTEM WASN'T DOING
MUCH ABOUT IT, AND A SENSITIVE
APPROACH LIKE THIS THAT CAPTURES
ENOUGH OF THE LANDSCAPE OF THE
MUTATION AND HLA PEPTIDE
DETERMINANTS THAT WE CAN GET
THOSE TCRs AND USE THEM.
>>THANK YOU.
JUST BUILDING ON THAT,
CONSIDERING SUCH A SMALL SUBSET
OF THEM WERE ACTUALLY
IMMUNOGENIC, IT IS A LITTLE
SURPRISING THAT METHODS LIKE TMB
DO HAVE SOME POWER POTENTIALLY,
SO JUST THAT FINAL --
>>MY FRIEND DON SCHUMACHER SAYS
TMB IS LIKE BUYING TICKET TO THE
LOTTERY, THAT YOU HAVE TO BUY
SOME TO BE ABLE TO WIN AND IF
YOU BUY MORE, YOU MAY BE MORE
LUCKY, BUT THAT DOESN'T MEAN
THAT YOU HAVE ONE, YOU CANNOT
WIN IT.
>>THANK YOU.
>>THAT WAS AN AMAZING TOUR
DEFORCE AND THE SCIENCE IS
REALLY FASCINATING.
I HAVE A QUESTION ABOUT YOUR
MANUFACTURING PROCESS.
TWO QUESTIONS.
ONE IS, WHAT MADE IT WORK SO
MUCH BETTER IN VERSION THREE,
AND ALSO WHAT IS THE INFUSION
BACK LOOK LIKE AS FAR AS DATA
DIFFERENTIATION?
>>SO THE MANUFACTURING IMPROVED
WITH CHANGES IN THE MEDIA,
CHANGES IN THE -- AT THE END
THEY HAD THIS BETA TEST OF -- I
FORGOT -- IT'S IN THE PAPER
BUT -- AND THEN OPTIMIZATION OF
EVERY SINGLE STEP.
SO AT SOME POINT, THEY WERE
EXPANDING THE CELLS THAT WERE
TOO CROWDED AND THE DIFFERENT
MANUFACTURING CHANGES WHICH ARE
DIFFICULT TO EVOLVE AT CLINICAL
GRADE AND UNDER THE SAME IND BUT
IT WAS ABLE TO DO THAT, AND AT
THE END, THEY ENDED UP WITH GOOD
CELLS THAT COULD EXPAND IN VIVO.
THE BACK -- INFUSION -- MOSTLY
LIKE THE TCR ENGINEERED T-CELLS
LIKE -- WHERE IT'S NOT LIKE YOUR
TILES THAT WAS A MILKY THING,
CRYOPRESERVED --
>>BUT THERE'S DATA
DIFFERENTIATION IF YOU LOOKED AT
THE MAR MARKERS AND SO FORTH, WE
THEY DIFFERENT THAN A TYPICAL
RETROVIRAL BAG?
>>SO WE PUT ALL OF THE DATA IN
THE PAPER.
IT'S ALL THERE.
IT'S ALL DESCRIPTIVE.
THERE WAS SOMETHING THAT -- I
DON'T KNOW, SOME -- THERE WAS A
DIFFERENT DIFFERENTIATION STAGE.
THERE WAS AN ATTEMPT TO DO STEM
CELL -- THERE WERE QUITE A BIT
OF THEM -- IT DIDN'T
DIFFERENTIATE ANYTHING IN
EXPANSION OR TARGETING --
ACCUMULATION IN TUMOR.
I'M SORRY I HAVEN'T LOOKED AT
THAT DATA FOR SIX MONTHS SO I
CANNOT ANSWER.
>>SO ON SIMILAR LINES OF THE
TUMOR MICRO -- HAVE YOU EVER
THOUGHT ABOUT TESTING THE
METABOLISM OF THESE T-CELLS AT
DIFFERENT STAGES OF YOUR CELL
GROWTH OR MAKING THIS JUST
BEFORE -- WHEN YOU'RE INJECTING
INTO THE PATIENT?
WHAT IS THE METABOLIC STATE OF
THE T-CELLS AT DIFFERENT STAGES?
OF THIS SPECIFIC ONE.
>>SO THESE T-CELLS, I'M
THINKING ABOUT JIM'S QUESTION,
THERE WERE NOT THAT --
PHENOTYPICALLY THEY WERE NOT
THAT DIFFERENT FROM RETROVIRALLY
TRANSDUCED TCR TRANSGENIC
T-CELLS FROM YOUR PAPERS FROM
PAUL ROBYNS AND OTHERS.
ROBBINS AND OTHERS.I DON'T KNOWE
HAD GOOD MANUFACTURING THAT THE
METABOLIC STATE OF THOSE CELLS
WERE ANY ALTERED.
THEY WERE INFUSED IN LOWER
NUMBERS THAT WOULD BE THE
THRESHOLD TO BE ABLE TO HAVE A
CLINICAL RESPONSE.
SOME T-CELL RECEPTORS WERE NOT
TO THE AFFINITY THAT SHOULD BE
AND THEN THE MAJORITY OF
PATIENTS DID NOT GET
SUPPLEMENTAL IL-2.
SO I THINK TO GET TO THE -- WE
FIRST LOOK AT THE CELL THERAPY
RIGHT AND THEN WE CAN GET TO
YOUR QUESTION.
>>OKAY.
THANK YOU.
>>THANK YOU VERY MUCH.
[APPLAUSE]