>>I'M WARREN LEONARD IN
THE NHLBI AND ON
BEHALF OF THE CYTOKINE INTEREST
GROUP I'M DELIGHTED TO WELCOME
TODAY'S SPEAKER CHRIS GARCIA
FROM STANFORD WHO IS A
WORLD-RENOWNED ENGINEER
INTERESTED IN SIGNALING AND
MAKING IMMUNO THERAPEUTICS.
I'M DELIGHTED THAT
WE'RE HOSTING CHRIS GARCIA A
FRIEND WITH WHOM I HAD THE
PLEASURE OF COLLABORATING.
HE IS PROFESSOR OF MOLECULAR AND
CELLULAR PHYSIOLOGY AND YOUNGER
PROFESSOR STRUCTURAL BIOLOGY AT
STANFORD AND INVESTIGATOR TO THE
SHOW ARD HUGHES MEDICAL
INSTITUTE.
HE'S INTERESTED IN CELL SURFACE
RECEPTORS AND THEIR LIGANDS WITH
EXTRAORDINARY WOORK ON CYTOKINES
IN THE T-CELL RECEPTOR AND USING
INNOVATIVE APPROACHES TO
UNDERSTAND HOW CELL SURFACE
RECEPTORS ENGAGE LIGANDS AND
TRANSDUCE SIGNALS.
HE HAS DETERMINED THE STRUCTURE
OF CYTOKINE FAMILY MEMBERS,
COMPLEX RECEPTORS INCLUDING
COMMA GAMMA CHAIN CYTOKINES,
GP10, IL12, AND INTERFERONS.
HE ALSO DETERMINED THE FIRST
RECEPTOR FOR A JACK CYTOKINE
RECEPTOR COMPLEX [INDISCERNIBLE]
STRUCTURE AND MUCH MORE.
HE CREATED IL2 AND IL4
SUPERKINES DEVELOPED A WIDELY
USED ORTHOGONAL IL2 SYSTEM AND
GENERATED A BROAD RAINCHL OF
CYTOKINE AGONISTS AND SERIES
POINTSIGATE AGONIST AS WELL AS A
CIRC ALPHA VARIANT WITH ENHANCED
AFFINITY TO ANTAGONIZE CD47.
HE FOUNDED SYNTHA SIGN A BIOTECH
WITH A NEW FOCUS ON THERAPIES.
CHRIS RECEIVED HIS MD. FROM
TULANE AND Ph.D. FROM JOHNS
HOPKINS.
HE WAS A POST DOC AT GENENTECH
AND THEN AT SCRIPS WITH IAN
WILSON AND ON FACULTY AT SCRIPTS
BEFORE MOVING TO STANFORD WHERE
HE'S BEEN EVER SINCE.
HE'S A MEMBER OF THE NATIONAL
ACADEMY OF SCIENCES, NATIONAL
ACADEMY OF MEDICINE AND RECEIVED
THE 2021 WILLIAM POLE AWARD FROM
THE CYTOKINE AND INTERFERON
SOCIETY.
FIRST SLIDE PLEASE AND ONLY
SLIDE.
FINALLY, I WOULD BE REMISS IF I
DIDN'T MENTION HIS ATHLETIC
PROWESS INCLUDING SWIMMING,
BICYCLE, AND RUNNING WITH MANY,
MANY ULTRA MARATHONS.
SO IT'S A PLEASURE TO WELCOME
YOU, CHRIS, HIS TITLE IS
IMMUNITY THROUGH LIGAND-RECEPTOR
ENGINEERING.
>>THANKS WARREN AND CHRIS FOR
FOR ORGANIZING THIS, IT'S A REAL
HONOR TO GIVE ANOTHER WALS
LECTURE, I GAVE 1 SEVERAL YEARS
AGO AND FRANCIS COLLINS
INTRODUCED ME IN THE BIG
AUDITORIUM AND THIS IS A BIG
THRILL BUT THIS WILL SUFFICE,
TODAY, I HOPE TO BE BACK IN
PERSON SOON.
Y SOPHISTICATED THAT PICTURE
THAT DOES BRING BACK MEMORIES
THAT WAS THE CASCADE CREST 100
MILLER IN THE CASCADE MOUNTAINS
EVEN THOUGH I LOOK VERY
COMFORTABLE, I WAS VERY
UNCOMFORTABLE AT THE TIME IT WAS
TAKEN.
SO WHY DON'T WE GET ON WITH THE
TALK HERE
JUST TO GET A BIT OF A
BACKGROUND, I THINK MY INTEREST
CAN BE SUMMARIZED AS THINGS THAT
HAPPEN AT THE CELL FUR FASMOSTLY
LIGAND REMEDIATED RECEPTOR
EVENTS AND HOW PROTEINS ENGAGE
RECEPTORS AND HOW THEY
REACTIVATE SIGNALING AND USING
THAT INFORMATION AND USING THE
ACTIONABLE VALUE FROM THAT
INFORMATION TO TRY AND PERTURB
THE PHYSIOLOGY OF THE CELL.
AND THAT'S KIND OF WHAT WE'RE
INTO HERE, ON MANY DIFFERENT
LEVELS AND SYSTEMS.
LET ME GET MY LASER POINTER
HERE.
AN INTERPLAY OF STRUCTURAL
BIOLOGY WITH PROTEIN ENGINEERING
TO UNDERSTAND MECHANISM AND THEN
USE THE TERM ACTIONABLE VALUE
AND THATIA REALLY MY INTEREST IN
MECHANISM IS EXTRACTING
INFORMATION THAT WE CAN USE FOR
TRANSLATION TO POSSIBLY MAKE
DRUGS.
AND THE SYSTEMS IN THE LAB TO
KIND OF NAME SORT OF THE
HIGHLIGHTS HERE, I'VE WORKED IN
IMMUNITY INFECTION FOR A LONG
TIME, REALLY GO WAY BACK WITH
THE T-CELL RECEPTOR AND THEN,
YOU KNOW THROUGH MY EXPERIENCE
AS A GENENTECH, I DEVELOPED AN
INTEREST IN CYTOKINE SIGNALING.
SO THOSE ARE THE TWIN POLES AT
THE LAB BUT WE ALSO HAVE A BIG
PROGRAM IN REGENERATIVE MEDICINE
LOOKING AT THE NOTCH PATHWAYS
AND THEN ALSO, MOST OF THE
RECEPTOR SYSTEMS IN THE LAB ARE
ACTIVATED BY SOME TYPE OF
INDUCED PROXIMITY SO WE'RE
REALLY DIGGING INTO THAT NOW AS
AN ENGINEERING PLATFORM TO
INDUCE PROXIMITY BETWEEN
PROTEINS ON CELLS THAT DON'T
NORMALLY COME TOGETHER AND I'LL
TALK SOME ABOUT THAT TODAY.
SO OF ALL THESE DIFFERENT KIND
OF PROJECTS IN THE LAB, I WILL
MAINLY FOCUS ON CYTOKINES TODAY
AND REALLY ADDRESS SOMETHING
THAT I REALLY GOD INTERESTED IN
READING WARREN LEONARD'S PAPERS
AND USING THIS WORD PLEOTROPY,
HE WAS 1 OF THE FIRST PEOPLE TO
PUT THIS WORD OUT THERE AND IT'S
AN APT WORD BECAUSE CYTOKINES
ARE PLEOTROAPIC IN THAT THEY ACT
ON MULTIPLE DIFFERENT CELL
TYPES.
CELLS HAVE COUNTER VAILING
ACTIONS, THEY HAVE SPECIFIC
ACTIONS AND IT'S A VERY
DIFFICULT ISSUE IN CYTOKINE
BIOLOGY TO UNDERSTAND WHAT THE
SPECIFIC FUNCTIONS OF DIFFERENT
CYTOKINES ARE IN THE FACE OF
THIS PLEIOTROPY.
BUT IT'S ALSO BEEN A HUGE
BARRIER TO GETTING CYTOKINES IN
THE CLINIC OTHER THAN OTHER
PARTICULAR EXAMPLES SUCH AS EFO
AND A FEW OTHERS THAT HAVE VERY
RESTRICTED EXPRESSION OF THEIR
RECEPTORS.
IMMUNE CYTOKINES LIKE IL2 AND
OTHERS HAVE BEEN VERY DIFFICULT
TO IMPLEMENT IN THE CLINIC IN AN
AGONIST MOST AND THAT'S
SOMETHING I'VE BEEN VERY
INTERESTED IN BASED ON THE
MECHANISM OF HOW CYTOKINE
SIGNAL.
SO TO THE RIGHT HERE WHAT I'M
SHOWING YOU HERE IS THE KIND OF
CLASSIC PARADIGM OF HOW CYTOKINE
SIGNAL, BINDS WITH HIGH AFFINITY
TO 1 RECEPTOR WHICH RECRUITS IN
THE SECOND RECEPTOR THAT CAUSES
THE JANUS KINASES WHICH ARE
ASSOCIATED WITH THE
INTERRACELLULAR DOMAINS TO
PHOSPHORYLATE AND THEN THE STATS
WITH THAT THEY GO INTO THE
NUCLEUS.
AND THIS 2 STEP MECHANISM
AFFORDS A LOT OF ENGINEERING
PLEIOTROPY AGONIST TO BRENG
THESE 2 TOGETHER IN DIFFERENT
WAYS THAT COULD MODULATE
SIGNALING.
AND WE'VE RELIED A LOT ON
STRUCTURAL BIOLOGY TO PROVIDE
BLUEPRINTS OF CYTOKINE RECEPTOR
COMPLEXES, THIS A SELECTION, AN
INCOMPLETE SELECTION FROM OVER
THE YEARS STARTING WITH IL1 AND
YOU'LL SEE PROBABLY SOME OF
YOUR--1 OF YOUR FAVORITE
MOLECULES UP HERE, WE PROBABLY
LOOKED AT IT AND YOU KNOW WE
REALLY TRY TO INNOVATE FROM THE
STRUCTURES OF THE EXTRA CELLULAR
COMPLEXES AND THEN
RECENTLY--SORRY, I THINK I WILL
KEEP MY POINTER--WELL, HOW DO I
GET THIS THING GOING HERE.
PLAYER OPTIONS, LET'S GO OFF
THERE, THERE WE GO.
OKAY.
