THE BASIC PROGRAM HERE AT
DADS AND WE'LL HEAR FROM REACH
AND THE ACRONYM STANDS FOR
RESEARCH ENTERPRISE TO ADVANCE
THE CURATOR HIV.
THE CO-P.I.s ARE BRAD JONES AT
WEILL CORNELL AND BRAD IS IS
GOING TO GIVE THE OVERVIEW SO
NOW I WILL TURN IT OVER TO BRAD
AND -- OH, MARINA IS GOING TO
GIVE THE OVERVIEW.
>> I'LL BRIEFLY INTRODUCE --
GOOD MORNING, EVERYONE.
PLEASE COME IN.
I'LL BRIEFLY INTRODUCE OUR
COLLABORATORY.
THE STRUCTURE OF OUR LEADERSHIP
AS MENTIONED I CO-LEAD THIS WITH
BRAD AND IN OUR EXECUTIVE
COMMITTEE WE HAVE
REPRESENTATIVES FROM THE THREE
RX AND WE HAVE OUR PARTNERS AND
OUR COMMUNITY ENGAGEMENT TEAM.
THIS IS HAVE THE ORGANIZATION
STRUCTURE OF OUR COLLABORATORY
UNDER RF1.
WE HAVE BEEN APPLYING DIFFERENT
METHODS IN AN ATTEMPT TO DEFINE
THE RELATIONSHIP BETWEEN HIV
RESERVOIRS, T CELLS AND REBOUND
VIRUSES.
UNDER RF2 THE UNDER ARCHING GOAL
IS TO HARNESS THE CELLS TO
DIFFERENT IMMUNOLOGIC APPROACHES
AND BUILD THE DISCOVERY TO
TRANSLATION PIPELINE TRYING TO
UNCOVER MECHANISMS AND IDENTIFY
APPROACHES THAT CAN OVERCOME
BARRIERS TO ELIMINATING
RESERVOIRS.
THIS PAST YEAR WE TRIED TO
LEVERAGE THE FLEXIBILITY OF THIS
GRANT MECHANISM AND WERE ABLE TO
ALLOCATE FUNDS THAT ALLOWED US
TO SUPPORT HIGH PRIORITY
PROJECTS AND WE BROUGHT ON
INVESTIGATORS AND FOR EXAMPLE WE
WERE ABLE TO ALLOW MECHANISMS OF
CONTROL THROUGH THE STUDIES
CONDUCTED.
WE HAVE SUPPORTED A SERIES OF
PUBLICATIONS BUT PERHAPS WHAT
WE'RE MOST PROUD OF IS WE HAVE
ALSO TRIED TO FOSTER NEW
COLLABORATIONS, BRING IN NEW
INVESTIGATORS AND STRENGTHEN
EXISTING COLLABORATORS.
WHAT WE'RE TRYING TO ILLUSTRATE
IS OUR PROGRESS OVER TIME IN
BRINGING INVESTIGATORS TOGETHER.
WHERE YOU SEE THE LINES BECOME
THICKER AND THE KNOBS BECOME
DARKER REPRESENTING THE
DIFFERENT COLLABORATION.
RF1 IS LED BY THESE PEOPLE AND A
BIG COMPONENT IS OUR PARTNERSHIP
WITH THE HEALTH SCIENCES
PROGRAM.
AS MANY KNOW UGANDA WAS THE
EPICENTER OF THE HIV EPIDEMIC IN
UGANDA.
DESPITE ALL THE STRIDES MADE IN
TERMS OF HIV TREATMENT AND
PREVENTION STRATEGIES IT LEFT A
LARGE POPULATION OF PEOPLE WITH
HIV AND SIGNIFICANT SOCIAL
BURDENS.
SO TRYING TO BRING HIV CARE
EFFORTS TO THE REGION HAS BEEN
CHALLENGING.
ONE OF THE CHALLENGES BEING THAT
ONE COULD NOT RELY ON THE
ORIGINAL IPDA.
SO WE WORKED TO DEVELOP
SUCCESSFULLY WERE ABLE TO
DEVELOP THE NEW ADAPTED IPDA
WITH NEW PRIMERS AND PROBES THAT
FIT.
THIS TOGETHER WITH RESEARCH
FUNDED EQUIPMENT NOW SUPPORT
RESEARCH TO HAPPEN ON SITE FOT
ONLY BECAUSE OF THE EQUIPMENT
BUT BECAUSE RESOURCES WENT INTO
TRAINING OF DIFFERENT UGANDAN
ADVOCATORS REALLY ABLE TO CARRY
OUT RESERVOIR MEASUREMENTS IN
PARALLEL WITH RESPONSE
ASSESSMENTS LEVERAGING FROM
THEIR COHORT AND IN ANOTHER AREA
UNDER RF1 IS ADDRESSING THE
QUESTION OF HOW IS HIV
INTEGRATION AFFECTS PERSISTENCE
BY DIFFERENT METHODS AND WORK
CARRIED OUT AND YOU'LL HEAR MORE
ABOUT THIS IN THE NEW TALKS.
ONE SUCH METHODS COMES FROM THIS
STRATEGY THAT WAS DEVELOP TO
ISOLATE CD4 CELLS IN VIRUSES AND
NOT ONLY ISOLATED BUT CAN BE
EXPAN
EXPANDED IN VITRO AND NOW
THEY'VE BECOME VALUABLE TOOLS TO
INVESTIGATE LATENCY AND TEST
APPROACHES TO OVERCOME LATENCY
AND TRY TO UNDERSTAND AND
IMPROVE HOW IMMUNOLOGIC PRESSURE
TO ELIMINATE RESERVOIRS.
WE'RE TRYING DIFFERENT
IMMUNOLOGIC APPROACHES.
ONE OF THE THINGS WE INTEREST TO
LEVERAGE IS THE VARIOUS BNAB
STUDY WE CONDUCTED OR IN
PROGRESS AND ASK HOW THE
STRATEGY MAY INTERFERE WITH
DYNAMICS AND PERHAPS MEDIATE
CONTROL.
ONE SUCH STUDY WAS THE STUDY
YOU'RE ALL FAMILIAR WITH AND
TESTED BNAB DOING ART
INITIATION.
IF YOU REMEMBER FROM THE
PUBLICATION AMONG PARTICIPANTS
WHO HAD BEEN WITH THE VIRUS FROM
THE START OBSERVED IN BLUE IN
THE GRAPH ARE OBSERVED THE DELAY
IN REBOUND BUT ALSO THE BLUNTED
VIRAL REBOUND CURVES.
THIS ALSO INCLUDED ONE
PARTICIPANT WHO MAINTAINED FULL
VIRUS SUPPRESSION OVER FOUR
YEARS NOW.
THEY KNOW THE RESERVOIR IS
DYNAMIC AND INDUCIBLE.
INTERESTINGLY OVER TIME HAS
HIGHER PROPORTIONS OF THE VIRUS
IN THE HETEROCHROMATIN REGIONS
AND HAS POTENT CD 8 T CELLS.
TRYING TO UNDERSTAND CTL
PRESSURE SPECIFICALLY IN THIS
PARTICIPANT WHAT HE DID USING
THE MODELS HE HAD HE USED
SAMPLES AND TRANSFERRED INTO THE
MOUSE MODEL WHICH WAS FOLLOWED
BY REBOUND HERE IN THE GRAPH.
HE WAS ABLE TO THEN ISOLATE THE
REBOUND VIRUSES AND TRANSFER TO
NEW MICE.
WHAT THIS GRAPH SHOWS WITHOUT
ANY CD8 PRESSURE THE VIRUS GOES
UP AND HERE THE VIRUS IS IN BLUE
AND RISES AND STAYS UP AND AT
THE SAME TIME WHEN CD 8 T CELLS
ARE TRANSFERRED THERE'S A RISE
LEEKED TO A SIGNIFICANT EXTENT
AND SHOWS THE PRESSURE THE T
CELLS HAVE PUTTING ON AND WE CAN
DO THESE TYPES OF INTERVENTIONS.
HERE THE IDEA IS THROUGH A
SERIES OF DIFFERENT METHODS TO
HAVE CD 8 T CELL KILLING AROUND
HERE'S THE MOLECULES IDENTIFIED
WITH POTENTIAL TO DO SO AND BRAD
WILL TALK MORE ABOUT THE
INHIBITOR AND HOW IT HAS BEEN
SHOWN AT LEAST IN VIVO MODELS TO
ENHANCE CTL KILL.
WITH THAT I'LL PASS IT OVER TO
JESSICA.
>> I'M GO TO GO OVER QUICKLY
WHAT WE'VE BEEN DOING IN THE
COMMUNITY.
REACH IS FOCUSSED AROUND THE NEW
YORK CITY AREA AND LEAN INTO
THAT AND HAVE A LOCALIZED
PROGRAM AND TALK ABOUT WHAT
WE'VE BEEN DOING STATESIDE AND
WE HAVE HAD A WEBINAR SERIES AND
BEGUN TO LEVERAGE CAT INPUT IN
SEPTEMBER WE HELD A SPECIFIC
MEETING AND AROUND THE EVH AND
LOOKED AT HUMAN CRITICAL TRIALS.
THE CAB HAD AN EXTENSIVE PROCESS
TO LEARN WHAT THE DRUG IS AND
TAKE IT THROUGH THE HISTORY.
WE HAD DR. GRAMATTICA COME IN
AND LEAD PRESENTATIONS AND GO
THROUGH THE Q&A AND CAB ASKED
ABOUT THE DOSAGE, TOXICITY,
TISSUE SAMPLE AND HAS ANYTHING
BEEN DONE AND AT THE END OF THE
DAY DECIDED TO MOVE IT FORWARD
INTO THE -- THEY WOULD THINK IT
CAN MOVE INTO HUMAN TRIALS AND
SKIP NHP MODELS.
THE CAB IS DRAFTING A LETTER TO
THE BOARD AND IT'S LED BY OUR
TWO REPRESENTATIVES ROB WALKER.
WE'RE EXCITED TO SEE THIS MODEL
MOVE FORWARD AS THE CAB AS A
CONSULTATION.
I WANTED TO HIGHLIGHT SOMETHING
WE'RE EXCITED ABOUT AND THE IDEA
OF CONNECTING THE BENCH TO THE
CLINIC.
WE KNOW WE DON'T DO CLINICAL
TRIALS WITHIN THE STRUCTURE BUT
THERE'S ACTIVITY AND
COLLABORATION WITH CLINICAL
TRIALS AROUND AND WE HELD A
COMMUNITY EVENT IN AFRICAN
AMERICAN SERVICES IN HARLEM TO
REACH THE IMMIGRANT POPULATION
OR BLACK AMERICANS LIVING IN NEW
YORK.
WHERE HE HAD THE COORDINATOR
FROM THE HARLEM CENTER ADDS AN
AUDIENCE MEMBER.
WE HAD A FANTASTIC EVENT AND WE
HAD A BLACK TRANS FEMALE AND SHE
WAS ABLE TO COME UP.
HER INTERACTION AND ENGAGEMENT
LED TO HER INTEREST TO RESEARCH
AND LED HER TO CONNECT WITH
JONATHAN.
WHILE HE'S WELL VERSED IN THE
CLINICAL TRIALS WAS NOT AS WELL
VERSED IN THE COMMUNITY
ENGAGEMENT APPROACHES OR THE
LARGER LITERACY.
HE STARTED ATTENDING HER SUPPORT
GROUP.
I WORKED WITH JONATHAN TO
DEVELOP THE LITERACY TOOLS AND
MATERIALS TO WORK THROUGH THE
STRATEGIES AND ABLE TO CULTIVATE
THE LEADERSHIP THAT DOESN'T TO A
CLINICAL TRIAL FOR EXCLUSIONARY
REASONS BUT LED TO HER AND
SEVERAL MEMBERS OF THAT SUPPORT
GROUP ENROLLING IN TRANS CARE
STUDIES.
SO WE CAN REALLY SEE WHEN WE
ESTABLISH THESE LINKAGES WITH
THE COORDINATORS OF CLINICAL
CENTERS WE CAN BUILD BRIDGES AND
INVOLVE NEW INDIVIDUALS AND I
THINK THAT CAME UP YESTERDAY
HIGHLIGHTED BY THE COMMUNITY
ADVISORY BOARD THE NEED TO BRING
IN NEW VOICES AND THIS IS ONE
WAY IN THE I WANTED TO
ILLUSTRATE.
ANOTHER WAY TO BRING IN NEW
VOICES IS IT'S WORKED WELL WITH
THE INTERNATIONAL AIDS SOCIETY
TO HAVE AN INTERNATIONAL
ADVOCACY GROUP AND WE LIKE TO DO
THAT IN THE UNITED STATES AND
CURE WORKSHOP.
IN PARTNERSHIP WITH OTHER
COLLABORATORIES WE'RE LAUNCHING
A U.S. CURE WORKSHOP BASED ON
THE MODEL OF NATIONAL AIDS
SOCIETY.
PULLING CIVIL SOCIETY AND
ADVOCATES FROM AROUND THE UNITED
STATES ONE THE GOAL OF GETTING
THEM UP TO SPEED AND BRINGING IN
NEW VOICES INTO THIS
CONVERSATION.
SO WE WILL BE FOLDING THIS
FEBRUARY 24 THROUGH FEBRUARY 26
IN PHILADELPHIA NEXT YEAR.
APPLICATIONS WILL OFFICIALLY
LAUNCH NEXT WEEK AND EXCITED
ABOUT THE ABILITY TO SPREAD THIS
FAR AND WIDE AND PARTNER WITH
GROUPS TO DIVERSIFY THE
INDIVIDUALS COMING OUT WITH AN
EMPHASIS ON TRANSLATING SCIENCE
AND THE COMMUNICATION BEING
FORWARD.
I WANTED TO HIGHLIGHT TWO
MEMBERS.
THEY'RE NOT HERE TODAY BUT JOHN
SCHOEPP AND GAYLE DORSKY THEY'RE
NEW TO THIS WORK BUT HAVE DONE A
FANTASTIC JOB FACILITATING
CONNECTIONS WITH NEW YORK CITY
AND RECRUITMENT.
THEY HELPED FACILITATE OUR
JUNIOR MEMBER AND WE BROUGHT IN
MENTAL HEALTH TRAINING.
VVNTS AROUND HOMELESSNESS AND
KEEP US UP TO DATE WITH WHAT'S
FUN AND HAPPENING IN NEW YORK
CITY.
WANTED TO HIGHLIGHT THE AMAZING
WORK THEY DO AND I KNOW THEY'RE
ONLINE TODAY AND THANK YOU FOR
YOUR EFFORTS AND WITH THAT I'LL
HAND IT TO PAUL.
>> GOOD MORNING.
THE REASON WE'RE GATHERED HERE
FUNDAMENTALLY IS BECAUSE THERAPY
REVEALS THE PRESENCE OF A SMALL
SUB POPULATION OF VIRUSES THAT
ADOPTED A DIFFERENT LIFESTYLE
THAT IS REPLICATING NOT BY
CONVENTIONAL MECHANISMS BUT
PIGGY BACKING ON CELL DIVISION
MAKING THEM IMPERVIOUS AND I'M
TALKING ABOUT MODEL SYSTEM TO
ADVOCATE THE FACTORS THAT THE
GOVERNMENT DECISION OF A PRO
VIRUS TO ENTER THIS SORT OF
ALTERNATIVE STYLE OF REPLICATION
BY OTHER RETROVIRUSES AND IN
PARTICULAR WHETHER THE SITE OF
INTEGRATIONS INFLUENCES THE
DECISION AND POTENTIALLY
IMPACTED CELL DIVISION
PROPAGATION OF THE RESERVOIR.
SO THE MODEL SYSTEMS BASED IN
PART ON A REPORTER PROVIRUS THAT
MAINTAINS NATIVE MECHANISMS OF
HIV GENE REGULATION.
WE SIMPLY REPLACE ONE OF THE HIV
GENES WITH A REPORTER GENE THAT
PLACES A SURFACE MARKER ON CELLS
ENABLING THEM TO BE PURIFIED AT
LARGE SCALE INTO POPULATIONS
THAT HAVE ACTIVE OR LATENT
PROVIRUSES.
SO WHAT WE DO IS WE TAKE CELLS
FROM AND SUPPORT HIM HAD IN THE
VIRUS AND PURIFY THE INFECTED
CELLS AND INGRAFT MICE WITH
ABOUT 5 MILLION CELLS EACH
CONTAINING A PRESUMPTIVE INTACT
TRANSCRIPTIONALLY ACTIVE PRO
VIRUS.
THE CELLS THEN LIVE IN THE MOUSE
FOR MONTHS.
IMPORTANT TO RECOGNIZE THERE IS
NO CONVENTIONAL HIV REPLICATION
GOING ON HERE.
THERE IS NO IMMUNE SELECTION.
THE PROVIRUS IS FREE TO DECIDE
ITS OWN FATE UNENCUMBERED BY
SELECTION PRESSURES SO THE CELLS
GRAFT AND GROW AND THE
PROVIRUSES SOME MAINTAIN
EXPRESSION.
SOME GO TO A STATE OF LATENCY.
MONTHS LATER WE CAN RECOVER
DECENT NUMBERS OF HUMAN CELLS
FROM MOUSE TISSUES IN THIS
PARTICULAR EXAMPLE THE MAJORITY
OF THOSE CELLS HAVE LOST THE
REPORTER GENE EXPRESSION INTO
THE STATE OF LATENCY.
WE DEVELOPED AN ASSAY BASED ON
NUMEROUS PARALLEL INTEGRATION
SITE DERM DETERMINATIONS TO
QUANTIFY TO GENES.
WE FIND IN THE PRE-INFUSION
SAMPLE WE RECOVER MOST THE
INTEGRATIONS THAT BASED ON THE
NUMBER OF PROVIRUSES THAT GET
INTO THE ASSAY.
MOST OF THE INTEGRATIONS ARE IN
GENES.
HOWEVER IN THE MOUSE GRAFT WHILE
WE FIND THE EXPECTED NUMBER OF
TOTAL INTERGRANTS THE NUMBER OF
TOTAL ARE FAR FEWER BECAUSE
THERE HAS BEEN A VERY EXTENSIVE
DEGREE OF CLONAL EXPANSION OF
INDIVIDUAL CLONES IN THE MOUSE
GRAFT HETEROGENEOUS EXPANSION
SOME ARE GROWING BETTER THAN
OTHERS FOLLOWING ENDRAGRAFTMENT
THE MOUSE.
DOES THE SITE IMPACT THE ABILITY
TO PROLIFERATE IN PREFERENCE TO
OTHER CLONES IN THE GRAFT?
THE ANSWER IS TENTATIVELY, NO.
THOUGH THERE IS QUITE A BIT OF
OVERLAP IN THE GENES WE FIND
BEARING PROVIRUSES IN DIFFERENT
MICE, IF YOU LOOK AT THESE
CHARTS WHERE EACH DATA POINT
REPRESENTS AN INDIVIDUAL GENE
WITH AN INTEGRATED PRO VIRUS.
THE NUMBER OF MICE IN WHICH WE
FIND INTEGRATION TO ANY GIVEN
GENE IS NOT PREDICTED BY THE
ASSOCIATION OF THAT GENE WITH
HIGH LEVELS OF CLONAL EXPANSION.
ALSO, IF WE COUNT THE NUMBER OF
TIMES A GENE CARRIES A PROVIRUS
IN A MOUSE GRAPH MONTHS LATER,
THAT NUMBER IS QUITE STRONGLY
PREDICTED BY THE NUMBER OF
INTEGRATIONS WE HAVE IN THE VERY
SAME GENE IMMEDIATELY FOLLOWING
INFECTION WHERE THERE HAS BEEN
NO OPPORTUNITY FOR SELECTION.
WE SIMPLY DON'T FIND GENES WHERE
INTEGRATION IN THAT GENE FAVORS
SURVIVAL IN THE GRAFT ITSELF IN
THE MOUSE.
IN FACT, WHAT PREDICTS WHETHER A
GENE CARRIES A PRO VIRUS OR NOT
IS ITS SIZE AND EXPRESSION
LEVEL.
FACTORS WELL KNOWN TO FAVOR
INTEGRATION IN THE ABSENCE OF
ANY OTHER SELECTION QUESTION AND
THAT'S TRUE BOTH IN THE
PREINFUSION SAMPLE AND MONTHS
LATER IN THE MOUSE GRAPH.
SO QUITE STRONG EVIDENCE AT
LEAST IN THIS MODEL THAT THE HIV
PROVIRUS IS ACTING VERY MUCH AS
A PASSENGER AND NOT A DRIVER OF
CELL SURVIVAL OF CLONAL
EXPANSION.
SO TO QUESTION TWO, DOES THE
SITE OF INTEGRATION INFLUENCE
WHETHER THAT PROVIRUS IS GOING
TO ENTER LATENCY.
SO THE WAY WE ADDRESS THIS IS TO
HARVEST THE TISSUE AND APPLY
ARTIFICIAL SELECTION PRESSURE
WITH A MAGNET OR FLOW CYTOMETER
AND WHEN WE DO THAT WE SEE IN
THIS EXAMPLE MOUSE THE TYPICAL
PATTERN OF CLONAL EXPANSION BOTH
IN THE ACTIVE PROVIRUS
POPULATION AND IN THE LATENT
POPULATION.
KEY POINT THOUGH IS THE LATENT
POPULATION IS ALMOST
NON-OVERLAPPING.
FOR EXAMPLE THE PROVIRUS, THIS
CLONE WHICH REPRESENTS NEARLY
HALF THE LATENT PROVIRUS
POPULATION AND INCIDENTALLY AS
INTEGRATION IN A ZINC FINGER
GENE IN CHROMOSOMAL 19 IS
VIRTUALLY ABSENT FROM THE
POPULATION.
IN FACT IF YOU LOOK AT
PROVIRUSES AS A TOTAL ONE THAT
MAINTAIN ACTIVITY INTERFREIGHTED
MOST INTEGRATED MOST IN GENES
THAT ARE LATENT MOST ARE NOT IN
GENES.
A FEW ARE.
IF WE LOOK AT CHROMOSOME MARKS
AROUND ACTIVE AND LATENT
PROVIRUSES USING DATA FROM
ENCODE ANALYSIS OF MEMORY T
CELLS, LATENT PROVIRUSES ARE
ENRICHED FOR THE MARK ASSOCIATED
WITH HETEROCHROMATIN.
WE HAVE MODELS THAT SEEMS TO
REASONABLY WELL RECAPITULATE THE
PICTURES ASSOCIATED WITH THE
LATENT RESERVOIR IN HUMANS.
IN REACH WE HAVE THE UNIQUE
OPPORTUNITY TO COMPARE WHAT WE
DO IN OUR MODEL WITH CELLS THAT
COME FROM THE RESERVOIR.
FROM THE INGENIUS BRUTE FORCE
OF OUR LAB WE HAD THE CLONES AND
HAVE INTERESTING PROPERTIES AND
THE FACT THEY ALL HAVE AN INTACT
PROVIRUS FEW EXPRESS AND YOU SEE
THE CELL PROLIFERATING AND A
SMALL PERCENTAGE OF THEM MAKING
GAG AND THE PROCESS OF VIRAL
GENE EXPRESSION HAS BECOME
NEARLY COMPLETELY UNCOUPLED FROM
THE PROCESS OF CELL DIVISION AND
CELL ACTIVATION.
AND IF WE LOOK AT THE HISTONE
MARKS THIS IS FROM ANALYSIS AND
YOU CAN SEE THE SAME HISTONE
MARK AROUND THE AUTHENTIC
RESERVOIR CLONE.
SO ANOTHER PROPERTY THAT SEEMS
TO BE ASSOCIATED WITH THESE
AUTHENTIC RESERVOIR CLONES IS IF
YOU TREAT THEM WITH CONVENTIONAL
LATENCY REVERSING AGENTS AND
HERE'S A WHOLE PANEL WITH
DIFFERENT MECHANISMS OF ACTION
THEY ARE HAVE LITTLE TO ANY
EFFECT ON THE OF THE PRO VIRUS.
WE HAVE GAG AFFECTS THE TWO SUCH
CLONES WITH INTEGRATIONS AT
DIFFERENT -- THEY JUST DON'T
RESPOND WELL.
SO THERE'S OUR MODEL.
WE CAPITULATE LIKE THIS.
IT'S A LOT OF WORK TO ISOLATE
EVEN ONE CLONE FROM AN INFECTED
INDIVIDUAL BUT FROM OUR MICE WE
CAN IN PRINCIPLE DO THIS MANY
TIMES OVER.
WE'RE IN THE EARLY STAGES OF
DOING THAT SO I'M GOING TO SHOW
YOU A PILOT EXPERIMENT.
SO WHAT WE FIND IS IF WE GET 50
POSITIVE WELLS WITH HUMAN CELLS
GROWING FROM A COUPLE ANIMALS WE
FIND ABOUT A QUARTER OF THOSE
CLONES FROM ANIMALS AND ONLY
LATENT PROVIRUS.
UNFORTUNATELY IN THIS EXPERIMENT
WE UNDERESTIMATED THE EFFICIENCY
WITH WHICH OUR CLOTH PROCEDURES
WILL WORK.
MANY WELLS HAVE MORE THAN ONE
CLONING BUT HAVE NUMBERS WE
THINK THERE'S AN A SINGLE CELL
WITH ONE INTACT AND A MIXTURE
AND LATENT PROVIRUSES.
NOW IF WE TAKE ONE SUCH CLONE.
HERE'S AN EXAMPLE WHERE 15% OF
THE CLONES IS GFP POSITIVE AND
ADOPT A KITCHEN SINK APPROACH
AND THERE'S MANIPULATION TO GET
IT TO EXPRESS.
THAT'S TRUE OF SEVERAL CLONES
WHERE WE APPLIED THE PROCEDURE
AND WITH DIFFERENT STARTING
LEVELS OF EXPRESSION.
THEY SIMPLY DON'T RESPOND WELL.
AND SO TO SUMMARIZE AT LEAST IN
OUR MODEL, HIV INTEGRATION DOES
NOT SEEM TO CELL DRIVE SURVIVAL
OR EXPANSION BUT SEEMS TO BE
VIRTUALLY DETERMINISTIC OF THE
DECISION TO ENTER LATENCY.
AND COMMONLY USED LRAs DO NOT
ACTIVATE LATENT IN OUR MODEL AND
REAL RESERVOIR CLONES.
DIFFERENCE IS WE CAN NOW GROW
THE CELLS TO HUNDREDS OF
MILLIONS AND DO CHEMISTRY TO
LEARN EXACTLY ABOUT THE
MECHANISMS THAT GOVERN THE
DECISION AND CHANGE THEM TO
ACHIEVE THIS UP HERE.
LET ME CLOSE BY ACKNOWLEDGING
V
VIRENDER PAL AND FRAUK AND MARIE
AND BRAD JONES AND HIS TEAM FOR
THE WORK ON THE AUTHENTIC
RESERVOIR CLONES AND MICHAEL
BETTS AND HIS GROUP DID THE
ANALYSIS.
THANK YOU.
>> GOOD MORNING.
I'M GOING TO FOLLOW UP ON THAT
WITH WHAT WE'RE TRYING TO DO.
SO, WHAT WE'VE BEEN DOING IS
TRYING TO UNDERSTAND THE ROLE OF
ANTIBODIES AS THERAPIES FOR HIV.
AND THE REASON WE'RE DOING THIS
IS ILLUSTRATED ON THE SLIDE IS
THAT ANTIBODIES HAVE TWO ENDS AN
FAB SIDE AND FC SIDE.
AND WHEREAS THE FAB SIDE CAN ACT
LIKE A BLOCKER AND PREVENT
INFECTION.
THE FC SIDE ENABLES THE ANTIBODY
TO HAVE ADDITIONAL FEATURES AND
THOSE ARE ILLUSTRATED ON THE
SLIDE SO FOR EXAMPLE CLEARING
THE VIRUS, KILLING INFECTED
CELLS AND POTENTIALLY MAKING
IMMUNOCOMPLEXES WHICH ENHANCE
POST IMMUNE RESPONSES.
WE'VE DONE THIS WITH TWO
ANTIBODIES THAT WERE CLONED IN
OUR LAB 3BNC117 AND ONE ANOTHER
AND WE'VE DONE QUITE A BIT OF
WORK ON THESE ANTIBODIES AND
HOST IMMUNITY TO HIV.
FIRST IN MICE AND THEN IN
MACAQUES IN COLLABORATION WITH
MALCOLM MARTIN AND HIS
COLLEAGUES AND PLA REFLA CASKEY
HAS TAKEN THIS TO HUMANS.
WE'VE LEARNED A GREAT DEAL ABOUT
ANTIBODIES AND IN PARTICULAR
THEIR SAFETY IN PEOPLE THAT ARE
LIVING WITH HIV.
AND THEIR PROPERTIES IN TERMS OF
THEIR ANTIVIRAL PROPERTIES.
AND JUST TO SUMMARIZE THE
ANTIBODIES ARE SAFE AND SUPPRESS
VIREMIA IN PEOPLE WHO CARRY
VIRUSES THAT ARE SENSITIVE.
MONOCLONAL ANTIBODY THERAPY JUST
LIKE MONOCLONAL DRUG THERAPY IS
ASSOCIATED WITH EMERGENCE
ASSISTANCE AND COMBINATIONS CAN
MAINTAIN VIRAL SUPPRESSION IN
PEOPLE WHO LAR HARBOR VIRUSES
AND JUST LIKE DRUGS IF
ANTIBODIES BECOME
SUBTHERAPEUTIC, PEOPLE LIVING
WITH HIV REBOUND.
AND THE REASON THEY REBOUND, WE
BELIEVE, ALL OF US, IS BECAUSE
THEY CONTAIN THIS RESERVOIR.
SO WHAT IS THIS THING?
TURNED OUT WE'VE LEARNED QUITE A
BIT ABOUT THE RESERVOIR AS A
COMMUNITY OVER THE LAST FIVE OR
SO YEARS.
IN FACT I BELIEVE WE'RE SORT OF
AT AN INFLEXION POINT IN
UNDERSTANDING WHAT THIS IS FIRST
OF ALL UNIQUE CELLS WITH UNIQUE
PROVIRAL INTEGRATIONS BUT
COMPOSED OF CLONES OF CD4 T
CELLS.
THESE CELLS ARE RARE AND HAVE NO
MARKERS TO SPECIFICALLY PULL THE
CELLS OUT.
AND A CONTRACTION OF HALF LIFE
AND THERE'S INTACT VIRUSES AND
DEFECTIVE VIRUSES AND THEY'RE
QUITE DIFFERENT IN THE WAY THEY
BEHAVE WHEREAS THE IMPACT
VIRUSES HAVE A HALF LIFE AND
DEFECTIVE ONES DON'T SEEM TO.
THEY JUST STAY THERE.
AND THIS THING IS DYNAMIC.
THE CLONES EXPAND AND CONTRACT
AND SOME UNDERSTANDING OF THAT
IS RELATED TO THE FACT THAT
THESE T CELLS ARE BEHAVING LIKE
NORMAL CD4 T CELLS AND
RESPONDING TO ANTIGEN.
NOW, IF IN FACT THESE VIRUSES
ARE RESPONDING TO IMMUNOLOGIC
PRESSURE IT MAY BE BECAUSE
THEY'RE BEING EXPRESSED AND SO
ANTIBODIES HAVE THE POTENTIAL OF
RECOGNIZING THESE PROVIRAL
INTEGRATIVE T CELLS.
SO WHAT WE'VE BEEN TRYING TO
UNDERSTAND IS AND WE TAKE
AUTHENTIC INTEGRATION SITES.
THINGS WE, AMY KLINE UP OUR LAB
AND MATIAS IN THEIR LAB AND
REPRODUCE THEM.
FIRST BY CRISPR METHODS AND USE
IT TO INDICATE AN INDICATOR
VIRUS EXACTLY IN THE SAME
POSITION, NUCLEOTIDE FOR
NUCLEOTIDE WHERE WE KNOW
AUTHENTIC LATENT VIRUSES COME
FROM AND THEN ASK HOW DO THEY
ARE BEHAVE.
THIS IS ILLUSTRATED IN THE PAPER
THAT WAS RECENTLY PUBLISHED AND
HERE ARE SIX DIFFERENT
INTEGRATION SITES AUTHENTIC
SITES REPRODUCED -- HERE FOR
EXAMPLE YOU SEE THE PINK SHOWS
IN THIS PARTICULAR INTEGRATION
PROVIRUSES EXPRESS.
IN THIS ONE IT'S NOT.
IN THIS ONE IT'S NOT BUT WHEN WE
ACTIVATE IT YOU CAN SEE THAT IT
IS EXPRESSED.
SO, THESE SIX ARE BEHAVING IN
TWO OR THREE DIFFERENT WAYS.
SOME EXPRESSED AT BASELINE,
OTHERS ARE NOT.
THE LEVEL OF EXPRESSION IS SHOWN
HERE COMPARED TO AN AUTHENTIC IN
VITRO EXPRESSION YU2 AUTHENTIC
VIRUS AND SEE THE LEVEL OF
EXPRESSION IN THE ONES
EXPRESSING IS MUCH LOWER BY LOGS
THAN THE ACTIVE.
WHEN WE TRIED TO LOOK AT THE
RELATIONSHIP BETWEEN THE GENE
AND SITE OF INTEGRATION AND THE
EXPRESSION OF THE PRO VIRUS WHAT
WE SEE IS SOMETHING LINEAR WHICH
IS SIMILAR TO WHAT PAUL WAS
TALKING ABOUT WHICH IS THE
PROVIRUS EXPRESSION IS DICTATED
BY THE GENE AND NOT VICE VERSA.
AND THE VIRUS HERE IS NOT
CHANGING THE EXPRESSION OF THE
GENE.
THE SECOND APPROACH IS TO CLONE
OUT THE ACTUAL CELL THAT CARE
RID THE PROVIRUS.
THE IMPACT PROVIRUS AND THIS IS
THE APPROACH WE'VE TAKEN IN
COLLABORATION WITH BRAD AND
BASED ON OUR OWN WORK ON T CELL
RECEPTOR EXPRESSION IN CLONES OF
CELLS BASED ON T CELL RECEPTOR
WHICH ENRICHED THE POPULATION
THAT WE WANT AND THEN GO AHEAD
AND CLONE IT.
WHAT WE'VE LEARNED BY COMPARING
EX VIVO CELLS.
CELLS STRAIGHT OUT OF THE
INDIVIDUAL AND CELLS THAT WE'VE
GROWN IN CULTURE BY 10X IS
THEY'RE RELATIVELY SIMILAR IN
TERMS OF GENE EXPRESSION.
SO WE HAVEN'T CHANGED THEM VERY
MUCH.
AND THE OTHER THING THAT WE'VE
LEARNED IS HIV IS EXPRESSED IN
THESE CELLS BUT AT VERY FAR
LOWER LEVELS THAN AN AUTHENTIC
INFECTION.
AND AGAIN THE PROVIRAL
INTEGRATION IS NOT CHANGING THE
EXPRESSION OF THE GENE TO WHICH
THEY ARE EXPRESSED.
THIS IS A SMALL NUMBER.
IF ALL OF NA -- THAT IS TRUE IS
THE BASIS OF WHAT WE'RE TRYING
TO UNDERSTAND THE INFUSION OF
CELLS IN TRIALS THEY'RE DOING
IMPACTING THE RESERVOIR AND
WHETHER OR NOT THE INFUSION OF
ANTIBODIES CAN HELP TO
ACCELERATE THE RESERVOIR.
I MENTIONED THE WORK WAS HEADED
BY AND WE HAVE HELP FROM A
NUMBER OF OTHER PEOPLE AND
COLLABORATED TO GROW CELLS IN
VITRO.
>> SO PAUL AND MICHEL HAVE
TALKED ABOUT THE IDEA THAT SOME
CELLS CAN CONTINUE TO BE
RECOGNIZED BY HIV AND RESIST
BEING KILLED.
FOR A LONG TIME WE'VE BEEN
STRUGGLING HOW TO STUDY THE RARE
CELLS AND HAVE A SOLUTION THAT'S
BEEN TALKED ABOUT THROUGH A LOT
OF EFFORT AND LOVE WE CAN GROW
CLONES, TENS OF MILLIONS OF
CELLS FROM HARBORING CELLS FROM
THEM AND STUDY THEM AND THIS
EXPRESSES HIV AND COMES FROM A
PERSON LIVING WITH HIV IN
CONTRAST TO OTHER LATENT CLONES.
HOWEVER, IT'S MASSIVELY EXPANDED
IN THE INDIVIDUAL.
SO WHY IS THIS CLONE PERSISTING?
THE QUESTION IS DOES THE
INFECTED CELL SURVIVE THE ATTACK
COMPARED TO THE UNINFECTED CELL
AND WE HAVE THE CELLS IN THE
SAME WELL TO SERVE AS A NICE
CONTROL.
SO WITH ALL THAT THIS IS WHAT
THE RESULT LOOKS LIKE.
WE'RE VERY EXCITED TO SEE THIS.
SO WHEN YOU ADD CTL THE PEP SIDE
PULSE CELLS ARE KILLED OFF
EFFICIENTLY AND THE ONLY
SURVIVORS YOU SEE ARE THE
INFECTED CLONE.
