>> GOOD AFTERNOON AND WELCOME TO
THE WEDNESDAY AFTERNOON LECTURE
SERIES, I'M MICHELLE ANTWON, THE
ACTING CHIEF ON NIAAA TRIPLE
NEURON CIRCUITS AND IT'S MY
HONOR TO INTRODUCE DR. FENG,
DR. FENG HAS CONTRIBUTED TO HOW
NEUROABNORMALITIES CONTRIBUTE TO
NEURODEVELOPMENTAL DISORDERS,
SUCH AS AUTISM, RHETT'S
SYNDROME, PSYCHIATRIC DISORDERS
LIKE SKITS SKITS.
FOR THOSE REMOTING WATCHING WE
ENCOURAGE TO YOU PARTICIPATE IN
THE Q&A SESSION OF TODAY'S
PRESENTATION.
YOU CAN DO SO BY CLICKING ON THE
BUTTON JUST BELOW YOUR VIDEOCAST
WINDOW THAT SAYS SEND LIVE
FEEDBACK.
CLICK ON THAT TYPE IN YOUR NAME
AND QUESTION AND WE WILL RELAY
THIS TO DR. FENG AT THE END OF
HIS LECTURE.
YOU CAN SUBMIT A QUESTION AT ANY
TIME AND I HOPE YOU DO SO.
SO FOR THOSE OF YOU IN
LIPSETT AMPHITHEATRE TODAY, YOU
CAN USE THE MICs ON THE LEFT
AND RIGHT OF THE ROOM TO ASK
QUESTIONS.
ALSO, WE ARE OFFERING CONTINUING
MEDICAL EDUCATION CREDITS FOR
THIS PRESENTATION.
SO THE CME CODE FOR TODAY IS
57683.
I WILL REPEAT THAT IT'S 57683.
OUR OUR SPEAKER TODAY IS
DR. GUOPING FENG, ASSOCIATE
DIRECTOR OF THE Mc GOVERN
INSTITUTE AT THE MASSACHUSETTS
INSTITUTE OF TECHNOLOGY ALSO THE
JAMES W. AND PATRICIA PROFESSOR
IN BRAIN AND COGNITIVE SCIENCES,
HE IS THE DIRECTOR OF THE HOP E.
TAN AND YANG K. LEE CENTER FOR
AUTISM RESEARCH, HE'S A DIRECTOR
OF MODEL SYSTEMS AND
NEUROBIOLOGY AT THE STEME CENTER
FOR RESEARCH AT THE BROAD
INSTITUTE.
DR. FENG, RECEIVED HIS Ph D AT
BUFFALO UNDER THE THESIS MENTOR
OF OF LINDA HALL, WHERE HE
WORKED ON GPC R50 AND
DROSOPHILA.
HE DID A POST DOC WITH JOSH
RACINs AT WASHINGTON SCHOOL OF
MEDICINE WHERE HE CONTINUED TO
HONE AND DEEPEN HIS KNOWLEDGE
ONIN SYNAPTIC FUNCTION ON
INVERTEBRATES.
IN 2001, HE MOVED TO DUKE
UNIVERSITY WHERE HE BECAME AN
ASSIST APT PROCESSOR IN THE
DEPARTMENT OF NEUROBI
ALGORITHMS.
WHILE THERE HE ESTABLISHED A
POWERFUL BLUEPRINT FOR HOW WE AS
NEUROSCIENTISTS CAN USE GENETIC
MODELS TO REPLICATE COMPLEX
HUMAN DISEASES LIKE OBSESSIVE
COMPULSIVE DISORDER AND HE
DEMONSTRATED THAT TREATMENTS
THAT ARE EFFECTIVE IN HUMANS CAN
BE SIMILARLY APPLIED TO REVERSE
THESE SYMPTOMS MODELED IN MICE.
ALTHOUGH HIS WORK ILLUSTRATED
HOW TO ESTABLISH THE CRITICAL
STEPS IN MODELING DISEASE, HE
ESTABLISHED OUR ABILITY TO
PRODUCE FEASIBILITY,
UNSTRUCTURALLIABILITY AND
UNPREDICTABILITY IN MICE.
HIS FRAMEWORK LAY THE FOUNDATION
OF COURSE FOR THE NEXT ESSENTIAL
STEP WHICH IS CONNECTING THE
SPECIFIC NORMAL BEHAVIORIAL
SYMPTOMS TO THE UNDERLYING GENS
AND SYNAPTIC MECHANISMS AND THAT
GROUND BREAKING WORK WAS
PUBLISHED IN NATURE IN 2007 SO
AS A YOUNG GRADUATE STUDENT AT
THE TIME I WAS DEEPLY INSPIRED
BY DR. FENG'S WORK AND I
CONTINUE TO DO SO TODAY AND IN
FACT MANY OF US IN THIS ROOM
CONTINUE TO APPLY DR. FENG'S
STEAP BY STEP STREAMLINE
APPROACH TO TALK ON SOME OF THE
MOST DIFFICULT AND COMPLEX
BIOLOGICAL QUESTIONS IN OUR OWN
WORK TOWARDS UNDERSTANDING DEC
MECHANISM AND IDENTIFYING NEW
TREATMENTS.
IN 2010 DR. FENG JOINED THE
Mc GOVERN INSTITUTE WHERE HE
REMAINS TODAY.
AND SINCE THEN HE FULLY REFINED
HIS ALREADY RIGOROUS FRAMEWORK
TO EXPLORE THE RULE OF MUTATIONS
IN THIS SHENG 3 GENE AND LINKING
BEHAVIORS LIKE AUTISM IN MOUSE
MODELS, HIS RESEARCH ALSO HELPED
AFFIRM A PROMISE OF GENE THERAPY
AS A TREATMENT STRATEGY FOR
NEURODEVELOPMENTAL DISORDERS.
DR. FENG HAS CONTINUED TO PUSH
THE BOUNDARIES OF NEUROSCIENCE
BY EXTENDING HIS RESEARCH
FRAMEWORK TO NONHUMAN PRIMATE
MODELS AND EXCITING AND
AMBITIOUS THAT YOU HEAR MORE
ABOUT IN HIS TALK.
HIS WORK CONTINUES TO SHAPE HOW
WE UNDERSTAND AND TREAT COMPLEX
HUMAN DISEASES.
DR. FENG HAS RECEIVED NUMEROUS
HIGH PROFILE AWARDS ALONG THE
WAY INCLUDING THE Mc KNIGHT
TECHNOLOGICAL INNOVATIONS IN
NEUROSCIENCE AWARD, THE HEART
WELL INDIVIDUAL BIOMEDICAL
RESEARCH AWARD, HE WAS AFTER THE
ACCIDENT LECTED TO THE NATIONAL
ACADEMY OF MEDICINE IN 2003, THE
AMERICAN ACADEMY OF ARTS AND
SCIENCES IN 2019 AND THE
NATIONAL ACADEMY OF SCIENCES IN
2013, IN 2023.
THE TITLE OF DR. FENG'S TALK
TODAY IS DISSECTING
NEUROBIOLOGICAL MECHANISMS OF
ASD FROM GENES TO CIRCUITS.
PLEASE HELP ME WELCOME
DR. GOUPING FENG.
[ APPLAUSE ]
>> THANK YOU MICHELLE FOR A VERY
GENEROUS INTRODUCTION, IT'S A
REAL HONOR TO BE HERE AND TO BE
IN THIS POWER HOUSE OF RESEARCH
AND SINCE I WAS A GRADUATE
STUDENT HERE IN U.S., I ALWAYS
ADMIRED THE UNBELIEVABLE WORK
DONE HERE AND ACTUALLY AT THE
END OF MY TALK, YOU WILL SEE I
STILL FOLLOW SOME OF THE EARLY
DISCOVERIES AND FROM NIH CAMPUS
BECAUSE I'M ALWAYS VERY GRATEFUL
FOR THE NIH SUPPORTING ALL THE
EXTRAMURAL PROGRAMS THAT WE RELY
ON THIS.
I THINK WE ARE MORE AND MORE
NEED THIS AT THIS TIME TO PUSH
OUR RESEARCH INTO BENEFIT OF
PATIENT.
AND TODAY I WILL TALK ABOUT SOME
OTHER RESEARCH AND THINKING HOW
CAN WE USE AND BUILD OUR
RESEARCH IN THE PAST, MANY, MANY
DECADES AND EVENT CRULLLY CAN
POTENTIALLY HELP -- EVENTUALLY
CAN HELP PATIENTS RESOLVE NIH
RESEARCH AND MANY OF OUR
SCIENTISTS GOALS AND ALSO OUR
NEUROSCIENTISTS GOAL.
SO THIS IS MY DISCLOSURE ALREADY
SHOWED BY MICHELLE'S SLIDES SO,
IN THE PAST YOU KNOW 20 YEARS,
MORE OF THE BIGGEST VANCES IN
YOU KNOW BRAIN SCIENCE, BRAIN
DISSEREDDER RESEARCH IS THE
IDENTIFICATION OF RISK GENES
BECAUSE AS A SCIENTIST FOR BRAIN
RESEARCH UNLIKE CANCER WHICH IS
ALSO VERY DIFFICULT PROBLEM,
IT'S VERY HARD FOR US TO GET
LIVE BRAIN TISSUES, THE WHOLE
BRAIN TO STUDY, SO WE HEAVILY
RELY ON ANIMAL MODELS, RIGHT, SO
BE ABLE TO USE ANIMAL MODELS, WE
NEED, YOU KNOW GENETIC MODELS
AND GENETIC INFORMATION IS 1 OF
THE KEY THINGS WE CAN ACTUALLY
ENGINEER AND PUT A HUMAN
MUTATION INTO GENETIC MODELS.
USING AUTISM RISK GENE AS AN
EXAMPLE, WE ACTUALLY HAVE -- LET
ME SEE, I CAN -- SO IN THE PAST
2 DECADES WE HAVE IDENTIFIED
MANY, MANY GROUPS, NOT IN MY
LAB, BUT MANY OTHER GROUPS AND
IDENTIFIED HUNDREDS OF RISK
GENES AS SOME ARE MONOGENIC
CAUSES OF ASD GENES AND WITH
THIS GENETIC INFORMATION, WE CAN
GENERATE WITH NEW TECHNOLOGY
ESPECIALLY GENOME EDITING
TECHNOLOGY, WE CAN GENERATE MANY
KIND OF GENETIC MODELS INCLUDING
CELL MODELS, RODENT MODELS AND
THE PRIMATE MODELS, AND THEN USE
THIS NEW TECHNOLOGIES LIKE
CRISPR, AND YOU KNOW SEX
MANIPULATION AND ESPECIALLY
SINGLE CELL TRANSCRIPTOMIC
ANALYSIS, WE CAN HELP US
UNDERSTAND THE NEUROBIOLOGICAL
MECHANISMS MPLET THE HOPE IS
WITH THIS KIND OF STUDY WE CAN
EVENTUALLY EXPLORE THERAPEUTIC
STRATEGIES.