SO WE RECENTLY LAST YEAR A GROUP
IN MY LAB, CALEB,
[INDISCERNIBLE] AND OTHERS NOW
NATHANIEL DAB NET WE DETERMINED
THE FULL LENGTH KINASE OF A
CYTOKINE RECEPTOR AND I WILL
TALK MORE ABOUT THAT LATER BUT
THE REASON I SHOWED THIS NOW IS
TO COMPARE AND CONTRAST HOW WE
THINKING ABOUT CYTOKINE
ENGINEERING WHICH IS IN THE
CONTEXT OF NOW GPC Rs ARE
ACTIVATED AND THAT THEY'RE VERY
PLASTIC MOLECULES AND YOU KNOW
KIND OF THE WHOLE FOUNDATION OF
THE GPCR DRUG DISCOVERY INDUSTRY
IS THIS IDEA THAT YOU CAN MAKE
BIASED AGONISTS AND PARTIAL
AGONISTS, THIS IS A CLASSICAL
PHARMAICOLOGICAL DOSE RESPONSE
GROUP FOR GPCR LIGANDS THAT HAS
REALLY BEEN LIMITED TO G-PROTEIN
RECEPTORS AND ION CHANNELS AND
WE'RE THINKING ABOUT THIS KIND
OF ENGINE, THIS SIGNALING ENGINE
IN THE CONTEXT OF THESE
PHARMAICOLOGICAL PRINCIPLES AND
THAT'S REALLY DRIVEN A LOT WITH
WHAT I WILL SHOW YOU TODAY, SORT
OF ENGINEERING THE PROTEIN TO
ACHIEVE THE SAME TYPE OF
BEHAVIORS THAT SMALL MOLECULE
AGONIST FOR GPC Rs WORK.
AND ALSO NOT JUST ENGINEERING
THE CYTOKINE BUT ALSO MAKING
MOLECULES THAT CAN CHANGE THE
GEOMETRY OF THE CYTOKINE
RECEPTOR DIMER.
SO I WILL JUST REALLY GET
STARTED HERE WITH A FEW EXAMPLES
OF HOW WE'RE DOING THIS AND
STARTING OFF ON IL2 AND WE
DETERMINE THE STRUCTURE OF THIS
HIGH AFFINITY COMPLEX QUITE A
FEW YEARS AGO NOW BUT IT'S JUST
CONTINUED TO REALLY YIELD
INFORMATION THAT HAS ALLOWED US
TO ENGINEER A WHOLE SLEW OF
INTERESTING IL2 VARIANTS EXPW I
WON'T SUMMARIZE A LOT OF THAT
JUST FOCUSING ON THE MORE RECENT
DATA AND REALLY THE INTEREST HAS
BEEN TO ASK IF WE CAN MAKE IL2
VARIANTS THAT MIGHT SKEW T-CELLS
TOWARDS REGULATORY T-CELLS OR
EFFECTOR T-CELLS AND SO HAVE A
LOT OF OTHER GROUPS AND THAT'S
TO MAKE DRUGS FOR AUTOIMMUNITY
OR CANCER IMMUNOTHERAPY AND 1
RECENT SERIES OF STUDIES WE'VE
DONE IS WE'VE MADE A SERIES OF
PARTIAL STAT AGONIST BY MUTATING
THE COMMON GAMMA CHAIN BINDING
FACE OF IL2 WHERE WE ESSENTIALLY
BEEN ABLE TO TITRATE STATED 5 IN
INCREMENTS AND WE SYSTEMATICALLY
CHARACTERIZED THESE INVITRO BUT
ALSO ADMINISTER THEM TO MICE TO
OBSERVE THE IMMUNE CELL
PROFILING BEHAVIOR AND HAD SOME
INTERESTING RESULTS HERE.
FOR INSTANCE, THIS 1 MOLECULE
CALLED HLINT, WHICH WE
COLLABORATED WITH ON THIS, IT'S
A VERY SMALL, A PARTIAL AGONIST
THAT'S BARELY, VERY SLIGHTLY
ATTENUATE IN THE STAT 5 BUT YET
WARREN TOOK THIS MOLECULE AND HE
AND HIS GROUP FOUND SOME VERY
INTERESTING PROPERTIES THAT IT'S
VERY IL2 LIKE IN ITS ABILITY TO
EXPAND T-CELLS BUT THERE'S A
REDUCTION AND ACTIVATION OF
EXHAUSTIAN MARKERS LIKE TIM 3
AND LAG, AND IT ALSO HAS THESE
STEM LIKE PROCESSES LIKE THE
TRANSCRIPTION FACTOR AND
RESPONSE WHICH IS REDUCED DPLI
COLSIS WHICH IS HALLMARKS OF
STEMNESS, AND IN WORKING WITH
NICK RESTIFO, THEY FOUND THAT
PRECONDITIONING CELLS PRIOR TO
ADOPTED TRANSFER IN A FEMALE
MODEL SIGNIFICANTLY DELAYED
TUMOR GROWTH COMPARED TO IL2 OR
SUPER 2 ASK ALSO LED TO ENHANCED
PERSISTENCE.
SO THAT WAS REALLY JUST THE
SLIGHTEST ATTENUATION OF STAT 5
SIGNALING REALLY SIGNIFICANTLY
CHANGEDLET IN VIVO ACTIVITY OF
THIS MOLECULE WHICH REALLY SHOWS
HOW TUNEABLE THESE CYTOKINE ARE.
HERE'S ANOTHER EXAMPLE THAT'S A
BIT MORE ATTENUATE IN STAT 5
WHICH WE CALL REH AND CALEB
GLASSMAN IN MY LAB AND WE WORK
WIDE WARREN ON THIS, FOUND IT
HAS HIGH SELECTIVITY FOR
REGULATORY T-CELLS VERSUS
CONVENTIONAL T-CELLS AND WE DID
SOME RNA SEQ WITH WARREN AND
FOUND THAT IN T-CONS WE SAW A
REDUCTION IN THE TRANSCRIPTION
OF CERTAINLY THESE
PROINFLAMMATORY MOLECULES
WHEREAS THAT WAS LESS SO IN THE
REGULATORY T-CELLS SO HERE
AGAIN, THIS SLIGHT ATTENUATION
OF THE MEMBRANE PROAKS
MALSIGNALING ADAPTOR REALLY LED
TO THESE DIFFERENT CHANGES IN
DIFFERENT CELL SUBSETS OF
T-CELLS AND WE ALSO SHOWED THAT
THERE WAS THIS MOLECULE SHOWED
IMPROVED TREATMENT OF A
DSS INTESTINAL INFLAMMATION
COMPARED TO IL2 AND THAT IS MOST
LIKELY DUE TO THE T-REG
EXPANSION BUT WE ALSO HAD AN
INTERESTING EXPERIENCE WITH THIS
MOLECULE THAT, YOU KNOW IT
LOOKED INITIALLY TO US LIKE A
TREG POTENTIATOR BUT SO THIS AN
MP38 TUMOR MOLLEDLE--IT EXPANDS
THEM IN THE TUMOR AND LESS TOXIN
THAT WE'VE LOOK TD AT SO THIS
WAS QUITE A SURPRISE AND COUNTER
ENTUMOR SPECTRUMMATIVE AND
REALLY KIND OF TURNS THIS IDEA
IN THE FIELD THAT IT'S REFERRED
TO AS THE ALPHA PROBLEM BECAUSE
IT'S HIGH ON TREGS WELL HERE WE
HAVE A CD25 VERSION OF IL2 THAT
EXHIBITS VERY PROFOUND ANTITUMOR
EFFECTS AND LESS TOXICITY.
SO ALPHA MAY NOT BE THE PROBLEM,
IT MAY ACTUALLY BE AN
OPPORTUNITY TO SELECTIVELY
TARGET THE DETAILS IN THE
ACTIVATED TAILS IN THE TUMOR.
NOW MOVING ON TO ANOTHER
CYTOKINE HERE, IL12, IT'S
ANOTHER VERY POTENT T-CELL
CYTOKINE THAT HAS REALLY BEEN A
TANTALIZING THERAPEUTIC TARGET
EXCEPT IT'S EXTREMELY TOXIC EVEN
MORE SO THAN IL12.
CALEB GLASSMAN IN MY LAB, A
STUDENT HERE DETERMINED THE
STRUCTURE OF THE IL12
[INDISCERNIBLE] COMPLEX BY
CRYOEM AND I WILL NOT DWELL ON
THE STRUCTURAL DETAILS HERE
OTHER THAN TO SHOW YOU THAT THE
INTERACTION OF THE IL12 R-BETA 1
WITH THE CYTOKINE IS RATHER
UNUSUAL IN THAT SO THE GREEN
MOLECULE HERE IS IL12 RBETTA 1
AND P40 IS THE RECEPTOR SUBUNIT
OF THE CYTOKINE AND IT'S
INTERACTING WITH THE IG-DOMAINS
OF THE CYTOKINE RATHER THAN THE
HELICAL BUNDLE LIKE MOST
CYTOKINE DO.
SO THAT LED TO AN INTERESTING
OPPORTUNITY OPPORTUNITY HERE TO
ASK IF WE COULD DECOUPLE THE
ACTIVITY OF IL12 FROM NK CELLS
WHICH SECRETE A LOT OF GAMMA
INTERFERON WHICH IS MOST LIKELY
DUE TO THE--MOST LIKELY LEADS TO
THE TOXICITY OF IL12, AND
SELECTIVELY ACTIVATE T-CELLS.
SO, WHAT CALEB DID IS HE DID
STAINING OF ACTIVATED T-CELLS
VERSUS NK CELLS AND FOUND THAT
IL12 R-BETA 1 IS VERY HIGH ON
CD8 T-CELL BLASTS, MUCH LESS 1
ON NK CELLS SO WHAT ABOUT WITH
WITH THE STRATEGY, IF WE COULD
WEAKEN THE INTERACTION OF IL12
WITH OUR BETA 1, WE MIGHT GET
SELECTIVITY FOR THE CD8 T-CELLS
SO HE MADE A SERIES OF PARTIAL
AGONISTS BY MUTATING THE P40
SUBUNIT OF IL12, NOT THE HELICAL
SUBUNIT AND YOU CAN SEE THIS 2
XL OR 3 XL OR 4 XL, NOW STAT 4
PARTIAL AGONISTS.
