GOOD MORNING.
I ALSO WANT TO SAY GOOD
AFTERNOON AND GOOD EVENING TO
SOME OF YOU WHO LIVE IN THE
DIFFERENT PART OF THE GLOBE.
SO GOOD TO SEE YOU ALL HERE
AGAIN.
WELCOME TO DAY 2 OF THE WORKSHOP
ON PROGENITOR RESILIENCE AND THE
EARLY SO ONSET OF CHRONIC LUNG
DISEASE.
ON BEHALF OF THE ENTIRE WORKSHOP
PLANNING TEAM, I WOULD LIKE TO
WELCOME YOU AND THANK YOU FOR
YOUR PARTICIPATION.
I'M DR. QING LU, PROGRAM
OFFICIAL IN THE DIVISION OF LUNG
DISEASE.
WHAT A GREAT MEETING WE HAD
YESTERDAY, RIGHT?
MANY PEOPLE HAVE SAID THAT.
I'M VERY EXCITED AND HAPPY TO
REPORT THAT IN THE AVERAGE
YESTERDAY, WE HAD AROUND 200
PEOPLE JOIN THE ZOOM ROOM AND
ABOUT 20 PEOPLE JOINED THE NIH
VIDEOCAST PLATFORM AT A GIVEN
TIME.
SO THAT SPEAKS THE INTEREST OF
THE COMMUNITY TO THIS WORKSHOP.
I WOULD LIKE TO THANK OUR
CO-CHAIRS, ALL THE MODERATORS
AND SPEAKERS, AND TH THE NIH
WORKSHOP SUPPORT TEAM.
BECAUSE OF YOU, THIS HAPPENED.
WE WOULD LIKE TO LOOK FORWARD TO
THE SUCCESSFUL DAY TWO OF THIS
WORKSHOP, AND ALSO WELCOME YOU
TO ACTIONABLE RECOMMENDATIONS.
NOW LET ME WELCOME MYCO-LEAD,
DR. CHRISTIAN GOMEZ, TO SAY A
FEW WORDS BUT ALSO A RECAP FOR
DAY ONE.
THANK YOU.
CHRISTIAN, TAKE IT OVER.
>> THANK YOU.
THANK YOU SO MUCH, QING LU.
WELCOME AGAIN TO THIS EXCITING
WORKSHOP.
AS SHE SAID, WE HAVE A TERRIFIC
DAY YESTERDAY, AND MANY THINGS
HAPPENED, YOU KNOW, THAT NEED TO
BE PUT BACK FOR THOSE THAT WERE
PRESENT THERE AND FOAS JOINING
US TODAY FOR THE FIRST TIME.
SO WITHOUT FURTHER PREAMBLE, I
WOULD LIKE TO INTRODUCE
DR. SEUNGHYE HAN FROM
NORTHWESTERNORTHWESTERN UNIVERS.
SHE IS GOING TO PRESENT THE
RECAP FOR DAY ONE.
DR. HARNTION PLEAS HAN, PLEASE D
ENJOY THIS EXCITING DAY.
THANK YOU.
>> HI, CHRISTIAN.
THANK YOU FOR THE KIND
INTRODUCTION.
SO YESTERDAY WE HAD A VERY
EXCITING WORKSHOP.
OUR WORKSHOP WAS OPEN BY KEYNOTE
SPEAKER DR. RAVI KALHAN WHO
DISCUSSED HOW TO DEFINE
RESPIRATORY RESILIENCE AND
PROMOTE LUNG HEALTH.
HE EMPHASIZED THAT WE NEED A
FRAMEWORK FOR BIOMARKER
DEVELOPMENT SIMILAR TO
CHOLESTEROL-LIKE INDICATOR FORCE
LUNG HEALTH.
TO DO THIS, WE NEED TO SHIFT OUR
FOCUS FROM DISEASE DEVELOPMENT
TO UNDERSTANDING THE MECHANISMS
OF RESPIRATORY HEALTH AND
RESILIENCE.
IN SESSION 1, WE HAD FOUR TALKS
ON THE ROLE OF STEM PROGENITOR
CELLS IN CHRONIC LUNG DISEASE.
FIRST WE DISCUSSED THE
MITOCHONDRIA INTEGRATED STRESS
RESPONSE, ISR, THAT CONTROLS
STEM CELL FATE.
INDEPENDENT OF ATP SYNTHESIS,
MITOCHONDRIAL DYSFUNCTIONS,
SPECIFICALLY A HIGH RATIO,
PREVENTS TRANS SIGGAL CELLS FROM
DIFFERENTIATING INTO -- HOW THE
ISR CONTROL THE CELL FATE AND
WHETHER LEADS TO LUNG DISEASE
NEEDS TO BE FURTHER INVESTIGATED
WITH THE GOAL OF IDENTIFYING
TARGETS TO IMPROVE THE WASTE AND
PROGENITOR CELLS RESPONSE TO
STRESS TO ENHANCE THEIR
RESILIENCE.
NEXT WE TALKED ABOUT DIGITAL
AIRWAY PROGENITOR CELLS IN HUMAN
LUNG DISEASE.
THEY'VE BEEN IDENTIFIED IN HUMAN
AND FERRET LUNGS.
ALTERATION IN THIS CELL
POPULATION ARE ASSOCIATED WITH
COPD.
FURTHER STUDIES ARE NECESSARY TO
UNDERSTAND THE RESPIRATORY
AIRWAY PATHOPHYSIOLOGY AND
PROMOTE LUNG RESILIENCE.
THE THIRD TALK WAS ABOUT A NEW
AIRWAY CELL TYPE, HILLOCKS.
FUTURE STUDIES SHOULD FURTHER
CHARACTERIZE THE FUNCTION OF
HILLOCK CELL AND THEIR
CONTRIBUTIONS TO AIRWAY
RESILIENCE.
THE FINAL PRESENTATION IN
SESSION ONE WAS ON DIFFIC
DIFFERENTIATION AND IMPACT OF
PERTURBING TRANSITIONAL CELL
STATUS ON TISSUE REPAIR.
COMMON INJURY REPAIR PROCESSES
MAY UNDERLIE BOTH EMPHYSEMA AND
FIBROSIS, REPRESENTING TWO
EXTREMES OF THE SAME INJURY
RESPONSE.
FURTHER RESEARCH IS NEEDED FOR
ELUCIDATE THE MOLECULAR
MECHANISM AND THEIR ROLE IN
RESILIENCE FOR DISEASE
SUSCEPTIBILITY.
SO NEXT SESSION TWO FOCUSES ON
EMERGING TECHNOLOGIES AND MODEL
SYSTEMS TO STUDY LUNG
RESILIENCE.
FIRST WE DISCUSSED THE RECENT
ADVANCES IN ORGANOID SYSTEMS.
VASCULARIZED ORGANOIDS --
TRACING IS A VALUABLE TOOL TO
PRECISELY LABEL BRONCHIAL STEM
CELLS TO UNDERSTAND THE ORIGINS
OF A T2 CELLS IN HOMEOSTASIS AND
DISEASE STATE.
ON AIRWAY CULTURE SYSTEM
INCLUDES COMMON IN RARE AIRWAY
CELL TYPES THAT MIMIC THE ION
TRANSPORT DEFECTS OF CYSTIC
FIBROSIS.
PROVIDING A VALUABLE PLATFORM
FOR PATHOGEN STUDIES --
SCREENING AND UNDERSTANDING
BIOPATHOLOGY.
LASTLY WE DISCUSSED THE VALUE OF
FERRET MODELS TO STUDY LUNG
DISEASE.
DUE TO THE ANATOMICAL AND
CELLULAR DIFFERENCES BETWEEN
HUMAN AND RODENT AIRWAY
STRUCTURE, TRANSGENIC FERRETS
ARE AN IDEAL MODEL TO STUDY
HUMAN SPECIFIC LUNG DISEASE AND
RESPIRATORY HELP.
