I'M RICHARD DAVEY THE DEPUTY
CLINICAL DIRECTOR OF NIAID AND
I'M HERE TO INTRODUCE
DR. CHERTOW THE HEAD OF THE
EMERGING PATHOGEN SECTION IN THE
CLINICAL CENTERS CLINICAL CARE
DEPARTMENT AND TENURE-TRACK
INVESTIGATOR IN THE NIAID'S
LABORATORY OF REGULATION.
MANY OF YOU KNOW DAN BUT BY WAY
OF BRIEF BACKGROUND, DAN DID HIS
UNDERGRADUATE WORK AT BOSTON
COLLEGE FOLLOWED BY MEDICAL
SCHOOL AT NORTHWESTERN DURING
WHICH TIME HE ALSO OBTAINED AN
M.P.H. FROM U.C. BERKELEY AND
DID TRAINING AT U.C. SAN DIEGO
AND WAS AT THE INTELLIGENCE
SERVICE AND SPENT TIME WITH THE
FLORIDA STATE DEPARTMENT DEALING
WITH OUTBREAK AND CONTROL IN
THAT STATE.
HE CAME TO NIH IN 2007 FOR HIS
CRITICAL CARE MEDICINE
FELLOWSHIP FOLLOWING WHICH HE
DID INFECTIOUS DISEASE
FELLOWSHIP AT JOHNS HOPKINS.
ONCE HE HAD COMPLETED THOSE
TRAININGS, DAD -- DAN EMBARKED
IN AN EXCITING CAREER BASICALLY
FOCUSSING ON EMERGING PATHOGENS
OF MAJOR PUBLIC HEALTH
IMPORTANCE AND OVER THE TIME
HE'S BEEN HERE THAT'S INCLUDED
WORK ON SEASONAL INFLUENZA AND
RECENTLY ON OTHER MAJOR
PATHOGENS OF CONCERN YOU MAY
KNOW DAN MOSTLY FROM HIS WORK ON
TWO MORE RECENT PATHOGENS EBOLA
AND COVID-19.
IN TERMS OF HIS WORK WITH EBOLA
DAN SPENT TIME OVER IN MONROVIA,
LIBERIA UNDER THE AUSPICES OF
DOCTORS WITHOUT BORDERS.
UPON HIS RETURN HE WAS A SENIOR
A MEMBER OF THE CRITICAL CARE
MEDICINE DEPARTMENT WHO WORKED
WITH NIAID IN CARING FOR THE
FOUR INDIVIDUALS WE WERE
PRIVILEGED TO MEDICALLY EVACUATE
TO PROVIDE CARE FOR.
ONE PATIENT WAS PARTICULARLY ILL
AND IN -- DAD IS A
TRANSCRIPTIONAL ANALYSIS TO
EBOLA INFECTION OVER THAT TIME
PUBLISHED IN SCIENCE
TRANSLATIONAL MEDICINE.
DAN HAS BEEN INVOLVED IN CARING
FOR AND RESEARCHING PATIENTS
WITH COVID-19 INFECTION.
HE WAS AMONGST THE SENIOR TEAM
IN CCMD WHO WORKED WITH US AT
NIAID TO BRING THE FIRST
PATIENTS WITH SARS COV2 TO THE
CLINICAL CENTER AND PROVIDE CARE
FOR THEM AND WAS INSTRUMENTAL IN
THE FIRST RESEARCH PROTOCOLS
INCLUDING THE ACT-T PROTOCOLS
THAT ULTIMATELY RESULTED IN THE
LICENSURE OF REMDESIVIR FOR
TREATMENT OF COVID-19 INFECTION.
TODAY'S TOPIC'S GOING TO BE
FOCUSSED ON ONE OF HIS MOST
IMPORTANT BODY OF WORK IN THIS
AREA WHICH WAS INSTRUMENTAL IN
ORGANIZING A CONSORTIUM OF LOCAL
INVESTIGATORS TO CONDUCT A STUDY
OF INDIVIDUALS WHO SUCCUMBED TO
COVID-19 AND FAMILIES AGREED TO
HAVE THEIR BODIES SENT FOR
AUTOPSY AND NUMEROUS TISSUE
SAMPLES WERE OBTAINED LOOKING AT
THE PRESENCE OF VIRUS THROUGHOUT
THE BODY AND THAT'S THE TOPIC OF
DAN'S TALK ENTITLED SARS COV2
INFECTION AND PERSISTENCE
THROUGH THE HUMAN BODY AND
BRAIN.
LET ME TURN THINGS OVER TO DAN
FOR THIS I THINK WILL BE AN
EXCITING LECTURE.
>> I APPRECIATE THE INTEREST
GROUP'S INVITATION FOR THE
OPPORTUNITY TO GIVE THE LECTURE.
I'LL BE DISCUSSING A
RESEARCH WITH SARS COV2 AN
PERSISTENCE AMONG THE HUMAN BODY
AND BRAIN AMONG FATAL AUTOPSY
I HAVE NO DISCLOSURES.
WE HAD OBJECTIVES TO DESCRIBE
THE BURDEN OF SARS COV2
INFECTION WITHIN VERSUS OUTSIDE
THE RESPIRATORY TRACT TO DEFINE
SARS COV2 CELLULAR TROPISM AMONG
THE BODY AND BRAIN AND IF IT
PERSISTS IN TISSUES AND IF SO
OVER WHAT INTERVAL AND TO
DETERMINE IF SARS COV2 EVOLVES
IN DIFFERENT ANATOMIC
COMPARTMENTS OF THE BODY.
I'LL START BY DESCRIBING OUR
STUDY DESIGN.
WE COLLABORATED WITH THE
UNIVERSITY OF MEDICINE IN THE
CENTRAL CAPITAL AND EASTERN
SHORE REGIONS STUDY RECRUITMENT
WAS INITIATED BY DEDICATED
UPSTREAM CLINICAL PARTNERS OFTEN
WITH SURGING NUMBERS ALERTED US
WITH COVID-19 DEATHS WITH
POTENTIAL PARTICIPATION IN THE
STUDY.
SPECIFICALLY CRITICAL CARE
PROVIDERS ALERTED US FROM
PATIENTS WHO DIED AND NEXT OF
KIN WHO EXPRESSED INTEREST IN
STUDY PARTICIPATION.
THE NEXT OF KIN WERE CONSENTED
BY MYSELF AND THE REMAINS WERE
TRANSFERRED TO THE NIH CLINICAL
CENTER WHICH WAS COORDINATED
THROUGH OUR TRULY AMAZING CLINIC
CENTER ADMISSIONS DEPARTMENT AND
THE TISSUE PROCUREMENT WAS
PERFORMED BY THE NATIONAL
INSTITUTE PATHOLOGY TEAM TO KEEP
THE POSTMOREM INTERVALS OF OUR
STUDIES WITHIN THE FOLLOWING
MORN.
WE PERFORMED A TOTAL OF 44
AUTOPSIES BETWEEN MARCH 2020 AND
MARCH 2021.
EACH AUTOPSY EACH IS DENOTED AS
SHOWN HERE.
YOU CAN SEE OUR CASES WERE WELL
DISTRIBUTED OVER THE FIRST YEAR
OF THE PANDEMIC.