AND SO FOR ASD FOR EXAMPLE, MANY
OF THE GENES ARE ACTUALLY RISK
GENES AFFECT THE SYNAPSE, PLAY
AN IMPORTANT ROLE IN SYNAPSE
ENVIRONMENTAL FUNCTION AND
PLASTICITY.
THIS IS A SLIDE I BORROWED FROM
THOMAS' REVIEW, AND YOU CAN SEE
FROM THESE EXCITED SYNAPSE,
INHIBITOR SYNAPSE, BROWN ARE THE
PROTEINS AND IMPORTANT FOR
SYNAPTIC FUNCTIONING AND
ENVIRONMENT BUT ALSO RISK GENES
FOR AUTISM.
AND SO MY LAB LIKE MANY LABS ARE
VERY INTERESTED IN THE
ENVIRONMENT, FUNCTION AND
DYSFUNCTION OF THE EXCITED
SYNAPSE ESPECIALLY IN THE
POSTSYNAPTIC SITE, SO IN THE
PAST 3 DECADES HAVE CONTRIBUTED
TO THE UNDERSTANDING OF OUR
MOLECULAR ARCHITECTURE OF
POSTSYNAPTIC DENSITY INCLUDING
MARY KENNEDY, MORGON SHENG, AND
DAVID BREDT, AND MANY OTHER
LABS, I GIST LIST A FEW OF THEM.
SO IN THIS VERY SMALL STRUCTURE,
POSTSYNAPTIC DENSITY, WHICH IS
ABOUT 2 MICRONS AND THE
THICKNESS IS 2-NANOMETERS WE
ACTUALLY IDENTIFIED 1500 TYPE OF
PROTEINS AND THAT'S THE IMPACT,
THAT'S WHY I CALL IT
POSTSYNAPTIC DENSITY, CAN YOU
SEE IT HERE WITHOUT ANY STAINING
AND HOW THESE -- SUCH A
MACROMOLECULAR STRUCTURE IS
ASSEMBLED AND DYNAMICALLY
REGULATED IS ALWAYS AN
INTERESTING PHENOMENA TO MANY OF
US AND ESPECIALLY FOR FULL STUDY
SYNAPSES.
SO FOR THIS ASSEMBLY IS CALLED A
SCAFFOLDING PROTEIN, SO THEY ARE
MORE OF THE SCAFFOLD IS A PS95
SHANK, A SCAFFOLDING, EACH OF
THESE PROTEINS HAVE
PROTEIN-PROTEIN DOMAINS AND WITH
THE PS95 INTERACTING WITH THE
PROTEINS WITH THE MEMBRANE
PROTEINS ON THE POSTSYNAPTIC
SIDE INCLUDING THE AMP O AMD
RECEPTORS AND ION CHANEMS AND
MOLECULES, ON THE OTHER HAND YOU
CAN BRING MANY, MANY SIGNALING
MOLECULES TO THE POSTSYNAPTIC
SITE VERY DYNAMICALLY AND OTHER
WITH SHEK, NOT ONLY BRING THE
SIGNALING MOLECULE TO IT BUT
ALSO IT CAN INTERACT WITH YOU
KNOW CYTOSKELETON INTERACTING
PROTEIN.
SO IT PLAY A ROLE IN NOT ONLY
FUNCTION BUT PLASTICITY BOTH IN
THE SIGNALING AND THE FUNCTIONAL
PLASTICITY BUT ALSO STRUCTURAL
PLASTICITY.
AND SO 1 OF THE PROTEINS, WE ARE
REALLY INTERESTED IN MANY OTHERS
AND ALSO IS THE SHENG, FAMILY OF
PROTEINS, THE SHANK IS A ON THE
POSTSYNAPTIC SITE, THE GLUTEA
MITTERGIC SYNAPSES BUT THE SHANK
MORE IMPORTANTLY SHANK 3 AND THE
MUTATION OF SHANK 3 IS
MONOCLONAL, AND CAUSES SEVERE
AUTISM WITH INTELLECTUAL
DISABILITY AND THIS IS THE MAIN
GENE IN THE DERMA SYNDROME WHICH
IS IDENTIFIED BY DR. FEELIN AND
DR. Mc DERM, AND IT IS 22 Q13,
THIS IS VERY SEVERE, THEY HAVE
HYPOTONIA, DEVELOPMENTAL DELAY,
SEVERE INLECTUAL DISABILITY,
MOST IF NOT ALL PATIENT CANNOT
LIVE INDEPENDENTLY AND THEY ALSO
HAVE IMPAIRED LANGUAGE
DEVELOPMENT AND AUTISM SPECTRUM
FEATURES AND THEY ALSO HAVE MANY
OR CO-MORBIDITY ISSUES INCLUDING
SLEEP DISORDERS AND SEIZURE
ANDOR OIJINALLY BECAUSE THIS WAS
MONOGENIC WE COULD MAKE MODELS
AND THEN TRY TO UNDERSTAND WHAT
IS THE MOLECULAR CHANGE THAT YOU
KNOW WE CAN IDENTIFY THEN WE CAN
DEVELOP A THERAPEUTICS, DRUGS TO
TREAT THIS CONDITION BECAUSE
IT'S SO SEVERE.
SO WE -- 1 OF THE FEATURES OF
THIS GENE IS IT'S HIGHLY
EXPRESSED IN THE TRIT UMKC.
THERE ARE 3 SHANKS AND ONLY 3
SHANKS ARE EXPRESSED IN THE
STRIATUM.
THE DYSFUNCTION HAS BEEN
STRONGLY SMLICATED LIAISON --
IMPLICATED IN AUTISM AND OTHER
NAWR O LOGIC DISORDERS, I WILL
GO QUICKLY AND SO WE MADE A
KNOCK OUT OF THE SHANK PROTEIN,
YOU KNOCK OUT SHANK 3 AND YOU
LOSE OTHER SYNAPTIC PROTEINS IN
THE SIDE CLUING AS A SCAFFOLDING
PROTEIN AND THE AMPORECEPTORS
AND ALSO AFFECT FUNCTION AND
REDUCE SYNAPTIC TRANSMISSION AND
MOST OF THE MINUTE FREQUENCY AND
BUT ALSO AMPLITUDE.
THEY ALSO AFFECTED A NUMBER OF
SYNAPSES IN THE -- OUT OF THE
SPINE 1S SO WE FOCUS ON THE
SYNAPSES AND THE SPINE IN THE
MEDIAN AND SPINE NEURONS IN THE
STRIATUM.
SO THIS ANIMAL ALSO HAVE MANY
BEHAVIOR IN HOMOZYGOUS KNOCK
OUT, IN PATIENTS ALL
HETEROZYGOUS, BUT IN MICE THEY
HAVE VERY LITTLE PHENOTYPE, WE
USE HOMOZYGOUS TO STUDY THE
FUNCTION OF SHANK 3, THEY ALSO
HAVE VERY ANIMAL BEHAVIOR,
REPETITIVE, COMPULSIVE, THEY ARE
GROOMING THEIR HAIR OFF AND SKIN
OFF AND THEY ALSO HAVE CHAMBER
ASSAYS, AND THEY DEVELOPED THIS
HERE AT NIMH AND ALSO HAVE
SOCIAL INTERACTION DEFICIT,
SENSORY OVERSENSITIVITY,
ANXIETY, REDUCED LOCOMOTION,
IMPAIR THE MOTOR LEARNING, THIS
IS RELATIVE TO AUTISM DISORDER
ANDS COMORBID CONDITIONS SO WE
CAN IDENTIFY CONVERGING
MECHANISMS THAT ALLOW US TO
DEVELOP A DRUG.
SO WE LOOK AT MOLECULAR AND
CELLULAR DEFECT IN THE ANIMALS,
I'M NOT GOING THROUGH ALL THE
STUDIES, I JUST SUMMARIZE WITH
YOU 1.
WHAT WE FOUND IS FOR EACH
PHENOTYPE, THE THERE IS A
DIFFERENT CELL TYPE INVOLVED AND
DIFFERENT DEFECT INVOLVED, SO
FOR EXAMPLE, THE COMPULSIVE
BEHAVIOR, WE FINALLY REDUCE THE
NEURONAL ACTIVITY INTERACTIVE
PATHWAY, AND SPINE NEURON WHICH
IS THE STRIATUM.
IF YOU CORRECT THE DEFECT IN THE
SPINE, YOU CAN CORRECT THE
REPETITIVE GROOMING BEHAVIOR,
AND FOR SOCIAL INTERACTION, MANY
LAB STUDY AND MY LAB STUDIED
WITH A FORMER POST DOC AND FOUND
THAT THE MAIN STUDY REDUCED THE
EXCITED NEURONS IN THE ACC, IT
IS THE CAUSE AT LEAST 1 OF THE
CAUSE OF THE SOCIAL INTERACTION
DEFICIT.
IF YOU INCREASE ACTIVITY OR
KNOCK OUT SHANK 3 ONLY IN THESE
NEURONS YOU CAN PROVIDE DIRECT
EVIDENCE THAT THIS GROUP OF
NEURONS AT LEAST IS INVOLVED IN
THIS SOCIAL INTERACTION DEFICIT.
WE ALSO SAW SENSORY
OVERSENSITIVITY, IN
COLLABORATION WITH CHRIS AT THE
UNIVERSITY, SHOWED THAT
INHIBITING INTERNEURONS IN THE
SOPHISTICATED MAT O SENSORY
TORTEX IS RESPONSIBLE FOR THE
SENSORY OVERSENSITIVITY.
SO PUT THEM ALTOGETHER, THIS IS
A SINGLE MONOGENIC CAUSE, RIGHT?
EVEN FOR MONOGENIC CAUSE, YOU
HAVE DIFFERENT CIRCUITS,
DIFFERENT CELL TYPES IN
DIFFERENT BRAIN REGIONS FOR
DIFFERENT PHENOTYPES AND IMAGINE
THIS IS POLYGENIC AND WE DON'T
KNOW HOW TO MODEL THEM SO IT
SEEMED LIKE IT'S REALLY
DIFFICULT TO IDENTIFY A
CONVERGING MECHANISM FROM MUCH
OF A MODELS OF AUTISM TO FIND A
THERAPEUTIC TARGET.