AND WHEN CALEB LOOKED AT A MOUSE
AT THEIR ABILITY TO ACTIVATE
T-CELLS VERSUS NK CELLS YOU SEE
THEY LARGELY RETAIN THE ABILITY
ON T-CELLS TO MAKE GAMMA
INTERFERON BUT THE NK-CELLS
THERE'S MUCH LESS ACTIVITY TO
THE NK-CELLS SO WE TOOK THIS
ENDOTUMOR MODEL AND THIS MC 38
TUMOR MODEL AGAIN AND FOUND THAT
WHILE WILD-TYPE IL2 IS EXTREMELY
TOXIC, EACH AT 1 MICROGRAM, MICE
GET VERY SICK UNTIL THEY
RECOVER, AND THEN AT HIGHER
DOSES, IT'S LETHAL, OUR PARTIAL
AGONIST HAD NO TOXICITY
WHATSOEVER.
BUT THEY MAINTAINED THE SAME
ABILITY TO REDUCE TUMOR GROWTH
AND ENHANCE SURVIVAL.
SO ESSENTIALLY WE DECOUPLED THE
TOXICITY OF IL12 FROM THE
ANTITUMOR EFFECTS AND JUST IN
FULL DISCLOSURE, I STARTED A
COMPANY CALLED SYNTHYKINE TO
TEST THIS IN HUMANS.
NOW LET'S SAY FROM A CANCER
IMMUNO THERAPEUTIC CYTOKINES AND
IMMUNOSUPPRESSIVE, IL10, THIS IS
A NICE STORY DONE BY BOBBY
SAXTON IN MY LAB WHO IS NOW AN
ASSIST ABT PROFESSOR AT UC
BERKELEY SO IL10 IS A WELL KNOWN
IMMUNOSUPPRESSIVE ESPECIALLY ON
MONOCYTES, MACROPHAGES BUT IT
ALSO DOES HAVE SOME ACTIVATING
EFFECTS ON T-CELLS AND THAT'S
LARGELY DUE TO THESE ACTIVITIES
ARE LARGELY CAN BE SEPARATED,
THE STAT 3 IS THE
ANTIINFLAMMATORY EFFECT, DOWN
STREAM OF IL10 RECEPTOR AND STAT
1 IS THE PROINFLAMMATORY SO
BOBBY TOOK A SIMILAR STRATEGY ON
IL10 THAT I'VE SHOWN YOU
ALREADY, HE DETERMINED THE
CRYOEM STRUCTURE OF THE IL10
RECEPTOR COMPLEX, AND IT'S THIS
INTERESTING DISCIPLINARY MERRIC
STRUCTURE SO INSTEAD OF 2
RECEPTORS AND 1 CYTOKINE, YOU'VE
GOT A 2-2-2.
SO IT SORT OF DEPARTS FROM THE
IL2 TYPE OF MODEL I SHOWED YOU
EARLIER BUT WHAT BOBBY DID WAS
HE ALSO--HE FOUND ON MONOCYTES
THAT IL10 RBETTA IS HIGH AND SO
WHAT WE DID WAS HE CREATED A
SERIES OF PARTIAL AGONISTS ON
IL10 TO WEAKEN THE INTERACTION
WITH IL10 R BETA AND THEN FOUND
WHEN HE LOOKED AT DIFFERENT CELL
TYPES THAT HE WAS ABLE TO
PRESERVE STAT 3 INDUCTION ON
MONOCYTES, BUT SIGNIFICANTLY
DIMINISH IT ON T-CELLS, B-CELLS
AND NK-CELLS AND ESSENTIALLY
ERASE STAT 1 SIGNALING FROM
T-CELLS.
SO THAT PREDICTS WE WOULD HAVE
PRESERVED THE ANTIINFLAMMATORY
EFFECTS AND ELIMINATED THE
PROINFLAMMATORY EFFECTS AND THAT
WAS WORN OUT BY THE GENE
SIGNATURES HERE WHEN WE DID IN
RNA SEQ, THESE PROINFLAMMATORY
CYTOKINES WERE REALLY
SIGNIFICANTLY REDUCED WITH A
PARTIAL AGONIST, AND WE ALSO
LOOKEDDA THE ACTUAL CYTOKINES
THEMSELVES AND YOU CAN SEE
COMPARED TO WILD-TYPE, THE
PARTIAL AGONIST OR SIGNIFICANTLY
REDUCED BUT IN A MODEL AT THIS
LPS ACTIVATION MODEL, MONOCYTES
WHERE THEY DUMP OUT TNF AND IL6
AND OTHER PROINFLAMMATORY
CYTOKINES, THE PARTIAL AGONIST
MAINTAINS ITS ACTIVITY AND
INHIBITING THE SECRETION OF
THESE CYTOKINES JUST AS WELL AS
IL10.
AND HE ALSO WENT ON AND DID A
MOUSE SEPSIS MODEL WHERE HE
SHOWED WE WERE ABLE TO PROTECT
THESE MICE FROM THE IL6 AND TNF
CYTOKINE STORM WHEN THE MICE ARE
INFECTED WITH SEPSIS AS WELL AS
WILD-TYPE 10.
SO HERE AGAIN WE'VE SHOWN WE CAN
DECOUPLE THE ACTIVITIES OF THE
CYTOKINES IN GETTING BACK TO THE
NOTION OF PLEIOTROPY THAT I
TALKED ABOUT IN THE BEGINNING,
IT SEEMS THAT CYTOKINE ENGINEER
SUGGEST AN EFFECTIVE WAY OF
ADDRESSING THAT PROBLEM AND
HERE'S 1 LAST PIECE OF DATA ON
IL10 IN A COLITIS MODEL AT IL10S
HAVE A LOT OF INTEREST IN THE
CLINIC ARE INFLAMMATORY BOWEL
DISEASE BUT THERE'S BEEN IN
PROBLEM WITH THIS
PROINFLAMMATORY ASSPECS OF
WILD-TYPE IL10 AND HERE WE
SHOWED THAT WE CAN PROTECT MICE
FROM DSS AS WELL AS CYTOKINE
TYPE HERE AND THIS IS ALSO ON A
CLINICAL PATH THROUGH
SYNTHECOIN, AND FINALLY I WANT
TO FINISH UP ON
INTERFERON-GAMMA, AND WHICH IS
REALLY THE PREMIER MASTER
REGULATOR AND MORE PLEIOTROPIC
THAN IL2 AND IT ACTIVATES ON
MOST TYPES OF IMMUNE SELLS, SO,
WHEN WE STARTED THERE WAS NOT A
STRUCTURURE KNOWN OF THIS
RECEPTOR COMPLEX AND JUAN MEN
DOZA IN MY LAB WHO'S A JUNIOR
FACULTY AT UNIVERSITY OF CHICAGO
TOOK ON THIS CHALLENGE AND
REALLY IT'S SORT OF A BIG VISION
WE HAD FOR THIS, THE RATIONAL
FOR GETTING INTO THIS WOULD BE
TO ASK, COULD WE UNCOUPLE THE
IMMUNOSTIMULATORY FROM THE
IMMUNEUE O SUPPRESS ANT ACTIONS
OF GAMMA INTERFERON, I MEAN THE
PRINCIPAL ACTION OF GAMMA
INTERFERON WHICH MOST PEOPLE
KNOW ABOUT IS THAT IT
UPREGULATES CLASS 1 EXPRESSION,
BUT IT ALSO UPREGULATES CHECK
POINT LIGANDS, SO IT'S
IMMUNOSUPPRESSIVE IN THE TUMOR
MICROENVIRONMENT.
SO WE WANTED TO ASK BY STRUCTURE
IF WE COULD POSSIBLY DECOUPLE
THESE ACTIONS.
SO 1 DETERMINE THE STRUCTURE OF
THE GAMMA INTERFERON RECEPTOR
COMPLEX AND YOU CAN SEE HERE
GAMMA INTERFERON BEAR
RESEMBLANCE TO IL10, IT'S A
TIMER AND DECORATED BY THE 4
RECEPTORS, 2 GAMMA R1S, 2 GAMMA
R2s AND I WILL NOT GET INTO
THE DETAILS OF THIS 1 EITHER BUT
JUST TO SHOW YOU WHAT UPON JUAN
DID HE, HE SELECTIVELY APLATED
THE DIFFERENT BINDING SITES LIKE
GAMMA INTERFERON AND IN SO DOING
CREATED A SERIES OF PARTIAL
AGONISTS, THIS IS PSTAT 1 WE'RE
LOOKING AT HERE.
THIS WAS A BEAUTIFUL CALIBRATION
OF THE MAX RANGE OF GAMMA
INTERFERON.
AND SO, SO WITH THESE PARTIAL
AGONIST THEN, WE WENT AND DID IN
GENE--RNA SEEK EXPERIMENTS TO
LOOK AT GENE EXPRESSION AND
FOUND SOMETHING RATHER
UNEXPECTED WHICH IS THAT MOST OF
THE GENES UPREGULATED BY GAMMA
INTERFERON WERE UPREGULATED TO
ROUGHLY THE SAME EXTENT BY
WILD-TYPE GAMMA VERSUS THE
PARTIAL AGONIST.
WE CALL THOSE STABLE GENES BUT
THERE WAS A MUCH SMALLER STABLE
SUBSET WHERE THE PARTIAL
AGONISTS UPREGULATED GENES TO A
LOWER DEGREE THAN THE WORLD TYPE
AND AMONGST THE STABLE 1S WAS
CLASS 1 MHC, AND AMONGST THE
TUNEABLE 1S WAS PDL 1, THE CHECK
POINT LIGAND.
SO WITH THAT INFORMATION, JUAN
AND [INDISCERNIBLE] IN MY LAB
AND COLLABORATING WITH
[INDISCERNIBLE] STARTED STAINING
CANCER CELL LINES.
HERE'S AN EXAMPLE OF LUNG CANCER
CELL LYNN WHERE THE UPREGULATION
OF HRA 1 CLASS 1 IS EQUIVALENT
BETWEEN WILD-TYPE AND THE
PARTIAL AGONIST WHERE THE PDL1
IS ATTENUATE SIGNIFICANTLY BY
THE PARTIAL AGONISTS.
AND JUAN WENT ON TO STAIN AND DO
THIS EXPERIMENT WITH MANY
DIFFERENT TYPES OF CANCER CELL
LINES AND THERE'S BIAS TO
DIFFERENT DEGREES, SOME MORE,
SOME LESS, SOME NONEXISTENT BUT
WE DO SEEM TO BE ABLE TO TILT
THE BALANCE TOWARDS CLASS 1 OVER
PDL 1 GENERALLY AND SO, WHAT
WE'RE DOING NOW IN MY LAB,
[INDISCERNIBLE] IN MY LAB NOW
MADE SOME MOUSE GAMMA INTERFERON
PARTIAL AGONIST TO ALLOW US TO
PROPERLY EXPLORE THE PROPERTIES
OF THESE BIAS IN TUMOR MODELS IN
IN VIVO.