IT'S IMPORTANT TO EMPHASIZE UP
FRONT THAT NONE OF THESE
INDIVIDUALS WERE VACCINATE AND
ALL WERE INFECTED BEFORE
VARIANTS OF CONCERN WERE WIDELY
CIRCULATING IN THE UNITED
STATES.
AND WE PRESERVED MORE THAN
10,000 SPECIMENS FOR DOWNSTREAM
ANALYSES ACROSS 85 DISTINCT
ANATOMIC SITES AND BODY FLUIDS.
PERTAINING TO THE RESULTS I'LL
SHOW ON THE SUBSEQUENT SLIDES
FRESH TISSUE AND BODY FLUIDS
WERE PRESERVED FOR DOWN STREAM
ANALYSIS BY OUR DROPLET PCR
ASSAY FOR SARS COV2 AND FOR SUB
GENOMIC RNA THOUGHT TO BE A
MARKER OF RECENT REPLICATION AND
HIGH THROUGHPUT AMPLIFICATION
SEQUENCING TO COMPARE THE SARS
COV2 SPIKE GENE WITHIN AND
ACROSS ANATOMIC COMPARTMENTS.
ADDITIONALLY FLASH FLUIDS AND
TISSUES WERE FROZEN TO LOOK FOR
VIRUS AND HISTO LOGIC
EXAMINATION AND SARS COV2
TROPISM.
I'LL NOW PROVIDE THE CLINICAL
FEATURES OF OUR COHORT.
IT WAS DIVERSE WITH A MEDIAN AGE
OF 62.4 YEARS, 29.5% WERE FEMALE
AND 41% BLACK AND 15% HISPANIC
OR LATINO.
PRE EXISTING CONTINUES WERE
PREVALENT WITH 77% OF OUR
PATIENTS HAVE TWO OR MORE
COMORBID CONDITIONS,
CARDIOVASCULAR DISEASE, CHRONIC
RESPIRATORY DISEASE AND
DIABETES, HYPERTENSION AND
OBESITY WERE MOST PREVALENT.
THE MEDIAN INTERVAL FROM SYMPTOM
ONSET TO DEATH WAS 18.5 DAYS AND
THE MEDIAN POSTMORTEM INTERVAL
WAS 22.2 HOURS.
MOST OF THE COHORT WERE
ADMINISTERED VASO PRESSERS,
ANTIBODIES AND ANTICOAGULATION
PRE-MORTEM.
77% RECEIVED INVASIVE MECHANICAL
VENTILATION AND 41% RECEIVED
RENAL REPLACEMENT THERAPY PRIOR
TO DEATH.
THIS SECTION WILL FOCUS ON OUR
CHARACTERIZATION OF SARS COV2
TISSUE DISTRIBUTION AND
PERSISTENCE.
WE DEFINE PERSISTENCE AS
DETECTION OF ALL RNA IN TISSUES
GREATER THAN OR EQUAL TO 31 DAYS
AFTER INITIAL COVID-19 SYMPTOM
ONSET.
FOR EASE OF PRESENTATION, I'M
GOING TO SHOW A HEAT MAP
DEPICTING OUR SARS COV2 RNA
RESULTS LIMITED ONLY TO THE 11
PATIENTS FROM WHICH WE COLLECTED
BOTH BODY AND BRAIN TISSUES.
TO ORIENT YOU ON THE LEFT SIDE
ARE THE TISSUES SAMPLED
ORGANIZED BY TISSUE GROUPS.
THESE GROUPS INCLUDED
RESPIRATORY, CARDIOVASCULAR,
LYMPHOID AND GENERAL URINARY AND
SKELETAL MUSCLE AND TISSUES AND
EACH COLUMN REPRESENTS A PATIENT
WITH THEIR I.D. NUMBER AT THE
TOP AND DENOTED AT THE BOTTOM IS
THEIR DURATION OF ILLNESS PRIOR
TO DEATH.
ILLNESS DURATION IS GROUPED INTO
EARLY, MID AND LATE CASES
DEFINED BY DEATH WITHIN 14 DAYS
BETWEEN 15 TO 30 DAYS OR GREATER
OR EQUAL TO 31 DAYS FOLLOWING
SYMPTOM ONSET RESPECTIVELY.
DARK RED REPRESENTS HIGH NUMBERS
OF SARS COV2 NUCLEOTIDES AND
DARK BLUE REPRESENTS LOW COPY
NUMBERS BY RDPRR ASSAY.
AS YOU CAN SEE WE'VE TAKEN SARS
COV2 RNA ACROSS ALL TISSUE
GROUPS WITH THE HIGHEST LEVELS
DETECTS IN RESPIRATORY AND
CARDIOVASCULAR TISSUES AMONG THE
EARLY GROUP.
ADDITIONALLY, WE OBSERVED
RELATIVELY HIGH LEVELS OF SARS
COV2 RNA IN P33 OR PATIENT 33 IN
THE LATE GROUP AT 76 DAYS AFTER
ILLNESS ONSET AND NOTABLY
DETECTED SARS COV2 RNA IN
MULTIPLE TISSUES IN P42 SHOWN
ALL THE WAY TO THE RIGHT WHO
DIED APPROXIMATELY EIGHT MONTHS
AFTER COVID-19 ILLNESS ONSET.
IN THIS PARTICULAR CASE THE
PATIENT REPORTED ONLY MILD
COVID-19 ILLNESS RECOVERING AND
THEN CONTRACTED HEPATITIS A
ANDLET LED TO LIVER TRANSPLANT
AND ULTIMATELY DEATH.
DISPLAYED ARE THE PDCCR RESULTS
FOR THE SAME PATIENTS ACROSS 10
DISTINCT ANATOMIC SITES ACROSS
THE NERVOUS SYSTEM AND DETECTED
RNA IN ALL REGIONS TESTED AND
DETECTED RESIDUAL RNA IN ALL
LATE CASES BOXED IN RED
INCLUDING ACROSS MULTIPLE
REGIONS OF THE BRAIN AND P42 ALL
THE WAY TO THE RIGHT AT 230 DAYS
AFTER INITIAL INFECTION.
IN ORDER TO MORE FULLY
CHARACTERIZE CRENS AND RNA
BURDEN WITHIN VERSUS OUTSIDE THE
RESPIRATORY TRACT, WE FORMED A
LINEAR REGRESSION OF RNA LEVELS
IN RESPIRATORY VERSUS
NON-RESPIRATORY TISSUES BY
ILLNESS DAY.
ON THIS FIGURE LOG TRANSFORMED
RNA LEVELS ARE SHOWN ON THE Y
AXIS AND DAY OF ILLNESS ON THE X
AND RNA LEVELS WERE
SIGNIFICANTLY HIGHER IN THE
RESPIRATORY TISSUES INDICATED BY
THE RED COLOR COMPARED TO
NON-RESPIRATORY TISSUES SHOWN IN
BLUE FOR MONTHS FOLLOWING
ILLNESS ONSET.
A SIGNIFICANTLY STEEPER RATE OF
DECLINE IN RNA LEVELS OR
NEGATIVE SLOPE WAS OBSERVED IN
RESPIRATORY COMPARED TO
NON-RESPIRATORY TISSUES.
