>>WELCOME, EVERYONE, TO THE
ASSAY GUIDANCE WORKSHOP ON 3D
TISSUE MODELS FOR ANTIVIRAL DRUG
DEVELOPMENT.
I AM THE EDITOR-IN-CHIEF OF THE
ASSAY GUIDANCE MANUAL AND THE
LEAD OF THE ASSAY GUIDANCE
MANUAL PROGRAM HERE AT THE
NATIONAL CENTER FOR ADVANCING
TRANSLATIONAL SCIENCES.
AT THE NATIONAL INSTITUTES OF
HEALTH.
WE ARE EXTREMELY EXCITED TO HAVE
YOU ALL JOIN US TODAY IN THIS
EXCITING WORKSHOP THAT WILL RUN
FOR TWO DAYS, TODAY, JUNE 7, AND
TOMORROW, JUNE 8 AS WELL.
SO THIS WORKSHOP WAS ORGANIZED
BY THE NATIONAL CENTER FOR
ADVANCING TRANSLATIONAL SCIENCES
AS WELL AS THE BILL AND MELINDA
GATES FOUNDATION, AS WELL AS THE
NATIONAL INSTITUTES OF ALLERGY
AND INFECTIOUS DISEASES.
AND WE'RE GOING TO START THE
PROGRAM TODAY BY INTRODUCTORY
COMMENTS BY LEADERSHIP FROM
THESE THREE INSTITUTES.
WE'LL START WITH DR. JONI
RUTTER, WHO'S THE ACTING
DIRECTOR OF THE NATIONAL CENTER
FOR ADVANCING TRANSLATIONAL
SCIENCES AT THE NATIONAL
INSTITUTES OF HEALTH.
WE'RE REALLY EXCITED TO HAVE YOU
HERE, JONI.
WELCOME.
THANK YOU SO MUCH FOR BEING
HERE.
WE'RE LOOKING FORWARD TO HEARING
YOUR COMMENTS.
THE FLOOR IS YOURS.
>> OKAY.
THANK YOU SO MUCH, IT'S REALLY A
PLEASURE TO BE HERE.
HELLO, EVERYONE, AND WELCOME TO
THE MEETING.
AGAIN, MY NAME IS JONI RUTTER,
ACTING DIRECTOR OF NCATS.
BEFORE I BEGIN, I WANT TO THANK
SEVERAL OF OUR PARTNERS FOR THE
FORESIGHT AND VISION FOR THIS
MEETING.
AND THOSE INCLUDE
REPRESENTATIVES FROM EACH OF
THOSE ORGANIZATIONS THAT YOU SEE
LISTED THERE ON THAT SLIDE.
RUDOLPHO, MINDY, ANNE, TIM AND
MARK ALL FROM NIAID WHO'VE
REALLY HELPED FROM THEIR
PERSPECTIVE BRING IMPORTANT
CONTRIBUTIONS TO THIS MEETING.
I'D LIKE TO ALSO THANK TIM
DUNCAN AND ROBERT JORDAN FROM
THE BILL AND MELINDA GATES
FOUNDATION FOR THEIR SUPPORT AS
WELL AND THEN ALSO CAL, MARK,
STEPHANIE, MATT, EMILY, ALL FROM
NCATS, CONTRIBUTING THEIR
PERSPECTIVES, AND THEN
ESPECIALLY AS YOU'VE HEARD
SERENE AND ABIGAIL WHO HAVE BEEN
VERY MUCH HELPING US ORK STATE
THIS ENTIRE EFFORT.
SO THANK YOU TO ESPECIALLY
ABIGAIL AND SERENE FOR PULLING
THIS MEETING TOGETHER AND ALL
THE OTHER PARTNERS AND
CONTRIBUTORS WHO HAVE HELPED
BRING THIS MEETING AND THE
AGENDA WHICH IS INCREDIBLY
FANTASTIC TO THE FORE HERE.
THIS MEETING SEEKS TO GUIDE THE
DEVELOPMENT OF STANDARDIZED 3D
CELLULAR ASSAYS WITH THE HOPE OF
HELPING THE COMMUNITY TO
SUCCESSFULLY AND MORE
EFFICIENTLY DEVELOP, TEST,
VALIDATE AND DEPLOY THERAPEUTICS
FOR FUTURE PANDEMIC THREATS.
AND THEREFORE ALSO PROVIDE PROOF
OF CONCEPT FOR THESE MODELS THAT
CAN DO THE SAME FOR OTHER PUBLIC
HEALTH MATTERS.
AND RIGHT NOW, THESE 3D MODELS
ARE SOPHISTICATED TOOLS IN THE
TOOLBOX, BUT CAN THEY BE MORE?
CAN THEY BE PARADIGM-SHIFTING IN
A SPACE OF THERAPEUTICS
DEVELOPMENT THAT IS IN DIRE NEED
OF TRANSFORMATION?
SO OUR MISSION AT NCATS IS TO
CATALYZE THE GENERATION OF
INNOVATIVE METHODS AND
TECHNOLOGIES THAT WILL ENHANCE
THE DEVELOPMENT, TESTING AND
IMPLEMENTATION OF PLATFORM
DIAGNOSTICS AND THERAPEUTICS
ACROSS A WIDE RANGE OF HUMAN
DISEASES AND CONDITIONS, AND
ALWAYS WITH AN EYE TOWARDS
TURNING PROMISING RESEARCH
DISCOVERIES INTO HEALTH
SOLUTIONS.
AND OUR STRATEGY AT NCATS IS
TWOFOLD.
EVERY PROJECT WE TAKE ON
ADVANCES THE SCIENCE OF THE
PARTICULAR PROJECT AT HAND, BUT
IT ALSO SEEKS TO IDENTIFY AND
ADDRESS COSTLY AND
TIME-CONSUMING BOTTLENECKS THAT
SLOW DOWN OR PREVENT THE
TRANSLATION OF RESEARCH.
AND SO WE SEEK TO INCREASE THE
PACE OF RESEARCH BY TESTING AND
DEVELOPING THERAPEUTIC SO THAT
WE CAN ADVANCE ON THE SCALE OF
MANY DISEASES AT A TIME, NOT
JUST ONE DISEASE AT A TIME.
AND OUR ULTIMATE DRIVING HOPE IS
TO GET MORE TREATMENTS TO ALL
PEOPLE MORE QUICKLY.
AND LOTS OF TIMES THAT'S REALLY
THROUGH ENABLING AND DE-RISKING
PLATFORM-BASED TECHNOLOGIES FOR
THERAPEUTIC DEVELOPMENT.
LIKE 3D TISSUE MODELS THAT WE'RE
TALKING ABOUT OVER THE NEXT
COUPLE OF DAYS.
SO IMAGINE IF THE 3D MODELS ARE
SUCCESSFUL AT INCREASING OUR
ABILITY TO ADDRESS TOXICOLOGICAL
ISSUES OR EFFICACY PREDICTIONS,
FOR EXAMPLE, THEN WE CAN HASTEN
NEEDED NEW DRUG DEVELOPMENT
APPROACHES THAT WOULD OTHERWISE
TAKE DECADES.
OR WE COULD ENSURE THAT THERE
ARE PROVEN OFF THE SHELF HUMAN
CELL-BASED MODELS FOR DRUG
REPURPOSING EFFORTS IN TIME OF
URGENCY WHERE AN ANIMAL MODEL,
FOR EXAMPLE, MAY NOT BE
ULTIMATELY AS HELPFUL AS HUMAN
CELL-BASED MODELS OR WHERE THEY
MAY BE A LITTLE BIT MORE
LIMITING BECAUSE OF MULTIPLE
CO-MORBIDITY TYPES OF
CONDITIONS.
SO THESE ARE THE IMPORTANT KINDS
OF THINGS WE'LL BE TALKING ABOUT
OVER THE NEXT COUPLE OF DAYS.
NOW IN 2019, THERE WAS A DRUG
DISCOVERY TODAY ARTICLE THAT
PURPORTED THE POTENTIAL OF 3D
MODELS COULD REDUCE R & D DRUG
COSTS BY UPWARDS OF 26%.
WHERE IMPROVEMENTS IN CLINICAL
TRIAL SUCCESS PREDICTION RATES
WOULD REALLY BE THE MAIN DRIVERS
OF WHERE THOSE ANTICIPATED COSTS
COULD FIND -- WE COULD FIND COST
SAVINGS.
AND WE KNOW ABOUT 85 TO 90% OF
DRUG CANDIDATE ARE -- THEY FAIL
BECAUSE OF DRUG EFFICACY
CONCERNS OR SAFETY CONCERNS, AND
THAT'S WHERE THOSE COST SAVINGS
ARE LURKING, AND IF THAT'S THE
MAJOR DRIVE OF R & D COSTS, THEN
BETTER PREDICTIVE MODELS COULD
HELP CHIP AWAY AT THOSE COSTS SO
WE NEED TO FIND WAYS TO GET THEM
INCULCATED INTO THE
TRANSLATIONAL DRUG DEVELOPMENT
IN THE GENERAL ECOSYSTEM FOR
DRUG DEVELOPMENT.
IN ADDITION TO TALKING ABOUT
2019 IN THAT ARTICLE, THEY ALSO
SUGGESTED THE TECHNOLOGY WILL
HELP TO MAKE QUICKER AND MORE
PRECISE DECISIONS DURING THESE
PRE-CLINICAL STAGES OF DRUG
DEVELOPMENT.
SO THE PROMISE OF 3D MODELS IS
REALLY -- THERE IS STILL A LONG
ROAD AHEAD TO UNDERSTAND WHETHER
THAT PROMISE WILL BECOME A
REALITY AND THIS MEETING NEEDS
TO HELP FOCUS AND SHAPE THE PATH
FORWARD IN THE CRITICAL AREA OF
ANTIVIRAL DRUG DEVELOPMENT.
IN TERMS OF REGULATORY ADOPTION,
THERE ARE BIG QUALIFICATION
CHALLENGES THAT 3D MODELS NEED
TO MEET BEFORE WIDER ADOPTION
WILL ULTIMATELY BE SEEN.
WE NEED TO GET HIGHER HUMAN
RELEVANCE AND DISEASE RELEVANCE
EARLIER IN THE DRUG DEVELOPMENT
PIPELINE FOR THOSE TO REALLY
MANIFEST AND TAKE EFFECT, AND
ANTIVIRAL DRUG DEVELOPMENT IS A
RIPE AREA FOR THIS TO BE DONE SO
THERE'S NO TIME LIKE THE PRESENT
AND CERTAINLY THIS PARTICULAR
MEETING WILL HELP TO ADDRESS
THAT.
AND I WANT TO HIGHLIGHT THREE
KEY AREAS THAT I HOPE WILL BE
DISCUSSED DURING OUR NEXT TWO
DAYS TOGETHER.
FIRST ARE SOME HIGH LEEFL
OUTSTANDING QUESTIONS AND BIG
CHALLENGES THAT ARE STILL AT
PLAY.
WE NEED TO ESTABLISH AND BEGIN
TO QUANTIFY THE CLINICAL
PREDICTABILITY OF 3D TISSUE
MODELS.
HOW GOOD ARE THEY IN
RECAPITULATING DISEASES OF
INTEREST?
WHAT IS THE PHYSIOLOGICAL
COMPLEXITY NEEDED IN A TISSUE
MODEL TO BEST PREDICT THE
CLINICAL RESPONSES?
HOW IMPORTANT ARE THE IMMUNE
COMPONENTS OR ENDOTHELIAL
COMPONENTS IN THESE MODELS?
AND ASSESSING WHETHER THE TISSUE
HAS THE APPROPRIATE SET OF
BIOMARKERS, WHETHER THOSE
BIOMARKERS BE FOR PREDICTIVE
PURPOSES OR SAFETY PURPOSES OR
MONITORING PURPOSES.
ASSESSING THOSE BIOMARKERS FOR
EVALUATING AND ACCURATELY
DISTINGUISHING NORMAL AND
DISEASE STATES AND HOW THOSE
BIOMARKERS CAN BE USED IN TERMS
OF CONTEXT OF USE FOR THOSE 3D
MODELS IS GOING TO BE IMPORTANT.
WHAT'S THE BIOLOGY AND THE
PATHOLOGY THAT THESE MODELS WILL
REPORT ON.
AND THEN VALIDATING THAT THEY
CAN PREDICT PHARMACOLOGICAL
RESPONSES OR THERAPEUTICS IN
HUMANS FROM BOTH A SAFETY AND
TOX STANDPOINT AS WELL AS AN
EFFICACY STANDPOINT, THESE ARE
ALL OPEN QUESTIONS THAT ARE
LIKELY TO BE MODEL AND
QUESTION-DEPENDENT.
SO WE NEED TO DEVELOP MULTIPLE
MODELS TO REFLECT A VARIETY OF
DIFFERENT, FOR EXAMPLE,
CARDIOVASCULAR OR HEPATIC OR
NEURONAL OR IMMUNE TOXICITIES,
FOR EXAMPLE, OR FOCUS ON AREAS
WHERE NO PHYSIOLOGICALLY OR
PHARMACOLOGICALLY RELEVANT
MODELS ARE AVAILABLE.
AND THESE ARE AREAS SUCH AS RARE
DISEASES OR RARE CANCERS OR EVEN
IP FORECASTTIOUS EVEN
INFECTIOUS DISEASES WE'LL BE
TALKING ABOUT.
THE SECOND ISSUE IS WITH THESE
MAJOR GAPS STILL IN THE FIELD,
HOW CAN WE IMPROVE UPON THEM?
SO TECHNOLOGICAL ROBUSTNESS AND
REPRODUCIBILITY FOR SCREENING
ASSAYS IS ONE OF THE KEY
CHALLENGES.
AND AS WE ACHIEVE HIGHER
COMPLEXITY IN OUR 3D MODELS, HOW
CAN WE REGAIN GROUND IN
IDENTIFYING A SORT OF MEDIUM TO
HIGH THROUGHPUT ASSAYS FOR
SCREENING FOR THESE TYPES OF 3D
MODELS.
AS YOU INCREASE COMPLEXITY IN
THE 3D MODELS, YOU TEND TO LOSE
THAT HIGH-THROUGHPUT APPROACHES
FOR ASSAYS, SO HOW CAN WE REGAIN
SOME OF THAT MEDIUM OR
HIGH-THROUGHPUT.
HOW CAN WE PERSONALIZE TISSUE
MODELS SO THAT WE CAN ENSURE
THAT MODELS REPRESENT DISEASE
AND POPULATION HETEROGENEITY TO
BETION ENSURE
BEST ENSURE THE RESULTS ARE
GENERALIZABLE.
THEN HOW DO WE INTEGRATE THOSE
TISSUE MODELS EARLIER IN THE
DRUG DISCOVERY PIPELINE?
THIS IS HUGELY CRITICAL.
I THINK ONE OF THE KEY ASPECTS
OF THIS IS UNDERSTANDING HOW
HUMAN RELEVANCE OF TOXICITY IN
ANIMAL STUDIES, BY DOING
COMPARATIVE MEDICINE RESEARCH IN
3D ANIMAL MODELS COMPARED TO 3D
HUMAN MODELS OF KNOWN COMPOUNDS,
FOR EXAMPLE, THAT DIDN'T SHOW
THAT -- THAT SHOWED FAILURE IN
THE HUMAN CONTEXT BUT ACTUALLY
DIDN'T SHOW FAILURE IN THE
ANIMAL CONTEXT, THAT'S GOING TO
BE IMPORTANT TO GO
RETROSPECTIVELY TO UNDERSTAND
WHY THAT IS AND IN WHAT AREAS
THOSE TEND TO HAPPEN.
SO THOSE KINDS OF RETROSPECTIVE
EXPERIMENTS ARE GOING TO BE
IMPORTANT FOR GAINING CONFIDENCE
IN HUMAN CELL-BASED MODELS GOING
FORWARD.
AND THEN WORKING WITH REGULATORY
AGENCIES TO DEVELOP GUIDELINES,
WHAT NEEDS TO BE DONE FOR THESE
MODELS TO BE BROADLY ADAPTED BY
PHARMA AND BY REGULATORY
AGENCIES FOR IND FILING, WHAT'S
GOING TO BE NEEDED FOR THE
REGULATORY PACKAGES SUBMITTED.
THESE ARE OTHER ISSUES THAT NEED
TO BE UNDERSTOOD.
AND THEN OF COURSE BACK TO
BIOMARKERS, DEVELOPING THE
QUANTITATIVE ASSAYS THAT
BIOMARKERS CAN HELP SUPPORT FOR
CLINICAL END POINTS IN 3D MODELS
ARE ANOTHER, OF COURSE, KEY
AREA.
LASTLY, WHAT ARE THE BIGGEST
OPPORTUNITIES FOR THE ANTIVIRAL
DRUG DEVELOPMENT SPACE, AND
THAT'S GOING TO BE THE FOCUS OF
THIS CONVERSATION OVER THE NEXT
COUPLE OF DAYS, BUT HOW CAN 3D
MODELS HELP US PREPARE BETTER
FOR THE CURRENT AND THE NEXT
PANDEMIC.
THAT'S A KEY QUESTION.
SO THERE ARE MANY SCIENTIFIC
TECHNOLOGICAL AND REGULATORY
CHALLENGES THAT STILL REMAIN,
BUT THE 3D MODEL AND THE FIELD
THAT IS REALLY SORT OF COME ON
THE SCENE IN FULL FORCE HAS
SHOWN REMARKABLE
PREDIBILITIABILITY DURING COVID
AND THE 3D MODELS HAVE A BIG
PLACE IN ADVANCING OUR DRUG
DEVELOPMENT GAIN.
SO THE UTION VISION FOR T
DRIVE UTILITY AND ADOPTION OF
THESE 3D MODELS, WORK TOWARDS
THE HARMONIZATION OF REGULATORY
USE AND STANDARDIZATION OF
PLATFORMS AND THEN CHART A PATH
FORWARD THAT WILL EP ABLE US O
BETTER BE PREPARED FOR TREATING
NOT JUST THE CURRENT COVID
PANDEMIC, BUT TO BE MORE
PREPARED FOR ANY FUTURE PUBLIC
HEALTH CRISES.
SO WITH THAT, I WANT TO THANK
AGAIN ALL OF THE SPEAKERS FOR
JOINING US TODAY.
IT'S A LITANY OF REALLY BIG
QUESTIONS AND BIG TOPICS TO
DISCUSS, BUT I KNOW THAT THIS
GROUP WILL PULL TOGETHER AND
IDENTIFY A REALLY GOOD PATH
FORWARD.
SO I LOOK FORWARD TO A VERY
PRODUCTIVE MEETING, AND WITH
THAT, SARINE, I'LL TURN IT BACK
TO YOU TO USHER FORWARD TO THE
NEXT SPEAKERS.
>> THANK YOU VERY MUCH, JONI,
FOR LAYING DOWN THE CHALLENGES
AND THE VISION SO CLEARLY.
WE APPRECIATE IT.
OKAY.
SO OUR NEXT INTRODUCTORY SPEAKER
IS DR. ROBERT JORDAN, WHO IS A
SENIOR PROGRAM OFFICER FROM THE
BILL AND MELINDA GATES
FOUNDATION.
WELCOME, ROB.
THE FLOOR IS YOURS.
>> GREAT.
THANKS, SARINE.
COULD YOU ACTIVATE MY CAMERA?
IT SAYS THE HOST HAS STOPPED IT.
GREAT.
THANK YOU VERY MUCH.
>> THANK YOU.
>> THANKS, IS SARINE, FOR THAT
INTRODUCTION, AND JONI FOR THE
REALLY NICE INTRODUCTORY
COMMENTS.
I THOUGHT TODAY AS AN
INTRODUCTION, I WANTED TO TALK A
LITTLE BIT ABOUT THE BILL AND
MELINDA GATES FOUNDATION, WHO WE
ARE, WHAT WE DO, AND HOW WE'RE
THINKING ABOUT CULTURES IN THE
DRUG DEVELOPMENT AND DISCOVERY
PROCESS.
I PUT TOGETHER A FEW SLIDE THAT
I'D LIKE TO SHARE.
TO HELP DESCRIBE SOME OF THE
WORK THAT WE'RE DOING IN THE
SPACE.
SO THE DRIVING PHILOSOPHY AT THE
FOUNDATION IS THAT ALL LIVES
HAVE EQUAL VALUE, AND THAT WE
WANT TO DEVELOP PROGRAMS THAT
WILL ENABLE EVERYONE IN THE
WORLD TO LIVE HEALTHY AND
PRODUCTIVE LIVES.
SO WE HAVE REALLY FOUR MAJOR
MISSIONS WE'RE WORKING ON AT THE
FOUNDATION.
FOCUSED ON CHILDREN AND YOUNG
PEOPLE, TO HELP THEM SURVIVE AND
THRIVE, EMPOWER THE POOREST,
ESPECIALLY WOMEN AND GIRLS, TO
HELP TRANSFORM THEIR LIVES.
WE'RE WORKING ON DEVELOPING
THERAPEUTICS, VACCINES,
BIOLOGICS TO TREAT INFECTIOUS
DISEASES, ESPECIALLY THOSE THAT
AFFECT THE POOREST PEOPLE IN THE
WORLD, AS WELL AS INSPIRE PEOPLE
AT THE LOCAL LEVEL TO TAKE
ACTION TO CHANGE, TO CHANGE
LIVES, AND HOW WE DO THIS IS
THROUGH GRANTEES AND PARTNERS,
WHICH ARE THE CENTER OF OUR
WORK.
WE LIKE TO WORK TOGETHER WITH
OUR GRANTEES AND PARTNERS TO
TAKE RISKS, TO PUSH BOUNDARIES,
TO DEVELOP NEW SCIENCE AND ALSO
TO HARNESS THE POWER OF SCIENCE
AND TECHNOLOGY TO DEVELOP
THERAPEUTIC AND OTHER PROGRAMS.
SO THIS WORK DOESN'T HAPPEN IN A
VACUUM OBVIOUSLY, WE LIKE TO
WORK WITH OTHER GOVERNMENTS,
PRIVATE ORGANIZATIONS,
COMMUNITIES, NON-PROFITS AND
INDIVIDUALS TO HELP PROGRESS OUR
PROGRAMS.
ONE OF THE THINGS THAT WE ARE
FOCUSED ON AT THE FOUNDATION IS
GLOBAL HEALTH EQUITY, AND WHAT
WE'VE SEEN IN THE PAST AND MOST
RECENTLY, FOR EXAMPLE, WITH
COVID VACCINES IS WHILE WE ARE
ABLE TO DEVELOP PHENOMENAL
THERAPEUTICS, THEY DON'T ALWAYS
GET TO PEOPLE WHO NEED IT THE
MOST OR GET TO PEOPLE AROUND THE
WORLD IN AN EQUITABLE FASHION.
FOR EXAMPLE, THE MRNA VACCINES
THAT WERE DEVELOPED WHICH ARE
HIGHLY EFFECTIVE AT REDUCING
DISEASE SEVERITY FROM SARS-COV-2
INFECTION AND PREVENTING
INFECTION WERE PRIMARILY
DISTRIBUTED IN HIGH INCOME
COUNTRIES, BUT SIGNIFICANTLY
LAGGED IN THEIR DISTRIBUTION IN
LOW AND MIDDLE INCOME COUNTRIES.
AND SO IN ALL OUR WORK, WE LOOK
FOR GLOBAL SOLUTIONS, PRODUCTS
THAT ARE SUITABLE FOR USE
EVERYWHERE ACROSS THE GLOBE THAT
CAN BE REGISTERED IN A TIMELY
FASHION SO THEY CAN GET TO THE
PEOPLE WHO NEED THEM MOST, AND
WE WANT TO AVOID PRODUCT
NATIONALISM AND WE WANT
ESSENTIALLY TO HAVE EQUAL ACCESS
TO ALL OF THESE THERAPEUTICS.
SO IN ALL OF OUR GRANTS AND OUR
PARTNERSHIPS, WE HAVE THESE
GLOBAL ACCESS PROVISIONS THAT WE
WANT TO FOLLOW TO MAKE SURE THAT
PRODUCTS DEVELOPED CAN BE USED
ACROSS THE GLOBE.
SO SOME OF THE WORK WE'VE DONE
IN THE COVID-19 SPACE AND SOME
OF THE LESSONS WE'VE LEARNED,
FIRST WE REALIZED THAT WE WERE
UNPREPARED, GIVEN THAT THERE WAS
A LACK OF INVESTMENT IN
DEVELOPMENT OF ANTIVIRALS,
ESPECIALLY THOSE TARGETING MANY
OF THE READILY TRANSMISSIBLE
VIRAL FAMILIES.
WE DIDN'T HAVE ANY STRONG
THERAPEUTICS AVAILABLE WHEN
SARS-COV-2 PANDEMIC STARTED.
WE ALSO FOUND AND WE SPENT A
LONG -- INVESTMENT TO REPURPOSE
COMPOUNDS FOR SARS-COV-2.
WHILE THERE WERE SOME FOR SEVERE
AND LATE DISEASE CONTEXT, THERE
WAS VERY FEW THAT COULD BE
REPURPOSED EFFECTIVELY TO TREAT
MILD TO MODERATE DISEASE OR TO
PREVENT A TRANSMISSION IN
POPULATIONS.
SO WE REALIZE THAT THERE'S A
NEED TO FOCUS UPSTREAM IN THE
COURSE OF DISEASE AND DEVELOP
THERAPEUTICS THAT CAN BE READILY
DEPLOYED IN THE EVENT OF AN
OUTBREAK SITUATION.
SO A LOT OF THE WORK THAT WE'RE
FOCUSED ON MOVING FORWARD IS TO
TRY TO DEVELOP ANTIVIRAL
THERAPEUTICS THAT WOULD BE PHASE
2 READY OR CLINIC-READY IN CASE
OF AN OUTBREAK OF A NEW PANDEMIC
THREAT AGENT.
AND THE VIRUS FAMILIES THAT WE
WERE INTERESTED IN LOOKING AT
ARE CORONAVIRUS WITH A MIX O
DEVELOP VIRAL AND HOST TARGETS
WOULD BE DIRECTLY APPLICABLE TO
VIRUS REPLICATION, OR
STIMULATING HOST INNATE
RESPONSES TO PREVENT INFECTION.
AND WE'RE ALSO EXPLORING SOME
OTHER MODALITIES THAT COULD BE
CONSIDERED PROGRAMMABLE, SUCH AS
SIRNAs OR CRISPR TECHNOLOGIES.
WE'RE ALSO IN OTHER ASPECTS OF
THE FOUNDATION LOOKING AT NEW
VACCINES AND BIOLOGICS AS WELL.
AND SO WHAT WE LIKE TO DO IS TO
SUPPORT EARLY SCIENCE AND TOOLS
AVAILABLE ACROSS THE GLOBE FOR
RESEARCHERS TO ENGAGE IN --
PHARMA, BIOTECH, NON-PROFITS,
AND WE HAVE A COMMITMENT TO DATA
SHARING SO THE WORK WE SUPPORT
SHOULD BE AVAILABLE TO
RESEARCHERS ACROSS THE GLOBE.
AND WE ALSO LIKE TO PARTNER WITH
OTHER GOVERNMENT AGENCIES,
ACADEMIC INSTITUTIONS,
COMMERCIAL AND PHILANTHROPIC
FUNDERS TO SUPPORT THIS WORK,
AND FOR EXAMPLE, WE'VE PARTNERED
WITH NOVANORTIS FOUNDATION TO
PUT TOGETHER AN INITIATIVE TO
SUPPORT SMALL MOLECULE DRUG
DEVELOPMENT FOR PANDEMIC THREAT
PATHOGENS.
SO HOW DO WE VIEW ORGANOIDS IN
THIS PROCESS FOR ANTIVIRAL
DISCOVERY?
WE THINK THAT THE ORGANOIDS
REPRESENT A GOOD MODEL SYSTEM
FOR COMPLEX BIOLOGY, AN
INTERMEDIARY BETWEEN THE WHOLE
ANIMAL, PRIMARY TISSUES AND CELL
CULTURAL.
THEY'RE RELATIVELY EASY TO USE,
OBVIOUSLY TECHNICALLY DIFFICULT
TO PUT TOGETHER BUT MORE EASY TO
USE RELATIVE TO PRIMARY TISSUES
AND ANIMALS.
WE FEEL THAT THERE COULD BE A
BETTER TRANSLATION OF THE DATA
GENERATED IN THESE 3D CULTURES
TO HUMAN STUDIES.
IT ALSO HELPS REMOVE SOME OF THE
ETHICAL ISSUES SURROUNDING USE
OF ANIMALS, AND THERE COULD BE
BETTER TRANSLATION OF THE DATA,
GIVEN THAT THERE'S LARGER SAMPLE
SIZES TO IMPROVE STATISTICS FOR
THERAPEUTIC EVALUATION.
SO THESE ARE SOME OF THE IDEAS
THAT WE'RE THINKING WITH IN ABON
TERMS OF THESE 3D CULTURES AND
HOW THEY CAN BE USED IN THE
DISCOVERY PROCESS.
THIS SLIDE SHOWS THE EVOLUTION
OF SOME OF THE ORGANOID OR 3D
CULTURE SYSTEMS OVER TIME.
ON THE RIGHT-HAND SIDE, THERE
ARE EXAMPLES OF ORGANOIDS USE IN
SARS-COV-2 RESEARCH IN DRUG
SCREENING TO IDENTIFY NOVEL
ANTIVIRALS THAT WEREN'T
DISCOVERED IN 2D CULTURES THAT
WOULD BE -- COULD BE
EFFECTIVE -- POTENTIALLY
EFFECTIVE ANTIVIRALS TO TREAT
SARS-COV-2 INFECTIONS, TO STUDY
NEUROTROPISM, 3D CULTURE MODELS
THAT COULD ALSO INCLUDE IMMUNE
COMPONENTS TO UNDERSTAND THE
IMMUNE RESPONSE TO INFECTION, AS
WELL AS JUST STUDIES TO LOOK AT
RECEPTOR USAGE TO UNDERSTAND
PATHOGENESIS OF INFECTION.
SO THERE ARE A LOT OF
APPLICATIONS FOR 3D CULTURES IN
THE ANTIVIRAL DRUG DISCOVERY AND
DEVELOPMENT SPACE.
THE ORGANOIDS, WE HAVE AN
ORGANOID CONSORTIUM AT THE
FOUNDATION THAT'S ACTUALLY
LOOKING AT THE USE OF 3D
CULTURES IN A NUMBER OF
DIFFERENT THERAPEUTIC AREAS WE
ARE INTERESTED IN, AND WE'RE
FOCUSED ON STANDARDIZING AND
BENCHMARKING THESE MODELS,
MAKING PROTOCOLS SCALABLE AND
REPRODUCIBLE SO THAT THEY CAN BE
USED BY OTHER INVESTIGATORS,
DEVELOPING ORGANOID OR 3D
CULTURE MODELS AND CONTAINMENT
LABORATORIES FOR PATHOGEN WORK,
AS ALSO UNDERSTANDING MICRO
ENENVIRONMENT ENGAGEMENT AND
STANDARDIZATION TO VERIFY AND
VALIDATE SOME OF THESE MODEL
SYSTEMS.
SOME OF THE AREAS IN GLOBAL
HEALTH WE'RE INTERESTED IN ARE
OBVIOUSLY ANTIVIRALS, WE'RE
INTERESTED IN NON-HORMONAL
CONTRACEPTIVES, UNDERSTANDING
THE IMPACT OF MICROBIOME IN
SPECIFIC AREAS IN THE GUT AND
THE VA VAGINAL ORGANOID MODELN
BE USED TO CULTIVATE MICROBIOTA,
VACCINE PRODUCTION, AS WELL AS
STANDARD TOXICOLOGY FOR DRUG
DISCOVERY AND DEVELOPMENT ACROSS
THESE DIFFERENT THERAPEUTIC
AREAS.
SO WE HAVE A NUMBER OF QUESTIONS
AND I'M SURE WE'RE GOING TO
EXPLORE MANY OF THESE QUESTIONS
IN THE NEXT TWO DAYS WITH SOME
REALLY FANTASTIC TALKS.
WHERE HAVE ORGANOIDS BEEN MOST
SUCCESSFUL IN THE DRUG DISCOVERY
PROCESS AND FOR WHAT PURPOSE FOR
UNDERSTANDING THE COMPLEX
BIOLOGY, IDENTIFYING NEW TARGETS
AND NEW BIOLOGY THRALD BE THAT E
UNAVAILABLE IN 2D CULTURE
SYSTEMS, ARE THESE BETTER
TRANSLATION FROM THE IN VITRO TO
THE HUMAN SYSTEMS.
AND WHAT GAPS REMAIN IN
REALIZING THE IMPACT OF THESE 3D
CULTURE SYSTEMS AND HOW CAN WE
STANDARDIZE THESE SYSTEMS FOR
DRUG DISCOVERY AND DEVELOPMENT?
AND WHAT WILL IT TAKE TO EXPAND
ACCESS TO AND UTILIZATION OF
THESE MODELS IN DRUG DISCOVERY,
WHERE WOULD THE HIGHEST VALUE BE
IN TERMS OF FUTURE FOR INCREASED
UTILIZATION?
SO THESE ARE A NUMBER OF
QUESTIONS I'M SURE WE'RE GOING
TO BE TALKING ABOUT MANY OF
THESE IN THE TALKS TO FOLLOW.
SO WITH THAT, I'LL TURN IT BACK
TO SARINE.
>> THANK YOU SO MUCH, ROBERT,
FOR THAT WONDERFUL COMMENTS.
OKAY.
SO THE NEXT SPEAKER FOR
INTRODUCTORY COMMENTS IS
DR. EMILYER EMILY ERBELDINGE
NATIONAL INSTITUTES OF ALLERGY
AND INFECTIOUS DISEASES PART OF
THE NATIONAL INSTITUTES OF
HEALTH.
EMILY, THE FLOOR IS YOURS.
>> THANK YOU.
I'M JUST SHARING MY SLIDES AND
I -- HERE WE GO.
SO THANK YOU FOR INVITING ME TO
PRESENT.
I WAS ASKED TO TALK A LITTLE BIT
ABOUT -- TO REFLECT A LITTLE BIT
BACK ON THE LESSONS LEARNED AT
NIAID FROM THE PANDEMIC THAT
WE'RE STILL EXPERIENCING AND A
PATH FORWARD.
IN PARTICULAR, WITH A FOCUS ON
OUR PROTOTYPE PATHOGEN PLAN FOR
ADDRESSING FUTURE PANDEMICS,
READINESS FOR FUTURE PANDEMICS,
WHICH MIGHT OFFER PERSPECTIVE
FOR YOUR DELIBERATIONS OVER THE
NEXT COUPLE DAYS.
SO AT NIAID AND MORE BROADLY IT
WAS RECOGNIZED THAT THERE WERE
NUMEROUS CORONAVIRUS OUTBREAKS
OCCURRING ABOUT EVERY 10 YEARS,
WITH SARS AND THEN MERS, AND WE
NEEDED TO BE PREPARED FOR A
BIGGER CORONAVIRUS WHICH
ACTUALLY HAPPENED IN LATE 2019
AND CONTINUES TO THE PRESENT.
SO IT WASN'T MAGIC, THE RAPID
DEVELOPMENT OF VACCINES WASN'T
MAGICAL.
THERE WAS A LOT OF SCIENTIFIC
EFFORT AND INVESTMENT THAT WENT
IN A MULTIDISCIPLINARY
INVESTMENTS IN STRUCTURAL
BIOLOGY FOR CORONAVIRUS BECAUSE
PEOPLE RECOGNIZED THAT THIS TYPE
OF VIRUS HAD PANDEMIC POTENTIAL
AND THE THREAT WAS THERE FOR AN
UNPRECEDENTED PANDEMIC.
SO I'M JUST GOING TO TELL YOU A
LITTLE BIT ABOUT THIS STORY
WHICH HAPPENED WITHIN NIAID AT
THE VRC ALONG WITH EXTRAMURAL
INVESTIGATORS.
BACK MORE THAN 10 YEARS AGO, IT
WAS RECOGNIZED BY BARNEY GRAHAM,
WHO'S NOW RETIRED FROM THE VRC,
THAT STRUCTURAL BIOLOGY DEFINING
PARTICULARLY WITH RSV ENTRY
PROTEINS, IF YOU COULD STABILIZE
AN ENTRY PROTEIN IN ITS
PRE-FUSION STRUCTURE USING
VARIOUS TOOLS, YOU COULD IMPROVE
ON ITS IMMUNOGENICITY AND THAT
COULD PROVIDE A BASIS FOR A
BETTER VACCINE FNLT SO THERE WAS
SO THIS WAS ALSO APPLIED TO THE
CORONAVIRUS SPIKE PROTEIN OF
MERS, AND THIS WAS BACK 10 YEARS
AGO, SO INTRODUCING A FEW
MUTATIONS AT A CERTAIN PART OF
THE SPIKE PROTEIN, YOU COULD
STABILIZE IT IN ITS PRE-FUSION
COMPLEX AND RATHER THAN
INFUSING -- USING AS AN ANTIGEN
VARIOUS CONFIRMATIONS OF THE
PROTEIN, YOU COULD GENERATE A
VERY PRECISE, BETTER ANTIBODIES,
MORE PRECISELY TARGETED TO THE
SPIKE PROTEIN THAT WOULD BE MORE
IMMUNOGENIC THAN IF IT WASN'T
HELD IN ITS PREFUSION
CONFORMATION.
SO VACCINES WERE IN THE EARLY
STAGES OF DEVELOPMENT.
IT WAS ALSO TESTED ON A SEASONAL
CORONAVIRUS WHICH ALSO HAD A
SPIKE PROTEIN, THE
OC43 CORONAVIRUS AND IT WAS
HYPOTHESIZED THAT THAT COULD BE
A GENERALIZABLE SOLUTION TO
DEVELOP A VACCINE FOR ANY
CORONAVIRUS.
SO AGAIN, ALL THE CORONAVIRUS
HAVE THESE SPIKE PROTEINS,
PUTTING SPECIFIC MUTATIONS AND
SUBSTITUTING PROLINES IN A FEW
SITES LED TO A STABLE PRE-FUSION
FORM THAT PROVIDED THE BASIS FOR
FIVE OF THE SIX VACCINES THAT
WERE SUPPORTED IN DEVELOPMENT
FOR THE SARS-COV-2 PANDEMIC.
SO THIS SHORTENED TREMENDOUSLY
THE STORY LINE WE HEAR IN THE
PRESS IS SORT OF THAT IT WAS THE
MIRACLE OF MRNA VACCINE PLATFORM
THAT LED US TO BE SO RAPID IN
OUR VACCINE DEVELOPMENT.
BUT REALLY IT WAS YEARS AND
YEARS OF THESE MULTIDISCIPLINARY
TEAMS DEFINING THE STRUCTURE,
TESTING DIFFERENT SUBSTITUTIONS
TO PUT THE PROTEIN, THE ENTRY
PROTEIN OF SARS, WELL, SARS
1 ORIGINALLY BUT THEN OTHER
CORONAVIRUS, TO GENERATE -- TO
USE THAT STABILIZED PRE-FUSION
PROTEIN TO GENERATE AN IMMUNE
RESPONSE.
AND THAT ALLOWED -- THOSE YEARS
OF BASIC SCIENCE WORK AND
TRANSLATIONAL -- EARLY
TRANSLATIONAL WORK LED TO THE
CORONAVIRUS VACCINE BEING
DEVELOPED IN SUCH RAPID TIME,
ALONG WITH THE ADVENT OF THE
MRNA PLATFORM.
SO THIS IS SORT OF THIS IDEA AND
THE SUCCESS OF THIS, YOU KNOW,
EARLY BASIC SCIENCE DISCOVERY,
EARLY PRODUCT DEVELOPMENT AS DES
SORT OF BEEN DEVELOPED INTO, I
GUESS YOU'D SAY, AN APPROACH TO
PANDEMIC PREPAREDNESS THAT NIAID
HAS NOW ENDORSED AS A STRATEGIC
PLAN FOR PREPARING -- FOR
RESEARCH THAT WOULD PREPARE US
FOR FUTURE PANDEMICS.
THE IDEA GOES SOMETHING LIKE
THIS, IF YOU THINK ABOUT VIRUSES
THAT ARE OUT THERE THAT COULD
SPILL FROM ANIMAL RESERVOIRS
INTO THANK HUMANS, IT SEEMS E
NUMBER IS INFINITE AND THE
THREAT IS INFINITE BUT IF YOU
BREAK THAT DOWN INTO 20 VIRAL
FAMILIES THAT ARE RESPONSIBLE
FOR HUMAN DISEASE AND IF YOU
BREAK THAT DOWN FURTHER TO THE
RNA VIRUSES THAT ARE LIKELY TO
CAUSE SPILLOVER EVENTS BECAUSE
OF THEIR HIGH MUTATION RATE,
REALLY THERE'S LESS THAN 10
VIRAL FAMILIES THAT PROBABLY
HAVE PANDEMIC POTENTIAL.
IF YOU COULD REPLICATE THE
STEPS, THE EARLY SCIENCE STEPS
IN SCIENCE THAT LED TO THE RAPID
DEVELOPMENT OF THE MRNA VACCINES
AND THE OTHER GENE-ENCODED
VACCINES, IF YOU REPLICATE THOSE
STEPS WITH DEFINING AN ANTIGENIC
TARGET TESTING ANIMALS AND ALSO
TESTING MONOCLONAL ANTIBODIES
TESTING THAT SPECIFIC STRUCK
TOOR FOR VIRAL ENTRY, YOU MIGHT
BE WAY FARTHER ALONG SHOULD
ANOTHER VIRUS IN THAT FAMILY
BECOME -- CAUSE AN OUTBREAK THAT
MIGHT LEAD TO AN EPIDEMIC OR
EVEN A PANDEMIC.
SO THIS, WE'RE CALLING THE
PROTOTYPE PATHOGEN APPROACH TO
ACCELERATE MEDICAL
COUNTERMEASURES, VACCINES AND
ALSO MONOCLONAL ANTIBODIES, SO
CHOOSING THESE PROTOTYPE
PATHOGENS WITHIN THE VIRAL
FAMILIES HAS BECOME A FOCUS OF
OUR THINKING OVER THE PAST
SEVERAL MONTHS.
SO OUTLINED ON THE SLIDE
REPRESENTED ARE THE VIRAL
FAMILIES OF CONCERN.
THEY'RE ALL RNA VIRUSES BECAUSE
WE KNOW THESE ARE THE VIRUSES
THAT HAVE HIGH MUTATIONAL RATES
AND, THEREFORE, WITH A LOT OF
MISTAKES, MIGHT LEAD TO
SOMETHING THAT BECOMES MORE
TRANSMISSIBLE SOMETIMES OR MORE
LIKELY TO BE PATHOGENIC IN
HUMANS AND WHEN HUMANS COME INTO
PROXIMITY WITH THEIR ZOONOTIC
RESERVOIR, MORE LIKELY TO CAUSE
DISEASE, POTENTIALLY CAUSE AN
OUTBREAK AND MAYBE EVEN CAUSE A
PANDEMIC.
SO CORONAVIRUS AND ORTHO
MYXOVIRADE HAVE BEEN OF CORNER
FOR A WHILE BUT THERE ARE OTHER
VIRUSES THAT ARE LESS UNDERSTOOD
THAT HAVE FEWER VIRGINIA BEACH
INVESTMENTS BUT PROBABLY STILL
HAVE EPIDEMIC AND MAYBE EVEN
PANDEMIC POTENTIAL.
SO OUR RESEARCH STRATEGY OVER
UPCOMING YEARS, BOTH INTRAMURAL
AND EXTRAMURAL AT NIAID FOR
PANDEMIC PREPAREDNESS, IS TO
FOCUS ON THESE OTHER VIRAL
FAMILIES.
SO WE HAVE PROPOSED, WE HAVEN'T
EVEN GOTTEN FUNDING TO FUND
THESE CENTERS THAT WOULD DEVELOP
VACCINES AND MONOCLONAL
ANTIBODIES FOR PANDEMIC
PREPAREDNESS, WE HAVE PROPOSED A
FUND FOR MULTIDISCIPLINARY TEAMS
EARLY WORKING ON CORONAVIRUS
THAT LED TO RAPID DEVELOPMENT OF
VACCINES AND THEN ALSO
MONOCLONAL ANTIBODIES, AND THEN
WE HOPE THAT THIS APPROACH
GENERALLY CAN ALSO TRANSLATE
INTO PROGRESS IN IDENTIFYING
TARGETS FOR ANTIVIRAL -- OTHER
TYPES OF ANTIVIRALS SUCH AS
SMALL MOLECULE THERAPEUTICS.
SO WE PROPOSE TO DEVELOP CENTERS
THAT FOCUS ON BASIC VEERLOLOGY,
STRUCTURAL BIOLOGY, ANTIGEN
MAPPING, ET CETERA, ANIMAL
MODELS AND REPLACEMENT MODELS
FOR -- MODELS THAT WOULD REPLACE
ANIMALS, AS WELL AS IND-ENABLING
RESEARCH TO FOCUS ON PROTOTYPE
PATHOGENS FROM EACH OF THESE
FAMILIES AND IDEALLY THEY WOULD
FOCUS ON MORE THAN ONE.
