>> WELCOME TO THE WEDNESDAY
AFTERNOON LECTURE SERIES, I AM
MICHELLE ANTWON, CHIEF OF THE
SECTION ON NEUROCIRCUITS AT NIH.
IT IS MY GREAT PLEASURE TO
INTRODUCE TODAY'S DISTINGUISHED
SPEAKER DR. RICHARD TSIEN,
DR. TSIEN DESCRIBES HIMSELF AS A
CARDIAC AND NEURONAL
FUNCTIONALIST AND WE ALSO SEE
HIM AS A PIONEER IN THE VOLTAGE
CALCIUM CHANNELS STUDIES.
HIS INFOIVATIVE WORK DELVES INTO
THE MECHANISMS AND ROLES OF
THESE VESSELS IN SIN APTICAL
FUSION, CELL SIGNALING PATHWAYS
AND YOUR OVERARCHING IMPACT ON
NEUROTRANSMITTER RELEASE,
SYNAPTIC TRANSMISSION AND
SYNAPTIC PLASTICITY IN BOTH THE
HEALTHY AND DISEASED BRAIN.
FOR THOSE WATCHING REMOTELY, WE
ENCOURAGE YOU TO PARTICIPATE IN
THE QUESTION AND ANSWER SESSION
AFTER 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
YOUR NAME AND QUESTION AND WE
WILL RELAY THIS MESSAGE TO
DR. STIEN AT THE END OF HIS
LECTURE.
YOU CAN SUBMIT A QUESTION ANY
TIME AND I HOPE YOU DO SO.
FOR THOSE OF YOU IN THE
LIPSETT AMPHITHEATRE TODAY, YOU
CAN USE THE MICs ON THE LEFT
AND RIGHT.
WE ARE ALSO OFFERING CONTINUING
MEDICAL EDUCATION CREDITS FOR
THIS PRESENTATION, THE CME CODE
FOR TODAY IS 50095.
THAT'S 50095.
SO LET'S RESUME, OUR ESTEEMED
SPEAKER TODAY IS DR. RICHARD
TSIEN, WHO IS THE MILLER
PROFESSOR OF NEUROSCIENCE OF
CHAIR OF DEPARTMENT OF
NEUROPHYSIOLOGY AND
NEUROSCIENCE.
HE EARNED HIS BACHELOR OF
SCIENCE AND MASTER'S OF SCIENCE
DEGREE IN ELECTRICAL ENGINEERING
FROM THE MASSACHUSETTS INSTITUTE
OF TECHNOLOGY.
SUBSEQUENTLY, HE WAS AWARDED A
ROAD SKOALARSHIP TO PURSUE HIS
DR. OF PHILOSOPHY DEGREE, IN
BIOPHYSICS AT THE UNIVERSITY OF
OXFORD.
WHERE HE CONDUCTED RESEARCH
UNDER THE GUIDANCE OF PROFESSORS
DENNIS NOBLE AND JULIAN JOCK,
AFTER OXFORD DR. TSIEN JOINED
YALE UNIVERSITY SCHOOL OF
MEDICINE AS AN ASSISTANT
PROFESSOR IN THE DEPARTMENT OF
PHYSIOLOGY.
EVENTUALLY OBTAINING THE RANK OF
FULL PROCESSOR WITHIN 9 YEARS.
AFTER SPENDING 18 YEARS AT YALE,
HE RELOCATED TO STANFORD
UNIVERSITY IN 1988 WHERE HE
ESTABLISHED THE DEPARTMENT OF
MOLECULAR AND CELLULAR FIELD
FUNCTIONS YOJ AND SERVED AS ITS
INAUGURAL CHAIRMAN, HERE HE ALSO
SERVED AS A CO DIRECTOR FOR THE
STANFORD BRAIN RESEARCH CENTER.
IN 2011, DR. TSIEN LEFT STANFORD
AS AN EMERITUS FACULTY MEMBER
AND TRANSITIONED TO THE NEW YORK
UNIVERSITY [INDISCERNIBLE]
MEDICAL PROFESSOR ASSUMING THE
POSITION OF THE NEUROLOGICAL
DIRECTOR OF THE SCIENCE
EN--STRATEGIESITUTE.
IN TBENT 15 HE WAS NAMED
INAUGURAL SCIENTIFIC DIRECTOR OF
THE [INDISCERNIBLE] INSTITUTE
FOR PARKINSONS AND MOVEMENT
DISORDER AT NYU.
DR. TSIEN HAS FEARLESSLY FORGED
A TRAIL OF GROUPED BREAKING
RESEARCH.
HIS RESEARCH DOCUMENTED IN OVER
300 PEER REVIEWED JOURNAL
ARTICLES HAS SIGNIFICANTLY
ADVANCED OUR UNDERSTANDING OF
HOW NEUROTRANSMITTERS, DRUGS OF
ABUSE AND MOLECULAR ALTERATIONS
REGULATE CALCIUM CHANNELS.
DR. TSIEN'S CONTRIBUTIONS EXTEND
TO ELUCIDATING THE MECHANISMS
THAT FACILITATE ADAPTATION OF
SYNAPTIC STRENGTH OR ALTERATIONS
IN NEURONAL GENE EXPRESSION,
THROUGH CHANGES IN SYNAPTIC
TRANSMISSION, AND
DEPOLARIZATION, THE PRIME BRAIN
NETWORKS FOR LEARNING AND MEMORY
FUNCTIONS.
ADDITIONALLY HIS WORK HAS
YIELDED VALUABLE INSIGHTS INTO
THE MECHIMISMS THAT,A LOW US TO
ADAPT TO SUDDEN OR PROLONGED
CHAIMPLES IN ACTIVITY LEVELS.
THE IMPLICATIONS OF HIS RESEARCH
REVERBERATE ACROSS DIVERSE
CLINICAL DOMAINS, ENCOMPASSING
DISEASES SUCH AS ALZHEIMER'S
DISEASE, AUTISM, NEUROLOGICAL
DISORDY SAYS LIKE MIGRAINE,
ATTACKSIA, DYSTONIA, MORE OVER
HIS FINDING ARE PERINENT TO
TERMINAL CONDITIONS SUCH AS
SUDDEN UNEXPECTED DEATH IN
CHILDHOOD, DR. TSIEN IS ALL THE
CO-AUTHOR FOR THE SEMINOLE
TEXTBOOK, ELECTRIC CONFLUENCE IN
EXCITABLE CELLS, A FUNDAMENTAL
RESOURCE FOR STUDENTS STUDYING
BIOPHYSICS, HIS LIST OF
ACCOLADES INCLUDES THE ROLE
SCHOLARSHIP TO THE INVESTIGATOR
AWARD, THE KAISER AWARD, THE
PRIZE, THE CART WRIGHT PRIZE,
THE AXLE ROD PRICE, THE JAMILA
ROD PRICE, THE DIRECTORSHIP,
DISTINGUISHED JOHN BURDEN PRICE
LECTURE OF THE UNIVERSITY OF
OXFORD.
NOTABLY HE HAS BEEN ELECTED AS A
MEMBER OF THE NATIONAL ACADEMY
OF MEDICINE, THE NATIONAL
ACADEMY OF SCIENCES, THE
INSTITUTE OF MEDICINE, THE
AMERICAN ACADEMY OF ARTS AND
SCIENCES AND THE ACADEMIA
SENICA, WHICH IS THE NATIONAL
ACADEMY OF THE REPUBLIC OF
CHINA.
THE TITLE OF DR. TSIEN'S TALK
TODAY IS LEVERAGING GENETICS AND
CELL SIGNALING TO DECIPHER
DISORDERS OF EXCITABILITY.
PLEASE WELCOME DR. TSIEN.
[ APPLAUSE ]
NAND AS DR. TSIEN IS APPROACHING
THE PODIUM, WE HAD A MIX UP WITH
THE LIVE FEEDBACK FORM, SO FOR
THOSE WATCHING ONLINE, IF YOU
DON'T SEE IT MAYBE IN A FEW
MINUTES, RESILIENCE LOAD YOUR
VIDEOCAST SYSTEM AND THE BUTTON
WILL BE SHOWING UP IN A FEW
MINUTES AND THE CME CODE IN CASE
YOU MISHEARD WAS 50110.
50110.
THANK YOU.
ALL YOURS DR. TSIEN.
>> THANK YOU VERY MUCH,
DR. ANTWON, AND THANK YOU
DR. SHORE AND THANKS TO DIANE
GOMEZ FOR ARRANGING THIS VISIT.
IT'S A BIG PLEASURE FOR ME TO BE
HERE AT NIH AND SEE FOLKS WELL
RECOVERED FROM THE PANDEMIC AND
I MET WONDERFUL PEOPLE TODAY,
TRAINEES AND FACULTY AND I HOPE
TO MEET SOME MORE DURING THE
REST OF THE DAY.
I WOULD LIKE TO TALK TO YOU
ABOUT A TOPIC THAT'S VERY
RELEVANT TO NIH AND THAT IS HOW
WE'RE GOING TO TAKE THE
DISCOVERIES FROM GENETICS AND
USE THEM TO UNDERSTAND CELL
SIGNALING AND THEREFORE TO
DECIPHER DISORDERS OF
EXCITABILITY, INCLUDING
EPILEPSY, AND AUTISM AND
EVENTUALLY SCHIZOPHRENIA.
I HAVE NO DISCLOSURES TO MAKE
NOR DO MEMBERS OF MY LAB, AND WE
HAVE SUPPORT FROM A NUMBER OF
ORGANIZATIONS MYSELFATIONS
INCLUDING BRAIN INITIATIVE,
NINDS, NIMH, NIDA, NIGMS, BUT
NOT SPECIFICALLY FOR THE WORK ON
CBD THEY WILL TELL YOU ABOUT, A
NUMBER OF DIFFERENT FOUNDATIONS
HAVE BEEN VERY GENEROUS TO US.
IN TODAY'S LECTURE I WOULD LIKE
TO SHOW OW GENETIC INSIGHTS
PROMPT NEUROLOGICAL APPROACHES
TO FUNCTION IN PATHOPHYSIOLOGY.
YOU CAN READ THIS, I GUESS.
DELINEATE WHICH EXCITATORY
SIGNALING MOLECULES COORDINATE
EXCITATORY AND HIPAAATORY
FUNCTION.
TALK A LITTLE BIT ABOUT SUDDEN
AND UNEXPECTED DEATH IN
CHILDHOOD AND EVENTUALLY MAKE
PROMISSORY NOTES ABOUT HOW WE
WILL TAKE WHAT WE KNOW ABOUT
SIGNALING PROTEINS, AND APPLY IT
TO A BIG DISEASE LIKE
SCHIZOPHRENIA.
NOW, MANY PEOPLE KNOW THAT
NEUROLOGICAL AND PSYCHIATRIC
DISORDERS SHARE GENES AND MAY
EVEN SHARE THERAPEUTIC
POTENTIAL.
IN THIS SLIDE FROM JOE BUXBAUM'S
WORK, YOU CAN SEE A DIAGRAM
SCHEMATICALLY DEPICTING HOW THE
DIFFERENT DISEASES LIKE
SCHIZOPHRENIA, INTELLECTUAL
DISABILITY AND AUTISM, ACTUALLY
OVERLAP WITH EPILEPSY IN THEIR
GENETIC BASIS.
MARCUS I'M HAVING TROUBLE
FINDING THE POINTER.
WAIT A SECOND.
HERE I GO.
CAN YOU SEE THE POINTER?
