>>WELCOME BACK TO THE ASSAY
WORKSHOP. WE ARE EXCITED FOR YOU
TO BE HERE TODAY. WE HAD AN
AMAZING DAY YESTERDAY. WITH A
LOT OF DISCUSSIONS AND HOPEFULLY
YOU LEARNED A LOT AND IT WAS
BENEFICIAL FOR YOUR RESEARCH. WE
ARE LOOKING FORWARD TO AN
AMAZING DAY TWO TONS OF
DISCUSSIONS AND A LOT OF
LEARNING HAPPENING. SO I AM
EDITOR IN CHIEF AND ASSAY
GUIDANCE MANUAL AND LEAD OF THE
ASSAY GUIDANCE MANUAL
TRANSLATIONAL RESOURCES PROGRAM
HERE AT THE NATIONAL CENTER FOR
ADVANCING TRANSLATIONAL SCIENCES
AND I WILL BE HOSTING THE
WORKSHOP TODAY. LET ME SHARE
THE AGENDA FOR DAY TWO. WE WILL
HAVE AMAZING THREE SESSIONS
TODAY. LOOKING FORWARD TO THEM.
THE FIRST SESSION WILL BE ON
EWETY OF THE EXISTING 3 --
UTILITY OF 3D MODELS FOR
ANTIVIRAL DRUG DEVELOPMENT,
THAT'S SESSION 2. AND THEN IT
WILL START NOW AND SPAN ALL THE
WAY TO 2:30. WE WILL THEN HAVE
ANOTHER SESSION THIS AFTERNOON,
SESSION 3 WHICH WILL BE ON USING
ROBUST AND REPRODUCIBILITY 3D
MODELS THROUGH DRUG DISCOVERY
AND DEVELOPMENT. AND WE WILL
CONCLUDE WITH A CLOSING SESSION
ON SUMMARIES OF DISCUSSIONS AND
PERSPECTIVES ON THE CHALLENGES
AHEAD. SO STAY TUNED WITH US
AND ENJOY TODAY. AND BEFORE I
INTRODUCE CHAIR OF SESSION 1 I
WOULD LIKE TO REMIND EVERYONE
THAT YOU CAN ASK QUESTIONS
THROUGHOUT THE TALKS THROUGH THE
Q&A CHAT BOX AND WE WILL GET TO
THESE QUESTIONS AT THE END OF
THE EACH SESSION. THERE IS A
DISCUSSION AND Q&A TIME
DEDICATED TO ANSWER YOUR
QUESTIONS. AND WE GOT PLENTY OF
QUESTIONS YESTERDAY, HOPEFULLY
WE GET A LOT OF QUESTIONS TODAY
AND WE WILL COVER AS MANY
QUESTIONS AS POSSIBLE. SO WE
WILL START WITH SESSION 2 NOW,
AGAIN ENTITLED UTILITY OF
EXISTING 3D TISSUE MODELS FOR
ANTIVIRAL DRUG DEVELOPMENT. THIS
SESSION IS CHAIRED BY DR. ANN
EAKIEAKIEAKIN. DR. EAKIN IS SENR
SCIENTIFIC OFFICER, OFFICE OF
BIODEFENSE RESEARCH RESOURCES
AND TRANSLATIONAL RESEARCH IN
THE DIVISION OF MICROBIOLOGY AND
INFECTIOUS DISEASES OF THE
NATIONAL INSTITUTES OF ALLERGY
AND INFECTIOUS DISEASES HERE AT
THE NATIONAL INSTITUTES OF
HEALTH. THANK YOU SO MUCH, ANN
FOR BEING HERE. I WILL PASS THE
BATON TO YOU.
>> EXCELLENT. AGAIN TO ALL
PARTICIPANTS WELCOME BACK TO THE
MEETING. I THINK IT WAS A
FABULOUS DAY YESTERDAY,
EXCELLENT SPEAKERS, SET THE BAR
PRETTY HIGH FOR THE SPEAKERS
TODAY BUT I AM QUITE CONFIDENT
THAT WE CAN DELIVER. SO VERY
PLEASED TO LEAD THE SESSION 2
DISCUSSION. AGAIN ON THE UTILITY
OF -- UTILITY OF EXISTING 3D
TISSUE MODELS FOR ANTIVIRAL DRUG
DEVELOPMENT. WE HAVE SIX REALLY
FABULOUS SPEAKERS AND AGAIN
PLEASE DON'T HOLD BACK, WRITE
YOUR QUESTIONS IN AS THE TALKS
GO ALONG AND WE WILL HAVE PLENTY
OF TIME FOR Q&A AT THE END. IF
YOUR QUESTION IS DIRECTED I
WOULD LIKE TO ASK TO SPECIFIC
SPEAKER IF YOU CAN PUT THEIR
NAME IN THE FRONT THAT WILL HELP
ME COORDINATE THAT DISCUSSION,
IF IT IS TO ALL THE SPEAKERS
THAT IS GREAT. THAT CAN WORK
TOO. SO LET'S GO AHEAD AND KICK
OFF WITH OUR FIRST SPEAKER OF
THE SESSION. DR. DONALD INGBAR.
DR. INGBAR IS THE FOUNDING
DIRECTOR OF THE WYSS INSTITUTE
FOR BIOLOGICALLY INSPIRED
ENGINEERING AT HARVARD
UNIVERSITY. HE'S A FOLKMAN
PROFESSOR OF VASCULAR BIOLOGY AT
HARVARD MEDICAL SCHOOL AND
VASCULAR BIOLOGY PROGRAM AT
BOSTON CHILDREN'S HOSPITAL. AND
PROFESSOR OF BIOENGINEERING AT
THE HARVARD JOHN A PAULSON
SCHOOL OF ENGINEERING AND
APPLIED SCIENCES. HE RECEIVED
MULTIPLE DEGREES INCLUDING M.D.
AND Ph.D. FROM YALE
UNIVERSITY, DR. INGBAR IS
PIONEER IN THE FIELD OF
BIOLOGICALLY INSPIRED
ENGINEERING THAT CROSSES A RANGE
OF DISCIPLINES, TO REALLY FOCUS
ON DEVELOPING BREAK THROUGH
TECHNOLOGIES TO ADVANCE
HEALTHCARE. HE'S AUTHORED MORE
THAN 500 PUBLICATIONS AND ALMOST
200 U.S. PATENTS. HE'S FOUNDED
SEVEN COMPANIES AND MADE GREAT
STRIDES IN TRANSLATING THE
SCIENTIFIC DISCOVERIES AND
INNOVATIONS HIS TEAM HAS MADE
INTO COMMERCIAL PRODUCTS, MANY
OF WHICH ARE NOW EITHER IN
CLINICAL TRIALS OR
COMMERCIALIZED. DR. INGBAR IS
MEMBER OF THE NATIONAL ACADEMY
OF MEDICINE, CASHNAL ACADEMY OF
-- NATIONAL ACADEMY OF
ENGINEERING, AMERICAN INSTITUTE
FOR AMERICAN BIOLOGICAL
ENGINEERING AND THE AMERICAN
ACADEMY OF ARTS AND SCIENCES. HE
IS ALSO LISTED AMONG NATURE'S
BIOTECHNOLOGIES TOP 20
TRANSLATIONAL RESEARCHERS IN
2012, 2019 AND 2020. SO REALLY
PLEASED TO HAVE DR. INGBAR KICK
OFF THIS SESSION. AND THE TITLE
OF HIS TALK IS MODELING
RESPIRATORY VIRAL INFECTIONS IN
HUMAN LUNG CHIPS OVER TO YOU.
>> THANK YOU SO MUCH. CAN YOU
SEE MY SCREEN IN
>> SURE CAN.
>> GREAT. THANK YOU FOR THE
OPPORTUNITY. WHAT I WOULD LIKE
TO DO IS DESCRIBE TO YOU DRUG
DISCOVERY THAT WE HAVE BEEN ABLE
TO CARRY OUT OVER THE LAST
COUPLE OF YEARS, IT'S BEEN
ENABLED BY MODELING RESPIRATORY
VIRUS INFECTIONS USING HUMAN
LUNG CHIPS. WE HAVE WORKED FOR
15 YEARS NOW CALLED HUMAN ORGAN
ON CHIPS OR ORGAN CHIPS. THESE
ARE MICRODEVICES CONTAINING
LIVING HUMAN CELLS THAT
RECONSTITUTE ORGAN LEVEL
STRUCTURE AND FUNCTIONS NOT
CELLULAR TISSUE BUT ORGAN LEVEL
FUNCTION THAT ARE CREATED WITH
ORIGINALLY WITH MICROCHIP
MANUFACTURING TECHNIQUES, CALLED
ORGAN CHIPS. WE DEVELOPED THEM
TO ACCELERATE DRUG DEVELOPMENT,
REPLACE ANIMAL TESTING AND
REPLACE PERSONALIZED MEDICINE.
OUR FIRST CHIP WAS A LUNG CHIP,
WE CALLED IT HUMAN BREATHING
LUNG CHIP, IT IS A MODEL OF THE
LUNG ALVEOLUS AND I WANT TO
EMPHASIZE WE ARE T NO
ENGINEERING THE WHOLE LUNG, THIS
IS ARE LIVING THREE DIMENSIONAL
CROSS SECTIONS FOR MAJOR
FUNCTIONAL UNIT THE ALVEOLUS.
THE NEXT VIDEO SHOWS HOW THIS
WORKS. IT IS SIZE OF COMPUTER
MEMORY STICK SHOWN AT THE TOP
RIGHT. MADE OUT OF OPTICALLY
CLEAR FLEXIBLE RUBBE RUBBER. IFU
CUT IN CROSS SECTION THREE
HOLLOW CHANNELS EACH LESS THAN A
MILLIMETER WIDE. THE MIDDLE ONE
IS CUT INTO TOP AND BOTTOM WITH
A POROUS MEMBRANE WITH EXTRA
CELLULAR MATRIX. SO WE TAKE
ALVEOLAR CELLS HUMAN CELLS ON
TOP, HUMAN LUNG CAPILLARY CELLS
ON THE BOTTOM, WE RECREATED THE
ALVEOLAR CAPILLARY INTERFACE.
TRICK IS THAT WE HAVE SIDE
CHAMBERS THAT ARE HOLLOW WE
APPLY SUCTION, EVERYTHING IS
FLEXIBLE AND ACTUALLY STRETCHES
IN THE SAME RATE AND DEGREE AS
WHEN WE BREATHE IN AND OUT. WE
CAN NOW FLOAT AIR OVER THE TOP
TO HAVE AIR LIQUID INTERFACE,
PUT CAPILLARY CELLS ON ALL FOUR
SIDES AND FLOW MEDIUM WITH
IMMUNE CELLS OR WHOLE BLOOD
WITHOUT ANTI-COAGULANTS FOR
SHORT TIMES. IF THIS WORKS MODEL
ORGAN LEVEL RESPONSE IMAGINE YOU
HAVE A BACTERIAL INFECTION OR
VIRAL INFECTION THIS USUALLY IS
A TISSUE TISSUE SIGNALING
RESPONSE, SO EPITHELIAL CELLS
PUT OUT CYTOKINES ACTIVATE
ENDOTHELIUM, WHITE BLOOD CELLS
THAT WERE JUST FALLING BEHIND
STICK AND ROLL, MIGRATE ACROSS
AND ENGULF THE BACTERIA. THAT'S
BECAUSE THE CYTOKINES INDUCE THE
ENDOTHREE LYNN TO PUT OUT
ADHESION RECEPTORS. THESE ARE
FRESH WHITE BLOOD CELLS LABELED
FLUORESCENTLY, YOU DON'T SEE THE
ENDOTHELIUM, AND THE EPITHELIUM
IS BEHIND THE SCREEN YOU CANNOT
SEE. TO BEGIN WITH QUIESCENT,
THEY FLOW BY, BACTERIA ON THE
OTHER SIDE THERE IS SIGNALING
RESPONSE, I CAM IS ON THE OTHER,
THEY ARE PULLED OUT UNDER FLOW
KNOWN THE INITIAL ADHESION IS
SHEER STRESS DEPENDENT. AND NOW
YOU CAN DO ANY IMAGING IN THE
DEVICE YOU CAN DO OTHERWISE HERE
IS ONE WHITE BLOOD CELL HERE
BETWEEN TWO END HEELIAL CELLS,
FINDS MATRIX PORE, MIGRATE OUT
OF FOCUS TO THE OTHER SIDE, NOW
YOU CAN SEE IT COMING OUT OF THE
EPITHELIUM BY PHASE CONTRAST.
NOW SHOW YOU THE WHITE BLOOD
CELL IN RED, THE BACTERIA ARE
LABELED IN GFP GREEN, WATCH THEM
BEING ENGULFED SO YOU WATCHED
THE ENTIRE HUMAN INFLAMMATORY
RESPONSE IN THIS CHIP. MULTIPLE
PUBLICATIONS WITH THIS CHIP BUT
BASICALLY DEMONSTRATED PROOF OF
PRINCIPLE FOR DISEASE MODELS, WE
HAVE DONE PULMONARY EDEMA, THIS
DRUG TOXICITY EVEN NOT SEEN IN
ANIMALS, DRUG EFFICACY, WE
DISCOVERED THERAPEUTIC TARGETS,
AS A DRUG PHASE 2 CLINICAL
TRIALS CAME OUT OF THE THIS WORK
IN PART AND WE HAVE DELIVERED
GENE THERAPIES ADENOVIRAL
VECTORS ON THESE CHIPS. AND IN
ADDITION TO IMMUNE CELLS WE HAVE
INTEGRATED STROMAL FIBROBLASTS
SO YOU CAN LOOK AT PULMONARY
FIBROSIS WITH FUNDING FROM THE
FDA. MULTIPLE AWARDS I WON'T GO
INTO IN THIS TALK BUT IT REALLY
REPRESENTED A PARADIGM SHIFT IN
THE FIELD I THINK THAT PEOPLE
RECOGNIZE THIS IS POSSIBLE. SO
WE BUILD SOMETHING LIKE 20
DIFFERENT ORGAN CHIPS BUT THE
SECOND WE BUILT WAS A LUNG
AIRWAY CHIP BECAUSE THE
PHARMACEUTICAL COMPANIES WERE
INTERESTED IN ASTHMA AND COPD,
AND HERE WE TAKE PRIMARY HUMAN
LUNG BRONCHIAL EPIHELIUM, HUMAN
LUNG ENDOTHELIUM AND CULTURE IN
AIR LIQUID INTERFACE AND NOW YOU
HAVE INCREDIBLE BEAUTIFUL
DIFFERENTIADIFFERENTIATION. YOUT
FLUORESCENT PARTICLES AT THE
RIGHT, YOU LOOK AT MUCOCILIARE
CLEARANCE, THE SAME VELOCITY AS
RO1 AT THE MOMENT, ALL THE CELL
TYPES IN NORMAL RATIOS YOU GET
NORMAL EPITHELIAL MORPHOLOGY. WE
MADE CHIPS WITH CELLS FROM
PATIENTS WITH DISEASES IN THIS
CASE CHRONIC OBSTRUCTIVE
PULMONARY DISEASE, COPD. WHAT
BRINGS THE PATIENTS TO THE
HOSPITAL USUALLY EXACERBATIONS
BY BACTERIAL INFECTION VIRAL
INFECTION OR CIGARETTE SMOKE AND
THE FIRST THING WAS THAT WE CAN
SHOW THAT YOU RETAIN THE THE
COPD PHENOTYPE, NO IMMUNE CELLS
HERE, MUST BE EPIGENETIC CHANGES
IN THE EPITHELIUM IN TERMS OF
DECREASE TOLL 3 R 4 RECEPTOR
EXPRESSION. THEN IF YOU USE
POLYVIRAL INFECTION OR LPS
ENDOTOXIN ONLY THE COPD SHOW
INCREASE RESPONSE TERMS OF IL 8
OR MCSF SECRETION. WE BUILT A
CIGARETTE SMOKING ROBOT THAT HAS
LIGHTER FROM CAR AND REAL
CIGARETTES MIMICKING SMOKING
PARAMETERS REAL SMOKE NOT
EXTRACT GOES TO AIR SPACE, AND
JUST TO SHOW YOU COUPLE OF
EXAMPLES LOOKING AT IL 8 YOU
HAVE HEALTHY CHIPS CIGARETTE
SMOKE EXPOSURE DOES NOTHING. IN
COPD CHIPS IT DOUBLES. THE RIGHT
YOU SEE TRANSCRIPTOMIC RESULTS
WE DO -- WE HAVE DONE PROTEOMIC
METABOLOMIC GLYCOMIC AND
TRANSGENOMIC CHIPS ON THE RIGHT
THEY ARE HEALTHY CHIPS FROM
HEALTHY PATIENTS AFTER CIGARETTE
SMOKE EXPOSURE. AT THE LEFT IS A
PAST CLINICAL STUDY WITH HEALTHY
PEOPLE WHO SMOKE CIGARETTES.
WHAT YOU CAN SEE IS THAT THE TOP
QUARTER VERSUS BOTTOM THREE
QUARTERS ARE SIMILAR, WE GOT
PUSH BACK BECAUSE OF VARIATION
IN OUR CHIPS WAS LESS THAN THE
PATIENT POPULATION. THEN WE
REALIZE WE CAN DO MATCH
COMPARATIVE MODELING IN THESE
CHIPS MEANING A BETTER CLINICAL
STUDY IN THE SENSE THAN REAL
CLINICAL STUDY BECAUSE THESE
GENES ARE DUE TO CIGARETTE SMOKE
NOT FAMILY HISTORY, WORK
ENVIRONMENT, ET CETERA. THIS
WAS PUBLISHED WITH A LITTLE
COMMENTARY EXPLAINING THAT. TO
GET TO THE POINT OF TODAY'S
TALK, WHEN WE STARTED THIS THE
IDEA WAS TO AXEL INVESTIGATE
DRUG DEVELOPMENT BY REPLACING
ANIMAL TEST E. CXFCG. CXFC BEWE
REALIZED THESE CHIPS PROVIDE A
WINDOW ON MOLECULAR SCALE
ACTIVITIES INSIDE THE LIVING
CELLS WITHIN A RELEVANT TISSUE
AND ORGAN LEVEL CONTEXT. AND
THAT'S THEY POTENTIALLY ENABLE
MECHANISTIC INSIGHTS AND DRUG
DISCOVERY AND AT THE SAME TIME
PROVIDE EXPERIMENTALLY TEST
PREDICTIONS WE HAVE, ALL THE
HUMAN RELEVANT MODEL. ABOUT
2017, WE WERE FUNDED BY DARPA
AND NIH TO LEVERAGE OUR AIRWAY
CHIPS TO DEVELOP MODELS OF VIRAL
INFECTION AND THERE WAS GREAT
INTEREST AT THAT TIME IN
POTENTIAL PANDEMIC RESPIRATORY
VIRUSES BUT THE FOCUS WAS ON
INFLUENZA. SO WE TOOK OUR AIRWAY
CHIP FOR EXAMPLE, THIS IS GFP
LABELED H 1N 1 INFLUENZA AND YOU
CAN WATCH CELLS INFECTED REAL
TYPE ON THE CHIP. YOU CAN YOU
HAVE BEAUTIFUL MORPHOLOGICAL
READ OUTS OF INFECTION GFP VIRUS
AT THE BOTTOM LEFT YOU LOSE
JUNCTIONS, CILIA, YOU CAN SEE
ENDOTHELIAL DAMAGE. WE DID
DIFFERENT VIRAL STRAINS AND YOU
CAN SEE VIRAL STRAIN DEPENDENT
EFFECTS IN TERMS OF REPLICATION
AND DIFFERENT VIRULENCE IS
MIMICKED IN THESE CHIPS, THE
SAME DIFFERENCES VIRULENCE YOU
SEE IN PATIENTS BETWEEN H 1N 1
AND H 5N 1, WE CAN BEGIN TO
EXPLORE EFFECTS OF CO-MORBIDITY.
SO WE DID THE SAME STUDY WITH
COPD CHIPS AND SEE TENFOLD
HIGHER INCREASE IN VIRAL
REPLICATION COPD PATIENTS ARE
MORE SENSITIVE TO INFLUENZA
INFECTION. PERHAPS MOST
IMPORTANTLY WE CAN MEASURE HOST
RESPONSE TO INFECTION AND DRUGS
IN TERMS OF WHAT IS A BASICALLY
A CYTOKINE STORM USING COVID
TERMS, THESE ARE IL 6 IP 10 AND
FOUR OTHER CYTOKINES IN RESPONSE
TO THREE TYPES OF INFLUENZA AND
ONCE AGAIN THE CYTOKINE RESPONSE
MATCHES THE DEGREE OF VIRULENCE
SEEN IN PATIENTS. WE CAN DO
IMMUNE CELL RECRUITMENT AND IN
THIS CASE WE WATCHED IMMUNE
CELLS MIGRATE INTO THE
EPITHELIUM AND CLEAR THE VIRALLY
INFECTED CELLS OVER COUPLE OF
DAYS. THEN WE TESTED DRUGS SO
TAMI FLU, THE MOST COMMON IS
ANTI-INFLUENZA DRUG. WE SEE
BEAUTIFUL IN ADDITION ON THE
CHIPS, OVER THE SAME TWO DAYTIME
COURSE THE FDA HAS APPROVED THIS
DRUG IN PATIENTS. INTERESTINGLY
WE ALSO CAN MODEL EVOLUTION OF
VARIANTS OF CONCERN IF YOU LIKE
ON THESE CHIPS. THIS STUDY WE
TOOK A HUMAN AIRWAY CHIP WE
INFECTED IT WITH INFLUENZA, AND
WE GAVE IT A DRUG AT 90%
EFFICACY LEVEL. IN THIS CASE WE
DID AMANTADINE AND AFTER COUPLE
OF DAYS WASHED THE DROPLETS AND
PASSED TO AIR SPACE OF ANOTHER
CHIP LIKE SOMEBODY WAS COUGHING
HUMAN TO HUMAN TRANSMISSION. WE
DID IT AGAIN AND AGAIN. WITH
IMANTADINE IN EIGHT PASSAGES WE
GOT RESISTANT VIRAL VAINS WHEN
WE DID GENOME SEQUENCING WE
IDENTIFIED ALL KNOWN MUTATIONS
PUBLISHED IN THE LITERATURE
BASED ON CLINICAL EXAMPLES
PATIENTS BECAME RESISTANT.
CLOSELY IDENTIFIED MULTIPLE
MUTATIONS NEVER SEEN BEFORE. SO
THIS COULD IDENTIFY VARIANTS OF
CONCERN COMING DOWN THE PIKE FOR
VACCINE AND THERAPEUTIC
DEVELOPMENT. WE DID THIS WITH
TAMI FLU, 25 OR 28 PASSAGES WE
ALSO GOT RESISTANT VIRAL MUTANTS
WHICH WAS TERRIFYING. NOW, THE
REASON I SHOWED YOU THE COPD
PAPER, THIS LAUNCHED US TO BEGIN
TO LOOK AT DRAPERY PURPOSING FOR
VIRAL -- DRUG REPURPOSES FOR
VIRAL INFECTION. WE NOTICE
UPREGULATION OF MULTIPLE SERENE
PROTEASES AND ABOUT THESE ARE
HOST SERENE PROTEASES TO TARGET
HOST RATHER THAN VIRUS AND
THERE'S BEEN SOME PAST WORK TO
SUGGEST THAT SO WE TOOK TWO
EXISTING DRUGS, ANTI-COAGULANT
AND TRAZAWOL BOTH GENERAL
PROTEASE INHIBITORS AND WE HAVE
BEAUTIFUL INFECTION OF H 1N 1 H
3N 2 BY BOTH OF THESE ON THE
CHIPS. BUT WHAT WAS REALLY
EXCITING IS THAT WE CAN TAKE
NAFAMOSTAT AND COMBINE IT WITH
TAMIFLU AND DOUBLE TAMI FLU
THERAPEUTIC WINDOW FROM 48 TO 96
HOURS WHICH COULD HAVE REAL
CLINICAL RELEVANCE GIVEN TAMI
FLU IS ONLY APPROVED FOR 48
HOURS. THIS IS WHEN COVID HIT
END OF JANUARY 2020. I HAVE TWO
VIROLOGISTS POST DOCS WHO ARE
FROM CHINA FOLLOWING THIS ON
SOCIAL MEDIA. WITHIN ONE DAY
AFTER THE PUBLICATION OF THE
GENOME SEQUENCING SARS COV-2 IN
SCIENCE, THEY ENGINEERED A SARS
COV-2 SPIKE PROTEIN EXPRESSING
PSEUDOTYPE VIRUS BECAUSE WE ONLY
HAVE A BSL 2 LAB. THEY ALSO HAD
BEEN SEARCHING THROUGH THE
LITERATURE TO QUICKLY MAYBE TAKE
EXISTING APPROVED DRUG TO USE
FOR THIS INCREDIBLE CLONALITY
HAPPENING AT THAT TIME AND THEY
IDENTIFIED 30 DRUGS SHOWN
REPORTED ACTIVITIES AGAINST
EBOLA, MRS OR SARS COVE 1 AND
TESTED THEM IN PA CELL LINE THEY
DID IN GRADUATE WORK THAT
VIROLOGISTS USE LIVER CELL LINE
ACTUALLY. AND YOU CAN SEE THE
EIGHT DRUGS THAT SHOWED DOSE
DEPENDENT INHIBITION IN THE LOW
MICROMOLAR RANGE. THIS IS
FEBRUARY OF 2020.
HYDROXYCHLOROQUINE AND COLOR
QUINN AMONG THESE EIGHT. WE THEN
THOUGHT WE SHOULD TEST THESE IN
OUR HUMAN LUNG AIRWAY CHIPS. I
SHOULD NOTE H 287 OFTEN USED BY
VIROLOGISTS HAVE NORMAL
INTERFERON VIRUSES THAT'S WHY
VIRUSES GROW WELL, THEY DON'T
HAVE THE RIGHT RECEPTORS FOR
CERTAIN VIRUSES LIKE SARS COV-2
SO WE CONFIRMED THAT THE HUMAN
LUNG AIRWAY CELLS WHEN
DIFFERENTIATE HAVE HIGH LEVELS
OF ACE 2 AND TMPRSS 2, PROTEASE
AND RECEPTOR CRITICAL FOR COV 2
INFECTION. WE CONFIRMED OUR
PSEUDOTYPE VIRUS BINDS TO CILIA
WHICH JUST WAS REPORTED IN VIVO
IN LUNGS AS WELL. THIS IS ON OUR
CHIPS TOP VIEW AND CROSS
SECTIONAL VIEW. AND NOW WE
INSTEAD OF BATHING THE CELL IN
DRUG WE FLOW THE DRUG THROUGH AT
CLINICALLY RELEVANT DOSE CMAX
THROUGH THE CHIP. WE FIND
HYDROXYCHLOROQUINE HAVE NO
EFFECT NOR A NEW OTHER DRUGS
TESTED IN CLINIC THAT WERE NOT
EFFECTIVE. AND WE FOUND A COUPLE
OF DRUGS THAT SHOWED MORE POTENT
ACTIVITY WITH AMADAQUIN ANOTHER
ANTI-MALARIA IN AFRICA FOR YEARS
IN PARTICULAR KIDS WITH
PROPHYLAXIS FOR MALARIA WAS
SIGNIFICANTLY EFFECTI EFFECT EFE
DATA I HAVE BEEN COMMUNICATING
WITH PEOPLE AT GATES AND DARPA
AND THIS TRIGGERED US TO GET
FUNDING FROM DARPA TO DEVELOP A
LARGER DRUG REPURPOSING PROGRAM
THAT NOT ONLY LEVERAGE WHAT WE
HAD DONE THERE BUT OTHER
TECHNOLOGIES WE DEVELOPED IN MY
LAB FOR DRUG REPURPOSING, ONE
USING MOLECULAR DYNAMIC
SIMULATION AN STRUCTURAL
SIMILARITY AND STRUCTURE
ACTIVITY RELATIONSHIPS WITH
MEDICINAL CHEMISTRY. THE OTHER
IS MACHINE LEARNING AND SORT OF
NETWORK ANALYSIS COMPUTATIONAL
PATHWAY CALLED NEMOCUT THAT
STARTS WITH TRANSCRIPTOMIC DATA
FOR PATIENTS, I WILL GO INTO
MORE. THEY LINK ME --
COLLABORATED WITH MATT FREEMAN,
UNIVERSITY OF MARYLAND EXPERT IN
CORONA VIRUS AND HAS BSL 3 AND
TENANT OVER AT MOUNT SINAI WHO
DEVELOPED HAMSTER SARS COV-2
MODELS. THE FIRST THING WE DID
IS CONFIRM THAT AMODAAQUIN DOES
AFFECT POTENTLY NATIVE SARS
COV-2, METABOLITE HAS ACTIVITY.
BEEN CONFIRMED THAT WE HAVE
GREATER ACTIVITY IN HUMAN 8549
CELLS EXPRESSING ACE 2 BUT THEN
HE TESTED THE DRUG IN HAMSTER
MODEL AND WHEN YOU CAN SEE HERE
IS NOT HYDROXY COLOR QUINN,
INHIBITS INFECTION BY SARS COV-2
AND ADDED ONE DAY BEFORE
INFECTION PROPHYLACTICICALLY
POTENT INHIBITION FROM ANIMAL TO
MAN MALL TRANSMISSION AND
INHIBITS WHEN YOU START
TREATMENT AFTERWARDS. IN PART
THIS DATA THIS DRUG IS NOW IN
CLINICAL TRIALS ACROSS 20 SITES
IN AFRICA FOR COVID-19. MORE
RECENTLY, WE DID WORK THAT WAS
MORE MECHANISTIC FOCUSED, WE
WERE LOOKING AT H 3N 2 INFLUENZA
INFECTION BECAUSE WE FOUND THAT
WAS A BETTER MODEL OF PNEUMONIA
IF YOU LIKE AND CLOSER TO COV 2
THAN COVID-19 THAN OTHER
UNRELATED COLD VIRUSES THAT ARE
CORONA VIRUS. AND MY POST DOC
NOTICED DID EXPERIMENT DID WITH
AND WITHOUT BREATHING MOTIONS IN
THE ALVEOLUS CHIP AND HE FOUND
THAT BREATHING SUPPRESSED VIRAL
INFECTION DRAMATICALLY,
PHYSIOLOGICAL BREATHING MOTIONS
AND SUPPRESSED CYTOKINE
INFLAMMATORY RESPONSE BASICALLY
STIMULATED THE INNATE IMMUNE
RESPONSE. WE DID SINGLE CELL RNA
-- I'M SORRY, WE DID BOTH RNA
SEQ AND HE NOTICED THERE WAS ONE
GENE THAT CAME UP WITH BREATHING
MORE THAN ANY OTHERS CALLED S
100A 7 WHICH IS A LIGAND FOR THE
RAGE RECEPTOR, THE RAGE WHICH IS
THE RECEPTOR FOR ADVANCE
GLYCATION PRODUCTS WHICH IS A
MAJOR PLAYER IN INFLAMMATION,
MOST HIGHLY EXPRESSED IN
ALVEOLAR CELLS. SO WE RETAINED A
DRUG A RAGE INHIBITOR AND
INHIBITED THE CYTOKINE STORM IF
YOU LIKE INDUCED BY VIRAL
INFECTION IN THESE CHIPS. AND IT
WAS IN PHASE 3 CLINICAL TRIALS,
2000 PATIENTS COMPLETELY SAFE
ORAL DRUG HIGHLY BIOAVAILABLE,
IT WAS IN CLINICAL TRIALS FOR
ALZHEIMER'S WHICH DID NOT HIT
THE MARK BUT BASED ON THESE DATA
AND EXTENSIVE LITERATURE
SUPPORT, WE LICENSED THIS IP
DISCOVERY TO CTEX PHARMA, THESE
VERY DATA WERE INCLUDED IN IND
APPLICATION INVATIONAL NEW DRUG
APPLICATION TO THE FDA TO START
COVID-19 TRIALS WHICH IS UNDER
REVIEW RIGHT NOW. I MENTIONED
THESE OTHER COMPUTATIONAL, WE
USED A COMPUTATIONAL PIPELINE,
WE DEVELOPED AT THE WYSS
INSTITUTE TO STUDY THE SPIKE
PROTEIN AND WHEN WE FOUND IS
THAT IT HAS TO UNDERGO MAJOR
OPENING SHOWN AT THE BOTTOM
RIGHT IN THIS MOVIE, TO BIND TH
2 -- MEDIATE FUSION WHEN IT
BINDS TO ACE 2 RECEPTOR AND WE
WERE ABLE TO IDENTIFY A
CONSERVED SITE INSIDE THE
MOLECULE THAT IS NOT INNATE SITE
WHERE ANTIBODIES HIT THAT IS
ALSO IN MANY OTHER CORONA VIRUS,
ALL OTHER CORONA VIRUS THAT WE
FELT IF WE CAN DESIGN MOLECULE
TO HIT THAT SITE WE MIGHT BE
ABLE TO INHIBIT INITIAL INFEC
INFECTION. WE USE THAT MODEL TO
LOOK LIEU EXISTING APPROVED
DRUGS TO SEE IF THERE'S ANY THAT
BIND THAT SITE AND WE IDENTIFIED
A COUPLE AND WE CAN SHOW THAT
THEY INHIBIT THE SPIKE PROTEIN
COV 2 SPIKE PROTEIN ENTRY. WE
USE MEDICINAL CHEMISTRY TO
DEVELOP NEWER VERSIONS OF THIS
THAT WE HAVE SYNTHETIC ANALOG
THAT ARE EVEN A LITTLE BIT MORE
POTENT. THEN WE HAVE OPTIMIZED
WITH FUNDING FROM NIH DARPA AND
OPEN PHILANTHROPY AND WE NOW
HAVE SYNTHETIC COMPOUNDS THAT
INHIBIT ALL OF THE PSEUDOTYPE
VIRUSES OF ALPHA BETA DELTA
GAMMA OMICRON ALSO SARS COV 1
AND MRSS, IT WORKS AGAINST
NATIVE COV 2 IN BSL 3 LAB WITH
150 NANOMOLAR IC 50 AND GREAT
BIOAVAILABILITY SO WE ARE NOW
MOVING THIS INTO ANIMAL EFFICACY
STUDIES WITH THE HOPE OF HAVING
ORAL PROPHYLACTIC IF YOU LIKE.
THE OTHER COMPUTATIONAL NETWORK
ANALYSIS PREDICTION I
MENTIONEDDED STARTS WITH
TRANSCRIPTOMES FROM PATIENTS
WITH COVID VERSUS HEALTHY. AND
BASICALLY TRIES TO SEE WHAT
GENES AND WHAT NETWORK
TOPOLOGIES OR NETWORK
RELATIONSHIPS HAVE TO CHANGE TO
MAKE A DISEASE PHENOTYPE OR
STATE SWITCH TO HEALTHY STATE.
AND THEN IT LOOKS FOR DRUGS THAT
HAVE KNOWN TRANSCRIPTIONAL
SIGNATURES PUBLISHED IN
DATABASES, GOES THROUGH THEM,
AND ESSENTIALLY SEARCHES LIEU
FOR DRUGS THAT ARE HIGHLY LIKELY
TO REVERSE THAT STATE. WE FOUND
DRUGS, THEY SHOWED EFFICACY
AGAINST SARS COV-2 IN IN VITRO
MODELS BUT ONE THING THAT CAME
UP WAS THAT IT PREDICTED
MULTIPLE STATINS WHICH IS LIKE
ONE OF THE MOST POPULARLY USED
DRUG AND SAFE DRUG, MULTIPLE
STATINS WERE PREDICTED TO HAVE
COVID REVERSING OR PROTECTING
ACTIVITY, BUT THEY NOT ALL
STATINS WERE THE SAME SUGGESTING
THIS MIGHT NOT BE RELATED TO ITS
KNOWN LIPID METABOLISM ACTIVITY.
AND WE ALSO FOUND FOR EXAMPLE
STATIN CAME UP AGAIN AND AGAIN.
SIMVA STATIN DEMONSTRATED DIRECT
INHIBITION OF SARS COV-2
INFECTION IN VITRO AND EVEN
INACTIVE METABOLITE HAD
ACTIVITY. SO WE ARE UNCOVERING
WE DO REPURPOSING ACTIVITIES
THAT ARE NOT THE KNOWN TARGETS
AND THIS HAS HAPPENED AGAIN AND
AGAIN. NOW, RECENTLY WE
COLLABORATED WITH MARIA SIROPTA
AND OTHERS AT STANFORD WHO
ANALYZED OVER 4,000 PATIENTS SO
WE DID A RETROSPECTIVE ANALYSIS,
INDEED WE CONFIRM THAT THERE ARE
SIGNIFICANT DIFFERENCES BETWEEN
DIFFERENT STATINS IF YOU LOOK IN
RED HERE, TORVA STATIN AND SIMVA
STATIN, DO PROTECT BUT OTHERS DO
NOT. SO AGAIN THIS WAS
ORIGINALLY COVERED BETWEEN
COMPUTATIONAL APPROACH AND OUR
IN VITRO MODELS. LASTLY, IN THE
COURSE OF WORKING ON THE
INTERFERON INFECTION WE CARRIED
OUT STUDIES WITH A CRISPR SCREEN
AND WE WERE ABLE TO IDENTIFY
SHORT DUPLEX RNAs, ORIGINAL
LIKING FOR LONG NON-CODING
RNAs THAT ARE POTENT INDUCERS
OF TYPE 1 INTERFERON LIKE
INTERFERON BETHAT ARE PROTECTIVE
INTERFERONS. AND FURTHER
ANALYSIS REALIZE THAT WE WEREN'T
WORKING THROUGH LONG NON-CODING
RNAs, IT WAS ACTUALLY THE
DUPLEX RNAs THEMSELVES HAD
UNIQUE SEQUENCES THAT MADE THEM
BASICALLY THE INTERFERON
INDUCERS. HERE YOU CAN SEE
INTERFERON BETA IN OUR CHIPS, WE
CAN SHOW WITH THIS RNA 1, THIS
IS 20 BASE PAIRS, VERY CHEAP TO
SYNTHESIZE COMPLETELY, YOU HAVE
30 FOLD INCREASE AND CLOSE TO 15
FOLD INCREASE IN AIRWAY ALVEOLAR
EPITHELIUM BUT MORE IMPORTANTLY,
THIS ONE MOLECULE CAN BE USED TO
INHIBIT INFLUENZA, H 1N 1 IN
CHIPS, H 3N 2 INFLUENZA, SARS
COV-2 WITH 4 LOG REDUCTION. MERS
AND THE COMMON COLD VIRUS IN L
63. WE HAVE CONFIRMED IN VIVO IN
SARS COV-2 MOUSE MODEL, WE GET 3
LOG REDUCTION USING THIS RNA,
THIS IS ACTUALLY INTRAVENOUS
INJECTION AND WE HAVE
PRELIMINARY DATA WITH
INTRATRACHEAL IN HAMSTERS BUT
DELIVERY SYSTEM WASN'T OPTIMIZED
BUT THIS LOOKS REALLY BEAUTIFUL.
SO TO END, I THINK WHAT I WOULD
LIKE TO COMMUNICATE IS THAT
THESE HUMAN ORGAN CHIPS ARE MORE
THAN POTENTIAL ANIMAL
REPLACEMENTS AND THEY ARE AT A
DIFFERENT LEVEL THAN ORGANOIDS.
THEY MECHANISTIC DISCOVERY DRUG
DISCOVERY TOOLS THAT PROVIDE NEW
INSIGHT INTO PATHOPHYSIOLOGY
THAT CAN BE COMBINED WITH
ADVANCED ANALYTICAL TECHNIQUES.
THEY CAN ENABLE RAPID DRUG
REPURPOSING AND ACCELERATE
DISCOVERY OF NOVEL THERAPEUTICS
AS WELL. WITH THAT I HAVE TO
DISCLOSE THAT I'M A MEMBER OF
THE BOARD, CHAIR OF THE SAB AND
EMULATE CELLS THESE CHIPS AND
AUTOMATED INSTRUMENTS WORLDWIDE.
AND I COULD NOT DO THIS WITHOUT
INCREDIBLY INTERDISCIPLINARY
TEAM AT THE WYSS INSTITUTE.
PLEASE VISIT THE WEBSITE IF YOU
WANT TO LEARN MORE. THANK YOU SO
MUCH.
>> THANK YOU, FASCINATING
PRESENTATION. WE ARE COLLECTING
QUESTIONS AND WILL HAVE
DISCUSSION THIS AFTERNOON.
SHIFTING GEARS NOW TO THE SECOND
SPEAKER IN THE SESSION, LIKE TO
INTRODUCE DR. LEE GEHRKE. DR.
GEHRKE IS A HERMAN LF VON
PROFESSOR OF HEALTH SCIENCES AND
TECHNOLOGY AT THE MASSACHUSETTS
INSTITUTE OF TECHNOLOGY. ALSO
PROFESSOR OF MICROBIOLOGY AT
HARVARD MEDICAL SCHOOL. DR.
GEHRKE WAS BORN IN RAISED IN
RURAL ILLINOIS BEFORE MOVING TO
CLEVELAND, OHIO, TO ATTEND CASE
WESTERN RESERVE UNIVERSITY FOR
GRADUATE SCHOOL. FOLLOWING THAT
HE DID A POST-DOCTORAL
FELLOWSHIP AT MIT. DR. GEHRKE'S
RESEARCH FOCUSED ON RNA FOR HIS
ENTIRE CAREER AND MOST RECENTLY
ON THE POSITIVE SENSE RNA
VIRUSES AND PATHOGENESIS. DR.
GEHRKE WAS ELECTED FELLOW OF THE
AMERICAN INSTITUTE OF MEDICAL
AND BIOLOGICAL ENGINEERING IN
2021. AND AS A FELLOW OF THE
AMERICAN ACADEMY OF MICROBIOLOGY
IN 2022. DR. GEHRKE WILL BE
SPEAKING ON DEFINING VIRAL
PATHOGENESIS MECHANISMS USING
HUMAN CEREBRAL ORGANOIDS. THANKS
SO MUCH. OVER TO YOU.
>> THANK YOU FOR THAT
INVITATION. ARE MY SLIDES
VISIBLE, CAN YOU HEAR ME?
>> ALL LOOKS GOOD. THANK YOU.
>> FABULOUS. SO TODAY I WANT TO
TELL YOU ABOUT SOME OF THE WORK
WE ARE DOING ON VIRAL
PATHOGENESIS USING HUMAN
CEREBRAL ORGANOID MODELS. I WILL
START BY ADVANCING MY NEXT
SLIDE. I WILL START BY SAYING
THAT MY LAB IS REALLY A CONSUMER
OF THIS TECHNOLOGY AND I WANT TO
POINT OUT THE PEOPLE WHO HAVE
REALLY BEEN PIONEERS IN
DEVELOPING ORGANOID TECHNOLOGY.
WE ARE USING TECHNIQUES THAT
WERE PIONEERED BY MADELINE
LANCASTER. WE ALSO HEARD FROM
HANS CLEVER YESTERDAY ANOTHER
PERSON WHO IS DONE A GREAT DEAL
TO DEVELOP THIS TECHNOLOGY BUT
TODAY I'M GOING TO TELL YOU
ABOUT WORK WE HAVE DONE AND
ABOUT THE LAST FIVE YEARS. SO IF
ANY OF YOU ARE THINKING ABOUT
USING ORGANOID SYSTEMS
PARTICULARLY CEREBRAL ORGANOIDS
I WANT TO TELL YOU ABOUT WHAT WE
HAVE LEARNED IN USING THESE
SYSTEMS, COMING FROM NEVER
HAVING GROWN ONE UNTIL FIVE
YEARS AGO. I THINK MANY PERHAPS
ALL OF US ARE IN THIS WORKSHOP
ARE INTERESTED IN STUDYING
HALLWAYOLOGY. TO DO THAT IT IS
NOT APPROPRIATE TO DO
EXPERIMENTATION ON HUMANS. SO WE
OFTEN TURN TO USING ANIMAL
MODELS. ANIMAL MODELS ARE
EXTREMELY IMPORTANT, HAVE BEEN
AND WILL CONTINUE TO BE
IMPORTANT BUT THERE ARE EXAMPLES
OF HOW ANIMAL MODELS DON'T
COMPLETELY RECAPITULATE THE
HUMAN BIOLOGY. AS A VIROLOGIST
THERE ARE MANY VIRUSES FOR WHOM
RODENTS ARE NOT NATURAL HOSTS.
SO IN ORDER TO BE ABLE TO STUDY
THEM IT IS NECESSARY TO USE MICE
THAT HAVE THEIR SYSTEM FOR
EXAMPLE KNOCKED OUT SO IT
DOESN'T RECAPITULATE COMPLETELY
SO WHAT WE WANT TO DO WAS TO
FIND A SYSTEM THAT WAS A BRIDGE.
A BRIDGE BETWEEN HUMAN BIOLOGY
AND THE ANIMAL MODELS THAT'S
WHEN WE STARTED USING ORGANOID
SYSTEMS WE STUDY NEUROTROPIC
VIRUSES SO WE WERE INTERESTED IN
USING CEREBRAL ORGANOID
TECHNOLOGY SO THE IMAGE IN THE
CENTER HERE IS ONE OF A ORGANOID
DERIVED FROM HUMAN PLURIPOTENT
STEM CELLS. SO THIS IS A SLIDE
THAT'S ALREADY BEEN SHOWN IN A
NUMBER OF CASES THROUGHOUT THIS
WORKSHOP. WE CREATE THE
ORGANOIDS IN PRETTY MUCH THE
SAME WAY. FROM STARTING WITH
STEM CELLS TO THE POINT WHERE WE
DO ALL INFECTIONS TAKES 21 DAYS
AND THE ORGANOIDS AT THAT POINT
ARE ABOUT 2-MILLIMETERS IN
DIAMETER. IF YOU SECTION THROUGH
CEREBRAL ORGANOID IT LOOKS LIKE
IMAGES THAT ARE BELOW HERE AT
DAY 17, DAY 21 AND DAY 28. YOU
CAN SEE THEY GROW, THESE ARE
SAME SCALE. ID WILL SHOW IMAGES
THAT HAVE THIS TYPE OF STAINING
AND SO THROUGHOUT THE ORGANOIDS
YOU CAN SEE ROUND STRUCTURE WITH
OPENING IN THE CENTER THESE ARE
VENTRICLES RECAPITULATE LAYERING
FOUND IN CORTEX. THEY ARE
SURROUNDED BY WITH SOX 2
TRANSCRIPTION FACTOR. THEN YOU
CAN ALSO SEE THE GREEN STAINING
MAP 2 NEURONS BLUE STAINING IS
DAPE. IF YOU WANT TO THINK ABOUT
HOW THESE ORGANOIDS CORRELATE
WITH DEVELOPMENT OF HUMAN BRAIN,
LANCASTER PUT TOGETHER THIS
SLIDE AND ON THE TOP HERE SHOWS
THE TIMETABLE FOR DEVELOPMENT OF
HUMAN BRAIN THROUGH FIRST AND
SECOND AND THIRD TRIMESTERS AND
THEN ON THE BOTTOM HERE IS
DEVELOPMENT OF CEREBRAL
ORGANOIDS FROM 20 TO 40 TO 60
DAYS. IN THE CENTER THERE IS A
COLOR GRAPH THAT SHOWS WHICH
POINTS THERE'S BEST CORRELATION
BETWEEN BIOLOGY OF THE ORGANOID
AND THE HUMAN BRAIN. THAT IS IN
THIS PURPLISH REGION RIGHT HERE.