WE DISCUSSED THE LIMITATIONS OF
EXISTING MODELS, EMPHASIZING THE
NEED FOR LONGITUDAL HUMAN
STUDIES AND ADVANCED MODELS THAT
BETTER REFLECT HUMAN RESPIRATORY
HEALTH AND TRAJECTORIES TO
UNDERSTAND THE RESILIENCE
MECHANISMS.
IN SESSION THREE, WE DISCUSSED
THE PROGENITOR BASED MARKERS
THAT PREDICT EARLY ONSET OF
CHRONIC
DYNAMICS.
A SPECIFIC PROTEIN MARKER WAS
IDENTIFIED.
THE SESSION CONCLUDED WITH A
DISCUSSION ON A LARGER SCALE
DATA APPLICATIONS IN PREDICTIVE
MODELS AND THE VALUE OF
LONGITUDAL DATA TO DEVELOP
BIOMARMARKERS TO PREDICT LUNG
HEALTH AND RESILIENCE.
SO I TRIED TO CONCISE ALL THE
WONDERFUL POINTS TO THIS VERY
SHORT, CONCISE -- SO I MIGHT
MISS SOME THINGS BUT I EXPECTED
TO IS GOING TO BE MORE WONDERFUL
DISCUSSION AND BY AMAZING
SPEAKERS.
SO WE CAN -- I'LL GO TO THE
MODERATORS OF THE SESSION FOUR,
DR. CHAPMAN AND DR. MAJKA.
>> HI, GOOD MORNING.
I'M SUE MAJKA, AND IT'S MY
PLEASURE INTRODUCE DR. ANDREW
WILSON AS OUR FIRST SPEAKER THIS
MORNING.
HE IS FROM BOSTON UNIVERSITY,
THE CENTER FOR REGENERATIVE
MEDICINE, AND THE SCIENTIFIC
DIRECTOR OF THE ALPHA-1
FOUNDATION.
>> HELLO, THANK YOU VERY MUCH,
DR. MAJKA.
I'M LOOKING FORWARD TO THE DAY.
IT'S MY PLEASURE TO BE ABLE TO
SPEAK TO YOU TODAY ABOUT THE
APPLICATION OF PATIENT IPS CELLS
TO MODEL INA VERY VO LUNG
BIOLOGY AND SPECIFICALLY TO
MODEL THAT BIOLOGY AS IT RELATES
TO COPD AND EMPHYSEMA, THE
DISEASES MY LAB PRIMARILY
FOCUSES ON.
SO I WANT TO START OFF JUST BY
MAKING A COUPLE OF OBSERVATIONS
THAT I THINK INDIVIDUALLY ARE
NOT GOING TO BE A SURPRISE TO
THIS AUDIENCE, BUT THAT
COLLECTIVELY UNDERLIE THE
RATIONALE FOR THE APPROACH THAT
WE'VE TAKEN TO ADDRESS COPD AND
EMPHYSEMA IN OUR WORK.
THE FIRST OF THESE OBSERVATION
WAS IN PART MADE PRETTY WELL
YESTERDAY SO I WON'T BELABOR IT
BUT JUST THE OBSERVATION THAT
HUMAN LUNGS DIFFER STRUCTURALLY
FROM RODENT LUNGS IN WAYS THAT
CAN BE REALLY SALIENT AS RELATES
TO MODELING COPD AND EMPHYSEMA.
BUT I'D LIKE TO EXTEND THAT
POINT TO THE GENETICS OF THE
INDIVIDUAL CELL TYPES THAT MAKE
UP THE LUNG AND THE DIFFERENCES
BETWEEN HUMAN AND MOUSE LUNG AND
GENETICALLY WERE VERY WELL MADE
IN THIS PAPER PUBLISHED BACK IN
2020 IN WHICH THEY LOOKED AT
SINGLE CELL RES LAITION OF THE
GENE EXPRESSION PROFILE OF THE
CELLS THAT MAKE UP THE LUNG AND
FOUND MINORITY OF GENES ARE
ACTUALLY FAITHFULLY
RECAPITULATED IN TERMS OF THEIR
GENE EXPRESSION PATTERN IN THE
MOUSE LUNG.
FURTHERMORE, WHEN THEY LOOKED AT
INDIVIDUAL GENES IDENTIFY BY
GENOME WIDE ASSOCIATION STUDIES
WITH RELEVANCE TO CHRONIC LUNG
DISEASES, I'M SHOWING YOU DATA
FROM ONE OF THEM HERE, BUT
SIMILAR OBSERVATIONS WERE MADE
FOR OTHER GWAS GENES RELATED TO
COPD AND EMPHYSEMA SUCH AS HHIP,
THE GENE EXPRESSION PROFILE
DIFFERED IN WAYS NA COULD LIKELY
BE BIOLOGICALLY SIGNIFICANT.
SO THE POINT HERE IS SIMPLY THAT
WE NEED TO BE VERY CAREFUL,
THOUGHTFUL AND INTENTIONAL WHEN
WE'RE DESIGNING EXPERIMENTS TO
ADDRESS EXPERIMENTAL QUESTIONS
TO BE COGNIZANT OF THE RELATIVE
STRENGTHS AND WEAKNESSES OF THE
VARIOUS MODELS WE APPLY TO ASK
SPECIFIC QUESTIONS.
THE SECOND OBSERVATION THAT I
WOULD LIKE TO HIGHLIGHT THAT WE
ALL KNOW TO BE TRUE BUT MAYBE
DON'T THINK AS MUCH ABOUT IS
THAT COPD SUSCEPTIBILITY IS, IN
FACT, HERITABLE.
SO ONLY AROUND 20% OF LONG TERM
SMOKERS DEVELOP CLINICALLY
SIGNIFICANT COPD OR EMPHYSEMA
AND WE KNOW EITHER FROM GENETIC
STUDIES OR -- THIS IS REALLY A
GENETIC DISEASE AND WE KNOW THAT
MANY INDIVIDUALS IN THE
POPULATION DO NOT HARBOR THE
SUSCEPTIBILITY GENES THAT MAKE
THEM SUSCEPTIBLE TO THE SAME
INJURY THAT OTHERS ARE TERRIBLY
SUSCEPTIBLE TO AND I THINK THIS
IS A REALLY IMPORTANT POINT THAT
POTENTIALLY COULD BE A KEY TO
UNDERSTANDING DISEASE
PATHOGENESIS.
THEN LASTLY, I JUST WANT TO
HIGHLIGHT THE FACT THAT WE'VE
LEARNED SO MUCH AS WE ALL KNOW
FROM SINGLE CELL RNA SEQUENCING
STUDIES PROFILING DISEASED LUNG
FROM INDIVIDUALS WITH CHRONIC
LUNG DISEASES IN RECENT YEARS
AND THOSE INSIGHTS HAVE REALLY
BEEN TRANSFORMATIVE, BUT FOR
OBVIOUS REASONS, A LOT OF THE
DATA THAT HAS EMERGED HAS
FEATURED LUNGS THAT WERE SAMPLED
AT THE END STAGE OF THE DISEASE,
AND IT'S JUST IMPORTANT FOR US
TO BE AWARE OF THE FACT THAT
THAT DISEASE STATE MAY DIFFER
SIGNIFICANTLY FROM THE EVENTS
THAT ARE HAPPENING MOLECULARLY
IN THE EARLIEST STAGES OF THE
DISEASE, SO I THINK W IT WOULD E
WISE FOR US TO ALSO TRY TO THINK
CAREFULLY IN OUR FUTURE
EXPERIMENTS ABOUT UNDERSTANDING
THAT EARLY DISEASE STATE
PARTICULARLY IF OUR GOAL WAS TO
TRY TO DEVELOP NEW INTERVENTIONS
THAT CAN BE APPLIED BEFORE THE
LUNG HAS ADVANCED TO A VERY LATE
STAGE OF DISEASE.