SO FOR THE FUTURE, WE BELIEVE
THAT THAT WOULD PROVIDE A PATH
TOWARDS BEING PREPARED FOR
POTENTIALLY ANY ZOO NO ZOON-
THAT MIGHT LEAD TO A PANDEMIC.
SO I WAS ASKED TO SUMMARIZE SOME
OF THE LESSONS WE'VE LEARNED TO
DATE FROM THE COVID-19 PANDEMIC.
WE'RE STILL LEARNING, OF COURSE,
REALLY THE FIRE IS STILL
SMOLDERING, THIS ISN'T DONE SO
IT FEELS A LITTLE BIT ODD TO
REFLECT AS THOUGH IT WERE DONE,
BUT SOME LESSONS LEARNED, YEARS
OF VIROLOGY AND IP MU NOLG AND
STRUCTURAL BIOLOGY PREPARED THE
WAY FOR RAPID VACCINE
DEVELOPMENT AND IF IT WAS A
DIFFERENT VIRAL FAMILY, WE COULD
NOT HAVE RESPONDED AS RAPIDLY OR
EFFECTIVE, DUE TO THE KNOWLEDGE
WE HAD FOR CORONAVIRUS WHEN THE
SARS-COV-2 PANDEMIC STARTED.
THE RECENT EMERGENCE OF MRNA AS
AN IMMUNOGENIC PLATFORM WAS KEY
TO RAPIDLY MANUFACTURING A
VACCINE AND KEY TO THE CLINICAL
RESPONSE, BUT IT WASN'T A
MIRACLE.
WE ONLY KNEW WHAT TARGET TO
ENCODE IN THE MRNA BECAUSE OF
YEARS OF BASIC SCIENCE AND
TRANSLATIONAL SCIENCE AND WORK
THAT WAS DONE TO BUILD UP TO
THAT POINT.
SO WE PROPOSE THAT THE WORLD
NEEDS A MORE SYSTEMATIC APPROACH
TO PANDEMIC PREPAREDNESS, AND
THIS MIGHT INVOLVE FOCUSING ON
PROTOTYPE PATHOGENS WITHIN EACH
OF THESE VIRAL FAMILIES WITH
PANDEMIC POTENTIAL.
SO IN ADDITION TO FOCUSING ON
VACCINE DEVELOPMENT, MONOCLONAL
ANTIBODY DEVELOPMENT WHICH HAVE
SOME SCIENTIFIC SIMILARITIES
EARLY IN THE DEVELOPMENT STAGE,
WE ALSO ARE TRYING TO FOCUS ON
ANTIVIRAL DEVELOPMENT IN THE
SAME WAY, SO RECENTLY WE
ANNOUNCED FUNDING FOR ANTIVIRAL
DISCOVERY CENTERS.
THESE ARE LARGE COOPERATIVE
AGREEMENTS FUNDED THROUGH THE
AMERICAN PANDEMIC PREPAREDNESS
PROGRAM.
THESE ARE LARGE
MULTIDISCIPLINARY TEAMS THAT
WILL FOCUS ON THE SAME THINGS
THAT PROVIDED A FOUNDATION FOR
RAPID DEVELOPMENT OF THE
SARS-COV-2 VACCINE, STRUCTURAL
BIOLOGISTS, ME , MEDICINAL CHE,
IN VITRO TESTING, ANIMAL MODEL
DEVELOPMENT.
THEY WILL FOCUS ON SARS-COV-2
AND OTHER CORONAVIRUS BUT ALL OF
THEM WILL HAVE ONE ADDITIONAL
VIRAL FAMILY WITHIN THESE GROUPS
THAT MIGHT HAVE PANDEMIC
POTENTIAL, BECAUSE WE RECOGNIZE
THAT SOME THERAPEUTICS MIGHT
HAVE EFFECTIVENESS ACROSS VIRAL
FAMILIES.
SO THAT'S OUR PLAN.
AND I'M HAPPY TO TAKE QUESTIONS
IF HE THERE'S TIME.
THANK YOU.
>> THANK YOU VERY MUCH, EMILY.
I APPRECIATE THOSE WONDERFUL
COMMENTS THERE.
THERE IS ACTUALLY ONE QUESTION
FOR YOU SO I'LL JUST READ IT FOR
YOU.
SO THE QUESTION SAYS, IT HAS
BEEN SHOWN THAT THE IMMUNE
REPERTOIRE HAS AN IMPACT ON THE
EFFICACY OF MRNA VACCINES.
IS THE SAME TRUE FOR OTHER TYPES
OF VACCINES?
>> PROBABLY.
THE IMMUNE REPERTOIRE, I DON'T
KNOW SPECIFICALLY WHAT ASPECT OF
THE IMMUNE RESPONSE THE
QUESTIONER IS FOCUSED ON, BUT I
THINK WE KNOW THAT THE
NON-REPLICATING VIRAL VECTOR
VACCINES PROBABLY STIMULATE A
BETTER T-CELL RESPONSE AND
LESS -- AND PROTECT EVEN THOUGH
THEY ARE STIMULATING --
GENERATING A LOWER AMOUNT OF
HUMERAL ANTIBODIES, SO
NEUTRALIZING ANTIBODIES, SO IT
MUST BE, SO YES, OTHER THAN MRNA
VACCINES, THERE IS A DIFFERENCE
IN THE IMMUNE REP REPERTOIRE S
STIMULATED.
>> THANK YOU VERY MUCH, EMILY.
APPRECIATE YOUR TIME.
OKAY.
SO WITH THE NEXT FEW MINUTES,
I'M GOING TO GO OVER THE MISSION
OF THE ASSAY GUIDANCE PROGRAM
AND I'LL TALK MORE ABOUT THE
WORKSHOP OBJECTIVES.
SO LET ME SHARE MY SLIDES.
SO AS I SAID EARLIER, I'M THE
EDITOR-IN-CHIEF OF THE ASSAY
GUIDANCE MANUAL AND THE LEAD OF
THE TRANSLATIONAL SCIENCE
RESOURCES PROGRAM HERE AT NCATS.
SO BASICALLY WHY WE ARE DOING
WHAT WE DO IN OUR PROGRAM IS
BECAUSE THERE IS A CRISIS OF EAR
IRREPRODUCIBILITY IN BASIC AND
PRE-CLINICAL RESEARCH THAT MANY
OF YOU ARE I'M SURE AWARE.
A PUBLICATION IN 2015 SHOWED
THAT THE CUMULATIVE PREVALENCE
OF EAR REPRODUCIBLE PRE-CLINICAL
RESEARCH EXCEEDS 50%, AND THIS
ACTUALLY RESULTS IN
APPROXIMATELY $28 BILLION U.S.
SPENT ON PRE-CLINICAL RESEARCH
THAT IS NOT REPRODUCIBLE.
AND THE CATEGORIES OF
PRE-CLINICAL EAR REPRO
QUEUESIBILITY INCLUDE BIOLOGICAL
REAGENTS AND REFERENCE
MATERIALS, STUDY DESIGN, DATA
ANAL
ANALYSIS AND REPORTING AND DATA
AND LABORATORY PROTOCOLS.
THE ASSAY GUIDANCE MANUAL
PROGRAM THAT I LEAD IS A DISEASE
AGNOSTIC PROGRAM THAT ADDRESSES
THIS CRISIS OF IRREPRODUCIBILITY
IN BASIC AND PRE-CLINICAL
RESEARCH AND WE DO SO BY SETTING
STANDARDS FOR RI CORE RIGORN
TRANSLATIONAL SCIENCE.
WHAT WE ACTUALLY DO IS WE
PROVIDE MULTIPLE RESOURCES WITH
THE OVERALL GOAL OF HELPING
SCIENTISTS LIKE YOU DESIGN AND
IMPLEMENT ROBUST ASSAYS IN EARLY
TRANSLATION RESEARCH.
THE MAIN COMPONENT OF OUR
PROGRAM IS THE ASSAY GUIDANCE
MANUAL, WHICH IS A FREE AND
PUBLICLY AVAILABLE RESOURCE AND
IS ACCESSIBLE AS AN EBOOK ON THE
NATIONAL LIBRARY OF MEDICINE
NCBI BOOKSHELF.
THIS EBOOK IS DYNAMIC WITH
CONTINUED RENEWAL AND EXPANSION
OF CONTENT, AND IT IS WIDELY
ACCESSED AS WE GOT MORE THAN
40,000 VIEWS PER MONTH IN 2022.
AND IT IS ACTUALLY ACCESSED BY
SCIENTIST FROM PHARMA, BIOTECH,
GOVERNMENT AND ACADEMIA ALIKE.
WE ALSO RUN WORKSHOPS AS PART OF
OUR PROGRAM, AND THIS IS ONE
TOPIC-FOCUSED WORKSOP THAT WE WE
ARE RUNNING.
SO WHAT I'D LIKE TO DO NEXT IS
GO OVER HIGH LEVEL GOALS OR
OBJECTIVE FS THIS WORKSHOP.
SO THE HIGH LEVEL GOALS OF THIS
WORKSHOP IS TO HELP SCIENTISTS
ESTABLISH ROBUST REPRODUCIBLE
SCALABLE CONSISTENT ADVANCED
TISSUE MODELS TO STUDY PANDEMIC
THREAT VIRUSES.
WE'D LIKE TO PROVIDE YOU ALL
WITH PRACTICAL APPROACHES AND
BEST PRACTICES FOR DEVELOPING
STANDARDIZED ASSAYS WITH THE
HOPE OF HELPING YOU TO
SUCCESSFULLY DEVELOP
THERAPEUTICS FOR FUTURE PANDEMIC
THREATS, AND WE WOULD REALLY
LIKE TO GENERATE A ROAD MAP TO
HELP SCIENTISTS DEVELOP ROBUST
AND REPRODUCIBLE 3D TISSUE
MODELS TO STUDY VIRUSES.
THE SPECIFIC ABTTIVES OF THIS
WORKSHOP INCLUDES INTRODUCE
PARTICIPANTS TO THE AVAILABLE 3D
TISSUE MODELS AND DISCUSS
CHALLENGES IN BUILDING THESE
MODELS AS WELL AS THEIR UTILITY
AND LIMITATIONS, AND MOST OF
THIS FIRST OBJECTIVE WILL BE
COVERED IN TODAY'S SESSION, ONE
IN THE AFTERNOON.
WE'D ALSO LIKE TO PROVIDE CASE
STUDIES ON HOW THESE AVAILABLE
3D TISSUE MODELS HAVE BEEN
UTILIZED IN ANTIVIRAL DRUG
DISCOVERY AND DEVELOPMENT AS
WELL AS TOOLS FOR UNDERSTANDING
AND MODELING INFECTIOUS
DISEASES, INCLUDING STUDYING
VIRAL INFECTION AND
PATHOGENESIS, AND THIS OBJECTIV
OBJECTIVE -- MOST OF THESE WILL
BE COVERED IN SESSION TWO
TOMORROW, AND WE'D ALSO LIKE TO
PROVIDE GUIDELINES AND
CONSIDERATIONS FOR DEVELOPING
ROBUST AND REPRODUCIBLE 3D
TISSUE MODELS THAT CAN BE US
OOOD FOR TESTING THERAPEUTICS.
WE'D LIKE TO DISCUSS CHALLENGES
IN AFFORDABILITY, ACCESSIBILITY,
TRANSFERABILITY AND
REPRODUCIBILITY OF THESE 3D
TISSUE MODELS, AND ALSO DISCUSS
THE ADVANTAGES VERSUS THE
DISADVANTAGES OF THESE 3D MODELS
WILL BE COVERED IN SESSION ONE,
TWO AS WELL AS SESSION THREE.
OKAY.
I ALWAYS ASK FOLKS TO CONNECT
WITH US TO STAY TUNED ABOUT ANY
UPCOMING WORKSHOPS AND SYMPOSIA
WE HAVE IN OUR PROGRAM, AS WE
HAVE ONE COMING UP IN OCTOBER,
THIS IS AN IN-PERSON WORKSHOP ON
ASSAY GUIDANCE FOR
HIGH-THROUGHPUT SCREENING AND
LEAD DISCOVERY.
IT WILL BE RUN IN NIH MAIN
CAMPUS, AND THERE IS MULTIPLE
WAYS CAN YOU CONNECT WITH US.
WE ARE ON TWITTER AND OUR
TWITTER HANDLE IS@ASSAYGUIDANCE.
WE HAVE A LINKEDIN GROUP THAT
YOU CAN JOIN HERE, AS WELL AS
YOU CAN SEE ALL OUR EVENTS AT
NCATS EVENTS WEBSITE, AND NCATS
AGM WEBSITE AS WELL.
SO BACK TO THE MAIN EVENT,
TODAY'S AGENDA HERE THAT I'M
SHARING.
SO AFTER THIS, WE'LL HAVE THE
KEYNOTE SPEECH BY DR. SIMON
FUNNELL.
WE'RE REALLY EXCITED TO HEAR HIM
SPEAK TODAY.
AND THEN WE'LL HAVE A LUNCH
AFTER HIS KEYNOTE, AND THEN THIS
AFTERNOON WE'LL HAVE THE FIRST
SESSION, WHICH IS ON 3D TISSUE
MODELS, UTILITY AND LIMITATIONS,
AND WE WILL CONCLUDE THE DAY
TODAY AROUND 4:15.
I DO WANT TO SAY THAT, PLEASE
FEEL FREE TO ASK ANY QUESTIONS
YOU HAVE THROUGHOUT THE TALKS
USING THE Q & A CHAT BOX
FUNCTION, AND THE SPEAKERS WILL
GET TO YOUR QUESTIONS AT THE END
OF THEIR -- FOR EXAMPLE, THE
KEYNOTE SPEAKER WILL GET TO YOUR
QUESTIONS AT THE END OF HIS
SPEECH, AND THEN FOR THE
SESSIONS, WE WILL HAVE A Q & A
DISCUSSION SESSION AFTER ALL THE
TALKS HAVE COMPLETED, BUT FEEL
FREE TO ASK YOUR QUESTIONS AND
IF YOU WOULD LIKE TO DIRECT YOUR
QUESTIONS TO A PARTICULAR
SPEAKER, MAKE SURE YOU ADD THE
NAME OF THE SPEAKER WHEN YOU
TYPE IN THE QUESTION IN THE Q &
A BOX.
OKAY.
I'M EXTREMELY EXCITED TO
INTRODUCE OUR KEYNOTE SPEAKER,
DR. SIMON FUNNELL, A SCIENTIFIC
LEADER BASED AT UK HEALTH
SECURITY AGENCY.
HE ALSO HAS A POSITION AT THE
QUANTUM INSTITUTE IN NORWICH,
ALSO A FACULTY MEMBER AT THE
UNIVERSITY OF TEXAS MEDICAL
BRANCH AND HAS BEEN SECONDED
PART-TIME TO THE R & D
BLUEPRINTING AT THE WORLD HEALTH
ORGANIZATION IN GENEVA SINCE
FEBRUARY 2020.
DR. FUNNELL HAS 36 YEARS OF
WORKING EXPERIENCE AND HAS
STUDIED AND PUBLISHED PAPERS ON
A WIDE RANGE OF SELECT AGENTS.
HE'S EXPERIENCED WITH WORKING
WITH OTHER DANGEROUS PATHOGENS,
ASSISTED THE U.K. HEALTH
SECURITY AGENCY TO PIVOT THEIR
RESOURCES SO THE SARS-COV-2
RESPONSE AND ASSURED THAT THE
U.K. HEALTH SECURITY AGENCY WAS
AND CONTINUES TO BE REACTIVE TO
THE DYNAMIC SITUATION PRESENTED
TO US ALL.
YOU AM SUPER EXCITED TO HAVE YOU
WITH US HERE TODAY, SIMON, AND
WE ARE REALLY EXCITED TO HEAR
YOUR TALK.
LIKE I SAID EARLIER, PLEASE FEEL
TBREE TO ASK ANY QUESTIONS YOU
HAVE FOR DR. FUNNELL THROUGH THE
Q & A CHAT BOX AND HE WILL GET
TO THESE QUESTIONS AT THE END OF
THE TALK.
THANK YOU SO MUCH, SIMON, AGAIN,
AND THE FLOOR IS YOURS.
>> THANK YOU VERY MUCH, SARINE,
FOR THAT INTRODUCTION.
AND THANK YOU ALSO TO ALL THE
ORGANIZERS FOR INVITING ME TO
SPEAK TO YOU TODAY ABOUT
SARS-COV-2, HOW IT'S THE START
OF A NEW SCIENTIFIC RENAISSANCE
IN BIOLOGICAL SCIENCE.
CAN I CHECK THAT THAT'S IN
PRESENTATION MODE?
>> IT IS NOT.
THERE WE GO.
>> GREAT.
THANK YOU VERY MUCH.
SO AS YOU'VE MENTIONED, I HAD
THREE POSITIONS, THANK YOU,
SARINE, FOR INTRODUCING ME IN
THAT WAY.
I HAVE FIVE COMPONENTS OF THE
TALK I'D LIKE TO GIVE TODAY.
FIRST I'D LIKE TO TALK ABOUT WHY
I THINK WE'RE IN THE MIDDLE OF A
21ST CENTURY SCIENTIFIC
RENAISSANCE, THEN I'D LIKE TO
TALK ABOUT ASPECTS OF INFECTION
MORE GENERALLY, AND THEN I'LL
TALK ABOUT MODEL SUITABILITY,
WHAT ARE THE THREE MAJOR COMPLEX
REQUIREMENTS OF AN INFECTIOUS
DISEASE MODEL.
THEN I'LL GIVE SOME EXAMPLES,
BOTH OLD AND NEW, AND THEN TALK
ABOUT SUPPLY AND DEMAND ISSUES.
SO TO BEGIN WITH, WHY DO I SAY
THAT WE'RE IN THE 21ST CENTURY
SCIENTIFIC RENAISSANCE?
I'VE HAD A LOOK AT DICTIONARY
DEFINITIONS OF RENAISSANCE, AND
TRADITIONALLY, IT'S BEEN IN
REFERENCE TO THE 15TH, 16TH
CENTURIES IN TERMS OF GROWTH OF
ACTIVITY IN ARTS AND LITERATURE.
AND I'VE LOOKED AT SOME OF THE
USAGE OF THE WORD, AND I'D LIKE
TO PROPOSE A NEW DEFINITION TO
THE WORD RENAISSANCE.
I BELIEVE THAT WE'RE IN THE
MIDDLE OF A GLOBAL -- DURING THE
GLOBAL SARS-COV-2 OUTBREAK THERE
WAS SUCH ENORMOUS GROWTH IN
SCIENTIFIC RESEARCH,
COLLABORATION AND ACTIVITIES, I
THINK THAT CONSTITUTES A
SCIENTIFIC RENAISSANCE.
AND AN EXAMPLE OF ITS USE COULD
BE, "I AM PARTICIPATING IN A
21ST CENTURY SCIENTIFIC
RENAISSANCE IN WHICH SCIENTISTS,
DEVELOPERS AND FUNDERS ARE
COLLABORATING USING THE POWERS
OF MICRO PHYSIOLOGICAL SYSTEMS."
TO EXPAND ON THIS IDEA THAT WE
ARE IN THE MIDDLE OF A
SCIENTIFIC RENAISSANCE, JUST
LOOK AT WHAT'S HAPPENED IN TERMS
OF GLOBAL SOCIETY.
THERE'S BEEN GLOBAL TRAVEL BANS,
LOCKDOWNS AND AWARENESS IN THE
GLOBAL POPULATION THAT HAS
REALLY FOCUSED ATTENTION ON
BIOLOGICAL SCIENCE.
WE'VE HAD GLOBAL PUBLIC
ENGAGEMENT AND PERSONAL
PROTECTIVE EQUIPMENT AND
LOCKDOWNS.
WIRELESS BANKING HAS TRANSFORMED
THE WAY THAT WE DO BUSINESS, AND
THAT DOESN'T APPEAR TO BE
CHANGING.
REMOTE WORKING HAS BECOME THE
NORM, EVEN THOUGH WE'RE NOW
APPEARING TO GO BACK TO
NORMALITY, THERE'S FAR LESS
COMMUTING THAN THERE EVER WAS.
WE'VE ALSO SEEN AN EPIDEMIC OF
DISINFORMATION, WHICH AT FIRST
SEEMED TO BE EAR EAR RESIST T.
THEY NOW REALIZE THAT THEY HAVE
TO GO TO SCIENTIFIC INFORMATION
TO FORM THEIR JUDGMENTS AND NOT
RELY ON SOCIAL MEDIA.
SO THE IMPORTANCE OF BIOLOGICAL
SCIENCE HAS REALLY BLOSSOMED AND
SO HAS THE IMPORTANCE OF
BIOLOGICAL SCIENTISTS.
POLITICIANS HAVE HAD TO RESPOND
TO PUBLIC CONCERNS ABOUT SARS
COV-2 AND NOW WE SEE POLITICIANS
WITH A THIRST FOR DATA-DRIVEN
DECISIONS AND THAT'S BECOME
INCREASINGLY IMPORTANT FOR
POLITICIANS.
WE'VE SEEN GLOBAL DATA SHARING
ON AN UNPRECEDENTED LEVEL AND
WE'VE SEEN LIVE DATA SHARING
THREE TIMES A WEEK COORDINATED
BY THE WHO AND IN SOME TELECOMS
OVER 1 1/2 THOUSAND PARTICIPANTS
IN CONSORTIUMS WHICH HAVE BEEN
HAPPENING QUITE FREQUENTLY.
THIS REALLY IS UNPRECEDENTED
COLLABORATION.
I THINK WE ALL UNDERSTAND THESE
COMPLEX PROBLEMS WE FACE FROM
DISEASES LIKE SARS-COV-2 REALLY
DO NEED CROSS WORKING NOT JUST
BETWEEN COUNTRIES BUT BETWEEN
THE FACULTIES OF SCIENCE WHICH
WE SEEM TO BE BRINGING TOGETHER
VERY EFFECTIVELY IN THESE TYPES
OF FORUMS.
AS WE'VE JUST HEARD FROM ALL THE
PREVIOUS SPEAKERS, WE'VE SEEN
SOME AMAZING DEVELOPMENTS IN 3D
MODELING TECHNOLOGY AND
ADVANCES.
AND AS A RESULT OF THAT, WE ARE
NOW BEGINNING TO SEE LEGISLATIVE
CHANGES WHICH ARE HAPPENING AS
WE SPEAK, WHICH ARE GOING TO
ENABLE US TO ACCEPT DATA
GENERATED IN THESE TYPES OF
SYSTEMS.
I THINK THAT'S WHY I THINK WE'RE
IN THE MIDDLE OF A SCIENTIFIC
RENAISSANCE.
SO MOVING ON, I'D LIKE TO TALK
ABOUT ASPECTS OF INFECTION IN
RESPECT TO MY EXPERIENCE IN THE
LAST 36 YEARS.
I THINK BROADLY, YOU CAN TALK
ABOUT A PATHOGEN AND BREAK DOWN
THE STEPS THAT A PATHOGEN
INTERACTS WITH A HOST WITH,
WHICH LEADS TO DISEASE.
AND THE VERY FIRST OF THOSE IS
CONTACT.
AND I KNOW IN THE INTRODUCTORY
SPEAKERS, WE'VE BEEN HEARING
ABOUT HOW THAT THAT WAS THE T
FOR MANY YEARS AND INDEED THAT'S
AN EFFECTIVE APPROACH.
ONE AN ORGANISM HAS HAD COULD BE
TACT WITH A CONTACT
WITH A HOST IT NORMALLY FORMS A
MORE SECURE ATTACHMENT.
FOLLOWING ATTACHMENT, AN EVEN
MORE SECURE ADHESION.
WE OFTEN SEE PATHOGENS MIMICKING
THE HOST AND IN THE CASE OF
VIRUSES, WHICH I'VE PUT IN
ITALICS, WE THEN SEE INNOVATION
OF THE HOST CELL, TRANSCRIPTION
OF THE PATHOGEN GENETIC
MATERIAL, PIRACY OF THE HOST,
THEN INTERFERENCE WITH THE HOST
MACHINERY AND IMMUNE RESPONSE.
AND IF IT EFFECTIVE, WE THEN SEE
PATHOGEN ASSEMBLY WITHIN THE
HOST CELL, WHICH LEADS TO
REPRODUCTION OF THE PATHOGEN,
AND THEN WHICH COULD LEAD TO
DISSEMINATION OF THE PATHOGEN TO
OTHER HOSTS, OR CONTINUATION OF
DISEASE IN THE ORIGINAL HOST.
AND I THINK BREAKING IT DOWN
INTO THOSE STEPS HELPS US TO
THINK ABOUT WAYS IN WHICH A HOST
RESPONDS TO INFECTION.
SO HERE I'VE TRIED TO BREAK DOWN
SOME OF THE KEY HOST RESPONSES
TO THOSE PROCESSES THAT A
PATHOGEN REPRESENTS.
THE FIRST OF THOSE IS AVOIDANCE.
WE ALL KNOW HOW TO AVOID
SARS-COV-2, WE ISOLATE AND PUT
IN PERSONAL PROTECTIVE
EQUIPMENT, BUT THERE'S ALSO --
WE HAVE AN INNATE INTEGMEN,
WHICH AVOID THE PATHOGEN MAKING
ATTACHMENT TO OUR HOST CELLS.
AND PART OF THAT, I WOULD
INCLUDE THE MUCOSAL MUCUS FLOW
WHICH IS INCREDIBLY COMPLEX.
IF WE THINK ABOUT THE HUMAN
AIRWAY, WE HAVE WITHIN OUR LUNGS
WHAT WE CALL A MUCOCILIARY
ESCALATOR.
THE WALLS OF OUR TRACHEA AND
BRONCHI ARE LINED WITH CILIATED
EPITHELIUM AT TWO HUP BEATS PER
SECOND.
TOGETHER THERE'S AN ORCHESTRATED
MOVEMENT OF MUCUS UP TO OUR
THROAT, WHERE WE CAN EITHER
EXPECTORATE, SWALLOW OR
ELIMINATE THE PATHOGEN FROM OUR
RESPIRATORY ALVEOLI.
WHAT'S INTERESTING ABOUT THESE
CELLS, IT HAS MASSIVE METABOLIC
IMPACT ON THOSE CELLS AND IT'S
INTERESTING THAT THOSE ARE THE
CELLS THAT SARS-COV-2 TARGETS.
WHEN WHEN I SAY INTEGUMEN, THE
SKIN HAS EPITHELIAL CELLS,
ENDOTHELIAL CELLS, DENDRITIC
CELLS, HAIRS, THE FOLLICLES OF
THE HAIRS, AND WE HAVE SENSORY
CONNECTIONS TO THE HAIR,
MUSCULAR CONNECTIONS TO THE
HAIR, SEBACEOUS GLANDS, SWEAT
GLANDS, VENOUS AND ARTERIAL
BLOOD SUPPLY, ADIPOSE TISSUE,
AND ON TOP OF ALL OF THAT
COMPLEXITY, THE IMMUNE SYSTEM
AND IMMUNE DUCTS.
BUT THE SKIN ISN'T JUST ON THE
OUTSIDE OF US.
WE HAVE RESPIRATORY EPITHELIA,
GASTROINTESTINAL, REPRO DUCK
TRI, URINARY, AUDITORY, OCULAR,
MUCOSAL SURFACES WHICH HAVE NOT
ONLY UNIQUE EPITHELIAL SURFACES,
EVEN IN THE GASTROINTESTINAL
TRACT, THERE ARE DIFFERENT PATHS
OF THE GASTROINTESTINAL TRACT
WHICH HAVE VASTLY DIFFERENT
DIFFERENTIATION IN THE
EPITHELIUM.
AND ANOTHER PART OF THE HOST
DEFENSE IS THE MICROBIOTA.
I KNOW THAT WAS MENTIONED
EARLIER, BUT THE MICROBIOME IS
VERY EFFECTIVE AT COMPETING OR
DESTROYING POTENTIALLY INVADING
PATHOGENS.
AND I THINK WHEN WE TALK ABOUT
HOST DEFENSES, WE OFTEN DON'T
THINK ABOUT THE MICROBIOME.
AND I'D LIKE TO PROPOSE TO YOU
THAT IN THE CONTEXT OF THE
MICROBIOME, HEALTH IS A
CONDITION OF SYMBIOSIS BETWEEN
THE HOST AND ITS DIVERSE
MICROBIOME, AND THAT DYSBIOSIS
IS A CONDITION IN WHICH THE HOST
IS COMPROMISED BY DISTURBANCE IN
ITS NORMAL MICROBIOME, LEADING
TO AN UNHEALTHY CONDITION,
ILLNESS OR EVEN DEATH.
AND I'M NOT EXAGGERATING WHEN I
SAY THAT, BECAUSE LET'S TAKE,
FOR EXAMPLE, A HEALTHY SYMBIOSIS
IN WHICH YOU INTRODUCE
ANTIBIOTICS FOR A VALID REASON.
A BROAD SPECTRUM ANTIBIOTIC WILL
REDUCE MICROBIOME DIVERSITY IN A
HOST, LEADING THEN TO THE
SUSCEPTIBILITY TO PATHOGENIC OR
PARASITIC ORGANISM SUCH AS
C. DIFFICILE.
C. DIFFICILE CAN THEN INVADE AND
CAUSE A LIFE-THREATENING
DISEASE, AND IN FACT, FURTHER
ANTIBIOTICS MAY ACTUALLY
EXACERBATE THE INFECTION AND
ACCENTUATE THE DYSBIOSIS AND
PROVIDE AN EVEN STRONGER BIAS
FOR C. DIFFICILE TO TAKE HOLD.
AS WE SEE, FECAL GAVAGE CAN
ACTUALLY SAVE LIVES, WHEN DOING
IT FROM HEALTHY VOLUNTEERS AND
SUCCESSFULLY SAVING LIVES AS
MANY CENTERS GLOBALLY HAVE BEEN
DOG.
IN FACT THIS THERAPY, AS CRUDE
AS IT IS, HAS BEEN ACCEPTED BY
REGULATORY AUTHORITIES IN THE
U.K. AND WE'RE NOW THINKING
ABOUT USING IT FOR DISEASES SUCH
AS M.E., AND WE'RE THINKING
ABOUT IT IN TERMS OF AGING AND
TRYING TO PROMOTE HEALTHY AGING
IN A DICE A DYSBIOSIS SITUA.
EVEN PRE-TERM INFANTS GIVEN
MATERNAL FLORA MICROBIOME
INOCULUM, IT'S A LIFE SAVING
TREATMENT AND HAS BEEN SHOWN
HIGHLY SIGNIFICANTLY TO REDUCE
PRE-TERM INFANT DEATHS.
SO THE IMPACT OF A DRUG NEEDS TO
CONSIDER BOTH THE HOST AND ITS
MICROBIOME SYMBIOSIS.
SO THOSE ELEMENTS I'VE JUST BEEN
TALKING ABOUT ARE REALLY VERY
MUCH DIRECTED AT THE FIRST THREE
STEPS IN THE HOST -- IN THE
PATHOGEN INTERACTION WITH THE
HOST, NAMELY CONTACT, ATTACHMENT
AND ADHESION.
BUT TO ADDRESS LATER STAGES IN A
HOST PATHOGEN INTERACTION, WE
NEED TO BECOME -- WE NEED TO
MOVE AWAY FROM INNATE INTO THE
ADAPTIVE PROCESSES.
I'VE TALKED ABOUT AVOIDANCE
WHICH IS BEHAVIORAL, I'VE TALKED
ABOUT PHYSICAL BARRIERS SUCH AS
EPITHELIUM AND CLEARANCE SUCH AS
THE MUCOUS ESCALATOR,
COMPETITION BY THE MICROBIOME.
BUT THE INNATE IMMUNE SYSTEM CAN
CAUSE AN ALARM TO BE INITIATED
AND THOSE ARE BY EXAMPLE TOLL
LIKE RECEPTORS, NATURAL KILLER
CELLS OR T-CELLS, BUT AS WE
BECOME MORE ADAPTIVE, WE CAN
HAVE POLICING FOR MACROPHAGES
LOOKING FOR THE ANTIGEN AND THEN
WE HAVE THE T-CELL IMMUNE
RESPONSE AND DENDRITIC AND NK
CELLS HAVE ALSO A ROLE IN
POLICING.
AS WE BECOME MORE ADAPTIVE,
SPECIFIC CHEMICAL INHIBITION CAN
OCCUR FROM ANTIBODIES THAT ARE
GENERATED BY VACCINE OR PREVIOUS
INFECTION.
AND WE SEE DETECTION OF A
PATHOGEN BY AN ANTIBODY BINDING
TO THE SURFACE OF EITHER A
BACTERIA OR VIRUS, AND THEN
CREATING A FLAG BY THE FC
PORTION OF THE ANTIBODY, WHICH
THEN ENCOURAGES OTHER ELEMENTS
OF THE IMMUNE SYSTEM TO KICK IN.
WE ALSO SEE A SEEK AND DESTROY
FUNCTION FROM OUR ADAPTIVE
IMMUNE RESPONSE, WHERE
ANTIBODIES CAN MEDIATE KILLING
BY COMPLEMENT, NATURAL KILLER
CELLS, DENDRITIC CELLS, AND IN
FACT T-CELLS AND OTHER ELEMENTS
OF THE ADAPTIVE IMMUNE RESPONSE.
SO WE CAN SEE THAT IMMUNITY
PROVIDES US PROTECTION TO SOME
OF THE OTHER STEPS IN A
PATHOGEN-HOST INTERACTION,
INCLUDING INNOVATION AND
REPRODUCTION OF THE PATHOGEN AND
IT MAY EVEN PRESENT
DISSEMINATION OF THE PATHOGEN.
AND SO WE SEE THAT A MODEL THAT
NEEDS TO -- THAT REFLECTS THAT
INTERACTION HAS TO BE EXTREMELY
COMPLEX.
BUT WE STILL HAVE A GAP IN THAT
PATHOGEN INTERACTION.
THE GAP BEING INNOVATION,
TRANSCRIPTION, PIRACY,
INTERFERENCE WITH THE HOST AND
EVEN ASSEMBLY OF THE VIRUS.
DRUGS CAN BE VERY EFFECTIVE AT
DOING THAT.
BUT WHEN I SAY THERAPEUTICS,
THAT'S ALSO INCLUDING SMALL
MOLECULES SUCH AS ANTIBODIES OR
NANOBODIES WHICH OVERLAP INTO
OUR ADAPTIVE IMMUNE SYSTEM.
SO IF WE'RE GOING TO MODEL A
THERAPEUTIC WHICH HAS THAT TYPE
OF COMPLEXITY, WE HAVE TO HAVE A
MODEL THAT'S SUITABLE TO DO SO.
SO WHEN I TALK ABOUT MODEL
SUITABILITY, I WANT TO BEGIN BY
TALKING ABOUT THE INTERACTION OF
A SELECT AGENT WITH THE HOST
CELL.
WHEN WE TALK ABOUT SELECT
AGENTS, THAT DOES INCLUDE
EUKARYOTES, PRO CARE YOATS,
VIRUSES, WHICH IS THE SUBJECT OF
TODAY'S AND TOMORROW'S TALKS,
BUT ALSO TOXINS.
WHEN WE TALK ABOUT ANTIBIOTICS,
WE NATURALLY THINK ABOUT
ANTIBIOTICS TARGETING THE
PROKARYOTES, BUT THEY ACT
DIRECTLY ON THE SELECT AGENT.
SO THOSE DRUGS TARGET THE
REPRODUCTIVE PROCESSES OR
VULNERABILITIES OF BOTH YOU'RE
CARE YOTS AND PROKARYOTICS
DIRECTLY AND THAT KILLS THEM OR
STOPS REPRODUCTION DIRECTLY.
BUT WHEN WE TALK ABOUT AN
ANTIVIRAL DRUG, S, IT NEEDS TO
TARGET THE REPLICATED MECHANISMS
AND IT NEEDS A HIGH FIDELITY
HUMAN HOST MODEL.
IT MAY BE THAT SOME CANCER CELL
LINES OR OTHER CELL LINES MAY
NOT PROIF THIS PROVIDE THF
METABOLIC COMPLEXITY AND THIS IS
WHERE 3D MODELS MAY OFFER THE
COMPLEXITY THAT IS REQUIRED
BECAUSE YOU CAN THEN USE DRUGS
THAT TARGET THE CORRECT
MECHANISMS WITHIN THE HOST CELL
FOR WHICH THE VIRUS IS RELIANT.
I NO VACCINES AREN'T TOPIC TODAY
BUT WE NEED TO CONSIDER THE HOST
IMMUNE SYSTEM IS ABSOLUTELY
PIVOTAL FOR THE ACTION OF A
VACCINE, A VACCINE THAT CAN ONLY
WORK IF THE HOST IMMUNE SYSTEM
RESPONDS TO IT AND IT'S ONLY
THROUGH THE HOST IMMUNE RESPONSE
THAT YOU CAN SEE INHIBITION OF
REPRODUCTION OR KILLING.
SOME THERAPEUTIC DRUGS ACTUALLY
HAVE AN IMMUNE FUNCTION, AND WE
CAN SEE HERE IN THE DIAGRAM THAT
THOSE THERAPEUTIC DRUGS ARE
ACCESSING THE HOST IMMUNE SYSTEM
TO GENERATE THEIR EFFECT ON THE
HUMAN HOST.
SO THERAPEUTIC MONOCLONALS AND
NANOPARTICLES MAY ACT DIRECTLY
OR INDIRECTLY ON THE HOST IMMUNE
SYSTEM.
AN EXAMPLE IS A MONOCLONAL
ANTIBODY MAY ACTUALLY PROVIDE A
LINK TO A NATURAL KILLER CELL TO
TARGET BY THE FC PORTION.
WHAT I WANTED TO DO IS BUILD ON
SOME OF THE POINTS THAT WERE
RAISED IN THE INTRODUCTORY TALKS
AND PROVIDE A QUALITATIVE
ASSESSMENT OF IN VITRO MODELS
ENGAGED IN IN VIVO ALTERNATIVES.
WHEN WE TALK ABOUT A HUMAN
POPULATION, A HUMAN POPULATION
HAS VAST GENETIC DIVERSITY, BUT
WITHIN THAT GENETIC DIVERSITY,
EACH PERSON HAS WITHIN THEM MANY
BODY SYSTEMS AND I'VE LISTED 11
OF THESE BODY SYSTEMS.
AND THAT COMPLEXITY OF THE HOST
POPULATION IS ACTIVELY REFLECTED
IN HUMAN CLINICAL TRIALS, AND IN
HUMAN CHALLENGE STUDIES, AND
IT'S FAIRLY WELL REFLECTED IN
ANIMAL MODELS.
BUT IN VERO CELLS, FOR EXAMPLE,
MANY OF THOSE BODY SYSTEMS ARE
NOT U.S. RT POOED SUPPORT
YOU COULD SAY THE URINARY TRACT
IS SUPPORTED BECAUSE THERE ARE
CELLS, MONKEY KIDNEY CELLS THAT
ARE NOT FULLY DIFFERENTIATED IN
TERMS OF A MONKEY KIDNEY, SO
GIVE IT A PLUS OR MINUS.
MICROPHYSIOLOGICAL SYSTEMS HAVE
REVOLUTIONIZED IN VITRO
TECHNOLOGY AND IT MAY BE THAT
THESE COMPLEX IN VITRO SYSTEMS
CAN ADD SOME SYSTEM
REPRESENTATION BECAUSE YOU CAN
GET MORE THAN ONE TISSUE TYPE
INTO ONE CHIP.
SO FOR EXAMPLE, IN A SINGLE
MICROPHYSIOLOGICAL SYSTEM, YOU
MIGHT HAVE BOTH RESPIRATORY AND
CIRCULATORY CELLS REPRESENTED IN
ONE CHIP, AND IF YOU
INTERCONNECT TWO CHIPS, YOU
MIGHT, FOR EXAMPLE, BE ABLE TO
GET NEURAL, RESPIRATORY AND
CIRCULATORY REPRESENTATION.
SO WE CAN SEE WE'RE GETTING
BETTER BODY SYSTEMS
REPRESENTATION IN THE
MICROPHYSIOLOGICAL SYSTEMS.
IF I START ASSIGNING SINGLE
SCORE POINTS TO EACH TIP AND
BUILDING UP A QUALITATIVE
ASSESSMENT, BUT WHEN WE TALK
ABOUT PROTECTION IN HUMAN
POPULATION, AS I REFERRED TO
EARLIER, WE HAVE BEHAVIORAL
PROTECTION THROUGH AVOIDANCE, WE
HAVE PHYSICAL PROTECTION FROM
THE SKIN, INNATE PROTECTION FROM
IMMUNE SYSTEM, AND ACQUIRED
IMMUNITY.
AND YOU CAN SEE HUMAN CLINICAL
TRIALS ACCURATELY REPRESENT
HUMAN POPULATIONS.
BUT HUMAN CHALLENGE DOES NOT
HAVE THAT BEHAVIORAL AVOIDANCE
THAT THE HUMAN POPULATION HAS,
BECAUSE THE SUBJECTS ARE BEING
DELIBERATELY INFECTED IN THE SAY
WAY ANIMAL MODEL IS, SO WE'RE
USING A SCORE THERE FOR BOTH
HUMAN CHALLENGE AND ANIMAL
MODELS.
IN IN VITRO SYSTEM, ONCE AGAIN,
VERO CELLS MAY REPRESENT INNATE
PROTECTION, BUT THEY DO HAVE
COMPROMISED INTERFERON
CHROMOSOME, WHEREAS IN OUR
MICROPHYSIOLOGICAL SYSTEM, WE'RE
USING CELLS THAT HAVE GOOD
INNATE PROTECTION AND GOOD
PHYSICAL PROTECTION.
SO YOU CAN SEE THE SCORES ARE
SLIGHTLY BUILDING UP THERE ON
MICROPHYSIOLOGICAL SYSTEMS IN
TERMS OF THIS ARBITRARY
SCORECARD.
I WANT TO NOW EXTEND THAT
SCORECARD TO TALK ABOUT
SCIENTIFIC VARIABLES.
WHEN WE TALK ABOUT HUMAN
POPULATIONS, WE HAVE NO IDEA
ABOUT PREVIOUS EXPOSURE, AND
IT'S VERY DIFFICULT TO ESTABLISH
THAT IN HUMAN CLINICAL TRIALS.
CERTAINLY IF YOU HAVE A
VOLUNTEER FOR HUMAN CHALLENGE,
IT'S NOW BECOMING VERY DIFFICULT
TO KNOW WHETHER OR NOT ANY
VOLUNTEER HAS HAD PREVIOUS
INFECTION WITH SARS-COV-2 OR
CROSS-REACTING CORONAVIRUS,
BECAUSE SEROLOGY MAY NOT GIVE
YOU THAT READOUT OF DEEP BONE
MARROW MEMORY IN THE T-CELLS.
WE WON'T KNOW THE EXPOSURE ROUTE
OR THE DOSE THAT PEOPLE HAD
RECEIVED OF A PATHOGEN IN A
HUMAN CLINICAL TRIAL, AND WE
DON'T KNOW THAT IN THE HUMAN
POPULATION.
IN THE HUMAN CHALLENGE, WE DO
KNOW THAT BECAUSE WE GIVE
THEM -- NOT WE, BUT WHOEVER DOES
THAT WOULD GIVE A KNOWN DOSE BY
A KNOWN ROUTE, AND THAT'S TRUE
FOR ANIMAL MODELS, WHERE WE CAN
VERY CAREFULLY ADMINISTER EXACT
DOSES WITH A KNOWN ROUTE.
AND WE CAN COULD THAT IN IN
VITRO SYSTEMS SO ALL THE YESES
GET A GREEN BOX FOR EXTRA POINT.
PREVIOUS EXPOSURE -- WELL
VALIDATED BREEDING FACILITY AND
OF COURSE OUR CELL CULTURES WILL
BE EXPOSURE -- AS WELL.
IN TERMS OF EXISTING IMMUNITY,
I'VE SAID BEFORE IT'S VERY
DIFFICULT TO KNOW WHAT LEVELS OF
IMMUNITY VOLUNTEERS OR TRIAL
PARTICIPANTS HAVE, BUT OF COURSE
ANIMALS OR TISSUE CULTURE WILL
NOT HAVE ANY EXISTING IMMUNITY.
THERE IS GREAT GENETIC
VARIABILITY AS DISCUSSED EARLIER
IN POPULATIONS AND IN CLINICAL
TRIALS, THAT CAN BE WELL
REPRESENTED, AND IN A HUMAN
CHALLENGE, THAT CAN ALSO BE
REPRESENTED.
OBVIOUSLY IN ANIMAL MODEL, THEY
TEND TO BE INNATE IN BRED
ANIMALS BUT OF COURSE YOU CAN
SUES DIFFERENT HAPLOTYPES WITHIN
ANIMAL MODELS TO REPLICATE
WHAT'S HAPPENING IN THE HUMAN
CHALLENGE.
OBVIOUSLY WE COULD TAKE
REPRESENTATIVES AND GET
DONATIONS FROM DIFFERENT GENETIC
GROUPS SO THOSE COULD BE YESES.