SO IT'S NOT SURPRISING BECAUSE
FOR SHANK 3 AND MANY OTHERS IT'S
A TRANSLATIONAL REGULATOR THAT
AFFECTS THE GENES BY BOB
DARNEL'S LAB AT THE UNIVERSITY,
AND IDENTIFY X, AND SHOW AT
LEAST 300 GENES ARE AFFECTED BY
TRANSCRIPTION.
SO FOR SHANK 3 IT'S A SCAF OLDER
PEOPLING PROTEIN INTERACTING
WITH MANY DIFFERENT PROTEINS AND
SO IT'S NOT SURPRISING THAT CAN
YOU IMAGINE IN DIFFERENT CELL
TYPES AND DURING DIFFERENT
ENVIRONMENT TIME IT WOULD
REQUIRE DIFFERENT SIGNALING MECH
NIMS,
NIMS -- MECHANISMS, SO IT WILL
BE DIFFICULT TO FIND CONVERGING
MECHANISMS IN MY VIEW, MY LAB
STUDY, MANY OTHER LABS STUDIED
SHOWED THAT THERE ARE MANY
PATHWAYS HAVE BEEN DISRUPTED, SO
THEN THE QUESTION IS, IF IT'S SO
COMPLEX HOW CAN YOU FIND A CELL
TO TARGET THAT WOULD BENEFIT
MANY AUTISTIC KIDS.
SO HOW CAN YOU TREAT ALL
DIFFERENT SYMPTOMS CAUSED BY
DIFFERENT MECHANISMS?
SO THAT MADE US TO THINK ABOUT
THE CHANGE THE STRATEGY AND WE
THINK THAT MANY OF US INCLUDING
MYSELF THINK THERE MAY BE WE USE
DIFFERENT STRATEGIES, FOR THE
MONOGENIC CAUSE, MAYBE GENE
THERAPY WAS THE NEW TECHNOLOGY
ENVIRONMENT, MAYBE THE BEST WAY
OR AT LEAST 1 OF THE BEST WAYS
TO DIRECTLY CORRECT THE GENETIC
DETHAT ACCOUNT BE POTENTIALLY A
CURE FOR THE MONOGENIC
DISORDERS, ON THE OTHER HAND
MOST AUTISM OR MOST ARE NOT
MONOGENIC, THAT ARE NOT
MONOGENIC, THEY ARE CALLED BY
POLYGENIC FACTORS PLUS OTHER
FACTORS INCLUDING ENVIRONMENTAL,
AND MANY RPGHT FACTORS AND
ENVIRONMENTAL, ALL THIS COULD
CONTRIBUTE TO IT.
FOR THIS WE THINK THAT 1
APPROACH COULD BE BY
UNDERSTANDING NEUROSUBSET OF
CAUSE A MOST PARTICULAR
DEBILITATING SYMPTOM AND BY
CORRECTING THAT WE MIGHT BE ABLE
TO DRAMATICALLY IMPROVE THE
BRAIN FUNCTION, BEHAVIOR AND THE
PATIENT'S LIFE.
SO I WILL USE 2 EXAMPLES, I WILL
USE SHANK 3 AS AN EXAMPLE TO
ILLUSTRATE HOW WE MIGHT BE ABLE
TO MAKE A PROGRESS IN THESE 2
FRONTS.
SO NEURAL DEVELOPMENT DISORDER
SUCH AS AUTISM, THEY ARE MAYBE
ENVIRONMENTAL, MAYBE REVERSIBLE,
SO SOME OF THE EARLY WORK, YOU
KNOW LANDMARK WORK BY ANDRIEA
BURG SHOWED THAT IF YOU KNOCK --
IF YOU PROVIDE THE GENE LATER
ON, IT CAN REVERSE EVERYTHING.
SO I THINK EACH GENE IS
DIFFERENT, DEPENDS WHERE THEY
ARE EXPRESSED AND WHEN THEY ARE
EXPRESSED SO I THINK FOR EACH
GENE WE HAVE TO TEST THEM SO WE
TEST THEM WITH SHANK 3, WE MADE
A MOUSE MODEL THAT USED DOUBLE
FLOX, INCLUDING CODING THE
DOMAIN HAD IS A KEY FUNCTIONAL
DOMAIN IN THE SHAIRCHG 3 PROTEIN
AND SO THESE MICE GREW UP LIKE
KNOCK OUT, THEY HAVE BASICALLY
ALL THE PHENOTYPES I DESCRIBED
BEFORE.
BECAUSE FLOX IS CREE DEPENDENT
YOU CAN USE TAMOXIFEN-CREE CROSS
AND AT WHICH TIME POINT YOU
PROVIDE A FULL SHANK 3 GENE CAN
RESCUE THIS PHENOTYPE.
SO WHAT WE FOUND IS THAT IN
ADDITION, IF YOU TURN UP SHANK 3
GENE BACK ON IN ADULT, YOU CAN
ACTUALLY CORRECT SOME OF THE
DEFECT, SO FOR EXAMPLE,
GROOMING, BEFORE THE TAMOXIFEN,
YOU SKIN OFF, AFTER THE
TAMOXIFEN FEEDING, THE LESION
HEALED THE ONLY DIFFERENCE THE
AIR GROWS BACK WHITE, WE THINK
IT'S RELATED TO TAMOXIFEN, IT'S
A BREAST CANCER DRUG, CAN YOU
SEE SOME OF THE PATIENTS ONLINE,
SOME PATIENTS, OH WHY MY HAIR
BECOME WHITE?
SO YOU CAN SEE THAT THIS CAN BE
REVERSED.
WE ALSO FOUND SURPRISING SOCIAL
INTERACTION ALSO CAN BE
REVERSED, HOWEVER, MANY THINGS
WE CANNOT REVERSE, ANXIETY LIKE
BEHAVIOR, SENSORY
OVERSENSITIVITY, MOTOR DEFECT,
NONE OF THEM WE CAN REVERSE IN
ADULT.
HOW IF YOU TURN ON SHANK 3 GENE,
3 WEEKS POSTNATAL P21 OR 2 WEEKS
YOU CAN REVERSE ALMOST
EVERYTHING, SO THAT -- THERE
MIGHT BE I -- THIS IS CONSISTENT
WITH ALL THE GREATER WORK DONE
BY VIDEO SYSTEM, WHEN DO YOU
CRASS THE CRITICAL PERIOD,
WHETHER IT'S CRITICAL LANGUAGE
OR OTHER THINGS ARE HARDER TO
MODIFY, BUT AT LEAST THIS TELLS,
THERE MIGHT BE A POSTNATAL
WINDOW FOR GENE THERAPY IF YOU
CAN TURN OFF THE GENE WITHIN A
CERTAIN PERIOD OF TIME
POSTINATEALLY, YOU MIGHT BE
CORRECT MOST OF THE THINGS, SO
OF COURSE, FOR HUMAN PATIENT WE
CANNOT USE THIS GENETIC
APPROACH, BUT WE CAN TRY GENE
THERAPY.
SO WE DECIDE TO USE JOON
REPLACEMENT GENE THERAPY, THE
REASON IS MOST PATIENT, AT LEAST
MANY, MANY PATIENT RIGHT NOW
WITH THE GENOME TECHNOLOGY WHICH
HAVE GREAT PROMISES FOR MEDICINE
CANNOT DO THIS EASILY.
AND ALSO, EVEN THE POINT OF
MUTATIONS OVER THE GENE, LIKE
ISSUE MUTATION, THERE'S NOT A
SINGLE MEDIAN ACCOUNT FOR MANY
PATIENTS OR MOST PATIENTS, SO WE
DECIDE WE WANT A SYSTEM THAT
CAN, YOU KNOW TREAT FOR, YOU
KNOW VAST MAJORITY OF THE
PATIENT, AND SO, WE DECIDE TO
USE GENE REPLACEMENT
POSTINATEALLY, BUT RIGHT NOW,
THE MAIN WAY TO DELIVER INTO
GENES INTO THE BRAIN IS USE AAV,
BUT AAV HAS A PACKAGING LIMIT SO
SHANK 3 IS TOO BIG, WAY TOO BIG
FOR AAV SO WE DECIDE TO MAKE A
MINISHAIRCHG 3 WHICH CONTAINS
SOME OF THE MOST IMPORTANT
DOMAINS AND STILL FUNCTIONAL.
SO WE TESTED INVITRO, WE TEST IT
IN VIVO USING PHYSIOLOGY AND
COLOCALIZATION SHOW IT'S STILL
FUNCTIONAL AND THE USE OF BI ONY
CHEMISTRY TO SHOW IT GOES TO
PSD.
SO WITH THIS APPROACH, WE
ACTUALLY USE THE AAV CAPSID THAT
IT CAN PASS BBB, SO CAN YOU DO
IV INJECTION DEVELOPED BY VIVIAN
GRANAD O OUT OF CALTECH, AND
BAND IS NOW AT THE BROAD
INSWORKING WITH COLLEAGUES NOW,
SO WE CAN USE THIS BECAUSE OF
GENE THERAPY WE CAN INJECT AT
DIFFERENT TIME POINT.
POSTINATEAAL 0, P47, P28, AND
THEN YOU WAIT FOR 2 MONTHS AND
YOU DO BIOCHEMICAL TESTS, CAN
YOU DO ELECTROPHYSIOLOGY TESTS
AND YOU CAN DO BEHAVIOR TEST TO
SEE, AT WHICH TIME POINT CAN YOU
RESCUE THEM, SECOND AT WHICH
TIME POINT YOU RESCUE THEM, WHAT
WE FOUND IN ALL THESE TIME
POINT, YOU CAN ALMOST RESCUE,
ALMOST ALL THE PHENOTYPES, VERY
CLEARLY.
SO THIS IS EXAMPLE, SO CORTICALE
SYNAPTIC FUNCTION, THIS IS A
FIELD RECORDING, BLACK IS THE
WILD-TYPE, RED IS THE SHANK 3
INJECTED CAN AAVG, AND THEN THE
BLUE IS THE INJECTED WITH,A,A V
MINISHANK SO CAN YOU SEE THAT
CORTICALE STRATEGIC PLANS ON HAS
FUNCTION, THERE'S NO PREFERENCE
FOR SOCIAL COMPARED TO THE
OBJECT, BUT THE GENE THERAPY
RESCUED THE BEHAVIOR, SO THAT
GAVE US SOME HOPE THAT MAYBE
THIS GENE THERAPY WILL WORK, THE
ISSUE FOR US AT THIS TIME IS TO
MOVE THIS TO HUMANS IS A REAL
CHALLENGE BECAUSE GENE THERAPY
UNLIKE DRUG TESTING.
