NOKAY, WE ARE NOW 2 MINUTES
OFFICIALLY INTO IT SO I THINK
IT'S TIME TO START.
IT'S MY PLEASURE TO INTRODUCE
BETTY DIAMOND TODAY.
BETTY RECEIVED AN M. D. FROM
HARVARD MEDICAL SCHOOL AND SHE
DID HER RESIDENCY IN INTERNAL
MEDICINE AT COLUMBIA PRES BI
TERRIAN MEDICAL CENTER AND A
POST DOCTORAL FELLOWSHIP IN
IMMUNOLOGY WITH MATTHEW SHARP AT
THE ALBERT EINSTEIN.
NOW BETTY IS THE HEAD OF THE
AUTOIMMUNE DISEASE CENTER AND
DIRECTOR OF THE INSTITUTE FOR
MOLECULAR MEDICINE AT THE
FEINSTEIN INSTITUTE FOR MEDICAL
RESEARCH.
AND HER RESEARCH FOR THOSE
MANYIERS HAS FOCUSED ON THE
INDUCTION AND PATHOGENICITY OF
ANTIDNA ANTIBODIES IN SYSTEMIC
LUPUS HEMAITOSEIS, SLE FOR
SHORT.
HER RESEARCH FOCUSED MAINLY ON
THE ANTIBODIES BUT IS PART OF
THE BROAD AND VERY PROLIFIC
RESEARCH PORTFOLIO THAT SHE HAS
AND HER CV LISTS ABOUT 300
PUBLICATIONS.
SHE SHOWED THAT SOMATIC MUTATION
OF IMMUNOGLOBULIN GENES CAN
GENERATE AUTOIMMUNO ANTIBODIES,
AND PARTICULARLY C1 Q, SHE
STUDIED THE IMPACT OF SEX
HORMONES ON B-CELL SELECTION AND
DEFINED FUNCTIONALITY OF SEVERAL
SLE RISK ALLELES.
HER LAB ALSO DEMONSTRATED THAT A
SUBSET OF THE NTD AND ANTIBODIES
CAN CAUSE NEUROPSYCHIATRIC LUPUS
AND THIS IS GOING TO BE THE
SUBJECT OF TODAY'S TALK.
I DON'T NEED TO SAY THAT SHE
RECEIVED A GAZILLION AWARDS AND
I HAVE A LIST HERE IF I READ ALL
OF THEM IT WILL TAKE PROBABLY
HALF OF OUR TIME.
SO I WILL JUST MENTION THAT HE
HAS BEEN ELECKED TO THE
INSTITUTE OF MEDICINE, IS A PAST
PRESIDENT OF THE AMERICAN
ASSOCIATION OF IMMUNOLOGISTS,
AND JUST A COUPLE OF YEARS AGO,
SHE WAS ELECTED TO THE NATIONAL
ACADEMY OF SCIENCES.
I THINK THAT NEXT IS THE LASKER,
SO WITHOUT FURTHER ADO, BETTY I?
[ APPLAUSE ]
NSO IT'S A PLEASURE TO BE HERE
TODAY.
AND OBVIOUSLY I WILL TALK ABOUT
NEUROPSYCHIATRIC LUPUS, WELL,
IT'S OBVIOUSLY IMPORTANT FOR
PATIENT WHO IS HAVE LUPUS BUT IN
ADDITION THE PARADIGM THAT I'M
GOING TO PRECEPT TO YOU I THINK
IS PROBABLY SOMETHING THAT
EXISTS IN MANY BRAIN DISEASES
AND THEREFORE WHAT WE LEARN IN
TODAYING LUPUS MAY HAVE MUCH
BROADER APPLICATION.
SO THESE ARE DISCLOSURES EMPLOY
I JUST THINK IT'S VERY FUNNY
THAT ALMOST ALL OF THESE ARE CAR
T-CELL COMPANIES.
YOU PROBABLY KNOW THAT CAR
T-CELLS ARE NOW THE NEW PANACEA
FOR LUPUS SO WE'LL SEE.
I THINK EVERYBODY KNOWS THAT
LUPUS IS AN AUTOIMMUNE DEC THAT
PRIMARILY AFFECTS WOMEN.
AND WHAT YOU NEED TO KNOW ABOUT
LUPUS FOR THE PURPOSES OF WHAT I
WILL TALK ABOUT TODAY IS THAT
THERE--I'LL TRY THIS 1--THAT
THERE ARE NEUROPSYCHIATRIC
MANIFESTATIONS OF THE DISEASE
AND THE OTHER THING YOU HAVE TO
KNOW IS THAT ANTIDNA ANTIBODIES
ARE REALLY DIAGNOSTIC OF THE
STEEZ AND THEY CAUSE TISSUE
INJURY IN SKIN AND IN KIDNEY AND
ARE CLEARLY IMPORTANT IN
PATHOGENESIS.
AND INDEED, WITH THE EXCEPTION
OF ACCELERATED ABNORMAL GLUCOSE
TOLERANCELER O SCLEROSIS, MOST
OF WHAT WE KNOW ABOUT TISSUE
INJURY IN LUPUS BEGINS WITH
AUTOANTIBODIES.
SO THESE ARE 19 DIFFERENT
SYNDROMES THAT BE ATTRIBUTED TO
LUPUS, IN THE ABSENCE OF ANY
OTHER REASON FOR HAVING THEM AND
AS YOU CAN SEE SOME AFFECT THE
PERIPHERAL NERVOUS SYSTEM AND
THOSE THAT AFFECT THE CENTRAL
NERVOUS SYSTEM CAN EITHER BE
FOCUS MANIFESTATIONS OF THE
DISEASE WHICH INCLUDE VASCULAR
DISEASE IN THE BRAIN OR DIFFUSE
MANIFESTATIONS OF DISEASE.
THE PREVALENCE OF
NEUROPSYCHIATRIC MANIFESTATIONS
IN LUPUS IS REALLY BEEN VERY,
VERY GREAT, AND IN COHORTS FROM
MANY DIFFERENT PLACES, IT'S
REALLY, YOU KNOW OVER 50% OF
PATIENTS WILL HAVE SOME
MANIFESTATION OF
NEUROPSYCHIATRIC LUPUS, THE ONLY
COHORT IN WHICH THAT'S NOT TRUE
IS A COHORT IN WHICH COGNITIVE
IMPAIRMENT HAD TO BE ACTUAL
DEMENTIA AND WHILE COGNITIVE
IMPAIRMENT AS YOU CAN SEE IS
FREQUENT IN PATIENTS WITH LUPUS,
DEMENTIA IS REALLY QUITE, QUITE
RARE.
COGNITIVE DYSFUNCTION AND MOOD
DISORDERS ARE BOTH NONFOCAL CNS
MANIFESTATIONS OF DEC, ARE 2 OF
THE MOST COMMON MANIFESTATIONS
OF DEC AND IN FACT, PATIENTS
WITH LUPUS WHEN ASKED TO
IDENTIFY THOSE MANIFESTATIONS OF
DEC THAT MOST INTERFERE WITH
QUALITY OF LIFE, HAVE SAID, PAIN
FATIGUE AND COGNITIVE
IMPAIRMENT, SO IT'S A COMMON
MANIFESTATION AND 1 THAT IS
IMPORTANT TO THE PATIENTS
THEMSELVES.
SO HOW DID WE GET INVOLVED IN
STUDYING NEUROPSYCHIATRIC LUPUS
BECAUSE WE NEVER REALLY SET OUT
TO STUDY NEUROPSYCHIATRIC LUPUS.
WHAT WE WERE LOOKING AT WAS THE
STRUCTURAL CORRELATES OF DMA
BINDING, SO WE WILL BUSY
PERFORMING SIGHT DIRECTED MUTE O
GENESIS OF AN ANTIDNA ANTIBODY
THAT WE HAD CLONED FROM A MOUSE
R4A AND LOOKING AT THE BINDING
OF THE ANTIBODY OF THE MUTANTS
TO DNA AND WHAT WE ACTUALLY
FOUND WAS A MUTANT THAT HAD A 10
FOLD HIGHER APPARENT AFFINITY
FOR DNA BUT DID NOT DEPOSIT INTO
KIDNEYS LIKE THE PARENTAL
ANTIBODY DID AND SO WE SAID WELL
THAT ANTIBODY MUST HAVE A
DIFFERENT FINE SPECIFICITY, AND
SO WE WENT TO A PEPTIDE LIBRARY,
A PHAGE DISPLAY LIBRARY AND
LOOKED FOR WHAT THE PARENT R4A
ANTIBODY WOULD BIND AND MUTANT
WOULD BOUND AND INDEED THEY
BOUND DIFFERENT PEPTIDES BUT
WHAT WAS MOST AMAZING IS THAT
THE PARENT ANTIBODY BOUND TO A
CONSENSUS SEQUENCE, PRESENT IN
THE GLUE N2A AND GLUN2 B
SUBUNITS OF THE NMDA VECTOR
EMPLOY SO THAT WAS EXSEATING TO
US AND SUGGESTED WE MIGHT HAVE
THE FIRST ACTUAL MECHANISTIC
CLUE ABOUT NONFOCAL
MANIFESTATIONS OF
NEUROPSYCHIATRIC LUPUS, BUT
BEFORE WE WENT FURTHER, WE
WANTED TO MAKE SURE THAT
ANTIBODIES WITH THIS SPECIFICITY
ACTUALLY EXISTED IN INDIVIDUALS
WHO HAD LUPUS, BECAUSE IF THIS
WAS JUST 1 MOUSE MONOCLONAL THAT
HAD THIS CROSS REACTIVITY, IT
WASN'T GOING BE OF INTEREST TO
US, BUT IF IT WAS A SPECIFICITY
THAT WAS PRESENT IN PATIENTS,
THEN ALL OF A SUDDEN IT BECAME
VERY INTERESTING.