AND IN FACT WE DO OUR INFECTIONS
TO STUDY VIRAL PATHOGENESIS FROM
21 DAYS TO 41 DAYS. SO THIS IS
THE BIOLOGICAL PROBLEM WE ARE
INTERESTED IN. WE ARE INTERESTED
IN UNDERSTANDING HOW VIRUSES
THAT ARE CLOSELY RELATED CAN
CAUSE SUCH DIFFERENT PATHOLOGIES
AND SOME MAKE PEOPLE SICK,
OTHERS DON'T MAKE PEOPLE SICK.
AND AN EXAMPLE OF THAT SHOWN IN
THESE IMAGES IS HERE. ON THE
LEFT IS THE MOCK INFECTED
ORGANOID, THE PINK COLOR IS
VENTRICLES THAT CONTAIN
PROGENITOR CELLS ORAL PROGENITOR
CELLS, GREEN IS NEURONS AND BLUE
STAIN IS DAPE NUCLEI. HERE IS
UNINFECTED. THIS IS ONE STRAIN
OF ZIKA THAT IS I WOULD SAY A
MILD PATHOGENESIS. THE ORGANOIDS
RETAIN THEIR OVERALL STRUCTURE
AND ONE CAN ALSO SEE THE
VENTRICLES, THE WHITE DOTS HERE
ARE VIRUS, OR VIRAL RNA, THEY
ARE STAINED WITH AN ANTI-DOUBLE
STRANDED RNA ANTIBODY. THEN WE
GO TO THE SECOND STRAIN WHICH IS
MORE PATHOGENIC. WHEN WE USE
THIS VIRUS EVEN THOUGH IT IS 98%
IDENTICAL TO THIS ONE HERE AT
THE MOS LEVEL WE LOST A LOT OF
VENTRICLE ARCHITECTURE, THE
NEURONS HAVE BEEN DISRUPTED WE
SEE CELL BODIES BUT WE DON'T SEE
AXONS AND THEN WE GO TO THE
THIRD VIRAL STRAIN ZIKA STRAIN
WHICH IS VERY PATHOGENIC AND WE
LOST COMPLETELY THE ORGANIZATION
OF THE VENTRICLES THERE IS
LITTLE MAP Q STAINING ON -- THAT
CAN BE SEEN. SO ONE BIG
ADVANTAGE OF THAT WE FIND USING
THIS ORGANOID SYSTEM IS THAT WE
CANNOT ONLY DO THE MOLECULAR
BIOLOGY IN THE BIOCHEMISTRY ON
THEM, BUT SEEING WHAT HAPPENS TO
THE TISSUE, THIS IS A COMPLEX
THREE DIMENSIONAL TISSUE, AND
SEE WHAT HAPPENS TO THE
ORGANIZATION OF THE INDIVIDUAL
CELLS IS VERY IMPORTANT. SO NOW
I WANT TO GIVE A FEW TIPS WE
HAVE LEARNED SOMETIMES THE HARD
WAY IN DOING EXPERIMENTATION
USING CEREBRAL ORGANOIDS, AGAIN
FOR ANYONE WHO IS IN THE
AUDIENCE WHO MIGHT BE THINKING
ABOUT ADOPTING THIS TECHNOLOGY I
WOULD ENCOURAGE YOU TO DO SO.
BUT I WANT TO TELL YOU ABOUT
SOME OF THE THINGS WE HAVE
LEARNED ALONG THE WAY. SO MY
FIRST TIP IN USING CERTAIN
ORGANOID OR ORGANOID MODELS IN
GENERAL IS TO IDENTIFY STEM CELL
COLLABORATOR. IN OUR CASE, WE --
OUR COLLABORATOR IS RUDOLPH, A
PIONEER IN THE STEM CELL FIELD,
HE IS JUST ACROSS THE STREET
FROM US AT THE WHITE HEAD
INSTITUTE AND COLLABORATING WITH
HIS LAB ON VIRUS INFECTION AND
STEM CELLS HAS BEEN PRODUCTIVE
BUT HAS BEEN ABSOLUTELY CRITICAL
TO HAVE THEIR EXPERTISE AS STEM
CELL BIOLOGISTS. THE REASON IS
THAT STEM CELLS AND STEM CELL
BIOLOGY IS BIG FIELD. IT IS
CHALLENGING FOR SOMEONE WHO IS
NOT A STEM CELL BIOLOGIST LIKE
ME TO ACQUIRE INFORMATION SO
QUICKLY THAT THIS CAN BE DONE
COMPLETELY INDEPENDENTLY. IT
DEFINITELY SHOWS IT IS A
CHALLENGE AND SOMETHING TO BE
VOIDED AS WORKING IN IN A VACUUM
AND DEVELOPING THESE THINGS
COMPLETELY ON YOUR OWN. PEOPLE
SKILLED CAN DO IT BUT YOU CAN DO
IT FASTER WITH GREATER
UNDERSTANDING IF YOU HAVE A
COLLEAGUE AND FINALLY STEM CELL
LABS HAVE SOME PIECES OF
EQUIPMENTNA ARE SPECIALIZED THAT
ARE VERY -- THAT ARE SPECIALIZED
VERY IMPORTANT. IN OUR CASE IT
IS NOT THIS ONE, THIS IS JUST A
PICTURE BUT STEM CELL LABS HAVE
LOW OXYGEN INCUBATORS THAT MY
LAB DOESN'T HAVE HAVE. AND THEYE
USED FOR KEEPING THE THEY ARE
USE TO KEEP THE STEM CELLS FROM
DIFFERENTIATING. THE SECOND TIP
IS TO USE HIGH QUALITIES
PLURIPOTENT STEM CELLS. THIS IS
REALLY THE FOUNDATION FOR
EVERYTHING DONE HER HERE. THIS E
OF GARBAGE IN AND OUT COULDN'T
BE MORE TRUE WHEN DOING THESE
PLURIPOTENT STEM CELL EXPERI
EXPERIMENTS. THIS IS ALSO
CRITICAL FOR PRECISION, ACCURACY
REPRODUCIBILITY OF YOUR
EXPERIMENTS TO MAKE SURE YOU DO
EVERYTHING YOU CAN TO HAVE GOOD
STARTING AND WELL CHARACTERIZED
STARTING MATERIAL. AS WE HAVE
DONE THESE EXPERIMENTS YOU
ACQUIRE EXPERTISE AND NOTICE
THINGS THAT ARE WARNING SIGNS.
SO ON THE LEFT HERE IS AN
ORGANOID WE WOULD LIKE TO SEE ON
THE RIGHT THOUGH MAY NOT BE
OBVIOUS FIRST GLANCE WHY THIS IS
NOT ONE THAT WOULD WANT, WE
WOULD WANT. SO ON THE LEFT YOU
CAN SEE THE EMBRYO BODY HERE AND
THE CLEAR AREA AROUND THE
DEVELOPING EMBRYO BODY IS NEUTRA
GEL. T NOTICE ON THE RIGHT THAT
THERE ARE PROJECTIONS THAT ARE
GOING OUT INTO THE NATRIGEL.
THESE ARE HARRY ORGANOIDS. WE
DON'T WANT THESE AND WE LEARNED
THE HARD WAY THAT IF THE
ORGANOIDS HAVE THESE PROJECTIONS
IN THE NATRIGEL THEY HAVE
DIFFERENCE ADJUDICATED
PREMATURELY AND NOT USEFUL. SO
IF WE EVER SEE THIS KIND OF
ORGANOID IN OUR PREPS WE JUST
START OVER AGAIN. PREMATURE
DIFFERENTIATION OF THE
PLURIPOTENT STEM CELLS IS
SOMETHING ONE HAS TO KEEP IN
MIND, THIS IS WHERE
ORGANOIDS. AND YOU CAN
DIFFERENTIATE THE STEM CELLS
INTO THESE PROGENITORS AND THEN
FREEZE THEM AND STORE THEM FOR
UPCOMING EXPERIMENTS. IF YOU
HAVE A GREAT PREP OF THE NEURAL
PROGENITORS YOU CAN FOLLOW THEM
AND YOU CAN GROW THEM UP,
PASSAGE THEM FOR SEVEN OR EIGHT
PASSAGES. IF IT IS NOT A GREAT
PREP THEY WILL START TO
DIFFERENTIATE INTO OTHER
STRUCTURE WITHIN THE FIRST
PASSAGE. AND YOU LOSE THE
ABILITY TO GET ENOUGH CELLS TO
DO YOUR EXPERIMENT. HAVING GREAT
STARTING MATERIAL IS SO
IMPORTANT. ANTIBIOTIC. THERE'S
NOTHING THAT RUINS YOUR DAY MORE
THAN COMING IN IN THE MORNING
AND FINDING OUT THAT YOUR
CULTURES ARE CONTAMINATED. IN
THE CASE OF THE ORGANOID FIELD,
MOST OF THE PAPERS AND INDEED
THE MANUFACTURERS LIKE STEM CELL
TECHNOLOGIES DO NOT USE
ANTIBIOTICS. IN OUR WORK WE
GENERALLY -- LEAVE ANTIBIOTICS
OUT AS WELL. BUT THERE ARE CASES
WHEN YOU ARE TRAINING A NEW
PERSON, SOMEBODY NEW IS COMING
IN, THERE ARE A LOT OF
OPPORTUNITIES FOR CONTAMINATING
CULTURES PARTICULARLY IN
EMBEDDING STEP. SO WE HAVE USED
THEM AND WE HAVEN'T -- WE
HAVEN'T FOUND NEGATIVE EFFECTS
WITH CALLED STEM CELL
TECHNOLOGIES AND THEY SAY THAT
THEY DON'T RECOMMEND THE OUTWARD
USE OF ANTIBIOTICS HOWEVER THEY
USED THEM TRIED THEM AND HAVEN'T
SEEN ANY DELETERIOUS EFFECT. SO
THE BOTTOM LINE IS THAT I THINK
IT IS POSSIBLE TO USE THEM, WE
TRY TO AVOID IT WHENEVER WE CAN
BUT WE HAVE NOT SEEN ANY
DELETERIOUS EFFECTS OF
ANTIBIOTICS. THE NEXT ONE HERE
IS BE GENTLE AND THIS IS -- THIS
COULD BE ONE'S LIFE MANTRA. BUT
IN TERMS OF DOING ORGANOIDS, IT
IS IMPORTANT AND WHEN
TRANSFERRING THESE FROM ONE
PLATE TO ANOTHER, YOU DON'T WANT
TO SUCK THEM UP INTO A NARROW
BOW BY PET TIP BECAUSE THEY ARE
FRAGILE, YOU WILL DAMAGE THEM SO
WE TAKE ONE PIPETTE TIP, CUT
THEM OFF WITH A CLEAN RAZOR AND
USE THOSE. THE GREEDY PART IS
WHEN YOU ARE TRANSFERRING INTO
LARGER DISHES FOR EXAMPLE YOU
WANT TO CHANGE MEDIA, WE WOULD
ADVISE TO RESIST THE TRYING TO
GET EVERY LAST MICROLITER OF
MEDIA OUT OF THE DISH BECAUSE
YOU ARE LIABLE TO DAMAGE THE
ORGANOIDS OR ALLOW THEM TO DRY
OUT BOTH OF WHICH WILL RUIN YOUR
EXPERIMENT. BOTTOM LINE HERE IS
THAT ONE HAS TO BE CAREFUL AND
PATIENT IN WORKING WITH THESE
ORGANOIDS. ANOTHER PROBLEM THAT
WE HAVE HAD FROM TIME TO TIME IS
THAT THESE ORGANOIDS WILL FUSE
IF THEY GET IN CONTACT WITH ONE
ANOTHER. IF THE ORGANOIDS GET
TOO CLOSE THEY WILL FUSE INTO
ANOTHER ORGANOID AND
UNFORTUNATELY WHEN THAT HAPPENS,
THEY USUALLY DON'T LOOK LIKE
THIS NICE ROUND ORGANOID, THEY
ARE VERY IRREGULAR IN SHAPE. WE
TRY TO AVOID THAT. SO WHAT WE DO
IS PUT THEM INTO 24 WELL PLATE
WELLS AND CULTURE THE ORGANOIDS
INDIVIDUALLY SO THEY DON'T HAVE
THE OPPORTUNITY TO DIFFUSE WITH
ONE ANOTHER. SO THIS ONE IS
OBVIOUS BUT ONE NEEDS TO REDUCE
EXPERIMENTAL DELIVERY AND WE DO
THAT WITH DOING BATCH
EXPERIMENTS SO WE DO A LOT OF
VIRUS INFECTIONS SO WE TAKE A
GROUP OF ABOUT 15 TO 20
ORGANOIDS, PUT THEM INTO A SIX
WELL PLATE AND DO THE INFECTION
OF ALL THE ORGANOIDS AT ONCE.
THAT REALLY REDUCE IT IS
VARIABILITY IN EXPERIMENTS
OPPOSED TO TRYING TO TAKE THREE
OR FOUR ORGANOIDS AND INFECT
THEM SEPARATELY. THAT WOULD
CHARACTERIZE ANY EXPERIMENTAL
PROTOCOL THAT YOU ARE USING. IF
YOU ARE DOING SECTIONS WE DO
SECTIONING TO SEE WHAT THE
ORIGINATION OF THE ORGANOIDS
LOOK LIKE. A HELPFUL SMALL TIP
IS USE GLUE OCT. THIS COMES IN A
NUMBER OF COLORS BUT THE
ORGANOIDS ARE VERY SMALL AND
WHITE AND YOU GET THEM INTO A
BLOCK IT IS VERY HARD TO SEE
THEM. SO THAT YOU KNOW WHERE TO
START CUTTING AND SO IF YOU USE
BLUE OCT THAT REALLY HELPS WITH
VISUALIZING THEM. IF YOU WANT
TO DO SINGLE CELL RNA SEQ WE
HAVE DONE A LOT OF SINGLE CELL
RNA SEQ ON THESE ORGANOIDS
DISSOCIATING THE ORGANOIDS AND
RETAINING VIABILITY IS CRITICAL
AS IS TRUE FOR ANY SINGLE CELL
SEQ BUT WE FOUND THIS PARTICULAR
KIT, I DON'T WORK FOR THEM NOR
DO THEY GIVE ME ANYTHING BUT
THIS EM BRIEFED YOU BODY
DISSOCIATION KIT GAVE US THE
GREATEST VIABILITY OF CELLS WE
CAN CARRY FORWARD THEN INTO
SINGLE CELL SEQUENCING. SO TO
SUMMARIZE THESE POINTS WE LIKE
TO SEE AT THE END THE ORGANOIDS
ARE ROUND, BUT THEY CAN VARY BY
ABOUT TWOFOLD IN DIAMETER NO
MATTER HOW WE DO THE PREPS, THEY
HAVE APPROPRIATE -- WILL VARY
FROM SIZE BY FACTOR OF TWO. WE
GET USABLE ORGANOIDS 80 TO 90%
OF THE TIME, WHICH IS PRETTY
GOOD FOR THE TECHNOLOGY AND WE
DO BATCHES OF 300. THE 80 TO 90%
TIME WOULD NOT INCLUDE HAVING A
CONTAMINATION PROBLEM IN THE
LAB, THAT CAN BE FIXED AND IS
NOT INCLUDED IN THESE NUMBERS
BUT FOR JUST THE TECHNIQUE WE
CAN GET GOOD RESULTS WITH THE
ORGANOID PREPS QUITE OFTEN. SO
ONE OF THE VARIABLES THAT CAN
AFFECT OUTCOMES SOME OF THESE WE
SUSPECT WE DON'T REALLY HAVE
HARD DATA TO KNOW WHAT THE
PROBLEMS ARE BUT THE QUALITY OF
THE PLURIPOTENT STEM CELLS IS
ABSOLUTELY CRITICAL FOR GETTING
GOOD RESULTS. YOU WANT TO AVOID
CONTAMINATION AS I TOLD YOU
BELOW HERE. INFORMING THESE
ORGANOIDS THEY HAVE TO HAVE A
SOLID MATRIX AND THAT IS DONE BY
USING NATRIGEL AND THE LOTS OF
NATRI-- NATRIJELL VARY. THIS WAS
DISCUSSED BY LINDA GRIFFITHS
HANDS IS SOMETHING TO KEEP IN
MIND. MOST MEDIA WE USE IS MADE
COMMERCIALLY BY STEM CELL
TECHNOLOGIES, BUT THERE ARE
OTHER PARTS OF IT ONE HAS TO
MAKE IN THE LAB AND THERE CAN BE
VARIABILITY IN THOSE PREPS. THE
OTHER THING TO MENTION, IT IS IN
DEVELOPING EXPERTISE AND DOING
THIS THE LOWER THE AMOUNT OF
HANDLING YOU CAN DO WITH THESE
ORGANOIDS THE BETTER OFF. THEY
ARE FRIABLE, FRAGILE AND IF YOU
CAN MINIMIZE NUMBER OF STEPS YOU
WILL GET BETTER RESULTS. SO
COMING BACK THEN TO DRUGS AND
THESE ORGANOIDS, I HAVE
MENTIONED IN PREVIOUS SLIDE
INTEREST AND UNDERSTANDING HOW
THIS IS THE EXAMPLE WE SEE IN
THE SHY DA VIRUS STRAINS AND
PATHOGENIC VIRUSES THERE IS LOSS
OF THIS ORGANIZATION AND LOSS OF
NEURAL PROGENITOR CELLS. SO WE
HAVE BEEN LOOKING AT A FEW
COMPOUNDS THAT MIGHT AFFECT US
AND AFFECT THIS TECHNOLOGY AND
HELP US UNDERSTAND WHAT IS GOING
ON. SO THIS IS A SET OF
EXPERIMENTS WE DID, I'M SHOWING
YOU ADDITION O VITAMIN E THE
WHOLE ORGANOIDS ARE SHOWN AT THE
TOP AND THE BOXES ARE THE INSETS
SHOWING PICTURES, WITHOUT
TREATMENT. IN THE MOCK WE SEE
THESE NICE VENTRICLES AND
NEURONS, IF WE ADD VITAMIN E NOT
MUCH CHANGE. THE VENTRICLES
REMAIN INTACT. IF WE ADD THE
LEAST PATHOGENIC STRAIN NOW YOU
CAN SEE THE WHITE DOTS WHICH ARE
VIRUSES THAT ARE PRESENT IN
INFECTION. WE STILL SEE WELL
ORGANIZED VENTRICLES IN THE
VEHICLE AND THEN IF WE ADD
VITAMIN E. THERE IS SOME
IMPROVEMENT IN THE STRUCTURE BUT
WE DON'T SEE DISRUPTION TOO MUCH
OF THE VENTRICLES WITH THIS
VIRUS. IF WE GO TO MOST
PATHOGENIC VIRUS WE SEE HERE,
THIS IS THE WHOLE ORGANOID, WE
LOSE ALL THE ORGANIZED
VENTRICLES. WE SEE SOME SOX 2
STAININGS, VERY LITTLE MAP 2
STAINING BUT IF WE ADD VITAMIN E
YOU CAN SEE IN THE WHOLE PICTURE
AT THE TOP HERE NOW WE SEE
VENTRICLES WHERE WE DIDN'T SEE
THEM HERE. AND WE CAN RESCUE
THIS PHENOTYPE BY ADDING SOME
VITAMIN E. SO WE THINK THIS IS A
GOOD MODEL SYSTEM UNDERSTANDING
HOW SOME OF THESE DRUGS WORK. TO
SUMMARIZE THEM WE PROPOSE
CEREBRAL ORGANOID MODELS A GOOD
BRIDGE BETWEEN HUMAN BIOLOGY AND
ANIMAL CELL MODELS AND PERMIT
EXPERIMENTATION IN A COMPLEX 3
DIMENSIONAL CELLULAR
ENVIRONMENT. IF ONE IS
INTERESTED IN USING THIS
TECHNOLOGY, MASTERING THE
FUNDAMENTALS, I WOULD SAY IS
VERY, VERY IMPORTANT SO YOU ARE
ALWAYS STARTING PARTICULARLY
WITH GOOD STARTING MATERIALS,
IPSCs OR HUMAN PLURIPOTENT
STEM CELLS, IN TERMS OF GETTING
GOOD REPRODUCIBLE RESULTS. THIS
IS NOT A SLASH AND BURN
TECHNOLOGY. I WOULD SAY IT TAKES
TIME AND ACTION CARE IN HANDLING
THE ORGANOIDS IN ORDER TO BE
ABLE TO GET GOOD REPRODUCIBLE
DATA. FINALLY I CLOSE BY SAYING
THAT THESE ORGANOIDS WE BELIEVE
ARE REALLY GREAT MODELS FOR
LOOKING AT THE PATHOGENESIS OF
NEUROTROPIC RNA VIRUSES. SO I
WANT TO ACKNOWLEDGE THE PEOPLE
IN THE LAB WHO DID THE WORK ON
THE LEFT HERE. AND ON THE RIGHT
I WANT TO ACKNOWLEDGE OUR
COLLABORATOR IN THIS WORK AND
THEN OUR FUNDING IS HERE. THANK
YOU VERY MUCH FOR YOUR
ATTENTION. AT APPROPRIATE TIME I
WILL BE HAPPY TO ANSWER YES,
SIYEQUESTIONS.
>> EXCELLENT. THANK YOU SO MUCH,
DR. GEHRKE. EXCELLENT
PRESENTATION. WE LOOK FORWARD TO
THE DISCUSSION OF QUESTIONS
COMING UP THIS AFTERNOON. NOW I
WOULD LIKE TO TURN TO OUR THIRD
SPEAKER, OF THE SESSION,
INTRODUCE DR. SASHI RAMANI,
ASSISTANT PROFESSOR DEPARTMENT
OF MOLECULAR VIROLOGY AND
MICROBIOLOGY AT BAYLOR COLLEGE
OF MEDICINE. MATERNAL AND CHILD
HEALTH WITH FOCUS ON GASTRO
INTESTINAL INFECTION AND
VACCINES. SHE HAS EXPERIENCE
WORKING ON EPIDEMIOLOGY AND
PATHOGENESIS OF ROW D TA VIRUSES
AND NORA VIRUSES WITH THE GOAL
OF FACTORS THAT CONTRIBUTE TO
DISEASE SUSCEPTIBILITY AND
IDENTIFY MECHANISMS TO IMPROVE
IMMUNE RESPONSE AND TO
INFECTIOUS AGENTS AND VACCINE
DEVELOPMENT. WORK USING HUMAN
INTESTINAL ORGANOID CULTURES
FROM DIVERSE DONORS PROVIDES
PHYSIOLOGICALLY RELEVANT AND
GENETICALLY REPRESENTATIVE
SYSTEM TO UNDERSTAND HOST
PATHOGEN INTERACTIONS AT
MOLECULAR LEVEL AND ENABLES
PRE-CLINICAL TESTING OF
INTERVENTIONS THROUGH
COLLABORATIVE -- COLLABORATION
WITH INTERDISCIPLINARY GROUP OF
RESEARCHERS AND THROUGH
POPULATION STUDIES TO EVALUATE
FINDINGS IN HUMAN SUBJECTS. DR.
RAMANI'S WORK TAKES A BENCH TO
BEDSIDE APPROACH TO
UNDERSTANDING ENTERIC INFECTIOUS
DISEASE AND TODAY DR. RAMANI
WILL BE SPEAKING ON THE TOPIC OF
HUMAN INTESTINAL ORGANOIDS AS A
PLATFORM FOR TESTING ANTIVIRALS
TO HUMAN NOR ROW VIRUS.
>> THANK YOU, I WANT TO THANK
THE ORGANIZERS FOR THE
INVITATION TO BE PARTS OF THIS
MEETING. I'M VERY EXCITED TO
SHARE WITH Y'ALL SOME OF OUR
WORK ON USING HUMAN INTESTINAL
ORGANOIDS AS A PLATFORM FOR
ANTIVIRAL TESTING FOR HUMAN
NOROVIRUS. AS A BRIEF INVESTOR
EWE OF WHAT IS TO COME IN THESE
20 MINUTES I WILL DO A QUICK
INTRODUCTION TO HUMAN NOROVIRUS
AND TALK ABOUT THE HISTORICAL
PERSPECTIVE ON CHALLENGES THAT
WE HAD WITH GROWING THIS VIRUS
IN THE LAB. AND HOW ANDROIDS ARE
USEFUL TO STUDY VIRAL
REPLICATION. IN THE CONTEXT OF
ANTIVIRAL, I WANT TO TELL YOU
PIPELINE WE HAVE IN THE LAB AND
WE WERE CHARGED WITH TALKING
ABOUT SOME OF THE CHALLENGES WE
FACE. SO I WILL TALK ABOUT SOME
CHALLENGES IN CYTOTOXICITY
ASSESSMENT THAT WE ENCOUNTER
DOING ANTIVIRUS STUDIES AND GIVE
UPDATES ON PROGRESS IN WHERE WE
ARE WITH TESTING DRUGS FOR
NOROVIRUS. AND I LIKE WHERE THE
ORGANOID FIELD IS MOVING, IN
TERMS OF DEVELOPING COMPLEX
CULTURES FOR INFECTIOUS DISEASE
WITH THAT AS A AN INTRODUCTION
TO HUMAN NOROVIRUS. IT IS A
LEADING CAUSE OF THE GASTRO
ENTERITIS ACROSS ALL AGE GROUPS,
DIARRHEA, VOMITING, SOMETHING
YOU DO NOT WANT TO HAVE. IT HAS
A FAIRLY BROAD SPECTRUM OF
ILLNESS, IN THAT IF YOU TAKE
IMMUNOCOMPETENT HOST ACROSS ALL
AGES YOU SEE ACUTE -- PEOPLE GET
SICK AND SYMPTOMS RESOLVE IN A
COUPLE OF DAYS BUT WHEN YOU LOOK
AT IMMUNOCOMPROMISED PATIENTS
WHETHER CANCER PATIENTS OR
TRANSPLANT PATIENTS, INFECTIONS
ARE CHRONIC IN THAT NOROVIRUS
DIARRHEA CAN LAST YEARS AND YOU
CAN IMAGINE HOW THIS WOULD
AFFECT QUALITY OF LIFE OF THE
PERSON WHO ALREADY HAS A
CONDITION LIKE CANCER OR HAS HAD
M. IF YOU LOOK AT MORTALITY
BECAUSE OF THIS VIRUS IN
EXTREMES OF AGE GROUPS, SO YOUNG
CHILDREN AND OLDER ADULTS AND
THERE ARE ABOUT 200,000 DEATHS
ANNUALLY. SO ALL THIS TELLS YOU
HUMAN NOROVIRUS IS A PROBLEM A
BIG HEALTHCARE PROBLEM. THIS IS
REALLY EXPENSIVE VIRUS. IN THAT
IT IS ABOUT $60 MILLION PLUS $60
BILLION SPENT IN BECAUSE OF
NOROVIRUS AND MAJORITY OF THESE
COSTS COME FROM COSTS LIKE
PARENTS HAVING TO TAKE TIME OFF
WORK BECAUSE DAY CARE IS CLOSED
BECAUSE OF OUTBREAK OR HAVING TO
CLOSE RESTAURANT BECAUSE THERE
IS NOROVIRUS INFECTIONS AND YOU
HAVE TO SHUT DOWN THE
RESTAURANT, ET CETERA. AT THIS
POINT IN TIME THERE ARE NO
LICENSED VACCINES OR
THERAPEUTICS AND WE HAVE COME TO
THAT IN A MINUTE. AND I DID WANT
TO SAY THAT IN THE MEDIA
NOROVIRUS IS ALWAYS KNOWN AS THE
CRUISE SHIP VIRUS BUT IT IS MORE
THAN THAT. CRUISE SHIPS ACCOUNT
FOR A VERY SMALL PERCENTAGE OF
NOROVIRUSES CASES AND IN FACT
MAJORITY OF CASES COME FROM
HEALTHCARE FACILITIES,
RESTAURANTS, DAY CARES, ET
CETERA. AND THE VEHICLE FOR
TRANSMISSION IS CONTAMINATED
FOOD AND WATER. SO SORRY TO
SPOIL YOUR LUNCH BUT LOTS OF
PEOPLE SAY IT IS FOODS, OYSTERS,
DELL LIVE MEAT CONDITION
CONTAMINATED WITH HUMAN
NOROVIRUS BUT ALSO IN
RECREATIONAL WATERS, YOU HEAR
STORIES OF THE ENTIRE FOOTBALL
TEAM COMING DOWN WITH NOROVIRUS.
AND PART OF THE REASON WHY WE
HAVE NOT HAD AN ANTIVIRAL OR
VACCINE FOR THIS VIRUS WAS THE
CHALLENGE IN STUDYING THIS VIRUS
IN THE LAB. SO THIS VIRUS, THE
FIRST KNOWN VIRAL CAUSE OF
GASTROENTERRYETIS WAS VISUALIZED
BY (INAUDIBLE) IN 1972 USING
CULTURES FROM ELEMENTARY SCHOOL
OUTBREAK IN NOR WALK OHIO IN
1968. WE STUDIED THIS VOICE FOR
DECADES SUMMARIZED BY THIS PAPER
THAT CAME OUT IN 2004 WHICH
LOOKED AT EFFORTS TO CULTIVATE
THIS VIRUS IN OVER 20 DIFFERENT
CELL LINES. THIS IS THE SUMMARY
-- THAT ALL CHANGED IN FIVE TO
SEVEN YEARS AGO WHEN DIFFERENT
MODELS OF THIS VIRUS BEGAN TO BE
DESCRIBED. WE DESCRIBE THE HUMAN
INTERESTAL ANDROID MODEL IN 2016
AND MORE RECENTLY ZEBRAFISH
MODEL. I WILL TALK ABOUT USING
ANDROIDS TO STUDY HUMAN
NOROVIRUS AS INTESTINAL FOR THIS
PATH GENERAL. TALKING BRIEF --
PATHOGEN. THIS AUDIENCE NEEDS NO
INTRODUCTION BUT IF YOU THINK
ABOUT ORGANOID CULTURE MS. THE
CONTEXT OF THE GUT YOU HAVE
TISSUE DERIVED CULTURES AND
CULTURES THAT COME FROM
PLURIPOTENT STEM CELLS.
PLURIPOTENT STEM CELLS HAVE A
STROMAL MESENCHYME, OUR EARLY
WORK SHOWS IN FACT THESE
CULTURES ARE NOT SUSCEPTIBLE TO
HUMAN NOROVIRUS INFECTION. SO IN
OUR LAB WHAT WE WORK WITH ARE
BASICALLY TISSUE DERIVED
CULTURES FROM INFANTS, ADULTS,
GOOD SPECIMENS WE ISOLATED
THESE, 3D STRUCTURE AND WE
PLAYED THEM AS MONOLAYERS ON 96
WELL PLATE. AS WE HEARD
YESTERDAY, IDENTIFY THIS
TECHNOLOGY, THESE REMARKABLE
CULTURES NOT TRANSFORMED TO
PASSAGE INDEFINITELY THEY HAVE
THE DIFFERENT EPITHELIAL CELL
TYPES. AND THEIR PHYSIOLOGICALLY
RESPONSIVE TO INFECTI INFECTIONN
MAKE THEM DIFFERENT INFECTIONS
OF THE GUT AND THEY DEEM
COMPLEXITY AND ORGANIZATION OF
THE EPITHELIUM THEY COME FROM.
SO WE SHOWED IN 2016 HIE SHOW
SUPPORT OF MULTIPLE VIRUSES. WE
TOOK TWO EXAMPLES A DOMINANT
STRAIN OF NOROVIRUS AND IT HAS
DIFFERENT PANDEMIC VARIANTS. IF
YOU LOOK AT THE RTPCR YOU SEE IN
THE LIGHT BLUE IS BINDING AND
HOW MUCH INFECTION VIRUSES
PRODUCE AT 72 HOURS AND YOU CAN
SEE HERE THAT THERE IS INCREASE
IN REPLICATION FROM ONE FOR ALL
THESE DIFFERENT VIRUSES WE LOOK
BY IMMUNOFLUORESCENCE STAINING
FOR THE CAPSID AND YOU CAN SEE
20% INFECTED CELLS, YOU CAN SEE
THAT ALSO IMMUNOFLUORESCENCE AND
IN SOME CASES WE WILL SEE
CYTOKINE EFFECTS. SO WE
ESSENTIALLY SHOWED REPLICATION
FOR A FEW VARIANTS, THE
PROTOTYPE VIRUS THAT CAME FROM
GP 1 STRAIN AND TWO OTHERS. AND
SINCE THEN WE HAVE GONE ON TO
EXPAND PROFILE OF NOROVIRUS. AND
WE HAVE DONE THIS BY WORKING
WITH MEDIA, TRAINING MEDIA
COMPOSITIONS AND WE WORK WITH
MAKING GENETICALLY QUANTIFIED
ORGANOIDS. COMPLEX EPIDEMIOLOGY,
THERE ARE MANY DIFFERENT GENOME
GROUPS, ONE GENOME GROUP WITHIN
GENOTYPES SOME GENOTYPES HAVE
VARIANTS, AS I MENTION THE
PREVIOUS SLIDE YOU HAVE PANDEMIC
VARIANTS THAT EVOLVE EVERY FEW
YEARS. SO NOW WE HAVE A FAIRLY
LARGE BATCH OF STRAINS AND WE
HAVE BEEN ABLE TO SHOW THAT
NOROVIRUS CAN GROW IN ALL
SEGMENTS OF THE INTESTINE,
DUODENUM, ILEUM BUT DOESN'T
REALLY GROW (INAUDIBLE). THE
OTHER REASON THESE CULTURES ARE
MULTIPLE STUDY NOROVIRUS IS THAT
THEY REFLECT THE HOST GENETIC
SUSCEPTIBILITY TO THE VIRUS. SO
AS EARLY AS THE '70s IT WAS
KNOWN A GENETIC SUSCEPTIBILITY
FACTOR FOR THIS VIRUS AND THAT
IS THE EXPRESSION OF FUNCTION OF
TRANSFERASE. PEOPLE HAVE
FUNCTION OF TRANSFERASE,
(INAUDIBLE) SUGARS IN MUCOSAL
SECRETIONS EPITHELIAL CELLS,
SECRET TORRS AND THEY ARE
SUSCEPTIBLE TO DIARRHEA AND
VOMITING BY MAJORITY OF THE
NOROVIRUSES. ON THE OTHER HAND
IF YOU NON-SECRETER YOU DO NOT
GET INFECTED YOU DON'T BECOME
ILL WITH MOST VIRUSES. T WHAT
WAS INTERESTING WAS WHEN WE MADE
THE ORGANOIDS WE MADE ORGANOIDS
FROM ENTOIDS FROM SECRETERS.
ONLY GOT INFECT WITH THE G2 4
DOMINANT NOROVIRUS. NOT --
SUGGESTING THESE ORGANOIDS ARE
BEHAVING JUST LIKE WHAT YOU
WOULD EXPECT THEM TO IN TERMS OF
RETAINING GENETIC SUSCEPTIBILITY
PROFILE. WE TOOK THIS ONE STEP
FURTHER TO SEE HOW IMPORTANT IS
THE EXPRESSION OF GLYCANS SO WE
BASICALLY TOOK ONE SECRETER
ORGANOID AND ONE SECRET CRETEER
LINE AND MODIFY EXPRESSION OF
SUGAR EXPRESSI EXPRESSION. SO HU
HAVE A WILD TYPE SECRETER LINE,
WE KNOCKED OUT AND NO LONGER SEE
APICAL EXPRESSION OF THE GLYCAN
SHOWN, HERE YOU HAVE
NON-SECRETER LINE, THIS ONE WE
KNOCK IN FUT 2 GLYCAN
EXPRESSION. IT CHANGES THE
SUSCEPTIBILITY PATTERN OF
INFECTION SO YOU GO FROM A
SUSCEPTIBLE LINE TO COMPLETELY
RESISTANT LINE BUT YOU GO FROM
RESISTANT LINE TO A SUSCEPTIBLE
LINE. ALL THIS INDICATING THAT
THIS IS A REALLY GOOD MODEL FOR
US TO STUDY HUMAN NOROVIRUS.
WHICH BASICALLY MEANS YOU CAN
THEN USE THIS MODEL AS A
PLATFORM PRE-CLINICAL PLATFORM
FOR ANTIVIRAL TESTING. WE
STARTED WITH A FAIRLY SIMPLISTIC
APPROACH. WE TAKE NOROVIRUS
POSITIVE SUSPENSIONS WE ADD DRUG
OR NO DRUG TO HIV MONOLAYERS ON
96 WELL PLACE AND ASK TWO
QUESTIONS. IS THE DRUG CYTOTOXIC
AND IS THERE REPLICATION. AND WE
ASSESS THIS BY PATHWAY AND RTPCR
ASSAYS. WE LOOK AT DIFFERENT
CLASSES OF DRUGS. SO WE LOOK AT
MEDICINAL CHEMISTRY APPROACHES
FOR PROTEASE INHIBITORS.
NOROVIRUS IS A PROBLEM IN
IMMUNOCOMPROMISED PATIENTS SO
THERE ARE CERTAIN DRUGS USED FOR
ANECDOTALLY THEY WORK IN SOME
CASES DON'T WORK IN SOME CASES
WE WANT TO TEST THOSE DRUGS. WE
WANT TO SCREEN ANTIVIRAL
LIBRARIES AND MOST IMPORTANTLY
EXCITING FOR US IS BECAUSE WE
NOW HAVE MODEL SYSTEM FOR THIS
VIRUS, WE ARE NOW IN BIOLOGY AND
SO BASED ON NEUROBIOLOGY
FINDINGS CAN YOU IDENTIFY
COMPOUNDS USED TO TREAT
NOROVIRUS AND I SHOW YOU THE
DATA FOR ALL THESE. GOING TO
SHOW YOU DATA WITH FOUR HIV HIE
LINES J 2 AND J 4 AND
GENETICALLY MODIFIED LINES FOR
KNOCK OUTLINE ON THE J 2
BACKBONE AND KNOCK IN LINE ON
THE J 4 AND WE CHOSE THESE LINES
BECAUSE THESE LINES REALLY
SUPPORT BETTER REPLICATION OF
MANY HUMAN NOROVIRUSES. WE WERE
LOOKING AT ASSAYS AND RT PCR.
WHEN WATERED DOING TRANSFERASE
WE HAVE AN INTERESTING
OBSERVATION. THIS IS WORK LED BY
A GRADUATE STUDENT IN THE GROUP,
MIRANDA NOTICED WHEN SHE GAVE
THESE DIFFERENT LINES SO FOUR
LINES HERE J 2 (INAUDIBLE) J 4
AND J 4 RAT THE BOTTOM, WHEN YOU
TAKE THESE LINES AND EITHER
LICENSE CONTROL OR TAKE THE --
IN OUR CASE DMSO OR A DRUG, I
WILL SHOW YOU IN A MINUTE, YOU
SEE THE STANDARD PROTOCOL WHICH
IS 30 MINUTES THE VALUES REALLY
AT LIMIT OF INFECTION SO 3 OR
HIGHER WHICH MEANS YOU DON'T GET
ACCURATE RESULTS WITH THOSE
VALUES. WE RECOMMEND YOU SHORTEN
THE TIME. SHE SHORTENED THE
TIME TO 20 MINUTES 10 MINUTES
AND FIVE MINUTES. IT IS ONLY AT
FIVE MINUTES YOU START TO SEE
SOME VALUES THAT ARE MORE LIKELY
TO BE WITHIN THE RANGE. WHEN YOU
USE THE (INAUDIBLE) TOXICITY THE
DATA IS STRIKING. I WANT TO SHOW
YOU DATA FROM TWO LINES. THE J 2
LINE WITH (INAUDIBLE) IF YOU
LOOK HERE AT THE 30 MINUTE TIME
POINT, DMSO LOOK LIKE THEY ARE
90% CYTOTOXIC WHICH WAS NOT THE
CASE WHEN YOU LOOK AT THE CELLS
UNDER THE MICROSCOPE. WHAT IS
INTERESTING ALSO IS THE TWO
LINES ARE VERY DIFFERENT FROM
EACH OTHER. SO THE J 4 FUT 2 FOR
EXAMPLE, LOOKS LIKE 30%
CYTOTOXIC, THE DRUG 80%
CYTOTOXIC. BUT IF YOU SHORT TN
ASSAY TO 5 MINUTE TIME. THERE IS
CYTOTOXICITY AND IF YOU DO
RELATIVE CYTOTOXICITY
CALCULATIONS THEY ARE GOING TO
BE QUITE DIFFERENT WITHIN
DIFFERENT TIME POINTS. SO THIS
GOT MIR DRAIN THINKING HOW DO
YOU MODIFY -- MIRANDA THINKING
ABOUT HOW TO MODIFY THE ASSAY
AND TAKE THIS THAT YOU USE FOR
THE ASSAY AND DILUTE THEM TO
FIGURE OUT WHERE YOU GET SOME
SORT OF LINEAR RANGE. ONCE AGAIN
THE DATA FROM FOUR LINES SHOWN
OVER HERE, 50 CALCULATIONS TO
FIND WHERE EXACTLY YOU HAVE END
POINTS AND YOU CAN SEE THE
DILUTION VARIES FOR THE
DIFFERENT LINES. SO NOW WHEN
YOU CHOOSE THIS GOOD DILUTION
AND ASSAY YOUR RESULTS ARE
DIFFERENT. SO ONCE AGAIN DATA
FOR J 2 AND JFUT 2 WHAT YOU SEE
HERE IS THAT FOR THE J 2 LINE
DMSO IS 50% CYTOTOXIC. THE DRUG
IS 30%. FOR THE J 2 IT IS 5% AND
20%. THIS IS WHAT WE SEE UNDER
THE MICROSCOPE. AND WE DON'T SEE
BIG DIFFERENCE BETWEEN THE
DIFFERENT TIME POINTS. ALL THIS
TO SUGGEST THE THAT MODIFYING
THE ASSAY BY DILUTING MAYBE
SOMETHING IMPORTANT TO CONSIDER.
IN ADDITION, MIRANDA TESTED
OTHER COMMERCIALLY AVAILABLE
ASSAYS, TO SEE SHOULD WE BE
GOING FOR OTHER ASSAYS INSTEAD
OF LDH ASSAYS. I SHOW YOU DATA
HERE OF THE RESULTS WHICH IS
USED IN IMMUNOCOMPROMISED
PATIENTS AGAIN SOMETIMES WORK,
DOESN'T WORK SOMETIMES. WHAT YOU
SEE HERE IS LOOK AT DMSO
NON-CYTOTOXIC CONCENTRATION OR
CYTOTOXIC CONCENTRATION, THERE
IS A BIG DIFFERENCE BETWEEN LDH
ASSAYS IN DARK BLUE COMPARED TO
ALL THE OTHER ASSAYS AND THAT
HOLDS TRUE FOR BOTH LINES.
IMPORTANTLY THAT'S JUST LOOKING
HERE AT THE J 4 FUT 2 LINE. THE
-- DOES SUGGEST 70% VIABLE.
UNDER THE MICROSCOPE OR FLOW
SIGH FOE METRICS IT IS 70% DEAD
SO ALL THIS IS TO TELL US THAT
WHEN YOU ARE USING MEDICINAL
ORGANOID CULTURES, WE MAY NEED
TO LOOK AT CYTOTOXICITY BY MORE
THAN ONE ASSAY AND WE NEED TO BE
VERY MINDFUL THAT THERE IS
VARIABILITY BETWEEN ENTROID
LINES. SO WE ARE TESTING
ANTIVIRALS AT THAT TIME SAME
TIME AND WE HAVE A PIPELINE FOR
ANTIVIRAL DESIGN AND DEVELOPMENT
IN TESTING. AS I MENTION WE CAN
DIVIDE WHAT WE DO IN A FEW
CATEGORIES AND I WILL SHOW YOU
EXAMPLESES OF ONES THAT WORK FOR
US. AS A VIROLOGIST YOU ARE
INTERESTED IN PROTEASE AND
POLYMERASE INHIBITORS. THE --
SHOWN ON THE BOTTOM HERE, SO
HERE I HAVE TO SHOW WHEN YOU
GAVE (INAUDIBLE) VIRUS NOROVIRUS
YOU EVENTUALLY VARY
SIGNIFICANTLY REDUCE G1
REPLICATION IN A HOST DEPENDENT
MANNER. AS I MENTIONED, WE ARE
LOOMING WITH BIOLOGY, THAT GIVES
OPPORTUNITIES TO THINK ABOUT
REPURPOSING DRUGS, WHAT WE HEARD
IN THE FIRST TALK. SO WHEN WE
PUBLISH THE SCIENCE PAPER AND
LOOK AT VIRUS REPLICATION, WE
NOTICE THE GLOBALLY DOMINANT G2
4 STRAIN GO TO ANDROIDS WITHOUT
A PROBLEM BUT OTHER VIRUSES FOR
EXAMPLE IN G2 3 STRAIN YOU
REQUIRE BILE OR BILE ACID FOR
REPLICATION. REPLICATION IN THE
PRESENCE OF BILE. WHICH LED US
TO ASK IF YOU USE A BILE ACID
STRAIN WHAT HAPPENS? WE ACTUALLY
SEE THE BILE ACID SEQUENCE WITH
CHOLESTEROL STY RA MEAN INHIBITS
NOROVIRUS. SIMILARLY WE LEARNED
THAT WHEN YOU USE BILE OR BILE
ACID IN THIS CASE BILE ACID YOU
HAVE RAPID INCREASE IN
REGENERATION IN ENTROID
CULTURES, HAPPENS WITHIN TEN
MINUTES. YES GCDCA AND RED
STAINING, AND THIS IS IMPORTANT
FOR THE REPLICATION OF THIS
PARTICULAR VIRUS, G2, 3 VIRUS.