IF YOU TAKE A DRUG, IT HAS A
SIDE EFFECT, A SEVERE SIDE
EFFECT OR NOT EFFECTIVE, YOU CAN
TOP IT WITH AAV, YOU INJECT IT,
YOU CAN NEVER TAKE IT OUT, SO WE
PROBABLY NEED BETTER MODEL OR AT
LEAST ANOTHER MODEL CLOSER TO
HUMAN TO TEST THEM.
SO WE ALL KNOW MICE ARE GREAT
MODELS FOR BIOMEDICAL RESEARCH,
IT'S EVOLUTIONAL FOR THE PAST
MANY, MANY DECADES BUT THERE ARE
ALSO SIGNIFICANT
DIFFERENCESENTIOUS SPECIALLY NOW
WITH, YOU KNOW, ANATOMICAL
STUDIES, CONNECTIVITY STUDIES
AND A SINGLE CELL STUDIES AND
CELL TYPE OF STUDIES THERE ARE
STILL MANY DIFFERENCES, SO IT
WOULD BE REALLY GOOD FOR US,
ESPECIALLY TO STUDY SOCIAL
COGNITION OR OTHER HIGH
FUNCTIONING, WE HAVE NONHUMAN
PRIMATE MODEL SO THAT TIME WAS
MANY, MANY DONORS AND NOW MORE
RECENTLY WITH THE BRAIN
INITIATIVE AND WE ARE REALLY,
REALLY GRATEFUL FOR THE BRAIN
INITIATIVE ESPECIALENTIALLY AND
WE GENERATE MARMOSET MODEL.
IT'S ANOTHER SMALL NONHUMAN
PRIMATE, A LITTLE MONKEY,
THEY'RE NOT AS CLOSE AS MACAQUES
TO THE HUMANS BUT THEY'RE VERY
SMALL, THEIR REPRODUCTION IS
MUCH FASTER, IT'S ACTUALLY
MATURE IN A ABOUT A YEAR AND A
HELPFUL AND THEY GIVE BIRTH
TWICE A YEAR, THEY EACH GIVE
BIRTH TO TWIN OR TRIPLET, SO IF
YOU WANT A ANIMAL, YOU WANT TO
BREED FAST SO YOU DON'T HAVE TO
WEIGHT FOR 20 YEARS, HAVE YOU A
COLONY YOU CAN STUDY THEM.
SO OUR FIRST MODEL WAS ACTUALLY
SHANK 3 WE MADE A SMALL
CONDITION, HUMAN CONDITION EXON
21 AND WE MADE A SINGLE JEAN
INSERTION WHICH IS COMMONLY
FOUND IN SEVERAL ASD PATIENTS
NOW, THIS IS MOST COMMON IN
SEARCHING FOR SHANK 3 MUTATION
IN PATIENT.
AND SO NOW WE HAVE A SIGNIFICANT
NUMBER OF F1s, THE PURE
HETEROZYGOUS, SO IN MARMOSET WE
STUDY CONDITION, SO THEN THE
OTHER ADVANTAGE IS MARMOSET IS
VERY SMALL ISSUES THE KAINL IS
VERY BIG COMPARED TO BODY SIZE
AND THEY LIVE WITH FAMILY AND
YOU CAN SEE WE DEVELOP A MACHINE
BASED BEHAVIOR TRACKING AND WE
CAN TRACK THEM REALLY WELL, NOW
YOU CAN JUST PUT A CAMERA ON,
YOU CAN TRACK THEM FOR DAYS OR
WEEKS OR MONTH BECAUSE IT'S
AUTOMATED BEHAVIOR
QUANTIFICATION, YOU DON'T HAVE
TO SPEND A LOT OF TIME DOING ALL
THE WORK FOR YOU, RIGHT?
SO I WON'T JUST GIVE A COUPLE
EXAMPLES HOW WE STUDY THEM AND
WHAT DO THEY LOOK LIKE, SO WITH
THIS KIND OF PROGRAM, CAN YOU
TRACK THEIR BEHAVIOR, SO AGAIN,
THERE ARE 2 MARMOSET IN THE HOME
CAGE, ALL THESE STUDIES ARE IN
HOME CAGE.
YOU CAN LOOK AT IT, THEY ARE
FRACTION OF TIME, THEY STAY
TOGETHER, WHICH IS SOCIAL, THEY
GROMEACH OTHER, THEY AT A AND
LOOKING TOGETHER, YOU KNOW LIKE
COORDINATED LOOKING AND JUST
SITTING THERE TO BE FED.
AND ALSO FRACTION TIME THEY ARE
APART FROM EACH OTHER, THEY ARE
RUNNING AROUND, DOING THEIR OWN
TYPE.
AND KIELD TYPE, IF THE 2 ARE
WILD-TYPE, WHETHER IT'S YOUNG
ANIMAL OR ADULT ANIMAL THEY ALSO
HAVE 2 PEAKS, NEAR FROM EACH
OTHER, APART FROM EACH OTHER,
BUT IF IT WAS WITH THE SHANK 3,
WE SEE IS NEAR EACH OTHER, PEAK
IS GONE.
IT'S ALMOST LIKE YOU GO TO A
BAR, YOU WANT TO MEET SOMEONE,
YOU GOTTA DRINK, YOU SIT DOWN
WITH SOMEONE, YOU START TO
TALKING, SOCIAL, IF THAT PERSON
NEVER RESPOND TO YOU, YOU WILL
NOT STAY THERE VERY LONG, YOU
WILL FIND SOMEWHERE ELSE SO IT'S
AN INTERESTING PHENOMENA, YOU
CAN SEE FROM THIS AUTOMATED
BEHAVIOR TRACKING, THERE ARE
MANY OTHER THINGS CAN YOU DO FOR
EXAMPLE, MARMOSET LOVE iPAD,
BUT THEY LOVE iPAD, BUT THEY
LOVE TOUCH PAD, SO THEY ALL GO
IN FRONT.
AND YOU TEACH THEM LEARN, TOUCH
A PARTICULAR SPOT, YOU GET A
REWARD, THEY LOVE MARSHMALLOW,
SO WE GIVE THEM A LITTLE TINY
TINY MARSHMALLOW, AND LET THEM
GO AND TOUCH IT AGAIN.
SO CAN YOU SEE WHEN WE HAVE A
TOUCH PAD IS THE BLUE SQUARE,
THEY ALL STAY IN FRONT WHETHER
IT'S WILD-TYPE OR SHANK 3.
IF YOU CHANGE THIS BLUE SQUARE
TO A MARMOSET IMAGING WILD-TYPE
DOESN'T CARE, MUTANT THEY STAY
BACK MORE, AND THEY STILL WILL
GO GET REWARD BUT THEY DON'T
WANT TO AT A THERE ALL THE TIME
THEY JUST GO THERE SO IT'S A
REALLY INTERESTING PHENOMENA
EVEN THE FACE OF MARMOSET, THEY
DON'T LIKE TO STAY THERE ALL THE
TIME.
AND YOU CAN ALSO TEST IT WITH
ANIMAL, RIGHT?
SO THIS IS SMALL CAGE, PUT IT
INSIDE THE CAGE BECAUSE THEY
CANNOT COME OUT, BUT OH, THE
WILD-TYPE, YOU SEE WILD-TYPE, OH
COME TO LOOK, WHO IS THE NEW
KIDOT BLOCK JUST LIKE HUMAN
BEHAVIOR, AND YOU WANT TO
INVESTIGATE, BUT IF IT'S A
MUTANT MARMOSET, THEY ARE
ACTUALLY REALLY SCARED OF
STRANGER MARMOSETS THEY WILL
SCREAM AND CLIMB ON TOP OF THE
CAGE.
THAT'S USUALLY THEIR SAFE PLACE,
UNLIKE TREES, SO YOU SEE THERE
IS A SIGNIFICANT RESPONSE TO NEW
THINGS INCLUDING NEW NARMA SET.
SO THIS KIND OF ASSAY WE CAN DO
A LOT THIJS, THE RISK OF
BEHOUSTON AND RICE FOOTBALLS
BECAUSE THEY'RE IN THE HOME
CAGE, YOU CAN DO iPAD,
MOTIVATION, SOCIAL, AND YOU CAN
DO A LOT OF THINGS.
AND THEN YOU CAN INDUCE THE
BEHAVIOR, BECAUSE THE MARMOSET
ARE VERY HETEROGENEOUS, THEY ARE
NOT INBRED, THEY ARE COMPLETELY
OUTBRED, WE DO EVERYTHING TO
MAKE SURE THEY ARE NOT INBRED,
SO THE BEHAVIOR IS VERY VARIABLE
JUST LIKE HUMANS BUT WE FOUND
THAT IF YOU DO COMPARE BEFORE
AND AFTER STIMULI, THAT'S THE
MOST ROBUST PHENOTYPE YOU CAN
FIND SO THIS IS ACTUALLY 1 OF
THE MOST IMPORTANT ASSAYS WE DO,
AND YOU CAN DO BASIC FUNCTION
WHETHER IT'S FEEDING, GAIT OR
SLEEPING TIME ALL THE THINGS CAN
YOU DO SO WITH THIS KIND OF
ASSAY, NOW WE CAN TEST WILL YOUR
GENE THERAPY IMPROVE ANY OF
THESE ISSUES.
RIGHT?
SO WE DID THE 2 GROUP GENE
THERAPY.
WE HAVE A LARGE WILD-TYPE
CONTROL WE TRACK THEM FOR LONG,
LONG TIME, SO WE KNOW THE BASE
LINE, AND THEN WE HAVE A GROUP
OF SHANK 3, WITHOUT GENE THERAPY
AND 1 GROUP OF SHANK 3 STARTED
WITH 9 MONTHS, 9 MONTH SYSTEM
ABOUT 10-12 YEAR-OLD KIDS IN
DEVELOPMENT AND THEN WE HAVE
ANOTHER GROUP STARTED WITH 3
MONTHS OLDER, BABY, SO 3 MONTHS
OLDER, ABOUT 3-4 YEARS OLD HUMAN
BABY.
AND THEN WE WAIT FOR 3 MONTHS,
THIS IS THE IV INJECTION, WE
FOUND THAT WITH THE SEPARATE
MARMOSET, WITH IV INJECTION WITH
THIS PARTICULAR CAPSID DEVELOPER
IN COLLABORATION, MOSTLY BY LAB
AT BROAD RIGHT NOW, THIS IS
SPECIFIC TO MARMOSET
UNFORTUNATELY, IT DOESN'T GO
THROUGH EVEN MACAQUES, SO I
DON'T THINK IT WILL WORK WITH
HUMANS BUT IT WORKS REALLY WELL
WITH MARMOSET AND YOU CAN IN
FACT USE THE PROMOTER, AND
AFFECT ABOUT 15-20% NEURONS IN
THE BRAIN WITH IV INJECTION.