AND SO WITH THESE OBSERVATIONS
IN MIND, MY LAB WAS THINKING
ABOUT HOW WE COULD APPLY MODEL
SYSTEMS TO UNDERSTAND DISEASE
PATHOGENESIS AND WE REALLY
WANTED TO APPLY A MODEL THAT
COULD FAITHFULLY RECAPITULATE
THE GENETICS OF THE HUMAN LUNG
THAT COULD CAPTURE THE
HETEROGENEITY OF HUMAN PATIENT
POPULATIONS AND THAT COULD BE
USED TO STUDY THE INCEPTION OF
DISEASE, THE EARLIEST DISEASE
STATE WHERE WE MIGHT BE ABLE TO
INTERVENE, AND SO WITH THOSE
THINGS IN OUR MIND, WE DECIDED
TO FOCUS ON HUMAN INDUCED
PLURIPOTENT STEM CELLS AND APPLY
THEM TO A MODEL OF COPD AND
EMPHYSEMA.
SO I WORK IN THE CENTER FOR
REGENERATIVE MEDICINE AT BOSTON
UNIVERSITY DI WITH DARRELL COTTN
AND THE DIFFERENTIATION OF
APPROACHES WE HAVE APPLIED HAVE
BEEN APPLIED BY HIS LAB AND
DEMONSTRATED IN THE LEFT-HAND
PANEL OF THIS SLIDE.
WE HAD A NICE DISCUSSION
YESTERDAY OF IPS DIRECTED
DIFFERENTIATION PROTOCOLS IN
PROTOCOL.OVER ABOUT A 35-DAY PEN
THIS CASE WE HAVE A SINGLE CELL
TYPE THAT'S REPRESENTED IN OUR
CULTURES AT THE END OF THE
PROTOCOL, AND WE CAN PURIFY
THOSE CELLS BASED ON THEIR
EXPRESSION OF MARKER GENES, IN
THIS CASE THERE'S A FLUORESCENT
REPORTER GENE KNOCKED INTO THE
SURFACTANT PROTEIN C LOCUS AND
WE CAN GENERATE A PURE
POPULATION CULTURED IN MATRIGEL
AND CAN BE SERIALLY PASSAGED FOR
ABOUT A YEAR.
COMPARING THEM TO PRIMARY CELL
TYPES AS IT WAS ILLUSTRATED IN
THIS FIGURE FR FROM THE STEM CEL
PAPER PUBLISHED IN 2017
DEMONSTRATING THIS CELLULAR
EXPRESSION PROFILE OF KEY TYPE T
OF PRIMARY CELLS.
THEN LASTLY IN THE PANEL OVER ON
THE RIGHT WHAT IS BEING
ILLUSTRATED HERE IS WHAT IA
T2s NND LAMELLAR BODIES IN
THOSE CELLS LOOK LIKE COMPARED
TO THE SAME CELLS BUT AFTER
CORRECTION OF THAT SINGLE
MUTATION AND YOU CAN SEE THAT
THE LAMELLAR BODIES IN THE TOP
PANEL ARE MALFORMED, HAVE AN
ABNORMAL APPEARANCE AND THEIR
APPEARANCE IS RESTORED TO A
NORMAL PATTERN FOLLOWING GENETIC
CORRECTION.
THE POINT HERE IS TWOFOLD, FIRST
THAT THESE CELLS CAN BE USED TO
MODEL FEATURES OF MONOGENIC
DISEASES BUT I THINK YOU CAN
EXTRAPOLATE THAT CONCEPT TO HOW
WE THINK OF THE CELLS AS
ESSENTIALLY BEING GENETIC
SURROGATES FOR THE PATIENT FROM
WHOM THEY WERE DERIVED AND IN
THIS CASE THEY'RE BEING APPLIED
TO UNDERSTAND THE CONSEQUENCES
OF A MUTATION IN A SINGLE GENE
BECAUSE THEY CONTAIN THE
PATIENT'S ENTIRE GENOME.
THEORETICALLY THEY CONTAIN ALL
OF THE VARIANTS THAT AN
INDIVIDUAL MIGHT HAVE THROUGHOUT
THEIR GENOME AND CAN, THEREFORE,
BE APPLIED TO UNDERSTAND THESE
COMPLEX DISEASES AS WELL.
SO WE KNOW OF COURSE THAT LIKE
OTHER DISEASES, COPD AND
EMPHYSEMA RESULT FROM A GENE BY
ENVIRONMENT INTERACTION.
MY LAB HAS TRIED TO DEVELOP
APPROACHES TO UNDERSTAND BOTH
THE GENETICS AND THE
ENVIRONMENTAL CONSEQUENCES THAT
ARE ASSOCIATED WITH THIS
DISEASE.
IMPLICIT IN OUR DECISION TO
FOCUS ON TYPE 2, WE ARE GOING TO
BE THINKING ABOUT EMPHYSEMA AS
OUR OUTCOME IN THIS PARTICULAR
SET OF STUDIES.
WHILE WE TYPICALLY SEE
CLINICALLY BOTH COPD AND
EMPHYSEMA WITHIN THE SAME
INDIVIDUALS BECAUSE THEY'RE ALL
EXPOSED TO THE SAME
ENVIRONMENTAL INJURY, THAT THOSE
ARE, IN FACT, SEPARATE OUTCOMES
THAT ARE DISTINCT TO THE
COMPARTMENTS THAT ARE AFFECTED
AND THOSE COMPARTMENTS OF COURSE
ARE MADE UP BY DIFFERENT CELL
TYPES THAT RESPOND DIFFERENTLY
TO INJURY.
BUT FOR THESE EXPERIMENTS, WE'RE
THINKING ABOUT SPECIFICALLY
EMPHYSEMA.
SO IN ORDER TO EXPOSE IA T2s
TO ENVIRONMENTAL INJURY, WE HAD
TO FIRST EXPOSE THEM TO THE
OUTSIDE STRIERMT ENVIRONMENT SOE
ADAPTED THE 3D PROTOCOL I
INTRODUCED YOU TO PREVIOUSLY TO
ALLOW THESE CELLS TO BE CULTURED
IN AN AIR LIQUID INTERFACE AS
ILLUSTRATED IN THE SCHEMATIC
HERE.
ONCE WE HAD DONE SO, WE FIRST
VALIDATED THE FACT THAT THESE
CELLS WERE MAINTAINING THEIR
IDENTITY AS TYPE 2 CELLS AND WE
DID SO BY LOOKING AT CONTINUED
EXPRESSION OF THE SURFACTANT
PROTEIN RECORDER GENE YOU SEE IN
THE BOTTOM PANEL RELATIVE TO THE
CELLS IN THEIR STANDARD 3D
CONDITIONS ON THE TOP.
BY DOING SO WE'RE ABLE TO
CONFIRM THESE CELLS MAINTAINED A
TYPE 2 CELL IDENTITY WHEN
CULTURED IN THE IDENTICAL
CONDITIONS.
WE WENT ON TO LOOK AT THE GENOME
WIDE EXPRESSION PROFILE OF CELLS
CULTURED IN THESE A.I.
CONDITIONS RELATIVE TO THE
STANDARD 3D CULTURE CONDITIONS
OR TO CELLS CULTURES IN A
TRANSWELL LABELED AS 2D, AND
WHAT WE FOUND WAS THAT SIMILAR
TO WHAT HAS BEEN PUBLISHED, THEY
IN FACT ENHANCE THEIR MATURATION
STATE AS READ OUT BY A PANEL OF
GENES WE NO ARE ASSOCIATED WITH
MATURATION OF TYPE 2 CELLS IN
PRIMARY HUMAN SAMPLES AS WELL AS
BY THE UPREGULATION OF A MODULE
OF GENES ASSOCIATED WITH
LAMELLAR BODY BIOGEN CYST.