SO JUST TO SUMMARIZE WHAT I
SHOWED YOU, WHAT HAVE WE
LEARNED, CELLS HAVE INTRINSIC
RESPONSE THRESHOLDS TO CYTOKINES
THAT CAN BE--IT'S A MULTILEVEL
CONTROL OF THESE HARD WIRED
RESPONSE THRESHOLDS BUT THE
BOTTOM LINE IS THAT BUT BY
MAKING PARTIAL AGONIST AND
CYTOKINE WE CAN IN MANY CASES
PHASE OUT ACTIONS ON DIFFERENT
CELL TYPES AND SIGNIFICANTLY
NARROW THE PLEIOTROPY IN THIS
CASE FOR EXAMPLE, BOBBY SAXON
WAS ABLE TO NARROW THE ACTIVITY
OF IL10 TO ESSENTIALLY ONLY
MONOCYTES AND SO REALLY I THINK
THE PATH FORWARD FOR GETTING
CYTOKINES INTO THE CLINIC IS
GOING TO INCORPORATE THIS
CONCEPT HERE OF TUNING DOWN THE
AGANISM AND GETTING BETTER CELL
SET SELECTIVITY.
SO UNTIL NOW WHAT I'VE SHOWN YOU
IS ENGINEERING THE CYTOKINE
ITSELF?
SORT OF TUNING IT DOWN, LOWERING
ACTIVITY, BUT THERE ARE A LOT I
WAS TO DO THIS AND IF YOU GET
ASHES WAY FROM THE CYTOKINE, AND
THE CYTOKINE RIGHT HERE TO THE
RIGHT HERE'S A 4 HELIX CLASSICAL
SYSTEM, TO THE 2 RECEPTORS, AND
ALSO HERE WE NOW HAVE A GBCR AND
I'M SHOWING YOU THE
CONFIRMATIONAL CHANGES THAT GPC
Rs UNDERGO WITH THE G-PROTEIN
IN RESPONSE TO DIFFERENT
LIGANDS.
AND WE TOOK ANOTHER APPROACH
HERE WHICH IS TO ASK INSTEAD OF
JUST DOWN TUNING THE BINDING OF
THE SECOND RECEPTOR, COULD WE
REORIENT THE DIMER
DEO--METABOLIZEDETRY TO
ACCOMPLISH SOMETHING ANALOGOUS
TO THE SHAPE CHANGES THE GPC
Rs UNDERGO WHEN THEY BIND
THEIR LIGANDS.
BUT TO DO THAT, THE SCAFFOLD
ITSELF WAS NOT SUITABLE.
THERE WAS NOT A LOT OF
OPPORTUNITIES TO REORIENT THE
DIMER GERONTOLOGYSTS OHMETRY
WITH THE SCAFFOLD.
SO WHAT WE DID, SO WHAT WE WOULD
LIKE TO DO AND WE WOULD LIKE TO
HAVE A SCAFFOLD THAT WE COULD
CHANGE THE DIMER GEOMETRY AND
CHANGE IT TO THE JACK TO 1
ANOTHER TO TUNE THE SIGNALING UP
IN THE WAY THE WAY THE CHEMISTRY
IS ABLE TO MAKE SMALL MOLECULES
THAT CAN CHANGE THE SHAPE OF GPC
Rs, SO THAT WAS THE OVERALL
CONCEPT AND WE WENT ABOUT THIS,
BY USING A DIFFERENT BINDING
SCAFFOLD WHICH IS CALLED A
DARKEN AND THE DARKEN IS A
PROFESSIONAL BINDING SCAFFOLD,
THAT WAS CREATED FOR THE SOLE
PURPOSE OF SELECTING SCAFFOLDS
FROM LIBRARIES.
SO [INDISCERNIBLE] IN MY LAB
MADE A DARPIN LIBRARY AND MADE A
CRISTTAL STRUCTURE OF THE
COMPLEX WITH EPO RECEPTOR.
AND THEN WE COLLABORATED WITH
DAVID BAKER'S LAB TO SEE IF WE
COULD DIMERIZE THE DARPINS AND
BRING IN A SECOND RECEPTOR TO
MAYBE ACTIVATE SIGNALING AND WE
ARE ABLE TO DO THAT.
WE DESIGNED WITH THE BAKER LAB,
THESE MUTATIONS IN THE DIMER
INTERFACE THAT COMPELLED THE
DIMERIZATION OF THE DARPIN AND
THAT BROUGHT 2 RECEPTORS
TOGETHER AND THAT HAS BEEN A
FANTASTIC YOU CAN CHANGE THEM
AND WE DID SO AND YOU CAN SEE
HOW THEY MEASURE THE STAT 5 OF
THE ERK SIGNALING, AS WE
INCREASE THE DIMER ANGLE WE
PHASE OUT SIGNALING SO AS THE
RECEPTORS START TO LIE FLATOT
EMPLOYMENT PROGRAMS BRAIN, THE
JACKS SEEM TO BE UNABLE TO
COMMUNICATE WITH EACH OTHER
ANYMORE AND WE DON'T GET
ACTIVATION OF THE RECEPTOR.
AND THEN ANOTHER--THERE'S
ANOTHER WAY OF INSERTING THESE
INTO THE DARPINS THAT GIVES YOU
A CHANGE IN THE DISTANCE BETWEEN
THE MONITOR MERS RATHER THAN THE
CURVEATURE, SO THAT ALLOWS THE
PURE SEPARATION FORT RECEPTORS
WHICH YOU CAN MEASURE AND IN
THAT SERIES, RATHER THAN A IMRAD
Y'ALL PHASING DOWN OF THE EMAX
WE GOT PRETTY MUCH MAINTAINED
WITH INCREASING DISTANCE, THE
EMAX OF SIGNALING THROUGH STAT 5
AND THEN AT SOME THRESHOLD
DISTANCE, IT BROKE OFF AND
CLEARLY THE JAX AT THAT POINT
WERE NOT ABLE TO TALK TO EACH
OTHER SO WE HAVE 2 DIFFERENT
KIND OF RECEPTOR TUNING THROUGH
THIS IS A GERONTOLOGYSTS
O--METABOLIZEDETRY AND THIS IS
DISTANCE.
AND WE WENT ON TO ASK ABOUT THE
PHYSIOLOGICAL RELEVANCE OF THIS
KIND OF APPROACH WORKING
[INDISCERNIBLE] AT THE BROAD, WE
HAD THIS ASSAY FROM
HEMATOPOIETIC STEM CELLS TO
RECELLS WHERE YOU FOLLOW CD31
AND 235 AND THEY MAKE THIS
STEREO TYPICAL U-SHAPED CURVE
OVER THE COURSE OF THE
DEVELOPMENT AND THIS IS SHOWING
YOU WHAT EPO LOOKS LIKE AFTER
THE 14th DAY OR SO.
WELL WE FOUND BY USING THESE
DARPINSINS, WE USE THE RECEPTOR
IN DIFFERENT INCASULAR GEOMETRYS
WE CAN ABORD RED BLOOD CELL
DEVELOPMENT AT DIFFERENT STAGES
FROM ESSENTIALLY EARLY STAGE TO
UPRIGHT ANGLES WHICH COPIES EFO,
SO THIS IS AN EXAMPLE WHERE
CHANGING THE ORIENTATION REALLY
HAD PROFOUND EFFECTS ON CELL
FATE DETEARM NATIONS AND THAT'S
SOMETHING THAT I REMAIN VERY
INTERESTED IN.
BUT IN DOING IT IN A SLIGHTLY
DIFFERENT WAY.
SO THE DARPINSINS ARE
INTERESTING BUT THEY DON'T HAVE
DRUG LIKE PROPERTIES, THEY ARE
KIND OF A SYNTHETIC TOOL
COMPOUND WHICH YOU CAN DO VERY
INTERESTING PROTEIN ENGINEERING
THINGS WITH THEM BUT THE STEP
TOWARDS PUTTING THISEM INTO
HUMANS IN SYSTEMIC CIRCULATION
IS REALLY NOT BEEN PROVE KNOW
YET THAT'S POSSIBLE SO WE WANT
TO ACHIEVE WHAT I JUST SHOWED
YOU WHICH IS THIS GEOMETRIC
REORIENTATION OF THE EXTRA
CELLULAR DOMAINS BUT NOT USING A
DARPINS SCAFFOLD SO FOR THIS WE
TURN TO A NANO BODY DIMER
SCAFFOLD, EITHER 2 NANO BODIES
TOGETHER AS A STRING LIKE A NANO
BODY OR SEFB, ESSENTIALLY SMALL
BINDING DOMAINS THAT WILL
HOPEFULLY DIMERIZE 2 RECEPTORS
HERE LIKE IN THIS CASE IL2
BRINGS TOGETHER IT'S 2
RECEPTORS.
AND SO THAT WHEN WE LINK THEM
TOGETHER THEY WILL BRING THE 2
RECEPTORS TOGETHER IN DIFFERENT
GEOMETRYS BY VIRTUE OF THE FACT
THAT THEY'RE BINDING TO
DIFFERENT PARTS OF THE EXTRA
CELLULAR DOMAIN.
IS IT WE CAN MAKE A MATRIX THEN
OF THE NANO BODIES, THE BETA
BINDERS VERSUS THE GAMMA BINDERS
IN 2 DIFFERENT ORIENTATIONS, A
FORWARD AND REVERSE ORIENTATION
AND IN THAT WAY WE GET THIS
MATRIX OF GEOMETRYS THAT WE CAN
VERY EASILY TEST ON MOLECULES ON
CELLS FOR STAFF 5 ACTIVATION AND
WHEN MICHELLE DID THIS SHE FOUND
A REAL DIVERSITY OF ACTIVATION
STRENGTHS OF BINDERS THAT HAVE
THE SAME MAX AS IL2 TO SOME OF
THEM SUPER AGONIST THAT WERE
STRONGER THAN IL2 AND THIS
WASN'T JUST AN ACTIVATION
STRENGTH ISSUE.
WE SEE BIAS, TOO, SO WHEN
MICHELLE LOOKED AT FOSTER NURSED
FOCUSED ON STAT 5 AND FOSTER
NURSED FOCUSED ON AKT, CAN YOU
SEE THAT AS WE LOOK AT DIFFERENT
VERSIONS OF THESE MOLECULES, WE
CAN SEE DIFFERENT DEGREES OF
BIAS OF STAT 5 OVER
IEE, AUDIENCE KT.