AND WHAT YOU CAN SEE HERE IS
THAT PATIENTS WITH LUPUS HAVE AN
ELEVATED TITERS OF THESE
ANTIBODY ANDS THOSE WITH
NEUROPSYCHIATRIC MANIFESTATIONS,
MORE SO, AND IT'S PRESENT NOT
JUST IN THE SERUM OF PATIENTS
WITH LUPUS, BUT IT'S ALSO
PRESENT IN THE CEREBRAL SPINAL
FLUID, SO THIS IS CEREBRAL
SPINAL FLUID IN RED, PATIENTS
WITH NONFOCAL CNS
MANIFESTATIONS, PNS IS
PERIPHERAL MANIFESTATIONS OF
LUPUS AND S IS A GROUP OF
PATIENT WHO IS WERE UNDERGOING
SURGERY FOR OTHER REASONS AND
PATIENTS WITH LUPUS UNDERGOING
SURGERY FOR OTHER REASONS AND
CONSENTED TO HAVE AN LP DONE AND
YOU CAN SEE THAT IT'S THEREFORE
BOTH IN SERUM AND CSF OR ABOUT
30% OF THE PASHTS.
SO I'M NOT A NEUROSCIENTIST, SO
THIS SLIDE INCLUDES EVERYTHING I
KNOW ABOUT THE NMDA RECEPTOR
WHICH IS THAT IT'S MADE UP OF 2
GL U AND 1 SUBUNITS AND GLUN2
SUBUNITS AND AGONIST IS FLUTE
MATE AND THIS CAUSES CALCIUM
INFLUX INTO THE CELL, IT'S
CRITICAL IN LEARNING AND MEMORY
AND TOO MUCH CALCIUM INFLUX
RESULTS IN WHAT IS CALLED FLUTA
MITTERGIC OR CYTOTOXIC CELL
DEATH EMPLOY I SHOULD ALSO SAY
RIGHT AWAY THAT THIS IS NOT THE
ANTIBODY TO THE NMDA RECEPTOR
THAT HAS BEEN IDENTIFIED IN
PATIENTS WITH LIMBIC
ENCEPHALITIS OR BREAN ON FIRE,
THOSE ANTIBODIES ARE DIRECTED TO
THE GLUN1 SUBUNIT.
THEY CAUSE AN INTERNALIZATION OF
THE RECEPTOR AS SOON AS
ANTIBODIES DISAPPEAR, THE
RECEPTOR IS EXPRESSED ON THE
SURFACE AGAIN AND SYMPTOMS LAST
ONLY AS LONG AS THE ANTIBODY IS
PRESENT AND THEN MOST
INDIVIDUALS RETURN TO NORMAL
BRAIN FUNCHES AND YOU WILL SEE
THAT IS NOT TRUE IN LUPUS.
SO WE WANTED TO DEVELOP A MODEL
WHERE WE COULD STUDY THESE
ANTIBODIES AND I SHOULD SAY THAT
THESE ANTIBODIES, ANTIBODIES
WITH THIS CROSS REACTIVITY
COMPRISE 60% OF THE ANTIDNA
ANTIBODIES, SEEN IN THE MRL,
LPR, MOUSE, WHICH IS A MUCH
STUDIED MODEL FOR LUPUS, AND
ABOUT 60% OF THE ANTIDNA
ANTIBODIES THAT ARE SEEN IN NZBW
MICE, THE ORGT MOST STUDIED
MOUSE MODEL OF LUPUS, BUT THOSE
MODELS HAVE OTHER AUTOANTIBODIES
AND LOTS OF CYTOKINE
PROTURBATIONS, PLUS IF THEY WERE
BORN TO DAMS THAT ALREADY HAD
IMMUNOLOGIC ABNORMALITIES, THEY
COULD HAVE ABNORMAL BRAIN
FUNCTION FROM THE VERY
BEGINNING, BRAIN DEVELOPMENT, SO
WE WANTED TO TAKE
NONSPONTANEOUSLY AUTOIMMUNE MICE
AND MAKE THEM MAKE THIS ANTIBODY
AND THEN BE ABLE TO ASK WHAT IS
THIS ANTIBODY DO.
SO WE IMMUNIZED NORMAL MICE WITH
THE WEPTIDE, MULTIMERRIZED DOWN
A POLYSIGNIFY SEEN BACKBONE OR
IMMUNIZED WITH THE POLYLYSINE
BACKBONE ALONE AND WHAT YOU CAN
SEE IS THAT MICE IMMUNIZED WITH
THE PEPTIDE MAKE ANTIBODIES,
ANTIPEPTIDE O TIDE ANTIBODIES
AND ANTIDNA ANTIBODIES AND IN
THE ANTIDNA ANTIBODIES, ARE AT
THE SAME TITER AS THOSE YOU SEE
IN NCBW MICE AND THEY DEPOSIT IN
KIDNEY SO THEY WERE A GOOD LUPUS
PATHOGENIC ANTIBODY.
AND WE CALL THESE ANTIBODIES DNR
ABs AND YOU WILL SEE THAT
THROUGH THE REST OF THE TALK
EMPLOY BUT WHEN WE WENT TO LOOK
IN THE BRAINS OF THESE MICE,
THEIR BRAINS WERE PERFECTLY
NORMAL AND SO, WE THOUGHT, WELL,
THAT'S BECAUSE THERE'S A BLOOD
BRAIN BARRIER THAT WORKS NOT
JUST TO PROTECT THE BRAIN FROM
IMMUNO GLOBUE LINE BUT ACTUALLY
FCRN HAS AN ABNORMALITIES
LUMINAL LOCALIZATION IN
ENDOTHELIAL CELLS IN THE BRAIN
AND TRANSPORTS IMMUNOGLOBULIN
OUT OF THE BRAIN BUT THERE ARE
MANY INSULTS TO THE INTEGRITY OF
THE BLOOD BRAIN BARRIER AND WE
DECIDED TO USE LPS TO DISRUPT
THE BLOOD BRAIN BARRIER TO MODEL
INFECTION WHICH IS COMMON IN
PEASHTS WITH LUPUS, BY VIRTUE OF
THE IMMUNOSUPPRESSION THERE ON,
AND IT ALSO LPS ALSO GIVES RISE
TO INCREASED TITERS OF MANY OF
THE SAME CYTOKINES, THE
CHARACTERIZE LUPUS FLAIRS.
SO WE IMMUNIZE MICE WITH THE
PEPTIDE OR JUST WITH THE
POLYLYSINE BACKBONE AND THEN WE
GIVE LPS AND THE MICE THAT IMET
THE PEPTIDE IN LPS, ARE DNR AB
POSITIVE MICE AND THE MICE THAT
GET THE POLYLYSINE BACKBONE IN
LPS, ARE DNABRNEGGATIVE MICE.
AND WHAT WE FOUND WERE 2 STAGES
OF PATHOLOGY, 1 IS ACUTE
NEURONAL DEATH, AND YOU SEE THAT
HERE WHERE YOU CAN COUNT THE
PARAMETRAL NEURONS IN THE
HIPPOCAMPUS BECAUSE LPS OPENS
THE BLOOD BRAIN BARRIER
SELECTIVELY IN THE HIPPOCAMPUS,
AND WHAT WE FOUND WAS EXCITE O
TOXIC DEATH BUT THERE'S NO
INFILTRATION OF BLOOD BORN
INFLAMMATORY CELLS, IT'S A BLAND
PATHOLOGY.
AND THEN, NOT AT 2 WEEKS AFTER
GIVING LPS, BUT STARTING AROUND
4-6 WEEKS, AND CLEARLY PRESENTED
8 WEEKS AFTER LPS, WE SEE A LOSS
OF DENDRITIC HARBORRIZATION IN
THE REMAINING NEURONS AND WHAT
YOU SEE ON THE--OVER HERE IS
THAT BY 2 WEEKS AFTER LPS,
THERE'S NO MORE ANTIBODY IN THE
BRAIN.
SO THIS IS OCCURRING, THIS
DAMAGE IS ACUING CUING--ACCRUINN
THE ABSENCE OF ANTIBODIES STILL
BEING IN THE BRAIN.