OKAY, AND INDEED, BILL
[INDISCERNIBLE]'S LAB CAME UP
WITH THE IDEA THAT AS 1 EXAMPLE
OF THIS KIND OF OVERLAP, A
MEDICINE THAT WAS PREVIOUSLY
USED FOR EPILEPSY OR CBD, OR
KAN--KANA A--BITS KIOLE WAS
EFFECTIVE IN TREATING A LEVEL OF
AUTISM IN MOUSE LEVEL.
THIS IS THE MOUSE NAV1.1, WHO'S
NEURONS ARE NOT BEHAVING
PROPERLY AND THIS EXPERIMENT BY
NEFFA STELLA AND COLLEAGUES IT
SHOWS IF YOU USE CBD AT
CLINICALLY EFFECTIVE DOSES IN
MEN, CAN YOU SEE IMPROVEMENT IN
THE SOCIAL BEHAVIOR OF A MOUSE
BRENGS IT UP TO SORT OF CONTROL
LEVELS AS YOU INCREASE THE DOSE.
SO THE POSSIBILITY IS THAT AN
ANTISEIZURE MEDICATION MIGHT
CROSS OVER AND AFFECT A SOCIAL
DEFICIT THAT WOULD TRADITIONALLY
BE THE PROVINCE OF
PSYCHIATRISTS.
NOW I WOULD LIKE TO TALK ABOUT
CBD BECAUSE IT RINGS BELLS WITH
ENDOKACCT NABBOIDS AND IT IS A
MEDICINE THAT'S BEEN FDA
APPROVED.
ON 1 HAND WE HAVE THE CONTRAST
BETWEEN THE PSYCHOACTIVE
INGREDIENT OF CANNABIS, KELTA 9
THH AND KAN--KANAA BIN OIL, THE
NONPSYCHOTROAPIC, AND THAT'S
IMPORTANT THING FOR ITS USE AS A
MEDICINE.
THC AS A NUMBER OF PEOPLE ARE
STUDYING, CAN HAVE A NUMBER OF
DIFFERENT EFFECTS, INCLUDING
INCREASING HUNGER WHEREAS ON THE
OTHER HAND CANNABITIOL CAN
COUNTERACT THESE SIDE EFFECTS OF
DELTA 9 AND FINALLY THE CB1
RECEPTOR IS THE TARGET OF
INVESTIGATION OF MANY PEOPLE AT
NIH INCLUDING GEORGE KUNOS, BUT
THE CBD HAS VERY LOW AFFINITY AT
THE CB1 RECEPTOR.
SO THESE CHEMICALS ARE VERY
CLOSE TO EACH OTHER, THE KEY
DIFFERENCE LIES IN THIS OXYGEN
SPANNING LINK BETWEEN THESE 2
WHICH EXISTS IN CBD.
NOW THE MEDICAL MOTIVATIONS ARE
ABUBD ANT.
FDA AS PROVED FOLLOWING A SERIES
OF DOUBLE BRINEDDED CLINICAL
STUDIES FOR ITS USE IN THE
TREATMENT OF DRAVET SYNDROME
WHICH IS A FORM OF INTRACTABLE
EPILEPSY AND CBD CAUSED A 40%
REDUCTION IN SEIZURE FREQUENCY,
LIKEWISE IN LENOIR OX
[INDISCERNIBLE] SYNDROME, A
ANOTHER FORM OF EPILEPSY, A
SIMILAR MODIFICATION WAS
CHAIMPLED.
SO IT'S ALREADY BEEN APPROVED
AND ON THE STREET MANY PEOPLE
ARE USING IT FOR ANYTHING FROM
ANXIETY TO INSOMNIA AND NOT
WORRYING ABOUT WHAT DOSE THEY'RE
USING OR HOW GOOD THE DRUG THERE
IS, SO THERE'S ACTUALLY AN
EXPERIMENT GOING ON AS WE OUT ON
THE STREET OF PEOPLE
EXPERIMENTING WITH THIS AGENT
AND BECAUSE IT DOESN'T PRODUCE A
HIGH AND IS NOT ILLEGAL, THERE'S
A LOT OF TESTING GOING ON.
THE QUESTION IS, HOW DOES IT
ACTUALLY WORK AND IF WE LEARN
HOW IT REALLY WORKS CAN WE APPLY
IT MORE INTELLIGENTLY AND
TARGETED WAY TO AFFECT DISEASE
OF THE BRAIN.
THIS IS WORK DONE BY WHAT I
MIGHT CALL THE CBD TEAM HEADED
BY EVAN ROSENBERG, AN MD-Ph.D.
STUDENT DOING CLINICAL TRAINING
AT PENN, AND SIMON CHAMBER LAND,
AN ELECTROPHYSIOLOGIST
EXTRAORDINARY ALONG WITH ERICA
NEBET, WHOSE GONE ON TO MEDICAL
SCHOOL, ALEJANDRO SALAH, AND
GOUOLIING TIAN AND XIAOHAN WANG,
AND I WILL TALK ABOUT THEM SOON.
WE WERE JOINED BY HELEN SHARP
MAN, [INDISCERNIBLE] JANE AND
YURI, [INDISCERNIBLE], AND DREW
JONES AND REBECCA ROSE AND WE
TEAMED UP WITH A GROUP AT THE
UNIVERSITY OF REDDING WHOSE WORK
I WILL SHOUT OUT ABOUT ALONG
WITH OTHER PEOPLE IN THE UK
TOGETHER WITH GW
PHARMACEUTICALS.
SIMON IS GOING ON TO A FACULTY
JOB AT THE UNIVERSITY OF
PITTSBURGH.
NOW THERE ARE MULTIPLE
CANDIDATES FOR CBD ACTION EMPLOY
INCLUDING THE CB1 RECEPTOR, OR
ENZYMES THAT ARE INVOLVED IN THE
PROSECTION OF AEA, IT CAN ALSO
HAVE ACTIONS ON OTHER LIGANDATED
TRANLES, TRIP KAN--KANA CHEALS,
THELP TRIP CALCIUM CHANNELS BUT
TODAY WE WILL FOCUS ON ITS
ACTION IN 1 PARTICULAR TARGET IN
ORPHAN G-PROTEIN RECEPTOR CALLED
GPR55.
THIS MOLECULE LIVES IN THE
MEMBRANE AND RESPONDS TO
STIMULATION BY ITS ENDOGENOUS
LIAISON IMAND, CALLED LPI, THAT
AGANISM IS BLOCKED BY CBD AS A
COMPETITIVE ANTAGONIST.
SO THIS COMBINATION OF A NATURAL
LIGAND, RECEPTOR AND ANTAGONIST,
IS THE BASIS FOR THIS STORY,IME
GOING TO TELL YOU IN THE FIRST
PART OF MY TALK.
WHY ARE WE INTERESTED IN THESE
AGENTS?
BECAUSE OF GENETICS.
GENETICS TELLS US FIRST OF ALL
THAT LPA IS INTERESTING, THAT'S
BECAUSEIOSEVERE MITRAL GLEASON
TEAMED UP WITH GROUPS IN
PAKISTAN AND OTHER COUNTRIES IN
ASIA AND STUDIED THE METABOLISM
OF LPI.
LPI IS PRODUCED AS A PRODUCT OF
FOSTER NURSED FOCUSED ON TITLE
INOCITOLL, A LIPID PRESENT IN
ALL MEMBRANES AND VERY IMPORTANT
AS A REPOSITORY FOR
[INDISCERNIBLE] ACID.
WHEN CLEAVAGE IS DONE FOR FOSTER
NURSED POETIC LIPASE 2, THE
MODEL CITIZENETY IS CLIPPED OFF
LEAVING YOU WITH AN ACEL CARBON
CHAIN LEADING TO LPI, AND THE
SIN KRETTIC ENZYME FOR THIS IS
CALCIUM DEPEND EPT.
SO AS A RESULT OF ELECTRICAL
ACTIVITY IN NEURONS, THERE IS
MODEL CITIZEN DUKS OF LPI.
THE REVERSE ENZYME CALLED
LPUNISHING I ACE ILLEGALS
TRANSFER ACE, AND IT'S THE ONLY
ENZYME KNOWN TO CONVERT LPI FROM
THIS ACTIVE FORM BACK BO THE PI
RESERVOIR.
WHEN YOU HAVE MUTATIONS IN THIS
ENZYME, AND THAT WAS STUDIED IN
HIGHLY INBRED FAMILIES IN
VARIOUS COUNTRIES, COUSINS
MARRYING EACH OTHER JUST LIKE IN
THE ROYALTY OF THE EUROPEAN
CAPITALS, THE RESULT IS
ENACTIVATING MUTATIONS THAT ARE
DOUBLED UP ON BOTH ALLELES AND
THEREFORE, CAUSE EPILEPSY,
AUTISM, AND INCOLLECTUAL
DISABILITY, SO THERE WAS GENETIC
EVIDENCE THAT THE ABSENCE OF
BREAK DOWN OR DISPOSAL OF LPI
COULD CAUSE DISEASE, HOWEVER,
THE PEOPLE WHO DID THE STUDY
INTERPRETED, NOT IN TERMS OF THE
BAD THINGS THAT TOO MUCH LPI
WOULD DO, BUT RATHER THE BAD
THINGS THAT MIGHT HAPPEN IF YOU
HAD TOO LITTLE PI, AND WE'RE
GOING TO MAKE THE CASE THAT IT'S
ACTUALLY LPI THAT IS THE BAD
ACTOR AND THAT LPI ITS KNOWN
RECEPTOR GPR55 AND THAT EFFECT
IS ANTAGONIZED BY CBD, SO
GENETICS LEADS US TO THE LIGAND
AND ASSOCIATION THROUGH WORK OF
OTHER PEOPLE TELLS US ABOUT GPR
55 AND C BD.
SO IF YOU USE A KNOCK OUT MOUSE,
KINDLY PROVIDED TO US BY KENNETH
MACKEY OF INDIANA UNIVERSITY,
YOU CAN SEE THAT GPR 55 IS
ABSENT IN A REGION OF BRAIN,
IMPORTANT FOR EPILEPSY AND
LEARNING AND MEMORY CALLED A
HIPPOCAMPUS.
SO HERE WE HAVE STAINING FOR GPR
55, HERE WE HAVE ONLY STAINING
FOR THE NUCLEUS OF THE CELL AND
NO GPR 55 AND IF YOU TAKE MICE,
AND TREAT THEM WITH PENTINE
REGIMEN O LEAN [INDISCERNIBLE]
TO INDUCE SEIZURES, YOU FIND
THERE'S A DOSE REDUNDANT
INDUCTION IN SEIZURES IN
RESPONSE TO INCREASING AMOUNTS
OF CBD BUT THAT DOZE DEPENDENCE
IS GONE IN THE GPR55 KNOCK OUT
EMPLOY SUGGIEST THAGOREAN GPR55
MIGHT HAVE SOMETHING TO DO WITH
THE SEIZURES.
WHAT HAPPENS TO NEURONAL
FUNCTION?
WE STUDIED THAT IN SLICES FROM
THE HIPPOCAMPUS THAT HAVE THE
CLASSICAL REGIONS SCRIBED BY
RAMONEIC [INDISCERNIBLE] ET ALL,
WHICH IS THE TARGET OF SYNAPSES
GIVEN OFF BY CA3 PARAMETAL ISS
THAT SEND AXONS ALONG THE
SHAFTER COLLATERALS TO
INNERIVATE THE CAC1 NEURONS AND
WHEN WE STIMULATE THOSE AXONS
WITH AN EXTRA CELLULAR STIMULUS,
WE FIND THAT THE STIMULUS IS
FOLLOWED BOY ACTION POTENTIALS
THAT RECORDED AS EXTRA CELLULAR
CURRENTS SHOWN HERE AND THAT LPI
INCREASES THE INCIDENCE OF
SUCCESSFUL FIRING BY ABOUT 50%
AND AN EFFECT THAT DISAPPEARS IF
YOU MAINTAIN EXPOSEDDURE TO LPI.