IN TERMS OF UNDERLYING
CONDITIONS, IN HUMAN CLINICAL
TRIALS, THERE CAN BE MANY
UNDERLYING CONDITIONS BUT IN A
HUMAN CLINICAL TRIAL, THERE
WON'T BE ANY BECAUSE WE CAN
SELECT THOSE CLINICAL UNDERLYING
CONDITIONS OUT AND THAT'S THE
SAME IN ANIMAL MODELS AND
MICROPHYSIOLOGICAL SYSTEMS,
BECAUSE IN VERO CELLS, THERE IS
EFFECTIVELY AN UNDERLYING
CONDITION BECAUSE THERE IS NO
INTERFERON PRODUCTION.
IN TERMS OF RISK OF HARM, THAT
WAS MENTIONED EARLIER IN A HUMAN
POPULATION, THERE'S LOW RISK OF
HARM.
IN A HUMAN CLINICAL TRIAL,
THERE'S MODERATE RISK OF HARM
BECAUSE THAT PLACEBO IS NOT
PROVIDING PROTECTION OF THE
VACCINE UNLESS YOU MITIGATE
THAT.
AND CERTAINLY IF YOU WANT TO DO
A HUMAN CHALLENGE WITH A SELECT
AGENT, THERE'S A HIGH RISK OF
HARM HERE.
IN TERMS OF ANIMALS, THERE'S A
MODERATE RISK OF HARM, BOTH ET
CAL AND IN TERMS OF SUFFERING,
BUT IN TERMS OF IN VITRO AND
MICROPHYSIOLOGICAL SYSTEMS, THE
RISK OF HARM IS EXTREMELY LOW,
HANDLED IN A CATEGORY
3 LABORATORY.
AND OF COURSE ANIMAL MODELS ARE
NOT HUMAN, AND VERO CELLS AREN'T
HUMAN EITHER, BUT
MICROPHYSIOLOGICAL SYSTEMS GIVE
YOU A HUMAN REPRESENTATION IN
THE WAY THAT THE OTHER IN VITRO
SYSTEMS DON'T.
SO IF I ADD ALL OF THESE
QUALITATIVE POINTS TOGETHER, YOU
CAN SEE THAT THE
MICROPHYSIOLOGICALS' SCORE HAS
ACTUALLY CREPT UP QUITE WELL.
AND I HAVEN'T DONE THIS YET, BUT
IF WE ADD IN ACQUIRED IMMUNITY
INTO MICROPHYSIOLOGICAL SYSTEMS,
I THINK THEIR QUALITATIVE SCORE
WILL POSSIBLY REACH OR EXCEED
THAT OF THESE OTHER TYPES OF
HUMAN CLINICAL TRIAL.
SO NOW I WANT TO TALK ABOUT
TOXICOLOGY.
I KNOW IT WAS MENTIONED EARLIER
AS WELL.
TOXICOLOGY IS IN SOME RESPECTS A
FAIRLY SIMPLE SCIENTIFIC ASPECT
OF BIOLOGICAL SCIENCE, BECAUSE
IT INVOLVES CELL CULTURE, CELL
BIOLOGY, MOLECULAR BIOLOGY,
THERE'S BEAUTIFUL ROBOTICS AND
MICRO FLUIDICS NA HAVE THATN
ADVANCED IN THE LAST DECADE, AND
THOSE PK AND PD STUDIES ARE VERY
PRECISE AND ACCURATE.
AND ACTUALLY HAVE BEEN VERY
ILLUMINATING FOR REGULATORS.
BUT WHEN WE TALK ABOUT CANCER
RESEARCH, THERE'S ANOTHER ORDER
OF COMPLEXITY IN THOSE STUDIES.
BECAUSE NOW YOU INCLUDE
IMMUNOHISTO PATHOLOGY, HUMAN
IN SITU HYBRIDIZATION,
BIOLUMINESCENCE, POSSIBLY
TRANSCRIPTOMICS AND PROTEOMICS.
BUT CANCER RESEARCH IS ALSO
PERHAPS LESS COMPLICATED THAN
VIROLOGY, BECAUSE VIROLOGY HAS
ALL OF THE COMPONENTS OF CANCER
RESEARCH ON TOP OF THE
REQUIREMENTS TO HANDLE BSL LEVEL
3 PATHOGENS AND IT REQUIRES
ILLUMINATION AND TITRATION OF
THOSE PATHOGENS, SEROLOGY
AGAINST THE PATHOGEN AND
IMMUNOLOGY AGAINST THE PATHOGEN
OF INTEREST.
BUT IT DOES HAVE MANY OF THE
FEATURES OF THE TWO OTHER
FACULTIES OF BIOLOGICAL SCIENCE.
I WAS SURPRISED AT THIS AND I
WANTED TO DRAW TO EVERYBODY'S
ATTENTION, THAT THERE IS
REGULATORY ADAPTATION OCCURRING
AS WE SPEAK IN THE FDA
MODERNIZATION ACT OF 2021, HERE
ON THE LEFT-HAND SIDE, THE
PHRASE THEY WANT TO USE
ALTERNATIVE TESTING METHODS TO
ANIMAL TESTING TO INVESTIGATE
THE SAFETY AND EFFECTIVENESS OF
A DRUG AND FOR OTHER PURPOSES.
AND THIS HAS BEEN SUPPORTED BY
JIM CORBETT AND MANY OTHER
PEOPLE, SUGGESTING THAT SOME OF
THE AP MALL ANIMAL STUDIES E
OCCURRED HAVE LED TO THE WRONG
CONCLUSION WHEN IT COMES TO
HUMANS.
SO WHAT WE'RE SEEING HERE IS A
PARADIGM SHIFT IN REGULATORY
ACCEPTANCE OF MICROPHYSIOLOGICAL
SYSTEMS.
INTO AND I CAN I I CASSIS
IN TOXICOLOGY AND CANCER
RESEARCH WILL SPREAD THROUGH
INTO MICROBIOLOGY AND IMMUNOLOGY
QUITE RAPIDLY, BECAUSE THAT
CROSS-CUTTING ADVANCES ARE GOING
TO IMMEDIATELY HAVE AN IMPACT ON
REGULATORS AND FUNDERS AND
DEVELOPERS.
I'VE TRIED TO DRAW A SIMPLE
REPRESENTATION OF
MICROPHYSIOLOGICAL SYSTEMS AND
THE ADVANTAGES THEY OFFER AS
WELL.
BECAUSE FOR EXAMPLE, A
TWO-TISSUE MICROPHYSIOLOGICAL
SYSTEM HAS TWO CHAMBERS, WE CAN
MODEL SYSTEMIC DOSING OF
ANTIVIRALS, DRUGS AND
ANTIBIOTICS INTO THE VASCULAR
COMPONENT AND THEN INTRODUCE THE
SELECT AGENT INTO THE ORGAN CELL
CHANNEL OF WHICH MIGHT BE THE
BRAIN, KIDNEY OR OTHER ORGAN.
SO HERE WE'RE ASSIMILATING
SYSTEMIC DOSING.
BUT IN THE SAME CHIP, WE MIGHT
HAVE A MODEL MUCOSAL INFECTION
OR DOSING BY APPLYING ANTIVIRAL
DRUG, ANTIBIOTIC OR EVEN IMMUNE
CELLS INTO THE MUCOSAL SURFACE,
ALONG WITH A SELECT AGENT.
SO WE HAVE IMMEDIATELY USED IT
FOR MORE THAN ONE PURPOSE
WITHOUT CHANGING ANYTHING, THE
ROUTE OF ENTRY OF THE DRUG
ANTIVIRAL OR IMMUNE CELLS.
BUT WE CAN ALSO MODEL
COMBINATIONS OF THE TWO, PERHAPS
INTRAVENOUS ANTIVIRAL DRUG
ADMINISTRATION, AS WELL AS
MUCOSAL ANTIBODY OR IMMUNE CELL
ADDITION, AS WELL AS THE SELECT
AGENT.
AND CULMINATIONS THERE.
NOW I WANT TO TALK ABOUT SOME
OLD EXAMPLES AND IN THE EARLY
90s, WHEN I WAS DOING MY PH.D.
ON B. PERTUSSIS, MECHANISMS OF
COLONIZATION OF MAMMALIAN
TISSUES BY BOARD AT THE LA
PERTUSSIS.
IT DOES STICK VERY WELL TO VERO
CELLS BUT IT DOES SO USING
COMPONENTS OTHER THAN THE
SPECIFIC ADHESION.
YOU CAN SEE THIS COMPLEX
CILIATED EPITHELIUM SO CRITICAL
TO THE FIRST STEPS OF CONTACT
BETWEEN BORDETELLA PER PERTUSD
THE CELL.
-- INITIALLY HAS AN ADD ADHEN
EVENT WHICH OCCURS FROM THE END
OF THE -- TO THE CILIATED
EPITHELIUM.
AND THEN THIS LEADS TO BETTER
ADHESION OF THE ORGANISM
FOLLOWING THAT INITIAL
ATTACHMENT THROUGH OTHER
MECHANISMS.
SO THIS LOVELY CILIATED
EPITHELIAL TISSUE MODEL WAS ABLE
TO DEMONSTRATE MANY OF THE
PATHOGEN INTERACTIONS WITH
BORDETELLA PERTUSSIS.
SO IN TERMS OF CON TAJT COS
INITIAL INHALATION FROM AN
AEROSOL FROM A CHILD WITH
WHOOPING COUGH, AND ADD HEE SIN
SPECIFIC FOR HUMAN CILIA AT THE
TIP OF FIMBRIA, OTHER SURFACE
ADHESIVES OF BORDETELLA
PERTUSSIS CAN ADHERE VERY
EFFECTIVELY TO THE SURFACE OF
THE CELL, AND THEN START
INTERFERING WITH THE HOST BY
DISRUPTING THE CILIATED
EPITHELIUM THROUGH TOXINS.
THEN WHEN WE SEE DISSEMINATION
OF DISEASE, IT'S THROUGH AN
EXTREME COUGH, IF YOU'VE EVER
SEE IT, IT'S VERY DISTRESSING,
WHICH
PERTUSSIS TOXIN, WHICH CAN LEAD
TO DISSEMINATION TO THE NEXT
HOST.
EVEN WHEN SOMEONE HAS ELIMINATED
THE ORGANISM FROM THEIR SYSTEM,
THE DISEASE CONTINUES FOR A LONG
TIME, AND THAT'S DUE TO THE
TOXIN REMAINING IN CELLS AND
CAUSING THAT HORRIBLE PAROXYSMAL
COUGH.
SO YOU SEE THE BEAUTY OF THAT
CILIATED EPITHELIUM IS ABLE TO
REFLECT THOSE COMPLEX PROCESSES
LISTED ON THE LEFT-HAND SIDE.
BETWEEN THE 1960s AND 1980s,
THE WIDELY ACCEPTED BORDETELLA
PERTUSSIS WAS -- VACCINE WHICH
WAS INDEED VERY COMPLEX BECAUSE
IT WAS BASICALLY A FORMALDEHYDE
SOUP OF BORDETELLA PERTUSSIS.
IN THE 80s AND EARLY 90s,
THAT VACCINE WAS REPLACED BY ONE
THAT HAD A SINGLE, DOUBLE OR
TRIPLE ANTIGEN COMPONENTS, WHICH
IS ACTUALLY A VERY SIMPLE
VACCINE.
BUT DUE TO VACCINE ESCAPE IN THE
21ST CENTURY, WE'VE HAD TO
INCREASE THE NUMBER OF
COMPONENTS IN EACH VACCINE SO
THEY'RE MUCH MORE COMPLEX, LESS
SIMPLE.
BUT AS WE SPEAK, NOW IN
DEVELOPMENT, THERE IS NOW A LIVE
ATTENUATED B. PERTUSSIS VACCINE
ONGOING ASSESSMENT, AND OF
COURSE THIS VACCINE IS VERY
COMPLEX IN THE SAME WAY THAT WE
HAD IN THE 60s AND 80s, SO
WE'VE GONE FULL LOOP FROM A VERY
COMPLEX VACCINE, FROM A SIMPLE,
BACK TO A VERY COMPLEX.
BUT WHEN WE LOOK AT THE ANIMAL
MODELS AVAILABLE TO MIMIC
BORDETELLA PERTUSSIS, THERE'S
ONLY THE BOONS THAT ACTUALLY
COUGH WHEN THEY'RE INFECTED.
AND SO AT THE MOMENT WE'RE NOW
SEEING HUMAN CHALLENGE MODEL
BEING UTILIZED THE ASSESSMENT OF
A LIVE AT10 WAITED BORDETELLA
PERTUSSIS IN 2022, AND I REALLY
THINK THE HUMAN
MICROPHYSIOLOGICAL SYSTEMS COULD
HAVE ASSISTED IN THIS PROCESS IF
WE HAD THIS WONDERFUL TECHNOLOGY
WHEN I STARTED MY PH.D.
SORE LET'S TAKE ANOTHER EXAMPLE,
POX VIRUSES AND HOW TOPICAL IS
THAT?
ANTIVIRAL DRUGS FOR SMALLPOX
WERE DEVELOPED USING MONKEYPOX
AND THEY CAN BE USED AGAINST
MONKEYPOX.
AND WE TAKE TWO KRUTION DRUE
BEEN APPROVED BY THE FDA, WE CAN
SEE THEY WERE APPROVED IN 2018
AND 2021 RESPECTIVELY.
NOW THE ISSUE WITH SMALLPOX IS
THAT IT'S A HUMAN-SPECIFIC VIRAL
PATHOGEN, SO IT WASN'T EASY TO
MODEL THAT IN ORDER TO DEVELOP
THESE DRUGS AND ACTUALLY THE
VACCINES.
SO MONKEYPOX WAS USED AS A
SURROGATE FOR SMALLPOX IN
NON-HUMAN PRIMATE, AND INDEED IN
U.K. AND MANY OTHER PLACE,
VACCINES WERE TESTED IN NHP
MODELS USING MONKEYPOX.
BUT ONE OF THE OTHER ISSUES WAS
THAT THE FAR KA COKINETICS OF
TEMBAXA IN NHPs WAS VERY
DIFFERENT FROM THAT IN HUMANS.
THAT WASN'T AN EXPECTED OUTCOME.
THAT LED TO -- FDA APPROVAL
GRANTING AND BECAUSE THEY HAD TO
FIND ANOTHER WAY OF ASSESSING
IT, AND IN FACT, VACCINIA CAN
ALSO BE USED AS A SURROGATE FOR
SMALLPOX IN HUMANS.
MAYBE THERE'S A LESSON TO BE
LEARNED THERE BECAUSE MAYBE 3D
MICROPHYSIOLOGICAL SYSTEMS COULD
HAVE BEEN USED IF WE HAD THOSE
TOOLS 15 YEARS AGO WHEN DRUG
DEVELOPMENT WAS ONGOING.
WHEN WE TALK ABOUT EBOLA, THERE
ARE MANY FILOVIRUSES APART FROM
EBOLA AND SOME OF THEM DON'T
INFECT NHPs, SO THIS
REPRESENTS A BLIND ALLEY FOR
THOSE PARTICULAR FILOVIRUS
FAMILY.
WE NOW KNOW EBOLA CAN SEQUESTER
IN EYE, BRAIN AND REPRODUCTIVE
TISSUES.
HOW CAN WE STUDY CLEARANCE FROM
THESE IMMUNE PRIVILEGE SITES
WITHOUT ACTUALLY GOING INTO
INDEMIC AREAS, EVEN IF YOU CAN
DO THAT?
IT'S EXTREMELY DIFFICULT TO TAKE
SAMPLES FROM THOSE IMMUNE
PRIVILEGED SITES.
AND HOW DO WE TEST COMBINATIONS
OF ANTIVIRALS OUTSIDE OF
OUTBREAKS?
ONCE AGAIN, HUMAN
MICROPHYSIOLOGICAL SYSTEMS COULD
ASSIST.
AND WHAT ABOUT HEPATITIS?
THIS IS ALSO A VERY TOUGH
TOPICAL TOPIC BECAUSE WE'RE
SEEING SEVERE CASES IN VERY
YOUNG CHILDREN.
IN THE U.K., THERE ARE MANY
THEORIES ABOUT WHY THIS IS
OCCURRING.
AT THE MOMENT, IT COULD BE DO TO
A NEW ADENOVIRUS OR COULD BE A
NEW VARIANT OF ADENOVIRUS OR IT
COULD BE SOMETHING TO DO WITH A
NEW VARIANT OF SARS-COV-2 OR
COULD BE RELATED TO CO-INFECTION
WITH SARS-COV-2.
AND THERE ARE MANY QUESTIONS
REMAINING ABOUT WHY THERE ARE SO
MANY GLOBAL CASES OF JUVENILE
HEPATITIS, AND IN SOME CASES
REQUIRING LIVER TRANSPLANT,
WHICH IS A LIFE-CHANGING IMPACT.
AND MAYBE HUMAN
MICROPHYSIOLOGICALS COULD HAVE
ASSISTED IN THIS INITIAL
OUTBREAK INVESTIGATION, BUT I
WOULD SAY THAT IT COULD
CONTINUE, MICROPHYSIOLOGICAL
SYSTEMS COULD ASSIST IN
ANSWERING SOME OF THESE
QUESTIONS WHICH ARE VERY TOPICAL
EVEN AS I SPEAK.
SO LET'S TALK ABOUT DISEASE X
BECAUSE THAT ONE IS QUITE A
TOPICAL SUBJECT.
DISEASE X IS THE UNKNOWN AGENT
WHICH WE CAN PREDICT MIGHT BE
THE CASE AND I KNOW SOME OF THE
INTRODUCTORY SPEAKERS HAVE
SPOKEN ABOUT DISEASE X.
BUT LET'S TAKE SCENARIO H WITHIN
THE DISEASE X SCENARIO.
LET'S SAY THAT THE SCENARIO H IS
A VIRUS WHICH ONLY AFFECTS HUMAN
LIVER.
AND FOR EXAMPLE THAT'S FATAL.
NO ANIMAL MODEL IS POSSIBLE
WITHOUT GENETIC TRANSFORMATION
AND WITHOUT A 100-DAY CHALLENGE,
WE'RE NOT GOING TO BE ABLE TO DO
THAT FAST ENOUGH.
A HUMAN INFECTION WITH DISEASE X
SCENARIO H WOULD NOT BE
ETHICALLY JUSTIFIED SO WE'RE
HEAVILY RELIANT ON GENETICS IN
THE INITIAL STAGES OF A
CHALLENGE.
HOW COULD WE TEST DRUGS,
THERAPEUTICS AND VACCINES FOR
SUCH A DISEASE H SCENARIO H?
HERE, I THINK MICROPHYSIOLOGICAL
SYSTEMS COULD COME INTO THEIR
OWN AND ASSIST QUITE
DRAMATICALLY.
AND I THINK WE NEED TO GET THEM
READY FOR THAT SCENARIO, BECAUSE
IT'S THE WORST CASE.
IF WE'RE READY FOR THE WORST
CASE, WE'RE READY FOR ANYTHING.
SO NOW LET ME TALK ABOUT VERO
CELLS, AS MANY OF YOU WILL KNOW
THEY'RE THE VIROLOGIST'S
FAVORITE BECAUSE THEY'RE
AVAILABLE GLOBALLY, THEY GROW
MANY VIRUSES VERY EASILY,
INCLUDING SARS-COV-2 AND THEY'RE
WIDELY USED FOR VIRAL
PROPAGATION.
THEY DO HAVE AS MENTIONED
EARLIER A DELUSION IN CHROMOSOME
12, WHICH RESULTS IN THE LOSS OF
ABILITY TO PRODUCE TYPE
1 INTERFERON.
THERE'S ALSO A LOSS OF SOME
ELEMENTS OF REGULATION AND
THAT'S WHY THEY GROW SO WELL,
AND THAT'S REPRESENTED IN OTHER
DELETIONS IN THE CHROMOSOME
WHICH YOU'LL READ ABOUT IN 2014.
THAT'S NOT ONLY FROM KIDNEY
ORIGIN BUT ALSO FROM A GREY
MONKEY.
AND I CAN'T HELP MENTIONING MY
OWN ARTICLE WHICH I PUBLISHED
WITH THE ASSISTANCE OF MANY
PEOPLE WHO MAY ALSO BE ON THE
LINE TODAY, WHICH WAS A
CAUTIONARY PERSPECTIVE ABOUT
ISOLATION OF SARS-COV-2 2 AND
THE URGE FOR PEOPLE TO CHECK
ANYTHING THEY GROW IN VERO CELLS
BECAUSE WE CAN SEE GENETIC
DIVERGENCE WITHIN THESE CELLS
THAT DON'T HAVE INTERFERON
PRODUCTION.
IF YOU DO USE A STOCK WHICH IS
COMPROMISED, IT WILL RESULT IN
REDUCED PATHOGENICITY IN AN
ANIMAL MODEL, LOSS OF INTERFERON
CLEAVAGE SITE.
THESE ARE TABLE FROM THE PAPER
WHICH YOU CAN LOOK AT LATER.
IT'S NOT JUST ABOUT THE SPIKE.
THERE ARE OTHER CHANGES THAT
OCCUR IN SOME CULTURES.
IN THIS PARTICULAR, WE SAW IT I
IN -- WE STILL DON'T REALLY KNOW
ABOUT.
ALSO WANT TO MENTION A PAPER
WHICH WE PRODUCED ABOUT
HYDROXYCHLOROQUINE WHICH IS
REALLY -- THAT IS THAT VERO CELL
DATA WE HAD ON HAND AT THE
BEGINNING OF THE OUTBREAK SEEMED
TO SUGGEST THAT
HYDROXYCHLOROQUINE HAD ANTIVIRAL
AND SYMPTOMATIC IMPACT IN THE
VERO CELL MODEL OF INFECTION.
BUT AS A RESULT OF COLLABORATION
WITH THE WHO, WHEN WE ASKED
DIFFERENT AUTHORITIES IF THEY
HAD HYDROXYCHLOROQUINE DATA,
EACH LABORATORY HAD A LITTLE BIT
OF INFORMATION AND WHEN YOU
PIECED IT TOGETHER, EACH PIECE
OF THE JIGSAW SEEMED TO SUGGEST
THERE WAS NO ANTIVIRAL
SYMPTOMATIC RELIEF IN ANY ANIMAL
MODEL AVAILABLE AT THE TIME.
BUT THE MOST INTERESTING THING
FOR THIS AUDIENCE TODAY IS THAT
TWO INDEPENDENT LABORATORIES
USING MICROPHYSIOLOGICAL SYSTEMS
ALSO SUPPORTED THE IN VIVO
OUTCOMES, WHICH IS THAT
HYDROXYCHLOROQUINE HAD NO
ANTIVIRAL OR SYMPTOMATIC IMPACT.
AND THIS PAPER POSSIBLY ASSISTED
IN THE LACK OF UTILIZATION OF
HYDROXYCHLOROQUINE FROM THAT
POINT ON WARDS.
SO MICROPHYSIOLOGICAL SYSTEMS OF
COURSE HAVE ALREADY PROVEN THEIR
WORTH, NOT ONLY FOR HYDROXYCOLOR
QUICK BUT ALSO FOR IMATINIB,
WHICH HAS A VERY SIMILAR STORY.
I KNOW WE'RE TALKING ABOUT
ANTIVIRALS, BUT LET ME TALK
ABOUT HUME RORL ORAL IMMUNITE
CONTEXT OF SARS-COV-2.
THAT CARTOON OF VIRUS ENTRY AND
DISSEMINATION ILLUSTRATES SOME
OF THE KIT CAL PARTS OF VIRUS
REPLICATION.
WHEN WE TALK ABOUT HUMAN
ANTIBODIES, WE OFTEN TALK ABOUT
THE FIRST TWO OF FIVE
CONSEQUENCES OF ANTIBODY
BINDING, WHICH IS INHIBITION OF
BINDING OF THE VIRUS TO THE
SURFACE OF THE HOST CELL BUT
ALSO INHIBITION OF FUSION OF THE
VIRUS WHICH SUGGESTS UP TAKE
INTO THE CELL.
AND THOSE TWO STEPS ARE VERY
WELL REPRESENTED IN A
NEUTRALIZATION ASSAY, AND WE'VE
BEEN PRODUCING MICRO
NEUTRALIZATION ASSAY AGAINST ALL
THE VARIANTS OF CONSENT SINCE
VERY EARLY ON IN THE OUTBREAK.
AND AS WE CAN SEE OVER TIME,
WHEN WE LOOK AT THE VARIANTS OF
CONCERN COMPARED TO VICTORIA,
OMICRON WAS PREDICTED TO HAVE
VERY LITTLE NEUTRALIZATION
CAPABILITY IN TERMS OF HUMORAL
IMMUNITY.
AS THE OTHER SPEAKERS HAVE
MENTIONED, IT WASN'T JUST ABOUT
APT BO DIES.
ANTIS.
WHEN WE ACTUALLY USED OMICRON TO
INFECT HAMSTERS, IT PRODUCED A
VERY MILD FORM WHICH YOU CAN SEE
ON THE LEFT.
WE SAW SOME -- SIGNS AND WE DID
SEE A LEVELING OUT RATHER THAN
REDUCTION IN WEIGHT.
BUT WHEN WE TOOK HAMSTERS THAT
HAD ACTUALLY RECOVERED FROM
VICTORIA, THE SEROLOGY WOULD
HAVE PREDICTED THAT THAT HUMORAL
RESPONSE WOULD NOT HAVE -- OUT,
BUT IN FACT CONVALESCENT
HAMSTERS WERE ACTUALLY VERY WELL
PROTECTED AGAINST OMICRON.
SO WHY WOULD CONVALESCENT
HAMSTERS BE CROSS-PROTECTED WHEN
THE NEUTRALIZATION ASSAYS
PREDICTED THAT THEY WOULDN'T BE?
AND IT COULD BE THAT THE HAMSTER
ANTIBODIES WERE MUCH STRONGER
AND MORE LASTING THAN IN HUMANS,
BUT IT'S MORE LIKELY THAT THE
HUMAN -- THE HAMSTER INFECTION
MODEL INCLUDES T-CELL IMMUNITY
AND THE CD4 AND CD8 CELL
COMPONENTS OF THE IMMUNE
RESPONSE WAS RESPONSIBLE IN PART
FOR THAT CROSS-PROTECTION.
IN ADDITION, THE HAMSTER MODEL
INCLUDES ANTIBODIES, INDUCING A
COMPLEMENT CASCADE, WHICH MAY
NOT BE ACCURATELY REFLECTED IN A
MICRO NEUTRALIZATION ASSAY.
AND ANOTHER ALTERNATIVE
CONTRIBUTOR COULD BE THAT THE
HAMSTER INCLUDES ANTIBODY
TARGETED CELLULAR COMPONENTS.
BY THAT, I MEAN ANTIBODIES
BRIDGING FC RECEPTORS TO
DENDRITIC AND MACROPHAGE CELLS,
AND ALSO CYTOTOXIC KILLER CELLS
BEING FLAGGED, SUCH AS NATURAL
KILLER CELLS TO THE SURFACE OF
INFECTED CELLS.
SO WHEN WE TALK ABOUT MODELS OF
THE INTERACTION BETWEEN HOST AND
PATHOGEN IN AN IMMUNE CONTEXT,
WE SHOULD BE THINKING ABOUT HOW
WE CAN INCLUDE CELLULAR IMMUNITY
INTO THESE MODELS, AND YOU CAN
SEE HERE, CELLULAR IMMUNITY HAS
QUITE A DIVERSE ARRAY OF
CHARACTERS AS ILLUSTRATED IN
THIS FROM THE BBC WEBSITE.
SO THE THREE OTHER ELEMENTS OF
ANTIBODY BINDING CONSEQUENCES
ARE THAT AN ANTIBODY CAN
ACTUALLY PRODUCE A COMPLEMENT
CASCADE OR VIRAL OPSONO
PHAGOCYTOSIS BY FLAGGING --
EXCUSE ME
[KNOWN
[PHONE RINGING]
-- BY FLAGGING VIRAL OPSONO
PHAGOCYTOSIS AND ALSO BY
FLAGGING CELLULAR ELEMENTS OF
THE IMMUNE SYSTEM SUCH AS
NATURAL KILLER CELLS,
MACROPHAGES OR LOCAL DENDRITIC
CELLS BY THE SE COMPONENTS.
BUT IN THAT DATA THAT I'VE JUST
SHOWN, LET'S NOT FORGET ZETA.
I KNOW IT'S GONE, BUT IT DOESN'T
APPEAR TO BE DETECTED ANYWHERE,
BUT THE STORY ABOUT ZETA IS VERY
INTERESTING BECAUSE UNLIKE
OMICRON, IT ONLY HAD ONE CHANGE
IN THE SPIKE AND OMICRON HAD
MANY CHANGES IN ITS SPIKE.
BUT ZETA HAD A CHANGE IN OH --
MAYBE THERE'S CHANGES IN OTHER
ASPECTS OF THE VIRUS WERE
RESPONSIBLE FOR THE ESCAPE IN
THE MICRO NEUTRALIZATION ASSAY.
MAYBE WE NEED TO INVESTIGATE
THIS IN A MICROPHYSIOLOGICAL
SYSTEM TO UNDERSTAND WHAT WAS
HAPPENING THERE.
WHEN WE LOOK AT A PSEUDOVIRUS
ASSAY, WE DON'T SEE THAT TYPE OF
REDUCTION IN SIGNAL AND THAT MAY
BE BECAUSE THE
PSEUDONEUTRALIZATION ASSAYS ARE
ONLY FOCUSED ON THE SPIKE
PROTEIN.
WHEN WE TALK ABOUT RESEARCH AND
DEVELOPMENT, IN THE CONTEXT OF
SCIENTIFIC SERVICES, I'M
STARTING TO USE THIS DIAGRAM TO
EXPLAIN TO PEOPLE THE DIFFERENCE
BETWEEN RESEARCH AND DEVELOPMENT
IN THIS CONTEXT.
I KNOW IN TERMS OF RESEARCH, WE
TEND TO HAVE HYPOTHESIS TESTING
WHERE WE MOVE FROM AN UNKNOWN
SITUATION TO A KNOWN SITUATION
BY USING A SCIENTIFIC SERVICE TO
ANSWER A SPECIFIC QUESTION.
THE KNOWLEDGE THAT WE GAIN FROM
THAT GIVES US APPLIED KNOWLEDGE,
AND THAT IN ITSELF IS A SIGN OF
SCIENTIFIC SERVICE.
BUT MAKING SURE THAT OUR
SCIENTIFIC SERVICES REMAIN ON
TARGET IS AN IMPORTANT COMPONENT
OF A SCIENTIFIC SERVICE, AND A
QUALITY MANAGEMENT SYSTEM WILL
DETECT WHETHER OUR SCIENTIFIC
SERVICES ARE OFF TARGET OR NOT.
AND IF THEY ARE, WE THEN NEED TO
APPLY DEVELOPMENT TO RE-TARGET
OUR SCIENTIFIC SERVICES IN ORDER
TO IMPROVE OUR SERVICE TO ENSURE
WE SEE THE WHOLE PICTURE, NOT
THE TIP OF THE ICEBERG.
THIS IS EXACTLY WHAT HAPPENED
WITH SARS-COV-2 DURING OWRL
RESPONSE TO THE PANDEMIC,
BECAUSE OF THE EMERGENCE OF
VARIANTS OF CONCERN.
WHEN WE TALK ABOUT AN ANIMAL
MODEL WE HAD AVAILABLE, THESE
WERE VERY WELL ILLUSTRATED IN
MANY COLLABORATORS, WE SAW THERE
WAS FOR EACH VARIANT OF CONCERN,
THERE WAS A SLIGHTLY DIFFERENT
PROFILE OF INFECTIVITY BETWEEN
HAMSTERS, PRIMATES, FERRETS AND
SMALL ANIMALS.
BUT THIS HOST RANGE VAIR VARS
SOMETHING WE JUST CAN'T DEAL
WITH IN TERMS OF ANIMAL MODELS.
SO ASSAYS NEED TO BE FUTURE
PROOFED, AND IF SARS-COV-2 IS
EVOLVING INTO HUMAN-ONLY FORM,
MAYBE THESE ANIMAL MODELS WILL
EFFICIENTLY BECOME REDUNDANT.
UNLESS WE USE RECOMBINANT
ANIMALS.
BUT THIS IS NOT THE LONG TERM
ANSWER, AND I THINK THE
MICROPHYSIOLOGICAL SYSTEMS MAY
OFFER A FUTURE PATHWAY TO NOT
ONLY VARIANTS OF CONCERN BUT
OTHER HUMAN SPECIFIC PATHOGENS.
AS WE CAN SEE IN THIS PICTURE
FROM LONGLONG.
SO IN THE LAST FEW MINUTES, I
JUST WANTED TO TALK ABOUT SUPPLY
AND DEMAND.
WE DO NEED A PLAN.
FOR SMALL ANIMAL STUDIES, THAT'S
FINE, BUT TRANSGENICS TOOK A LOT
OF MONEY AND THEY TOOK TIME TO
DEVELOP, AND ACTUALLY WHEN WE
DID HAVE SOME OF THEM, THEY WERE
EXTREMELY SEVERE FORM OF
INFECTION WHICH DIDN'T REPRESENT
A NORMAL FORM OF INFECTION.
FOR NON-HUMAN PRIMATE, IT'S NOT
OKAY TO LOOK AT EACH VARIANT
CONCERN FOR EFFECTORS OF
VACCINES AND DRUGS BECAUSE
THERE'S SIMPLY NOT ENOUGH
ANIMALS AND WE CAN'T COPE WITH
ALL THE VARIANTS OF CONCERN,
EVEN IF WE WANTED TO, AND IT MAY
BE THAT IT WILL DRIFT AWAY FROM
HAVING SPECIFICITY TO NHPs.
SO IN MICROPHYSIOLOGICAL SYSTEM,
THEY CAN BE USED FOR CELL LINES
BUT THEY'RE NOT REPRESENTATIVE
OF COMPLEX TISSUES BUT COULD USE
HUMAN PRIMARY TISSUE BUT THERE'S
VARIANCE IN ETHICAL CONCERNS AND
WE NEED TO HAVE SOMETHING THAT'S
RELIABLE AND REPRODUCIBLE OVER
TIME.
SO MAYBE POTENTIAL PLEUR OWE
POTENT STEM CELLS IN THE FUTURE
AND IF SO WE NEED TO THINK OF A
GLOBAL SUPPLY AND ALTHOUGH THAT
MIGHT BE EXPENSIVE IN THE FIRST
INSTANCE, DEMAND AND SCALE WILL
REDUCE THAT COST.
SO IN CONCLUSION, WELCOME TO THE
21ST CENTURY SCIENTIFIC
RENAISSANCE.
THE GLOBAL COMMUNITY IS NOW
LISTENING TO US.
WE'RE NOW SHARING DATA AND
TECHNOLOGY IN A WAY WHICH WE
COULDN'T HAVE ANTICIPATED AND
THIS IS NOW THE NEW NORM.
I THINK THE HUMAN
MICROPHYSIOLOGICAL SYSTEMS
OFFERING A POTENTIAL TO ANIMALS
IN THE LONG TERM, BUT WE NEED TO
FIT THOSE ANTIVIRAL DRUG TESTING
REQUIREMENTS, BUT THOSE THAT
TICK THE BOXES FOR ANTIVIRAL
DRUG TESTING SHOULD ALSO BE
FIT-FOR-PURPOSES FOR VACCINES
AND THERAPEUTICS, WHICH COULD BE
INCORPORATED INTO DRUGS.
SO LET'S MAKE SURE THAT THE
MODELS WE CHOOSE ARE APPLICABLE
TO VACCINE AND THERAPEUTIC
TESTING.
INTO AND AND ONCE WE HAVE TH,
THAT COULD BE USED FOR ALL
SELECT AGENTS ONCE WE ADDRESS
ISSUES AROUND CONTAINMENT, WHICH
ARE QUITE FORMIDABLE, BUT WE
NEED A GLOBAL SUPPLY AND DEMAND
PLAN.
SO WITH THAT, I'D LIKE TO THANK
YOU FOR YOUR ATTENTION AND OPEN
UP FOR QUESTIONS.
>> THANK YOU SO MUCH, SIMON, FOR
THIS WONDERFUL TALK.
I SEE ONE QUESTION IN THE Q & A
BOX SO I'LL DIRECT THAT TO YOU.
SO THE QUESTION SAYS, THE FDA
HAS THE ANIMAL RULE FOR
APPROVING DRUGS BASED ON ANIMAL
STUDIES.
IS THERE ANY MOVEMENT ON THE
PART OF THE FDA OR OTHER
REGULATORY AGENCIES AROUND THE
WORLD TO THE ESTABLISHMENT OF A
"CHIP RULE"?
>> YES, I THINK IN ONE OF MY
SLIDES, I ACTUALLY HAD A SCREEN
SHOT OF A BILL THAT'S PASSING
THROUGH AT THE MOMENT IN THE
U.S. IN TERMS OF ACCEPTANCE TO
MICROPHYSIOLOGICAL SYSTEM DATA.
I THINK IT'S IMPORTANT WHEN WE
TALK ABOUT THE ANIMAL RULE THAT
IF THERE'S ONGOING INFECTION,
THEN THE ANIMAL RULE CANNOT
APPLY BECAUSE YOU CAN'T ACTUALLY
GO AND DO A CLINICAL TRIAL.
BUT I THINK WE'RE AT A MOMENT IN
TIME WHERE MICROPHYSIOLOGICAL
SYSTEMS ARE BEGINNING TO BE
ACCEPTED AND THAT'S WHY I'M
HERE.
>> THANK YOU, SIMON.
THERE'S MULTIPLE OTHER QUESTIONS
COMING IN.
SO WE'LL TAKE ONE FROM MARK,
WHO'S ASKING, CAN YOU COMMENT ON
THE VALUE OR NOT OF ANIMAL --
MPS TO SUPPORT THE DEVELOPMENT
OF ANTIVIRAL DRUGS?
>> YES, I THINK THERE ARE
EXISTING DATASETS THAT HAVE BEEN
GENERATED IN NON-HUMAN PRIMATES.
HISTORICALLY.
AND WE HAVE DATASETS THAT HAVE
BEEN PRODUCED IN HUMAN
MICROPHYSIOLOGICAL SYSTEMS, AND
IT MAY BE IMPORTANT TO LINK
HISTORIC DATASETS THAT HAVE BEEN
GENERATED IN NHPs IN HUMANS --
MICROPHYSIOLOGICAL SYSTEM DATA
IN HUMANS, MICROPHYSIOLOGICAL
SYSTEM DATA IN ANIMALS AND THE
ANIMAL DATA AND HUMAN DATA AND
IT MAY BE THAT WE CAN LINK ALL
THOSE FOUR DAY IT SETS.
DATASETS.
SO YES, THERE IS VALUE IN
DEVELOPING THOSE IN ORDER TO
ACCESS THE EXISTING DATASETS
THAT ARE IN ANIMALS AND HUMANS.
>> THANK YOU, SIMON.
NEXT QUESTION IS, IS IT ONLY
CHIP-BASED MODELS THAT WILL BE
CONSIDERED OR CO-CULTURED
WITHOUT A CHIP?
>> I THINK IN SOME OF THE
PRELIMINARY TALKS, WE HEARD
ABOUT SCALE, AND CAN YOU HAVE
THE MOST SOPHISTICATED -- YOU
CAN HAVE THE MOST SOPHISTICATED
SYSTEM ON THE PLANET BUT WITHOUT
SCALE, WE CAN'T ACTUALLY APPLY
IT.
WE NEED THE ACTUAL NUMBERS TO
GET STATISTICAL SIGNIFICANCE.
SO I WOULDN'T ELIMINATE ANY
APPROACH, I THINK, AS LONG AS
IT'S SCIENTIFICALLY JUSTIFIABLE
AND ADDRESSES THE REQUIREMENTS
OF THE DRUG WHICH MAY VARY FROM
ONE DRUG TO ANOTHER, THEN I
DON'T THINK THAT'S IMPORTANT.
I THINK IT'S MORE IMPORTANT TO
MAKE SURE WE HAVE A MODEL THAT'S
REPRODUCIBLE AND RELIABLE AND
CAN COPE WITH THE NUMBERS THAT
ARE REQUIRED TO ACHIEVE
STATISTICAL SIGNIFICANCE.
SO I'M NOT ELIMINATING ANY
APPROACH.
>> MAKE SENSE.
THANK YOU, SIMON.
WE HAVE MORE QUESTIONS COMING
IN.
THE NEXT QUESTION IS, WHAT IS
THE IMPORTANCE OF PROVIDING A
TRUE BLOOD AND LYMPHATIC CIRCUIT
USING WHOLE BLOOD TO PERFUSE THE
MPS SYSTEM?
>> I'VE GOT TO SAY, THAT SOUNDS
ATTRACTIVE.
BUT I KNOW FROM PREVIOUS
EXPERIENCE FROM OTHER STUDIES
THAT THAT CAN BE COMPLICATED
BECAUSE IF, FOR EXAMPLE, SOMEONE
IS RECEIVING ANTIVIRAL DRUG AND
YOU TAKE A SAMPLE OF THE WHOLE
BLOOD, THEN THEY WILL HAVE THAT
ANTIVIRAL DRUG IN THEIR BLOOD
EQUALLY IF THEY'VE HAD -- IF
THEY HAVE AN UNDERLYING
CONDITION SUCH AS A COMPLEMENT
DEFICIENCY, THEN THAT MODEL WILL
REFLECT THAT.
AND SO THE ANSWER TO THAT
QUESTION IS, WHAT IS THE
QUESTION THAT YOU'RE ANSWERING?
AND IF IT'S IS THE INDIVIDUAL
PROTECTED, THEN MAYBE A WHOLE
BLOOD SAMPLE WOULD GIVE YOU THAT
ANSWER.
BUT IF YOU WANT TO TEST WHETHER
A DRUG HAS AN IMPACT ON A
HEALTHY IMMUNE SYSTEM, YOU MAY
HAVE TO HAVE A HEALTHY IMMUNE
SYSTEM WHICH YOU CAN APPLY THAT
DRUG OR THERAPEUTIC TO.
SO I THINK IN ANSWER TO THAT
QUESTION, I THINK IT SOUNDS
ATTRACTIVE, BUT I'M ALSO AWARE
THAT RED BLOOD CELLS REPRESENT
QUITE A TECHNICAL CHALLENGE IN
SOME SYSTEMS, BUT THAT CHALLENGE
CAN BE OVERCOME.
>> UNDERSTOOD.
THANK YOU, SIMON.
THREE MORE QUESTIONS AND WE HAVM
LIVE IF THAT'S OKAY WITH YOU.
>> THAT'S FINE.
WITH BUT PLEASE READ THEM BACKI
CAN'T SEE THEM.
>> ALL RIGHT.
SO THE NEXT QUESTION IS, WHAT
STRATEGIES DO YOU PROPOSE FOR
INCORPORATING ADAPTIVE IMMUNITY?
>> OKAY.
THAT'S A VERY CHALLENGING
QUESTION.
BUT I'M AWARE THAT THERE ARE
SYSTEMS OUT THERE THAT CAN USE
VERY SMALL VOLUMES OF BLOOD TO
ASSESS T-CELLS.
AND I THINK THAT USING MICRO
FLUIDICS, THERE MUST BE A WAY TO
GET A SMALL VOLUME OF WHOLE
BLOOD INTO THE ENDOTHELIAL
CHANNEL OF A SYSTEM WHICH HAS AN
ENDOTHELIAL AND MUCOSAL SURFACE.
I'M NOT A MICRO FLUIDICS EXPERT
BUT I CAN SEE PEOPLE ARE DOING
THAT FOR OTHER PURPOSES, SO I
THINK WHERE THERE'S A NEED,
THERE WILL BE A SOLUTION.
AND SO I DON'T HAVE A GENERIC
ANSWER TO THAT, BUT I DO THINK
THAT SMALL VOLUMES CAN BE
INTRODUCED EITHER INTO THE
ENDOTHELIAL CHANNEL OR IF IT'S
IN AN ORGAN, IT MAY BE
INTRODUCING CELLS.
BECAUSE ACTUALLY WHEN YOU THINK
ABOUT A MICRO NEUTRALIZATION
ASSAY, ALL THAT'S HAPPENING IS
THE ANTIBODIES BEING MIXED WITH
THE VIRUS ARE BEING -- ON TO THE
VERO CELL.
THAT'S PRETTY SIMPLE.
BUT IF WE CAN GET A COMPLEX
TISSUE MODEL AT A CONTROLLED
RATE, MAYBE FLOW CYTOMETRY CAN
GIVE US THE NUMBERS SO WE CAN
STANDARDIZE THE NUMBERS GOING
IN, AND MAYBE THAT CAN BE A WAY
TO ACTUALLY ASSESS ANTIBODIES
AND CELLULAR IMMUNITY AT THE
SAME TIME.
>> THANK YOU, SIMON.
AND HOPEFULLY WE CAN ANSWER SOME
OF THESE QUESTIONS BETWEEN TODAY
AND TOMORROW WITH ALL THE
SPEAKERS, THE TALKS, THE
FANTASTIC TALKS THAT WE HAVE.