SO HAVING ESTABLISHED THIS
CULTURE PLATFORM, WE WERE NEXT
ABLE TO APPLY TO TRY TO
UNDERSTAND THE CONSEQUENCES
TRANSCRIPTIONALLY OF CIGARETTE
SMOKE EXPOSURE AND TO DO SO WE
INVESTED IN THE TREUMENT YOU SEE
PICTURED HERE.
I THINK IF YOU LOOK CAREFULLY,
YOU CAN SEE THERE'S A CIGARETTE
IN THE EXPOSURE SLOT OF THE
UPPER LEFT-HAND -- OR OF THE
INSTRUMENT THERE, AND IF YOU
COULD CLICK ON THE PICTURE,
PLEASE, WE'LL BE ABLE TO SEE A
MOVIE OF THE SMOKING HAPPENING
IN ACTION, AND TH IT PASSES OVER
THE APICAL SURFACE OF THE CELLS
THAT ARE CULTURED IN AN
AIR-LIQUID INTERFACE AND THERE
ARE CONTROL CELLS EXPOSED TO AIR
IN THE SAME EXPERIMENT AND AFTER
A PERIOD OF TIME, WE CAN SAMPLE
THESE CELLS TO UNDERSTAND
TRANSCRIPTIONALLY WHAT IS
PERTURBED AS A CONSEQUENCE OF
CIGARETTE SMOKE EXPOSURE.
WHEN WE DO THAT, WE CAN GENERATE
THE DATA THAT YOU SEE HERE AND
THIS IS A SPRING PLOT SHOWING
SINGLE CELL RNA SEQUENCING DATA
FROM THESE CELLS IN FOUR
DIFFERENT CONDITIONS, THEY'RE
COLORED DIFFERENTLY BASED ON
THEIR CONDITION AND THEY'RE
LABELED TO IDENTIFY CELLS THAT
WERE CULTURED JUST IN 3D AND
SUBMERGED CULTURE AS WE
TYPICALLY DO, CELLS THAT WERE
CULTURED IN A TRANS WELL BUT
SUBMERGED IN CULTURE THAT WE
LABELED HERE IS 2D, CELLS THAT
WERE CULTURED AT AIR LIQUID
INTERFACE BUT EXPOSED TO AIR AS
CONTROLS THAT ARE LABELED AS
A.I. AND THEN THE -- THEN WE CAN
IDENTIFY DIFFERENTIALLY
EXPRESSED GENES COMPARING THE
SMOKED CELLS TO THE AIR CONTROLS
AND THEN LOOK AT PATHWAYS THAT
WERE UPREGULATED OR
DOWNREGULATED AS A CONSEQUENCE
BY GENE SET ENRICHMENT ANALYSIS.
I'M JUST SHOWING YOU THE
HALLMARK DATA FOR THE
UPREGULATED PATHWAYS HERE.
SO THEN WE COULD USE THIS DATA
TO TRY TO UNDERSTAND THE
CONSEQUENCES OF ACUTE SMOKE
EXPOSURE IN THIS PARTICULAR CELL
TYPE THAT CAN BE HARD TO ACCESS
FROM PATIENTS.
OF COURSE IN ADDITION TO
UNDERSTAND THE ENVIRONMENTAL
EXPOSURE WE WANT TO BE ABLE TO
UNDERSTAND THE GENE EXPRESSION
AND THE CONSEQUENCES OF
PERTURBATION -- GENOME WIDE
ASSOCIATION STUDIES TO BE
ASSOCIATED WITH COPD AND
EMPHYSEMA AND TO DO THIS WORK WE
TEAMED UP WITH THE CHANNING LAB
AND WE THOUGHT WITH THEM ABOUT
GENES IDENTIFIED BY GWAS THAT WE
THOUGHT WOULD BE LIKELY TO BE
BIOLOGICALLY RELEVANT IN TIP
2 CELLS AND THAT WERE EXPRESSED
IN HUMAN PRIMARY TIP 2 CELLS AS
WELL AS IN OUR IPSC PLATFORM.
AND TO TRY TO UNDERSTAND THE
FUNCTIONAL CONSEQUENCES OF THESE
GENES BOTH IN TERMS OF
HOMEOSTASIS AND ALSO IN TERMS OF
HAHOW THEY MIGHT CONTRIBUTE TO
DISEASE SUSCEPTIBILITY OR
PATHOGENESIS, WE MODIFIED OUR
IPSC PLATFORM TO USE AN APPROACH
CALLED CRISPR INTERFERENCE, AND
SO TO DO THAT, WE KNOCKED A
DCAS9 CRAB CONSTRUCT INTO THE
AAVS1 HARBOR LOCUS OF OUR CELLS
AND TOOK THEM TO THE NORMAL
DIFFERENTIATION PROTOCOL TO
GENERATE IA T2s AND INTRODUCED
GUIDE RNAs TO KNOCK DOWN THE
SPECIFIC GENES OF INTEREST IN A
LENTIVIRUS AND THEN WE COULD
SORT CELLS THAT WERE TRANSDUCED
WITH THIS BFP REPORTER TO
GENERATE PURE POPULATIONS OF IA
T2s EXPRESSING THE GUIDE
RNAs.
THIS IS -- WE WERE ABLE TO
TEMPORALLY CONTROL KNOCKDOWN BY
THE INTRODUCTION OF DOXYCYCLINE
TO THE TISSUE CULTURE MEDIA AND
THEN WE APPLIED THIS TO THE
APPROACH TO THAT NINE GENES WE
HAD SELECTED.
TO KNOCK THOSE GENES DOWN ONE AT
A TIMEMENT I'M SHOWING YOU THE
QPCR DATA FROM THAT EXPERIMENT
HERE, SHOWING WE CAN LITERALLY
KNOCK DOWN THESE GENES OF
INTEREST FROM GWAS.
SO BECAUSE TYPE 2 CELLS ARE
PROGENITOR CELLS IN THE AL VEE
ALVEOLUS, WE THOUGHT IT WOULD BE
USEFUL TO LOOK AT CELL FUNCTION
AND THE EFFECT OF KNOCKING DOWN
THOSE GENES ON PROGENITOR CELL
FUNCTION.
I'M NOT SHOWING YOU THE DATA
HERE BUT WE LOOKED AT PHENOTYPES
THAT WE THOUGHT WERE RELEVANT
FOR PROGENITOR CELLS THAT
INCLUDED PROLIFERATION,
MATURATION AND DIFFERENTIATION
EFFICIENCY AND THEN FROM THAT
PANEL OF NINE GENES, WE SELECTED
ONE TO FOCUS ON AND DO A DEEP
DIVE ON AND THAT GENE WAS DSP,
AN INTERESTING GENE IN PART
BECAUSE IT'S BEEN IDENTIFIED
INDEPENDENTLY BY GENOME WIDE
ASSOCIATION STUDIES TO BE
ASSOCIATED WITH BOTH COPD BUT
ALSO WITH PULMONARY FIBROSIS.
THIS IS SORT OF AN OVERVIEW OF
THE TYPE OF DATA THAT WE WERE
ABLE TO GENERATE IN THESE
STUDIES SO WE COULD LOOK
TRANSCRIPTIONALLY BY SINGLE CELL
RNA SEQUENCING ON THE LEFT AT
THE GENE EXPRESSION PROFILE OF
CELLS WITH DSP KNOCKDOWN
COMPARED TO CONTROLS WITHOUT
KNOCKDOWN.