SO WE CAN THEN LOOK AT THE
EFFECT OF CBD AND SHOW THAT THE
EFFECT OF THE KNOCK OUT IS
MIMICKED BY THE EFFECTIVE CBD.
SO THIS IS EXACTLY WHAT YOU
WOULD EXPECT IF A RECEPTOR,
G-PROTEIN RECEPTOR AND CBD WAS
THE ANTAGONIST FOR IT SO WHEN
YOU HAVE CBD PRESENT, IT'S AS IF
THERE WAS SO RECEPTOR AT ALL AND
LPI WAS THE ENDOGENOUS LIGAND
THAT INCREASE THE RATE OF ACTION
POTENTIALS BY ABOUT 50%.
THIS IS HARDLY EPILEPSY BUT IT'S
DONE UNDER MILD CONDITIONS WHERE
WE'RE LOOKING TO SEE THAT
THERE'S A PHYSIOLOGICAL EFFECT.
NOW TO UNDERSTAND HOW CBD HAD
THIS ACTION ON TRANSMISSION,
FROM THE SHAFT COLLATERALS TO
THE CAC1 NEURONS, WE RECORD
QUANTAL SYNAPTIC EEIVETS FROM
THE CAC1 NEURONS THESE ARE CALL
MINIATURES OR MEPSCs, THIS IS
TRIGGERED BY GLUTEA MITTURGIC
TRANSMISSION.
AND IF WE SHRINK THESE DOWN AND
CHANGE THE TIME SCALE TO RECORD
OVER A MINUTE RATHER THAN 2
MILLI SECONDS, WE SEE
SPONTANEOUS RELEASE OF QUAWPT
UMKC FROM THE PRESYNAPTIC
TERMINALS AND THAT LPI GREATLY
INCREASES THE FREQUENCY OF THESE
QUANTA AND THE EFFECT OF THIS IS
REVERSIBLE AND THIS IS THE TIME
PLOT SHOWING THAT ON AVERAGE,
THERE'S SOMETHING LIKE A 70%
INCREASE IN FREQUENCY WHICH IS
TRANSIENT, AND NO CHANGE IN THE
AMPLITUDE OF THE SYNAPTIC
EVENTS.
EVIDENTLY THE PRESYNAPTIC
TERMINAL IS RELEASING MORE
QUANTUM PER UNIT PERIOD OF TIME
AND THIS LEADS TO A TRANSIENT
INCREASE IN EXCITATORY
TRANSMISSION.
THIS IS COMPLETELY BLOCKED BY
CBD, SO FITTING WITH THE IDEA
THAT IT'S PLOKING THE RECEPTOR
FOR THESE DOWN STREAM EVENTS
PRESUMABLYOT PRESYNAPTIC
TERMINAL.
PREVIOUS WORK ON THIS BY DIMITRI
[INDISCERNIBLE]'S LAB WAS
CONFIRMED BY THIS SLIDE, THIS
FIGURE.
WE WENT ON TO LOCK AT CHANGES IN
THE INHIBITORY TRANSMISSION,
AFTERALL THIS IS ONLY A
TRANSIENT CHANGE.
PERHAPS THERE WAS A CHANGE IN
INHIBITORY TRANSMISSION THAT WAS
MORE SUSTAINED, SO WE RECORDED
INHIBITORY EVENTS LIKE THIS,
ONLY NOT CARRIED BY SODIUM IONS
BUT CARRIED BY CHLORON IONS AND
WE FOUND IN THIS CASE, THE
FREQUENCY OF EVENTS WAS NOT VERY
MUCH CHANGED.
WHAT WAS MAINLY CHANGED WAS THE
AMPLITUDE OF THE EVENTS, YOU CAN
SEE ALL THESE EVENTS ARE SMALLER
AND THAT SHOWN HERE ON A SLOWER
TIME SCALE OVER 10-20 MINUTES,
THERE'S PROGRESSIVE DEVELOPMENT
IN THE EFFECT.
THERE'S A SLIGHT DECREASE IN
FREQUENCY AND THERE'S A VERY
CLEAR DECREASE IN AMPLITUDE OF
THE SYNAPTIC EEIVETS AND THIS IS
ALSO BLOCKED BY LPI, SO
EVIDENTLY THE RECEPTOR IS
COUPLED TO CHANGES BOTH IN
EXCITATORY AND INHIBITORY
TRANSMISSION, AND IN SLIDES I
DECIDED TO LEAVE OUT, BECAUSE
IT'S IN OUR PAPER, AND TAKES
ABOUT 5 MINUTES TO EXPLAIN, WE
WORKED OUT THE MECHANISM OF
THIS, WE FOUND THAT THE NUMBER
OF GABBA RECEPTORS IN THE POST
SYNAPTIC CELL, IN THE CAC1
PARAMETAL IS WAS DECREASED AND
THAT THIS INVOLVED A PERFECTLY
CONVENTIONAL MODULATION OF
INHIBITORY TRANSMISSION.
SO EVAN THEN WENT ON TO STUDY
EVOKE STIMULATION BY STUDYING
THE SAME INPUT THAT LED TO THE
EVOKED ACTION POTENTIALS,
RECORDING FROM THE PARAMETRAL
NEURON BUT NOW IN RESPONSE TO AN
ELECTRICAL INPUT AND THIS SHOWED
RESULTS COMPATIBLE WITH THESE,
AND THAT PRIMARILY THAT WAS
CAUSED AN INCREASE IN THE
EVOCKED TRANSMISSION, AND IT WAS
MIMIC THE GPR55 RECEPTOR THERE
WERE ALSO CHANGES IN PAIR POST
MODULATION, WHICH ARE HERE FOR
THE OFFICKIAN ADOS WHO ARE
INTERESTED IN FIGURING OUT
WHETHER THIS IS PREOR POST
SYNAPTIC, NOT ONLY DID WE SEE
CHANGES IN EXCITATORY
TRANSMISSION BUT WE'RE PLEASED
TO SEE A CHANGE IN THE
INHIBITION WHICH IS PRODUCED
WHEN THE COLLATERAL GIVE OFF
BRANCHES IN INNERIVATE
INHIBITORY NEURONS THAT INTURN
SYNAPSE ON TO THE PARAMETAL ISS,
THIS IS CALLED DISCIPLINARY
SYNAPTIC INHIBITION, AND WE SAW
A BIG DECREASE IN THE SIGNIFY
SYNAPTIC INHIBITION IN RESPONSE
TO CBD, SO THIS IS THE DECREASE,
IT'S SOMETHING LIKE A 60 OR 70%
DECREASE.
AND ONCE AGAIN, THIS IS
PREVENTED BY BLOCKING THE
RECEPTORS WITH CBD, IT'S
MIMICKED BY KNOCKING OUT THE
RECEPTOR AND THE CHANGES WE SEE
IN AMPLITUDE ARE PARALLEL BY
CHANGES IN PAIRED PULSE
MODULATION, THERE IS A BIG
INCREASE IN HOW MUCH BIGGER THE
SECOND RESPONSE IS TO THE FIRST,
INDICATING A RECOVERY OF RELEASE
PROBABILITY AND THAT ONCE AGAIN
IS BLOCKED BY CBD AND MIMICKED
BY THE KNOCK OUT.
SO EVIDENTLY, RELEASE
PROBABILITY IS GOING WAY DOWN IN
RESPONSE TO LPI.
O WE WENT THROUGH THIS, I WON'T
TAKE YOU THROUGH IT, IT'S
SHOWING THAT THE INTENSITY IN
CLUSTERS HAS GONE DOWN AND
THAT'S BECAUSE GEFFERIN HAS GONE
DOWN, AND THE GABBA 2 HAS GONE
DOWN I WILL SKIP THAT TO SAVE
TIME.
NOW THE KEY QUESTION IS HOW DOES
THIS PHENOMENA RELATE TO
EPILEPSY, IS THERE SOMETHING
AUTOCATALYTIC OR REGENERATIVE
ABOUT THIS.
WELL, WE WERE VERY GRATIFIED TO
FIND THAT IN RESPONSE TO A
SEIZURE THE CONCENTRATION OF
GPR55 RECEPTORS WAS GREATLY
INCREASED AND THAT AFFECT WAS
BLOCKED BY CBD, SO THIS IS NOT
JUST YOUR COMMON GARDEN VARIETY
RESPONSE WHERE THE RECEPTORS
REMAIN THE SAME, AND THE
SIGNALING THEY DO IS WHAT
CHAIMPLES.
HERE NOT ONLY IS THE SIGNALING
CHAIVENLGING BUT ALSO THE NUMBER
OF RECEPTORS THEMSELVES ARE
CHANGING IN A SEIZURE DEPENDENT
AND CBD DEPENDENT WAY THIS IS
THE GROUP DATA SHOWING THAT IN
CAC1 AND CA3, AND LESS
PROMINENTLY IN DENTAIMIEROUS,
THERE'S A CLEAR INCREASE IN THE
LEVEL OF RECEPTOR WHICH IS
REVERSED BY CBD.
SO THIS MEANS THERE'S AN
INHERENT LOOP BACK FEATURE
BECAUSE THE NUMBER THAT'S
RESPONDING IS INCREASED.
AND LIKEWISE WHEN WE DID
EXPERIMENTS USING RTPC R,
AGAINST THE MRNA THAT ENCODES
WITH THE SEIZURE, THIS ALSO
DECODES AND DECREASED AND EVEN
MORE REVERSED BY THE PRESENCE OF
CBD.
AS IF THERE WERE AN AMBIENT
LEVEL OF LPL PRESENT IN THE
SLICE, EVEN BEFORE CBD WAS
APPLIED OR AN AMBIENT LEVEL OF
THE RECEPTOR.
SOPHISTICATED HOW ABOUT THE
LIGAND, IS IT ALSO INFLUENCED BY
THE FACT THAT YOU HAVE A
SEIZURE?
WELL WE MADE AN ACUTE SEIZURE
WITH [INDISCERNIBLE], AND IT WAS
ELEVATED BY THE SEIZURE AND MORE
THAN REVERSED BY THE PRESENCE OF
CBD, THIS WAS DONE BY MASS SPEC
AGENTS AND OUR LIPID OMICS CORE
AND I WANT TO POINT OUT THAT THE
ISOFORM, PARTICULAR ISOFORM THAT
IS ASHINGED IS THE 180 ISOFORM,
18 CARBONS, 0 DOUBLE BONDS, THIS
IS THE CLASSIC ALLELE FORM OF
LPI THAT WAS THE VERY FORM THAT
WAS INCREASED IN RESPONSE TO THE
KNOCK OUT OF THE ENZYME THAT
RECONVERTS THIS BACK INTO PI.
SO SO THIS IS AN ALIGNMENT
BETWEEN GENETICS AND THE
BIOCHEMISTRY, THE BIOCHEMISTRY
IN THE SLICE SAYS THAT EPILEPTIC
SEIZURES REPRODUCE THE ACT THAT
IS PRODUCED BY THE RESULT IN THE
CONSANGUINEOUS FAMILIES AND I
FIND THIS VERY SATISFYING
BECAUSE IT PROVIDS SOME EVIDENCE
THAT IT'S LPI AND NOT PI THAT IS
THE KEY FACTOR UNDERLYING THE
MANIFESTATIONS OF AUTISM,
EPILEPSY AND INTELLECTUAL
DISABILITY IN THOSE FAMILIES.