AND WHAT WE ALSO FOUND WAS
MICROGLIAL ACTIVATION AND A
QUIESCENT MICROGLIA IS A LACY
LOVELY STRUCTURE AND AN
ACTIVATED MICROGLIA IS A MUCH
MORE ROBUST TRUCTURE WITH
EXPRESSION OF CD69 AND BETH
STEPHENS LAB HAS WORKED OUT A
CORING SYSTEM FOR MICROTBLEEL
ACTIVATION AND YOU START SEEING
THEY BECOME ACTIVATED 2 WEEKS
BUT BY 8 WEEKS THEY ARE VERY
ACTIVATED EMPLOY AND THIS IS A
BET SCAN USING PBR 28 WHICH WILL
BIND TO ACTIVATED GLIAL CELLS
BOTH MICROGLIA AND DENDRITIC
CELLROCYTES, BUT WHAT YOU CAN
SEE IS GLIAL CELLS IN THE
HIPPOCAMPUS OF THE MICE THAT
HAVE THE LUPUS AND BRAIN, AND WE
FOCUSED OUR ATTENTION TO SPATIAL
MEMORY IN THESE MICE AND WHAT
YOU SEE HERE IS AN OBLIGATIONS
YECT MEMORY TEST WHERE YOU PUT A
MOUSE IN A CHAMBER WITH 2
OBJECTS, YOU REPLACE THE MOUTH
WITH THE DIFFERENT OBJECT, PUT
IT BACK IN A NORMAL MOUSE WILL
SPEND MOST TIME STUDYING THE
NOVEL OBLIGATIONS YECT AND
INDEED, THESE MICE WITH THE
LUPUS LIKE ANTIBODY PENETRATING
THE BRAIN HAVE NORMAL OBJECT
MEMORY, TEST IS YOU PUT THE
MOUSE IN THE CHAMBER WITH 2
OBLIGATIONS JEKS, TAKE THE MOUSE
OUT AND A NORMAL MOUSE WILL
SPEND NORMAL TIME IN THE CHAMBER
EXPLORING THE OBJECT IN THE NEW
LOCATION, BUT WHAT YOU SEE HERE
IS THAT THE LUPUS MICE DID NOT
AND THIS FOCUS ON SPATIAL MEMORY
WAS SUGGESTED TO US BY A PATIENT
WHO CAME TO CLINIC 1 DAY AND
SAID IT WAS THE WE'RED THING,
BUT WHEN SHE GOT OFF THE BUS TO
GO HOME, SHE COULDN'T REMEMBER
BUT THIS EPITOMIZED THAT
PROBLEM.
SO HIPPOCAMPAL CELLS ARE THE
CELLS THAT ARE RESPONSIBLE FOR
THE SPATIAL MAPS WE MAKE,
THEY'RE CALLED PLACE CELLS AND
THEY FIRE IN A PARTICULAR
LOCATION.
EACH NEURON WILL FIRE IN A
PARTICULAR LOCATION AND THE SAME
LOCATION EVERY TIME.
AND THE AREA OVER WHICH THAT
NEURON WILL FIRE IS CALLED THE
PLACE FIELD.
AND YOU CAN SEE THAT BEFORE LPS
AND AFTER LPS, THE PLACE FIELD
SIZE IS THE SAME IN THE
PENETRATION OF THOSE WHAT HAD
NONLUPUS SERIES POINTSOLOGY IN
THEIR BRAIN BUT IN THE MICE THAT
HAD THE LUPUS LIKE ANTIBODY BEN
TRAIT THE BRAIN, THE--EACH
NEURON HAS A BIGGER PLACE FIELD.
SO FIRES OVER A WIDER AREA AND
TO ME IT'S THAT PLACE CELL
FIRING ON 2 BLOCKS INSTEAD OF
JUST ON 1 BLOCK.
AND SO WHAT WE LEARNED IS THAT
MICROGLIA ARE REQUIRED FOR THE
LOSS OF DENDRITICAR BORRIZATION
IN THE RESIDUAL NEURONS AND C1 Q
IS REQUIRED FOR THE LOSS OF
ARBORIZATION ON THE RESIDUAL
YOU'REONS IN THE DNA AB POSITIVE
MICE AND THIS SUGGIESTED TO US
THAT WHAT WE WERE SEEING WAS
ACTUAL PRUNING.
SO THAT THE SYNAPSES WERE BEING
EATEN BY THE ACTIVATED MICROGLIA
AND WHEN THE SYNAPSES ARE EATEN
OFF A DENDRITE, THERE'S
RETRACTION OF THAT DENDRITE AND
HENCE DECREASED DENDRITIC
ARBORIZATION.
AND INDEED, YOU CAN SEE C1 Q
SITTING ON THE SYNAPSES, SO WE
ASKED HOW DOES C1 Q GET TO THE
SYNAPSES.
WHAT TARGETS IT THERE AND 1 OF
THE OBSERVATIONS THAT WE MADE
WAS THAT IN THE MICE THAT HAD
THE LUPUS, ANTIBODY, PENETRATE
THEIR BRAIN, THERE WAS EVIDENCE
FOR SECRETION OF HMGB1.
SO IT WAS KNOWN FROM OTHERS THAT
AS YOU ACTIVATE THE NMDA
RECEPTOR, IN A DOSE DEPENDENT
FASHION, WELL IS SECRETION OF
HMGB1 WHICH IS NORMALLY A
NUCLEAR PROTEIN INVOLVED IN
PROAMA TIN STRUCTURE BUT IS
ACTIVELY ACETALATED AND SECRETED
BY STRESS CELLS, AND ACTIVATED
CELLS AND AS I SAID HAS BEEN
KNOWN TO BE SECRETING NEURONS TO
THE NMDA RECEPTOR.
SO OUR HYPOTHESIS WAS THAT HMGB1
WHICH HAD ALSO PREVIOUSLY TO OUR
WORK BEEN SHOWN TO BIND TO
GASHING WELL, UN2 SUBUNITS OF
THE NMDA RECEPTOR ACTUALLY
SERVED AS A BRIMMING TO BRING C1
Q TO THIS SYNAPSE BECAUSE IN OUR
STUDIES OF C1 Q, SOME OF YOU MAY
KNOW THAT C1 Q IS THE BIGGEST
RISK FACTOR FOR LUPUS AND OVER
90% OF INDIVIDUALS WHO HAVE C1 Q
DEFICIENCY HAVE LUPUS SO WE
WERE--ARE INTERESTED IN HOW C1 Q
MAINTAINS'MUNE HOMEOSTASIS.
BUT WE HAD SHOWN THAT C1 Q BINDS
HMGB 1.
SO WHAT YOU SLEEP APNEA SLEEP D
OBESITY--WHAT YOU SEE HERE IS IN
FACT THAT HMGB1 WILL BIND TO
SYNAPSES AND THAT C1 Q WILL BIND
TO THE HMGB 1 ON SYNAPSES.
SO, INDEED, THERE IS A BRIDGE
FORMED BY THE SECRETED HMGB1
THAT BRINGS C1 Q TO THE SYNAPSE
TO ALLOW THE SYNAPSE TO BE EATEN
BY ACTIVATED MICROGLIAL CELLS.
SO 1 OF THE QUESTIONS WE THEN
ASKED IS WHETHER THIS HMGB1 HAS
TO COME FROM NEURONS.
AND SO WE ACTUALLY DELETED HMGB1
IN NEURONS AND WHAT YOU
CAN--WHAT WE FIND IS THAT THESE
MICE HAVE THE SAME INITIAL LOSS
OF PARAMETRAL NEURONS BUT
THERE'S NO MICROGLIAL
ACTIVATION.
AND WHEN YOU DO THIS SAME STUDY,
WHERE YOU DELETE THE HMGB1 FROM
MICROGLIA, YOU DON'T HAVE THIS
PHENOTYPE.
THEY DEVELOP DISEASE JUST LIKE
THE WILD-TYPE MICE DO.
SO WHY YOU NEED THE HMGB1 TO BE
SECRETED BY NEURONS AND IF IT'S
REALLY JUST A MATTER OF
PROXIMITY TO BE ABLE TO GET THE
HMGB1 TO THE SYNAPSE, I'M NOT
SURE.
SO, HAVING SHOWN THAT HMGB1 IS
CRITICAL FOR SYNAPTIC PRUNING,
WE ALSO ASKED WHETHER HMGB1 IS
CRITICAL TO THE MICROGLIAL
ACTIVATION.
BECAUSE EXTRA CELLULAR HMGB1 IS
A DAM, A DAMP ASSOCIATED PAT
EXPERN IT BINDS TO RAGE AND
TLR4.
SO WE THOUGHT THAT THIS SECRETED
HMGB1 MIGHT HAVE THE FUNCTION OF
ALSO ACTIVATING THE MICROGLIA TO
MOVE THEM TO THIS STATE WHERE
THEY ENGAGE IN PRUNING AND WHAT
YOU CAN SEE HERE IS IF YOU
INCUBATE HMGB1 ISOLATED FROM THE
BRAIN, EITHER WILD-TYPE OR RAGE
DEFICIENT MICROGLIA, YOU CAN SEE
THERE'S MUCH LESS ACTIVATION OF
THE DEFICIENT MICROGLIA IN TERMS
OF TNF SECRETION AND PRODUCTION
IN TERMS OF INTERFERON, TYPE 1
PRODUCTION AND SECRETION IL1, ON
THE RIGHT HAND SIDE, YOU SEE THE
RESULTS OF THE INTERFERON
ACTIVATION IN INCREASED
INTERFERON STIMMULED GENES AND
ALSO INCREASED C1 Q.
SO WE DID THE EXPERIMENT TO LOOK
AT THIS MODEL IN MICE THAT
LACKED RAGE.
SO, WE ACTUALLY WERE NOT ABLE TO
MAKE MICE THAT LACKED--THAT WE
WERE ABLE TO--RAGE DEFICIENT
MICE THAT WE WERE ACTUALLY ABLE
TO DEVELOP THE IMMUNIZATION
MODEL IN, BECAUSE RAGE IS ON THE
SAME CHROMOSOME AS MHC, AND THE
MICE HAVE TO BE H2 D IN ORDER TO
MOUNT AN IMMUNE RESPONSE TO THE
PEPTIDE, AND WE WERE NOT ABLE TO
GET A RECOMBINATION EVENT
BETWEEN THE RAGE AND THE MHC.