ALL RIGHT.
MORE QUESTIONS FOR YOU, SIMON.
SO I'VE GOT ONE THAT SAYS, HI,
SIMON.
THANK YOU FOR THE VERY NICE
TALK.
CAN YOU EXPAND ON WHAT MODEL
COMPONENTS ARE NECESSARY FOR
MICROPHYSIOLOGICAL SYSTEMS BEING
USED FOR VACCINE DEVELOPMENT?
>> AT THE MOMENT, LET ME TALK
ABOUT SARS-COV-2 2, THERE IS A
HEAVY RELIANCE ON HUMAN
IMMUNITY.
BECAUSE THAT'S SEEN AS A GOOD
CORRELATE, A CO-CORRELATE OF
PROTECTION, BECAUSE IT WOULD BE
VERY ODD FOR A VACCINE TO INDUCE
HUMAN IMMUNITY WITHOUT CELLULAR
IMMUNITY.
AND PART OF THAT IS BECAUSE IT'S
ACTUALLY VERY DIFFICULT TO TAKE
THE SAMPLES THAT ARE REQUIRED TO
ACTUALLY CONDUCT CELLULAR
IMMUNITY ASSESSMENT, BUT JUST
BECAUSE IT'S DIFFICULT DOESN'T
MEAN WE CAN'T DO IT.
I THINK THERE IS A -- FOR
CELLULAR IMMUNITY TO BE ANALYZED
BUT FIRST YOU'VE GOT TO THINK
ABOUT HOW DO YOU TAKE A BLOOD
SAMPLE FROM SOMEONE IN AN
ENDEMIC COUNTRY, AND ANALYZE IT?
YOU CAN EITHER SHIP IT BACK TO A
HUB IN A CENTRAL REGION, OR YOU
CAN TAKE THE TECHNOLOGY TO THE
ENDEMIC SITE.
THE SECOND OPTION IS OBVIOUSLY
THE OPTION WHICH WHO WOULD
PROMOTE, THEY WANT TO INCREASE
THE ABILITY OF ENDEMIC COUNTRIES
TO ANALYZE THEIR OWN SAMPLES.
WE HAVE FACED CHALLENGES NOW IN
TERMS OF -- PROTOCOL IN SHIPPING
SAMPLES OUT OF THIRD WORLD
COUNTRIES INTO FIRST WORLD
COUNTRIES.
SO WHAT WE NEED TO DO IS PIONEER
THE TECHNOLOGY OF HOW TO DO THIS
AT HOME, THEN LEARN HOW TO
MINIATURIZE IT AND MAKE IT
FIELD-ACCESSIBLE.
AND THAT'S A SEQUENTIAL PROCESS
AND IT'S NOT GOING TO HAPPEN
OVERNIGHT.
SO THE ANSWER IS, I DON'T HAVE
THE ANSWER YET, BUT I THINK THAT
THERE'S MOMENTUM CERTAINLY IN
THE TALKS I SEE LINED UP IN THE
NEXT TWO DAYS.
MAYBE SOME OF THE ANSWERS YOU'RE
SEARCHING FOR WILL ACTUALLY BE
ANSWERED IN THE COURSE OF THE
PRESENTATIONS, AND IF THEY'RE
NOT, IT'S SOMETHING I'M GOING TO
BE LOOKING AT IN THE VERY NEAR
FUTURE.
>> FANTASTIC.
THANK YOU, SIMON.
SO THIS IS AMY FROM NIAID AGAIN.
THIS IS THE PERSON THAT ASKED
ABOUT THE STRATEGIES THAT YOU'RE
PROPOSING FOR INCORPORATING
ADAPTIVE IMMUNITY, AND SHE'S
EXPANDING AND SAYING, I'M
CONCERNED ABOUT THE BARRIER OF
NHC MATCHING.
>> YES, I KNOW, WE'VE TALKED
ABOUT THAT, AND WHEN WE TALK
ABOUT NHC MATCHING, I THINK IF
YOU TALK ABOUT DELAYED TYPE
HYPERSENSITIVITY, THAT TAKES
SOME TIME TO KICK IN, AND
CERTAINLY WITH REJECTION, IT
DOESN'T HAPPEN IN THE COURSE OF
SEVERAL HOURS -- TAKES QUITE A
BIT OF TIME AND SOME OF THESE
ASSAYS COULD TAKE AN HOUR TO BE
CONDUCTED.
SO ALTHOUGH THEY WOULD OBVIOUSLY
BE SOMETHING LIKE THAT BEING
INITIATED IN THE CASE OF A
MISMATCH.
I THINK THAT SOME OF THE ASSAYS
THAT WE'RE LOOKING AT WOULD
BE -- TO BE CONDUCTED WITHIN A
FEW HOURS OR EVEN OVERNIGHT, IN
WHICH CASE THE --
HYPERSENSITIVITY WILL NOT HAVE
HAD TIME TO INTERFERE WITH THE
ASSAY, BUT IT'S SOMETHING WE ARE
ALL THINKING ABOUT.
>> THANK YOU, SIMON.
TWO MORE QUESTIONS.
ONE MORE SAYS TO DATE, WHAT IS
THE MOST COMPLEX MPS IN TERMS OF
ITS DIVERSITY OF IMMUNE CELLS
INCORPORATION, AND WHAT IMMUNE
CELL TYPES ARE USED FOR YOU --
THINK SHOULD BE INITIALLY
COMBINED FOR TESTING?
>> I THINK I CAN'T ANSWER THAT
QUESTION, I THINK SOME PEOPLE
HAVE ACTUALLY ANSWERED THAT
QUESTION, SO I THINK I'D HAVE
THE RIGHT TO DEFER THAT
QUESTION.
I KNOW NATURAL KILLER KELCE ARE
OBVIOUSLY VERY IMPORTANT, BUT --
AND THERE ARE MANY T-CELLS THAT
MAY BE INVOLVED AND
UNFORTUNATELY WE DON'T REALLY
KNOW WHICH ONES ARE YET.
WE'RE GETTING HINTS FROM IB IN
VITRO ANALYSIS FLOW CYTOMETRY.
BUT UNTIL WE HAVE MODELS TO
ANALYZE IT, WE WON'T HAVE THE
ANSWERS TO THAT.
>> THANK YOU, SIGH SIMON.
I INVITE ANYONE ON THE PANEL
TODAY TO TYPE ANSWERS AS WELL.
I KNOW THERE'S MULTIPLE PEOPLE
ON THE PANEL RIGHT NOW THAT
MIGHT HAVE ADDITIONS TO THE
ANSWER AS WELL.
SO FEEL FREE TO TYPE IN YOUR
ANSWERS.
ONE MORE QUESTION.
A PAPER IN THE JOURNAL OF
INFECTIOUS DISEASES EXPLORED
NOROVIRUS INFECTION MECHANISMS
BY USING STEM CELL DERIVED
INTESTINAL -- TO SIMULATE
INFECTION.
HOWEVER, IT WAS FOUND ONLY
CERTAIN NOROVIRUS GENOTYPES WERE
ABLE TO INTBECT THE --.
ARE YOU FAMILIAR WITH OTHER
SCENARIOS IN WHICH STEM CELL
DERIVED MODEL SYSTEMS
ENCOUNTERED SIMILAR LIMITATIONS?
>> I'M NOT FAMILIAR WITH OTHER
SCENARIOS THAT HAVE FAILED, BUT
I AM AWARE THAT IF WE KEEP USING
VOLUNTEERS TO SUPPLY OUR SEEDS
CULTURES FOR MICROPHYSIOLOGICAL
SYSTEMS, WE'RE GOING TO GET A
LOT OF VARIANTS, AND IT MAY BE
THAT FOR SEARCH IN ORGANISMS, WE
NEED TO MAKE SURE THE
APPROPRIATE BIOMARKERS ARE
REPRESENTED IN THE MODEL THAT WE
USE.
IT MAY BE THAT -- X CELL Y IS
APPROPRIATE BUT WE'RE GOING TO
HAVE TO CHANGE THE CELLS.
BUT WE NEED TO ENSURE THAT THE
MODEL IS SUITABLE FOR THE
QUESTION THAT WE'RE ASKING.
FOR YEARS NOW, WE'VE BEEN USING
VERO CELLS.
AND I DON'T THINK THEY'RE A VERY
GOOD MODEL OF SARS-COV-2
INFECTION, AS YOU MAY HAVE
GATHERED.
BUT WHETHER OR NOT VERO CELLS
ARE APPROPRIATE FOR VIRUS X WILL
DEPEND ON THE RESULT THAT WE
ACHIEVE AND WHAT BIOMARKERS ARE
REQUIRED FOR VIRUS X.
BUT I THINK IN THE SAME WAY THAT
YOU WOULDN'T USE YOUR STANDARD
SCIENTIFIC TECHNIQUE TO APPLY TO
ANY VIRUS, YOU'D WANT TO MAKE
SURE THAT THE SCIENTIFIC
TECHNIQUES OR SERVICES YOU HAVE
ARE APPROPRIATE FOR USE.
AND THAT'S PART OF THE
VALIDATION OF THE MODEL.
AND IT MAY BE THAT WE NEED TO
VALIDATE A GENERIC MODEL FOR
SUITABILITY TO THE LIST OF
PRIORITY PATHOGENS THAT WAS
LISTED EARLIER ON BY EMILY.
AND IT MAY BE THAT FOR THE
PRIORITY PATHOGENS, MODEL X WILL
BE APPROPRIATE FOR THOSE BUT NOT
FOR MODEL -- NOT FOR VIRUS Y,
AND IT MAY BE THAT SPECIFIC
VIRUSES HAVE SPECIFIC BIOMARKER
REQUIREMENTS THAT HAVE TO BE
PRINT.
AND IT MAY BE THAT
PRESENT.
IT MAY BE THE FIRST TIME YOU TRY
IT WHEN YOU GROW IN THE CELLS
FOR A COUPLE OF WEEKS, MAYBE YOU
HAVEN'T GOT THE RIGHT BIOMARKERS
BUT MAYBE YOU'RE MISSING A VITAL
COMPONENT IN THE DIFFERENTIATION
MEDIA THAT SIMPLY NEEDS
MODIFICATION AND IT MAY BE
THROUGH A QUALITY MEASUREMENT
SYSTEM, THOSE FAILURES ARE
EXTREMELY IMPORTANT TO PUBLISH,
BECAUSE IT MAY BE THAT WE SAY,
WELL, WE NEED TO GO BACK AND SEE
WHY THAT'S FAIL AND SHARE THAT
LEARNING WITH THE COMMUNITY SO
THAT WE ALL KNOW THAT THE
BIOMARKER REQUIREMENT FOR THE
NOROVIRUS HAS TO BE INDUCED BY
THIS MEDIA -- WE CAN IMMEDIATELY
FIND THAT OUT OURSELVES AS A
COMMUNITY AND SHARE THAT.
>> FANTASTIC.
>> IT'S THROUGH FAILURE WE CAN
LEARN, SO FAILURE IS PART OF THE
NATURAL PROCESS OF IMPROVEMENT.
>> THANK YOU, SIMON.
WE WILL HAVE A TALK TOMORROW ON
NOROVIRUS SO WE MAY GET MORE
ANSWERS FROM FOLKS THAT ARE
INTERESTED TO LEARN MORE.
OKAY.
I'M GOING TO STOP HERE.
YOU DO HAVE MORE QUESTIONS,
SIMON.
SO FEEL FREE TO, AFTER WE --
DURING THE LUNCH BREAK, IF YOU
CAN STAY A LITTLE LONGER AND
MAYBE ANSWER SOME OF THESE
QUESTIONS AND TYPE IN THE
ANSWERS.
ALSO I INVITE EVERYONE ON THE
PANEL IF THEY WOULD LIKE TO ADD
ANYTHING TO THE ANSWERS, FEEL
FREE TO TYPE IN YOUR ANSWERS AS
WELL.
AND THEN PLEASE, THE AUDIENCE,
KEEP ASKING QUESTIONS.
THIS IS GREAT.
WE'D LOVE TO GET YOUR QUESTIONS,
WE'D LIKE TO HAVE NICE
DISCUSSIONS DURING THIS MEETING.
OKAY.
SIMON, THANK YOU SO MUCH FOR
SUCH A WONDERFUL PRESENTATION,
SO ENGAGING, SO MANY QUESTIONS.
WE'RE REALLY HAPPY YOU WERE
HERE, AND WE APPRECIATE YOUR
TIME DURING THIS.
ALL RIGHT.
SO I'M GOING TO -- WE'RE GOING
TO GO FOR A LUNCH BREAK NOW, AND
WE ARE GOING TO RECONVENE AT
1:20 FOR SESSION 1 ON 3D TISSUE
MODELS, UTILITY AND LIMITATIONS.
HOPEFULLY YOU'RE ENJOYING TODAY
AND YOU ARE GOING TO ENJOY
SESSION 1.
SO HAVE A GREAT LUNCH BREAK, AND
LET'S RECONVENE AT 1:20.
I'M GOING TO PULL UP THE AGENDA
HERE SO PEOPLE ARE AWARE.
SO 1:20 P.M., SESSION 1 ON 3D
TISSUE MODELS: UTILITY AND
LIMITATIONS.
THANK YOU.
>>WELCOME BACK, EVERYONE.
NEXT WE ARE GOING TO HAVE
SESSION 1 WHICH IS ON 3D TISSUE
MODELS AND THEIR UTILITY AND
LIMITATIONS.
THIS SESSION IS CHAIRED BY
DR. MARK FERRER, CURRENTLY THE
DIRECTOR OF THE 3D TISSUE
BIOPRINTING LABORATORY AT THE
NATIONAL CENTER OF ADVANCING
TRANSLATIONAL SCIENCES AT NIH,
WHICH IS A MULTIDISCIPLINARY
GROUP ESTABLISHED IN 2018 WITH
THE GOAL OF CREATING, VALIDATING
AND USING 3D BIOENGINEERED
TISSUES FOR DISEASE MODELING AND
PREDICTIVE DRUG DISCOVERY AND
DEVELOPMENT.
THANK YOU FOR BEING HERE, MARK,
AND I'LL PASS THE BATON TO YOU.
>> THANKS SO MUCH, IS A SARI.
WELCOME, EVERYONE, BACK TO
SESSION 1.
I WANT TO THANK THE SPEAKERS FOR
MAKING THE TIME TO TELL US ABOUT
THEIR RESEARCH, ESPECIALLY
CHRISTINE AND HANS FROM EUROPE,
WE KNOW IT'S LATE OVER THERE.
AS SARINE MENTIONED AT THE
BEGINNING, THE OBJECTIVE OF THIS
SESSION WAS TO REALLY GIVE AN
OVERVIEW NOT SPECIFICALLY FOR
VIRUSES BUT IN GENERAL OF WHAT
TISSUE MODELS ARE AVAILABLE AND
DISCUSS THE CHALLENGES IN
BUILDING THESE MODELS AS WELL AS
IMPLEMENTING THEIR USE FOR DRUG
SCREENING AND DEVELOPMENT AND
REALLY THE GOOD AND THE BAD.
SO HOPEFULLY WE'LL HAVE TIME TO
GO THROUGH THE TALKS, REMINDER
BECAUSE IT'S 20 MINUTES, SO WHAT
I'LL DO IS I'M GOING TO TURN MY
CAMERA OFF AND THEN TWO MINUTES
BEFORE THE 20 MINUTES, I'LL TURN
IT ON SO THAT WILL BE THE SIGN
THAT THE -- THE SIGNAL THAT YOU
HAVE 2 MINUTES LEFT.
AND THEN AGAIN, AS SARINE
MENTIONED, WE'RE GOING TO GO
FROM ONE SPEAKER TO THE OTHER
AND THEM WE'RE GOING TO HAVE
HALF AN HOUR AT THE END FOR
QUESTIONS AND DISCUSSIONS.
IF YOU HAVE ANY QUESTIONS ALONG
THE WAY, YOU CAN PUT THEM IN
QUESTIONS AND ANSWERS BOX, AND
THE SPEAKERS ARE WELCOME TO
PROVIDE ANSWERS IN WRITING OR WE
CAN ADDRESS THEM AT THE END AS
WELL.
SO WITHOUT FURTHER ADO, IT IS MY
PLEASURE TO INTRODUCE THE FIRST
SPEAKER OF THIS SESSION,
DR. CHRISTINE MUMMERY, PROFESSOR
OF DEVELOPMENTAL BIOLOGY IN THE
DEPARTMENT OF ANATOMY AT LEIDEN
UNIVERSITY MEDICAL CENTER, WHERE
SHE ALSO HEADS THE FACILITY.
SHE'S A MEMBER OF THE ROYAL
ACADEMY OF SCIENCE AND PAST
PRESIDENT OF SOCIETY OF STEM
CELL RESEARCH.
SHE'S GOING TO TELL US ABOUT HER
RESEARCH IN CARDIOVASCULAR
DISEASES.
CHRISTINE, THE FLOOR IS YOURS.
>> THANK YOU.
CAN YOU SEE MY SCREEN?
>> YEAH, YOU HAVE TO MAKE IT
PRESENTATION MODE BUT OTHERWISE
WE SEE IT.
>> PERFECT.
>> OKAY, THANK YOU.
SO I WAS ASKED TO TALK ABOUT
CARDIOVASCULAR DISEASE MODEL,
AND TOWARDS STANDARDIZATION OF
HOW WE MIGHT USE THEM.
AND IN THE CONTEXT OF VIRUS,
IT'S QUITE IMPORTANT TO DISCUSS
THE LIMITATIONS.
SO I'M GOING TO DISCUSS THAT A
LITTLE BIT, AND HERE IS A SORT
OF OVERVIEW OF WHAT WE CAN DO
WITH PLURIPOTENT STEM CELLS.
SO WE CAN MAKE THEM FORM ALL
DERIVATIVES OF THE BODY, AND IN
MANY CASES, WE CAN DO THAT NOW
IN VITRO VERY SUCCESSFULLY, BUT
WE ALL KNOW THE LIMITATIONS OF
THIS MODEL ARE THAT THE CELLS
ARE ALL IMMATURE.
AND THIS IN THE CONTEXT OF,
LET'S SAY, COVID RESEARCH HAS
NOT BEEN PARTICULARLY BENEFICIAL
BECAUSE IF I WOULD TAKE THE
HEART AS AN EXAMPLE,
CARDIOMYOCYTES FROM IPS CELLS,
FOR EXAMPLE, EXPRESS ACE
2 RECEPTORS, BUT THEY'RE NOT
REALLY EXPRESSED VERY HIGHLY IN
THE ADULT HEART.
SO WE HAVE TO BE VERY CAREFUL IN
INTERPRETING SOME OF THIS DATA.
SO LET ME TELL YOU SOMETHING
ABOUT HOW WE'VE BEEN OVERCOMING
THIS LIMITATION AND ARE TRYING
TO MAKE MATURE CARDIOMYOCYTES
THAT WE CAN MOVE FORWARD IN
MODELING DISEASE, AND I'LL GIVE
YOU SOME EXAMPLES.
SO WHAT DO I MEAN ABOUT IMMATURE
CELLS?
THIS IS BEATING CELLS -- THE
BEATING MEANS THEY ARE IMMATURE
BECAUSE AN ADULT HEART CELL
WOULD NOT BEAT BY ITSELF, IT
WOULD NEED A PACEMAKER.
HERE ARE NO PACEMAKERS SO WE
KNOW IT'S IMMATURE.
IT LACKS A 3D ENVIRONMENT, AND
IT LACKS OTHER CARDIAC-SPECIFIC
CELL TYPES.
SO WE ASKED THE QUESTION, WOULD
IT BE POSSIBLE TO ADD THESE
CELLS IN IN A 3D ENVIRONMENT AND
CREATE SOMETHING THAT LOOKED
MORE LIKE HEART TISSUE.
AND JUST TO BRIEFLY SHOW YOU THE
HEART, THIS IS THE MYOCARDIUM
HERE, LINED WITH AN ENDO CARD
YUM.
THERE'S AN OUTER LAYER OF EP
CARD YUM THAT EP --
FORM ABOUT 70% OF THE CELLS IN
THE HEART.
EVERY CARDIOMYOCYTE TOUCHES A
BLOOD VESSEL SO IT TOUCHES AN
ENDOTHELIAL CELL.
SO WE THOUGHT WE MIGHT MAKE A
MODEL IN WHICH WE INCLUDED THE
ENDOTHELIAL CELLS, CARDIAC
FIBROBLASTS, SO REMEMBER EVERY
TISSUE HAS ITS OWN TYPE OF
STROMA CELLS, IT'S OWN TYPE OF
END THEEL YELL CELLS, ITS OWN
TYPE OF FIBROBLASTS THAT WE
WOULD DERIVE FROM EPICARDIAL
CELLS AND THE CARDIOMYOCYTES.
THE FIRST THING WE NEEDED TO DO
WAS TO ACTUALLY GET THE
DIFFERENTIATION PROTOCOLS VERY
ROBUST, SO WE WANTED TO USE ONE
BASAL MEDIUM TO DERIVE THESE
CELL TYPES AND WHERE POSSIBLE
MOVE AWAY FROM GROWTH FACTORS.
THIS WOULD REDUCE LOT TO LOT
VARIABILITY, WOULD ENABLE
UPSCALING AND REDUCE COSTS.
ESSENTIALLY THAT'S WHAT WE'VE
WORKED ON FOR THE LAST SEVERAL
YEARS, WE HAVE MANY OF THESE
PROTOCOLS HIGHLY DEFINED, SOME
BASED ON SMALL MOLECULES.
WE CAN NOW DERIVE ENDOTHELIAL
CELLS, CARDIOMYOCYTES AND
CARDIOFIBROBLASTS FROM A COMMON
PROGENITOR THAT WE GET FROM IPS
CELLS.
WE CAN NOW CRYOPRESERVE THESE IN
LARGE BATCHES.
WE CAN CARRY OUT QA AND QC
BEFORE FREEZING, AND THESE ARE
READY TO USE IN ASSAYS AFTER
THAW, SO THIS MAKES IT A GREAT
ADVANTAGE FOR DOING ASSAYS.
SO WHAT DO WE MEAN BY
MATURATION?
HOW WOULD WE RECOGNIZE IF IT
OCCURRED IN OUR CARDIOMYOCYTES?
IT'S VERY SIMPLE, WE WOULD LOOK
AT GENE EXPRESSION, LOOKING AT
WHETHER SUPPLIES
WHETHER SPLICE, STRUCTURE,
CONTRACTION, ELECTROPHYSIOLOGY
AND METABOLISM, THEY WILL ALL
DIFFER FROM WHAT WE WOULD SEE IN
A PLURIPOTENT STEM CELL.
THESE ARE CARRIED OUT TO TRY TO
MATURE CARDIOMYOCYTES, ENGINEER
HEART TISSUES, 3D STRETCHING
SORT OF STRUCTURES, CHANGING THE
CULTURE REAGENTS, USING ECK PO
TICK GENE EXPRESSION, USING
SOMETHING CALLED A BIOWIRE, 3D
PRINTING, ALIGNING THE CELLS.
THEY ALL WORK TO A CERTAIN
EXPECT BUT WE WANTED A VERY
SIMPLE LOW TECH METHOD.
SO WE DECIDED TO MAKE SIMPLE
MICRO TISSUES.
SO WE'RE USING THESE
PREDIFFERENTIATED CELLS, WE CAN
DO THESE ASSAYS ISOGENICLY.
WE PUT THEM TOGETHER AS 5,000
CELLS IN THESE RATIOS, SO 70%
CARDIOMYOCYTES, 15% OF EACH OF
THE OTHER CELLS, AND PUT THEM IN
MULTI-WELL PLATES AND WE GET
THESE BEATING HEART CELLS.
THESE MINI HEARTS, YOU COULD
SAY, IN A METAPHOR.
IT'S VERY LOW TECH.
SO WITHOUT SHOWING YOU ALL THE
DATA, THIS IS WHAT OUR MICRO
TISSUES LOOK LIKE.
WE SEE THE CARDIOFIBROBLASTS,
THE ENDOTHELIAL CELLS ARE LINED
UP IN THESE TWO STRUCTURES.
WE CAN COUPLE THEM INTO ORGAN ON
CHIP MODELS AND CREATE A LUMEN
THAT'S FULLY VASCULARIZED AND
THE CARDIOMYOCYTES ARE MARKED
HERE IN GREEN.
WE SHOW THESE HAVE STRUCTURAL
MATURITY, ELECTRICAL AND
MECHANICAL MATURITY AND
METABOLIC MATURITY.
THE FEATURES ACTUALLY OF AN
EARLY POSTNATAL CARDIOMYOCYTE,
SO THEY'RE NOT FULLY ADULTS AND
CERTAINLY NOT OLD, WITH YOU E
POSTNATAL.
USING JUST 5,000 5,000 CELLSR
TISSUE, WE CAN REDUCE THE COST
TO
IN THE RANGE OF WHAT PHARMA
WOULD LIKE TO HAVE FOR DRUG
ASSAYS.
AND WE CAN DO MULTI-LINEAGE
CARDIAC DISEASE MODELING.
WHAT I MEAN BY THAT IS WE CAN
REPLACE ANY ONE OF THOSE THREE
CELL TYPES, THE GUY WROA G,
ENDOTHELIAL CELLS OR RM -- FROM
CONTAINING A MUTATION.
SO IT ALLOWS US TO IDENTIFY
WHICH CELL IS CONTRIBUTING TO A
PHENOTYPE.
I'LL GIVE YOU ONE QUICK EXAMPLE
HERE OF THESE TRI--LINEAGE
MODELS.
IT'S AN EXAMPLE OF A DISEASE
CALLED ARRHYTHMOGENIC
CARDIOMYOPATHY.
IT'S CAUSED BY MUTATIONS OF THE
DESMOSOMAL PROTEIN PKP26789.
IT'S CHARACTERIZED BY
ARRHYTHMIAS AND FIBRO-FATTY
REPLACEMENT OF THE MIGHT OWE
CARD YUM.
THE CELL OF ORIGIN IS UNKNOWN,
POSTULATED THAT CARDIAC
FIBROBLASTS MAY ALSO PLAY A
ROLE.
SO WHAT WE DID WAS TO REPLACE
THE CARDIAC FIBROBLASTS IN THE
MICRO TISSUES WITH THESE ACM
MUTANT CARDIAC FIBROBLASTS
ISOGENICLY.
SO TO CUT A LONG STORY SHORT,
THIS IS AN EXAMPLE OF WHAT THESE
MICRO TISSUES LOOK LIKE AND
WHAT'S SHOWN HERE IS THE
STAINING FOR THE JUNCTIONS.
YOU CAN SEE THE CONTROL HERE ON
THE LEFT, THERE'S A GREAT DEAL
OF FORMATION, AND YOU SEE IN THE
MUTANT CELLS THE CARDIAC
FIBROBLASTS FORM FEWER GAP
JUNCTIONS AND THIS IS CAUSED BY
THE ABSENCE OF PKP2.
BUT MOST IMPORTANTLY, WHEN WE
START PACING THESE CELLS, SO
YOUR HEART NORMALLY BEATS AT
1 HERTZ 60 TIMES A MINUTE, YOU
CAN SEE THE CONTROL MICRO
TISSUES HERE FOLLOWING THIS
PACING VERY NICELY.
IT'S NOT TOO BAD FOR THE ONES
CONTAINING THE MUTANT
FIBROBLASTS BUT IF WE GO TO
2 HERTZ, YOU SEE THEY'RE HAVING
DIFFICULTY FOLLOWING THE PACING
SIGNALS.
AND WHEN WE GO TO 3 HERTZ, WHICH
IS ABOUT WHEN YOU MIGHT BE
RUNNING A MARATHON OR SOMETHING,
YOU CAN SEE HERE THESE VERY
PROMINENT ARRHYTHMIAS.
SO THIS IS TELLING US THAT
ACTUALLY THE CARDIAC FIBROBLASTS
ARE ONE OF THE CULPRITS IN THIS
DISEASE, SO THE CARDIOMYOCYTES
ARE NORMAL, YET WE'RE GETTING
THESE ARRHYTHMIAS.
ANOTHER QUICK EXAMPLE.
MANY OF YOU KNOW THAT KNOWT
DOXORUBICIN IS USED FOR AS YOE
CAR SOMA IN CHILDREN AND BREAST
CANCER.
THE PROBLEM IS MANY PATIENTS
DEVELOP IRREVERSIBLE HEART
FAILURE 2 TO 10 YEARS AFTER
TREATMENT AND THE ONLY THING
THAT CAN BE DONE FOR THESE
PATIENTS IS HEART REPLACEMENT.
SO WHAT WE CAN SEE HERE ON THE
TOP IS VECTORROLOGY SHOWING THE
CONTRACTION OF A SINGLE MICRO
TISSUE HERE, YOU CAN SEE IN THE
GRAPH ON THE RIGHT, THE
CONTRACTION FOLLOWED BY THE
RELAXATION, QUANTIFIED IN THIS
PLOT HERE.
AND AT THE BOTTOM.
YOU CAN SEE WHAT DOXORUBICIN
DOES.
IT REDUCES THE AMOUNT OF
CONTRACTION SIG SIGNIFICANTLY.
NOW A COLLEAGUE OF MINE IN
LEIDEN IS A BRILLIANT MEDICINAL
CHEMIST AND HE FIGURED OUT THAT
DOXORUBICIN ACTUALLY HAD TWO
PARTICULAR BIOLOGICAL EFFECTS.
IT CAN CAUSE DNA DAMAGE, AND IT
CAUSES HISTONE EVICTION.
AND IT'S THE DNA DAMAGE THAT
ACTUALLY CAUSES THE DAMAGE TO
THE HEART AND THE HISTONE
EVICTION IS THE ACTIVITY
RESPONSIBLE FOR KILLING THE
TUMOR.
SO HE DESIGNED VARIANTS OF
DOXORUBICIN WHICH ONLY HAVE THE
HISTONE EVICTION PART AND NOT
THE DNA DAMAGE.
AND THIS KILLED THE TUMORS, AND
WHAT WE SAW VERY NICELY IN THIS
MODEL OF THE MICRO TISSUES, HERE
YOU CAN SEE THE AMPLITUDE OF
CONTRACTION OF THE CONTROLS,
HERE YOU SEE WITH DOXORUBICIN,
BUT WHEN WE ADD THIS VARIANT,
ACLARUBICIN, THERE'S NO EFFECT
ON THE CONTRACTION.
WE'RE DISCUSSING WITH THE
AUTHORITIES IF THEY WOULD FIND
THIS ACCEPTABLE AS AN OPTION
INSTEAD OF A SECOND ANIMAL MODEL
TO THE MASS.
THEY DON'T ACTUALLY FIND IT
ACCEPTABLE, BUT THEY FOUND IT
VERY INTERESTING TO LOOK AT.
A THIRD EXAMPLE IS HOW WE CAN
NOW USE THIS FOR HIGH-THROUGHPUT
SCREENING.
AS I SAID, THESE TISSUES ARE
VERY EASY TO MAKE, WE CAN DO IT
WITH ROBOTICS NOW, PUT THEM IN
THESE LARGE WELL PLATES, A WE
CAN LOOK AT CALCIUM SIGNALING.
SO YOU CAN SEE THAT HERE, WE
LOADED THEM WITH A CALCIUM DYE,
YOU CAN SEE THEM FLASHING AND
YOU CAN SEE IN A HIGH-THROUGHPUT
SYSTEM WHERE THE DRUGS MIGHT
HAVE THIS EFFECT.
SO THIS IS WHAT WE WOULD SEE IF
THERE'S ARRHYTHMIAS AND THIS IS
WHAT WE WOULD SEE IF IT WAS
OKAY.
SO LOOKING AT YET ANOTHER
DISEASE, THIS IS CALLED CPVT.
IT ALSO CAUSES ARRHYTHMIAS BUT
IT'S CAUSED BY MUTATIONS IN A
RECEPTOR THAT REGULATES CALCIUM
HANDLING.
YOU CAN SEE HERE IN THAT SYSTEM
I JUST SHOWED YOU, THAT THERE'S
REGULAR CALCIUM BEATS.
IF WE GIVE THE CPV T-CELLS A
TRIGGER, YOU CAN SEE THESE
ARRHYTHMIAS REVEALED.
AND IF WE ADD A DRUG THAT WORKS
IN SOME PATIENTS BUT NOT OTHERS,
WE CAN SEE WE GET THIS RESCUE.
NOW IT'S BEEN VERY HARD IN ANY
OTHER MODELS TO ACTUALLY SEE THE
EFFECTS OF FLECANYDE.
WE HOPE NOW AND IN THE FUTURE WE
CAN DO POPULATION STUDIES AND
FIGURE OUT WHICH PATIENT IS
LAKELY TO RESPOND AND DO SCREENS
OF LIBRARIES TO SEE WHICH DRUGS
MIGHT BE EFFECTIVE.
MOVING NOW TO VASCULAR DISEASE,
VASCULAR DISEASE IS OFTEN CAUSED
BY A FAILURE TO HAVE PROPER
DIALOGUE BETWEEN THE CELLS THAT
MAKE UP THE VESSEL WALL.
SO THAT'S THE VEST COLLAR SMOOTH
MUSCLE CELLS, ENDOTHELIAL CELLS
AND PERHAPS THE IMMUNE SYSTEM IN
THE STROMA.
SO MUTATIONS IN ANY OF THESE
SIGNALING PATHWAYS BETWEEN THESE
CELLS CAUSE A VARIETY OF RARE
VASCULAR DISEASES.
TOGETHER, THERE ARE ACTUALLY
QUITE A LOT OF PEOPLE WITH THESE
CONDITIONS.
WE'RE PARTICULARLY INTERESTED IN
ONE CALLED HEREDITARY
HEMORRHAGIC TELANGIECTASIA.
THIS IS CAUSED BY DISTRACTION OF
A DIALOGUE BETWEEN THE
ENDOTHELIAL CELLS AND THE
PARASITES.
SO LOOKING TOWARDS
STANDARDIZATION, THIS IS VERY
IMPORTANT, SO WE CAN GET
CD31 POSITIVE ENDOTHELIAL CELLS
FROM IPS CELLS QUITE HAPPILY.
THEY SAY PROPERLY WITH THE RIGHT
MARKER SO HERE IN PECAM1 VON
WILLEBRAND FACTOR, AND WE CAN
STANDARDIZE THESE IN QA AND QC
BY QUANTIFYING FUNCTIONAL
PARAMETERS.
I'LL JUST SHOW YOU AN EXAMPLE OF
WHAT THAT MEANS AND HOW WE CAN
MONITOR BATCH TO BATCH
VARIABILITY.
SO WE USE SOMETHING CALLED
ELECTRICAL CELL-SUBSTRATE IMPEE
DANCE SENSING, ECIS, AND IT
MEASURES THE BARRIER FUNCTION OF
THE EPITHELIAL LAYER.
SO BASICALLY WHAT YOU DO IS YOU
MEASURE THE RESISTANCE, YOU
CREATE A WOUND IN THAT
EPITHELIAL LAYER BY TURNING UP
THE GAIN ON SOME ELECTRODES, IT
BYRNES A HOLE YOU AND GET
MIGRATION OF THE CELLS BACK INTO
THAT WOUND.
YOU DO THIS IN COMPLETE MEDIUM,
SERUM-FREE MEDIUM OR MEDIUM IN
VEGF.
SO THIS IS THE MIGRATION SHOWN
AND THE IMPEE DANCE.
IMPEDANCE.
NOW JUST WATCH THAT SHAPE.
I'M NOT GOING TO GO INTO THE
DETAILS TOO MUCH, BUT THIS IS
INDEPENDENT BIOLOGICAL
EXPERIMENTS WITH THE SAME BATCH
OF IPS ENDOTHELIAL CELLS.
SO BASICALLY WHAT YOU SEE IS THE
SHAPE IN ALL CASES IS THE SAME,
SO THE RESISTANCE IN ALL THREE
BATCHES IS THE SAME.
SO THAT'S GOOD.
WITHIN ONE PATCH.
SO BAMP.
SO THERE'S NOTHING TECHNICALLY
WRONG WITH THE ASSAY.
IF WE TAKE INDEPENDENT BATCHES
OF THESE CELLS ISOLATED FROM THE
SAME CONTROL LINE, WE CAN'T NO
DIFFERENCES AT ALL.
SO THE BATCH TO BATCH
VARIABILITY IS LOOKING EXTREMELY
GOOD.
WHEN WE GO TO LINE TO LINE
DIFFERENCES, THAT'S WHEN WE'RE
SEEING A DIFFERENCE.
YOU CAN SEE HERE WITH THREE
DIFFERENT LINES, THERE ARE TWO
LOOKING QUITE SIMILAR, ONE
SLIGHTLY DIFFERENT IN THE
RESISTANCE IT'S SHOWING US IN
THE TIER ASSAY, TRANS EPITHELIAL
RESISTANCE ASSAY.
SO THAT MEANS WE HAVE TO BE VERY
SURE TO TAKE THE RIGHT CONTROL
BELONGING TO EACH LINE BEFORE WE
CAN MAKE CONCLUSIONS.
SO HAVING DONE THAT SORT OF QA
AND QC IN THESE VERY LARGE
BATCH, WE CAN PUT THEM IN AN
ORGANOID CHIP FORMAT.
THIS IS USING COMMERCIALLY
AVAILABLE BIOCHIP CHIP.
YOU HAVE A GEL CHANNEL IN THE
MIDDLE OF THESE MICRO FLUIDIC
DEVICE, GEL CHANNEL CONTAINING
THE CELLS.
THERE ARE TWO MEDIA CHANNELS ON
EITHER SIDE.
THEY COME IN MULTIPLEX FORMS.
WE PUT A SUSPENSION OF
ENDOTHELIAL CELLS AND PARASITES
IN THE MIDDLE CHANNEL, AND THESE
ENDOTHELIAL AND SMOOTH MUSCLE
CELLS ORGANIZE THEMSELVES INTO A
VASCULATURE.
IF THIS FILM COULD RUN, YOU
COULD SEE IT.
BUT IT SELF-ORGANIZES HERE, AND
YOU CAN ACTUALLY SEE THESE
FORMING LEUM NIEZED VESSELS VERY
NICELY.
HERE IN PURPLE YOU CAN SEE THE
EPITHELIAL CELLS AND BEAUTIFUL
LUMEN FORMED INSIDE.
THESE ARE THE EPITHELIAL CELLS
IN PURPLE AND THE GREEN CELLS
ARE THE SMOOTH MUSCLE CELLS.
THIS IS IN A CONTROL SITUATION
SO VERY TIGHT CONTACT BETWEEN
THESE SMOOTH MUSCLE CELLS AND
THE ENDOTHELIAL CELLS.
NOW, IN THIS DISEASE CALLED HHT,
WHICH I SAID WAS ONE OF OUR
FAVORITE DISEASES, THERE ARE
MUTATIONS IN THE SIGNALING
PATHWAY.
THESE PATIENTS HAVE VERY WEAK
BLOOD VESSELS, THEY HEMORRHAGE
VERY EASILY, THEY HAVE
PARTICULARLY CHRONIC NOSE
BLEEDS, SO MAYBE FIVE TO 10
TIMES A DAY, SOMETIMES UP TO
HALF A LITER A TIME.
THEY'RE COMPLETELY MISERABLE.
SO WHAT WE'RE LOOKING FOR IS
WAYS TO STABILIZE THESE VESSELS.
THE FIRST QUESTION IS, COULD WE
CREATE A MODEL TO MIMIC THIS?
AND BASICALLY WHAT WE'VE DONE IS
SHOWN IN THE NEXT SLIDE.
WE CAN CREATE THIS MODEL.
SO IN THE TOP, YOU SEE AGAIN
THIS CONTROL, THE PURPLE ON THE
RIGHT IS THE ENDOTHELIAL CELLS,
THE GREEN OF THE SMOOTH MUSCLE
CELLS.
BUT WHEN WE HAVE THE CELLS
FORMED FROM IPS CELLS FROM AN
HHT PATIENT, WHAT WE FIND IS
THESE SMOOTH MUSCLE CELLS
INTERACT VERY POORLY WITH THE
ENDOTHELIAL CELLS, AND THAT
STABILITY IS LIKELY THE REASON
THAT THESE VESSELS HEMORRHAGE
EASILY.
AND WHEN WE LOOK AT THE FLOW OF
DEXTRAN THROUGH THESE LEUM
NIEZED VESSELS ON THE TOP, IT
RETAINED WITHIN THESE VESSEL
STRUCTURES WITH BUT THE HHT-D
VESSELS ARE LEAKING, JUST AS
THEY ARE IN PATIENTS.
WE'RE USING THIS ASSAY TO SCREEN
FOR REPURPOSED DRUGS AT THE
PRESENT TIME AND WE'VE GOT -- ON
THE BASIS OF THIS ASSAY, WE NOW
HAVE TWO DRUGS IN CLINICAL
TRIAL.
SO JUST TO SUMMARIZE BECAUSE
TIME FLIES, THIS IS -- WHAT I'VE
SHOWED YOU IS A LOW-TECH
SCALABLE AND INEXPENSIVE MODEL
TO STUDY CARDIAC DISEASE AND
TOXICITY BASED ON MATURE
CARDIOMYOCYTES, SO I THINK IF WE
MOVE NOW TO LOOKING MORE AT THE
EFFECT OF VIRUSES, WE MAY SEE
THE OUTCOME WE EXPECT.
THE MULTICELL TYPE MODEL ALLOWS
IDENTIFICATION OF CELLULAR
TARGET IN CARDIAC DISEASE.
THE 3D ORGANS ON CHIP UNDER FLOW
THAT I SHOWED YOU CAPTURE SMALL
VESSEL DISEASE MODEL PHENOTYPES.
AND WE DO NOT SEE THIS IN 2D
BECAUSE IN 2D, THERE ARE NO
LUMEN.
AND MANY IPS AND ORGANOID MODELS
ON WHICH THEY'RE BASED ARE READY
TO GO FOR EXAMINING EFFECT OF
VIRUSES, DRUG REPURPOSES AND
SCREENING OF THERAPEUTIC DRUGS.
SO THESE ARE THE PEOPLE IN THE
LAB WHO HAVE BEEN DRIVING THIS
WORK, JUNIOR PIs, THESE ARE
PH.D. STUDENTS AND POSTDOCS
INVOLVED IN THE WORK AND THESE
ARE SOME OF OUR MANY
COLLABORATOR.
THANK YOU.
AND OF COURSE OUR FUNDERS AT THE
BOTTOM.
>> THANK YOU SO MUCH, CHRISTINE.
I LOVE YOUR LOW TECH MODEL.
REALLY NICE WORK.
AGAIN REMIND EVERYONE THAT IF
YOU HAVE QUESTIONS FOR THE
SPEAKERS, TO TYPE THEM ON THE
QUESTION AND ANSWER BOX AND
WE'LL ADDRESS THEM, THE SPEAKERS
CAN ADDRESS THEM DURING -- LIKE
BY TYPING, OR WE CAN ADDRESS
THEM AT THE END.
SO NOW WE'RE GOING TO MOVE ON TO
THE NEXT SPEAKER, AND IT'S A
PLEASURE FOR ME TO INTRODUCE DRI
BELIEVE RECENTLY MOVED TO ROACH
AND IS ROCHE
IN SWITZERLAND, BEFORE HE WAS
DIRECTOR AND PRINCIPAL
INVESTIGATOR IN THE NETHERLANDS,
ALSO INVESTIGATING DIRECTOR OF
RESEARCH AT THE CENTER FOR
PROCEED TRICK ONCOLOGY.
HE'SAND BEEN DOING PIONEERING
WORK IN ORGANOIDS, PARTICULARLY
INTESTINAL TRACT ORGANOIDS.
TODAY HE'S GOING TO BE TALKING,
THE TITLE OF THE PRESENTATION IS
HUMAN TISSUE STEM CELL DERIVED
ORGANOIDS MODEL VIRAL INFECTION.
HANS, THE FLOOR IS YOURS.
>> THANKS VERY MUCH.
, MARC, FOR THE INTRODUCTION AND
INVITATION TO SPEAK.
I'LL GIVE YOU A BRIEF
INTRODUCTION TO THE TYPES OF
ORGANOIDS THAT MY LAB HAS
DEVELOPED ABOUT 12 YEARS AGO AND
THEN I'LL GIVE YOU SOME EXAMPLES
OF HOW WE HAVE BEEN USING THIS
TECHNOLOGY TO MODEL VARIOUS
VIRAL INFECTIONS.
WHAT YOU SEE HERE IS THE INSIDES
OF THE HUMAN SMALL INTESTINE.
FOOD PASSES BY FROM LEFT TO
RIGHT.
THIS STRUCTURE HERE IS CALLED
THE VILLUS.
THERE'S ABOUT MAYBE 8 TO 10
CRYPTS THAT SURROUND THE BASE OF
THE VILLUS AND THESE
STRUCTURES -- WHEN WE WERE
STUDYING THE PATHWAY A LONG TIME
AGO BECAUSE THE STEM CELL
PROCESS THAT YOU SEE HERE, THE
CELERY NEWELL PROCESS
CELL RENEWAL PROCESS IS PROBABLY
THE FASTEST IN THE HUMAN BODY.