SO TO SUMMARIZE WHAT I TOLD YOU
SO FAR.
LPI ACTS ON A RECEPTOR, THE
RECEPTOR ACTS AS THE INVITATION
AND SIMULTANEOUSLY SHUT DOWN
THE, LPI, IF YOU PRESS DOWN ON
THE ACCELERATOR AND LET GO OF
THE BREAK, YOUR CAR IS GOING TO
GO FASTER.
THAT PRODUCES HYPER-ACTIVITY,
AND IN TURN, THE HYPER-ACTIVITY
CAUSES AN INCREASE IN LPI-GPCR,
AND EVERY ARROW IS POINTING
UPWARD, IT'S A POSITIVE FEEDBACK
LOOP ALL THE WAY DOWN THE LINE
AND THIS EXPLAINS WHY EPILEPSY
CAN BE A RUN AWAY PHENOMENA AND
IT IN FACT, IT CAN REOCCUR.
AND CBD COMES ALONG, BLOCKS THE
RECEPTOR, PREVENTS THE EFFECT OF
LPI FROM ACTING ON GPR55,
ANTAGONIZES THE INCREASED LEVEL
OF GPR55, DECREASES THE LEVEL OF
EXCITATION, DECREASES SEIZURES,
PREVENTS HYPER-ACTIVITY AND
BASIC LYE PUTS OUT THE FIRE,
PUTS OUT THE REGENERATIVE
PROCESS, OR SO WE CLAIMED.
LET'S LOOK AT MORE EVIDENCE FOR
THIS BY SEEING WHETHER SEIZURES
CHANGE THE LEVEL OF GPR55 AND
THE TIME COURSE AND DYNAMICS OF
THAT.
SO IN THESE EXPERIMENTS
PERFORMED BY HELEN SCHARFMAN,
AND OTHERS COLLEAGUES IN THE NYU
PSYCHIATRY, THIS ACTIVATES
CERTAIN TYPES OF CERTAIN TYPES
OF GLUTAMATE RECEPTORS AND
PRODUCES EPILESY VERY HIPPO
CAMPALLY BASED AND IF WE INDUCE
A SEIZURE BY TREATMENT WITH
[INDISCERNIBLE] AND WAIT 48
HOURS AND THEN ASSESS THE LEVEL
OF RECEPTOR USING EITHER PC R OR
IMMUNO CYTOCHEMMISTRY, CAN YOU
SEE A VERY CLEAR INCREASE IN THE
LEVEL OF GPR 55.
WHICH IS PREVENTIVE IF YOU GIVE
CBD AT THE SAME TIME THAT YOU
GIVE KA.
SO 48 HOURS AFTER THE SEIZURE,
THERE'S A BIOCHEMICAL TRAIL,
MAKING THE SLICE MORE
SUSCEPTIBILITY TO HAVING ANOTHER
SEIZURE BECAUSE THERE'S NOR
GPR55 PRESENT.
AND THIS WAS TRUE ONCE AGAIN, IN
THE CAC1 AND 3 REGIONS IN BOTH
CASES.
THERE WAS A CLEAR ELEVATION IN
GPR55.
NOW IF IT'S THE CASE, THAT 1
SEIZURE, LAYS A TRAIL, LEAVES A
TRAIL BEHIND AND LEAVES A
PREDISPOSITION FOR A SECOND
SEIZURE, WE SHOULD BE ABLE TO
COME IN WITH A SECOND PULSE OF
KA, AND THEN LOOK TO SEE WHETHER
THAT PRODUCES A HIGHER INCIDENCE
OF SEIZURES, SO WE TREATED THE
TISSUE OF THE ANIMALS WITH A LOW
DOSE OF KA, AND 1 NOT ENOUGH TO
PRODUCE A SEIZURE AND
NONETHELESS WE SAW THAT WE CAME
IN WITH A SECOND DOSE OF KA, THE
SECOND DOSE WAS EFFECTIVE 14 OUT
OF 15 TIMES TO PRODUCE THE
SEIZURE TO THE SECOND ROUND,
THIS IS LIKE PAIRED PULSE
FACILITATION, 2 POSTS OF KA,
FITTER 1 DOESN'T DO MUCH, BUT
THE SECOND 1 DOES A LOT, NOW IF
WE TREAT WITH CBD JUST BEFORE
THE SECOND DOSE OF KA, THEN WE
PREVENT THE SEIZURES, THATTED BE
EXPECTED BUT MORE IMPORTANTLY IF
WE TREAT WITH CBD, DURING THE
FIRST EXPOSURE TO KA, THEN
LIKELIHOOD OF SEEING SEIZURES IS
CUT IN HALF.
THAT MEANS THAT THE TRAIL LEFT
BEHIND BY THE FIRST SEIZURE IS
DEPENDENT ON WHETHER CBD WAS
PRESENT, AND SO THIS FITS WITH
THE HYPOTHESIS, THAT UNCHECKED
SEIZURES, BEGET FURTHER SEIZURE,
WE TESTED THIS AGAIN BY LOOKING
NOT AT SEIZURE SUSEPTIBILITY BUT
LOOKING AT EXCITATORY MINIS TO
SEE THE SAME TYPE OF EFFECT
THERE.
WE SHOWED THE EECT OF WELL, PI
IN THE ABSENCE OF SEIZURE AND
CONTROL ISSUE ITS.
IT INCREASED THE FREQUENCIES OF
MINIS AND WE COMPARED THAT WITH
WHAT HAPPENED IN TISSUE TAKEN
FROM MICE THAT HAD BEEN DRIVEN
TO SEIZURES BY LITHIUM
PILOCARPINE, YET ANOTHER WAY OF
MAKING SEIZURES AND 1 THAT
EPILEAPTOLOGYISTS SWEAR BY
BECAUSE IT WILL PRODUCE CHRONIC
SEIZURES OVER AND OVER AGAIN AND
HERE YOU CAN SEE THAT THE EFFECT
OF LPI IS GREATLY ENHANCED, MORE
THAN DOUBLED IN SIZE AND EVEN
AFTER YOU WASH OFF THE LPI, THE
ENHANCEMENT OF MANY FREQUENCY IS
SUSTAINED.
SO THIS IS ALMOST LIKE LTP IN
THAT A SYNAPTIC INPUT IS
PROLONGED IN ITS EFFICACY.
AND SO THIS IS AN INDICATION
THAT THE CELLULAR LEVEL, THAT 1
SEIZURE LEAVES BEHIND A TRAIL
THAT MAKES IT MORE LIKELY TO SEE
A SECOND SEIZURE, AND WE SAW A
SIMILAR ACCENTUATION OF THE LPI
INDUCED DECREASE IN MANY IPSC
AMPLITUDE.
ONCE AGAIN THAT'S POST SYNAPTIC,
THIS POST IS PRESYNAPTIC BUT IN
BOTH CASES CHRONIC SEIZURES MAKE
THE EFFECT BIGGER, NOT
UNEXPECTEDLY BECAUSE THE NUMBER
OF IMRKS PR 55 RECEPTORS IS
INCREASED AND THIS EXPERIMENT
WAS DONE BY MICHAEL BASSELOT,
AND WE HAD HELP FROM AUSTIN
UNIVERSITY DOING CONTROL
EXPERIMENTS THAT WE COULD
CONTROL THIS BY CHRONIC
TREATMENT WITH CBD, SO IT ALL
FITS WITH THE HYPOTHESIS NAMELY
THAT CBD IS A VERY CLEVER DRUG
THIS IN THE WAY THAT AFFECTS
SEIZURES AND IT PREVENTS THE
POSITIVE FEEDBACK ASPECTS OF THE
CIRCUIT AND TAKES OUT WHAT IS
VERY CHARACTERISTIC OF EPILEPSY,
NAMELY THAT 1 SEIZURE, CAN LEAVE
BEHIND A TENDENCY TO HAVE
ANOTHER SEIZURE AT A LATER TIME.
NOW, SO FAR, I FOCUSED ON
SYNAPTIC TARGETS OF CBD, HOW
ABOUT ION CHANNELS AND ARE THEY
INVOFFED.
SO WE LOOKED THEN AND THIS IS
SIMON CHAIMER LAND THIS, IS 2
MAJOR CATEGORIES OF NEURON,
WE'RE NOT GOING DOWN AS FINE AS
ALL THE VARIOUS SUBTYPES, WE'RE
JUST GOING TO DISTINGUISH
BETWEEN FAST SPIKING
INTERNEURONS AND REGULAR SPIKING
INTERNEURONS THAT ARE
CHARACTERIZED BY BEING RICH IN
THE MARKER SOPHISTICATED
MATOSTAT.
THEY HAVE VERY DIFFERENT
MORPHOLOGIES, THE PV NEURONS ARE
VERY OFTEN BASKET CELLS WITH
CELL BODIES AND LOTS OF AXONAL
RAM IIVES IN THE PARAMETRAL
LAYER WHERE THEY ACT AS A DAMPER
ON PARAMETER EXCITATION, AND WE
CAN SCHEMEATIZE THEM IN TALKING
ABOUT FEET FORWARD INHIBITION
WHICH OCCURS WHEN IT FIRES A
NEURON AND SO THAT'S FEET
FORWARD IPT GREATER HIB
THORSITION, AND AFTER IT FIRES
OR IF IT FIRE SAYS, THE
SOPHISTICATED MAT'S STATIN
NEURON IS CAUSED TO RELEASE
GABBA FAR AWAY FROM THE CELL
BODY, TRANSFERRING ACTIVITIES
AND PROJECTS INPUT AND 1 LAYER
OF THE HISM HIPPOCAMPUS AND
INHIBITORY REGION IN THE
HIPPOCAMPUS.
AND THIS IS HEAVILY STUDIED BY
CHRIS Mc MAIN, KEN PELKY AND
THEIR COLLEAGUES HERE AT NIH AND
THEY'VE GONE ON TO EVEN MORE
TYPES BUT FOR NOW THIS, IS
ENOUGH TO MAKE THE POINT I'M
GOING TO MAKE THAT IS THE
[INDISCERNIBLE] WHEREAS THE
FEEDBACK INHIBITION HAS OTHER
PURPOSE, SO WHAT HAPPEN WHEN IS
YOU TREAT WITH CBD TO THESE
FORMS OF INHIBITION?
WELL, HERE'S A MORE
PHYSIOLOGICAL INPUT, A SHORT
TRAIN OF ACTION POTENTIALS AT 50
HURTS SO THE ACTION POTENTIALS
ARE 20 MILLISECONDS APART
APPROXIMATE AND WE WILL GIVE
THEM 5.
SO THIS IS TYPICAL OF A BURST OF
FIRING IN THE HIPPOCAMPUS IN
RESPONSE TO SYSTEMS NEUROSCIENCE
LEVEL ACTIVITY.
IN THE CONTROL IN THIS SLICE,
THERE MAY BE 1 SPIKE, MAYBE 0
SPIKES BUT NOT VERY MANY SPIKES
IN THE PRESENCE OF CBD, ALL THE
SPIKES ARE COMPLETELY GONE AND
ON AVERAGE, THERE IS A VERY
PROMINENT CHANGE IN THE STIMULUS
NUMBER DEPENDENTS OF WHETHER
THERE'S AN ACTION POTENTIAL OR
NOT, EXCITABILITY IS GREATLY
DECREASED IN THE PARIETAL
NEURON.