THERE'S NO SURVIVORS FROM THE
SAME WELL.
THAT'S ONE WELL.
THESE ARE MULT PRAL SUB CLONES
THE OF THE SAME CLONE FROM AN
INDIVIDUAL AND AFTER CTL KILLING
WE HAVE MULTIPLE SURVIVE.
WE CAN NOW STUDY THIS CLONE
SINCE WE HAVE IT ISOLATED.
THIS ONE DOESN'T GROW QUITE AS
WELL AS WE'D LIKE BUT CAN GET
SOME ANSWERS.
THE FIRST THING WE OBSERVED IS
THE OVEREXPRESSED FAMILY MEMBER
BCLXL A PROSURVIVAL MECHANISM
MAY HELP THE CELL SURVIVE AND
WORK FROM OTHERS SHOWED IT CAN
BE OVEREXPRESSED IN CELLS THAT
HARBOR THE HIV RESERVOIR AND
THERE'S CLINICAL TRIALS ONGOING
TO TEST WHETHER BCL2 INHIBITORS
CAN REDUCE THE RESERVOIRS.
WHAT JUMPED OUT TO US FROM THE
SEQUENCING DATA WAS THIS CLONE
IS HIGHLY CYTOTOXIC COMPARED TO
UNINFECTED CELLS CLONED IN
PARALLEL AND EXPRESSED AN NK
MARKER AND AN NK CD4 T CELL.
WE'LL COME BACK TO THAT BUT WHAT
WE WANT TO DO IS HAVE
THERAPEUTIC TARGETS TO OVERCOME
CTL RESISTANCE.
WE CAN GET CLUES FROM PROFILING
THE RESERVOIR CLONE AND NEED TO
KNOW WHAT THEY LOOK LIKE AND
HOPE IT WILL HELP THE FIELD AND
HERE'S HOW WE DEFINE AND
CHARACTERIZE CTL RESISTANCE
CELLS IN GENERAL AND BEGAN WITH
MEMORY CELLS AND INFECT WILL
THEM WITH WILD TYPE HIV OR ONE
POINT HIV IDENTIFY BY A CLONE
AND CHALLENGED AND WE KILL MOST
THE CELLS THAT HAVE WILD TYPE
VIRUS TRARGETED AND DON'T KILL
THE BY STANDARDS AND ARE THE
CELLS THAT SURVIVE THE FIRST HIT
RESISTANT OR LUCKY IN THE FIRST
ROUND?
TO TEST THAT WE COME IN WITH THE
SECOND CLONE THAT TARGETS THE
EPITOPE AND NOW EVERYTHING'S A
TARGET.
THE CELLS NOT CHALLENGED IN THE
FIRST ROUND ARE KILLED.
WHEREAS THE CELLS THAT SURVIVED
AND SELECTED IN THE FIRST ROUND
CONTINUE TO SURVIVE.
SO DIRECT EVIDENCE THE CELLS
HAVE SOME ABILITY TO RESIST CITO
TOXIC T CELL AND WE CAN
CHARACTERIZE WHAT MAKES THE CELL
SPECIAL.
THIS IS BULK RNA SEQ AND THERE'S
A DISTINCT PROFILE AND THE
SINGLE CELL LETS US DIVE IN THE
HETEROGENEITY AND IT DEFINES 11
CLUSTERS.
WHEN WE EXPOSED THE CTL WE SEE
THE SURVIVORS HAVE ENRICHMENTS
IN SOME CLUSTERS AND DEPLETIONS
IN OTHERS.
THERE'S VARIABILITY IN THEIR
SUSCEPTIBILITY TO CTL.
THEY'RE FLOW STARTED BASED ON
THE EXPRESSION BEFORE GOING INTO
THIS.
THEY'RE ALL EXPRESSING HIV.
AS THE NOT ABOUT LATENCY AND NON
LATENCY IN THE EXPERIMENT.
THE SURVIVING CELLS ARE ALL
EXPRESSING HIV.
SO WHAT CAN BE LEARNED ABOUT THE
CLUSTERS.
THE CLUSTER THAT WAS MOST
CONSISTENTLY ENRICHED IS THE
CLUSTER 9 AND IF YOU ASK WHAT
THE TRANSCRIPTOMIC DATA AND WHAT
POPS OUT IS ITS CYTOTOXIC CD4T
CELLS AND THE REAL CELLS IS
RE-ENRICHED IN THESE CELLS AND
USING THE SAME MODEL TO DEFINE
THIS CLUSTER THE CELLS
EXPRESSING RNA IN VIVO ARE
ENRICHED WITH THE CLUSTER OF
CELLS WE ARE STUDYING.
HERE'S THE FIRST THING AT THE
CENTER OF THIS VEN DIAGRAM
CYTOTOXIC CELLS ARE
HETEROGENEOUS AND ASSOCIATED
WITH CELLS THAT RESIST TL
KILLING.
SO WHY ARE CD4 CELLS CYTOTOXIC.
THEIR JOB IS TO KILL OTHER CELLS
AND DON'T WANT TO SELF-DESTRUCT
SO IT'S LIKE A FLAKE BEING
IMMUNE TO IT'S OWN VENOM LIKE A
SNAKE THAT BITES ITS OWN TONGUE.
HOW DO THEY PROTECT THEMSELVES.
SO ONE OF THE MECHANISMS THAT'S
BEEN IN THE FIELD'S
CONSCIOUSNESS HAS BEEN AN
INHIBITOR FOUND TO BE EXPRESSED
IN CD4 T CELLS AND WORK HAS
SHOWN IF YOU KNOCK THIS OUT YOU
DO NOT IMPROVE KILLING OF THE
CELLS.
SO IT SEEMS TO NOT BE THE MAJOR
THING.
HOWEVER, THE RESERVOIR CLONE DES
PLAYS TWO KNOWN MECHANISMS BY
WHICH CYTOTOXIC T CELLS PREVENT
SELF-DESTRUCTION.
IF YOU LOOK AT THE CELLS THAT
RECEIVED A HIT THE UNINFECTED
CLONE DIES AND ESSENTIALLY IT
GETS A HIT AND HITS BACK.
AND IT'S KNOWN TO BE MECHANISM
BY WHICH NATURAL KILLER CELLS
PROTECT THEMSELVES FROM CELLS IN
ENZYME.
AND SOME CELLS RELEASE INTO THE
SYNAPSE TO PROTECT THE CELL.
IF WHERE HE LOOK AT THE
RESERVOIR CELL IT OVEREXPRESSES
AND EXPOSES IT AT THE SURFACE
AND IT GRANULATES.
THE NEXT STEP IS TRYING TO
OVERCOME AND SEE IF WE CAN
IMPROVE KILLING OF THIS CLONE.
I CAN'T TELL YOU JUST ABOUT THE
ONE CLUSTER SO WHAT'S THE MOST
DIFFERENT THING ABOUT ALL THE
CELLS.
SURVIVE ARE METABOLICALLY QUI --
QUIESCENT AND THEY HAVE LESS
REACTIVE OXYGEN SPECIES AND CAN
DO FUNCTIONAL ASSAYS AND SEE THE
SURVIVORS HAVE LESS CAPACITY AS
WELL AS LESS GLUCOSE DEPENDENT
METABOLISM AND SELECTED FOR
CELLS LOW FOR THIS TOXIC
BYPRODUCT AND WE CONSISTENTLY
SEE THAT.
WHY IS METABOLISM IMPORTANT?
WE THINK THIS IS THE REASON.
SO WHEN A CYTOTOXIC T CELL
DELIVERED A HIT TO A TARGET CELL
ENZYMES REACT MITOCHONDRIA AND
THESE DAMAGE DNA IS A BIG PART
OF WHY THE CELL DIES.
IF THE CELL DOESN'T HAVE ENOUGH
OF THESE IT CAN EXPLAIN WHY IT
LIVES.
THE DRUG WE SHOWED IMPROVE LEVEL
OF RE-OXYGEN SPECIES IN CELLS
AND IMPROVES KILLING OF CELLS.
SO HERE WE'RE SEEING MORE THAN
98% KILLING OF HIV INFECTED CD4
CELLS AND I'VE BEEN DOING
KILLING ASSAYS FOR YEARS AND
THIS IS THE MOST POTENT KILLING
I'VE SEEN.
I'M PRETTY EXCITED ABOUT THIS.
THIS IS FROM PUBLISHED WORK AND
THE CELLS THAT TRANSCRIBE IN
VIVO ALIGN IN MANY WAYS WITH THE
METABOLIC QUIESCENT PROFILE OF
OUR CELLS AND MAY BE AFFECTING
HOW THESE SURVIVE.
SO, MARINA MENTIONED WE'RE IN
THE DISCOVERY TO TRANSLATION
PIPELINE.
I DON'T HAVE TIME TO TALK ABOUT
SOME MUCH THESE TODAY AND WANTED
TO TALK ABOUT THE LEADING EDGE
OF OUR WORK SHOWING IN VITRO AND
IN THE LAST 30 SECONDS OR SO I
WANT TO GIVE A NOD TO THE
INHIBITER METASTAT WE THINK IS
POISED FOR CLINICAL TRANSLATION.
THOUGH I'M NOT GIVING MUCH TIME
TO IT TODAY THERE'S A POSTER
HERE AS WELL AND THIS IS A CLOSE
COLLABORATION WITH THE HIV
COLLABORATORY ALSO.
WHAT DOES TESMETASTAT DO?
WE SEE SELECTION FOR CELLS LOW
TO CYTOTOXIC T CELLS AND IT ACT
TO INCREASE MHC1 ON THE SURFACE
OF INFECTED CELLS IMPROVING
KILLING FOR T CELLS IN VITRO.
A FEW SNIPPETS FROM THE MOUSE
MODEL GIVING IT AT THE TIME OF
INITIATION WE SEE LESS RESERVOIR
SEATING AND THE MOST REMARKABLY
WE SEE PROFOUND REDUCTIONS IN PD
1 ON CD 8 T CELLS IN THE MICE
AND THEY'RE SUSTAINED FOR WEEKS
AFTER THE DRUG IS GONE.
WE'RE NOT SURE IF THAT'S ANTIGEN
LOAD OR PROGRAMMING OF THE
CELLS.
SORRY TO DO THAT QUICKLY BUT
PLEASE SEE THE POSTER.
TO WRAP UP I WANT TO ACKNOWLEDGE
THE TERRIFIC COLLABORATIONS WITH
EVERYONE IN REACH AND THE WORK
IS ONLY POSSIBLE BECAUSE OF WORK
WITH THE LABS AND ISABELLA LEADS
THE TEAM DOING THE GREAT WORK
WITH THE CLONES AND OTHERS
LEADING THE HCH2 PROJECT AND
THANK YOU FOR YOUR ATTENTION.
>> WE HAVE ABOUT 15 MINUTES FOR
DISCUSSION.
ANYONE WITH A QUESTION, PLEASE
STEP UP TO THE MICROPHONE AND
SAY WHO YOU ARE AND WHERE YOU'RE
FROM AND LET'S GO AHEAD.
I CAN'T SEE YOU VERY WELL FROM
HERE SO GO AHEAD.
>> HI, MIKE BETTS FROM
UNIVERSITY OF PENNSYLVANIA.
FIRST COMPLEMENTS TO ALL OF YOU.
IT'S FANTASTIC WORK.
MY QUESTION IS FOR PAUL.
IT'S INTERESTING DATA ABOUT THIS
MOUSE MODEL.
I'M CURIOUS AS YOU FLASH BY THE
INTEGRATION SITES AND THINKING
WHAT THE GENES WERE I DIDN'T
NOTICE A LOT OF OFFICE
IMMUNOLOGICAL GENE INTEGRATIONS
WHICH MADE ME WONDER IN YOUR NSG
MOUSE MODEL WHICH IS A CLEAN
FACILITY CAN YOU ALTER A MORE
PHYSIOLOGICAL MODEL TO US WOULD
THAT THEN CHANGE YOUR PROFILES?
>> SO, SINCE WE'RE THINKING OF
PROVIRUSES VERY MUCH A
PASSENGER, ANYTHING THAT WOULD
CHANGE THE PROFILE OF THE CELLS
THAT PROLIFERATE WOULD LIKELY
CHANGE THE SPECTRUM OF CLONES
THAT GROW OUT.
I STILL THOUGH THAT IF
INTEGRATION IN THE GENE HAD SOME
INTRINSIC ABILITY TO DRIVE
PROLIFERATION WOULD COME OUT IN
THE CLEAN ENVIRONMENT.
>> THIS IS FOR PAUL AND JOE.
WHATEVER GETS EXPRESSED ON THE
CELLS IN THE MICE, IS THAT
IMMUNOGENIC IN THE MICE?
>> THEY'RE NFG MICE THEY
SHOULDN'T CARE WITH BINDING
PEPTIDE.
>> OKAY.
>> JIM RILEY, UNIVERSITY OF
PENNSYLVANIA.
I'M SORRY BUT WHY DIDN'T HIV
COME OUT?
LIKE WHAT HIV DID YOU PUT IN?
>> IT DOESN'T HAVE STRUCTURAL
GENES IT HAS REPORTER GENES
INSTEAD.
>> AND HAVE YOU CONSIDERED THE
REASON -- WELL, THAT WAS
COVERED.
THE CULTURE, RIGHT, AFFECTS
WHAT'S GOING ON, RIGHT.
I'M SURE THEY'RE CULTURED MORE
MONTHS GOING FROM ONE CELL TO
BILLIONS.
>> YEAH.
>> SO ARE YOU SELECTING FOR --
IS THERE A PROCESS BY ONES THAT
WOULD COME OUT ARE LIMITED IS
WHAT I'M ASKING.
>> THERE'S NO MECHANISM FOR
ELIMINATION OF CELLS IN THE
MOUSE WE KNOW ABOUT.
>> ARE YOU ASKING ABOUT THE IN
VITRO CLONE CULTURES?
>> KIND OF BOTH, RIGHT?
IN VITRO --
>> FOR THE AUTHENTIC CELLS WE
TRY TO HAVE A LOOK AT THEM WHEN
THEY'RE BRAND NEW IN CULTURE AND
HAD RECENT CLONES WE SAW EXPAND
50 FOLD IN THREE DAYS AND THE
MOST GAG EXPRESSION WE SAW IS
.3%.
SO WE DO CULTURE THEM FOR A LONG
TIME BUT WE HAVE AN OPPORTUNITY
TO LOOK WHEN THEY'RE RELATIVELY
NEW AND AS MICHEL SHOWED THE
TRANSCRIPTIONAL PROFILE IS IN
VIVO.
>> OBVIOUSLY THERE HAS TO BE
CLONES WHERE HIV COMES
IMMEDIATELY OR DO YOU -- OR DID
YOU NOT SURVEY THOSE?
>> THEY DO VARY IN THE CLONES
THAT WE GROW IN VITRO.
THE ONES THAT EXPRESS LET'S SAY
10% IS MAYBE THE HIGHEST WE'VE
SEEN AND THOSE DO TEND TO NOT
STICK AROUND AS LONG AS THE
OTHERS ARE HARDER TO GROW WHERE
ONES THAT EXPRESSION 1% OR 2%
ARE WE MISSING SOME THAT EXPRESS
HIGHER PERCENTAGES?
THAT'S POSSIBLE.
>> REMEMBER TO RECEDE
REPLICATION YOU ONLY NEED ONE
CELL.
1% WILL DO IT.
>> DAVE.
>> DAVE, USC.
THAT WAS BEAUTIFUL AND A LOT OF
DATA SO I'M SURE I MISSED
SOMETHING.
I HAVE A CHICKEN AND THE EGG
QUESTION IN THAT YOU GET TO A
CELL THAT'S METABOLICALLY
INACTIVE OR LESS ACTIVE BEING
RESISTANT.
IS IT THE CELLS OR THE CELL'S
EXPRESSION OF THE VIRUS?
IN YOUR CONTROL WHERE YOU PULSE
WITH PEPTIDE, HOW DID YOU
COMPARE PRESENTATION IN PULSE
CELLS TO THE OTHER CELLS TO SEE
THEY ARE WERE MRAATCHED TO BE YR
CONTROL.
>> IN THE PULSE CELLS IN VITRO
THEY EXPRESSED SIMILAR AMOUNTS
OF MHC1.
I CAN'T SAY WE HAVE A PEPTIDE TO
DIRECTLY QUANTIFY THAT.
WE HAVE LARGE EXPRESSES OF
PEPTIDE, 10 MOLAR AND WE CAN
LOOK AT GAG EXPRESSION IN THE
10X OR SORRY, HIV TRANSCRIPTION
IN THE 10X AND DON'T SEE A
RELATIONSHIP BETWEEN THE AMOUNT
OF HIV TRANSCRIPTION AND THE
METABOLIC STATE WITHIN THIS POOL
OF CELLS THAT ARE EXPRESSING
HIV.
>> BECAUSE THE FOUR QUADRANTS
YOU HAD BOTTOM LEFT THERE WAS A
LITTLE BIT OF SIGNAL AND NO
SIGNAL IN THE BOTTOM RIGHT AND
THE TOP ETCETERA.
SO IS THE PRESENTATION OF TARGET
IN THE PULSE CELLS?
HOW DO YOU NORMALIZE THAT
AGAINST PRESENTATION OF TARGET
IN THE UPPER CELLS.
>> YEAH, SO WHAT I CAN TELL YOU
IS WE DO DEGRANULATION WITH THE
PEPTIDE PULSE INFECTED OR
UNINFECTED CLONE AND IT
DEGRANULATES IN RESPONSE TO B
BOTHPLBOTH
MAXIMALLY RESPONDS AND CAN'T SAY
THERE'S YOU CAN'T TRIGGER THE
CELL.
IT'S A BODY OF WORK I DIDN'T
HAVE A CHANCE TO PRESENT TODAY
BUT THEY'RE LOADED WITH EXCESS
PEPTIDE AND CAUSE DEGRANULATION
IN ISOLATION.
>> I JUST WANTED TO CLARIFY
THIS.
SO ARE YOU SAYING AS THE
RESISTANT CELLS INHERENTLY
EXPRESS LOWER LEVELS OF MHC AND
THAT'S THE BASIC MECHANISM FOR
RESISTANCE TO KILLING?
>> YEAH, THAT IS ONE OF THE
MECHANISMS AND 10X LET'S US
TEASE THESE A PART TO SEE THE
LOW AND HIGH SURVIVORS AND
THERE'S A LOT OF SURVIVORS THAT
ARE MCH1 HIGH SO TRAZ NOT JUST
ABOUT MCH1 HOUR YOU'RE RIGHT,
MHC1 IS AN ISSUE AND WE TREAT TO
WE SEE BETTER KILLING AND THAT
THE ONLY THE CASE WITH WILD TYPE
VIRUS.
WE DON'T SEE AN IMPROVEMENT WITH
THE DELTA F VIRUS BUT SEEMS
DRIVING MHC1 PLAY COUNTER
BALANCE BY DOWN REGULATION.
>> I WAS THINKING IT MAKES A
NICE STORY FOR HOW THE CELLS
TRANSITION TO LATENCY AND ESCAPE
THE IMMUNE KILLING AS A MODEL.
>> RIGHT.
THEY ARE START OFF WITH LESS
ANTIGENICITY.
I DON'T THINK LATENCY AND
SURVIVABILITY ARE MUTUALLY
EXCLUSIVE.
>> THIS IS AN EXCELLENT
PRESENTATION, THANK YOU GUYS FOR
THAT.
LOOKING FORWARD TO HOW WILL IT
BE TRANSLATED TO THE COMMUNITY
MEMBERS WHO WILL HOPEFULLY
RECEIVE THIS AMAZING WORK YOU
GUYS ARE DOING?
>> SO THE WORKSHOP THE IDEA IS
TO BRING IN INDIVIDUALS AND ONE
OF THE REQUIREMENTS IS THAT
THERE'S A TRANSLATION COMPONENT.
SO WE WILL HAVE RESERVOIR SPACE
FOR ONE CAB MEMBER FROM EACH OF
THE COLLABORATORIES
PARTICIPATING AND I THINK
THERE'S STILL ROOM FOR
ADDITIONAL COLLABORATION IF
OTHERS ARE INTERESTED IN
PARTNERING THERE.
BUT THE IDEA IS FOR THOSE WHO
ATTEND WILL GAIN SKILLS IN
SCIENCE TRANSLATION AND BE ABLE
TO COMMUNICATE THAT VIA A BLOG
OR ADDITIONAL SOCIAL MEDIA
COMPONENTS BUT HAVE AN ACTIVE
OUTREACH COMPONENT TO IT.
>> AND WE HAVE HDA2.
DO YOU KNOW IF IT PLAYS A ROLE
ACTING ON THE SAME CELL OR BY
TARGETING THE SPOT YOU CAN
TARGET DIFFERENT CELLS?
>> IT'S A REALLY GREAT QUESTION
AND IT'S SOMETHING THAT WE WANT
TO TEASE OUT WITH OUR 10X DATA
AND HAVE DONE SOME WORK ON THAT.
WHAT I CAN SAY AT THIS POINT IS
BOTH WE NEED TO DO MORE AND IT'S
ALSO NOT CUT AND DRY LIKE YOU
LOOK AT THE U MAP AND HERE'S THE
BCL2 POSITIVE CLUSTER THAT
SURVIVED AND THE MHC1 CLUSTER.
THERE'S MORE AND THE BEST ANSWER
I CAN GIVE FOR NOW IS I THINK
EACH OF THE AGENTS WILL PROBABLY
IMPACT SOME OF THE TARGET CELLS
BUT YOU YOU'LL NEED COMBINATIONS
TO GET A LOT OF THEM.
THERE'S A LOT OF VARIATION IN
THE DIFFERENT CLONES.
THEY ARE ALL HAVE THEIR OWN
PERSONAL AMOUNTS.
-- PERSONALITIES.
>> JOHN, MIKE BETTS AGAIN.
A QUESTION FOR MICHEL.
YOU SHOWED SORT OF THE MODEL
WHAT YOU'RE LOOKING FOR
RECOGNIZING LATENTLY INFECTED
CELLS THAT PLAY EXPRESS BUT YOU
DIDN'T TELL US ABOUT ANY DATA.
ARE YOU ABLE TO SEE SOME
RECOGNITION?
>> THE BEST DATA WE HAVE COMES
FROM MOUSE STUDY THAT WERE
PUBLISHED SOME TIME AGO THAT
ANTIBODIES HELPED CLEAR INFECTED
CELLS AND FROM ONGOING CLINICAL
STUDY WHERE WE SEE ACCELERATED
RESERVOIR CLEARANCE.
YEAH.
>> OKAY.
THIS WILL BE THE LAST QUESTION I
THINK.
GO AHEAD.
>> I MAY HAVE MISSED IT BUT DID
YOU THINK ABOUT THE SPECIFIC
AUTHENTIC RESERVOIR CLONES?
>> I'M SO GLAD YOU ASKED.
WE HAVE ONE AND EVA SPECIFIC AND
IT'S REALLY GOOD AT KILLING EVB
TRANSFORMED LYMPHOMA CELL LINES.
SO IT'S A CELL THAT HAS AN HIV
PROVIRUS AND CAN FUNCTION AND DO
IT JOB AND KILL LYMPHOMA CELLS
WHICH ARE INTERESTED AND THE
ONES WE FOUND AND ISABELLA HAS A
POSTER THAT WILL SHOW SOME OF
THE EVB SPECIFICITY RESULTS.
>> OKAY.
I THINK THAT WILL BE ABOUT IT
FOR THIS PRESENTATION.
I JUST WANT THE TO THANK THE
REACH GROUP.
IT'S A REAL PLEASURE TO BE THEIR
PROGRAM OFFICER.
AND THANKS TO THE AUDIENCE FOR
ALL THEIR REALLY GREAT
QUESTIONS.
SO I TURN IT OVER NOW TO DIANNE
LAWRENCE AND THE DARE GROUP.
>> A QUICK REMINDER IF YOU HAVE
NOT ORDERED LUNCH YOU CAN GO OR
GO TO CHAT TO DROP DOT AND PLACE
YOUR ORDER.
IT'S VERY QUICK.
FEEL FREE TO DO THAT.
I'M DIANNE LAWRENCE IN THE
BRANCH OF THE DIVISION OF AIDS
AND DARE STANDS FOR THE DELANEY
AIDS RESEARCH ENTERPRISE AND
THIS TRAEM HAS BEEN IN PLACE
SINCE THE FIRST ITERATION OF THE
PROGRAM.
SO, THE DARE TEAM IS BASED IN
SAN FRANCISCO LED BY STEVEN
DEEKS, LOUIS PICKER AND SHARON
LEWIN.
>> THANK YOU, DIANE.
I'M GOING TO PRESENT ON BEHALF
OF THE DARE COLLABORATORY.
SINCE THE FIRST EDITION OF THE
GROUP WE'VE BEEN LARGELY
FOCUSSED ON THE CONCEPT OF USING
IMMUNOTHERAPY TO REDUCE
RESERVOIR AND CONTROL WHAT'S
LEFT.
THAT'S BEEN THE MAIN THEME
THROUGH THE THREE OR SO
COLLABORATORIES INCLUDING THE
WORK WE'LL PRESENT TODAY.
AND WE DO THIS LARGELY THROUGH A
CORE GROUP OF CLINICAL
ADVOCATORS, VIROLOGIST AND
IMMUNOLOGISTS MANY OF WHOM HAVE
BEEN WORKING TOGETHER MANY YEARS
NOW AND I DON'T HAVE TIME TO GO
THROUGH THE STRUCTURE BUT WE
HAVE FOUR WORKING GROUPS.
THE FIRST IS LED BY LILLY COHEN
AND FOCUSSING ON THE ACTIVE
RESERVOIR PROLIFERATION.
SECOND IS LOOKING AT THE
MECHANISMS OF POST INVENTION
CONTROL AND POST TREATMENT
CONTROL.
YOU'LL HEAR ABOUT SOME OF THIS
TODAY AND LOUIS PICKER LEADS THE
RF2 TEAM USING THE MONKEY MODEL
TO DEVELOP IMMUNOTHERAPIES AND
OTHERS HAVE BEEN LEADING EFFORTS
ON THE DRUG UPDATES WE HAVE
TODAY.
THE THEME OF TODAY THING IS
GOING TO BE TRANSLATION.
THE WORK WE DO TO TAKE IDEAS
FROM THE LAB THROUGH THE ANIMAL
MODELS INTO THE CLINIC AND BACK.
AND THIS IS SOMETHING THAT WE'VE
BEEN DOING PRETTY CONSISTENTLY
ALL ALONG.
OUR CURRENT SUITE OF APPROACHES
IN TERMS OF REDUCING THE
RESERVOIR ARE LISTED HERE
FOCUSSING ON LRAs AND DRUGS WE
THINK CAN ACCELERATE CELL DEATH.
THIS HAS BEEN THE FOCUS OF WORK
SHARON WILL PRESENT IN THE R3.
I THINK WE MADE PROGRESS AS WE
HEAR ABOUT THINGS WE SHOULD MOVE
FORWARD TO THE CLINIC AND SHOULD
NOT.
AGAIN, SINCE DAY ONE WE'VE BEEN
FOCUSSED ON IMMUNOTHERAPY
FOCUSSING THROUGHOUT THE ENTIRE
LIFE SPAN OF THEIR THERAPEUTIC
VACCINES VERSUS VECTORS AND
WE'LL HEAR UPDATES ON THE LATTER
AND HAVE DONE WORK LOOKING AT
THE ROLE OF ANTIBODIES IN PEOPLE
AND NOW WE'RE DOING WORK IN THE
NON-HUMAN PRIMATES AND
THROUGHOUT THE PROGRAM WORKING
ON DIFFERENT CYTOKINES AND
MIXING AND MATCHING THEM IN THE
MONKEY MODELS AND DO CLINICAL
TRIALS AND SHARON HAS DONE
CLINICAL TRIALS AND MOVE SAMPLE
BACK AND FORM AND THAT'S HOW WE
WORK.
IN DEVELOPING NEW THERAPEUTIC
APPROACHES WE'VE HAD A FAIR
AMOUNT OF APPROACH ON WAYS TO
MEASURE WHAT WE'RE DOING AND
DEVELOP ASSAYS.
I THINK RACHEL WILL PRESENT
EXCITING DATA TO USE T CELL
PROLIFERATION AS A POTENTIAL WAY
TO PREDICT IMMUNOTHERAPY AND
WHAT WILL WORK AND WON'T WORK
AND WE'VE WORKED IN DEVELOPING
DIFFERENT WAYS TO MEASURE THE
INTACT RESERVOIR.
ALL RIGHT, SO TODAY YOU'RE GOING
TO HEAR THREE PRESENTATIONS.
THE FIRST WILL BE BY LEWIS.
WE'LL TALK ABOUT HOW WE TEST
INTERVENTIONS IN THE MONKEY
MODEL AND DOES DEEP DIVE IN THE
MECHANISM AND HOW WE'VE BEEN
USING THAT TO INFORM THE NEXT
STEP MOVING THIS STEP.
AND THEN YOU'LL HEAR BY RACHEL
WHO WILL SUMMARIZE SOME CLINICAL
TRIALS WE'VE DIDN'T DOING AND
EFFORTS WITHIN AND WITHOUT DARE
POST INVENTION CONTROLLERS AND
USING THAT TO DEVELOP POTENTIAL
BIOMARKERS WE CAN USE TO GUIDE
FUTURE DRUG DEVELOPMENT AND
FINALLY SHARON WILL FOCUS ON HER
WORK LOOKING AT DRUGS AND
REVERSE LATENCY AND LYNNE --
LYNDA WILL END WITH AN UPDATE.
>> FIRST I'LL START WITH THE
WORK OF THE GROUP AND BE
REPRESENTATIVE HERE.
OKAY.
SO IN TERMS OF THE COMMUNITY
SLIDE, WHAT IS IT WE'RE TRYING
TO DO?
FIRST, WE'RE TRYING TO
UNDERSTAND HOW THE IMMUNE SYSTEM
PARTICULARLY THE ADAPTIVE T CELL
SYSTEM INTERCEPTS THE ACTIVATING
VIRUS AFTER THE VIRUS IS
DISCONTINUED AND BY
UNDERSTANDING THE PROCESS TO
IMPROVE UPON IT TO BE ABLE TO
HAVE IMMUNOTHERAPEUTICS THAT
MANIPULATE T CELL SYSTEM AND
VACCINE THAT ALLOWS FOR POST
CONTROL.
WE'VE BEEN WORKING ON THIS
SYSTEM
SYSTEMATE -- AND INSETERCEPTED
THE VIRAL REBOUND AND WHETHER
THE THERAPEUTIC VACCINE CAN LEA
OKAY.
SO WE DID EXPERIMENTS A NUMBER
OF YEARS AGO TO ASK WHAT THE
NATURAL T CELL RESPONSE DOES IN
THE POST ART CESSATION REBOUND
SITUATION AND SHOWED PRETTY
CLEARLY THE T CELLS DO
CONTRIBUTE TO PLATEAU LEVEL OF
VIRAL REPLICATION BUT THEY ARE
DO NOT ACTUALLY SHOW ANY EFFECT
ON THE TIME TO REBOUND.
SO THEY APPEAR TO INTERCEPT THE
RESPONSE LATE AND AFFECT A 1.5
LOG REDUCTION IN VIRAL
REPLICATION.
SO THEY'RE EFFECTIVE BUT APPEAR
TO COME TOO LATE TO EFFECT THE
POST REBOUND SITUATION AND OTHER
DATA THAT SUPPORTS THIS AND IF
AND IF YOU LOOK AT THE
PROTECTIVE ALEELS ON TIME TO
REBOUND SET POINT VIRAL LOW WE
SEE A SIGNIFICANT EFFECT ON SET
TIME VIRAL LOAD BUT NO EFFECT ON
TIME TO REBOUND.
AND THEY'RE UNABLE TO INTERCEPT
THE PROCESS PRIOR TO VIRUS.
WE ASKED THE SIMPLE QUESTION.
ON THE TOP IS WHAT WE THINK IS
HAPPENING WITH THE NATURAL T
CELL RESPONSE DURING THE
INFECTION PERIOD AND CAN WE
CHANGE THE DYNAMICS SO IT PEAKS
PRIOR AND CAN IT HAVE AN EFFECT.
AND THERE'S VIRAL EXPRESSION AND
WE HAVE A PRIMING OF DOSING
EVERY THREE WEEKS AND CAME BACK
WITH ANOTHER DOSE AND TIMED IT
PRIOR TO REGARD RELEASE WITH THE
HOPE OF INCREASING THE PEAK
RESPONSES PRIOR TO VIRAL
OUTGROWTH.
IN FACT THE GROUPS WERE VERY
COMPARABLE IN TERMS OF THE
CONTROL GROUP WHICH IS GOING THE
RABIES VIRUS VERSUS THE GAG
EXPRESSION VIRUS.
AND THIS IS 20% AT PEAK IN TERMS
OF GAG RESPONSES.
AND THE ACD8 RESPONSES ARE
CONSIDERABLY HIGHER AND WE SEE
THE SAME INCREASES AND RESPONSES
IN PERIPHERAL LYMPH NODES AND
THE RESPONSE IS GOOD THE VACCINE
IS DOING WHAT WE HOPED.
WE'RE TRYING THE FINAL BOOST SO
THE PEAK RESPONSE OCCURS IN THE
WEEK AFTER AFTER THE REWHERE WE
KNOW THE VIRUS IS GOING TO BEGIN
TO COME OUT AND WE DID A GOOD
JOB COMPARED TO THE NATURAL
RESPONSE WHICH PEAKS LOWER AND
LATER.
AND WE SEE BLUE DOTS ARE HIGHER
THEN THE GREEN DOTS.
THIS IS PRIOR TO AND 18 DAYS
POST RELEASE AND THE REBOUND HAS
BEEN COVERED WITH HIGHER T
CELLS.
HEARS WHAT WE FOUND.
THIS IS THE FIRST TIME WE'VE
EVER SEEN A DELAY OF NINE DAYS
IN VIRAL REBOUND.
THIS MODEL WHICH WAS DEVELOPED
WITH JEFF AND BRANDON IS TIGHT
REBOUNDING KINETICS.
AND THERE'S A SIGNIFICANT DELAY
IN REBOUND.
WE HAD A GOOD EFFECT WITH THIS
VACCINE.
WE HAD A 2 LOG OF PEAK VIRAL
LOAD MAINTAINED FOR SIX WEEKS
OVER TIME AND THE CONTROL GROUP
MERGED.
AND THIS IS A STEP TOWARDS THE
GOAL BUT WE HAVE NOT ACHIEVED
WITH THIS VACCINE ALONE THE
RESULTS WE HOPED FOR.
THIS IS INTERESTING THOUGH AND
IF YOU LOOK AT THE RESPONSES
VERSUS THE VIRAL OUTGROWTH WE
DID ACHIEVE THE GOAL WE WANTED
TO HAVING THE RESPONSES PEAK
BEFORE THE VIRUS CAME OUT
OPPOSED TO THE RESPONSES
FOLLOWING THE VIRUS COMING OUT
IN THE CONTROL GROUP.
THE QUESTION IS HOW DO WE MAKE
THIS BETTER?
WE HAVEN'T YET ACHIEVED THE GOAL
OF THE POTENT VIRUS AND WE'RE
TRYING TO ADD ADJUNCTIVE
THERAPIES TO FIT THE GOAL.
ANTI-PD1 DID NOT HELP IN FACT IT
SEEMED TO HURT THE OUTCOME AND
OTHER PRODUCTS CAN'T HELP AND A
LITTLE BIT NEGATIVE EFFECTS
OPPOSED TO THE GAG VACCINE AND
SHARON WILL PROVIDE OTHER DATA.
THAT'S IT FOR ME.%
>> UM, NICE TO BE HERE TODAY.
SO, IN DARE AND OUR WORK IN OUR
LAB VERY MUCH COMPLIMENTS THE
WORK THAT LOUIS PRESENTED EXCEPT
WE STUDY HUMANS AND WORK WITH
STEVE DEEKS AND MICHAEL PELUSO
AND UNDERSTANDING WHAT DEFINES
EFFECTIVE IMMUNE RESPONSES
AGAINST HIV REBOUND AND CAN WE
ILLICIT THAT AND WHAT IS
RESPONDING IS MOST IMPORTANT AS
LOUIS ALLUDED TO AND I LIKE TO
THINK OF THIS AS A CAT READY TO
POUNCE AND AS WE UNDERSTAND THE
MECHANISMS BETTER WE CAN INFORM
FUTURE ITERATIVE STUDIES FOR
GETTING CLOSER TO A CURE.
AND SO AS ALL OF YOU KNOW, RARE
INDIVIDUALS WHO ARE WITH HIV CAN
CONTROL THE INFECTION EITHER
SPONTANEOUSLY PRIOR IT STARTING
THE ART, VERY RARELY AND IN SOME
CASES AFTER STOPPING ART WITHOUT
ADDITIONAL INTERVENTION BEYOND
ART.