SO WE WAITED IF ARE 3 MONTHS AND
THEN EVERY 3 MONTHS WE TESTED
THEIR BEHAVIOR AND WHAT WE FOUND
IS, WE FOUND HOW DO YOU --
BECAUSE YOU CANNOT USE 20 --
LIKE MICE, 20 MONKEYS FOR
WILD-TYPE, 20 MONKEYS ARE FOR
GENE THERAPY, SO WE ONLY HAVE 8
MONKEYS, RIGHT, FOR THE GENE
THERAPY, 4 FROM 3 MONTHS, 4 FROM
9 MONTHS, SO WE FOUND THE MOST
ROBUST WAY IS TO COMPARE THEIR
BEHAVIOR TO THEIR CAGE MATE,
WILD-TYPE CAGE MATE BECAUSE
MONKEYS LIVING IN DIFFERENT
ROOMS, DIFFERENT CAGES, IF 1
CAGE MATE IS VERY HYPER AFFECT,
IT WILL AFFECT THE OTHERS, THEY
WILL GET MORE ACTIVE.
AND THEN, IF SOMEONE COME TO
CHANGE THE CAGE, MAKE A NOTICES,
THE WHOLE ROOM WILL BE AFFECTED.
BUT OTHER ROOM WILL NOT BE
AFFECTED.
SO WE FOUND IT'S MOST ROBUST WAY
IS TO SEE THE MONKEY'S BEHAVIOR,
HOW CORRELATED WAS THE WILD-TYPE
CONTROL IN THE SAME CAGE.
SO THESE 2 IN THE SAME CAGE FYOU
LOOK AT BEHAVIOR, VERY LITTLE
DIFFERENCE, WILD-TYPE.
BUT IN THE SAME CAGE IF 1 IS
WILD-TYPE AND 1 IS SHANK 3, CAN
YOU SEE, THEY'RE VERY DIFFERENT,
SO NOW YOU CAN SEE, CAN YOU
REVERSE THIS INTO BECOME LIKE
THIS, RIGHT?
SO WE DID A LOT OF ASSAYS,
TRACKING MANY, MANY HOURS, THIS
IS 5 SOLID SESSIONS ALMOST LIKE
200 QUESTIONS OR NUMBER OF
QUESTIONS AND YOU CLICK ALL THE
THINGS, WHAT WE FOUND IS THIS IS
WILD-TYPE, RIGHT, THEY HAVE
COORDINATION, THEY HAVE
CORRELATION WITH THE WILD-TYPE
CAGE MATE THIS, IS SHANK 3, WAY
LESS CORRELATED BEHAVIOR WITH
THE WILD-TYPE AND GENE THERAPY
AND ACTIVITIES AND PROJECTS
OVERALL YOU CAN IT CORRECTED TO
THE WILD-TYPE LEVEL.
THEN YOU GO AND LOOK AT EACH
INDIVIDUAL BEHAVIOR, WHICH IS
CORRECTED WHICH IS NOT
CORRECTED, SO FROM BEHAVIOR
POINT OF VIEW, YOU CAN SEE FOR
WILD-TYPE, THEY ARE WIDER
SPREAD, FOR MONKEYS WIDER
SPREAD, BUT YOU CAN SEE AVERAGE
ANIMAL THIS, IS TOUCH SCREEN,
RIGHT IN AND THEY ARE LESS
CORRELATE WIDE COMPARED TO
WILD-TYPE, GENE THR 'Y CAN
RESTORE TO WILD-TYPE LEVEL, THIS
IS MA REMEMBERRA SET FACE, SAME
THING GENE THERAPY CAN CORRECT
ON AVERAGE AND THIS IS FOR NOVEL
TOY THIS, IS FOR STRANGER
MARMOSET, YOU SEE THEY CAN ALL
CORRECT THROUGH WILD-TYPE LEVEL
EMPLOY NOW WITH THIS, WE CAN --
IN MICE WE TRY VERY HARD AND
MANY OTHER LEVELS, MANY OTHER
LAB TRIED TO SEE -- WHETHER YOU
SEE A LEARNING DEFECT IN
MARMOSET, IT WAS HARD TO FIND.
IN OUR ORIGINAL PUBLICATION, WE
SAID OTHER THINGS WE DIDN'T SEE
HOMOZYGOUS, WE DID NOT SEE
LEARNING DEFECT, THAT WAS
PUBLISHED IN OUR PAPER, AND BUT
IN MARMOSET, IT'S HETEROGENEOUS
OR O ZYGOUS, IN HUMANS VERY
SEVERE PROBLEM, THEIR AVERAGE IQ
IS HARD TO MEASURE, 80 OR 90%
PEOPLE YOU CAN'T, BUT THE 1S YOU
CAN MEASURE, THE AVERAGE IQ IS
ABOUT 30 OR 40.
SO IT'S VERY LOW.
SO FOR MARMOSET, WE CAN TEST
WHETHER WE SEE A LEARNING
DEFECT, SO WHAT WE FOUND IS THAT
YOU CAN USE MANY COGNITIVE
TESTING YOU CAN DO LEARNING
DEFECT, SO THIS IS A VERY
SIMPLE, RIGHT?
BASICALLY, YOU KNOW 2-POINT
CHOICE, SO BASICALLY YOU TOUCH
RIGHT PLACE, YOU GET NEW 1, YOU
CAN SEE VERY QUICKLY, WILD-TYPE
MAKE VERY FEW MISTAKES BUT FOR
SHANK 3 WE FOUND 2 PHENOMENA.
ONE IS THEY INSIST TO TOUCH 1
SPOT, LIKE THE MARMOSET WILL
JUST GO THERE AND TOUCH 1 SPOT.
AND SO THEY HAVE -- YOU KNOW
THIS IS JUST 1 -- SO THEN WE
FORCE THEM TO TOUCH MORE, LIKE
IF YOU KEEP THEM SAME PLACE, YOU
DON'T GET REWARD, SO EVEN WITH
THAT, YOU STILL HAVE PREFERENCE
AND LIKE WILD-TYPE, AND THEY
ALSO MAKE A LOT OF MISTAKES, SO
THEY'RE MUCH SLOWER AND ALONG
THE WAY THEY MAKE MISTAKES.
SO CAN YOU VISUALIZE THIS,
BASICALLY THIS IS TOUCH PAD,
THEY HAVE TO TOUCH THIS SPOT TO
GET THE REWARD, WILD-TYPE DO
REALLY WELL, EVERYTHING IN THE
MIDDLE AND THEY TOUCH MANY, MANY
TIMES.
SHANK 3, THEY KNOW THEY NEED TO
TOUCH BUT THEY TOUCH THE SCREEN,
THEY DON'T UNDERSTAND YOU HAVE
TO TOUCH IT HERE AND YOU HAVE TO
TOUCH IT LESS TIMES SO LESS
TRIALS BUT WITH GENE THERAPY,
YOU MOVE ALMOST MOST OF THIS
INTO THE MITSD SO THAT YOU CAN
CORRECT THIS KIND OF DEFECT, SO
TO US, EVEN MORE INTERESTING TO
LOOK AT SEX LEVEL CHANGES
BECAUSE IF YOU CAN PERMEATE THE
SECTION, YOU CAN REDICT THE
REWARD, THIS IS RESTING STATE
FUNCTIONAL MRI, SO IF YOU
COMPARE THE LOCAL FUNCTIONAL
CONNECTIVITY AND YOU CAN SEE
THIS IS SUBTRACTION, WILD-TYPE
FROM THE SHANK 3, RED MEANS
HIGHER, BLUE MEANS MUTANT IS
HIGHER, SO IN GENERAL WE HAVE
FEELING FRONT CORTEX ACTIVITY IN
THE SHANK 3 MARMOSET IS LOWER IN
SHANK 3 BUT THE OTHER PART IS
HIGHER IN SHANK 3, BUT IF YOU DO
STATISTICAL ANALYSIS, THE STEADY
SIGNIFICANCE IS PART OF THE
REDUCED ACTIVITY AND REDUCED
FUNCTIONAL ACTIVITY IS VERY
SIGNIFICANT, AND SO YOU CAN USE
DIFFERENT VIEWS AND THIS WAS
INCOLLABORATION WITH FRED AND
BOB'S LAB, AND WITH THE GENE
THERAPY, YOU CAN SEE, THIS IS --
I JUST SHOW YOU IF YOU COMPARE
WILD-TYPE WITH GENE THERAPY,
SHANK 3 GENE THERAPY, YOU SEE
THE DIFFERENCE IS GONE.
SO CAN YOU SEE THAT NOT ONLY IT
CAN CHANGE THE CORRECTED
BEHAVIOR, BUT IT CAN ALSO
CORRECT THE RESTING STATE
FUNCTIONAL CONNECTIVITY IN THIS
ANNUAL, WHETHER IT'S LOCAL
FUNCTIONAL CONNECTIVITY OR
GLOBAL FUNCTIONAL CONNECTIVITY.
SO YOU CAN SEE, SO WE'RE REALLY
EXCITED ABOUT THIS KIND -- SO
WITH THIS POST MOUSE AND THE
MONKEY STUDY, THIS THING IS NOT
IN CLINICAL TRIAL, ACTUALLY,
THEY JUST DOSED FIRST PATIENT IN
FEBRUARY, SO WE DON'T KNOW
RESULT AND THE HOPE IS THIS WILL
REALLY HELP PATIENT IN THE LONG
RUN.
THIS IS THE FIRST TRY OF THE
GENE THERAPY FOR THE SHANK 3 AND
WE'RE HOPING WITH THE CLINICAL
FEEDBACK, IT IS EFFECTIVE OR
HAVE SOME EFFECT AND THEN WE CAN
CONTINUE TO IMPROVE THIS TYPE
BECAUSE THESE PATIENTS ARE VERY
SEVERELY DISABLED, NONE OF THEM
CAN REALLY LIVE INDEPENDENTLY.
SO, IN MY VIEW, MANY OF THESE
MONOGENIC DISORDERS, WE CAN
DEVELOP DIFFERENT GENE THERAPY
APPROACHES IN THE FUTURE AND THE
MOST DIFFICULT PART IS HOW TO
GET A NEW VECTOR THAT YOU CAN IV
INJECTION AND GOOD BRAIN
COVERAGE WHICH NOW SEVERAL LABS
AND COMPANY MADE PROGRESS.