WHY DOES THIS OCCUR IN LET'S
LOOK AT USING STIEWMUE LIE THAT
ARE WEAK ENOUGH NOT TO PRODUCE
ANY SPIKES SO WE CAN LOOK AT THE
COMPOUND SYNAPTIC POTENTIAL THAT
ARISES FROM EXCITATORY
TRANSMISSION AND INHIBITORY
TRANSMISSION ALL OCCURS AT THE
SAME TIME.
IN THE ABSENCE OF CBD, WE SEE A
GRADUAL BUILD UP OF
DEPOLARIZATION, THIS IS VERY
PHYSIOLOGICAL, THIS IS WHAT
WOULD MAKE THE NEURON REACH THE
THRESHOLD, ONLY WE'RE NOT QUIT
HITTING THRESHOLD HERE SO WE CAN
ANALYZE THE SYNAPTIC EVENTS IN
RESPONSE TO CBD, THE
NETDEPOLARIZATION IS STRONGLY
DECREASED, CONSISTENT WITH THE
LOWERING OF THE NUMBER OF ACTION
POTENTIALS AND THIS IS GROUP
DATA SHOWING THE SAME THING.
NOW WHAT HAPPENS IF WE KNOCK OUT
THE RECEPTOR?
WELL, IN THE ABSENCE OF THE
RECEPTOR THIS EFFECT IS
COMPLETELY GONE.
WHAT HAPPENS IF WE MIMIC THIS
WITH AN ARTIFICIAL AGENT, NOT A
PH ITOCANNABOID OF THE RECEPTOR
BUT IT'S SELECTIVELY BEEN
ENGINEERED FOR THE RECEPTOR IT'S
CALLED CID AND AT A LOW
CONCENTRATION THIS MIMICS THE
EFFECTS OF CBD, IN PRESENCE OF
CID, WE APPLY CBD, AND NOW CBD
HAS NO ADDITIONAL CENTER FOR
EXCELLENCE ON AGING.
SO THE EFFECT OF CBD HAS BEEN
OCCLUDED BY THE PRESENCE OF CID,
ONCE AGAIN RECONFIRMING THE IDEA
THAT IS DUE TO THE GPR55
RECEPTOR.
WHETHER YOU KNOCK OUT THE
RECEPTOR ASK ILLUMINATE THE
EFFECT OR STIMULATE THE RECEPTOR
USING CID AND GET THE FULL
EFFECT, EITHER WAY, IT'S VERY
CONVINCING EVIDENT THIS EFFECT
ON EXCITABILITY AND SYNAPTIC
TRANSMISSION IS GPR 55
DEPENDENT.
THE KEY EXPERIMENT IS TO COMPARE
THE RESULTS OF EXCITATORY
TRANSMISSION ALONE OR EXCITATORY
TRANSMISSION WITH GABBA
RECEPTORS BLOCKED SO NORMALLY WE
HAVE BOTH GLAITA MATE RECEPTORS
AND GABBA RECEPTORS, IF WE BLOCK
GABBA RECEPTORS NOW ONCE AGAIN
CBD GOES COMLITELY AWAY.
THAT MEANS THIS WAS NOT AN
EFFECT ON THE EXCITATORY TRAN
MISSION, IT WAS AN EFFECT ON
INHIBITORY TRANSMISSION THAT
COUNTERACTED THE EFFECT OF THE
EXCITATORY TRANSMISSION, SO WE
SEE A COMPLETE BUILD UP WITHOUT
ANY IPSCs, SO THIS TELLS US
THAT IT'S MEDIATED BY THE GABBA
SERNLGIC SYSTEM AND NOT THE
GLUTEA MA TERNLGIC SYSTEM, SO
WHICH NEURON IS RESPONSIBLE FOR
THIS.
THE OBVIOUS POSSIBILITY IS THAT
PV NEURONS WHICH MEDIATE FEET
FORWARD INHIBITION ARE WRONG.
SO TO STUDY THIS WE CAN NOW
RECORD FROM PC NEURONS AND SO
THIS IS NOT THE SPIKING IN THE
EXCITATORY NEURONS, AND THE
EXCITATORY NEURONS AND LOW AND
BEHOLD, CBD, GREATLY INCREASES
THE FIRING, MAYBE 3 FOLD, SO THE
PV NEURONS INSTEAD OF FIRING
EVERY ONCE IN A WHILE FIRE
ALMOST NEARLY ALL THE TIME AND
THIS EFFECT IS MIMICKED BY CID,
SHOWING ONCE AGAIN THAT IT'S DUE
TO GPR 55.
SO, NOW WE HAVE AN EFFECT OF CBD
ON THE INHIBITORY NEURON ITSELF,
NOT ONLY ON THE INHIBITORY
NEURON BUT THE VERY INHIBITORY
NEURON THAT IS MOST
STRATEGICALLY LOCATED AND PLACED
IN THE CIRCUIT TO QUELLCH ACTUAL
POTENTIAL PROPAGATION.
HOW ABOUT SST NEURONS, WE SEE
THE OPPOSITE, INSTEAD OF THESE
NEURONS FIRING MORE TO INCREASE
INHIBITION, THEY ARE FIRING LESS
AND THAT EFFECT IS ALSO MIMICKED
BY CID.
SO THE 2 INTERNEURON TYPES ARE
AFFECTED IN OPPOSITE DIRECTIONS.
WHAT IS THIS DUE TO?
IS IT DUE TO THE EXCITATORY
TRANSMISSION OR DUE TO INTRINSIC
EXCITABILITY, BECAUSE REMEMBER
WE'RE FIRING SPIKES HERE, WE'RE
DRIEBING THE PROBABILITY OF
SPIKES, SO THIS SHOW WHAT IS
HAPPENS IF YOU DEPOLARIZE, USING
THE CURRENT PATCH CLAMP
ELECTRODE BYPASSING INAPTIC
TRANSMISSION TO MAKE THE NEURON
FIRE SPIKES, HERE'S A FAST
SPIKING INTERFER NEURON AND
RESPONSE TO CBD, THE ACTUAL
POTENTIAL FETAL COMPARTMENT
FERENCEY GOES UP SIGNIFICANTLY,
PARTICULARLY PROMINENTLY AT
SMALL CURRENT INJECTIONS, SO CBD
ENHANCES THE INTRINSIC
PROPERTIES OF FIRING OF PV
NEURONS, IT HAS THE OPPOSITE
EFFECT ON SST NEURON.
SO THIS IS VERY CLEVER BECAUSE
IT'S ENHANCING THE EXCITABILITY
OF THE VERY FIEWRON THAT IS BEST
POSITIONS TO MEDIATE FEET
FORWARD INTERMISSION EMPLOY NOW
WE TESTED SODIUM EFFECTS OF
THIS, ON THE HYPOTHESIS, THIS
MIGHT INVOLVE ENHANCEMENT OF
SODIUM CARBONS AND THE
HYPOTHESIS WOULD BE THAT CBD
COULD PREVENT ENDOGENOUS DOWN
REGULATION OF SODIUM CURRENTS
MEDIATED BY CONVENTIONAL
SIGNALING PATHWAYS, STUDIED BY
[INDISCERNIBLE] AND JIM
[INDISCERNIBLE], AND IF THERE
WAS A TONIC AMOUNT OF LPI DOWN
REGULATING THE SODIUM CURRENT,
THEN CBD BY INTERFERING WITH THE
SIGNAL INDUCTION COULD MAKE A
BIGGER SODIUM CURRENT OR MORE
FIRING.
AND BY LOOKING AT THE COMPOUND
EBST, THAT A SYNTHESIZED
STIMULATOR OF NAV 1.1, THE
SODIUM CHANNEL THAT'S IN THE
ALBUT MIN NEURONS WAS ABLE TO
MIMIC THE EFFECTS OF CBD AND IN
FACT CONCLUDE THE EFFECTS OF CBD
AND THIS WAS MIRRORED BY ANOTHER
COMPOUND, AA43729, ONCE AGAIN
MIMICKING THE EFFECTS OF CBD.
SO WE THINK WE ARE ON TO WHAT
THE BASIC MECHANISM OF HOW CBD
MIGHT ACT AND WE THEN TRY TO
TEST WHAT THE TIMING OF THE
INPUT NEEDED TO BE BY DOING AN
OPTICAL IMAGES O GENETIC
EXPERIMENT EXPRESSING CHANNEL
ROUGH ATOM DOBSIN IN PV NEURONS
AND SHOWING WE CAN MIMIC THE
EFFECT OF CBD WITHOUT ANY DRUG
AT ALL BY SHINING LIGHT ON THE
TERMINALS OF THE PV NEURONS ON
TO THE CELLS AND YOU CAN SEE
REGARDLESS OF WHETHER YOU HAVE
WEAK STIMULATION, STRONG
STIMULATION, THE FEET FORWARD
INHIBITION MIMIC THE EFFECT OF
CBD, THIS IS NOT PRODUCED IF
WE'VE SHOWN A VERY GENERAL AND
TIME INDISCRIMINATE STUDY GLOW
OF LIGHT, THEN WE SAW THE EXACT
OPPOSITE OF THIS EFFECT.
AND SCOTT OWEN WHO IS WORKING
WITH THE INTRAMURAL DIRECTORS,
SON, RELATIVE, WAS THE 1 WHO
DESCRIBED THIS FOR PV NEURONS
USINGOXITOSEIN, THE GENERAL
LIGHT DOES WHATOXITOSEIN DOES,
SO WHAT YOU NEED IN ORDER TO
PREVENT THE THROUGH PUT OF
TRANSMISSION IS PRECISELY TIMED
ACTIVATION OF THE PV NEURONS,
THE RIGHT PLACE AND AT THE RIGHT
TIME, RIGHT AS YOU WOULD EXPECT
FROM FEET FORWARD INHIBITION, SO
THE CONCLUSION IS THAT CBD IS A
VERY CLEVER DRUG.
IT DAMPENS THE PROPAGATION OF
HIFREQUENCY ACTIVITY IN THE
HIPPOCAMPUS BY INCREASING PV
INTERNEURON FIRING AND
CONCOMITANTLY DECREASING THE
FIRING OF SST INTERNEURONS,
MAINTAINING SOME SORT OF PV SST,
I WANT TO USE THE WORD BALANCE,
DON'T WANT TO USE THE WORD
BALANCE BECAUSE IT REMINDS ME OF
EI BALANCE BUT ANYWAY, IT'S SOME
KIND OF HARMONY BETWEEN THE 2
TYPES OF INTERNEURONS, THE 1 YOU
NEED TO FIRE IS THE 1 THAT'S
MADE THE FIRE AND THAT DAMPENS
SPIKE TRANSMISSION BY PATHWAY
GIVES RAMETTAL CELLS AND THE SST
NEURONS ARE NOT ALLOWED TO
INHIBIT THE PV NEURONS O SO THAT
MAKES THE EFFECT EVEN STRONGER.
WE CAN MIMIC THE EFFECT BY
ACTIVATING SODIUM CHANNELS AND
THEY MIMIC AND INCLUDE CBD
CIRCUIT ACTION.