THE Y SLOPE VALUES ARE BOTH
GROUPED AND INDIVIDUAL
RESPIRATORY AND NON-RESPIRATORY
TISSUES ARE SUMMARIZED IN THE
TABLE TO THE RIGHT.
WE ALSO PERFORMED A LINEAR MIXED
MODEL ANALYSIS OF RNA BURDEN IN
RESPIRATORY VERSUS
NON-RESPIRATORY TISSUES IN OUR
PRE-DETERMINED EARLY, MID AND
LATE GROUPS.
AMONG THE EARLY GROUP WE
OBSERVED APPROXIMATELY 100 FOLD
HIGHER SARS COV2 LEVELS IN
RESPIRATORY COMPARED TO
NON-RESPIRATORY TISSUES.
A 20 FOLD AND FOURFOLD
DIFFERENCE IN THE MID AND LATE
GROUPS RESPECTIVELY.
HERE'S TO THE SAME HEAT MAP WE
LOOKED AT BEFORE BUT NOW WITH
THE RESULTS FROM THE QPCR
EVALUATING FOR SUBGENOMIC RNA
DENOTED BY VERTICAL HASH LINES
OVERLYING OUR COLOR-CODED HEAT
MAP.
I'D LIKE TO FIRST HIGHLIGHT THE
FOUR EARLIEST PATIENTS
REPRESENTING A DURATION OF UP TO
18 DAYS POST-ILLNESS ONSET ALL
HAD SUB GENOMIC DETECTED ACROSS
RESPIRATORY AND MANY
NON-RESPIRATORY TISSUES
INCLUDING THE BRAIN.
WE ALSO DETECTED SUBGENOMIC RNA
SPORADICALLY IN SOME LATE CASES
INCLUDING ACROSS FOUR TISSUE
GROUPS IN P33 AT 76 DAYS
FOLLOWING ILLNESS ONSET.
TO BETTER CHARACTERIZE THE
RELATIONSHIP BETWEEN SARS COV2
RNA LEVELS DETECTED BY OUR PDCCR
ASSAY AND LEVELS OF SARS COV2
SUB GENOMIC RNA DETECTED WE
CONDUCTED CORRELATIONS.
AMONG THE 10,025 SAMPLES THAT
WERE EVALUATED AND FOR SUB
GENOMIC RNA THE RNA LEVELS
SIGNIFICANTLY CORRELATED WITH
THE CORRELATED COEFFICIENT OF
76.
THIS ASSOCIATION STRENGTHENED TO
.88 AND . 86 WHEN THE ANALYSIS
WERE RESTRICTED TO THE 496
SAMPLES FROM EARLY CASES OR THE
396 SAMPLES COLLECTED FROM
RESPIRATORY TISSUE RESPECTIVELY.
WHEN THE ANALYSIS WAS LIMITED TO
THE 302 SAMPLES THAT WERE
POSITIVE FOR BOTH OF OUR ASSAYS,
A SIGNIFICANT CORRELATION
COEFFICIENT OF .91 WAS OBSERVED.
FINALLY, WE FORMED A REGRESSION
ANALYSIS USING THE QUANTITY OF
OF DETECTED BY PCR TO PREDICT
WHETHER A SAMPLE WOULD BE
POSITIVE AND THE PDCCR VALUE OF
1.47 COPIES PER NANOGRAM INTO
THE ASSAY RESULTED IN 93%
SENSITIVITY AND 91.6%
SPECIFICITY WITH AN RUC UNDER
THE CURVE OF .965 SUPPORTING THE
STRONG PREDICTIVE CAPACITY OF
THIS MODEL.
AFTER DETECTING SUBGENOMIC RNA
IN OVER 300 SAMPLES WE SOUGHT TO
DETERMINE IF WE COULD ISOLATE
SARS COV2 IN CULTURE AND WE
ATTEMPTED TO ISOLATE IT FROM
RESPIRATORY AND NON-RESPIRATORY
TISSUES ACROSS 20 PATIENTS.
WE SUCCESSFULLY COLLECT FROM
NINE PATIENTS WHO DIED FROM ONE
TO 12 DAYS OF ILLNESS ONSET AND
IN THE TISSUES CYTOPATHIC EFFECT
AND RELATIVE TO LEVELS IN THE
ORIGINAL TISSUE OBSERVED
CONFIRMING REPLICATION CONFIDENT
VIRUS.
WE FURTHER CHARACTERIZED THE RNA
AND REPLICATION CONFIDENT VIRUS
AND OBSERVED THAT 100% OF
TISSUES WITH A SUB GENOME
THRESHOLD BETWEEN 15 AND 20 HAD
REPLICATION CONFIDENT VIRUS.
THIS NUMBER DECREASED TO 64.3%
OF TISSUES THAT HAD A CT VALUE
BETWEEN 20 AND 25 AND FURTHER
DECREASED TO 31.1% OF TISSUES
THAT HAD A CT VALUE BETWEEN 25
AND 30 AND NO REPLICATION
COMPETENT VIRUS WAS DETECTED IN
TISSUES ACROSS THE THRESHOLD OF
30 OR HIGHER.
OVERALL, WE'RE ABLE TO ISOLATE
SARS COV2 FROM NASAL SINUS, LUNG
AND IN THE NON-RESPIRATORY
COMPARTMENT OF THE EYE, THE
MEDIAL STINAL LYMPH NODE AND
ADRENAL GLAND.
WE ESTIMATE SARS COV2 RNA LEVELS
STRONGLY PREDICTIVE OF
REPLICATION CONFIDENT OF
CULTURE.
TO ACCOMPLISH THIS WE FORMED
REGRESSION ANALYSIS WITH THE ROC
CURVES AND FOUND THAT A DVPCR
VALUE OF THE COPIES PER NANOGRAM
PUT IN THE ASSAY PREDICTED
REPLICATION COMPETENT VIRUS WITH
A 76% SENSITIVITY, 90%
SPECIFICITY AND ROCAUC OF .887.
A SUB GENOMIC RNA VALUE OF
25,069 COPIES PER MICROLITER OF
RNA PREDICTED REPLICATION
COMPETENT VIRUS WITH 72%
SENSITIVITY AND 100% SPECIFICITY
WITH AN AUCROC OF .915 AND
DETERMINE IT'S LIKELY
REPLICATION COMPETENT VIRUS
WITHOUT HAVING TO PERFORM CELL
CULTURE IN A BSL3 SETTING WHICH
IS A LIMITED RESOURCE.
NOW I'LL BRIEFLY TAKE YOU
THROUGH THE INSITU RESULT IN OUR
COHORT.
USING OUR DVPCR AND SUBGENOMIC
QPCR RESULTS AS A GUIDE, WE
REALIZED AN INSITU WITH THE
GENOME TO VALIDATE OUR PCR
ASSAYS AND POSITIVE NEGATIVE
CONTROLS FOR HUMAN HOUSEKEEPING
GENE AND E. COLI GENE WERE USED
TO IDENTIFY SPECIFICITY OF THE
ASSAY.