SO WHAT HAPPENS IF YOU USE
SOMETHING WHICH INHIBIT THE
CERAMIDE FORMATION. THIS IS
MEDIATED BY MILLENASE, SO WHEN
YOU USE INDIVIDUAL LIKE AIM TRIP
LIEN WHICH IS USED AS AN
ANTIDEPRESSANT BUT ALSO ACE
INHIBITOR YOU CAN SEE IT IS DOSE
DEPENDENT REDUCTION IN NOROVIRUS
REPLICATION. IN ADDITION TO ALL
THESE TARGETED APPROACHES YOU
WANT TO LOOK AT LIBRARIES AND
SCREEN LIBRARIES ANTIVIRALS. AS
AN EXAMPLE HERE I SHOW YOU DATA
WHICH CAME OUT OF A GROUP IN
JAPAN, DATA SCREEN 3626
COMPOUNDS USING HUMAN INTESTINAL
ANDROIDS AND IDENTIFY ANTIVIRAL
WHICH IS NOT CYTOTOXIC AND HAS
AAN INHIBITORY EFFECT ON HUMAN
NOROVIRUS REPLICATION. SO DID
THIS WITH MULTIPLE LINES AND
IDENTIFIED IT CAN INHIB
MULTIPLE NOROVIRUSES. SO WHAT I
WANT YOU TO TAKE AWAY FROM ALL
THIS IS HUMAN INTESTINAL
ANDROIDS ARE EXCELLENT MODEL FOR
PREVIOUSLY NON-COMPATIBLE
VIRUSES AND IMPORTANTLY IN THE
CONTEXT OF ANTIVIRAL, IT IS A
GREAT PRE-CLINICAL TOOL FOR DRUG
TESTING. BUT I THINK IT IS
IMPORTANT TO BE COGNIZANT OF THE
FACT THAT ORGANOIDS REALLY MIMIC
THE HOST VARIABILITY THAT IS
SEEN IN RESPONSE TO DRUGS. SO IF
YOU ARE USING THIS ANTIVIRAL YOU
WILL NEED TO CONSIDER TESTING
MULTIPLE LINES, AND THAT BRINGS
THE QUESTION WHAT IS A GOOD
NUMBER TO USE. I DON'T THINK WE
REALLY HAVE A GOOD ANSWER AT
THIS POINT IN TIME. WE HAVE
LEARNED THROUGH OUR -- THAT IT
IS IMPORTANT TO EVALUATE
CYTOTOXICITY BY MULTIPLE
MEASURES AND YOU NEED TO
CONSIDER MODE OF ACTION IF YOU
KNOW IT FOR CHOOSING WHICH
CYTOTOXICITY TESTS YOU WILL USE.
YOU WILL NEED TO BE MINDFUL OF
VARIATION OVER TIME EVEN WITHIN
THE SAME LINE. WE TRY TO MAKE
THIS SAME NUMBER OF CELLS EVERY
TIME AND ALL THAT BUT STILL
VARIABILITY IN SOMETHING WE NEED
TO BE MINDFUL OF. WE ARE MOVING
TO USING 384 WELL PLATE ASSAYS
WHERE YOU CAN TEST MULTIPLE
ORGANOIDS MULTIPLE NOROVIRUS
STRAINS AND I THINK THAT IS
REALLY WHERE AT LEAST FOR THE
NOROVIRUS DRUG TESTING THE NEXT
STAGE WHERE YOU CAN USE TO COME
UP WITH COMPOUNDS THAT ARE
AFFECTIVE. FINALLY, BIG PICTURE
PERSPECTIVE. ALL THE LINES WE
WORK WITH NOW ARE EPITHELIAL
CELLS ONLY BUT OF COURSE THE
FACTORS ARE MORE COMPLEX THAN
THAT AND WE ARE ESTABLISHING
CO-CULTURES WITH CELLS AND
ORGANOIDS AND PLATFORMS WHERE
YOU CAN MIMIC THE INTESTINAL
GRADIENT AND MICROBIOME FOR
CULTURE STUDIES. ALL THESE WILL
BEEN USEFUL FOR DRUG TESTING IN
THE FUTURE. SO WITH THAT I WANT
TO THANK EVERYBODY WHO HAS BEEN
PART OF THIS WORK. USE OF
ENTROIDS TO STUDY NOROVIRUSES,
THE WORK I SHOW YOU CAME FROM
RANDALL LOUIS A GRADUATE
STUDENT, SHE DID THE (INAUDIBLE)
WORK, BIOLOGY BASED DRUGS BY
VICTORIA AND THE ANTIVIRAL
LIBRARY STREAMING PAPER FROM
JAPAN. THANK YOU FOR YOUR
ATTENTION AND I LOOK FORWARD TO
YOUR QUESTIONS IN THE DISCUSSION
SESSION. THANK YOU.
>> THANK YOU SO MUCH, DR.
RAMANI. EXCELLENT PRESENTATION
AND ABSOLUTELY LOOK FORWARD TO
DISCUSSION LATER THIS AFTERNOON
>> THANK YOU.
>> LOST COUNT I THINK WE ARE AT
THE FOURTH SPEAKER OF THE
SESSION SO BEFORE OUR FIRST
BREAK. I WOULD LIKE TO INTRODUCE
DR. SARA CHERRY. DR. CHERRY IS A
PROFESSOR IN THE DEPARTMENT OF
PATHOLOGY AND LABORATORY
MEDICINE AT THE UNIVERSITY OF
PENNSYLVANIA AND IS A SCIENTIFIC
DIRECTOR OF THE HIGH LIEU PUT
SCREENING CORE AND DIRECTOR OF
THE CHEMOGENOMIC DISCOVERY
PROGRAM IN THE SCHOOL OF
MEDICINE THERE. SHE OBTAINED HER
BACHELOR'S DEGREE WORKING WITH
DR. PETER SCHULTZ AT BERKELEY,
HER Ph.D. WITH DAVID BALTIMORE
AT MIT AND POST-DOCTORAL
FELLOWSHIP WITH DR. PERRYMAN.
UPON STARTING HER LABORATORY AT
UPENN SHE HAS APPLIED HIGH LIEU
PUT SCREENING TECHNOLOGIES TO
KISS COVER MECHANISMS WHICH
VIRAL PATHOGENS HIJACK CELLULAR
MACHINERY EVADING DEFERENCES.
SHE IDENTIFIED INNATE IMMUNITY
AND CELLULAR INTERACTIONS
BETWEEN VIRUSES AND CELLS
COMPARING AND CONTRASTING VIRAL
FAMILIES. MORE RECENTLY SHE'S
UNCOVERED NEW INSIGHT TO THE
INNER PLAY BETWEEN METABOLIC
REGULATION, THE MICROBIOTA AND
IMMUNE DEFENSE. GIVEN THE
PANDEMIC HER LABORATORY HAS
APPLIED HER SCREENING PLATFORM
TO STUDY THE EMERGING CORONA
VIRUS SARS COV-2 IDENTIFY NEW
ANTIVIRALS, ACTIVE IN THE
RESPIRATORY TRACT. SHE UNCOVERED
ANTIVIRALS WITH DISTINCT
MECHANISMS OF ACTION INCLUDING
INNATE IMMUNE MODULATORS DIRECT
ACTING ANTIVIRALS AND HOST
DIRECTED ANTIVIRALS. BY
COMPARING ANTIVIRALS ACROSS
RELATED AND UNRELATED VIRUSES,
SHE IDENTIFIED SYNERGIES BETWEEN
ANTIVIRALS THAT MAY BE LEVERAGED
FOR COMBINATION THERAPIES IN THE
FUTURE. THE TITLE OF DR.
CHERRY'S TALK IS ANTIVIRAL
SCREENING PIPELINE AND LESSONS
LEARNED. OVER TO YOU, DR.
CHERRY.
>> THANK YOU VERY MUCH, ANN,
THANKS TO ORGANIZERS ALLOWING ME
TO PRESENCE THE WORK WE HAVE
BEEN DOING TO DEVELOP A PIPELINE
TO ALLOW US TO IDENTIFY NEW
ANTIVIRALS IN ORDER TO MOVE
THERAPEUTICS FORWARD POTENTIALLY
CLINICALLY. SO AS WE ALL KNOW,
WE HAVE HAD MANY EMERGING AND
RE-EMERGING VIRUSES
HISTORICALLY, THIS IS A IMAGE
FROM THE GATES FOUNDATION THAT
CAPTURES MUCH OF OUR MORTALITY
PRIOR TO THE -- TO 2020 WHERE
MOSQUITOES AND MOSQUITO BORNE
ILLNESSES LEAD TO LARGE BURDEN
AND MY LAB HISTORICALLY STUDIED
THESE ARTHEROPOD VIRUSES. ALSO
KNOW THE WORLD IS GETTING
SMALLER MORE INTERACTIONS AND
MORE LIKELIHOOD OF NEW VIRUSES
ENTER INTO POPULATIONS AND OF
COURSE THE LAST FEW YEARS
THERE'S BEEN UNBELIEVABLE BURDEN
OF INFECTION AND MORTALITY DUE
TO COVID-19. SO WHAT WE ARE --
WHAT WE THINK, MANY HERE IN THIS
AUDIENCE THINK IS WE NEED MORE
ANTIVIRALS AGAIN SARS COV-2 BUT
ALSO ALL THE OTHER EMERGING AND
RE-EMERGING VIRUSES WHICH WE
HAVE VERY FEW ANTIVIRALS. SO WE
THINK OF ANTIVIRALS IS THREE
MAJOR CLASSES. THERE'S THE
DIRECT ACTING ABOUT VIRALS WHICH
ARE LARGEST CATEGORY THAT BLOCK
VIRUS ITSELF, VIRAL ENZYME,
VIRAL ACTIVITY. THERE IS HOST
DIRECTED AB VIRALS REQUIRING
MACHINERY TO REPLICATE. YOUR
TARGET MACHINERY TO BLOCK
INFECTION. WE HAVE EXAMPLES FOR
ENTRY INHIBITORS. ANOTHER CLASS
OF ANTIVIRALS ARE THE ONES THAT
CAN MODULATE THE IMMUNE RESPONSE
AND I WILL TELL YOU TODAY ABOUT
ANTIVIRALS THAT WE HAVE BEEN
WORKING ON FROM ALL LEE
CATEGORIES. -- THREE CATEGORIES.
THE PIPELINE WE HAVE BEEN TRYING
TO IMPLEMENT IN SARS AND OTHER
VIRUSES IS THE FOLLOWING. SO WE
USE HIGH THROUGH PUT SCREENING
AND 384 WELL PLACE WHERE WE CAN
ADD CELLS THAT ADD DRUGS FROM
DRUG LIBRARIES USING ROBOTICS.
AND THEN WE CAN INFECT VIRUSES
SUCH AS SARS COV-2 AND OUR ASSAY
OF CHOICE USES MICROSCOPY, WE
HAVE AUTOMATED IMAGE ANALYSIS
THAT ALLOWS US TO LOOK AT WILD
TYPE VIRUSES STAINED WITH
ANTIBODIES AND ALLOWS US TO ALSO
LOOK AT CELL VIABILITY IN THE
SAME WELL. ALLOWING US TO FIND
THINGS THAT BLOCK INFECTION
WITHOUT CYTOTOXICITY UP FRONT IN
OUR SCREENS. WE GO ON TO
VALIDATE THE HITS FROM THE
PRIMARY SCREENS USING DOSE
RESPONSE STUDIES IN VARIETY OF
CELL TYPES AS WELL AS ORTHOGONAL
ASSAYS TO LOOK AT THE EFFICACY
BECAUSE WE AND MANY OTHERS THINK
THAT INHIBITORS HAVE TO BE
POTENT AND BLOCK INFECTION BY
MORE THAN A LOG TO HAVE
PHENOTYPES IN VIVO. WE HAVE BEEN
INCORN RATING PRIMARY CELL
MODELS INTO OUR PIPELINE AT THIS
POINT TO TRIAGE THOSE DRUGS THAT
MIGHT BE ACTIVE IN CULTURED CELL
LINES AND NOT IN VIVO. SO WE
HAVE BEEN USING LIQUID INTERFACE
CULTURES WHICH HAVE COME UP A
NUMBER OF TIMES WHERE THESE ARE
FULLY DIFFERENTIATED SILLIATED,
PRODUCE MUCOUS, AND WE FOUND FOR
SARS COV-2 THEY HAVE TO BE
CULTURES AND MORE THAN 8 DAYS
OLD. WE ARE USERS OF THIS
TECHNOLOGY NOT DEVELOPERS OF
THIS TECHNOLOGY AND SO WE HAVE
BEEN WORKING WITH NUMBER OF
COMPANIES TO UTILIZE THESE IN A
SYSTEMATIC WAY TO TEST SMALL
MOLECULES TO GET LIQUID
INTERFACE CULTURES FROM NASAL
EPITHELIUM, AND THE BRONCHIAL
EPITHELIUM FROM MATT TECH. IN
ADDITION AT PENN THERE IS A CORE
THAT PRODUCE HUMAN IPS DERIVED
CELLS, WE HAVE BEEN USING THIS
AS WELL. SO ONCE WE FIND DR.
DRUGS ACTIVE IN PRIMARY MODELS
TO DO MECHANISM STUDIES TO
UNDERSTAND HOW THEY MIGHT BE
ACTING, AS WELL AS TESTING THEM
ACROSS DIFFERENT VARIANTS AND
CORONA VIRUSES TO IDENTIFY
ANTIVIRALS THAT HAVE SOME
BREADTH. IN MANY ADDITION AS
MANY OF YOU KNOW THERE IS
CONCERN ABOUT MONOTHERAPY
BECAUSE VIRUSES CAN MUTATE HENCE
VARIANTS AND INCORPORATE
COMBINATION TESTING IN YOU ARE
PIPELINE TO SEE IF COMBINATIONS
OF DRUGS ARE ACTIVE AS WELL.
COLLABORATION WITH A NUMBER OF
OUR COLLEAGUES WE HAVE MOVED
SOME OF THESE TESTS IN TO
PRE-CLINICAL MODELS, IN SMALL
ANIMALS, SUCH AS MICE WITH HOPES
THAT ONCE WE PASS ALL THESE
HURDLES WE CAN USE SOME OF THESE
DRUGS INTO CLINICAL TRIAL. SO
TO START WITH HIGH THROUGH PUT
SCREENS, WE COME UP WITH NUMBER
OF LIBRARIES THAT WE HAVE BEEN
SCREENING. AS MANY AS YOU HAVE
HEARD ABOUT THE FASTEST WAY TO
GET MOVING IS TAKING DRUGS THAT
EXIST, THAT'S WHAT WE DID AS
WELL. WE SCREENED 20,000 DRUGS
ARE FROM A VARIETY OF
REPURPOSING LIBRARIES INCLUDING
WORKING WITH NCATS TO LEVERAGE
THEIR LIBRARIES AS WELL. AS WITH
GATES FOUNDATION TO TEST THE
REFRAME LIBRARY. WE ARE ARE ALSO
INCORPORATING DIVERSITY
LIBRARIES BUT I'M NOT GOING TO
HAVE TIME TO TALK ABOUT THOSE
TODAY. SO THE DRUG LIBRARY IS
ONE FACET OF HIGH THROUGH PUT
SCREEN BUT OTHER IS USED IN CELL
TYPES. SO AS YOU HAVE HEARD
EARLIER TODAY, IT IS CLEAR THIS
VIRUS CAN USE MULTIPLE MODES OF
ENTRY. EARLIER STUDIES USING HH
7 CELLS AND VIRILE CELLS AN
NON-RESPIRATORY CELLS SHOWED THE
VIRUS CAN ENTER CELLS THROUGH
ENDOCYTIC MECHANISM DEPENDENT ON
THE PH REQUIRED TO ACTIVATE
CAPSAICIN PROTEASES SO CAPSAICIN
INHIBITORS ARE ACTIVITY AS WELL
AS HYDROXYCHLOROQUINE AND
INHIBITOR INDICATIONS ARE ACTIVE
IN NON-RESPIRATORY CELLS. IN
CONTRAST WE KNOW ACTIVE IN
CERTAIN CONTEXT, TO TRIGGER
ENTRY IS NOT ACTIVE IN
ACTIVATORS ARE NOT ACTIVE IN
NON-RESPIRATORY CELLS BECAUSE
THEY DON'T EXPRESS TMPRSS 2. WE
HAVE SCREENED MANY CELL LINES TO
FIND RESPIRATORY CELL LINE
PERMISSIVE TO INFECTION
ENDOGENOUSLY AND WE CAME UPON
CELLS ROBUSTLY INFECTED IN CELL
CULTURE MODIFICATION. SO IN
THESE RESPIRATORY CELLS THE
CATHEPSIN CELLS ARE DEAD AND
TMPRSS 2 INHER TOES ARE ACTIVE
AND INHIBIT INFECTION AND THEY
ARE SO NON-TOXIC THE VIRUS WITH
GROW -- CELLS WITH GROW BETTER
IN THE PRESENCE OF THIS
INHIBITOR. I WANT TO POINT OUT
THE CELLS MANY PEOPLE USE TO
MODEL SARS COV-2 USES ENDOCYTIC
ENTRY AND NOT THE TMPRSS 2, THEY
ARE A BETTER MODEL FOR INDEED
ALL THE PRIMARY RESPIRATORY
CELLS THAT WE HAVE TESTED USE
TMPRSS 2 FOR ENTRY. SO IN THE
IPS DERIVED CELLS INTERFACE
CULTURES WE CAN BLOT INFECTION
AS MEASURED BY -- BLOCK
INFECTION MEASURED BY RTPCR.
ALSO IN OUR HAND THE VARIANTS
ARE DEPENDENT ON TMPRS 2 FOR
INFECTION, THESE ARE ICP USING
TMPRR 2 INHIBITOR TO INHIBIT ALL
THE VARIANTS WE TESTED TO DATE.
THIS LED US TO USE THE CELLS FOR
THE CELL LINE OF CHOICE FOR OUR
INITIAL SCREENS. SO WHAT WE
FOUND EARLY ON IN THESE CELLS
AND IN OTHER RESPIRATORY MODELS
IS THAT THE INNATE IMMUNE SYSTEM
WHICH IS IMPORTANT TO INITIALLY
CONTROL INFECTION IS ACTUALLY
DELAYED ACTIVATION OF
INTERFERONS AND INNATE IMMUNITY
THE DELAYED UPON INFECTION WITH
SARS COV-2. SO WHILE MANY
VIRUSES INCLUDING INFLUENZA AND
RSV ACTIVATE EARLY TIME POINTS
POST INFECTION WE AND OTHERS
FOUND THAT SARS COV-2 ONLY
ACTIVATES INTERFERON IN THE
CELLS LATE TIME POINT POST
INFECTION. HOWEVER WHILE DELAYED
INDUCTION WE ALSO FOUND THAT
INTERFERONS CAN ROBUSTLY BLOCK
INFECTION IF TREATED BEFORE
INFECTION. YOU CAN SEE HERE WITH
INTERFERON BETA OR LAMBDA, YOU
CAN ROBUSTLY BLOCK INFECTION
MANY THESE 3 CELLS. GIVEN THAT
THE VIRUS SENSITIVE INTERFERONS
WE WERE WONDERING IF THERE'S
OTHER INNATE LIGANDS THAT MIGHT
BE ABLE TO HAVE ACTIVITY TO
BLOCK INFECTION. SO CREATE AD
LIBRARY WHAT ARE CALLED PAMS OR
PATHOGEN ASSOCIATED PATTERN TO
INNATE AGONISTS AND SCREEN FOR
ACTIVITY IN CAL 3 CELLS. SO WHAT
WE FOUND IS TWO OF THE LIGANDS
CYCLIC NUCLEOTIDES WERE QUITE
POTENT TO BLOCKING SARS COV-2
INFECTION IN CALU 3 CELLS. SO
THIS IS DOSE RESPONSE STUDIES WE
CAN SHOW THAT WE HAVE NICE DOSE
DEPENDENT ACTIVITY AND BY QPCR
WE CAN SEE WE CAN BLOCK
INFECTION. HOWEVER, EC 50s
HERE ARE ACTIVE BUT VERY HIGH
CONCENTRATIONS AND THAT IS
BECAUSE THESE DINUCLEOTIDES ARE
NOT CELL PERMEABLE. BECAUSE OF
THIS NUMBER OF PHARMA COMPANIES
DEVELOPED SMALL MOLECULES STAIN
AGONISTS SO WE RETAINED ONE
AGONIST AND WE FOUND THAT IT IS
POTENTLY ACTIVE IN CALU 3 CELLS
TO BLOCK INFECTION AND BLOCK
VIRAL REPLICATION BY THREE
ORDERS OF MAGNITUDE. NEXT WE
MOVE TO THE AIR LIQUID INTERFACE
CULTURE DESCRIBED IN THE
PIPELINE AND WE WORK WITH EMILY
AND MARK IN THESE STUDIES
OBTAINING CULTURES FROM MATT
TECH AND INFECT WITH SARS COV-2,
AND WE CAN REALLY POTENTLY BLOCK
VIRAL REPLICATION SHOWN BY
CONFOCAL MICROSCOPY IN THE
CENTER OR BY RTPCR ON THE RIGHT.
WHERE THESE -- THIS SMALL
MOLECULE LIGAND IS ACTIVE
AGAINST SARS COV-2. NEXT MOVE TO
ANIMAL MODEL COLLABORATION WITH
(INAUDIBLE) LAB WE INITIALLY USE
ANCESTRAL STRAIN OF SARS COV-2
AND INFECTED THE HUMAN
TRANSGENIC LINE CALLED K 18. IN
THIS CASE WE INTRANASALLY
DELIVERED THE LIGAND AND THEN
CHALLENGED INTRANASALLY WITH
SARS COV-2. WHAT WE CAN SEE IS
WE CAN PROTECT THE ANIMALS FROM
WEIGHT LOSS AND MORTALITY DUE TO
INFECTION, REALLY ROBUST
INHIBITION OF VIRAL REPLICATION
IN THE LUNGS. WE NEXT WANT TO
MOVE TO MORE RELEVANT MODEL
INFECTION SO AS MANY OF YOU
PROBABLY KNOW THE SOUTH AFRICAN
VARIANT INFECTS WILD TYPE, YOU
CAN USE THE MOUSE ACE 2 RECEPTOR
SO WE CAN USE THE SOUTH AFRICAN
VARIANT CHALLENGE WITH 129 WILD
TYPE MICE, IN THIS CASE THE
INFECTION IS LESS LETHAL SO WE
CAN ACTUALLY TREAT THE ANIMALS
WITH THIS MITIGATING WEIGHT LOSS
AND ATTENUATING VIRAL
REPLICATION, SUGGESTING WE CAN
GO FROM IN VITRO ASSAY ALL THE
WAY THROUGH PRE-CLINICAL MODELS
AND SHOW REALLY HIGH ACTIVITY.
AS I SAID WE HAVE SCREENED
20,000 DRUGS AT THIS POINT.
INCLUDING THOSE LIGANDS BUT MORE
CLASSICAL LIBRARIES THAT I
DESCRIBED EARLIER. SO FROM THESE
20,000 DRUGS WE THEN WENT ON TO
DO DOSE DEPENDENT STUDIES AND WE
VALIDATED 150 DRUGS THAT HAD
INDEXES AND QUITE ACTIVE. SO THE
TWO CATEGORIES THAT I WANT TO
MENTION NOW ARE THE NUCLEOSIDE
ANALOGS AND METABOLIC
INHIBITORS. WE IDENTIFIED 16
NUCLEOSIDES THAT HAD ANTIVIRAL
ACTIVITY AND NUCLEOSIDE ANALOGS
ARE POTENT ANTIVIRALS, MANY CAN
BE INCORPORATED BY THE
POLYMERASE MEETING TO AFFECTING
THE VIRAL RNA REPLICATION. SO
SHOWN ON THE RIGHT HERE ARE THE
IC 50 CURVES OF CALU 3 TREAT
WITH REMDESIVIR, TO GO MEAN,
WHERE YOU CAN SEE THAT THEY HAVE
POTENT ACTIVITY IN THESE CELLS.
OF COURSE WE THEN TESTED THESE
SMALL MOLECULES IN MORE RELEVANT
CELL LINES SO WE TRIED -- WE
USED THE DERIVED 2 CELLS,
BRONCHIAL EPITHELIAL LINES FROM
MATTEK AND NASAL EPITHELIAL,
THIS IS LOOKING AT FOUR OF THOSE
ANALOGS WHERE YOU CAN SEE THAT
THEY ALL HAVE REMDESIVIR
ACTIVITY AGAINST ALL THESE LINES
AS DOES (INAUDIBLE) BUT THERE IS
MORE STRIKING DIFFERENCES
BETWEEN THE MAT TEK AND AIR
LIQUID INTERFACE CULTURE THAN
THE IA 2 AND CALU 3 TELLS WHERE
GUANINE HAVE NO ACTIVITY IN THE
AIR LIQUID INTERFACE MODELS,
AGAIN REALLY SUGGESTING THAT
GOING THROUGH THESE DIFFERENT
STEPS TO TRIAGE MOLECULES BEFORE
WE MOVE FORWARD IS REALLY
IMPORTANT. THE OTHER CLASS IN
THE PIE CHART THERE METABOLIC
INHIBITORS AND WE IDENTIFIED A
NUMBER OF BIOSYNTHESIS AND PIR
RENNE BIOSYNTHESIS INHIBITORS SO
THESE ARE SMALL MOLECULES
BLOCKING THE HOST WHILE BLOCKING
THE BIOSYNTHESIS, THE VIRUS ALSO
NEEDS TO USE FOR IT TO UNDERGO
RNA REPLICATION. SO WE WERE
REALLY INTERESTED IN WHETHER OR
NOT WE CAN DO COMBINATION
THERAPIES, TO IDENTIFY SYNERGIES
BETWEEN DIFFERENT CLASSES OF
SMALL MOLECULES, TO POTENTIALLY
ALLOW FOR LOWER DOSES IN HIGHER
BARRIER TO RESISTANCE. SO WE
COMBINED THE PRIM DEAN
BIOSYNTHESIS INHIBITOR
BIOAVAILABLE WITH ORALLY
BIOAVAILABLE AND USING TWO BY
TWO MATRIX TESTING DOSE RESPONSE
BY DOSE RESPONSE, WE ARE ABLE TO
SHOW VERY POTENT SYNERGY AS
MEASURED BY BLISS ANALYSIS
BETWEEN THE TWO DRUGS SUGGESTING
THAT THERE MAYBE A REAL BENEFIT
TO COMBINING THESE TWO DIFFERENT
SMALL MOLECULE MECHANISMS IN
VIVO. ALSO ASK WHETHER OR NOT
THIS IS SPECIFIC TO (INAUDIBLE)
OR ALSO TRUE WITH REMDESIVIR,
ANOTHER NUCLEOSIDE ANALOG AND WE
WERE ABLE TO SHOW SYNERGY WITH
THE COMBINATION OF REMDESIVIR
AND BRECINAR ALSO SUGGESTING
THAT BY COMBINING NUCLEOSIDE
ANALOG WITH A BIOSYNTHETIC BLOCK
WE MAY SHOW REALLY GOOD ACTIVITY
IN VIVO. EARLIER I WANT TO POINT
OUT WE DID THESE WITH PAXLOVID
AND THE PROTEASE INHIBITOR AND
WHEN WE COMBINE THAT WE SEE
ACTIVITY NOT SYNERGY SUGGESTING
A GOOD COMBINATION TO USE FOR
POTENTIALLY MOVING FORWARD. SO
THEN WE MOVED TO THE AIR LIQUID
INTERFACE MODELS, AND WITH
EPITHELIUM WE VALIDATED DOSING
OF THE CULTURES SO THIS
SOMETHING THAT MAYBE HASN'T COME
UP OFTEN BUT IT IS IMPORTANT TO
MAKE SURE THAT THE CULTURES ARE
HEALTHY AND ACTIVE WHEN TREATING
WITH THESE SMALL MOLECULES SO
THEY RUN ASSAYS INCLUDING TIERS
FREQUENCY AND TOXICITY AND
IDENTIFY CONCENTRATIONS THAT ARE
NOT AFFECTING THE VIABILITY AND
PERMEABILITY OF THE AIR LIQUID
INTERFACE CULTURES. SO THEN WE
TESTED THESE COMBINATIONS USING
THESE AIR LIQUID INTERFACE
CULTURES IN BOTH THE NASAL AND
BRONCHIAL MODELS. THIS IS A
LITTLE BUSY BUT YOU CAN SEE THAT
WHEN WE TREAT WITH THE 2801 WE
SEE A DOSE DEPENDENT CHANGE IN
REPLICATION WITH HIGHER DOSE WE
HAVE DECREASE IN PATIENT
COMPARED TO DIFFERENT DOSES AND
THAT IF WE THEN COMBINE LOWER
DOSE OF PIRAVIR WITH TWO
DIFFERENT BASSINET SINCE HUNDRED
TORRS WE SEE A VAST --
BIOSYNTHESIS, WE SEE REPLICATION
COMPARED TO DRUGS ALONE IN NASAL
CULTURES AS WELL AS BRONCHIAL
CULTURES SUGGESTING THAT AGAIN
THIS COMBINATION MIGHT BE
BENEFICIAL IN VIVO. THEN WE
COLLABORATE WITH MATT FREEMAN'S
LAB TESTED IN VIVO MODEL USING
THE SOUTH AFRICAN VARIANT WITH
NASAL CHALLENGE AND AGAIN YOU
CAN SEE HERE IF WE USE DIFFERENT
DOSES OF PIROVIR WE CAN HAVE
DOSE DEPENDENT REDUCTION IN
VIRAL REPLICATION AND WHEN WE
ADD BRECLINAR TO THE COMBINATION
THERAPY WE CAN REDUCE
REPLICATION AT EVERY
CONCENTRATION OF EIKD 2801 SO
INCREASE REDUCTION IN TITERS
COMPARED TO EITHER OF THE DRUGSS
ALONALONE. WE WERE EXCITED TO SE
NOT ONLY DO WE HAVE REDUCTION IN
TITERS WE SAW REDUCTION IN
INFLAMMATION, SO THIS IS FROM
THE LUNGS OF THE ANIMALS, EITHER
NOT TREATED WITH DRUGS WHERE YOU
CAN SEE THIS TERRIBLE
INFLAMMATION AND AIRWAYS LOOK
TERRIBLE IF WE TREAT WITH
PIROVIR ALONE YOU CAN SEE NOT A
COMPLETE RESTORATION OF THE
LUNGS AS YOU CAN SEE IN THE
UNAFFECTED ANIMALS BUT WE
COMBINE BREQLINAR WE CAN RESTORE
THE ARCHITECTURE BACK TO WILD
TYPE SUGGESTING THAT WE HAVE
BOTH DECREASE IN TITTEDDERS AND
INFLAMMATION COMBINATION. SO
MOVING FORWARD WE ARE SCREENING
MORE LIBRARIES DIVERSITY
LIBRARIES TO IDENTIFY NEW MATTER
AND POTENTIALLY NEW DIRECT
ACTING ANTIVIRALS THEN MOVING
PIPELINE TESTING AGAINST DIVERSE
CORONA VIRUSES IN PRIMARY CELL
MODELS AND MECHANISM OF ACTION
STUDIES IN ORDER TO BRING IT
BACK TO ANIMAL MODELS AND
POTENTIALLY -- MY LAB IS
STUDYING EMERGING AND
RE-EMERGING VIRUSES, LARGE
ARTHEROPOD BORN SO WE ARE
ACTUALLY NOW USING THIS KIND OF
MODEL TO DEVELOP A PIPELINE TO
SCREEN ALL MAJOR FAMILIES OF RNA
VIRUSES THAT ARE TRANSMITTED BY
DIVERSE ANIMALS TO HUMANS IN
NATURE. ALONG WITH THE
DEVELOPMENT OF THESE ANTIVIRAL
SCREENINGS STRATEGIES WE ARE
TRYING TO DEVELOP MORE WORK WITH
OTHERS TO OBTAIN MORE
SOPHISTICATED 3D MODELS OF
DIFFERENT CELL TYPES THESE
VIRUSES INFECT IN HUMAN SARS
PATHOGENS. I WOULD LIKE TO PACK
KNOW LEDGE MY LAB, AND THE HIGH
THROUGH PUT SCREENING CORE WHICH
REALLY FACILITATED ALL THESE
STUDIES OVER THE LAST TWO YEARS.
ANIMAL STUDIES COLLABORATORS AS
WELL AS FOLKS AT NCATS,
MEDICINES THE GATES FOUNDATION,
THAT IS FACILITATED STUDIES AS
WELL AS INDIVIDUALS AND DEVELOP
THESE MODELS THAT ALLOWED US TO
RAPIDLY IMPLEMENT THEM FOR OUR
ANTIVIRAL STUDIES. WITH THAT,
THANK YOU.
>> EXCELLENT. THANK YOU SO MUCH,
DR. CHERRY, FOR A FANTASTIC TALK
AND TO THE PRIOR SPEAKERS
EARLIER IN THIS SESSION. SO WE
HAVE TWO MORE SPEAKERS TO GO AND
THEN THE QUESTION AND ANSWER
PERIOD AFTER LUNCH. BUT LIKE TO
TAKE A BRIEF BREAK, POLLS FOR
EATING INTO THE TIME A LITTLE
BIT BUT LIKE TO KEEP TO THE
SCHEDULE PLEASE AND RETURN FROM
BRIEF LUNCH BREAK AT 1:10 LOCAL
TIME. SO TEN AFTER HOUR WE WOULD
LIKE TO COME BACK AND FINISH OUT
SESSION 2. TWO MORE SPEAKERS AND
GET O THE QUESTIONS PERIOD. SO
ALSO BE PLEASE THINKING O YOUR
QUESTIONS AND FEEL FREE TO ADD
THEM TO THE Q&A CHAT BOX SO WE
WILL BE SURE TO COVER THEM
DURING THAT DISCUSSION. THANKS
AGAIN. SEE EVERYONE BACK AT 10
AFTER THE HOUR.
>>I THIS I IT'S TIME TO RESUME
OUR SESSION 2 DISSECTIONS. SO
THANK YOU AGAIN TO THE PRIOR
FOUR SPEAKERS. I NOW LIKE TO
INTRODUCE OUR FIFTH SPEAKER FOR
THE SESSION, DR. EMILY LEE. DR.
LEE IS A STAFF SCIENTIST AND
FUNCTIONAL GROUP LEAD AT NCATS
NIH. DR. LEE JOINED THE EARLY
TRANSLATION BRANCH IN THE
DIVISION OF PRE-CLINICAL
INNOVATION AT NCATS IN 2019 AS
BIOLOGIST. DRAWING ON HER
DIVERSE BACKGROUND IN VIROLOGY
HIGH THROUGH PUT DRUG SCREENING
AND COMPLEX IN VITRO MODELS DR.
LEE LEADS ADVANCE MODELS AND
CELL DISCOVERY ASSAY GROUP
WITHIN THE ANTIVIRAL PROGRAM FOR
PANDEMICS. AS A POST-DOCTORAL
FELLOW IN THE THERAPEUTICS
BRANCH AT NCATS DR. LEE
DEVELOPED HIGH LIEU PUT ASSAYS
FOR ANTIVIRAL SCREENING AS WELL
AS RARE GENETIC DISEASES. DR.
LEE EARNED HER DOCTORAL DEGREE
IN CELL AND MOLECULAR BIOLOGY
FROM THE FLORIDA STATE
UNIVERSITY WHERE SHE WORKED WITH
HENLEY STUDYING HOST PATHOGEN
INTERACTIONS AND IDENTIFYING
POTENTIAL ANTIVIRAL COMPOUNDS
WITH A FOCUS ON FLAVI VIRUSES.
HE CURRENT RESEARCH FOCUSES ON
DEVELOPING AND CHARACTERIZING
PHYSIOLOGICALLY RELEVANT CELL
BASED AND TISSUE ENGINEERED
PLATFORMS FOR VIRAL DISEASE
MODELING AND ANTIVIRAL DRUG
DISCOVERY. SHE AND HER TEAM ARE
CURRENTLY ENGAGED IN
COLLABORATIVE PROJECTS
PERTAINING TO HIGH IMPACT BLS 2
AND 3 VIRAL INFECTIONS INCLUDING
SARS COV-2. OF DR. LEE'S TEAM
WORKS CLOSE WITH MEMBERS OF THE
3D TISSUE BIBIOPRINTEDDING LAB
AT EARLY TRANSLATION BRANCH AT N
CATS AS WELL AS ACADEMIC
GOVERNMENT AND INDUSTRY
PARTNERS. SHE WILL BE SPEAKING
ON DEVELOPING MODELS FOR VIRAL
INFECTION AND DISEASE TO
INCORPORATE INTO THE DRUG
SCREENING PIPELINE. OVER TO YOU,
DR. LEE.
>> THANKS, DR. EAKIN. GOING TO
PULL UP MY SCREEN. CAN YOU SEE
THE CORRECT SCREEN?
>> YES. LOOKS GREAT.
>> GREAT. LET ME PULL THIS UP.
THANK YOU FOR INTRODUCTION.
THANK YOU TO EVERYONE ATTENDING
THE ATM WORKSHOP TODAY. I'M
HONORED TO SPEAK WITH Y'ALL
ABOUT SOME OF THE STRATEGIES WE
ARE TAKING ON 3D COMPLEX IN VIVO
MODELS.S. FOR INFECTIONS IN
DISEASE WITH THE GOAL OF
ENHANCING THE DRUG DEVELOPMENT
PIPELINE. I WILL START WITH WHO
WE R. THE IC AT THE NIH I WORK
IN IS NATIONAL CENTER FOR
TRANSLATION SCIENCES OR NCATS,
I'M A STAFF SCIENTIST MANY THE
3D LAB AND ALSO LEAD A SMALL
TEAM WITHIN THE SMALL TEAM APP.
MY TEAM IS LUCKY WE ARE A MIX OF
VIROLOGISTS, HIGH THROUGH PUT
SCIENTISTS AND TISSUE ENGINEERS
NESTLED TO LARGER GROUP OF DRUG
DISCOVERY AND HIGH THROUGH PUT
ASSAY SCIENTISTS MEDICINAL
CHEMIST, NAI TEAM AS WELL AS BY
LARGE GROUP OF TISSUE ENGINEERS
IN THE 3D TISSUE BIOPRINTING LAB
WE WORK AND COLLABORATE WITH
EVERY DAY ON DEVELOPING
UTILIZING TISSUE MODELS FOR
VIRAL INFECTION SO THE WORK WE
DO WOULDN'T BE POSSIBLE WITHOUT
THIS DIVERSE RANGE OF EXPERTISE
WORKING TOGETHER. SO TO
UNDERSTAND WHAT AND WHY WE ARE
TALKING ABOUT WHAT WE ARE, IT IS
IMPORTANT TO KNOW THE GOALS OF
OUR TEAM. WE ARE PASSED
DEVELOPING 2D ANTIVIRAL
PLATFORMS AND 3D VIRAL ASSAYS
PER TIES UNDER APP HAVING
PANDEMIC OR AT LEAST SIGNIFICANT
OUTBREAK POTENTIAL. SO TO DO
THIS WE FIRST WORKED TO IDENTIFY
WHAT WOULD BE NECESSARY CELL
BASED AND TISSUE BASED MODELS
FOR EACH PRIORITY AREA OF
INTEREST AND THEN TAKE INTO
ACCOUNT FEASIBILITY WE DEVELOP
OR ADOPT HIGH THROUGH PUT
COMPATIBLE ANTI-ADJUVANT MODEL
CELL BASED MODELS THEN USE TO
SCREEN LARGE LIBRARIES TO
IDENTIFY ANTIVIRALS FOR THAT
FAMILY OF INTEREST IF POSSIBLE
WITH GOAL OF WORKING WITH NCATS
BIOCHEMICAL ASSAY TEAMS FOR
VALIDATE VIRAL TARGETS AND
MEDICINAL CHEMIST WHO IDENTIFY
POTENTIAL SMALL MOLECULE FOR
ANTIVIRAL DEVELOPMENT. SO WE ARE
IN PARALLEL DEVELOPING 3D TISSUE
MODELS WORKING WITH MEMBERS OF
THE 3D TISSUE BIOPRINTING LAB
WITH THE MODULAR COMPLEXITY WHEN
POSSIBLE FOR MODELING VIRAL
INFECTION AND VIRAL INDUCED
DISEASE SO THE GOAL IN OUTREACH
MODEL TO STUDY VIRAL INFECTION
AND DISEASE IS DRIVEN BY THE
UNDERSTANDING THAT FOR EACH
VIRUS THE COURSE OF INFECTION
VIRAL DISEASE IMPACT DIFFERENT
DESIRED TREATMENT STRATEGIES SO
FOR EXAMPLE FOR SOME VIRUSES WE
MAY REALLY ONLY NEED ANTIVIRALS
WHEREAS OTHERS WE MAY NEEDS TO
LOOK AT ANTI-MODULATORY DRUGS
FOR MEDIATING RESPONSE SYSTEMIC
DISEASE. SO WE HOPE THESE VIRAL
DISEASE MODELS TISSUE MODELS CAN
ENABLE US TO USE THESE MODELS TO
VALIDATE POTENTIAL THERAPEUTIC
FOR CLINICAL DEVELOPMENT AND
HAVE ON HAND LARGE BODY OF
INFORMATION WHEN MOLECULES OR OR
THERAPEUTICS MAY WORK FOR THE
PREPARATION OF NEXT EMERGING
VIRAL DISEASE FOR RAPID
EVALUATION. SO ALTHOUGH THERE
ARE MANY MODELS THAT WOULD LOVE
TO TALK TO YOU ABOUT TODAY VIRUS
TISSUE PRINTING LAB FOR TIME OF
THIS WORKSHOP I WILL BE FOCUSING
ON THREE EXAMPLES HOW WE ARE
TACKLING 3D MODELS FOR VIRAL
INFECTIONS IN THREE SECTIONS. I
WILL HIGHLIGHT EXPERIENCE
WORKING WITH ALI LUNG MODELS,
FOR SARS COV-2 ANTIVIRAL
VALIDATION AND CHARACTERIZATIONS
OF THE MODEL WE FEEL WE CAN USE
FOR FURTHER THERAPEUTIC
EVALUATION AS REPRESENTATION OF
WHAT KIND OF MODEL WE MAY NEED
FORESTAY DIG DRUGS ANTIVIRALS
AND WHERE WE CAN GO WITH THESE
MODELS WHERE WE THINK WE CAN GO
WITH MODELS IN TERMS OF VIRAL
INFECTION DISEASE AND RELIABLE
READ OUTS. IN A SECOND I WILL
TALK MORPHINETYPIC MODEL SHOWING
EARLY STAGE VIRAL DISEASE
VASCULAR DAMAGE MODEL USING
DENGUE VIRUS IN A MULTI-CHIP
VASCULARIZED TISSUE MODEL AND
THIRD PART TALK ABOUT
REPURPOSING MODELS TO REITERATE
DISCUSSION THESE HUMAN BASE
TISSUE MODELS SHOULD BE
MULTI-PURPOSE AND MULTI-DISEASE
AND SO AS AN EXAMPLE WE CREATED
THE NEURONAL SPHERING MODELS
OVER YEARS FOR PURPOSE OF
MODELING NEUROLOGIC DISEASE SUCH
AS PARKINSON AND ALZHEIMER'S AND
THE IDEA EXPANDING USE OF THIS
MODELS TO VIRAL INFECTION SO
ALSO TOUCH BRIEFLY WHAT WE FEEL
IS NECESSARY FOR THE NEXT STEPS,
MAKE THIS MODEL MORE
REPRESENTATIVE FOR NEUROTROPIC
VIRAL INFECTION WHICH IS TO
FUNCTIONAL BLOOD BRAIN BARRIER
TO THE MODEL. SO START WITH
PANDEMIC A LOT OF US STARTED
WORKING TOGETHER TO GENERATE NEW
OR REPURPOSE EXISTING MODELS FOR
SARS COV-2 INFECTION. THE
STRATEGY WAS TO FOCUS ON THE
RESPIRATORY TRACT WHICH IS
IMPORTANT TO COVID WITH FOCUS ON
DIFFERENT AREAS OF DIFFERENT
CELL COMPOSITIONS INCLUDING THE
AIRWAY BRONCHIAL REGION AND
ALVEOLI SO WE WORKED IN VARIOUS
MODELS REPRESENTING DIFFERENT
LEVELS OF PHYSIOLOGICAL
COMPLEXITY RANGING FROM THE
SIMPLEST LUNG TRANSWELL ASH
LIQUID INTERFACE MODEL TO
CO-CULTURES OF PULMONARY CELL
TYPES TO BIOPRINTING OF VASCULAR
BED ALONG EPITHELIA TO
REPURPOSING THE 64 CHIP ORGANO
GRAPH PLATE TUMOR FOR
VASCULARIZED SPHEROIDS AND
AIRWAY OR ALVEOLAR EPITHELIAL
CELLS IN TOP OF PERFUSABLE
PULMONARY VASCULAR BED. WE HAVE
TO THINK ABOUT WHAT THE NEXT
STEPS WE WOULD TAKE WHAT ARE
ADDITIONAL CELL TYPES NECESSARY
TO ADD THERE IS TALK ABOUT
IMMUNE CELLS IN THIS WORKSHOP
AND WHAT READ OUTS WE NEED TO
EACH MODEL FOR REPRODUCIBLE
SYSTEMS FOR THESE TO REALLY BE
USED IN TRUE DRUG DISCOVERY
FASHION. IN ADDITION TO
IMPORTANCE OF CREATING UNBIASED
PARAMETERS USED BY ANY GROUP FOR
UNBIAS QCs THE MODELS
HOPEFULLY EVALUATING ANY
INFECTION OR DRUG EFFICACY DATE
USE IN HOUSE MODELS FOR VIRUS,
SO WE MAKE OUR OWN IN HOUSE
SMALL AIRWAY AND ALVEOLAR 96
TRANSWELL MODEL AND PURCHASED
MATTEK AT ALVEOLAR MODELS SO
THESE CREATED BY PRIMARY LUNG
EPITHELIAL CELLS WITH OR WITHOUT
FIBROBLASTS ON TO TRANSWELL
MEMBRANE FORM EPITHELIAL LAYER
ON THE TOP AND MOVING CULTURE TO
AIR TO ALLOW THEMSELVES TO
MATURE, YOU SEE EMERGENCE OF
SEVERAL POPULATIONS IN THESE
MODELS IN THE AIRWAY YOU WILL
SEE POPULATION OF SILLIATED
CELLS, GOBLET CELLS AND
POPULATION CELLS ALVEOLAR TYPE
ONE AND TWO AND INTERMEDIATE
CELLS SO WE USE FOR VARIETY OF
INPEEKSES TO FOCUS ON SARS
COV-2, WHICH CASE FOR DRUG
SCREENING WE ADD MOLECULES TO
BASAL CHAMBER TO APICAL SIDE AND
MEASURE AMOUNT OF VIRAL
INFECTION ON THE TISSUE BY
DIRECT ANTIGEN STAINING FOR
BROWN CELLULAR MARKERS LOOK AT
TISSUE STRUCTURE MEASURE
SECRETED VIRUS, SECRETED
CYTOKINE CHEMOKINES AND DOING
SINGLE CELL RNA SEQ WORK IN
HEALTHY VERSUS INFECTED TISSUE
TO CHARACTERIZE DISEASE VERSUS
HEALTHY STATE AFTER SARS COV-2
INFECTION. SO YOU CAN SEE HERE
THAT THESE AIRWAY ALVEOLAR
TISSUES CELL MENTION WED ARE
USING THE TECH TISSUES IN THESE
NEXT FEW SLIDES SUPPORT
PRODUCTIVE INFECTION COV 2
MEASURING THE AMOUNT OF RELEASE
INFECTIOUS VIRAL PARTICLES AND
THAT VARIANCE DO HAVE DIFFERENT
INFECTIVITY OF THIS TISSUE SO WE
CAN LOOK WHETHER THERE ARE ANY
CELLULAR TROPISM COV 2 TISSUE
TYPES BY IMMUNOSTAINING AND WHAT
WE SEE IS PRIMARY TARGET IN
THESE BRONCHIAL TISSUES
SILLIATED CELLS, WHEN WE LOOK AT
VARIANCE OF ALPHA BETA GAMMA AND
DELTA SARS COV-2 ONE OF OUR
GOALS IS TO CHARACTERIZE ANY
TISSUES DEVELOPED AS MUCH AS
POSSIBLE BECAUSE PART OF OUR
MISSION IS ENABLE OTHER GROUPS
TO BE ABLE THE USE THESE
TECHNOLOGIES WITHOUT HAVING TO
REINVEST RESOURCES TO
RECHARACTERIZE THE SAME MODEL SO
BECAUSE OF THIS WE STARTED USING
SINGLE CELL RNA SEQ ON BOTH
HEALTHY AND DISEASE TISSUE AND
SHOWN HERE IS AN EXAMPLE LOOK AT
WHAT TISSUE VIRAL SPECIFIC
TRANSCRIPTOMIC PROFILES ARE IN
THAT WE OBSERVE IN AIRWAY VERSUS
ALVEOLAR ALI TISSUE MODEL IN
SARS COV-2 VERSUS HEALTHY
INFLUENZA INFECTION. BUT THE
PURPOSE OF WHETHER THIS CAN BE
LINKED TO DISEASE OBSERVED IN
VIVO WITH GOAL BEING WHEN WE
TEST THERAPEUTIC AGENT AGAINST
SARS COV-2 INFECTION DO WE GET
RETURN TO HEALTHY STATE VERSUS
DISEASE STATE. SO THIS IS EARLY
WORK WE HAVE DONE IN STARTING TO
CHARACTERIZE DISEASE STATES IN
CONTEXT OF INFECTION AND I WON'T
GO INTO MUCH DETAIL BECAUSE OF
TIME BUT WHAT YOU CAN SEE HERE
IS THE TISSUE RESPONSE TO VIRAL
INFECTION IS DEPENDENT ON THE
VIRUS AND ON THE TISSUE TYPE.