THOSE INDIVIDUALS OF COURSE ARE
QUITE RARE AND OCCUR AT HIGHER
RATES THAN INDIVIDUALS WITH LOW
RESERVOIRS SO PEOPLE TREATED
EARLIER IN INFECTION BUT DON'T
ACHIEVE HIGH LEVELS AND SOME
MECHANISMS MIGHT CONTRIBUTE TO
CONTROL IN THE NATURAL SETTING
THAT MAY CONTRIBUTE TO
INTERVENTION CONTROL.
THE SMALLER RESERVOIR MAY BE
SOMETHING ESTABLISHED AT THE
TIME OF INFECTION AND MAYBE
INTERVENE ON AND CONTRIBUTE TO
POST INTERVENTION CONTROL
INCLUDING INNATE AND ADAPTIVE
MECHANISMED AND OUR LAB AND
OTHERS HAVE BEEN FOCUSSED ON
UNDERSTANDING THE ROLE OF CD 8 T
CELLS IN PARTICULAR PROLIFE
PROLIFERATIVE CAPACITY TO
RESPOND TO THE VIRUS.
OUR LAB IS FOCUSSED ON THE TCF1
THAT PROMOTES THIS CAPACITY AND
HAVE TWO POSTERS PRESENTING ON
NHP STUDY AND CHARLIE IS
PRESENTING A STUDY FOCUSSED ON
THE PROLIFERATIVE CAPACITY.
SO HOW DO WE PROMOTE THIS OR
HAVE AN IMMUNE RESPONSE FOR
CONTROL AND HAVE THERAPEUTIC
VACCINE STUDIES.
THE NHP STUDY IS THE MOST
STRIKING TO SHOW THAT AND THE
STUDY SHOWED AN INTERESTING
SIGNAL ABOUT THE POSSIBILITY
THEY COULD ELICIT RESPONSES THAT
CAN CONTRIBUTE TO BETTER CONTROL
AFTER TREATMENT INTERRUPTION.
WE'VE PUT BEEN INTRIGUED BY THE
INDIVIDUALS WHO HAVE BEEN SHOWN
TO INDUCE SOME CONTROL TO HAVE
SOME INDUCE CONTROL AFTER
RECEIVING BROADLY RECEIVING
ANTIGEN ANTIBODYING DURING HIGH
LEVEL OF ANTIGEN AT THE TIME OF
ART OR AFTER ART IS STOPPED IN
SEVERAL STUDIES.
AND STILL LIKE LOW NUMBERS OF
INDIVIDUALS RELATIVELY LOW RATES
OF POTENTIAL CONTROL AND NOT A
LOT OF UNDERSTANDING ABOUT WHAT
THE MECHANISMS ARE.
THE STUDY I'LL TALK ABOUT I
THINK MANY ARE FAMILIAR WITH IS
COMBINATION IMMUNOTHERAPY WAS IN
THREE STAGES RAN IN SAN
FRANCISCO.
THEY WERE THERAPEUTIC VACCINE
STUDY AND DELIVERED THERAPEUTIC
VACCINE REGIMENT FOR CONSERVED
ELEMENTS OF HIV WITHIN GAG.
A SECOND STAGE IN WHICH THERE
WAS INTENDED RESERVOIR REDUCTION
AN ADMINISTRATION OF TWO BROADLY
NEUTRALIZING ANTIBODIES ON ART
AND AT THE TIME OF ART
INTERRUPTION THE TWO WERE GIVEN
AND THE HOPE WAS THAT WE WOULD
SEE CONTROL OF REBOUND.
AND WE PRESENTED THE CLINICAL
RESULTS AND THINK IT'S STRIKING
THAT THERE'S NO PLACEBO ARM BUT
HAVE 10 INDIVIDUALS WITH HIV ON
ART GIVEN THE REGIMENT AND THREE
REBOUNDED NORMALLY BUT SEVEN
PEOPLE HAD UNUSUAL LEVEL OF POST
INTERVENTION CONTROL.
ONE PERSON DOESN'T REBOUND AND
SIX PEOPLE FELL INTO THE LOW
LEVEL OF SET POINT.
YOU LOOK AT THE ACCESS IS WEEKS
FROM THE BEGINNING OF REBOUND
AND THE TOP TWO YOU HAVE TWO
PEOPLE WHO WENT OUT SLOWLY IN
THE VIRAL LOADS OVER MANY WEEKS
AND FOUR PEOPLE WHO WENT UP TO
1,000 COPIES PER MILL AND
MAINTAIN WHICH IS A RATE OF POST
ART CONTROL THAT IS ABOVE A
BACKGROUND RATE OF POST
TREATMENT CONTROL SEEN IN
SIMILAR INDIVIDUALS.
SORRY THIS IS BLURRY.
ONE THING WE THINK IS IMPORTANT
OF THE POST INTERVENTION
CONTROLLERS SO THESE ARE THE
INDIVIDUALS IN BLUE AND GRAY ON
HERE.
WHEN WE COMPARED THE SET POINT
VIRAL LOAD SO THIS IS THE
AVERAGE VIRAL LOAD AFTER THEIR
PEAK, AFTER COMING OFF ART IT
WAS SIGNIFICANTLY LOWER THAN THE
VIRAL LOADS THAT WERE MEASURED
PRIOR TO STARTING ART.
WE THINK THIS IS IMPORTANT WE
SHIFT THE NEEDLE ON WHAT THEIR
SET POINT IS AND THIS WAS NOT
DIFFERENT IN THE INDIVIDUALS WHO
WERE NON CONTROLLERS.
SO REALLY THE QUESTION IS WHAT
CAUSED THE POST-ART SET POINTS
TO BE LOWER?
WHAT WAS THE OUTPUT FROM THE
INTERVENTIONS?
I DON'T HAVE ALL THE ANSWERS
TODAY.
WE ARE DOING WILL A LOT OF
THINGS.
WE'RE TRYING TO PUT TOGETHER THE
ANTIBODY PK WITH THE RESERVOIR
MEASUREMENTS WITH A HOST OF
IMMUNOLOGIC MEASURES TO TRY TO
UNDERSTAND THIS.
BUT THE DATA I'LL PRESENT
FOCUSES ON SEVERAL TIME POINTS.
WE WOULD WANT TO KNOW ARE THERE
DIFFERENCES AT BASELINE BETWEEN
CONTROLLERS AND NON CONTROLLERS
THROUGH THE VACCINE PHASE AND
THEN THE MAJOR TIME POINTS WE'RE
LOOKING AT ARE RIGHT BEFORE
TREATMENT INTERRUPTION AND THEN
AS THEY ARE B NAPS ARE WARN OFF
BEFORE KEY TIME POINT AND RIGHT
INTO REBOUND.
AS YOU PRESSURE TEST THE IMMUNE
SYSTEM YOU HAVE TO DO SOMETHING.
WHAT IS IT DOING AND HOW DOES IT
DIFFER BETWEEN PEOPLE
CONTROLLING AND NOT AND HAVE A
SAMPLING TIME POINT FOR
INDIVIDUALS AT THE RELATIVELY
LOW VIRAL LOADS WITHIN THE FIRST
MONTH OF REBOUND WHERE WE WERE
ABLE TO ASSESS HOW THE IMMUNE
SYSTEM WAS RESPONDING AND
THERE'S A LOT OF DATA HERE I
DON'T HAVE TIME TO SHOW BUT
HAPPY TO GO THROUGH WITH ANYBODY
AND WE SHOWED THE B NAB, PK AS
EXPECTED CORRELATED WITH TIME
AND NOT THE PORT-ART SET POINT
SO BNAB WASN'T RELATED TO THOSE
WITH LOWER SET POINTS AND THE
RESERVOIR LEVEL SHOWED MOST
INDIVIDUALS HAVE RELATIVELY LOW
RESERVOIR LEVEL AND FROM
PERIPHERAL BLOOD CD4 T CELLS THE
NON-REBOUNDER HAD LOW LEVELS OF
RESERVOIR AND HAPPY TO COMMENT
MORE BUT NO CORRELATION WITH SET
POINTS AND THE VACCINES
FANTASTIC VACCINE REGIMENT
ELICITED T CELL RESPONSES
AGAINST THE REGIONS.
THE MAGNITUDE IS MEASURE BY ICS
DIDN'T CORRELATE WITH THE
OUTCOMES.
WE'VE GOTTEN A LOT OF MILEAGE
OUT OF USING MASS CYTOMETRY AND
LOOKED AT CELLS THAT RELATE TO
OUTCOMES.
OUR PANELS ARE DESIGN TO LOOK AT
INNATE AND ADAPTIVE IMMUNE CELLS
AND THIS IS NOT ANTIGEN SPECIFIC
CELLS AT THIS LEVEL BUT IS
IMPORTANT FOR CAPTURING THE
CELLS.
SO, ONE THING I WANTED TO SAY IS
WHEN WE USE THE PANELS AND
SORRY, I DIDN'T REALIZE THIS WAS
SO BLURRY BUT WE LOOKED -- THIS
IS LOOKING THE HEAT PERHAPS ARE
MEANT TO REPRESENT WHAT HAPPENS
ACROSS THE COURSE OF THE TRIAL
AND BASELINE AND POST REBOUND 1
AND WE'RE LOOKING AT POPULATIONS
ACTIVATED IN INNATE AND ADAPTIVE
IMMUNE CELLS AND WHAT'S
HAPPENING IN THE CONTROLLERS WE
DECIDED TO LOOK AT AND SEE THE
POPULATIONS GO UP IN FREQUENCY.
NOT SURPRISING AT THE TIME POINT
WHEN THEY'RE RESPONDING TO
VIRUS.
YOU SEE A LOT OF THEM GOING UP
AT THE PRE-REBOUND TIME POINT
AND SEE THEY'RE ALREADY STARTING
TO COME UP BEFORE PEOPLE HAVE
STARTED TO REBOUND.
THIS HAPPENS TO BE TRUE IN BOTH
THE CONTROLLERS IN BLUE AND NON
CONTROLLERS IN RED.
WHAT'S THE DIFFERENCE BETWEEN
THE CONTROLLERS AND
NON-CONTROLLERS.
LOOKING THROUGH HUNDREDS ME AND
MY GRADUATE STUDENT, DEMI,
LOOKING THROUGH HUNDREDS OF
IMMUNE FEATURES AT A TIME.
THE ABSOLUTELY MOST STRIKING
THING I LOVE CD 8 T CELLS BUT
ABOUT WHAT IS STRIKING THE
CONTROLLERS WHEN THEY HIT THE
POST REBOUND 1 TIME POINT HAVE
IN INCREASE IN THEIR CYCLING OF
THE POSITIVE CD 8 CELLS STRIKING
COMPARED TO THE NON-CONTROLLERS
WHO WERE AT A HIGHER VIRAL LOAD
AND ON THEIR WAY TO START LOSING
CONTROL AND COULDN'T CATCH UP IS
OUR INTERPRETATION OF THIS.
THERE'S A NICE NEGATIVE
CORRELATION AND THE INDIVIDUALS
WHO HAVE HIGHER CYCLING NON-CD 8
T CELLS ARE THE ONE THAT GO ON
TO HAVE LOWER SET POINT AND
SUSTAIN MORE SET POINTS OVER
MANY MONTHS.
WE THINK IT'S REALLY INTERESTING
WITHIN THESE CYCLING CD 8 T
CELLS THE INDIVIDUALS WHO HAVE
INTERVENTION CONTROL HAVE A HIGH
CYCLING THAT EXPRESS THE
IMPORTANT TCF1 IMPORTANT FOR
MAINTAINING PROLIFERATIVE
CAPACITY.
AND AGAIN THERE'S A CORRELATION
WITH THE VIRAL LOADS AND IT'S
INTERESTING THE OTHER STRIKING
POPULATION WE SEE IS NOT AN
INCREASE OR UNIQUE INCREASE IN
PLA
PLASMA BLASTS WHICH PUMP OUT
ANTIBODIES AND WE UNIQUELY SEE
THE PLASMA BLASTS FOR MAKING AN
ANTIVIRAL RESPONSE GOING UP IN
IN THE CONTROLLERS COMPARED TO
THE NON CONTROLLERS AND IN THE B
CELL AND T CELL WORK WE'RE
TRYING TO UNDERSTAND HIV
SPECIFICITY AND INTEGRATING THE
RESPONSES WITH OTHER THINGS.
SO WE HAVE MORE DATA.
HAPPY TO CHAT MORE ABOUT THAT
BUT THE CLINICAL OUTCOMES IT'S
AN INTERESTING CLINICAL TRIAL WE
THINK THE COMBINATION ALTERED
NATURAL HIV REBOUND AND PEOPLE
WITH BETTER CONTROLS HAVE IN A
PRESSURE TEST SETTING A BETTER
ABILITY TO POUNCE ON THE VIRUS.
THERE MAY BE A DISTINCT
MECHANISM OF CONTROL RELATE THE
TO RESERVOIR INDIVIDUALS AND WE
HAVE COLLABORATORS INVOLVED FROM
BEGINNING TO NOW AND ONGOING BUT
WANTED TO SHOW YOU MEMBERS OF MY
LAB AND SOME OF OUR
COLLABORATORS INVOLVED IN THE
WORK I PRESENTED.
THANK YOU.
>> THANKS VERY MUCH.
OUR LAB IS LARGELY FOCUSSED ON
APOPTOTIC DRUGS BUT TODAY I'M
GOING TO TALK TO YOU ABOUT OUR
FINDINGS ON THE IMPACT OF
MONOVMO
MONOVALENT REVERSAL AND CD 8 T
CELL FUNCTION.
WE WERE INTERESTED IN THE EFFECT
OF CANCER DRUGS AS MANY
LABORATORIES ARE AND THEIR
EFFECT ON THE RESERVOIR AND
WHETHER THEY ARE EFFECT THE
VIRUS AND HAVE AN EFFECT ON THE
IMMUNE RESPONSE?
THEY ARE CAN REVERSE HIV LATENCY
BUT HAVE LIMITED POTENCY IN
USING PRIMARY CD4 T CELLS.
AND FOUND THE CELL KILLED IN
VITRO AND IN ANIMAL MODEL
RESULTING IN HIGHER VIRAL
REBOUND AFTER NATI.
SO WHAT ARE THE IMPLICATIONS?
HAVE ADVERSE EFFECTS AND FEEL IT
SHOULD BE USED WITH CAUTION IN
REDUCING CONTROL PURE STRATEGIES
THAT INVOLVE CD 8 T CELLS AND
MORE POTENT LESS TOXIC HIV
SPECIFIC AGENTS ARE STILL NEEDED
TO REVERSE LATENCY.
AND WE'VE BEEN PARTICULARLY
INTERESTED IN THESE TWO
COMPOUNDS THAT WERE GIVEN TO US
BY GDCO152 AND 0197 AND
INTERESTED IN THE MONOVALENT
BECAUSE THEY'VE BEEN SHOWN TO
HAVE REDUCED TOXICITY BUT IT'S
WELL KNOWN THE BIVALENCE HAVE
BETTER POTENCY.
THO
HAVE BEEN INVESTIGATED BY
OTHERS SOME IN THE ROOM TODAY.
WE STARTED LOOKING AT THE THE
MONOVALENT AND VALENTS RAND THE
CELL LINE UNDER THE CONTROL OF
AN HIV LTR.
AND AS DESCRIBED EARLIER BY
OTHERS CLEARLY THE BIVALENCE AND
IT'S 10% OF THE CELLS AND THE
HIGHER CONCENTRATIONS THERE'S
CLEARLY EVIDENCE OF CITO
TOXICITY SHOWN HERE.
AND THEY WERE TREATED EX VIVO
WITH AND WITHOUT T AND L ALPHA
AND I WON'T SHOW THAT DATA
TODAY.
AND WE USED THE HIV
TRANSCRIPTIONAL PROFILING ASSAY
INITIALLY DESCRIBED AND RECENTLY
ADAPTED BY MY GROUP TO USE IN
THE SETTING OF DIGITAL PCR
OPPOSED TO DROPLET PCR FOR
HIGHER THROUGHPUT.
INTERESTING IN THE PROFILING
ASSAY BECAUSE IT TELLS US ALL
STAGES OF TRANSCRIPTION
INITIATION AND ELONGATION AND
COMPLETION AND MOST IMPORTANTLY
SPLICING.
ONLY SPLICED RNA ALLOWS FOR
PROTEIN PRODUCTION AND AMPLIFIED
INCOMPLETE TRANSCRIPTS.
WHAT DID WE FIND?
HERE'S THE DATA FOR
TRANSCRIPTIONAL INITIATION.
BLACK SHOWING MONOVALENCE AND
BIVALENCE AND SEE THE GDC
COMPOUNDS 0197 AND SHOWED
INCREASE IN TRANSCRIPTIONAL
INITIATION.
WHEN WE LOOK AT POLY A
COMPLETION THE 8852 GOES ON TO
SIGNIFICANTLY COMPLETE
TRANSCRIPTS WHILE THE MONOVALENT
COMPOUNDS SHOWED NO SIGNIFICANT
INCREASE.
WHEN WE LOOKED AT SPLICING THERE
WAS NO SIGNIFICANT INCREASE IN
SPLICED RNA WHEN TREATED WITH
THE MONOVALENT OR AZD5582.
MONOVALENT AND BIVALENT DOES NOT
INDUCE SPLICING.
WHY DO WE SEE BLIPS IN MACAQUES
IN THOSE TREATED WITH A ZD5582
AND COULD THIS BE EXPLAINED WITH
THE IMMUNOLOGICAL EFFECTS OF
SMAC AND CAN THEY PLAY IN CURE
STRATEGIES?
THERE'S DATA IN THE REGULATORY
EFFECTS OF SMAC MIMETICS AND
INHIBITORS OF ATPOPTOSIS PROTEIS
WERE REQUIRED FOR EXPANSION AND
SURVIVAL DURING ANTIVIRAL
IMMUNITY IN MOUSE SO FAVORABLE
EFFECTS OF THE SMAC MIMETIC AND
SHOWED TO ENHANCE SURVIVAL OF
ANTI-TUMOR T CELLS AND THE
BLOCKADE TO PROMOTE TUMOR
IMMUNITY.
WE'RE INTERESTED REALLY IN THE
FAVORABLE EFFECTS OF SMAC MIM
ETIET
IB
ICES.
WE TOOK PEOPLE ON ANTIRETROVIRAL
THERAPY AND ADDED HIV PEPTIDE TO
INCREASE THE CELLS AND QUANTIFY
THE CELLS THAT HAVE UNDER GONE
PROLIFERATION.
WE THEN INCUBATED THOSE IN THE
PRESENCE OF SMAC MIMETICS AND
LOOKED AT PROLIFERATION AND THE
TOTAL CELLS AND HIV SPECIFIC
CELLS LABELLED WITH TETRAMERS
AND COULD QUANTIFY HIV SPECIFIC
AND TOTAL CD 8 T CELLS.
SO THIS GRAPH SHOWS YOU AND
POSITIVE CONTROL A DRUG
INTERESTED IN IN ITS ENHANCING
AFFECTS OF HIV CD T CELLS.
IT'S SHOWN IN GRAY AND THE DATA
IS RELATIVE AND PLOTTED AT 1.
AND YOU CAN SEE ABOVE THE LINE
ALL THE MONOVALENCE SMAC
MIMETICS SHOW AFTER PROLONGED
CULTURE IN VITRO AFTER DAY 7 YOU
SEE THE PROGRESSIVE INCREASE IN
HIV SPECIFIC T CELLS WHICH FOR
SOME MONOVALENCE T CELLS 0917 IS
QUITE SIGNIFICANT.
IN CONTRAST YOU SEE THE RED LINE
IN THE PRESENCE WHERE THERE'S IN
FACT NO EXPANSION OF THE HIV
SPECIFIC T CELLS AND CELL LOSS.
THESE FINDINGS WERE SIMILAR BUT
LESS DRAMATIC IN CD 8 T CELLS.
SHOWN HERE IN A DIFFERENT WAY
YOU SEE THE LOSS OF LOSS
SPECIFIC CD 8 T CELLS WAS PRETTY
DRAMATIC AND MORE PRONOUNCED
THAN WHAT YOU WOULD SEE WITH THE
LOSS OF JUST TOTAL CD 8 T CELLS.
WE THINK THIS IS A CLEARLY
ADVERSE EFFECT OF AZD5582 AND
EFFECT ON CD4 AND CD 8 T CELLS I
WON'T GO INTO NOW.
AT THE SAME TIME WE HAD THE
OPPORTUNITY TO ASK THE QUESTION
IN COLLABORATION WITH LOUIS AND
HIS TEAM BECAUSE AS SHOWED
EARLIER WERE LOOKING AT THE
EFFECTS OF mRNA IN THE PRESENCE
OF SIV.
THEY COMPARED AN mRNA VACCINE
THAT EXPRESSED GAG AND THE
FAVORABLE AFFECTS THE
VACCINATION HAD ON HIV SPECIFIC
T CELLS AND TIME TO REBOUND AND
VIRAL SET POINT.
AT THE SAME TIME THE TEAM DID
THE IDENTICAL STUDY BUT ALSO
INCLUDED MULTIPLE DOSES OF
AZD5582.
THE INITIAL HYPOTHESIS WAS AZD
WOULD ENHANCE THE VACCINE AND
MAKE THEM MORE VISIBLE TO THE
NEWLY GENERATED CD 8 T CELLS.
SO WHAT DID THEY FIND?
WELL, AS YOU'D EXPECT AZD DID
INDEED INDUCE BLIPS YOU SEE IN
BOTH ARMS OF THE STUDY WITH THE
MONKEYS RECEIVED AN SIV GAG
VACCINE IN THE PRESENCE THEY ARE
WERE BLIPS AND MORE TIME POINTS
WHERE THE VIRAL LOAD WAS GREATER
THAN THE AMOUNT OF DETECTION.
WHAT HAPPENED TO THE HIV
SPECIFIC T CELL RESPONSE?
I'M SHOWING THE mRNA VACCINE
DATA WITH AZD IN THE SOLID AND
WITHOUT IN THE DASH LINE.
FROM LEFT TO RIGHT YOU SEE THE
GAG SPECIFIC CD 8 T CELLS IN THE
PRESENCE OF AZD.
THEY WERE REDUCED GAG SPECIFIC
CD 8 T CELLS.
IN THE PRESENCE OF AZD THE
BENEFICIAL EFFECT OF THE mRNA
VACCINE WAS AB ROW GRW -- ABROG
AND THERE WAS A HIGHER PEAK
VIRAL LOAD AND VIRAL SET POINT.
THESE FINDINGS WERE THE SAME IN
THE CONTROL ARM MEANING WHEN WE
PREPARED THE CONTROL ARM WITHOUT
AZD IN THE DASH LINE AND THE
SOLID LINE YOU CAN SEE LOWER GAG
SPECIFIC CD 8 T CELLS.
NO DIFFERENCE IN TIME TO VIRAL
REBOUND BUT THIS IS REALLY
INTERESTING THAT IN THE PRESENCE
OF AZD YOU ACTUALLY HAD A HIGHER
SET POINT THAN WITHOUT AZD.
SO, IN SUMMARY WE'VE SHOWN
MIMETICS REVERSE LATENCY WIN
PEOPLE WITH HIV IN ART AND EX
VIVO MIMETICS CAN ENHANCE
SPECIFIC T CELLS WHICH WE THINK
IS A FAVORABLE FEATURE AND THAT
WAS SIGNIFICANT TOXICITY.
WE DON'T KNOW IF THIS IS A CLASS
EFFECT OF ALL BIVALENCE AND
WE'RE INVESTIGATING WITH THE
PANEL AND ELIMINATED ANY
EXPANSION OF CD 8 T CELLS AND
UNFAVORABLE OUTCOME.
THE ADDITIONAL OF AZD ABROGATED
IN A BENEFICIAL EFFECT OF THE
VACCINE AND RESULTS IN A HIGHER
VIRAL LOAD SET POINT FOLLOWING
ATI WITH OR WITHOUT THE VACCINE
AND THOUGH NOT DIRECTLY SAVED AN
IN THE STUDY ONE POSSIBLE
EXPLANATION FOR PRODUCING VIRAL
BLIPS IN NON-HUMAN PRIMATE
MODELS COULD BE A RESULT OF
DEPLETING HIV SPECIFIC T CELLS
RATHER THAN DIRECTLY ACTING ON
THE LATENT INFECTED CELLS.
WE THINK MORE POTENT, LESS TOXIC
LRAs ARE STILL NEEDED AND WE'VE
BECOME VERY INTERESTED IN THE
ROLE OF mRNA AND DREW YOUR
ATTENTION AND LOOK AT THE POSTER
PRESENTED BY MICHAEL MOSO FOR
HIV LATENCY REVERSION USING THE
PARTICLES.
I'D LIKE TO ACKNOWLEDGE ALL THE
PEOPLE INVOLVED IN THE WORK AND
A FANTASTIC PARTNERSHIP WITH
OHSU AND JAKE, LEWIS AND THE GDC
DRUGS FROM GENENTECH.
THANK YOU VERY MUCH.
>> HI, EVERYBODY.
GOOD MORNING.
SO, LET'S GO TODAY I'M GOING TO
TALK PRIMARILY ABOUT OUR
COMMUNITY ENGAGEMENT ACTIVITIES
IN A LONG-TERM SURVIVORS'
RESEARCH PROJECT BUT BEFORE I
GET STARTED, MICHAEL AND DEREK
WOULD YOU STAND UP, PLEASE.
WE COULDN'T DO ANY OF THIS
WITHOUT OUR MEMBERS.
I THINK MOST KNOW MICHAEL.
AND THERE'S OUR NEW CO-CHAIR.
I DON'T WANT TO FORGET THEM
BECAUSE THEY'RE INSTRUMENTAL.
AT THIS POINT WE HAVE HALF OUR
PEOPLE ON OUR CAB ARE PEOPLE OF
COLOR AND MANY PEOPLE -- NAURD
--
[NO AUDIO]
WE ALSO INCLUDED LIFE TIME
SURVIVE.
WE DID THAT LAST YEAR.
THIS YEAR WE DECIDED TO TALK
ABOUT THE RESERVOIRS.
AND EVERY YEAR WE'VE LEARNED
SOMETHING FROM THIS.
I THINK WE EXTEND THE SESSIONS
AND MADE THEM NOT JUST
INVESTIGATORS.
WE INCLUDED AT LEAST TWO
COMMUNITY PEOPLE BESIDES A CHAT
BEFORE WE CAN HAVE LIVE
INTERACTION WITH PEOPLE AND THAT
REALLY HELPS.
WE'VE INITIALLY STARTED WITH TWO
NEW INVESTIGATORS AND SO WE'RE
TRYING TO GET THEM ACCLIMATED
LIKE WE ARE THE REST OF THE
COMMUNITY.
AND YOU CAN SEE WHAT WE'VE DONE.
SO, WHAT I'M GOING TO DO INSTEAD
OF STALK -- TALKING ABOUT THE
DIFFERENT TOPICS IS WHAT WE DO.
[NO AUDIO]
AND WE DECIDED TO PRESENT AT THE
SESSIONS.
WE'VE GOT THIS FORMULA WHERE WE
ALWAYS INCLUDE ABLE WHICH MEANS
LIFE TIME AND LONG-TERM SURVIVE
AND ALWAYS INCLUDE WOMEN.
WE DO AN INTRODUCTION INITIALLY.
WE HAD A GREAT SESSION WITH A
NEW RESERVOIR RESEARCH BY
MATIAS.
WE LOVED THAT AND WE HAD A GREAT
SESSION IN NOVEMBER EARLIER THIS
MONTH WITH THE TWO CAB LEADS AT
HOPE AND RUBIN IS ALSO A NEW
INVESTIGATOR.
SO WE'RE TRYING TO INCLUDE
EVERYBODY IN OUR EDUCATION
PROCESS, INVESTIGATORS AND
COMMUNITY PEOPLE ALIKE.
AND IN DECEMBER WE'RE GOING TO
HAVE A BEHAVIORAL STUDY SESSION.
FLAUD
[NO AUDIO]
[NO AU
[NO AUDIO]
BE A BIG SESSION BECAUSE WE
ALSO HAVE ONE OF THE LEADS
RESEARCH PEOPLE AT THE NATIONAL
MINORITY AIDS COUNCIL AND WE
HAVE PAUL A COMMUNITY PARTNER
AND AIDS BALTIMORE IS ALSO A
COMMUNITY PARTNER.
HERE'S ANALYTICS.
SO FAR WE'VE HAD 224 PEOPLE ON
THE CALLED, YOUTUBE AND FACEBOOK
HITS AND AT 70% HIGHLY SATISFIED
AND THAT GETS BETTER EVERY TIME
WE DO THIS BECAUSE AS I SAY
WE'VE LEARNED FROM EACH ONE OF
THESE.
SO WE'RE GOING TO TAKE THE LEAD
ON THE LAST GIFT STUDY UPDATE
AND
[NO AUDIO]
ONE THING HE SAYS IS WE NEED TO
COLLABORATE WITH OTHER GROUPS
AND OTHER PEOPLE AND THAT'S WHAT
WE DO.
IT'S EASIER I THINK WE HAVE A
LEG UP NOT ONLY END WITH THE
FIRST ROUND OF SITES BUT MOST OF
US IN THE GROUP HAVE KNOWN EACH
OTHER 20, 25 YEARS.
WE'RE VERY USED TO WORKING
TOGETHER WITH EVERYTHING FROM
THE BAD OLD DAYS TO WHAT IS NOW
TURNING INTO THE NEW BAD OLD
DAYS.
HOPEFULLY I'M WRONG BUT I'M
USUALLY NOT WRONG ABOUT THAT
STUFF.
THIS IS HOW WE GET NEW PEOPLE IN
FROM DIFFERENT GROUP AND
DIFFERENT COLLABORATORIES AND
IT'S REALLY BEEN VERY VERY
REWARDING.
THIS IS CARRINE'S STUDY.
[NO AUDIO]
INTERESTINGLY 32 PEOPLE WERE
RECRUITED.
69 YEARS OLD WAS THE MEDIAN AGE
AND WHAT WE ARE TRYING TO DO IS
MAKE THAT 65 CUT AND PASTE
NUMBER HIGHER.
IF PEOPLE ARE OTHERWISE HEALTHY
AND NOT PRECLUDED BY THE
INCLUSION OR EXCLUSION CRITERIA
THEY SHOULD BE INCLUDED IN THE
TRIALS IT'S A POLICY THING WE
ARE PUSHING FOR THE POPULATION.
WE HAVE 50% MEN NOT A SURPRISED
AND 47% CISGENDER WOMAN AND
TRANS WOMAN AND 30% OF THE
PEOPLE WERE AFRICAN AMERICAN OR
BLACK.
THE THING IS WE KNOW THIS IS
HARD.
IT WAS HARD FOR US BUT WE'RE
SHOWING YOU THAT IT CAN BE DONE
AND IT'S NOT A HUGE STUDY BUT
YOU KNOW NONE OF OUR STUDIES ARE
REALLY HUGE AT THIS POINT.
I MEAN THIS IS THE WAY TO DO IT
AND HOW WE DID IT WAS I
ALWAYS -- I'M GOOD AT MAKING THE
TRAINS RUN.
I GOT MONEY FROM EACH ONE OF
THESE INSTITUTIONS FROM CRISPR,
DARE AND GRID.
WE MADE A CONTRACT WITH U.C. SAN
DIEGO AND DID THE STUDY.
IT WAS INTERESTING.
IT WASN'T REALLY HARD BECAUSE
INVESTIGATORS WERE BEHIND US AND
HELPED GET THE MONEY OUT THE
DOOR.
AND EVERY TIME WE'VE DONE A
PRESENTATION IN FRONT OF PEOPLE
THAT ARE LONG-TERM SURVIVORS OR
EVEN PEOPLE THAT ARE NOT, OKAY,
I'M GOING TO BE CURED.
MY SUPPORT GROUPS ARE GOING TO
GO AWAY.
MY DRUG COVERAGE IS GOING TO GO
AWAY.
I WON'T HAVE MEDICAL CARE.
WE REALLY NEED TO THINK ABOUT IF
WE'RE SUCCESSFUL WHAT ARE THE
RAMIFICATIONS OF THAT AND I
THINK THAT'S VERY IMPORTANT.
AND AS FAR AS THE WEBINARS WE
COME TOGETHER TO TRY AND DECIDE
THESE THINGS AND HOW WE'RE GOING
TO DO IT AND WHO THE SPEAKERS
ARE GOING TO BE AND THERE'S
CONFIRMATIONS CONTRACT WITH THE
ADVOCATORS AND MAKE THE TRAIN
RUN MAKE IT HAPPEN.
AND IT'S A COLLABORATION WITHIN
DARE AND THE MDC COLLABORATORY.
IS THAT SEVEN MINUTES?
>> THANK YOU.
I'D LIKE TO ASK QUESTIONS OF THE
DARE TEAM.
>> QUESTION FOR LOUIS.
I REALLY LIKED SEEING THE
RESPONSE IN THE mRNA GROUP
COMPARED TO THE CONTROL GROUP
AND DO YOU SEE RESPONSES TO THE
CTL PRESSURE?
>> YOU MEAN THE SEQUENCE THE
VIRUS POST ART RELEASE?
>> YES.
>> WE HAVEN'T DONE THAT YET.
THAT'S SOMETHING ON OUR LIST.
>> I HAD A COUPLE QUESTIONS.
TERRIFIC PRESENTATIONS.
FIRST FOR LOUIS, WERE YOU ABLE
TO IDENTIFY A BIOMARKER THAT WAS
CORRELATED WITH THE DELAYED
REBOUND OF NINE DAYS WITH THE
mRNA VACCINE?
IN OTHER WORDS, DO YOU THINK THE
LOWER LEVELS OF VIRUS EXPRESSING
CELLS AND TISSUES WITH THE
VACCINATION STRATEGY IS WHAT
ACCOUNTED FOR THE DELAY IN
REBOUND OR DIRECTLY RELATED TO
HIV SPECIFIC OR HIV SPECIFIC CD
8 T CELLS?
>> THE ONLY WAY TO ANSWER THAT
IS REALLY TO SEE CD 8 T CELL
DEPLETION AND THE VACCINE WAS
OTHERWISE IDENTICAL.
WE'RE COMPARING IT TO THE
CONTROL VACCINE.
SO THE INMATE IMMUNE RESPONSES
SHOULD BE THE SAME.
I THINK THE DATA STRONGLY
SUPPORT THIS IS THE ADAPTIVE
RESPONSE ELICITED BY THE VACCINE
WHICH I MEAN THE VACCINE DOES
MAKE ANTIBODIES BUT THE
ANTIBODIES ARE TO GAG SO THERE'S
ALSO CD4 T CELLS THERE YOU CAN'T
RULE OUT THEY DON'T HAVE AN
EFFECT BUT THE MOST LIKELY
PREDICT IF WE DEPLETE CD T CELLS
WE'D LOSE THE DELAY AND CONTROL.
>> YOU ALLUDED TO CONTROL BEING
ASSOCIATED WITH LOW RESERVOIR
SIZE AND WONDER IF YOU CAN GIVE
US A QUANTITATIVE RANGE FOR WHAT
YOU CONSIDER LOW SIZE IN YOUR
COHORT.
>> YEAH, SO WHEN I SAY LOWER
RESERVOIR SIZE THE ONLY
MEASUREMENTS HAVE BEEN DNA WITH
IPDA INTACT MEASUREMENTS AND
CELL RNA AND THE RESERVOIRS I'M
NOT RECALLING -- LIKE I CAN'T
GIVE EXACTLY THE NUMBER BUT WHEN
WE COMPARED THE IN TACT DNA FROM
EARLY TREATED PEOPLE THEY FELL
IN THE RANGE OF PEOPLE WHO
TYPICALLY WHO RECEIVED EARLY ART
EARLY AFTER HIV ACQUISITION.
SO I CAN LOOK AT THAT WITH YOU
LATER TO GIVE YOU A PRECISE
NUMBER.
>> TEN OR A HUNDRED OR A
THOUSAND.
>> SO, MANY OF THE NUMBERS ARE
EVEN LOWER THAN SO AND AT TIMES
UNDETECTABLE AND OTHER TIMES
DETECT
DETECTABLE.
THE ONE INDIVIDUAL THAT DIDN'T
REBOUND AND SEVERAL MEASUREMENTS
UNDETECTABLE AND IN OTHER
STUDIES WITH DIFFICULT TO ELICIT
PRODUCTIVE INFECTION IN S.
WE DIDN'T DO THAT IN THE STUDY.
>> AND THE THIRD QUESTION IS FOR
SHARON.
REALLY AMAZING WORK ON THE SMAC
MIMETICS AND I THINK I REMEMBER
A SIMILAR EFFECT IN INHIBITORS
WHERE IT DID IMPACT ON HIV CD 8
RESPONSES.
AND I WONDERED TO WHAT EXTENT
YOU THINK THIS IS A LARGER
PHENOMENON OF LATENCY REVERSAL
LEADING TO CD 8 T CELL
EXHAUSTION PERHAPS AND LOSS IN
THE SETTING.
>> YEAH, YOU'RE RIGHT.
THEY WERE SHOWN TO AFFECT
FUNCTION ACTUALLY.
I THINK IT'S A CONSEQUENCE OF
THESE NOT BEING HIV SPECIFIC.