SO WE USED A MONOCLONAL
ANTIBODY, EITHER A CONTROL
ANTIBODY, THAT DID NOT, CROSS
REACT WITH DNA AND THE NMDA
RECEPTOR, OR AN ANTIBODY THAT
CROSS REACTS WITH THE NMDA
RECEPTOR AND DNA.
AND WHAT YOU CAN SEE, IS THAT IN
THE RAGE KNOCKOUT MICE, WE DID
NOT HAVE MICROGLIAL ACTIVATION
WHEN WE USED THE LUPUS LIKE
ANTIBODY, IT WAS NO DIFFERENT
THAN WHEN WE GAME THE MOUSE THE
NONLUPUS LIKE ANTIBODY AND THEN
LPS, AND IN THE RAGE KNOCKOUT
MICE WE DID NOT GET A LOSS OF
DENDRITIC ARBORIZATION, SO
CLEARLY THE HMGB1 CAN ACTIVATE
MICROGLIA THROUGH BINDING TO
RAGE.
SO OUR INTEREST IS ALWAYS IN THE
PATIENT.
SO WE WERE WONDERING IF THE NEED
FOR MICROGLIAL ACTIVATION GAVE
US A CLUE AS TO A THERAPEUTIC
INTERVENTION.
AND ANGIO TENSEIN ENZYME
INHIBITORS CAN BLOCK ACTIVATION
OF MYELOID CELLS IN THE
PERIPHERY.
THEY ARE USED ALL THE TIME IN
INDIVIDUALS WITH LUPUS BECAUSE
THEY'RE USED TO MANAGE THE RENAL
DEC AND HYPERTENSION, THEY'RE
SAFE, THEY'RE RELATIVELY
INEXPENSIVE COMPARED TO MOST
DRUGS WE USE IN LUPUS, THEY HAVE
BEEN SHOWN TO HAVE SOME BENEFIT
IN ALZHEIMER'S DISEASE, WHICH IS
ALSO CHARACTERIZED BY
MICROTBLEEL PRUNING OF NEURONS,
AND SO, WE DECIDED THAT WE WERE
GOING TO STUDY ACE INHIBITORS.
AND 1 OF THE OTHER OBSERVATIONS
WE MADE IS NEURONS ACTUALLY
PRODUCE ACE ANGIO TENSEIN
CONVERTING ENZYME AND THERE WAS
LOTS MORE ACE IN THE
DNR AB POSITIVE MICE THAN THE
NEGATIVE MICE, AND THERE'S A
RISK ALLELE FOR LUPUS THAT LEADS
TO HIGH PRODUCTION OF ACE.
THE OTHER THING THAT'S IMPORTANT
TO KNOW IS THAT INTERFERON WHICH
IS PRODUCED BY THESE ACTIVATED
MICROTBLEEL CELLS, LEADS TO
INCREASED PRODUCTION OF THE
RECEPTOR 1 FOR ANGIO TENSEIN 2
WHICH TRIGGERS PRO INFLAMMATORY
PATHWAY.
SO WE DECIDED TO USE ACE
INHIBITORS IN BOTH THE
PREVENTION AND THE TREATMENT
MODEL AND CAPP A TRIL IS AN ACE
INHIBITOR THAT CROSSES THE BLOOD
BRAIN BARRIER AND NOW
[INDISCERNIBLE] IS AN INHIBITOR
THAT DOES NOT CROSS THE BLOOD
BRAIN BARRIER AND WHAT YOU CAN
SEE IS THAT IN THE SCORE BELOW
IS THAT CAPTOPRIL LED TO A
DECREASE IN MICROGLIAL
ACTIVATION IN BOTH THE
PREVENTION TRIAL AND ENALAPRIL
DID NOT.
AND IN FACT, CAPTOPRIL RESTORED
NORMAL DENDRITIC NORMALIZE IN
THE IN THE NEURONS AND ENALAPRIL
DID NOT IN BOTH THE PREVENTION
AND TREATMENT MODEL AND IN
THE--IN BOTH MODELS, SPATIAL
MEMORY WAS EITHER PROTECTED OR
RESTORED.
SO THIS WAS A VERY EFFECTIVE
TREATMENT.
SO HOW DOES IT WORK?
WELL WE THOUGHT THAT IT MIGHT
WORK THROUGH UPREGULATION OF
LAYER, IN OUR STUDIES OF C1 Q,
WE IDENTIFIED LAYER 1 AS AN
INHIBITORY RECEPTOR FOR C1 Q.
AND IT WILL BLOCK ENGAGING LAYER
1 WILL BLOCK MONOCYTE ACTIVATION
IN THE PERIPHERY.
AND SO WHAT OUR MODEL WAS WAS
THAT A QUIESCENT MICROGLIA HAS A
CERTAIN NUMBER OF SCAVENGER
RECEPTORS AND A CERTAIN NUMBER
OF LAYER 1 MOLECULES.
AND THEY'RE IN SOME KIND OF
BALANCE AND THE ACTIVATED
MICRODPLIA, WE THOUGHT HAD MORE
SCAVENGER RECEPTORS, FEWER LAYER
1 MOLECULES, SO C1 Q WILL
PREFERENTIALLY BIND THE
SCAVENGER RECEPTORS LEADING TO
PRUNING, AND WHEN WE USE THE ACE
INHIBITOR, THERE IS REEXPRESSION
OF MORE LAYER 1, AND SO NOW,
THERE'S NO LONGER PREFERENTIAL
ACTIVATION OF THE SCAVENGER
RECEPTORS AND SO 1 OF THE
PREDICTIONS, WELL I SHOULD FIRST
SAY THAT IF WE LOOK AT LAYER 1
EXPRESSION ON MICROGLIA,
ISOLATED FROM THE DNRAB POSITIVE
MICE, THERE'S LESS LAYER 1 THAN
FROM DNRA B-NEGATIVE MICE AND IF
YOU LOOK AT THE ENALOPRIL MICE
VERSUS CAPTOPRIL TREATMENT MICE,
THERE'S HIGHER LAYER OF
EXPRESSION LAYER 1 IN THE
CAPTOPRIL, QUIESCENT NOW
QUIESCENT MICRO FLI COLSISSA.
AND 1 OF THE PREDICTIONS IS THAT
ACE INHIBITION WILL NOT BE OF
ANY BENEFIT IN LAYER 1 DEFICIENT
MICE AND INDEED THAT'S THE CASE.
SO THERE'S NO DECREASE IN
MICROTBLEEL ACTIVATION WITH ACE
INHIBITION IN THE LAIR 1
DEFICIENT MICE AND THE LAIR 1
DEFICIENT MICE DO NOT HAVE THEIR
DENDRITIC ARBORIZATION RESTORED
TO NORMAL IN THE PRESENCE OF AN
ACE INHIBITOR.
SO WHAT REGULATES LAYER 1 IN THE
MICROGLIA.
SO I'M SURE THERE ARE MANY, MANY
FACTORS THAT DO THAT, BUT WHAT
WE FOUND IS THAT MICROGLIA
TREATED WITH HMGB1 MAKE
INCREASED IL10 AND IL10 CAN
DECREASE LAYER 1 EXPRESSION IN
MICROGLIA, SO THAT'S AT LEAST 1
MECHANISM.
SO HAVING FOLLOWED SORT OF
CANDIDATE YEENS THAT CAME FROM
OUR OWN EXPERIMENTS OR OUR
READING LITERATURE, WE DECIDED
TO TAKE A MORE UNBIASED APPROACH
AND LIKE EVERYBODY ELSE WE DID
RNASEQ SINGLE CELL RNASEQ ON
MICROGLIA FROM MICE THAT WERE
DNR ABNEGGATIVE, MICE THAT WERE
DNRAB+, AND DNRAB+ MICE TREATED
WITH CAPTOPRIL, AND THE FIRST
THING YOU CAN SEE IS THERE
THERE'S 2 CLUSTERS OF MICROGLIA
AND 1 THAT IS HOMEOSTATIC AND 1
THAT IS REACTIVE.
AND WHAT YOU CAN ALSO SEE IS THE
FREQUENCY OF THE REACTIVE CELLS
IS INCREASED IN THE DNETWORK R
AB-POSITIVE MICE AND DECREASED
BY TREATMENT WITH CAPTOPRIL.
THIS IS A SET OF VOLCANO PLOTS,
AND WHAT YOU CAN SEE IS IN THE
HOMEOSTATIC CLUSTER, THERE'S NOT
ALL THAT MUCH DIFFERENCE IN THE
DNR AB POSITIVE MICE AND
DNR ABNEGGATIVE MICE AND THERE'S
NOT THAT MUCH DIFFERENCE INDUCED
BY CAPTOPRIL TREATMENT.
BUT IN THE REACTIVE POPULATION
WHAT YOU CAN SEE IS THAT THERE'S
A BIG DIFFERENCE BETWEEN DNR
AB-+ AND NEGATIVE, AND ALMOST NO
DIFFERENCE BETWEEN DNR
AB NEGATIVE AND THE CAP TO PRIL
TREATED DNR AB + MICROGLIA.