WHAT WE FOUND POSITIVE STAINING
IN ALL TISSUE GROUPS I HAVE A
SMALL PANEL THAT HIGHLIGHTS
STAINING IN BRAIN AND OTHER
TISSUES.
DISPLAYED IN THE FIRST ROW IN
ORDER FROM LEFT TO RIGHT ARE
FRONTAL LOBE OF THE LEFT AND
CORPUS CALLOSUM AND CEREBELLUM
OF A LATE CASE AND CERVICAL
SPINAL CORD OF A LATE CASE D.
SHOWN IN THE SECOND ROW ARE
FOLLICULAR CELLS AND THE SPLEEN
AND PANEL G AND EPITHELIUM AND
MONONUCLEAR AND EXCUSE ME SPLEEN
IS G AND IN H COLONIC
EPITHELIUM AND HERE'S THE
ADRENAL GLAND A AND THE TESTES
IS K AND THE STROMAL CELLS OF A
UTERUS IN L.
WE FURTHER EVALUATED SARS COV2
CELLULAR TROPISM WITHIN THE
CEREBELLUM AND OTHER REGIONS OF
THE BRAIN WITH FOCAL MICROSCOPY.
AND THE IMAGES THE CELL NUKE LEE
ARE BLUE AND THE NUCLEO CAPSID
IS GREEN AND NEURONS ARE
MAGENTA.
THE LOCATIONS OF THE MOLECULAR
LAYER AND GRANULAR LAYER AND
WHITE MATTER ARE INDICATED ON
THE SLIDE.
PANEL A SHOWED A LOW
MAGNIFICATION VISUALIZATION OF A
KNOWN ANTIBODY CONTROL AND B
SHOWS AN ADJACENT SECTION WITH
SPECIFIC PROTEIN EXPRESSION.
HIGHER MAG ANY FINGS --
MAGNIFICATIONS DEMONSTRATES
SPECIFIC INFECTION IN C AND
POSITIVE NEURONS ARE INDICATED
IN THE YELLOW ARROWS.
OTHER UNIDENTIFIED CELLS ARE
ASSOCIATED WITH PROTEIN IN THE
GRANULAR LAYER.
KIDNEY CELLS ARE INEFFEFECTED AN
E THEY WERE ASSOCIATED WITH
VIRAL PROTEIN.
THIS SUMMARIZES THE CELL TYPES
ACROSS THE BODY AND BRAIN THAT
WERE ISH POSITIVE FOR SARS COV2
IN OUR COHORT.
WE LOOKED AT DIFFERENT ANATOMIC
COMPONENTS WITHIN THE PATIENT
AND THE BOARD'S LAB PERFORMED
HIGH THROUGHPUT SINGLE GENOME
SEQUENCING FROM A TOTAL OF 46
TISSUES FROM SIX INDIVIDUALS.
AND PATIENT 18 ARE SHOWN ON THE
LEFT WHICH WERE IDENTICAL ACROSS
ALL ANATOMIC COMPARTMENTS
TESTED.
P19 ON THE RIGHT IN THIS PATIENT
WE DETECT A SINGLE SYNONYMOUS
MUTATION WHICH IS DENOTED IN
BLUE HERE AND A MINOR VARIANT IN
THE WHITE SUPERIOR LUNG LOBE.
OVERALL THE TWO PATIENTS
DEMONSTRATE SARS COV2 CAN INFECT
CELLS WITHIN NON-RESPIRATORY
TISSUES WITHOUT REQUIRING
CHANGES IN THE SPIKE GENOME.
AND SYNONYMOUS MUTATIONS WERE
DETECTED WITHIN THE RESPIRATORY
TRACT AND OUTSIDE OF IT AND THE
MEDIAL STINAL LYMPH MODE AND
DENOTED IN BLUE.
WE WERE UNABLE TO GET TISSUE
FROM THE LATE CASE THERE ARE TWO
TWO TYPES DETECTED WITHIN THE
THORACIC LYMPH NODE OF THIS
INDIVIDUAL.
FINALLY, WE'RE ABLE TO SEQUENCE
BRAIN SAMPLES FROM TWO PATIENTS
IN OUR COHORT WHICH ARE SHOWN
HERE WHICH REVEALED
NON-SYNONYMOUS MUTATIONS WITHIN
HAPLOTYPES WITHIN THE CNS AND
THE REST OF THE BODY
SPECIFICALLY IN THE DURAMATTER
FOR P36 AND THE HYPOTHALAMUS AND
THALAMUS OF P38 SHOWN HERE.
THESE RESULTS SHOW SARS COV2
WITHIN THE BRAIN IS SUBJECT TO
THE DEVELOPMENT OF GENETIC
COMPARTMENTALIZATION AND
EXPLORING THE MECHANISMS BEHIND
THIS WILL BE IMPORTANT FOR
FUTURE WORK.
I'LL SUMMARIZE THE HISTO LOGIC
CORRELATES IN OUR COHORT.
OUR PATHOLOGY COLLEAGUES AT NCI
IN ADDITION TO RUNNING ALL OF
OUR ISH ASSAYS DETERMINED THE
CAUSE OF DEATH AND CHARACTERIZES
THE PATHOLOGIC FINDINGS FOR ALL
CASES.
THE DAMAGE WAS DETECTED IN ALL
BUT 18% OF CASES AND A PANEL
SHOWS THE DAMAGE WITH MEMBRANES
AND PANEL B SHOWS THE
PROLIFERATIVE PHASE WITH SPARSE
INFLAMMATION IN A MILD CASE.
EXCUSE ME, NOT MILD BUT MID
CASE.
ACUTE PNEUMONIA WAS PRESENT IN
61% AND PULMONARY HEMORRHAGE IN
32% DEMONSTRATED DIFFUSELY IN
PANEL B AND THROMBOWLISM WAS
DETECTED IN 23% AN EXAMPLE IN
PANEL C IN A MEDIAN PULMONARY
VESSEL ON THE RIGHT.
WE WERE ABLE TO QUANTIFY THE
PROGRESSION FROM THE EARLY PHASE
SHOWN BY THE ORANGE STACK BARS
DURING WEEK 1 TO 2 TO THE
INTERMEDIATE PROLIFERATIVE PHASE
THE GRAY STACK BARS IN THE LATE
FIBROSING PHASE.
IN THE HEART WE SAW INFILTRATED
IN 9% DESPITE RNA IN 80% OF OUR
CASES.
PATIENT 3 SHOWN IN PANEL E IS AN
EXAMPLE OF A SINGLE CASE THAT
HAD ACTIVE LYMPHOCYTIC CARDITIS
AT THE TIME OF DEATH AND
ISCHEMIA AND MYOCARDIAL
MYCROISIS WAS DENOTE AND LYMPH
NODE ARCHITECTURE REMAPED INTACT
WITHOUT LYMPHODEFLATION --
LYMPHODEPLETION AND THERE WAS
WELL DEFINED FOLLICLES IN
CYTOSIS.
IN CONTRAST, SOME DEGREE OF
LYMPHO DEPLETION WAS SCREENED IN
35% OF CASES PANEL K IS LYMPHO
DEPLETION HERE.