AND WHEN MICHELLE LOOKED AT THE
FUNCTION, DID SOME FUNCTIONAL
ASSAYS WITH THESE MOLECULES FOR
EXAMPLE, LOOKING AT CD8 T-CELL
DIFFERENTIATION FROM PBMCs,
SHE FOUND THAT DIFFERENT
SURROGATE AGONIST CAN BE BEND
INTO EFFECTOR MEMORY BIAS,
EFFECTOR CENTRAL MEMORY BIAS AND
THIS JUST REALLY WAS JUST A
REALLY FASCINATING RESULT, I
THOUGHT BECAUSE THIS IS DUE IN
LARGE PART TO SIMPLY THE FACT
THAT COMPARED TO IL2, HERE'S THE
NATIVE IL2 RECEPTOR GEOMETRY,
THESE SURROGATE AGONISTS ARE
DIMERIZING THE RECEPTORS IN
DIFFERENT SHAPES AND DIFFERENT
DISTANCES BETWEEN THE
INTRACELLULAR DOMAINS IN VERY
UNPREDICTABLE WAYS THAT HAVING
THESE KINDS OF EFFECTS ON
SIGNALING AND FUNCTION.
SO EISN'TIALLY THIS A WAY OF
TAKING A SINGLE CYTOKINE AND
CREATING, YOU KNOW A HUGE NUMBER
OF DIFFERENT FLAVORS OF THAT
CYTOKINE THAT HAVE DIFFERENT
FUNCTIONS.
AND SO WE'RE DOING THIS IN A LOT
OF DIFFERENT SYSTEMS NOW, EACH
VARIANT MIGHT HAVE A DIFFERENT
PLACE IN, IMMUNE DISEASE, A
DIFFERENT THERAPEUTIC
INDICATION.
WE'VE ALSO DONE THIS WITH TYPE 1
INTERFERON AGONISTS, WHERE AGAIN
WE MADE NANO BODIES AGAINST THE
EXTRA CELLULAR DOMAINS,
[INDISCERNIBLE] AND DID A MATRIX
AGAINST ALL AND HERE WE FOUND A
MUCH LOWER HIT RATE OF AGONISTS
SO INTERFERON RECEPTOR SEEMS TO
BE MUCH MORE STRUCTURALLY
RESTRICTIVE IN ITS ORIENTATION
THAN IL2 RECEPTOR BUT
NEVERTHELESS WE FOUND SOME HITS
AND JUST SHOWING YOU HERE NOW,
PARTIAL AGONISTS COMPARED TO
TYPE 1 INTERFERON AND A459 CELLS
AND EVEN THOUGH THEY'RE PARTIAL
AGONISTS THEY'RE JUST AS POTENT
AND TYPE 1 INTERFERON FOR
EXAMPLE, IN INHIBITING
SARS-COV-2 REPLICATION IN A549
CELLS.
THESE ARE DOSE RESPONSE CURVES
FOR INHIBITING SARS-COV-2
REPLICATION AND IN WORKING WITH
THE [INDISCERNIBLE] LABS WE
FOUND THEY INHIBIT OTHER TYPES
OF VIRUSES, LIKE ZIKA VIRUS EVEN
MORE POTENTLY THAN TYPE 1
INTERFERON.
BUT THE REAL PART OF THESE
MOLECULES THAT I FIND
INTERESTING IS THAT 1 OF THE
LIMITATIONS OF USING TYPE 1
INTERFERONS IN THE CLINIC THEY
HAVE A LOT OF SIDE EFFECTS,
CALLED FLU-LIKE SED EFFECTS THEY
ACTIVATE A LOT OF
PROINFLAMMATORY GENES.
AND OUR SURROGATE ACTIVITIES
BARELY AGGRAVATE THEM, THEY'RE
BARELY ATTENUATE SO OUR
PREDICTION IS THEY WILL HAVE
MUCH LESS PRO INFLAMMATORY
ACTIVATION, SO AND I STARTED A
COMPANY TO STUDY THESE IN
ANTIVIRALS WITH LESS SIDE
EFFECTS.
I WANT TO GO HERE FROM NOW, SO
UNTIL NOW WE'VE BEEN TALKING
ABOUT KNOWN CYTOKINE SIGNALING
PATHWAYS, TUNING DOWN SYSTEMS
WITH DIFFERENT KINDS OF
MOLECULES.
WE'RE ALSO THINKING ABOUT THIS
IN A DIFFERENT KIND OF WAY,
WHICH IS BRINGING TOGETHER
CYTOKINE RECEPTORS THAT DON'T
NORMALLY PAIR ON THE CELL
SURFACE, THROUGH INDUCED
PROXIMITY AND IF YOU THINK ABOUT
IT, ALL THE PLEIOTROPY THAT
WARREN HAS WRITTEN ABOUT AND
PUBLISHED ON SO PROLIFICALLY IS
ALSO AN OPPORTUNITY TO CREATE
NEW CYTOKINE SIGNALINGAXIS THAT
DON'T EXIST IN NATURE BECAUSE
EVERY TYPE OF IMMUNE CELL HAS
MULTIPLE DIFFERENT CYTOKINE
RECEPT ONORS AND YOU CAN THINK
OF THAT AS A MENU OF POSSIBLE
WAYS TO BRING THEM TOGETHER AND
YOU THINK OF IT, WE HAVE
SOMETHING LIKE 30 OR 40
DIFFERENT CYTOKINE RECEPTORS AND
THAT THE KINASES ASSOCIATED WITH
THE INTRACELLULAR DOMAINS HAVE
THE ABILITY TO CROSS TALK WITH 1
ANOTHER, IT REALLY OPENS UP A
WHOLE NEW POSSIBILITY OF
CREATING NEW CYTOKINE SIGNALING
AXIS.
SO HERE'S 1 EXAMPLE, PRINTS
BEING DONE BY [INDISCERNIBLE] IN
MY LAB WHERE WE TOOK THE IL2
RECEPTOR COMPLEX AND THE IL10
RECEPTOR COMPLEX AND WE BORROWED
A CHAIN FROM EACH COMPLEX TO TRY
TO MAKE A IL2 OR 10 USING THIS
NANO BODY TYPE OF APPROACH.
AND FOUND A NUMBER OF THITS THAT
ACTIVATE THEM AND SOME EARLY
FUNCTIONAL ASSAYS OF THIS HE
FINDS THAT DIFFERENT VERSIONS
ARE ABLE TO ACTIVATE CDHT-CELL
ACTIVATION AND ALTHOUGH NOT AS
POTENTLY AS IL2.
SO WHAT WE REALLY HAVE HERE IS
LIKE A NEW FLAVOR OF IL2.
BECAUSE WE REPLACED COMMON GAMMA
CHAIN WITH IL10 OR BETA, IT'S
GOING TO HAVE ITS OWN UNIQUE
CONSTELLATION OF SIGNALING
ADAPTORS THAT WERE IN THE
PROCESS OF EXPLORING BUT SHOW
THIS IS A CYTOKINE THAT DOESN'T
EXIST IN NATURE, BUT HAS
ACTIVITY ON NATURAL CELLS AND
SO, WE'RE LOOKING INTO THIS A
LOT DEEPER AND REALLY TRYING TO
UNDERSTAND WHAT NEW PROPERTIES
THESE SYNTHETIC CYTOKINES HAVE.
SO I WANT TO PIVOT A LITTLE BIT
HERE FROM SYSTEMIC CYTOKINES
WHICH OUR LAB IS MAINLY A
PROTEIN LAB AND SO WE'RE MORE
INTERESTED IN MAKING PROTEIN
MOLECULES THAT DO THINGS ON
CELLS, BUT CELL THERAPY IS ALSO
A GREAT OPPORTUNITY FOR PROTEIN
ENGINEERING AND I'M GOING TO
SHOW YOU WHAT OUR CONTRIBUTION
TO ADVANCING CELL THERAPY HAS
BEEN AND THAT'S BEEN IN
ORTHAGOREAN NOLL IL2 SYSTEM.
SO WHAT I'M SHOWING YOU HERE IS
THE IL2 RECEPTOR COMPLEX AGAIN.
AND SO, YOU KNOW 1 OF THE
LIMITATIONS OF CELL THERAPY IS
THAT WHEN YOU GIVE PEOPLE
ENGINEERED T-CELLS THEY DON'T
EXPAND AS MUCH AS YOU WOULD LIKE
THEM TO AND YOU WOULD LIKE TO
GIVE THEM IL2 TO EXPABD THOSE
CELLS BUT IL2 IS VERY TOXIC.
SO IF WE COULD MAKE AN IL2 THAT
WAS SELECTIVE FOR IL2 RBETTA
THAT WOULD BE 1 POSSIBLE WAY OF
SOLVING THAT PROBLEM SO WHAT WE
DID WAS WE MADE A MUTANT IL2
THAT NO LONGER BOUND TO
WILD-TYPE IL2 RBETTA AND A
MUTANT BETA THAT NO LONG BOUND
TO ILR2, BUT THEY BOUND TO EACH
OTHER AND THE IDEA BEING THAT
YOU COULD INSTALL THAT INTO
ENGINEERED T-CELLS AND PUT THOSE
CELLS INTO PEOPLE OR MICE, DOSE
WITH THE MUTANT CYTOKINE AND GET
SELECTIVE EXPANSION OF THE
ENGINEERED T-CELLS.
AND IN FACT, IT'S WORKED
BEAUTIFULLY.
AND THESE ARE THE DIFFERENT
THINGS CAN YOU DO WITH IT.
YOU CAN EXPAND CELLS, WE CAN
TRANSPLANT WITHOUT HOST
PRECONDITIONING, THERE'S REDUCED
TOXICITY MITIGATING EXHAUSTION
AND A LOT OF REASONS WHY THIS IS
A VERY POWERFUL TECHNOLOGY AND
JUST TO SHOW YOU A LITTLE BIT
ABOUT HOW WE DID IT, THIS IS
ORIGINAL ENGINEERING WAS DONE BY
JONATHAN [INDISCERNIBLE] IN MY
LAB, AND THE WAY HE WENT ABOUT
IT WAS MAKING A MUTATION IN THE
RECEPTOR SO IT
ABNORMALITIESIGATED IL2 BINDING
AND THIS IS BASED ON THE
STRUCTURE AND THEN CREATING A
LIBRARY ON IL2 AND SELECTING
AGAINST THE MUTANT, SELECTING
FOR THE MUTANT AND AGAINST THE
WILD-TYPE RECEPTOR.