AND WHEN WE LOOKEDDA THE OUR
CANDIDATE GENES WITHIN THE
SINGLE CELL RNASEQ DATA, IN THIS
REACTIVE POPULATION, WHAT WE SEE
IS THE SAME TRENDS THAT WE
SHOWED WITH HMGB1 INCREASING
EXPRESSION OF INFLAMMATORY
CYTOKINES AND INTERFERON, AND
CAPTOPRIL RESTORING EXPRESSION
MORE TOWARDS NORMAL.
AND I SHOULD JUST SAY THAT ACE
INHIBITORS CAN WORK BY
PREVENTING THE PRODUCTION OF
ANGIO TENSEIN 2 WHICH I TOLD YOU
CAN BIEN TO A PRO INFLAMMATORY
RECEPTOR, AT1 R, BUT THEY CAN
ALSO WORK BY DECREASING THE
DEGRADATION OF BETA KINEIN AND
IT HAS BEEN SHOWN TO SUPPRESS
MONOCYTE ACTIVATION.
SO WE WERE INTERESTED IN WHETHER
THE ACE INHIBITION WAS WORKING
BY PREVENTING THE ACTIVATION OF
THE ANGIO TENSEIN 2 RECEPTOR, OR
WAS WORKING BY INCREASING THE
AMOUNT OF BRAIDA KINEIN PRESENCE
AND WHAT CAN YOU SEE HERE IS
THAT IT CLEARLY CAN WORK BY
DECREASING THE ENGAGEMENT OF THE
ANGIO TENSEIN RECEPTOR BECAUSE
[INDISCERNIBLE] IS AN ANGIO
TENSEIN RECEPTOR BLOCKER AND
[INDISCERNIBLE] THAT'S USED
CLINICALLY IN MANY SITUATIONS
AND WHAT YOU CAN SEE IS THAT
THAT WORKED LIKE CAPTOPRIL TO
RESTORE A NORMAL MICROGLIAL
NEURONAL HOMEOSTASIS IN THE
BLOOD AND THE BRAIN.
SO WHAT'S OUR MODEL, WELL, OUR
MODEL IS THAT THE FIRST STAGE IS
CAUSED BY EXCITE O TOXICITY.
THAT THERE IS LOTS OF HMGB1 THAT
IS BEING SECRETED BY NEURONS
THAT ACTUALLY DON'T DIE BUT GET
ENORMOUS NMDA RECEPTOR
ACTIVATION BY THE PRESENCE OF
THE ANTIBODY.
THAT THAT BOTH TARGETS, HELPS
TARGET SYNAPSES FOR PRUNING AND
ACTIVATES THE MICROGLIA TO
ENGAGE IN PRUNING AND WHEN ACE
INHIBITORS OR ARBs COME ALONG
WE INCREASE LAIR 1 EXPRESSION,
SO THE C1 Q CAN PREFERENTIALLY
BIND TO LAYER 1 AS OPPOSE TO
SCAVENGER RECEPTORS AND WE END
UP WITH MICROFLEEL QUIESCENCE
AND RECONSTITUTION OF THE
DENDRITIC PROCESSES IN THE
NEURONS.
WHAT WAS REALLY MOST SURPRISING
TO US AND I'M GOING TO GET BACK
TO THIS, IS THAT THIS PATHOLOGY
IS SUSTAINED AS FAR AS WE CAN
TELL INDEFINITELY AFTER THE
INITIAL ADMINITRATION OF LPS, SO
12 MONTHS LATER, THERE IS NO
RESOLUTION OF THE INFLAMMATION
IN THE BRAIN.
SO HOW DOES THIS RELATE TO
PATIENTS.
SO THIS IS LOOKING AT BLOOD
BRAIN BARRIER INTEGRITY IN
PATIENTS WITH LUPUS.
AND THESE ARE PATIENTS WHO HAVE
ABNORMAL PERFORMANCE ON
NEUROCOGNITIVE EXAMS, BUT NO
ACUTE CNS DISEASE.
AND WHAT YOU CAN SEE IS THERE IS
SELECTIVE IMPAIRMENT OF BLOOD
BRAIN BIRRIER SURFACESSIER
INTEGRITY IN THE HYPOCAMPUS.
IF YOU TAKE LUPUS SERUM THAT HAS
THESE ANTIBODIES IN IT, AND GIVE
IT INTRAVENOUSLY, THE
IMMUNOGLOBULIN TO A MOUSE OR
GIVE NORMAL SERUM TO A MOUSE OR
FRANKLY GIVE THIS LUPUS IMMUNO
DPLOBUE LYNN DEPLETED OF THIS
ANTIBODY, ON A PEPTIDE AFFINITY
COLUMN TO A MOUSE, AND THEN GIVE
LPS, AND TEST FOR SPATIAL MEMORY
IN A PADDLING MAZE, THE MICE
THAT HAVE THE LUPUS ANTIBODY
PENETRATE THE BRAIN AND BIND THE
HIPPOCAMPAL NEURONS HAVE
IMPAIRED MEMORY FUNCTION.
IF YOU LOOK AT LUPUS PATIENTS
AND YOU ASK WHETHER THEY HAVE
NORMAL OBJECT MEMORY AND NORM AT
SPATIAL MEMORY, WHAT YOU CAN
SEE, IS THAT OUR TEST FOR--WELL,
I SHOULD FIRST SAY, THAT MOST
NEUROPSYCHIATRIC BATTERIES THAT
ARE USED IN THE CLINIC AND THAT
ARE RECOMMENDED FOR USE WITH
PATIENTS WITH LUPUS, DO NOT
DISTINGUISH BETWEEN OBLIGATIONS
YECT MEMORY, AND SPATIAL MEMORY.
SO WE DEVICED A VERY SIMPLE TEST
OR USED A VERY SIMPLE TEST WHERE
THERE ARE 4 OBLIGATIONS YECTS ON
A CARD AND AN OBJECT MEMORY
TEST, IS IF YOU ASK IF THERE WAS
A DOG ON THE CARD, AND THEY SAY
YES OR NO, AND A SPATIAL MEMORY
TEST IS WHETHER THE BANANA WAS
NEXT TO THE LION, AND THEY SAY,
YES OR NO.
AND WHAT YOU CAN SEE, HC IS
HEALTHY CONTROLS, DNRB
ABNEGGATIVE AND PATIENTS WITH
LUPUS WHO DO NOT HAVE THIS
ANTIBODY, AND DNRAB+ IS PATIENTS
WITH LUPUS WHO DO HAVE THIS
ANTIBODY AND WHAT YOU CAN SEE IS
THAT THEY ALL PERFORM
SIMILARLY ON AN OBLIGATIONS YECT
RECOGNITION TASK BUT ON A
SPATIAL MEMORY TEST, THOSE WHO
HAVE THE--THOSE PATIENTS WITH
LUPUS WHO HAVE THIS ANTIBODY,
HAVE AN IMPAIRMENT IN THEIR
SPATIAL MEMORY.
SO THE PATIENT WHO CAME TO US IN
CLINIC AND MADE US THINK ABOUT
SPATIAL MEMORY, WAS ACTUALLY
ENORMOUSLY INSTRUCTIVE.
IF YOU PERFORM FDG PET SCANS
ON PATIENTS WITH TO LOOK WITH
LUPUS TO LOOK AT TBLUICOSE
UPTAKE, WHAT YOU SEE IS
INCREASED GLUCOSE UPTAKE IN THE
PIP O CAMPUS THAT CORRELATES
WITH MEMORY DYSFUNCTION AND IF
THEY HAVE THE ANTIBODY AS WELL
THERE'S EVEN A GREATER
CORRELATION WITH MEMORY
DYSFUNCTION.
AND THEN IF YOU LOOK AT
FRACTIONAL AND ISOTROPY WHICH IS
REALLY LOOKING AT
MICROSTRUCTURAL INTEGRITY IN THE
BRAIN, WHAT YOU CAN SEE IS THAT
THE GOOD MICROSTRUCTURAL
INTEGRITY, CORRELATES WITH GOOD
PERFORMANCE ON SPATIAL MEMORY,
AND THE GOOD MICROSTRUCTURAL
INTEGRITY CORRELATES WITH NOT
HAVING THE ANTIBODY PRESENT IN
THE SERUM.
SO THOSE WITH POOR
MICROSTRUCTURAL INTEGRITY.
ARE POSITIVE FOR THIS CROSS
REACTIVE ASPECT BODY AND HAVE
IMPAIRED SPATIAL MEMORY AND THIS
IS JUST LOOKING AT THE
MICROSTRUCTURAL INTEGRITY AND
YOU CAN SEE, ARK ROUND THE
HIPPOCAMPUS, THAT THERE'S
REDUCED STRUCTURAL INTEGRITY.
IF YOU LOOK AT PATIENTS FOR
GLIAL CELL ACTIVATION USING THE
SAME PET LIGAND AS WE USED IN
MICE, CAN YOU SEE THAT PATIENTS
WITH LUPUS HAD INCREASED
MICROTBLEEL ACTIVATION IN THE
HIPPOCAMPUS AND I SHOULD JUST
SAY THAT THE BLOOD BRAIN BARRIER
CAN BE IMPAIRED, THE INTEGRITY
OF THE BLOOD BRAIN BARRIER CAN
BE IMPAIRED BY SOLUBLE FACTORS
THAT ARE IN THE CIRCULATION BUT
IT CAN ALSO BE IMPAIRED BY
SOLUBLE FACTORS SECRETED BY
MICROGLIA WITHIN THE CENTRAL
NERVOUS SYSTEM.