AND THE TIMING IS IMPORTANT
BECAUSE THAT'S WHEN YOU NEED THE
INHIBITORY INPUT TO PREVENT
FIRING AND THESE EXPERIMENTS
WHICH ARE DONE WITHOUT ADDED LPI
ARE BETTER SUITED TO THE
NEUROPSYCHIATRIC INDKAIGS THAT
CBD ARE USED FOR BECAUSE PEOPLE
WHO ARE TAKING CBD FOR WHATEVER
AILS THEM, SOMETIMES
INTELLIGENTLY AND SOMETIMES
BLINDLY, THEY'RE NOT EPILEPTIC
PATIENTS THEY'RE JUST PEOPLE
WORKING WITH THE BRAIN'S OWN
LEVEL OF LPI, SO THE VIRTUE OF
HAVING AN ENDOGENOUS SYSTEM IS
THAT IT'S MODULATING THE PROPERS
OF CIRCUIT AND REGULATING HOW
INFORMATION GETS TRAN FERRED AND
IT PROVIDES A STRATEGIC TARGET
FOR CBD ACTION.
NOW LET ME CHECK HOW I'M DOING
ON TIME, AND I'D LIKE TO SEGUE
TO THE LAST PART OF MY TALK AND
BELIEVE ME, THEY'RE NOT 50% MORE
SLIDES, THERE ARE ONLY MAYBE 33%
MORE SLIDES AND I THINK I HAVE
ENOUGH TIME TO DO THIS.
>> THIS MIGHT BE A GOOD MOMENT,
I'M OVER HERE, THE OTHER
DIRECTION, I JUST WANT TO,A
NOUNS, YEAH, SORRY.
OUR SEND LIVE FEEDBACK BUTTON IS
NOT WORKING SO ANYONE WATCHING
IT REMOTELY AND THERE ARE A LOT
OF PEOPLE WATCHING REMOTELY,
SEND E-MAIL TO THE WAS OFFICE,
WALSOFFICE@OD.NIH.GOV.
>> THANK YOU, WE WANT ALL THE
FEEDBACK WE CAN GET.
AND WE'RE HAPPY TO GET IT
WHETHER IT'S IN A CHAT OR E-MAIL
OR ANY FORM.
E-MAIL MIGHT BE EVEN BETTER
BECAUSE I CAN PASS IT ON TO MY
COLLEAGUES WHO ARE ALWAYS
ELECTRICKING FOR WAYS
TO--LOOKING FOR WAYS OF
IMPROVING OUR WORK.
SO I DRIEB THIS VICIOUS CYCLE,
THIS POSITIVE FEEDBACK WHERE
SOMETIMES THE LIGAND AND ALWAYS
THE RESUPPORTOR THAT TRIGGERS AN
INN CREASE IN EXCITATION AND
DISCIPLINARY CREASE IN
INHIBITION IS ENHANCED BY
HYPER-ACTIVITY, A POSITIVE
FEEDBACK LOOP THAT'S INTERRUPTED
BY CBD, THIS IS AT THE RECEPTOR,
SYNAPSE LEVEL AND THAT CIRCUIT
LEVEL, THE NUMBER OF SPIKES THAT
CAN GET THROUGH, IS ENHANCED BY
A GREAT ENHANCEMENT OF FEET
FORWARD INNOVATION, AND
CONCOMITANT FEET FORWARD AND
BACK INHIBITION, AND TOGETHER
THIS DECREASES THE NET EPSB, AND
DECREASED SPIKE THROUGH PUT.
SO BOTH AT THE SYNAPTIC LEVEL
AND AT THE CIRCUIT LEVEL AND
NEURONAL FIRING LEVEL, CBD IS A
VERY EFFECTIVE AGENT IN
MODIFYING IMPULSE TRAFFIC AND
INFLAMMATION FLOW THROUGH THE
BRAIN.
I WOULD LIKE TO CLOSE WITH
MEDICAL DETECTIVE STORY ABOUT
KIDS.
WHICH WAS HEAD LINED HERE IN
SCIENCE AS SEQUENCING MYSTERIOUS
CASES, YIELDS MUTATIONS LINKED
TO SEIZURES AND ARRHYTHMIAS, AND
THIS GOES BACK TO MICHELLE'S
VERY KIND AND INTRODUCTION ABOUT
THINGS THAT INTEREST US AND I
THINK SHOULD INTEREST PEOPLE AT
NIH.
SO WHAT IS SUDC, WELL, IT'S SIDS
IN OLDER KIDS, THE ONLY
DISTINCTION BETWEEN THESE 2
SYNDROMES IS WHETHER THE CHILD
IS LESS THAN 1 OR OVER 1 AND
MOST OF US WOULD BELIEVE THERE'S
NO WATERSHED AND MECHANISM THAT
STOPS AT 1 YEAR AND THEN
BEGENERATEDS AT 12 MONTHS.
IN FACT, IT'S CONTINUUM AND
GENETICS IS CONFIRMING THIS
IDEA.
ALLOWING FOR ITS PREVALENCE MAY
BE NOT AS LARGE AS SIDS BUT
STILL A SUBSTANTIAL SUBTRACTION
OF SIDS PATIENTS.
IN FACT THE RATIO COULD BE
SOMETHING LIKE 3 OR 4 TO 1.
THE NUMBER OF CASES IS LESS FOR
SUDC, BUT THE FUNDING THE RATIO
IS PRACTICALLY INFINITE AND THE
RATIO OF PUBLICATIONS IS ALSO
EXTREMELY HIGH.
SO IF YOU NOMPLALLIZE THINGS
ACCORDING TO THE NUMBER OF
PATIENTS, SUDC IS
UNDERREPRESENTED.
SO HOW DO WE APPROACH THIS,
WELL, SOMEONE WHO'S BOTH A
RESEARCHER, AND A MOM WHO
SUFFERED A TRAGEDY EARLY IN THE
FAMILY STARTED A CONSORTIUM
CALLED THE SUDC REEJ TREE ASK
RESEARCH COLLABORATIVE AND SHE
TEAMED UP WITH OUR RESEARCH
PATHOLOGIST AND THEY REVIEWED
THE KIDS DEATH, MADE AN ANALYSIS
AND MADE A CASE REPORT IN
CLINICAL JOURNALS TO USE THIS
INFORMATION TO IESHES DEPTIFY
AT-RISK INDIVIDUALS AND THEY
TEAMED UP WITH PEOPLE IN
PEDIATRICS WITH COLUMBIA IN THE
GENETIC BACKGROUND AND PEOPLE
OUT IN THE COMMUNITY AT THE MAYO
CLINIC WHO WERE INVOLVED IN
DOING THE DETECTIVE STORY ABOUT
FIGURING OUT WHAT GENETIC AND
ENVIRONMENTAL FACTORS TRIGGER
SUDC, AND THEY COLLECTED UP TO
130 FAMILY TRIOS WHERE THERE IS
A DECEASED CHILD, BOY OR GIRL,
PRETTY MUCH 50/50 AND WOPARENTS
WHO ARE WILLING IN THE WAKE OF
THE TRAGEDY TO GIVE BLOOD
SAMPLES AND WHERE THEIR BLOOD
SAMPLES BEING BE COMPARED WITH
THAT OF THE CHILD.
AND SO THAT'S CALLED TRADITIONAL
ANALYSIS OF TRIOS, MOM, DAD AND
SON OR DAUGHTER AND THE CODING
AXONS OF THE INDIVIDUALS ARE
COMPARED, AND SO THIS IS A
COLLECTION AND ALMOST ALL KIDS
DIED IN THEIR SLEEP, AND VERY
FEW WHO ARE AWAKE.
I WILL COME BACK TO WHAT
HAPPENED WHILE THEY WERE
SLEEPING LATER.
IN MOST OF THE CASES, MAYBE 40%,
THE KIDS HAD SOME SORT OF
SEIZURE HISTORY, FEB RILE
SEIZURE, SOME OTHER TYPE OF
SEIZURE, SO THAT IMMEDIATELY
MAKING YOU THINK ABOUT BRAIN
CIRCUITS, AND THIS WORK WAS PUB
LIBRARY FOUNDATIONED A COUPLE
YEARS AGO.
SO THE EXOHM SEQUENCING OF THESE
TRIOS LED TO TABLES LIKE THIS,
AND THIS IS JUST THE FRAGMENT OF
A LONG LIST OF TABLES WHERE THEY
WERE ABLE TO COME UP WITH
CANDIDATE MUTATIONS THAT MIGHT
CONTRIBUTE TO THE DISORDER.
AND SOMETIMES THEY WERE
FORTUITOUSLY FOUND MUTATIONS
THAT WERE ALREADY DRIEBED IN THE
DATABASE AS POTENTIALLY BEING
PATHOGENIC.
THIS IS A DESCRIPTION VARIED ON
PRODUCED BY A DATABASE AND THEY
HAVE NUMBERS, THEY HAVE VARIOUS
WAYS OF CATALOGING THEM, AND SO,
WE USE THIS AS A WAY OF
EDUCATING OURSELVES INTO HOW
GENETICISTS APPROACH THINGS
TAKING THIS AS A TEST CASE, A
TRAGIC TEST CASE BUT 1 THAT
OFFERED OPPORTUNITIES AND WE
COMPARED THE EFFECTS OF
SYNONYMOUS VARIANTS AND
NON-SYNONYMOUS VARIANTS.
VARIANTS THAT DID NOT CHANGE THE
AMINO ACID THAT WAS CODED FOR OR
NON-SYNONYMOUS VARIANTS THAT
COULD FEBRUARY EXAMPLE CHANGE A
DPLI SEEN TO A SERIES POINTSINE
AND MATT HALVERSON, A GREAT
GENERATEDETTIST, DID THE
ANALYSIS, THE SYNONYMOUS
VARIANTS PROVIDED GREAT CONTROL,
BECAUSE THERE'S BEEN A CHANGE IN
THE DNA BUT NO CHANGE IN THE
PROTEIN, WE'RE NOT EXPECTING
THESE TO BE LETHAL, IN FASTBALL
WE FOUND THAT THE SUBSET OF
YEENS WHICH ARE LABELED CARDIAC
EPILEPSY YEANS NOT
DISTINGUISHING BETWEEN
ELECTRICAL ACTIVITY, BETWEEN THE
HEART OR THE BRAIP, WE LUCHED
THEM ALTOGETHER AND WE HAD 137
YEENS AND WE LOOKED TO SEE WHAT
WERE THE ODDS THAT YOU WOULD
FIND A DE NOVO MUTATION IN THIS
SUBSET OF YEEN ANDS IN FACT, WE
FOUND THAT THE NON-SYNONYMOUS
VARIANTS WERE IN THESE
PARTICULAR GENES WERE
SIGNIFICANTLY AFFECTED BY
THE.WHERE ARE 032 LEVEL, SO
THESE YEENS ARE A NATURAL PLACE
TO LOOK BECAUSE THEY SHOW THE
SIGNS OF WHAT YOU EXPECT IF THEY
PRODUCE DISEASE, THE CONTROL OF
HAVING THE SYNONYMOUS VARIANCE
IS VERY SATISFYING BECAUSE THESE
WERE NOT SIGNIFICANTLY CHANGED
AND AS YOU WOULD EXPECT BECAUSE
THESE ARE NOT CHANGING THE
PROTEIN.
NOW LET'S TAKE THESE GENES, TAKE
THE MUTATION, WE FOUND IN THE
SUBSET OF 125 INDIVIDUALS ASK
RUN THESE AGAINST OTHER DECS OF
KIDS, CANCERS, NOT EXPECTING
ANYTHING, OTHER RARE DISEASES,
ASTHMA, DEVELOPMENTAL DELAY AND
ALL THESE PLACES, THERE WAS NO
ENRICHMENT OF THESE, THE GENES
IN THESE DISEASES, IN OUR
SUBSET, BUT IF WE LOOK AT AUTISM
WHERE THERE ARE 700?