THEY PRODUCE DAUGHTER CELLS
EVERY DAY.
THOSE DAUGHTER CELLS TAKE ABOUT
TWO DAYS WHILE THEY DIVIDE THREE
OR FOUR TIMES TO EXIT THE
CRYPTS, THEY THEN MATURE INTO
SOMETHING LIKE 10 DIFFERENT CELL
TYPES THAT YOU NEED FOR
DIGESTING FOOD AND TAKING OF
NUTRIENTS.
THEY WILL THEN CONTINUE TO MOVE
UP AS FULLY DIFFERENTIATED CELLS
FIVE DAYS IN THE MOWRT, THEY
REACH THE TIPS OF THE VILLI AND
GO UP.
NOW THIS IS PROBABLY THE FASTEST
CELERY NEWELL
CELL SL RENEWAL PROCESS IN THE
MAMMALIAN BODY.
WE KNEW -- WAS A KEY TO DRIVE
THIS, AND THIS ALLOWED US TO
FINALLY IDENTIFY A MARKER FOR
THE STEM CELLS LGR5 IN MICE
WHERE WE COULD INSERT -- IN THE
LGR5 LOCUS, WE COULD EASILY SORT
OUT OF STEM CELLS.
WE NOTED LOTS OF UNEXPECTED
THINGS, THERE WERE MULTIPLE DOG
MASKS AROUND THE CELLS, THAT
THEY RARELY DIVIDE.
THESE STEM CELLS LIKE STEM CELLS
OF THE STOMACH OR THE HAIR
FOLLICLE, FOR INSTANCE,
CONSTANTLY DIVIDE, AND BASED ON
THIS OBSERVATION, TOSHIRO SATO
AND -- WE CAME UP WITH -- ONE WE
LATER TO FIND LIGAND OF LGR5 OF
OUR STEM CELL FACTOR, WE DO
THESE STEM CELLS IN MAT RIGEL IN
3D.
MUCH TO OUR SURPRISE, WE SAW
FORMATION OF THESE VERY
IRREGULAR STRUCTURES THAT UPON
CLOSE EXAMINATION TURN OUT TO BE
SMALL COPIES OF THE GUT
EPITHELIUM.
SO A SINGLE STEM CELL KNOWS HOW
TO BUILD THESE 3D STRUCTURES
WITH ALL OF THE DIFFERENTIATED
CELL TYPES, LINING THE CENTRAL
LUMEN WITH CRYPTS, AT THE BASE
OF THE CRYPTS ARE MULTIPLE STEM
CELLS AND THE OTHER CELLS THAT
RESIDE THERE.
SO REALLY REMARKABLE THAT THEY
CAN BUILD THIS, EVEN MORE
REMARKABLE THAT THEY CAN KEEP
GROWING AS THESE INTESTINAL
ORGANOIDS FOR MANY, MANY YEARS
AND WE'VE NEVER SEEN MUTATIONS
WHEN YOU TRANSPLANT THEM, THEY
ARE REALLY NORMAL TISSUE.
WE THEN REALIZED THIS PROTOCOL
WAS APPLICABLE TO PROBABLY ALL
EPITHELIA OF THE MAMMALIAN BODY.
YOU SEE A WHOLE LIST HERE.
WE HAVE PUBLISHED MULTIPLE
PROTOCOLS BUT I'M SURE MANY OF
THEM CAN STILL BE IMPROVED AND
THERE'S MULTIPLE OTHER PAPERS
THAT HAVE COME UP WITH
ADDITIONAL PROTOCOLS.
VERY BRIEFLY, YOU START YOUR
CULTURE NOT FROM A STEM CELL,
YOU JUST GET A SMALL PIECE OF
TISSUE.
THERE'S ALWAYS STEM CELL
ACTIVITY, BIOPSY OR SURGICAL
RESECTION OR FOR KIDNEY, FOR
INSTANCE, WE CAN JUST SPIN SOME
CELLS OUT OF URINE AND GROW THEM
HAD, YOU PUT THEM IN MAT RIGEL,
YOU ADD GROWTH FACTORS, KINASE
RECEPTORS, AND OFTEN YOU NEED
TO -- AND IF YOU FIND THE RIGHT
OPTIMIZED CULTURE CONDITION,
TYPICALLY YOU CAN GROW THIS
PIECE OF TISSUE PROBABLY
FOREVER.
VERY DIFFERENT FROM THE IPS
ORGANOIDS.
FROM THE MOMENT YOU PUT THEM IN
CULTURE, THEY'RE FULLY
SPECIFIED, THEY ACTUALLY PRODUCE
FULLY MATURE CELL TYPES BUT
THEY'RE VERY DONE, SO YOU CAN'T
TURN THEM INTO ANYTHING ELSE.
THEY'RE JUST GROW THE AREA OF
THE ORGAN OF INTEREST, THEY JUST
GROW THERE FOREVER IN THE FORM
OF ORGANOIDS.
ALSO WE CAN ONLY GROW EPITHELIAL
ELEMENTS FROM THESE TYPES OF
ORGANOIDS SO WE DON'T GET BONE,
WE DON'T GET MUSCLE, WE DON'T
GET BRAIN.
BUT RESPIRATORY AND INTESTINAL
VIRUSES TYPICALLY GO FOR
EPITHELIAL CELLS, ALL THESE
OTHER ORGANS ESSENTIALLY IN THE
EPITHELIAL CELL LAYERS OF THESE
ORGANS.
WHAT YOU CAN SEE HERE, STRAIGHT
FROM -- YOU CAN GROW CANCER
CELLS FROM A PATIENT, YOU CAN
GROW CELL FROM CYSTIC FIBROSIS
PATIENTS, LUNG, KIDNEY, STOMACH
AND RECAPITULATE PHENOTYPE AND
IT WILL REMAIN YOU THE EXAMPLES
OF INFECTIOUS DISEASE ORGANOIDS.
THIS IS AN ORGANOID THAT WE HAVE
WORKED VERY HARD AND VERY LONG
TO GET GROWING, HUILI HU IN
CHINA MANAGED TO COME UP WITH A
COMPLICATED COCKTAIL, EIGHT
GROWTH FACTORS.
IF WE HIT THE PROTOCOL RIGHT,PI
SME OF LIVER TISSUE WILL
GROW FOR LONG PERIODS OF TIME,
AND IT CONSISTS ENTIRELY OF
FULLY MATURE HEPATOCYTES THAT
OCCASIONALLY DIVIDE.
THEY'RE NOT THE FASTEST GROWER,
THEY PROBABLY GROW ABOUT THREE
FOLD HER WEEK, THESE CELLS ARE
FULLY MATURE HEPATOCYTES
DIRECTLY COMPARABLE TO PRIMARY
HE
HEPATOCYTES, SO CITE DIFFERENT
FROM -- ONE THING HERE, ANOTHER
CHINESE POSTDOC IN THE LAB,
BECAUSE WE CAN GROW THESE
ORGANOIDS FOR SUCH A LONG TIME,
WE CAN NOW STUDY CHRONIC
INFECTION BY VIRUSES AND THIS IS
HBV, THE CORE ANTIGEN OF HBV IS
MODELED HERE, AND THIS IS AFTER
32 DAYS IN CULTURE, AFTER
INFECTION, THEY CAN KEEP IT FOR
MONTHS.
YOU CAN SEE HIGH LEVELS OF THE
CORE ANTIGEN HERE IN GREEN, SO
THE VIRUS IS THERE, WE CAN SEE
FOR THE CC DNA -- AND THEY
SECRETE LIVE VIRAL PARTICLES AS
YOU CAN SEE IN THESE CURVES.
SO WE'RE WORKING TO MAKE
DISCOVERIES IN THIS MODEL SYSTEM
BUT IT SHOWS SOMETHING VERY HARD
TO STUDY PRIOR TO ORGANOID
TECHNOLOGY IF ONE GROWS IT AND
INFECTS IT WITH A VIRUS, ONE CAN
BUILD A CHRONIC MODEL FOR
INFECTION.
SO THIS IS CERVICAL ORGANOIDS,
SO LIVE CELLS TO GROW THESE
ORGANOIDS, AND SHE INFECTS WITH
HSV VIRUS, YOU CAN SEE HOW
RAPIDLY -- I THINK THIS MOVIE
RUNS OVER TWO DAYS OR SO -- HOW
RAPIDLY FIRST OF ALL THE VIRUS
PROPAGATES AND ALSO HOW IT KILLS
THE EPITHELIAL ORGANOIDS.
THESE ORGANOIDS DON'T HAVE
LUMENS, THEY COME FROM A
STRATIFIED EPITHELIUM, SO
TYPICALLY YOU SEE A VERY SORT OF
LIKE A -- LAKE LUMEN SKIN, THE
MOST MATURE CELLS ARE IN THE
INSIDE, AND THE BASAL CELLS WITH
THE PROLIFERATIVE LAYER IS ON
THE OUTSIDE.
EVERY ORGANOID YOU SEE HERE
INDIVIDUAL CELLS, WE STAINED FOR
THESE CILIA, THE BEATING CILIA
OF THE CILIATED CELLS, IN
BETWEEN THE OTHER CELL TYPES OF
THE BRONCHUS, AND HERE YOU CAN
ACTUALLY SEE, IT WAS VERY
SURPRISING WITH NORMAN SACS GROW
THESE ORGANOIDS ORIGINALLY, THE
SLIME THAT'S PRODUCED BY THE
GLOBUS CELLS -- IT'S MOVING AND
IT'S BECAUSE OF THE CILIATED
CELLS THAT ARE ALSO PRESENCE,
YOU CAN SEE THE CILIA NICELY
BEAT AND YOU MIGHT ALSO OBSERVE
THAT THEY BEAT IN A CONCERTED
WAY, SO THEY BASICALLY BEAT THE
WAY THEY SHOULD BEAT IN THE
AIRWAY SO THEY BASICALLY
SYNCHRONIZE WHAT THEY DO, AND
THIS MAKES THAT THE MUCUS IN THE
CENTER OF THESE ORGANOIDS SPIN.
THESE, WE'VE USED FOR A NUMBER
OF VIRUSES.
THIS IS JUST MAKING A MOVIE OF
AN ORGANOID WITH NO INFECTION.
OUR COLLABORATORS IN HONG KONG
HAVE USED THESE ORGANOIDS TO
STUDY INFLUENZA VIRUSES A NUMBER
OF YEARS AGO.
THE PAPER IS VERY POPULAR NOW
WITH THE SARS AND SARS-COV-2
STUDIES.
AND IT TURNS OUT THAT ONE CAN
USE THESE ORGANOIDS TO DEFINE
PATHOGENIC EFFECTS OF NEW
INFLUENZA STRAINS, PARTICULARLY
THE ONES THAT JUMP FROM PIG OR
BIRDS FROM HUMANS CAN BE VERY
DANGEROUS.
IN THE PAST WHEN THEY USED THIS
PLATFORM, THEY COULD ACTUALLY
SEE THIS IN CULTURES.
SO NOW IF WE ADD THE RSV VIRUS
ON THE NEXT MOVIE, HERE YOU SEE
THE ORGANOID AND SOMEHOW THEY
BECOME EXTREMELY MOTILE AND THEY
ACTUALLY BECOME CANNIBAL, THEY
EAT HEALTHY -- EAT AND AFFECT
HEALTHY ORGANOIDS.
YOU CAN SEE HERE.
THERE'S ANOTHER MOVIE HERE.
THIS ONE HERE.
YOU SEE THE FIRST IN RED, THE
FIRST RSV INFECTED ORGANOIDS, WE
ALSO SEE HUMAN LUNGS, THE
AFFECTED EPITHELIAL CELLS,
ORGANOIDS CONSIST ENTIRELY OF
EPITHELIAL CELLS, THEY VERY
RAPIDLY UNDERGO VMT AND WE
OBSERVE NOW IN THE LUNGS OF
INFECTED PATIENTS THAT THESE
CELLS HELP SPREAD THE VIRUS.
THE VIRUS HIDES INSIDE THESE
CELLS, WE BELIEVE, AND THEY
TRAVEL THROUGH THE AIRWAYS.
ANOTHER THING YOU CAN DO IS NOW
COINCUBATE WITH NEW CELLS AND
THESE ARE LYMPHOCYTE, NOTHING
MUCH HAPPENS WHEN WE DON'T
INFECT AND ON THE RIGHT, WE
TRACE INDIVIDUAL CELLS AND THEY
DON'T DO MUCH AS YOU CAN SEE.
BUT IF YOU INFECT THEM AGAIN BUT
NOW WITH A GREEN RSV VIRUS, YOU
CAN SEE THE MOMENT THEY GET
INFECTED, THE ORGANOIDS BECOME
EXTREMELY INTERESTING FOR THE
IMMUNE CELLS, THEY SE SECRETE
CERTAIN CHEMOKINES, THAT THEY
ATTRACT THE LYMPHOCYTES.
SO ONE CAN BUILD VIRAL -- IN
THESE ORGANOIDS BUT ON TOP OF
THAT, ONE CAN -- IMMUNE CELLS.
FINALLY, A SLIGHTLY LONGER STORY
THAT WILL BE PUBLISHED AT THE
BEGINNING OF THE PANDEMIC, WHEN
OUR INSTITUTE CLOSED DOWN, I
SHOULD SAY THAT ALL OF THIS
ACTUALLY COMES FROM THE
INSTITUTE WHERE I STILL ADVISE
MY ACADEMIC LAB.
NOT FROM -- SO HERE YOU SEE ACE
2 EXPRESSION IN THE GUT.
WE REALIZE THAT ACE 2, WHEN IT
WAS PUBLISHED TO BE THE RECEPTOR
FOR SARS-COV-2 2 IS EXTREMELY
HIGHLY EXPRESSED IN THE SMALL
INTESTINES.
THESE ARE VILLI OF THE SMALL
INTESTINE THAT YOU SAW EARLIER
IN THE ANIMATION.
MUCH HIGHER ACTUALLY THAN IN THE
RESPIRATORY TRACT, AND THERE'S
ALSO SOME INDICATIONS THAT THE
VIRUS NOT ONLY HIT LUNGS, THERE
ARE GI ESTIMATES, QUITE COMMON.
NAUSEA, STOMACH ACHES, DIARRHEA,
AND ALSO WHEN THE FIRST PCR
ASSAYS BECAME AVAILABLE FOR SARS
COV-2, PEOPLE DETECT VIRAL RNA
IN STOOL.
CAN BE PRODUCED IN THE LUNG BUT
ALSO IN THE GUT.
SO WE GOT -- INFECTION OF EVERY
ORGANOID, CROW INTO EXPERTS,
MART LAMERS, BART HAAGMANS.
WE GET A -- INCREASE OF LIVE
VIRUS PARTICLES, WE ALSO GET A
FOUR LOG INCREASE IN VIRAL RNA.
MUCH HIGHER THAN IN ANY OTHER
TISSUE THAT WE HAVE TESTED, ALSO
MUCH HIGHER, MUCH MORE
REPLICATION THAN IN THE AIRWAYS.
AND IF YOU FOLLOW THIS -- WE
STAIN IN WHITE FOR THE NUCLEAR
PROTEIN OF SARS-COV-2 2, SO SARS
ONE INFECTED ORGANOID AFTER 24
HOURS.
WE KNOW THE VIRUS HAS TO ENTER
FROM THE LUMEN, ACE 2 IS A
LUMINAL PROTEIN, I'LL GET INTO
THAT A LITTLE BIT LATER, AND
ALSO ARE AS EXPECTED THE VIRUS
ENTERS THE CELL FROM THE LUMEN,
ONCE IT'S REPLICATED, IT'S
RELEASED AGAIN INTO THE LUMEN OF
THE GUT, AND THE SAME THING WILL
HAPPEN IN THE RESPIRATORY TRACT.
IF WE WAIT THREE DAYS OR SO, WE
NOW SEE THAT IN AN AFFECTED
ORGANOID, ALMOST EVERY CELL IS
EFFECTED BUT THE NEARBY
ORGANOIDS DO NOT GET AIIVETTED ,
IT GETS EXTRUDED INTO THE LUMENS
AND CANNOT ESCAPE TO AFFECT
ANOTHER ORGANOID.
SO THIS WORKS.
WE DID A LOT OF SINGLE CELL
SEQUENCING AND THINGS WITH
INTERFERON, THE VIRUS BLOCKS
INTERFERON RESPONSES.
THAT WAS NOT TOO INTERESTING.
WE THEN WENT ON TO STUDY HOSTS
THAT THE VIRUS IS USING.
OBVIOUSLY IT HAS SO FEW GENES
ITSELF THAT IT MUST RELY HEAVILY
ON HOST GENES.
THE FIRST THING IS KNOCK OUT --
THESE ARE CLONAL ORGANOIDS SO WE
CONFIRM BY SEQUENCING THESE TWO
INDEPENDENT CLONES, WE HAVE A
NUMBER OF THESE WILD TYPE THAT
GETS HARD LIE AFFECTED WITH
SARS-COV-2, THE ACE 2 MUTANTS AS
EXPECTED DO NOT GET AFFECTED.
SO IT'S A GOOD CONTROL FOR WHAT
WE'RE TRYING TO DO HERE.
SARS, THE ORIGINAL VIRUS, IS
ALSO KNOWN TO USE ACE 2 AND
INDEED IT DOES NOT AFFECT WHEN
ACE 2 IS KNOCKED OUT.
MERS, A THIRD MEMBER OF THIS
CORONA DOESN'T CARE ABOUT THE
ABSENCE OF ACE 2 IN BLUE HERE
AND IT'S KNOWN TO USE ANOTHER
RECEPTOR, DPP4, AND INDEED WHEN
WE KNOCK OUT D PSM PP46789 NOWS
CANNOT INFECT BUT SARS-COV-2
2 CAN.
DPP4 -- WE CONFIRMED THAT FOR
THE GUT.
WHEN WE LOOK AT PUBLISHED
LITERATURE THERE ARE A NUMBER OF
GENOME-WIDE CRISPR SCREENS
REPORTED THAT HAVE COME UP WITH
DOZENS OF HUMAN GENES THAT WOULD
BE ESSENTIAL FOR VIRAL
REPLICATION.
WE KNOCKED OUT MOST OF THESE.
AGAIN, CLONAL, WE CONFIRM THAT
THEY CREATE NO ALLELES OF THESE
OR HOMOZYGOUS OF THESE GENES,
AND MUCH TO OUR SURPRISE, THE
ONLY GENE THAT WE CAN CONFIRM ON
TOP OF -- IN ADDITION TO ACE
2 IS TMPRSS2, SO THAT'S THE
SURFACE PROTEASE THAT CLEAVES
THE SPIKE PROTEIN AND ALLOWS THE
VIRUS THEN TO FUSE ITS ENVELOPE
WITH THE HOST MEMBRANE, BUT NONE
OF THESE OTHER GENES ACTUALLY
GAVE US ANY INHI INHIBITION Y
ANY INHIBITION.
ANOTHER OBSERVATION IS THAT
CHLOROQUINE, PROBABLY ALL OF
KNOW CHLOROQUINE,
HYDROXYCHLOROQUINE WERE VERY
POPULAR IN THE FIRST YEAR OR SO
OF THE PANDEMIC FOR SARS-COV-2
PATIENTS.
INDEED WE CAN REPRODUCE THAT IN
VERO E6 CELLS, AFRICAN MONKEY
CELLS FROM THE KIDNEY, THAT WHEN
YOU LOOK AT THEM LOOK MORE LIKE
FIBROBLASTS.
YOU TYPICALLY HAVE TO TRANSFECT
IN ACE 2 TO MAKE THEM RECEPTIVE
TO THE VIRUS.
BUT WHEN YOU DO THAT, YOU THEN
EFFECT WITH THE VIRUS AND ADD
CHLOROQUINE, CHLOROQUINE NICELY
BLOCKS INFECTION WITH AN IC50 OF
ONE MICROMOLAR AND 10
MICROMOLAR, VIRAL REPLICATION IS
ENTIRELY DEAD.
IF YOU NOTE THE SAME AMOUNT OF
CHLOROQUINE OR
HYDROXYCHLOROQUINE AND ADD TO AN
ORGANOID CULTURE, WE SEE NO
EFFECT WHATSOEVER, SO GREEN IS
THE WILD TYPE, BLUE IS THE
CHLOROQUINE-TREATED ORGANOID, SO
10 MICROMOLAR, NO EFFECT, IN
VERY SICK CELLS YOU HAVE FULL
BLOCKADE.
HOW TO EXPLAIN THIS, THERE ARE
OTHER PAPERS THAT SHOW THE SAME
PHENOMENA.
WHAT WE THINK IS HAPPENING IN
THESE NON-PRIMARY EPITHELIAL
CELLS THAT ARE OFTEN USED IN
VIROLOGY LABS IS THAT THE VIRUS
WILL BIND TO ACE 2 IF IT'S
PRESENT.
IT THEN GETS ENDO CYTOSED INTO
THE CELL AND FINALLY WHEN THE
RNA GENOME NEEDS TO BE RELEASED
FROM THIS PARTICLE HERE, THIS
INVOLVES THE CATH EP SINS THAT I
JUST SHOWED.
OF COURSE ENDO CYTOSIS IS
HIGHLY -- TO CHLOROQUINE AND
WOULD ALSO BE HIGHLY SENSITIVE
TO -- YOU HAVE AIRWAYS OR
INTESTINAL TRACT BUT ALSO
ORGANOIDS AND INTESTINAL TRACT
ORGANOIDS IS THERE'S A REAL
APICAL DOMAIN WHERE NATURALLY
ACE 2 IS EXPRESSED SO THE VIRUS
BINDS.
NATURALLY ALSO MOST -- CAN BE
KNOCKED OUT ALL -- THEY DO
NOTHING, IT'S ONLY TMPRSS2 THAT
HAS THAT EFFECT, THAT ACTUALLY
CLEAVE AND -- THE VIRAL ENVELOPE
DIRECTLY FUSES WITH THE HOST
MEMBRANE, THE VIRUS INFECTS ITS
RNA GENOME INTO THE HOST CELL
AND INFECTION IS STARTED.
THERE'S NO ENDO CYTOSIS UPON
INFECTION OF THE PRIMARY
EPITHELIAL CELL AND NO
SENSITIVITY TO HYDROXYCOLOR KIN.
SO LESSON TO BE LEARNED HERE IS
IT MAY BE GOOD TO INVOLVE
ORGANOIDS IN ADDITION TO THE
MORE EASY TO WORK WITH CELL
LINES IF I WANT TO STEP FROM
OBSERVATION IN THE LAB INTO THE
CLINIC.
AND WITH THAT, I THINK I NAMED
THE IMPORTANT INVESTIGATORS
ALREADY, COLLABORATORS ALREADY,
AND OF COURSE
THAT ENDS MY TALK, I THANK YOU
VERY MUCH FOR YOUR ATTENTION AND
I'LL STOP HERE AND GIVE THE
FLOOR BACK TO MARC.
>> THANKS, HANS.
BEAUTIFUL PRESENTATION.
I THINK WE ACTUALLY HAVE TIME --
WE'RE AHEAD OF SCHEDULE.
THERE'S A QUESTION ON THE CHAT
FOR YOU THAT SAYS THE VIDEO OF
THE RSV TOMATO-INFECTED 3D
ORGANOIDS LOOKS REALLY NICE.
WHICH MODEL OF MICROSCOPE DO YOU
USE FOR LIGHT IMAGING?
I GUESS WE KNOW SOME OF THESE
ORGANOIDS ARE HARD TO IMAGE.
RAPID LOW ENERGY CON FOCALS BUT
I DON'T KNOW HOW THIS MOVIE WAS
MADE.
>> GREAT.
THANKS SO MUCH, HANS.
APPRECIATE IT.
HANS, I BELIEVE YOU WON'T BE
ABLE TO BE IN THE QUESTION AND
ANSWERS, I DON'T KNOW HOW LONG
YOU CAN STAY.
>> I HAVE TO APOLOGIZE.
I HAVE TO STEP OUT IN A FEW
MINUTES.
>> YEAH.
SO THERE'S A FEW QUESTIONS YOU
CAN TAKE IF YOU HAVE TIME
OFFLINE.
BUT WE'LL CONTINUE NOW, AND THE
NEXT SPEAKER IS PROFESSOR LINDA
GRIFFITH.
SHE IS MIT SCHOOL OF ENGINEERING
TEACHING INNOVATION PROFESSOR OF
BIOLOGICAL AND MECHANICAL
ENGINEERING, AND DIRECTOR OF THE
CENTER FOR GYNEPATHOLOGY
RESEARCH AT MIT, WHICH IS AN
INSTITUTE THAT FOSTERS BOTH
BASIC AND CLINICAL RESEARCH IN
PATHOLOGIES OF THE FEMALE
REPRODUCTIVE TRACT.
AND SHE'S A MEMBER OF THE
NATIONAL ACADEMY OF ENGINEERS,
NATIONAL ACADEMY OF MEDICINE,
NATIONAL ACADEMY OF INVENTORS, A
IF LOW OF THE AMERICAN ACADEMY
OF ARTS AND SCIENCE.
SHE WAS A RECIPIENT OF THE
MACARTHUR FOUNDATION FELLOWSHIP.
AND THE GRIFFITH LAB HAS BEEN A
PIONEER IN USING TISSUE
ENGINEERING AND BIOMATERIALS
TECHNOLOGIES FOR REGENERATIVE
MEDICINE, DRUG DEVELOPMENT AND
UNDERSTANDING DISEASE
PATHOPHYSIOLOGY.
LINDA, THE FLOOR IS YOURS.
>> THANK YOU.
LET ME SHARE MY SCREEN.
SO I'M GOING TO SHARE WITH YOU A
VERY USEFUL TOOL, SO THERE'S ONE
MAIN THEME OF MY WORK, BUT I'M
GOING TO PUT AROUND THE
PRESENTATION THAT THE CONTEXT
FOR WHICH WE DEVELOPED THIS
TOOL.
SO BY WAY OF INTRODUCTION, JUST
BIG CONTEXT FOR MY LAB, WE'VE
BEEN INVOLVES IN ORGANS ON A
CHIP WORK SINCE THE BEGINNING OF
THE 1990s DEVELOPING VARIOUS
KINDS OF MODELS, AND AN
OVERARCHING CHALLENGE THAT WE
ADDRESS IN OUR WORK IS THE NEED
TO USE DEVICES THAT DO NOT
ABSORB LIPOPHILIC COMPOUNDS.
SO I'M SHOWING YOU A PROJECT WE
DID WITH DARPA SPONSORSHIP TO
PUT 10 ORGANS TOGETHER ON A CHIP
FOR A MONTH, AND THE REASON TO
SHOW THIS CHIP IS NOT THAT CUTE
AND ATTRACTIVE, BUT IT IS VERY,
VERY GOOD FOR PHARMACOKINETICS.
AND MY MAIN RESEARCH INTERESTS
ARE IN INFLAMMATION, SO RELEVANT
TO VIRUSES.
SO THE TOOLS I'M GOING TO TALK
ABOUT TODAY ARE MAINLY VALUABLE,
I THINK IN THIS COMMUNITY,
BECAUSE WE CAN CAPTURE
INFLAMMATORY EVENTS THAT ARE
REGULATED BY A LOT OF THINGS,
INCLUDING STEROID HORMONES,
WHICH ABSORB INTO PDMS.
SO AFTER THE BIG DARPA PROJECT,
WE'VE GONE ON TO DO A NUMBER OF
PROJECTS WITH THAT PLATFORM,
MAINLY MARTIN TRAPECAR, POSTDOC
LOOKING AT INFLAMMATION BETWEEN
THE GUT, LIVER AND BRAIN AND HOW
THAT'S MODULATED WITH SHORT
CHAIN FATTY ACIDS.
AGAIN, THAT PLATFORM IS NOT
HIGH-THROUGHPUT, IT'S NOT
INCREDIBLY PORTABLE TO OTHER
LABS, ALTHOUGH OTHER LABS HAVE
USED IT, BUT IT DOES HAVE THE
VALUE OF CAPTURING COMPLEX
ORGAN-ORGAN CROSSTALK AND
INFLAMMATION SO IT DOES HAVE
RELEVANCE FOR INFECTION.
NOW ALL OF THIS WORK SPARCED US
IN RECENT YEARS TO DO TWO BIG
THINGS, AND THAT IS TO DEVELOP
BETTER MODELS OF THE FISH TIS
THEMSELVES USING ORGANOIDS, SO
IT'S GREAT TO HAVE HANS AHEAD OF
ME, AND SECONDLY, TO PUSH REALLY
HARD TO MAKE BETTER DEVICES,
PARTICULARLY ONES THAT ARE
VASCULARIZED.
AND WE DO THIS IN THE CONTEXT OF
A DISEASE CALLED ENDOMETRIOSIS,
WHICH IS A COMPLEX INFLAMMATORY
DISEASE CHARACTERIZED BY ECTOPIC
GROWTH OF ENDOMETRIUM, WHICH IS
THE LINING OF THE UTERUS, SO YOU
CAN SEE HERE THESE LESIONS, THEY
CAN BECOME VERY INVASIVE, INVADE
THROUGH THE DIAPHRAGM AND GET TO
THE LUNGS.
THEY'RE BEAUTIFUL CYSTIC-LIKE
STRUCTURES, SINGLE EPITHELIUM,
JUST LIKE IN THE END ENDOMETR.
WHY IT CAN IB INVADE A MUSCLES
ANYBODY'S GUESS RIGHT NOW, BUT
IT DOES FORM THE CONTEXT FOR THE
ORGANOID WORK WE DO.
WHEN WE CAME IN THE FIELD, A
CONTRIBUTION WE MADE WAS TO
START EFFORTS TO DO MOLECULAR
STRATIFICATION OF THE DISEASE ON
THE BASIS OF INFLAMMATORY AND
INNOVATION SIGNATURES.
THERE'S NOT GREAT GENETICS FOR
THIS DISEASE, UNLIKE CANCER,
THERE AREN'T SOMATIC MUTATIONS,
SO WE HAVE BEEN INVESTIGATING
THE INFLAMMATION SIGNATURES IN
THE EU TO PICK TOPIC TISSUE.
ALL OF THEM HAVE FEATURES OF THE
EPITHELIAL, STROH MOLE AND
IMMUNE INTERACTIONS THAT ARE
ARRANGED.
I WANT TO FOCUS MORE ON THE
TOOLS BUT THIS IS REALLY
IMPORTANT CONTEXT.
A FIRST THING WE DID WAS A STUDY
IN ABOUT A HUNDRED WOMEN, WE
SHOWED THAT PERITONEAL FLUID HAD
SIGNATURES THAT ALLOWED A
MOLECULAR CLASSIFICATION OF
ABOUT A THIRD OF THE PATIENTS
WHO HAD ALL THE SURGICAL DISEASE
STAGES, AND WE USED THIS
MOLECULAR SIGNATURES WITH
BIOINFORMATICS TO PREDICT THAT A
CERTAIN KIND OF MACROPHAGE WAS
MAKING THESE CYTOKINES AND THAT
JUN KINASE WAS REGULATING IT.
FIRST TIME THERE WAS A REAL DRUG
TARGET THAT WASN'T A HORMONE.
JUN KINASE WAS ALSO IMPLICATED
IN AN INNOVATION STUDY WE DID,
COMBINING IN VITRO MODELS AND IN
VIVO SAMPLE, SO WE BECAME VERY
INTERESTED IN THIS AS A DRUG
TARGET AND WE LEARNED THAT THERE
HAD BEEN A CLINICAL TRIAL THAT
HAD FAILED BASED ON GREAT
PRE-CLINICAL DATA.
SO IN TALKING TO PHARMA, THEY
SAID WE NEED BETTER MODELS.
SO THIS IS REALLY WHERE I WANT
TO FOCUS.
SO TOGETHER WITH OUR WONDERFUL
CLINICAL COLLABORATORS, WE GET
SAMPLES FROM PATIENTS, AND BUILD
TISSUE BANKS, AND THESE ARE WELL
CLINICALLY PHENOTYPED PATIENTS.
AND OF COURSE THEN WE WANT TO
RECONSTRUCT AVATARS OF THE
PATIENTS THAT MAY LET US TEST
COMPLEX DISEASE MECHANISMS.
SO I'M GOING TO FOCUS HERE ON
BIDDING MODELS OF THE AIN'T
ACTIONS BETWEEN EPITHELIA,
STROMA AND IMMUNE CELLS AND
LATER VASCULAR TISSUE.
NOW EVERYTHING THAT HANS CLEVERS
JUST SPOKE ABOUT HAS BEEN
ENORMOUSLY ENABLING FOR LABS ALL
OVER THE WORLD WHO USE ANY KIND
OF MUCOSAL BARRIER TISSUES.
SHOWN BY TWO DIFFERENT GROUPS WE
WORKED WITH, THE TURCO, ET AL.
INVESTIGATORS, AND ALL THE
ENDOMETRIUM LABS AROUND THE
WORLD USE VERSIONS OF THESE.
NOW A HUGE PROBLEM AND SOMETHING
I WANT TO FOCUS ON, I THINK IS
USEFUL TO THE COMMUNITY, IS THAT
ALL OF THESE ORGANOIDS ARE
GENERALLY GROWN IN MAT RIGEL.
MOST GROW THEM IN -- CULTURE,
BUT ESPECIALLY FOR US BECAUSE
WE'RE TRYING TO BUILD COMPLEX
TISSUES, THE INTERACTIONS WITH
STROMAL AND IMMUNE CELLS ARE NOT
SO FAVORABLE.
IT ALSO HARD TO PUT IN DEVICES
AND DO ALL THE THINGS WE WANT TO
DO WITH TISSUE ENGINEERING.
SO WE UNDERTOOK A FAIRLY
COMPREHENSIVE EFFORT TO BUILD
SYNTHETIC HYDROGELS THAT COULD
BE USED FOR TISSUE ENGINEERING
OF THE MUCOSAL BARRIER
STRUCTURES AND THESE OF COURSE
ARE VERY IMPORTANT IN VIRUSES TO
INCLUDE IMMUNE CELLS PAUSE THE
INTERACTIONS WITH IMMUNE CELLS
OF COURSE ARE GOING TO DICTATE
WHAT HAPPENS.
WE INTRODUCED AN APPROACH
DECADES AGO USED ALL OVER THE
WORLD BUT WE CUSTOMIZED IT IN
VERY SPECIAL WAYS.
AT THE HEART OF THIS IS MULTIARM
PEGS MODIFIED WITH SMALL
PEPTIDES, WE TYPICALLY -- THESE
CAN BE BLADE INTO A GEL BY A.
PH CHANGE VERY SAFELY AROUND
CELLS.
WHAT WE DID THAT WAS SPECIAL AND
DRIVEN HUGELY BY OUR DESIRE TO
HAVE A MATRIX THAT THAT WOULT
NOT ONLY EPITHELIA BUT STROMA,
IMMUNE CELLS, VASCULAR CELLS, ET
CETERA, IS TO THINK ABOUT FIRST
THE INTERACTION CELLS WOULD HAVE
WHEN THEY'RE NAKED AND FIRST PUT
INTO THE GEL, THEY NEED TO
INTERACT WITH INTIGRINS TO
SURVIVE.
ONE OF THOSE THINGS THE DO
OBVIOUSLY IS TO DEPOSIT THEIR
OWN MATRIX AND REMODEL THEIR
MICROENVIRONMENT.
SO WE INCLUDE IN OUR GEL TWO
SPECIAL PEPTIDES THAT INTERACT.
I NOTE WE HAVE A DIFFERENT FORM
OF AN RGD INTERGREN.
A LOT OF FOLKS USE -- MOST CELLS
DOCTOR NORMAL CELLS DON'T HAVE
ALPHA B BETA 3 SO WE USE A
LIGAND THAT INTERACTS WITH OTHER
RGD INTERGRENS.
THEN WE ALSO HAVE PEPTIDES THAT
BIND TO EXTRACELLULAR MATRIX.
THAT CELLS THEMSELVES DEPOSIT.
SO WHERE THERE ARE FIBROBLASTS,
THE PROVISIONAL MATRIX OF
IVERMECTIN AS SHOWN HERE IS
STABILIZED SO CELLS CAN MAKE
MORE INTERACTIONS THROUGH THEIR
OWN DPOSTIVE MATRIX AND WHERE
THERE ARE EPITHELIAL CELLS THAT
DEPOSIT BASEMENT MEMBRANE, THAT
IS ALSO SEQUESTERED BY BINDING
PEPTIDES IN THE MATRIX.
AND WE FOUND THAT THOSE
FACILITATE THE ABILITY OF CELLS
TO BUILD THEIR OWN MICRO
ENVIRONMENTS AND FUNCTION AS
THEY WOULD IN THE TISSUE.
NOW IN THE LAB, EVEN THOUGH I'M
INCREDIBLY INTERESTED IN THE
ENDOMETRIUM, WE ALSO STUDY THE
GUT, AND DEVELOP TISSUE
ENGINEERING APPROACHES FOR THAT
AND A THERE'S SOME GOOD REASONS
FOR THAT.
MANY SCIENTIFIC REASONS BUT ALSO
SOME VERY PRACTICAL ONES.
THE PERSON WHO REALLY PUSHED THE
ORGANOID SIDE OF THIS INTO
EXISTENCE IS VICTOR
HERNANDEZ-GORDILLA WHO DID A
SCREAN GUIDED BY WHAT WAS KNOWN
ABOUT SOME OF THE ADHESIVE
INTERACTIONS AND THINGS GOING ON
IN THE STEM CELL NICHE OBVIOUSLY
IN WORK THAT HANS HAD PUBLISHED.
IN DOING THE SCREEN, HE LOOKED
AT A LOT OF DIFFERENT LIGANDS,
BIOPHYSICAL PROPERTIES,
BIOCHEMICAL PROPERTIES AND
IDENTIFIED A COMPLETELY TOTALLY
SYNTHETIC MATRIX SO THERE'S NO
LAMININ OR ANYTHING ADDED TO
THIS THAT ALLOWS CLONAL GROWTH
FROM EITHER THE UPPER OR LOWER
GI TRACT OVER TIME, AND IT'S NOT
AS EFFICIENT AS MAT RIGEL IN THE
ORIGINAL -- BUT IT DOES ALLOW
THE CLONAL GROWTH AND ALL THESE
OTHER FEATURES.
IT REQUIRES HAVING A PARTICULAR
LIGAND IN THE MATRIX AND SHOWN
HERE, IF YOU MAKE A
SUBSTITUTION, WE DON'T GET
ORGANOIDS.
STWA TO SHOW YOU THE IMPORTANCE
OF THIS FOR CELL-CELL
INTERACTION, I'M GOING TO COME
BACK TO THE ENT ME TREE UM.
IT FASCINATING BECAUSE IT GOES
THROUGH THESE ENORMOUS GROWTH
AND REGENERATION CYCLES EVERY 28
DAYS DRIVEN BY FIRST
PROLIFERATION UNDER THE PUSH OF
ESTROGEN, SO ESTROGEN DERIVES
ABOUT A COUNCILMEMBER GROWTH OF
THE TISSUE, THEN THERE'S
OVULATION AND PROGESTERONE COMES
IN AND CAUSES MATURATION OF THE
TISSUE SUCH THAT THE FIBROBLASTS
BECOME EPITHELIAL-LIKE OR AT
LEAST A SUBPOPULATION BECOMES
ACUTELY ?EA SENT.
SO WE WANT TO RECAPITULATE THIS.
WE KNOW MEN AND WOMEN DO NOT
HAVE EQUAL RESPONSES TO
INFECTION AND IMMUNITY, AND THIS
IS ACTUALLY A HUGE TOPIC WE
STARTED A DISCUSSION GROUP IN
THE BOSTON AREA AROUND THAT.
SO FOR PURPOSES OF THIS TALK,
PROGESTERONE INDUCES SOMETHING
CALLED A DECIDUALIZATION
RESPONSE.
YOU CAN SEE THEY BECOME PLUMP
LIKE EPITHELIA.
IT'S A VERY EASY ASSAY IN VITRO.
MANY UTERINE DISORDERS BEYOND
ENDOMETRIOSIS ARE CHARACTERIZED
BY PROGESTERONE RESISTANCE AND
CELLS THAT WILL NOT MAKE
PROLACTIN IN RESPONSE TO
PROGESTERONE.
SO IF WE CO-CULTURE NOW OUR
EPITHELIAL ORGANOIDS FROM
PATIENT END ME TREE A, THESE ARE
HEALTHY PATIENTS WITH STROMA, WE
CAN TAKE THEM THROUGH A
SIMULATED CYCLE AND THIS IS JUST
SHOWING MORPHOLOGY.
THEY WILL MAKE PROLACTIN.
IF WE ADD AN INFLAMMATION QUEUE
SUCH AS IL-1 BETA, THESE CELLS
BECOME INFLAMED-LOOKING.
THERE'S A FLATTENING OF THE
EPITHELIA, THE STROMA BECOME
ACTIVATED, AND OF COURSE WHAT WE
FIND IS IF WE GIVE PROGESTERONE
IN THE CULTURE, YOU GET
PROLACTIN BUT NOT WHEN YOU GIVE
IL-1 BETA.
IMPORTANTLY, THE MATRIX ITSELF
CAN BE USED TO MODULATE THE
BIOPHYSICAL PROPERTIES WITHOUT A
CHEMICAL QUEUE FOR INFLA NATION,
WE GET FEATURES OF LESION-LIKE
BEHAVIOR.
SO THIS IS AN ACTUAL PATIENT
LESION BY WHOLE SHEET
MICROSCOPY, YOU CAN SEE IT
INVADING IN THIS CASE INTO THE
MUSCLE, THE MYOMETRIUM, AND THIS
IS A SOFT MATRIX, IT HAS THE
PROPERTIES OF ENDOMETRIUM.
HERE'S THE MECHANICAL PROPERTIES
OF THE TISSUES HERE AND OF OUR
MATRICES.
SO IN THE SOFT PEG MATRIX, THE
EPITHELIAL STROMAL COCULTURES
LOOK BEAUTIFUL, BUT IN THE STIFF
MATRICES, THEY ACTUALLY TAKE ON
AN INFLAMMATORY PHENOTYPE.
NOW I WANT TO ILLUSTRATE WHY WE
ARE VERY EXCITED ABOUT THIS, AND
WE'VE SHARED THIS MATRIX WITH
MANY LABS NOW WHO ARE USING IT
FOR VARIOUS THINGS, BUT REALLY A
WONDERFUL COLLABORATION IS WITH
HILLARY CRITCHLEY, PROFESSOR OF
OGBYN IN EDINBURGH, SHE IS
PROBABLY THE WORLD EXPERT ON
HEAVY MENSTRUAL BLEEDING SO SHE
TREATS A LOT OF PATIENTS IN HER
CLINIC WITH SELECTED
PROGESTERONE RECEPTOR MOD LAY
FOR.
PROGESTERONE IS A PERFECT
AGONIST FOR IT.
AND MIFEPRISTONE, ABORTION
DWRUG, IS AN ANTAGONIST, AND IN
THE U.S., WE DO NOT HAVE
APPROVAL TO USE THIS DRUG FOR
HMB, IN PART BECAUSE THERE'S
THESE WEIRD FOR MOLGS THAT SHOWT
SHOW UP IN THE ENDOMETRIUM AND
THERE'S CONCERN ABOUT POTENTIAL
CONTRIBUTIONS TO ENDOMETRIAL CAN
SE WHICH IS ON
CANCER, WHICH IS ON THE RISE.
THERE'S INDICATIONS THAT IT
CAUSES DOWN REGULATION OF BOTH
PROLIFERATION AND DESYDIZATION,
GIVING RISE TO THESE WEIRD
PHENOTYPES.
SO THE CRITCHLEY LAB HAS ACCESS
TO PATIENTS AND THEY DID A
BEAUTIFUL STUDY IN PATIENTS WHO
WERE TAKING THE DRUG AND TOOK
ENDOMETRIAL BIOPSIES BEFORE,
DURING AND AFTER TREATMENT WITH
THE DRUG, FROM THE ENDOMETRIAL
BIOPSIES, DID A LOT OF MOLECULAR
OMICS AND THEN ALSO MAKE TISSUE
BANKS SO THAT YOU COULD GROW THE
ORGANOIDS AND STROMA TOGETHER.
SO ALEX TSOLOVA, THE GRADUATE
STUDENT SHOWN BEAMING BECAUSE
SHE WON AN AWARD IN MARCH THIS
YEAR AND SHE JUST DEFENDED HER
THESIS AND THIS IS A PAPER IN
PREPARATION, SHE ASKED THE
QUESTION ABOUT CELL-CELL MATRIX
AND THE MECHANISM OF ACTION OF
SPRM SINCE SHE FIRST SHOWED YOU
HAVE TO HAVE THE CELLS
CO-CULTURED TOGETHER TO GET
ANYTHING LIKE AN IN VIVO
RESPONSE.
SO SHE'S GOT THE IN VIVO OMICS
DATA AND NOW SHE CAN COMPARE HER
CULTURES.