SO HERE'S WHAT THIS LOOKS LIKE,
BASICALLY, THIS IS NOW THE
LIBRARY BINDING TO IL2 R, THE
MUTANT RECEPTOR THAT DOESN'T
BIEBD TO IL2 AND YOU CAN SEE
NEITHER OF THEM ARE BINDING
REALLY WELL BUT AFTER THE
SELECTION, YOU GET SELECTIVITY
FOR THE MUTANT RECEPTOR.
VERSUS THE WILD-TYPE RECEPTOR SO
NOW WE HAVE, WHAT WE CALL AN
ORTHOGONAL IL2.
AND WHAT THIS LOOKS LIKE IS THAT
WHEN YOU UPON TRANSDUCE T-CELLS
WITH THE ORTHOBETTA RECEPTOR AND
THEN ADD THE ORTHOCYTOKINE YOU
GET A STAT 5 SIGNAL WHEREAS WHEN
YOU APPLY THE ORTHOGONAL
WILD-TYPE T-CELLS WE DON'T GET
ANY GROWTH IN THIS CASE.
WE DEMONSTRATED THIS IN VIVO,
JONATHAN AND LEON IN MY LAB HAVE
DONE EXPERIMENTS WHERE WE CAN
MIX THE ORTHO2 CELLS WITH THE
NORMAL T-CELLS AND WE GET VERY
SPECIFIC EXPANSION OF THE
ORTHOT-CELLS BY THE
ORTHOCYTOKINE.
AND THAT IS NOT--WE DO NOT SEE
EXPANSION OF CELLS THAT DO NOT
HAVE THE ORTHOGONAL RECEPTOR.
AND WE'VE SHOWN WITH MIKE MALONE
HERE, HERE'S 1 EXAMPLE IN A
CAR-T MODEL, IN A CD11 KARKS-T,
CELL AND WE PUT THEM INTO THE
CAR-T CELLS AND THEN WE SEE DOSE
DEPENDENT EXPANSION OF THE
T-CELLS AT DIFFERENT DOSES OF
THE ORTHOGONAL CYTOKINE, SO THIS
IS 2 LONG EXPANSION OF THESE
CELLS.
AND IN THIS EXPERIMENT, MIKE
SHOWED VERY NICELY THAT THE CAR
T WITHOUT THE ORTHOGONAL IS
CONTROLLING THE TUMORS BUT WHEN
YOU ADD THE ORTHOGONAL CYTOKINE,
YOU GET THE DOSE TO DOSE
SIGNIFICANTLY FEWER CELLS.
FOR EXAMPLE, THIS IS 200,000
CELLS, IT'S A BIT EFFECTIVE AND
IN HALF A MILLION CELLS WHICH
JUST AS EFFECTIVE AS A MILLION
CELLS, SO IT ALLOWS YOU TO PUT
IN MANY FEWER CELLS INTO THE
MICE AT ALL SO THE SURVIVAL
BENEFIT IS CLEAR AND THIS IS
ALSO ANOTHER MOLECULE WHICH IS
GOING INTO THE CLINIC THROUGH
SYNTH KINE.
NOW THERE'S BEEN A RECENT TWIST
WITH THIS ORTHOBETTA SYSTEM I
WANT TO TELL YOU ABOUT AND THIS
IS A COLLABORATION BETWEEN MY
LAB TONY, KARL, AND
[INDISCERNIBLE] WHO'S NOW HERE
AT STANFORD AND WHAT THIS SYSTEM
DOES, IT AFFORDS YOU THE
POSSIBILITY OF SWAPPING OUT THE
INTRACELLULAR DOMAIN OF THE BETA
RECEPTOR SO ANY OF THE COMMON
FAMILY AND HERE'S A PICTURE FOR
WARREN'S REVIEWS ON ALL THE
COMMON GAMMA CHAIN CYTOKINES SO
THIS INSPIRES US TO REPLACE THE
BETA WITH EACH OF COMMON GAMMA
CHAIN CYTOKINE.
AND THIS CHES AN INTERESTING
EXPERIMENT, SO THIS IS SHOWING
YOU ALL THE ICDs ACTIVATING
STAT 5, 3 AND EASY AND 6, SO
PREDICTABLY THE ORTHO4 RECEPTOR
ONLY ACTIVATES STAT 6 BUT ORTHO9
ACTIVATES 1, 3, AND 5.
SO IT'S ALMOST ACTIVATING ALL
THE STATS EXCEPT STAT 6.
THE OTHERS ARE INTERESTING TOO,
ORTHO7 IS A PARTIAL AGONIST FOR
STAT 5, BUT WE PUBLISHED IN
PAPER LAST YEAR [INDISCERNIBLE]
IS THE FIRST AUTHOR WORKING WITH
LEON AND CANNED CANNED--[INDIS]
IN KARL'S LAB BUT I WILL JUST
FOCUS ON 9 FOR NOW BECAUSE WHAT
WE ALSO FOUND WAS THAT IF WE
LOOK AT EFFECTOR MARKERS LIKE
CD62, AND STEMNESS MARKERS LIKE
FAST AND SCA, ORGT O 9 AND ORGT
O 21 SEEM TO HAVE ADVANTAGES
OVER THE COMMON FAMILY MEMBERS
ESPECIALLY WHEN IT AM CANS TO
STEMNESS, SO WE CHOSE ORTHO9
RECEPTOR TO GO DEEPER INTO THE
STUDIES AND THIS IS
[INDISCERNIBLE] WORK AND SHE
SHOWED THAT ORTHO9 IS ABLE TO
HAVE ADOPTED CELL IN THE ABSENCE
OF LYMPHDEPLETION.
AND NOT ONLY THAT BUT YOU SEE
BETTER INFILTRATION OF THE
TUMORS, YOU SEE THE TISNI SPLOT,
THE ORTHO9 CELLS HAVE MORE OF AN
ACTIVATED PHENOTYPE THAN THE
ORGT O 2 CELLS AND THEY'RE ALSO
THE STEMNESS MARKERS, THIS IS
ORGT O 9 RIGHT HERE, THE
STEMNESS MARKERS ARE UP AND
REDUCE EXHAUSTION MARKERS AND
THINGS LIKE THAT SO THIS WAS
REALLY SURPRISING, I MEANI IL9,
REALLY WE DON'T KNOW A LOT ABOUT
IL9, OR WHAT IT'S DOING OR THERE
AREN'T MANY T-CELL RESUPPORTORS
THAT RECEPTIVE TO IL9 BUT WHEN
WE TAKE IT AND PUT IT IN A NEW
CONTEXT IS REVEALS THESE
FUNCTIONS WE IN NO IDEA ABOUT
AND THIS IS DATA FROM KARL'S LAB
WHICH SUPPORTED CANNED WHAT DATA
IN THIS CHIMERIC--CAR T SOLID
TUMOR AND YOU KEY IS IN THIS
[INDISCERNIBLE] EXPERIMENT, HERE
ARE THE CAR Ts THAT ARE
SUPPORTED BY ORTHO2 AND HERE
THEY ARE WITH ORTHO9 SO THEY'RE
MUCH BETTER CYTOTOXICITY BY THE
ORTHO9 AND THEY'RE ALSO ENHANCED
EFFECTOR FUNCTIONS AND ALSO THE
STEMNESS AND THIS IS NOW IN A
SOLID TUMOR MODEL SO REALLY THE
ORTHO9 SYSTEM IS SUPER
INTERESTING AND WE'RE LOOKING
DEEPER INTO IT AS PERHAPS THE
NEXT GEN BEYOND ORTHO2, AND--BUT
THERE'S 1 LIABILITY OF THE
ORTHO9 SYSTEM COMPARED TO ORTHO2
WHICH IS ORTHO2 THE CELLS EXPAND
TREMENDOUSLY IN THE SYSTEMIC
CIRCULATION, ORTHO9 DOES NOT
HAVE A STRONG PROLIFERATIVE
SIGNAL SO WHAT LEON SUH HASSA
BEEN DOING IN MY LAB IS SHE'S
TAKING A HARD LOOK AT THE IL9
RECEPTOR INTRANSLATIONAL
RESEARCH CELLULAR DOMAIN
TYROSEENS AND HE'S BEEN MUTATING
THEM AND DOG PROTEIN ENGINEERING
ON THE EXTRA CELLULAR DOMAIN AND
YOU CAN SEE WHENY WOO BEING
LOAMACY THEA THE STAT 5
SIGNALING, STAT 5, 3, AND 1,
DON'T FOCUS IN ON THE DETAILS
HERE, JUST LOOK AT HERE'S ORTHO2
VERSUS ORTHOWILD-TYPE AND YOU
CAN SEE IT LOOKS LIKE AS WE'VE
SHOWN EARLIER ORTHO9 ACTIVATES
ALL OF THEM BUT LEON HAS MADE
VERSIONS WHICH NOW CONTUNE THE
DEGREE OF STAT ACTIVATION OF THE
DIFFERENT STATTINGS AND AT THE
SAME TIME OF RESCUING THE ASSAYS
HE'S BEEN ABLE TO RESCUE THE
ILOORKS 9, IT'S NOT AS GOOD AS
IL2 BUT IT'S STILL SOMETHING
WHILE ALSO PRESERVING THE
STEMNESS PROPERTIES TO DIFFERENT
DEGREES.
AND SO WE'RE BEGINNING TO WORK
WITH MIKE MILONE AND THE SAME
TESTING AND TUMOR VARIANTS ABOUT
ASKING WHAT IS REALLY THE BEST
COMBINATION OF SIGNALS HERE FROM
THE 2 MUTANTS HERE AND THE
TUNABILITY FOR THERAPEUTICS.
I'LL VIEW THIS SECTION HERE, ALL
THESE CELL FATE DECISIONS ARE
MADE IN THE CONTEXT OF A
CYTOKINE MILIEU AND WE THINK
THAT THESE EXPERIMENTS REALLY
OPEN UP A LOT OF POSSIBILITY OF
TUNING CYTOKINES EARLY IN THESE
FATE DETERMINE NATION PATHWAYS
AND POSSIBLY CREATING NEW CELL
FATES OR ALTERING BAIT CHOICES
IN ADDITION TO THE
DIFFERENTIATED CELL TYPES I'VE
TALKED ABOUT SO FAR.