THEY HAVE OFF TARGET EFFECTS.
I THINK THE MECHANISMS OF WHY
INHIBITORS HIT AZD AND SMAC
MIMETICS ARE PROBABLY DIFFERENT.
I THINK IT'S A CONSEQUENCE OF
USING DRUGS SPECIFIC THAT
THEREFORE IMPACT CELL DEATH
PATHWAYS FOR AZD.
>> I THINK IT'S GREAT YOU WERE
ABLE TO COME TO A CONCLUSION IN
TERMS OF CONSIDERATION OF USE
THE MOLECULES.
WHAT WOULD YOUR RECOMMENDATION
BE?
CAN WE SAY GO, NO GO FOR THE
BASED ON THESE FINDINGS.
>> YEAH, I THINK OUR FINDINGS
AROUND THE ADVERSE EFFECT OF
SMAC MIMETICS ARE LIMITED TO
AZD5582 AND THERE'S BEEN EFFECTS
AS SHOWN BEFORE IN THE CANCER
LITERATURE.
AND WHETHER THIS EFFECT IS FOR
ALL BUY V -- BIVEIL -- BIVALENS
AND IT'S INTRIGUED ME WHY IT
LOOKS DIFFERENT IN VIVO AND EX
VIVO.
>> THANK YOU.
RICK DUNHAM.
THANK YOU.
GREAT SET OF STUDIES AND PRETTY
DISTINCT AND IT DOESN'T JIVE
FROM OTHER DATA SETS IN TERMS OF
ATM AFTER AZD TREATMENT SO
INTERESTED TO TALK THROUGH THE
DIFFERENCES AND SEE IF WE CAN
UNDERSTAND THIS AND FIGURE OUT A
WAY TO PUSH THIS FORWARD.
THE BIVALENCE IS VERY SHORT
MAYBE ONE HOUR FOR THESE
PERIODS.
IN VITRO WERE YOU SIMULATING
EXPOSURE TO THE DRUG?
>> I'M PRETTY SURE IT WAS
CONTINUOUS.
>> DO YOU SEE VERY DIFFERENT
OUTCOMES WITH SHORT EXPOSURES
AND TALK THROUGH THAT.
>> WE HAVEN'T LOOKED AT THAT BUT
IN IN VIVO I ASSUME YOU'RE
GIVING CONTINUOUS BECAUSE YOU'RE
REDOSING.
NO, SINGLE DOSE.
WE WERE DOING CONTINUOUS DOSING
IN THE EX VIVO STUDY.
>> MY NAME IS KARLY FROM THE
UNIVERSITY OF WASHINGTON.
MY QUESTION IS FOR SHARON AND
CURIOUS WHAT YOU THINK ABOUT THE
ALTERNATIVE HYPOTHESIS THAT
AZD5582 IS RESENSITIZING THE
CELLS LEADING TO A HIGHER SET
POINT AND NOT SURE IF THERE'S
WAYS TO ADD BACK IN THE HIV
SPECIFIC CD 8 CELLS TO FULLY
LINK THOSE CELLS TO THE
PHENOTYPE OR NOT.
>> SO I THINK YOU'RE REFERRING
TO THE LAST DATA THE PIECE OF
DATA WE SHOWED WHICH WAS A
HIGHER SET POINT IN THE MONKEYS
THAT RECEIVED THE CONTROL
VACCINE WITH AZD WITH CONTROL
WITHOUT AND THEY DO RECEIVE THE
CONTROL VACCINE.
I THOUGHT THAT TOO MAYBE YOU'RE
INCREASING THE POOL OF
TRANSCRIPTIONALLY ACTIVE CELLS
ONCE YOU STOP ATI AND
THEORETICALLY WE ALWAYS SAID
WHEN YOU REVERSE AGENTS ON ART
THAT DOESN'T EXPAND THE
RESERVOIR BECAUSE YOU EXPAND THE
VIRAL REPLICATION BUT YOU PLAY
BE INCREASING ALL THE CELLS
RECALL TO FIRE UP FAST.
THAT'S A POTENTIALLY REASONABLE
EXPECTATION.
HAVE WE ADDED BACK IN CTL?
NO.
BUT I THINK THERE'S MORE
MECHANISTIC STUDIES WE COULD DO
IN MONKEYS IF I CAN CONVINCE
LOUIS TO SPEND MORE ON THIS
MONKEY WORK WHICH IS ALWAYS HARD
TO CONVINCE BUT WE COULD
PROBABLY TEASE IT OUT MORE IN
MONKEYS.
>> I DON'T WANT TO CUT TOO MUCH
IN THE BREAK SO QUICK QUESTIONS.
>> SURE.
JUST FOR LOUIS CAN YOU COMMENT
ON CELL ASSOCIATED RNA WHICH IT
WAS ALTERED WITH THE DIFFERENT
VACCINE OR HAD ANY RELATIONSHIP
TO REBOUND RATES?
>> I THINK LOUIS WOULD KNOW THE
MONKEY STUDY I THINK WE SHOWED
SOME OF THAT.
>> IN MONKEYS ESPECIALLY MONKEYS
GIVEN THE DNA IT'S A GOOD
REFLECTION OF REPLICATION
COMPETENT VIRUS.
THE PEAK VIRAL LOADS WERE
IDENTICAL IN BOTH GROUPS AND SO
THEN WE DIDN'T SEE DIFFERENCES
IN CELLS ASSOCIATED DNA IN THE
ART VACCINE.
>> LOUIS ASSOCIATED WITH AND
WITHOUT AZD IN THE VACCINATED
GROUP.
DO YOU REMEMBER THAT?
>> I DON'T REMEMBER THERE BEING
DIFFERENCES.
THE AZD WAS ANOTHER STUDY.
>> WE HAVE THAT DATA AND I'LL
CHECK IT FOR YOU BECAUSE IT WAS
DONE IN ALL FOUR ARMS.
WE CAN'T SHOW YOU AZD VERSUS NOT
BUT WE HAVE THAT.
>> MAYBE THIS WILL CARRY INTO
THE BREAK BUT THANKS FOR
PRESENTING THIS WORK.
IT WAS INTERESTING AND AS I'M
SURE YOU KNOW WE HAVE QUITE
DIFFERENT RESULTS WHEN WE TREAT
IN VIVO NON HUMAN PRIMATES WITH
SMAC MIMETICS AND WHEN WE
PUBLISHED IN NATURE AND MEDICINE
LAST YEAR WE FOUND THE
COMBINATION WITH HIV SPECIFIC
ENVELOPE TARGETING ANTIBODIES
ACTUALLY LED TO AN INCREASE IN
SIV GAMMA PRODUCING T CELLS SO
MAYBE RELATED TO WHAT YOU'RE
COMBINING AZD IN TERMS OF THE
EFFECTIVE ANTIBODIES AND THE
EFFECT OF THE mRNA VACCINE AND
I'M WONDERING IF YOU HAVE
MECHANISTIC DATA IN TERMS OF THE
RESULTS IN TERMS OF THE EFFECT
ON THE CD 8 T CELLS AND WHAT THE
SMAC MIMETIC WAS CHANGING TO
HELP US TEASE OUT DIFFERENCES
BETWEEN MULTIPLE EXPERIMENTS.
>> I THINK IT'S A GOOD POINT.
DEPENDING ON WHAT CYOUR CONTROL
AND THE T CELL VACCINE AND SAW
ABROGATION OF THE EFFECT AND MAY
NOT TRANSLATE TO OTHER
INTERVENTIONS AS YOU'VE
PUBLISHED WITH ANTIBODIES.
WE HAVE GOT SOME SINGLE CELL RNA
DATA AND GOING THROUGH THAT TO
UNDERSTAND THE EFFECT.
IT MAY BE MY CURRENT HYPOTHESIS
IS THE EXPRESSION OF PROTEINS IN
THE SPECIFIC T CELLS MAKING THEM
VULNERABLE TO CELL TOGETHER BUT
THAT'S PRELIMINARY.
>> THE WE SEE A BENEFICIAL
EFFECT THE VIRAL LOADS ARE LOWER
DURING TREATMENT.
YOU DO THE SAME WITH VACCINE AND
DO THE OPPOSITE AND HARM THE
OUTCOME OPPOSED TO HELPING.
AND WE BOOST RESPONSE THAT IS
SHOWING VULNERABILITY THAT MAY
NOT BE THE SAME IN ALL YOU'VE
DONE.
>> GREAT, THANK YOU.
LET'S GIVE A ROUND OF APPLAUSE.
AND THANK YOU SO MUCH.
RIGHT NOW WE HAVE A BREAK.
LOSE ENJOY THE COFFEE SPONSORED
BY THE COLLABORATORY P.I.s AND
SNACKS PERFECT -- ARE PROVIDED
BY THE PROGRAM OFFICERS.
WE'LL RECONVENE PROMPTLY AT 11:00.
SINCE WE ARE ABOUT HALFWAY
THROUGH WE'VE HAD FIVE NDCs AND
HAVE FIVE LEFT.
I JUST WANT TO REMIND EVERYONE
TO BE COGNIZANT OF THE NIAID
LANGUAGE GUIDE.
INSTEAD OF SUBJECT WE CAN SAY
PARTICIPANT OR INFECTED
INDIVIDUAL PEOPLE LIVING WITH
HIV.
WITH THAT I'D LIKE TO INTRODUCE
THE BEAT HIV COLLABORATORY LEAD
BY THE LUIS AND JIM AT U-PEN AND
JOHN AT JOHNS HOPKINS.
WITH THAT I'LL LET LUIS TAKE IT
AWAY.
>> THANK YOU.
IT'S EXCITING TO SHARE WITH YOU
THE PROGRESS THE GROUP HAS BEEN
MAKING AND IT WAS HARD TO CHOOSE
THE TALKS WE'RE GOING TO PRESENT
BUT THEY'RE ALL REALLY EXCITING
AND REALLY IMPACTFUL ACROSS THE
WORK OF ALL THE COLLABORATORIES.
THE COLLABORATORY AS A WHOLE
STILL MAINTAINS AN ACTIVE GROUP
OF PEOPLE.
THANK YOU FOR PARTICIPATING IN
THE WORKING GROUPS.
AND WE'RE FOCUSSED ON THE FOUR
CENTRAL SEN -- TENETS IN
STRATEGY FOR CONTROL OR
ERADICATION.
ONE THING OUR COLLABORATORY IS
DISTINCT FOR IS THE EMPHASIS ON
THE SOCIAL SCIENCE AGENDA.
WE'VE DEVELOPED THE CAB WITH
SOCIAL SCIENCE AND ONE PROJECT
HAS JUST BEEN ACCEPTED FOR
PUBLICATION AND WE'LL HEAR ABOUT
IT TODAY.
AND PROGRAMS FEATURED TODAY AND
TODAY AND WE KEEP A CLOSE TRACK
RECORD OF THE COLLABORATORY AND
TRYING TO MAKE CERTAIN YOU HOW
INTEGRATIVE THE COLLABORATIVE IS
AND THE GRANTS THAT EMERGE.
I THINK THE HIGHLIGHTS ARE HERE
AND THE LATENT RESERVOIR IS
BUILDING AN INVENTORY OF
INDIVIDUALS WITH APHERESIS AND
TIME POINTS AND EXPANDING THE
TISSUE ACCESS TO THESE
INDIVIDUALS.
THE BIOLOGICAL AGING ASPECT AND
BIOMARKER WORK CONTINUES TO MOVE
FORWARD AND THE FOCUS ON
STRATEGIES.
T
THE EVENT OF STRATEGIES WE DON'T
HAVE TIME TO PRESENT ALL BUT
THEY'RE ALL EXCITING AND
DEVELOPING NOVEL STRATEGIES IN
EACH ONE OF THESE TO HAVE A TALK
ON EACH ONE OF THEM AS TIME
WOULD ALLOW.
BUT MY TANK IS TO BE QUICK
BECAUSE WE HAVE FOUR STORIES I
WANT TO ALLOW TIME FOR BUT AS
THE OTHER COLLABORATORIES
CONTINUE TO EVOLVE AND THESE ARE
FUNDING AND CONTRACTS THAT HAVE
COME UP HIGHLIGHTING THE
EVOLUTION OF STRATEGIES AND NEW
INVESTIGATORS COMING ON BOARD
WITH THE COLLABORATORY.
TWO OF WHICH WE'LL HERE FROM
TODAY AS NEW INVESTIGATORS THAT
ARE NOW SUPPORTED BY THE
COLLABORATORY.
WE'RE ALSO EXCITED TO ENGAGE IN
AN INITIATIVE TO OPEN A NEW HIV
CURE AND VIRAL CENTER AND
EVALUATING PROGRAMS SO IF YOU
KNOW ANYONE INTERESTED IN
DEVELOPING THIS AREA AND
RELOCATED SEE ME AFTERWARDS.
FOR TODAY WE'LL TALK ABOUT TWO
ESTABLISHED PROGRAMS AND TWO NEW
PROGRAMS.
AND THESE TWO TALKS WE'LL HEAR
ABOUT FROM BOB AND MIKE ARE
REALLY CENTERED ON AREAS UNDER
STUDIED OR OVERLOOKED.
AS THE DISCUSSION MOVES FORWARD
YOU'LL SEE WHAT I MEAN REGARDING
THE POWERFUL ANTIBODIES THE
TEAMS AT PENN ARE CHASING AND
OUR CONTINUED DESIRE TO
UNDERSTAND WHERE IS THE
PERSISTENT VIRUS AND HOW BEST TO
MEASURE IT AND THE
PROPORTIONALITY OF THE RESERVOIR
BETWEEN THE SUBSETS OF THE CD4
CELL.
WITH RESPECT TO NEW PROGRAMS
COMING THERE'S AN EXCITING
DEVELOPMENT FROM CAR T CELL AS
NEW THERAPY APPROACH THAT SHOW
THE POTENCY OF BROADLY
NEUTRALIZING ANTIBODIES AS THE
STRATEGY FOR DETECTION AND
DR. CLAIBORNE WILL TELL US ABOUT
THAT.
ANOTHER UNDER STUDIED SOCIAL
SCIENCE SIDE IS WE TALK ABOUT
OUTREACH TO THE COMMUNITIES AND
WE HAVE THE PROVIDERS WHICH ARE
GATE KEEPERS TO THE PEOPLE
LIVING WITH HIV WHEN THEY LEARN
AND TRY TO SIPHON AND UNDERSTAND
WHAT'S DEVELOPING WHETHER IT'S A
DEVELOPMENT AND HOW BEST TO
ENGAGE THE COMMUNITY FOR THE
BENEFIT OF MOVING THE SCIENCE
FORWARD.
SO WITH THAT, I WE'LL DO A
VIRTUAL TRANSFER TO BOB AND
AFTER THAT MIKE AND THE LAST ONE
WILL BE VIRTUAL IF WE CAN
TRANSFER THE PRESENTATION TO HIS
LAPTOP.
WE PRACTICED THIS DURING THE
BREAK SO WE SHOULD BE GOOD.
>> CAN YOU SEE MY SLIDES?
>> YES.
I'LL TALK ABOUT AUTOLOGOUS
ANTIBODY.
PEOPLE WITH HIV MAKE A RESPONSE
AND THEY NEUTRALIZE THE VIRUS
AND WE ARE EXPLORING THE ROLE OF
THE ANTIBODIES IN HIV STRATEGIES
AN IN SOME SITUATIONS THEY MAY
BE ABLE TO MEDIATE A FUNCTIONAL
CURE.
NOW WE BECAME INTERESTED IN THE
ANTIBODIES THROUGH THE
CONSIDERATION OF THE CLONAL
NATURE OF THE HIV RESERVOIR.
IT'S CLEAR REPEATED EXPOSURE TO
THE AIM ANTIGEN CAN GENERATE
LARGE CLONES OF HIV INFECTED
CELLS TRANSFORMING THE RESERVOIR
FROM A COLLECTION OF QUIESCENT
CELLS DOMINATED BY CLONES AND
REBOUND AND INTERRUPTION WILL
COME FROM THE LARGE CLONES.
INTERESTINGLY, TWO RECENT
STUDIES HAVE SHOWN EVEN WHEN THE
RESERVOIR IS DOMINATED BY A
SINGLE CLONE TREATMENT AND
INTERRUPTION COMES FROM
PHYLOGENETICALLY DISTINCT
VARIANTS.
HERE'S A STUDY BY MICHEL.
IN THINKING ABOUT THIS IT'S
USEFUL TO CONSIDER THE RESERVOIR
ON THE LOG SCALE SHOWING THE
DIFFICULTY IN REDUCTION GIVEN
RAPID EXPONENTIAL VARIANTS CAN
COME FROM A SINGLE CELL.
WE NEED TO EXPLAIN WHY DOESN'T
REBOUND COME FROM THE DOMINANT
CLONES?
HYPOTHESES WERE PUT FORWARD AND
WE FAVORED A SIMPLER ONE THE
LARGE CLONES CANNOT CAUSE
REBOUND BECAUSE THE VIRUS IS
ALREADY NEUTRALIZING BY THE
AUTOLOGOUS ANTIBODY RESPONSE AND
IT'S TYPE SPECIFIC AND RESULTS
SHOW IT'S EFFECTIVE AND MAY HOLD
THE KEY TO A CURE.
NOW, TO TEST THE HYPOTHESIS
AUTOLOGOUS NEUTRALIZING
ANTIBODIES CAN PREVENT REBOUND
FROM THE LARGE CLONES WE CARRIED
OUT LARGE GROWTH ASSAYS IN THE
PRESENCE AND ABSENCE OF LOW
CONCENTRATIONS OF AUTOLOGOUS
CONTEMPORANEOUS IGG ANTIBODIES
AND SHOWED ONLY A SUBSET OF
RESERVOIR VIRUSES COULD GO OUT
IN THE PRESENCE OF AUTOLOGOUS
IGG.
THE VIRUSES ACCUMULATED
MUTATIONS IN THE FIVE VARIABLE
REGIONS OF THE ENVELOPE GENE.
IF WE LOOK AT THE ENVELOPE
TRIMER THEY'RE LOCATED IN THE
PART OF THE TRIMER AND LOOKING
FROM THIS PERSPECTIVE YOU CAN
SEE THE VARIABLE REGIONS
ENCOMPASS THE TRIMER AND THE
REGIONS ARE BOUNDED BY CYSTEINE
RESIDUES BOUND TO EACH OTHER AND
THE RESIDUE BETWEEN THE
CYSTEINES CAN VARY IN NUMBER,
SEQUENCE AND GLYCOSYLATION AND
IT'S LIKELY THE CAPACITY FOR
VARIATION EVOLVED TO ALLOW
ESCAPE FROM THE AUTOLOGOUS
NEUTRALIZING ANTIBODY RESPONSE
WHICH NORMALLY TARGETS THIS
REGION.
IT'S THE AND THE ASSAYS SHOWED
THE VIRUSES GROWING OUT IN THE
PRESENCE OF AUTOLOGOUS IGG WERE
HIGHLY RESISTANT AS EXPECTED BUT
OTHER VIRUSES FROM THE SAME
DONOR IN THE LARGE CLONES WERE
HIGHLY SENSITIVE AND LYNN WENT
ON TO SHOW THE VIRUSES THAT ARE
RESISTANT ARE CONNECTED TO VIRUS
IN REBOUND IN VIVO AND IT'S
STILL EFFECTIVE IN PREVENTING
REBOUND FROM A SUBSET OF
RESERVOIR VIRUSES.
THE FRACTION OF VIRUS NEUTRALIZE
VARY TREMENDOUSLY FROM PERSON TO
PERSON.
THE FRACTION OF VIRUS NEUTRALIZE
CORRELATE WITH TIME TO VIRAL
REBOUND AND IN A STUDY AFTER
ACCOUNTING FOR BNAB WASH OUT.
THEY'RE STABLE AND SHOWING
SENSIT
SENSITIVE ISOLATES FROM DONORS
ON ART OVER 20 YEARS.
SO REBOUND IS A SELECTION
PROCESS.
ANTIGENS ARE CONSTANTLY
ACTIVATING RESERVOIR CELLS AND
GIVES RISE TO THE RESIDUAL
VIREMIA.
MANY HAVE A RESPONSE AND REBOUND
REQUIRES ACTIVATION OF A CELL
CARRYING A NEUTRALIZATION
RESISTANT VIRUS.
ONLY A SUBSET OF RESERVOIR
VIRUSES CAN CAUSE REBOUND.
NOW, THE EVOLUTION OF ESCAPE
VARIANTS IS A PROCESS THAT
PREDATES AND ORTHOGONAL TO
CLONAL EXPANSION.
IN SOME DONORS, THE LARGE CLONES
ARE ACTUALLY NEUTRALIZATION
RESISTANT.
NEVERTHELESS THE FINDING AN
ANTIBODY RESPONSE GENERATED
YEARS AGO CAN STILL PREVENT
REBOUND FROM A SUBSET OF
RESERVOIR VIRUSES IS FROM RECENT
YEARS.
IT CAN BE ANALYZED IN THE
MACAQUE MODEL AND WE HAVE AN
ADVANTAGE TO KNOW WHAT THE
STARTING CONSEQUENCE SEQUENCE
AND HERE'S A SINGLE ANIMAL OVER
36 WEEKS PRIOR TO THE INITIATION
OF ART.
THE VIRAL LOAD IS EXTREMELY HIGH
AT THIS TIME.
AT THE TIME OF INITIATION THE
VIRUS IS REPLICATING IS IN BLACK
AND ACCUMULATED MUTATIONS IN THE
ENVELOPE GENE CONSISTENT WITH
ESCAPE FROM AUTOLOGOUS
NEUTRALIZING ANTIBODIES.
ANCESTRAL VARIANTS WITH FEWER
MUTATIONS ARE PRESERVED IN THE
RESERVOIR AND ABSENT FROM THE
PLASMA AT THE TIME OF VARIATION
DESPITE FITNESS SHOWN BY THE
VIREMIA EARLY ON AND THIS LIKELY
REFLECTS THE NEUTRALIZATION BY
AUTOLOGOUS NEUTRALIZING
ANTIBODIES.
THIS SUGGESTS A CURES STRATEGY.
INSTEAD OF REDUCING THE
RESERVOIR BY MANY LOGS, WHICH WE
DON'T KNOW HOW TO DO, WE CAN
SUPPLY OR GENERATE ANTIBODIES TO
THE ANAB RESISTANT SUBSET OF
VIRUSES.
CAN THEY PRODUCE A FUNCTIONAL
CURE?
AND I'D LIKE TO CONCLUDE WITH A
CASE THAT SUGGESTS THE ANSWER
MAY BE YES.
THIS IS A PARTICIPANT IN A STUDY
OF A 45-YEAR-OLD MALE STARTED
ART TWO MONTHS AFTER INFECTION.
AT THAT TIME THE VIRAL LOAD WAS
VERY HIGH AND NO PROTECTIVE HLA
ALLELES.
HE RECEIVED AN LRA AND BNAB AT
THE TIME OF INITIATION AND AFTER
ONE YEAR IT WAS STOPPED AND
REBOUND DID NOT OCCUR AND IT'S
NOW BEEN OVER SIX YEARS WITHOUT
A VIRAL REBOUND.
THIS IS DESPITE THE PRESENCE OF
REPLICA
REPLICATION COMPETENT RESERVOIR.
AND WE ASKED WHETHER THE CONTROL
COULD BE DUE TO AUTOLOGOUS
NEUTRALIZING ANTIBODIES.
THIS IS AN HIV NEGATIVE DONOR.
AT THE TIME OF INITIATION
THERE'S NO NEUTRALIZING ACTIVITY
IN THIS PARTICIPANT'S PLASMA BUT
AFTER THREE MONTHS STRONG
NEUTRALIZATION IS DETECTED AND
AFTER SIX MONTHS AND ALL
SUBSEQUENT TIME POINTS THERE'S
VERY STRONG NEUTRALIZING
ACTIVITY.
THE IC50 IS ONE MICROGRAM PER
MIL AND ONLY A MINUTE FRACTION
OF WHICH IS COMPRISED OF HIV
SPECIFIC ANTIBODIES.
AND I'LL ALSO POINT OUT THE
NORMAL PLASMA IGG CONCENTRATION
IS 10 MISSION -- MIGS PER MIL
AND WE CAN COMPARE THE
NEUTRALIZING ACTIVITY IN THIS
PARTICIPANT'S PLASMA TO ART
REGIMENTS AND THEY INDIVIDUALLY
PRODUCE LESS THAN 3 LOGS OF
INHIBITION WHEREAS EFFECTIVE
REGIMENTS PRODUCE ABOVE 5 LOGS
OF INHIBITION.
AND LOOKING AT THE AUTOLOGOUS
NEUTRALIZING ANTIBODIES IN THIS
PARTICIPANTS PLASMA ONCE THEY
ARE ARISE THEY ACHIEVE A LEVEL
OF INHIBITION COMPARABLE TO ART.
BY THE TIME THE ANTIBODIES AROSE
THE PARTICIPANT WAS ALREADY ON
ART SO THERE WAS NO CHANCE FOR
ESCAPE MUTATIONS.
AND INTERESTINGLY IF YOU LOOK AT
THE ANTIBODIES FOR PEOPLE WITH
HIV THERE'S NO ACROSS
NEUTRALIZING.
THE ANTIBODIES ARE LIKELY
DIRECTED AT THE VARIABLE
REGIONS.
SO WORKING WITH DUKE WE HAVE NOW
OBTAINED A CRYO EM STRUCTURE OF
THE ENVELOPE FROM THIS
PARTICIPANT AND WE'RE PULLING
OUT THE ANTIBODIES IN HOPES WE
CAN LEARN FROM THIS EXTREMELY
POTENT BUT HIGHLY SPECIFIC
RESPONSE.
SOME INSIGHTS TO HELP DEVELOP A
GENERAL CURE STRATEGY.
MY WIFE AND I HAVE LAD THE
PLEASURE OF WORKING WITH A
REMARKABLE GROUP OF GRADUATE
STUDENTS AND MENTIONED THEM IN
THE WORK AND WOULD LIKE TO THANK
MANY COLLABORATORS FROM THE
COLLABORATORIES AND A GROUP FROM
DUKE UNIVERSITY.
>> THANK YOU, EVERYBODY.
I'M HAPPY TO BE HERE IN MULTIPLE
WAYS TO PRESENT THIS WORK TO YOU
FROM OUR LAB THROUGH THE B
KWLABTRY.
WHAT I' -- COLLABORATORY.
AND WHAT WE'LL START WITH THE
PLAIN LANGUAGE IS WHERE IS HIV?
WHAT I MEAN IS WHAT TYPE OF CELL
IS HARBORING HIV IN PEOPLE
LIVING WITH HIV AND TISSUES
WHERE WE KNOW THE PREDOMINANCE
OF THE RESERVOIR RESIDE AND WHAT
CELLS ARE IN LYMPH NODE AND IS
THE VIRUS LATENT IN TISSUES
DURING ART.
WE FOUND THE HIV RESERVOIR IN
LYMPH NODES HAVE DIFFERENT
PROPERTIES AND MAY NOT ENTIRELY
BE LATENT AND WHY IS THIS
IMPORTANT?
AS WE THINK OF DIFFERENT TYPES
OF CELLS MAY PRESENT UNIQUE
CHALLENGES FOR CURE ESPECIALLY
WHEN THE CELLS WITH HIV MAY NOT
DIFFER VERY MUCH HENCE THE
WHERE'S WALDO PICTURE.
THE STUDY GOAL WAS TO DEFINE THE
HIV RESERVOIR AND TREAT PEOPLE
WITH HIV.
WE WANTED TO LOOK AT THE
TRANSCRIPTOMIC AND EPITOPE IN
LYMPH NODESES OF PEOPLE WITH ART
AND USED A TECHNIQUE FIRST USED
IN THE CONTEXT OF HIV CALLED
DOGMA SEQ PUBLISHED I GUESS LATE
LAST YEAR.
WE WANTED TO DETERMINE WHERE THE
PREDOMINANT CELL SUBSETS
HARBORING IN THE ART TREATED
LYMPH NODES AND THE MOST THING
YOU'LL THINK ABOUT IS HELPER
CELLS.
AND WE WANT THE TO FIGURE OUT IF
THAT WAS REALLY THE CASE.
WE WANTED TO DETERMINE IF THE
CD4 T CELLS HARBORING HIV WERE
QUIESCENT OR TRANSCRIPTIONALLY
ACTIVE AND IF THERE WERE
DIFFERENT CHARACTERISTICS
THEREOF AND IN THIS PRESENTATION
WE WON'T BE ABLE TO GET TO THE
LAST POINT DUE TO TIME.
WE USED A TECHNIQUE CALLED DOGMA
SEQ I WON'T GO THROUGH THE
ENTIRE PROCEDURE OUT OF TIME.
THE MAIN POINT TO TAKE HOME ARE
ESSENTIALLY WHAT WERE THE CELLS
WE STARTED WITH AND SO PAY
ATTENTION TO THE TOP PART OF
THIS FIGURE WHERE WE INSTEAD OF
USING BEAD MEDIATED SELECTION AS
DONE TO PURIFY CELLS WE USED
FLOW SORTING TO LIVE SORT CELLS
AND PARTICULARLY TO INCLUDE CD
5RA IN THE SELECTION.
WE HAD SAMPLES I'LL DESCRIBE IN
THE NEXT SLIDE FROM INDIVIDUALS
OFF VEREMIC CHRONIC INFECTION
AND THOSE ON THERAPY.
AND WE DIFFERENTIALLY CHOSE
CELLS BASED ON THE FREQUENTED
CELLS FROM HIV.
THE CHRONIC SAMPLES WE USED A
LITTLE OVER 25,000 CELLS FOR
DONOR AND FOR THE ART A LITTLE
OVER 50,000 CELLS PER DONOR THE
TOTAL CELLS WE ANALYZED THAT
PASSED WERE GREATER THAN 400,000
SINGLE CELLS AND WE HAVE
ADDITIONAL DATA NOT ADDED TO
THIS WE'VE BEEN ANALYZING AS WE
HAVE BEEN SITTING IN HERE THAT
BRINGS US TO ABOUT 650,000
SINGLE CELLS.
BUT THE CONCLUSIONS HAVE NOT
CHANGED FORTUNATELY.
AND THESE ARE INDIVIDUALS LIVING
WITH HIV EITHER TREATED OR
UNTREATED WHO DONATED SAMPLES
TYPICALLY CERVICAL LYMPH NODES
FOR DIAGNOSTIC PURPOSES.
YOU CAN SEE WE HAVE 10
INDIVIDUALS ON ART, 10
INDIVIDUALS WHO HAD NOT YET
BEGUN ART AT THE TIME OF THE
LYMPH NODES BIOPSY AND SUFFICE
TO SAY WE WERE ABLE TO CLEARLY
DEFINE AROUND 23 OR SO DIFFERENT
CLUSTERS OF CELLS FROM THE LYMPH
NODES OF THE INDIVIDUALS AND
COMPRISED SUBSETS AS DETAILED
HERE.
SO IMPORTANTLY FOR EXAMPLE TWO
DIFFERENT SUBSETS OF HELPER
CELLS AND I WON'T GO THROUGH THE
REST.
MANY DIFFERENT SUBSETS OF CELLS
AND WE'LL HAVE MORE IN A FEW
MOMENTS.
THE KEY ISSUE IS COULD WE FIND
HIV CELLS HARBORING HIV AND WE
CAN DETECT THOSE BOTH BY
DETECTING VIRAL DNA AND RNA
AFTER ALIGNING THE SEQUENCING
DATA.
WE'RE ABLE TO FIND DNA AND RNA
POSITIVE SAMPLES IF YOU LOOK AT
THE LEFT SIDE OF THE SLIDES IN
BOTH THE ART TREATED AND CHRONIC
INDIVIDUALS WE OBTAINED 400
SINGLE CELLS IN THE ART TREATED
INDIVIDUALS AND ABOUT 1200 OR
MORE -- SORRY, 1300 OR MORE FROM
THE CHRONIC INDIVIDUALS AND
WE'RE ABLE TO DETECT DNA AND RNA
POSITIVE CELLS AND YOU CAN SEE
FROM THE MAPS THE RED CELLS OVER
THE U MAPS ARE THE CELLS THAT
HAVE DETECTABLE PROVIRAL OR RNA
READ AND WE MAY NOT BE ABLE TO
LOOK AT THE MODALITY AND THE
DEFICIENCY IS ABOUT 50%.
WHAT ARE THE TAKE-HOME MESSAGES
FROM THE DATA SET?
I WON'T GET INTO DIFFERENTLY
EXPRESSED GENE ANALYSIS AND LOOK
AT SUBSETS HARBORING HIV.
WE'LL TALK ABOUT THESE IN THREE
GROUPS.
FIRST, FOCUSSING ON THE VIREMIC
INDIVIDUALS.
THE FIRST AND WHAT THE PLOT IS
SHOWING HERE IS IF WE TAKE ALL
OF THE IN THIS CASE PROVIRAL DNA
POSITIVE CELLS SET THAT TO 100%
AND ASK WHERE ARE THEY FOUND
WITHIN THE SUBSETS WE DEFINE.
THAT'S THE PROPORTIONALITY AND
SEE FIRST IT'S VERY
HETEROGENEOUS.
MANY CD4 SUBSETS IN THE LYMPH
NODE PLAY HAVE A PROVIRAL AID.
SO ABOUT WHAT ARE THE KEY THINGS
WE THINK ABOUT?
FIRST WE THINK OF CELLS AND
IDENTIFIED TWO TFH WE CAN GET TO
IN THE Q&A TO UNDERSTAND THE
DIFFERENCES AND IN COMBINATION
THEY ACCOUNTED FOR NEARLY 17% OF
THE TOTAL PROVIRAL DNA CELLS.
THEY WEREN'T THE LARGEST
POPULATION.
LARGEST WAS JUST A BORING
CENTRAL MEMORY CD4 POPULATION
ABOUT 35%.
THIS POPULATION HAS INTERESTING
CHARACTERISTICS.
I CALLED IT BORING BUT IT'S NOT
THAT BORING.
ONE OF THE TOP DIFFERENTLY
EXPRESSED GENES NEGATIVELY RECTS
TGF BETA YOU HEARD ABOUT IN THE
TALKS.
THERE'S ANOTHER MARKER CALLED OX
40 INDUCED UPON ACTIVATION.
THIS LOOKS LIKE IT MAY HAVE
ACTIVATION MARKERS.
IT'S ALSO NOT A RESIDENT
POPULATION SO IT'S A POPULATION
THAT WILL LEAVE THE LYMPH NODES
OUT TO THE BLOOD AND COME BACK.
THE LITERATURE HAS BEEN MIXED
WHETHER THE T CELLS ARE A MAJOR
CONTRIBUTOR TO THE RESERVOIR
UNDER ART.
IN OUR CASE FOR OUR SAMPLES WE
FIND IT IS NOT.
IN THESE SAMPLES ONLY 8% OF THE
TOTAL RESERVOIR WAS FOUND TO BE
IN CELLS DEFINED AS HELPER
CELLS.
WE FOUND TWO OTHER MEMORY
POPULATIONS THAT HIGHER
PROPORTIONS OF PROVIRAL DNA.
ONE WAS A NON-ACTIVATED CIRCUIT
POPULATION A CD4 CELL AGAIN THAT
WILL GO TO THE BLOOD.
AND THEN WE FOUND ANOTHER
POPULATION WHICH HAS NOT BEEN
DEFINED THAT WE KNOW OF AND IT'S
UNFORTUNATELY THE AN PLACE IS
CUT OFF THE DESCRIPTION OF IT
AND THE A41 POSITIVE POPULATION
I'LL TALK ABOUT IN A MOMENT
BECAUSE IT TURNS OUT TO BE AN
INTERESTING SUBSET.
BEFORE I TALK ABOUT THAT THOUGH
LET'S GO TO THE THIRD SMALL
STORY WHICH IS WHAT ABOUT THE
TRANSCRIPTIONALLY ACTIVE CELL?
I'VE TALKED ABOUT THE DNA ACTIVE
CELLS AND NOW THE RNA POSITIVE
CELLS.
ONLY IN THE ART TREATED
INDIVIDUALS THE CHRONIC
INDIVIDUALS COULD BE A FOURTH WE
WON'T TALK ABOUT BUT THE
QUESTION WAS WHAT ARE THE HELPER
CELLS DOING IN THIS CASE?
ONLY 5% OF THE RNA POSITIVE
CELLS ARE COMING OUT OF THE T
HELPER CELLS.
AGAIN IT'S THE POSITIVE
POPULATION THAT'S THE DOMINANT
POPULATION OF THE NR4A1 AND 2
POPULATION.
THEY'RE APPROXIMATELY 10% OF THE
TOTAL CD4s IN THE TREATED LYMPH
NODES IRRESPECTIVE OF TREATED OR
UNTREATED.
WHAT ARE THE CELLS?
THEY APPEAR CENTRAL MEMORY CD 28
AND CD HIGH AND THEY DO NOT
DISPLAY MARKERS OF RESIDENCY.
THEY'RE CD 69 LOW AND GO FROM A
LYMPH NODES TO THE BLOOD.
THEY APPEAR TO BE RESTING.