BUT A CRUCIAL FEATURE, I'LL JUST
SHOW YOU THE DATA HERE, IS
COMPARING MATRIGEL AND PEG FOR
THESE CO-CULTURES.
SO I'M GOING TO WALK YOU THROUGH
THIS BECAUSE IT'S REALLY
COMPLICATED.
WHAT SHE DID IS REVERSE PHASE
PROTEIN ARRAY OF -- REVERSE
PHASE ANTIBODY ARRAY WITH 100
ANTIBODIES CHOSEN FROM THE IN
VIVO OMICS DATA.
THAT DATA SHOWED THAT THE
SPRMs DOWNREGULATED A HUGE SET
OF GENES ASSOCIATED WITH CELL
PROLIFERATION.
SO FIRST LET'S LOOK AT PEG-GROWN
CELLS HERE.
ESTROGEN, THE RED GENES ARE ALL
PROLIFERATION GENES AND I'LL
JUST FOCUS ON THOSE FOR NOW.
YOU CAN SEE THAT ESTROGEN
UPREGULATES THOSE GENES AS YOU
WOULD EXPECT IN VIVO, AND THE
SPRM IN THE PEG SYNTHETIC
CO-CULTURE COMPLETELY
DOWNREGULATES THEM.
SO THIS IS SIMILAR TO WHAT WE
SEE IN VIVO FROM THOSE PATIENTS.
MATRIGEL IS ALL OVER THE MAP.
IT'S REALLY A MESS.
IT DOES NOT RECAPITULATE THE IN
VIVO.
SHE'S FOLLOWED UP WITH IHC AND
VARIOUS OTHER THINGS TO VALIDATE
THESE FINDINGS.
SO THIS IS AN AMAZING OUTCOME.
THE TAKE-HOME MESSAGE FROM THIS,
THEN I JUST HAVE A COUPLE MORE
SLIDES HOW WE'RE TAKING THIS
INTO A BIOREACTOR, IS THAT THE
SYNTHETIC MATRIX IS A LOW NOISE
MICROENVIRONMENT IN WHICH HE CAN
LISTEN TO THE CROSSTALK BETWEEN
THE DIFFERENT CELL TYPES.
NOW I DIDN'T SHOW IMMUNE CELLS
BUT THERE'S A SEPARATE
PUBLICATION WE HAVE RECENTLY
APPLYING TO PANCREATIC ORGANOID
STROMA IMMUNE CROSSTALK WHERE
THERE'S A LOT OF DETAILS.
SO IT'S LIKE YOU'RE CAPTURING
THE CELLS TALKING TO EACH OTHER
WITH NO BACKGROUND NOISE, LIKE
YOU'RE IN A QUIET QA FAY.
CAFE.
BUT MATRIGEL, IF YOU'RE TRYING
TO CAPTURE WHAT THE CELLS ARE
SAYING, IT'S LIKE YOU'RE ON THE
RUNWAY AT LOGAN AIRPORT AND JETS
ARE TAKING OFF, THEY CAN'T TALK
TO EACH OTHER VERY WELL.
SO THAT'S THE TAKEHOME.
WE'RE TRYING TO COMMERCIALIZE IT
IN THE ERA OF COVID, THERE'S
BEEN A LOT OF LAWYER DRAMA, SO
HOPEFULLY WE'LL BE ABLE TO GET
THIS OUT TO MORE LABS THAN THE
LABS WE'RE CURRENTLY
COLLABORATING WITH.
I'M GOING TO END BY SAYING OF
COURSE WE'RE TAKING THIS BACK IN
VIVO TO LOOK AT INTERACTIONS
BETWEEN THESE LESIONS AND BLOOD
VESSELS, AND I WAS THRILLED TO
SEE CHRISTINE MIMMERY SHOW THE
IMMUNE CHIP, AN IMPORTANT
FEATURE OF THESE KINDS OF
VESSELS IS YOU CAN ADD I MEUP AE
CELLS AND LOOK AT TRAFFICKING.
THIS IS JUST A GRAPHIC, THE
FIRST EXPERIMENT, OUR STUDENT
ELLEN DID IN THE AIM CHIPS.
BUT WHAT WE'VE BEEN DOING, ALL
OF THESE RELY ON USING FIBRIN
FOR FORMING THE BLOOD VESSELS.
IT BEEN HARD TO GET A SYNTHETIC
MATRIX.
WE'VE BEEN WORKING WITH ROGER'S
LAB TO BUILD OUR LESION MODEL,
AND FOR NOW, THERE'S SOME PDMS
DEVICES HE USED FOR TUMORS, AND
WHAT WE'VE BEEN ABLE TO SHOW IS
GROWTH OF VESSELS FROM A
PREFORMED VESSEL EMBEDDED
FIBRIN, THIS IS A PROTO LESION
IN PEG BECAUSE OUR LESIONS HAVE
TO BE GROWN IN BEG.
PEG.
FIBRIN DOES NOT WORK WELL FOR
OUR EPITHELIAL STROMAL
CO-CULTURES.
HOWEVER, WE HAVE A MANUFACTURING
PROCESS NOW IN PLACE TO TAKE
THESE CHIPS AND PUT THEM INTO
THERMOPLASTICS, INCLUDING
EMBEDDED PUMPS.
I'M NOT SHOWING THAT BECAUSE OF
SOME I.P. ISSUES, BUT THIS IS
WHERE WE'RE GOING.
SO THESE KINDS OF SYSTEMS COULD
BE VALUABLE FOR LOOKING AT VIRUS
INFECTION BECAUSE THEY SHOULD BE
AMENABLE TO CONTROLLING BOTH THE
BIOCHEMICAL AND THE BIOPHYSICAL
MICROENVIRONMENT, AND THERE'S A
LOT OF PEOPLE WHO CONTRIBUTED TO
THIS, ESPECIALLY DAVE TREMPER ON
THE DEVICE SIDE, HEAVY LIFTING
ONCOORGANOID CULTURE, HILLARY'S
LAB, ALEKS AND OUR FABULOUS
CLINICAL COLLABORATORS FOR THE
SAMPLES.
>> THIS IS GREAT.
THANKS SO MUCH, LINDA.
IT IS INTERESTING, THE SAME --
CHRISTINE AND YOU STARTING WITH
ORGANOIDS AND MICRO TISSUES AND
THEN AT THE END MOVING INTO THIS
VASCULARIZED BED TISSUE CHIP
PLATFORMS TO SORT OF MOVE THEM
INTO A MORE LIKE PERFUSE SYSTEM.
IT SEEMS TO BE LIKE A TREND OF
BLENDING TECHNOLOGIES INSTEADING
STUDYING SEPARATE BUT THEN
PUTTING THEM TOGETHER WHICH I
THINK IS FASCINATING AND
HOPEFULLY SOMETHING WE CAN
DISCUSS MORE AT THE END.
THANK YOU SO MUCH, LINDA.
>> THANK YOU.
>> WE'RE GOING TO MOVE TO THE
NEXT SPEAKER, AND IT'S PROFESSOR
ANTHONY ATALA, WHO IS ES THE
LINK PROFESSOR AND DIRECTOR OF
THE WAKE FOREST INSTITUTE FOR
REGENERATIVE MEDICINE AND THE
BOYCE PROFESSOR AND CHAIR OF THE
DEPARTMENT OF UROLOGY AT WAKE
FOREST UNIVERSITY.
DR. ATALA IS A MEMBER OF THE
NATIONAL ACADEMY OF MEDICINE AND
THE NATIONAL ACADEMY OF
INVENTORS, AND HE'S RECIPIENT OF
THE U.S. CONGRESS FUNDED
COLUMBUS FOUNDATION AWARD AND
THE EDISON SCIENCE AND MEDICAL
AWARD, AND R & D INNOVATOR OF
THE YEAR AWARD AND SMITHSONIAN
INGENUITY AWARD AMONG SOME
OTHERS.
AND DR. ATALA HAS BEEN A PIONEER
IN THE FIELD OF TISSUE
ENGINEERING, BIOPRINTING AND
ESPECIALLY GROWING TISSUES AND
ORGANS FOR PATIENTS AND
PHYSIOLOGICAL MODELING.
SO TONY, THE FLOOR IS YOURS.
>> THANK YOU, MARK.
IT'S GREAT TO BE WITH YOU AND
GREAT TO BE WITH THIS PANEL.
THANK YOU FOR PUTTING THIS
TOGETHER ALONG WITH YOUR TEAM.
WHAT I THOUGHT I WOULD DO TODAY
IS REALLY GIVE YOU AN OVERVIEW
OF SOME OF THE WORK WE'RE DOING
IN THE AREA OF THE BODY ON A
CHIP.
OUR AREA OF EXPERTISE OVER THE
LAST SEVERAL DECADES HAS BEEN
REGENERATIVE MEDICINE, BASICALLY
MAKING TISSUES FOR PATIENTS, AND
OF COURSE AS YOU CAN IMAGINE,
WE'RE MAKING TISSUES FOR
PATIENTS, WE HAVE TO GO THROUGH
THIS WHOLE APPROACH OF CELL IS
HARVESTING, CELL PROCURING,
EXPANSION OF CELLS OUTSIDE THE
BODY, MIXING THAT WITH BIOFEERLS
SO
BIOMATERIALS SO WE CAN PUT THEM
INTO PATIENTS.
SO REALLY THE STRATEGY FOR US
REALLY EARLY ON WAS HOW DO WE
MINIATURIZE THE SYSTEMS THAT WE
ARE DOING IN REGEN MED.
WE PUT A WHITE PAPER OUT ON THIS
INITIALLY BACK IN THE EARLY
2000s.
THE STRATEGY HERE WAS TO GO
AFTER THE SYSTEM IN A COMBINED
WAY, AND THAT WAS MADE POSSIBLE
BY A GRANT FUNDED BY DTRA, THE
DEFENSE THREAT REDUCTION AGENCY
THAT THEY ANNOUNCED IN 2012 WITH
OUR PARTNERS IN SEVERAL CENTERS
IN THE U.S.
REALLY THE MAIN GOAL OF OUR
PROGRAM WAS TO CREATE THESE
MICRO CHIP TECHNOLOGIES BUT TO
DO SO WITH BIOPRINTED HUMAN
TISSUES AS MUCH AS POSSIBLE, SO
WE STARTED PRINTERS THAT WE
DESIGNED OVER A 20-YEAR PERIOD
TO CREATE TISSUES AND ORGANS.
THESE PRINTERS WERE ABLE TO
STANDARDIZE THE SYSTEM AND ALLOW
THESE STRUCTURES TO BE PRINTED
IN THE RELIABLE MANNER THAT CAN
BE EXPANDED OVER TIME.
BASICALLY AS WE ALL KNOW WE'RE
PREACHING TO THE CHOIR HERE BUT
JUST TO REMIND EVERYBODY WHY
THIS, WHY DTRA CAME OUT WITH
THIS PROGRAM, MAINLY BECAUSE AS
YOU KNOW, DRUG DEVELOPMENT IS
CHALLENGING, AND IT TAKES TENS
OF MILLIONS OF DOLLARS TO GET A
TARGET TO PHASE 1 HUMAN CLINICAL
TRIAL, BUT UNFORTUNATELY 90% OF
THE DRUGS THAT REACH PHASE
1 CLINICAL TRIALS END UP
FAILING.
AND THAT REALLY HAS TO DOITH
THE SYSTEMS THAT ARE BEING USED,
RIGHT?
BASICALLY TO 2D CULTURES ARE T
REALLY 3D TISSUES AND ANIMALS
ARE NOT ALWAYS PREDICTIVE OF
HUMAN OUTCOMES.
THEREFORE, WITH THIS PROGRAM
THAT WE FIRST DEVELOPED WITH
DTRA, THE EX VIVO CONSOLE OF
HUMAN ORGANOIDS AND WE ASSIGNEDD
ORGANOIDS THE SAME -- WE USE THE
NORMAL HUMAN CELL TYPES THAT ARE
PRESENT IN THE ORGAN IN THE SAME
PROPORTION, SO THE LIVER HAS
FIVE DIFFERENT CELL TYPES, EACH
CELL TYPE HAS A ROLE THAT PLAYS
INTO THE FUNCTIONALITY, SO THESE
ARE INCORPORATED.
WE ALSO INCORPORATE THE
EXTRACELLULAR MATRIX WHENEVER
POSSIBLE, WHERE WE TAKE AN OR
GAL THAT WE DECELLULARRIZE, WE
THEN CREATE A POWDER TO CREATE
THE ORGANOIDS, SO BASICALLY ALL
THE ELEMENTS PRESENT IN THE
ORGAN, ALL THE CELLS AND THE
EXTRACELLULAR MATRIX.
WE EVEN CONTROL FOR THE
STIFFNESS BECAUSE THAT ALSO
INFLUENCES THE LOCAL
MICROENVIRONMENT.
THE OUTCOME IS STRUCTURES THAT
REALLY HAVE A HIGH FIDELITY TO
THE HUMAN, FOR EXAMPLE, WITH THE
LIVER AS YOU CAN EVEN SEE
PRIMITIVE BILIARY SYSTEMS THAT
YOU SEE HERE.
WHEN YOU LOOK AT THESE, THEY CAN
PRESERVE THEIR FUNCTION LONG
TERM.
THIS IS LOOKING AT THE LIVER,
ORGANOIDS LOOKING AT -- ALBUMEN
SECRETION.
FOR EXAMPLE, WHEN WE PUT THESE
INTO LIVERS WITH DRUGS LIKE
VALIUM, WE SEE IT GETS BROKEN
DOWN INTO ITS NORMAL METABOLITES
WITH THE UPREGULATION OF THE
RIGHT -- AS YOU WOULD EXPECT IN
A HUMAN.
SAME MANNER, WE CREATE THE
MINIATURE HEART ORGANS IN THE
SAME MANNER USING ALL THE MAJOR
HEART CELL TYPES IN THE SAME
PROPORTION.
WE CAN SLOW THEM DOWN OR SPEED
THEM UP FROM THEIR 60 BEATS PER
MINUTE BASED ON DIFFERENT DRUGS
AND THE HUMAN DOSE-RELEVANT
RESPONSE.
WE CAN ALSO CREATE MINIATURE
HEART ATTACKS, SO CREATE MODEL
SYSTEMS THAT ARE ABLE TO HELP US
OUT WHEN WE'RE LOOKING AT DRUG
DEVELOPMENT OR MODEL SYSTEMS.
WE CAN CREATE THESE MINIATURE
BRAINS.
SAME THING WITH THESE MINIATURE
BRAINS, WE USE ALL THE SIX MAJOR
TYPES THAT ARE PRESENT IN THE
BRAIN IN THE SAME PROPORTION,
AND BY DOING SO, WE'RE ABLE TO
CREATE AN AN ENTIRELY NATIVE N
BLOOD BRAIN BARRIER, THAT IS, A
BARRIER CREATED BY THE CELLS
THEMSELVES.
WE'RE NOT USING TRANS MEMBRANES
OR SILICONE MEMBRANES FOR THIS.
BLOOD BRAIN BARRIER IS CREATED
ENTIRELY BY THE MINIATURE BRAIN
ORGANOIDS THAT YOU SEE HERE,
WHERE WE CAN CREATE ALSO MIMIC
STROKES, FOR EXAMPLE, AND THEN
GO AFTER TREATMENT MODALITIES.
SAME THING FOR THE LUNG, THIS IS
A NICE SLIDE BECAUSE IT SHOWS
WHAT A 2D CULTURE SYSTEM LOOKS
LIKE ON THE TOP RIGHT, WHAT A
NATIVE LUK LOOKS LIKE ON E
BOTTOM RIGHT.
WE CAN SEE THEN WE CAN START
DOING INFECTIVITY.
FOR EXAMPLE, WE TOOK STRAINS OF
PSEUDOMONAS THAT WERE SHOWN TO
BE DIFFERENTIAL IN TERMS OF
THEIR AGGRESSIVENESS IN HUMANS
AND WE WERE ABLE TO TEST THAT IN
THE ORGANOID SYSTEM REPLICATING
BASICALLY THE INFECTIVITY
AGGRESSIVENESS THAT WE WOULD SEE
IN A HUMAN MODEL, SHOWING -- TO
THE HUMANS.
THIS IS LOOKING AT RSV, WHERE WE
CAN ACTUALLY INCORPORATE
NEUTROPHILS.
ON THE LEFT YOU CAN SEE THE
MODEL -- BUT ON THE TOP RIGHT,
YOU SEE WHAT HAPPENS WHEN WE
INFECT THE LUNG ORGANOIDS WITH
RSV, SHOWING MIGRATION OF THE
NEUTROPHILS TO THE EPITHELIAL
MEMBRANE WITH DISRUPTION OF
CILIA, AND YOU CAN SEE THE
INCORPORATION OF THE NEUTROPHILS
WITH THE IMMUNOFLUORESCENCE THAT
YOU SEE HERE.
WE CAN ALSO LOOK AT ZIKA VIRUS,
FOR EXAMPLE.
THIS WAS DONE IN TESTICULAR
TISSUE, SHOWING HOW WE CAN
ACTUALLY INFECT THESE WITH THE
ZIKA VIRUS AND LOOK AT THE
NATURAL HISTORY OF THE VIRUS
OVER TIME.
WE ALSO LOOKED AT COVID.
AS YOU CAN SEE HERE WITH THE
COVID INFECTION, WE CAN SHOW
THAT THE LUNG IS POSITIVE FOR
ACE 2, AND THEN YOU CAN SEE THE
SARS SPIKE PROTEIN WITHIN THE
ORGANOIDS AND YOU CAN SEE THE
EXPANSION OF THE VIRUS AS YOU
WOULD SEE WITH HEIGHTENED
REPLICATION AT DAY # 3 TO 4-RBS
WHICH IS WHAT YOU H
YOU WOULD SEE IN HUMANS.
-- MAXIMIZED AT DAY 4 ALSO WHAT
YOU WOULD SEE IN A HUMAN MODEL.
SO THESE REALLY DO RESPOND TO
THESE MODELS VERY WELL.
WE'RE NOT LOOKING AT THE TRAN
SCRIP TOE MINK ANALYSIS
TRANSCRIPTOMIC ANALYSIS OF
DRUGGABLE TARGETS, LOOKING AT
POTENTIAL DRUG TARGETS FOR THE
FUTURE.
AND ALSO LOOKING AT, FOR
EXAMPLE, WE DID USE OUR SYSTEM
TO TEST ANTIVIRAL AGENTS DURING
THE COVID PANDEMIC, AND ONE OF
THESE AISHTS IS NOW IN ENTRY
PHASE 2 CLINICAL TRIALS SHOWING
THIS AGENT WAS EFFECTIVE AGAINST
THE SARS-COV-2 VIRUS.
NOW WHAT DO THEY ACTUALLY LOOK
LIKE?
THIS IS AN ACTUAL PRINTED CHIP.
SO WE PRINT BOTH THE CHIP AND
THE ORGANOIDS TOGETHER, AND THIS
PARTICULAR ONE, WE HAVE LIVER,
KIDNEY AND OVARY ALL PRINTED
TOGETHER.
SO THE WHOLE STRUCTURE WAS
PRINTED HERE, THE CHIP AND THE
CELLS.
THE ONLY THING THAT WAS NOT
PRINTED WAS THE TUBING AND THE
GLASS SLIDE.
BUT EVERYTHING ELSE WAS PRINTED.
AND YOU CAN SEE HERE THAT WE CAN
PUT SIX TOGETHER.
IN THIS PARTICULAR ONE, WE HAVE
LIVER, HEART, INTESTINE, LUNG,
BLOOD VESSELS AND TESTES, WE CAN
HOOK UP 12 OF THESE TOGETHER IN
THE SAME SYSTEM.
AND WE'RE STARTING TO SEE NOW
THINGS THAT WE COULDN'T SEE
BEFORE.
SO FOR EXAMPLE, BLEOMYCIN IS
KNOWN TO BE TOXIC TO THE LUNG.
AND THAT'S BEEN KNOWN FOR
DECADES.
AND SO TO OUR SURPRISE, WHEN WE
PUT IT IN OUR SYSTEM, THE HEART
STOPPED BEATING AND WE COULDN'T
FIGURE OUT WHY THE HEART STOPPED
BEATING BECAUSE THERE'S NOTHING
IN THE LITERATURE TO SUGGEST
THAT THESE ORGANOIDS WOULD STOP
THE HEART -- THESE HEART
ORGANOIDS WOULD BE AFFECTED.
SO WE PUT A NEW BATCH IN.
SAME CHALLENGE, THEY AGAIN
WERE -- THEY STOPPED BEATING, SO
WE THOUGHT MAYBE IT WAS A LIVER
METABOLITE.
WE PULLED THE LIVER OUT, THEY
STILL STOPPED BEATING.
TURNS OUT THAT WHEN THE LUNG
STARTS TO BE TOXIC FROM THE
BLEOMYCIN, THAT THERE'S
SECRETION OF INTERLEUKINS AND
THE INTERLEUKINS, IN FACT, CAN
BE CARDIOTOXIC, SO THE FACT IS
THAT WITH SUBCLINICALLY, THIS IS
PROBABLY OCCURRING, WE DON'T SEE
IT BECAUSE BY THE TIME WE PULL
THE DRUG, THIS CHEMOTHERAPY
AGENT THAT IS COMMONLY USED, BY
THE TIME WE PULL IT FROM THE
PATIENT, YOU PROBABLY DO HAVE
CARDIAC SUBCLINICAL TOXICITY
GOING ON.
WE'RE NOW STARTING TO SEE THINGS
THAT BEFORE WERE NOT POSSIBLE TO
SEE.
HOW DOES THE SYSTEM WORK
ACTUALLY IN TERMS OF DRUGS?
SO WE TOOK ABOUT 40 DRUGS THAT
WERE RECALLED BY THE FDA.
I'M GOING TO GIVE YOU A COUPLE
EXAMPLES HERE, WHERE THERE ARE
DRASTIC DIFFERENCES.
THIS IS HISMANAL, ANTIHISTAMINE
THAT HAD BEEN USED IN THE
MARKETPLACE FOR 11 YEARS.
THE DRUG HAD BEEN TESTED IN THEY
WILL LINES, NO TOXICITY, ANIMAL
MODELS, NO TOXICITY.
HUMAN CLINICAL TRIALS, NO
TOXICITY.
GETS RELEASED INTO THE
MARKETPLACE, 11 YEARS LATER,
THEY FINALLY CONNECT THE DOTS
FOR THIS OVER THE COUNTER DRUG
TO SHOW THIS WAS ACTUALLY
CAUSING HEART BLOCK.
THE DRUG WAS RECALLED BY THE
FDA, WE TRUSTED THE DRUG, IN TWO
WEEKS WE KNEW THE DRUG WAS TOXIC
TO THE HEART, SAME RESPONSE IN
HUMAN.
THIS IS MORE OF A TRAGIC STORY.
THIS IS REZULIN, AN ANTIDIABETIC
ON THE MARKETPLACE FOR THREE
YEARS.
SAME, HAD BEEN TESTED IN CELL
LINES, ANIMAL MODEL, PHASE 1,
2 AND 3 HUMAN CLINICAL TRIALS,
ON THE MARKET FOR THREE YEARS
AND BASICALLY WHAT WE SAW WAS A
MAJOR CHALLENGE WITH 63 DEATHS
DUE TO LIVER FAILURE WITHIN
THREE YEARS.
SO WE WERE ABLE TO GET THE DRUG
TESTED IN OUR SYSTEM, SAME HUMAN
DRUG DOSE-RESPONSE.
SO THE SYSTEM IS FAIRLY USEFUL
WHEN LOOKING AT THESE DRUGS.
WE'RE NOW USING THIS FOR CANCER.
WE HAVE ABOUT 12 DIFFERENT
CLINICAL TRIALS GOING ON WHERE
WE TAKE TISSUES FROM PATIENTS
HERE AT WAKE FOREST WITH TUMORS,
WE CREATE A TUMOR IN A CHIP, WE
CAN THEN USE CHEMOTHERAPY AGENTS
TO LOOK AT PREDICTIVE MODELING
OF WHAT WOULD BE THE BEST
TREATMENT FOR THE PATIENTS IN
THE FUTURE.
SO THIS IS ACTUALLY ACTIVELY
ONGOING.
WE HAVE SEVERAL PROGRAMS RIGHT
NOW, WE HAVE A PROGRAM FROM
BARDA LOOKING AT CHEMICAL
AGENTS, A PROGRAM FROM DTRA
LOOKING AT NERVE AGENTS AND A
RECENT PROGRAM, WE'RE GOING TO
BE LOOKING AT VIRUSES.
SO BASICALLY HERE WHAT WE DO IS
WE ARE LOOKING AT A SELECTION OF
RELEVANT ORGAN TISSUE
EQUIVALENTS.
WE CAN EXPOSE THEM TO THE
VIRUSES OF INTEREST.
WE CAN THEN ANALYZE FOR
BIOMARKERS AND THEN WE CAN LOOK
AT A WIDE RANGE OF DOSES AND
RESPONSES FROM MILD TO SEVERE,
AND THEN BASICALLY WE'RE LOOKING
AT ALL THE OMICS.
SO CURRENTLY FOR EXAMPLE FOR
CHEMICAL PROGRAM AND SAME THING
FOR VIRAL PROGRAM, WE'LL BE
LOOKING AT TRANSCRIPTOMICS,
PROTEOMICS, METABOLOMICS.
WE LIKE TO SAY THERE ARE NO
OMICS LEFT BEHIND.
BUT THIS IS BASICALLY BEING DONE
SO WE CAN APPLY THESE CAPABILITY
TO GENERATE HIGH QUALITY AND
REPRODUCIBLE DATASETS FOR
DOWNSTREAM INTEGRATED
BIOINFORMATIC AND STATISTICAL
ANALYSIS SO WE CAN THEN IN THE
FUTURE TAKE A KNOWN VIRUS, LOOK
AT HOW THE VIRUS REPLICATE, LOOK
INTO BIG DATA, LOOK AT THE
BIOMARKERS, USE ARTIFICIAL
INTELLIGENCE AND MACHINE
LEARNING, SO WE CAN LOOK AT
EXPOSURE AND PROGNOSTIC
BIOMARKERS, LOOK AT PROPHYLAX S
AND THERAPEUTICS.
WE'RE USING THIS SYSTEM FOR
IDENTIFYING AND CREATING NEW
COMPOUNDS.
WE ARE NOW APPROACHING THROUGH
THE PATHWAY OF CLINICAL
TRANSLATION.
AND OF COURSE WITH THE SYSTEMS,
THERE'S STILL WORK TO BE DONE.
WE HAVE TO LOOK AT RELIABLE CELL
SOURCING, WE HAVE TO LOOK AT
INFECTIVITY STANDARDS,
REPRODUCIBILITY, AND GENETIC
VARIABILITY SO THERE'S STILL A
LOT OF WORK THAT NEEDS TO BE
DONE, BUT CERTAINLY IT'S A
SYSTEM THAT CAN LEND ITSELF TO
BEING USEFUL.
WE DO HAVE THIS CONSORTIUM
THAT'S LOOKING AT DEVELOPING
THIS AI MACHINE LEARNING
CAPABILITIES TO ACCELERATE DRUG
SCREENING AND DISCOVERY, AND
LOOKING -- OUR FOUNDING MEMBER
WAS ORACLE, SO THEY CAN -- TO
HELP MANAGE ASPECTS RELATED TO
THE A.I. FOR FUTURE BIOMARKER
AND DRUG DEVELOPMENT.
SO WE HAVE A LARGE PORTFOLIO OF
TISSUES AND ORGANOIDS SO WE CAN
PUT INTO THESE CHIPS.
WHAT WE NOW CALL THE BODY IN A
CHIP.
AND BASICALLY A LOT OF
PUBLICATIONS THAT HAVE COME OUT
OF OUR TEAM HERE AT THE WAKE
FOREST INSTITUTE FOR
REGENERATIVE MEDICINE, JUST
REALLY A TERRIFIC TEAM.
AND I'D LIKE TO THANK YOU ALL
FOR YOUR ATTENTION AND THANK YOU
AGAIN FOR HAVING ME.
>> THANKS SO MUCH, TONY, FOR THE
VERY NICE OVERVIEW.
THINK THE BREAK WAS AT 2:40.
I THINK IF ALL THE SPEAKERS CAN
STAY FOR THE QUESTION AND ANSWER
SESSION, WHICH IS GOING TO BE, I
BELIEVE, AT 3:40, I THINK I
WOULD RATHER TAKE THE BREAK NOW
UNTIL 3:00, AND THEN WE'LL
RESUME THE SESSION AT 3:00 AND
THEN HAVE THE QUESTION AND
ANSWER SESSION AT 3:40.
IS THAT OKAY WITH EVERYONE?
SARINE, ARE YOU OKAY WITH THAT?
>> THAT'S PERFECT, IF EVERYONE
IS OKAY WITH THAT, WE SHOULD
STICK TO THE SCHEDULE.
>> ALL RIGHT.
THANK YOU, ALL THE SPEAKERS, AND
WE'LL RESUME AT 3:00.
>> 3:00.
>> SEE YOU IN A BIT.
AND KEEP ADDING QUESTIONS TO THE
BOX.
WHILE YOU'RE ON BREAK.
THANK YOU FOR THE INTRODUCTION.
I'M JASON, POSTDOC FROM MIT.
TODAY I'M GOING TO REPRESENT MY
SUPERVISOR, ROGER KAMM, TO SHARE
VASCULOGENESIS AND ANGIOGENESIS
MODELS.
FOCUSING ON THE NEUROLOGICAL
DISEASE, CANCERS, AND ALSO LAST
PART OF MY TALK, I'M GOING TO
SHOW ROBUST STRATEGIES TO FORM
MICROVASCULAR NETWORKS,
HOPEFULLY IT WILL BE BENEFICIAL
FOR ANTIVIRAL DRUG DEVELOPMENT
AS WELL.
SO AS WE KNOW,
MICROPHYSIOLOGICAL SYSTEMS
ACTUALLY ATTRACT MORE AND MORE
ATTENTION RECENTLY BECAUSE OF
PARTIALLY TO RECALCULATE -- THE
ORGANIZE FORM AND FUNCTIONS
THROUGHOUT THE CELLS MODIFY
SCREENING DRUGS TO MIMIC DISEASE
MODELS, TISSUES, ORGANS ACTUALLY
FOCUSING ON THE CIRCULATION
SYSTEM AND AS WE ALL KNOW THAT
BLOOD OR NUTRIENTS IN CELLS
TRANSPORT THROUGH THE
CIRCULATION SYSTEM AND THEY ARE
VERY IMPORTANT FOR ALMOST ALL
THE OR ORGANS.
PARTICULARLY WE ARE INTERESTED
IN THE CAPILLARY, ENGINEERING OF
CAPILLARIES.
THEY HAVE THREE FEATURES.
ONE IS THEY ARE PERFUSABLE AS I
JUST MENTIONED, THE CELLS GET
TRANSPORTED THROUGH THE
CAPILLARIES.
THE SECOND FEATURE IS THEY HAVE
SMALL DIAMETER.
SOMETIMES THEY HAVE ONLY SINGLE
CELL CAN CUT THROUGH ONE
CAPILLARY, THE THIRD FEATURE IS
THEY ARE PERMEABLE, SO THEY CAN
TRANSPORT ACROSS THE ENDOTHELIUM
FROM THE LUMINAL SIDE TO THE
STROMAL SIDE.
SO TO MIMIC CAPILLARIES BY
PROFESSOR MIMMERY AND ALSO
PROFESSOR GRIFFIN, SO AT THE LAB
WE ACTUALLY USE MICRO -- TO
ENGINEER CAPILLARY-LIKE
STRUCTURES, SO -- DESIGNS, MOST
OF THEM SIMILAR TO THIS A --
THREE CHANNELS.
IN THE CENTER THIS IS A GEL
CHANNEL, LEFT SIDE THEY HAVE
MEDIA CHANNEL.
ENDOTHELIAL CELLS AND STROMAL
CELLS INTO THE -- IN HYDROGEL
PRECURSOR AND WE INJECT IT --
WE'RE GOING TO ADD THE MEDIA
INTO THE DEVICE SO -- SO BE SINK
GEL CELL ARE GOING TO MIGRATE,
PROLIFERATE, MAKE CONNECTIONS
AND EVENTUALLY THEY ARE GOING TO
FORM A NETWORK SO -- SHOWN IN
GREEN SO WE CALL IT
MICROVASCULAR NETWORKS AND ALL
THESE DEVICES HAVE LIKE
COMMERCIALLY AVAILABLE FROM AIM
BIOTECH.
SO OUR DEVICE TECHNIQUE ACTUALLY
PREPARED WITH ASSAYS, WE CAN
TAKE FLUORESCENT IMAGING OR USE
TEM TO SEE THE STRUCTURE.
WE CAN ALSO ATTRACT MORE
INFLAMMATION SUCH AS GENOMIC
RNA, DNA SEQUENCING, COMPARED
WITH OUR DEVICE, WE CAN SORT
CELLS, ALSO COLLECT MEDIA TO SEE
HOW THE PROTEINS ARE SECRETED BY
THE CELLS, DO SOME FUNCTIONAL
ANALYSIS SUCH AS MICRO
VASCULATURE -- TO SEE TUMOR
CELL -- ET CETERA.
SO I'M GOING TO SHOW ALL THOSE
ASSAYS AND APPLICATIONS USING
THE FOLLOWING EXAMPLES.
SO AS I MENTIONED THAT THE
CAPILLARIES, ONE OF THE FEATURES
IS -- IN OUR SYSTEM WE CAN
ACTUALLY DETECT THE
PERFUSABILITY BY ADDING
FLUORESCENT MOLECULES SUCH AS
DEX TRAN THROUGH THE MEDIA
CHANNEL.
ACTUALLY SEE THE TRANSPORT
THROUGH THE LUMEN AND THE
VASCULAR IMAGE -- WE CALL SUCH A
STRUCTURE AS THEY ARE PERFUSABLE
MICROVASCULAR NETWORKS.
ANOTHER FEATURE OF THE CAPILLARY
IS THEY ARE PERMEABLE, SO IF WE
PERFUSE SMALL MOLECULES -- VERY
SMALL AND THEN WE TOOK TWO
IMAGES AT DIFFERENT TIMES, WE
SEE ACTUALLY HOWL RAPID THE --
ACROSS THE ENDOTHELIUM GET INTO
THE STROMA, YOU SEE THE STROMA
REGIONS HAVE HIGHER FLUORESCENT
SIGNALS.
WE PERFUSE -- SO THE MOLECULES
CAN GET THROUGH THE TIGHT
JUNCTIONS, -- CAN SEE THERE ARE
NO SIGNIFICANT FLUORESCENT
SIGNALS ACROSS THE LUMEN TO THE
STROMAL SITE.
SO IF YOU QUANTIFY THOSE
FLUORESCENT SIGNALS, WE CAN
ACTUALLY MEASURE THE PERM
PERMEABILITY OF THE VESSELS.
SO WHAT I WANT TO SHOW HERE IS
THE GREEN ONES ARE THE ONES IN
VIVO USING OUR IN VITRO SYSTEM.
SO JUST TO -- THE IMPRESSION
THAT OUR MODEL IS REALLY -- HAS
SIMILAR PERMEABILITY COMPARED TO
THE ONES MEASURED IN VIVO.
COMPARED TO THE TRADITIONAL
2D -- METHOD TO MEASURE THE
PERMEABILITY, THEY HAVE MORE --
THEY ARE BETTER, THEY HAVE A
LOWER MOBILITY.
SO THOSE ARE FROM USING THOSE
FLUORESCENT SIGNAL BASED ON
PERMEABILITY, WE ALSO SEE SOME
STRUCTURE FROM OUR MUTUAL MODELS
VERY SIMILAR TO THE ONES IN VIVO
THAT YOU CAN SEE THOSE TIGHT
JUNCTIONS, VERY TINY CONNECTION
AND ALSO YOU CAN SEE THE
VESICLES DURING THE ACTIVE
TRANSPORTATION.
SO WE GET THE IMPRESSION INDEED
OUR -- MAY MAKE IN VIVO
CAPILLARY WELL.
NOT ONLY WE CAN PERFUSE THOSE
MOLECULES BUT PERFUSE OTHER
PARTICLES, SO AS FROM HERE, RED
AND WE CAN ALSO USE SIMILAR
STRATEGY TO SEE THE PERMEABILITY
OF NANOPARTICLES TO COMPARE
WHETHER THEY ARE MODIFIED OR
WITHOUT MODIFICATION, CAN
REGULATE THE STRATEGIES TO
IMPROVE THE DRUG -- SO NEXT I'M
GOING TO TALK ABOUT THE BBB
SYSTEM, BLOOD BRAIN BARRIER,
QUITE A UNIQUE STRUCTURE, THEY
HAVE VERY TIGHT JUNCTION AND
ALSO THEY HAVE VERY HIGH
SELECTIVITY, ALL THE MOLECULES
CAN CUT THROUGH THE ENDOTHELIUM
AND INTO THE BRAIN SO YOU DON'T
WANT EVERYTHING GETTING -- THE
BRAIN TO MESS UP ANYTHING, SO
THEY HAVE SOME UNIQUE TRANSPORT
CHANNELS THAT HELP THE MOLECULES
TRANSPORTATION.
TO MIMIC SUCH -- STRUCTURE, THE
LAB ACTUALLY DEVELOPED A MODEL
USING -- ONE IS LIKE A HUMAN
DERIVED ENDOTHELIAL CELLS PAIRED
WITH THE BRAIN PERICYTES AND
ASTROCYTES, SO TWO DAYS THEY
FORM THE MICROVASCULAR NETWORKS,
SO WHAT WE FOUND IS -- CULTURE
CONDITIONS, THEY HAVE GOOD --
COMPARED TO THE MOUSE BRAIN
MEASURED IN VIVO.
SO -- STROMAL CELLS, THE
PERMEABILITY -- THIS TELLS US
TO -- WE NEED ALL THOSE STROMAL
CELLS.
SO NOT ONLY WE TEST THE
FUNCTION, WE ALSO TEST THE
TRANSCRIPTOME RESULTS OF USING
OUR MODELS, SO WE COMPARE THE 2D
AND 3D USING HUMAN BRAIN --
ENDOTHELIAL CELLS PAIRED WITH
PERICYTE AS MENTIONED.
SO THE RESULT 3D BETTER THAN 2D
AND ALSO WHEN WE HAVE
TRI-CULTURE, WE HAVE THE
PERICYTES OUTSIDE, THEN
ENDOTHELIAL CELLS EXPRESS HIGHER
LEVELS OF THOSE TIGHT JUNCTION
PROTEINS ANDS ARE AND ALSOS
AS AN EXAMPLE THAT -- SYSTEM
MIMIC BETTER THE -- SYSTEM IN
VIVO.
SO NOW BESIDES PROFUSE MOLECULES
ACTUALLY PERFUSE TUMORS BETWEEN
CELLS AS WELL.
SO HERE ARE SOME EXAMPLES OF
PERFUSED TUMOR CELLS, THEY WERE
GREEN -- MICRO VASCULATURE, WE
CAN SEE HOW THE TUMOR CELLS
EXTRAVASATED.
ACTUALLY MAKE A HOLE FROM -- IN
THE VESSELS AND THEN IN THIS WAY
THEY'RE -- MULTIPLE TYPES OF
CELLS, TUMOR CELLS, NEUTROPHILS
TO SEE HOW THOSE TWO CELLS
INTERACT WITH EACH OTHER.
SO USING COMBINED THE BB SYSTEM
AND ALSO TUMOR CELL MODEL, WE
ALSO STUDIED -- THEY PREFER
TO -- FOUND METASTASIZE WHEN
COCULTURED WITH TUMOR AND
ASTROCYTES ALONE THEY CREATE
HIGHER LEVEL OF CCL2, AND THOSE
ATTRACT TUMOR CELLS EXPRESSING
CCR2 TO WE ACTUALLY USE
ANTI-CCL2 TO BLOCK THE SECRETION
AND ALSO -- FROM THE TUMOR SITE
AND -- WE CAN REDUCE
EXTRAVASATION OF THE TUMOR
CELLS.
SO THIS IS AN EXAMPLE OF HOW TO
USE OUR MODEL SYSTEM, OUR --
SYSTEM TO STUDY TUMORS IN
EXTRAVASATION.
WE ALSO DEVELOP SOME SOLID
TUMOR -- MODEL, SO SIMILAR LE WE
HAVE A THREE-CHANNEL DEVICE, THE
MISSION OF THE DEVICE IS
ACTUALLY LARGER SO IN THIS WAY,
WE CAN EMBED A LARGER TUMOR --
INTO
SPHEROID INTO THE DEVICE SO WE
CAN ACTUALLY TEST THE
PERMEABILITY OF THE VESSELS AT A
DIFFERENT DISTANCE FROM THE
TUMOR.
THESE ARE CLOSE, THESE ARE FAR
AWAY FROM IT.
WE CAN ALSO USE THIS MODEL TO
SEE -- TO TEST DRUG
TRANSPORTATION.
OR WE CAN ALSO USE THIS
VASCULARIZED TUMOR MODEL TO TEST
FOR KA.-T-CELL CAR-T-CELL.
WHITE COLOR, WE CAN ACTUALLY SEE
HOW THOSE T-CELLS INTERACT WITH
THE TUMOR CELLS, WHICH IS SHOWN
IN RED, AND EVENTUALLY THEY CAN
SCAN THE DEAD CELLS TO SEE
CAR-T-CELL KILLING -- T-CELL
RESPONSE.
SO OTHER DEVICES THAT WE
ACTUALLY STUDIED THE
ANGIOGENESIS OF THE BLOOD
VESSELS ACROSS THE POROUS
MEMBRANES.
SO THERE IS A -- ON THE LEFT
SIDE -- THE NETWORKS FOR
TREATING THE DEVICE WITH OTHER
CYTOKINES OF THOSE -- ON THE
LEFT SIDE, SO THOSE ENDOTHELIAL
CELLS -- ANGIOGENESIS THROUGH
THOSE POROUS MEMBRANES ACROSS --
SO IN THIS WAY JUST ONE EXAMPLE
TO SHOW YOU OTHER DEVICES FROM
THE LAB THAT CAN BE USED --
STUDIES.
WE ALSO HAVE ANGIOGENESIS USING
LYMPHATIC ENDOTHELIAL CELLS.
SO INSTEAD OF PUTTING THEM INTO
THE GEL CHANNEL AND THE MEDIA
CHANNEL ON THE LEFT AND ON THE
RIGHT SIDE WE'RE GOING TO ADD DI
FLOW WE CAN SEE HOW THOSE
LYMPHATIC ENDOTHELIAL CELLS
ANGIOGENESIS -- STIMULUS, THAT
MAY MAKE THIS RELEVANT FOR
INFECTING -- ANG GENESIS.
ANGIOGENESIS.
WE ALSO DEVELOPED THIS MICRO
PUMP WE CALL A MICROHEART, SO WE
CAN CONNECT TO THE DEVICE,
IMMUNE CELLS, TUMOR CELLS, TO
SEE HOW THEY RESPOND.
VASCULARIZED ALZHEIMER'S DISEASE
MODEL.
SO I'M GOING TO TALK ABOUT THOSE
TWO ROBUST -- TO FORM
MICROVASCULAR NETWORKS AND
ALSO -- SPHRAT -- STRATEGI.
SO FOR THE -- AND OTHER LABS
WHICH WE USE PRIMARY CELLS, AND
WE KNOW PRIMARY CELLS HAVE BATCH
TO BATCH VARIATIONS.
IN THESE STUDIES, THEY ACTUALLY
COMPARED DIFFERENT LUNG
FIBROBLAST, SAME HUMAN UMBILICAL
VEIN ENDOTHELIAL CELLS TO FORM
MICROVASCULAR NETWORKS.
-- THEY ARE TOTALLY DIFFERENT.
SO THIS INDEED TELLS US THOSE
PRIMARY CELLS SIGNIFICANT AND
EVEN THOUGH YOU FIND A GOOD PAIR
OF STROMAL CELLS AND IND THEEL
YAL CELLS, THEY STILL HAVE -- IN
THEM.
SO WE TOOK ADVANTAGE OF -- USING
HTERT -- INTO THE ENDOTHELIAL
CELLS, FIBROBLASTS TO MAKE THEM
SIMILAR TO A CELL LINE -- THEY
CAN FORM MICROVASCULAR NETWORK
SIMILAR TO THE PRIMARY CELLS IN
TERMS OF THEIR MORPHOLOGY, AND
SINCE THEY ARE IMMORTGAGIZED, WE
CAN PASSAGE THEM OVER 20 TIMES,
THEY ARE STILL GOOD FOR MICRO
MICROVASCULAR NETWORK FORMATION,
NOT ONLY WE TRIED THIS IN HUMAN
ALSO -- DERMAL ENDOTHELIAL CELLS
TO MIMIC THE SKIN MODEL, SO ALL
THIS TELLS YOU -- THIS IS --
CELLS ARE GOOD SOURCES FOR
MICROVASCULAR NETWORK FORMATION.