SO THAT'S KIND OF WHERE WE'RE
GOING, I WANT TO FINISH UP HERE
GETTING BACK TO MECHANISM, YOU
KNOW AFTER A LONG, YOU KNOW I
STARTED WORKING ON THE JACK
STRUCTURE WHILE I WAS A POST DOC
AT SCRIPS AND FINALLY IN 2022,
THROUGH CRYOEM, A TEAM OF PEOPLE
IN MY LAB, CALEB, KEVIN,
RECENTLY, NATHANIEL, WE MANAGED
TO STRUCTURE A FULL LEJT JACK
AND SO IN THIS STRUCTURE IN THE
1 IN THE LEFT HERE, THE KINASE
DOMAINS HERE WERE A PART ASK WE
KNOW THAT WHEN THE JACK--SO THIS
IS THE INTRACELLULAR DOMAIN.
WHEN IT BINDS AND KIND OF--THE
CYTOKINE BRINGS THESE TOGETHER,
THESE KINASE DOMAINS HAVE TO
TRANSPHOSPHORYLATE TO ACTIVATE
TURGTER SIGNALING SO RECENTLY,
NATHANIEL AND BOBBY SAX TON AND
MY LAB, [INDISCERNIBLE] MANAGED
TO TRAP AIAN CONFIRMATION OF THE
CLUSTER DOMAINS WHERE THEY'RE IN
COMMUNICATION WITH EACH OTHER.
SO NOW THIS IS THE ACTIVE
TRANSFORFORALATING POSE OF THIS
1 AND THE MESSAGE HERE THERE'S A
LOT OF FLEXIBILITY IN THESE
KINASE DOMAINS AND YOU WOULD
IMAGINE THERE HAS TO BE BECAUSE,
YOU KNOW THESE INTRACELLULAR
DOMAIN TAILS KIND OF HANG OFF
HERE AND KIND OF WIND THEIR WAY
DOWN TO THE KINASE DOMAINS AND
THE KINASE DOMAIN VS TO BE ABLE
TO ACCESS DIFFERENT TYROSEENS ON
THE DIFFERENT CELLULAR DOMAINS
AND THEY NEED TO BE SOMEWHAT
MOBILE AND SO WE'RE CONTINUING
TO ESCALATE OUR STRUCTURAL
STUDIES NOW TO TRAP A STAT
COMPLEX WITH THE JACK.
BUT WHAT I WANT TO ALSO SHOW YOU
TODAY IS WHAT WE LEARNED ABOUT
ONCOGENIC MUTATIONS FROM THE
STRUCTURE.
SO THERE'S THIS CLASSIC MUTATION
IN JACKS CALLED V17 F WHICH
RESPONSIBLE FOR MANY CLASSES,
IT'S A CONSTITTATIVE ACTIVATING
MUTATION IN THE PSEUDOKINASE
DOMAIN AND THIS IS A FIGURE FROM
THE ORIGINAL PAPER, APOLOGIES I
DON'T HAVE A REFERENCE HERE BUT
THIS IS SEVERAL GROUPS DETERMINE
THIS ABOUT THE SAME TIME ABOUT
15 YEARS AGO AND THIS MUTATION
MAPS RIGHT INTO THIS DIMER
INTERFACE RIGHT THERE BETWEEN
THE PSEUDOKINASE DOMAINS AND
IT'S AVAILING TO
[INDISCERNIBLE], SO WHAT IT DOES
IF YOU LOOK AT THE WILD-TYPE
INTERFACE BETWEEN THE
PSEUDOKINASE DOMAINS, IT'S A
VERY POORLY PACKED INTERFACE AND
SO IN THIS MODEL, THE CYTOKINE
IS REALLY RESPONSIBILITY FOR
BRINGING THE 2 RECEPTORS WITH
THEIR JACKS TOGETHER, TO FORM IN
DIMER.
HOWEVER, WHENEVER THE
VALENTINEDENT IS MUTATED TO A
[INDISCERNIBLE] LIKE IN V617 F
AND IT FILLS THE GAPS AND
IMPROVES THE SHAPE
COMPLEMENTARITY BETWEEN THE
DOMAINS AND WHEN THAT HAPPENS
NOW, THE JACKS ARE ABLE TO
DIMERIZE WITHOUT THE REQUIREMENT
OF THE CYTOKINE BRINGING THE 2
RECEPTORS TOGETHER.
SO IT'S ESSENTIALLY AN
ONMECHANISM MUTATION THAT JUST
SERVES TO CEMENT THIS DIMER
TOGETHER IN A CYTOKINE
INDEPENDENT WAY.
AND WE DID SOME EXPERIMENTS WITH
[INDISCERNIBLE] USING A SINGLE
MOLECULE TURF SEVERAL YEARS AGO
AND [INDISCERNIBLE] AND HIS
GROUP SHOWED THAT THIS
CONSTITTATIVE HOMER
DIMERIZATION, THE ABSENCE OF
LIGAND IS HOW B617 F WORKS SO
FOR INSTANCE HERE'S TFO RECEPTOR
AND EPO RECEPTOR ASK HAD IS JUST
ANOTHER WOR FORWARD
DIMERIZATIONOT CELL SURFACE AND
YOU CAN SEE WHEN THE WILD-TYPE
JACK IS MUTATED BOTH RECEPTORS
NOW DIMER ICE IN A LIGAND
INDEPENDENT FASHION.
SO NOW WITH THE STRUCTURE WE'RE
ABLE TO KIND OF PUT ALL THIS
TOGETHER, AND THIS LIGAND
INDEPENDENT DIMERIZATION IS DUE
TO THIS INCREASED SHAPE
COMPLEMENTARITY AS A RESULT OF
THIS MUTATION WHICH FORMS KIND
OF A GLUE BETWEEN THE JACKS TO
HOLD THEM TOGETHER AND THERE'S
ALL KINDS OF OTHER MUTATIONS IN
JACKS OUTSIDE OF THIS B617 F
THAT ARE ALSO ACTIVATES THAT
DON'T WORK THROUGH DIMERIZE AND
THIS IS ANOTHER FIGURE FROM THE
PAPER WHERE HERE WE HAVE THESE
CONSTITTATIVE ACTIVATING JACK
MUTATIONS, THEY DON'T CAUSE
DIMERIZATION AND SO WE'RE REALLY
INTERESTED IN THIS AND YOU KNOW
THESE JACK PAPERS
[INDISCERNIBLE] THERE AT NIH AND
CONGRATULATIONS TO JOHN, HE WAS
JUST SELECTED TO THE NATIONAL
ACADEMY.
JOHN REALLY PILOT PROJECT OR
NEARED THE DEVELOPMENT OF THESE
JACK INHIBITORS TO INHIBIT THESE
KINASE DOMAINS AND TREAT MILE O
PROLIFERATIVE NEOPLASMS, WELL
THAT'S LIKE A STANDARD OF CARE
IN THE CLINIC NOW, IT'S A PILLAR
OF THERAPY.
WE THINK ABOUT OTHER WAYS TO DO
THIS AND 1 WAY IS THROUGH
RECRUITING PHOSPHATASES TO THE
RECEPTOR, SO IN THIS CASE, THE
B617 F WOULD BE SIGNALING
CONSTITTATIVELY, IF WE CAN BRING
A FOZ TO TAISS--SYNTH TO THE
RECEPTOR AND HAVE THE
INTRACELLULAR DOMAIN ON THE
FOSTER NURSED TO TAISS--SYNTH
DIRECTLY DEPHOSPHORYLATE THE
RECEPTOR INSTEAD OF DIRECT
INHIBITION OF THE KINASE, WE
MIGHT GET MORE SPECIFICITY
BECAUSE YOU DO THIS WITH A
BI-SPECIFIC ANTIBODY SPECIFIC TO
THE RECEPTOR, SPECIFIC TO THE
SPECIFIC RECEPTOR AND SO THAT'S
KIND OF 1 THING WE'RE WORKING ON
THAT I WILL LEAVE YOU WITH AND
PRELIMINARY DATA THAT WE CALL
THE RIPPER MOLECULE IS ABLE TO
INHIBIT PROLIFERATION OF CELLS
BEARING THIS JACK 2 BF MUTATION,
YOU KNOW ALMOST EFFECTIVELY, AS
GOOD AS THE JACK INHIBITOR SO
THIS IS A PROOF OF CONCEPT
EXPERIMENT THAT WE CAN
DEPHOSPHORYLATE THAT JACK.
AND YOU KNOW WE'RE JUST SORT OF
THE BEGINNING HERE OF TRYING TO
EXPLAIN THIS AS A NEW WAY OF
INHIBITING THE ONCOGENIC
RECEPTORS.
SO I WILL END HERE AND JUST, YOU
KNOW TELL YOU THAT WE'RE 20
YEARS LATER, MY LAB IS MORE
EXCITED ABOUT CYTOKINE RECEPTORS
THAN EVER.
THIS IS KIND OF OUR MENU OF
CANDIDATES HERE WHEN YOU THINK
ABOUT THE DIVERSITY OF THE EXTRA
CELLULAR DOMAINS, THEIR TISSUE
EXPRESSION--CYTOKINE RECEPTORS
JUST LIKE ARTISTS HOW THE COLORS
THAT WE CAN MIX AND MATCH AND
DIFFERENT COMBINATIONS AND
CREATE NOW SHADES AND NEW HUGHES
AND SO WE'RE HAVING A LOT OF FUN
WITH THAT NOW AND HOPEFULLY WE
WILL HAVE NEW THINGS TO TELL YOU
ABOUT SOON.
SO I WANT TO FINISH LIE
ACKNOWLEDGING ALL THE PEOPLE WHO
DID THIS WORK.
I TRIED TO MENTION NAMES ALONG
THE WAY, SO I DON'T REALLY NEED
TO GO INTO DETAIL ON THIS SLIDE,
YOU PROBABLY HAVE SEEN THE NAMES
MENTIONED HERE OR THEIR
PUBLICATIONS AND SO, I WANT TO
THANK ALL THESE PEOPLE FOR THEIR
CONTRIBUTIONS OVER THE YEARS AND
ALSO AGAIN SYNTHEKINE IS
CREATING MOST OF THE MOLECULES I
SHOWED TODAY.
SO I WILL STOP HERE AND HAPPY TO
TAKE ANY QUESTIONS.
>>HEY, CHRIS.
THANK YOU VERY MUCH FOR FACAS
WONDERFUL TALK.
LET ME REMIND PEOPLE TO SEND
LIVE FEEDBACK WITH QUESTIONS SO
THAT WE CAN SEE THEM IN THE
CHAT.