AND AS AN EXAMPLE, THIS IS THE
EARLY WASHINGTON SARS COV-2
VARIANT YOU CAN SEE TISSUE
RESPONSE TO SARS COV-2 MORE
SUBTLE COMPARED TO INFLUENZA BUT
STILL GETTING CHANGES TO PROFILE
SO WE CAN START TO IDENTIFY
DIFFERENT PROFILES OBSERVED
BETWEEN DIFFERENT VIRUSES AND
THE TISSUE BETWEEN SAME VIRUS
AND DIFFERENT TISSUES AND WHAT
WE HOPE TO BE ABLE TO CREATE IS
A PANEL OF BIOMARKERS THAT WE
CAN USE TO EVALUATE SEVERE
DISEASE IN DIFFERENT TISSUES OF
THE RESPIRATORY TRACK. SO THIS
IS AN EXAMPLE, INTERESTED IN
INFECTION INFLAMMATORY RESPONSE
AIRWAY VERSUS ALVEOLAR ISSUES
THAT AFFECTS SEVERITY OF DISEASE
AND INFECTED INDIVIDUAL SO WE
LOOK AT PANEL LINKED TO DISEASE
IN COVID PATIENTS WE SEE HIGH
INDUCTION OF IP 10 PLURIPOTENT
CHEMOKINE. ALI TISSUE IN
PARTICULAR WE WERE OBSERVING A
HIGH INFECTION IN BOTH ALVEOLAR
EPITHELIAL CELLS AND WITH
CONTRIBUTION ENDOTHELIAL CELLS
AND FIBROBLASTS AS WELL. SO WE
WANTED TO USE THE MODELS DURING
EARLY DAYS PANDEMIC. WE DIDN'T
HAVE ACCESS TO BSL 3 UNLIKE NOW
SO WE ARE LUCKY TO BE ABLE TO
COLLABORATE WITH DR. SARA CHERRY
AT UPENN WE HEARD SPEAK BEFORE
LUNCH, DR. CHERRY ARE WAS
WORKING WITH HUMAN CELL BASED
SCREENS AND TISSUE MOLES TO
UNCOVER DIFFERENCES IN ANTIVIRAL
ACTIVITY IN DIFFERENT HUMAN AND
NON-HUMAN CELL TYPES SO WE
STARTED WORKING WITH DR. CHERRY
PAIRING TISSUES MAYBE PREDICTIVE
AND WHETHER THIS HELPS NARROW
DOWN COMPOUNDS FOR FURTHER
EVALUATION IN ANIMAL MODELS
PRE-CLINICAL DEVELOPMENT. SO
THIS IS SHOWING SOME OF THE
DRUGS AND READ OUTS WE USE
RANGING FROM DIRECT VIRAL
ANTIGEN STAINS, QR PCR
PRODUCTION AND WHAT YOU CAN SEE
HERE IN THE BOTTOM RIGHT CORNER,
WE HAVE THE HYDROXYCHLOROQUINE,
EXPLORED EARLY ON IN THE
PANDEMIC DOESN'T WORK IN MODELS
DISCUSSED TODAY AND YESTERDAY
AND SUGGESTED BEFORE IT IS
POSSIBLE IF MODELS WERE
WIDESPREAD AND WIDESPREAD USE
EARLY ON DURING THE PANDEMIC WE
WOULD HAVE RECOGNIZED ANTS VIRAL
ACTIVITY MUCH EARLIER SO A KEY
TAKE AWAY FROM OUR EXPERIENCE IS
THAT WE NEED TO FULLY
CHARACTERIZE THESE MODELS EARLY
BEFORE THE NEXT VIRAL OUTBREAK
WITH BOTH DIRECT VIRAL INFECTION
READ OUT AND DISEASE
INFLAMMATORY READ OUTS, AND HAVE
THIS ACCESSIBLE FOR VIRAL FAMILY
AND TISSUES OF INTEREST IN THIS
CASE RESPIRATORY TRACK SO THE
NEXT VIRAL AGENT ARISES WE HAVE
THESE TISSUES AND TECHNIQUES IN
OUR TOOLBOX READY FOR
THERAPEUTICS EVALUATION SO THAT
WE CAN HAVE A MORE REAL TIME
RAPID IMPACT VIRAL THERAPEUTICS
BEING CONSIDERED FOR DEPLOYMENT
SO FOR PART 2 OF MY TALK I WOULD
LIKE TO TALK ABOUT VIRAL INDUCED
DISEASE MODELING THIS IS AN
EXCEPTION OF THE FIRST PART
BECAUSE WE ARE INTERESTED IN IN
DIRECT INFECTION. SO WE ARE ALSO
LOOKING AT MORE SEVERE DISEASE
READ OUTS. DENGUE VIRUS IS A
POSITIVE SENSE RNA VIRUS, POSES
A HUGE GLOBAL BURDEN EACH YEAR
WITH AROUND 400 MILLION CASES
ESTIMATED PER THE WORLD HEALTH
ORGANIZATION PER YEAR WORLDWIDE
WITH SEVERE DENGUE AFFECTING
SUBSET OF INDIVIDUALS THAT
RANGES DEPENDING ON AREA. SO AS
MENTIONED EARLIER OUR LAB WORKED
ON DEVELOPING BOTH AIRWAY
ALVEOLAR AND CHIP MODELS ORGANO
GRAPH PLATE GRAPH WHICH IS A 64
CHIP PROFUSE TISSUE MODEL THAT
USES HERE ON THE RIGHT TO INDUCE
FLOW IN THE MODEL. SO BECAUSE WE
KNOW THAT DENGUE VIRUS CAUSES
VASCULAR DAMAGE IN CASES OF
SEVERE DENGUE WE SOUGHT TO
MODIFY THE MODEL TO CREATE A
SIMPLER MODEL TO START FOR
STUDYING DENGUE INDUCED DAMAGE.
SO IMPORTANTLY, REASONS WE
SOUGHT TO USE THIS MODEL IS THAT
ONE, IT IS 64 CHIPS PER PLATE SO
WE CAN USE IT AND IT USES
IRONINGER SO WE CAN USE BSL 2
AND 3 CONTEXT AND IT IS
PROFUSABLE WHICH WILL ALLOW US
TO LOOK AT VIRAL EFFECTS ON
VESSEL WEAKNESS AND SHOWN HERE
ARE EXAMPLE PROFUSIONS AND GROUP
SMALL AIRWAY CHIP MODEL THAT
(INDISCERNIBLE) MADE IN THE LAB
WE ARE SHOWING PROFUSION WITH 70
KD FLUORESCENT STRAND. WE ARE
OPTIMIZING THE TISSUE TO
MAINTAIN LARGE ENOUGH
CONNECTIONS BETWEEN THE LARGE
VESSELS AND MICROVASCULAR
NETWORK WHEN INCORPORATING
DIFFERENT EPITHELIAL LAYERS
BECAUSE THAT DOES IMPACT THE
VASCULATURE UNDERNEATH. SO WE
ARE DOING THIS AS WE EVALUATE
DIFFERENT CELL TYPES PERMISSIVE
TO DENGUE INFECTION. SO FOR OUR
FIRST PILOT STUDIES WE ARE USING
GFP EXPRESSING HUMAN PULMONARY
ENDOTHELIAL CELLS, WITH SEE
ENDOTHELIAL CELLS AND PAIR SITES
IN LOWER DENSITY LOWER CHAMBER
HYDRA GEL AND ALLOW THEM TO FORM
THIS NETWORK OVER TIME. BECAUSE
THEY ARE GFP WE CAN QC TISSUE
PLATE OVER TIME AND EXCLUDE
MONOLAYERS OR USE THE -- WE CAN
ALSO USE THE INFORMATION TO
INFORM US OF CHANGES IN THE
TISSUE RELATED TO INFECTION OR
SIMPLY RELATED TO AGING OF
TISSUES. SO SHOWN HERE ARE
REPRESENTATIVE IMAGES OF
MONITORING THE MICROVASCULAR
NETWORK DEVELOPMENT OVER TIME
FROM SEEDING UP UNTIL
MATURATION. BY USING THE
EPITHELIAL CELLS AND MON FORKING
THE SAME LEVEL FOR VIRAL INSULTS
IN THE BOTTOM WE ARE ABLE TO
OBSERVE MORPHOLOGICAL CHANGES IN
TISSUE AFTER RECOMBINANT DENGUE
TO THE CHANNEL IN VIRUS
EPITHELIAL LAYER QUANTIFY SHOWN
IN THE GRAPH IN THE LEFT AND WE
ARE CURRENTLY EXPLORING
ADDITIONAL PARAMETERS SUCH AS
VESSEL BRANCHING WEAKNESS AS
WELL AS ENDOTHELIAL DYSFUNCTION
SUCH AS SILL YAK ACID SECRETION.
I WILL NOTE HERE WE ARE USING A
DENGUE PERMISSIVE EPITHELIAL
LAYER OF EOSINOPHILS WHICH IS
IDEA WE USE THIS AS WE CREATE A
MODEL AND THEN SWITCH TO A
PRIMARY LINE. SO AS I MENTION
WE ARE ALSO ACTIVELY SEEKING TO
REPURPOSE EXISTING MODELS FOR
THE PURPOSE OF VIRAL INFECTION
AND DISEASE MODELING, AND AS
DIVERSE RANGE OF TISSUES AS
NECESSARY FOR VIRUSES
PRIORITIZED UNDER ATP. SO WE
RECENTLY PUBLISHED A PRE-PRINT
DESCRIBING BRAIN REGION SPECIFIC
NEUROSPHERING MODEL WE MAKE IN
OUR LAB FOR DIFFERENT
NEUROLOGICAL DISEASE INCLUDING
ALZHEIMER'S DISEASE AND
PARKINSON. WE MAKE THESE
SPHEROIDS IN 3D PLACE USING IPSC
DIFFERENTIATED MATURED NEURONS
AND ASTROCYTES AND WE CAN ALTER
THE GLUTAMATERGIC GABAERGIC
NEURONS TO RATIOS REPORTED TO BE
PRESENT IN DIFFERENT BRAIN
REGIONS AND IMPORTANTLY WE CAN
SEE FUNCTIONAL SYNAPSES FORMING
LOOKS LIKE MY VIDEO IS NOT
PLAYING. THEN WE ALSO CAN SEE --
WE CAN ALSO USE KALIUMS IMAGING
TO SEE CORRESPONDING DIFFERENT
ACTIVITY PROFILES DEPENDING ON
RATIO OF NEURAL SUBTYPE
INCORPORATION. AN ADVANTAGE OF
USING CALCIUM ACTIVITY AS READ
OUT FOR THIS MODEL WE CAN LOOK
AT THE SAME SPHEREROID OVER TIME
BY TRANSDUCING EARLY ON DURING
MATURATION USING AN AAV VIRUS
ENCODING FOR GENETICALLY ENCODED
CALCIUM INDICATOR AND THEN
MEASURE AFFECTS OF DRUGS OR
INSULTS ON NEURONAL FIRING AND
THE SPHEROID USING A WHOLE PLATE
FLUORESCENCE IMAGING PLATE
READER OR LOOK MORE CLOSELY
USING AN AUTOMATED CONFOCAL
MICROSCOPE TO MEASURE INDIVIDUAL
AGENTS OF INTEREST AND NOT
CHANGES IN SYNCHRONIZATION OVER
TIME SO ON THE LEFT IS CONFOCAL
IMAGING AND ON THE RIGHT IS THE
FLUORESCENT IMAGING PLATE READER
THAT WE USE. SO PROOF OF
CONCEPT WE EXPOSE THESE
SPHEROIDS TO SARS COV-2, WITHIN
ABLE TO DETECT ROBUST INFECTION
OF THE NEURONAL SPHEROID SHOWN
BY GREEN STAINING SIGNAL,
STAINING FOR THE SARS COV-2 KNEW
CLEAR CAPSID PROTEIN, BUT WHILE
SARS COV-2 IS LIKELY NOT TROPIC
VIRUS WE ARE GOING TO BE
EXPANDING THIS SO RECENTLY POST
DOC WITH EXPERIENCE IN
NEUROTROPIC VIRUSES SHE WILL BE
EXPANDING THIS MODEL TO A
VARIETY OF THE SL 2 LEVEL
NEUROTROPIC VIRUSES TO START
INVESTIGATING SHORT TERM VERSUS
LONG TERM CHANGES TO NEURON
SPHEREROID ACTIVITY AS WELL AS
SYNCHRONIZATION AS ONE OF THE
LIMITATIONS OF THE SYSTEM IS
HAVING ACCESS TO EQUIPMENT IN
APPROPRIATE BIOSAFETY LEVEL
SPACE SO THIS BRINGS UP ANOTHER
CRITICAL QUESTION ABOUT THIS
MODEL WHICH IS WHETHER WE CAN
USE THIS TO EVALUATE
NEUROTROPISM EMERGING VIRUSES,
WHICH I THINK TO DO THIS IN A
REPRESENTATIVE WAY WE MIGHT
FIRST VIRUS CAN INFECT NEURONAL
CELLS BUT THEN CHECK IF WE CAN
INCORPORATE BLOOD BRAIN BARRIER
INTO THIS MELD WE CAN EVALUATE
PAST BLOOD BRAIN BARRIER BBB
ACTS AS A BARRIER TO VIRAL
INFECTION IN THE BRAIN. TO WRAP
UP THIS TALK OUR ULTIMATE GOAL
TO FULLY CHARACTERIZE ISSUE IN
CONTEXT OF OUR OWN RANGE OF
INFECTION AS PRIORITIZE UNDER
ATP OF INTEREST. RANGING FROM
RESPIRATORY TRACK, BRAIN GUT
KIDNEY CARDIAC, WE STILL HAVE A
LONG WAY THE GO AND FOR THAT WE
HOPE TO COLLABORATE WITH EXPERTS
IN THE FIELD TO HELP SUPPORT
THESE CRITICAL EFFORTS AND
UTILIZING THE 3D TISSUE MODELS
FOR ANTIVIRAL RESEARCH FOR THE
BROADER SCIENTIFIC COMMUNITY.
WITH THAT, I WOULD LIKE TO
HIGHLIGHT THE LARGE NUMBER OF
INDIVIDUALS INVOLVED IN THIS
WORK WITH INDIVIDUALS
SPECIFICALLY INVOLVED HERE
HIGHLIGHTED. AND LIKE TO SAY
THANK YOU TO THE WHOLE 3D LAB
AND ATP TEAM AND TO OUR
COLLABORATORS FOR ENABLING THIS
WORK. THANK YOU FOR YOUR TIME
AND LISTENING.
>> THANK YOU SO MUCH FOR THE
EXCELLENT TALK. LOOK FORWARD TO
COMING OUT AS PART OF THE
DISCUSSION IN A BIT. NOW I WILL
TRANSITION OVER TO OUR FINAL
PRESENTATION OF THE SESSION 2
AND INTRODUCE DR. CALVIN KUO.
DR. KUO IS THE MAUREEN LILLS
DAMBROSIO PROFESSOR OF MEDICINE
STANFORD UNIVERSITY SCHOOL OF
MEDICINE. HIS RESEARCH
INTERESTS INCLUDE MODELING OF
IMMUNITY AND PATHOGENS AND
CANCER IN 3D ORGANOID CULTURES
BIOLOGY OF INTESTINAL AND LUNG
STEM CELL POPULATIONS, AND
MOLECULAR REGULATION OF
ANGIOGENESIS AND THE BLOOD BRAIN
BARRIER. HE IS ELECTED MEMBER OF
ASSOCIATION OF AMERICAN
PHYSICIAN, AMERICAN SOCIETY FOR
CLINICAL INVESTIGATIONS AND THE
AAASS. TODAY HE WILL BE TALKING
TO US THE TITLE OF HIS TALK IS
USING ORGANOID MODELS OF TISSUE
RESIDENT IMMUNITY TO STUDY
INFECTIOUS DISEASESS. DR. KUO.
OVER TO YOU.
>> HOPEFULLY YOU CAN HEAR ME.
LET ME TRY THE SHARE SCREEN
HERE. CAN YOU SEE THIS?
>> CURRENTLY I SEE YOUR VIDEO
NOW. NOT YOUR SLIDES.
>> VIDEO NOT SLIDES.
>> YEAH. IS IT NOT JUST ME?
>> YEAH WE CAN'T SEE THE SLIDES
ABIGAIL, DO YOU WANT TO JUMP IN
AND HELP?
>> GIVE IT A GO.
>> HANG ON, I KNOW WHAT TO DO. I
THINK. THAT SHOULD PROBABLY
WORK. GREAT. SO THANKS FOR THIS
KIND INVITATION AND I WOULD LIKE
TO TELL YOU ABOUT OUR RECENT
WORK IN ORGANOID MODELING OF
INFECTIOUS DISEASE AS WE KNOW
THERE IS DIFFERENT TYPES OF
ORGANOIDS, THERE IS WE
SPECIALIZE IN ADULT HUMAN
ORGANOID TYPE, NOT IPSC DERIVE
AND WE HAVE BEEN WORKING BOTH
ORGANOIDS THAT HAVE EPITHELIUM
ONLY AND THEN EPITHELIAL PLUS
STROMA, I WILL MENTION BOTH OF
THOSE TO YOU GUYS. STARTING WITH
EPITHELIAL ONLY ORGANOIDS, AS
YOU HEARD FROM EARLIER TALKS
THERE HAS BEEN A TREMENDOUS
EXPLOSION IN THE ABILITY TO
CULTURE ADULT HUMAN TISSUES. AS
ORGANOIDS. THIS IS ONLY HALF OF
WHAT OUR LAB DOES SO IT IS
CLEARLY A ROBUST METHODOLOGY AND
WE PERTINENT TO THIS CONFERENCE
WE HAVE BEEN INTERESTED IN ADULT
DISTAL LUNG ORGANOIDS AT THE
TIME WE INITIATED THE PROJECT IT
WAS VERY DIFFICULT PREVIOUSLY TO
GROW HUMAN ALVEOLI IN A FEEDER
FREE CHEMICALLY DEFINED CLONAL
FASHION. SO WE STARTED WORKING
ON THIS SIX OR SEVEN YEARS AGO.
CAME UP WITH METHODOLOGIES TO
GROW BOTH THE ALVEOLAR
COMPONENTS AS WELL AS MORE BASAL
AIRWAY COMPONENTS AS CLONAL
ORGANOIDS AND YOU CAN SEE THE
SOLID MORE BASAL AIRWAY
ORGANOIDS AND THE CYSTIC
ALVEOLAR ORGANOIDS. THEN WORKING
WITH MANUEL LED BY M.D. Ph.D.
STUDENT, WE ALSO HAVE BEEN USING
THIS ROBUST SUSPENSION CULTURE
THAT MANUEL'S GROUP DEVISED THAT
CAN TURN ORGANOIDS INSIDE OUT.
BASICALLY TAKE OUR ORGANOIDS
THAT HAVE HAD A LOT OF THE
DIFFERENTIATED CELLS ON THE
INTERIOR, WHY YOU ARE NOT SEEING
THE GREEN AND WHITE SIGNAL IN
SURFACE VIEW LEFT-HAND SIDE, YOU
CAN PUT THEM INTO SUSPENSION
CULTURE AND EVERT THE ORGANOIDS
LIKE A SOCK THAT IS THE WRONG
WAY IN THE LAUNDRY YOU GET
SILLIATED CELLS IN WHITE AND
GREEN CELLS INITIALLY INCITE --
INSIDE NOW ON THE OUTSIDE. THAT
WAS IMPORTANT BECAUSE IN OUR
ORGANOID IT IS ACE 2 RECEPTOR
WAS INTERIOR AND THESE ARE
APICAL IN ORGANOIDS WHEREAS IN
SUSPENSION CULTURE ACE 2 IN
GREEN IS ON THE OUTSIDE AND WE
WANTED TO USE THAT FOR INFECTION
STUDIES YOU CAN SEE THE
SILLIATED CELLS HERE IN THE
UNEVERTED SITUATION VERSUS NOW
AFTER E VERSION AND YOU CAN SEE
THTHE CILIARE ARE ON THE OUTSID.
THIS ALLOWED US TO BE ABLE TO DO
THESE INFECTIOUS STUDIES BY
ADDING SARS COV-2 TO THE
EXTERIOR OF THE NOW FLIPPED
ORGANOIDS, WONDERFUL
COLLABORATION WITH CATHERINE
BLISS, AND SO WE HAVE BEEN ABLE
TO DEMONSTRATE THAT NUCLEAR
CAPSID STAIN AND PLAQUE ASSAYS
AND ENVIRONMENT GENOMES DETECTED
WE HAVE BEEN ABLE TO SHOW THE
CLUB CELLS ARE A NOVEL CELL TYPE
INFECT AND OBVIOUSLY WE ARE
GETTING VERY ROBUST INFECTION OF
ALVEOLAR TYPE 2 ORGANOIDS. WE --
I THROUGH THIS IN BECAUSE WE
HAVE BEEN PLAYING AROUND WITH
MULTI-WELL SCREENING OF THESE
TYPES OF ORGANOIDS. MORE FOR
INFLUENZA ANTIVIRAL COMPOUNDS
BUT CERTAINLY ADAPTABLE TO OTHER
APPLICATIONS AS WELL I WOULD
SAY. AND WE CAN PLATE THESE IN
TO 96 WELL DISH, GET INFECTION,
THIS IS INFLUENZA H 1N 1 GFP
STRAIN SO YOU CAN SEE THE GFP
THEN WE CAN APPLY VARIOUS
ANTIVIRAL COMPOUNDS IN THIS
MULTI-WELL FORMAT AND GET
INHIBITION. SO VERY POSSIBLE I
WOULD SAY TO TAKE THIS FIRST
GENERATION TYPE OF ORGANOID AND
PERFORM DRUG SCREENING. I WILL
SWITCH TO WORK ON EPITHELIAL
PLUS STROMAL ORGANOIDS. THESE
ARE MORE COMPLEX AND HAVE I
WOULD SAY VERY HOLISTIC
REPRESENTATION OF THE TISSUE
MICROENVIRONMENT. WE HAVE BEEN
USING AIR INTERFACE YOU HAVE
HEARD ABOUT. IN MULTIPLE TALKS
BUT WE ARE DOING 3D AIR LIQUID
INTERFACE SO OUR INITIAL EFFORTS
STARTED WITH INTESTINE BACK IN
2009. I WILL SAY WE HAVE BEEN
MORE RECENT YEARS USING THIS FOR
CANCER CULTURES. THIS IS JUST A
WHOLE VARIETY OF DIFFERENT TYPES
OF TUMORS FROM ANATOMIC SITES WE
CAN GROW USING THIS TECHNIQUE.
THE IMPORTANT THING IS THIS
METHODOLOGY, WE START OFF WITH
TISSUE FRAGMENTS NOT DISSOCIATED
CELLS SO WE PRESERVE
MICROENVIRONMENT WITHIN THE
FRAGMENTS WE ARE CULTURING. SO
IN YELLOW YOU CAN SEE THE CANCER
FIBROBLAST STROMA EMBEDDED
WITHIN THE TUMOR ORGANOID WITH
COLON PATIENT DERIVED ORGANOID.
IMPORTANTLY THIS TYPE OF
TECHNIQUE PRESERVES RESIDENT
IMMUNE CELLS WITHOUT ANY
ARTIFICIAL RECONSTITUTION. THIS
IS A RENAL CELL CARCINOMA
ORGANOID WITH TUMOR CELLS IN
PINK AND TUMOR INFILTRATING
LYMPHOCYTES CD 3 MEMBRANE
STAINENING YELLOW. WE DIDN'T
RECONSTITUTE FROM PERIPHERAL
BLOOD THIS IS TISSUE RESIDENT OR
IE TUMOR RESIDENT FOR
LYMPHOCYTES ON THE LEFT AND
THESE ORANGE MACROPHAGES ON THE
RIGHT. SO WE CAN PRESERVE IN
SITU THE TISSUE RESIDENT
POPULATIONS SO THIS IS ALLOWS US
TO DO THINGS LIKE CHECK POINT
INHIBITORS AND TUMOR ORGANOIDS
AND GET THE TUMOR INFILTRATING
LYMPHOCYTES TO ATTACK TUMOR
CELLS. BUT I WANT TO SWITCH FROM
CANCER TO INFECTIOUS DISEASE.
WE CAN USE THE SAME TYPES OF
TECHNIQUES THAT HAVE ORGANOIDS
AND INTRINSIC IMMUNE CELLS
INFILTRATING IMMUNE CELLS AND
THEN ADD PATHOGENS. WE HAVE OF
COURSE POSTER CHILD IS SARS
COV-2 WE WANTED TO ESTABLISH
THESE TYPES OF LUNG ORGANOIDS
THAT HAVE PRIMARY EPITHELIAL
INFECTION BUT LOOK AT SECONDARY
IMMUNE ACTIVATION PARTICULARLY
ADAPTIVE RESPONSE OF T AND B
CELLS WITHIN THESE ORGANOIDS.
SO OUR M.D. Ph.D. STUDENT HAS
BEEN LEADING THE CHARGE ON THIS
AND THIS HAS BEEN A VERY ROBUST
TECHNIQUE WITH THREE DIMENSIONAL
ALI NOT TWO, AND YOU CAN SEE THE
CONTINUED GROWTH OVER 180 DAYS.
AND BY HNE THERE'S ALL SORTS OF
ALVEOLAR AIR SACKS AND
BRONCHIOLAR STRUCTURE THAT ARE
APPARENT BY STAINING THERE IS
CLUB CELLS SILLIATED CELLS,
BASAL CELLS, WE HAVE BOTH
ALVEOLAR TYPE 1 AND 2 CELLS AND
DISTINCT DOMAINS. AND THE
ALVEOLAR TYPE 1 IN GREEN YOU CAN
SEE FORMING THESE ALVEOLAR HONEY
COMB STRUCTURE WITH SOME DEGREE
OF INTEGRATION OF ALVEOLAR TYPE
2 CELLS IN THEM BUT WE FIND AT 2
CELLS DO GROW OUTSIDE OF THE
ALVEOLI AS WELL. I WILL MS.
MENTION WE HAVE EXTENSIVE
ENDOTHELIAL NETWORKS WITHIN THE
ORGANOIDS. THIS IS CD 31
STAINING YOU CAN SEE THAT THERE
IS VERY ARBORIZING TYPE OF
VASCULAR NETWORKS. NO PROFUSION
PER SE BUT IT WILL BE
INTERESTING TO BUILD IN
SUSTAINED PROLIFERATION WE CAN
SEE THE SF SUR FACT TIN PROTEIN
C DOMAIN IN WHITE HAS KI 67
POSITIVE CELLS WITHIN. P P AND
THEN WE ALSO HAVE MESENCHYMAL
STROMA. THIS IS STAINING IN RED
AND EPITHELIAL CELLS IN GREEN
BUT MORE IMPORTANTLY WE HAVE
IMMUNE CELLS, THIS IS CD 45
STAINING HEMATOPOIETIC MARKER.
WE ARE NOT RECONSTITUTING FROM
PERIPHERAL BLOOD THESE ARE
TISSUE RESIDENCE IMMUNE CELLS WE
PRESERVE BAKED INTO THE
ORGANOID. THEN WE CAN USE OUR
SUSPENSION CULTURE METHOD TO
EVERT VERY COMPLEX ORGANOID THAT
IS PROBABLY 10 TO 50 TIMES
LARGER THAN ORIGINAL EPITHELIAL
ONLY LUNG ORGANOIDS. WHEN WE DO
THAT IMMUNE CELLS CD 45 ARE
STILL PRESERVED WITHIN THE
ORGANOIDS. AND THE ACE 2 IN RED
WHICH WAS ORIGINALLY GRITTED
DIFFUSELY IS NOW EXTENDED ON THE
OUTSIDE IN A MORE ACCESSIBLE WAY
FOR INFECTION STUDIES. SO BY
SINGLE RNA SEQUENCING OF LUNG
ORGANOIDS WE CAN FIND THAT ANY
EPITHELIAL COMPARTMENT THERE IS
TREMENDOUS DIVERSITY WITHIN
THESE ORGANOIDS CERTAINLY
ALVEOLAR TYPE 1 AND 2 CELLS,
PROLIFERATIVE ALVEOLAR TYPE 2
CELLS, BASAL CELLS,
PROLIFERATIVE BASAL CELLS.
FIBROBLASTS, ENDOTHELIAL CELLS,
BUT THEN ON THE IMMUNE SIDE OF
THE ORGANOIDS THERE IS A VERY
TREMENDOUS COMPLEXITY AS WELL
THAT EXTENDS NOT ONLY TO CD4 AND
CD8 T-CELLS BUT TREGS, THERE IS
MYELOID CELLS, MACROPHAGES,
THERE'S B CELLS AND PLASMA B
CELLS WE CAN ALSO DETECT NK
CELLS AND THERE IS ALSO TISSUE
RESIDENCY MARKERS WE CAN DETECT
AS WELL ON THE T-CELLS. SO VERY,
VERY HOLISTIC REPRESENTATION,
TREMENDOUS CELLULAR DIVERSITY
WITHIN THESE ADULT LUNG
ORGANOIDS. NO RECONSTITUTION.
THE TCR REPERTOIRE OF THE
ORIGINAL TISSUE MADE IN THE
ORGANOID INDUCE SINGLE CELL
SEQUENCING AND LOOK AT TCR ALPHA
BETA SEQUENCINGS IN THE CDR 3
REGENERAL, THE SAME -- REGION.
THE SAME CLONE TYPES EXPANDED IN
TISSUE ARE ALSO CLONALLY
EXPANDED IN THE ORGANOID. AND
THIS IS JUST A CORRELATION PLOT
BETWEEN THE TCR REPRESENTATION
BETWEEN ORGANOID AND TISSUE AND
WE SEE A LOT OF CCR 3 SEQUENCES
THAT MATCH PUBLIC DATABASE AND
RIGHT NOW RUNNING SAME SEQUENCE
ANALYSIS FOR SARS COV-2. SO HERE
BSL 3 AND THIS HAS BEEN A VERY
LARGE UNDERTAKING WITH FOLKS IN
MY LAB AND CATHERINE'S LAB,
REALLY DOING A LOT OF THESE
STUDIES IN BSL 3, AND INDEED WE
CAN CERTAINLY INFECT THESE VERY
COMPLEX LEVEL ORGANOIDS WITH
SARS COV-2 AND HERE THIS IS A
HIGH LEVEL OVERVIEW YOU CAN SEE
THE TRINITY OF THE SARS COV-2
INFECTION IN GREEN WITH NUCLEIC
CAPSID STAINING BUT THEN YOU
HAVE THE RED E CAD HEREIN LUNG
EPITHELIUM AND THE CD 45 LUNG
INTRINSIC HEMATOPOIETIC CELLS
PRESENT SO WE HAVE ALL THREE
COMPONENTS ALL TOGETHER. SO IF
THE EXTENT OF INFECTION THAT WE
SEE IN THE LUNG ALI ORGANOIDS IS
MUCH MORE EXTENSIVE THAN WHAT WE
WERE SEEING IN OUR LUNG
EPITHELIAL ONLY ORGANOIDS. WE
CAN GET EXTREMELY SENSITIVE
INFECTION AS I SHOW YOU IN
SUBSEQUENT SLIDES. THEN ALSO
THIS SLIDE SHOWS THEE
DIFFERENCE BETWEEN THE SECOND
GENERATION ORGANOID ON THE LEFT
AND FIRST GENERATION ORGANOID ON
THE RIGHT. SO WE CAN DO VIRAL
STUDIES, THIS IS AN ANTIVIRAL
SCREENING CONFERENCE SO I
CONCLUDED LOTS OF DATA WITH
REMDESIVIR. THIS IS INDICATING
THAT WE CAN INDEED GET SARS
COV-2 REPLICATION WITH PLAQUE
FORMATION BUT THAT REMDESIVIR
RDV CAN STRONGLY INHIBIT THAT IN
PLAQUE ASSAYS. ALSO IF WE LOOK
AT CYTOKINE PRODUCTION BY QRT
PCR YOU CAN SEE THAT THE DARK
ORANGE OR BROWNISH BARS THESE
ARE SARS COV-2 INDUCED CYTOKINE
mRNA BUT THEN WITH RECOMMEND
DECEMBER VEER, THIS IS THE LIGHT
-- REMDESIVIR YOU CAN SEE
SIGNIFICANT INHIBITION OF
IMMUNOSTAINING LOTS OF
INTERFERONS TNF ALPHA, IL 6, ET
CETERA. WE CAN DO SAME WITH
LUMENEX. WE CAN TAKE CULTURE
SUPERNATANT THE ORGANOIDS AND
YOU CAN SEE THE STRONG
STIMULATION FOR INSTANCE WITH IL
6 FOR INSTANCE WITH SARS COV-2
BUT THEN WITH REMDESIVIR THIS
LIGHTER BROWN IS INHIBITION SO
WE CAN SEE AT PROTEIN LEVEL AS
WELL. WE CAN ALSO DO RNA
SEQUENCING AND LOOK AT A LOT OF
DIFFERENT GENES INHIBITED BY
REMDESIVIR, THAT IS ON THE GREEN
SHOWING THE REMDESIVIR INHIBITED
TYPES OF MESSENGER RNAs. SO
ONE ASPECT OF THIS IS THAT WE
CAN REALLY START TO TAKE THESE
INFECTIONS OUT TO LONGER TIME
POINTS. SO AT EARLY STAGE 3 DAYS
POST INFECTION YOU CAN SEE THERE
IS THE GREEN SARS COV-2 NUCLEIC
ACID SIGNAL, DOESN'T OVERLAP
WITH ANY KIND OF APOPTOTIC
SIGNAL WITH CASPASE 3, JUST RAN
COMELY DISTRIBUTED -- RANDOMLY
DISTRIBUTED TURN OVER WITHIN THE
ORGANOIDS BUT TEN DAYS POST
INFECTION WE CAN SEE SPREAD OF
THE INFECTION AND WITH THE GREEN
NUCLEIC CAPSID SIGNAL BUT NOW
THIS IS OVERLAPPING
SIGNIFICANTLY WITH CLEAVE
CASPASE 3 SO WE ARE STARTING TO
BE ABLE TO TAKE THIS TO AN END
STAGE TYPE OF DISEASE STATE.
THIS IS ANOTHER PICTURE AT TEN
DAYS POST INFECTION. YOU CAN SEE
THE TREMENDOUS SPREAD OF THE
SARS COV-2 NUCLEIC CAPSID SIGNAL
OVERLAPPING SUBSTANTIALLY WITH
THE CLEAVE CASPASE 3. SO AT
THESE LONGER TIME POINTS WE CAN
THEN START TO SEE IMMUNE
CHEMOTACKSIS TO THE INFECTED
AREA, THIS IS JUST THE SAME
IMAGE WITH DIFFERENT STAINS BUT
YOU CAN SEE THAT THE WHITE CD 45
CELLS START TO CHEMO TAX TO AREA
OF ACTIVE INFECTION. AND THEN WE
CAN START TO SEE SOME END STAGE
SEQUELAE. YOU CAN SEE SIN CITY
SHAH FORMATION WHICH IS STRICTLY
OBSERVED IN IF AUTOPSY
SPECIMENS. WITH MULTI-NUCLEATED
CELLS WITH INDICATED BY YELLOW
ARROWS. ALSO WE START TO SEE A
LOT OF INDUCTION OF FIBROTIC
GENES, THIS IS BUNCH OF
DIFFERENT COLLAGEN GENES RNA
SEQUENCING, THIS IS LOG BASE 2
SO ZERO IS ONE FOLD AND SO THERE
IS SUBSTANTIAL INDUCTION THAT
SEEMS TO BE SOMEWHAT HIGHER AT
LATER TIME POINTS. SO
POTENTIALLY VERY INTERESTING FOR
STUDYING PASC TYPE OF SEQUELAE.
FOR SARS COV-2 THEN ON TO T-CELL
AND B CELL RESPONSES ADAPTIVE
RESPONSES WE SHOWED YOU A LOT OF
CYTOKINE INDUCTION MORE INNATE
AND THESE RESPONSES ARE OBSERVED
IN EPITHELIAL ONLY ORGANOIDS BUT
WE WANTED TO TAKE ADVANTAGE OF
HAVING ADAPTIVE IMMUNE
POPULATION T-CELLS AN B CELLS
AND ASSAY THOSE RESPONSES SO
THIS IS A ORGANOID FROM A
IMMUNIZED INDIVIDUAL AND WE DID
INFECTION AND THEN FACT SORTED
CD4 AND CD8 AND JUST SAW IF SARS
COV-2 INDUCE VARIOUS RESPONSES.
THIS IS NORMALIZED SO RATIO MORE
THAN ONE IS INDUCTION AND
THERE'S STRONG INDUCTION OF
INTERLEUKIN 2 AN TNF ALPHA.
THERE'S EVEN INDUCTION OF OTHER
THINGS AS WELL. THAT ARE JUST
OBSCURED BY THE SCALE, LIKE
INTERFERON GAMMA AND THE CD8 WE
CAN SEE CD 68 INDUCTION FOR
INSTANCE, EPITHELIAL CELLS YOU
CAN SEE DEGREE OF SARS GENOME
INCREASE IN EPITHELIUM BUT WE
ALSO SEE INTERFERON IN GCSF FOR
INSTANCE SO INDICATED ADAPTIVE
RESPONSES. WE HAVE ALSO BEEN
ABLE TO STIMULATE THE ORGANOIDS
WITH PEPTIDE MEGA POOLS OR MORE
SIMPLY SARS COV-2 SPIKE PROTEIN,
RECOMBINANT SPIKE PROTEIN. THERE
IS ANTIGEN MACHINERY IN THE
ORGANOIDS I DON'T HAVE TIME TO
TALK ABOUT BUT FEED THE ORGANOID
SPIKE PROTEIN AND OBSERVE VERY,
VERY INTERESTING THINGS LOOKING
AT ORGANOID UNVACCINATED
INDIVIDUALS VERSUS VACCINATED IF
WE FAX PURIFY CD4 AND LOOK AT IL
2 IN THE UNVACCINATED IN THIS
MAYBE GREENISH TYPE OF COLOR,
THERE IS VERY FEW RESPONSES BUT
THEN WITH VACCINATED
PARTICULARLY IF THE PATIENT HAS
BEEN VACCINATED RELATIVELY
RECENTLY, WE SEE HIGHER
INCREASES THAN IF THE PATIENT
WAS VACCINATED MORE IN THE
DISTANT PAST. OF COURSE ALL THIS
PAILS VERSUS THE UNVACCINATED
STATE SO WE ARE LOOKING AT KIND
OF A A RECALL TISSUE RESIDENT
MEMORY IMMUNITY THAT IS
VACCINATION DEPENDENT THAT IS
PRESENT WITHIN THESE LUNG
ORGANOIDS. CD8 SIMILAR THINGS WE
SEE A LOT OF STRONGER RESPONSES
IN VACCINATED VERSUS
UNVACCINATED. AND AGAIN WE CAN
SHOUT THIS OUT BY TIME SINCE
VACCINATION REALLY SUGGESTING
THE TISSUE RESIDENT IMMUNITY
WILL WANE WITH TIME. WE CAN ALSO
SEE SOME DIFFERENCES IN THE --
WITH AGE AS WELL. IF THE
PATIENTS ARE OLDER THAN THEIR
CD4 AND CD8 RESPONSES ARE LESS
VIGOROUS THAN IF THEY ARE
YOUNGER WITH CD4 AND CD8. ON TO
B CELLS, WE SEE THAT SARS COV-2
INDUCES SUBSTANTIAL
IMMUNOGLOBULIN GENE EXPRESSION
THAT IS SPANNING MULTIPLE
ISOTYPES. AGAIN, THIS IS VERY
SUBSTANTIAL INDUCTION OVER THE
BASELINE. AND THEN ALSO IF WE WE
ADD REMDESIVIR, SINGLE RNA
SEQUENCING WITH JUST SARS COV-2
INFECTION WE SEE DOUBLE POSITIVE
CELLS FOR CD 40 AND FOR INSTANCE
IMMUNOGLOBULIN ALPHA IGA GENES
EXPRESSION WHICH IS BEING
INDUCED BUT REMDESIVIR IS REALLY
SHUTTING THIS DOWN. FURTHERMORE
WE CAN LOOK AT SECRETION OF
ANTI-SARS COV-2 SPIKE
IMMUNOGLOBULIN SO WE ARE FEEDING
THE ORGANOIDS THE RECOMBINANT
SPIKE PROTEIN AND ACTIVATING
MEMORY B CELLS AND YOU CAN SEE
THIS IS NOT REALLY -- THIS IS --
THIS IS AN INFECTION SORRY. NOT
FEEDING THE PROTEIN BUT YOU CAN
SEE THERE IS INDUCTION OF
IMMUNOGLOBULIN PRODUCTION
AGAINST SARS COV-2 SPIKE AND WE
CAN SHUT THIS DOWN WITH
REMDESIVIR. SO OF ALL WE HAVE A
HOLISTIC SYSTEM THAT HAS
EPITHELIUM STROMA TISSUE
RESIDENT IMMUNE CELLS, WE CAN
RECAPITULATE SENSITIVITY TO
ANTIVIRAL AGENTS, THE ADAPTIVE
IMMUNE RESPONSES ARE PRIOR SERA
POSITIVITY. WE CAN SHOW THIS
LOCALIZED IMMUNITY WANES OVER
TIME AND LASTLY, THESE ADAPTIVE
RESPONSES, THEY CAN BE INITIATED
AUTONOMOUSLY IN LUNG BECAUSE
LUNG ORGANOIDS DO NOT HAVE
PERIPHERAL LYMPHOID COMPONENT SO
WE CAN SHOW WE HAVE SUFFICIENCY
NECESSARY WITHIN THE LUNG FOR
ANTIGEN, INFECTION ANTIGEN
PRESENTATION, AND ADAPTIVE
MEMORY RESPONSES. SO I WILL
STOP THERE AND THANK YOU VERY
MUCH.
>> FANTASTIC, DR. KUO.