THE OTHER IS THIS IS RARE
GENETIC DISORDERS, MOST PHARMA
WOULD -- HOW TO GET SOCIETY TO
INVEST IN THIS RARE GENERATEDET
INCREASE IN BODY DISORDER
BECAUSE THIS IS 1 OF THE FEW
CASES WE CAN REALLY MAKE
PROGRESS.
SO WE LEARN A LOT FROM THIS
MARMOSET SHANK 3 MODEL SO WE
STARTED THINKING WHAT ELSE WE
CAN DO TO HELP THE RESEARCH
COMMUNITY AND THE BIOTECH
COMMUNITY TO GENERATE OTHER
MODELS THAT HELP US TEST THE
THERAPEUTICS IN THESE MODELS.
SO NOW WE GENERATED MECP2 MODELS
WHICH IS THE HUMAN R-GENINE,
STOP PROGRAM, THIS IS ALSO A
TERRIBLE DISORDER AFFECT ALMOST
EXCLUSIVELY GIRLS BECAUSE I
THINK BOYS CANNOT SURVIVE, AND
SO, THERE ARE MANY GENE
THERAPIES, FOR RETT SYNDROME, SO
HOPEFULLY WE GENERATE THIS MODEL
TO BE USEFUL IN THE FUTURE.
WE ALSO GENERATED A VERY ONSET
MUSEUM TAITIONZ, P117S AND P117
L MUTATION, THIS PATIENT BECOME
VERY SEVERE AT 30 YEARS OLD AND
SO 30 YEARS, OR ABOUT 4 YEARS
OLD MARNLA SET, SO YOU DON'T
HAVE TO WAIT 20 YEARS TO STUDY
THIS.
SO WE'RE HOPING THIS WILL GIVE
US SOME INSIGHT.
-- WAS HERE BUT ALSO AT
PITTSBURGH ALSO MADE DIFFERENT
MUTATIONS WHICH IS LATER ONSET,
I THINK IS ALREADY SHOWED THERE
ARE PLAQUES IN THIS MARMOSET, SO
USUALLY KNOCKIN OPPOSING
MUTATION IN MICE SO YOU DON'T
SEE THESE PLAQUE CHANGES.
SO WE ARE HOPEFUL.
WITH THE STANLEY CONSORTIUM OF
THE GLOBAL CONSORTIUM OF THE
SCHIZOPHRENIA WHOLE EXOHM
SEQUENCING THIS IS THE FIRST
PUBLICATION FROM STANLEY CENTER
FROM THE GLOBAL CONSORTIUM
IDENTIFY DEN HIGH PENETRANT RARE
GENETIC MUTATIONS CONTRIBUTED TO
SKITS SKITS
SCHIZOPHRENIA AND WE TESTED
INITIALLY GENERATED A FEW MOUSE
MODELS TO TEST HETEROZYGOUS, YOU
SEE SOME OF THE FINE O TYPES,
SOME OF THE INTERESTING
PHENOTYPES, SO NOW WE GENERATE
FIRST MARMOSET MODEL, HOPEFULLY
THIS WILL BE USEFUL, ALL THE
GERONTOLOGYSTS MEMORY CLONE AND
ENGINEERING TECHNOLOGY WHETHER
IT'S A PRIME EDITING, GENOME
EDITING, SMALL KNOCKINGS OUT, WE
MADE IT VERY EFFECTIVE IN
MARMOSET THANKS TO THE BRAIN
INITIATIVE AND THE NIH SUPPORT
FOR GENOME ENGINEERING IN
NONHUMAN PRIMATE PROGRAM.
I SAID MOST OF THE DISORDERS IN
MENTAL HEALTH INCLUDING
SCHIZOPHRENIA, DEPRESSION,
AUTISM, THEY'RE NOT MONOGENIC SO
CAN WE USE A SUCH DYSFUNCTION AS
A BIOMARKER AND THERAPEUTIC
TESTING THAT CO POTENTIALLY
TRANSLATE INTO HELP FOR HUMANS.
SO WE ARE IDENTIFYING ON HOW DO
WE IDENTIFY CIRC YOUS THAT
BEHAVIOR TRIGGERED DEFECT, I
SHOWED IT IN MARMOSET AND PEOPLE
ALREADY PUBLISH IN MOUSE SHANK 3
MODELS, YOU CAN DO RESTING
ESTATE WHICH IS ANESTHETIZED
ANIMAL BUT IS THIS SAME OR
DIFFERENT FROM A BEHAVIOR WE'RE
WEAK TRIGGER?
BECAUSE MOST TRIGGER AUTISM IS
IN SOCIAL ENVIRONMENT OR
SOMETHING IRRITATED THEM SO CAN
WE TEST BEHAVIOR TRIGGERED
CIRCUITS ACTIVITY WHETHER THEY
HAVE A DIFFERENCE?
SO MY LAB ALWAYS FOCUSED ON
THYME O CORTICAL CIRCUITS AND
BECAUSE OF CONSERVATION BUT ALSO
BECAUSE IT'S IMPLICATED IN MANY
DISORDERS, SO WE IN THE LAST FEW
YEARS, WE ARE -- WE ARE STILL
DOING FUNCTIONAL MRI WITH ANIMAL
BUT WE ARE MORE INTERESTED
NONAPOPTOTIC YOU IN USING THIS
RELATIVELY NEW TECHNOLOGY CALLED
FUNCTIONAL ULTRASOUND IMAGING
BECAUSE IT HAS A HIGHER
RESOLUTION BUT IT ALSO CAN DO
WEAK ANIMAL, SOMETIMES CAN YOU
DO BEHAVING ANIMALS BECAUSE IT'S
A PROBE ON THE HEAD.
SO THIS IS BASICALLY SIMILAR --
IT'S BASICALLY BLOOD FLOW TOPLER
IMAGING, SIMILAR TO FUNCTIONAL
MRI, YOU LOOK AT BLOOD FLOW
REPRESENTATIVE OF THE COLORED
WITH THE NEURAL ACTIVITY.
SO WE TESTED THIS INITIALLY AN
ESTIMATE THADITIZEED RESTING
STATE BECAUSE RESTING STATE FMRI
HAS BEEN DONE MULTITELETIMES IN
SHANK 3 ANIMAL AND WE KNOW IT
WAS REPORTED WITH REDUCED
FUNCTIONAL CONNECTIVITY IN THE
FUNCTIONAL CORTEX IN THE
STRIATUM, AND WE SHOWED THAT IN
THE CORTEX OF THE MARMOSET, TOO.
SO WE SHOWED SHANK 3.
SO WE TEST IS FIND A SIMILAR
PHENOMENA IN THE SHANK 3 MICE,
THIS IS OUR 10 ANIMALS FOR EACH,
CAN YOU SEE WILD-TYPE, AND YOU
CAN SEE THAT IT'S A RELATIVE
ACTIVITY, RIGHT?
YOU CAN SEE KNOCK OUT WITH
FRONTAL CORTEXT AND STRIATUM SO
YOU CAN CONFIRM THIS NEW
APPROACH, NEW TECHNOLOGY CAN BE
USED TOY DID TECT CIRCUITS
ACTIVITY CHANGE AND MOUSE
MODELS.
SO THEN WE DESIGN EXPERIMENT TO
LOOK AT BEHAVIOR TRADING SO
BASICALLY WHAT WE DID WAS TO
MAKE THE EFFECTS OF THIS TESTING
ANIMAL THEN WE HAVE A PROBE
ANIMAL, YOU HAVE A LOOP OF TUBE,
SO THE ANIMAL CAN GO OVER SOCIAL
AND GO AWAY.
SO HAVE YOU WINDOWS SOCIAL,
NONSOCIAL, SOCIAL, NONSOCIAL SO
CAN YOU SEE WHEN THEY ARE
SOCIAL, THIS BOX ARE SOCIAL, YOU
SEE THE ACTIVITY INCREASE FROM
THE CORTEX AND IN THE STRIATUM.
SO THEN YOU SEE THE SAME THING
WITH IMK OUT ANIMAL.
TO OUR SURPRISE IN A WEAK
CONDITION, EVERY TIME THERE'S
SOCIAL ACTIVITY, THE FRONT
CORTEX AND STRIATAL ACTIVITY
DRAMATICALLY INCREASE, SO
PREVIOUSLY WE WERE THINKING
BASED ON ELECTROPHYSIOLOGY IN
SLICE, BASED ON FMRI RESTING
STATE WE NEED TO BOOST THE
CORTICALE SYNAPSE, NOT WITH
THIS, BUT WHAT WE NEED ACTUALLY
IS TO CALM DOWN THE CORTICALE
STRIATAL FUNCTION SO THIS IS NOT
A 1 IMAGING, THIS IS THE AVERAGE
IMAGING FROM ABOUT 10 ANIMALS,
CAN YOU SEE THE SIGNIFICANT
ESPECIALLY IN THE FRONTAL CORTEX
AND STRIATUM, SO THIS HAS BECOME
A NEW KIND OF ANGLE TO NOT ONLY
LOOK AT THE CIRCUIT BUT ALSO TO
LOOK AT THE WHAT KIND OF CELL
TYPE CHANGED BUT MORE IMPORTANT
TO US, CAN WE FIND THE TARGET
CALM DOWN THE CIRCUITS OF THIS
BEHAVIOR TRIGGERED CIRCUITS
NEURONAL HYPER-ACTIVITY, IF WE
CAN CORRECT THAT, MAYBE WE WILL
ELIMINATE IT FOR MOST OF THIS
BEHAVIOR DEFECT.
SO WE FOUND THE TARGET AND WE
FOUND THE MODULATE BECAUSE THE
EXPRESSION PATTERNS AND THEY ARE
FUNCTIONING IN GABBAERGIC
NEURONS ACTUALLY SO WHAT WE
FOUND IS THAT WILD-TYPE, RIGHT,
SO CAN YOU SEE THERE'S BEFORE
DRUG KNOCK OUT BEFORE DRUG, YOU
HAVE HYPER ACTIVE.
THESE ARE SOCIAL TRIGGERED
NEUROSUBACTIVITY.
WE GIVE 1 DOSE, 1 INJECTION, 2
HOURS LATER WILD-TYPE HAVE
LITTLE BIT ACTIVITY, BUT NORMAL
ALMOST KNOCK OUT, 2 DOSE, 7 DAYS
LATER IT STILL HAS EFFECT.
IT START TO CREEP BACK BUT IT
STILL HAS IT, IT'S VERY SIMILAR
TO KETA MEAN -- IT'S A LONG
LASTING MODULATOR ACTIVITY, AND
SO THE PREDICTION, IF YOU CAN
CORRECT THIS SO WELL AT A LEVEL,
YOU SHOULD BE ABLE TO PREDICT
THE BEHAVIOR WOULD CHANGE, THIS
IS TRIGGERED BY SOCIAL
INTERACTION SO WE DID TEST THEM
USING THE SAME SOCIAL ASSAY.