AND THEN FROM THAT GENE
EXPRESSION DATA, WE COULD
IDENTIFY MOLECULAR PROCESSES
THAT WERE DYSREGULATED, IN THIS
CASE METABOLISM, AND I'M SHOWING
YOU A DATA FROM A SEAHORSE ASSAY
TO INTERROGATE MITOCHONDRIAL
FUNCTION IN THE CONTEXT OF DSP
KNOCKDOWN AND WE LIKEWISE
PERFORMED COMPLEMENTARY
EXPERIMENTS TO OVEREXPRESS DSP
USING AN APPROACH CALLED CRISPR
ACTIVATION TO SEE THE RESULTS OF
THAT WHICH IN THIS CASE WERE
EXACTLY OPPOSITE OF WHAT YOU'RE
SEEING HERE.
THEN LASTLY, WE COULD
INCORPORATE THE CIGARETTE SMOKE
INJURY APPROACH THAT I
INTRODUCED TO YOU PREVIOUSLY TO
INTERROGATE GENE BY ENVIRONMENT
INTERACTIONS AND UNDERSTAND IF
THEY HAVE CONSEQUENCE OF THAT ON
PROGENITOR CELL FUNCTION BY
COLONY FORMING EFFICIENCY.
WHILE I'VE JUST SHOWN YOU ALL OF
THIS DATA AS IT APPLIES TO A
SINGLE GENE AND A CELL TYPE, I
THINK THIS IS A GENERALIZABLE
APPROACH THAT COULD BE USED TO
UNDERSTAND THE HOMEOSTATIC AND
POTENTIALLY DISEASE RELEVANT
CONSEQUENCES OF PERTURBING ANY
GENE AND WHILE I'VE SHOWN TO YOU
IN IA T2s, IT COULD ALSO BE
PERFORMED IN AIRWAY CELLS AS
WELL.
WHEN I THINK ABOUT CURRENT GAPS
IN UNDERSTANDING ASSOCIATED WITH
THIS TYPE OF DATA, I THINK THAT
IT'S IMPORTANT TO THINK ABOUT
THE GENETIC CONTRIBUTIONS TO
SUSCEPTIBILITY AND PATHOGENESIS
TO UNDERSTAND BETTER THE
HETEROGENEITY THAT EXISTS WITHIN
PATIENTS WHO HAVE COPD AND
EMPHYSEMA AND ALSO THE
ASSOCIATED MOLECULAR PHENOTYPING
DIFFERENCES THAT ARE ASSOCIATED
WITH THOSE DIFFERENT PHENOTYPES.
TO THINK ABOUT THE CELL TYPE AND
COMPARTMENT SPECIFIC RESPONSES
TO DISEASE RELEVANT INJURY AND
AS I'VE MENTIONED ALREADY TO
FOCUS ON PATHOGENESIS OF EARLY
STAGE DISEASE.
THEN LASTLY, SOME KEY RESEARCH
OPPORTUNITIES THAT I THINK WOULD
BE VALUABLE TO THINK ABOUT WOULD
BE TO IDENTIFY LONGITUDAL COHORT
STUDIES THAT HAVE ASSOCIATED
BIOREPOSITORIES THAT WOULD ALLOW
US TO LINK PRIMARY CELLS AND
TISSUES WITH LONGITUDAL
PHENOTYPING DATA.
SECONDLY, TO THINK ABOUT
EXPERIMENTAL VALIDATION OF GWAS
VARIANT FUNCTION IN
DISEASE-RELEVANT CELLS AND
LASTLY TO FOCUS ON HUMAN MODEL
SYSTEMS THAT CAN BE COUPLED
TOGETHER WITH EITHER PCLS OR
WITH ANIMAL MODELS THAT
RECAPITULATE ESSENTIAL FEATURES
OF THA HUMAN LUNG STRUCTURE.
THANK YOU VUCH.
VVERY MUCH.
I'D BE HAPPY TO TAKE ANY
QUESTIONS AT THE END.
>> QUESTIONS AT THE END.
I'M HAROLD CHAPMAN.
I JUST WANT TO INTRODUCE THE
NEXT SPEAKER.
THESE ARE ALL -- IN THIS
SESSION, VERY HUMAN-ORIENTED
PRESENTATIONS, AND THE ONE
THAT'S COMING UP IS BY DAN
WEISS.
HE'S A PROFESSOR OF MEDICINE IN
BIOENGINEERING AT THE UNIVERSITY
OF VERMONT.
HE'S BEEN INTERESTED IN THE
POTENTIAL THERAPEUTIC POTENTIAL
OF MESENCHYMAL STEM CELLS FOR A
LONG TIME, AND I THINK HE'S
GOING TO UPDATE US ON WHERE THIS
FIELD STANDS.
DAN?
>> ALL RIGHT, THANK YOU VERY
MUCH, HAROLD, AND THANKS TO THE
ORGANIZERS FOR INCLUDING THIS IN
THE OVERALL SYMPOSIUM.
THIS IS A LITTLE BIT DIFFERENT
FROM ALL OF THE WORK YOU'VE SEEN
SO FAR BUT I WANT TO HIGHLIGHT
THE POTENTIAL OF CELL-BASED
THERAPY USE CELLS THAT DON'T
NECESSARILY GRAFT.
SO WHAT WE'LL DO IS WE'LL GO
OVER A LITTLE BIT OF THE
RATIONALE BACKGROUND ON WHAT
THESE MSCs ARE, KNOWN
MECHANISMS OF ACTION AND A
LITTLE BIT OF PRE-CLINICAL
STUDIES.
BUT WHAT I WANT TO DO IS FOCUS
ON THE CLINICAL INVESTIGATIONS
TO DATE IN COPD AND ONE OF THE
THINGS WE'RE GOING TO HIGHLIGHT
IS WHAT'S BEEN BROUGHT UP SO FAR
THAT COPD LIKE SO MANY OTHER
DISEASES IS A HETEROGENEOUS
SYNDROME, AND SO IT'S NOT GOING
TO BE ONE SIZE FITS ALL.
AND WE'LL TALK A BIT ABOUT
NON-COVID AND COVID RDS
APPLICATIONS.
BUT THE KEY THING IS, THERE'S
BEEN A LOT OF WORK AND SOME OF
THESE TRIALS WORK, SOME OF THEM
DON'T, AND WE NEED TO UNDERSTAND
BETTER ABOUT HOW WE CAN MAKE
THEM ALL WORK.
SO A LITTLE ABOUT MY LAB
INTERESTS AND ALL THAT.
WE'RE GOING TO TALK ABOUT THE
UPPER RIGHT-HAND CORNER
CELL-BASED THERAPIES BUT I WANT
TO POINT OUT, WE'VE DONE A LOT
OF WORK IN THE LOWER RIGHT-HAND
CORNER THERE WITH HYDROGELS AND
DEVELOPED A SERIES OF TOOLS WITH
LUNG EXTRACELLULAR MATRIX
DERIVED HYDROGELS FROM NORMAL
AND DISEASED LUNGS, COPD, AND
IPF THAT I THINK COULD BE
VALUABLE ADDITIONS TO A LOT OF
THE REALLY GREAT WORK THAT'S
GONE ON SO FAR.
SO WHAT ARE THESE M SCs?
THESE ARE CELLS ORIGINALLY
ISOLATED FROM THE BONE MARROW
AND NOW HAVE BEEN FOUND TO BE
PRESENT IN A RANGE OF TISSUES
INCLUDING ADIPOSE, PLACENTAL,
CORD BLOOD AND OTHERS.