SO THEY'RE HLA NEGATIVE AND CD
38 NEGATIVE AND CD 3 LOW WHICH
SUGGEST THEY COULD BE ACTIVATED.
AND SUPPORTED BY THE TOP
EXPRESSED GENES.
THEY'RE TRANSCRIPTION FACTOR
THAT REQUIRES T CELL RECEPTOR
TRIGGERING WITHIN 3 TO 18 HOURS.
FROM MOUSE DATA AND COMMONLY
USED AS A REPORTER SYSTEM FOR
TCR ACTIVATION.
IT'S NOT INDUCED BY STANDARD
ACTIVATION.
IT'S A SPECIFIC EFFECT HAPPENING
BY CELLS GETTING TRIGGERED AND
THE RNA POSITIVE CELLS AND
THERE'S MULTIPLE FUNCTIONS
THAT'S A TRANSCRIPTION FACTOR
FACILITATE APOPTOSIS AND DRIVE
TOLERANCE.
AND THESE CELLS HAVE HIGH CD 69
RNA BUT HAVE LOW 69 PROTEIN OF
THE RNA TOE TO THE PROTEIN AND
APPEARS TO BE PHENOTYPICALLY
RESTING AND CELLS IN VIREMIA AND
ART.
I THINK I DON'T KNOW WHERE I AM
IN TERMS OF TIME.
I COVERED A LOT OF THIS.
THE RESERVOIR IS HETEROGENEOUS
IN ART.
WE SEE OTHER POPULATIONS THAT
PLAY HAVE LARGER CONTRIBUTIONS.
WE ARE SOMEWHAT LIMITED AND CAN
ONLY STAND FOR ONE OR TWO LYMPH
NODES AND THERE'S OVER 600 IN
THE BODY AND COULD BE
DIFFERENCES IN THE RESERVOIR
ACROSS THOSE TISSUES.
AND ONE REALLY IMPORTANT POINT
IS AND THIS HAS BEEN FOR EXAMPLE
MELANIE'S PRESENTATION SAYING
THE LATENT RESERVOIR IS NOT
TOTALLY LATENT THEY'RE
TRANSCRIPTIONALLY ACTIVE AND SEE
RNA POSITIVE CELLS THE VIRUSES
CONTINUALLY TRYING TO REACTIVATE
ALL THE TIME EVEN UNDER ART.
AND I'LL STOP THERE.
THE VAST MAJORITY OF ALL THIS
WORK WAS DONE BY VINCENT WU A
POSTDOC IN MY LAB AND I LIKE
WHEN PEOPLE SAY IT TAKES A
VILLAGE TO DO THIS.
IT TAKES OR CITY OR GLOBAL
AMOUNT OF FUNDING TO DO THESE
ASSAYS BECAUSE THEY'RE
INCREDIBLY EXPENSIVE SO THANK
YOU TO ALL THE FUNDING SOURCES.
>> ALL RIGHT.
SO, I'M EXCITED TO SHARE SOME
WORK THAT HAS BEEN A PROJECT
WE'VE BEEN DEVELOPING HEAVILY IN
THE PAST YEAR HALF LOOKING HOW
TO FURTHER DEVELOP T CELLS FOR
HIV.
FOR PLAIN LANGUAGE SUMMARY OUR
KEY QUESTION IS WHAT PARTS OF
HIV SPECIFICALLY THE ENVELOPE
ARE THE BEST TO TARGET AND WHY?
ENGINEERED T CELLS HAVE SHOWN TO
BE A VIABLE PART AND WE HAVE A
CAR T CELL AND ENVELOPE THAT CAN
BE TARGETED BY ANTIBODIES OR V
NABS AND DOES IT MATTER WHICH
ONE YOU TARGET AND WHY?
AND IT HAPPENS TO BE MORE DISTAL
ONES TOWARDS THE TOP OF THAT
TRIMER.
WE ALSO IMPLICATED CAR BINDING
THE STRENGTH WITH WHICH I BINDS
THE ENVELOPE TRIMER AS ONE
ASPECT DICTATING POTENCY AND
DEFINED COMBINATION OF CARS WE
THINK WE NEED IF WE'RE GOING TO
RESTRICT THE INCREDIBLE ABILITY
TO EVOLVE FROM ALL OF OUR
THERAPIES.
SO, AS I WAS SAYING WE WANT TO
KNOW WHAT PARTS OF THE HIV
TRIMER ARE THE BEST TO TARGET
AND WHY.
AND A LOT OF THE WORK WE'VE DONE
IN THE PAST HAS BEEN USING THE
PART YOU SEE ON THE LEFT.
IT'S A POTENT CAR T CELL AND HAS
GOOD IN VIVO SAFETY PROFILES AND
CAN'T ANSWER THE QUESTION
BECAUSE IT ONLY BINDS TO THE
BINDING SITE.
WE TOOK THE INCREDIBLE
LITERATURE AND FINDINGS FROM THE
BROADLY NEUTRALIZING ANTIBODY
FIELD AND MADE A LARGE PANEL OF
CAR T CELLS AND THAT ARE GET ALL
THE SIX MOST WIDELY DESCRIBED
AUTOLOGOUS NEUTRALIZING EPITOPES
AND THIS FORMS OUR TOOLBOX BY
WHICH WE CAN ASK QUESTIONS.
SO ONE OF THE FIRST AND SIMPLEST
THINGS IS HOW WELL DO THE
VARIOUS T CELLS KILL.
WE USED A KILLING ASSAY AND USED
TARGET RATIOS EXPRESSING HIV
ENVELOPE AND MONITOR REAL TIME
KILLING IN THE SYSTEM.
HERE YOU SEE THERE'S QUITE A
SPREAD AND THE ABILITY OF
VARIOUS CAR T CELLS TO KILL THE
TAR GETS AND IF WE DO THIS MANY
TIMES AND LOOK AT ACTUALLY FOR
VERY SPECIFIC EPITOPE CLASSES
HOW WELL ON AVERAGE THEY KILL WE
CAN SEE THERE IS HIERARCHY HERE.
IT'S NOT JUST A FLAT LINE.
CERTAIN ONES KILL BETTER OR NOT.
AND ONE THING WE NOTICE IS IF
YOU TALK THE RELATIVE DISTANCE
FROM THE PLASMA MEMBRANE AND
COMPARE THAT TO THE KILLING
SCORE IT DOES APPEAR THOSE MOST
DISTAL OR FURTHER FROM THE
ENVELOPE ARE ASSOCIATED WITH
ENHANCED KILLING POTENCY.
WE THEN WANTED TO CORROBORATE
THE FINDINGS BY PERFORMING A
MORE STRINGENT STRATEGY WITH T
CELLS AND REPLICATION COMPETENT
HIV AND MIX TARGET RATIOS WITH
CAR T CELLS AND IT'S MEASURING
THE CULTURE BY LOOKING AT THE
STAINING AND DO THIS FOR THE
FULL PANEL TO SEE WHAT THE
RESOLUTION IS.
AT HIGH EFFECTER TARGET RATIOS
MOST OF THE CARS ARE ABLE TO
EXPRESS VIRUS REPLICATION AND
THOSE TARGETING THE GP INTERFACE
APPEAR TO MAY BE LESS POTENT BUT
ALL IN ALL ARE POTENT AT TARGET
RATIO.
WE INCREASED THE RATIO OR
DECREASE THE NUMBER OF CARS PER
TARGET WE SEE A DIFFERENT STORY.
HERE MOST OF THE CARS AREN'T
ABLE TO SUPPRESS AND
CORROBORATES SOME DETE -- DATA
WE JUST SHOWED.
WITHIN AN EPITOPE WHAT DICTATES
POTENCY?
THERE'S GREATEST RANGE FROM
THOSE THAT ARE QUITE POTENT AND
WE ASKED THE QUESTION.
IN THIS BINDING ASSAY WE HAVE
NANO PARTICLES AND WASHED THEM
OFF AND CAN IMAGE CELLS TO SEE
HOW MANY ARE LEFT OVER AND THINK
IT'S A FUNCTION OF THE BINDING
AFFINITY.
WE GET A NIGHT SPREAD SUGGESTING
YES THERE'S HETEROGENEITY FOR
THE PHENOTYPE AND IF WE LOOK AT
TWO FUNCTIONAL WITHOUT PUTS OF
THE CARS LOOKING AT THEIR
ABILITY TO KILL TARGET CELLS OR
IL2 ON THE RIGHT AND REGRESS
THOSE AGAINST THE BINDING SCORES
IT APPEARS TO BE A FAIRLY ABOUT
PREDICTOR BETWEEN EP EPITOPES.
HOW TIGHTLY THE CARS BIND
APPEARS.
NOW WE HAVE SOME T CELLS.
WHAT HAPPENS IN VIVO?
WE HAVE A MOUSE MODEL TO TEST
THIS AND TAKE MICE AND
ADOPTIVELY TRANSFER THE T CELL
PRODUCTS AND LOOK AT WHAT
HAPPENS.
THESE ARE SOME OF THE MOST P
POTENT AND DO NOT EXPRESS VIRAL
LOAD.
THEY'LL ESCAPE MUTATIONS AND WE
SEQUENCED THE MICE AND SEE CON
ANTICAL CONTEXT SITES AND THE
LOSS OF GLYCAN AND THE CARS AT
DIFFERENT POSITION SELECT FOR
ESCAPE AND ALL THE GLYCAN AND
THIS SUGGESTS WE CAN DEFINE CAR
T CELL PRODUCT HAVE ORTHOGONAL
ESCAPE PATTERNS AND WE CAN
COMBINE THEM AND GET A DIFFERENT
RESULT.
AND FOR THE OTHER COHORTS WE
PERFORMED A MIXTURE AND HAVE THE
SAME NUMBER OF CAR T CELLS
AGAINST EPITOPES AND WE SEE
CONTROLLERS.
WE'RE ABLE TO FULLY SUPPRESS IN
THE MODEL.
WHAT'S MORE INTERESTING IS WHEN
WE TAKE THEM OUT AND JUST LOOK
AT MICE THAT HAVE HIGH VIREMIA
WE SEE AI LOG REDUCTION IN VIRAL
LOAD RELATIVELY DURABLE
SUGGESTING THE COMBINATION OF
CAR T IS DOING SOMETHING
IMPORTANT WE THINK IS PREVENTING
OR SLOWING THE KINETICS OF
ESCAPE.
TO FORMALLY INTERROGATE THAT WE
SEQUENCED AND LOOKED AT LOSS OF
THE GLYCAN BECAUSE THIS IS
COMMON AMONG THE COHORTS AND
LOOK AT THE RATE AT WHICH THE
GLYCAN IS LOST AND THE MICE THAT
GET THE MIXTURE OF CAR T CELLS
HAVE A LOWER RATE AND THINK IT'S
IMPORTANT BECAUSE THE RATE THAT
IT'S LOSS IS RELATED TO
CUMULATIVE VIRAL LOADS AND IF WE
CAN RESTRICT ESCAPE MAYBE WE CAN
GET BETTER CONTROL.
BECAUSE MIXTURES SEEM TO MAKE A
BIG DIFFERENCE INSTEAD OF MAKING
MIXTURES MAKE A CAR WHERE THEY
EXPRESS THE VARIOUS CAR T CELLS
AND AT LEAST THESE ARE NOT
INFERIOR OR MORE POTENT WHEN WE
LOOK AT THE KILLING ASSAY AND
SECRETE LEVELS SUGGESTING THESE
ARE EQUALLY AS POTENT.
AND SOME OF THIS LED TO US
BELIEVE MAYBE THE CAR T CELLS
HAVE GREATER ANTIGEN SENSITIVITY
AND EXPAND BETTER IN VIVO AND
CAN FORMALLY TEST IN OUR
SYSTEMS.
IN THIS SCENARIO INSTEAD OF
USING ONE TO TRANSDUCE WE USED
THREE WHICH GIVES A COMPLICATED
PRE-TRANSFER PRODUCT BUT AT
LEAST WE HAVE DIFFERENT
FREQUENCIES AND WE TAKE THE
AUTOLOGOUS CAR T CELL PRODUCT
AND TRANSMIT IT TO MICE AND LOOK
RAT WHAT HAPPENS TO CAR
EXPANSION OVER TIME.
THIS SHOWS THE RELATIVE
FREQUENCIES OF THE TOTAL T CELL
COMPARTMENT OF THE MICE IN THE
BOTTOM YOU SEE THE NUMBERS
GIVING THE PRODUCTS AND IT WAS
CLEAR TO SEE WE DO SEE CAR
EXPANSION AND IN THE MICE THAT
DON'T SEE CAR EXPANSION THESE
TWO HERE BECAUSE THEY'RE
CONTROLLING THEIR VIRAL LOAD AND
SUSPECT THERE'S NOT ENOUGH TO
DRIVE PROLIFERATION.
ONE THING THAT'S ENCOURAGING
THAT DUAL AND SPECIFIC T CELLS
MAY HAVE GREATER SENSITIVITY AND
LOOK AT THE FULL CHANGE IN
RELATIVE CAR T CELL TO THE
TRANSFER PRODUCT WE SEE ROBUST
AND PROFOUNDLY ENHANCED
EXPANSION OF THE DUAL AND TRI
SPECIFIC AND WE'LL FOLLOW-UP
WITH MORE IN VIVO EXPERIMENTS
BUT IT'S ENCOURAGING.
TO SUMMARIZE IT APPEARS DISTAL
EPITOPES ARE ASSOCIATED WITH
SUPERIOR CAR POTENCY IT MAY BE
ABOUT EPITOPE ACCESS BUT WE'RE
ACTIVELY IN TER GETTING THAT.
WE KNOW CAR AVIDITY IS MOST
STRONG IN CARS THAT BIND TO THE
SAME EPITOPE.
RESTRICTING HIV ESCAPE SEEMS TO
BE A VIABLE STRATEGY FOR MORE
ENHANCED AN ROBUST SUPPRESSION
OF VIRAL LOAD BUT THERE'S STILL
WORK TO BE DONE.
RIGHT NOW WE'RE TRYING TO MAKE
HIGH ENOUGH NUMBERS OF A PURE
TRI VALENT CAR PRODUCT AND MAY
BE ABLE TO SUSTAIN AND HAVE
BETTER VIRAL LOAD IN VIVO.
I WANTED TO THANK EVERYONE IN MY
LAB BECAUSE EVERYONE'S
CONTRIBUTED TO THE PROJECT AND
HIGHLIGHT THE TALENTED GRADUATE
STUDENT IN MY LAB WHO
SPEARHEADED HIS AND HAS A POSTER
IF YOU WANT TO LOOK AT THAT.
>> OUR NEXT PRESENTATION IS
VIRTUAL IF THE A/V CAN SPOTLIGHT
STEVEN.
>> ARE YOU THERE?
>> I'M HERE.
>> EXCELLENT.
JUST TELL ME WHEN TO ADVANCE.
>> I CAN'T SEE THE SLIDES.
NOW I CAN.
GOOD AFTERNOON, GOOD MORNING,
EVERYONE.
I'M STEVEN MEANLEY AND HIV
PROVIDER SURVEY REVEALS GAP IN
CURE RESEARCH AWARENESS AND BY
THE ENGAGEMENT GROUP AND BOARD.
SO OUR STUDY FOCUSSED ON
UNDERSTANDING THE ATTITUDES OF
CLINICAL PROVIDERS TOWARDS
DISCUSSING HIV CURE RESEARCH
WITH THEIR PATIENTS IN TREATMENT
INTERRUPTIONS OR ATIs.
HOW WILLING ARE THE PROVIDERS TO
DISCUSS HIV CURE STUDY
OPPORTUNITIES WITH THEIR
PATIENTS AND WHAT INFLUENCES
THEIR WILLINGNESS WHEN ATIs ARE
REQUIRED.
TO INVESTIGATE WE ENDED UP
REACHING 64 CLINICAL PROVIDERS
WHO WORK WITH PATIENTS WITH HIV
INCLUSIVE OF MDs MPs AND RNs
ACROSS EIGHT CLINICS IN
PHILADELPHIA AND EACH COMPLETED
A ONE TIME SURVEY.
WE FOCUSSED ON CLINICAL
PROVIDERS AND PRE-COVID SURVEY
OF PATIENTS WITH HIV IN
PHILADELPHIA AND THEY WERE
DEMONSTRATING A HIGH WILL
WILLINGNESS TO PROVIDER AND WE
KNOW PROVIDERS ARE TRUSTED
SOURCES WHEN MAKING HEALTH
DECISIONS AND PROVIDERS WERE OUR
BRIDGE TO PATIENT POPULATIONS.
PROVIDER BUY-IN IS GOING TO BE
IMPORTANT.
WE NEED TO UNDERSTAND WHAT MAKES
PROVIDERS HESITANT OR COMPETENT
ABOUT DISCUSSING STRATEGIES TO
HELP EQUIP THEM WITH THE RIGHT
TOOLS AND STRATEGY TO
COLLABORATE AND SO ULTIMATELY TO
ENSURE MORE PATIENTS HAVE ACCESS
TO THE RESEARCH OPPORTUNITIES.
IN TERMS OF AWARENESS OF HIV
CURE RESEARCH WE FOCUSSED ON THE
SCIENTIFIC APPROACHES AND WE
WERE DESIGNING THE SURVEY WE
LOOKED AT THE WEBSITE WHERE THEY
DESCRIBED THE THIRD ERA OF THIS
AND AND THE APPROACH WAS AROUND
85%.
WE DIDN'T FIND A WHOLE LOT OF
DIFFERENCES IN PROVIDER
CHARACTERISTICS.
WE DIDN'T DO ANY OF THE ANALYSIS
BY SPECIFIC APPROACH BUT AS A
WHOLE.
UNSURPRISINGLY PROVIDERS WHO HAD
SPECIALIZED TRAINING IN HIV
REPORTED MORE FAMILIARITY XAU
COMPARED TO THOSE WITHOUT
SPECIALIZED TRAINING.
OTHER HIGHLIGHTS THERE WAS
ENTHUSIASM FOR OUTREACH AND HIV
CURE RESEARCH.
THEY THEMSELVES WOULD BE OR
LIKELY TO RECOMMEND TO
PARTICIPATE IN HIV RESEARCH.
WHILE THAT'S GREAT NEWS WE FOUND
THIS ENTHUSIASM DROPPED IF HEIs
WERE REQUIRED.
HALF SAID THEY'D BE WILL TO
RECOMMEND AND THE EXPERIENCE WAS
ASSOCIATED WITH PATIENT
WILLINGNESS AND LOOKING AT
WILLINGNESS THE ONLY FACTOR
ASSOCIATED WITH HIV WAS HIGHER
LEVELS OF EXPERIENCE WITH
PATIENT OUTREACH EXPERIENCE.
AND THEN THE LAST WE ASKED
PROVIDERS WHICH RESOURCES TRADE
THEY'D PREFER TO HAVE ON STAYING
UP TO DATE ON PATIENT RESEARCH
OPPORTUNITIES.
WEBINARS AND PUBLICATIONS AND
CONFERENCES AND UPDATES ALL
RECEIVED ENDORSEMENT FROM AT
LEAST 60% OF PROVIDERS.
A
AND THERE'S POTENTIAL FOR
COLLABORATORS IN PHILADELPHIA.
MOST THE PROVIDERS WE SURVEYED
WERE FROM ACADEMIC CLINIC AND IN
BUILDING THE COLLABORATIONS IT
WOULD BE HELPFUL TO LEVERAGE THE
PREFERRED RESOURCES LIKE
WEBINARS, PUBLICATIONS AN EMAIL
SUMMARIES.
IN TERMS OF WHAT THIS RESOURCES
SHOULD ADDRESS WE THINK THAT IT
WILL BE REALLY IMPORTANT TO
ALERT PROVIDERS TO THE DIFFERENT
APPROACHES BEING INVESTIGATED
ESPECIALLY WITHIN JURISDICTIONS
THAT HAVE AN MDC PRESENT LIKE
PHILADELPHIA.
AND IN PROVING AND UNDERSTANDING
THE NECESSITY AND ETHICS AROUND
IMPLEMENTING HIV WITHIN CURES
STUDY AND BUILDING SKILLS AND
SELF EFFICACY FOR OUTREACH AND
PATIENT SUPPORT AS PATIENTS
PARTICIPATE IN STUDIES.
THAT WRAPS UP MY PART OF THE
PANEL.
.
ON BEHALF OF OUR TEAM, THANK YOU
TO OUR FUNDERS AND OUR
PARTICIPANTS AND THANK YOU FOR
YOUR ATTENTION TO THE
PRESENTATION.
>> WE'LL GO TO QUESTIONS IF YOU
CAN SPOT STEVEN AND BOB THEY'LL
UNMUTE AND IF YOU ADDRESS
QUESTIONS ONLINE DO SO IN THE
BEGINNING SO THEY CAN UNMUTE AND
ANSWER.
>> THIS IS A QUESTION FOR BOB.
PHENOMENAL STUDY AS ALWAYS.
FASCINATING DATA.
YOU HAD SHOWN IN THE CONTROLLER
THAT THE VIRAL ASSAY WAS FAIRLY
EASILY ABLE TO GROW OUT VIRUS.
TO CONFIRM THE RESPONSE IS TRULY
CONTRIBUTING TO THE CONTROL I
WOULD ASSUME YOU ADDED
AUTOLOGOUS IGG TO SUPPRESS ALL
OUT GROWTH.
IS THAT SOMETHING YOU FOUND?
>> THANKS FOR THE QUESTION.
SO TO DO THE VIRAL OUTGROWTH
ASSAYS THAT WE HAVE DONE IN THE
PAST WITH AUTOLOGOUS IGG WE NEED
LARGE NUMBER OF CELLS BECAUSE
NEED THREE OUTGROWTH ASSAYS
PARALLEL WITH NO CONTROL IGG OR
PATIENT IGG.
SO THAT'S A DIFFICULT THING TO
DO.
BUT WHAT WE CAN DO IS TAKE THE
ENVELOPES AND IN THIS INDIVIDUAL
IT WAS ACTUALLY LIMITED
DIVERSIFICATION OF THE ENVELOPE
BECAUSE OF THE TREATMENT.
SO WE CAN TAKE THOSE AND DO
PSEUDO VIRUS NEUTRALIZATION
ASSAYS AND ESSENTIALLY LOOKED AT
THREE ENVELOPES THAT DIFFER
SLIGHTLY AND ALL OF THEM ARE
VERY EFFECTIVELY NEUTRALIZED BY
THE PARTICIPANTS IGG.
>> THANKS.
>> I HAVE A COMMENT AND QUESTION
AND RESPONSE TO BOB'S BEAUTIFUL
PRESENTATION.
JUST TO MAKE THE CONNECTION, THE
DATA THAT MARINA SHOWED IN THE
MICE WITH THE REMARKABLE CD 8
CONTROL WAS THE SAME DONOR WITH
REMARKABLE AUTOLOGOUS
NEUTRALIZING ANTIBODIES.
COMMENT IS PERHAPS THEY'RE
SUPPRESSED PUT MIGHT IT FOSTER
AN ENVIRONMENT FOR DEVELOPMENT
OF EFFECTIVE CD 8 AND THE EFFECT
IS IMPACTED IN ANTIBODIES DOING
THIS IN THIS DONOR?
>> YEAH, THANKS.
THIS IS THE SAME PARTICIPANT AND
MARINA MENTION THE SAME CD 8
RESPONSE IN THIS PARTICIPANT.
IT'S INTERESTING TO CONSIDER
ABOUT WHAT IS CRITICAL FOR
CONTROL.
ALL WE CAN SAY IS THAT THE LEVEL
OF NEUTRALIZING ACTIVITY IN THE
PATIENT'S PLASMA IS COMPARABLE
TO ART REGIMENT.
BY ITSELF IS SUFFICIENT TO
MEDIATE CONTROL.
IT'S LIKELY THE CD 8 RESPONSE IS
ALSO INVOLVED AND HOW THEY'RE
RELATED AND WHETHER THE STRONG
NEUTRALIZING ANTIBODY RESPONSE
HAS ANYTHING TO DO ONE THE
STRENGTH OF THE CD RESPONSE IS
A
GOOD QUESTION AND I DON'T HAVE
AN ANSWER FOR THAT BUT TO LOUIS'
EARLIER PRESENTATION SUBJECTING
A STRONG CTL RESPONSE CAN DELAY,
SLIGHTLY REBOUND AND LOWER SET
POINT.
THIS IS A DIFFERENT SITUATION.
THIS IS NO REBOUND.
AND SO THIS IS I THINK THE
REASON WE THINK THE ANTIBODY
RESPONSE IS PROBABLY A MAJOR
FACTOR.
>> IT WOULD BE A GREAT
OPPORTUNITY FOR STUDY ON THE
SAME SAID OF CONTROLLERS MAYBE
WE MAYBE START TO DISENTANGLE
SOME OF THIS.
>> ABSOLUTELY.
>> FREDERICK NATIONAL LAB FOR
MIKE.
REGARDING THE SHIFT IN FREQUENCY
OF HEL PER CELLS BETWEEN THE
VEREMIC AND TREATED INDIVIDUAL
THERE'S TATE IN THE LITERATURE
SUGGESTING IT CAN FUNCTION OVER
TIME AS A DURATION ON ART AND
TRYING TO REBOUND IT SEEMED LIKE
IF I'M REBOUNDING CORRECTLY YOUR
COHORT MAY HAVE BEEN SKEWED
TOWARD A LONGER DURATION AND
SHIFT IN THE FREQUENCY.
>> THE LITERATURE THERE'S QUITE
SMALL AMOUNTS OF DATA IN THE
LITERATURE THAT SAY AN ART THAT
TFH CELLS ARE THE PREDOMINANT
RESERVOIR.
MATIAS HAS A NICE FIGURE IN HIS
PAPERS THAT SHOW THAT OTHERWISE
ALL THE DATA ARE FROM VIREMIC
LYMPH NODES AND THE DONOR ON 20
YEARS OF ART WE BARELY FOUND AN
CELLS WITH THE VIRUS SO WE CAN'T
MAKE MUCH OF A CONCLUSION THERE.
WHAT WE THINK HAPPENS FROM THE
DATA IN VIREMIC CELL THEY'RE
HIGHLY DEPLETED TO THE SAME
POPULATION UNDER ART ALBEIT IN
DIFFERENT INDIVIDUALS BUT HAS A
HIGH BURDEN OF VIRAL RNA.
WE THINK THAT POPULATION IS JUST
BEING KILLED BY THE VIRUS AND
CAN NEVER ESTABLISH A RESERVOIR
ONCE THE PERSON WOULD GO ON ART.
AND SO I THINK WHAT WE'RE SEEING
HERE IS ESSENTIALLY AN EFFECT
THAT THOSE CELLS ALL TFA ARE
HIGHLY SUSCEPTIBLE.
WE KNOW THAT FROM MANY STUDIES.
THEY DON'T FORM A RESERVOIR
BECAUSE THE CELLS DON'T SURVIVE
INFECTION.
>> THANKS.
>> SANDY AND BRAD JUST ASKED THE
SAME QUESTION I WAS GOING TO
ASK.
THE TWO MECHANISMS IF YOU HAVE
POTENT ANABS ON BOARD IT TAKES
MORE TIME FOR T CELLS TO TALK
EFFECT IN THE STEVE DEEKS
INTERCEPT HE SAW IN HIS TRIAL.
AND I WAS STRUCK BY THE FINDING
ABOUT PROVIDERS NOT BEING
WILLING TO REFER TO ATI TRIALS.
AND USE CLINICAL CARE PROVIDERS.
>> BOB, BEAUTIFUL DATA.
YOU SAID RNA LEVELS WERE
SUPPRESSED.
CAN YOU DETECT ANY VIREMIA AT
ALL?
>> THANKS.
NO, WE HAVEN'T DONE SINGLE COPY
ASSAY.
THERE WAS A BLIP OF 29 ASSAYS
YEAR TWO OR SOMETHING BUT IT'S
NOW AND ARE THESE ALL RESEARCH
AND IS THERE INTERPATIENT
VARIATION?
IS IT GREATER FOR CERVICAL LYMPH
NODES BECAUSE WE'VE HAD THAT
EXPERIENCE AND SOMETHING LIKE
XLA IS MORE UNIFORM BUT MAYBE
IT'S NOT THE CASE.
>> THE LYMPH NODES ARE PRIMARILY
TAKEN FOR DIAGNOSTIC PURPOSES.
SO THE REMAINING SAMPLE THAT'S
OBTAINED IS THEN CRYO OBSERVED
FOR RESEARCH PURPOSES.
THE CERVICAL LYMPH NODES TURN
OUT TO BE ME MOST ACCESSIBLE
ONES FOR THE CLINIC WHICH IS
PREDOMINANTLY A RESPIRATORY
DISEASES CLINIC AND LOOKING TO
MAKE SURE THERE'S NOT TB OR
THINGS LIKE THIS.
WE DO HAVE ONE LYMPH NODE SAMPLE
AND ADDED FOUR MORE LYMPH NODES
FROM THE SAME DEMOGRAPHIC COHORT
JUST TO TRY TO DEAL WITH THIS
ISSUE THAT MAYBE A MUCOSAL
DRAINING SITE WILL HAVE
DIFFERENT SUBSETS OF CELLS WHICH
IS VERY REASONABLE TO ASSUME
HIGHER LEVELS OF ACTIVATION AND
MORE MUCOSAL DRAINING SITE.
AND WE HAVE NOT OVERTLY CHANGED
THE CONTRIBUTION OF CD4 HELPER
CELLS TO THE RESERVOIR BY ADDING
THOSE LYMPH NODES.
>> YOU FIND A WAY OF NORMALIZING
FOR THE ACTIVATION THAT MAY BE
PRESENT BY TOTAL NUMBER OF CELLS
OR SOMETHING LIKE THAT?
THAT WOULD BE USEFUL FOR
EVERYBODY TO KNOW WHAT DIVIDE
BY.
>> YEAH.
I WOULD LOVE TO TALK TO YOU
ABOUT THIS AFTERWARD AND ONE OF
THE PROBLEMS OF THE PROCEDURES
WHICH IS 50% EFFICIENT ANYWAY.
WE KNOW WE'RE MISSING CELLS WITH
PROVIRAL READ.
THE OTHER IS SENSITIVE.
>> AND I KNOW THEY'RE SUPPORTING
THE TISSUE REPOSITORY OF THAT
MIKE USED SO IF ANYONE ELSE HAS
AN INTEREST IN LYMPH NODE
BIOPSY.
>> MY QUESTION FOR DANIEL.
THANK YOU FOR THE GREAT TALK.
A TON OF WORK.
BIG CHALLENGE FOR CAR T CELL IS
ANTIGEN BURDEN AND PEOPLE ON
ART.
HAVE YOU DONE WORK LOOKING AT
THE PROXIMAL MIXTURES IN THE
SETTING OF MICE ON ART AND LOW
RAN TI GEN SETTING?
>> NO, WE HAVEN'T BUT
SENSITIVITY IS OUR MAIN
TECHNICAL QUESTIONS WE WANT TO
ANSWER HOW MUCH ENVELOPE DO YOU
HAVE TO HAVE ON THE SURFACE.
IT'S GETTING TO YOUR QUESTION
WHICH IS CRITICAL FOR THE FIELD
AND HOW THIS WOULD WORK IN THE
CLINIC.
ART MODEL SYSTEMS ARE BASED ON
IF WE HAVE A VIREMIC MOUSE CAN
WE FULLY SUPPRESS VIREMIA
WITHOUT ANYTHING ELSE ON BOARD
IS VERY STRINGENT, RIGHT.
BUT IN THE CASE WE'RE TALKING
ABOUT IS WHY WE THINK CERTAIN
CARS ARE MORE POTENT TO SEE LOW
LEVELS OF ANTIGEN AND THOSE MAY
BE MOST SUITED FOR A CLINICAL
SCENARIO TO HAVE ART ON BOARD
WITH MAYBE AN LRA AND GET LOW
LEVELS OF ENVELOPE ON THE
SURFACE.
WE'RE DOING MICRO SCSCOPY TO SE
HOW MUCH IS ON INFECTED CELLS
AND HAVEN'T DONE ANYTHING FOR
INFECTED CELLS.
>> MY QUESTION IS --
>> THANK YOU.
HELLO, MY NAME IS TANIA LOMBO
FOR THE PEDIATRIC RESEARCH
BRANCH AT THE DIVISION OF AIDS
AND HAVE THE PLEASURE TO
INTRODUCE THE MDC TEAM TODAY.
IT STANDS FOR THE PEDIATRIC
ADOLESCENT VIRUS ELIMINATION AND
THE COLLABORATORY IS THE ONLY
ONE FOR THE PEDIATRIC
POPULATIONS LIVING WITH HIV.
IT HAS THE UNIQUENESS OF POOR
RESEARCH FOCUS THE OTHERS DON'T
HAVE.
THAT'S WHY WE'LL SPAEND A LITTLE
BIT MORE TIME SPEND A LITTLE BIT
MORE TIME AND LED BY THIS TEAM
AND THE ANN WELL LEAD THE
OVERVIEW AND INTRODUCTIONS TO
THE TEAM.
THANK YOU.
>> THANK YOU, TANIA.
HOPEFULLY EVERYONE IS AWAKE
AFTER AN EXCITING POSTER
SESSION.
DEBBIE AND I DECIDED IN THE
OVERVIEW TO FOCUS ON SOME OF THE
NEW THINGS THAT HAPPENED IN LAST
YEAR IN PAVE AS WELL AS TO
EXPRESS OUR GRATITUDE TO ALL THE
PEOPLE THAT CONTRIBUTE TO THE
WORK OF PAVE.
SO, JUST TO KIND OF GIVE AN
OVERVIEW FOR THE NEXT I GUESS A
LITTLE BIT OVER AN HOUR.
WE HAVE PRESENTERS SEATED HERE
AS WELL AS A COUPLE VIRTUAL
PRESENTERS AND JESSICA WILL
START US AND WE HAVE KATHERINE
AND PHILLIP VIRTUALLY AND
FOLLOWED BY LESLEY AND MAURICIO
HERE IN THE ROOM.
THE PAVE MISSION TO REMIND
EVERYBODY IS TO USE CUTTING EDGE
SCIENCE TO DEVELOP AND LOOKING
AT RESERVOIRS TO PAVE THE WAY
FOR FUTURE INTERVENTIONAL
STUDIES THROUGH A LIFE TIME OF
REMISSION.
AND WE'RE HOPING TO ADVANCE SOME
OF THE WORK WE'VE BEEN DOING IN
THE INTERVENTIONAL STUDIES IN
THE NOT TOO DISTANT FUTURE.
THESE ARE FOLKS LISTED HERE.
IT'S A BIG GROUP AS TANNIA
MENTIONED WE'RE THE ONLY HIV
PEDIATRIC COLLABORATORY AND TRY
TO BE INCLUSIVE.
WE HAVE NEW MEMBERS LISTED THERE
FROM THE PAST YEAR.
WE'RE GRATEFUL TO THE SUPPORT
AND HELP OF OUR ADVISORY BOARD.
SEE THE FOLKS LISTED AND SHOWN.
LUIS JOINED AS AN AD HOC MEMBER
THIS YEAR AND IS A GREAT
ADDITION.
WE WANT TO GIVE A SHOUT OUT TO
OUR PAVE NIH OFFICERS AND
PARTNERS WHO ATTEND ALL OUR
CALLS AND HAVE BEEN FANTASTIC
CHAMPIONS OF THE WORK WE DO.
WE HAVE SEVERAL INDUSTRY
PARTNERS LISTED HERE AND JUST AS
AN UPDATE FROM THE PAST YEAR WE
ADDED A NEW INDUSTRY PARTNER FOR
THE IG WORK WITH ALL THE
OPTIMIZATION THEY'VE BEEN DOING.
ANOTHER SORT OF THING WE'RE
PROUD OF AND EXCITED TO SUPPORT
IN PAVE ARE A NUMBER OF
DIFFERENT CLINICAL COHORTS AND
BIO BANKS ACROSS THE WORLD
BASICALLY.
WE HAVE SOME THROUGH THE IMPACT
NETWORK.
WE HAVE SOME PREVIOUSLY
SUPPORTED AND NOW THROUGH A
SUPPLEMENT IN THE BEGINNING OF
YEAR FOUR TO SUPPORT CLINICAL
COHORTS IN SOUTH AFRICA,
MOZAMBIQUE AND CONTINUED
COLLABORATION WITH THE EPI-CAL
AND PHACS ENHANCING OUR ABILITY
TO STUDY HIV RESERVOIRS ACROSS
THE AGE SPECTRUM.
AS ONE EXAMPLE OF THE PEDIATRIC
OF THE COHORT AND SOME
ADDITIONAL WORK THAT PAVE HAS
BEEN ABLE TO DO THAT'S BUILDING
OFF OF WORK IN THE IMPACT
NETWORK, DEBBIE PRESENTED SOME
EXCITING WORK AT THE CURRENT
CONFERENCE LAST YEAR ON FOUR
CASES OF REMISSION IN VERY EARLY
TREATED CHILDREN AND SAW THIS
BEAUTIFUL ARTWORK THAT WAS
DEMONSTRATING I GUESS DETECTING
THE PARTICIPANTS.