AND WHAT YOU SEE, WHEN YOU
FOLLOW PATIENTS OVER TIME, AND
SO, WE'VE DONE LONGITUDINAL
KAN--KANAS AND ASSESSMENTS OF
PATIENTS WITH COGNITIVE
IMPAIRMENT AND LUPUS, AND WHAT
YOU CAN SEE IS THAT THOSE WHO
HAVE BEEN GIVEN AN ACE
ESSENTIALLY ACTING ACE
INHIBITOR, CAPTOPRIL IS NOT THE
1 USED CLINICALLY SO MUCH
ANYMORE, BUT ESSENTIALLYACKING
ACE INHIBITOR HAVE A DECREASE IN
THE HYPER METABOLISM AND THOSE
WHO DID NOT GET CENTRALLY ACTING
ACE INHIBITOR HAVE NO ACE
INHIBITOR OR ACE INHIBITOR THAT
DOESN'T CROSS THE BLOOD BRAIN
BARRIER HAD IN FACT AN INCREASE
IN HYPER METABOLISM IN THE
BRAIN.
SO THAT HAS LED TO A CLINICAL
TRIAL THAT IS ONGOING, TO LOOK
AT A CENTRALLY ACTING, VERSUS A
NONCENTRALLY ACTING ACE
INHIBITOR IN PEOPLE WITH LUPUS,
WHO HAVE INCREASED METABOLISM IN
THEIR HIPPOCAMPUS, ABNORMAL
MEMORY FUNCTION ON
NEUROPSYCHIATRIC TESTING, AND
IT'S REALLY THE CLINICAL TRIAL
MADE IN HEAVEN BECAUSE EVERYBODY
GETS AN ACE INHIBITOR AND SO
EVERYBODY GETS THE BENEFITS OF
THE ACE INHIBITOR FOR BLOOD
PRESSURE CONTROL AND RENAL
PROTECTION, BUT SOME GET AN ACE
INHIBITOR THAT WILL ACTUALLY WE
HOPE SEPRESS MICROFLEEL
ACTIVATION AND SOME GET AN ACE
INHIBITOR THAT WILL NOT
PENETRATE THE BRAIN.
OUR READ OUT FOR THIS CLINICAL
TRIAL IS NEUROIMAGING, BECAUSE
NEUROIMAGING IS BASICALLY
INDEPENDENT OF TIME OF DAY,
INDEPENDENT OF MOOD, AND
NEUROPSYCHIATRIC TESTING.
YOU WOULD HAVE TO HAVE INFINITLY
MORE PATIENTS TO DO THIS KIND OF
STUDY AND USE NEUROPSYCHIATRIC
TESTING AS YOUR END POINT AND WE
FINISHED ENROLLMENT THIS MONTH
AND WE WILL FIND OUT.
SO WE'RE ROWBSLY DOING COGNITIVE
TESTING BUT THAT'S NOT A PRIMARY
END POINT.
ONE OF OUR OTHER END POINTS IS
MICRODPLEEL ACTIVATION.
AND THIS IS WHERE WE COME BACK
TO THE FACT THAT IN MICE THERE'S
NO SPONTANEOUS RESOLUTION OF
THIS INFLAMMATORY PATHOLOGY OVER
TIME SO IN THE MICE, THERE IS 1
TRIGGERING EPISODE AND THEN
THERE IS A POGHT O LOGIC
HOMEOSTASIS THAT STAYS AND STAYS
AND STAYS.
SO WE'VE BEEN STUDYING PATIENTS
WHO WERE IN LONG-TERM REMISSION,
MEANING THEY'VE BEEN IN CLINICAL
REMISSION FOR 3 YEARS OR MORE,
ON NO DRUGS, EXCEPT MAYBE
HYDROXY CHLOR O QUIN, MAYBE NOT,
WHO ALL HAD VERY ACTIVE DEC AT
SOME POINT.
SO THEY HAD ALL PREVIOUSLY
REQUIRED IMMUNOSUPPRESSIVE
THERAPY AND AT LEAST
20-MILLIGRAMS OF COTTERRIC O
STEROIDS BUT THEY'RE NOW ALL
CLINICALLY QUIESCENT.
WE A THEY'RE IN REMISSION,
THERE'S NO EVIDENCE OF
PERIPHERAL INFLAMMATION,
ANYMORE.
AND WHAT YOU CAN SEE, THAT THEY
STILL HAVE INCREASED FDGPET,
INCREASED METABOLISM, GLUCOSE
UPTAKE IN THE HIPPOCAMPUS, THE
PATIENTS IN REMISSION OR IN
BLUE, THE PATES WHO ARE--WHO
STILL HAVE ACTIVE DISEASE AND
ARE ON IMMUNOSUPPRESSIVE THERAPY
ARE IN GREEN.
THEY STILL HAVE BLOOD BRAIN
BARRIER ABNORMALITIES, AND THEY
TILL HAVE EVIDENCE FOR
MICRODPLEEL ACTIVATION.
SO, WE THINK THAT THIS IS A
NONRESOLVING INFLAMMATION IN THE
BRAIN AS WELL WHILE THE NO
INFLAMMATION IS NO LONGER
DETECTABLE, THE BRAIN
INFLAMMATION IS STILL THERE AND
OBVIOUSLY WE THINK THERE ARE BIG
CLINICAL IMPLICATIONS OF THIS
OBSERVATION.
SO WHAT ARE OUR CONCLUSIONS?
WE THINK THESE ANTIBODIES
CONTRIBUTE TO COGNITIVE
IMPAIRMENT IN LUPUS, BUT I DON'T
FOR A MOMENT, WANT YOU TO THINK
THIS IS THE ONLY MECHANISM, THE
ONLY TRIGGER FOR COGNITIVE
IMPAIRMENT IN LUPUS, THERE ARE
OTHER ANTIBRAIN ANTIBODIES.
A GROUP IN CHILE HAS SHOWN THAT
THERE ARE GROUP OF ASPECT BODIES
THAT ARE HONE IN A SMALLER
SUBSET OF PATIENTS AND DNA
ANTIBODIES THAT THOSE CROSS
REACT WITH A CELL SURFACE
MOLECULE ON NEURONS THAT
REGULATE AMPORECEPTOR
TRAFFICKING AND THAT THE
PENETRATION OF THOSE ANTIBODIES
INTO THE BRAIN WILL ALSO CAUSE A
MEMORY IMPAIRMENT AND A
COGNITIVE PHENOTYPE AND I'M SURE
THERE ARE MANY OTHERS.
THAT MICROTBLEEL ACTIVATION WITH
INCREASED C1 Q SECRETION WE
THINK IS A KEY COMPONENT OF THE
COGNITIVE IMPAIRMENT BECAUSE WE
THINK THAT THE NEURONAL HMGB1
WHICH CAN LEAD TO INCREASED C1
Q, SECRETION BY MICROFLEEL CELS
AND THE C1 Q LEAD TO THIS
MALADAPTIVE HOMEOSTASIS WHICH IS
NONRESOLVING WITHOUT AN
INTERVENTION.
SO WE THINK THAT ACE INHIBITORS
AND ANGIO TENSEIN RECEPTOR
BLOCKERS CAN SUPPRESS THE
MICROTBLEEL ACTIVATION AND THEY
DO THIS IN PART BY INCREASING
LAYER 1 EXPRESSION.
SO C1 Q IS A PIVOTAL MOLECULE
HERE, IT CAN EITHER EXACERBATE,
SUSTAIN THE INJURY, BY BINDING
SCAVENGER RECEPTORS AND
MEDIATING PRUNING OF NEURONAL
DENDRITES OR IT CAN BIND TO
LAYER AND SUPPRESS THE
MICROGLIAL ACTIVATION AND ALLOW
THE NEURON TO RESTORE NORMAL
DENDRITIC ARBORIZATION EMPLOY WE
THINK THIS PARADIGM MAY BE
PRESENT IN MANY DISEASES, WE
THINK IN FACT IT IS THE PRESENT
IN ALZHEIMER'S DISEASE AND CAN
CONTRIBUTE TO CHANGES IN
COGNITION AND BEHAVIOR.
AND 1 OF THE THINGS THAT IS MOST
CURIOUS TO ME THAT THE PHENOTYPE
OF OF THE MICROGLIAL CELL THAT
IS RESPONSIBLE IN PART FOR THIS
BRAIN INJURY IS HIGH PRODUCTION
OF C1 Q, HIGH PRODUCTION OF
IL10, AND INCREASED CREASED CREC
POTENTIAL.
I THINK IT WAS ON 1 OF THE
SLIDES INCREASED EXPRESSION OF
AXLE WHICH IS 1 OF THE PH
AGOSITIC RECEPTORS ON
MACROIFIESS.
SO IF YOU THINK ABOUT THAT,
THAT'S THE PHENOTYPE OF THE PRO
RESOLVING MACROPHAGE IN THE
PERIPHERY.
AND IN FACT, AN M2 LIKE
MACROPHAGE WHICH WE THINK OF AS
ANTIINFLAMMATORY PRO
INFLAMMATION RESOLUTION HAS
INCREASED C1 Q PRODUCTION,
INCREASED IL10, AND INCREASED PH
AGOSITIERK C POTENTIAL.