AND ALSO THEY ARE SIMILAR TO A
CELL LINE SO IF THEY ARE VERY
EASY TO -- CRISPR TECHNIQUE AND
ALSO YOU CAN -- SHOWN HERE TO
SEE HOW THOSE MICRO VASCULATURE
RESPONDS TO THE TUMOR CELLS WHEN
THEY PERFUSE THEM TO THE
VESSELS.
WE CAN SIMILARLY TEST THE TUMOR
CELL EXTRAVASATIONS, AND ALSO
USE THOSE -- CELLS FOR THE SOLID
TUMOR MODELS AS I JUST
INTRODUCED PREVIOUS.
SO ANOTHER CHALLENGE WHEN WE TRY
TO GENERATE MICRO VASCULATURE
IS -- OUR LAB AND OTHER
PUBLISHED DATA FROM OTHER LABS
THAT MOST OF THE PERFUSIBLE
NETWORK, THEY TEND TO HAVE
LARGER DIAMETER VESSELS.
SO BUT COMPARED TO THE ONES IN
VIVO, THOSE VESSELS -- ACTUALLY
THEY ARE VERY SMALL.
-- AS SHOWN HERE THEY CAN BE --
BUT HOW TO -- ENGINEERING THOSE
SMALL VESSELS THEY ARE
PERFUSABLE IS CHALLENGING.
SO FROM ALL THE STUDY AND
ALSO -- STUDIES FROM OTHERS WE
FOUND THAT MANIPULATING THE
ENDOTHELIAL CELL CONCENTRATION
WE CAN ACTUALLY GET DIFFERENT
MORPHOLOGY OF THE VESSELS.
SO FOR EXAMPLE IF WE USE A
HIGHER CONCENTRATION OF
ENDOTHELIAL CELLS -- THEY ARE
PERFUSIBLE.
BUT IF WE USE A LOWER -- THEY
ACTUALLY CAN FORM THE VESSELS
VERY SMALL.
MOST PERFUSABLE -- THEY ARE
PERFUSE ABLE BECAUSE THEY HAVE
LARGE DIAMETER OPENINGS
CONNECTING TO THE MEDIA CHANNEL
TO ALLOW DEXTRAN TO FLOW THROUGH
THE NETWORKS.
THESE OPENINGS, THEY ARE CLOSED,
SO HOW ABOUT WE COMBINE THESE
TWO CONDITIONS SO WE CAN FORM A
LARGER DIAMETER VESSELS AT THE
REGION BETWEEN THE MICRO -- TO
COME BACK TO THE MEDIA CHANNEL
BUT AT THE CENTER WE HAVE VERY
SMALL, TINY VESSELS.
SO WE DEVELOPED -- FIRST WE SEE
THE HIGHER DENSITY OF
ENDOTHELIAL CELLS INTO THE
DEVICE AND HAS CAREFULLY
ASPIRATED THE -- BECAUSE OF THE
SURFACE TENSION PROVIDED BY THE
MICRO POST, THOSE HIGHER DENSITY
OF THE ENDOTHELIAL CELLS -- IN
THIS REGION AND THEY ARE GOING
TO FORM A LARGER DIAMETER VESSEL
OPENINGS.
LOWER -- WITH FIBROBLASTS INTO
THE CHANNEL AGAIN, SO IN THE
CENTER THEY HAVE ENDOTHELIAL
CELL -- THEY'RE GOING TO FORM A
VERY SMALL DIAMETER VESSELS, SO
IN THIS WAY, WE CAN GENERATE
MICRO VASCULATURE THAT HAS VERY
TINY SMALL DIAMETERS IN THE
CENTER CONNECTING TO A LARGER
OPENINGS ON THE SIDE.
AND I JUST SHOW VERY CROWDED
IMAGE HERE, THIS IS ACTUALLY --
IN MY DEVICE AND THOSE IMAGES
ARE -- AND ALL OF THOSE --
PERFUSIBLE AND WITH LARGER
OPENINGS ON THE SIDE AND SMALLER
VESSELS IN THE CENTER.
SO JUST TO GIVE YOU THE
IMPRESSION THESE TWO METHODS
REALLY ROBUST AND HOPEFULLY CAN
BE CONVINCING FOR THE COMMUNITY.
THIS TWO EPITHELIUM METHOD --
PATIENT DERIVED CELLS ORDER MALL
FIBROBLASTS TO MAKE THE SKIN
MODEL OR BRAIN -- OR USING --
ENDOTHELIAL CELLS.
SO AS A SUMMARY, -- ALSO I SHOW
USING MODELS COMPARED WITH OTHER
TECHNIQUES AND TO SEE HOW IT CAN
BE USEFUL -- NEUROLOGICAL
DISEASE OR CANCER STUDY, LAST I
SHOWED TWO STRATEGIES WHY TO
USE -- AS CELL SOURCES AND THEN
INTRODUCE THIS TWO-STEP METHOD
TO GENERATE THE VERY SMALL
VESSELS IN THE CENTER AND
THEN --.
LAST I WOULD LIKE TO THANK MY
SUPERVISORS, COLLABORATORS,
FUNDINGS.
AND THANK YOU VERY MUCH FOR YOUR
ATTENTION.
>> THANKS SO MUCH, JASON.
THIS WAS BEAUTIFUL WORK.
THANKS SO MUCH FOR SHARING.
WE'RE GOING TO KEEP GOING, AND
AGAIN IF YOU HAVE ANY QUESTIONS,
POP THEM IN THE Q & A BOX.
LAST SPEAKER, NOT THE LEAST, IT
IS MY PLEASURE TO INTRODUCE
DR. JEFF BORNENSTEIN, GROUP
LEADER FOR SYNTHETIC BIOLOGIES
AND BIOINSTRUMENTATION AND
LABORATORY FELLOW AT DRAPER IN
CAMBRIDGE WHERE HE LEADS
PROGRAMS IN ORGAN AND DISEASE
MODELS FOR DRUG DISCOVERY AND
SAFETY TESTING, ORGAN ASSIST
DEVICES AND DRUG DELIVERY
SYSTEMS.
JEFF IS MEMBER OF THE NATIONAL
ACADEMY OF INVENTORS AND HE'S
BEEN DOING A LOT OF WORK WITH
APPLICATIONS OF MICROSYSTEMS AND
MICRO FABRICATION TECHNOLOGIES
TOWARDS MEDICINE.
JEFF, THE FLOOR IS YOURS.
>> THANK YOU, MARC.
IS THAT SHARING PROPERLY?
>> YES.
>> GREAT.
THANK YOU SO MUCH, MARC, FOR THE
KIND INTRODUCTION, AND THANK
YOU, SARINE AND THE ORGANIZERS.
IT'S A PLEASURE TO BE HERE AS
PART OF THIS GREAT PANEL TODAY.
I'D LIKE TO TALK TO YOU ABOUT
OUR WORK IN DRAPER IN ORGAN ON
CHIP PLATFORMS TOWARDS THREE
DIMENSIONS, REALLY.
ONE OF THEM AS HAS BEEN TALKED
ABOUT EARLIER IS THE HIGHER
THROUGHPUT ASPECT.
ANOTHER IS WORKING IN HIGH
CONTAINMENT, PARTICULARLY ACUTE
NEED AND SITUATION WITH REGARD
TO COVID-19 AND MANY OF THE
OTHER HIGH PRIORITY PATHOGENS
THAT THE GOVERNMENT IS
INTERESTED IN IN PURSUING
MEDICAL COUNTERMEASURES FOR,
THEN REALLY ANOTHER EXCITING
DIMENSION, AND THAT'S
PERSONALIZED MEDICINE, WHERE WE
CAN OBTAIN CELLS FROM PATIENT
POPULATIONS AND UNDERSTAND HOW
THESE INFECTIOUS DISEASES AND
POTENTIAL TREATMENTS WORK IN
PARTICULAR PATIENT DEMOGRAPHICS
AND POPULATIONS.
SO WITH THAT, JUST A LITTLE BIT
OF BACKGROUND ON THE PREDICT
96 TECHNOLOGY.
AND SO PREDICT 96 IS REALLY
BUILT ON A PLATFORM, ON A BACK
PLANE OF A STANDARD MICRO MIR
PLATE.
THERE ARE FOUR WELLS FOR EACH
DEVICE.
SO ESSENTIALLY 96 DEVICE SYSTEM
BUILT ON TO THIS 3D PLATE.
THERE'S AN APICAL CHANNEL AND A
BASAL CHANNEL WITH AN
OVERLAPPING REGION THAT'S
SEPARATED BY A SEMI POROUS
MEMBRANE, AND THERE IS FLOW
THROUGH THE APICAL CHANNEL AND
THROUGH THE BASAL CHANNEL AND
CELLS CAN BE CULTURED ON ONE OR
BOTH SIDES OF THIS MEMBRANE.
SO THERE'S FOUR PORTS, TWO
INLETS, TWO OUTLETS, ARRANGED ON
THE 96 DEVICE SYSTEM.
WE CAN EITHER WORK WITH CULTURED
TISSUES AS SHOWN HERE WITH
MATRIX MATERIALS ON THE MEMBRANE
AND VARIOUS CELL TYPES, AS MANY
AS FOUR OR FIVE CELL TYPES, OR
WE CAN WORK IN OTHER
CONFIGURATIONS WITH ORGANOIDS OR
TUMORS THAT'S BEEN DISCUSSED
PREVIOUSLY.
I'D LIKE TO TOUCH ON SOME OF THE
POINTS THAT THIS PANEL HAS
TALKED ABOUT THROUGHOUT THE
COURSE OF THE DAY, AND REALLY
ONE OF THOSE IS SCALE, AND I
THINK THAT WAS AN IMPORTANT
POINT MADE EARLIER.
SO THE DESIRE HERE IS TO BE ABLE
TO STUDY MANY CONDITIONS AND
LOOK AT STATISTICAL REPLICATES
ON A SINGLE PLATE TO TRY TO
REDUCE SOME OF THE VARIABILITY
AND INCREASE THE THROUGHPUT OF
THESE SYSTEMS, AND SO THAT'S THE
GOAL HERE, AND I THINK ANOTHER
IS TO MAINTAIN THE FIDELITY OF
THE SYSTEM AND WE DO THAT WITH
SENSING AND FLUIDICS INTEGRATED
INTO THESE PLATES.
SO THERE'S A CLAMSHELL HERE THAT
PROVIDES A PUMPING TO EACH OF
THESE 96 DEVICES SO THE APICAL
CHAMBER AND THE BASAL CHAMBER OF
EACH OF THE 96 DEVICES IS
ADDRESSED SEPARATELY AND
INDEPENDENTLY BY THESE PUMPS
THAT ARE INTRODUCED FROM AN
OVERLYING CLAM SHELL STRUCTURE,
AND THEN AS WE'LL SHOW ON THE
NEXT SLIDE, THERE ARE READOUTS
HERE.
AND I THINK THE KEY REALLY WITH
THE THROUGHPUT IS NOT ONLY JUST
HOW MANY CONDITIONS ON A PLATE,
BUT HOW MANY PLATES CAN FIT INTO
AN INCUBATOR AND SO WE CAN SEE
WE'RE UPWARDS OF -- ONE OF THE
KEYS TO THE TECHNOLOGY IS TO BE
ABLE TO USE IT APPLYING STANDARD
TOOLS AND STRUCTURES AND IN
TALKING WITH THE PHARMACEUTICAL
LABORATORIES, THINK THE ISSUE OF
MAINTAINING THE FIDELITY OF
THESE MODELS WHILE ACHIEVING
HIGH-THROUGHPUT IS REALLY
CRITICAL.
ONE OF THE OTHER POINTS WE HEARD
EARLIER WAS THAT SOME OF THE
IMMORTALIZED SUS TEMS ARE SYSE
LACKING -- I THINK THE OTHER
SIDE OF THAT CHALLENGE, THOUGH,
IS THAT SYSTEMS LIKE VERO
E6 PRESENT KIND OF A CHALLENGE
FROM THE ORGAN ON A CHIP
STANDPOINT BECAUSE OF THEIR
ROBUSTNESS AND STATISTICAL POWER
AND THROUGHPUT, EVEN THOUGH THEY
LACK IF DEALT AND FIDELIE
POWER IN SOME CASES.
SO I THINK THE CHALLENGE IS TO
ACHIEVE HIGHER FIDELITY BUT TO
MAINTAIN THE THROUGHPUT,
USABILITY AND TO SUPPORT THE
WORKFLOW IN THE PHARMACEUTICAL
INDUSTRY WITH AN ORGAN ON A CHIP
SYSTEM.
SO YOU CAN SEE WE BUILT IN NOT
ONLY THE PUMPS BUT THE
INTEGRATED BARRIER FUNCTION AND
CURRENTLY WITH BARDA WORKING ON
CHEMICAL SENATOR INTEGRATION AS
WELL.
ONE OF SENSORS
WELL.
ONE OF THE PROBLEMS WITH
TRANSWELL SYSTEMS IS THAT THE
MEMBRANE IS TYPICALLY NOT FLAT
AND THE SYSTEMS MUST BE REMOVED
MECHANICALLY IN ORDER TO DO
BIRESOLUTION, HERE WE HAVE MUCH
SMALLER MEDIA VOLUME, LESS USE
OF REAGENTS IN DRUGS AND
COMPATIBILITY WITH IN SITU HIGH
CONTENT IMAGING.
SO ALL OF THOSE FEATURES COME
TOGETHER IN THE
PREDICT96 SYSTEM.
IF WE THINK ABOUT THE DIFFERENT
ORGAN MODEL, WE'VE WORKED IN
MANY DIFFERENT ORGAN MODELS AS
MANY OF THE OTHER SPEAKERS HAVE
TALKED ABOUT TODAY, THE BLOOD
BRAIN BARRIER WORK WE'RE DOING,
WE'RE WORKING AND HAVE WORKED
WITH PHARMACEUTICAL COMPANIES ON
MANY OF THESE MODEL, THE
LIVER/KIDNEY VASCULAR SYSTEM,
THE ONE WE'LL FOCUS ON TODAY IS
THE AIRWAY FOR THE RESPIRATORY
DISEASE APPLICATION.
MANY ARE PUBLICLY ACKNOWLEDGED
COLLABORATIONS WITH COMPANIES
LIKE COLGATE-PAL MO LIVE,
BRISTOL-MYERS SQUIBB.
SOME ARE NOT PUBLICLY
ACKNOWLEDGED BUT MANY OF THE TOP
PHARMA COMPANIES.
THEN THE ONE WE'LL TALK A LITTLE
MORE ABOUT HERE IS THE PROGRAM
LOOKING AT INFLUENZA, LOOKING AT
MEDICAL COUNTERMEASURES FROM THE
EMERGING FIELD OF PROGRAMMABLE
GENE MODULATORS.
THAT'S A PROGRAM WE'VE BEEN
INVOLVED WITH THAT'S HELPED
SUPPORT THE DEVELOPMENT OF THE
AIRWAY MODEL.
SO HERE'S A CLOSER LOOK AT THAT
AIRWAY MODEL, AND AGAIN, IT IS
THIS PREDICT 96 PLATFORM WITH 9L
PLANE.
THE ONLY DIFFERENCE IS THE
APICAL CHAMBER DOES NOT HAVE
PUMPING OR FLUID.
IT'S AT AN AIR-LIQUID INTERFACE
SO WE'RE PUMPING THE BASAL
CHAMBER WHICH SUPPORTS EXCELLENT
TISSUE DEVELOPMENT OVER THE
COURSE OF ROUGHLY 21 DAYS WITH
HIGH BARRIER FUNCTION, MUCOUS
FORMATION, CILIARY BEADING AND
MANY OF BEATING, MANY
OF THE CELL TYPES PRESENT THAT
ARE CRITICAL, GOBLET CELLS AND
OTHER CELLS, THE CLUB CELLS, SO
WE FORMED THIS TISSUE, THIS IS A
SIDE VIEW OF IT, IN A SCHEMATIC
FORM AND THEN A HIGH RESOLUTION
IMAGING AND WE CAN SEE THE
PSEUDOSTRATIFIED EPITHELIUM
THAT'S ESTABLISHED IN THIS
SYSTEM.
SO THIS IS PUBLISHED IN
SCIENTIFIC REPORTS, THE LEADER
OF THIS PROGRAM IS ASHLEY GARD,
WHO I WOULD LIKE TO ACKNOWLEDGE
HERE FROM OUR DRAPER TEAM.
ONE OF THE THINGS I MENTIONED AT
THE BEGINNING IS GETTING CELLS
FROM PATIENT DEMOGRAPHICS.
SO AN EXCITING ASPECT HERE THAT
WE'VE BEEN PURSUING FIRST WITH
DARPA FUNDING AND ALSO WITH OUR
SARS-COV-2 RESEARCH IS OBTAINING
OF CELLS FROM LIVING DONORS.
THIS IS WORK IN COLLABORATION
WITH THE MASSACHUSETTS GENERAL
HOSPITAL, DR. BEN MEDOFF AND DRY
CRITICAL CARE UNIT, AND THEY DO
THESE BRONCHOSCOPIES WHERE THEY
BRUSH AND THEN COLLECT CELLS
FROM THE AIRWAY, VARIOUS REGIONS
OF THE AIRWAY.
THEY DO SORTING AND THEN THEY
PROVIDE US WITH THESE
POPULATIONS AND WE CAN EXPAND
THEM AND DEVELOP THEM INTO THE
TISSUES.
AND AS I'LL TALK ABOUT LATER, WE
LOOK AT THE VARIOUS DEMOGRAPHIC
POPULATIONS AND HOW THEY WILL
INFLUENCE THE RESULTS THAT WE
OBTAIN WITH THESE INFECTIOUS
DISEASE MODELS.
AND WE GET SOME BEAUTIFUL TISSUE
FORMATION IN THESE SYSTEMS.
ONE OF THE INTERESTING THINGS TO
LOOK AT IS IN PLATE AFTER PLATE,
WE SEE VIVID DIFFERENCES BETWEEN
DIFFERENT DONORS, LIVING DONORS
BASED ON VARIOUS
CHARACTERISTICS.
THOSE DIFFERENCES ARE IN THE
PERCENT OF SILL CILIATED CELE
MUCOCILIARY BEATING AND MANY OF
THE OTHER ASPECTS OF THE
TISSUES, THE BARRIER QUALITY
THAT MAY BE INFLUENCED BY
BACKGROUND, GENDER, ETHNIC
BACKGROUND, IN PARTICULAR
COMORBIDITIES IS ONE OF THE MOST
INTERESTING ASPECTS HERE, CAN WE
UNDERSTAND WHAT DIABETES OR
ASTHMA, CHRONIC LUNG DISEASES,
WHAT INFLUENCE THAT MAY HAVE ON
THE PROGRESSION OF AN INFECTION
SUCH AS SARS-COV-2.
SO THAT IS ANOARL THING THAT
WE'RE LOOKING AT VERY
RIGOROUSLY.
SO I MENTIONED THE DARPA PREPARE
PROGRAM.
THAT'S AN INFLUENZA-FOCUSED
PROGRAM, AND WE'VE EVALUATED
MANY OF THE KEY INFLUENZA
STRAINS THAT THE PANDEMIC
STRAINS, HONG KONG 68 AND H1N1.
THIS IS PCR DATA AND WE CAN SEE
SEVERAL LOGS OF APPLICATION OVER
48 HOURS, AND WE CAN KNOCK DOWN
THAT VIRAL REPLICATION WITH A
DOSE-RESPONSE CURVE WITH THE
TAMIFLU AND WE CAN DO THAT WITH
DIFFERENT DONORS AND WE SEE
DIFFERENT RESULTS FOR THE
DIFFERENT DONORS.
SO COVID-19 HAS BEEN OUR MAIN
FOCUS RECENTLY.
I'D LIKE TO SPEND THE REST OF
THE TALK ON THAT ASPECT.
AND CERTAINLY THIS "NEW YORK
TIMES" ARTICLE ARTICULATES THE
COVID-19 VACCINE DEVELOPMENT,
THAT WAS DISCUSSED EARLIER AS
WELL AT THE BEGINNING OF THE
DAY, HAS BEEN A VERY POWERFUL
AND COMPELLING EXAMPLE OF HOW
THESE TECHNOLOGIES CAN COME
TOGETHER QUICKLY WITH SAFE AND
EFFICACIOUS VACCINES.
HOWEVER, I THINK A LOT OF PEOPLE
IN THE MICROPHYSIOLOGICAL
SYSTEMS DMEUNT, COMMUNITY, G
ABOUT COVID-19 THERAPEUTICS, ARE
SEEING AN OPPORTUNITY TO ADDRESS
ESSENTIALLY A GAP, AND THAT GAP
AS MANY OF THE SPEAKERS, ALL OF
THE SPEAKERS HAVE TALKED ABOUT,
IS IN THE PRE-CLINICAL
DEVELOPMENT.
IT'S IN THE LIMITATIONS OF
ANIMAL MODELS, IT'S IN THE
LIMITATIONS OF THESE SIMPLIFIED
IMMORTGAGIZED CELL CULTURE
SYSTEMS AND MICROPHYSIOLOGICAL
SYSTEMS HAVE THIS TREMENDOUS
OPPORTUNITY TO IMPACT AN AREA
LIKE THE SARS-COV-2 PANDEMIC AND
HOW ONE MIGHT IDENTIFY
THERAPEUTICS THAT ARE HIGHLY
EFFECTIVE FROM THESE SYSTEMS,
MUCH MORE RAPIDLY, INEXPENSIVELY
AND MUCH MORE ACCURATELY THAN
EXISTING PRE-CLINICAL MODELS.
SO WITH THAT, THAT'S ANOTHER
AXIS OF WHAT WE'VE TALKING ABOUT
HERE.
WE'VE TALKED ABOUT THE
HIGH-THROUGHPUT, WE'VE TALKED
ABOUT THE PERSONALIZED MEDICINE.
THE OTHER KEY ASPECT IS THE HIGH
CONTAINMENT OPERATION, AND I
THINK THEY ALL GO TOGETHER
ESSENTIALLY, AND ONE OF THE
THINGS WE'VE SEEN IS THAT MANY
OF THE EFFORTS WITH ORGANS ON
CHIPS, SOME OF THE THOSE NOTABLE
EFFORTS, HAS BEEN LIMITED TO
PSEUDOVIRUS AND VIRAL ENTRY, AND
WE KNOW THAT IN THAT CASE, THE
DYNAMIC RANGE IS VERY LIMITED,
AND MANY OF THE MECHANISMS OF
REPLICATION ARE NOT REFLECTED BY
JUST LOOKING AT A PSEUDOVIRUS,
AND SO OUR AIM EARLY ON IN THE
PANDEMIC WAS TO ESTABLISH A
CAPABILITY IN A BSL-3 LABORATORY
TO LOOK AT THE VIRUS AND ITS
REPLICATION, BOTH ACROSS STRAINS
BUT THEN WHAT HAPPENS WHEN WE
INTRODUCE THERAPEUTIC
CANDIDATES.
SO WE'VE LEASED SPACE AT THE
TUFTS UNIVERSITY AND BIOSAFETY
LABORATORY WEST OF HERE.
THESE ARE PEOPLE WITH DECADES OF
EXPERIENCE IN BSL-3 OPERATIONS.
IT WASN'T IN SARS-COV-2
OBVIOUSLY EARLY IN THE PANDEMIC,
IT WAS IN TUBERCULOSIS AND
RABIES AND OTHER
BSL-3 PATHOGENS, BUT THIS
EXPERTISE CAME TOGETHER WITH THE
TECHNOLOGY, WORKING TOGETHER IN
THIS FACILITY.
A LOT OF THE WORK HAS BEEN DONE
ON THE WASHINGTON STRAIN BUT
WE'RE ACQUIRING DELTA AND
OMICRON AS WELL, AND YOU CAN SEE
HERE THE LIST OF INFECTIONS
WE'VE BEEN WORKING WITH IN THE
BSL-2 AND BSL-3 LABORATORIES.
SO WITH THAT, WE WERE ABLE TO
DEMONSTRATE ROBUST VIRAL
REPLICATION OF SARS-COV-2 IN OUR
PREDICT 96 SYSTEM.
WE KNOW IT'S BEEN CHALLENGING,
WE'VE SEEN DISCUSSIONS IN THE
FIELD ABOUT HOW THIS IS
EXTREMELY CHALLENGING AND SOME
OF THOSE CHALLENGES MAY BE
ASSOCIATED WITH THE TRANS WELL
SYSTEMS OR WITH OTHER SYSTEMS,
THEY MAY BE ASSOCIATED WITH
OTHER LIMITATIONS IN THE
PROCESSES, BUT HERE WE'VE BEEN
ABLE WITH MORE THAN 20 OF THESE
PREDICT 96 PLATES TO DEMONSTRATE
ROBUST VIRAL REPLICATION, WE SEE
VERY LOW INTRA-PLATE VARIABILITY
SIX DAYS WHEN WE RE-INTRODUCE
THE INFECTION AT DAY ZERO, THIS
IS WHEN THE TISSUE HAS BEEN FULL
LEE DEVELOPED AND THEN WE HAVE
READOUTS.
IT ALSO GIVES US OPTIONS IN
TERMS OF THE WORKFLOW IF WE ARE
LOOKING AT LARGER PANELS OF
COMPOUNDS THAT ARE CANDIDATES
FOR EVALUATION AS AN ANTIVIRAL.
SO PCR IS KIND OF THE GOLD
STANDARD, BUT A LOT OF PLAQUE
ASSAYS, TCIB50 IS ATTRACTIVE
BECAUSE IT'S VERY
HIGH-THROUGHPUT.
WE CAN ALSO DO LEUM LUMINEX R
ASSAYS.
CERTAINLY RNA SEQ HAS BEEN DONE
ON A CASE-BY-CASE BASIS
PARTICULARLY IN THE DARPA
PROGRAM.
SO JUST GOING BACK TO THE
PRECISION MEDICINE ASPECT.
WE CAN SEE HERE'S SARS-COV-2
APPLICATION.
ACROSS MULTIPLE DONOR, WE CAN
LOOK AT THE CHARACTERISTICS OF
THESE DONORS.
THEIR GENDER, THEIR RACE, THEIR
AGE.
CAUSE OF DEATH IS LISTED HERE IN
THE CASE OF THESE CADAVER
SOURCES.
WE ALSO HAVE A SIMILAR PANEL,
I'M NOT SHOWING IT HERE, OF
LIVING DONORS FROM MASSACHUSETTS
GENERAL HOSPITAL, WHERE THESE
ARE OBTAINED FROM THE RESEARCH
BRONCHOSCOPIES, AND THEN THE
DOUBLING TIME AND OTHER
FEATURES.
THEY'RE JUST NOW GETTING INTO
THE THERAPEUTIC SCREENING.
SO WE CAN SEE ON A SINGLE
PREDICT 96 PLATE, WE CAN LOOK AT
AS MANY AS SIX OR MORE COMPOUNDS
AND HAVE REPLICATES AT VARIOUS
CONDITIONS, AND THESE CONDITIONS
CAN BE MULTIPLICITY OF
INFLATION, THEY CAN BE THE KOAS.
DOSE.
SO WHAT WE'VE DONE IS WE'VE
TAKEN MANY OF THE SORT OF
PREDOMINANT COMPOUNDS THAT --
TREATMENTS THAT ARE BEING USED,
THE APPROVED COMPOUNDS HERE IN
THE UPPER ROW AND ASK OURSELVES
THE QUESTION, THIS IS REALLY A
LITMUS TEST FOR ORGAN ON CHIP
SYSTEMS THAT WE'VE BEEN
UNDERTAKING HERE, AND THAT IS IF
WE INTRODUCE THE APPROVED
TREATMENTS FOR SARS-COV-2, CAN
WE REPRODUCE ESSENTIALLY WHAT IS
SEEN CLINICALLY, AND THEN IF WE
LOOK AHEAD, COULD WE THEN HAVE
FREE
PREDICTED WITHOUT GOING THROUGH
SCREENS OR ANIMAL MODELS OR
EXTENSIVE CLINICAL STUDY, COULD
WE PREDICT WHICH DRUGS ARE GOING
TO WORK BEST.
AND SO WE'RE LOOKING HERE AT
PAXLOVID, REMDESIVIR,
MOLNUPIRAVIR AND SOME OTHER
COMPOUNDS THAT ARE ESSENTIALLY
BEING EVALUATED.
CALPEPTIN AND OTHER COMPOUNDS.
SO JUST GOING TO THIS SLIDE HOOR
HERE WHERE WE ESSENTIALLY LOOK
AT THE RESULTS, WE THOUGHT OF
THOSE IN TERMS OF THE FULL
REDUCTION IN THE VIRAL RNA, AND
HERE YOU CAN SEE THE CURVES BUT
THESE BAR CHARTS ARE A LITTLE
EASIER TO VISUALIZE.
WHAT WE'VE DONE IS WE'VE ORDERED
THESE COMPOUNDS IN TERMS OF OUR
PREDICTION BASED ON THE PREDICT
96 STUDIES, AND WE CAN SEE A
TREMENDOUS FULL REDUCTION,
ESPECIALLY AT THE HIGH DOSES FOR
THE MAIN COMPONENT OF PAXLOVID
BUT EVEN AT THE MID RANGE DOSE,
THE REMDESIVIR IS KIND OF AN
INTERMEDIATE CASE WHERE AT VERY
HIGH DOSE, WE SEE VERY LARGE
REDUCTION IN THE VIRAL COPY
NUMBER, AND THEN THE
MOLNUPIRAVIR IS EASILY THE
WEAKEST OF THE THREE, WHERE
WE'VE GOT MARGINAL REDUCTION
EVEN AT THE HIGH DOSE, AND
REALLY VIRTUALLY NO EFFECT AT
THE LOWER DOSES.
AND IF WE JUST TAKE A LOOK AT
CLINICAL RESULTS HERE AND LOOK
AT THE HOSPITALIZATION, WE KNOW
PAXLOVID IS HIGHLY EFFICACIOUS,
CLEARLY THAT'S THE TREATMENT OF
CHOICE REDUCING HOSPITALIZATION
BY 89%.
THE REMDESIVIR REDUCES
HOSPITALIZATION ALMOST BY THE
SAME AMOUNT BUT MANY CLINICAL
STUDIES SUGGEST IT'S NOT QUITE
AS EFFICACIOUS AT PAXLOVID.
IT LOOKS THAT WAY FROM OUR
PREDICTION.
AND THE MOLNUPIRAVIR IN PLATE
AFTER PLATE SHOWS A RELATIVELY
WEAK RESPONSE IN OUR SYSTEM, IN
OUR HANDS, BUT ALSO IN TERMS OF
THE CLINIC, REDUCING
HOSPITALIZATION ONLY MARGINALLY.
IN ADDITION, IT DOES APPEAR TO
HAVE SOME SAFETY CONCERNS, BUT I
THINK WHAT WE'RE ESSENTIALLY
SHOWING HERE IS THE ABILITY OF
THIS PREDICT 96 SYSTEM TO GAUGE
THE EFFECTIVENESS IN TERMS OF
THE CLINIC OF THESE VARIOUS
COMPOUNDS IN THE SARS-COV-2
VIRUS, WHICH IS A CHALLENGING
VIRUS TO WORK WITH, AND SO WE'RE
VERY EXCITED OBVIOUSLY BY
THIS -- BY THESE RESULTS AND
BELIEVE THAT THE LOGICAL NEXT
STEP IS TO BEGIN TO LOOK AT
LARGER PANELS, LOOK AT DIFFERENT
PATIENT POPULATIONS AND LOOK AT
EMERGING VARIANTS WHERE WE WON'T
KNOW HOW WELL THESE DRUGS WORK.
IN FACT, SOMEONE YESTERDAY WAS
ASKING ME, CAN WE LOOK AT THE
REBOUND EFFECT IN PAXLOVID, AND
THAT'S A FASCINATING QUESTION.
SO I THINK THAT WOULD BE A NEXT
STEP.
SO JUST IN QUICK SUMMARY HERE,
THERE'S A CRITICAL NEED FOR
THESE ORGAN ON CHIP MODELS WITH
HIGH PRIORITY PATHOGEN RESEARCH.
THE HIGH-THROUGHPUT IS KEY.
WE'VE BEEN ABLE TO DEMONSTRATE
ROBUST VIRAL REPLICATION ACROSS
MANY, MANY PLATES IN THE
BSL-3 LABORATORY AND DONE SO
WITH CELLS FROM HUMAN PRIMARY
DONORS, AND WE REALLY BELIEVE
THAT THIS MCM SCREENING IS A
ROBUST AND REPRODUCIBLE
CAPABILITY, AND WE'D LIKE TO
CONTINUE WITH THAT WITH LARGER
PANELS AS I TALKED ABOUT ACROSS
THE VARIOUS DOMAINS THAT WE'RE
INTERESTED IN LOOKING AT.
SO WITH THAT, I'D REICH LIKEK
EVERYONE, PARTICULARLY ASHLEY
GARD, THE PRINCIPAL INVESTIGATOR
FOR THIS WORK AND HER
OUTSTANDING TEAM AT DRAPER, OUR
COLLABORATORS AT MGH, THE
OUTSTANDING TEAM AT THE TUFTS,
OUR UMASS COLLABORATORS ON THE
PREPARE PROGRAM AND THE FUNDING
SOURCES, AND THANK YOU ALL.
>> THANK YOU, JEFF.
THIS IS AMAZING WORK THAT YOU
GUYS ARE DOING AND THE
PHARMACOLOGICAL RESULTS AND THE
HIGH-THROUGHPUT THAT YOU'RE
GETTING IN YOUR PLATFORM.
IT'S REALLY ENCOURAGING AND
REPRODUCIBILITY.
I THINK NOW WE ARE IN THE
QUESTION AND ANSWER TIME.
BEFORE WE GO INTO THE DISCUSSION
AND QUESTIONS, I WOULD LIKE TO
THANK THE SPEAKERS AGAIN FOR
TERRIFIC PRESENTATIONS, AND
REALLY IN MY MIND, I THINK WE
GOT A VERY NICE BROAD VIEW OF
THE DIFFERENT TISSUE MODELS,
ORGANOIDS, TISSUE ON A CHIP.
FOR THOSE OF YOU WHO ARE NEW IN
THIS WORLD OF 3D TISSUE MODEL,
YOU GOT A VERY NICE PERSPECTIVE
OF WHAT'S HAPPENING.
THE TECHNOLOGY IS ADVANCING AT A
VERY FAST PACE, AND I THINK --
ONE OF THE CHALLENGES, I THINK
JOHN MENTIONED AT THE BEGINNING,
IS HOW DO YOU MAKE THIS PROMISE
INTO A REALITY.
OF ALL THE SPEERK, I SPEAKY
TWO THIRDS ARE BIOENGINEERS,
BIOENGINEERING LABS.
ONE OTHER QUESTION IS HOW DO WE
GET THESE TECHNOLOGIES INTO MORE
BIOLOGY LABS SO THERE'S MORE
PEOPLE REALLY DOING SCIENTIFIC
VALIDATION AND PHYSIOLOGICAL
VALIDATIONS OF THESE MODELS.
SO THERE'S A BUNCH OF QUESTIONS.
I'M GOING TO GO THROUGH THE
QUESTIONS IN THE BOX, BUT
BEFORE, I WANTED TO START WITH
CHRISTINE MUMMERY.
BECAUSE IF I UNDERSTOOD
CORRECTLY, YOU'RE NOT IN A
TISSUE ENGINEERING LAB.
YOU'RE A STEM CELL LAB.
SO MY QUESTION TO YOU IS, WHEN
YOU GOT INTO THESE 3D TISSUE
MODELS IN TERMS OF ORGANOID,
MICRO TISSUES AND THEN YOU'RE
DOING SOME WORK WITH TISSUE
CHIPS NOW THAT YOU SHOW, WHAT
WERE THE CONSIDERATIONS, WHAT
WERE THE DRIVERS, WHAT WERE THE
DECISIONS YOU HAD TO MAKE WHEN
YOU STARTED WORKING WITH THESE
COMPLEX TISSUES?
>> WELL, SO I'M A PHYSICIST BY
TRAINING, BUT I ADOPTED STEM
CELL BIOLOGY VERY EARLY ON.
SO I TRY TO COMBINE
DEVELOPMENTAL BIOLOGY CONCEPTS,
OR WHAT DOES A CELL NEED, WITH
ENGINEERING CONCEPTS.
BECAUSE I'M IN A ENVIRONMENT
WHERE -- I JUST THOUGHT CHIPS
WERE SO COOL, BUT THEY'RE USING
THE WRONG CELLS.
SO I'M NOT A FAN OF CELLS LIKE
HUVEX, WHICH ARE REALLY SORT OF
OLD ENDOTHELIAL CELLS, THEY'RE
NOT REALLY REPRESENTATIVE OF
WHAT YOU FIND IN MOST OF THE
BODY.
AND I -- WE'RE USING THESE
FANTASTIC STEM CELL MODELS BUT
YOU JUST PUT THEM IN THE WRONG
SUBSTRATE.
SO WE GOT TOGETHER AND DECIDED
TO TRY AND COMBINE THESE THINGS.
NOW, MOST IMPORTANT WAS TO MAKE
THE BIOLOGY ROBUST, SO I THINK I
SHOWED EXAMPLES OF HOW WE DO QA
AND QC BEFORE WE PUT THINGS IN
CHIPS, BUT THEN YOU ASK WHAT
CHIPS DO YOU USE.
SO YOU WANT SOMETHING THAT'S
AVAILABLE, REPRODUCIBLE.
IF YOU GO TO A TECHNICAL
UNIVERSITY, THEY'LL GIVE YOU A
CHIP AND THERE ARE LIKE 10 OF
THEM, WHEN YOU SAID YOU WANT A
HUNDRED, THEY CHANGED THEIR
DESIGN.
SO THERE IS THIS BALANCE BETWEEN
INNOVATION AND REPRODUCIBILITY.
SO THOSE ARE THE MOST OF THE
QUESTIONS WE ADDRESS IN CHOOSING
OUR CHIPS.
SO WE PROBABLY HAVE FIVE OR SIX
DIFFERENT COMMERCIAL CHIPS WE
USE.
EACH ARE SUITED FOR DIFFERENT
PURPOSES.
SO THAT'S MOSTLY HOW THE
REASONING HAS GONE.
I THINK MOST EXCITING IS THAT WE
CAN SEE THINGS IN 3D UNDER FLOW
THAT WE NEVER SAW IN 2D.
AND IT'S PUTTING THINGS IN SOFT
SUBSTRATES HAS BEEN REALLY
EXTREMELY HELPFUL.
>> SOFFIT FOR PURPOSE, .
SIMON MENTIONED AT THE
BEGINNING, WHAT'S THE QUESTION
YOU'RE ASKING, AND REALLY VERY
CRITICALLY WHAT IS THE BEST
PLATFORM TO ANSWER THAT
QUESTION.
LIKE YOU SAID, IT MIGHT BE MANY
PLATFORMS, DIFFERENT THROUGH
PUT.
THANK YOU, CHRISTINE.
THERE'S A QUESTION IN THE CHAT
ABOUT FLOW, AND I'M GOING TO ASK
JEFF AND JASON MAYBE TO TALK A
LITTLE BIT ABOUT FLOW, I THINK
THE QUESTION IS ABOUT SOMEONE
MENTIONED LIKE SHEER STRESS AND
HOW DO YOU CONTROL -- STRESS
EFFECTS ON FISH EU CELL -- MEDIA
FLOW RATES ON EACH MODEL?
JEFF FIRST, THEN MAYBE JASON,
CAN YOU GUYS COMMENT ON THAT?
>> I CAN QUICKLY ANSWER AND THEN
JASON PROVIDE ON HIS.
SO FOR OUR SYSTEM, THE SHEER CAN
RANGE ANYWHERE FROM ZERO UP TO
ABOUT 10 OR MORE DINES PER
SQUARE COUNCILMEMBER.
I CENTIMETER.
WE HAVE A HIGH SHEER FORMAT THAT
WE CAN USE IF FOR INSTANCE WE'RE
STUDYING LONG COVID, I THINK
THAT WOULD BE A SITUATION WHERE
YOU'D WANT TO GO UP TO THAT 10
DINES PER SQUARE CENTIMETER.
MANY OF THE OTHER APPLICATIONS,
WE WORK IN A SORT OF LOW TO MID
RANGE SHEER THAT MAY BE
APPROPRIATE, MAYBE ONE DINE PER
SQUARE -- WHERE THERE ARE
SHEER-SENSITIVE CELLS BUT WE CAN
DIAL IT OR TUNE IT ESSENTIALLY
FOR THE APPROPRIATE APPLICATION.
>> SO FOR MINE, WE USE A SIMILAR
DINE ZERO FLOW RATE, SO WHAT I
WANT TO COMMENT IS THE DIFFERENT
PUMP SYSTEM DO YOU WANT TO USE
IF YOU'RE CIRCULATING -- REQUIRE
TO MAINTAIN THE CONTINUOUS FLOW,
AND THAT WAY WE WILL CHOOSE THE
SIMPLEST WAY, JUST TO PUT -- ON
PATH THEAND USING THE GRADIENT
OF THE DIFFERENT -- WE DID TO
DROP FLOW.
SO IN THIS WAY, ALTHOUGH -- THEY
ARE NOT STABLE, BUT IN A SHORT
TIME, THEY ARE ACTUALLY QUITE
STABLE AND IT'S EASY TO AVOID
ANY -- VOLUME BY CONNECTING THE
PUPS
PUMPS OR ANY -- MICRO FLUID
DEVICE ALWAYS KILLS EVERYTHING.
SO ANOTHER THING I WANT TO
MENTION, PREVIOUS PUBLICATION
ABOUT THE MICRO HEART.
ACTUALLY -- HE DEVELOPED A WAY
TO QUANTIFY WHAT WILL BE THE
FLOW RATE BY USING CONSOLE
SIMULATIONS MAYBE YOU WANT TO
USE THAT AS WELL.
>> TONY, ACTUALLY THAT WAS A
QUESTION FOR YOU AS WELL.
THE QUESTION WAS, WHAT IS THE
CORRECT FLOW OF CIRCULATORY
SYSTEM THROUGH DIFFERENT ORGANS
AND THEIR PROPER METABOLISM IN
KIDNEYS OR LIVER.
HOW DO YOU -- WHEN YOU MAKE
THESE ORGANS ON A CHIP, WHEN YOU
HAVE DIFFERENT ORGAN, HOW DO YOU
ADJUST THE FLOW AND THE OTHER
QUESTION THAT COMES UP IS WHAT
MEDIAS DO YOU USE WHEN YOU HAVE
THESE MULTIORGAN CHIPS?
TONY, CAN YOU ADDRESS THIS
QUESTION?
>> YES, ABSOLUTELY.
THANK YOU, MARC.
GREAT QUESTION ACTUALLY BECAUSE
THIS IS AN IMPORTANT FACT.
WHEN YOU'RE PUTTING THESE TISSUE
MODELS ON THE CHIP, YOU HAVE TO
ACCOUNT FOR A COUPLE OF THINGS.
THE FIRST ONE IS THE VOLUME OF
THE ORGAN ITSELF.
SO YOU KNOW THE LIVER HAS MUCH
MORE VOLUME THAN THE HEART.
THE LUNG HAS MORE VOLUME THAN
THE HEART AND LESS THAN THE
LIVER, SO YOU HAVE TO
ACCOMMODATE FOR THAT IN YOUR
CHIP SYSTEM SO THAT YOUR BODY IN
A CHIP REALLY DOES REPRESENT THE
TRUE VOLUME IN A HUMAN.
YOU DON'T WANT TO HAVE 10 TIMES
THE HEART THAN YOU DO OF THE
LIVER.
SO THAT'S THE FIRST THING.
THE SECOND THING IS IN TERMS OF
THE FLOW, THE FLOW IS ACTUALLY
CONSISTENT WITH A MICROSYSTEM OF
WHERE WE REDUCE THE MICRO
FLUIDICS AND MEASURE THE FLUID
WITHIN THE SYSTEM.
DEPENDING ON HOW WE CONSTRUCT
THAT SYSTEM, BECAUSE AS YOU
KNOW, WE HAVE BEEN PRINTING THEM
AND WE HAVE ALSO BEEN MAKING
MICROPHYSIOLOGICAL SYSTEMS WITH
WELLS AND OTHER SYSTEMS.
EACH ONE IS DIFFERENT, AND YOU
REALLY DO HAVE TO FIGURE OUT
WHAT THE CONVERSION FACTOR IS IN
TERMS OF THE FLOW THAT GOES TO
THE BODY, BECAUSE IF YOU REALLY
THINK ABOUT IT, YOUR OWN BLOOD
FLOW GOES TO YOUR ENTIRE BODY,
YOU KNOW, EVERY HOUR BASICALLY,
YOU KNOW, YOU HAVE YOUR ENTIRE
BLOOD GO THROUGH YOUR BODY, SO O
YOU HAVE TO ACCOUNT FOR THE SALE
THING, WHICH IS ABOUT 4 TO
5-LITERS OF BLOOD THROUGHOUT
YOUR SYSTEM.
THE OTHER THING IS THE ABILITY
OF THESE STRUCTURES TO INTERACT
WITH EACH OTHER AND HOW ARE YOU
GOING TO MEL SURE THAT FLOW IN
TERMS OF THE MEDIA.