AND JING WEN WITH ANN HAS A
SUPPLEMENT TO STUDY CNS DELIVERY
AND LOOKING AT THIS IN NON HUMAN
PRY MIGHTS AND TO FURTHER IMAGE
THAT USING IMMUNOPET.
AND WE'VE BEEN PRODUCTIVE IN THE
LAST YEAR AND HAVE A LOT OF
PUBLICATIONS.
FOUR ARE HIGHLIGHTED HERE.
THESE ARE ALL FROM VARIOUS
CLINICAL COHORT AND YOU'LL HEAR
A LITTLE BIT MORE ABOUT THOSE
AND FROM SOME OF THE TALKS
TODAY.
WE'RE LOOKING FORWARD TO NPH
MOVING TO PUBLICATION IN THE
FUTURE.
THE RESEARCH HIGHLIGHTS THE
BULLETS MARK WITH THE THREE
ASTERISKS AND BREAK IT UP BY
NONHUMAN PRIMATE AND HUME
ANRELATED COHORT AND WE'VE BEEN
INVESTIGATING ADOLESCENTS AND
YOUNG ADULTS AND WHO ACQUIRED IT
PERI-NATNATALLY AND THERE'S BEE
COHORT WITH ACQUISITION AND
RESERVOIR SIZE AND WE'VE NOTED
OVER THE PAST FEW YEARS OF PAVE
IN INFANTS EXPOSED AND ACQUIRED
THERE'S NK CELLS SEEN IN
MULTIPLE COHORTS WE'RE
SYNERGIZING AND PUTTING THAT
DATA TOGETHER.
A COUPLE OF OTHER THINGS JUST TO
AND AN ABUNDANCE OF INTACT
VIRUSES AND IDENTIFYING FACTORS
CONTRIBUTING TO VIRAL CONTROL.
A NUMBER OF COMPLEMENTARY
APPROACHES TO SCREEN NOVEL
LATENCY REVERSING AGENTS AND
FURTHER STUDIES USING ORGANOIDS.
IN TERMS OF THE NON-ORGANOID
EXPERIMENTS YOU'LL HEAR ABOUT
BIOLOGICS WITH LONG-TERM VIRAL
EXPRESSION OF THE ART IN HUMAN
AND NONHUMAN PRIMATES AND HAVE
GIVEN IT AT 7 WEEKS OF LIFE AT
RF2 AND 3 AND SIV INFECTED
INFANTS AND HAVE BEEN LEARNING
NUMEROUS LESSONS FROM ANTIBODIES
THAT ARE INITIATION IN INFANT
MACAQUES AND DOING DOSING
STUDIES TO FOCUS IN ON PEDIATRIC
SPECIFIC DOSING FORI AP
INHIBITORS AND AGONISTS AS WELL
AS A NUMBER OF STUDIES WITH
IMMUNOPET OPTIMIZATION.
AND JESSICA WILL TALK WITH US
ABOUT OUR STAKEHOLDER ENGAGEMENT
AND THE EXCITING KNOWLEDGE
DISSEMINATION TO PROVIDERS AND
POLICY MAKERS.
HOPEFULLY YOU ALL HAD A CHANCE
TO VISIT THE POSTERS OF THESE
WONDERFUL ESI SCHOLARSHIP
RECIPIENTS AND WE'RE GRATEFUL
FOR THE SUPPORT OF NIAID TO
BRING THEM HERE.
WITHIN CAME ALL THE WAY FROM
SOUTH AFRICA IF YOU HAVE THE
CHANCE TO TALK TO THEM AT
ANOTHER BREAK, PLEASE DO SO.
AND FINALLY FOR ME WE JUST
WANTED TO THANK OUR PROGRAM
MANAGERS.
WE HAD A FEW WITH US FROM ALMOST
THE BEGINNING OR THE VERY
BEGINNING FROM JOHNS HOPKINS AND
NEW ONES AT EMERY UNIVERSITY AND
TO THANK ALL THE PAVE
INVESTIGATORS AND MEMBERS AS
WELL.
WITH THAT I'LL TURN IT OVER TO
JESSICA.
>> YOU WANTED TO START WITH OUR
CORE LEADERSHIP.
SO WE REACHED A NUMBER OF NEW
STAKEHOLDERS THIS YEAR AND HAVE
BEEN FOCUSSED ON BROADENING OUR
REACH.
MOST OUR REACH IS IN SOUTH
AFRICA BUT HERE YOU SEE SPECIFIC
THINGS.
ONE PARTNER IS THE SOUTHERN
AFRICAN AMERICAN CLINICIAN
SOCIETY.
THEY HELD THEIR FIRST PEDIATRIC
CARE SYMPOSIUM IN 2023 AT THEIR
ANNUAL MEETING.
WE'VE BEEN WORKING WITH THE
INDIVIDUALS TO INCLUDED AND
EXCITED TO SEE THE LIFE TIME
SURVIVORS SHOW UP IN OTHER
PARTNERS' WORK AS WELL.
WE HAD A NUMBER OF VIDEOS AND
ONLINE VIDEO PARTNERSHIPS THAT
POPPED UP AND IN PARTICULAR
TRYING TO WORK WITH SOCIAL MEDIA
INFLUENCERS TO GET THE WORD OUT
AND THINKING ABOUT SPECIFIC
TOPICS THAT MIGHT BE OF INTEREST
LIKE UTIs AND AGING AND YOU SEE
THE PICTURES AND POSTERS AS
WELL.
SO AGAIN SUPPORTING KNOWLEDGE
DISSEMINATION.
WE'VE HAD A NUMBER OF WEBINARS
AND FORTUNATE TO WORK WITH THE
EGPAF AND HAVE BEEN ABLE TO HOLD
WEBINARS WITH EGPAF TO HIGHLIGHT
THE RESULTS IN THE COHORT WORK.
AND THE OTHER THING WE'VE BEEN
TARGETING IS THINKING ABOUT
BROADENING OUR STAKEHOLDER BASE.
NOT JUST WORKING ON THOSE LIVING
WITH HIV BUT FOCUSSING ON
CLINICIANS AND PROVIDERS.
AND WE HAVE AN ARTICLE OUT WITH
NURSING MATTERS THIS YEAR AND
THEY PUT OUT A GLOSSARY AS WELL
AND THINKING HOW TO EXPAND.
OUR GROUP DOES ADVOCACY.
A NUMBER OF GROUPS FOCUSSED ON
THE ADULT POPULATION AND WE
THOUGHT HOW WE CAN BEGIN TO
TARGET AND HOW WE BRING IN
ADMINISTRATIVE HEALTH AND
REGULATORS INTO THE
CONVERSATIONS.
THE CAB CHAIR DR. CARIBE IS
THERE WITH PARTNERS FOCUSSED ON
HOW DO YOU GET THESE INDIVIDUALS
INVOLVED IN TALKING ABOUT MANY
HIV PEER RESEARCH EARLY.
AND FEATURED AT THE CONSENSUS
MEETING AND HAVE BEGUN TO THINK
ABOUT HOW WE COLLABORATE WITH
MAURICIO AND HIS WORK AND
THERE'S BEEN A VOICES PROJECT.
THIS IS WHERE THEY'RE COLLECTING
DATA FROM STAKEHOLDERS WHO ARE
PEOPLE WITH PERINATAL HIV AND
CAREGIVERS AND VARIOUS
REGULATORS AND HERE ON THE
SCREEN IS A QUOTE FROM ONE OF
THE VOICES PROJECTS AND ON THE
SCREEN BEFORE I DIDN'T POINT OUT
BUT IT'S ONE OF THE YOUNG WOMEN
TALKING ABOUT WHY IT'S SO
IMPORTANT.
SO TOWARDS THE FIRST QUARTER OF
NEXT YEAR WE HOPE TO HAVE
SEVERAL INDIVIDUAL VIDEO
PROJECTS THAT FEATURE THESE
VOICES FROM AROUND SUB-SAHARAN
AFRICA.
AND THE COMMUNITY PLAYS AN
IMPORTANT ROLE AND HERE THERE'S
BEEN FANTASTIC WORK AND WE HAVE
A MEMBER TRANSITIONING TO ONE OF
OUR PAVE MEMBERS AND FEATURED ON
A PANELIST AT THE STRATEGIES
MEETING AND PRESENTED ABOUT
PERINATAL HIV AND THE NATIONAL
AIDS COMMISSION IN COUNTRIES AND
SHE HELD THE FIRST SYMPOSIUM IN
MALL -- MALAWI AND JUST REALLY
EXCITED FOR ALL THE WORK THAT
SHE'S BUILT AND CONTINUES TO DO
IN COLLABORATION TRULY WITH THE
ENTIRE CAB.
I KNOW SOME OF YOU HEARD ABOUT
THE EVENT AND WE WON'T PLAY THE
WHOLE VIDEO WE CAB MAKE SURE
PEOPLE HAVE ACCESS BUT SUCH A
NICE PICTURE OF SOME EVENTS THAT
HAPPENED.
DEBBIE IS THERE AND OTHERS.
I'M NOT GOING TO PLAY THE WHOLE
VIDEO BUT IT HAS DANCING AND A
FEATURE OF THE ART EXHIBIT THAT
SHOWS NOT JUST PARTICIPANTS BUT
ALL THE FANTASTIC ART AND IT'S A
FANTASTIC COLLABORATION.
I KNOW OTHER COLLABORATORIES ARE
REALLY BRINGING IN UGANDA SO
IT'S NICE TO SEE EVERYBODY COME
TOGETHER AND WITH THAT -- SEE,
THEY WANT YOU TO SEE THE WHOLE
VIDEO.
SO, KATHERINE.
>> THANK YOU, JESSICA.
IT'S A HARD ACT TO FOLLOW.
I'M PLEASED TO PRESENT ON BEHALF
OF PAVE RESEARCH FOCUS GROUPS 1
AND 3 INVESTIGATORS AND OUR
INVESTIGATORS FOCUS ON
CHARACTERIZING THE EVOLUTION OF
THE HIV RESERVOIR OVER TIME
FOLLOWING VIRAL SUPPRESSION ON
EARLY ART AND THEN SECONDLY
UNDERSTANDING HOST IMMUNE
RESPONSES THAT SHAPE THE
RESERVOIR.
AND TODAY I'M GOING TO SPEAK
ABOUT SOME OF OUR STUDIES THAT
CHARACTERIZE THE EVOLUTION OF
THE RESERVOIR OVER DECADES OF
VIRAL SUPPRESSION AFTER EARLY
ART.
SO HERE'S OUR PLAIN LANGUAGE
SUMMARY.
WHAT IS THEYY -- THE KEY
QUESTION.
WHAT DOES IT LOOK LINE IN
CHILDREN AFTER LONG PERIODS
BEING UNDETECTABLE IN ART.
LOW LEVELS OF INTACT HIV CAPABLE
OF REPLICATION HAVE BEEN
DETECTED IN INDIVIDUALS STUDIED
AFTER DECADES OF BEING
UNDETECTABLE AND THE RESERVOIR
APPEARS TO BE LESS STABLE AND
EASIER FOR THE IMMUNE SYSTEM TO
CLEAR.
AND WHY SHOULD THE COMMUNITY BE
ENCOURAGED ABOUT THE RESULTS OR
PROGRESS PRESENTED?
WE FEEL IMPROVED UNDERSTANDING
OF HIV RESERVOIRS AND THE IMMUNE
RESPONSES THAT SHAPE THEM OVER
TIME CAN INFORM TARGETED
REMISSION STRATEGIES FOR
CHILDREN AND YOUTH LIVING WITH
HIV.
SO THE VERY FIRST EARLY
COMBINATION STUDIES WERE STARTED
29 YEARS AGO AND SINCE THEN
EARLY DIAGNOSIS AND
ANTIRETROVIRAL THERAPIES HAVE
BEEN STANDARD OF CARE.
AND THEY HAVE DEMONSTRATED THAT
EARLY ART CAN MARKEDLY REDUCE
THE SIZE OF THE RESERVOIR AND
REDUCE THE IMMUNE FUNCTION AND
REDUCE MORBIDITY AND MORTALITY.
THE NUMBER OF CHILDREN GROWING
WITH HIV INTO ADULTHOOD OFFERS
THE OPPORTUNITY TO EVALUATE HOW
THE HIV RESERVOIR EVOLVES OVER
TIME.
AND THE HIV COHORT WAS ONE STUDY
AND THEY LOOKED AT NINE
INDIVIDUALS ALL BUT ONE OF WHOM
STARTED ART LESS THAN SIX MONTHS
OF AGE.
THESE CHILDREN EXPERIENCED 8 TO
25 YEARS OF VIRAL SUPPRESSION
AND SINCE THEY ARE WERE OLDER
AND LARGER THEY ARE WERE ABLE TO
DO LEUKOPHORESIS TO OBTAIN LARGE
NUMBER OF CELLS AND 86 MILLION
PBMC WERE INVESTIGATED PER
INDIVIDUAL.
AND WHAT THIS STUDY SHOWED A
LONG STORY SHORT IS THAT IT
SHOWED A SIGNIFICANTLY LOWER
NUMBERS OF INTACT HIV PER
MILLION PBMC AND LOWER PORTIONS
OF INTACT PROVIRUS IN THE
CHILDREN COMPARED TO LEAD
CONTROLLERS OR ADULT INDIVIDUALS
WHO WERE EXPRESSED ON ART.
WE HAVE DONE A STUDY AND WHAT WE
HAD THE OPPORTUNITY TO DO WAS TO
STUDY LONGITUDINAL SAMPLES TAKEN
FROM TWINS LIVING WITH HIV
INFECTION AND THIS FOLLOWED THEM
FROM INITIATION OF
ANTIRETROVIRAL THERAPY THROUGH
28 YEARS OF AGE.
THE FIRST SAMPLES EVALUATE D THY
EXPERIENCED WHY INTRAPARTUM
TRANSMISSION AND ART WAS
INITIATED AT 10 WEEKS OF AGE.
THE P1042 HAD DETECTION AT SIX
MONTHS ON ART AND THE RIGHT
PANEL HAD DETECTION AT 1 1/2
MONTHS AT ART AND THE FIRST
SAMPLES AT EIGHT MONTHS AND 12
YEARS, 20 YEARS AND 28 YEARS AND
THE SAMPLES TAKEN AT 28 YEARS
WERE DONE BY THE LEUKOPHORESIS
TO GIVE US LARGE NUMBERS OF
CELLS.
WHAT WE FOUND IN THE SAMPLES THE
PHORESIS SAMPLES AT 28 YEARS WE
WERE ABLE TO ANALYZE ABOUT A
BILLION CELLS FROM EACH OF THESE
INDIVIDUALS.
WE DID NOT FIND ANY INTACT
SEQUENCES FROM P10 42 AND FOUND
ONE INTACT FROM P1043.
AND WHEN WE LOOKED AT
LONGITUDINAL SAMPLES WE SAW AN
ALMOST FOUR LOG DECREASE IN
INTACT HIV DNA COPY NUMBER AND
THAT WAS COMPARED TO ABOUT A 1-2
LOG DECREASE IN DEFECTIVE HIV
DNA COPY NUMBER FROM EIGHT
MONTHS TO 28 YEARS.
AND THE LEVELS OF INTACT
PROVIRUSES WERE 80 FOLD LOWER
THAN THOSE DETECTED IN
CONTROLLERS AND 6700 TIMES LOWER
THAN THOSE MEASURED IN ADULT
INDIVIDUALS WHO INITIATED ART
DURING ESTABLISHED INFECTION.
AND THEN IN TERMS OF DETECTIVE,
WE ALSO SAW A TODAY CREASE BUT
NOT QUITE SO MUCH AND THE
DEFECTIVE PROVIRAL LEVELS WERE
24 FOLD LESS THAN THOSE MEASURED
IN ELITE CONTROLLERS AND 160
FOLD LESS THAN ADULTS THAT
INITIATED ART WITH THE
ESTABLISHED INFECTION.
DEF
THIS SHOWED INTACT IN
INDIVIDUALS AND THE INTACT
PROVIRAL COPY NUMBER DECREASED
FROM 8,000 FOLD AND BY CONTRACT
DEFECTIVE PROVIRAL LEVELS
DECREASED BY 64 FOLD OVER THAT
TIME PERIOD.
IN THIS LEVEL IT DECREASED BY
13,000 FOLD AND PROVIRUS LEVELS
DECREASED BY 98 FOLD AND BODY
WEIGHT INCREASED AGAIN THE TWO
SAMPLING TIME POINTS INCREASED
ONLY 8 TO 9 TIMES AND SINCE THE
NUMBER OF LYMPHOCYTES ARE
ROUGHLY PROPORTIONAL TO BODY
WEIGHT WE DO NOT THINK THE
DECREASE IN PROVIRAL LEVELS IS
SIMPLY DUE TO A DILUTIONAL
EFFECT.
WE THEN WENT WE USED 298 MILLION
CD8 CELLS IN AN ASSAY AND DID
NOT OBSERVE -- WERE NOT ABLE TO
RECOVER VIRUS FROM EITHER
INDIVIDUAL.
WE BEGAN TO LOOK AT THE PROVIRAL
LANDSCAPE OVER TIME AND WHILE WE
DETECTED EXPANSION OF INTACT AT
THE FIRST TIME POINT IN BOTH
INDIVIDU
INDIVIDUALS AND WE DID NOT
DETECT EXPANSION OR ANY INTACT
CLONE SUBSEQUENTLY.
IN P1043 WE DETECTED A SMALL
CLONE AT 12 YEARS BUT NOT
SUBSEQUENTLY.
AND OVER ALL WE OBSERVED LIMITED
GENOMIC DIVERSITY AND SAW
LIMITED EXPANSION OF DEFECTIVE
CLONES AT THE LATER TIME
PERIODS.
BOTH INDIVIDUALS WERE ANTIBODY
NEGATIVE AT THE TIME OF TESTING
AT 28 YEARS.
THEY PRE-ART WERE ABLE TO DETECT
CD 8 T CELL AND MAP CD 8 T CELL
RESPONSES IN THEM AFTER VIRAL
EXPRESSION AND RESPONSES WERE
NOT DETECTED BY DIRECT EX VIVO
OR INTRACELLULAR CYTOKINE SAERS
BUT WERE ABLE TO DETECT CD4 AND
CD 8 T CELL RESPONSES WHICH
INDICATE THEY HAD MEMORY.
THEY PRESERVE MEMORY RESPONSES
THROUGH 28 YEARS OF AGE AND TO
GAG AND A POOL OF PEP SIDES.
-- PEPTIDES.
IN SUMMARY AFTER DECADES OF
EXPRESSIVE ART WE OBSERVED MARK
REDUCTION IN THE MANY DNA AND
THE SEQUENCES WERE EXTREMELY LOW
AND DECREASED OF INTACT PROVIRAL
VIRUSES EXCEEDED THAT OF
DEFECTIVE SEQUENCES.
WE DID NOT SEE EXPANSION OF
CLONES WITH INTACT FOR VIRAL
SEQUENCES OVER TIME AND THE DATA
SUGGEST A LONG-TERM STABILITY
AND INCREASED IMMUNOLOGIC
PROVIRUSES STARTING IN EARLY
INFANCY.
AGAIN WE WERE ABLE TO DETECT
PERSISTENT HIV SPECIFIC T CELL
RESPONSES AND WE ALLUDED TO ARE
NOT GOING TO FOCUS ON INNATE AND
K CELL FUNCTIONS AND LESLEY WILL
BE SPEAKING MORE ABOUT THAT IN
HER PRESENTATION LATER ON THIS
AFTERNOON.
SO BEFORE I CLOSE I'D LIKE TO
ACKNOWLEDGE STUDY PARTICIPANTS
AND THEIR FAMILIES AS WELL AS
MEMBERS OF THE LABS.
MY LAB, THE POWER LAB, LESLEY DE
ARMAS AND MEMBERS OF THE
CHILDREN'S HOSPITAL AND I THINK
I'LL STOP THERE.
THANK YOU VERY MUCH.
>> THANK YOU, KATHERINE.
PHILLIP ARE YOU ON?
>> YES.
MY TALK IS EARLY LIFE IMMUNE SEX
DIFFERENCES IN HIV CURE
POTENTIAL.
AND THESE ARE THE PEOPLE
INVOLVED IN THE HUMAN STUDIES
COMPONENT OF RESEARCH FOCUS 2
AND 3 AND I'LL BE SPEAKING ABOUT
PART OF THIS WORK IN THE NEXT
MINUTES.
IN TERMS OF PLAIN LANGUAGE
SUMMARY.
THE KEY QUESTION CAN CURE
REMISSION BE ACHIEVED BY EARLY
ANTIRETROVIRAL SYSTEM AND DO
IMMUNE SEX DIFFERENCES PLAY A
ROLE?
SINCE SEX DIFFERENCES DO AFFECT
OUTCOME, THIS WORK ADVANCES OUR
FIELD BECAUSE IT MEANS THAT
IMPLIES SEX SPECIFIC
INTERVENTIONS MAY BE IMPORTANT
TO ADVANCE CURE POTENTIAL AMONG
CHILDREN.
WHY SHOULD THE COMMUNITY BE
ENCOURAGED ABOUT THE RESULTS?
WE IDENTIFIED FIVE CHILDREN IN
THE STUDY WHO REMAINED
UNDETECTABLE FOR UNSCHEDULED
ANTIRETROVIRAL INTERRUPTION AND
EIGHT CHILDREN TO A PROSPECTIVE
STUDY AND FOUR CHILDREN REMAINED
UNDETECTABLE MORE THAN A YEAR
AND OTHER FINDINGS JUST MENTION
ED INDICATE THE POTENTIAL TO
CURE CHILDREN AND BROADLY
NEUTRALIZING ANTIBODIES CAN BE
IMPLEMENTED.
THE STUDY I'LL TALK ABOUT WAS
THE BABY CURE PROJECT WE STARTED
10 YEARS AGO IN NORTHERN EASTERN
PART OF AFRICA FOR THE STUDY
SITES SHOWN HERE.
THERE'S 320 CHILD PAIRS
FOLLOWING IN UTERO TRANSMISSION
AND THEY ARE HAVE REMISSION
FOLLOWING EARLY ANTIRETROVIRAL
AND LOOK AT THE CURE REMISSION
IN THESE INSTANCES.
THE SEX DIFFERENCES CAME FROM
SOME OF THE TWINS WE STUDIES IN
THE COHORT AND WE'VE COME ACROSS
FIVE SEX DISCORDANT TWINS WHERE
ONE HAS BECOME INFECTED AND IT
WAS THE FEMALE AND THE
HISTORICAL CODE IN MALAWI AND
THE SUSCEPTIBILITY TO
ACQUISITION IS GONE TO RECENT
MATERNAL INFECTION LOOKING AT
MOTHERS WHO HAD CONVERTED BEFORE
PREGNANCY THERE WAS NO
TRANSMISSION TO MALES AND
FEMALES BUT IN THE MOTHERS WHO
CONVERTED DURING PREGNANCY AND
THE MECHANISM CAME FROM ADULT
STUDIES WHICH HAS SHOWN THE
VIRUS IS TRANSMITTED TO ADULTS
FOR RESIST JANS AND THIS BECOMES
AND MOTHERS RECENTLY INFECTED IN
THE COURSE OF PREGNANCY WILL BE
MORE LIKELY TO HARBOR RESISTANT
VIRUSES AND HYPOTHESIZED LIKE
FEMALE CHILDREN AND FEMALE
ADULTS THE FEMALE FETUSES MAKE
STRONGER RESPONSES THAN MALE
COUNTERPARTS.
FRALE FETUSES ARE MORE RESISTANT
AND THE RESISTANT VIRUSES ARE
HARBORED BY THE MALES AND FEMALE
FETUS ARE MORE SUSCEPTIBLE THAN
MALES IN UTERO.
WE LOOKED AT SOME SEX DISCORDANT
TWINS BORN TO HIV-NEGATIVE
MOTHERS AND THE STIMULATED GENES
WAS HIGHER IN CD4 CELLS IN
FEMALES VERSUS MALES AND THE
LEVEL OF ACTIVATION IN THESE
WERE HIGHER AMONG THE FEMALES
AND THE LEVELS WERE CORRELATED
WITH THE ABILITY TO INFECT THE
CELLS IN VITRO WITH HIV.
I WANTED TO SHOW YOU THE VIRUSES
TRANSMITTED TO MALES AND FEMALES
IN THE STUDIES WE'VE BEEN DOING
AND SHOWING ON THE LEFT PANEL
VIRUSES TEND TO BE HIGHER
CAPACITY ON THE Y AXIS AND
INTERFERON SENSITIVE ON THE X
AXIS.
AND BY CONTRACT THE VIRUS IS
TRANSMITTED TO FEMALES BUT THE
MAJORITY OF THEM ARE IN THE LOW
RIGHT HAND QUADRANT WITH LOWER
CAPACITY.
THESE ARE VIRUSES THAT YOU
RARELY SEE TRANSMITTED TO MALES.
AND MOST TRANSMITTED TO FEMALES
WERE TRANSMITTED IN THE SETTING
AND HAVING SEEN MOST THE VIRUS
WERE TRANSMITTED TO MALES ARE
TRANSMITTED AND FIT VIRUSES WE
HAVE ALSO OBSERVED TYPICAL MALES
WITH VIREMIA FOR SUBSTANTIAL
PERIODS UP TO 10 MONTHS
FOLLOWING UNSCHEDULED
INTERRUPTION.
WE WERE ABLE TO CHARACTERIZE
VIRUSES IN THE TYPICAL CASES AND
SHOWED THEY'RE UNUSUALLY LOW IN
TERMS OF THE CAPACITY IN MAIL
INFANTS AND SUGGESTING IN MALES
THE VIRUSES TRANSMITTED AND
SUSTAI
SUSTAINED FOLLOWING CHILD BIRTH
ARE AT CAPACITY IN MALES.
AND WE HAVE ALSO INITIATED SOME
PERSPECTIVE STUDIES IN EIGHT OF
THE CHILDREN AND FOUR WE HEARD
FROM THE PAST FOUR WEEKS AND
OTHERS REMAINED IN REMISSION AND
OTHERS OCCURRED AT 18 MONTHS.
I'D LIKE TO FINISH BY
ACKNOWLEDGING ALL THE PEOPLE WHO
PARTICIPATED IN THE STUDY
PARTICULARLY THE STUDY
PARTICIPANTS AND CLINICAL TEAMS.
THE TEAM THAT PARTICIPATED IN
THE WORK I DESCRIBED AND NAMED
ON THE LEFT AND ALSO THE FUNDERS
THE PAVE IN PARTICULAR AND
STUDIES THEMSELVES WERE FUNDED
BY WELCOME TRUST.
THANK YOU.
>> I'M HERE TO PRESENT DATA ON
BEHALF OF THE INVESTIGATORS NOT
JUST MYSELF.
THIS IS THE RF4 TEAM OF HUMAN
STUDIES.
I'M NOT PRESENTING WORK FULLY
FROM THIS GROUP BUT FOCUSSING ON
THE IMMUNOLOGICAL ASSAYS WE ARE
DEVELOPING.
WE HAVE TWO MAIN QUESTIONS.
ONE IS CAN WE MEASURE NATURAL
KILLER WITH PERINATAL HIV WITH
SAMPLE AVAILABILITY AND WHICH
CHARACTERISTIC ASSOCIATES WITH
VIRUS CONTROL IN THE POPULATION.
HOW DOES THIS WORK ADVANCE THE
FIELD AND MK CELLS ARE
COMPONENTS OF THE INNATE
IMMUNITY AND PLAY A ROLE IN THE
ANTIVIRAL IMMUNITY AND DRIVING
NK KILLING POTENTIAL IN HIV AND
INFANTS.
THE COMMUNITY SHOULD BE
ENCOURAGED ABOUT RESULTS AND
PROGRESS BECAUSE IT CAN DELAY TO
THE DECAY OF THE HIV RESERVOIR
AND THERE'S STUDIES IN PEDIATRIC
POPULATIONS WHERE WE HAVE
LIMITED VOLUMES.
AND THE FINDINGS ARE EXPECTED TO
PROVIDE INSIGHT TO STRATEGIES TO
BOOST HIV IMMUNITY TO TARGET THE
RESERVOIR.
OUR WORK IN DOING IMMUNOLOGIC
PROFILING AND PERINATAL HIV IS
TRYING TO DETERMINE WHAT
RESPONSE IS PRESENTS AND/OR
EFFECTIVE PRE AND POST OUR
INITIATION.
WE HAVE THREE MAIN TOOLS WE USED
TO INTERROGATE AT THE SYSTEM.
THE FIRST IS DOING HIGH
DIMENSIONAL FLOW CYTOMETRY AND
SECOND IS FUNCTIONAL STUDIES TO
DETERMINE HOW DIFFERENT CELL
TYPES RESPOND TO EXTERNAL
STIMULI AND VIRAL INFECTION AND
mRNA SEQ TO LOOK AT THE
UNBIASSED VIEW OF THE LANDSCAPE.
THE GOAL IS TO OPTIMIZE SAMPLES
SPARING ASSAYS TO LOOK AT
COHORTS IN CLINICAL TRIAL AND
LOOKING AT NK CELL DATA WE
GENERATED USING THE THREE
APPROACHES.
THAT WILL COME FROM TWO
DIFFERENT AND THE STUDY COMES
FROM BOTSWANA LED BY ROBERT
SHAPIRO A TREATMENT WITH ATI IN
CHILDREN THREE TO FOUR YEARS OF
AGE.
THEY WERE TREATED VERY EARLY AND
WERE ON CONTINUOUS ART SIX WEEKS
WERE UNDETECTED AND IN M
MOZAMBIQUE WE WERE FOLLOWING A
COHORT AND THIS STUDY ENROLLED
CHILDREN STUDIED EARLY AT 1 TO 2
MONTHS AND ZERO NEGATIVE INFANTS
WITH IN UTERO AND WE HAD
LONGITUDINAL SAMPLING AND
CLINICAL FOLLOW-UP.
FIRST TO SHOW THE NK FINDINGS
AND THIS IS WORK DONE SHOWING
THE SCHEMATIC OF THE STUDY SO
CHILDREN WERE GIVEN ART AND THE
ATI WENT ON 24 WEEKS UNLESS
VIRUS LOADS WENT ABOVE THE
THRESHOLD AND WE HAD 14
REBOUNDERS AND 11 CONTROLLERS SO
THOSE WHO MAINTAINED EXPRESSION
FOR THE 24 WEEKS.
AND MELANIE PERFORMED FLOWCY
CYTOMETRY.
AND WE'RE FOCUSSING MAINLY ON
TWO POPULATIONS THE CD 56
POSITIVE AND 16 NEGATIVE AND
CYTOKINE PRODUCING CELLS AND THE
MAJOR CYTOTOXIC POPULATION AND
LOOKED AT FREQUENCIES AND THERE
WAS NO SIGNIFICANT DIFFERENCE IN
SUBSET FREQUENCIES.
THEY FOUND SIGNIFICANT DIFFERENT
IN CELLS EXPRESSING THE NK
RECEPTOR TO THE L1 AT THE TIME
OF OUR INITIATION IN BOTH THE NK
CELL SUBSETS THAT WERE MEASURED
AND THE LEVEL OF THIS INHIBITORY
MARKER EXPRESSION ON THE CD 56
DIM CELLS HAD A NEGATIVE
CORRELATION WITH WEEKS TO VIRAL
REBOUND SUBJECTING THE SUBSET
WAS NOT ABLE TO CONTROL HIV
REBOUND.
SO NK CELLS EXPRESS A DIVERSE
ARRAY OF RECEPTORS THAT DICTATE
LIGATION WITH DIFFERENT LIGANDS
ON CD4 AND OTHER CELLS DICTATE
THEIR FUNCTION WHETHER
INHIBITORY OR ACTIVATING AND THE
MAJOR LIGAND FOR KIR IN THE 2DL1
IS HLA-C.
AND THE EXPRESSION VARIES AMONG
TYPE AND MELANIE EVALUATED THE
DIFFERENT TYPE AND FOUND LOW
EXPRESSION ALLO TYPES WERE IN
CONTROLLERS AND HIGH TYPES WERE
ENRICHED IN REBOUNDERS AND
IMMUNE MEDIATED CONTROLLERS MAY
PLAY A ROLE IN VIRAL REBOUND.
SWITCHING OVER TO THE MANY TARA
COHORT.
IT DOES NOT HAVE AN
INTERVENTION.
WE'RE LOOKING AT IMMUNOLOGIC
PROFILES IN RELATION TO AGE,
VIRAL STATUS AND ART INITIATION.
AND WE'VE BEEN INTERESTED IN NK
CELLS BECAUSE WE FOUND SOME
DIFFERENCES IN SUBSET
FREQUENCIES AND PHENOTYPES
BETWEEN AGI AND AGU.
SO WE WANTED TO DEVELOP AN ASSAY
TO LOOK AT NK FUNCTION BECAUSE
WE THINK IT'S CRITICALLY
IMPORTANT IN INFANCY.
SO WE DID THIS IN NINE ATU AND
17ATI AND LOOKED AT ENTRY ONE TO
TWO MONTHS OF ABLE AND AGAIN AT
12 MONTHS OF ABLE AND THE ASSAY
WE START WITH PDMC AND PURIFY NK
AND CO-CULL CURE WITH TARGET
CELLS AND PERFORM A 25 COLOR
FLOW CYTOMETRY ASSAY TO LOOK AT
CYTOTOXIC FUNCTION AND CYTOKINES
AND MARKERS OF DEEP GRANULATION.
THE TARGET CELLS ARE CEM AND KR
THAT ARE NK RESISTANT AND MHC1
DEFICIENT AND NK SENSITIVE AND
HUD 78 A CELL LINE THAT STABLY
INFECTED WITH SF 2 AND HIV 1
STRAIN AND THESE CELLS ACTED AS
OUR HIV INFECTED TARGET CELLS.
SO THE PRIMARY READOUT OF THIS
ASSAY IS JUST LOOKING AT
SPECIFIC KILLING OF DIFFERENT
CELL LINES.
WE DID NOT SEE A DIFFERENCE
BETWEEN AGI AND AGU SUBJECTING
NO INHERENT DEFECT IN AGI AS FAR
AS TARGETING CELLS BUT THERE'S
NO RELATION WITH VIRUS LOAD AT
THE 1-2 MONTH TIME POINT.
HOWEVER AT 12 MONTHS WE DID SEE
A NEGATIVE CORRELATION BETWEEN
SPECIFIC KILLING OF 78 CELLS AND
PLASMA VIRUS LOAD AND ONLY 16
WERE SUPPRESSED AND DID NOT SEE
ASSOCIATION WITH THE KILLING OF
K562 SUBJECTING FRO LONGED
VIREMIA PLAY MEDIATE KILLING OF
HIV INFECTED CELLS.
WE WANTED TO LOOK AT WHICH NK
FEATURES CORRELATED WITH TARGETS
OF KILLING.
SO WE DID MIXED EFFECTS MODELS
WHERE WE LOOKED AT ALL THE
DIFFERENT PHENOTYPIC MARK FROM
THE FLOW CYTOMETRY DATA SET AND
SAW A POSITIVE IMPACT OF CD 25
EXPRESSION AND THE CELLS AND
SMALL POSITIVE EFFECT OF LIGAND
RECEPTOR ON THE CD 56 STEM
CELLS.
TO SEE HOW THIS LOOKS IN
CORRELATION PLOTS WE SEE A NICE
LINEAR POSITIVE RELATIONSHIP
BETWEEN LIGAND EXPRESSED AFTER
THE CULTURE AND FREQUENCY OF
CELLS THAT WERE KILLED.
WHEN YOU LOOK AT THE CELLS YOU
SEE CD 107A AND THE MARKER
HAVING A STRONG ASSOCIATION AS
WELL AS THE BETA AND INHIBITORY
MARKER AT EXPECTED AS A NEGATIVE
IMPACT.
SO WE'RE SEEING THE NK
PHENOTYPES DIVERGE IN
CO-CULTURES DEPENDING ON THE
CELLULAR TARGET.
SO THE EMERGING DATA SO FAR
SUGGESTING NK RECEPTOR AND CD4
LIGANDS MAY PLAY A ROLE IN
CONTROL AND WENT TO A DATA SET
OF 12ATI AT THE PRE-ART TIME
POINT AND HAD GENE AND PROTEIN
EXPRESSION FROM 130 ANTIBODIES
TO LABEL PDMC.
WE WERE ABLE TO SEPARATE OUT
SOME OF OUR HIGH VIRUS VERSUS
LOW VIRUS LOAD IN INFANTS AND
THE MAP IS SHOWING CLUSTERS.
WE IDENTIFIED SEVEN CD4 T CELL
CLUSTERS WHICH WERE ANNOTATED AS
NAIVE OR BASED ON A REFERENCE
DATABASE.
WE ALSO IDENTIFIED TREGS AND TH2
POPULATION DUE TO EXPRESSION AND
THE CLUSTERS YOU CAN SEE THEY
ALMOST ACROSS THE BOARD HAVE UP
REGULATED HLA EXPRESSION AND AN
ANTIBODY THAT DOES NOT
DISTINGUISH BETWEEN HLA AND CR C
AND HIGH UPREGULATION AND THE
FOUR CLUSTERS IDENTIFIED WHICH
WERE ABLE TO ANNOTATE USING CD
56 AND CD 66 EXPRESSION.
WE SEE UP REGULATION OF SOME
INHIBITORY TEARS HIGH VERSE US
AND SEE UPREGULATION IN THE
INDIVIDUALS WITH HIGHER VIREMIA.