SO I'M NOT SURE EXACTLY HOW TO
UNDERSTAND HOW 1 IS
ANTIINFLAMMATORY IN THE
PERIPHERY, AND PRO INFLAMMATORY
IN THE BRAIN, BUT I THINK IT
RAISES A VERY IMPORTANT CAVEAT
AS WE THINK ABOUT THERAPEUTICS
THAT FOCUS ON THE MYELOID CELL
BECAUSE WHAT YOU WOULD WANT TO
ENIENDER IN THE PERIPHERY MAY
ACTUALLY NOT BE GOOD FOR THE
BRAIN.
SO I'M GOING TO END THERE AND
JUST ACKNOWLEDGE THAT AS ALWAYS
IT TAKES A VILLAGE AND THERE
HAVE BEEN MANY WONDERFUL PEOPLE
BOTH FROM WITHIN MY OWN LAB,
MOST RECENTLY MARK AND KATE
WORKING ON THIS AND MANY
COLLABORATORS AT THE FINE STEIN,
DAVID, AND [INDISCERNIBLE] AND
CHRIS IN THE IMAGING STUDYING
AND BRUCE VOLPE AND TRUTH IN
ADVERTISING THESE MY HUSBAND IN
SOME OF THE HISTOLOGY OF THE
BRAIN.
CINDY AND MEGAN SEE THE PATIENTS
AND PAT O DOES THE PHYSIOLOGY
FOR US AND THEN WE HAD
COLLABORATORS AT OTHER
INSTITUTIONS WHO HAVE EITHER
HELPED US WITH EXPERIMENTS,
TAUGHT US SOMETHING OR GIVEN US
IDEAS.
I'M FLAD TO --GLAD TO ANSWER AY
QUESTIONS.
[ APPLAUSE ]
NWONDERFUL SEMINAR.
I WAS CURIOUS FOR THE CLINICAL
TRIAL WHY YOU CHOSE THE FDG PET
OVER THE ALLELE ACTIVATION?
>> YOU KNOW YOU HAVE TO CHOOSE 1
THING, AND WE HAD THE DATA THAT
PEOPLE WHO WERE ON CENTRALLY
ACTING ACE INHIBITORS OVER TIME
HAD A DECREASE IN THEIR HYPER
METABOLISM.
SO WE KNEW THAT.
I BELIEVE THEY WILL ALSO HAVE A
DECREASE IN MICROFLEEL
ACTIVATION, BUT WE DIDN'T HAVE
LONGITUDINAL STUDIES AT THE TIME
THAT WE WROTE FOR FUNDING FOR
THE TRIAL TO SHOW THAT THERE WAS
A DECREASE IN THOSE INDIVIDUALS
WITH THE MICROFLEEL ACTIVATION
BECAUSE WE JUST STARTED USING
THAT PET LIGAND LATER IN THE
COURSE OF OUR STUDIES.
SO I ACTUALLY THINK THAT YOU
WANT TO GO FOR THE TARGET RIGHT?
YOU WANT TO SHOW YOU HAD AN
EFFECT ON YOUR PRESUMED TARGET
AND HOPE THAT THE MICRODPLEEL
ACTIVATION WILL SURELY CHANGE
AND IF I WERE DESIGNING IT NOW
AND WE HAD LONGITUDINAL DATA
THAT WOULD BE THE PRIMARY END
POINT.
>> ALSO YOU MENTIONED THE C1 Q
LOSS OF FUNCTION IS A MAJOR
SUSCEPTIBILITY FOR SLE, IT SEEMS
LIKE THAT'S THE OPPOSITE OF WHAT
YOU JUST SHOWED US.
>> SO I'M ONLY SHOWING YOU IN
THE BRAIN, RIGHT AND BECAUSE IN
FACT, C1 Q IS CRITICAL TO
DECREASING MYELOID CELL
ACTIVATION IN THE PERIPHERY, AND
YOU GET ACTIVATED DENDRITIC
CELLS, ACTIVATED MYELOID CELLS,
MORE T-CELL ACTIVATION, MORE
B-CELL ACTIVATION AND YOU END UP
WITH A LUPUS LIKE PHENOTYPE, I
THOUGHT YOU WERE GOING TO ASK ME
WHAT THOSE PEOPLE LOOK LIKE IN
TERMS OF BRAIN FUNCTION AND I
WISH I KNEW.
KEITH ELCON AT THE UNIVERSITY OF
WASHINGTON IN SEATTLE HAS A
COHORT OF THEM AND I KEEP ASKING
HIM TO STUDY THE BRAIN.
>> THANK YOU .
>> SO I DON'T KNOW.
>> YES.
>> GREAT TALK BOO I THE WAY, IN
YOUR TALK YOU KEEP MENTIONING C1
Q BUT IN THE BRAIN IS C1 Q
COMPLEXED WITH C1 R AND C1 S AS
A COMPLEX OR IS IT STAND ALONE
NYES AND INFACT HGMGB1 INCREASES
PRODUCTION IN ALL OF THIS.
>> SO IN THE BRAIN IT WILL
ALWAYS--
>> OH, ALWAYS, --I DON'T KNOW.
>> I CAN TELL YOU THEY'RE THERE.
I CAN'T TELL YOU THAT THEY'RE
ALWAYS COMPLEXED TOGETHER.
AND I CAN TELL YOU C1 Q, ALONE
CAN BIND LAYER 1 AND THE WHOLE
COMPLEX CAN BIND LAYER 1 NSO THE
WHOLE COMPLEX IS NOT NECESSARY
FOR THIS PATTERN?
>> FOR ENGAGING LAIR 1 AND
SUPPRESSING MACROPHAGES IN THE
PERIPHERY, NO.
THE WHOLE COMPLEX IS NOT
NECESSARY.
>> DOES ANYTHING HAPPEN DOWN
STREAM OF THAT?
OR DOES IT END THERE?
>> YOU MEAN WHAT'S DOWN TREME OF
LAIR 1 IN LAIR 1 HAS 2 ITEM
MOTIFS.
>> NO, C1 Q?
>> WHAT?
>> IN THE NORMALY COMPLEMENT
ACTIVATION, DOES ANYTHING HAPPEN
AFTER THAT OR DOES THIS PATHWAY
JUST SORT OF END AT C1 Q AND
THEN ACTIVATING ON LAIR 1 NWELL,
C1 Q HAS MANY RECEPTORS THAT
REQUIRE ONLY C1 Q FOR ENGAGEMENT
AND DOWN STREAM PATHWAY
ACTIVATION.
C1 Q IS ALSO PART OF THE
COMPLEMENT CASCADE THAT CAN END
UP WITH THE MEMBRANE ATTACK
COMPLEX, RIGHT?
SO, I HONESTLY THINK OF HMGB1
AND C1 Q, AS THE PRIMORDIAL
CYTOKINES.
THEY'RE EVOLUTIONAR VERY
CONSERVED, THEY PRECEDE ADAPTIVE
IMMUNITY BY A GAZILLION YEARS
AND HMGB1 IS PRO INFLAMMATORY
AND AT LEAST IN THE PERIPHERY C1
Q IS ANTIINFLAMMATORY BUT C1 Q
CAN FUNCTION AS PART OF THE--AND
THEY BOTH HAVE NUMEROUS
RECEPTORS, SO IT'S BEFORE YOU
GOT 1 CYTOKINE, 1 RECEPTOR AND
IT CAN FUNCTION INDEPENDENTLY OF
THE COMPLEMENT CASCADE BINDING
MANY DIFFERENT RECEPTORS ON
CELLS.
>> AWESOME.
AND THEN MY SECOND QUESTION IS--
>> CAN YOU JUST STAY THERE
AND--OKAY, SORRY.
>> SO DID I GET IT RIGHT THAT
THE ANTIBODY INCREASES 10 FOLD
AFENNITY FOR DNA, AFTER IT GETS
THE MUTATION WHICH BINDS THE
RECEPTOR?
>> THE ANTIBODY WITH 10 FOLD
INCREASED AFFINITY FOR DNA IN
FACT DOES NOT CROSS REACT WITH
THE NMDA RECEPTOR.
IT'S THE LOWER AFFINITY AND THE
ANTIBODY WHICH WE GOT TO, WE GOT
TO THE PEPTIDE LIBRARY BECAUSE
WE WANTED TO KNOW THE DIFFERENCE
THEEN THOSE ANTIBODIES, BUT THE
ANTIBODY, THIS IS THE LOWER
AFFINITY ANTIBODY AND THIS IS
THE ANTIBODY THAT IS PRESENT IN
PATIENTS.
>> YEAH, SO DO YOU SEE IN THE
BRAIN, DO YOU SEE DNA RELEASE BY
DAMAGE CELLS WHICH THEN CAN
POTENTIALLY CUT OFF LINK AND
LEAD TO THIS SUSTAINING
INFLAMMATION AND CASCADE IN.
>> SO WE DON'T SEE INCREASE--
>> EXCUSE ME?
>> TALK TO THE MICROPHONE.
>> OH SORRY.
>> SORRY.
NWE DON'T SEE CONTINUAL LOSS OF
NEURONS IN THE BRAIN.
WE SEE LOSS OF NEURONS OVER THE
FIRST 48, 72 HOURS, AND THEN
WHEN WE GO COUNT NEURONS, WE
DON'T SEE ANY CONTINUOUS LOSS
AND WHEN WE LOOK AT THE BRAINS A
YEAR LATER, WE DON'T SEE THAT
THEY'RE MORE NEURONS LOST THAN
THERE WERE IMMEDIATELY.