ONE DOSE, THEN YOU TEST 24 HOURS
LATER AND 21 DAYS LATER.
WE ALSO TESTED 7.
SO CAN YOU SEE THAT IT VERY
CLEARLY TO CHANGE THE BEHAVIOR
THAT YOU CAN RESTORE THE SOCIAL
INTERACTION VERY CLEAR, THIS IS
BLACK WILD-TYPE WITH VEHICLE,
CAN DRUG MUTANT WITH VEHICLE,
WITH DRUG, YOU CAN SEE VERY
CLEARLY, IT CAN LAST VERY LONG,
21 DAYS LATER STILL HAS EFFECT,
SIMILARLY FOR GROOMING BEHAVIOR,
1 DOSE, TOP LINE, OPEN CIRCLE IS
KNOCK OUT, WITH VEHICLE.
THE BLACK OPEN CIRCLE IS THE
WILD-TYPE WITH VEHICLE.
THE FILLED CIRCLE IS THE KNOCK
OUT WITH THE NR2 B ANTAG NAST
AND THE WHITE CIRC CELL
WILD-TYPE WITH DRUG, SO YOU CAN
SEE IT LASTS ALMOST 4 WEEKS SO
WE'RE REALLY EXCITED ABOUT THIS
LONG LASTING EFFECT AND WE TRY
TO FIGURE OUT WHAT IS THE
MECHANISM, ALTHOUGH A DRUG IS A
DRUG, NO COMPOUND IS A HUNDRED
PERCENT, YOU KNOW SPECIFIC.
SO WE WANT TO SEE, IS THIS
REALLY NR2 B, SO WE MAY USE
CRISPR, WE MADE A KNOCK OUT,
JUST ADULT KNOCK OUT IN THE
SHANK 3 AND IN THE ANIMAL
DIRECTLY.
SO THIS IS CONTROL WITH THE
CONTROL GUIDE RNA, SCRAMLE RNA
AS A KNOCK OUT, YOU CAN SEE WITH
THE SOCIAL INTERACTION, THE
TRIGGERS HYPER-ACTIVITY IN THE
FRONT CORTEX IN THE TRIAT UMKC,
AND WHEN YOU KNOCK OUT OF SHANK
3 AND R2 B, YOU DON'T SEE THIS
TYPE OF ACTIVITY.
SO WE CONFIRM THE NR2 B IS
REALLY THE TARGET, SO NOW, YOU
ALL KNOW BLOCK NMD RECEPTOR IS
PROBABLY NOT BEST THING TO DO
FOR KIDS AND ADULT, FIRST OF ALL
THEY ARE CLINICAL STUDIES THAT
SHOW THEY DISASSOCIATED FACT,
WITH PROBLEMS, HYPER-ACTIVITY IN
ANIMALS AT LEAST N. HUMANS IT
AWZ DISASSOCIATION EFFECT.
SO WE'RE NOW VERY INTERESTED IN,
CAN WE FIND THE DOWN STREAM
SIGNAL AS A NEW DRUG OF TARGETS
THAT COULD COME DOWN, AND THESE
HYPER-ACTIVITY BEHAVIOR
TRIGGERED TO HYPER-ACTIVITY,
WE'RE ALSO REALLY INTERESTED,
HOW THIS THING CAN WORK SO LONG
FROM A DRUG IS GONE WITHIN 24
HOURS, SO WHY DOES THE EFFECT
LAST SO LONG?
IS IT ALTP?
OR SOMETHING ELSE?
WE WANT TO KNOW, WE DON'T KNOW
YET, WE HAVE A HINT BUT WE DON'T
KNOW YET.
SO OUR HOPE IS THAT IN NEXT
COUPLE YEARS WE COULD FIGURE OUT
A TARGET AND THIS COULD BE MOVE
ON TO TESTING AND THE MONKEYS
AND HUMANS IN THE FUTURE.
SO I THINK I WILL STOP HERE, I
WANT TO ACKNOWLEDGE MANY PEOPLE,
ESPECIALLY OUR MARMOSET TEAM.
THIS IS NOT MY LAB'S PROGRAM FOR
MARMOSET, IT'S IMPOSSIBLE TO DO,
WE HAVE MANY, MANY COLLABORATORS
ISSUES IT'S A WHOLE COMMUNITY
EFFORT ESPECIALLY BOB, AND FENG,
AND STEVE, AND FENNA, SHE WAS A
POST DOC WITH STEVE Mc CARROLL
ASSISTANT PROFESSOR AT PRINCETON
UNIVERSITY, AND HARVARD, AND
THEN SPECIALLY BROAD AND MIT
ANIMAL FACILITY.
THEY TAKE CARE OF THESE ANIMALS
SO WELL, AND EVERY SURGERY YOU
NEED ANESTHESIOLOGIST OR MRI
SCANNING, ANESTHESIOLOGIST,
SURGEON, SURGEON ASSISTANT, IT'S
BETTER ARRANGED THAN YOU GO TO
HOSPITAL SURGERY, I HAD SEVERAL
SURGERY IN THE HOSPITAL REALLY
WELL DO AND THEY DO PHENOMENAL
WORK TO TAKE CARE OF THESE
ANIMALS AND WE HAVE COLLABORATE
WITH JIM PICTEL, HERE AND
AFONSO, SILVA USED TO BE HERE
AND WE HAVE OUR COLLABORATOR IN
JAPAN, PROFESSOR OKANO AND
SASAKI, THEY ARE MARMOSET FOR
ALMOST 50 YEARS, AND I ASK THEM
WHY 50 YEARS AGO THEY THINK
MARMOSET WAS GOOD AND ASK THE
PRESIDENT OF CIEA.
SO THEY TAUGHT US A LOT.
WE HAVE 3 OR 4 PEOPLE THERE THAT
ARE THE LEARNING GROUP
BIOLOGISTS AND THEY ARE OUR
LONG-TERM COLLABORATORS AND ALSO
WE HAVE COLLABORATORS IN CHINA
SOME OF THEM MY FORMER GRADUATE
STUDENT AND THEY WORK ON MACAQUE
MONKEYS AND WE ALSO LEARNED A
LOT OF BIOLOGY OF MACAQUE
MONKEYS TO HELP US ESTABLISH
GENETIC ENGINEERING AND PLATFORM
AT MIT.
AND MOST IMPORTANTLY, I REALLY
WANT TO THANK THE MANY, MANY
TALENTED AND DEDICATED ANIMAL
CARES, EACH MARMOSET AND LIKE
MICE, YOU NEED A LOT OF CARE,
ALL THE TIME AND WE CHECK THEIR
BUS ON THE CELL PHONE, WHETHER
IT'S MITTED OF THE NIGHT OR
MORNING, WE WILL GO THERE TO
MAKE SURE THE BABIES ARE IN GOOD
SHAPE SO THESE ARE VERY, VERY
DEDICATED PEOPLE AND OF COURSE
WITHOUT FINANCIAL SUPPORT NONE
OF THIS IS POSSIBLE AND REALLY I
WANT TO EMPHASIZE HOW THE NIH
BRAIN INITIATIVE MONOPOLY US AND
HELP MANY OTHERS IN NEUROSCIENCE
IN THE FIELD IN THE PAST DECADE,
THIS IS INCREDIBLE TECHNOLOGY
DEVELOPMENT AND A MODEL
DEVELOPMENT AND WE ARE REALLY
GRATEFUL TO THE SUPPORT FOR NIH
AND ALSO AGREE FOR MANY DONORS,
ESPECIALLY THE BROAD TEXT 10 FOR
RESEARCH AND ALSO MASSACHUSETTS
LIFE SCIENCE CENTER AND POITRAS
CENTER FOR PSYCHIATRIC RESEARCH
AND MANY OTHER NIMH, COMMON
FUND, ALSO NINDS AND ALSO BROAD
INSTITUTE, AND ALSO THE BRAIN
RESEARCH FOUNDATION ACTUALLY WAS
THE FIRST SMALL FOUNDATION GIVE
US MONEY TO GET US STARTED FOR
SHANK 3 MARMOSET AND MANY OTHER
DONORS, TOO, THANK YOU.
[ APPLAUSE ]
NOKAY, THANK YOU THAT WAS AN
AMAZING TALK LIKE WE ALL
EXPECTED, QUESTIONS, YES.
>> THANKS FOR AN INCREDIBLE
WORK, INSPIRING TALK THE FIRST
HAS TO DO WITH GENETIC MARMOSET
IN THE MULTIVIEW
CHARACTERIZATION OF THEIR
BEHAVIOR, DID YOU FIND THAT SOME
OF THE BEHAVIORIAL DEFICITS
ASSOCIATED WITH THE DELETION
WERE MORE RESISTANT TO OTHERS
AND THE OTHER IS A SORT OF
BROADER QUESTION, HOW DO YOU
THINK ABOUT THE PROSPECT FOR
INDIVIDUAL VARIATION AND IN
GENETICS AND ENVIRONMENT
INCLUESING HETEROGENEITY AND A
VARIABLE PEN TRANCE OF THE MIN
EPIGENETICCULATION WHEN YOU PUT
IT INTO A REAL COMMUNITY?
>> THOSE ARE VERY GOOD
QUESTIONS.
THE FIRST QUESTION IS THERE'S A
LOT OF VARIATION, NOT EVERY
ANIMAL IS RISK AT SAME LEVEL, SO
WE ARE AS A GROUP, BUT VERY
OBVIOUSLY, VERY DIFFICULT
COGNITIVE TESTING WE CANNOT
RESCUE, THERE'S ONLY A CHANCE
AND WE DON'T KNOW THAT BECAUSE
IF IT'S ENVIRONMENTAL OR YOU
NEED EARLIER OR WE DON'T KNOW
WHETHER WE HAVE NOT ENOUGH
NEURONS BECAUSE HIGH COGNITIVE
FUNCTION IS YOUR BRAIN COMPUTING
POWER.
WE ONLY GOT INTO 15, 20% OF
NEURONS YOU MAY NEED MUCH MORE
WOODER SPREAD, SO WE DON'T KNOW
BUT THERE ARE THINGS MOSTLY IS
THE COGNITIVE FUNCTION, WE
CANNOT COMPLETELY RESCUE, SIMPLE
COGNITIVE FUNCTION WE CAN
RESCUE, BUT MORE DIFFICULT 1S WE
CANNOT RESCUE COMPLETELY.