SO THESE RESULTS THE IMMUNE
PROFILES CONSISTENT WITH REDUCED
NK FUNCTION IN THE PRESENCE OF
HIGHER AMOUNT.
AND THE INHIBITORY MARKERS ARE
ASSOCIATED WITH THE REBOUND AND
DIDN'T SHOW THE KG2A AND
PROLONGED VIREMIA IN INFANTS
IMPAIRS KILLING OF HIV INFECTED
CELL LINES BUT THE K562 CELLS
AND THERE'S A MECHANISM USING
LIGANDS RATHER THAN A CD107A
DEGRANLATION AND FAS EXPRESSION
WAS INCREASED WITH HIGHER VIRUS
LOAD AND SINGLE CELL RNA SEQ ARE
FEASIBLE FOR SAMPLES AND XASHG
TWO SMALL GROUP SUCH AS HIGH LOW
RESERVOIR OR CLINICAL RESPONSES.
AND I'LL THANK INDIVIDUALS FROM
THE LAB WHERE I WORK AND THOSE
WHO DID THE EXPERIENCE AND THE
ANALYSIS AND MOZAMBIQUE TOOK
CARE OF THE INFANTS FOLLOWING
AND WE APPRECIATE THEIR
CONTRIBUTIONS AND THEN MATHIAS
AND MELANIE FOR THE DETE AND THE
WHOLE TEAM.
IT WAS A REALLY INTERESTING
SITE.
>> GOOD AFTERNOON.
I'M MAURICIO MARTIN AND TALK
ABOUT THE AAV-VECT DELIV DELIVE
INFANT MACAQUES.
OUR GROUP IS SHOWN HERE.
I WANT TO MENTION THE STUDY I'LL
BE TALKING ABOUT NOW WERE
CONDUCTED UNDER THE SEPARATION
OF DR. HAYWOOD AND SO MY PLAIN
LANGUAGE SUMMARY SO THE KEY
QUESTION WE ADDRESSED IS CAN
EARLY ART PLUS GENE THERAPY WITH
HIV-1B NABS ACHIEVE DURABLE
ART-FREE CONTROL IN VIREMIA IN
INFANT MACAQUES AND LED TO
PRODUCTION OF THE MONKEY'S OWN
CELLS WITHOUT REPEATED
ADMINISTRATION.
AFTER STOMP ANTIRETROVIRAL ABOUT
80% OF THE TREATED MACAQUES
MAINTAINED BNAB LEVELS THAT
PREVENTED VIRUS REBOUND.
WE SHOULD BE EXCITED ABOUT THIS
FOR TWO REASONS.
THE BNAB VECTORS IN GENERAL ARE
HIGHLY STABLE AND ACROSS
MULTIPLE TEMPERATURES AND
CONDITIONS MAKING THEM
ESPECIALLY USEFUL FOR DEPLOYMENT
TO REMOTE AREAS WHERE A COLD
CHAIN CANNOT BE RELIABLE.
AND THE ADMINISTRATIONS CAN BE
DONE WITHOUT ADVANCED MEDICAL
TRAINING OR FACILITIES WHICH
HAVE A PLUS AND BNAB EXPRESSION
IN INFANCY PLAY PREVENT LATER
INFECTION IN LIFE.
WE LIKE THIS APPROACH OF USED
AAV TO DELIVER B NABS BECAUSE IT
CAN PERSIST IN THE NUCLEUS OF
THE CELL WHERE IT CAN MAINTAIN
PERSISTENT EXPRESSION FOR THE
CELL LIVE AND WE'RE INTERESTED
IN TARGETING THESE VECTORS TO
SKELETAL MUSCLE CELLS BECAUSE OF
THE LIFE LONG SPAN.
OUR VISION FOR THE APPROACH IS A
ONE-TIME INTRAMUSCULAR WOULD
PROVIDE PROTECTION AND I'LL TALK
ABOUT THREE STUDIES CONDUCTED OR
IN THE PROCESS OF CONDUCTING OUR
.
OUR 2022 STUDY WE ORALLY
INFECTED THE INFANT MACAQUES AND
ONE WEEK LATER WE START THE
ANIMALS ON ANTIRETROVIRAL AND
THEY RECEIVED INTRAMUSCULAR
VERSIONS AND THEN AFTER WEEK 30
POST INFECTION WE STOPPED ART
AND ASSESSED VIRUS REBOUND AND
WE SAW A SIGNIFICANT DELAY OR
PREVENTION AS SHOWN IN THE TOP
RIGHT.
SO I'M GOING TO TO BE TALKING
ABOUT THE STUDY FIRST.
THIS IS SHOWING ONE PANEL FOR
ANIMAL AND I'VE SHOWN THIS
BEFORE BUT UPDATE WHAT WE'VE
DONE RECENTLY.
WE SAW TWO ANIMALS IN THE 2022
COHORT EXPERIENCED VIRUS REBOUND
AND INTERESTED IN CHARACTERIZING
THE SEQUENCE OF THEIR ENVELOPE
OF VIRUSES AT THE TIME OF
REBOUND.
I ALSO WANT TO POINT OUT THAT WE
IN THE LAST YEAR CONDUCTED
RECEPTOR BLOCKADE IN VIVO AND
SAW REBOUND CONSISTENT WITH THE
DECREASE IN PLASMA
CONSIDERATIONS OF THE TWO
BIOLOGICS AND AS THE EXPRESSION
OF THE BIOLOGICS CAME BACK,
VIROLOGIC CONTROL WAS
RE-ESTABLISHED AND PEOPLE
GETTING THE MOLECULES AND AS FOR
THE ENVELOPE SEQUENCES WE
IDENTIFIED WE'RE STILL
CHARACTERIZING THEM BUT
IDENTIFIED A NUMBER OF
POLYMORPHISMS SHARED BETWEEN THE
ANIMALS ANALYZED AND THE SHARED
POLYMORPHISM INVOLVED A DELETION
IN V4 AND THE SEQUENCING DATA
WAS GENERATED BY DR. BRANDON
HEEL'S GROUP AND HAVE BEEN
CHARACTERIZING THE PROVIRAL
LOADS IN THE ANIMALS AND
COMPARED THEM BETWEEN THE
MACAQUES AND THE CONTROL GROUP
OVER 30 YEARS OF ANTIRETROVIRAL
IN STUDY AND HADN'T SEEN A
DIFFERENCE PROBABLY BECAUSE OF
THE PERIOD OF ART BEING TOO
SHORT.
AND WE'RE PLANNING TO ASSESS
THIS IN THE FUTURE WITH LONGER
ART PERIODS.
SO IN THE 2023 STUDY WE HAVE
STOPPED ART AND SO FAR WE HAVE
SEEN STABLE CONTROL IN FUF OF
THE SIX ANIMALS THAT WERE
ENROLLED.
WE HAVE A 7th MACAQUE ENROLLED
BUT THIS IS ENCOURAGING BECAUSE
IT SHOWS CONTROL CAN BE
MAINTAINED BY THIS APPROACH IN
ANIMALS THAT WERE TREATED WITH
ART AND AAV VECTORS AT 7 AND 21
DAYS POST INFECTION.
SO WE'RE IN THE PROCESS OF
CONDUCTING THIS STUDY WHERE
WE'RE ANALYZING THE ADVOCACY OF
THE VERSIONS OF THE 10-1074 AND
HAVE EXPRESSION OF AT LEAST ONE
OF THE MOLECULES WITH THE ATI
PLANNED FOR FEBRUARY OF 2025.
TO SUMMARIZE WHAT I SAID SO FAR,
WE SEE PREVENTION OR DELAY OF
VIRUS REBOUND AND ABOUT 80% OF
MACAQUES AT WEEK ONE OR THREE
POST SHIV INFECTION AND WE KNOW
THAT MANY WE SHOULD TRANSIENTLY
DEPLETE THE CIRCULATION THROUGH
BLOCKADE AND THE VIRUS COMES
BACK.
WE ARE -- WE JUST DID TWO CD 8
DEPLETIONS IN THE 2023 COHORT
AND WE HAVEN'T SEEN THE VIRUS
COME BACK YET SO THIS IS I THINK
STRENGTHENS THE CONCLUSION IT'S
REALLY THE EXPRESSION OF THE TWO
MOLECULES MAINTAINING CONTROL
AND SO FAR WE HAVE THE 2024
COHORT WHERE WE ARE AWAITING
ANXIOUSLY FOR THE AGI IN 2025.
AS FOR PLANS FOR THE FUTURE
WE'RE IN ACTIVE CONVERSATIONS
ABOUT TRANSLATING TO A CLINICAL
TRIAL AND CONDUCT THE MONKEY
STUDY TO ASSESS WHETHER A
COMBINATION OF VERSIONS OF
101074 AND IT CAN ACHIEVE THE
LEVEL OF CONTROL WE HAVE SEEN IN
STUDIES I JUST DESCRIBED.
WE ARE CONTEMPLATING USING THIS
DUAL COMBO FOR THE CLINICAL
TRIAL AND FOR THIS PARTICULAR
STUDY WE ARE COLLABORATING WITH
DR. KATIE BARR AROUND A SHIV
THEY DEVELOPED AND HAVE INFECTED
THREE INFANT MACAQUES WITH THE
VIRUS.
WE'RE LOOKING AT REPLICATION AND
HOW THE VIRUSES RESPOND TO
ANTIRETROVIRAL THERAPY AND ATI
AND THAT'S SHOWN IN THE GRAPH.
AND YES, THIS IS MY
ACKNOWLEDGEMENT SLIDE.
THANK YOU VERY MUCH FOR ALL
PARTIES INVOLVED ESPECIALLY THE
MEMBERS OF MY LAB AND THE ASSAYS
AND MY COLLEAGUES AT THE CENTER
WHO DID THE ANIMAL WORK.
THANK YOU.
>> THANK YOU.
WE'RE OPEN FOR QUESTIONS.
WE HAVE 10, 12 MINUTES.
HERE WE GO.
>> QUESTION FOR MAURICIO.
THE QUESTION OF THE ANTIBODY IN
SPITE OF EXPRESSION DID YOU SEE
IMPACT OF THE CD 8 RESPONSE IN
THE EXPRESSION IN THE
CONJUNCTION WITH THE RESPONSE OF
THE ANTIBODY?
>> SO, I THINK I COVERED THIS A
BIT TOO QUICKLY.
WHAT I MEANT TO SAY IS THAT IN
TWO OF THE 2023 COHORT ANIMALS
WE TREATED THEM WITH CD 8 ALPHA
DEPLETING ANTIBODY TO SEE IF THE
VIRUS WOULD COME BACK AND
HAVEN'T SEEN ANY VIRUS OF LAST
WEEK.
WE HAVE NOT CHARACTERIZED CD 8 T
CELL RESPONSES IN THE ANIMALS
BECAUSE OF THEIR LIMITED BLOOD
AVAILABILITY AND WILL CONTINUE
MONITORING PLASMA FOR VIRAL
LOADS IN THE NEXT WEEKS TO SEE
IF THAT TRIGGERS ANY REPLICATION
BUT IT HASN'T SO FAR.
>> AT THE HIV MEETING IT WAS
SHOWED USING TWO NEUTRALIZING
ANTIBODIES AND THREE DAYS AFTER
INFANT MACAQUE INFECTION HE CAN
CURE SIX OUT OF SIX ANIMALS, I
BELIEVE.
YOU ARE PLANNING TO ADD
ANIRANOMAB IN THE FUTURE?
>> WE'RE PLANNING TO COLLABORATE
WITH HIS GROUP.
AND IN THE NEAR FUTURE LET'S PUT
IT THIS WAY.
>> I HAVE A QUESTION FOR
KATHERINE.
>> IT SEEMS LIKE I WAS THINKING
DO YOU THINK PROLIVE TIF
CAPACITY OF THE SPECIFIC CD 8 T
CELLS WOULD YOU CALL IT HIGH OR
CALL IT LOW?
WHERE WOULD YOU PUT THAT IN THE
AND THE QUESTION ABOUT THE
ADULTS AND THE RESERVOIRS AND
TRYING TO THINK ABOUT HOW THESE
PROLIVE -- PROLIFERATIVE
CAPACITY IS HOW STRONG IS?
>> WE MEASURED IN THE ASSAYS THE
PROLIFERATIVE CAPACITY TO GAG
AND THE OTHER WAS COMPARABLE TO
WHAT WE HAVE SEEN IN ADULTS WITH
HIV.
IT WAS A GOOD RESPONSE AND I
THINK IT MAKES SENSE GIVEN WE
WERE ABLE TO DETECT HIV SPECIFIC
AND MAP HIV SPECIFIC CD 8 T
CELLS PRIOR TO INITIATION OF ART
IN THE KIDS.
>> I WAS THINKING THAT IF THE
QUALITY SEEMS PARTICULARLY GOOD
LIKE IF THE PROLIFERATIVE
RESPONSES SEEMED GOOD MIGHT THEY
RESPOND WELL TO THERAPEUTIC
VACCINES?
AND I WAS CURIOUS WHAT THE
THOUGHTS ARE --
>> THEY HAVE RESPONDED NORMALLY
TO ROUTINE CHILDHOOD VACCINES
AND WE WOULD EXPECT THAT THEY
ARE WOULD PROBABLY HAVE A
SECONDARY RESPONSE OR A GOOD
RESPONSE TO A THERAPEUTIC
VACCINE.
>> LIKE A T CELL BASED
THERAPEUTIC VACCINE.
>> EXACTLY.
>> ARE THERE PLANS FOR THAT?
>> WELL, WE PLANNED FOR IT
SEVERAL TIMES OVER THE YEARS.
THE AVAILABILITY OF VACCINE IS
LOOKING FOR A VACCINE WE THINK
IS CAPABLE OF GENERATING THAT
TYPE OF T CELL RESPONSE.
>> I CAN ADD TO THAT RACHEL.
SO THE IMPACT WE HAVE STUDIED
2039 IN DEVELOPMENT ALREADY
AGAIN TO REVIEW AND WILL INCLUDE
THE VACCINE WITH BNAB IN 5 TO
12-YEAR-OLDS TREATED IN INFANCY.
>> MAURICIO, I NOTICED IN YOUR
CONTROL ANIMALS, IF I HAD IT
RIGHT THE VIREMIA COMES DOWN IN
THE CONTROL.
IT LOOKS LIKE YOU HAVE A HIGHER
RATE OF CONTROL THAN WHAT
PHILLIP WAS SAYING FOR EXAMPLE
IN THE HUMAN PEDIATRIC COHORT
AND MAKES THE LIKE LIKE THE
MODEL HAS MORE CONTROL AND THE
ANTIBODIES MAY HAVE MORE EFFECT
THAN THE SHIV.
CAN YOU COMMENT?
>> THAT'S SOMETHING WE SEE OFTEN
WITH SHIV WHICH REMAINS AN
IMPERFECT MODEL.
WE HOPE AND PREVENTION OF
REBOUND AND IF YOU LOOK AT THE
PARAMETER ALONE AND IN THE
PRESENCE OF THE EXPRESSED
BIOLOGIC WE SEE A SIGNIFICANT
DELAY IN TERMS OF VIREMIA POST
ATI WE HAVE FEW ANIMALS, ONLY
ONE ACTUALLY OF THE 16 SO FAR
THAT IS VIREMIC AFTER ATI.
SO WE CONSENT REALLY MAKE THE
COMPARISON BETWEEN POST ATI
VIRAL LOADS.
>> THE DYNAMICS YOU WERE
DESCRIBING WERE TIGHTLY LINKED
TO THE WINDOW OF INFECTION OF
THE MOTHER SO THEREFORE IS IT
RIGHT TO THEN INTERPRET THE
ACUTE INFECTION WITH THE VIRUS
IS MORE INTERFERON RESISTANT IS
KEY DYNAMIC TO THE PREFERENTIAL
OF THE FEMALES OR THE INTRINSIC
BASELINE OF THE RESPONSE ON THE
FEMALE IS THE DRIVER MEANS IF
YOU HAVE WOMEN THAT TRANSMIT
PASSED THE WINDOW OF ACUTE
INFECTION YOU DON'T SEE THAT
DIFFERENTIAL ANYMORE?
>> THAT'S RIGHT.
I THINK THE DATA IS SAYING IN
THAT SETTING THERE'S NO
DIFFERENCE IN TERMS OF MALES TO
FEMALES AND THAT SETTING IS
INTERFERON SENSITIVE.
IF IT'S TRANSMITTED TO MALES IS
RARELY INTERFERON RESISTANT AND
THE REASON WE THINK IS TO DO
WITH THE IMPACT IT HAS ON
REPLICATE CAPACITY.
THERE'S A NEGATIVE CORRELATION
BETWEEN CAPACITY AND YOU NEED A
REASONABLY FIT VIRUS TO GET
ACROSS THE TRANSMISSION BO
BOTTLENECK IN MALES AND THE FEW
THAT SUCCEEDED ACROSS THOSE ARE
THE ONES WHERE WE SEE AVIREMIA.
>> THAT'S WHAT I WAS GOING TO
FOLLOW UP NEXT.
YOU THINK THE CLINICAL
SIGNIFICANCE DURING THE
SUSCEPTIBILITY TO INFECTION OR
WILL HAVE A LONG-TERM IMPACT
HAVING BEEN TRANSMITTED AT THAT
POINT TO WHAT ACTUALLY HAPPENS
SUBSEQUENT?
>> I THINK IT'S BOTH.
AND IF YOU HAVE A SENSITIVE
VIRUS YOU'RE LIKELIHOOD OF
CONTROLLING THAT VIRUS IS GOING
TO BE HIGHER.
>> THANK YOU AND NOW WE'LL MOVE
TO THE NEXT.
>> SO THUR -- THANK YOU EVERYONE
I'M IN THE TARGETED
INTERVENTIONS PROGRAM IN THE
SCIENCES PROGRAM IN THE DIVISION
OF AIDS AND OVERSEE THE CRISPR
FOR CURE COLLABORATORY.
SOMETIMES YOU SEE IT AS C4C.
THE CODIRECTORS ARE KAMALL
KHALILI AND TRICIA BURDO AT
RUTGERS UNIVERSITY.
PLEASE GO AHEAD.
>> THANK YOU VERY MUCH FOR
INCLUDING US IN THIS EVENT.
YOU MAY KNOW THE CRISPR FOR CURE
IS ONE OF THE LATEST PROGRAMS TO
BECOME PART OF THE MARTIN
DELANEY CURE PROGRAM FOR HIV.
IT'S VERY FOCUSSED IN TERMS OF
THE GOAL AND THE GOAL OF THE
MARTIN CRISPR FOR CURE IS TO USE
MOLECULAR GENETIC APPROACH FOR
REMOVAL OF THE INTEGRATED COPY
OF THE HIV FROM THE HOST
CHROMOSOME AND FOR YELL LAR
GENES WHOSE PRODUCTS ARE
IMPORTANT FOR VIRAL REPLICATION.
IT'S THE PLAIN LANGUAGE SUMMARY
IS SHOWN HERE.
IT'S A QUESTION WE'RE ASKING CAN
BE LIMITED HIV DNA INTEGRATED IN
CELL CHROMOSOME WHICH HAPPENS
SOON AFTER THE VIRAL INFECTION
AND ALSO THAT CHROMOSOME SERVES
AS THE TEMPLATE FOR PRODUCTION
OF THE NEW VIRAL PROJECT.
SO THEN KEY FINDINGS AND TAKE
HOME MESSAGE FROM THE STUDIES WE
HAVE DONE SO FAR IS THAT CRISPR
GENE EDITING TECHNIQUES HAVE
DEMONSTRATED I BELIEVE TO ENTIRE
VIRAL GENOME AT THE CELL CULTURE
AND DESIGNATED SEGMENT DICTATED
BY THE GUIDE RNAs FROM THE HOST
GENOME YOU SEE IN THE CARTOON ON
THE RIGHT SIDE.
AND WHY THIS IS IMPORTANT IS
IT'S IMPORTANT BECAUSE THIS
METHOD OFFERS A POTENTIAL
GENETIC BASED TECHNIQUE FOR THE
REPLICATION COMPETENT VIRUS
WHICH IS TO MY KNOWLEDGE IS A
CRITICAL STEP TOWARD THE YOU'RE.
AND CURE OF HIV.
WHY SHOULD WE BE EXCITED ABOUT
THIS?
BECAUSE BY DOING THAT THE
INDIVIDUALS ARE BASICALLY
INFECTED BY HIV AND DON'T NEED
TO TAKE ANY ART AND THE
COMORBIDITY ASSOCIATED WITH THE
PRESENCE OF THE VIRUS OR THE
TREATMENT STRATEGIES WILL
DISAPPEAR AND WON'T BE STIGMA
ASSOCIATED WITH THE VIRUS
PRESENCE.
THIS DEMONSTRATED HAS WE
STARTED.
IN 2015 WE USED CRISPR
TECHNOLOGY FOR THE VIRUS FROM
THE CELL CULTURE SYSTEM AND SOON
WE REALIZED IT'S ACTUALLY
POSSIBLE TO REMOVE ENTIRE VIRAL
FROM THE LGR IN THE CELL CULTURE
MODEL AND THIS WAS REPRODUCED
SEVERAL TIMES IN A NUMBER OF THE
CELLS AND PUBLISHED IN THE 2014.
AND CRISPR CAN PROBABLY BE USED
FOR THE CURE OF HIV AND THEN
LATER ON IN 2020 WE MOVED WITH
THE LARGER ANIMAL NONHUMAN
PRIMATE AND WE USED THIS AS THE
TOOL FOR THE INFECTION WHICH IS
AN ABOUT MODEL FOR HIV AS YOU
ALL KNOW AND THEN WE SHOWED THAT
THE APPLICATION OF THE CRISPR
FOR TARGETED REGION RESULTS IN
THE REMOVAL OF THE AREA FROM THE
HOST GENOME.
AND THEN DELIVERY WE USED FOR
THAT WAS AAV9.
AND THEN WE REALIZED IT'S
BROADLY DISTRIBUTED TO ALMOST
EVERY CELLS AND THE RESULT OF
THE DISTRIBUTION WAS EXCISION
FROM THE GENE IN THE DNA AND
THIS OBSERVATION FOR WHY THE
CONVINCING EVIDENCE FOR THE
CRISPR EXCISION BY THERAPEUTIC
TEAM TO GO TO NIH AND GET FDA
APPROVAL AND THE CLINICAL
TRIALS.
WE'RE NOT GOING TO TALK ABOUT
THE OUTCOME OF THAT STUDY
BECAUSE IT'S OUT OF THE SCOPE OF
THE MARTIN DELANEY.
SO WE WHAT YOU'LL HEAR ARE
STUDIES USED IN THE NONHUMAN
PRIMATE FOR THE EDITING OF THE
VIRAL GENOME SO THEN AT THAT
POINT WE ALSO FORMED A TEAM.
A TEAM THAT INCLUDED OTHER SMALL
GROUP AS YOU PROBABLY NOTICED
COMPARED TO OTHER DELANEYS AND
THEN AND CASE WESTERN UNIVERSITY
AND UCLA AND EXCISION WAS OUR
CORPORATE PARTNER AND A TEAM
MOVED TO RUTGERS AND INITIATED
THE PROGRAM AT THE RUTGERS
INSTITUTE OF TRANSLATIONAL
RESEARCH AND THE BIO QUAL WHICH
IS BASICALLY A FACILITY FOR
NONHUMAN PRIMATE STUDIES CLOSE
TO HERE AND ALSO WASHINGTON
UNIVERSITY SCHOOL OF MEDICINE
ST. LOUIS.
SO AS YOU SEE IT'S VERY FOCUSSED
BEFORE PEOPLE GOT TOGETHER AND
CARRIED ON THE STUDIES WITH THE
CENTRAL THEME AND THE GOAL IS
ERADICATION OF THE INTEGRATED
COPY OF THE VIRUS.
THE PROGRAM IS COMPRISED OF OF
THE RESEARCH FOCUS AND WE HAVE
AN RF1 FOCUSSING ON THE
UTILIZING BAR CODE SIB KINDLY
PROVIDED AND FOR THE STUDIES AND
USING ROLLOVER EPIGENETIC IN THE
CHRIS PER -- CRISPR EDITING
APPROACH AND THE LATENT TISSUE
RESERVOIR AND IMAGING ANALYSIS
AND THEN LOOKING AT THE VIRUS
AND BASIC SCIENCE AT THE CORE OF
OUR GROUP AND RF2 IS INVOLVING
THE LOOKING AT THE NUMBER OF
AREAS WHICH CAN HELP TO
UNDERSTAND THE VIRUS HOST
INTERACTION AND POTENTIAL EFFECT
OF THE VIRUS IN ELIMINATION OF
THE VIRUS AND THE AREA OF FROM
FOR OUR TEAM.
ON ONE HAND WE USED THE
INFLAMMASOME CONTROLLING SIV
DISEASE PROGRESSION AND CRISPR
MODIFICATION IN MONKEY MODEL AND
ALSO IMMUNOMODULATION AND THE
EXPANDING OR ACTIVATING NK CELLS
AND FINALLY THE CRISPR EDITING
IN THE NONHUMAN PRIMATE AND WE
HAVE DONE THAT PREVIOUSLY IN THE
MICE BUT TRYING TO SEE WHAT
RESPONSE WE GET IF WE CAN PAIR
THE ELIMINATION OF THE
ACTIVATION OF SIV IN CONTEXT OF
THE VIRUS.
AND THE RF3 IS BASICALLY
TECHNICAL ASPECT OF THE CRISPR
WHICH INCLUDES THE TOOLS
CREATING FOR THE GENETIC
ANALYSIS AND THEN LOOKING AT THE
OFF TARGET, ON TARGET SEQUENCE
AND THE FOCUS IS TO CREATE THE
BEST AND EFFICIENT PRODUCTS.
THE PRODUCTS IS THE CAS 9 RNAs.
THE TOOL FOR THE DELIVERY WE
USED WAS AT THAT STAGE WAS AAV9
AND THEN OBVIOUSLY MODIFYING
THAT DELIVERY SYSTEM AS WE MOVE
ALONG.
SO THIS IS JUST AN EXAMPLE OF
THE FEW PUBLICATION THAT WE HAD
OVER THE LAST YEAR OR TWO.
AND THEN AS YOU SEE NOTICE IT'S
COMING FROM THE DIFFERENT
LABORATORIES PARTICIPATING IN
THIS GROUP FROM RF1, RF2 AND
RF3.
SO, WITH THAT NOTE I'M GOING TO
START BASICALLY TALKING ABOUT
SOME OF THE OBSERVATION WE HAD
IN THE RF3 FOCUSSING ON THE
CREATION OF THE CRISPR GUIDE RNA
FOR THE VIRUS IN THE NONHUMAN
PRIMATE.
THE VIRUS THAT WE USED WAS
BASICALLY A BAR CODED AS I
MENTIONED PROVIDED TO US BY OUR
COLLEAGUE HERE AND THEN THIS
LOOKS AT BASICALLY FOR THE
INCORPORATION OF THE BAR CODED
SEQUENCE AND THEN AFTER THE
SERIES OF THE ASSAYS THEY ARE
DEVELOPED SOME IN THE LABORATORY
WE IDENTIFIED THE REGIONS WITHIN
THE OUTSIDE OF THE BAR CODED
SEQUENCE WHICH CAN BE USED FOR
INACTIVATION OF THE VIRUS.
OBVIOUSLY WE DIDN'T WANT TO USE
THE AREA IN THIS PART OF THE
GENE YOU HAVE TO FOCUS ON THE
PART AWAY FROM THE BAR CODED
SEQUENCE.
SO THE RESULT FROM THE ANALYSIS
USING THE SINGLE RNA FIRST TO
IDENTIFY THE REGION WHICH CAN
SERVE FOR DEVELOPMENT OF THE
GUIDE RNA FOR ELIMB FLATION --'
ELIMINATION AND DIDN'T RESULT
THE ENTIRE ELIMINATION AND
REMOVED THE SEQUENCE AND
IDENTIFIED THE REGION OF THE
VIRUS AS YOU SEE HERE .
ALSO NEXT TO THAT WAS THESE
REGIONS AND THESE TWO BASED ON
THE ANALYSIS SHOWED INEFFICIENCY
FOR THE EDITING OF THE GENE.
AND HERE YOU CAN SEE EVEN IN THE
PAIR WHEN WE PAIRED THEM THESE
TWO YOU SEE GOOD EFFICIENCY OF
T
THE REMOVAL OF THE DNA.
SO WE MOVE ON WITH THE TARGETS
IN THE STUDIES.
SO THIS SHOWS SOME OF THE
RESULTS IN TERMS OF THE RESULT
FROM THE LOSS OF SIGNAL DDPCR
SHOWED 31% OF THE EXCISION OF
THE REMOVAL AND USED ANOTHER
CANDIDATE AND THE NUMBER WAS
MUCH LESS THAN THAT SO WE'LL
MOVE FORWARD.
AND THEN THE NEXT ONE WAS
FOCUSSING ON THE VIRAL VECTOR.
FOR VIRAL VECTOR AS WE NOTED THE
SMALLER IS BETTER TO USE AS
SMALL AS YOU COULD BECAUSE OF
THE PACKAGE INEFFICIENCIES.
SO FOR THAT WE IDENTIFIED THE
ELONGATION AND ONE ALPHA
PROMOTER WHICH BASICALLY SHOWS
GOOD EFFICIENCY FOR THE
EXPLANATION OF THE CAS 9.
THIS WAS CREATED AND YOU SEE ON
THE RIGHT AND THEN VALIDATED IN
THE CELLS FOR THE EXPIRATION OF
THE CAS 9 RNA AND THE VIRAL DNA.
THERE'S NO SIGNIFICANT
DIFFERENCE IN THE PRODUCTION OF
THE CAS 9 ELONGATION ONE ALPHA
PROMOTER VERSUS GENETIC PROMOTER
WE USUALLY USE OR BEING USED BY
MANY OTHER PEOPLE.
AND THEN WE SHOWED BY SEQUENCING
THE EX CISION OF THE SEGMENTS.
THIS CONSTRUCT WAS PACKAGED IN
THE NORTH CAROLINA VECTOR
LABORATORIES.
AND THEN WE BASICALLY START THE
EXPERIMENT IN THE MONKEY
INJECTION DONE IN THE FACILITY
AND FACILITY WHICH IS EXISTING
HERE AND THEN SOME DETAILS ARE
SHOWING HERE.
SO WHAT I'M GOING TO DO IS SHOW
YOU THE BRIEF SUMMARY OF THE
VAST NUMBER OF THE DATA THAT WE
OBTAINED.
I WON'T GO THROUGH THAT BUT SHOW
YOU ONE OF THEM IS THE LYMPH
NODE AND IF YOU LOOK HERE YOU
SEE THE CRISPR AND ATI AND NE
NECROPSY IS HIGH.
IT SHOWED THE REBOUND HAPPENED.
BUT THE THING HERE IS WE WERE
HOPING WE DIDN'T SEE BUT WE DID
SEE THE REBOUND AND WE NOTICED
WHEN WE LOOK AT THE EXPIRATION
OF THE CAS 9 IT WAS EXPERIENCING
BECAUSE THAT WAS A CONTROL.
THERE WAS AN EXPERIMENTAL SET
DID NOT GET GUIDE RNAs WHICH IS
BLINDED.
YOU DON'T SEE THAT ONE.
WE ALSO FOUND IN THE SUMMARY IN
THE LIVE LYMPH NODE NINE COPIES
OF THE DAP SEQUENCE DETECTED IN
10,000 CELLS.
AND IN THE SPLEEN THIS WAS THREE
COPIES PER 10,000 CELLS AND WE
DIDN'T TAKE ANYTHING AND IN THE
BRAIN ALSO UNDETECTABLE.
AND WHEN YOU LOOKING AT THE AAV
THE NUMBER SHOWS THE AAV WAS
BROADLY DISTRIBUTED ALMOST IN
THE LIVER AND THEN ALSO TO A
LESSER DEGREE IN THE SPLEEN AND
IN THE LYMPH NODE AND MORE IN
THE BRAIN.
AND THEN WHEN YOU THE ASSAY IT
SHOWED THE RESULT SHOWS
DETECTION OF THE FRAGMENTS
ACROSS THE BOARD ESPECIALLY IN
ONE OF THE ANIMALS WE USED.
IN THE BRAIN AND SPLEEN AND
LIVER SHOWS DETECTION OF THE
FRAGMENTS OF THE VIRUS.
THIS SLIDE ILLUSTRATES THE GUIDE
RNA EXCISION HERE AND HERE AND
AS YOU SEE IN THE NUMBER OF ONE
OF THE ANIMALS YOU SEE THEIR
EXCISION IN THE BRAIN AND SPLEEN
IN SOME WASN'T COMPLETE EXCISION
BASICALLY IN THE MUTATIONS.
INTERESTINGLY WE SAW PRECISE
EXCISION OF THE VIRAL DNA YOU
SEE IN THE CASE AND IN THIS
CASE.
SO THIS EXPERIMENT DEMONSTRATED
THE VIRAL DNA CAN BE EXCISED
FROM THE BROAD RANGE OF TISSUE
THAT SERVES AS THE VIRAL
RESERVOIR BUT THE PROBLEM WAS
DELIVERY WHICH I THINK DELIVERY
WAS NOT AS STRONG.
WE DECIDED TO CHANGE THE
DELIVERY SYSTEM AND THAT'S AN
EXPERIMENT IN PROCESS.
SO I DON'T HAVE A CLEAR UPDATE
FOR YOU TO SHOW IN THE MONKEYS.
BUT I'M GOING TO JUST SHOW YOU
THE DELIVERY SYSTEM WAS BASED ON
SOMETHING PUBLISHED RECENTLY AND
THAT IS A PSEUDO LENTI VIRUS
WHICH BASICALLY CARRIES THE
GUIDE RNAs AND CAS 9 PROTEINS
AND ALSO WE CAN EXPRESS THE GFP
IN THAT AND THE DETECTION THEN
WE INTRODUCED THE CONSTRUCT IN
THE CD4 AND CD 8 POSITIVE AND
NEGATIVE AND SHOWED IT CLEARLY
SHOWS YOU CAN INCREASE THE
NUMBER OF THE CD4 POSITIVE
RECEIVED AND THAT THE DELIVERY
SYSTEM.
SO PERHAPS THIS IS SOMETHING
THAT WE HAVE TO MOVE FORWARD.
WE UNDERSTAND THAT WE SHOWED
THAT THE BRAIN AND LYMPH NODE IN
PARTICULAR RECEIVE THE GUIDE RNA
AND VIRAL GENOME BUT WE NEED TO
GET THE DELIVERY SYSTEM AND HERE
WE HAVE PROBABLY SOMETHING THAT
WE HAVE TO MOVE FORWARD ON AND
WORK ON IT.
I THINK I'LL STOP AT THIS STAGE.
>> I'M TRISH BURDO ONE OF THE
CORE LEADERS AND INVESTIGATOR
AND TALKING ABOUT THE WORK IN
THE NONHUMAN PRIMATES AND TRYING
TO GIVE A FEEL NOR ASSAYS WE DO
FOR RF1 AND OTHER THINGS IN RF2.
THIS IS OUR PLAIN LANGUAGE
SUMMARY.
AFTER USING CRISPR TARGETING DNA
IN MONKEYS WHEN WE STOP
ANTIRETROVIRAL?
AND HOW DO WE MEASURE THE
RESERVOIR WITH CUTTING EDGE
METHODS ADAPTED TO SIV.
KEY FINDINGS AND TAKEAWAY
MESSAGES.
SO CRISPR WAS DELIVERED TO THE
SIV INFECT ED MONKEYS AND SHOW
DATA FROM STUDIES WE'VE SHOWN
THERE WAS NO SIDE EFFECTS AND
STUDIES ARE UNDERWAY TO
DETERMINE IF CRISPR AFFECTED THE
VIRAL RESERVOIR AND METHODS
ADAPTED FOR SIV.
WE'RE TRYING TO DETERMINE HOW IT
WORKS AND IF IT'S SUCCESSFUL IN
REDUCING VIRAL COURSE.
WHY SHOULD WE BE EXCITED?
IF WE CAN SHOW CRISPR CAN REDUCE
SIV IN MONKEYS IT COULD
POTENTIALLY DO THE SAME IN
PEOPLE.
OVER ALL THE MORE WE UNDERSTAND
ABOUT HOW TREATMENTS AFFECT THE
VIRAL RESERVOIR THE BETTER WE
CAN DESIGN AND TARGET FUTURE
STRATEGIES.
RF1 IN OUR LABORATORY RAN BY
DR. ZACH.
NEITHER COULD BE HERE BUT WE
HAVE JOCELYN KIM DESCRIBING HER
WORK IN RF1.