SO WE CAN'T TELL THAT YOU
THERE'S NO DEATH IN RELEASE OF
DNA, BUT I DON'T THINK IT'S AN
ONGOING BIG EVENT.
YOU KNOW THERE'S SUPPOSED TO BE
NEURONAL NEOGENESIS AND WE
LOOKED FOR THAT AND WE DON'T SEE
THAT SO IT'S NOT AS IF NEW 1S,
IT'S NOT OBVIOUS TO US OR NOT
DETECTABLE BY US THAT NEW 1S ARE
BEING GENERATED AND OTHERS
CONTINUE TO DIE.
>> AND THE RECEPTOR IS IT
SPECIFIC FOR HYPOCAMPICAL
RECEPTOR OR NOT?
>> IS THE NMDA, IT'S ALL OVER
AND IF YOU INYECT, IT'S
WHERE--SO WE'VE DONE THE STUDY
WHERE WE GIVE MICE EPINEPHRINE
AND AT THE TIME OF THE FIRST
GULF WAR THEY DORPHED THAT
EPINEPHRINE OPENS THE BLOOD
BRAIN BARRIER AND BECAUSE OF ALL
OF THE TROOPS WHO GOT SOME YOU
KNOW DRUG TO PROTECT YOU AGAINST
GERM WARFARE AT BASE CAMP THEN
GOT PSYCHOTIC WHEN THEY GOT THE
DRUGS IN TANKS ON THE DESERTS OF
KUWAIT AND YOU GIVE EPINEPHRINE,
YOU OPEN UP THE BLOOD BRAIN
BARRIER IN THE ARK MIG DUALA
DUALA AND YOU GET AN ALTERATION
IN FEAR CONDITIONING.
>> SO WHY DO YOU THINK THEN IT
JUST TRIGGER THIS IS SPECIFIC
NEURONS IN THIS DEC, DO YOU HAVE
A HYPOTHESIS FOR THAT?
>> I THINK WHERE IT ENTERS THE
BRAIN IT CAUSES EXCITE O
TOXICITY OF THE NEURONS IN THAT
REGION.
IF YOU INYECT --INJECT IT INTO
THE CORTEXSTER O PATHICALLY YOU
GET DEATH IN THE CORTEX.
SO I THINK WE HAVE NOT AT ALL
SEEN EVERYWHERE IT MAY ENTER THE
BRAIN.
>> SO IN YOUR TALK YOU SAID THAT
LIKE PATIENTS ARE AT INCREASED
RISK OF NEUROPSYCHIATRIC
SYMPTOMS IF THEY HAVE SOME KIND
OF DISRUPTION TO THE BLOOD BRAIN
BARRIERS SO I WOULD AT LEAST
MEAN SOMETHING LIKE INFECTION OR
LIKE, YOU KNOW?
>> WELL, SO, IT CAN BE
INFECTION, THAT'S FOR SURE.
IT CAN BE VERY GREAT STRESS.
I DON'T KNOW THE DEGREE OF
STRESS BUT IT CAN BE THAT.
SMOKING, AGING, HYPERTENSION ALL
DISRUPT THE AT THE BLOOD BRAIN
BARRIER AND 1 OF THE THINGS THAT
I WORRY ABOUT A LOT IN ALL OF US
IS THAT THE GUT MICROBI OTA
MAINTAINED THE BLOOD BRAIN
BARRIER, AND WE SHOWED WITH
PATTERSON YEARS AGO THAT
GERM-FREE MICE DON'T HAVE A
BLOOD BRAIN BARRIER AND THAT IF
YOU IMIF MICE, ADULT MICE
ANTIBIOTICS THEY LOSE THEIR
BLOOD BRAIN BARRIER, THEY LOSE
THEIR BLOOD BRAIN BARRIER IN THE
HIPPOCAMPUS AND THE INTERRHINAL
CORTEX, SO, YOU KNOW IF YOU HAVE
MORE INFECTIONS YOU GET MORE
ANTIBIOTICS, AND SO I DON'T KNOW
WHAT'S CAUSING IT IN INDIVIDUAL
PATIENTS BUT THERE ARE LOTS OF
INSULTS THAT COULD COMPROMISE
THE BLOOD BRAIN BARRIER.
>> DOES IT MATTER THE TIMING OF
THESE INSULTS?
>> IF YOU HAVE THE ANTIBODIES
THEY'LL GET IN.
>> SO EVEN IF THEY'RE IN
REMISSION, IF THAT COULD BE
OBSERVED?
>> OKAY, THAIRNGS.
>> REALLY BEAUTIFUL TALK, THANK
YOU.
I HAVE 2 QUESTIONS, 1 IS--SO
YOUR ANTIBODY IS AGONISTIC, IT
SIGNALS THROUGH THE NMDR
RECEPTOR.
>> IT'S CALLED A POSITIVE
ALOESTERRIC MODULATOR THIS, IS
WORK WE DID WITH COLLEAGUES AT
STONY BROOK SO IT WILL NOT
ACTIVATE THE NMDA RECEPTOR BY
ITSELF.
IT POTENTIATES THE ACTIVATION IF
GLUTAMATE OR NMDA IS THERE.
>> SO DO YOU THINK THAT THAT THE
HMGB1 RELEASE BECAUSE YOU
MENTIONED IT HAS TO COME FROM
NEURONS RATHER THAN FROM GLUE
MARIOUSA, SO DO YOU THINK THAT
IT'S PART OF SOME KIND OF
COMPENSATORY MECHANISM TO DOWN
REGULATE SYNAPSE SIS ON NEURONS
THAT ARE EXPRESSED OR EXCITED BY
TOXICITY?
>> YOU KNOW, I THINK THERE'S
PROBABLY A ROLE OF HMGB1 IN
MAINTAINING NORMAL SYNAPTIC
FUNCTION IN NEURONS BECAUSE IT
BINDS NEURONS.
IT CAN POTENTIATE, IT ALSO
POTENTIATES THE NMDA RECEPTOR.
WE ACTUALLY SEE MORE DENDRITIC
ARBORIZATION IN THE KNOCKOUT
MICE, SO I THINK IT MAY BE
INVOLVED IN NORMAL DEVELOPMENT
BUT I AM NOT A NEUROSCIENTIST
AND I AM ALREADY FAR AFIELD FROM
THINGS I KNOW.
SO I THINK YOU'RE RIGHT.
I THINK IT IS PLAYING A ROLE AND
IT PLAYS A ROLE NORMALLY, BUT
EXACTLY WHAT THAT ROLE IS, I
DON'T KNOW.
AND THE LAST QUESTION IS, I
DON'T THINK I HAD FULLY
UNDERSTOOD WHAT DO YOU MEAN BY
REMAINING INFLAMMATION, RIGHT?
SO YOU SHOWED THAT THERE IS NO
ANTIBODY THERE, SO, I'M ASSUMING
THERE IS NO HNG B1, THE HNG B1
CONTINUES TO BE SECRETED TO A
YEAR'S TIME.
>> BUT IS IT SECRETED MORE IN
THIS MICE THAN IN THE CONTROL
MICE IN.
>> OH, ABSOLUTELY, I SHOWED YOU
THE PICTURE OF, SOPHISTICATED WE
CAN'T SEE SECRETED HMGB1, WE SEE
CYTOPLASMIC HMGB1 AND WHEN IT
LEAVES TO THE NUCLEUS TO GO TO
THE CYTOPLASM IT'S ACETALATED
AND RIGHT AND THEN SECRETED NYOU
SHOULD BE ABLE TO MEASURE HMGB1
IS SEE A SAMPLE OF THIS.
>> YEAH, YEAH, WE PROBABLY COULD
GETTING CSF OUT OF MICE WE'VE
DONE, IT'S NOT EASY AND
MEASURING HMGB1 IS ALSO NOT A
HIGHLY SENSITIVE ASSAY.
BUT YOU'RE RIGHT.
WE SHOULD DO IT.
>> AND YOU SEE THE CYTOPLASMIC
HMGB1 IN NEURONS, NOT NOT IN
MICROGLIA.
>> IN NEURONS.
IN NEURONS.
>> THANK YOU.
>> SO I WAS JUST WONDERING ABOUT
1 THING, AND ARE ALL THESE
ANTIBODIES COMING IN FROM THE
OUTSIDE OR AFTER THE BLOOD BRAIN
BARRIER IS OPENED CAN THEY COME
IN AND SETTLE INSIDE THEMSELVES
AND PRODUCE ANTIBODY IN C2 AND
IF YES, ANYBODY USING RETUX MAB?
NETWORK SO YOU KNOW RETUX MAB
HAS NOT WORKED IN THE CLINICAL
TRIALS IN LUPUS, NOBODY, I
ACTUALLY ASKED GENENTECH TO DO
BRAIN STUDIES IN THOSE TRIAL
ANDS THEY DIDN'T.
SO I DON'T KNOW ANYTHING ABOUT
THE BRAINS OF PEOPLE WITH THAT.
WE ACTUALLY ARE PARTNERING WITH
A CAR T-CELL GROUP TO LOOK AT
BRAINS NOW.
BUT WE DON'T SEE ANY B-CELLS IN
THE BRAIN AND IF YOU LOOK AT
PEOPLE LIKE THIS, IF YOU LOOK