APPRECIATE THAT TOUR DEFORCE
PRESENTATION. IF I CAN ASK ALL
SIX PANELISTS TO JOIN US ON
SCREEN, I THINK IT WOULD BE
GREAT TO KICK OFF THE Q&A
SESSION. THANKS FOR JOINING ON
VIDEO KEEPING US COMPANY. I
GUESS MAYBE I WILL GO TO -- A
QUICK REMINDER, PLEASE ENTER ANY
QUESTIONS YOU MIGHT HAVE FOR ANY
OF OUR SPEAKERS IN THIS SESSION
INTO THE Q&A BOX AND WE WILL GET
TO THEM. PLEASE NOTE THAT OUR
SPEAKERS WERE VERY GENEROUS AND
EFFICIENT IN ANSWERING SOME OF
THE QUESTIONS THAT WERE POSTED
WILL IN WRITING SO BE SURE TO
FLIP TO ANSWERED TAB TO LOOK AT
THE RESPONSES IF YOU HAVE ANY
FOLLOW-UP OR OTHER QUESTIONS
RAISED PLEASE FEEL FREE TO TYPE
IN A NEW QUESTION AND WE WILL
CIRCLE BACK TO THAT. JUST WANT
TO HIGHLIGHT THE DIFFERENT TABS
IF YOU WILL FROM THAT Q&A BOX.
TO KICK OFF I WILL LOOK THROUGH
WHAT IS IN THAT BOX FOR OPEN
QUESTIONS. ONE THIS IS FOR DR.
INGBAR, THANK YOU FOR THE TALK,
IT SEEMS THAT USING CHIPS FOR
EFFICACY AND REPURPOSING WILL --
WHERE THERE ARE EXISTING SAFETY
DATA MAYBE LOW HANGING FRUIT.
MORE LIKELY TO BE ACCEPTED FOR
REGULATORY SUBMISSIONS. CAN YOU
SEE CHIP DATA BEING USED IN LIEU
OF ANIMAL DATA FOR TESTING
TOXICITY OR DO WE NEED TO
RETHINK WHAT SAFETY QUESTIONS WE
ARE ASKING AND WHAT ANSWERS WE
NEED.
>> YEAH. WE HAVE A PAPER IN
REVIEW, IN PRE-PRINT WITH
EMULATE COMPANY THAT MAKES THESE
CHIPS, WHERE THEY DID 800
DIFFERENT HUMAN LIVER CHIPS THAT
ARE ALIGNED BY PRIMARY HUMAN
HEPATOSITES PRIMARY LIVER
ENDOTHELIUM PRIMARY HUMAN AND
THEY TESTED 27 DRUGS. FOR DRUG
INDUCED LIVER INJURY. DRUGS
LISTED BY THE IQ CONSORTIUM
WHICH IS CONSORTIUM OF PHARMA
AND BIOTECH COMPANIES THEY
IDENTIFY THEY KNOW THE RESULTS
IN ANIMALS AND MOW THE RESULTS
IN HUMANS IN ABOUT 18 OF THOSE
ANIMALS WERE SAFE BUT THERE WAS
TOXICITY IN HUMANS AND THE NET
SUM WAS 780 CHIPS BUT NOW 800
WITH TWO DONOR, A THIRD DONOR.
80% SENSITIVITY AND 100%
SPECIFICITY. THAT IS 8 TIMES
BETTER THAN ANIMAL MODEL. SO WE
ARE HOPING THAT PEOPLE WILL
START USING THINGS LIKE THAT FOR
TOXICITY AS ONE EXAMPLE. THERE
IS ANOTHER COMPANY DOING KIDNEY
TOXICITY. BLANKING THE NAME OF
THE COMPANY BUT FOUND IT IS ALSO
MUCH BETTER THAN ANIMAL MODELS.
SO WE ARE APPROACHING A TIPPING
POINT BUT WE ARE NOT THERE YET.
>> GREAT. THANK YOU. SO NEXT
QUESTION FROM THE BOX. AGAIN FOR
DR. INK BEAR. HAVE YOU DEVELOPED
LUNGS ON CHIPS FOR CYSTIC
FIBROSIS PATIENTS? AND HAVE YOU
TRIED TO EPIIF HE CAN THOSE WITH
BACTERIAL PATHOGENS?
>> WE PUBLISHED LAST YEAR WORK
FUNDED BY CYSTIC FIBROSIS
FOUNDATION, WE GOT AIRWAY CELLS
FROM PATIENTS AND THE CHIPS
TOTALLY WE MAKE THE PHENOTYPE
INCREASE CILIA, THICKENED
MUCOUS, ALTERED EATING
FREQUENCY, INCREASE BASELINE
INFLAMMATORY STATE INCREASE
RECRUITMENT OF IMMUNE CELLS AND
WE GREW SINEMONAS. JOE THAT IS
PUBLISHED.
>> THANK YOU. FOLLOW-UP, CURIOUS
WHETHER YOU -- I THINK YOU HAD
SHOWN IN YOUR TALK YOU LOOKED AT
THAT TIME COPD SAMPLES IN
COMPARE TO HEALTHY. WONDERED IF
YOU LOOK IN TERMS OF VIRAL
INFECTION OR PATHOGENESIS AND
WONDERING IF YOU LOOKED AT THAT
SMOKING IN PARALLEL WITH HEALTHY
TO SEE IF THERE'S ANY IMPACT ON
VIRAL INFECTIVITY OR
PATHOGENESIS.
>> IT IS A MUCH MORE COMPLEX
MODEL SYSTEM TO GET WORKING AND
WE DID THAT A COUPLE OF YEARS
AGO SO WE HAVE BE INTERESTING TO
SEE BUT WE HAVEN'T DONE THAT
ONE.
>> THANKS. NEXT I SEE HERE IS
FOR DR. KUO. THE QUESTION IS CAN
THESE MORE COMPLEX LUNG
ORGANOIDS BE EXPANDED AND
CRYOPRESERVED LIKE THE MORE
CONVENTION PAL EPITHELIAL ONLY
ORGANOORGANOIDS?
>> I THINK SO. WE HAVEN'T TRIED
THIS FOR THE MORE COMPLEX LUNG
ORGANOIDS. FOR ANALOGOUS TUMOR
ORGANOIDS IT IS POSSIBLE. SO WE
ARE STARTING TO DO THIS WHAT WE
SUGGEST MARK AND WE DON'T HAVE A
LOT OF DATA YET BUT THINK IT IS
POSSIBLE.
>> NEXT FOR DR. KUO. POLEDOR
WOULD YOU ELABORATE HOW YOU
SOURCE YOUR IMMUNE SPHEREROIDS
IN CULTURE THEM IN THE 3D ALI
ENVIRONMENT PRIOR TO
CHARACTERIZATION AND
PERTURBATION FROM THE MODEL? ARE
THEY REMOVED FROM PATIENT
BIOPSIES, HOW LONG TO MAINTAIN
INFECTION?
>> EVERYTHING IS FROM PATIENT
BIOPSIES BECAUSE THESE ARE
THEMSELVES CELL DIRECT
ORGANOIDS.
SO TYPICALLY SURGICAL RESECTIONS
FROM NORMAL MARGIN OF CANCER
PATIENTS OR FROM SOMETIMES
CHILDREN THERE IS VARIOUS TYPES
OF BIRTH DEFECTS FOR WHICH TH TE
IS A LUNG RESECTION THEN A
NORMAL REGION ASIDE FROM THE
EFFECT IS. AND RIGHT. WE CULTURE
ESTABLISH ORGANOIDS FIRST TO
MAKE SURE THEY ARE GROWING THEN
WE WILL FLIP THEM IN CULTURE TO
GET FOR INSTANCE THE ACE 2 TO
THE OUTSIDE THEN WE INFECT.
>> GREAT. THANK YOU. SO THE NEXT
QUESTION IS FOR ANYONE, PLEASE
FEEL FREE TO SHOUT OUT IF YOU
HAVE A THOUGHT OR IDEA. SO TO
ALL THE SPEAKERS ARE YOU LOOKING
AT POPULATION VARIABILITY AND
SUSCEPTIBILITY TO ENVIRONMENTAL
CHEMICALS AND HOW THAT MIGHT
INFLUENCE THE VIRAL INFECTIONS
OR VICE VERSA? ANY THOUGHTS ON
THAT FOR THE SPEAKERS?
>> WE ARE DEFINITELY LOOKING AT
POPULATION VARIABILITY NOT IN
THE CONTEXT OF CHEMICAL EXPOSURE
AT THIS POINT IN TIME BUT LOOK
AT DIFFERENT DONORS WHETHER FROM
CHILDREN OR ADULTS DEPENDING
CONTEXT OF WHAT YOU ARE
STUDYING. FOR EXAMPLE IN OUR
CASE OF NOROVIRUS INFECTION WE
LOOK AT SECRETERS NON-SECRETERS,
SO DEFINITELY CONSIDERING
POPULATION HETEROGENEITY BUT NOT
IN THE CONTEXT OF CATEGORY
(INAUDIBLE).
>> NO, THANK YOU FOR THAT. I DID
REMEMBER IN YOUR TALK REMEMBER
CROSSED MY MIND, THERE MUST BE
-- IT MUST BE CHALLENGING TO GET
THAT BALANCE BETWEEN WANTING TO
SAMPLE THE BROAD POPULATION BUT
AT THE SAME TIME GENERATE THAT
SORT OF REPRODUCIBILITY FOR
SCREENING AND SO THOUGHT THAT
WAS INTERESTING POINT YOU RAISED
IN THAT REGARD.
>> I THINK IT IS POINT IT IS
LARGELY A CONVENIENCE, WE ARE
GETTING WHAT WE ARE GETTING OR
AT LEAST WHERE WE STARTED, MUCH
MORE MEDIAN SAMPLE, IT IS ONE OF
THE LINES WE HAVE GOT
CHARACTERIZED THEM AND DIVIDED
THEM INTO THE POPULATIONS WE
KNOW ARE RELEVANT FOR WHAT WE
ARE STUDYING. ASKING MORE
FORWARD WE ARE NOW GOING INTO
MORE SPECIFIC DISEASE. WE WANT
TO STUDY THIS AND THEREFORE
WHICH GO TO WHATEVER ORGANOIDS
THAT ARE NECESSARY FOR AN AGE
GROUP OR PARTICULAR GENETIC
SUSCEPTIBILITY FACTOR. WE TAKE
THAT INTO CONSIDERATION.
>> ONE THING THAT IS ALWAYS
BROUGHT UP WITH ORGANOIDS AND
ORGANS ON CHIPS IS VARIABILITY
ISSUES BECAUSE YOU DON'T HAVE
ESTABLISHED CELL LINE OR
ESTABLISHED ANIMAL MODEL BUT TO
ME THAT IS THE POWER, THAT IS
WHAT REAL CLINICAL TRIALS IS
ABOUT IS PATIENT TO PATIENT
VARIABILITY AND SO FORTH. SO WE
FOUND THAT WE USUALLY DO
MULTIPLE DONORS LIKE YOU HEARD.
THAT IS ONE AREA. BUT WE HAVEN'T
DONE IT FOR VIRAL INFECTION BUT
WE HAVE OTHER GRANTS WE ARE
WORKING ON FOR EXAMPLE VAGINA
AND CERVIX MODELS OF THE
MICROBIOME AND WE MADE CAUCASIAN
AFRICAN AMERICAN AND HISPANIC
AND THEY ARE DIFFERENT WHICH IS
INTERESTING. WE HAVE A GRANT ON
RADIATION COUNTER MEASURES FROM
FDA DOING MALE VERSUS FEMALE
BECAUSE IN CLINICAL TRIALS, NOT
COVERED EQUALLY IN A DIFFERENT
-- SO THAT IS A POWER OF THESE
BUT IT MAKE MORE DIFFICULT
BECAUSE YOU HAVE TO DO MORE
VARIABLES.
>> I WANT TO ASK A QUESTION TO
FOLLOW-UP ON THAT. THERE WAS A
RESPONSE FOLLOW-UP TO QUESTION
DR. INGBAR RESPONDED TO. WHICH
IS IF YOU ARE WORKING WITH DRUG
INDUCED DISEASE AND YOU SAY YOU
HAVE THREE DONORS YOU INVESTED
GIVEN YOUR WORK WOULD SAY FDA --
IS THERE A CERTAIN EXPECTATION
THAT YOU SHOULD HAVE SIX DONORS
OR TEN DONORS, WHEN ARE WE
GETTING TO THAT NUMBER AS A
FIELD IN GENERAL?
>> MY FIRST INTERACTION WITH THE
FDA PRETTY CONSISTENTLY EARLY
ON, FROM THOSE WHO ARE OPEN TO
THIS STUFF IS AS LONG AS YOU ARE
AS GOOD AS AN ANIMAL MODEL, OR
BETTER, WEIAL STATISTICAL
ROBUSTNESS AND IT IS HUMAN, THEY
REVIEW THAT AS A POSITIVE.
BECAUSE REALLY YOU ARE DEALING
WITH COMPETING WITH ANIMAL
MODELS WHERE THEY TOTALLY INBRED
YOU GET DIFFERENT RESULTS WITH
DIFFERENT BREEDS SO BUT THERE IS
NO DOUBT ONCE YOU START USING
THESE MODELS THE BIGGER
QUESTIONS COME UP. NOW -- CAN
YOU ONLY USE IT FOR MEN, OR CAN
YOU ONLY USE IT FOR CERTAIN AGE
GROUPS? SO THE FDA IS STILL NOT
CLEAR HOW TO HANDLE ALL THESE,
THEY ONLY DEAL WITH SAFETY. SO
IT IS DIFFERENT THAN THE
PHARMACEUTICAL BIOTECH INDUSTRY.
FROM THE CLINICAL SIDE THEY ARE
DIFFERENT CHALLENGES, YOU MIGHT
WANT PERSONALIZED MEDICINE SO IT
IS A LITTLE BIT UP IN THE AIR
BUT I DO THINK FOR SAFETY YOU
ARE COMPETING WITH ANIMAL MODELS
AND AS LONG AS IT IS HUMAN AND
IT IS -- YOU CAN REPLICATE
CLINICAL MIMICRY WITH THE MODEL,
THAT IS WHAT WE USE, DO WE SEE
THIS IN VIVO IN HUMANS CELL
POPULATIONS TOO. TO WHAT THAT
VALIDATION IS. HOW ARE YOU
VALIDATING THE MODEL SO IT IS
PROBABLY RELATIVE MAYBE A LITTLE
BIT NARROWER AREA LIKE YOU ARE
SAYING.
>> SO ANOTHER QUESTION ON THE
BOX FOR SARA. TERRIFIC
PRESENTATION SARA WITH REGARD TO
THE NASAL VERSUS AIRWAY
PLATFORMS DO YOU SEE A
SYSTEMATIC DIFFERENCE IN VIRAL
REPLICATION RATES SUGGESTING
MORE RAPID REPLICATION IN LUNG
TISSUE VERSUS THE OTHER?
>> WE SEE SARS COV-2 REPLY CASE
ROBUSTLY IN BOTH MODELS THOUGH
WHEN COMPARING ACROSS THE
VARIANTS THERE ARE SOME
DIFFERENCES IN PATHOGENESIS IN
THE DIFFERENT CULTURE SYSTEMS
WHERE THERE IS MORE TOXICITY FOR
VARIANTS IN ONE MODEL OR THE
OTHER. THEN IN TERMS OF THE DRUG
TREATMENT THERE IS CERTAINLY
DIFFERENCES IN EFFICACY BETWEEN
DIFFERENT CLASSES OF SMALL
MOLECULES, AND WHAT THAT MEANS
IS LESS CLEAR TO US BUT WE ARE
DOING EXPERIMENTS IN ANIMAL
MODELS COMPARING EFFICACY IN
NASAL EPITHELIUM VERSUS THE
LOWER RESPIRATORY TRACT SO WE
DON'T KNOW IF WE ARE MODELING
WELL OR NOT BUT THERE ARE
CERTAINLY DIFFERENCES BETWEEN
DIFFERENT TISSUES THAT MAY
REFLECT REALITY, JUST NOT SURE
YET BECAUSE WE HAVEN'T BEEN AS
SYSTEMATIC AS WE NEED TO BE.
>> THANK YOU. THERE IS A TOPIC
IN THE ANSWER COLUMN NOW THAT
WENT ON IN THE CHAT THAT I
THOUGHT WAS REALLY WORTH
BRINGING UP UP. I THINK SASHA YU
RAISED IT BUT IT RAISES I GUESS
A QUESTION, I WILL READ YOUR
QUESTION SO YOU WORD IT BETTER
THAN I WAS TRYING TO. AND THAT
WAS TO DR. LEE TO KICK OFF THE
DISCUSSION BUT I THINK I WOULD
LIKE TO HEAR OTHERS CHIME IN AS
WELL. THE QUESTION OR COMMENT
WAS CURIOUS TO LEARN MORE ABOUT
YOUR THOUGHTS ON CHARACTERIZING
THE MODELS EARLY AND WHAT KIND
OF CHARACTERIZATION DO YOU THINK
IS IMPORTANT TO BE THOROUGH
ENOUGH. AGREE WOULD BE GREAT TO
HAVE WELL CHARACTERIZED MODELS
READY TO BE USED FOR DIFFERENT
INFECTIONS AND DISEASES, THIS
DEFINITE CAME UP IN MY THINKING
AS WELL AROUND THE SORT OF
PANDEMIC PREPAREDNESS MORE
BROADLY AND THINKING ABOUT HOW
DO WE PREPARE FOR THAT PATHOGEN
X REFLECTING BACK TO WHERE WE
WERE WITH SARS COV-2 WE ARE
FORTUNATE ACTUALLY THE
FOUNDATIONAL STUDIES WITH SARS
AND OTHER COMA VIRUSES THAT GAVE
CLUES WHETHER CELLS LINES TO USE
AND WE BUMPED ALONG SETTING UP
ASSAYS AND MODELS AND UNDERSTAND
CELL LINES BUT LOVE TO BE ABLE
TO DO IS INVEST MORE EFFORTS
DURING PEACETIME IF YOU WILL
ACROSS THE VIRAL FAMILIES THAT
MAY HAVE RELATED VIROS BE FUTURE
PROBLEM AND UNDERSTAND BASIC
FOUNDATIONAL WORK IN THAT SPACE
SO THAT WE A WE ARE BETTER TO
ACCELERATE RAPIDLY WHEN WE SEE A
PATHOGEN HIT OR RADAR. SO DR.
LEE IF YOU COULD KICK OFF EMILY
AND SHARE YOUR THOUGHTS HOW YOU
ARE SETTING UP TO TO PREPARE
FROM THAT AND LOVE TO HEAR
OTHERS HOW WE CAN DO THIS. FOR
THIS EXACT PURPOSE SO WE ARE
WORKING THROUGH DIFFERENT
MODELS, DIFFERENT VIRAL FAMILY,
DIFFERENT REPRESENTATIVE VIRUSES
IN THOSE FAMILIES AND STARTING
FROM THE VERY BASICS, STARTING
WITH WHAT ARE THE VIRAL KINETICS
IN THAT TISSUE WHAT IS CELLULAR
TROPISM IN THAT TISSUE, ARE WE
OBSERVING VIRAL PERSISTENCE,
LOOKING IN PARTICULAR IN THE GUT
FOR SARS COV-2, CAN WE ESTABLISH
PARAMETERS WHICH IS NOT EASY
CONTROL ANTIVIRAL HERPETIC FOR
BENCHMARKING FOR EVALUATING NEW
THERAPEUTICS IF POSSIBLE. THEN
IN TERMS OF VIRAL DISEASE WHICH
IS ALSO IMPORTANT TO LOOK AT WE
REALLY WANT TO BE ABLE TO
CHARACTERIZE TISSUES IN TERMS OF
HEALTHY UNINFECTED TISSUE VERSUS
VIRAL INFECTED TISSUE TO GET
IDEA WHAT READ OUTS WE SHOULD BE
USING TO EVALUATE THERAPEUTIC
EFFICACY BOTH DIRECTLY LOOKING
AT VIRAL INFECTION BUT THEN ALSO
AFFECTING THE TISSUE. SO WE ARE
TRYING TO LOOK AT MORPHOLOGICAL
READ OUTS, WE HAVE RECENTLY
STARTED USING SINGLE CELL RNA
SEQ SO WE HOPE TO MAKE THAT DATA
PUBLIC, AS WE GENERATE AND ALSO
HAVING FUNCTIONAL ASSAY AS WELL.
SO THIS IS A HUGE PROJECT THERE
IS A LOT OF PEOPLE INVOLVED AND
ALWAYS WELCOME COLLABORATORS TO
WORK WITH AS WELL TO TRY TO GET
THIS INFORMATION OUT FOR GENERAL
USE.
>> I WAS GOING TO JUMP IN, THIS
IS A REALLY IMPORTANT QUESTION
HOW WE PREPARE FOR THE NEXT
PANDEMIC WITH THESE MODELS. SO
IN OUR LABORATORY WITH WE ARE
INTERESTED IN THE QUESTION THAT
WAS ASKED TO ME BY MARK EARLIER,
WE WANT TO ESTABLISH BIOBANKS IN
THE ORGANOIDS THAT ARE
REPRESENTING DIFFERENT AGES
ETHNICITIES AND IN THE CASE OF
SARS VACCINATION STATUS, BECAUSE
WHEN WE SURVEY DIFFERENT
ORGANOIDS FOR ADAPTIVE IMMUNE
RESPONSE WE SAW VERY DIFFERENT
RESPONSES WHETHER PATIENT DONOR
HAS BEEN VACCINATED PREVIOUSLY
OR NOT. SO THAT AUTOMATICALLY
SHOWS TREMENDOUS INTERPATIENT
VARIABILITY DEPENDING CERTAIN
CONCEPTS SO HAVING THESE
BIOBANKS AVAILABLE THAT SPAN
AGES ETHNICITY, VACCINATION
STATUS, ALONG WITH BLOOD BANKED
IN PARALLEL IS GOING TO BE VERY
IMPORTANT. SO THAT IS DEFINITELY
SOMETHING WE ARE INTERESTED IN
TRYING TO PURSUE WORKING WITH
Y'ALL.
>> SO WHAT ONE THING I WOULD
ALSO MENTION IS THAT FOR MANY
EMERGING VIRUSES, WE DON'T KNOW
WHAT TROPISMS ARE THE VIRUSES
WHAT CELLS GET INFECTED. AN
EXAMPLE IS TICK BORNE VIRUSES
BECOMING INCREASINGLY WORRISOME
BUT WE REALLY DON'T KNOW
ANYTHING ABOUT WHAT CELLS GET
INFECTED. THAT IS A HUGE
OPPORTUNITY TO USE STEM CELL
DERIVED ORGANOIDS TO MAKE
VARIOUS ORGANOIDS AND TEST THEM.
FOR MANY ARTHEROPOD BORNE
VIRUSES DENDRITEICS DERMAL
DENDRITIC CELLS ARE MOST
IMPORTANT BUT THESE VIRUSES PAST
THROUGH OTHER TISSUES ON THE WAY
TO THE BRAIN. THERE IS FROM
TREMENDOUS OPPORTUNITY TO USE
STEM CELL DERIVED CELLS TO LOOK
AT EACH OF THESE DIFFERENT CELL
TYPES AND SEE OTHER DIFFERENCES
IN REPLICATION, DO VIRUSES
CHANGE, THIS KIND OF THING. SO A
LOT OF GOOD THINGS COULD BE
DONE.
>> GREAT. ONE THOUGHT MAYBE YOUR
EXPERIENCE AND LOOKING ACROSS
THE BOARD AT DIFFERENT CELLS
DIFFERENT COMPLEXITY OF MODEL
ASSAYS, ONE OF THE THOUGHTS I
WAS HAVING IS FOR THAT UNKNOWN
PATHOGEN OR SURPRISE PATHOGEN
THAT COMES IN STARTING WITH MORE
COMPLEX MODEL ASSAYS BUT IT
MAKES SENSE AS YOU WERE SAYING
DR. GEHRKE BUT WONDERING IF
UNDERSTANDING THOSE SYSTEMS
ALLOW US TO THEN RETROSPECTIVE
BACK UP TO MORE SIMPLE CELL LINE
AND UNDERSTAND WHICH CELL LINES
WOULD BE BEST FOR WHAT TYPE OF
VIRAL FAMILIES AND THAT KIND OF
THING. AND WONDERING IF THERE IS
FOUNDATIONAL WORK TO BUILD THAT
TRANSLATION BETWEEN COMPLEX
SYSTEMS LEADING BACK TO VARIETY
OF MORE SIMPLE CELL ASSAY AND IF
THERE IS VALUE IN THAT OR IS
THAT DEFEATING THE PURPOSE OF
WHAT WAS GOING THROUGH MY MIND?
>> WE WORK WITH A LOT OF
DIFFERENT RNA VIRUSES IN MY LAB
AND WHAT WE HAVE BEEN TRYING TO
DO NOW IS DEVELOP CELL LINE
BASED MODEL CLOSE AS POSSIBLE TO
RELEVANT CELL TYPES IN VIVO
ASSAYS FOR THEM. SO WE HAVE
ASSAYS FOR HALF A DOZEN
FLAVIVIRUSES AND CORONA VIRUSES
AND MIX OF VIRUSES, IN A NUMBER
OF DIFFERENT CELL TYPES THAN WE
THINK ARE NOT VIRILE CELLS AND
NOT 3D MODELS BUT SOMETHING IN
BETWEEN TO REFLECT TISSUES THAT
ARE INFECTEDED. SO WE HAVE BLOOD
BRAIN BARRIER ENDOTHELIAL CELLS
VERSUS OTHER ENDOTHELIAL CELLS
AND THESE 3s ARE A GOOD MODEL
FOR THE RESPIRATORY EPITHELIUM
SO WE ARE TRYING TO COME UP WITH
A TOOLBOX DIFFERENT CELL TYPES
WE HAVE HIGH THROUGH PUT ASSAYS
SETS UP FOR DIFFERENT VIRUSES WE
HAVE HIGH THROUGH PUT ASSAYS SET
UP TO MIX AND MATCH RAPIDLY
BECAUSE WHEN SARS CAME OUT WE
TESTED EVERY CELL LINE WE CAN
BUY FROM ATCC AND THEN ENDED UP
ON CALU SO HOPEFULLY WE CAN HAVE
COUPLE OF HUNDRED CELL LINES IN
THE LAB AND FIGURE OUT WHAT IS
THE BEST MODEL FOR CELL TYPE IS
IMPORTANT. NEVER GOING TO BE THE
SAME AS 3D MODELS BUT AS SOMEONE
WHO IS A USER NOT A DEVELOPER,
IT IS HARD TO GET ACCESS TO
THESE MODELS, DIFFERENT LABS
DIFFERENT MODEL AND I'M NOT SURE
WHAT IS BETTER OR WORSE BETWEEN
THEM ACTUALLY WHICH IS PARTLY
WHY WE USE DIFFERENT MODELS FROM
DIFFERENT SOURCES, BUT I THINK
WE HAVE A LOT TO LEARN OR IF ALL
WE CAN DO IS START WITH 3D
MODELS IT IS HARD TO DO HIGH
THROUGH PUT SCREENING SO WE NEED
SOMETHING IN BETWEEN (INAUDIBLE)
>> GREAT. THANK YOU. ANY OTHER
THOUGHTS ON THAT? BEFORE SHIFT
GEARS A LITTLE BIT?
>> MAY I ASK A FOLLOW-UP
QUESTION?
>> PLEASE.
>> I WAS WONDERING, IF SARS
COV-2 IS A PRIME EXAMPLE FOR THE
MISSION WHERE YOU HAVE YOUR YOU
HAVE THE DIFFERENT CELL LINES,
YOU HAVE THE CELLS YOU HAVE 3D
MODELS AND CLINICAL EFFICACY
DATA SO ER TROUGH SPECKTIVELY
THINK HERE -- RETROSPECTIVELY
THINK HERE IS WHERE WE GET THE
CORRELATION AND USE THE
KNOWLEDGE TO INFORM US FOR THE
FUTURE. DO YOU ENVISION
SOMETHING LIKE THAT BASED ON
YOUR EXPERIENCE WITH SARS COV-2?
>> I PERSONALLY THINK IN TERMS
OF THE CELL LINES THE CALU 3
CELLS ARE CLOSEST TO A LUNG BUT
THEY ARE NOT A LUNG SO BACKWARDS
I SHOWED YOU DATA MAYBE NOT VERY
CLEAR, THIS WAS ONLY 20 MINUTES
BUT WE HAVE DATA THREE APPROVED
DRUGS REMDESIVIR AND PAXLOVID
HAS ACTIVITY IN ALL THE MODELS
WE TESTED. NOT ALL CELL LINES
BUT IN VIVO AND IN CALU SO I
THINK FOR THAT CLASS OF VIRUSES
I THINK A GOOD IN BETWEEN MODEL.
BUT FOR TRYING TO DEVELOP THESE
FOR OTHER VIRUSES, SO I THINK
FOR OTHER RESPIRATORY VIRUS VIRS
CALU IS GOOD BUT WE ARE
DEVELOPING THEM WHAT WE THINK
ARE AS GOOD AS THAT FOR OTHER
TISSUE TYPES AND I DON'T KNOW
THERE WILL BE ONE FOR EACH
TISSUE TYPE EVEN
>> I GUESS ANOTHER SORT OF AREA
TOPIC --
>> CAN I --
>> PLEASE DO.
>> I THINK FOR RAPID SCREENING
IT MAKES SENSE CALU 3 OR PEOPLE
NOW WORKING ON AUTOMATING
ORGANOIDS WHICH MAYBE BETTER.
SO COULD BE ANOTHER WAY THE GET
TO NEXT STAGE. FROM OUR WORK WE
WERE LOOKING MORE HOST RESPONSE
RATHER THAN JUST GETTING VIRUS
TO GROW AND VIRUS INFECTION
MECHANISMS AND SO FORTH. JUST
BECAUSE WHEN YOU THINK ABOUT THE
EARLY DAYS OF COVID-19, IT WAS
REALLY THE CYTOKINE STORM IN THE
INFLAMMATORY RESPONSE THAT IS
KILLING PEOPLE. NOT THE INITIAL
INFECTIONS. SO YOU HAVE TO KEEP
THAT IN MIND TOO. WE NEED BOTH
ABSOLUTELY NEED BOTH. THAT IS
WHERE MORE COMPLEX MODELS COME
IN TRYING TO FIND HOST --
ANALYZE HOST RESPONSE AND FIND
MODULATORS.
>> I AID GROW AND I REALLY
PERKED UP IN YOUR PRESENTATION
WHEN YOU WERE TALKING ABOUT I
THINK IT WAS THE RAGE INHIBITOR
THAT YOU PROFILED BECAUSE I
THINK TO THIS DAY WE HAVE A
COMPLETE GAP IN ABILITY TO
PRE-CLINICALLY ASSESS CANDIDATE
DRUGS FOR THAT SORT OF PHASE OF
DISEASE. AND THERE ARE A FEW --
WE WERE BASICALLY DOING OUR
ANIMAL MODELS WHO ARE IN
CLINICAL TRIALS FOR THOSE DURING
THE SARS COV-2 PANDEMIC FOR GOOD
REASON, THAT'S WHERE PEOPLE WERE
REALLY SUFFERING AND WE NEEDED
INTERVENTIONS. I THINK THAT WILL
BE NOT AN UNCOMMON SITUATION TO
BE IN FOR RESPIRATORY VIRAL
INFECTION. SO JUST LOVE TO HEAR
MORE ABOUT HOW MUCH WORK YOU
HAVE DONE IN THAT SPACE AND
PROFILING SOME OF THE KNOWN
CLINICAL CANDIDATES OR CLINICAL
DRUGS HITTING THAT MORE ARDS
TYPE PHASE OF INFECTION, HAVE
YOU LOOKED AT THOSE AND IS YOUR
SYSTEM RECAPITULATED IN CLINICAL
DATA THERE?
>> OUR FUNDING FROM DARPA WAS
ONE YEAR AND ENDED A YEAR AGO.
COMING TO AN END BUT THAT WAS ON
INFLUENZA FOR THE MOST PART. SO
WE HAVEN'T BUT YOU ARE
ABSOLUTELY RIGH RIGHT. WE HAVE
UNCOVERED A LOT OF INTERESTING
INSIGHTS INTO THE MCNISTIC
UNDERPINNINGS OF THAT --
MECHANISTIC UNDERPINNINGS OF
THAT BUT ONCE WE PICK THAT UP
ABOUT THE RAGE RECEPTOR YOU
START LOOKING AT THE LITERATURE
OUT THERE IT IS OVERWHELMINGLY
INDICATIVE IT PLAYS A CENTRAL
ROLE IN THIS PROCESS. I THINK
THIS IS COUNTERPOINTING OF RAPID
SCREENING, FOR JUST GOT TO
UNDERSTAND SOMETHING ABOUT THE
VERDICT YOU HAVE TO PROPAGATE
THE VIRUS TO STUDY THE VIRUS.
ABSOLUTELY NEED TO FIND WHAT
CELL LINES ARE RIGHT FOR THAT
AND THEN THE INITIAL ENTRY AND
DYNAMICS AND HOST RESPONSE WE
NEED ALL THOSE BUT CLOSER WE GET
TO HUMAN THE BETTER BECAUSE I
NOCIMON WILL PROBABLY TALK AT
THE END BUT JUST ANIMAL MODELS
WERE NOT OPTIMAL IN THIS ONE.
>> AGREE. WE ARE GETTING CLOSE
TO THE END BUT I WANTED TO PULL
-- ASK A FOLLOW-UP A LITTLE BIT
MORE ON I THINK SASHI YOU
MENTIONED IN YOUR TALK HAS A
NICE STORY HOW IT WAS IMPORTANT
TO IDENTIFY WHERE YOU WERE IN
THE END POINT ASSESSMENT AND HOW
SENSITIVE YOU WOULD BE TOWARD
TOXICITY OR CYTOTOXICITY TO GET
YOUR AND SENSITIVITY OF ASSAY TO
GET RELIABLE RESULTS. NICE
STORY, I WONDER FOR ALL THESE
SYSTEMS HOW BIG ISSUE ASSESSMENT
OF CYTOTOXICITY IS BECAUSE THAT
CERTAINLY IS AN ISSUE DOING
SIMPLE CELL SCREENING AND NOT
ONLY SURE YOU CAN DO A CYTOSOCK
AT THISTY SCREEN FOR THAT OVERT
CYTOTOX BUT MORE SUBTLE
INTERFERENCE OF CELLULAR
METABOLISM, THEREFORE GIVING YOU
A READ THAT LOOKS LIKE REDUCING
VIRAL LOAD I THINK IS AN
ARTIFACT THAT COMES UP ALL THE
TIME AND CELLS SCREENING AND I
WOULD IMAGINE THAT COULD BE ALSO
MAYBE EVEN MORE PROBLEMATIC IN
THESE MORE COMPLEX SYSTEMS,
WONDER WHAT FOLKS THINK OF THAT
STARTING WITH YOU TO ADDRESS
YOUR EXPERIENCE IN THAT SPACE.
>> IT WAS A SURPRISE TO US TO
SEE HOW MUCH VARIABILITY WAS
THERE AND HOW STANDARD PROTOCOL,
ANTIBODY USE OF PROTOCOL REALLY
DIDN'T WORK AS WELL IN OUR
HANDS. SO I DON'T THINK IT IS
REALLY WE ARE AT THE BEGINNING
OF THE LEARNING CURVE FOR THESE
THINGS AND CERTAINLY FOR US WE
APPRECIATE EACH OF OUR LINES ARE
QUITE DIFFERENT, THERE'S SOME
LINES WHICH ARE VERY DIFFERENT
FROM THE OTHERS, FOR EXAMPLE. SO
DOING TRANSCRIPTIONAL ANALYSIS
FOR EXAMPLE TRYING TO FIGURE OUT
WHAT IS DIFFERENT ABOUT THESE
LINES, IN SOME CASES WE HAVE
BUNCH OF SEQUENCE THE GENOME SO
THERE ARE DIFFERENCES IN DRUG
METABOLIZING ENZYMES IN GENERAL,
THOSE ARE ALL THE PLACES WHERE
WE ARE GOING TO BEGIN TO GET
ANSWERS WE DON'T HAVE PERMISSION
TO SEQUENCE GENOME FOR ALL OF
OUR CULTURES BUT I THINK THAT IS
WHERE WE ARE IN TERMS OF TRYING
TO UNDERSTAND WHY THERE ARE
DIFFERENCES. ANY OTHER
PANELISTS HAVE EXPERIENCE IN
TRYING TO SORT THROUGH CYTOTOX
VERSUS ANTIVIRAL EFFECTS IN
THEIR SCREENING EFFORTS?
>> ONE NICE THING ABOUT ORGAN
CHIPS IS WE CAN MEASURE
PHYSIOLOGICAL TOXICITIES LIKE
TISSUE BARRIER INTEGRITY,
CILIARE MOTION P PRODUCTION,
CYTOKINE PRODUCTION SO FORTH. SO
WE GET A LOT OF INFORMATION ON
THE SYSTEM MORE THAN JUST CELL
INJURY THE CYTOTOXICITIES ARE
RELATED TO PHARMACOKINETICS MOST
STATIC SYSTEMS IS BATHED IN
DRUGS FOR EXTENDED TIMES SO AS I
MENTION WE TRY TO USE WE USE C
MAX IN THESE STUDIES AT STEADY
RATE BUT WE MIMIC KNOWN PK
PROFILES IN OTHER PAPERS AND
THAT I THINK IS THE FUTURE.
>> IT IS REALLY IMPORTANT TO
COME UP WITH GUIDELINES FOR
PEOPLE IN TERMS OF REPORTING AND
IN TERMS OF WHAT WE EXPECT. WE
HAVE TESTED DRUGS THAT PEOPLE
HAVE REPORTED IN OUR HANDS,
DIFFICULT TO INTERPRET. SO IF
WE CAN COME UP WITH IDEAS FOR
THE COMMUNITY, THAT WOULD BE
GREAT. NOT EVERYBODY CAN DO
EVERY ASSAY BUT MAYBE ASK PEOPLE
TO DO HANDFUL OF THINGS AND THAT
WAY WE CAN BE MORE SYSTEMATIC IN
INTERPRETING PHENOTYPES.
>> GOOD POINT SARA. WONDERING IF
SOMEWHERE ALONG THE LINES WE
HAVE DISCUSSIONS AND POSSIBLE
FOLLOW-UP WONDERING IF THERE WAS
ANY BENEFIT OF SETTING UP
STANDARDIZED REAGENTS, ACCESS TO
STANDARDIZED REAGENTS, DRUGS
OTHER CONTROL TYPE THINGS THAT
COULD BE USED ACROSS THE VARIOUS
MODELS AND ASSAYS TO NORMALIZE.
I THINK WE ALL EXPERIENCE THAT
WITH SARS COV-2, PEOPLE HAVING
SLIGHTLY DIFFERENT CHALLENGE
MATERIAL AND THINGS LIKE THAT
REALLY CHANGING OUTCOMES OR EVEN
DIFFERENT SUBTLE CELL LINES,
THINGS LIKE THAT SO WE SHOULD
PUT THAT ON OUR LIST OF THINGS
TO CAPTURE BECAUSE I THINK THERE
ARE MECHANISMS IN PLACE TO GET
IT DONE SOONER RATHER THAN IN A
PANIC THE WAY WE HAVE DONE THE
LAST COUPLE OF YEARS SOMETIMES.
>> WE ARE DOING ASSAYS LIKE
WHERE DO WE LOOK OR WHAT IS
RIGHT AND I DON'T THINK THERE IS
A RIGHT ANSWER BUT WE WANT TO BE
ABLE TO HAVE OPTIONS AND KNOW
THESE ARE WHAT IS REQUIRED IF
YOU DO WANT TO DO DRUG TESTING.
>> WE ARE GETTING CLOSE TO BREAK
TO BEFORE TURNING IT OVER I WANT
TO THANK ALL THE PANELISTS AND
SPEAKERS FOR SESSION 2, IT WAS
FANTASTIC. ALSO THANK YOU TO
PARTICIPANTS WRITING IN
QUESTIONS, IF YOU ARE FEELING
THIS DISCUSSION, VERY MUCH
APPRECIATE IT. THANK YOU SO
MUCH. OVER TO SARINE FOR
TRANSITIONING.
>> THANK YOU SO MUCH, ANN, FOR A
WONDERFUL SESSION. AND ALL THE
SPEAKERS, CERTAINLY DID LIKE ALL
YOUR TALKS AND AMAZING
DISCUSSION. SO APPRECIATE Y'ALL.
SO NEXT WE WILL HAVE A BREAK
THEN RECONVENE AT 3 O'CLOCK FOR
SESSION 3 WHICH WILL BE ON USER
REPRODUCIBLE TREATING TISSUE
MODELS FROM DRUG DISCOVERY TO
DEVELOPMENT AND THAT WILL BE
CHAIRED BY DR. ROBERT JORDAN
FROM THE BILL AND MELINDA GATES
FOUNDATION. TIME FOR BREAK NOW
AND MEET AGAIN AT 3 O'CLOCK.
THANK YOU VERY MUCH.
>>WELCOME BACK. IT IS THREE
O'CLOCK SO LET'S RESUME THE
WORKSHOP. NEXT IS SESSION 3,
GOING TO BE ON THE USE OF ROBUST
AND REPRODUCIBLE MODELS FROM
DRUG DISCOVERY TO DEVELOPMENT.
I'M SUPER EXCITED TO INTRODUCE
OUR CHAIR FOR THIS SESSION WHO
IS DR. ROBERT JORDAN, SENIOR
PROGRAM OFFICER AT THE BILL AND
MELINDA GATES FOUNDATION. PRIOR
TO JOINING THE GATES FOUNDATION
ROBERT WAS VICE PRESIDENT OF
RESEARCH AND DEVELOPMENT AT MESA
VACCINES DEVELOPING LIVE
ATTENUATED VIRUS VACCINES FOR
RSV AND SARS COV-2 AS WELL AS
OTHER RESPIRATORY VIRUS
PATHOGEN. THANK YOU, ROBERT.
PASS THE BATON FOR YOU TO CHAIR.
>> WELCOME BACK TO SESSION 3.
THIS IS HAS BEEN A GREAT MEETING
SO FAR WE HAVE HEARD INTERESTING
RESEARCH ON 3D TISSUE MODELS TO
BETTER UNDERSTAND COMPLEX
BIOLOGY OF VIRUS HOST
INTERACTIONS AND HOW THEY CAN BE
USED TO FACILITATE DRUG
DISCOVERY AND DEVELOPMENT FOR
ANTIVIRALS AND UP COVER
INTERESTING PHENOTYPES THAT ARE
ASSOCIATED WITH ANTIVIRAL
DISCOVERY THAT AREN'T READILY
AVAILABLE IN 2D SYSTEMS, THERE'S
A LOT OF IMPORTANT INFORMATION
PRESENTED SO FAR. I THE GOAL OF
THIS SESSION IS TO LOOK AT
PRACTICAL ASPECTS OF 3D TISSUE
CULTURE MODELS IN DRUG DISCOVERY
AND DEVELOPMENT. SO LET ME
INTRODUCE OUR FIRST SPEAKER
TONIGHT, OUR FIRST SPEAKER IS
DR. NICOLE K ANY KLEINSTREUER H.
FEDERAL RESOURCE FOR
ALTERNATIVES TO ANIMAL TESTING.
ATH AT THE NICEATM SHE LEADS
DOMESTIC INTERNATIONAL EFFORTS
TO DEVELOP NOVEL TESTING
ANALYSIS STRATEGIES TO PROVIDE
RAPID MECHANISTIC AND HUMAN
RELEVANCE PREDICTIONS OF
POTENTIAL ENVIRONMENTAL HAZARDS.
DR. KLINESTREUER FOCUS FOCUSES
ON THE MATHEMATICAL MODELING OF
PERTURBATIONS AND ADVERSE
OUTCOMES. THE TITLE OF HEIFER
TALK IS CONNECTING AND
SUPPORTING THE TISSUE MODELING
COMMUNITY VIA NPSCORE GLOBAL
WORKING GROUP. NICOLE, THE FLOOR
IS YOURS.
>> PERFE PERFECT KLEINSTREUER. T
YOU CAN SEE MY SCREEN IN THE
APPROPRIATE VIEW. SO YEAH, I AM
AS I LEAVE NICEATM USUALLY HOW
WE SAY IT BUT I DO ACCEPT THE
FRIENDLY CASH MACHINE NICE ATM
PRONUNCIATION AS WELL. SO THIS
IS REALLY EXCITED TO TALK TO YOU
TODAY ABOUT THIS INITIATIVE THAT
WE PUT FORWARD OVER THE LAST TWO
YEARS NOW, THIS STARTED ABOUT
SIX MONTHS INTO THE PANDEMIC AND
I HAVE TO ABSOLUTELY START WITH
A SHOUT OUT TO MY CO-CHAIR ON
THIS WORKING GROUP ANTHONY
HOLMES SO A JOINT VENTURE
BETWEEN THE UK NC 3Rs TO, A
MALL CENTER REDUCTION REFINEMENT
AND REPLACEMENT OF ANIMAL
TESTING IN ENGLAND AND SO HE AND
I GOT TOGETHER ABOUT TWO YEARS
AGO AND DECIDED IT WAS TIME FOR
US TO TRY ARE TO CREATE THIS
NETWORK TO BRING THE COMMUNITY
TOGETHER ON FOLKS USING NPS
MODELS TO STUDY INFECTIOUS
DISEASE SPECIFICALLY COVID TO
PROVIDE A REALLY RESPONSIVE
EFFORT TO THE PANDEMIC MOTIVATED
BY THE FACT THAT THERE ARE NO
TRULY GOOD ANIMAL MODELS FOR
COVID DISEASE PRESENTATION AND
WE REALLY SAW IT AS AN
OPPORTUNITY TO HIGHLIGHT AND
INVESTIGATE THE UTILITY OF HUMAN
BIOLOGICALLY BASED
MICROPHYSIOLOGICAL SYSTEMS AND
PROVIDING INSIGHT INTO DISEASE
MECHANISMS AND TEST THERAPEUTICS
AND COORDINATE ACTIVITIES AROUND
MPS AND COVID-19 TO MAXIMIZE
HUMAN HEALTH AND ANIMAL WELFARE
IMPACTS OF THESE PLATFORMS. SO
FIRSTLY START WITH A LITTLE
BACKGROUND ABOUT THE WORKING
GROUP SO THE FOUNDING MEMBERS
ARE ON THE SCREEN HERE SO MYSELF
AND ANTHONY AND EXCELLENT
SUPPORT FROM NCATS NOT A
SURPRISE GIVEN THEIR TISSUE CHIP
CONSORTIUM ACTIVITIES. SO THEY
REALLY HELPED PUT US IN TOUCH
WITH ALL THE FOLKS THAT WERE
ALREADY GRANTEES UNDER THE NCATS
TISSUE CHIPS CONSORTIUM AND
RECEIVED SUPPLEMENTAL FUNDING
FOR DOING COVID-19 RESEARCH IN
THEIR PLATFORMS. AND THEN OF
COURSE WE ABSOLUTELY NEEDED TO
ENGAGE WITH NIAID AND MARK
WILLIAMS FROM D MID HAS BEEN A
FANTASTIC RESOURCE. THE U.S.