WE OBSERVED NON-SPECIFIC
FINDINGS INCLUDING FOCAL OR
DIFFUSE HYPOXIC ISCHEMIC INJURY
IN 45% OF CASES.
VASCULAR CONGESTION WAS OBSERVED
IN 45% OF CASES AND MICROSCOPIC
HEMORRHAGE IN 18% OF CASES.
IN SUMMARY, WE OBSERVED THAT
SARS COV2 IS WIDELY DISSEMINATED
ACROSS THE HUMAN BODY AND BRAIN
EARLY IN INFECTION AND SARS COV2
RNA CAN PERSIST IN MULTIPLE
ANATOMIC COMPARTMENTS FOR OVER
SEVEN MONTHS AND OBSERVED THE
APAUCITY -- APOSITY OF
INFLAMMATION AND I'LL MAKE MANY
IMPORTANT ACKNOWLEDGEMENTS.
I'D LIKE TO START WITH THE MANY
HEALTH CARE PROVIDERS AT OUR
CONSORTIUM HOSPITALS LISTED
HERE.
I'D LIKE TO SPECIFICALLY
ACKNOWLEDGE DR. ALISON GRAZIOLI
WHO SPEARHEADED THE PATIENT
ENROLLMENT AT ALL OF OUR
CONSORTIUM HOSPITALS AND TO
ACKNOWLEDGES DR. SAHARIA AND
DR. RABIN INVOLVED IN THIS
COHORT.
THIS WORK WOULD NOT HAVE BEEN
POSSIBLE WITHOUT THE
ENTHUSIASTIC LEADERSHIP AND
SUPPORT BY DR. HEWITT AND MANY
IN THE LABORATORY MEMBER
SUPPORTING THE CULTURE WORK I
SHOWED AND DOCTORS PETERSON AND
CLAYTON WINKLER WHO LED THE WORK
IN THE BRAIN.
DR. BORITZ GROUP PERFORM THE
SPIKE SEQUENCING AND I RECOGNIZE
DR. HOLLAND FOR ENCOURAGEMENT
AND FUNDING TO SUPPORT THE
STUDIES EARLY ON AND CONTINUALLY
THROUGH THIS PANDEMIC.
MANY MEMBERS OF THE MEDICINE
DEPARTMENT CONTRIBUTED TO THIS
WORK DIRECTLY OR INDIRECTLY.
I'D LIKE TO SPECIFICALLY
ACKNOWLEDGE DR. STEIN THE FIRST
AUTHOR OF THIS WORK AND SABRINA
ROMELI WHO WORKED THE PDCCR ON
THOUSANDS OF SPECIMENS AND LIKED
TO THANK OUR STATISTICS SUPPORT
WHO PERFORMED OUR REGRESSION
ANALYSES.
I'D LIKE TO ACKNOWLEDGE AND
THANK TRACY LEWIS AND ALL
MEMBERS OF THE NIH CLINICAL
CENTER MISSION DEPARTMENT STAFF
FOR THEIR AGILITY AND
ADAPTABILITY IN SUPPORTING OUR
REQUESTS FOR TRANSFERS OF NEXT
OF KIN OR DECEASED OFTEN IN THE
EARLY HOURS OF THE MORNING
DURING THIS ONE-YEAR INTERVAL
OVER AND OVER AGAIN.
AND WITH THAT, I WILL THANK YOU
AND CLOSE AND TAKE QUESTIONS.
>> THANK YOU VERY MUCH FOR A
VERY INTERESTING TALK.
WE HAVE A LOT OF QUESTIONS BUT
BEFORE I START WITH THEM I WANT
TO REITERATE THE CME CODE NUMBER
FOR THOSE TAKING THE CREDIT.
IT'S 37971.
37971.
I'LL START WITH A QUESTION FROM
JIM BORIS.
DO POSTMORTEM ANALYSIS OF PEOPLE
WHO HAVE HAD MILD COVID-19 AND
RECOVERED FROM IT AND DIED OF
SOMETHING ELSE AFTER DO THEY
SHOW THE SAME CELLULAR TROPISM
AND PERSISTENCE OF SARS COV2?
>> EXCELLENT QUESTION, JIM.
WE'RE VERY CAUTIOUS AND OPENLY
ACKNOWLEDGE THAT OUR COHORT WAS
LARGELY LIMITED TO OLDER
INDIVIDUALS WITH A NUMBER OF
COMORBID CONDITIONS THAT
DEVELOPED SEVERE AND ULTIMATELY
FATAL COVID SO WE'RE CAUTIOUS
ABOUT GENERALIZING OUR
OBSERVATIONS TO A YOUNGER,
HEALTHIER POPULATION OR
VACCINATED.
THOSE ARE ALL LIMITATIONS.
HAVING SAID THAT, THERE ARE A
NUMBER OF INDIVIDUALS IN OUR
COHORT THAT WERE YOUNG.
WE HAD A 6-YEAR-OLD WHO
PRESENTED TO THE HOSPITAL FOR A
COMPLICATION OF A SEIZURE
DISORDER UNRELATED TO COVID AND
INCIDENTALLY FOUND TO HAVE COVID
WHERE WE FOUND THE SAME
DISTRIBUTION OF VIRUS ACROSS
THAT INDIVIDUAL'S BODY AND OF
COURSE I MENTIONED THE ONE CASE
THAT WAS IN AN ADULT WHERE THIS
PERSON HAD REPORTED MILD
RESPIRATORY SYMPTOMS AND EIGHT
MONTHS LATER UNRELATED TO COVID,
AS FAR AS WE KNOW, DEVELOPED
HEPATITIS A IN FUNCTION THROUGH
FOODBORNE ILLNESS AND DIED OF A
COMPLICATION 230 DAYS AFTER
THEIR INITIAL ILLNESS WE
OBVIOUSLY FOUND PERSISTENCE OF
RNA IN MANY COMPARTMENTS.
THE SHORT ANSWER IS WE'RE
CAREFUL ABOUT GENERALIZING OUR
OBSERVATION TO A DIFFERENT
POPULATION AND DO THINK THE
STUDIES NEED TO BE DONE.
>> HE'S ASKING WHETHER THE
HISTO PATH ARE MINIMAL OR LOWER
IN THE LATE CASES WHAT'S THE
CAUSE OF DEATH IN THE LATE
CASES.
>> IN THE SHORT ANSWER IS WE
HAVE A TABLE THAT SUMMARIZES
INDIVIDUAL-LEVEL PATIENT DATA
THAT WILL SPECIFICALLY OUTLINE
THE CAUSE OF DEATH FOR EACH
PATIENT BUT IN GENERAL THOSE ARE
OLDER AND HAD MORE COMORBIDITIES
MORE OFTEN THAN NOT WERE
ADMITTED TO THE HOSPITAL, REMAIN
IN THE HOSPITAL FOR AN EXTENDED
PERIOD OF TIME WITH PROVIDED
LIFE SUPPORT AND DIED DUE TO
COVID COMPLICATIONS TYPICALLY
RESPIRATORY FAILURE.
IN THAT GROUP WE OBVIOUSLY HAVE
A NICE TIME LINE OF WHEN
SYMPTOMS STARTED AND WERE
OBSERVED IN THE HOSPITAL FOR AN
EXTENDED PERIOD OF TIME AND HAVE
AN INTERVAL AT THE TIME OF
DEATH.