AND THE SECOND QUESTION IS ALSO
REALLY GOOD QUESTION, ALTHOUGH
WE EXACTLY GENETIC MUTATION, SO
IT'S A PHENOMENA THAT'S REALLY
INTERESTING, THEY ALL HAVE
DEFECT, BUT THEY HAVE DIFFERENT
DEFECT AND TO DIFFERENT DEGREE,
SO IF YOU CLUSTER, USE COMPUTER,
THE CLUSTER WILD-TYPE, MUTANT.
THEN YOU BRING NEW BATCHES, SAY
WILD-TYPE, WHERE'S CLUSTER, ALL
WILD-TYPE CLUSTER WERE
WILD-TYPE.
THE NEW BATCH MUTANT, BECAUSE
MUTANT IS EVERY MUTANT IS
DIFFERENT FROM WILD-TYPE BUT
EVERY MUTANT IS DIFFERENT FROM
EACH OTHER, SO YOU CAN USE THE
TRAINING TO HAVE A CLUSTER BUT
THE NEW 1 MAY NOT CLUSTER INTO,
SO THAT MEANS, SO FOR EXAMPLE,
SOME ARE VERY SOCIAL DEFICIT,
SOME ARE NOT -- SOCIAL OKAY, BUT
OTHER I THINK THISS ARE, BUT
IT'S A SPECTRUM OF THINGS AND
KIND OF SIMILAR TO HUMAN
CONDITIONS, THEY ARE NOT
IDENTICAL, EVEN IF THEY HAVE
IDENTICAL MUTATIONS, THEY ARE
JUST GENETIC BACKGROUND AND
AFFECT THEM A LOT, ACTUALLY, SO
THAT'S WHY I THINK -- THAT'S WHY
I MENTIONED COMPARE BEFORE AND
AFTER IS OUR MOST ROBUST
RESULTS.
>> HI, SO, DID I UNDERSTAND
CORRECTLY WHEN YOU SAID THAT BY
FUNCTIONAL ULTRASOUND, YOU FIND
INCREASED PREFRONTAL ACTIVITY IN
THE SHANK MUTANT WHEREAS ALL THE
OLD LIKE PHYSIOLOGY SLICES, AND
SO FORTH TOLD YOU SHOULD BE
EXPECTING THE OPPOSITE?
>> YEAH, SO, YES OR NO.
SO IN THE RESTING STATE AND IN
SLICE, YOU FIND REDUCE THE
SYNAPTIC FUNCTIONAL
CONNECTIVITY.
IN SLICE YOU FIND REDUCED
CORTICALE STRIATAL, BUT YEAH,
THAT'S TRUE, BUT IN FUNCTIONAL
ULTRASOUND, IF YOU DO RESTING
STATE, YOU ALSO FIND IT REDUCED,
IT'S ONLY BEHAVIOR TRIGGERED
ACTIVITY IS DIFFERENT.
>> I SEE AND YOU COULDN'T ASK
ABOUT THE BEHAVIORED TRIGGER TD
ACTIVITY, BECAUSE THE QUESTION
OBVIOUSLY IS HOW THIS AFFECTS
HOW WE THINK ABOUT A HUGE AMOUNT
OF EXISTING DATA THAT IS SORT OF
UNDERLYING YOUR WHOLE PICTURE OF
THE DISEASE ETIOLOGY, RIGHT?
>> WE ALSO DID 2-PHOTON IMAGING
IN MICE, IF YOU USE WHISKER
TRIGGERING THEY ARE ALSO HYPER
ACTIVE, IF YOU ANONSPECIFIC SIDE
EFFECTSITIZE MICE, THE PICTURE
MAY BE DIFFERENT.
>> OKAY.
THANK YOU.
>> OKAY 1 QUESTION FROM ONLINE,
THIS IS FROM SUE YING LEE, CAN
YOU TELL US GENE THERAPY 3
MONTHS VERSUS LATER, DO YOU SEE
DIFFERENCES IN RECOVERY?
>> THAT'S VERY GOOD QUESTION.
SO 3 MONTHS SINCE WE STARTED
VERY YOUNG IT'S A QUESTION, A
LITTLE DIFFICULT TO ANSWER
BECAUSE THE MARMOSET BEHAVIOR
CHANGES WHEN THEY GROW, SO IT'S
HARD TO SAY WHETHER IMPROVEMENT
IS 9 MONTHS IS BETTER THAN 3
MONTHS BECAUSE WE CANNOT MEASURE
EXACTLY SAME LEVEL, LIKE 3
MONTHS THEIR SOCIAL LEVEL MAY BE
DIFFERENT FROM 9 MONTHS, MAYBE
EVEN WILD-TYPE SO IT'S HARD TO
SAY SO WE DON'T HAVE ENOUGH DATA
TO ANSWER THAT QUESTION, BUT AT
3 MONTHS AFTER GENE THERAPY WE
ALREADY SEE VERY SIGNIFICANTLY,
SO AT THE END WE PUT A 3, 6, 9
MONTHS TESTING ALTOGETHER SO WE
DON'T SEE CLEAR SEPARATION, WE
ALSO DON'T SEE CLEAR SEPARATION
MALE AND FEMALE, WE ALSO DON'T
SEE CLEAR SEPARATION BECAUSE OF
THE SMALL NUMBER WHETHER 3
MONTHS INJECTION OR 9 MONTHS
INJECTION OR BETTER BECAUSE YOU
CAN COMPARE WHEN IT'S 12 MONTHS
WE DID NOT SEE, FOR THE
FUNCTIONAL MRI IT WAS WAY OVER 1
YEAR SO A LONG LASTING EFFECT.
>> OKAY, 1 THING I'M SUPPOSED TO
REPEAT THE CME CODE, SORRY FOR
THE INTERRUPTION, IT'S 57683,
THAT'S 57683.
SO YEAH, WE HAVE 1 MORE QUESTION
AND THIS QUESTION, I THINK A LOT
OF US HAVE THIS QUESTION, WHAT
IS THE THRESHOLD MIN NUM NUMBER
OF NEURONS TRANSDUCED BY THE AAV
VECTOR NEEDED FOR EFFECTIVE
RESCUE.
>> THAT'S A VERY GOOD QUESTION.
>> AND ALSO LIKE THE CELL TYPES.
>> YEAH, SO THAT'S A VERY GOOD
QUESTION, VERY TOUGH TO ANSWER,
WE -- MANY OTHER VS DONE
DIFFERENT STUDIES, WE DID OTHER
STUDIES, I THINK MY FEELING IS
IF THIS IS 15-20%.
I THINK IF YOU HAVE 5% OF
NEURONS, BRAIN WIDE, YOU WILL
SEE A SIGNIFICANT DIFFERENCE AND
10% DEFINITELY, WE HAVE LOCAL
INJECTIONS OF 10%, WE CAN SEE
VERY SIGNIFICANT CHANGE AND SO,
IT DEPENDS ON THE GENES.
FOR THE SHANK 3 FOR YOUNG
NEURONS ARE VERY PLASTIC, THEY
KNOW, THEY WILL SHIFT THEIR
WEIGHT TO THE FUNCTIONAL NEURONS
INSTEAD OF AVERAGE, SO THIS WAS
WITH THE SYNAPSE CAN WORK REALLY
WELL, SO 5-10% WILL PROBABLY
MAKE A DIFFERENCE?
YOUR OTHER QUESTION?
I FORGOT.
>> OKAY, SO HAVE YOU LOOKED AT
SLEEP DISORREREDDERS AND THEIR
RESCUE IN TRANSGENIC --
>> NEURON TYPE, SHANK 3 EXPRESS
IN EXCITED NEURONS AND INHIBITED
NEURONS BECAUSE THEY BOTH HAVE
EXCITE O SYNAPSE AND THESE
SYNAPSE BOTH NEED SO WE USE THE
SYNAPSE SIN PROMOTER SO IT GOES
TO BOTH TYPES AND THEY MATCH,
THE PERCENTAGE MATCHED
ENDOGENOUS AND WE TEST THIS.
SORRY.
>> OH, HI, WOBDERFUL TALK,
REALLY SHOWS THE VALUE OF
CONSTRUCT VALIDITY, YOU KNOW AND
YOU TALKED ABOUT SOME OF THE
RARE MUTATIONS THAT ARE
ASSOCIATED WITH SCHIZOPHRENIA,
AND I'M WONDERING IF YOU GUYS
ARE THINKING ABOUT WAYS TO MODEL
COPY NUMBER VARIANCE.
NYEAH, SO VERY GOOD QUESTION.
SO WHEN JOSH GARDENER WAS HERE
HE WAS VERY INTERESTED IN THE 22
Q DELETION, YOU CAN CAN MODEL
THEM.
WE CAN DO LARGE DELETION NO
PROBLEM, DUPLICATION IS MORE
DIFFICULT SO WE TESTED THAT FOR
I THINK 6 KB, 6 MEGABASE,
SMALLER EDITION, IT WORKS WELL
IN EMBRYOS BUT WE END UP WITH
NOT REACHING THE MARMOSET MODEL
BECAUSE IT ALSO HAVE CARDIAC
ISSUE.
AND WE'RE WORRIED THAT THE
MONKEY WILL NOT SURVIVE SO WE
DID NOT.
SO TECHNOLOGICALLY, YES WE CAN
DO CMEs.
>> OKAY, SO 1 ONLINE QUESTION
ABOUT SLEEP DISSEREDDER
PHENOTYPES.
>> YEAH, OH, THAT'S -- SO WE DID
A TEST AND WE DID NOT SEE
BECAUSE WE HAVE THESE VIDEOED WE
DID NOT SEE ANY SIGNIFICANT
DEFECT IN SLEEP, WE DON'T KNOW
WHETHER IT'S MONKEYS ARE
DIFFERENT FROM HUMAN, WE DON'T
KNOW IF IT'S IN THE HOUSING
CONDITION THEY DON'T HAVE
NATURALISTIC SLEEP, HAVE YOU
TURN ON AND TURN OFF, SO WE
DON'T KNOW WHICH IS THE CAUSE,
WHETHER IT'S BECAUSE OF HUMAN
MONKEY DIFFERENCE OR WHETHER
IT'S BECAUSE THE HOUSING
CONDITION DIFFERENCE, WE DON'T
KNOW ABOUT YOU WE DID TEST IT,
WE DID NOTICE SLEEP DIFFERENCE.
LAST QUESTION FROM YU GU, ARE
ALL SHANK 3 MUTATIONS FOR AUTISM
LOSS OF FUNCTION?
>> SO FAR, YES, MOST OF THEM ON
LE LESIONS ACTUALLY.
YEAH.
>> OKAY, THAT'S IT.
THANK YOU EVERYONE FOR YOUR TIME
AND COMING.
[ APPLAUSE ]