LET ME START WITH 1 QUESTION,
CHRIS, BESIDES IL2, AND MAKING
AN ORTHOIL2 MOLECULE DO YOU HAVE
TD SENSE THAT OTHER
ORTHOCYTOKINES WOULD BE OF VALUE
TO CREATE SOMETHING OF A
PARALLEL SYSTEM WHERE YOU WOULD
PUT IN A DIFFERENT CYTOKINE
RECEPTOR AND HAVE THE DIFFERENT
PAIRED CYTOKINE.
SO NOT OVERLAPPING ORTHOCYTOKINE
SO CAN YOU SELECTIVELY ACTIVATE
2, 2 DIFFERENT PATHWAYS.
>>YEAH.
>>EIGHTY-THREE, YEAH,
ABSOLUTELY ORTHOGONALITY, I
THINK PEOPLE ARE WAKING UP TO
ORTHOLINALLITY AND ALL THE
THERAPEUTIC IMPLICATIONS IT HAS.
YOU KNOW ORTHOGONALITY IN
PRACTICE MADE IT SOUND DIFFICULT
BUT IT'S VERY DIFFICULT TO,A
CHIEF TO FIEBD A PROTEIN THAT
WILL HAVE NO ACTIVITY TO ANY
EXISTING PROTEIN IN THE BODY SO
IT'S NONTRIVIAL TO MAKE
ORTHOGONAL INTERACTIONS BUT IF
YOU CAN IT'S POWERFUL SO WHAT
WE'RE DOING NOW, I HAVE A NEW
POST DOC IN MY LAB, HE IS TRYING
TO MAKE THE DOUBLE ORTHO, NOT
JUST ORTHO AGAINST BETA BUT
ORTHO AGAINST GAMMA.
SO INSTEAD OF BEING LIMITED TO 1
ICD-CHANGE WE CAN NOW LOOK AT
ALL THE DIFFERENT THE CYTOKINE
RECEPTORS.
>>THIS THE CHA THE THERE ARE 2
QUESTIONS THAT ARE THE SAME,
WHAT ABOUT THE IMMUNO
GENERATEDISSITY RELATED TO THE
VARIOUS MODIFIED CYTOKINES.
>>RIGHT.
WELL, THAT'S A GOOD QUESTION,
AND IS DEFINITELY THE IT'S
DEFINITELY TOP OF MIND WHEN YOU
GO INTO THE CLINIC ON THESE
THINGS BUT I THINK WE'RE
STARTING TO--THERE ARE A LOT OF
CLINICAL TRIALS GOING ON RIGHT
NOW WITH DIFFERENT VERSIONS OF
ENGINEERED CYTOKINES, AND I
REALLY CAN'T SAY ANYTHING ABOUT
OUR PARTICULAR CYTOKINE AT THIS
TIME BUT IT'S TURNING OUT TO BE
NOT AS BIG OF A PROBLEM AS 1
MIGHT HAVE IMAGINE THAT IMMUNO
GENERATEDISSITY IS NOT ALWAYS
DUE TO MUTATIONS.
IT'S MOST OFTEN DUE TO THE
BEHAVIOR OF THE PROTEIN THAT
YOU'RE PUTTING INTO PEOPLE AND
SO, VERY WELL BEHAVED PROTEINS
THAT HAVE MUTATION TEND TO BE
PRETTY WELL TOLERATED.
>>SOMEONE ASKED HOW'S ORTHOIL2
DIFFERENT FROM IL15 IF IT IS IN
TERMS OF SIGNALING AND OVERALL
BIOLOGICAL EFFECTS?
>>YEAH, SO I DIDN'T GET INTO
THAT BUT THE DICHOTOMY BETWEEN
IL2 AND 15 IS INTERESTING.
SO, AND WE PUBLISHED ON THIS
WITH WARREN, YOU KNOW THEY USE
THE SAME RECEPTORS SO IL15 AND
IL2 USE IL2 R BETA AND COMMON
GAMMA CHAIN AND THEY SIGNAL
VERY, VERY--IN A VERY SIMILAR
WAY AND SO, NOW ORTHO2 DIFFERS
FROM IL15 IN THAT IT'S
COMPLETELY SPECIFIC FOR THE
ORTHAGOREAN NOLL RECEPTOR
WHEREAS IL15 IS SPECIFIC FOR THE
NATURAL RECEPTORS JUST LIKE IL2.
SO IL15 TO ME IS KIND OF ANOTHER
WAY OF DELIVERING AN IL2 LIKE
SIGNAL, BUT IT DOESN'T SOLVE THE
SPECIFICITY ISSUE.
>>CHRIS, LET ME ASK YOU A JACK
RELATED QUESTION THAT I KNOW YOU
THOUGHT ABOUT.
NOT ALL JACKS ARE NATURALLY
PAIRED WITH EACH OTHER IN
RESPONSE TO CYTOKINES LIKE ALL
HOMODIMERIC RECEPTORS ACT
THROUGH JACK 2, THE COMMON GAMMA
CHAIN, USE JACK 1 AND JACK 3,
BUT NOT EVERY PAIR IS
PHYSIOLOGICAL USED AND SO TO
WHAT DEGREE IS IT THAT THEY WORK
BETTER VERSUS THAT THEY JUST
WEREN'T COMBINED IN THE WAY--THE
WAY YOU DEVELOPED IT?
>>IT'S A FASCINATING REQUESTY
THAT DRIVES A LOT OF THE WAY I
THINK ABOUT THIS.
I HAVE A SLIDE WHERE I SHOW THE
MISSING JACK [INDISCERNIBLE]
THAT NATURE DID NOT PROVIDE FOR
AND I SPENT A LOT OF TIME
THINKING ABOUT THAT.
ONE POSSIBLE INTERPRETATION IS
NATURE TRIED THOSE COMBINATIONS
AND FOR WHATEVER REASON, THEY
WERE NOT GOOD FOR THE HOST AND
THEY WERE EVOLVED AGAINST,
THAT'S A POSSIBILITY.
I DON'T REALLY BELIEVE THAT.
WE HAVE NOW CREATED USING
SYNTHEKINES THOSE NONNATURAL
PAIRINGS FOR INSTANCE THERE'S NO
JACK 2 OR THRESHOLD PAIR NOTHING
98 IRB.
THERE IS NO TICK 2 JACK 1.
AND SO, WE'VE CREATED THOSE
PAIRINGS ARTIFICIALLY USING THE
ORTHOGONAL RECEPTOR SYSTEM AND
THEY HAVE INTERESTING SIGNALING
PROPERTIES AND SO NOW WE'RE
TRYING TO MAKE LIGANDS TO
ENFORCE THOSE PAIRINGS AS WELL
BECAUSE I REALLY THINK THERE'S
SOMETHING, THERE'S SOMETHING
HAPPENING THERE THAT'S GOING TO
BE VERY INTERESTING.
>>THERE ARE MORE QUESTIONS IN
THE CHAT, APOLOGIES, I WASN'T
NAMING PEOPLE BUT RUSS
[INDISCERNIBLE] ASKS, I FOUND
THE ABILITY TO REENGINEER THE
CYTOPLASM PORTIONS OF ENGINEERED
ILRECEPTORS EXCITING DO YOU
EXPECT THAT THE ABILITY TO
CHANGE THE CYTO[INDISCERNIBLE]
EXTRA CELLULAR PORTION TO BE
WIDELY APPLICABLE WHAT ABOUT
OTHER TYPES IMMUNORECEPTORS.
>>YEAH, ABSOLUTELY, LIKE I SAID
WE WANT TO MAKE THE DOUBLE
ORTHO, SO WE CAN SAMPLE ALL THE
DISCIPLINARY MERRIC COMBINATIONS
INCLUDING NONNATURAL JACK
PAIRINGS AND IT'S BEEN DONE IN
OTHER SYSTEMS AS WELL.
I MEAN PHIL GREEN BERG WAS
MAKING SO CALLED SWITCH
RECEPTORS QUITE A FEW YEARS AGO
WHERE HE WAS MAKING CHIMERIC
RECEPTORS, YOU KNOW FOR
INSTANCE, GCSF, INTERRACELLULAR
DOMAIN, INTERLEUKIN 2 EXTRA
CELLULAR DOMAIN SO THE CONCEPT'S
BEEN OUT THERE AND AROUND FOR A
WHILE AND MAKE THEM COMPLETELY
ORTHOGONAL AND I THINK IN CELL
THERAPY AND ACTIVITIES AND
PROJECTS THEY--THE IS WHERE THE
ACTION IS RIGHT NOW IS USING
THESE TYPES OF SYSTEMS TO
DELIVER NEW SIGNALING TO CELLS.
>>THERE'S A QUESTION BY
[INDISCERNIBLE] NICE HIT ON
CANCER MODELS HAVE YOU TRIED
ENGINEERED IL2 AND OTHER
CYTOKINES IN HIV?
>>WE HAVE NOT.
WE HAVE NOT.
I'M PARTICULARLY INTERESTED IN A
NEW KIND OF IL2 WE MADE THAT HAS
WE BROUGHT IN A IL3 MODEL AND
THAT WE'RE PARTICULARLY
INTERESTED IN TRYING THAT, I
WANT TO TRY THAT IN MODELS WHERE
T-CELL EXHAUSTION IS HAPPENING
IN THE MODELS.
Y SO WE HAVEN'T TRIED IT BUT I'M
INTERESTED.
>>CAN A QUESTION, CAN YOU
EXPAPPEDOT BEHAVIOR OF THE
PROTEIN THAT IMPACT ON IMIEWP O
GENERATEDISSITY.
>>IT'S MAINLY AGGREGATED
PROTEINS.
PROTEINS THAT DON'T BEHAVE WELL,
THAT DOESN'T HELP YOU MUCH BUT
FOR A BIOCHEMIST, YOU SAY A
PROTEIN DOESN'T BEHAVE WELL, YOU
KNOW WHICH WAY THE WIND IS
BLOWING AND SO WELL BEHAVED
PROTEINS, YOU KNOW MONODISPERSED
REALLY HAPPY, WELL EXPRESSED
PROTEINS, TEND TO NOT BE AS
IMMUNOGENIC AS MORE AGGREGATED
PROTEINS AND IT'S NOT A HARD
SCIENCE, BUT THAT'S KIND OF WHAT
THE DATA'S SHOWING US.
>>OKAY, I THINK THIS IS REALLY
WONDERFUL AND RISK RECEPTION
AFTERWARDS BUT WE WILL HOPEFULLY
THANK YOU.
>>THANKS A LOT AND THANKS TO
EVERYONE FOR THE QUESTIONS.
>>THANK YOU.