AS A MENTIONED, AN INTERESTING
FINDING WE'RE FOCUSSED ON AND
NEEDS TO BE FOCUSSED ON IS WHY
IS IT THAT THE VIRUS GETS TO
REMAIN IN PLAIN SIGHT IN ALL
THESE EXTRA PULMONARY SITES WITH
INJURY AND/OR INFLAMMATION.
HOW IS IT THE VIRUS LEADS THE
IMMUNE RESPONSE IN THE EXTRA
RESPIRATORY SITES.
>> QUICK QUESTION FROM ME BEFORE
I GET TO THE NEXT ONE.
THE PATIENTS WHERE YOU SAW BRAIN
INFILTRATION, WHERE FOR INSTANCE
THE CELLS WERE STAINING IT
WASN'T CLEAR TO ME, WAS A
STRUCTURAL STAINING OF THE
PROTEIN OF THE VIRUS OR
INDICATING LOW LEVEL REPLICATION
OF THE CELLS?
>> SO THE TARGET FOR OUR
IMMUNOHISTIC CHEMISTRY WAS NUKE
LEO CAP -- NUKE L-- NUCLEOCAPSI
AND WE APPROACHED THE BRAIN
BECAUSE THERE'S MAJOR
IMPLICATIONS ABOUT FINDING VIRUS
IN MULTIPLE REGIONS OF BRAIN AND
SPECIFIC CELL TYPES.
SO WE'RE QUITE CAUTIOUS ABOUT
THOSE OBSERVES AND HAVE TAKEN A
NUMBER OF ASSAYS TO SUPPORT OR
CONFIRM CELL TYPE SPECIFICITY.
AS FAR AS REPLICATION IN THE
BRAIN WE HAVE RIGHT NOW INDIRECT
AND PURSUING DIRECT EVIDENCE OF
REPLICATION COMPETENT VIRUS IN
THE BRAIN.
THE INDIRECT EVIDENCE IS THAT
THE SEQUENCING RESULTS I SHOWED
YOU, SHOW EVIDENCE IN THE BRAIN.
THEY'RE NEARING EQUAL
DISTRIBUTION OF VARIANCE WITH
NON-SYNONYMOUS MUTATIONS WE
DETECTED IN THE BRAIN THAT WE
DID NOT DETECT IN THE
RESPIRATORY TISSUE SUGGESTING
THAT ONGOING VIRAL REPLICATION
DOES TAKE PLACE IN THE BRAIN AND
THEY'RE ACTUALLY IS SOME
EVOLUTION OF THE VIRUS IN THE
BRAIN.
WE HAVE TRIED TO GROW THE VIRUS
FROM THE BRAIN BUT WE'RE NOT
DONE.
WE'RE GOING TO PERSIST.
SO FAR WE HAVE NOT DEFINITIVELY
ACCOMPLISHED THAT BUT SOMETHING
WE CONTINUE TO PURSUE.
>> AND YOUR ANSWER IS RELATED TO
TWO QUESTIONS SO LET ME ASK.
ONE IS FROM ROLAND OWENS, DOES
YOUR VIRUS DISTRIBUTION STUDY
SUGGEST ANY NOVEL RECEPTORS FOR
INSTANCE IN THE BRAIN WHAT'S THE
DISTRIBUTION OF THE CLASSICAL
RECEPTORS SERVICE WHAT MIGHT BE
NOVEL AND SECOND QUESTION IS
FROM CAROLYN OBARU, THANK YOU
FOR THE SEMINAR, CAN YOU PLEASE
BRIEFLY ELABORATE ON THE GENETIC
COMPARTMENTALIZATION YOU SEE IN
THE INFECTED BRAIN TISSUES.
WHAT DOES THAT MEAN?
YOU ALLUDED TO THAT SO MAYBE YOU
CAN REITERATE.
>> THAT'S PERFECT.
DR. OWENS, THANK YOU FOR YOUR
QUESTION AND ALSO FOR THE SECOND
QUESTION AS WELL.
SPECIFICALLY, WE DID NOT OR HAVE
NOT YET QUERY THE ASSOCIATION
WITH THE KNOWN SARS COV2
RECEPTORS AND CO-RECEPTORS IN
TISSUES AND CO-LOCALIZATION OR
PRESENCE OF VIRUS.
THAT WAS BY OUR ASSAYS.
LARGELY BECAUSE OTHERS HAVE
FOCUSSED ON THE PROTEOME MAP OF
THE HUMAN BODY OR PRESENCE OR
ABSENCE OF THESE RECEPTORS IN
DIFFERENT TISSUES AND SO WE
DIDN'T PURSUE THAT.
OBVIOUSLY, THAT IS SOMETHING
THAT WE COULD PURSUE WE CANNOT
DIRECTLY ANSWER THAT QUESTION
BUT THERE ARE CELL TYPES THAT
ARE NOT KNOWN TO HAVE ACE 2
RECEPTORS, MONOSIGHTSS --
MONOCYTES HAVE PRESENT AND IT'S
BEEN PUBLISHED IN NATURE THERE'S
EVIDENCE THOSE CELLS ARE EITHER
INFECTED OR PERHAPS TAKE UP
OPSENIZED VIRUS FOR THE ORIGINAL
SARS COV2 RECEPTORS AND THAT'S
WORK WE CAN PURSUE AND
ADDITIONAL WORK THAT LIKELY
NEEDS TO BE ACCOMPLISHED.
AS FAR AS THE
COMPARTMENTALIZATION OF THE
VIRUS, I SHOWED SIX CASES IN
THOSE TWO CASES, THE IMPORTANT
OBSERVATION AND THIS IS
SOMETHING THAT NEEDS TO BE
FOLLOWED UP WITH LARGER COHORTS
AND COHORTS THAT EMULATE OUR
STUDY DESIGN THAT OPTIMIZE
TISSUE PRESERVATION AND SHORT
POST MORTEM OBSERVATIONS WE HAD
BUT THE OBSERVATION IS THERE
WERE VIRAL VARIANTS IN THE
BRAIN.
POPULATIONS OF VIRAL VARIANTS IN
THE BRAIN NOT DETECTED IN THE
RESPIRATORY TRACT SUGGESTING
THAT THERE'S SEEDING OF VIRUS IN
THE BRAIN, THERE'S ONGOING
REPLICATION IN THE BRAIN AND AS
IT REPLICATES THERE'S A VARYING
POPULATIONS PRODUCED AND THROUGH
THE VERY SENSITIVE SEQUENCING
TECHNIQUE OUT OF THE LAB WE'RE
ABLE TO DETECT VARIANTS IN THE
BRAIN SUGGESTING THAT THERE IS
REPLICATION THERE AND PERHAPS
THERE'S A PARTICULAR IMMUNE
ENVIRONMENT THERE AND THIS IS
SPECULATIVE.
OBVIOUSLY, IT'S AN IMMUNE
PROTECT SITE THAT WILL ALLOW FOR
VIRAL REPLICATION AND EVOLUTION
OF THE VIRUS.