THEY DON'T LEAVE THESE TISSUES,
THEY HAVE TO BE EXTRACTED AND
FOR A VARIETY OF IN VITRO
SYSTEMS, WHAT ONE CAN DO IS
INCUBATE THE CELLS WITH A RANGE
OF IMMUNE EFFECTOR CELLS
INCLUDING T CELLS, B CELLS, ET
CETERA, AS LISTED, AND IN
VIRTUALLY EVERY CASE, THE
PRESENCE OF THE MSC WILL INHIBIT
BOTH PROLIFERATION AND THE
PARTICULAR IMMUNE EFFECTOR
FUNCTION OF THOSE CELLS.
THE SCHEMATIC ON THE RIGHT SIDE
SHOWS -- IT'S AN OLD SCHEMATIC,
BUT RANGE OF SECRETED MEDIATORS
THAT HAVE BEEN IMPLICATED IN
THESE ACTIONS.
THIS INCLUDES ALSO EXTRACELLULAR
VESICLES AND THE EXTRACELLULAR
VESICLE CONTENT.
THE MSCs ALSO HAVE POOR
INITIAL IMMUNOGENICITY WHICH
MEANS ONE CAN UTILIZE NON-H LA.
MATCHED ALLOGENEIC MSCs AND
CLINICAL SCHEMES, IF YOU WILL.
IN TERMS OF CLINICAL
APPLICATIONS, THERE'S BEEN A
WIDE RANGE OF INVESTIGATIONS
BASED ON SOLID PRE-CLINICAL DATA
IN DISEASES THAT ARE MOSTLY
AUTOIMMUNE AND INFLAMMATORY AS
LISTED, GRAFT-VERSUS-HOST
DISEASE, CROHN'S, MULTIPLE
SCLEROSIS, TYPE 1 AUTOIMMUNE
DIABETES, ET CETERA.
CARDIAC DISEASE AND OTHERS.
AND TO DATE, THE MSC BASED
CLINICAL THERAPIES ARE APPROVED
FOR A LIMITED RANGE OF DISEASES.
REFRACTORY PEDIATRIC GRAFT
SWGRAFT-VERSUS-HOST DISEASE, MOT
RECENTLY THIS FIRST IN THE U.S.
ORPHAN DRUG DESIGNATION FOR
HYPOPLASTIC LEFT HEART SYNDROME.
IN THE WORLD OF -- WELL,
ACTUALLY SO HOW DOES THIS WORK?
SO AGAIN, THE CELLS DON'T LEAVE
THEIR SOURCE OF ORIGIN, AND WHAT
ONE HAS TO DO IS HARVEST THEM,
EXPAND IN CULTURE AND THERE ARE
MULTIPLE APPROACHES TO DOING
THAT, AND THEN RE-ADMINISTER, OR
ADMINISTER.
MOSTLY SYSTEMICALLY, BUT FOR
LUNG APPLICATIONS, THERE IS A
RICH PRE-CLINICAL LITERATURE ON
DIRECT INTRATRACH YE INTRATRACHY
DELIVERY.
WHAT HAPPENS IN SYSTEMIC
ADMINISTRATION IS THAT THE CELLS
WILL LODGE IN THE LUNGS MOSTLY
ON THE FIRST PASS.
THEY DON'T ENGRAFT.
THIS IS CRITICALLY IMPORTANT.
THEY SIT THERE FOR A FEW DAYS,
THEY INTERACT WITH VARIOUS
INTEGRINS ON THE CELL SURFACE
AND THEY EXPRESS TOLL LIKE
RECEPTORS AND OTHER DAMAGED
ELECTROPATHOGEN ASSOCIATED
RECEPTORS SUCH THAT THEY WILL
RESPOND TO AND REACT ACCORDINGLY
BY RELEASE OF A PORTFOLIO OF
SUBSTANCES INCLUDING
ANTI-INFLAMMATORY MOLECULES TO
ANY GIVEN INFLAMMATORY
SITUATION, AND THEN THEY'RE
CLEAR, AND CLEARANCE MECHANISMS
INCLUDE PHAGOCYTOSIS BY RESIDENT
MACROPHAGES AND OTHERS.
SO THESE CELLS ARE SHORT LIVED
BUT DO HAVE A VERY POTENT
EFFECT.
SO IN THE WORLD OF LUNG AND
RESPIRATORY ILLNESSES, THERE IS
A VERY LARGE LITERATURE, OVER AT
LEAST 500 PUBLICATIONS IN A
VARIETY OF PRE-CLINICAL, THIS
INCLUDES BOTH RODENT AS WELL AS
LARGER ANIMALS SUCH AS PIGS,
SHEEP AND ALL, THAT ONE CAN
INDUCE A VARIETY OF DIFFERENT
TYPES OF LUNG INJURIES.
YOU CAN SEE IN THE RIGHT HAND
GOING AROUND THE CIRCLE, THE
RANGE OF DIFFERENT INJURY MODELS
UTILIZED, AND IN VIRTUALLY EVERY
CASE, SYSTEMIC ADMINISTRATION
AND IN MANY CASES DIRECT
INTRATRACHEAL ADMINISTRATION OF
THE CELLS WILL AMELIORATE
WHATEVER YOUR PARTICULAR
INFLAMMATORY OR INJURY END
POINTS ARE.
AND AGAIN, THERE ARE A VARIETY
OF POSTULATED MECHANISMS
INCLUDING CELL CONTACT
INDEPENDENT SOLUBLE MEDIATORS,
INCLUDING A VARIETY OF
CYTOKINES, ET CETERA, THE
EXTRACELLULAR VESICLES, THERE'S
SOME DEGREE POTENTIALLY OF CELL
CONTACT DEPENDENT IN WHICH
MITOCHONDRIA ARE TRANSFERRED
THROUGH CONNECTION 43 BRIDGES
FROM THE MSCs TO THE RESIP
YEBT CELLS AND THERE'S SOME
INDIRECT MECHANISMS.
FOR EXAMPLE, WE'VE DONE A LOT OF
WORK IN PRE-CLINICAL MODELS WITH
ASTHMA AND THE MSCs ACTUALLY
DON'T NECESSARILY DIRECTLY
AFFECT THE LUNG BUT RATHER
AFFECT THE I MU -- CELLS SO THIS
HAS BEEN APPLIED INCREASINGLY SO
OVER THE PAST NUMBER OF YEARS TO
A VARIETY OF CLINICAL
INVESTIGATIONS, AND TO GIVE YOU
THE LANDSCAPE, MOST OF THESE ARE
PHASE ONE SAFETY.
DOSE ESCALATION.
THERE'S A LIMITED BUT GROWING
NUMBER OF PHASE 2 EFFICACY
TRAILS BUT THERE IS STILL NO
CONSENSUS ON APPROACH, AND THIS
INCLUDES, WHERE DO YOU GET THE
MSCs FROM?
BONE MARROW, ADIPOSE, PLACENTAL,
ET CETERA.
DO YOU USE AUTOLOGOUS,
ALLOGENEIC, AND THERE ARE PROS
AND CONS FOR BOTH.
THE MANUFACTURING, IE, HOW YOU
ISOLATE THE CELLS, HOW YOU
EXPAND THEM IN TISSUE CULTURE IS
A WIDE OPEN FIELD, IF YOU WILL.
CELLS WERE MOSTLY UNMANIPULATED.
ONE CAN ENGINEER, FOR EXAMPLE,
MSC TO EXPRESS ANTITUMOR
MOLECULES, FOR EXAMPLE, TRAIL,
APOPTOSIS, INDUCING LIGAND AND
THERE HAVE BEEN INTRIGUING
STUDIES IN DISEASES SUCH AS
MESOTHELIOMA AND LUNG CANCER
UTILIZING THESE.
THERE IS NO SET DOSE OR DOSING
REGIMEN.
WEEWE'LL COME BACK TO THAT IN A
MINUTE.