SEVENTEEN GENES IMPLICATED BY
THE SIMONS FOUNDATION OR OTHER
ORGANIZATIONS, WE EXPECTED TO
SEE AN OVERLAP OF ABOUT 4 AND
HALF IN 4 AND HALF CASES BUT WE
SAW IN 11 CASES THERE WAS AN
OVERLAP AND AN ENRICHMENT WITH A
HIGHLY SIGNIFICANT P-VALUE
TELLING US THAT IN THE GENETIC
ALONE THAT THE DISEASES THAT
COME UP IN SUD ARE HIGHLY
RELEVANT AND VICE VERSA AND THIS
IS THE TYPE OF ANALYSIS YOU CAN
DO WHEN YOU'RE NOT ACTUALLY
LOOKING FOR SOMETHING, YOU DON'T
HAVE A PREDETERMINED HYPOTHESIS,
YOU'RE JUST TAKING WHAT IS GIVEN
TO YOU BY PEOPLE COMING INTO THE
REGISTRY.
SO THAT'S THE CONCLUSION AND
CONFIRMS WHAT MANY PEOPLE HAVE
ALWAYS BELIEVED THAT AUTISM AND
EPILEPSY HAVE A STRONG OVERLAP
EMPLOY NOT AUTISM AND HEART
DISEASE OR CARDIAC
ARITHMETRICSAS, AUTISM AND
EPILEPSY.
JOHN WANG TOOK A DIFFERENT
APPROACH, HE LOOKED AT THE
YEERNS THAT WERE CHARACTERIZED
AND PUT THEM INTO PROTEIN,
PROTEIN INTERACTION NET WORKS SO
PEOPLE CATALOG THE WAY PROTEINS
INTERACT WITH PARTNERS BY DOING
REAL HONEST TO GOODNESS
BIOCHEMISTRY AND THEY FIND THAT
THE GENES WE FIND AFFECTED BIKE
[INDISCERNIBLE] RYR2 AND 2 CASES
EACH AND CAL-MODUE LYNN WERE ALL
PART OF A SOCIAL NETWORK, A
NETWORK BETWEEN ENCODED PROTEINS
THAT PHYSICALLY IRPT ACT WITH
EACH OTHER AND FOR SYNONYMOUS
VARIANTS THIS ENRICHMENT VALUE
WAS PRACTICALLY A CHANCE WHEREAS
IN THE NON-SYNONYMOUS PROTEIN
VARIANTS, THE LIKELIHOOD OF
THERE BEING THIS MUCH
PROTEIN-PROTEIN INTERACTION IS
SIGNIFICANT.
SO NOT ONLY DO THE PROTEINS
CAUSE LETH AGALLITY BUT THEY
CAUSE IT IN PROTEIN, PROTEIN
ISHT WORKS.
THIS IS NOT ONLY TRUE OF A
CLUSTER, KNOWN FOR ITS ROLE IN
CALCIUM REGULATION, IT WAS ALSO
TRUE IN A CLUSTER KNOWN FOR
NEURAL DEVELOPMENT WHICH IS VERY
INTERESTING BECAUSE WE'RE NOT
EXPECTING THIS, WE WERE LOOKING
FOR SOMETHING THAT WOULD ACT
ACUTELY, THIS SAYS THAT SOME OF
THE GENES WE FOUND CLUSTERED
TOGETHER BECAUSE OF PROTEIN
INTERACS THAT WERE IMPORTANT IN
DEVELOPING THE NERVOUS SYSTEM.
SO SO THAT ARE THE EFFECTS OF
THESE GENES, SOME OF THEM WERE
ALREADY KNOWN, THE MUTATION AND
CAL-MODUE LYNN CAUSES SLOWER
CALCIUM ACTIVATION AND IS LETHAL
WITH BETA MINERGIC STIMULATION
IN MICE.
THE [INDISCERNIBLE] MUTATION,
CAUSED THE CHANGE IN THE CALCIUM
TRANSIENT AND A SLOWING OF THE
CALCIUM TRANSIT, I'M NOT SURE
WHAT WELL MEANS BUT THERE'S A
DEFINITE PHENOTYPE IN CALCIUM
RELEASE IN CELLS AND FINALLY, 1
OF THE OTHER MUTATIONS WE FOUND
IN OUR DATA SET, HAD BEEN
PREVIOUSLY IMPORTANT IN A
SYNDROMIC FORM OF AUTISM CALLED
TIMOTHY SYNDROME WHICH IS
CHARACTERIZED BY LONG HEART
BEATS AND TENDENCYS TO
ARRHYTHMIAS.
WE CHARACTERIZE OURSELVES AND
THIS IS GARY L. GRANT WITH THE
L-TYPE OF THE CALCIUM CURRENT
AND SHE FOUND THAT THERE FTION A
SIGNIFICANT PROLONGATION OF THE
TIME COURSE OF THE CURRENT IN
EITHER THE 2 MUTATIONS WHICH
APPEARED IN OUR DATA SET, THIS
IS A COMMON FEATURE OF COMMON
MUTATIONS THAT CAUSE ARRHYTHMIAS
AND CHANGES IN NERVOUS SYSTEM AS
WELL, HIGHLY SIGNIFICANT
INCREASE AND HOW MUCH THERE IS
LATE IN THE VOLTAGE AND
ACTIVATION IS GREATLY SLOWED BY
THE MUTATIONS.
SO PUTTING IT ALING IT, WE THINK
THAT LONGER CHANNELS CREATE
LONGER CALCIUM FLUX AND LONGER
CALCIUM FLUX CAUSES LATER
DEPOLARIZATIONS THAT ARE A
TEXTBOOK EXAMPLE OF HOW YOU CAN
PRODUCE LETHAL ARRHYTHMIAS AND
IT'S BOTH AMUSING AND SAD FOR ME
TO REALIZE I WORKED ON THIS PER
A BETTER PART OF 3 DECADES GOING
BACK TO MY WORK ON HEART TISSUE
THAT MICHELLE ANTWON ALLUDED TO.
MEAN WHILE A GROUP IN BUSTON'S
PUBLISHED SHORTLY AFTER WE DID,
INDEPENDENTLY, DIFFERENT DATA
SET OF PATIENTS AND CAME UP WITH
A VERY SIMILAR CONCLUSION THAT
ION CHANNELS AND OTHER PROTEINS
INVOLVED ARE INVOLVED AND
THEY--YOU COULD TAKE THE 2 DATA
SETS TOGETHER AND THEY WOULD BE
BOTH THE THEN OVERLAP AND THE
EXTENSION OF THE WORK AND
BECAUSE THIS IS A RELATIVELY
RARE DISEASE, IT'S AN EXAMPLE
WHERE SCIENTIFIC COOPERATION
COULD BE VERY FRUITFUL.
JUST BECAUSE HEART IS AN OBVIOUS
POSSIBILITY DOESN'T MEAN THE
BRAIN IS NOT INVOLVED BECAUSE
THE VERY SAME PROTEINS, ARE
THOUGHT TO BE INSTRUMENTAL IN
EXCITATION TRANSCRIPTION
COUPLING AND OTHER SIGNALING
ABNORMALITIES IN DIFFERENT CNS
DISORDERS LIKE SEIZURES,
MIGRAINE AND AUTISM, AND IN FACT
THE CALCIUM CHANNEL CONVEYS A
CONFIRMATIONAL SIGNAL THAT
DOESN'T NEED CALCIUM INFLUX JUST
BY THE PROTEIN CONFIRMATION THAT
COULD SIGNAL TO THE
[INDISCERNIBLE] RECEPTOR OR
OTHER PROTEINS LIKE KMK2 AND
THEREBY PRODUCE ABNORMAL
ELECTRICAL ACTIVITY.
SO THE JURY IS TILL OUT WHETHER
IT'S CARDIAC DISEASE OR NEURONAL
DEC OR BOTH, THE HEART DISEASE
COULD KILL YOU BECAUSE OF
ARRHYTHMIA, THE NEURONAL DISEASE
COULD CAUSE EPILEPSY CAN AN
EFFECT ON BREATHING AND LAURA
AND [INDISCERNIBLE] HAVE
PUBLISHED A PAPER IN NEUROLOGY
TAKING VIDEOS OF KIDS IN THEIR
BEDS RECORDED AS PART OF THE
NORMAL SURVEILLANCE THAT PARENTS
IMPOSE AND AFTER CAREFUL VIEWING
BY CLINICIANS, INDEPENDENT
REVIEWS, THEY CONCLUDED THAT
THERE WERE A SUBSTANTIAL NUMBER
THAT SHOWED TERMINAL
CONVULINGSS, EXTREMELY SAD FOR
THE FAMILIES BUT INFORMATIVE FOR
HOW WE CAN UNDERSTAND AND COPE
WITH THE DISEASE AND PREVENT AND
ULTIMATELY AND FIND A WAY OF
PREVENTING IT AND THESE ARE
PICTURES OF 3 OF THE KIDS WHOSE
ROTATIONS ARE ACTUALLY KNOWN.
THERE ARE ENOUGH OF MUTATIONS SO
THERE'S NO PROBLEM
CONFIDENTIALITY AND THESE
PICTURES WERE PROVIDED BY THE
FAMILIES AS PART OF THEIR FERVOR
TO TRY TO GAIN MORE PUBLIC
SUPPORT FOR THE IDEA OF STUDYING
THIS STUDYING
THIS DEVASTATING DISORDER.
SO IN CONCLUSION, I WILL DO IT
IF 4 MINUTES, WE'VE BEEN
FOCUSING ON RARE VARIANTS WHO
HAVE AN EXTREME OR AUTOSOMAL
DOMAIN NABT EFFECT IN CAUSING A
FATALITY AND THESE HAVE HUGE
EFFECT SAYSS BUT OBVIOUSLY
THEY'RE VERY RARE IN THE GENERAL
POPULATION.
HOW ABOUT THE BIGGER PUBLIC
HEALTH PROBLEM OF OTHER VARIANTS
WHICH CAUSE OTHER DECS,
NEUROPSYCHIATRIC DECS
PRINCIPALLY WHICH ARE EXTREMELY
COMMON AFFECTING A LARGE NUMBER
OF PEOPLE BUT WHERE THE
INDIVIDUAL VARIATION CAUSES A
SMALL EFFECT.
AND I WILL SKIP THROUGH THIS AND
SAY, WE KNOW A LOT ABOUT THE
ETIOLOGY OF THESE RARE
MUTATIONS, HOW CAN WE STUDY THE
POORLY UNDERSTOOD 1S AND 10 IS
WE TEAM UP WITH IMENET CYSTS WHO
TELL US IN AN UNBIASED WAY THAT
THERE'S A STRONG ASSOCIATION
WITH THE CACNA1C GENE, THE VERY
GENE THAT CAUSES AUTISM, OR
CAUSES SUDC, THAT YEEN IS ALSO
IMPORTANT IN OTHER DISEASES THAT
AFFECT MILLIONS POOF INCLUDING
BIPOLAR DISORDER AND WHERE IT'S
SECOND IN GETTING TO THE TOP OF
THE CHARTS.
THE SAME LOCUST HAS BEEN
ASSOCIATED WITH MAJOR
DEPRESSION, ADD, AND ASD, SO
THIS SEEMS THIS IMPORTANT
PROTEIN IS NOT ONLY IMPORTANT
FOR NORMAL FUNCTION BUT IT'S
IMPORTANT FOR DISEASE AS WELL.