THE QUESTION WAS ABOUT OF THE
MEDIA.
ACTUALLY THAT WAS ONE OF THE
FIRST DIRECTIVES OF THE INITIAL
DTRA PROGRAM.
I WAS HAPPY TO SEE CLINT, HE WAS
ONE OF THE PEOPLE THAT ASSIGNED
THAT INITIAL PROGRAM WITH DTRA,
AND THE GOAL OF THAT PROGRAM WAS
COMMON MEDIA.
AND THAT'S WHERE WE WERE ABLE TO
DO, IS TO CREATE A COMMON MEDIA
THAT IS ABLE TO -- THAT ALLOWS
THE SYSTEM TO BE IN A STEADY
STATE IN TERMS OF EVERYTHING
THAT'S NEEDED WITHIN THE SYSTEM
AND THE SYSTEM SELF REGULATES,
SO WHAT HAPPENS IS INITIALLY THE
MEDIA GOES THROUGH AND THE
SYSTEM ITSELF HELPS TO REGULATE
THE MEDIA SO YOU HAVE SOMETHING
THAT IS GOOD FOR ALL THE CELL
TYPES WITHIN THAT SYSTEM.
SO THIS WAS ACTUALLY ONE OF THE
PRIMARY DIRECTIVES OF THE
DEFENSE AGENCY SYSTEM AND IT'S
WORK VED WELL FOR US.
>> THANKS, TONY.
IT'S ACTUALLY -- KIND OF SPLIT
THE QUESTION INTO TWO QUESTIONS.
I'LL START WITH YOU, TONY,
BECAUSE THE QUESTION WAS TO YOU
DIRECTLY.
ARE YOU USING HUMAN PRIMARY
CELLS OR ARE YOU USING IPS
DERIVED CELLS?
WE'VE SEEN IN SOME OF THE TALKS
THAT PEOPLE USED BOTH IN THE
SAME TISSUE, PRIMARY CELLS AND
IPS DERIVED CELLS, SO CAN YOU
TELL US A LITTLE BIT ABOUT HOW
DO YOU CHOOSE THE CELLS THAT YOU
USE FOR A PARTICULAR SYSTEM?
>> ANOTHER GREAT QUESTION.
SO WE REALLY TRY TO USE NORMAL
PRIMARY HUMAN CELLS WHEREVER
POSSIBLE.
SO FOR THE LIVER STRUCTURES, YOU
SAW THOSE LIVER CELL TYPES TO
CREATE THE TISSUES -- THERE ARE
SOME INSTANCES WHERE YOU HAVE TO
USE THE IPS.
WE'D RATHER NOT, BUT SOMETIMES
YOU HAVE TO BECAUSE THERE'S NO
EQUIVALENT TO THAT.
AND SO WHEN THAT'S THE CASE,
THEN WE WILL USE THE IPS CELLS.
BUT CERTAINLY WE DO NOT USE ANY
CELL -- LIKE CHRISTINE MUMMERY
SAID, I ENJOYED YOUR TALK,
CHRISTINE, LIKE SHE SAID,
BASICALLY YOU DON'T WANT TO BE
USING CELL LINES, RIGHT, YOU
DON'T WANT TO BE USING HUBIK
CELLS, THEY'RE
NON-REPRESENTATIVE OF THE NORMAL
SYSTEM.
SO WHEN WE CREATE THESE
STRUCTURES, WE REALLY WANT TO
USE NORMAL PRIMARY HUMAN CELLS
AS MUCH AS POSSIBLE.
>> OR IF I MAY ADD, YOU HAVE TO
DO A HEAD TO HEAD COMPARISON
WITH YOUR IPS DERIVATIVES.
THAT'S HOW WE DISCOVERED, OF
COURSE, THAT THEY WERE IMMATURE
AND THAT PUTTING IN STROMA MAKES
THEM TO A CERTAIN EXTENT MATURE.
SO YOU CAN PEN CH MARK AGAIN THE
PRIMARY TISSUES.
PRIMARY TISSUES ARE SOMETIMES
SUBJECT TO STRESS, SO YOU GET A
LOT OF STRESS GENES AND YOU HAVE
TO TAKE CARE OF THAT.
BECAUSE IT'S NOT -- I MEAN, A
LUNG LAVAGE IS DIFFERENT BECAUSE
THAT'S NOT STRESS, BUT IF YOU
WANT TO ASSOCIATE KIDNEY OR SOME
OTHER TISSUES, THE STRESS GENES
GO ALL OVER THE PLACE.
SO DURING SINGLE CELL RNA SEQ,
THIS IS NOT NECESSARILY
INFORMATIVE ALL THE TIME.
SO I THINK THIS IS JUST USING A
BIT OF COMMON SENSE TO FIGURE
OUT WHAT'S THE BEST FOR YOUR
SYSTEM, WHAT'S YOUR QUESTION,
WHAT DO YOU WANT TO KNOW THE
ANSWER TO WILL GUIDE US.
SO IT'S CLEAR WE CAN, OF COURSE,
GET MANY, MANY TISSUES FROM
MANY, MANY PATIENTS IF WE USE
IPS, AND YOU HAVE THIS
POSSIBILITY OF ALMOST
NEUTRALIZING BATCH TO BATCH
DIFFERENCES.
SO THAT HAS AN UPSIDE.
THE DOWNSIDE, AS I SAID, RIDES
UP FRONT OF THE IMMATURITY OF
THE SYSTEM AND MANY AMOUNTS OF
MANIPULATION.
SO I THIL STIL STILL THINKS
BENEFITS TO HAVING IPS OR HUMAN
ORGANOIDS.
WHAT'S NOT BEING COMBINED YET,
AND WE'RE GOING TO BE DOING THIS
WITH PEOPLE WORKING WITH HANS
CLEVERS IS COMBINING THE
ORGANOIDS WITH IPS -- WHAT YOU
CANNOT DO WITH THE ADULT
ORGANOIDS IS MAKE STROMA.
YOU CAN'T MAKE THE IMMUNE SYSTEM
ISOGENICLY -- YOU CAN GET IT FOR
PATIENTS BUT YOU CAN'T DO IT --
AND FROM IPS, CAN YOU MAKE
FAIRLY GOOD MONOCYTES.
YOU CAN MAKE THE TISSUE
APPROPRIATE STROMA, BUT YOU
CAN'T GET MATURE EPITHELIAL
COMPONENT THE WAY YOU CAN GET
THAT OF ORGANOIDS.
SO WHAT YOU WANT TO KNOW, YOU
WANT IT AS SIMPLE AS POSSIBLE TO
ANSWER YOUR QUESTION.
AS COMPLEX AS NECESSARY TO MODEL
YOUR DISEASE OR DRUG RESPONSE.
>> CHRISTINE, TO FOLLOW UP IN
ONE OF THE TALKS, I FORGOT IF IT
WAS YOU OR SOMEONE ELSE, THE
WHOLE ASPECT OF CONTEXT, TISSUE
CONTEXT, RIGHT?
YOU START WITH AN IPS FROM A
SKIN FIBROBLAST AND THEN YOU'RE
TRYING TO MAKE A LONG EPITHELIAL
CELLS, ARE YOU LOSING THE SORT
OF TISSUE CONTEXT OR BY JUST
PUTTING OTHER CELLS, STROMA
CELLS, YOU WILL GAIN THAT TISSUE
CONTEXT THAT MIGHT DIFFERENTIATE
ALSO THE CELLS, ENDOTHELIAL
CELLS --
>> SO YES AND NO.
SO THE STROMAL CELLS LIKE THE
ENDOTHELIAL CELLS CAN GET FROM
IPS CELL USING A STANDARD
PROTOCOL ARE STILL PLASTIC.
IF YOU CO-CULTURE THEM IN AN
ORGANOID OR A PARTICULAR TISSUE
TYPE, THEY UPREGULATE
TRANSCRIPTION FACTORS OF THE
TISSUE THEY'RE IN.
SO THEY'RE PLASTIC ENOUGH TO
ADULT IDENTITY.
SO IN THE FIRST PLACE, WE DID IT
THE DIFFICULT WAY.
WE GOT THE PROGENITOR OF THE
ORGAN SO IN THIS CASE, IT WAS
THE CARDIAC PROGENITOR, WE MADE
A CARDIAC ENDOTHELIAL CELLS, AND
THEN WE LOOKED AT THE
TRANSCRIPTION PROFILE AND WE SEE
CARDIAC TRANSCRIPTION FACTORS IN
A BONA FIDE ENDOTHELIAL CELL.
THAT TELLS BUT TISSUE
SPECIFICITY AND THAT WAS AN
EXTRA SORT OF MESSAGE, DON'T USE
HUVEX, BECAUSE THEY'RE NOT TRUE
TISSUE-SPECIFIC CELLS.
ON THE OTHER HAND, IT TURNED OUT
THAT PLASTICITY MEANT WITH A
CERTAIN A COCULTURE, YOU COULD
RECAPITULATE SOME OF THE
TISSUE-SPECIFIC FEATURES AND THE
SAME IS TRUE OF FIBROBLASTS TO
SOME EXTENT.
SO IT DEPENDS WHAT YOU WANT TO
DO.
IF YOU JUST WANT SOMETHING THAT
GIVES YOU FLOW, THAT'S FINE.
IF YOU WANT TO DO SOMETHING
ABOUT -- LEARN SOMETHING ABOUT
THE BIOLOGY OF VASCULAR CELLS,
THE VASCULATURE YOU PUT IN, THEN
IT'S A COMPLETELY DIFFERENT
QUESTION.
>> IF I MAY ANSWER THAT, BECAUSE
YOU KNOW ONE OF THE STRATEGIES
THAT WE HAD TO USE AS I
MENTIONED IS REALLY USING THE
EXTRACELLULAR MATRIX IN THE
ORGANOIDS.
THE REASON FOR THAT IS THAT WHEN
YOU'RE LOOKING AT NORMAL TISSUE,
WHEN YOU'RE LOOKING AT THE
ORGAN, THE ORGAN AT THE END OF
THE DAY IS BASICALLY THE CELLS,
AND THE EXTRACELLULAR MATRIX.
THAT'S WHAT IT IS, RIGHT?
IT'S THE CELLS AND THE GLUE THAT
HOLDS THE CELLS TOGETHER.
THE EXTRACELLULAR MATRIX PLAYS A
MAJOR ROLE IN THE FUNCTIONALITY
OF THE TISSUE, INCLUDING THE
STIFFNESS AS I SHOWED IN ONE OF
THE SLIDES.
SO THAT'S CRITICAL HONESTLY
BECAUSE FOR THE CELLS TO SE SECE
THEIR OWN ECM, IT TAKES TIME.
SO CELLS WILL SECRETE THEIR OWN
ECM, BUT THAT IS A TIME PROCESS
FOR THAT TO ACTUALLY OCCUR AND
TO BUILD THAT THREE DIMENSIONAL
ORGANIZATIONAL STRUCTURE.
SO THAT IS WHY VERY EARLY ON, WE
BASICALLY WENT TO THE STRATEGY,
WE JUST INK WRAIT INCORPOR.
YOU'RE NOT GOING TO HAVE THESE
ORGANOIDS AROUND TOO LONG TO
ACTUALLY THEM SECRETE THAT ECM.
THAT'S CRITICAL, WHEN WE LOOK AT
THE INFECTIVITY FOR EXAMPLE OF
THOSE LUNG MODELS THAT I SHOWED,
WE USE LUNG-SPECIFIC ECM, AND WE
FIND THAT THAT'S CRITICAL FOR
THE MIGRATION OF THE NEUTROPHILS
THROUGH THE VIRAL RESPONSE.
AND THIS IS A PROGRAM THAT DAVE
HONE FROM DTRA HAS NOW PUT
TOGETHER, THIS PATNOS PROGRAM
FOR US TO LOOK AT VIRUSES AND
HOW THEY ACTUALLY PENETRATE THE
CELL MEMBRANES AND HOW THEY
ACTUALLY GET THROUGH ALL THOSE
CELL LAYERS, AND YOU KNOW, IT'S
OKAY TO GET THROUGH AN
EPITHELIAL CELL LAYER BUT WHEN
YOU DON'T HAVE ECM THERE, HOW
REALISTIC TO THE NATURAL
RESPONSE.
>> THAT'S WHY ALL MICRO TISSUES
ARE THREE WEEKS OLD.
THEY HAVE TO PRODUCE THEIR OWN
CARDIOMYOCYTE MATRIX.
BUT THAT'S ALSO CONTRIBUTED BY
THE STROMAL CELLS.
>> YEAH.
SO I THINK IT'S SO IMPORTANT TO
HAVE THAT ECM.
IT REALLY IS.
>> ABSOLUTELY.
>> SO THERE'S A QUESTION IN THE
CHAT THAT SAYS, HOW DO YOU MAKE
SURE THAT THE TISSUE MODELS
REPRESENT HUMAN PHYSIOLOGY?
SO IT'S TIED TO YOUR DISCUSSION
IN THE SENSE THAT, LIKE, WELL,
IF I WANT TO MAKE A LUNG TISSUE,
I NEED TO FIND SOME LUNG ECM
SOMEWHERE, WHETHER IT'S
COMMERCIALLY OR FIND A SOURCE,
AND THEN HOW -- WHAT SORT OF
VALIDATIONS DO WE NEED TO DO,
RIGHT?
I MEAN --
>> THAT'S A REALLY GOOD
QUESTION, BECAUSE THE QUESTION
IS REALLY, WHAT CAN YOU ACTUALLY
MEASURE IN THE HUMAN BODY THAT
YOU COULD TRANSLATE INTO YOUR
MODEL?
YOU CAN'T MEASURE -- IN THE
HUMAN BODY DB EPITHELIAL
RESISTANCE.
YOU CAN MEASURE THAT IN ISOLATED
EP
EPITHELIUM.
SO IN A NUMBER OF PARAMETERS,
YOU CAN DO A HEAD TO HEAD
COMPARISON NUMERICALLY,
QUANTITATIVELY IT'S VERY
LIMITED.
SO FOR A HEART, YOU CAN DO THE
ACTION PRO TENSION.
POTENTIAL.
CAN YOU DO IT PRIMARY.
IN THE HEART ITSELF AND IN THE
MODEL.
BUT WHAT ELSE?
SECRETED FACTORS, SO MARKER, BUT
YOU HAVE TO DOWNSCALE THAT OR
UPSCALE TO MATCH THE AMOUNT OF
BLOOD AND AMOUNT OF TISSUE IN
THE HUMAN BHO DE.
BODY.
SO THESE ARE VERY DIFFICULT
CALCULATIONS.
PUTTING SENSORS IN THE BODY TO
DETECT THINGS SO THAT IT REALLY
IS A LOT OF WORK TO BE DONE
THERE.
SO AT THE MOMENT, WE MOSTLY GO
FOR SINGLE CELL RNA SEQ OR SOME
SORT OF GENOMICS OROMIX ASSAY TO
FIGURE THAT OUT.
SO LOOK AT THE METABOLIC PROFILE
OR THINGS LIKE THAT.
BUT IT'S QUITE LIMITED, THE
NUMBER OF HEAD TO HEAD
COMPARISONS YOU CAN DO.
>> AND THEN ACCESS TO THIS HUMAN
DATA, RIGHT?
WHERE IS THIS DATA RESIDING AND
HOW DO YOU GET IT?
WHETHER IT'S PUBLIC DATABASES OR
WHETHER YOU HAVE COLLABORATORS
THAT HAVE GENERATED IT.
JEFF, YOU HAVE A BUNCH OF
QUESTIONS, IF YOU GO TO THE Q &
A BOX THERE'S A BUNCH OF
QUESTIONS SPECIFIC TO YOUR
MODEL.
IF YOU COULD ADDRESS THOSE, TYPE
IN THE ANSWER, THAT WOULD BE
GREAT.
>> THANK YOU, MARK.
CERTAINLY.
I SEE ONE QUESTION FROM DAVID
ABOUT COMBINATION, THAT'S A
GREAT QUESTION LOOKING AT THE
COMBINATION.
IT'S NOT SOMETHING WE'VE TESTED
YET BUT SOMETHING WE COULD
CERTAINLY DO IN OUR SYSTEM.
WE COULD DO THE TWO OF THEM IN
ISOLATION.
WE DIDN'T INCLUDE THE RITONAVIR
BECAUSE THAT'S REALLY A
PHARMACOKINETIC COMPOUND THAT'S
ADDED TO CONTROL LIVER
METABOLISM, BUT I THINK WE COULD
CERTAINLY LOOK AT COMBINATION
THERAPIES IN THE SYSTEM GIVEN
THE THROUGH PUT AND THE SORT OF
STATISTICAL NATURE THAT WE HAVE
HERE.
THAT'S A GREAT QUESTION.
I THINK ANOTHER QUESTION WAS
JUST REGARDING THE MOLNUPIRAVIR,
WHETHER WE USED THE PARENT
MOLECULE OR THE PRO DRUG.
WE USED THE PARENT MOLECULE.
SO I THINK THERE'S ALWAYS
THESE -- THESE SORT OF GET AT
THE QUESTION OF THE INTRODUCTION
OF THE COMPOUND AND HOW WE MIMIC
IN VIVO WITH THAT, AND THE FACT
THAT WE HAVE TWO ORAL DRUGS AND
ONE INJECTABLE DRUG AND HOW DO
WE SORT OF MIMIC THAT.
SO THINK, TONY ATALA MADE A
GREAT COMMENT EARLIER ON IN HIS
TALK THAT WE'RE STILL -- THERE'S
STILL A LOT OF WORK TO BE DONE
WITH THESE SYSTEMS.
WE'RE MAKING A LOT OF PROGRESS
BUT ULTIMATELY RECAPITULATING
THE IN VIVO MICROENVIRONMENT AND
PUTTING THESE TOGETHER IS
CRITICAL, HAVING THESE VARIOUS
COMPONENTS, SO TOWARD THAT END,
WE'RE ADDING THE IMMUNE
COMPONENT INTO OUR LUNG MODEL IN
COLLABORATION WITH BARDA, AND
WE'RE ALSO ADDING SENSORS SO WE
CAN MONITOR THE MICROENVIRONMENT
AND COMPARE TO IN VIVO.
AS CHRISTINE WAS TALKING ABOUT
EARLIER, WITH SOME OF THESE
SECRETED FACTORS LIKE THE
REACTIVE OXYGEN SPECIES.
>> THANK YOU.
I HAVE A QUESTION FOR JASON, AND
IT HAS DO WITH GOING BACK TO
PRIMARY CELLS, IPS CELLS.
SO YOU SHOW US WHERE YOU TRY TO
IMMORTGAGIZE
IMMORTALIZE YOUR CELLS TO GET
BETTER REPRODUCIBILITY, AND THE
QUESTION IS REALLY, SHOULDN'T WE
ACTUALLY LEARN HOW TO HANDLE
VARIABILITY, ESPECIALLY FROM
PATIENT TO PATIENT INSTEAD OF
TRYING TO ENGINEER CELLS SO WE
GET THE TECHNICAL
REPRODUCIBILITY?
HOW DO YOU GUYS HANDLE THESE
KIND OF DECISIONS?
>> SO MY UNDERSTANDING IS KIND
OF LIKE -- THEY ARE JUST TOOLS.
FOR EXAMPLE, SOME PATIENT
DRUG -- AND THEN YOU FIND THE
SPECIFIC GENE THAT MUTATED.
THERE IS SPECIFIC MUTATION IN
ENDOTHELIAL CELLS.
YOU CAN ACTUALLY INDUCE THOSE
MUTATIONS IN THE -- CELLS AND
THEN YOU CAN USE THOSE AS A TOOL
TO STUDY FUNCTIONALLY HOW THOSE
MUTATIONS ACTUALLY CAUSE THOSE
DYSFUNCTION.
SO IT IS A TOOL BUT IT'S NOT
ALL -- IT'S NOT -- SO WE ALWAYS
NEED TO FIND KIND OF DIFFERENCES
FROM THE PATIENT AND THEN JUST
TO VALIDATE USING THE CELL
LINE -- THAT'S MY UNDERSTANDING.
>> SO I GUESS IT GOES BACK TO
THE POINT CHRISTINE WAS MAKING,
IT REALLY DEPENDS ON WHAT YOU
WOONT TO
WANT TO USE THE SYSTEM FOR AND
WHAT CELLS YOU USE, BECAUSE
JEFF, YOU SHOWED BEAUTIFUL --
CVs IN YOUR SYSTEM AND THAT
WAS FROM PATIENT TO PATIENT, YOU
REALLY DIDN'T SEE MUCH VARIATION
IN TERMS OF INFECTIVITY.
SO -- AND THOSE WERE PRIMARY
CELLS, SO IT'S PRETTY AMAZING AT
LEAST FOR THOSE PRIMARY CELLS,
EPITHELIAL CELLS HOW ROBUST THE
SYSTEM IS, AND REALLY
TECHNICALLY, PHYSIOLOGICALLY,
YOU SEEM TO BE READY FOR PRIME
TIME SCREENING.
>> IT IS A VERY ROBUST SYSTEM.
I THINK SOME OF WHAT WOULD
REALLY BE INTERESTING AND WOULD
PROBABLY INTRODUCE MORE
VARIABILITY IS IF WE START TO
LOOK AT PARTICULAR COMORBIDITIES
THAT WE KNOW DRIVE DIFFERENT
DIFFERENTIAL RESPONSES.
SO IF WE START LOOKING AT
ASTHMATICS OR OTHER
COMORBIDITIES THAT MIGHT DRIVE
DIFFERENCES IN RESPONSE, THAT'S
WHAT WE MIGHT SEE.
I THINK WITH THE RESEARCH BRON
BRONCHOSCOPIES, A LOT OF THE
DONOR POPULATIONS ARE VERY
SIMILAR.
IT'S A SIMILAR PATIENT
POPULATION, USUALLY YOUNGER,
HEALTHIER PEOPLE THAT RESPOND TO
THE HOSPITAL SURVEYS OR REQUESTS
TO DONATE TISSUE WITH A
BRONCHOSCOPY, BUT I THINK
BROADENING THAT TO PATIENTS WITH
VARIOUS DISEASE PROFILES WOULD
REALLY BE INTERESTING.
>> THERE WAS A QUESTION IN THE
CHAT, I THINK CHRISTINE
MENTIONED THAT SHE WANTED TO
ANSWER IT.
IT WAS AN INTERESTING QUESTION
GOES BACK TO THE ISSUE OF
VARIABILITY, IN THIS CASE ARE
THE ECM DONOR TO DONOR.
SO VARIABILITY IN THE ECM FROM
DONOR TO DONOR.
WE'RE NOT TALKING ABOUT CELLS
HERE, WE'RE TALKING ABOUT ECM.
AND I GUESS I DIDN'T SEE LINDA,
I DON'T THINK SHE WAS ABLE TO
MAKE IT NOW, BUT HOW THAT KIND
OF --
>> I PRESSED THE WRONG BUTTON
ACTUALLY.
I'M QUITE HAPPY TO GIVE A GO.
BUT IT'S REALLY FOR -- NOT FOR
ME, IT'S MORE FOR ANTHONY.
THERE ISN'T THAT MUCH
VARIABILITY FROM DONOR TO DONOR
IN THE EXTRACELLULAR MATRIX, AND
IT'S NOT REALLY LOGICAL IT WOULD
BE HUGE, BECAUSE A NORMAL ORGAN
HAS TO FUNCTION.
OFTEN IF AN ORGAN IS FIBROTIC,
IT WILL VARY FROM DONOR TO DONOR
AND IT'S, OF COURSE, QUITE HARD
TO SEE HOW GUY FIBROTIC IT IS.
IT WOULD DEPEND ON THE AGE OF
THE DONOR AND THINGS LIKE THAT.
>> TONY?
>> THANK YOU, MARC, AND THANK
YOU, CHRISTINE.
SO BASICALLY, THE IMPORTANT
THING, OFTEN, IS ORGAN-SPECIFIC
ECM, RIGHT?
BECAUSE ECM FROM THE LIVER IS
NOT GOING TO BE THE SAME AS THE
EYM FROM THE LUNG.
CM FROM THE LUNG.
ONCE YOU GET DOWN TO THAT LEVEL,
THE WAY WE DO IT IS WE TAKE THE
ORGAN, WE USE -- TO WASH -- AT
WAY, WE'RE LEFT JUST WITH THE
EXTRACELLULAR MATRIX, WE MAKE
THE MATRIX INTO A POWDER AND WE
THEN RECONSTITUTE THAT POWDER TO
CREATE THE STRUCTURE AND THEN WE
MATCH THE STIFFNESS TO THE
NORMAL STIFFNESS THE ORGAN WOULD
HAVE.
AGAIN JUST AS A WAY TO MAKE SURE
WE REPLICATE AS MANY OF THE
PROPERTY AS POSSIBLE IN THAT
ORGANOID SYSTEM AS YOU WOULD SEE
IN A NORMAL HUMAN TISSUE AND
ORGAN.
BECAUSE EVERYTHING PLAYS A ROLE.
AND THERE IS NO VAST VARIABILITY
BETWEEN PATIENTS BECAUSE AS YOU
KNOW, IT DOESN'T HAVE ANY IMMUNE
PROPERTIES, IT'S JUST THE ECM,
WHICH IS MAINLY COLLAGEN, MOSTLY
TYPE I COLLAGEN, ALSO TYPE 2,
TYPE 3, SOME LAMININ.
BUT IT VARIES FROM ORGAN TO
ORGAN.
>> I THINK THE POINT IS THE
BATCH TO BATCH VARIABILITY.
SO IF YOU TAKE ONE LIVER, IS
THIS -- THE BATCH THE SAME AS
THE OTHER LIVER.
WE KNOW WITH MATRIGEL, IT IS
BATCH TO BATCH VARIABILITY EVEN
THOUGH THEY'RE MADE IN THE SAME
WAY.
SO I THINK THIS IS THE QUESTION,
OF COURSE YOU GET HUGE BATCHES
FROM A HUMAN LIVER, BUT DO YOU
SEE BATCH TO BATCH DIFFERENCES?
>> YOU DO GET THIS HUGE AMOUNT,
OF COURSE, OF THE ECM THAT
ALLOWS YOU TO REALLY -- TEST
REALLY MILLIONS OF THESE
ORGANOIDS.
WITH THE SAME MATRIX.
>> I'M GOING TO GIVE AN
OPPORTUNITY TO THE PANELISTS
THAT ARE NOT THE SPEAKERS TO ASK
ANY QUESTIONS, IF THEY HAVE ANY?
SARINE, SIMON, JONI, ANY
PRESSING QUESTIONS?
>> I GUESS I'D JUST LIKE TO
QUICKLY GO BACK TO THE QUESTION
ABOUT VARIABILITY OF DIMERS.
IN TERMS OF SCALE, ECM, YOU
MATCH THE BATCH AND BE ABLE TO
USE THAT, BUT I THINK AT THE
MOMENT, EVERYBODY IS SAYING WHAT
THEY WANT TO USE FRESH TISSUE,
FRESH PRIMARY TISSUE FROM
PATIENTS, BUT HOW DO WE MOVE
AWAY FROM THAT INTO A SYSTEM
WHERE ANYBODY CAN ACCESS A
CERTAIN LOT AND DO THE SAME
EXPERIMENT, NO MATTER WHERE THEY
ARE ON THE PLANET, AND THEN
WHERE THEY ARE IN TIME FROM THE
DATE THAT THOSE CELLS WERE
TAKEN.
AND I DON'T THINK WE'VE GOT
THERE YET.
I'VE TALKED ABOUT PLURIPOTENTIAL
STEM CELLS AND ALL I'VE HEARD IS
THAT'S PROBLEMATIC, BUT HOW DO
WE MOVE FORWARD FROM THIS
SITUATION?
>> SIMON, THAT'S A GREAT
QUESTION.
THAT'S THE BILLION DOLLAR
QUESTION.
BECAUSE THAT'S WHAT WE WERE
FACING.
THAT LAST SLIDE THAT I SHOWED
WHICH WAS BASICALLY THE
VARIABILITY.
THAT'S THE CHALLENGE.
I ALWAYS LIKE TO SAY, YOU KNOW,
HOW IS IT POSSIBLE THAT WE COULD
DETECT THAT TOXICITY FROM THOSE
DRUGS SO READILY IN OUR SYSTEM
WHEN IT WAS NOT POSSIBLE IN THE
2D MODELS, IT WAS NOT POSSIBLE
IN THE ANIMALS, IT WAS NOT
POSSIBLE IN THE HUMAN CLINICAL
TRIALS AND THEN 11 YEARS LATER,
YOU GET THIS -- YOU FINALLY FIND
OUT THIS DRUG IS CAUSING
PROBLEMS, OR THREE YEARS LATER,
AND SO I THINK THERE'S A LITTLE
BUILT OF A -- LET ME DISCUSS
THIS BALL ABC A LITTLE Y
THEORY ON THIS IS THAT, YOU
KNOW, WHEN WE ARE LOOKING AT THE
ACTUAL DRUG TO THE ORGAN, WE ARE
REALLY GETTING RID OF ALL THE
NOISE.
WE'RE GETTING RID -- I SHOULD
SAY I SHOULD BE MORE SPECIFIC,
WE'RE GETTING RID OF A LOT OF
THE NOISE, BECAUSE WHAT HAPPENS
IN THE HUMAN IS WE ALL
METABOLIZE DRUGS DIFFERENTLY,
WE'RE ALL GENETICALLY DIFFERENT,
ALL EPIGENETICALLY DIFFERENT, WE
HAVE DIFFERENT DIETS, WE PROCESS
OUR FOODS DIFFERENT, WE HAVE
DIFFERENT METABOLISM RATES, AND
ALL THESE FACTORS GO INTO THAT
RESPONSE.
THAT IS WHY IT TOOK 11 YEARS FOR
THERE TO BE ENOUGH CASES WHERE
PEOPLE COULD CONNECT THE DOTS
AND SAY THIS DRUG IS CAUSING
HEART BLOCK, AND YET YOU PUT
INTO THE SYSTEM AND THERE IT IS,
RIGHT?
IT'S LIKE NINE DAY, THERE IT IS.
AND I THINK SO THERE ARE
ADVANTAGES TO GETTING RID OF ALL
THE NOISE BECAUSE YOU'RE REALLY
LOOKING AT THE EFFECT OF THE
DRUG AGAINST THE TISSUE AND THE
ORGAN AND YOU'RE GETTING RID OF
A LOT OF THE NOISE.
ON THE OTHER HAND, THERE ARE
MANY SITUATIONS WHERE YOU WANT
TO PUT THAT NOISE BACK IN,
RIGHT?
YOU WANT TO PUT THAT NOISE BACK
IN, LIKE THE PERSONALIZED
MEDICINE APPROACH LIKE THE
TUMORS THAT WE'RE TALKING ABOUT.
BECAUSE EACH TUMOR IS DIFFERENT,
IT'S GOING TO HAVE DIFFERENT
EPIGENETIC CHANGES AND WE'RE
GOING TO TAKE THAT TUMOR FROM
THAT PATIENT, WE'RE GOING TO
TEST THAT TUMOR AGAINST SPECIFIC
CHEMOTHERAPY, YOU'RE IDENTIFYING
THAT TUMOR WITH THEIR OWN
GENETIC AND EPIGENETIC CHANGES
AND YOU REALLY WANT THAT DATA IN
TERMS OF WHAT IT DOES.
SO I THINK WE HAVE TO MINIMIZE
BATCH TO BATCH VARIABILITY, WE
HAVE TO WORK ON BETTER AND MORE
RELIABLE CELL SOURCES, WE HAVE
TO WORK ON STANDARDIZATION OF
THE SYSTEMS, BUT ALSO TO KEEP IN
MIND WHAT THE ACTUAL END POINT
IS OF WHAT YOU'RE LOOKING FOR.
>> IF I COULD JUST JUMP IN
THERE, THE INTERNATIONAL -- STEM
CELL RESEARCH IS WORKING EXACTLY
ON THAT ISSUE, SO WORKING ON
STANDARDIZATION DOCUMENTS, HOW
TO STANDARDIZE STEM CELLS.
AND IT WAS ADULT STEM CELLS AND
IPS CELLS AND EMBRYONIC STEM
CELLS.
WHAT YOU NEED TO MONITOR IN
TERMS OF GENETIC DRIFT,
PROPERTIES, ALL THE KINDS OF
THINGS THAT PEOPLE HAVE TROUBLE
WITH, WHAT DO YOU HAVE TO DEFINE
TO ALLOW THE FIELD TO BE ABLE TO
DO THINGS REPRODUCIBLY FROM LAB
TO LAB, PERSON TO PERSON.
I THINK WE'RE GETTING THERE,
PARTICULARLY WITH COMMERCIAL
COMPANIES PRODUCING REAGENTS IN
LARGE SCALE GOING FROM GROWTH
FACTORS TO -- MOLECULAR BASED
CULTURE METHODS, THINGS LIKE
THAT.
SO IT IS GETTING THERE.
BUT WE WILL HAVE A CONSENSUS
STATEMENT DOCUMENT BEFORE THE
END OF THE YEAR DEFINING WHAT
YOU NEED TO SAY ABOUT THOSE STEM
CELLS.
AND I AGREE THAT OF COURSE THERE
ARE A LOT OF DIET ISSUES AND
LIFESTYLE ISSUES THAT IMPACT
CELLS, BUT AT LEAST I THINK WITH
SOME OF THE STEM CELL MODELS, WE
CAN ADDRESS THE GENETIC ISSUE,
PERHAPS EVEN THE EPIGENETIC
ISSUES AND AT LEAST GET THAT
STANDARD.
AND PARTICULARLY ADDRESS
VARIANTS OF UNKNOWN
SIGNIFICANCE, SMALL MACULAR
CHANGES IN THE GENOME THAT ARE
CAUSING PERSON TO PERSON
VARIABILITY.
BEST CASE SCENARIO, DRUGS WORK
NOW IN 1 IN 4 PATIENTS, THE TOP
10 PRESCRIBED DRUGS IN THE U.S.
AND THE WORST CASE, IN THOSE 10
TOP PRESCRIBED DRUGS, THERE ARE
1 IN 25 PATIENTS RESPONDING AS
EXPECTED.
THAT'S TERRIBLE.
THAT'S AN ENORMOUS WASTE OF
RESOURCES.
SO THERE'S A LOT OF GAIN TO BE
HAD, EVEN IDENTIFYING NOT JUST
FOR TUMORS BUT WHO'S GOING TO
RESPOND TO EVEN A RELATIVELY
EXPENSIVE DRUG, IF IT'S A DRUG
USED BY MILLIONS OF PEOPLE.
>> TECHNICALLY WE SHOULD END
NOW, BUT IF THE SPEAKERS ARE
WILLING TO STAY A LITTLE LONGER,
THERE'S JUST A COUPLE OF
QUESTIONS THAT HAVE COME UP AND
SOME OF THEM ARE RELATED TO THIS
DISCUSSION IN TERMS OF
STANDARDIZATION OF MPSs AND
WHAT DO WE MEAN BY
STANDARDIZATION.
WE'VE TALKED ABOUT VALIDATING
THE TISSUES ABOUT ALL THE OMICS
TO MAKE SURE THEY LOOK LIKE
NATIVE, THAT'S A HALLMARK IN
ITSELF.
BUT THEN WE'RE TALKING PLIEK LIY
WAS SAYING ABOUT REMOVING THE
NOISE MAYBE TISSUE BY TISSUE,
LOOKING AT TOXIC EFFECTS,
LOOKING AT EFFICACY, MAYBE
HAVING ANOTHER LEVEL WHERE YOU
ACTUALLY LOOK AT MAYBE
METABOLISM IN A PARTICULAR DRUG.
SO THE QUESTION IS, IS THERE
GOING TO BE A STANDARDIZED MODEL
WHICH POTENTIALLY DECREASES
VARIATION AMONG DIFFERENT
MODELS? I DON'T KNOW WHO WANTS
TO TAKE THESE QUESTIONS.
THE WAY I SEE THE FIELD GOING IS
WE HAVE MANY DIFFERENT
TECHNOLOGIES.
A LOT OF IT BECAUSE HAVE BEEN
MENTIONED IS REALLY DEPENDING --
IT'S A FIT-FOR-PURPOSE,
DEPENDING ON WHAT APPLICATION
YOU'RE LOOKING FOR, SO MAYBE I'M
WRONG, I DON'T SEE ONE
STANDARDIZED MPS MODEL, BUT
TONY, YOU UNMUTED YOURSELF.
YOU WANT TO TAKE THIS ON?
>> ACTUALLY, I JUST UNMUTED JUST
IN CASE I WAS READY, BUT I'LL BE
HAPPY TO GO FIRST.
I THINK WHAT WE REALLY NEED DO
IS START LOOKING AT THE FUTURE
OF THE FIELD IN TERMS OF CELL
SOURCING AND CELL
STANDARDIZATION AND THE GENETIC
COMPONENTS OF THE CELLS.
THAT'S REALLY WHAT WE NEED TO
DO.
WE NEED TO START BUILDING BANKS.
AS YOU KNOW, IT ABSOLUTELY NO
DIFFERENT WHEN WE'RE TALKING
ABOUT A STEM CELL BANK.
BASICALLY IT'S NOT LIKE WE'RE
RE-EXPLORING THE FIELD BECAUSE
WE KNOW THAT IF WE HAVE A UNIQUE
SET OF ABOUT 7,000 STEM CELL
POPULATIONS, THAT WOULD PROVIDE
OVER 90% OF CELL POPULATION WITH
A PERFECT GENETIC MATCH, SO
THERE ARE WAYS TO DO THIS, BUT
THEY HAVE TO BE UNIQUE.
ALL THIS HAS BEEN WORKED OUT FOR
THE CELL BANKING INDUSTRY,
RIGHT, FOR THE STEM CELL
BANKING.
YOU LOOK AT BONE MARROW BANKING
AND YOU LOOK AT CORE BLOOD
BANKING, WHAT IS THAT MAGIC
NUMBER THAT GIVES YOU THAT
GENETIC VARIABILITY.
SO THE MAIN THING REALLY IS TO
ACTUALLY START ESTABLISHING SUCH
A BANK THAT WILL GIVE US THIS
GENETIC VARIABLE SO WE CAN THEN
START TO PICK OUT THE THINGS
THAT WE WANT TO LOOK AT FROM
THESE BANKS.
THEN WE HAVE THIS RESOURCE
THAT'S AVAILABLE TO EVERYBODY,
AND IT WOULD BE NICE DO IT WITH
CELLS WHICH ARE ALREADY, YOU
KNOW, MORE IN A DIFFERENTIATED
PATHWAY REALLY.
THE CHALLENGE AS YOU KNOW IS
GETTING TO THAT DERMAL
DIFFERENTIATION.
MANY TIMES YOU CAN ONLY DO THAT
WITH A PRIMARY HUMAN CELL.
I KNOW IT'S HARDER TO DO, BUT IF
YOU DON'T START NOW, THEN WE'RE
GOING TO BE LOOKING AT THE SAME
PROBLEM 20 YEARS FROM NOW.
SO WE MIGHT AS WELL BITE THE
BULLET, REALIZE WHAT WE NEED TO
DO, AND JUST GO AFTER IT.
>> AWESOME.
ANYBODY ELSE HAVE ANY COMMENTS?
>> THERE'S ALSO ENGINEERING
SOURCES OF VARIABILITY THAT I
THINK WE NEED TO ADDRESS, AND
ONE OF THE CHALLENGES, I THINK,
IN THE ORGAN ON A CHIP FIELD HAS
BEEN THAT IT'S SOMETIMES VIEWED
AS SYNONYMOUS WITH THESE KIND OF
USB STICK KIND OF IMAGE, WHICH
IS REALLY KIND OF A SMALL SCALE,
IT'S VERY POWERFUL IN TERMS OF
THE FIDELITY OF SOME OF THESE
MODELS.
YOU CAN DO A LOT OF COMPLEX SORT
OF ENGINEERING IN THESE SYSTEMS,
BUT IT DOES REPRESENT A
LIMITATION RELATIVE TO THE GOLD
STANDARD HIGH THROUGHPUT MICRO
TITER PLATES THAT THE INDUSTRY
IS USED TO WORKING WITH.
WE WANT TO ELIMINATE THOSE
SOURCES OF ENGINEERING
VARIABILITY BATCH TO BATCH,
PLATE TO PLATE, PROVIDE THE
THROUGHPUT PROFESSOR GRIFFITH
TALKED EARLIER ABOUT THE ISSUES
WITH DRUG GET AWAY FROM
MATERIALS THAT CAUSE THESE
PROBLEMS, IMPLENTY SENSORS SO WE
KNOW WHAT'S GOING ON IN THESE
SYSTEMS.
THAT'S WITH THE ENGINEERING CAN
COME IN AND HELP ADDRESS SOME OF
THOSE PROBLEMS SO WE AREN'T
DEALING WITH THOSE ISSUES AS
WELL AS THE NATURAL BIOLOGIC
VARIATION.
>> THAT'S A GOOD POINT.
I THINK THE OTHER ASPECT OF HOW
YOU GET THESE TECHNOLOGIES
ARE -- SO THAT BUYOLOGIST CAN
USE IT AS WELL.
I THINK CHRISTINE HAS TO LEAVE.
THANK YOU SO MUCH, CHRISTINE.
THANKS SO MUCH FOR
PARTICIPATING.
IT'S BEEN GREAT.
SO THE QUESTION OF COMONETIZING
THE TECHNOLOGY AND GETTING TO
NOT ONLY THE BIOENGINEERING LABS
BUT BIOLOGY LABS SO THEY CAN USE
IT AT A REASONABLE COST.
SO THESE TECHNOLOGIES ARE VERY
EXPENSIVE.
SOMETIMES THAT'S A LIMITATION,
IF A BIOLOGY LAB WANTS TO USE
ONE OF THESE -- CHIPS AND THEY
CAN ONLY AFFORD LIKE ONE OR TWO
CHIPS, THERE'S SO MUCH BIOLOGY
YOU CAN DO.
I THINK THEY'RE GOING TO KICK US
OUT OF THE ROOM.
I JUST GOT A NOTE TBR FROM AB.
SO SARINE, I THINK WE'RE GOING
TO CLOSE HERE.
I KNOW THERE'S A FEW MORE
QUESTIONS IN THE CHAT, BUT --
>> YEAH, NO WORRIES.
>> I THINK WE ARE --
>> THANK YOU.
THANK YOU SO MUCH, MARC, AND
THANK YOU, ALL THE SPEAKERS.
THIS WAS AN AMAZING SESSION, AND
THE DISCUSSION WAS JUST AMAZING.
I JUST COULDN'T -- I DON'T WANT
TO STOP IT, BUT WE ALL HAVE TO
GO.
SO AGAIN, MANY THANKS.
I KNOW THERE'S MANY QUESTIONS
LEFT.
WE TRIED TO ANSWER AS MANY
QUESTIONS AS POSSIBLE.
IF YOU SEE A QUESTION THAT YOU
CAN ANSWER BRIEFLY BY TYPING, GO
AHEAD AND DO SO.
APOLOGIES FOR THE PEOPLE THAT WE
COULDN'T ANSWER ALL THE
QUESTIONS, BUT I'M SO GLAD THAT
YOU ASKED THE QUESTIONS.
I HOPE YOU ENJOYED THE
DISCUSSION.
SO THANK YOU AGAIN.
BEFORE I WRAP TODAY'S SESSION, I
WOULD LIKE TO SAY THAT WE HAVE
TWO AWESOME SESSIONS TOMORROW,
AND WE WILL RECONVENE TOMORROW
AT 11:00 A.M., SO HOPEFULLY YOU
ENJOYED TODAY, YOU LEARNED A
LOT, AND I WILL SHARE HERE THE
AGENDA FOR TOMORROW QUICKLY.
FOR FOLKS TO KNOW WHAT'S COMING
TOMORROW.
OKAY.
SO TOMORROW WE WILL START AT
11:00, AND WE WILL HAVE THE
SECOND SESSION WHICH IS GOING TO
BE ON UTILITY OF THE EXISTING 3D
TISSUE MODELS FOR ANTIVIRAL DRUG
DEVELOPMENT.
AND THEN WE WILL HAVE A THIRD
SESSION, SESSION 3, IT WILL BE
ON USE OF ROBUST AND
REPRODUCIBLE 3D TISSUE MODELS
FROM DRUG DISCOVERY THROUGH
DEVELOPMENT, AND WE WILL WRAP UP
TOMORROW WITH A FINAL SESSION,
SESSION 4, CLOSING SESSION THAT
WE'LL DISCUSS SUMMARY OF
DISCUSSIONS AND PERSPECTIVES ON
THE CHALLENGES AHEAD.
SO AGAIN, HOPEFULLY YOU ENJOYED
TODAY.
THANK YOU FOR ALL THE SPEAKERS.
THIS WAS AMAZING.
WE WILL HOPEFULLY SEE YOU ALL
TOMORROW AT 11:00.
MANY THANKS AGAIN.
HAVE A WONDERFUL --
>> THANK YOU, TONY, JASON, LINDA
AND CHRISTINE, GREAT TALKS AND
GREAT DISCUSSION.
APPRECIATE IT.