WE GOT INTO THIS A NUMBER OF
YEARS AGO WORKING WITH A COMPANY
AND I HAVE NO DISCLOSURE HERE, I
WAS A P.I. FOR THIS COMPANY
TRIAL, AND THIS WAS FOR COPD.
IF YOU THINK ABOUT IT, COPD MAY
NOT NECESSARILY BE THE BEST
DISEASE TARGET FOR MSC-BASED
CELL THERAPY AND THE RATIONALE
HERE IS THAT THE MSCs WILL BE
ADMINISTERED, STICK AROUND FOR A
FEW DAYS AND THEN THEY'RE
CLEARED.
SO FOR ACUTE HIGH INTENSITY
INFLAMMATORY SITUATION LIKE ARDS
OR SEPTIC SHOCK, THIS WOULD BE A
VERY GOOD STRATEGY.
IT'S MORE ON THE TABLE AS TO
WHETHER OR NOT A CHRONIC
INFLAMMATORY SITUATION OR
DESTRUCTIVE DISEASE LIKE COPD
WOULD BE AMENABLE, BUT
NONETHELESS, THIS IS AN
OPPORTUNITY TO DO AN INITIAL
SAFETY TRIAL IF NOTHING ELSE, SO
THIS IS A PHASE TWO MULTICENTER
RANDOMIZED DOUBLE BLIND
CONTROLLED -- THE PLACEBO WAS
THE VEHICLE -- NOT A DIFFERENT
CELL AND YOU CAN SEE IN THE
DIAGRAM, 65 PATIENTS, SIX SITES,
FOUR INFUSIONS OVER THE COURSE
OF FOUR MONTHS.
THERE WAS NO RIGHT OR WRONG TO
THIS, IT WAS JUST A STARTING
POINT.
SECONDARY OUTCOMES WERE
PULMONARY FUNCTION, QUALITY OF
LIFE, ET CETERA, BUT WE WERE
ALSO ABLE TO BUILD IN AN
INVESTIGATION TALL ARM IN WHICH
WE LOOKED AT CIRCULATING
INFLAMMATORY MEDIATORS.
SO THE OUTCOME WAS THAT THERE
WERE NO INFUSIONAL TOXICITIES,
NO SIGNS, SYMPTOMS OF CELL
EMBOLI, PULMONARY EMBOLI, NO
SERIOUS ADVERSE EVENTS,
FOLLOW-UP USING THE OUTCOME
MEASURE AS LISTED.
THE STUDY AT 62 PATIENTS WARS
UNDERPOWERED TO ARE EFFICACY AND
THERE WAS NO EUREKA MOMENT, BUT
WE SAW NO CHANGE IN LUNG
VOLUMES, DLCO, SIX MINUTE WALK,
ET CETERA.
BUT WHAT WE DID FIND WAS THAT IN
PATIENTS WITH ELEVATED C
REACTIVE CIRCULATING PROTEIN, AT
STUDY ONSET THERE'S
STATISTICALLY SIGNIFICANT
DECREASE IN THE PATIENTS
RECEIVING THE CELLS AS OPPOSED
TO PLACEBO AND OVER THE COURSE
OF THE INFUSION PERIOD AND IT
REMAINED AS A TREND OVER THE
TWO-YEAR OBSERVATION PERIOD.
SO WE SAT ON THIS FOR A LONG
TIME AND THEN JUST REALIZED AS A
GROWING APPRECIATION ABOUT THE
HETEROGENEITY OF COPD -- WE WENT
BACK AND TOOK A LOOK AS TO
WHETHER OR NOT WE COULD STRATIFY
PATIENTS BASED ON INFLAMMATORY
PHENOTYPE.
SO WE WENT BACK AND DID A
RETROACTIVE STUDY FOCUSING ON
THE CRP.
CRP, WIDELY RECOGNIZED MARKER OF
INFLAMMATION IN COPD PATIENTS,
HIGHER MORTALITY, WORSE
EXACERBATION OUTCOMES, HIGHER
READMISSION RATE, ET CETERA.
SO WE SIMPLY JUST RESTRATIFIED
THE PATIENTS BASED ON LEVELS OF
CIRCULATING CRP STUDY ENTRY AS
LISTED.
WHAT WE SAW WAS THAT IN THE
PATIENTS IN THE HIGHEST QUARTILE
OF CRP GREATER THAN 4, THERE IS
BOTH STATISTICALLY SIGNIFICANT
AS WELL AS CLINICALLY MEANINGFUL
IMPROVEMENT IN OUTCOMES,
SPECIFICALLY FEV1, FEC, AND
6 MINUTE WALK DISTANCE AND YOU
CAN SEE IN THE GRAPHS OVER ON
THE RIGHT THAT THE TREND FOR
FEV1, 6 MINUTE WALK DISTANCE
LASTED OVER THE TWO-YEAR STUDY
PERIOD.
THIS IS OF COURSE WITH ALL THE
CAVEATS OF A RETROSPECTIVE POST
HOC LOOK AT A NUMBER OF --
LIMITED NUMBER OF PATIENTS BUT
IT OPENS DOOR THAT INFLAMMATORY
PHENOTYPE IS REALLY CRITICALLY
IMPORTANT WHEN YOU'RE TRYING TO
APPLY A CELL-BASED THERAPY.
SO THAT TAKING THIS OVER INTO TE
ACUTE INJURY WORLD WITH RESPECT
TO ARDS, AGAIN, THERE'S A FIRM
LARGE PRE-CLINICAL BACKGROUND IN
MODELS OF ACUTE LUNG INJURY OF
MSC ADMINISTRATION AS REALLY
QUITE EFFICACIOUS, AND THERE
WERE A FEW TRIALS IN NON-COVID
ARDS PRIOR TO THE PANDEMIC, BUT
ONCE THE PANDEMIC HIT, THERE WAS
A HUGE NUMBER WITH AT LEAST 75
DIFFERENT CLINICAL
INVESTIGATIONS LISTED ON
CLINICAL TRIAL.GOV AS WELL AS
THE WHO, WOR WORLD HEALTH
ORGANIZATION REGISTRY, AND THE
CHINESE CLINICAL TRIAL REGISTRY,
AND MOST OF THESE WERE PHASE ONE
UNBLINDED NON-RANDOMIZED AND ALL
THAT, BUT YOU CAN SEE HERE IN
THE SCHEMATICS WHICH ARE BASED
ON SOME WORK BY MICHAEL MATTAY
AND HIS GROUP, POTENTIAL
MECHANISMS OF MECHANISMS IN BOTH
COVID I!
SARS-COV-2 AND NON-COVID, SO A
LOT OF QUESTIONS TO PURSUE HERE.
THE SHORT STORY IS THAT SOME OF
THESE TRIALS WORKED.
THIS WAS A TRIAL FROM THE
UNIVERSITY OF MIAMI PUBLISHED IN
2021, AND IT WAS A DOUBLE BLIND
RANDOMIZED CONTROL TRIAL PLACEBO
HERE AND YOU CAN SEE THE DETAILS
OF THE TRIAL DIAGRAM HERE WAS
RELATIVELY SMALL, WITH 12
RECEIVING THE CELLS, 12
RECEIVING THE VEHICLE.
PRIMARY OUTCOMES WERE ADVERSE
EVENTS AND THE LIKE.
SHOWS THAT THIS TRIAL WORKED, SO
IF YOU LOOK AT THE SURVIVAL
CURVE ON THE LEFT, ADVERSE
EVENT, FREE TRIAL ON THE
RIGHT -- EXCUSE ME THE CENTER
AND THE TIME FOR RECOVERY.
THESE WERE ALL STATISTICALLY AND
CLINICALLY MEANINGFULLY IMPROVED
IN THESE PATIENTS.