ARMY IS DOING REALLY EXCELLENT
WORK IN THE TISSUE CHIP SPACE
AND TYLER AND KYLE HAVE PROVIDED
QUITE LOT OF EXPERTISE AND
LEADERSHIP OF FOUNDING MEMBERS
OF THE THIS GROUP AS WELL. SO
WHO ARE OUR MEMBERSHIP? ONE OF
THE MAJOR OBJECTIVES WAS TO TRY
AND ENGAGE REGULATORY DECISION
MAKERS AND GOVERNMENT AGENCY AS
MUCH AS POSSIBLE TO START TO
UNDERSTAND WHAT ARE THE NEEDS IN
THIS TYPE OF SPACE, SO IN
INFECTIOUS DISEASE RESEARCH AND
LOOKING AT NEW THERAPEUTICS IN
UNDERSTANDING SAFETY AND
EFFICACY FROM A HUMAN BIOLOGY
BASED PERSPECTIVE AND GETTING
THAT FEEDBACK AND THAT DIALOGUE
WITH REGULATORS SO THAT THE MPS
COMMUNITY CAN BE BUILDING MODELS
AND DOING EXPERIMENTS THAT ARE
GOING TO PROVIDE INFORMATION
THAT IS VALUABLE TO REGULATORY
AUTHORITIES. SO WE HAVE
REPRESENTATIVES FROM DIVISIONS
BRANCHES OR CENTERS OF ALL OF
THE AGENCIES YOU SEE ON THE
SCREEN HERE OBVIOUSLY WE ARE
PRETTY U.S. CENTRIC BUT WE HAVE
HAD SOME REALLY GOOD INTERACTION
WITH EUROPEAN REGULATORS AS
WELL. WE HAVE QUITE A FEW
BIOTECH COMPANIES REPRESENTED,
SO THESE ARE SOME OF THESE ARE
SPIN OFFS TO SOME OF THESE
LARGER COMPANIES THAT HAVE
DEVELOPED MPS MODELS IN THEIR
OWN RIGHT AND COVERING A WIDE
VARIETY OF ORGANS. THEN WE HAVE
PARTICIPATION FROM LARGE
PHARMACEUTICAL COMPANIES THAT
ARE QUITE INTERESTED IN USING
THESE MPS PLATFORMS TO TEST
THINGS LIKE NEW THERAPEUTICS
THAT ARE IN THEIR PIPELINE. THEN
THE LARGEST CONTINGENT WE HAVE
REPRESENTATION FROM IS THE
ACADEMIC RESEARCHERS BECAUSE A
LOT OF THE CUTTING EDGE SCIENCE
AND LOT OF REALLY EXCITING WORK
IN MPS IS GOING ON IN ACADEMIC
LAB SO H IS JUST A SLIDE ONE OF
TWO SO WE HAVE GROUPS FROM ALL
OF THESE DIFFERENT INSTITUTIONS
REPRESENTED ON THE MPS CORE
WORKING GROUP. SOME -- QUITE FEW
MEMBERS HAVE PRESENTED IN THIS
WORKSHOP OVER LAST DAY AND A
HALF. THEN THIS IS OUR SECOND
SLIDE SHOWING AFFILIATIONS.
AGAIN REALLY INTERNATIONAL
PRESENCE SO NOT JUST EU AND U.S.
AND NORTH AMERICA AND UK BUT
ALSO WE HAVE REPRESENTATION FROM
ASIA, LATIN AMERICA AS WELL.
WHAT WERE THE OBJECTIVES
INFORMING THIS WORKING GROUP?
FIRST AND FOREMOST IT WAS TO
UNITE A SAFE SPACE FOR
COMMUNICATIONS AND ENGAGEMENT
AMONG RESEARCHERS TRYING TO
UNDERSTAND COVID DISEASE
MECHANISMS, TRYING TO COME UP
WITH PLATFORMSNA ARE MUCH MORE
HUMAN RELEVANT TO UNDERSTAND THE
PATHOPHYSIOLOGY AND TEST NEW
THERAPEUTICS TO RAISE THAT
AWARENESS OF MPS TECHNOLOGIES
AND UTILITY WITH RESPECT TO
COVID-19 BUT ALSO WITH RESPECT
TO FUTURE INFECTIOUS DISEASE
RESEARCH. TO TRY TO FACILITATE
CONNECTIONS BETWEEN FOLKS
DEVELOPING TECHNOLOGY AND END
USERS OF TECHNOLOGIES FROM A
PHARMACEUTICAL DRUG DEVELOPMENT
STANDPOINT OR FROM A SAFETY
ASSURANCE STANDPOINT REGULATORY
DECISION MAKING STAND POINT. I
TOUCHED THIS BUT ONE IMPORTANT
ASPECT IS TO HAVE DIALOGUE WITH
REGULAR REGULATORY AUTHORITY TO
UNDERSTAND NEEDS DECISION
CONTEXT, AND HOW MODELS ARE
CHARACTERIZED IN TERMS OF
PERFORMANCE AND READINESS
CRITERIAND USED TO SUPPORT
THINGS LIKE IND APPLICATIONS OR
DEMONSTRATIONS OF EFFICACY AND
SAFETY THAT MIGHT BE FIT FOR
REGULATORY CONTEXT AND USE.
ALSO TO MAKE CONNECTIONS AND
HAVE PEOPLE HAVE ACCESS TO
PRE-CLINICAL DATA CLINICAL
TRIALS DATA AND ASSESS WHAT IS
THE VALUE OF INFORMATION GAINED
FROM MPS MODEL AND HOW THAT
COMPARES TO THE EXISTING GOLD
STANDARD AND ARE WE GETTING MORE
INSIGHT TO THE HUMAN BIOLOGY AND
THE HUMAN RESPONSE TO DISEASE
AND THERAPEUTICS IN SUPPORT OF
OBJECTIVES ON ANIMAL WELFARE AND
REDUCING REPLACING REFINING
ANIMAL TESTING. THREE Rs,
THERE HAVE BEEN A HUGE NUMBER OF
PUBLICATIONS THAT COME OUT IN
THIS FIELD OVER THE LAST TWO
YEARS. THE VERY END AT THE TOP I
WILL HIGHLIGHT OUR WEBSITE WHERE
YOU CAN FIND MORE INFORMATION
ABOUT THIS WORKING GROUP AND
ALSO FIND A SURVEY OF THE
LITERATURE SPECIFICALLY THE
LITERATURE ON MPS AND COVID
RESEARCH, CURRENT AS OF END OF
LAST YEAR. SO I CAN'T HIGHLIGHT
THE LARGE, LARGE NUMBER OF
PUBLICATIONS THAT HAVE COME OUT
IN THIS AREA OR FROM GROUP
MEMBERS SPECIFICALLY BUT I WANT
TO HIGHLIGHT A COUPLE OF
PUBLICATIONS ONE WAS A FEATURE
ARTICLE THAT WAS PUBLISHED IN
DRUG DISCOVERY THAT ANTHONY AND
I CO-AUTHORED TO TALK ABOUT THE
INTROINTRODUCE THE CORE WORKING
GROUP TO THE SCIENTIFIC
COMMUNITY BUT ALSO TALK ABOUT
THE CHALLENGES THAT ARE
AFFECTING THE UTILITY AND THE
ADOPTION OF MPS FOR SARS COV-2
RESEARCH AND DRUG DEVELOPMENT SO
WE SENT A SURVEY TO THE MPS FOR
WORKING GROUP MEMBERS AND ASKED
THEM TO SELECT THEIR TOP THREE
SCIENTIFIC PRACTICAL CHALLENGES
OR BARRIERS THAT ARE AFFECTING
THE USE OF MPS FOR SARS COV-2
AND COVID RESEARCH. SO YOU CAN
SEE SOME OF THE ANSWERS WE GOT
BACK ON THE SLIDE HERE. SO THE
TOP CHALLENGE FROM SCIENTIFIC
STANDPOINT WAS INCORPORATING
HOST IMMUNE AND CORE IMMUNE
COMPONENTS FOLLOWED BY THINGS
LIKE PERFORMANCE CRITERIA, THEN
ACCESS TO TISSUES, AND ACCESS TO
TISSUE AND CELL SOURCES. BOTH
NORMAL HUMAN TISSUES DISEASED
HUMAN TISSUES AND THERE WERE
SOME OTHER CHALLENGES THAT WERE
TECHNICAL IN NATURE LIKE PBMS
DRUG BINDING AND OTHERS THAT
WERE MORE I THINK PRACTICAL IN
NATURE, DARE I SAY PHILOSOPHICAL
IN TERMS OF LACK OF CONFIDENCE
IN MODELS OR WARENESS OF THE
MODEL AND CONTEXT OF USE. ON THE
PRACTICAL CHALLENGES SIDE THERE
WAS AGAIN ISSUES AROUND RESOURCE
ALLOTMENT SO FUNDING BY FAR, THE
NUMBER ONE ISSUE BUT ALSO ACCESS
TO BSL 3 FACILITIES, THE ABILITY
TO WORK WITH LIVE VIRUS REQUIRES
YOU HAVE THIS BIOSAFETY LEVEL OF
THREE OR FOUR AND THOSE ARE NOT
ONLY CHALLENGING TO FIND BUT
ALSO CHALLENGING TO WORK IN EVEN
ONCE YOU HAVE ACCESS TO THEM. SO
SOME OF THESE CHALLENGES THAT WE
ACCUMULATED FROM THESE SURVEY OF
OUR MEMBERSHIP ARE BEING
ADDRESSED BY INTERACTION AND
DISCUSSION AMONG OUR MEMBERSHIP.
AND BY SPECIFIC PROJECTS WE
SUPPORT THROUGH THIS WORK GROUP
I WILL TALK ABOUT A LITTLE BIT
LATER. THE OTHER PUBLICATION I
WANT TO HIGHLIGHT BUT WON'T
SPEND MUCH TIME ON BECAUSE YOU
ARE HEAR MORE ABOUT THIS WORK
LATER IN THIS SESSION FROM
THOMAS HARTUNG BUT THOUGHT IT
WAS A NICE PUBLICATION THAT WAS
AN ILLUSTRATION HOW MPS ARE
ABSOLUTELY CRITICAL RESOURCE FOR
COVID-19 RESEARCH AND IN THIS
CASE PARTNER WITH THE JOHNS
HOPKINS GROUP AND NIAID
RESEARCHERS TO DO SURVEY OF THE
LITERATURE OF ORIGINAL RESEARCH
ON SARS COV-2 NEUROTROPISM.
USING BRAIN ORGANOID MODELS. I
WON'T SPOIL WHAT THOMAS IS GOING
TO TELL YOU BUT WE ALSO LOOKED
AT SYMPTOMS OF COVID-19 PATIENTS
ASSOCIATED WITH CENTRAL NERVOUS
SYSTEM AND THEN FROM THAT
SUGGESTED WHAT THE KEY
MECHANISMS OF SO CALLED
NEUROCOVID MIGHT BE IN TERMS OF
HOW INFECTION IN THE BRAIN MIGHT
LEAD TO ALTER NEUROCHEMICAL
LANDSCAPE, ALTER BEHAVIOR, OTHER
NEUROLOGICAL END POINTS.
SO THE PROJECTS THAT ARE FUNDED
DIRECTLY UNDER THE MPS CORE
WORKING GROUP ARE TWOFOLD. ONE
IS COLLABORATION BETWEEN MY
GROUP DOD AND NIAID. THAT IS
ESTABLISHING ORGAN ON CHIP
TECHNOLOGY AT THE NIAID
INTEGRATED RESEARCH FACILITY
WHICH HAS BEEN IN ANIMAL SHOPS A
VERY STRONG NON-HUMAN PRIMATE
PROGRAM AND OTHER ANIMAL MODELS,
BUT NOW FOR THE FIRST TIME THEY
HAVE MPS IN HOUSE AND THEY ARE
CONDUCTING THIS INITIAL PROOF OF
CONCEPT STUDY THAT WILL ALLOW
SIDE BY SIDE EVALUATION OF LOW
AND HIGH COMPLEXITY MODELS ANDLY
TALK ABOUT THAT IN MORE DETAIL
THEN THE MPS DATABASE IS A
WEB-BASED RESOURCE SUPPORTED BY
NCATS TO ACCELERATE DEVELOPMENT
AND APPLICATION OF MPS FOR BOTH
BASIC RESEARCH AND DRUG
DEVELOPMENT SAFETY AND EFFICACY.
SO WE HAVE PARTNERED WITH THOSE
DEVELOPERS TO PROVIDE ADDITIONAL
SUPPLEMENTARY FUNDING TO EXPAND
THE PLATFORM AND INCLUDE
COVID-19 DISEASE PORTAL. I WILL
SHOW YOU WHAT THAT LOOKS LIKE IN
A SECOND. SO FIRST THE NIAID
COLLABORATION. ESTABLISHING THE
LUNG CHIP TECHNOLOGY AT THE IRF
AND THEN DOING SOME EXPERIMENTS
IN BOTH BSL 2 AND 4 ENVIRONMENTS
STARTING WITH SOME SYNTHETIC
HUMAN DERIVED ANTIBODIES THAT
DEMONSTRATED EFFICACY POTENCY
AGAINST SARS COV-2 ENTERING
HUMAN CLINICAL TRIALS IN EUROPE
AND CANADA. BEING ABLE TO DO
THIS SIDE BY SIDE EVALUATION OF
LOW AND HIGH COMPLEXITY MODELS
BY COMPARING TYPICAL IN VITRO
CELL CULTURE LIKE VIRO CELLS
WITH MPS MODELS WITH ANIMAL
MODELS LIKE THE GOLDMAN HAMSTER
WITH HUMAN DATA FROM CLINICAL
TRIALS, TO UNDERSTAND WHAT IS
VALUE OF INFORMATION GAINED FROM
USING THESE MORE HUMAN RELEVANT
MPS PLATFORMS. THEN THE MPS KB
IS NOW CALLED BIOSTATISTICS
ANALYTIC PLATFORM, PART OF THEIR
GRANT FROM NCATS OR FUNDING FROM
NCATS INCLUDES COMMERCIALIZATION
PHASE SO MY UNDERSTANDING IS
THAT IT WILL CONTINUE TO BE FREE
OF ACCESS TO CERTAIN CUSTOMERS
LIKE ACADEMIC AND GOVERNMENT
INSTITUTIONS BUT FOR THINGS LIKE
LARGE PHARMACEUTICAL COMPANIES
THAT ARE DEVELOPING COMMERCIAL
VERSION THAT WILL ALLOW YOU TO
DO WORK CONFIDENTIALLY BEHIND A
FIREWALL AND IT IS PHENOMENAL
RESOURCE, IF YOU HAVEN'T CHECKED
IT OUT I ENCOURAGE YOU TO DO SO.
THERE ARE NOT ONLY IS THERE
INFORMATION ABOUT DIFFERENT MPS
MODELS AND UTILITY AND DIFFERENT
PROJECTS AND EXPERIMENTS
ONGOING, THERE IS ALSO
COMPUTATIONAL TOOLS AND WORK
FLOWS YOU CAN APPLY TO YOUR OWN
DATA SO YOU CAN LOOK AT THINGS
LIKE REPRODUCIBILITY ANALYSES,
RUN PHYSIOLOGICALLY BASED
PHARMACOKINETIC MODELS TO
UNDERSTAND ADME AND TOX AND LOOK
AT COMPARING SIDE BY SIDE WITH
OTHER EXPERIMENTS TARGETING
BIOLOGY OR MECHANISMS INTERESTED
IN QUERYING. THERE IS AGAIN
DIFFERENT LEVELS OF SECURITY SO
YOU CAN LOAD DATA AND
INFORMATION ABOUT MODEL SYSTEM
AND KEEP COMPLETELY CONFIDENTIAL
TO YOURSELF OR YOUR GROUP OR
SELECT GROUP OF COLLABORATORS OR
MAKE IT PUBLICLY AVAILABLE. SO
IT IS A TREMENDOUS RESOURCE. WE
FUNDED THE DEVELOPMENT OF
COVID-19 DISEASE WITHIN THE
BIOSTATISTICS ANALYTICS
PLATFORM, THAT HAS LOTS OF
INFORMATION ON IT SO LINKS TO
RESULTS OF DIFFERENT LITERATURE
SEARCHES, LINKS TO EXPERIMENTAL
MODELS THAT ARE ALREADY BEING
USED FOR COVID-19 DISEASE
RESEARCH SPANNING THAT WHOLE
RANGE FROM IN VITRO TO IN VIVO,
RESOURCES THAT IN TERMS OF
ACTUAL SUPPLIES YOU NEED TO
PURCHASE, DIRECT LINKS TO
INVENTORY TORRS, COMPARING TO
COMPUTATIONAL MODEL OUTPUTS,
PRE-CLINICAL DATA, CLINICAL
DATA, ET CETERA. OUR MEMBERS
HAVE ACCESS TO A PRIVATE PORTAL
THAT ALLOWS YOU TO IDENTIFY
OTHER RESEARCHERS THAT ARE
WORKING IN THE SAME AREA SO THE
SAME TISSUE TYPE OF INTEREST OR
SAME PLATFORM OR MAYBE HAVE BSL
3 OR 4 FACILITIES AVAILABLE AND
THIS IS A PRIVATE PORTAL ONLY
ACCESSIBLE TO WORK GROUP
MEMBERS. THE MOST RECENT THING
WE HAVE DONE IS PROVIDED
ADDITIONAL FUNDING ON TOP OF
THAT TO CREATE A PORTAL FOR
INDUCED PLURIPOTENT STEM CELLS.
THAT WAS ONE CHALLENGE
IDENTIFIED IN OUR MEMBER SURVEY,
ACCESS TO IPSCs AND ACCESS TO
INFORMATION ABOUT IPSCs SO
WELL CHARACTERIZED CELLS. HERE
WE HAVE INFORMATION AND LINKS ON
HOW TO OBTAIN CELLS, WHAT ARE
CHARACTERISTICS, HOW TO USE THEM
PROTOCOLS ET CETERA, SO
CURRENTLY WITHIN THE IPSC PORTAL
FOR EACH INSTANCE WE HAVE THE
DIFFERENT CHARACTERISTICS LIKE
THE DIFFERENTIATION, LEVEL AND
STATUS AND PHENOTYPES, THE
PATIENT PROFILES WITH CELLS
DERIVED FROM, SOURCE OF CELLS
PROTOCOLLINGS AND ESTABLISHED
APPLICATION SO WE HAVE NUMBER OF
RESOURCES AVAILABLE THIS IS THE
MAIN WEBSITE ON THE NICE ATM
HOME PAGE TO ACCESS WE HAD
SATELLITE TISSUE SHIP CONSORTIUM
MEETING AND FULL RECORDING IS
AVAILABLE SO REALLY TREMENDOUS
PRESENTATIONS, FROM OUR MEMBERS.
WE ALSO HAVE INTERNAL WEBINARS
NOT MADE PUBLICLY AVAILABLE BUT
INTENDED TO CONNECT RESEARCHERS
TOGETHER AND TROUBLESHOOT
HANDSOME APPROXIMATE FOLKS MAYBE
ENCOUNTERING, ALSO A
DISTRIBUTION LIST FOR THAT
PURPOSE. MY GROUP IS WE HAVE
EXPERTS IN VALIDATION OF
ALTERNATIVES AND WE HAVE OTHER
MEMBERS EXPERT IN THAT SPACE, SO
HAPPY TO HELP PROVIDE STUDY
DESIGN SUPPORT, DISTRIBUTE
INFORMATION ON FUNDING
MECHANISMS AND TRYING TO ENGAGE
AS MUCH AS POSSIBLE WITH
REGULATORY DECISION MAKERS. ONE
REALLY EXCITING ACTIVITY WE
PARTICIPATED IN AT THE END OF
LAST YEAR THROUGH SIMON FUNNELL
AND WHO WAS TRYING TO ENGAGE TO
IDENTIFY MPS RESEARCHERS THAT
MIGHT BE READY TO HIT THE GROUND
RUNNING TO START DOING
EXPERIMENTS ON VARIANTS OF
CONCERN. PROVIDING THIS AGILE
RESPONSE NETWORK WHERE THEY
COULD GO THROUGH WHO
DISTRIBUTION NETWORK AND GET THE
VARIANTS OF CONCERN AND
IMMEDIATELY TEST THEM AS THEY
ARISE AND EMERGE. THEN THEY
ALREADY MENTIONED MPS DB
BIOSTATISTICS PORTAL IS
FANTASTIC AND ON THEIR WEBSITE
THEY HAVE A NUMBER OF TRAINING
RESOURCES AVAILABLE THAT
ENCOURAGE YOU TO CHECK OUT. SO
I WILL STOP THERE, THANK YOU FOR
YOUR TIME AND SAY IF YOU ARE NOT
A MEMBER OF THE MPS CORE WORK
GROUP ALWAYSOR AND WOULD LIKE TO
BE YOU DON'T HAVE TO ACTIVELY DO
COVID OR SARS COV-2 RESEARCH IN
MPS TO BE MEMBER, YOU CAN SIMPLY
BE DOING MPS RESEARCH OR JUST DO
COVID RESEARCH OR JUST BE
INTERESTED IN GETTING INTO THIS
SPACE. HAPPY TO HAVE YOU PLEASE
REACH OUT. THANKS.
>> THANK YOU VERY MUCH, NICOLE,
WONDERFUL TALK. REALLY
INTERESTING INFORMATION AND SURE
THERE WILL BE PLENTY OF
QUESTIONS AT THE END OF THE
SESSION. OUR NEXT SPEAKER IS EVI
STRUBLE WHO IS A RESEARCH
PHARMACOLOGIST DERIVATIVES
BRANCH DIVISION OF PLASMA
PROTEIN THERAPEUTICS OFFICE OF
TISSUE CENTER FOR EVALUATION
BIOLOGICS AND RESEARCH AT THE
U.S. FOOD AND DRUG
ADMINISTRATION. Ph.D. IN
BIOPHYSICS FROM JOHNS HOPKINS
UNIVERSITY POST-DOCTORAL
TRAINING AT THE NATIONAL
INSTITUTE OF STANDARDS AND
TECHNOLOGY. SHE IS RESEARCH
PHARMACOLOGIST AT CBER FDA SHE
PERFORMS REGULATORY
APPLICATIONS, AND ADDITIONALLY
RELATED RESEARCH. AS A RESEARCH
SCIENTIST DR. STRUBLE EXAMINES
ANIMAL SAFETY DRUGS DURING
PREGNANPREGNANCY. RECENT WORK ON
THERAPIES FOR ZEKA VIRAL
INFECTION. THE TITLE OF HER TALK
TODAY IS MICROPHYSIOLOGICAL
SYSTEMS FOR EVALUATING IGG
ANTIVIRALS DURING PREGNANCY.
EVI.
>> THANK YOU. THANK YOU FOR THE
INVITATION TO TALK. LET ME SHARE
MY SCREEN. DO YOU SEE MY SLIDES
SM
>> YOU NEED TO SHIFT TO SO WE
CAN SEE -- YOU ARE IN PRESENTER
MODE SO WE SEE THE PRESENTER
MODE.
>> UNDERSTAND. OKAY.
>> I THINK YOU CAN SWAP ON THAT
BUT --
>> IF YOU GO BACK TO SHARE
POWERPOINT AGAIN, YOU CAN DO IT
FROM THERE. CLICK NEW SHARE,
FROM THERE
>> IS IT BETTER?
>> NO, GO BACK TO POWERPOINT
AGAIN. YOUR INTERFACE IS
DIFFERENDIFFERENT. DOWN AT THE M
NEXT TO THREE DOTS, WHAT THAT
ONE?
>> HIDE PRESENTER VIEW?
>> DON'T DO THAT. NEXT TO LEFT
OVER. WHAT IS THAT ONE?
>> THAT IS SUBTITLE.
>> INITIALLY IS ON TOP, THE
SECOND --
>> USUALLY ON TOP
>> SO PROBABLY HIDDEN BEHIND
THAT BAR.
>> STOP SHARING AND I WILL MOVE
IT TO GET PRESENTER SCREEN.
>> SORRY I CAN'T SEE WHAT THE
BUTTON IS IS. ARE YOU ON A MAC?
>> NO. THE THE WEB INTERFACE OF
ZOOM? TH SO SHARE SCREEN AGAIN.
NEED TO GO BACK OVER -- OKAY
FROM HERE WE JUST NEED TO
>> IF YOU HOVER ON TOP RIGHT
NEXT TO SHOW TASK BAR HOVER TO
THE RIGHT USUALLY UNDER THAT
BAR. ONE THING WE CAN ALSO DO
ABIGAIL, DO YOU HAVE SLIDES SO
PERHAPS YOU CAN ADVANCE?
>> SHE DOESN'T. INVOLVE TO HAVE
IN PRESENTER, I CAN HAVE IT IN
REGULAR VIEW.
>> PRESENTATION VIEW IS WHAT IT
IS FOR NORMALLY CALLED IN POWER
POINT. I'M SORRY ANY ATTENDEES.
>> THERE WE GO.
>> I'M SORRY, ABOUT THIS
TECHNICAL DIFFICULTIES. I WAS
ASKED TO TALK ABOUT SOME OF OUR
WORK WE HAVE BEEN PERFORMING
TOWARDS DEVELOPING A
MICROPHYSIOLOGICAL SYSTEM, ONE
GROUP WE HAVE BEEN DUB IN
COLLABORATION WITH CENTER FOR
TOX LOGICAL RESEARCH IS MY
COLLABORATOR AND THIS IS A NEW
WORD DOING THIS ABOUT YEAR AND A
HALF OR SO IN ADDITION TO TIE
INTO THAT TALK ABOUT THE WAY WE
THINK AT THE FDA IN OUR BRANCH
AND OUR DIVISION ABOUT UTILITY
OF SUCH MOSHING PHYSIOLOGICAL
EXPERIMENTS WHEN WE REVIEW
ANIMAL DATA. I'M SORRY,
PRE-CLINICAL DATA FOR MAKING
REGULATORY DECISIONS. SO
DISCLAIMER THOUGH MY SLIDES HAVE
BEEN CLEAR WHATEVER I SAID
VERBALLY IS ANY BEST JUDGMENT
AND DOES NOT BIND THE FDA. SO WE
ARE TALKING ANTIVIRALS, IT IS
NORMAL TALK ABOUT INFECTIONS AND
INFECTIONS ARE SOMETHING THAT
CAN HAPPEN DURING PREGNANCY AND
PREGNANT WOMEN ARE VERY
SUSCEPTIBLE TO INFECTION THEY
CAN RESULT IN MATERNAL AND FETAL
TOXICITY HERE I LIST SOME
INFECTIONS THAT ARE OF
RELEVANCE. especially the
health of the baby. The fetal
and developmental issues can
occur via direct infection of
babies but also through
infection of the placenta and
the problems that arise such as
decrease nutrient oxygen
transport presence of reservoir
for infection and et cetera.
The -- what so the antibody
treatment and antibody
therapeutics can be used during
pregnancy for to bomb cat such
infections and here is a table
of some examples of antibodies
that are being used, or can be
used or are being studied for
use, during pregnancy for viral
infections. I subdivided the
table here in two categories.
One antibodies that are
indicated for pregnancy. For
indications such as further
prevention of transmission of
infection, that includes
cytomegalovirus, hepatitis B
HIV, there is an example of
antibody cocktail that is
studied for the prevention of
Zika neurological damage in
macaques. Rhesus macaques but
in addition to this group there
is another subgroup that
includes antibodies not approved
for pregnancy per se but there
include treatment for eBola and
SARS COV-2 and for such
antibodies the prescribing
information as well as the fact
sheet or EUA clearly state that
the antibody treatment should
not be held to pregnancy women
-- pregnant women due to if risk
benefit ratio is -- these
therapies have the same time
line and drug development as
other drugs. I'm showing this is
the linear array, we are forward
moving type of development which
we very well know is not such.
So for example, the clinical
trials, can be ongoing in
healthy volunteers while
pre-clinical studies in for
example to support use
susceptible population can be
ongoing. Because yes talking
microphysiological systems, in
vitro systems that can be used
in this part of the
developmental process, in the
pre-clinical non-clinical
assessment. The purpose of this
pre-clinical assessment is of
course to provide data to
support initiation of the
clinical trials. This type of
data falls into two buckets.
One is to provide evidence of
activity in targeting the viral
disease. And by definition these
studies are performed with in
the presence of viral agent via
live virus or suitable
surrogate. These studies are
often we call them proof of
concept or Pharmacodynamic
studies not only aimed to show
the activity but most
importantly they need 'ally dose
to biologic activity. So they
are aimed to identify those
activity -- dose activity
relationship. On the other hand,
the other bucket of studies is
study that show evidence for
safety, these are toxicology
studies and most often not
always these studies are
performed in healthy animals. In
general there's no in vitro
alternatives or substitutions
for these general toxicity
studies, however, in vitro
studies are.
PROKED FOR SPECIFIC ASPECT OF
TOXICITY AND SAFETY AND I WILL
HOPEFULLY HIGHLIGHT ONE OR TWO
OF THOSE IN MY SUBSEQUENT
SLIDES. BUT I WOULD LIKE TO
MENTION HERE THAT WE ENCOURAGE
USE OF CLEAR PRINCIPLES WHEN
DESIGNING THESE TOXICITY
STUDIES. PROOF OF CONCEPT
PHARMACO DYNAMIC STUDIES
PHARMACODYNAMIC STUDIES ARE
CALLED POTENCY ASSAYS, THEY
PROVIDE A QUANTITATIVE MEASURE
OF THE ACTIVITY AS LINKED AND
DEPENDENT ON THE MECHANISM OF
ACTION. ICH QCB LAYS OUT THEY
ARE -- THESE TYPES OF ASSAYS CAN
BE IN VIVO INRY VITRO OR
BIOASSAYS. FOR ANTIVIRALS
INCLUDING ANTIVIRAL ANTIBODIES,
IS NEUTRALIZATION ASSAY. THAT IS
MOST OFTEN IN VITRO ASSAY,
SOMETIMES WE SEE BY AND LARGE
THESE ASSAYS ARE IN VITRO AND
THESE SCHEMATIC ON THE RIGHT
THAT/MY COLLEAGUE OFFICE OF
VACCINES, DR. WISE, YOU CAN SEE
THAT THIS ASSAY CAN BE PERFORMED
WITH WHOLE VIRUS IN THE CASE OF
SARS COV THEY HAVE TO BE
PERFORMED UNDER BSL 3
CONDITIONS, BUT ALSO IT IS IN
OTHER CASES TO OVERCOME THIS
CHALLENGE SO THE VIRUS CAN BE
USED. AND -- (INAUDIBLE) CAN BE
OF DIFFERENT KINDS BUT THEY HAVE
THE ADVANTAGE WHEN VIRUS IS
PSEUDO VIRUS IS USED, THIS
ASSAYS COULD BE PERFORMED UNDER
BSL 2 CONDITIONS. HOWEVER, WE
OFTEN REQUEST AND REQUIRE THAT
THE DATA FROM ASSAYS THAT ARE
PERFORMED WITH PSEUDO VIRUS BE
CORRELATED TO THE DATA FROM
ASSAYS PERFORMED WITH ACTUAL
CLINICAL ISOLATE OR INFECTIOUS
VIRUS. SO IN VITRO UTILIZATION
ASSAYS HAVE ADVANTAGES AND DRAW
BACKS AND THE ADVANTAGES ARE
OBVIOUS, LONG HISTORY, MANY
OPTIONS, PLAQUE NEUTRALIZATION
ASSAY, ET CETERA. ANOTHER
IMPORTANT POINT HERE IS THAT FOR
THIS ASSAY THEY ARE AUTOMATABLE,
THEY CAN BE USED ROUTINELY FOR
EXAMPLE MICRONEUTRALIZATION
ASSAY. THEY CAN DERIVE ACTIVITY
CONCENTRATION RELATIONSHIP AS I
ALREADY MENTION AND THEY ALSO DO
NOT INCLUDE USE OF ANIMALS.
HOWEVER THE DRAWING BACKS ALSO
ARE CLEAR, SIMPLIFIED MODELS,
THEY ARE A SNAP SHOT IN TIME, WE
DON'T USUALLY GET KINETIC
INFORMATION WITH THESE ASSAYS.
ALSO WE DON'T USUALLY GET ORGAN
CONCENTRATION INFORMATION. WE
GET CELL LEVEL BUT ORGAN
CONCENTRATION. AND CONVERTING
ACTIVE CONCENTRATION, IC 19s
TO DOSE CAN BE CHALLENGE
ESPECIALLY FOR MORE COMPLICATED
SITUATIONS SUCH AS PREGNANCY.
AND WE VIEW MICROPHYSIOLOGICAL
SYSTEMS AS A BRIDGE BETWEEN IN
VITRO AND IN VIVO, NOTHING NEW
HERE BUT ALSO THE ADVANTAGES OF
MICROPHYSIOLOGICAL SYSTEMS ARE
CLEAR VERSUS SINGLE CELL
NEUTRALIZATION ASSAY. SO WE
DECIDED, MYSELF AND COLLABORATOR
DECIDED TO FOCUS OUR EFFORT
DEVELOPING A PLACENTA IPS FOR
SEVERAL REASONS ONE BECAUSE WE
SAW THERE IS A NEED PLACENTA IS
IMPORTANT ORGAN CRITICAL FOR
HEALTH OF THE PREGNANT MOTHER
AND FETUS. THERE IS -- IT IS
ALSO AN ORGAN NOT VERY WELL
STUDIED AND USED BECAUSE -- THAT
ACTUALLY IT IS A MISSING THAT
ACTUALLY IS AN AREA THAT NEEDS
TO BE IMPROVED. PLACENTA IS
ACCEPTABLE TO -- SUSCEPTIBLE TO
INFECTION AND DAMAGE AND WE
DECIDED TO USE ZIKA AS A CASE
STUDY BECAUSE IT WOULD ALLOW US
TO LOOK AT THE PATHOYENSIS OF
PLACENTA INFECTION IN GENERAL AS
WELL AS LOOK INTO A SPECIFIC
THEORETICAL CONCERN, WORRIED
ABOUT, WHAT IS KNOWN AS ANTIBODY
ENHANCEMENT WHICH IS SHOWN IN
VITRO SYSTEM OF ANIMAL. FINALLY
MODELING A PLACENTA IN A IN
VITRO MPS WOULD BE USEFUL TO
ALSO LOOK AT THE ANTIBODY
TRANSFER WHICH IS ANOTHER THING
UNIQUE FOR ANTIBODIES WHEN THEY
ARE USED IN PREGNANCY AND CAN
AFFECT THEIR PK AND PD FOR THE
PREGNANT WOMAN. IN DEVELOPING
OUR THREE DIMENSIONAL WE FIRST
DECIDED TO LOOK AT THE ACTUAL
WHAT ARE THE CELL TYPES AND WHAT
THAT TYPE OF STRUCTURE WE ARE
TRYING TO WE DECIDED TO MODEL
THE VILLAS. HERE IS A SCHEMATIC
OF A FIRST TRIMESTER
DIFFERENTIATED ALVEOLUS, THIS
HAS MULTIPLE CELL TYPES, IN THE
OTHER PART, THETROPE BLAST WHICH
IN CIRCLES OR ENCLOSES A
MESENCHYMEA, FETAL VESSELS ARE
ALSO EMBEDDED. GIVEN THAT THIS
STRUCTURE IS QUITE COMPLEX, WE
TOOK A STEP WISE APPROACH IN OUR
MODELING. SO WE DECIDED TO BUILD
PLACENTAL MEMBRANE BARRIERS ON A
TRANSWELL THAT CONTAIN ONLY A
MONOCULTURES THEN GO TO
CO-CULTURES AND THEN BUILD
TOWARDS A MICROPHYSIOLOGICAL
SYSTEM. THAT CONTAINS MULTIPLE
CELL TYPES. SO FOR MONOCULTURES
WE FOCUSED ON CARDIO SARCOMA
CELLS OF TRIPLE BLAST ORIGINS,
IN VIVO INJECT THREE CELL LINES
WHICH FORMS POLARIZE ISORRIZED
LAYERS IN MEMBRANE. THESE CELLS
SHOWN HERE IN THIS SCHEMATIC
SEMIPERMEABLE MEMBRANE CAN
SEPARATE IN APICAL CHAMBER WHICH
WE CAN CALL THE MATERNAL BLOOD
IF YOU WILL AS A MODEL OF THE
COMPARTMENT AND THIS CHAMBER IS
SEPARATED FROM BASAL LATERAL
CHAMBER WITH WHAT WOULD BE
EQUIVALENT OF THE FETAL
COMPARTMENT. WE FIRST BEFORE WE
EMBARKED IN MAKING THIS MEMBRANE
ON TRANSWELL WE LOOK AT THE
ABILITY OF CELL LINES TO BE
SUBSEQUENT TO ZIKA INFECTION AND
WE SHOWED HERE IS A SIMPLE
KINETIC INFECTION KINETICS OF
BIGGER CELLS INJECT THREE CELLS
MAY HAVE SAME WAY SUSCEPTIBLE TO
INFECTION AND SAW SIMILAR
KINETICS TO VIRAL CELLS. IN
ADDITION ADDING ANTI-ZIKA
FLAVIVIRUS ANTIBODIES BLOCK
INFECTION. SO THEN WE WENT TO
BUILDING THESE MEMBRANE OR
GROWING THESE BARRIERS AND
MONITOR THEIR GROWTH AND
ACTIVITY BY LOOKING AT
INTERSELECTIVE RESISTANCE AS
WELL AS THE PERMEABLE --
PERMEABILITY OF SMALL MOLECULE
TO SEE THAT THIS MEMBRANES ARE
IN FACT CONFLUENT AND HAVE LOW
PERMEABILITY. SUBSEQUENTLY WE
ADDED IN THE APICAL CHAMBER
EITHER ZIKA VIRUS WITH OR
WITHOUT ANTIBODY TARGETING ZIKA.
WE LOOKED AT THE ABILITY OF
ANTIBODY TO PASSAGE THROUGH THIS
BARRIER, INDEED WE DID SEE
ANTIBODY PASSAGE FROM CAN THE
BASAL LATERAL CHAMBERS. IN A
ADDITION SAME WAS TRUE FOR ZIKA
IT COULD PASS THROUGH BARRIERS,
ADDING ANTIBODY DECREASED THE
AMOUNT OF ANTIBODY VIRUS WE
COULD DETECT IN BASAL
COMPARTMENT SO USING THESE
BARRIERS WE DID NOT SEE
ENHANCEMENT OF INFECTION. THE
NEXT STEP BUILDING CO-CULTURE
USING DIFFERENT CELLS IN
ADDITION TOP VIVO OR THREE
TRIPLE BLAST WE ADDED
FIBROBLASTS IN THE APE AL
MEMBRANE AS WELL AS ENDOTHELIAL
CELLS HUMAN UMBILICAL VEIN
ENDOTHELIAL CELLS MANY THE
OPPOSING SIDE WE PERFORMED
SIMILAR EXPERIMENTS AS BEFORE,
FIRST AS CERTAINING THESE
MEMBRANES DID IN FACT GROW AND
CONTAIN MULTIPLE CELL TYPES, BY
HNE STAINING AND SPECIFIC
MARKERS FOR THESE CELL LINES.
THEN ADDED IGG IN APICAL
COMPARTMENT AND LOOKED FOR
PASSAGE IN BASAL COMPARTMENT AND
INDEED WE DID SEE SOME ALBEIT
SMALL AMOUNT OF IGG PASSAGE.
THIS IS MY DATA PRESENTATION WE
ARE IN THE PROCESS OF COMPLETING
AND ANALYZING THE DATA WITH
VIRUS BUT I WOULD LIKE TO SHOW
YOU WHERE WE WANT TO GO AND TO
BE IN THIS MPS SPACE. WE ARE
HOPING TO BUILD A RICH
COMPARTMENT MPS COMPARTMENT 1
REPLICATE PLACENTAL BARRIER
CONTAIN THIS MIX CELLS THEN WE
WOULD ADD IN THERE VIRUS AND IGG
OR IGM AGAIN ZIKA VIRUS,
COMPARTMENT 2 WOULD BE
EQUIVALENT OF THE FETAL
COMPARTMENT, IT WOULD CONTAIN
TISSUE SUSCEPTIBLE TO ZIKA
INFECTION SUCH AS NEUROSPHERE,
THE OUTCOMES IS TO MONITOR
PLACENTAL TOXICITY METABOLIC
FUNCTION, TRANSCYTOSIS,
INFLAMMATORY SIGNALING
MOLECULES, WE LOOK AT SIMILAR
END POINTS SUCH AS IN ADDITION
TO THAT, THE POTENTIAL FOR
INCREASE INFECTION IN THE
PRESENCE OF ANTIBODY SPECIFIC
CONCERN HAS BEEN RAISED. I WANT
TO CONCLUDE WITH A FEW THOUGHTS,
GENERAL THOUGHTS ON
MICROPHYSIOLOGICAL SYSTEMS AND
WHERE WE SEE THEM IN THE
REGULATORY SPACE AND UTILITY CAN
BE. SO AS I ALREADY MENTIONED,
WE THINK THAT THEY HAVE GREAT
POTENTIAL TO CHARACTERIZE
ACTIVITY AND MEASURE POTENCY OF
THIS ANTIVIRAL IGGs BECAUSE
THEY ARE USEFUL TO ASSESS THE
ORGAN TISSUE SPECIFIC ACTIVITY
AND WHEN COMBINED WITH
PHARMACOKINETIC OR -- AND
PHARMACODYNAMIC MODELING IT CAN
BE HELPFUL TO RELATE THOSE --
SYSTEMIC DOSE IV OR SUB Q TO
ORGAN CONCENTRATION THAT WOULD
BE POTENT AND EFFICACIOUS. HOW
FAR, REGULAR LA -- HOWEVER,
REGULATORY EXPECTATION IS THIS
AUSSIE IS PERFORMED BASED ON ONE
SCIENTIFIC PRINCIPLES AND BE
QUALIFIED AND THE QUALIFICATION
USUALLY ARE GOING TO ENTER INTO
PICTURE IN PHASE 2 AND PHASE 3
STUDIES, BECAUSE THAT IS WHERE
MORE SUSCEPTIBLE POPULATION
DISEASE WOULD BE ENTERING
CLINICAL TRIALS. THEN ANOTHER
AREA WHERE SUCH MPS COULD BE
ALSO USEFUL WOULD BE IN THE
SPECIFIC CONCERNS SUCH AS
ANTIBODY DEPENDENT ENHANCEMENT
AS WELL AS FOR EXAMPLE PLACENTAL
TRANSFER OF THERAPEUTIC
ANTIBODIES. WE SEE THIS TYPE OF
UTILITY TO BE MORE USEFUL IN THE
EARLY STAGE OF DEVELOPMENT SUCH
AS CANDIDATE SELECTION, ET
CETERA. I ALSO WANT TO IN
CLOSING TWO SUMMARY SLIDES THAT
ARE DUALLY NOT MY -- ACTUALLY
NOT MY SLIDES, I TOOK THIS FIRST
TABLE FROM A REVIEW PUBLISHED
THIS YEAR THAT CAME OUT OF A
WORKSHOP BETWEEN FDA AND
INDUSTRY ON NEW TECHNOLOGY USED
FOR REGULATORY APPLICATION. SOME
BIG IDEAS OR IMPORTANT IDEAS
THAT I UNDERLINE IN THIS TABLE
ARE SOME QUESTIONS WE NEED
ANSWERED OR ASK OURSELVES BEFORE
SUBMITTING DATA THAT HAS BEEN
DERIVED FROM NEW TECHNOLOGY. SO
ONE IMPORTANT QUESTION WOULD BE
IS THIS DATA ANSWERING
FUNDAMENTAL DRUG DEVELOPMENT
QUESTIOQUESTIONS. I HAVE UNANSWD
QUESTIONS WE ARE ANSWERING BY
USING THIS NEW TECHNOLOGY.
PREDICTIVE OF THE EFFECT THAT I
HAVE EXPECTED TO BE IN VIVO. HAS
THE VALIDITY BEEN SHOWN,
ACCURACY AND SPECIFICITY OF THE
METHOD. FINALLY, THE SLIDE WAS
TAKEN FROM A PRESENTATION DR.
JANET WEEKCOCK PRINCIPLE DEPUTY
DIRECTOR GAVE AT THE ANNUAL
MEETING OF THE COLLEGE OF
TOXICOLOGY, IT IS AN EXCELLENT
TALK, ENCOURAGE EVERYBODY TO GET
THE PRESENTATION FROM THE FDA
WEBSITE. THE POINT OF THE
SUMMARY, MAIN POINT IS FDA AND
WORLDWIDE REGULATORS WILL
INCORPORATE NEW TESTING
METHODOLOGIES. IF SOME STANDARDS
ARE MET. THE SECOND POINT IS
THAT WHEN SUCH DATA IS INTENDED
TO REPLACE AN EXISTING TEST OR
TO SUPPORT SAFETY THE BIAS IS
VERY HIGH HOWEVER IT IS VERY
IMPORTANT FIRST NEW TECHNOLOGY
TO DEFINE THE CONTEXT OF USE.
THIS MEANS HAVING A CLEAR IDEA
WHAT IS THE UNMET NEED AND
IMPROVEMENT THAT THE NEW
TECHNOLOGY IS GOING TO BRING TO
THE FLOOR. FINALLY, AFTER
CONTEXT USE IS CLEAR, THE
DEVELOPER CAN QUALIFY THE METHOD
FOR ITS USE AFTER QUALIFIED
METHOD CAN BE USED FOR OTHER
PURPOSES WITHOUT ADDITIONAL
DEMONSTRATION OF ITS
PERFORMANCE. THAT CONCLUDES MY
TALK I WOULD LIKE TO ACKNOWLEDGE
MY COLLEAGUES AT THE DIVISION OF
PLASMA PROTEIN THERAPEUTICS AS
WELL AS PEOPLE WHO PERFORM THIS
WORK, THEY ARE BOTH NOT WITH ME
ANY MORE, I'M LOOKING FOR A POST
DOC IF YOU KNOW ANYBODY OR
ANYBODY IS INTERESTED TO BRING
THIS PROJECT FORWARD PLEASE SEND
ME AN EMAIL. ACKNOWLEDGING MY
COLLABORATOR DAYTON WHO
PERFORMED STUDIES, MY OTHER
COLLABORATOR MARIA RIOS WHO IS
OUR ZIKA EXPERT, KYUNG SUNG FROM
FDA ALTERNATIVE METHODS WORKING
GROUP AND PERINATAL HEALTH
CENTER OF EXCELLENCE FOR
FUNDING. I WILL TAKE QUESTIONS.