TO BE VERY BRIEF, THE GENOME IS
MOSTLY NONCODING DNA, THE DNA IS
ASSEMBLED BY SPLICING WHICH PUTS
TOGETHER MATURE TRANSCRIPT WHICH
GOES OUT OF THE CYTOPLASM AND IS
USED TO ENCODE PROTEIN, HOWEVER,
THAT--THOSE INSTRUCTIONS ONLY
USE ABOUT 1% OF THE REAL ESTATE
IN OUR GENOMES.
ONE% OF OUR GENOMES ARE ACTUALLY
ENCODING GIVING SPECIFIC
INSTRUCTIONS TO MAKE THE AMINO
ACID SEQUENCE IN THE PROTEIN,
THE REST OF THE GENOME IS
NONCODING, WHAT'S IT THERE FOR?
TO REGULATE THE ENCODING PARTS
EITHER BY MAKING THE CHROMATIN
INACCESSIBLE OR ACCESSIBLE OR
ALLOWING VARIOUS TRANSCRIPTION
FACTORS TO BIND AND TO SKIP THE
TO THE VERY END, WE ARE FOCUSING
ON CACNA1C ISSUE BECAUSE ABOUT
HALF THE GENE IS MADE UP OF 1
ENTRON AND THAT HARBARS THE
SNPs ARE IF SITS FRENNIA AND
OTHER MAJOR PSYCHIATRIC DISEASES
AND EVEN IF YOU NARROW IT DOWN
TO BASIAN ANALYSIS THAT ARE
THOUGHT TO BE CAUSAL, YOU STILL
HAVE SEVERAL INDIVIDUAL
VARIATIONS BUT WE'RE TAKING AN
APPROACH TO THIS LED BY INTREPID
COLLABORATOR NAMED SEAN WANG AND
WE'RE TRYING TO MAKE ARTIFICIAL
ALLELES WHICH CONTAIN ALL RISK
OR ALL PROTECTIVE VARIANTS OF
THIS DOING IT BY CLEVER
LEVERAGING OF BIOLOGICAL
TECHNIQUES.
WE HOPE TO PUT THESE INTO HUMAN
CELLS TO STUDY ORGANOID ANDS PUT
THEM INTO MICE, TO STUDY
BEHAVIOR AND HOPEFULLY THEN WE
WILL BE ABLE TO USE THE SAME
TYPE OF APPROACHES WE'VE USED
FOR MANYIERS TO ANALYZE THE
BASIS OF THE DEC, BY LITERALLY
COMPARING WHAT HAPPENS WHEN YOU
HAVE A LAYER THAT KEEPS YOU
PERFECTLY PROTECTED TO
SCHIZOPHRENIA, OR PERFECTLY
SUSCEPTIBLE TO IT.
WE'RE NOT EXPECTING ALL THE RISK
FACTORS TO ADD BECAUSE SOME OF
THEM ARE LINKED TO EACH OTHER
BUT WE EXPECT CUMULATIVELY YOU
SHOULD HAVE AN EFFECT THAT YOU
SHOULD BE ABLE TO SEE BEHAVIOR
CHANGE IN A MOUSE MODEL OR
EXPRESS EFFECT IN A HUMAN
DERIVED CELL.
SO TO CONCLUDE, NEURONS PROVIDE
CONNECTION AND FUNCTION AND SUDC
IS A CLEAR EXAMPLE OF THAT.
WE HAD NO IDEA WHERE TO LOOK,
GENIX TOLD US WHERE TO LOOK BUT
WHEN WE LOOK WE FIND THAT THE
FUNCTION APPROACHING THESE IS
DRASTICALLY AFFECTED.
THIS FUNCTION DIFS PERSPECTIVE
ON THE HEALTHY BRAIN, EVEN IN
THE HEALTHY BRAIN, THERE'S LPI,
IT'S BEING MADE ALL THE TIME AS
A RESULT OF THE ACTION OF PLA 2
AND NLP A IT1 AND IT'S FORTUNATE
SETTING IN EI RAT WROA EVEN
UNDER BASAL CONDITIONS SO WHEN
PEOPLE TALK ABOUT EI BALANCE
WHICH IS 1 POINT ALONG AN EI
RATIO CURVE AND RATIO ACCESS,
THAT'S AFFECTED BY LPI, WHO
CONTROLS THE CONTROLLERS, LIKE,
THE EXCITATORY AND INHIBITORY
SYNAPSES, CALCIUM CHANNELS, GENE
EXPRESSION STUDIES CAN FOCUS ON
THE SIGNALING PROTEINS THAT LIE
UP STREAM OF THOSE SIGNALING
PROTEINS, PROTEINS LIKE GPR 55.
THE LPI GPR 55 SYSTEM FOLLOWS
THE PRECEDENT OF ENDOKACCT
NABBOIDS AND THEIR RECEPTOR EC1
STUDIED HERE AT NIH AMONG MANY
OTHER PLACES AND DESPITE ALL
THIS INTENSE EFFORT, THE LIPID
SIGNALING STILL HOLDS MANY
MYSTERIES, BECAUSE THE LIPID IS
NOT AS EASY TO STUDY AS A
NEUROTRANSMITTER POLARIZED KIEWL
WHICH IS EASILY--MOLECULE WHICH
IS EASILY SOLUBLE AND BOLGTED
AND MADE INTO A AN EASILY PUT
INTO SOLUTION, THE LIPID DOESN'T
LIKE TO BE IN SOLUTION, IT LIKES
TO BE IN A MEMBRANE SO HOW DOES
IT GET RELEASED AND WHO RELEASES
IT AND HOW MUCH IS THERE AND HOW
IS IT TAKEN UP?
INHIBITORY NEURONS ARE ALSO
CONTROLLED BY FAST INHIBITION,
THAT'S ANOTHER 1 OF SIGNIFY HAD
MONITOR CHAMBER LAN'S LOVES, I
DIDN'T COVER THAT TODAY FOR THE
SAKE OF TIME.
THERE'S A CONVERGENCE BETWEEN
THE GENETICS LPIT, AND THE
PHARMACOLOGY OF CANNABINODIs,
BUT ACTUALLY CBD CAME ABOUT
BECAUSE A CHILD HARLOT HAD AND
THE PARENTS MOVED TO COLORADO TO
LOOK INTO THIS AND THAT'S WHAT
SET THE FIRE AND THAT'S WHAT LED
THE FDA TO LOOK INTO THIS AND
FOR THEM TO RUN THE UNBIASED
TRIALS AND WHICH IS BETTER THAN
TOTAL RESULTS, YOU NEED A LARGE
NUMBER OF PATIENT ANDS YOU NEED
TO MAKE IT SOUND FOR THE FDA
APPROVAL BUT THEY MANAGED TO GET
ALL THE WAY FROM 1 PATIENT AND 1
BRAVE FAMILY, ALL THE WAY TO A
DRUG.
AND FINALLY THE IMPORTANCE OF
NONCODING REGIONS, THIS IS THE
FRONTIER OF NEUROSCIENCE AS WELL
AS MANY OTHER FIELDS AND IT
BEHOOVES US TO FIND WAYS OF
TEAMING UP TOGETHER AND STUDYING
THE REGULATION OF THE REGULATORS
BY THE NONCODING REGIONS.
I WOULD LIKE TO THANK YOU VERY
MUCH FOR YOUR ATTENTION.
IT'S A LITTLE STRANGE TO BE HERE
AT THE HEART OF SCIENCE IN THE
U.S. AT LEAST AMONG PEOPLE WHO
ARE DOING SUCH GREAT WORK, I
HAVE A LOT OF RESPECT FOR THE
PIONEERING STUDIES OF PEOPLE
STUDYING INTERNEURONS, STUDYING
CANNABINOID RECEPTORS, STUDYING
SIGNALING IN MANY FORMS,
STUDYING HOW THE BRAIN WORKS AND
IT'S A PLEASURE FOR ME TO BE
HERE AND TALK TO YOU TODAY.
THANK YOU.
KD--SALLY ASH PLAWZ
[ APPLAUSE ]
>> I LITTLE PAST THE HOUR BUT I
WELCOME A QUESTION FROM THE
AUDIENCE.
DR. RAWLING.
>> HI, HAVE YOU LOOKED AT THE
HIGH LEVELS OF FEIGN
ILLEGALSALLA 9OT GPR55 CIRCUIT.
>> NO, WE HAVEN'T.
>> IF YOU HAVE AN E-MAIL MEAS
SEND ME--
>> NO, IT'S JUST A PERSONAL
OBSERVATION FOR SOMEONE WHO USED
TO HAVE FEIGN ILLEGALS ALANINE
INDUCED SEIZURES.
>> REALLY?
, NOT SOMETHING I WOULD HAVE
EXPECTED BUT THANK YOU FOR THAT
INFORMATION.
>> IN YOUR MODEL CIRCUIT, IT
ONLY HAD POSITIVE FEEDBACK, SO,
IN THE LPI CIRCUIT, NOT EVERY
HIPPOCAMPUS IS PRONE TO
SEIZURES, SO WHAT DO YOU THINK
HAPPENS IN THE HEALTHY BRAIN TO
TASM DOWN OTHER ENDOCANNABINODS
IN THE SYSTEM?
>> THERE'S LPA, LIAISONS O
POSFETAL ACID SO IT HAS A
PHOSPHATE GROUP WHERE THE SUGAR
RING IS, SO IT'S LIAISONS O
PHOSPHO TITLE ASIT OLDER PEOPLE,
WITHOUT THE ACITOL, AND REMINDS
ME OF ROBERT DENEUROLOGIST O
ORDERING THE CHICKEN SALAD AND
TELLING THEM TO HOLD EVERYTHING
SO HE BEING GET TOAST.
SO IF YOU HAVE THE MIX OF LPA
AND LPI, YOU COULD HAVE A SYSTEM
TO DIAL YOU UP ANYWHERE YOU WANT
TO BE IN THE COORDINATES, IT'S
LIKE MY GOING TO WORK BY GOING
EAST AND NORTH OR GOING NORTH
AND THEN EAST.
SO THOSE 2 SYSTEMS ARE THE BEAR
MINIMUM THAT'S NEEDED IN ORDER
TO FOR LIPIDS TO DIAL UP
WHATEVER STRENGTH OF E AND I,
YOU WANT.
NOW LPA IS KNOWN TO DOWN
REGULATE BOTH EXCITATORY AND
INHIBITORY TRANSMISSION, BUT IT
HASN'T BEEN STUDIED IN THE SAME
TEDIOUS WAY WE DID IT TO FIGURE
OUT WHAT MAKE ITS REAL RELEASE
AND HOW STRONG IS IT AND IS IT
PART OF A FEEDBACK LOOP, BUT
THAT'S THE NEGATIVE COMPONENT,
SO LPI IS THERE, TO MAKE THE
HIPPOCAMPUS LIVE DANGEROUSLY,
LIVE NEAR THE EDGE, YOU KNOW PUT
YOUR BLIND FOLD ON, PUT YOUR
FOOT ON THE ACCELERATOR, TAKE
OFF THE BRAKES BUT THERE HAS TO
BE A COUNTERACTING TO THAT AND I
THINK IT COULD BE LPA OR IT
COULD BE AN ENDOGENOUS
CANNABINOID ANDACKING IN THE WAY
YOU'RE ALLUDING TO SO THE
COMBINATION OF HOW THESE WORK
TOGETHER IS UNCHARTED TERRITORY.
THANKS FOR THE QUESTION.
IN, THANK YOU EVERYONE FOR
COMING AND PLEASE JOIN US FOR
THE RECEPTION IN THE HALL, IN
THE LOBBY.
GOODBYE.
THANK YOU.
[ APPLAUSE ]