>> RELATED TO THAT AGAIN THERE'S
A LOT OF QUESTIONS ABOUT THE
BRAIN ACTUALLY.
WERE YOU ABLE TO QUANTIFY IN THE
CSF?
>> THE SHORT ANSWER IS I DON'T
KNOW I NEED TO PHONE A FRIEND
FROM DR. STEIN.
I KNOW AS TO WHETHER OR NOT WE
COLLECTED CSF.
IF WE COLLECTED CSF IT WAS ON A
LIMITED NUMBER OF INDIVIDUALS
AND WHETHER OR NOT WE DETECTED
RNA OR SUB GENOMIC RNA I CANNOT
RECALL.
I'D HAVE TO PHONE A FRIEND AND
CHECK THE PRE PRINT.
I DO KNOW WE DIDN'T PURSUE
VACCINATION OF ANTIBODY
RESPONSES OR FOR EXAMPLE SOLUBLE
MEDIATOR RESPONSES OR OTHERWISE
IN CSF.
I'D HAVE TO GO BACK TO REVIEW
THE INVENTORY TO DETERMINE WHAT
EXISTS THAT MIGHT BE
ACCOMPLISHED WITH THAT SAMPLE
TYPE.
>> HAVE YOU SEEN THE PRESENCE OF
VIRAL PARTICLES IN EYE TISSUES?
THANK YOU FOR THE QUESTION.
WE DID QUERY EYE TISSUE AND
FOUND PRESENCE OF RNA IN EYE
TISSUE SUMMARIZED AND
CATEGORIZED IN THE PRE PRINT.
WHAT'S NOT IN THE PRE PRINT ARE
CONFIRM TRY IMMUNO ISH CELL
TYPES IN THE EYES AND THAT MAY
OR MAY NOT BE FORTHCOMING IN A
SEPARATE BODY OF WORK.
>> THIS IS A QUESTION IS THERE
ANY INDICATION -- THIS IS VERY
SPECULATIVE RESIDUAL COV2 IN THE
BRAIN OR OTHER TISSUES CAN
BECOME RE-ACTIVATED TO PRODUCE A
DISEASE CONDITION IN THE FUTURE.
IT'S EARLY TO SAY, RIGHT?
WE'RE AT THE BEGINNING OF THIS.
>> YOU'RE CORRECT, YOU'RE ANSWER
IS THE CORRECT ANSWER.
WE CAN'T ANSWER THAT BASED ON
THE STUDY WE DID.
IT WASN'T DESIGNED AS SUCH BUT
FOR CERTAIN THOSE ARE THE TYPES
OF QUESTIONS THAT NEED TO BE
PURSUED IN THE FUTURE AND
OBVIOUSLY THERE ARE CONCERNS OR
QUESTIONS ABOUT, WELL, WHAT IS
THE RELATIONSHIP BETWEEN POST
ACUTE SEQUELAE AND OUR
OBSERVATION OF mRNA IN ANATOMIC
SITES AND ACKNOWLEDGING THE
LIMITATION OF OUR STUDY, WE
DIDN'T QUERY THE PAST
POPULATION.
WE'RE CURRENCY ABOUT MAKING
ABOUT OVER STATEMENTS HOW OUR
FINDINGS RELATES TO PAST BUT IN
THE PATIENT POPULATION THERE
SHOULD BE STUDIES DESIGNED TO
SPECIFICALLY EVALUATE WHETHER OR
NOT THERE IS PERSISTENCE OF mRNA
IN DIFFERENT ANATOMIC SITES AND
TO WHAT DEGREE THAT PERSISTENCE
IS ASSOCIATED WITH THE CLINICAL
MANIFESTATIONS OF PSAC.
-- PASC.
>> HOW LIKELY IS IT OTHER
RESPIRATORY VIRUSES LIKE COMMON
COLD HAVE WIDESPREAD TROPISM AND
PERSISTENCE IN NON-RESPIRATORY
TISSUES OR HAS IT GONE
UNDETECTED BUT THERE?
>> THAT'S A GREAT QUESTION AND
ONE WE HAVE HAD ENJOYABLE
CONVERSATIONS AND DEBATES AROUND
AND THE IMPLICATIONS ARE SHOULD
WE EXTEND THE APPROACH TO FILL
IN THE BLANK X, Y AND Z
RESPIRATORY OR OTHER VIRAL
ILLNESSES THAT ARE ASSOCIATED
WITH HUMAN SEVERE ILLNESS.
WHAT I'LL SAY IS THIS, THAT PART
OF OUR APPROACH FOR COV2 WAS
INFORMED BY THE HISTORIC
LITERATURE OF THE RECEPTOR MERS
DIFFERENT DIFFERENT RECEPTOR AND
SMALLER BODY OF LITERATURE BUT A
LITERATURE THAT FAIRLY DEFINES
THE CORONAVIRUSES AT LEAST THE
SEVERE HUMAN CORONAVIRUSES MAKE
THEIR WAY OUTSIDE THEIR
RESPIRATORY TRACT AND INFECT
TISSUES ELSEWHERE.
BASED ON THAT LITERATURE AND OUR
UNDERSTANDING, WE PURSUED THIS
QUOTE, UNQUOTE, AGGRESSIVE
APPROACH TO BETTER UNDERSTAND
COV2.
IF YOU LOOK AT THE LITERATURE
FOR EXAMPLE, INFLUENZA, I DON'T
THINK PEOPLE HAVE TAKEN THE
APPROACH WE HAVE TO LOOK AT
DISTRIBUTION ACROSS THE BODY AND
BRAIN BUT HAVING SAID THAT,
PEOPLE HAVE BEEN STUDYING
INFLUENZA FOR A LONG TIME AND
HAVE DONE A LOT OF CAREFUL
STUDIES AND THOSE STUDIES DO
SUGGEST THE PREVAILING KNOWLEDGE
AND DOGMA IS THE VIRUSES ABSENT
THE HIGH PATH INFLUENZA VIRUSES
WITH A CLEAVAGE SITE THAT ALLOW
THEM TO GET OUTSIDE THE
RESPIRATORY TRACT DON'T DO THIS
AND WE'RE DISCUSSING THOSE AMONG
OURSELVES.
>> AND THERE'S NONE OF THE
ANECDOTAL CLINICAL NEUROLOGICAL
EVIDENCE PEOPLE SAY THEY HAVE
WITH EVEN HAVING EXPERIENCE
COVID PERSONALLY THE BRAIN FOG,
THE VERTIGO, THESE THINGS
PERSISTED FOR MONTHS AFTER I
TESTED NEGATIVE FOR THE VIRUS
AND THIS YOU DON'T HEAR FROM THE
FLU OR COMMON COLD.
>> IT'S REALLY TOUGH.
SUPER TOUGH.
THERE'S LITERATURE OUT THERE IN
THE NON-RESPIRATORY
COMPLICATIONS OF FLU BUT MOST OF
THEM APPEAR TO BE NOT DUE TO
ORGANS BUT INDIRECT EFFECTS BUT
THE PASC IS A MAJOR ISSUE AND
NOW THERE'S STRONG DATA
SUPPORTING THAT THIS IS NOT JUST
A BIG CONSTELLATION OF THINGS.