THANK YOU.
>> THANK YOU, EVI. NICE TALK.
GET TO QUESTIONS AT THE END. I
WOULD LIKE TO INTRODUCE THE NEXT
SPEAKER DR. THOMAS HARTUNG. DR.
HARTUNG IS THE (INAUDIBLE) CHAIR
FOR EVIDENCE BASED TOXICOLOGY IN
THE DEPARTMENT OF ENVIRONMENTAL
HEALTH AND ENGINEERING JOHNS
HOPKINS BLOOMBERG SCHOOL OF
HEALTH PUBLIC HEALTH IN
BALTIMORE. DR. HARTUNG HOLDS
NUMBER OF JOINT APPOINTMENTS
INCLUDING APPOINTMENT AS
PROFESSOR PHARMACOLOGY
TOXICOLOGY UNIVERSITY OF
(INAUDIBLE) IN GERMANY, ALSO
DIRECTOR OF CENTER FOR
ALTERNATIVES TO ANIMAL TESTING
OR THE CABOT PROGRAM FOR BOTH
UNIVERSITIES. HOSTS SECRETARY OF
EVIDENCE BASED TOXICOLOGY
COLLABORATION AND MANAGES
CREATIVE COST PRACTICE, CULTURE
PRACTICE, TOXICOLOGY
DEVELOPMENTAL NEUROTOX TOXICITY
AND MICROPHYSIOLOGICAL SYSTEMS.
DR. HARTUNG'S
>> TECHNOLOGY AT HOPKINS AND
LICENSED BUT I SEE A FEW OTHERS
SO TAKE PLEASE EVERYTHING I'M
DOING WITH A GRAIN OF SALT. I
SEE BRING THE RABBIT TO THE
GROUND AND THESE WONDERFUL
TECHNOLOGIES TO ACTUAL USE.
COMING JUST FROM A MOST EXCITING
EVENT IN THE MICROPHYSIOLOGICAL
SYSTEMS LAST WEEK, WE METAPHOR
THE FIRST TIME AND DID TWO
COHORTS AND SUSIE FITZPATRICK
FROM FDA. 52 ORGANIZATIONS
TEAMED UP AND THIS IS THE BRAIN
CHILD FROM NCATS, INTO THIS REAL
WORLD SUMMIT SO THIS IS
TECHNOLOGY WHICH IS BECOMING
SCIENTIFIC DISCIPLINE OF ITS
OWN. OUR OWN CONTRIBUTION STARTS
IN 2016 WHEN WE WERE ONLY THE
THIRD OR FOURTH GROUP TO USE
BRAIN ORGANICS BECAUSE THE MINI
BRAINS AT THE TIME MASS PRODUCE
THEM SO THEY ACTUALLY COULD DO
THINGS SOUGHT SOUTH OF THESE IN
-- WHICH ARE IDENTICAL IN
CERTAIN COMPOSITION,INGS FROM
INDUCED PLURIPOTENT STEM CELLS.
YOU CAN DO THEM FROM DIFFERENT
GENETIC BACKGROUNDS FOR THIS
REASON YOU CAN ALSO MANIPULATE
EXPANSE TO NEURONAL PRECURSOR
CELLS IN 2D AND COMING AS CELL
SUSPENSION ON THIS, THEY ARE
(INDISCERNIBLE) EACH OF THESE
SIX WELL PLACE GETS US BETWEEN
600 AND 1,200 BRAIN ORGANOIDS.
SO A VERY EFFICIENT WAY OF
PRODUCING THEM. THERE ARE A LOT
OF INTERESTING FEATURES, SHOW
BIT MORE ABOUT THIS IN A SECOND.
THEY ARE SPONTANEOUS
ELECTROPHYSIOLOGICALLY ACTIVE
AND UNDERGO VARIETY OF PROCESSES
WHICH ARE TYPICAL FOR NEURAL
DEVELOPMENT. FOR EXAMPLE,
MYELINATION IS A KEY FEATURE YOU
MIGHT SEE HERE, THE LAYERS OF
OLIGODENDROCYTES AROUND AXONS,
40% AXONS ARE MYELINATED AND YOU
CAN SEE THE RESPECTEDTIVE
PUBLICATIONS AND EDITS ARTICLES
FOR THOSE WHO MIGHT BE
INTERESTED TO RETRIEVE THESE
PAPERS. THESE OLIGODENDROCYTES
WE CAN FOLLOW BECAUSE WE BROUGHT
FLOW CYTOMETRY IS WE SEE
PRE-OLIGODENDROCYTES, YOU CAN
STAIN THE BASIC PROTEIN TO MORE
MATURE OLIGODENDROCYTES SO IT IS
REALLY VERY NICE POPULATION
OFTEN ABOUT 10% OF THE CELLS WE
ARE SEEING. WE ALSO DO HAVE
OTHER CELLS, IF ASTROCYTES YOU
CAN FOLLOW WITH TIP OF YOUR
MARKERS. THE ONLY CELL THE BASE
MODEL IS LACKING IS THE
MICROGLIA BECAUSE MICROGLIA IS
INVADING CELL TYPE, IT COMES
FROM THE EGG YOLK FROM YOLK SACK
OF EMBRYO DEVELOPMENT. IT IS
PRESENT HERE. MEANTIME WE HAVE
DEVELOPED TECHNOLOGIES AND
COLLABORATION ALSO WITH THE
DEPARTMENT OF DEFENSE. FROM IPC
DERIVED MICROGROWIA WE CAN
INTEGRATE TO BRAIN ORGANOID ON
THE RESPECTIVE SLIDE. THIS IS A
VERY -- TOOL, A MODEL YOU CAN
USE FOR MODELING VARIOUS
DISEASES I LIKE TO CALL THEM
MICROPATHOPHYSIOLOGICAL MODELS.
FIRST INTERESTED IN
DEVELOPMENTAL NEUROTOXICITY,
CHEMICALS AND OTHERS LINKED TO
AUTISM DRAMATIC I INCREASE THE
OVER LAST DEGAD. ALLOWS US TO
LOOK INTO DEMYELINATION,
REMYELINATION, VARIOUS DISEASES
WHICH ARE AFFECTING THIS, WE ARE
STUDYING GENE ENVIRONMENT,
DETECTION OF DISEASES BY
BRINGING IN RISK GENES FOR
EXAMPLE, VARIOUS TYPES OF NEW
NEURODEGENERATIVE DISEASE FROM
ALZHEIMERS TO PARKINSON TO
LATERAL SCLEROSIS, WE HAVE
GRAFTED THEM IN GLIOBLASTOMA.
THIS IS THE TOPIC TODAY, WE RUN
VARIOUS INFECTION MODELS IN
THIS. THIS STARTED IN 2018 BUT
PUBLISHED ON THE FACT THAT YOU
COULD INFECT BOTH ZIKA AND
DENGUE VIRUS, WE INCLUDED ALSO
MICROGLIA, NOT YET CDNF
MICROGLIA BUT MICROGLIA LIN LINE
SHOW INFLAMMATORY RESPONSESK SEE
DAMAGE TO NEURONS DUE TO RELEASE
OF CYTOKINES AND OTHERS SO YOU
CAN MODEL INFLAMMATORY PROCESSES
HERE. MORE RECENTLY, WE
PUBLISHED ON JC VIRUS, JC VIRUS
IS A VIRUS ASSOCIATED WITH HIV
INFECTION, IT IS NOT POSSIBLE TO
PROPAGATE VIRUS IN 2D CULTURE
NORMALLY, BUT HERE IN THE HIGH
DENSITY, OF THE 3D ORGANOID,
ABOUT 1,000 TIMES DENSER THAN 2D
CULTURE THERE IS POSSIBILITY TO
DO SO. WHAT IS INTERESTING OF
THE INFECTED CELLS ON THE
PICTURES BELOW ACTUALLY NOT
NEURONS BUT THE OLIGODENDROCYTES
AND THE ASTROCYTES OF THE
SYSTEM. SO THIS IS PUBLISHED IN
FEBRUARY THIS YEAR. SO THEN THE
COVID PANDEMIC STARTED, IT
BECAME CLEAR QUICKLY THAT
DEPENDING ON THE COHORT YOU LOOK
UP TO 60% SHOW SOME TIME OF
NEUROLOGIC SYMPTOMS. VARIOUS
KINDS OF COLORFUL SPECTRUM OF
DISEASE FROM STROKE TO BRAIN
VARIOUS TYPE OF PSYCHOSIS
ENENCEPHALITIS. EVERYTHING HAS
BEEN REPORTED. ENCEPHALITIS. SO
IN MARCH OF 2020 I GAVE A
PRESENTATION AND ASKED CAN THE
BRAIN BE INFECTED. BECAUSE IT
WAS A OBVIOUS POSSIBILITY. SOME
OF THE CORONA VIRUS IS UNKNOWN
NEUROTROPIC BUT IT WAS NOT KNOWN
FOR (INAUDIBLE) AT THE TIME.
QUIZ EASIER POST THAN ANSWERED
BUT THE RICH ENVIRONMENT IN
HOPKINS ALLOWED US TO PARTNER
WITH SOME INFECTIOUS DISEASE
PEOPLE CORRY WOOD AND TEAM
AFFECTED BRAIN ORGANOIDS. THIS
WAS I THINK THE FIRST PICTURE
ACTUALLY OF HUMAN NEURON
INFECTED WITH SARS COV-2. YOU
CAN SEE THE RED DOTS EACH
REPRESENTING ONE VIRUS PARTICLE,
PARTICULARLY LARGE IN THIS CASE,
THIS GOT A LOT OF INTEREST,
FINANCIAL TIMES REPORTED ON IT
IN MAY OF 2020. 200 LAY PRESS
JOURNALS FOLLOWED AND OTHER
OUTLETS REPORTED ON THIS AND WE
WERE ACTUALLY ABLE TO FIRST
DESCRIBE IN THE PEER REVIEW
PAPER IN JUNE OF 2020, THIS IS
IN EFFECT NEUROTROPIC VIRUS.
PART OF THIS WAS SHOWING THAT
THE NECESSARY SARS 2 INHIBITOR
INHIBIT -- SARS 2 RECEPTOR FOUND
IN THE BRAIN FROM EARLY STAGES
OF NEURONAL PRECURSOR CELLS
ONWARDS. WE DID SHOW PRESENCE OF
VIRUS IN CELL -- DIFFERENT CELLS
AND ALSO REPLICATION BY PCR AND
CONFOCAL MICROSCOPY. NICOLE
REFERRED TO THIS JOINT PAPER
ABOUT A YEAR LATER WE LOOKED
INTO THE SCIENTIFIC LITERATURE
AND HOW DOES THIS STAND NOW,
THIS WAS REALLY A VERY EARLY
FINDING. YOU HAVE TO RECALL, IN
MANY APRIL OF 2020, IN THE
LITERATURE WILL IS NO ANIMAL
MODEL FOR COVID AND MANY
CLINICAL DEVELOPMENTS HAD TO GO
INTO HUMAN TRIALS IN JUNE JUST
SAFETY TESTING DATA, NOT
EFFICACY BECAUSE NO ANIMAL MODEL
WAS THERE. IT WAS AROUND SPRING
OF 2020 THIS TYPE OF MODELS ARE
DELIVERING, SHOWING VARIOUS
ORGAN CONTRIBUTIONS WHICH
ACTUALLY MAKE COVID SUCH A
DEADLY COLORFUL DISEASE. WHICH
ARE NOT REPRODUCIBLE UNTIL NOW
IN ANY ANIMAL MODEL. IF LEADING
TO LUNG INFECTION AT LEAST NOT
FROM TODAY ANY MAJOR CNS
CONTRIBUTION OF THE DISEASE. SO
JUST TO SHOW YOU A LITTLE BIT OF
THESE EXPERIMENTS WHICH HAVE
BEEN PUBLISHED THE NEWS
RELATIVELY SMALL OF OPEN VIRUS
IN THE CELL SIX HOURS EXPOSURE
THEN WE DID WASH AND WE STUDIED
THE COPY NUMBER OF THE RNA OF
VIRUS IN THE LYSATE SUPERNATANT.
AND WE DID LOOK FOR THE VIRUS
ALSO IN STAINING FOR BOTH THE
SPIKE OF THE M PROTEIN. YOU CAN
SEE THIS VIRUSES HERE, INFECTED
CELLS ARE SOME NOT VERY MANY
CELLS BUT FOUND THROUGHOUT THE
ORGANOID. YOU CAN DO CERTAIN
COUNTER STAINS. THIS IS NOW THE
SPIKE PROTEIN. IT IS NOT
CO-LOCALIZING WITH GFAP WHICH IS
THE AMOUNT OF ASTROCYTES SO NOT
THE ASTROCYTES POPULATION WHICH
IS INFECTED HERE. WE THINK SOME
TYPES OF NEURONS BEING INFECTED
I HOPE YOU CAN SEE THIS TYPE OF
PICTURES WHICH IS SHOWING A CELL
WE INTERPRET BEING SPACE LYSED
SO INFECTION PROPAGATING THE
VIRUS SET FREE. AND ABLE TO
INFECT FURTHER CELLS. IN THE
PATHOLOGY YOU CAN FIND THESE
CELLS AND YOU CAN SEE THE
CHANGES ENORMOUS AMOUNT OF
DIVERSITY CONDENSED CHROMATIN ON
THE RIGHT WHERE THE MOCK
INFECTION IS NOT SHOWING THIS
TYPE OF STRUCTURE. THIS IS THE
PCR RESULTS ON THE LEFT FOR CELL
LYSATE. YOU CAN SEE IN THESE
TOTAL 64 HOURS YOU SEE 3500 FOLD
INCREASE IN COPIES OF THE VIRUS
OVER LYSATE AND IN SUPERNATANT,
SO IT IS NOT ONLY PRODUCED IN
THE CELLS AND REPLICATING BUT
ALSO SET FREE FROM THE ORGANOID.
THIS IS AGAIN REVIEW PAPER,
WHICH NICOLE TOUCHED ON, IT IS
SHOWING MORE THAN TEN GROUPS
MEANTIME SHOWN INDEED
MICROPHYSIOLOGICAL SYSTEMS OF
THE BRAIN CAN BE INFECTED WITH
THIS VIRUS. STUDYING DIFFERENT
ASPECTS SHOWN HERE. WE SUMMARIZE
CLINICAL DATA, THEY ARE VERY
MUCH IN LINE THAT THERE IS SOME
INFECTION OF THE BRAIN BUT NOT
PERMANENT INFECTION BUT
PERSISTENT INFECTION OF THE
SYSTEM. THESE ARE SOME OF THE
PAPERS, SUMMARIZE BY OTHERS SO
MEANTIME THERE IS SOLID
LITERATURE ON NEURONAL INFECTION
BY SARS COV-2. THIS IS THE SLIDE
SHOWING NO CONSENSUS ON HOW
EFFECTS ON THE BRAIN BEING
PRODUCED, PROBABLY DIFFERENT
MECHANISMS. OBVIOUSLY IN SEVERE
COVID OF THE ENTIRE BODY WHICH
HAS AFFECT ON FUNCTIONING OF
BRAIN, THE CYTOKINE STORM, THE
PENETRATION THROUGH THE BLOOD
BRAIN BARRIER SUGGESTED BY SOME,
BUT NOT CONCLUSIVELY SHOWN.
THERE IS SOME SUGGESTION
OLFACTSRY NERVE MIGHT BE
POSSIBLE THAT RETRO GRADE
INFECTION OF BRAIN CAN TAKE
PLACE. MAINLY HYPERCORRELATION
ISCHEMIA AND VASOCONVICTION
WHICH LEADS TO REDUCED OXYGEN
WHICH IS ACTING ON THE BRAIN.
THERE HAVE BEEN CLEAR EVIDENCE
IN PATIENTS OF VIRUS IN CELLS
VIRUS IN THE CELL SPINAL FLUID,
AND ANTIBODY PRODUCTION WHICH
SEEMS TO BE FOR BRAIN INFECTION,
AND NOT IMMUNIZATION. WE ARE
CONTINUING TO WORK ON THIS
TOPICS TO IDENTIFY WHY THIS
SUBPOPULATION INFECTED WHAT
AREAS OF THE CELLS WE HAVE
PROLONGED INFECTION SCENARIO
WORKING ON BLOOD BRAIN BARRIER
ASPECT AND MOST INTERESTED ALSO
STUDY INFLAMMATORY COMPONENT AS
THIS IS A RISK FACTOR FOR
VARIOUS DISEASES AS YOU KNOW FOR
OTHER NEUROTROPIC INFECTIONS.
SEW ONE QUESTION WE ARE
INTERESTED IN IS THIS POSITIVELY
IMPACTING ON NEURAL DEVELOPMENT
ON AUTISM? BECAUSE THE BABIES OF
PANDEMIC ARE NOT YET OLD ENOUGH
TO REALLY ASSESS THIS AUTISM IS
DIAGNOSED BETWEEN 2 AND 3
SOMETIMES AS LATE AS 8 YEARS OF
PAGE. GOOD NEWS HERE IS THAT OUR
PRELIMINARY DATA KNOW THE EARLY
BRAIN DEVELOPMENT WHICH IS
VULNERABLE ASK WE BE INFECTED
VIRUS IS NOT PROPAGATING AS YOU
CAN SEE IN THE FIVE WEEK OLD
BRAIN, HERE IN BLACK, WHILE
EIGHT WEEK OLD ARE SHOWING
PROGRESSIVE INFECTION FURTHER
INCREASE VIRUS AMONG SO THIS IS
ALREADY SOME TYPE OF HOMOFUL
SCIENCE NEURAL DEVELOPMENT IS
NOT AS MUCH AND ACTUALLY THERE
IS MARKERS OF NEURAL DEVELOPMENT
BY PCR SHOW THAT THERE IS NO
MAJOR CHANGE IN COMPARTMENT OF
CELLS DUE TO INFECTION SO WE ARE
QUITE HOPEFUL THAT THIS IS NOT
STRONG IMPACTING. SO IN SUMMARY
OF THESE PARTS THIS IS A
STANDARDIZED HUMAN 3D MODEL
DEVELOPED FROM STEM CELLS, GLIAL
CELLS MYELINATION AND MICRO--
WHICH CAN BE EDIT GENETIC
BACKGROUNDS BUT INTRODUCE RISK
GENES REPAIR INCLUDE REPORTER
GENES LIKE THE ONE FOR
OLIGODENDROCYTES. INFECTION
OTHER DISEASES CAN BE MODELED
AND WE ARE VERY MUCH INTERESTED
IN GENE ENVIRONMENT INTERACTIONS
IN THE MODEL. FUNCTIONAL ASSAYS
ARE POSSIBLE NEURITE OUTGROWTH
EEG, EVEN HAVE THREE DIMENSIONAL
EEG TYPE OF ELECTROPHYSIOLOGY
AND MOST RECENTLY INTERESTED IN
IMPLEMENTING COGNITIVE FUNCTION,
ORGANOID INTELLIGENCE. I WOULD
LIKE TO SAY A FEW WORDS TIME
REMAINING. BIOLOGICAL MODELS ARE
MODEL, THEY CAN GIVE RISE TO
MANY DIFFERENT ASSAYS. AND FROM
MY UNDERSTANDING WHAT THE
DISCUSSION IS ABOUT IS ACTUALLY
ABOUT ASSAYS AND WE HAVE TO
REALLY ESTABLISH WHAT ARE LIMITS
OF THE DETECTION LIMITS OF
QUANTIFICATION, WHAT IS THE
DYNAMIC RANGE OF THESE ASSAYS WE
CAN ESTABLISH THAT DEFINE
REASONABLE BIOMARKERS POINT OF
INTEREST. THIS REQUIRES A LOT OF
THINKING MOVING MODEL TO
SOMETHING ELSE. WE ARE WORKING
AT THE MOMENT ALSO ON FREEZING
BRAIN ORGANOIDS AND THE
DIFFERENCE BETWEEN WHAT YOU SEE
HERE IS JUST ONE WAS KEPT IN
CULTURE, OTHER WAS FROZEN AND
FORM AD FEW DAYS LATER. WE STILL
SEE THAT THEY ARE FLUFFY BUT
SHOW NEURITE OUTGROWTH AND WE
ARE LOOKING LEK ROW PHYSIOLOGY
TO OPTIMIZE PROCESS --
ELECTROPHYSIOLOGY TO OPTIMIZE
THE PROCESSES. THIS IS TO REALLY
HAVE AN OPPORTUNITY TO SEND THEM
OUT, AT ROOM TEMPERATURE WE CAN
SEND THEM OUT AND CERTAIN CASES
SEVEN DAYS OF TRANSPORT DID NOT
DAMAGE THEM, THEY CAN BE USED
FOR FURTHER EXPERIMENTS. SO FOR
EXAMPLE, WE ARE DELIVERING BRAIN
ORGANOIDS FOR RABIES RESEARCH IN
BELGIUM AND AUSTRALIA FOR
COGNITIVE FUNCTION. AND WOULD BE
TO BRING THEM ALSO IN OTHER
ENVIRONMENTS OF RESEARCH. I
WOULD LIKE TO SAY FEW WORDS ON
QUALITY. I'M ONE OF THE
PROPOINTS OF CELL CULTURE
PRACTICES IN VITRO REPORTING
STANDARDS, AND VALIDATION,
HAVING THE VALIDATION BODY OF
THE EUROPEAN COMMISSION. AS
APPEARS IN 1996, I WANT TO BE
SHORT PUBLISHED IN 2005 AS A
GUIDANCE FROM EUROPEAN
COMMISSION. WE TEAMED UP IN
2015, THIS GUIDANCE DOESN'T
INCLUDE STEM CELL MODELS,
MICROPHYSIOLOGICAL SYSTEMS, SO
WE TEAMED UP WITH VARIOUS
ORGANIZATIONS AND HAD TWO
WORKSHOPS AND THESE ARE WORKSHOP
REPORTS, WHICH ARE AVAILABLE IN
THE MEANTIME AND WE THEN FINALLY
PUBLISHED IN JANUARY OF THIS
YEAR GUIDANCE DOCUMENT OF 350
COMMENTS FROM STAKEHOLDERS MORE
THAN 100 DIFFERENT STAKEHOLDERS
COMMENTING ON THIS AND THIS IS
NOW NEW GUIDANCE DOCUMENT AND WE
ARE ENTERING DISSEMINATION PHASE
TRYING TO CONVINCE FUNDING
BODIES AND JOURNALS TO ACCEPT
THIS AS A NEW STANDARD. WE ALSO
INITIATED ALREADY WORK TOWARDS
REPORTING BECAUSE WE
REPRODUCIBILITY PROBLEMS IN OUR
FIELD ARE NOT DUE TO US BUT
BETTER REPORT E BAD REPORTING
AND DIFFICULT TO REPRODUCE. WE
HAVE SOME EXAMPLES LIKE THE IN
VITRO TOXICOLOGY FEMME PLATE,
THIS IS ONGOING AND HOPE TO
EXTEND TO MICROPHYSICAL GENETIC
SYSTEMS. WITH THIS I WOULD LIKE
TO CLOSE. USING (INDISCERNIBLE)
VERY SUITING QUOTE, THE
DIFFICULTY LIES NOT IN THE NEW
IDEAS BUT ESCAPING FROM THE OLD
ONES. THANK YOU VERY MUCH.
>> THANK YOU VERY MUCH, THAT WAS
WONDERFUL TALK. VERY INTERESTING
TOPIC. I LOOK FORWARD TO GETTING
THE QUESTIONS AT THE END OF THE
PRESENTATIONS. I WOULD LIKE TO
INTRODUCE OUR LAST SPEAKER FOR
THIS SESSION. DR. ROSADO
OLIVIERI, VICE PRESIDENT OF
RUMIVIRO. DR. ROSADO RECEIVED
STEM CELL BIOLOGY FROM HARVARD
MEDICAL SCHOOL AND MIT
POST-DOCTORAL RESEARCH WITH DR.
(INAUDIBLE) AT ROCKEFELLAR
UNIVERSITY. HE DEVELOPED AN
INNOVATIVE STEM CELL BASED
PLATFORM TO TRACK INFECTION BY
RESPIRATORY VIRUSES AND IDENTIFY
THERAPEUTIC FOR COVID-19 AND IN
COLLABORATION WITH DR. CHARLIE
RICE. HE ALSO SPEARHEADED
CREATION OF RUMIVIRO WHICH IS
FIRST IN CLASS BROAD SPECTRUM
ANTIVIRAL THERAPEUTICS FOR
RESPIRATORY VIRUSES AND VIRUS
INDUCED ACUTE KIDNEY INJURY. THE
TITLE OF HIS TALK TODAY IS
STANDARDIZE HUMAN MINI ORGAN ON
MICROCHIPS TO IDENTIFY ANTIVIRAL
THERAPEUTICS AT SCALE. THE FLOOR
IS ALL YOURS.
>> THANK YOU. THANK YOU SO MUCH
FOR THE ORGANIZERS FOR INVITING
ME TO SUCH AN IMPORTANT FORUM,
TODAY IT IS MY PLEASURE TO TELL
YOU ABOUT OUR COMPANY RUMIVIRO
AND HOW WE ARE USING A
PSEUDOAPPROACH TO WHAT'S
DESCRIBED MANY TIMES WITH STEM
CELL BASED MODELS TO RECAPITATE
LATE BUT THIS TIME FULLY
STANDARDIZED APPROACHES TO
DERIVE ORGANS TO IDENTIFY
UNDEVELOPED THERAPEUTICS OF
SCALE. OF COURSE THE WAY WE DO
OUR WORK IS THE FACT THAT
BECAUSE OF THE HUGE BURDEN VIRUS
ON GLOBAL HEALTH NOT ONLY COVID
PANDEMIC BUT WITH MANY OTHER
VIRUSES INCLUDING RESPIRATORY
VIRUS AS WELL AS MANY OTHERS
THAT STILL COST A LARGE FRACTION
OF HUMAN DISEASE. THE MAIN
CHALLENGE WITH VIRUSES IN
GENERAL IS THAT FOR MANY OF
THEM, THERE IS COMPLETE ABSENCE
OF TREATMENT OR VACCINES AND
EVEN WHEN THERE ARE VACCINES FOR
MANY -- FOR SOME THERE IS ALWAYS
THESE ISSUES OF EMERGENCE OF NEW
VARIANTS WHOSE RES ISTANT
MUTATIONS IN SYSTEMS AGAIN
CURRENT -- THIS HIGHLIGHTS THE
NEED FOR NOVEL ANTIVIRAL DRUG
DISCOVERY PARADIGM THAT CAN LEAD
TO NEW THERAPEUTIC STUDIES TO
TACKLE VIRUSES THAT ARE MAY
CAUSE PANDEMIC, THIS WHERE I
THINK MODELS MAY PLAY A BIG ROLE
IN THE NEAR FUTURE. I THINK THE
CHALLENGE WITH ANTIVIRAL DRUG
DEVELOPMENT IS THE EFFORTS THAT
HAVE BEEN DEPLOYED ARE ACTUALLY
TARGET BASE WHERE DRUG IS
DEVELOPED AGAINST SPECIFIC VIRAL
TARGET OR HOST FACTOR, AND THE
CHALLENGE WITH THIS IS THAT
TARGETS ARE JUST A FEW AND HOST
FACTORS ARE TYPICALLY ELUSIVE
FOR MANY VIRUSES THAT ARE OF
CONCERN SO THERE IS A QUESTION
AS TO HOW SPECIFIC TARGET FOR
THERAPEUTIC DEVELOPMENT AND HOW
DO YOU PRIORITIZE THEM. ON THE
OTHER END OF THE SPECTRUM YOU
CAN THINK ABOUT SUB BASE
APPROACHES FOR NON-KINASE CELLS
HAS A WAY OF IDENTIFYING
ANTIVIRAL ACTIVITY AGAINST
VIRUSES, AND I THINK THE
CHALLENGE HERE IS THE FIELD HAS
ADOPTED THE CANCER CELL LINES AS
STANDARD FOR TESTING
THERAPEUTICS IN PRE-CLINICAL
ASSAYS, IN DRUG DISCOVERY AND
DRUG TESTING PARADIGMS. THE
ISSUE WITH THIS IS THAT THESE
CELL LINES LACK ORGAN FACILITY
AND WE HAVE HIGHLIGHTED MANY
TIMES DURING THE LAST FEW DAYS
THEY ALSO DISPLAY KEY
DIFFERENCES IN BIOPATHWAYS.
IMPORTANT CHALLENGE REALLY
IMPORTANT SPECIFICALLY IN
CONCEPT OF COVID-19 WHERE MANY
DRUGS WE ARE IDENTIFIED IN DRUG
REPURPOSING EFFORTS YOU HAVE
SEEP THESE CELL LINES ENDED UP
FAILING CLINICAL TRIALS DUE TO
NON-PHYSICAL MECHANISM OF ACTION
AS IT WAS HIGHLIGHTED IN THIS
RECENT STUDY BY THE GROUP OUT OF
UCSF. THIS AGAIN HIGHLIGHTS THE
NEED FOR NEW NOVEL TISSUE BASE
MODELS FOR NOT ONLY ANTIVIRAL
TESTING BUT ALSO ANTIVIRAL DRUG
DISCOVERY. TODAY I WANT TO TELL
YOU ABOUT HOW WE PROPOSE TO
REACH THAT (AUDIO GAP) AND
APPROACH THE GENERATE TARGET
TISSUE FROM STEM CELLS
SUSCEPTIBLE TO INFECTION HOW
THIS MANY ORGANS DERIVED FROM
PATIENT LIKE RESPONSES TO VIRAL
INFECTION. AND HOW WE CAN ALSO
PERFORM HIGHLY QUANTITATIVE AND
STANDARDIZE HIGH LIEU PUT SCALE
AND I THINK WHAT IS REALLY
UNIQUE ABOUT THIS APPROACHES ARE
NOW FOR THE FIRST TIME WE CAN
REALLY PLAY HOST VIRAL
INTERACTIONS AT CENTER OF DRUG
DISCOVERY DRUG DEVELOPMENT
PIPELINE. DRUG DISCOVERY WE ARE
NOW UTILIZING A NEW ERA OF
CLINICAL MODELS BY COMBINING A
HIGH THROUGH PUT UNBIASED
APPROACH THAT IS THE BEST MANNER
TO DISCOVER NEW MECHANISM OF
ACTION AND ALSO NEW THERAPEUTIC
HYPOTHESIS, AND COMBINE THAT
WITH DERIVATION OF SYNTHETIC
HUMAN TISSUE WHICH YOU HAVE
HIGHLIGHTED MANY TIMES LAST TWO
DAYS AS BEING THE BEST CLINICAL
MODS OF HUMAN PHYSIOLOGY BUT
ALSO THE BEST PREDICTORS OF
HUMAN EFFICACIES.
(INDISCERNIBLE) ALLOWS US TO
BUILD THE RIGHT TISSUE, WE DO
THAT WITH MICROPATENT TECHNOLOGY
THAT ALLOWS US TO STANDARDIZE
TRANSITION WITH STEM CELLS, WE
DO SO BY CREATING THOUSANDS OF
STEM CELLS IN THIS BY EMBRYONIC
STEM CELLS IPSCs THAT WE CAN
GROW ON VERY WELL DESIGNED
SPECIFIC SIZE MORPHOLOGIES, THIS
IS WHAT IS KEY TO OUR APPROACH
BECAUSE IT IS MANY SOMETHING YOU
CANNOT DO TO THE EXTENT STEM
CELL CULTURES THAT ARE VARIABLE
AND HIGHLY (INAUDIBLE) IN NAT
NATURE. ANOTHER (INAUDIBLE)
WORKING WITH THIS UTILIZATION
APPROACH WE HAVE ALSO SHOWN THAT
WHEN STEM CELL IS DIFFERENTIATE
UNDER THIS COLONIES THEY
DIFFERENTIATE ON ORGANOIDS TO
CREATE COMPLEX TISSUES AND YOU
WILL SEE IN MY TALK LATER TODAY
THAT THESE TISSUES ARE --
SELF-ORGANIZE INTO CLOSE TO MY
MIMICKING HUMAN COUNTER PARTS.
THIS I LIGHT HOW THESE
APPROACHES NOT ONLY ALLOWS US TO
STANDARDIZE THE SUBSTANTIAL
DIFFERENTIATION BUT ALLOWS US TO
GENERATE SELF-ORGANIZED ORGANS
THAT RESEMBLE HUMAN COUNTER
PART. THIS IS AN IMPORTANT STEP
TOWARD DEPLOYMENT OF THESE
MODELS FOR DRUG DISDVRRY BECAUSE
AS WE HAVE DISCUSS MANY TIME
THROUGH THE ORGANOID HAVE BEEN
REALLY IMPORTANT FOR SO MANY
DIFFERENT ASPECTS OF VIRAL
DISEASE BUT RIALITY IS THEY ARE
NOT -- REALITY IS THEY ARE NOT
WELL ENOUGH FOR DRUG DISCOVERY
LARGE SCALE AS WE HAVE ASSESSED
MANY TIMES AND THE REASON FOR
THAT IS THAT THESE VARIABLE,
THEY ALLOW US TO PERFORM TARGET
OF ANY GIVEN TISSUE BECAUSE THIS
IS MICROPATENT TECHNOLOGY THIS
ALLOWS THOUSANDS OF MINI ORGANS
IN PARALLEL IN DOING SO WE CAN
CREATE LARGE APPROACHES TO
DEFINE SET OF QUANTITATIVE
FENTYPICS A ASSOCIATED WITH ANY
GIVEN DISEASE, AND (INAUDIBLE)
ALLOWS US TO STREAM LARNING
CLINICAL RESCUE IN GIVEN DISEASE
ASSOCIATED PHENOTYPIC SIGNATURE
ANOTHER ASPECT OF OUR WORK HAS
BEEN REALLY IMPORTANT IS THAT WE
TRULY TAKE VERSATILITY OF STEM
CELLS BECAUSE NOW WE CAN THINK
APPLYING THIS TO ANY ORGAN AND
TISSUE IN OUR BODY AND TISSUES
THAT ARE HIGHLY SUSCEPTIBLE TO
INFECTION. TODAY I'M -- ALSO
NEXT FEW MINUTES ON THE
(INAUDIBLE) SYSTEM WE DEPLOYED
TO STUDY VIRAL INFECTION IN THE
LUNG. WE ALSO HAVE ACCESSING
DEVELOPMENTS TO DERIVE TISSUES
OF THE LIVER KIDNEY AS WELL AS
BRAIN, WILL ALLOW US TO NOT ONLY
MODEL INFECTION BY THIS PERSON,
SPECIFIC ORGAN TISSUE CONTEXT
BUT ALSO POTENTIALLY IDENTIFY
THERAPEUTIC AT SCALE FOR ANY OF
THESE VIRUSES. HIGHLIGHTS HOW
THIS PLATFORM ASSESS TISSUE THAT
IS HIGHLY REPRODUCIBLE AND
AMENABLE TO HIGH THROUGH PUT IN
MANY DIFFERENT DISEASE CONTEXT.
TODAY THIS WORK WAS REALLY
STARTED BECAUSE OF THE PANDEMIC
WHEN I WAS POST-DOC AT
ROCKEFELLAR UNIVERSITY IN
COLLABORATION -- WHEN WE STARTED
THE PANDEMIC -- WHEN THE
PANDEMIC STARTED WE WANTED TO
DEVELOP MODEL SYSTEM THAT WILL
ALLOW US TO GENERATE MINI LUNG
TISSUE IN STANDARDIZED WAY, AND
WE CAME UP WITH HISTORICAL DATA
THAT I ALLOWS US TO GENERATE
MICROPARTICLES THAT DISTRIBUTE
TISSUE STRUCTURES, THAT CONTAIN
TWO TISSUE TYPES INCLUDING
AIRWAY CELLS ALVEOLAR CELLS FORM
IN THIS CONTINUOUS EPITHELIA
STRUCTURE FORMING ON TOP OF THE
-- WAS FASCINATING ABOUT THIS
APPROACH IS THIS PROCESS TAKES
13 DAYS. ANOTHER ASPECT OF THIS
WORK IS THAT WE HAVE SHOWN THIS
TISSUES ARE CLOSELY MANIPULATING
HOW TISSUES LOOK LIKE IN
PATIENTS ORGANS AND JUST TO
ILLUSTRATE DIAGRAM HOW ANY HUMAN
LOOKS LIKE AND I HOPE YOU CAN
APPRECIATE HUMAN PATIENTS ALSO
HAVE SAME STRUCTURE WITH SAME
PROXIMAL DISTAL ORGANIZATION OF
AIRWAY ALVEOLAR CELLS. THIS
HIGHLIGHT IT IS UTILITY OF OUR
PLATFORM TO NOT ONLY GET ACCESS
TO PHYSIOLOGICAL LUNG TISSUE BUT
ALSO TISSUES THAT ARE MIMICKING
HUMAN COUNTER PARTS IN BOTH
TISSUE COMPLEXITY AND TISSUE
MORPHOLOGY. WE -- IT STARTED TO
DEVELOP ASSAYS ABOUT US TO
REALLY -- BY SARS COV-2 THIS IS
INITIALLY WE EFFICIENT DERIVED
ISOLATE OF SARS COV-2. WE SAW
THIS SYSTEM WAS HIGHLY
SUSCEPTIBLE TO SARS COV-2
INFECTION AS YOU CAN SEE HERE IN
THIS MICROGRAPH TO
REPRESENTATIVE -- WE DON'T SEE
INFECTION IN BOTH ALVEOLAR
AIRWAY CELLS. THIS PLATFORM IS
TRULY UNABLE TO HIGH RESOLUTION
CONFOCAL IMAGING YOU CAN SEE
HERE IN THIS VIDEO WHERE WE CAN
TRACK CELL TISSUE WITHIN A
MINUTENY, BECAUSE IN THE
MICROBIOME WE CAN TRACK IN MANY
THOUSANDS OF PARALLEL AND
IDENTIFY THE DISEASE FEATURE
WITH INFECTION BY VIRUS IN THIS
CASE SARS COV-2. WE WERE REALLY
EXCITED ABOUT THIS HE CAN
TECHNOLOGY, WE WANTED ASSAYS
THAT WOULD ALLOW US TO PERFORM
HIGHLY QUANTITATIVE AANALYSIS OF
SPECIFIC PROCESSES OF THE VIRAL
LIFE CYCLE, WE WERE LOOKING AT
VIRUS IDENTIFY POTENTIALLY TUSH
SHOE SPECIFIC DIFFERENCES IN
ACTIVITY TO INFECTION BETWEEN
AIRWAY ALVEOLAR CELLS WE SAW
THAT ALVEOLAR CELLS ARE MORE
SUSCEPTIBLE TO INFECTION THAN
AIRWAY CELLS, WE ALSO DEVELOPED
ASSAYS THAT ALLOWS US TO LOOK AT
REPLICATION TRANSMISSION OF
VIRUS, THIS TRANSMISSION
PARAMETER WE SEE INCREASE IN
NUMBER OF INFECTED CELLS OVER
TIME. WE SEE REFLECTS AFTER
REPLICATION OF VIRUS IN LUNG
TISSUE. WE CAN QUANTIFY HIGH
LEVELS OF APOPTOSIS IN ACUTE
PHASE OF BOTH INFECTION AND
TRANSMISSION. THIS IS REALLY THE
MOST -- SORRY ABOUT THAT. THIS
IS NOT TO ONLY RECAPITULATE
ENTIRE LIFE CYCLE OF THE VIRUS
BUT ALSO APPLY HIGHLY
QUANTITATIVE ANALYSIS THAT
ALLOWS US TO REALLY QUANTIFY
EVERY STEP INDIVIDUALLY. IN
MANY LUNGS CYTOKINES. OF COURSE
OUR WE ARE EXCITED BECAUSE THIS
IS IS -- SUN STATISTIC
PHYSIOLOGICAL RESPONSES WE ARE
INTERESTED IN UNDERSTANDING
WHETHER THIS SYSTEM WAS
RECAPITULATED IN ASSOCIATED WITH
COVID (INAUDIBLE) AND DO THEY
PERFORM GENE EXPRESSION ANALYSIS
AND COMPARISONS WITH POSTMORTEM
TISSUE OF COVID PATIENTS THIS
COHORT OF ALMOST 40 PATIENTS,
THAT DIED BECAUSE OF COVID. H
THIS IS LUNG TISSUE RECOVERED
FROM THEM. WE ARE LOOKING AT
CORRELATION, BETWEEN OUR MODEL
AND PATIENT SAMPLES AND WHAT WE
SAW IS THAT AMONG TOP EXPRESSED
GENES YOU CAN SEE HERE IN THE
HEAT MAP, THERE IS HIGH
CORRELATION BETWEEN GENES IN
LUNG BIOPSY INFECTED LUNG BIOPSY
COMPARED TO INFECTED LUNG WHERE
YOU SEE GENES HAVE SIMILAR
EXPRESSION PATTERNS. ALSO
INTERESTING IN CELL LINES
RECAPITULATED THESE GENES DO NOT
CHANGE IN EXPRESSION OF -- TO
INFECTION IN SIMILAR WAY. TO
TALK -- TAKE A STEP FURTHER WE
PERFORM ANALYSIS TO LOOK AT
ENRICHMENT OF THIS PATIENT
ASSOCIATED GENE EXPRESSION
POLYMERS. WE FOUND THAT OUR
MINIMUM (INAUDIBLE) DISPLAY HIGH
ENRICHMENT FOR THESE ENGINE
EXPRESSION PROGRAMS CORRELATED
WITH COVID-19 IN PATIENTS. THIS
SOMETHING NOT RECAPITULATED
USING STANDARD CELL LINES,
APOLOGIZE FOR TYPO, WE DIDN'T
ENRICHMENT OF GENE EXPRESSION
PROGRAMS, THIS IS DEMONSTRATING
THAT OUR PLATFORM ANALYSIS TO
CLOSELY MIMIC PATIENT LIKE COVID
RESPONSES, IN A WAY UNPARALLEL
TO ANY OTHER CELL BASE RESEARCH
TOOL THAT ARE PLOYED AS STANDARD
AND THERE IS PRE-CLINICAL
TESTING OF PRE-CLINICAL
VALIDATION OF COMPOUNDS. WE
HAVE ACCESS TO A SYSTEM THAT WE
CAN USE TO INTERROGATE HOST
TARGETS CRITICAL FOR (INAUDIBLE)
WE ARE EXCITED ABOUT MINING DATA
FROM CRISPER SCREENS THAT
IDENTIFY GENES ESSENTIAL FOR
INFECTION BY SARS COV-2 OR ALSO
ENDEMIC CORONA VIRUS HERE
COLLABORATING WITH CHARLIE RICE
WHERE WE ROOK AT GENES THAT
IDENTIFY ESSENTIAL GENES FOR
SARS COV-2 AND WE TRIED TO
VALIDATE WHICH OF THEM HAD WORK
RELATED INFECTION IN THE
NEURONAL SYSTE SYSTEM. WE FOUNDE
GENE THAT WAS HIGHLY CORRELATED
WITH SARS COV-2 NOT ONLY IN THE
LUNG SYSTEM BUT ALSO IN PATIENT
SAMPLES WE SAW THAT WAS HIGHLY
UPREGULATED FROM SARS COV-2
INFECTION. AND NOW THIS GENE WAS
INTERESTING AS A DRUGGABLE
TARGET, THERE ARE -- WE ARE
EXCITED ABOUT USING OUR MODEL TO
TEST WHETHER THERAPEUTICS AND
ACTIVITY WOULD HAVE ANY EFFECT
ON SARS COV-2 INFECTION IN THE
LUNGS AND REALLY THAT WAS
FASCINATING TO FIND INHIBITOR OF
THIS GENE DEMONSTRATED POTENT
INDIVIDUAL INHIBITORY ACTIVITY
AGAINST SARS COV-2 IN OUR ASSAY
AGAIN THIS CLEARLY HIGHLIGHTS
GENETICS BUT ALSO DRUGS GIVEN ON
TESTING APPROACHES CAN BE USED
TO DISCOVER NEW HOST BASE
MECHANISM OF ACTION THAT
ACTUALLY ARE FOCUSING ON TARGET
THAT HOST RESPONDS PRODUCING
VIRAL INFECTION. E O WE WANTED
TO TRANSFER THIS TO HIGH PUT
SETTING SO WE TRANSFER TO 96 AND
384 WELL PLACE WHERE EVERY WELL
OF THE MULTI-WELL PLATE CONTAIN
TENS OF QUANTITIES OF LUNG
TISSUE HIGHLY SUSCEPTIBLE TO
VIRUS INFECTION. SARS COV-2 AS
PROOF OF CONCEPT FOR TESTING
THIS PLATFORM, WE COLLABORATED
WITH (INAUDIBLE) WILSON LAB AT
ROCKEFELLAR UNIVERSITY WHERE WE
WANTED TO TEST MONOCLONAL
ANTIBODIES ISOLATED FROM
CONVALESCENT PLASMA
PUBLICATIONS. HERE IS ONE OF
ANTIBODIES 121 WHICH IS
CURRENTLY IN CLINICAL TRIALS AT
THIS POINT, THAT SHOW PROMISING
POTENT ANTIVIRAL NEUTRALIZING
ACTIVITY AGAINST SARS COV-2 IN
OUR MINIMUM ASSAY. WE CAN
BECAUSE THIS IS HIGH THROUGH PUT
ASSAY WE CAN NOW PERFORM HIGHLY
QUANTITATIVE ANALYSIS IN THIS
CASE ANTIBODY POTENCY AND
EFFICACY, YOU CAN THINK OF USING
THIS HIGH THROUGH PUT PLATFORM
TO ALSO TEST POTENTIALLY OTHER
CANDIDATE ANTIVIRAL
THERAPEUTICS. JUST DEMONSTRATE
THIS PLATFORM, I WANT TO SPEND
TIME SHOWING YOU THE PLATFORM
WHERE WE SEE THAT DEMONSTRATES
HIGH SENSITIVITY TO LEVEL OF
SARS COV-2 INFECTION SO WE SEE
IN THIS SOLUTION ASSAY, THE
ASSAY THE DATA WAS VERY ROBUST
AND IN THIS CASE CORRESPONDS TO
SINGLE ORGAN SO QUANTIFYING MANY
AT A TIME. WHEN WE LOOK AT
SEQUENCE SCORES BETWEEN POSITIVE
AND NEGATIVE CONTROLS WE SEE
WEIGHTED .5. WE ALSO SERVE
COEFFICIENT VARIANT LESS THAN
20% WITH ASSAY METRICS THAT
SPEAKS A LOT AS TO THE ASSAY
STANDARDIZATION OF THIS ASSAY.