SO ONE OF THE MAIN AIMS OF RF1
IS TO USE BAR CODED VIRUS TO
LOOK AT THE SOURCES OF VIRAL
REBOUND AND THE MECHANISMS OF
VIRAL PERSISTENCE.
WE'RE USING TWO MODELS IN OUR
COLLABORATORY.
THE HUMANIZED MOUSE MODEL AND
THE NONHUMAN PRIMATES AND WE'LL
BE TALKING TODAY MOSTLY ABOUT
THE NONHUMAN PRIMATE WORK.
SO AS KAMALL MENTIONED WE'RE
USING THE BAR CODED VIRUS AND
THERE IS THE BAR CODE HAS THREE
34 NUCLEOTIDE BANDWIDTH AND 10
RANDOM NUCLEOTIDES AND WHAT
WE'RE USING THIS FOR IN THE
STUDIES IS TO DEEP SEQUENCE THE
RESERVOIR TO LOOK AT PLASMA
VIRUS TISSUE RNA AND DNA AND
ANALYSIS OF OUR HIV THAT
REBOUNDS.
SO WE'RE USING THIS ASSAY TO SEE
CRISPR IS SUCCESSFULLY
DECREASING THE RF2.
AND THEY ARE HAVE BEEN DESIGN
ING THE RNA BAR CODE ASSAYS THAT
WORK IN SIV.
BRIEFLY THEY USED A UMI THAT IS
ATTACHED TO A VIRAL BAR CODE
THAT ENABLES THEM TO LOOK AT THE
NUCLEOTIDE AT EACH POSITION AND
SEQUENCE AND GET RID OF PCR OR
DEEP SEQUENCING.
SO THEY'RE USING THE TECHNOLOGY.
>> A/V GUIDES WE JUST NEED THE
SLIDES.
NOT THE SPEAKER.
>> SCARY, EVERYONE SEES ME.
THANK YOU.
THE WORK WORKS WITH BRANDON
KEEL'S GROUP LOOKING AT THE
ASSAYS AND DID A COMPARISON
ACROSS THE LABS AND THE OTHER
METHOD WAS FROM DR. KIM'S LAB
AND SHOWED THERE'S A STRONG
CORRELATION IN THE RESULTS.
SO THE NUMBER OF BAR CODES, THE
DIVERSITY OF THE BAR CODE AND
ABUNDANCE OF THE BAR CODE WAS
ALL VERY CORRELATIVE AND SO WE
BELIEVE THIS SHOWED THE ASSAY IS
ROBUST AND SO WE'RE GOING TO
UTILIZE THE VIRUS TECHNOLOGY IN
OUR STUDIES.
TO GIVE AN EXAMPLE OF THIS.
ON THE LEFT SIDE WE HAVE AN
MRALZ AND ACUTE INFECTION DAY 14
AFTER INFECTION AND SEE A NUMBER
OF BAR CODES.
ON THE LEFT-HAND SIDE IS THE
FREQUENCY.
YOU SEE A LOT OF ABUNDANT BAR
CODE AND VARIANTS ATHET BOTTOM
OF THE FREQUENCY AND MINOR
VARIANCES AND SAMPLE ON THE
RIGHT SIDE ARE THE ART SAMPLE SO
THIS IS ONE WEEK AFTER ART.
AND YOU SEE A DECREASE IN THE
BAR CODES AND THEN YOU ALSO SEE
A DROP OUT OF THE MINOR BAR CODE
AND SO REALLY THAT'S BECAUSE
ONCE YOU START ANTIRETROVIRAL IN
THE CIRCULATION THOSE ARE THE
ONES THAT ARE MOST EASILY
EXPELLED.
WE'RE USING THIS ASSAY THROUGH
OUR STUDY AND WILL BE USING IT
TO LOOK AT DIVERSITY IN CHRIS
PER.
-- CRISPR.
THIS BRINGS US TO OUR DESIGN AND
BRIEFLY WE USED SAB MAC 239M AND
14 DAYS LATER STARTED
ANTIRETROVIRAL AND MONITORED THE
ANIMALS DRAWING BLOOD AND
LOOKING AT VIRAL LOADS AND SO
YOU CAN SEE VIRAL LOADS WERE
INCREASED AND AS SOON AS WE PUT
THE ANIMALS WE STARTED TO
DECREASE AND WE DIDN'T DO OUR
CRISPR STUDIES UNTIL ALL THE
ANIMALS IN THE STUDY WERE WELL
SUPPRESSED.
IN THIS EXPERIMENTAL SET UP WE
HAVE 10 ANIMALS.
FIVE RECEIVED THE AAV9 THAT
INCLUDE THE PSI AND GAG.
WE DID THIS BECAUSE WE WANTED TO
KEEP THE BAR CODE AND OTHER
STRATEGIES IN THE PAST HAVE BEEN
LTR AND GAG AND WE THOUGHT IT
WILL ONLY BE RETRAINED IN ONE
AND WANTED TO DO SOMETHING S
SIMPLE AND RETAIN THE BAR CODE
AND THE OTHERS RETAINED PLASMA
AND IT'S A CONTROL AND WHAT
HAPPENS AFTER CRISPR TREATMENT
DURING THE ATI?
CAN CRISPR DECREASE THE VIRAL
DIVERSITY OR BAR CODE?
DOES IT MAKE AN IMPACT ON THE
RESERVOIR.
IS THERE A DELAY IN REBOUNDS AND
CRISPR MAKE AN IMPACT ON THE
CRISPR RESERVOIR.
WE GAVE THE INFUSION AND ONE
WEEK LATER WE REMOVED ART AND
THEN WE MONITORED THE ANIMALS
EVERY THREE DAYS OR RETURN OF
VIRAL LOAD AND DID THIS WITH THE
GROUP AT NCI FREDERICK.
SO THANKS FOR THAT.
ONCE THE VIRAL LOAD RETURNED WE
DID AN EXTENSIVE NECROPSY.
THE SLIDE IS BUSY BUT SHOWS THE
PROCESS ONCE WE HAVE THE ANIMALS
BECAUSE OTHER LABS ARE DOING
DOWN STREAM ASSAYS.
MY LAB AT RUTGERS IS IN CHARGE
OF THE PROCESSING AND WE RUN THE
STUDIES ANALYZING THE VIRAL
LOADS CHEMISTRIES AND BIO
DISTRIBUTION AND THIS SHOWS THE
INTERACTION WITH THE LARGER
GROUP FOR DOWN STREAMS INCLUDING
CASE WESTERN AND UCLA FOR THE
BAR CODES AND TEMPLE FOR THE
EXCISION.
WE LOOKED AT THE CHEMISTRIES AND
SEE DIFFERENCE AFTER CRISPR FOR
THE GROUPS.
WE ALSO SAW SOMETHING SIMILAR
WITH THE CDC COUNTS AND DIDN'T
SEE SIGNIFICANT CHANGES IN THE
GROUPS.
AND LAST WE LOOKED AT FLOW
ANALYSIS TO LOOK AT CD4, CD 8
AND MONOCYTES AND DIDN'T SEE
SIGNIFICANT DIFFERENCE BETWEEN
THE CONTROL AND CRISPR GROUPS.
OUR LAB PERFORMED CAS 9 DNA AND
RNA FOR DISTRIBUTION AT
NECROPSY.
WE ADD AN EXTENSIVE AMOUNT OF
ORGANS AND TISSUES AND SHOWED IN
BOTH THE CRISPR TREATED AND
CONTROLS WE HAD GOOD
DISTRIBUTION OF THE VECTOR AND
THE CONTROL HAS CAS 9.
AND KAMALSHOWED THE COMMISSION
DATA.
IT'S BY DISTRIBUTION AND SHOWED
IT EXCISES TISSUE AND WHAT WE
ARE INTERESTED IN IS WHAT
HAPPENS ONCE WE REMOVE ART.
YOU'LL SEE FROM THE GRAPH ONCE
WE REMOVED ART VIRAL LOAD CAME
BACK QUICKLY.
IN ALL THE ANIMALS.
THE CRISPR AN CONTROL ANIMALS.
BUT IF WE MADE AN IMPACT ON THE
TIME TO REBOUND THAT WOULD HAVE
BEEN THE HOME RUN.
SO WE DIDN'T GET THE HOME RUN
BUT NOW WE'RE LOOKING AT THE
TISSUE DIVERSITY, VIRAL LOADS
AND RESERVOIRS AND SO WE REALLY
THINK THAT'S WHERE WE'LL GAIN A
LOT OF INFORMATION AND THERE'S
ALWAYS WAYS TO IMPROVE.
THAT'S WHAT WE'RE WORKING ON
NOW.
AND SO ONE OF THE ASSAYS WE'RE
LOOKING AT IS THE IPDA ASSAY.
SO JOHN CARN'S LAB AND BEN
DEVELOPED TWO ASSAYS.
THE TOP ASSAY IS THE IPGA
SLIGHTLY MODIFIED FROM THE LAB
AND THE PRIMER WAS MODIFIED AND
DEVELOPED A FOUR-COLOR DDPCR
ASSAY THAT DETECT THE INTACT PRO
VIRUS AND THE BAR CODED REGION
BETWEEN BPX AND BPR AND THE
CLEAVED REGION.
IT'S BETWEEN PSI AND GAG SO ONCE
IT'S CLEAVED YOU CAN GET
INFORMATION OWN THAT AS WELL.
THIS IS THE ASSAY WE'RE USING.
AND EVERYONE KNOWS WHAT THE IPDA
ASSAY IS BUT JUST TO KNOW IN THE
GREEN THOSE ARE INTACT VIRUSES
AND SIV WE HAVE INTACT PROVIRAL
DNA.
AND DETECTION OF THE FOUR SIV
TARGETS.
THE TOP SHOWS THE TRADITIONAL
IPDA WHICH WE SHOWED THE INTACT
VIRUS.
THE BOTTOM SHOWS WE CAN DETECT
IN THE INTACT VIRUS WE CAN
DETECT THE BAR CODES AND DETECT
THE PSI TO GIVE THE SKWIGS --
EXCISION ANIMALS AND MOST THE
VIRUS IS INTACT AND THE INTACT
VIRUS, 95% FROM THE BAR CODE AND
ALSO 90% OF THE BAR CODED ARE
ALSO INTACT.
SO THIS IS GOING TO GIVE US GOOD
POWER TO LOOK AT THE CODE AND
THE DNA BAR CODE.
THE STUFF WE WERE TALKING ABOUT
BEFORE WAS DNA AND SEQUENCE THE
DNA BAR CODE IN THE STUDIES.
AND THE CONCLUSIONS ARE THAT WE
USED THIS.
IT WAS SAFE.
WE DEVELOPED A NUMBER OF ASSAYS
FOR SIV BUT NOW WE NEED TO GET
INTO THE MEAT OF IT.
SO WE HAVE A LOT OF ONGOING WORK
SO THIS IS A LOT OF PROCESS TO
GET UP TO THIS POINT BUT WE INTO
HE HAD TO MEASURE THE INTACT DNA
AND LOOK AT THE DIVERSITY AND
THESE ARE ALL ONGOING AS WE
SPEAK.
THEN WE ARE ALWAYS LOOKING AT
MESSENGER RAN.
I HAVE TWO OR THREE MORE SLIDES.
THE VISION OF OUR APPROACH.
WE THINK NOW THAT COMBINING
IMMUNE MODULATION WHICH IS WHAT
RF2 IS AND PROVIRAL EXTENSION IS
SOMETHING THAT'S GOING TO
HAPPEN.
WE DON'T THINK IT'S GOING TO
HAPPEN ALONE.
I THINK I'VE HEARD THAT A LOT
THIS WEEK THAT IT'S GOING TO TO
BE SOME COMBINATION.
SO OUR FIRST STUDY TO DO THIS IS
GOING TO USE EXPANDED NK CELLS
ALONE OR IN COMBINATION WITH
CRISPR EDITING AND THEN WE'RE
GOING TO USE THE NOVEL ASSAYS WE
DEVELOPED TO LOOK AT THE IMPACT
OF THE VIRAL RESERVOIR.
SO VERY BRIEFLY THERE'S
ESTABLISHED METHODS TO CULTURE N
ENK CELLS AND THEY ARE GAVE
THREE DIFFERENT INOCULATIONS OF
THESE EXPANDED NK CELLS AND THEN
LOOKED AT HOW IT AFFECTED A FEW
VUREMIA AND SHOWED THERE WAS A
.76 LOG DECREASE IN THE VIREMIA
IN THE NK STUDY WE TOOK THE DATA
AND THOUGHT WE WANT TO DO THIS
BUT WE'LL DO THIS IN ANIMALS
THAT ARE HIGHLY SUPPRESSED
ALREADY IN ART AND COMBINE THIS
WITH CRISPR AND USE AUTOLOGOUS
INFUSION AND COMBINE THAT WITH
THE CRISPR TARGET THERAPY IN THE
SIMILAR MANNER.
WE'RE GOING TO LOOK AT NK CELLS
AND MODIFY THEM.
NK CELLS WHEN YOU INFUSE THEM
INTO A MOUSE AND THIS IS DATA
FROM UNPUBLISHED IN A MOUSE AND
TOOK NK CELLS IN THE MOUSE AND
GOES TO THE LIVER, LUNG AND
SPLEEN.
WE NEED TO GET MORE SO WE ARE
GOING TO RETARGET THE NATURAL
KILLER CELLS AWAY FROM THE LIVER
BY KNOCKING OUT THESE AND WE
HAVE A VERY NICE ANIMAL MODEL
USING THE MISTRG-615 AND SEE
WHAT WORKS AND PROBABLY RECREATE
THAT IN THE MONKEYS.
WITH WANT TO TRY TO IMPROVE
PERSISTENCE AND ENHANCE ACTIVITY
AND DO NOVEL WORK ON KNOCKING
OUT FISH THE ONES WE'VE HEARD
ABOUT ALREADY TODAY.
I'LL END THERE WITH
ACKNOWLEDGEMENTS.
>> NOW I'LL TALK TO EDIT BEING
HIV-1 PROVIRAL.
AND WE HAVE FOUR STEPS.
WE START WITH IN SILICO
SCREENING AND MOVING TO IN VIVO
AND THE HUMANIZED MOUSE AND I'LL
GO THROUGH THIS ROAD MAP.
AND TO FIND THE BEST GUIDE RNA
WE SCREENED AND USED THE NNGRRT
AND LOOKING THE AND INITIAL
SCREENING INVOLVED IN SILICO
LOOKING INTO THE TOOLS WE USE
FOR OFF TARGETING AND DIFFERENT
DATABASES FOR INSTANCE.
WHEN WE DO THESE TYPES OF
SCREENING WE LOOK AT THE BROAD
SPECTRUM AND INITIAL SCREEN IS
IDENTIFIED 14 GUIDE RNAs WITH
HIGH CONSERVATION.
FROM THERE, WE MOVE INTO IN
VITRO CELL LINE SYSTEMS WHERE WE
HAVE MYELOID CELLS LATENTLY
INFECTED VIRUS AND LOOK AT
VARIOUS ASSAYS.
ONE IS THE EDIT ACTIVITY IN
SINGLE GUIDE RNA ACTIVITY OR
INTERFERENCE AND HAVE GUIDE RNAs
WITH ACTIVITY.
THEN WE MOVED FOR FURTHER
VALIDATION.
THESE ARE THE BEST SO FAR
IDENTIFIED FOR GUIDE RNAs OVER
THE 500 GUIDE RNAs WE SCREENED.
WE HAVE COLLABORATION BETWEEN
TEMPLE AND DREXEL UNIVERSITY.
THE BLOOD IS COLLECTED FROM THE
DONORS AT DREXEL AND PBMCs OR
FLESH BLOOD FOR CD4 ISOLATION
AND AT TEMPLE WE FIRST SCREEN
FOR ON TARGET ADDITIVE WHICH IS
INVOLVING SINGLE GUIDE RNA
ANALYSIS AND DUAL GUIDE RNA
COMBINATION IN VARIOUS
COMBINATIONS AND DOING THE
EXCISION ASSAY AND ALL THE
SAMPLES WE COLLECT FROM THE
CELLS GETS TO THE DREXEL FOR
FURTHER ANALYSIS FOR SEQUENCE
AND MOVING INTO THE
COMPUTATIONAL ANALYSIS FOR
QUANTITATIVE ANALYSIS.
THIS IS AN EXAMPLE OF THE ICE
ANALYSIS AND WE HAVE VARIOUS
DONORS AND AMPLIFY THE REGION WE
TARGET AND YOU END UP WITH A
SCORE FROM BETWEEN 0% AND 98%
ROUGHLY.
THAT'S THE RANGE DETECTED FROM
VARIOUS OWNERS.
AND THIS IS AN EXAMPLE OF THE
COMMISSION ASSAY WHERE YOU
COMBINE THEM IN VARIOUS
COMBINATIONS AND YOU SEE
EXAMPLES HERE FOR PSI GAG AND
THE OTHERS AND REPRESENTS THE
TRUNCATED REGION BETWEEN THE
SITES AND YOU VERIFY FOR THE
EXCISION WE ALSO MOVED TO THE
FUNCTIONAL ASSAYS WHERE IN OUR
INITIAL COHORT WE HAD CONTROL
SAMPLES AND ASSESSED FOR SEVEN
DAYS BUT A COUPLE DONORS WE
TESTED SHOWED REBOUNDS FROM THE
BEGINNING AND WE WERE ABLE TO
CHECK THE ATI ASSAY WHERE WE HAD
THE VIRAL REBOUNDS AND WE PUT
THE CELLS ON THE ART AND CHECKED
THE VIRAL LOAD.
INTERESTINGLY ALL THE
COMBINATION SHOWED STRONG
SUPPRESSION IN THE EX VIVO
SETTINGS FOR HIV AND IN
COLLABORATION WITH DREXEL GROUP
AND CAN DETERMINE ON AND OFF
TARGET ASSESSMENT OF THE EACH
INDIVIDUAL GUIDE RNA.
IN SUMMARY FOR EX VIVO WE
PERFORMED STUDIES ON SINGLE
GUIDE ANALYSIS TO EXCISION
ASSAYS AND COMPATIBILITY ASSAY
AND QUANTITATIVE ASSAYS AND
TARGET ANALYSIS AND I DON'T HAVE
TIME TO GO OVER ALL THE RESULTS
BUT SO FAR WE COMPLETED
SCREENING FOR THE FOUR CANDIDATE
GUIDE RNAs IN CD4 T CELLS FROM
16 DONORS AND FINALLY CAME TO A
CONCLUSION OUR COMBINATION
WORKED BEST AMONG THE OTHER
COMBINATIONS SO FAR.
THESE ARE THE GUIDE RNAs.
I'D LIKE TO HIGHLIGHT QUICKLY WE
INITIATED STUDIES INTO THE
CRISPR GENE EDITING AND DREXEL
AND CASE WESTERN RESERVE.
WE LOOK IN OUR INITIAL STUDIES
WE LOOKED INTO THE INHIBITORS
AND USED DIFFERENT ONES IN CD4
PLUS T MEMORY CELLS.
THESE ARE PRELIMINARY AND WE'RE
EXPANDING OUR DONORS AND SAMPLES
BUT INTERESTINGLY THE TAKE HOME
MESSAGE IS WITH THE EPZ WE SEE
LIKE INCREASED ADDED ACTIVITY
COMPARED TO THE UNTREATED CELLS
THOUGH WE SAW VERY HIGH EXCISION
ACTIVITY WITH THE UNC.
WE ARE STILL ANALYZING THE DATA
AND IT'S QUITE EXCITING THAT
EPIGENETICS IN THE CELLS WHERE
VIRUS ARE LATENT AND EPIGENETIC
CONTROL IS IMPORTANT.
SO IN SUMMARY WE STUDIED IN
SILICO AND COMPLETED EX VIVO
STUDIES AND NOW CURRENTLY WE ARE
WORKING ON THE VECTOR SYSTEMS IN
ADDITION TO AAV.
WE'RE WORKING ON A NEW LENTI
VIRAL VIRUS WITH SPECIFIC
DELIVERY IN OUR HUMANIZED MOUSE
MODEL IN THE UPCOMING YEAR.
WITH THIS I WOULD LIKE TO STOP.
THANK YOU.
GOOD AFTERNOON I'M ZACHARY P
PIE -- PIERRE AND I WON'T COVER
THE OTHER STUDIES BEYOND THANK
YOU FOR INVITING US.
WE APPRECIATE IT.
WHAT ARE HIGHLIGHTS?
AS THE PROGRAM MANAGER I MEET
REGULARLY WITH THE COMMUNITY
ENGAGEMENT TEAM USUALLY
BIWEEKLY.
MY BACKGROUND IS IN SOCIAL
SCIENCES I'M MORE OF A PUBLIC
HEALTH PERSON NOT A BENCH
SCIENTIST SO IT'S A NICE
PIPELINE BETWEEN OUR COMMUNITY
ENGAGEMENT EFFORTS AND THE
MANAGEMENT OF THE GROUP.
WE CONTINUE TO HAVE REGULAR
MEETINGS.
ANYONE EXPERIENCED IN RUNNING
CABS KNOWS JUST HOLDING THE
MEETING IS HALF THE BATTLE AND
HAVE CONSISTENT CAB MEMBERS
SHOWING UP AND GIVING INPUT AND
THOUGHTS AND I APPRECIATE THAT.
ONE THING WE DID THIS YEAR WHICH
WAS INTERESTING AND GAVE US
VALUABLE INFORMATION WE SET TIME
ASIDE FOR OUR CAB TO MEET
PRIVATELY WITH THE SAB ADVISORS
AND NIH ADVISORS AND IT SEEMS
LIKE THAT WAS A FRUITFUL
CONVERSATION.
WE LOOK FORWARD TO DOING THAT
AGAIN IN THE FUTURE.
AND SO A MAJOR PROJECT WE'VE
BEEN WORKING ON IS THE CREATION
OF THE COFFEE WITH CRISPR FOR
CURE EDUCATION VIDEO.
THIS AGED ME AND THIS WAS A
PROCESS.
WE MET WEEKLY FOR A LONG TIME TO
DISCUSS WHAT WERE THE NEEDS OF
THE VIDEO AND WHAT THEY NEED TO
LOOK LIKE AND REFLECT WHAT ARE
YOU CONCERNED ABOUT HIV AND CURE
RESEARCH AND KNOWLEDGE ABOUT
CRISPR AND WHAT ARE YOU HOPING
TO SEE FROM OUR RESEARCH.
WE FEATURED THIS AS THE MAY
COMMUNITY MEETING AND GOT GREAT
FEEDBACK AND IF YOU WERE ON THE
CALL AND GAVE YOUR THOUGHTS
THANK YOU SO MUCH AND WE
APPRECIATE IT AND HOPE YOU SEE
YOUR FEEDBACK INTEGRATED IN THE
VIDEO.
IF YOU WANT TO SEE THE VIDEO AND
I RECOMMEND IT WE SPENT A LOT OF
TIME ON THIS IT'S CRISPR FOR
CURE.org AND YOU CAN WATCH THIS
VIDEO.
HERE'S A SCREEN SHOT OF WHAT IT
LOOKS LIKE.
WE HAVE RESEARCHERS ON THE LEFT
AND CAB MEMBERS ON THE RIGHT AND
IF YOU'RE INTERESTED IN BEING
INVOLVED REACH OUT TO OUR
COMMUNITY ENGAGEMENT
COORDINATOR.
AGAIN, PLEASE WATCH THE VIDEO
AND REACH OUT TO KATE IF YOU'RE
INTERESTED.
WE'VE DONE A NUMBER OF
EDUCATIONAL OUTREACH IN THE PAST
YEAR.
OUR FAVORITE WAS PROBABLY A
JOINT EVENT WITH BEAT HIV.
IN TIME FOR PRIDE MONTH IN JUNE
WE TOOK SPACE IN THE WILLIAM
WHITE COMMUNITY CENTER AND
SCREENED A SHORT DOCUMENTARY BY
A LOCAL HIV ACTIVIST AND HAD A
SHORT DISCUSSION WITH HER AND
HER EXPERIENCE AND THEN INVITED
RESEARCHERS FROM CRISPR AND BEAT
TO ENGAGE IN A SPEED DATING TYPE
OF EVENT.
YOU SEE DR. ALBERTO AND
COMMUNITY MEMBERS AT DIFFERENT
TABLES AND I THINK WE WERE GIVEN
FIVE MINUTES AND AT THE END
SCIENTISTS ROTATED AROUND AND
ENGAGED IN ORGANIC CONVERSATIONS
WITH COMMUNITY MEMBERS ON WHAT
YOU'RE WORK BEING ON AND WHAT
ARE YOU HOPING FOR FROM THE
FIELD AND WHAT HAVE YOU SEEN
PREVIOUSLY YOU DON'T THINK IS
WORTH INVESTIGATING.
IT WAS AN ORGANIC CONVERSATION.
COMMUNITY MEMBERS HAD GOOD
THINGS AND RESEARCHERS HAD GREAT
THINGS TO SAY AND NOT JUST
BECAUSE WE PROVIDED A LOT OF
FOOD.
AND AT THE END WE HAD A
COLLECTION OF NICE ART ON LOAN
TO US AND THIS ADDRESS CREATED
BY STUDENTS AND ARTWORK FOR
COLORS AND MY CO-CHAIR AND WE
WERE INVITED TO PRESENT AS A
PANEL DISCUSSION ON HIV CURE
RESEARCH AT THE PHILADELPHIA
PREVENTION SUMMIT.
AND DIRECTOR OF COLORS HOSTED A
COMMUNITY EVENT WITH A PATIENT
AND IT WAS A COMMUNITY
CONVERSATION BETWEEN ADAM AND
OTHERS ABOUT HIV AND WHAT HIS
EXPERIENCE HAS BEEN.
WE WERE GRACIOUSLY INVITED TO
PARTICIPATE AND I WAS ABLE TO GO
AND MEET WITH COMMUNITY MEMBERS
AND HEAR THEIR FEEDBACK AND TALK
ABOUT OUR RESEARCH AND IT WAS A
VALUABLE EXPERIENCE AND THANK
YOU FOR INCLUDING US.
THE WORLD OF CRISPR IS MOVING AT
A BREAK NECK SPEED AND THERE'S A
LOT OF INFORMATION AND WRONG
INFORMATION OUT THERE.
WE WERE SPEARHEADED AT DREXEL
UNIVERSITY AND WE PUT TOGETHER A
SURVEY ASSESSING KNOWLEDGE,
ATTITUDES AND BELIEFS IN HIV
CURE AND CRISPR.
THE GOAL IS TO ROLE THIS OUT AT
STAFF MEETINGS OF OUR PARTNER
ORGANIZATIONS AND SIMILAR
ORGANIZATIONS THAT SERVE
INDIVIDUALS LIVING WITH HIV.
AND WE KNOW WHERE PEOPLE RECEIVE
CARE IMPACTS THEIR ATTITUDES AND
WE ALSO SAW EARLIER THAT
PROVIDERS HAVE A RANGE OF
RESEARCH AND SO THE HOPE IS WE
CAN GET GOOD INPUT HOW OUR
MEDICAL PROVIDERS ARE FEELING
ABOUT HIV, HIV CURE AND CRISPR.
WE WANT TO INCLUDE ALL LEVELS
BECAUSE IT'S NOT JUST DOCTORS OR
NURSE PRACTITIONERS BUT YOUR
PHLEBOTOMISTS AND THE PEER
NAVIGATOR AND ALL THESE
INDIVIDUALS IN THE CARE TEAM
IMPACT HOW PEOPLE RECEIVE HEALTH
INFORMATION AND IMPACT WHAT
THEY'RE ABLE TO OR WILLING TO
ENGAGE WITH.
OF COURSE IF YOU'RE INVOLVED IN
CABS RECRUITMENT IS AN ONGOING
CONCERN AND HOPING TO ROLL THIS
OUT AND USE IT AS A RECRUITMENT
TOOL TO RECRUIT NEW CAB MEMBERS.
THAT'S THE END OF MY
PRESENTATION.
THANK YOU.
>> I KNOW WE'RE KEEPING YOU FROM
YOUR DINNER BUT IF ANYONE HAS A
QUESTION FOR THE CRISPR FOR CURE
PLEASE COME UP TO THE
MICROPHONE.
>> HELLO.
SO IF I UNDERSTOOD YOUR MACAQUE
DATA YOU HAD 10 ANIMALS ON ART
FOR TWO YEARS AND THE CRISPR
TREATMENT I THINK YOU USED 10 TO
THE 13 PER KIG A TYPICAL DOSE IN
SYSTEMIC AAV GENE THERAPIES AND
USED ATI AND SAW NO EFFECT AND
THAT MATCHES FROM THE DATA FROM
THE IN HUMAN TRIALS I BELIEVE
THREE PATIENTS UNDERWENT ATI AND
ALL REBOUNDED.
>> THAT'S TRUE BUT ONE HAD A
DELAYED REBOUND.
I'M NOT SURE THE EXACT DATA.
SIX MONTHS, 16 WEEKS.
>> I'M WONDERING WHAT THE
COMPELLING EVIDENCE IS FROM YOUR
MACAQUE STUDIES.
IT MAKES YOU WANT TO GO INTO
HUMANS.
I'M NOT SEEING IT.
>> LIKE I SAID IN THE TALK WE
HAVE A LOT OF WORK TO DO.
I THINK --
>> WITH RESPECT YOU WENT INTO
HUMANS PRESUMABLY WITH A LOT OF
WORK SO WONDERING WHAT THE
COMPELLING EVIDENCE WAS AND NOT
YOU PERSONALLY BUT THE RELATED
STUDIES THAT WENT INTO HUMANS.
>> I DON'T KNOW IF KAMAL WANTS
TO COMMENT.
>> MAYBE I CAN ASK IT A
DIFFERENT WAY SLIGHTLY LESS
AGGRESSIVE, MAYBE I CAN
APOLOGIZE.
MAYBE YOU CAN EXPLAIN TO ME WHAT
YOU WERE HOPING TO SEE THAT
WOULD BE CONVINCING ENOUGH FOR
YOU THAT IT SHOULD HAVE ANOTHER
ROUND IN HUMANS?
>> I CAN ANSWER THAT ONE.
>> WE WOULD LIKE TO SEE WHEN YOU
STOP ART THE VIRUS DOESN'T COME
BACK.
>> WELL, YOU'VE DONE THAT
EXPERIMENT TWICE IN HUMANS.
>> WHAT I SAID IN MY TALK AND
HOPEFULLY I'M SORRY I RUSHED
THROUGH A LITTLE BIT WE'RE
HOPING TO SEE A DECREASE IN THE
VIRAL DIVERSITY IN THE TISSUE
RESERVOIR RATE BECAUSE IF YOU
HAVE IN THE CIRCULATION IF YOU
HAVE ONE VIRUS THAT GETS OUT
THERE AND REBOUND'S GOING TO GO
UP.
DID WE MAKE AN IMPACT ON THE
TISSUE RESERVOIR, RIGHT.
THAT'S ONE THING WE HAVEN'T
EXAMINED FULLY TO SEE IF THAT IS
IF WE HAVE DONE THAT.
THAT'S WHAT I SAID.
I THINK VIRAL REBOUND AND
ESPECIALLY IN AN SAB 239 MACAQUE
MODEL TO HAVE VIRAL REBOUND AS
YOUR GOAL IS LIKE TO STOP VIRAL
REBOUND IS LIKE UP HERE.
SO I THINK WE HAVE TO MAKE THOSE
STEPS THAT SHOW DOES IT AFFECT
DIVERSITY OF THE RESERVOIR
BEFORE WE CAN GET UP THERE.
AND THEN HONESTLY WE USE AAV9
AND EVERYONE IN THE FIELD KNOWS
THAT'S NOT PROBABLY THE BEST
DELIVERY BUT IT'S THE BEST
DELIVERY WE HAVE AT THE MOMENT.
SO IT'S A GOOD START.
I THINK BETTER DELIVERY.
IF WE DO SEE THE VIRAL LOAD IN
TISSUES IS DECREASED A COUPLE
LOGS TO ME THAT WOULD BE
INTERESTING.
>> OKAY, GOOD LUCK.
>> ARE THERE ANY OTHER
QUESTIONS?
EITHER IN THE ROOM OR ONLINE?
OKAY.
>> OVER ALL THE PROBLEM OF
EXERTION IS A DIFFICULT ONE BUT
MY QUESTION WAS MORE WHAT ARE
YOU DOING TO ADDRESS THAT?
AT WHAT STEP IS THE DISCONNECT
BETWEEN YOUR TREATMENT AND THE
ATI?
I THINK YOU CAN PROBABLY LEARN A
LOT FROM THESE STUDY.
I WOULDN'T EXPECT LIKE A 100%
CURE.
IT'S A DIFFICULT PROCESS.
I WOULD LIKE TO SEE THE STEPS IN
BETWEEN ADDRESSED AND WHAT COULD
BE IMPROVED.
>> THAT WAS THE FIRST TIME THAT
WE DID THE EXPERIMENT AND
THERE'S A LOT OF COLLABORATION
THAT NEED TO BE DONE.
ONE IS THE DELIVERY SYSTEM AND
THOSE OF THE DELIVERY SYSTEM
WE'RE USING AND MORE THE DESIGN
OF THE EXPERIMENTS.
FOR EXAMPLE HERE WE DIDN'T
PROVIDE ENOUGH TIME FOR CRISPR
TO CIRCULATE IN THE BODY.
ONLY SEVEN DAYS.
MAYBE YOU SHOULD DO IT LONGER.
IT'S A START OF THE JOURNEY THAT
I THINK IS GOING TO TAKE US TO
THE PLACE THAT WE WOULD LIKE TO
BE.
BUT IT'S A FIRST TIME
EXPERIMENT.
FIRST TIME YOU'RE SHOWING IN THE
LYMPH NODE YOU CAN EXCISE PART
OF DNA BY A LOUSY DELIVERY
SYSTEM SUCH AS AAV9 AND IF YOU
IMPROVE IT INCREASES THE
EFFICIENCY OF THE EXCISION OR
EDITING.
EVEN IN THE BRAIN WE SHOWED THE
VIRAL DNA IN PLACE AND WE HAVE
TO WAIT A LITTLE BIT ON THE
OTHER DATA COMES AND FROM
INFECTED ANIMALS.
>> WE WANTED THE CRISPR TO BE
THERE WHEN WE CATCH THE VIRUS
REBOUND AND THIS SIV IS AN
EXTREMELY AGGRESSIVE VIRUS.
>> THERE'S A SINGLE APPROACH
THAT'S CHALLENGING.
WE SHOW PLENTY OF DATA AND
CRISPR SYSTEM CAN EXCISE AND
DEACTIVATE HIV BUT DELIVERY IS
ONE KEY.
THAT'S WHY WE'RE INTEGRATING
INTO THE SYSTEM WITH CD4
SPECIFIC DELIVERY.
AND ALSO WE WORK ON AN VECTOR
WHICH IS GOING TO ONLY GO TO CD4
CELLS.
I THINK DELIVERY VECTOR
IMPROVEMENT WILL CHANGE THE
OUTCOME.
AND ALSO WE ARE WORKING ON
STRATEGIES TO COMBINE IN
MULTIPLE APPROACHES LIKE
TARGETING NK CELLS OR NK
MEDIATED THERAPY COMBINED WITH
CRISPR AND MAYBE THE CELLS WILL
TAKE THAT OUT AND THOSE ARE
APPROACHES WE BELIEVE WILL
IMPROVE SIGNIFICANTLY.
>> YOU WILL USE THE ANIMALS THAT
REBOUNDED AND PUT CRISPR ON THEM
AGAIN.
TREAT THEM OVER AND OVER.
>> WE CANNOT DO IT WITH THE
CURRENT DELIVERY SYSTEM BECAUSE
BUT WITH THE NEW DELIVERY SYSTEM
WE'RE DEVELOPING WE MIGHT BE
ABLE TO DO THAT.
>> SORRY, WE'RE A LITTLE BIT OUT
OF TIME AND I SEE SHARON YOU GET
THE LAST QUESTION AND THEN THE
BREAK.
>> ACTUALLY I WAS GOING TO ASK
ABOUT THE ROLE OF REPEATED
DOSING INFECTION.
DO YOU WORK WITH DELIVERY IN
mRNA OPPOSED TO THE ARB SYSTEM.
>> AND WE'RE CONTAINING THE
COMBINATION AS WELL AS WE CAN
PACKAGE IN OTHER PARTICLES AND
WHERE HE ARE WORKING ON THE
MODELS AND GREAT POSTERS OUT
THERE FROM OTHER GROUPS
DEVELOPING SIMILAR SYSTEMS AND
WE'RE ADOPTING THOSE.
>> THANK YOU AND CONTINUE THE
CONVERSATION OVER DINNER.
SO LET'S THANK CRISPR FOR CURE.
THANK YOU.
>> I HAVE AN ACKNOWLEDGEMENT
THERE'S A PROGRAM THIS EVENING
IF YOU'RE INTERESTED UP HERE ON
THE SCREEN.
IT'S AN ORGANIZED COLLABORATION
BETWEEN THE CENTER FOR AIDS
RESEARCH AND OTHERS.
PLEASE FEEL FREE TO GO TO THIS
EVENT AND FOR TOMORROW WE START
THE SESSION AT 8:30 EXACTLY.