IT IS RELATED TO PRIOR INFECTION
AND SO BEGINNING TO UNRAVEL THE
PATHOGENESIS OF THAT I THINK IS
AND OBVIOUSLY THE NIH I THINK
HAS DEVOTED OVER A BILLION
DOLLARS IN RESEARCH FUNDING TO
THAT PARTICULAR QUESTION.
>> AND CAN YOU COMMENT ON THE
RECENT STUDY IN NATURE WHERE
THEY FOLLOWED PATIENTS WITH F
MRI AND HAVE SEEN SHRINKAGE OF
GRAY MATTER?
>> I LOVE THAT STUDY.
THAT STUDY'S FASCINATING.
STRONGLY SUGGESTIVE, RIGHT?
IT'S REALLY THAT STUDY FROM THE
U.K. TOOK ADVANTAGE OF THESE
INDIVIDUALS SYSTEMATICALLY
FOLLOWED BY MRI PRE-COVID AND
WERE ABLE TO FOLLOW THE SAME
PEOPLE POST-INFECTION AND LOOK
AT FUNCTIONAL REGIONS AND THEY
FOUND A STRONG SIGNAL THAT
SOMETHING'S NOT RIGHT.
AND WHETHER THAT SOMETHING IS
NOT RIGHT PERSISTENCE OF mRNA
IN THE BRAIN IS IMPORTANT TO
FIGURE OUT.
>> SO IMPORTANT.
THERE'S A QUESTION ABOUT THE
SENSITIVITY OF YOUR ASSAYS WHEN
YOU HARVESTED THE VIRUS.
COULD IT BE YOUR MISSING OUT ON
VIRUS LEVELS BECAUSE YOUR
SENSITIVITY'S NOT THERE IN THE
EX VIVO ASSAYS?
>> THE ANSWER IS ABSOLUTELY YES.
THOUGH IT'S A SIMPLE ANSWER,
IT'S A HARD MESSAGE TO TRANSLATE
TO THE AVERAGE LISTENER.
WE K
WE KNOW CELL CULTURE THE GOLD
STANDARD WAY BUT THE WAY TO GET
AROUND THIS OR TO TRY TO
INCREASE SENSITIVITY TO FIND
REPLICATION COMPETENT VIRUS IN
SAMPLES THAT DIDN'T GROW IN CELL
CULTURE IS YOU CAN MANIPULATE
YOUR CELL CULTURE AND PUT IN
HUMANIZED RECEPTORS AND TAKE IT
TO A DIFFERENT SYSTEM IN OTHER
VIRUSES AS YOU ALL KNOW PEOPLE
HAVE USED IMMUNOCOMPROMISED MICE
THAT ARE NEONATAL MICE THAT
PERHAPS ARE PARTICULARLY
SUSCEPTIBLE.
THERE'S OTHER WAYS BUT VIRAL
CULTURE IS AN INSENSITIVE WAY OF
DETECTING THIS AND IS THERE
LIKELY THAN MORE OBSERVING
PROBABLY BUT WE HAVE TO WORK
WITH WHAT WE HAVE.
>> THERE'S A QUESTION FROM JOHN
SINER.
I WONDER IF YOU LOOK AT WHAT
PERCENTAGE OF PEOPLE YOU LOOKED
AT HAD PERSISTENCE IN THE BRAIN.
JI WAS CAREFUL AND I'LL
REITERATE TO ACKNOWLEDGE A
LIMITATION OF OUR STUDY IS THAT
THIS WAS DONE DURING THE FIRST
YEAR OF THE PANDEMIC BEFORE
AVAILABILITY OF VACCINES AND
NONE OF OUR COHORT WERE
VACCINATED.
WE KNOW, AS I KNOW THE PERSON
WHO ASKED THE QUESTION KNOWS,
THAT VACCINES ARE REMARKABLE AS
FAR AS PREVENTING SEVERE DISEASE
AND DEATH.
HOW GOOD ARE THEY AT PREVENTING
VIRAL DISSEMINATION TO EXTRA
PULMONARY SITES?
I DON'T KNOW AND WOULD THINK
THEY'RE PROBABLY PRETTY GOOD BUT
AGAIN A QUESTION WORTH ASKING.
WE KNOW THAT FOR EXAMPLE AND
AGAIN NOT TO CONFOUND ISSUES
HERE BUT EVEN VACCINATED PEOPLE
WHO HAVE BREAKTHROUGH INFECTIONS
THAT GO ON TO EXPERIENCE
SYMPTOMS OF PASC, MILD ILLNESS
YOU CAN HAVE IT AND
POST-BREAKTHROUGH INFECTIONS YOU
CAN HAVE IT.
I'M NOT TRYING TO CONFOUND THOSE
QUESTIONS BUT IDENTIFYING
QUESTIONS THAT REMAIN TO BE
ANSWERED AND THAT IS A QUESTION
THAT REMAINS TO BE ANSWERED.
>> HOWARD YOUNG IS ASKING IF
THERE WAS AN INTERFERON
SIGNATURE IN THE BRAIN?
>> HOWARD, YOU ASK A VERY GOOD
QUESTION AND THE ANSWER IS WE
DON'T KNOW YET.
THAT'S ON THE LIST OF FUTURE
WORK AND TO BE SPECIFIC THIS
FIRST EFFORT WAS REALLY FOCUSSED
ON THE PATHOGEN, AS YOU NOTICE.
OUR GROUP AND OUR NUMBER OF
COLLABORATORS BOTH WITHIN NIAID
AND MANY OTHER INTRAMURAL
INSTITUTES ARE BEGINNING TO
SHIFT OUR FOCUS TO APPLY VARIOUS
TECHNOLOGIES TO FOCUS ON MORE ON
THE HOST RESPONSE BEYOND
HISTOPATHOLOGIC EVIDENCE OR
EVIDENCE OF INFLAMMATORY CELLS
IN TISSUES.
AND ONE OF THOSE STUDIES IS
DIRECTLY FOCUSSED ON CELLULAR
RESPONSES IN THE BRAIN USING
AVAILABLE TISSUE.
ONE OF THE BENEFITS OF
COLLECTING FLASH FROZEN
SPECIMENS THAT ARE IMMEDIATELY
ADJACENT TO SPECIMENS WHERE
WE'VE CHARACTERIZED IN GREAT
DETAIL RNA LEVELS BY DDPCR IS
THOSE FLASH FROZEN SAMPLES ARE
AMENABLE TO TECHNOLOGIES LIKE
SINGLE NUCLIDE SEQ AND
APPLICATION OF ADDITIONAL
TECHNOLOGIES WHERE FOR EXAMPLE
SPATIAL TRANSCRIPTOMICS AND
THOSE THINGS TO BEGIN TO ANSWER
THAT TYPE OF QUESTION.
OTHERS HAVE APPLY THOSE TOOLS IN
THE BRAIN AND ELSEWHERE.
WE DO INTEND TO PURSUE THESE
THINGS IN OUR COHORT.
>> OKAY, WELL, THANK YOU SO
MUCH, DAN.
IT'S BEEN VERY ENLIGHTENING AND
THIS ENDS TODAY'S SEMINAR.
THANK YOU.