ON HIGH THROUGH PUT SCALE. WE
HAVE ALSO DEPLOYED PLATFORM TO
START TESTING AND IDENTIFYING
CANDIDATE THERAPEUTICS
PARTICULAR VIRUSES DEMON
INVESTIGATING SHOWING YOU OUR
RECENT RESULTS WITH OUR WORK
WITH RSV SHOWING YOU HEAT MAP OF
COMPOUND TREATED WELL PLATE WITH
(INAUDIBLE) ON THE OUTSIDE AND
YOU CAN CLEARLY IDENTIFY VERY
POTENT ACTIVITY AGAINST RSV, NOW
YOU CAN REPEAT THE PROCESS WITH
THOUSANDS AND THOUSANDS OF
COMPOUNDS, EVEN IDENTIFY THOSE
THAT HAVE VERY POTENT INHIBITION
ACTIVITY AGAINST RSV IN THIS
CASE WE WERE EXCITED ABOUT THIS
19 COMPOUNDS DEMONSTRATED
INHIBITOR ACTIVITY AGAINST RSV
GREATER THAN 80%. THIS CREATES
FOR US TO STREAM LARGE CLINICAL
LIBRARIES TO IDENTIFY ANTIVIRAL
THERAPEUTICS FOR CANDIDATE
VIRUSES AS WELL. EXCITING PART
OF OUR WORK IS HAS THE ABILITY
TO (INAUDIBLE) VIRUSES BECAUSE
ALSO CONTAINS OLDER VIRUSES AND
WE HAVE SHOWN THIS ARE
SUSCEPTIBLE TO INFECTION BY
OTHER ENDEMIC VIRUSES HERE AS
WELL AS RSV, SOME INFLUENZA AND
THIS AGAIN IS DIAGNOSTIC AND
RECAPITULATE INFECTION BY ANY
GIVEN VIRUS, THINK ABOUT
DEVELOPING HIGH THROUGH PUT DRUG
DISCOVERY CAMPAIGN GIVEN THE
HIGH THROUGH PUT ASPECT OF OUR
WORK. WE ARE TAKING THIS
APPROACH FURTHER. WE ARE EXCITED
(INAUDIBLE) IN HOUSE DISCOVERY
PROGRAMS WE ARE ACTIVELY USING
THE SYSTEM TO IDENTIFY BROAD
SPECTRUM ANTIVIRALS FOR
RESPIRATORY VIRAL INFECTION AND
THAT IS FOR ALL MAJOR CLASSES OF
VIRUS PREFERENTIALLY IN THE LUNG
AND WE ARE TAKING UP PLATFORM TO
IDENTIFY BROAD SPECTRUM
ANTIVIRUS FOR ACUTE INJURY, THIS
IS HIGHLIGHTING THE ACTIVITY OF
3D TISSUE MODELS AND FULLY
STANDARDIZED THAT CAN ENABLE
IDENTIFICATION OF NEW THERAPY
BASED ON TISSUE TARGETED HOST
BASE THERAPEUTICS. WITH THAT I
WOULD LIKE TO SUMMARIZE WHAT I
PRESENTED SO FAR, AS I SAID THE
MAIN MUTATIONS ARE CLEARLY OF
ANTIVIRAL DEVELOPMENT IS LACK OF
STANDARDIZE PHYSIOLOGICAL
MODELS. I TOLD YOU HOW MANY
ORGANS IN GENERAL OFFER LIMITED
ACCESS TO STANDARDIZE PATIENT
LIKE TISSUE WE CAN USE TO MODEL
INFECTION LIKE VIRUSES AND AT
THE SAME TIME PERFORM HIGHLY
QUANTITATIVE ANALYSIS OF NOT
ONLY SPECIFIC PROCESSES OF VIRAL
PROCESSES OF THE VIRAL LIFE
CYCLE BUT ALSO TEST THERAPEUTICS
AND HOW THEY CAN MEDIATE THESE
PROCESSES. I ALSO TOLD YOU ABOUT
HOW THIS HIGH THROUGH PUT
PLATFORM IS HIGHLY SENSITIVE AND
CAN BE USED TO IDENTIFY
CANDIDATE ANTIVIRAL AGAINST
PARTICULAR VIRUS. THIS CLEARLY
HIGHLIGHTS HOW THIS MODEL WILL
ENABLE IDENTIFICATION OF FIRST
IN CLASS MOLECULES FOR VARIOUS
VIRUSES, BASED ON THIS
STANDARDIZED 3D TISSUE MODEL.
WITH THAT I WOULD LIKE TO END MY
TALK AND LOOKING FORWARD TO OUR
Q&A SESSION. THANK YOU.
>> THANK YOU VERY MUCH.
FANTASTIC TALK. I WOULD LIKE TO
ASK SPEAKERS TO TURN ON CAMERAS
AND THEIR MICROPHONES. WE HAVE
15 MINUTES FOR Q&A. I WILL START
WITH THE QUESTION FROM AUDIENCE
FOR THOMAS. WHAT DO YOU THINK
ABOUT THE FACT THAT THERE IS NO
INTERFERON RESPONSE IN THE BRAIN
ORGANOIDS AFTER SARS COV-2
INFECTION? THOMAS, THAT IS A
QUESTION FOR YOU THE BRAIN
ORGANOID SYSTEMS. QUESTION FROM
THE AUDIENCE ABOUT THE LACK OF
INTERFERON RESPONSE. AFTER SARS
COV-2 INFECTION. THAT IMPACT
RATTLE YOU ARE SEEING OR IS THAT
PART OF DOES THAT IMPACT THE
NATURAL PHYSIOLOGY OF INFECTION?
>> I CANNOT SPEAK TO THIS FROM
-- NOT ADDRESS THIS ISSUE. THERE
IS OBVIOUSLY NO INTERFERON
PRODUCING CELL MEASURE, BUT WE
HAVE ANOTHER CONTEXT ALSO
LYMPHOCYTIC CELLS AND SEPSIS
MODELS AND POSSIBLE ALSO TO
ENGAGE OTHER CELL TYPES WHICH
MIGHT INVADE AND CONTRIBUTE
THIS. I MIGHT BE COMPLETELY OFF
THIS IS NOT MY AREA OF RESEARCH
>> NEXT QUESTION FROM THE
AUDIENCE, COMPARED TO AIR LIQUID
INTERFACE MODEL HAVE YOU ALSO
VALIDATED THE OTHER CELL
COMPONENTS? YOUR MODEL?
SILLIATION AND MUCOUS
PRODUCTION?
>> WE DID AND I DIDN'T HAVE TIME
TO DISCUSS IN DETAIL, RNA
SEQUENCING AND HOW IDENTIFY THE
CELL TYPES IN THESE LUNGS, WE
FOUND CELL TYPES IN AIRWAY
COMPARTMENT MOSTLY CELLS MOSTLY
CELLS THAT HAVE CILIA OPPOSED TO
THE APICAL SITE AND THERE IS
THREE MAJOR CELL TYPES IN STEM
CELL LIKE THAT WAS IDENTIFY IN
TISSUE AS WELL AS WELL AS EARLY
TYPE 1 AND 2 PNEUMOCYTE SO THOSE
ARE THE BEST WORK WE HAVE DONE
AND THOSE MUCOUS CELLS MUCOUS
PLUCKS IS SOMETHING WE HAVEN'T
DONE AND WE DON'T HAVE A LOT OF
CELLS AND A LOT OF MUCOUS CELLS
IN OUR MODEL SYSTEM WHICH IS OF
COURSE LIMITATION BUT WE DO HAVE
USUAL SUSPECTS THE MAJOR
(INAUDIBLE) IN THE LUNG SYSTEM.
ONE AREA I WAS INTERESTED IN IS
WHAT ARE SOME OF THE WHAT ARE
TECHNICAL REGULATORY HURDLES FOR
ADOPTING THESE 3D CULTURE MODEL
SYSTEMS AND DRUG DEVELOPMENT?
EVI AND I IN NICOLE MIGHT BE
ABLE TO ADDRESS THIS BASED ON
YOUR EXPERIENCES.
>> I'LL DEFER TO EVI. BEING THE
I DO A LOT OF WORK IN SUPPORT OF
REGULATORY AGENCIES. PLEASE GO
I MED THEN I WILL ADD MY
THOUGHTS.
>> SO IN TERMS OF HURDLES, I CAN
ONLY TALK ABOUT WHAT WE SEE IN
OUR BRANCH AND OUR DIVISION.
FIRST I HAVEN'T SEEN MANY OF
THESE INNOVATIONS TO SUPPORT OUR
PRODUCT. HAVE NOT MADE IT DOWN
THROUGH THE REGULATORY PATH. I
DON'T KNOW WHAT THE HURDLE IS.
MAYBE THE THINKING IS WE DON'T
HAVE ACCEPTANCE OR THE
CAPABILITY OF REVIEWING THIS
DATA, I AM NOT SURE. BUT AGAIN I
WILL SAY THAT WE HAVE SEEN MUCH
OF THIS. ALSO ORB POINT IS -- WE
TEND TO NEUTRALIZATION ASSAYS
HAVE BEEN TRADITIONALLY USED AND
WE HAVE ACCEPTED THEM AS GOOD
INDICATION OF ACTIVITY BUT FOR
-- AS MORE AND MORE
SOPHISTICATED AND MORE
COMPLICATED PRODUCTS ARE COMING
AND BEING DEVELOPED WE MAYBE
NEED TO KNOW MORE AND SEE MORE.
THE OTHER THING IS, AS WAS OFTEN
DISCUSSED IN CONTEXT OF SARS
COV-2, IS THE FACT THAT FOR SOME
EMERGING VIRAL DISEASE THE
ANIMAL MODELS ARE NOT THERE. AND
BECAUSE IN THIS CASE
MICROPHYSIOLOGICAL SET UP WOULD
BE A REPLACEMENT, WOULD BE A
TEST THAT IS MORE APPLICABLE TO
THE SITUATION SO IN THAT
SCENARIO WE WOULD ENCOURAGE SUCH
STUDIES TO COME IN THE DOOR. I
AGREE. BUT ALSO THERE IS A
DISCONNECT AT MOMENT. LAST WEEK
I WAS AT THIS INCREDIBLE MEETING
THE TEAM FROM JOHNS HOPKINS
HELPED CO-ORGANIZE OR ORGANIZED
AND HELPED BY OTHER INSTITUTIONS
AS WELL, MICROPHYSIOLOGICAL
SYSTEM WORLD SUMMIT AND THERE
WAS INCREDIBLE SCIENCE BEING
PRESENCED. THERE IS A LITTLE BIT
OF DISCONNECT BETWEEN REALLY
EXCITING CUTTING EDGE SCIENCE
AND THE SORT OF REAL GROUNDED IN
REALITY PRACTICALITY NEEDS OF
REGULATORY DECISION MAKERS. SORT
OF ONE OF THE PIECES OF FEEDBACK
WE GOT WHEN WE THROUGH THE CORE
WORKING GROUP LEADERSHIP OF THAT
GROUP MET WITH EUROPEAN
REGULATORS PARTICULARLY WITH
EUROPEAN MEDICINE AUTHORITY AND
THE SAFETY WORKING PARTY WAS
SORT OF THE NEED TO HAVE MODELS
LIKE THE FOCUS INITIALLY FOR
ESTABLISHING REGULATORY UTILITY,
AND CONFIDENCE, SHOULD BE ON
SIMPLE SINGLE ORGAN MODEL
APPLIED TO SAFETY PHARMACOLOGY
END POINTS RATHER THAN THESE
MUCH MORE COMPLICATED BUT VERY
SEXY BODIES ON A CHIP, MULTIPLE
DIFFERENT TISSUES, DAISY CHAINED
TOGETHER TO REPEAT SIX MONTH
DOSE STUDIES ALL REALLY EXCITING
AND NEEDED TO MOVE THE FIELD
FORWARD BUT YOU CAN'T START WITH
THAT. IN MANY TERMS OF
REGULATORY CONSIDERATIONSS.
ACCEPTANCE.
I WILL SAY THERE WAS A EXAMPLE
THAT WAS PRESENTED LAST WEEK
WHERE THE FIRST IND APPLICATION
SUPPORTED EXCLUSIVELY BY TISSUE
CHIP DATA WAS SUBMITTED
SUCCESSFULLY TO THE FDA AND
PERHAPS THOMAS CAN EXPAND ON
THAT A LITTLE BIT.
>> IMPORTANT -- THERE THE FIRST
EXAMPLES HAS BEEN ABLE TO
SUPPORT (INAUDIBLE) THERE'S
OTHER CASES WHERE DERISKING OF
SUBSTANCES TOOK PLACE IN FORM OF
INVESTIGATIVE TOXICOLOGY AND
THINGS COULD BE CONTINUED. BUT
AT THE MOMENT WE HAVE A
SITUATION WHICH IS -- THERE IS
NOT REALLY -- NOBODY -- VERY FEW
CAN OVERSEE WHAT IT IS PRODUCING
BECAUSE SO MUCH (INAUDIBLE) THEY
NEED TO IDENTIFY SRI STANDARDS
WHAT IS THE TO GO MODELS
ROBUSTLY AVAILABLE ALSO DEPEND
ON WHAT IS COMMERCIALLY
AVAILABLE BECAUSE THIS IS WAY OF
STANDARDIZATION. WE DON'T HAVE A
SITUATION WHERE EACH AND EVERY
MODEL CAN BE VALIDATED BY EACH
AND EVERY COMPANY WHO WANTS TO
USE IT SO WE NEED THE MASTER
FILES, THE BEST PRACTICES CREATE
SOMETHING BOTH USERS AND
REGULATORS CAN RELATE TO IN THE
END. IT IS THE VALIDATION. THIS
IS WHAT WILL BE REQUIRED. THESE
ASSAYS HAVE SO COMPLEX YOU DON'T
WANT TO PRODUCE A FILE FOR THE
FDA EVERY TIME YOU WANT TO
SUPPORT AN APPLICATION.
>> THIS IS A GREAT DISCUSSION
FROM MY EXPERIENCE TOO, THERE IS
FLEXIBILITY FROM THE REGULATORY
AGENCIESND EVALUATING ASSAYS AS
LONG AS WELL VALIDATED WITH
STANDARDS AND CONTROLS AND VERY
SPECIFICATIONS YOU POINT OUT FOR
REGULATORY PURPOSES. THERE IS
HESITANCE FROM THE INDUSTRY TO
TAKE ON THAT RESPONSIBILITY.
THERE ARE OTHER EASIER WAYS TO
GET THAT DATA OR MORE STANDARD
WAYS TO GET THAT DATA, THAT IS
PREFERRED PATH BUT AS YOU SAY
THOMAS, IF THERE ARE SOME ASSAYS
THAT HAVE BEEN MORE WELL
CHARACTERIZED, VALIDATED FOR
SPECIFIC USE PURPOSES, PERHAPS
HAVING IT EITHER -- GO AHEAD.
>> THE INDUSTRY HAS TO COME
TOGETHER AND FORM CONSORTIA TO
JOIN BECAUSE IT IS MORE
EXPENSIVE AND DIFFICULT TO
EVALUATE THESE SYSTEMS AND ALSO
FEEDBACK TO THE WHAT IS NEEDED
AND DIFFERENT WEAKNESSES. THERE
IS TREMENDOUS -- I HAVE NEVER
SEEN SOMETHING LIKE WHAT THE FDA
IS DOING WITH MPS. THE DOOR WITH
THE ICELAND PROGRAM FOR
QUALIFICATION. WHICH HAS TO BE
TRIED. WE IMMEDIATE THE FIRST
MPS TO GO THROUGH PROCESS TO SEE
IF THIS FEASIBLE, WAY OF
CREATING THE TRUST WHICH IS
REQUIRED. THE ONLY THING WORD
DRUG DEVELOPMENT IS FDA, VERY
SIMPLE, 4.3% OF THE WORLD
POPULATION BUT WE CONSUME 48% OF
ALL DRUGS MARKETED IN THE WORLD
AND 64% WHAT IS SOLD ON THE
PATENT SO PHARMA COMPANIES LOOK
FOR FDA DOING. THIS IS WHY
ATTITUDE IS SUCH BREAK THROUGH
FOR ADOPTING EARLY.
>> VERY GOOD POINT. THANK YOU,
NICOLE. YOU BRING BACK VERY GOOD
POINT. THE FACT THAT THE
INDUSTRY NEEDS TO FORM THIS
CONSORTIA AND HAVE THIS DRUG, WE
HAVE EXPERIENCE AND USE THEM
BEFORE IN SARS COV-2 WAS
EXCELLENT EXAMPLE. AND MASTER
FILES GET WISE USE AND HELPED
ASSAY AS REVIEWERS OF THESE
APPLICATIONS AND HELP COMPANIES
EXAMINING THINGS FORWARD. I
THINK THAT IS A SPACE THAT COULD
BE FURTHER EXPLORED AND WORKED
ON.
>> IT IS ALSO IMPORTANT TO
CONSIDER THE FACT THAT THERE IS
OBVIOUSLY CONTEXT OF USE IS
ESSENTIALLY THE MOST IMPORTANT
THING WHEN VALIDATING AN
APPROACH AND WHEN SCIENTIFIC
CONFIDENCE AND THERE IS A
DIFFERENT BAR BETWEEN
DEMONSTRATING EFFICACY AND
DEMONSTRATING SAFETY. AT THE
MOMENT THERE IS TREMENDOUS
OPPORTUNITY IN THE EFFICACY
SPACE, SPARLY AS EVIDENCED BY
EVI'S TALK AND THE TALKS I SAW
LAST WEEK PARTICULARLY WORK
BEING DONE BY ROCHE AND OTHER
COMPANIES, IN MANY THE BIOLOGICS
SPACE. SO MANY NEW MOLECULES
COMING OUT ARE BIOLOGICS AND
THEY SIMPLY DO NOT HAVE
NON-HUMAN TARGETS. THEY JUST
DON'T HAVE THE OPTION OF USING
AN ANIMAL MODEL TO TEST THEM
BECAUSE THE ANIMAL SPECIES
DOESN'T POSSESS THE TARGET
ENGINEERED FOR. SO I THINK THAT
IS AN AREA WHERE THERE IS REALLY
EXCITING OPPORTUNITY AND
ABSOLUTE REQUIREMENT TO USE
HUMAN BASED SYSTEMS TO ADDRESS
THOSE PARTICULAR BIOLOGICS.
>> I CAN ONLY SUSPECT IT IS
REALLY EFFICACY TESTING THAT'S
THE COMPETITION OF IDEAS WHERE
THE BETTER MODEL IS USED BY THE
MORE TECHNOLOGY COMPANIES. THE
AREA SHOOT IN THE FOOT IF YOU
ARE USING IN ADDITION TO WHAT
YOU HAVE TO DO, YOU CAN ONLY
LOSE INNOCENCE AND THEN YOU HAVE
TO FOLLOW-UP AND DELAY WHATEVER
PROCESS. THERE ARE THESE MODELS
SHINE VERY EARLY STAGE OF
INVESTIGATIVE TOXICOLOGY OF
TARGET EVALUATION AND FOR
DIVERTS PHIING ANIMAL STUDIES
YOU WANT TO -- DIVERSIFYING
ANIMAL STUDIES YOU WANT TO SHOW
IT IS NOT RELEVANT SO IT IS
TOXICOLOGY BUT NOT REGULATORY
TOXICOLOGISTS.
>> SO I WOULD JUST MAKE A LITTLE
PUSH BACK HERE FOR IN TERMS OF
DATA SAFETY DATA. OF COURSE THE
VIRUS IS VERY HIGH BUT WE DON'T
HAVE TO USE THEM FOR ALL END
POINTS IF WE HAVE SPECIFIC WELL
DEFINED END POINTS FOR ANIMALS
FULFILLING ALL THE INFORMATION.
BY DOING THAT IT IS NOT THAT WE
ARE SHOWING DEVELOPMENT, IN FACT
THE MORE UNCOMFORTABLE US AS
REGULATORS ARE WITH A PRODUCT
THAT IS GOING TO -- THE SLOWER
THE STUDY WILL BE BECAUSE THE
CLINICAL TRIAL DESIGN WILL BE
IMPACTED, IT WILL HAVE SAFETY
MEASURES, WILL HAVE ONE PATIENT
AT A TIME. SO THERE WOULD BE THE
MORE UNKNOWNS WE HAVE THE MORE
UNCOMFORTABLE WE ARE. THE SLOWER
THE MORE SAFETY MEASURES WE HAVE
TO PUT IN PLACE JUST TO MAKE
SURE THAT THE SAFETY OF CLINICAL
TRIAL PARTICIPANTS IS BEING
ENSURED. SO IF DATA FROM A
SYSTEM VALIDATED, THAT IS WE
KNOW THAT IS BRINGING OUT END
POINT AND OUTCOMES THAT ARE
PREDICTIVE OF HUMAN OUTCOMES
THEN THAT WOULD ACTUALLY HELP TO
THE CLINICAL DEVELOPMENT IN MY
OPINION.
>> I WANT TO TAKE THE LAST
MINUTE COUPLE OF QUESTIONS FOR
EDWIN. THE ONE QUESTION, IT IS A
BEAUTIFUL SYSTEM FOR SCREENING.
WHAT IS THE THROUGH PUT FOR
THAT, HOW MANY COMPOUNDS COULD
YOU SCREEN PER MONTH OR WEEK?
>> I THINK THAT IT IS UP TO
DEFINITELY RIGHT NOW THE MOST WE
HAVE DONE IS BASICALLY -- TEN
PLACE OVER TIME BUT WE CAN SET
UP TO HOW MANY YOU WANT TO DO
ACTUALLY AND THEN RIGHT NOW IT
IS VERY WELL SCALABLE IT IS NOT
REALLY A LIMITATION AT THIS
POINT. WE DO THE ASSAYS HAVE 384
WELL PLATE ACCESS SO HIGHLY
SCALABLE WE HAVE MORE THAN 50
PATIENTS SO FEW ROUNDS OF
SCREENING BUT MOST PART HIGHLY
HIGH THROUGH PUT AND VERY WELL
SCALABLE AS WELL.
>> AND ALWAYS HIGH CONTENT
IMAGING OR DO YOU HAVE FURTHER
READ OUTS?
>> IT IS HIGH CONTENT IMAGING,
CONFOCAL BASED IMAGING ACTUALLY.
IT IS BOTH IN NOT ONLY INFECTION
LEVELS BUT ALSO IN SITU LOOK AT
COMPOUNDS THAT NOT ONLY BENEFITS
IN TERMS OF DEREDUCING VIRAL
LOAD BUT RESCUING PATHOLOGY IN
LUNG TISSUE. WE DO THAT BY USING
THE COMPUTATIONAL BASE AND TO
DEFINE THOSE WE DON'T SEE
MEASURES YET.
>> THAT SHOULD COVER OTHER
QUESTIONS FOR YOU IN THE OPEN
BOX IF YOU WANT TO ANSWER
OFFLINE. WE ARE OUT OF TIME. I
WILL TURN THE MEETING BACK TO
SARINE. THANK YOU, ALL SPEAKERS
THIS HAS BEEN A WONDERFUL
SESSION. THANK YOU FOR THIS
GREAT DISCUSSION AFTERWARDS.
APPRECIATE IT.
>> THANK YOU.
>> THANK YOU SO MUCH, ROBERT RO,
FOR CHAIRING THE WONDERFUL
SESSION AGAIN, I WOULD LIKE TO
THANK AGAIN THE SPEAKERS OF THE
SESSION AND THE WONDERFUL
CONVERSATION AND DISCUSSION THAT
WAS REALLY EXCITING. OKAY. SO WE
ARE AT THE FINAL SESSION. SUPER
EXCITED TO INVITE BACK DR. SIMON
FUNNELL OUR KEYNOTE SPEAKER FROM
THE UK HEALTH SECURITY AGENCY.
DR. NICOLE KLEINSTREUER FROM
ENVIRONMENTAL HEALTH SCIENCES
AND SARA CHERRY FROM UNIVERSITY
OF PENNSYLVANIA, EXCITED TO HEAR
ABOUT DISCUSSIONS AND
PERSPECTIVES ON THE CHALLENGES
AHEAD. THE FLOOR IS YOURS.
>> THANKS. WHAT I DO IS I ASK
YOU FOR YOUR SLIDE FROM THE
FIRST SESSION WHERE YOU
INTRODUCED THE WORKSHOP AND I
JUST WANTED TO SHARE YOUR SLIDE
TO TALK ABOUT WHAT YOU STATED
WERE OBJECTIVES. I THINK WE DID
A GOOD JOB COVERING TODAY. AND
YESTERDAY. YOUR SLIDE.
>> IDE SEE IN PRESENTER MODE.
HAPPY TO SHARE IF YOU WOULD LIKE
TALKING AVAILABILITY OF TISSUE
MODELS AND CHALLENGES IN
BUILDING THEM IN TERMS OF
UTILITY AND LIMITATIONS HOW THEY
CAN BE USED IN DRUG DISCOVERY
DEVELOPMENT WITH THE PROS AND
CONS, WHAT WE THINK GUIDES LINES
AND CONSIDERATIONS MAYBE FOR
DEVELOPING ROBUST REPRODUCIBLE
MODELS. AND THEN TALK ABOUT
AFFORDABILITY ACCESSIBILITY AND
TRANSFERABILITY OF THESE MODELS.
COVER THAT MUCH ABOUT THE
ADVANTAGE AND DISADVANTAGE OF 3D
MODELS OTHER THAN TO PRETTY MUCH
ALL AGREE THERE ARE MAJOR
ADVANTAGES BUT I THINK THERE ARE
LIMITATIONS PERSONALLY AS USER
IT IS DIFFICULT TO ADOPT THESE
AND FIGURE OUT THE DIFFERENCES
ACROSS THE PLATFORMS BUT I
THOUGHT THAT ACCESS AND SIMON
WANTED TO TALK A LITTLE BIT
ABOUT SOME ASPECTS OF THIS SO
I'M GOING TO TURN IT OVER TO HIM
NOW.
>> THANK YOU, ALSO THANK YOU TO
NSP CORE GROUP WHO INTRODUCED ME
TO THIS DAZZLING ARRAY OF TALKS
THE LAST TWO DAYS. SESSION 1, 2
AND 3 TOGETHER FORM WHAT I TRIED
TO DESCRIBE AS A SCIENCE, THAT
TRAIN HAS LEFT THE STATION AND
I'M PLEASED TO BE ON THE LAST
CARRIAGE. WHEN WE TALK
REGULATORY ACCEPTANCE OF THE
DATA BEING GENERATED, WHEN WE
SAW SLIDE OF REQUIREMENTS AND
THE WHETHER THE POINT SHE PUT
YOU WANT -- BULLET POINTS SHE
PUT UP WE CAN TAKE AWAY FROM THE
BULLET POINTS. MY PERCEPTION IS
SARS COV-2 AS TRAGIC AS IT HAS
BEEN FOR THE PLANET, IT HAS
ACTUALLY BEEN A GIFT FOR THIS
TYPE OF TECHNOLOGY. WHEN WE
REFLECT HOW ANIMAL MODELS HAVE
DELIVERED AND ARGUABLY THE
HAMSTER HAS BEEN VERY HELPFUL
BECAUSE THERE IS NOT ENOUGH
NON-HUMAN PRIMATES WITH DATA
THAT WE NEED WHEN WE THINK ABOUT
OMICRON AND BA 2, BA 4, BA 5
CURRENT CIRCULATING STRENGTH,
THESE VIRUSES ARE DIFFERENT THAN
THE PROTOTYPE VIRUS. WE CAN'T --
ANYBODY TRIES TO GROW BA 2 BA 4
OR 5 THEY STRUGGLE WITH THAT.
THEY ALWAYS USE STEM CELL WHICH
HAS A PLASMA WITHIN IT, WHICH
HAS (INAUDIBLE) AND WHENEVER WE
GROW VIRUS IN THAT CELL LINE
THEY TEND TO BE TRUE TO THE
ISOLATES. BUT I KNOW OTHERS HAVE
USED OTHER CELL LINES BUT
VARIOUS HAVE BEEN A GIFT TO US
BECAUSE WE HAVE BEEN ABLE GROW
OMICRON IN THOSE CELLS IN A WAY
WE COULDN'T DO SO BUT AT THE END
OF THE DAY IT IS STILL A CELL,
AT HEART OF THE ISSUE OF
ANTIVIRAL DRUG DEVELOPMENT IS
GETTING THE RIGHT VIRUS TO START
WITH. YOU CAN HAVE THE BEST
SYSTEM ON THE PLANET IF YOU PUT
IN A VIRUS, ANY WAY COMPROMISE
FROM THE ACTUAL TARGETS YOU HAS
-- YOU WERE AT BAD START. WHEN
WE THINK ABOUT OMICRON IT IS
ALSO REI CAN LOUSE TO SHOW ANY
CLINICAL SIZE AT ALL UNLIKE
HUMAN YOU WILL SEE A NUMBER OF
CLINICAL SITES, SO WHEN WE THINK
COMPARING TO THE GOLD STANDARD,
WHICH IS ONE BULLET POINT ON THE
SLIDE, THEY MAY BE A GIFT. SO
STRAIGHT AWAY YOU WANTED TO --
THEY KNOW THAT AND THEY HAVE
GREAT APPETITE TO ACCEPT THIS
KIND OF DATA AND THE OTHER GIFT
WAS THE PAPER I DESCRIBED,
YESTERDAY AND REFERRED TO AGAIN
TODAY ABOUT HYDROXYCHLOROQUINE
AND HEMATONIB, THESE DRUGS ARE
DIFFICULT COURSES TO THE
COMMUNITY BECAUSE THEY
CONSOLIDATE MPS AS A MEDICAL
TOOL IN THE QUEST FOR ANTIVIRALS
AND WHEN I THINK ABOUT WHAT SARA
HAS PRESENTED, WHAT CALVIN
PRESENTED, WHAT -- THERE'S SO
MANY TALKS HERE, THAT GIVES ME
INSPIRATION THAT YOU CAN VERY
QUICKLY PUTTING TO A REGULATORY
PACKAGE ACCEPTABLE BY THE FDA
BASED ON THE DEFICIENCIES OF THE
CURRENT GOLD STANDARD WHICH MAY
NO LONGER WORK FOR SOME OF THESE
VARIANTS. I DON'T WANT TO TAKE
UP ALL 20 MINUTES BUT SAW
WONDERFUL QUOTATIONS IN SOME OF
THE SLIDES, ONE FROM EINSTEIN
SAYING WITHIN ANY CRISIS THERE'S
OPPORTUNITIES WHERE WE ARE THERE
NOW WITH SARS COV-2 AND I SAW
ANOTHER QUOTE SAYING THE
DIFFICULTY OF FUTURE TECHNOLOGY
IS NOT ACCEPTANCE OF THE NEW BUT
THE RESISTANCE TO REJECT THE
OLD. THAT IS ONE OF THE -- THAT
I DO THINK THERE'S -- WITHIN THE
FDA TO UNDERSTAND THESE ISSUES
SO I THINK THAT I WILL STOP
THERE AND ALLOW SARA AND NICOLE
TO TALK IN THE MINUTES THAT
REMAIN IN THIS DISCUSSION.
>> THANKS, SIMON. I WILL WEIGH
IN AS FAR AS MY 30,000-FOOT VIEW
PERSPECTIVE FROM THE WORK THAT
THE EXCELLENT TALKS WE HAVE
HEARD THE LAST DAY AND A HALF.
AND WHERE THIS FIELD IS GOING
AND SPECIFICALLY FROM
PERSPECTIVE OF STARTING THE
QUALIFY SYSTEMS FOR SPECIFIC
CONTEXT AND USE AND GAINS MORE
ACCEPTANCE AS I HAVE MENTIONED I
FULLY AGREE THAT WHILE
INCREDIBLY TRAGIC THE COVID
PANDEMIC HAS PROVIDED JUST AN
OUTSTANDING OPPORTUNITY TO
REALLY STRESS TEST THESE SYSTEMS
AND SHOW HOW USEFUL THEY ARE AND
HOW IMPORTANT THEY ARE, IN
PROVIDING US HUMAN RELEVANT
INSIGHT INTO THE PATHOPHYSIOLOGY
AND DISEASE MECHANISMS PLATFORMS
FOR THERAPEUTIC TREATMENT BUT
STEPPING BACK THINKING MORE
GENERALLY ABOUT MPS AND HOW WE
GET TO REGULATORY ACCEPTANCE AND
ABILITY TO TRULY HAVE THAT
SCIENTIFIC CONFIDENCE TO RELY ON
THE DATA THOSE SYSTEMS ARE
PROVIDING, I THINK THERE IS BEEN
MANY INSTANCES OF MEETINGS VERY
USEFUL MEETINGS WHERE DEVELOPERS
AND END USERS IN PHARMA DRUG
DEVELOPMENT SPACE AND REGULATORS
COME TOGETHER AND DISCUSS
QUALIFICATION CRITERIA AND
REGULATORY NEEDS, AROUND
COMPARISON THE EXISTING DATA AND
SPECIFIC END POINTS BUT THERE IS
NO CONSENSUS TO WHAT THAT LOOKS
LIKE SO I THINK WE ARE MOVING
TOWARD THAT SPACE. AS WE ENGAGE
MORE AND MORE WITH REGULATORY
AUTHORITY IN THE U.S. IN EUROPE
IN ASIA. AND DEVELOP
QUALIFICATION CRITERIA, THE
IQMPS AFFILIATE IS ONE OF THE
WORLDWIDE LEADERS IN THIS SPACE,
THERE IS A NICE EXAMPLE
PUBLISHED ON LIVER CHIP,
QUALIFICATION CRITERIA AND
STANDARD COMPOUND SETS FROM MY
PERSPECTIVE HELPING TO LEAD
VALIDATION ORGANIZATION
REFERENCE COMPOUNDS ARE PRETTY
CRITICAL IN TERMS OF BEING ABLE
TO DEMONSTRATE THE SYSTEM IS
REPRODUCELY ROBUSTLY GIVING YOU
THE INFORMATION YOU EXPECT SO
YOU CAN HAVE CONFIDENCE WHEN YOU
ARE TESTING SOMETHING NEW AND
UNKNOWN THAT YOU CAN TRUST THAT
DATA AN INTERPRET THOSE RESULTS
SO I THINK THAT THERE -- WE
REALLY NEED TO HAVE MORE GLOBAL
INTERACTIONS AMONG DIFFERENT
REGULATORY AUTHORITIES AND
METHOD DEVELOPERS TO ESTABLISH
STANDARD COMPOUNDS, THOSE
QUALIFICATION CRITERIA THAT ARE
TARGET ORGANS SPECIFIC. SO THAT
IS ONE THING TO SEE MORE
EMPHASIS PUT ON. IT IS NOT
ADEQUATELY RECOGNIZED HOW MUCH
OPPORTUNITY FROM THERE IS FOR
COMPANIES AND TECHNOLOGY
DEVELOPERS TO EBB GAUGE WITH THE
FDA THROUGH THE ICAM PROGRAM FOR
SURE BUT REACHING OUT TO THE FOR
EXAM FELONY THE ALTERNATIVE
METHODS WORK -- EXAMPLE, THE
ALTERNATIVE METHODS WORKING
GROUP AT THE FDA AND STARTING A
DIALOGUE. THERE ARE MANY CENTERS
NOT JUST CBER BUT CDER VERY
ENGAGED IN THE SPACE AND VERY
INTERESTED IN GETTING
SUBMISSIONS OF DATA RELYING ON
MPS, THAT ARE TARGETED FOR
SPECIFIC CONTEXT OF CRUISE BUT
IF I HAVE LEARNED ANYTHING IN
THE LAST TEN YEARS WITH NICEATM
IT IS THE ABSOLUTE IMPORTANCE OF
ENGAGING WITH YOUR END USERS
PARTICULARLY REGULATORY DECISION
MAKERS FROM DAY ONE, HAVING THAT
BE A CONSTANT ITERATIVE CYCLE OF
COMMUNICATION SO THAT YOU ARE
DEVELOPING METHODS TARGETED
TOWARDS THEIR NEEDS AND YOU ARE
UNDERSTANDING THEIR DECISION
CONTEXT AS GOING THROUGH THE
PROCESS OF DEVELOPMENT AND
QUALIFICATION AND ESTABLISHING
CONFIDENCE IN THOSE NEW
APPROACH. BUT I THINK THE SAME
IS ALSO TRUE WITH I MENTION WE
HAD A MEETING WITH EUROPEAN
MEDICINE AGENCY, THEY HAVE THE
INNOVATION TASK FORCE, SO THEY
HAVE SAFETY WORKING PARTY AND
THE INNOVATION TASK FORCE AND
THIS SAFE HARBOR PROGRAM WHERE
THERE IS GUIDANCE ON HOW THESE
DIFFERENT ROUTES OF
COMMUNICATION CAN BE -- WHERE
YOU CAN GET REGULATORY ADVICE ON
THE UTILITY AND VALIDITY OF
MODELS AND DATA GENERATED AND
COMPARE ALONGSIDE DATA GENERATED
IN TRADITIONAL STUDIES. SO
THERE IS GUIDANCE HOW TO ACCESS
THOSE ROUTES BUT WHAT WE HEAR IS
THE UPTAKE IS VERY LOW SO THERE
IS FACTORS THAT CONTRIBUTE TO
THAT, LACK OF AWARENESS AROUND
OPPORTUNITIES, CONCERNS
MISPLACED OR NOT THAT IF
DISCUSSIONS ARE NOT FAVORABLE
THAT MIGHT COUNT AGAINST THEM
BUT HELPING SOCIALIZE
DISSEMINATE THAT INFORMATION
ABOUT OPPORTUNITIES TO ENGAGE
WITH REGULATORY AUTHORITIES, AND
GET DIALOGUE GOING AND GET THAT
ADVICE, IN A SAFE HARBOR WAY IS
HELPFUL IN MOVING THE FIELD
FORWARD. THAT IS MY PIECE, I
WILL STOP THERE.
>> I THINK THAT THAT IS RIGHT,
FROM THE END STAGE, REGULATORY
AGENCIES IS REALLY IMPORTANT BUT
I ALSO THINK THAT IT IS HELPFUL
TO ENGAGE MORE OF THE USER BASE
TO THIS COMMUNITY AND HAVE MORE
ACCESSIBILITY TO YOUR
(INAUDIBLE) FOR EXAMPLE, THE
ONLY WAY I START TO UTILIZE
THESE MODEL IS COLLABORATE WITH
NCATS AND WITH GATES FOUNDATION
WHO FACILITATED THE ABILITY NOT
ONLY TO ACCESS MODEL BUT LEARN
HOW TO USE THEM. SO I REALLY
THINK THAT THAT IS A REAL GAP,
IT IS CHALLENGING FOR
VIROLOGISTS TO ACCESS THEM SO
THE VIROLOGISTS BRING DRUGS
FORWARD, NOT ACTIVE BECAUSE THEY
ARE USING FLAWED MODELS SO IF WE
CAN STEP BACK AND BRING SOME OF
THESE MODELS MORE EASILY TO
REGULAR SCIENTISTS THEN I THINK
WE WOULD BE BRINGING FORWARD
MORE DRUGS THAT WOULD ULTIMATELY
BE ACTIVE, HOPING TO.
COUP WITH STRATEGIES. TO MAKE
ACCESSIBLE AND UNDERSTANDABLE
WITH MORE TO THE STANDARDIZATION
TO MAKE IT EASIER.
>> YESTERDAY SOMEONE ASKED ME A
QUESTION WHAT IMMUNE CELLS ARE
INTRODUCED INTO
MICROPHYSIOLOGICAL SYSTEMS AND
TODAY WE HEARD ABOUT THAT. AND
EFFICACY AS WELL. WE HEARD ABOUT
NUTRIENTS BEING INTRODUCED,
INTRODUCE WHOLE BLOOD INTO THE
CHANNEL WHICH IS A COMPLEX SET
FOR PERIPHERAL BLOOD AND WE
HEARD FROM CALVIN ABOUT TISSUE
RESIDENT T-CELLS, THAT IS A HOT
TOPIC BECAUSE THAT IS DIFFICULT
TO MEASURE IN THE FIELD. I WANT
TO REFLECT ON THE GOLD -- OLD
WAY OF DOING THINGS. THE OLD WAY
IS PERHAPS TO USE CELL LINES
MAYBE INSECTS AND THEN MOVE UP
TO SMALL ANIMALS LIKE MICE AN
RATS HAMSTERS AND THEN ENTER NHP
FOR FINAL DATA SET AND FOR
SELECT AGENTS. MAYBE THE SORT
SYSTEMS SARA AND THE AMAZING
LAST SPEAKER WAS TALKING ABOUT
IN TERMS OF SCREENING THOUSANDS
OF DRUGS IS A VERY USEFUL
PRELIMINARY SCREENING MODE THEN
WHEN WE GET INTO PHASE 1 STUDY
WE CAN LEVERAGE PHASE 1 STUDIES
IN A WAY THAT I HADN'T THOUGHT
OF AND THAT IS TO TAKE A PHASE 1
STUDY TAKE BLOOD BUT ALSO TAKE A
SMALL BRUSH OR BIOPSY TO SEE
TISSUE BIOLOGY I DON'T KNOW SHY
THAT INCLUDES T-CELL -- RESIDENT
T-CELLS, SO MAYBE TOGETHER THAT
IS FORMIDABLE DOUBLE ACT INITIAL
SCREENING AND THEN ACTUAL PHASE
1 STUDY. MAYBE THE -- THIS
CONFERENCE IS ABOUT SPECIFICALLY
ANTIVIRALS AND WE HAVE
INEVITABLY ANTIVIRALS WILL
INCLUDE DRUGS WHICH STIMULATE
THE IMMUNE SYSTEM OR ACT AS IF
THEY ARE PART OF IT BUT MAYBE
THE LOW HANGING FRUIT IS A DRUG
WHICH DOES WORK IN HUMANSES BUT
JUST CAN'T WORK IN AN ANIMAL. SO
YOU ARE A WINNER THERE BECAUSE
THERE IS NOT GOLD STANDARD.
THIS IS WHAT WE HAVE AND THEN
YOU CAN SAY YOU CAN RELEVERAGE
HYDROXYCHLOROQUINE FORRY IN THAT
SOMETHING LIEU THE REGULATORY
PATHWAY AND BUILD ON IT WHICH IS
MORE COMPLICATED THAN WAS TRYING
TO ARTICULATE YESTERDAY BUT THE
DIVERSITY MODELS I HEARD ABOUT
TODAY AND OBVIOUSLY REALLY
REGRET NOT HAVING BEEN ABLE TO
ATTEND THE CONFERENCE COUPLE OF
WEEKS AGO BECAUSE CLEARLY THERE
ARE MANY SOLUTIONS IN THE MST
CORE HUB WHICH WILL ANSWER
QUESTIONS THAT WILL EVOLVE AS WE
HIT VARIOUS REGULATORY
CHALLENGES. AND I DON'T SAY THE
HURDLES, I SAY THEY ARE
CHALLENGES. I THINK WE WILL
MAKE MISTAKES ALONG THE WAY BUT
AS I SAID BEFORE IN ANSWERING
QUESTIONS FROM YESTERDAY A
MISTAKE OR FAILURE IS AN
OPPORTUNITIES TO IMPROVE. SO
LET'S NOT BE DOWN HEARTED ABOUT
FAILURE BECAUSE IT IS THE SPACE
TO MOVE FORWARD. I CAN SEE
SUCCESS IN WHAT I HAVE HEARD
ABOUT ESPECIALLY SESSION 2 AND 3
TODAY, MIND BOGGLING AND SO
HAPPY TO HAVE BEEN INVITED TO
PRESENT AND ATTEND SO I WILL
STOP THERE.
>> ALL RIGHT, THANK YOU VERY
MUCH SARA, NICOLE, SIMON FOR THE
WONDERFUL WRAP UP OF THIS REALLY
EXCITING TWO DAYS. I HAVE ONE
FINAL SLIDE TO SHARE. IT IS A
THANK YOU SLIDE. SO I WOULD
LIKE TO WRAP UP TODAY BY TAKING
EVERYONE THE CHAIRS THE
ORGANIZER, THE SPEAKERS, F
WONDERFUL TWO DAYS, THANKS
ATTENDEES AND THANK FOR EVERYONE
FOR ATTENDING. HOPEFULLY YOU
LEARNED A LOT AND THIS WAS
REALLY BENEFICIAL TO YOU AND FOR
YOUR RESEARCH. WE DO LIKE TO GET
YOUR FEEDBACK SO I KNOW ABIGAIL
MESSAGED ALREADY, SHE WILL REACH
OUT TO EVERYONE WITH THE
FEEDBACK LINK, I BELIEVE IT IS
IN THE CHAT, SHE ALREADY SENT IT
TO YOU ALL AND SHE WILL ALSO
EMAIL EVERYONE WITH THE FEEDBACK
SO PLEASE PROVIDE FEEDBACK WITH
LOTS OF -- WHAT YOU THINK WE DID
RIGHT AND WHERE WE CAN IMPROVE.
I ALSO INVITE EVERYONE TO
CONNECT WITH US SO WITH THE --
WITH OUR PROGRAM THE ASSAY
GUIDANCE MANUAL PROGRAM AND
HOPEFULLY ATTEND FUTURE
WORKSHOPS WE RUN, SO AGAIN ON
BEHALF OF THE ASSAY GUIDANCE
MANUAL PROGRAM, THANK YOU, VERY
MUCH FOR ATTENDING FOR SPEAKING
AND WITH THIS I WOULD LIKE TO
ADJOURN TODAY. YOU ALL HAVE A
WONDERFUL EVENING. THANK YOU.