SO SOME RECENT META-ANALYSES
SHOW THAT AMONGST THE DOUBLE
BLIND RANDOMIZED PLACEBO
CONTROLLED TRIALS, THERE'S A
CLEAR BENEFIT TO MSC
ADMINISTRATION.
THE QUESTION IS, WHY DON'T THEY
ALWAYS WORK?
SO TO PULL THIS ALL TOGETHER,
SOURCE MANUFACTURING DOSE DOSING
HAVE BEEN QUESTIONS FOR A LONG
TIME.
THE INFLAMMATORY -- OF THE
PATIENT IS REALLY WHERE I THINK
THE FOCUS SHOULD BE RIGHT NOW.
ONE OF THE THINGS WE'RE
SPEARHEADING, THIS IS THROUGH
THE INTERNATIONAL SOCIETY OF
CELL AND GENE THERAPY IS
COLLABORATIVE TRIAL WITH THE
INVESTIGATORS OF THOSE DOUBLE
BLIND RANDOMIZED PLACEBO
CONTROLLED TRIALS IN WHICH WE
WERE OBTAINING INDIVIDUAL
PATIENT DATA AND ASKING THE
QUESTION CO DOES THE INFLAMMATOY
PROFILE AS MEASURED BY
CIRCULATING INFLAMMATORY
MEDIATORS PREDICT OUTCOMES AS A
RESPONSE TO CELL-BASED THERAPY?
SO WE THINK THIS IS GOING TO
OPEN UP DOORS THAT WE'RE REALLY
FOCUSED MORE ON PERSONALIZED
MEDICINE IF YOU WANT TO USE THAT
TERM, BUT FINDING THE BEST
POPULATION MOST LIKELY TO
RESPOND.
SO WITH THAT, I'M GOING TO PUT
IN A PLUG, IF I MAY, FOR THE
CONFERENCE O, VERMONT STEM CELL
CONFERENCE THAT MANY OF YOU HAVE
BEEN TO.
NEXT JULY '25 IS OUR NEXT
ANNIVERSARY, INVITE EVERYBODY TO
PARTICIPATE IN THAT.
I WANT TO THANK, OF COURSE,
MANY, MANY, MANY PEOPLE I'VE HAD
THE PRIVILEGE OF BEING
ASSOCIATED WITH AND BEING ABLE
TO WORK WITH.
WITH THAT, THANK YOU, AND WE CAN
CONTINUE DISCUSSION IN THE
DISCUSSION PERIOD.
>> OKAY.
THANK YOU, DAN.
THE NEXT SPEAKER IS DIANHUA
JIANG, THAT'S MY BEST CHINESE,
SORRY, WHO IS GOING TO -- IS
FROM THE CEDARS-SINAI MEDICAL
CENTER IN L.A., AND HE'S BEEN
DEVELOPING A REALLY INTERESTING
STORY ON THE METABOLOMICS OF
STEM CELLS, AND A PARTICULAR
FOCUS IN, AS THE TITLE SAYS, ON
ZINC IN IPF, AND HE'S GOING TO
TAKE US WHERE HE THINK THAT'S
HEADED. SO TAKE IT AWAY,
DIANHUA.
>> THANK YOU.
GOOD MORNING, EVERYBODY.
YES.
FIRST OF ALL I'D LIKE TO THANK
THE ORGANIZERS FOR INVITING ME
TO PRESENT MY RECENT WORK ON
ZINC METABOLISM IN IPF.
IPF PATHOGENESIS, SEVERAL
SPEAKERS HAVE TOUCHED ON.
I JUST WOULD LIKE TO BRIEFLY
HIGHLIGHT THE ROLE OF THE
IMPORTANCE CONCEPTUALLY IN THE
LAST 15 YEARS THAT
DIFFERENTIATION OF -- FUNCTION
AS STEM CELLS IN THE DISTAL
AIRIZED SPACE.
FOR THIS TALK, I WOULD LIKE TO
FOCUS ON THE INTRINSIC A T2 CELL
WHICH MIGHT BE THE KEY DRIVERS
FOR IPF PATHOGENESIS.
IT HAS LONG BEEN SUGGESTED
THAT -- STEM CELL EXHAUSTION IN
HIV LUNG AND SEVERAIPF LUNG, WEE
CAN SEE THE SIGNIFICANT LOSS OF
CELLS FROM IPF LUNG.
THIS LEADS TO -- CULTURE SYSTEM
FROM -- COLONY.
AND WE FURTHER SHOW THAT THE
INTERACTION BETWEEN
EXTRACELLULAR MATRIX -- AND THE
TR4 CAP 2 CELLS -- AS
ILLUSTRATED BY THE DIAGRAM FRO
FROM -- CHAMBERS PAPER.
AFTER THAT STUDY, WE HAVE
STUDIES TO LACK INTO WHAT ARE
THE -- HI IPF AND LOSS OF A T2 M
CELLS.
WE STARTED LOOKING AT SOME OF
THE CELLULAR METABOLISM
DISREGULATIONS IN IPF -- CELLS.
FOR THIS TALK I'LL MOSTLY TALK
ON ZINC AND I'LL TOUCH A LITTLE
BIT ON -- PLUS PLUS AND A LITTLE
ON THE IPF METABOLISM.
WE DID SINGLE CELL RNA
SEQUENCING WITH FRESHLY ISOLA
ISOLATED -- CELLS FROM SIX
DONORS AND SIX HIV EXPLANTS, AND
WE CAN CLEARLY SEE THAT THE LOSS
OF -- CELLS -- CLUSTERS AND IN
STRIKING GENES LOST IN TYPE
2 CELLS CALLED THE -- 9AA AND
THIS GENE ENCODES A PROTEIN
CALLED THE ZINC A WHICH IS THE
ZINC -- YOU CAN SEE THE DIAGRAM
TO THE RIGHT AND THE -- CULTURE
BASICALLY TRANSPORT THE ZINC
FROM EXTRACELLULAR SPACE TO
THE -- ZINC CONCENTRATION INSIDE
THE CELL AND PERFORM BIOLOGICAL
FUNCTIONS.
I CAN CONFIRM THIS LOSS OF -- BY
RTP -- BY FLOW CYTOMETRY AND
ALSO CONFIRM FROM THE LGRC
DATASETS, AND THE LUNG TISSUE,
LOSS OF -- LUNG FUNCTION.
SO WITH TH THE SIGNIFICANT LOWER
ZINC EXPRESSION -- EXPECT THAT
THERE WOULD BE A SIGNIFICANT
LOWER ZINC INTRACELLULARLY AND
IF WE INDEED USE THE -- TO
MANAGE ZINC AND WE CAN SEE IN
PANEL A AND B AND WE SEE A
SIGNIFICANT LOWER ZINC -- IPF --
CELLS.
AND WE ALSO TAKE ADVANTAGE OF --
THE EXTRACELLULAR -- I MEAN THAT
IS ON THE CELL MEMBRANE AND --
HIGH AND LOW -- TYPE 2 CELLS WE
CAN CHARACTERIZE THAT -- TYPE
2 CELERY NEWELL CAPACITY IN
32 ORGANOID CULTURE SHOWING IN
PANEL D THAT -- LEAD TO --
COLONY WELL.
AND WHEN WE USE THAT -- ZINC IN
CULTURE, WE CAN SEE THE ZINC --
REMUABLE -- IPF TYPE 2 CELLS.
THAT'S SHOWING YOU PANEL F BU
BUT -- SHOWING IN PANEL G.
SO NEXT WE LACK INTO WHAT'S THE
MECHANISM RELATED TO THIS TYPE .
2 CELL RENEWAL.
WE DID PATHWAY ANALYSIS, WE
FOUND THAT -- PATHWAYS IS
SIGNIFICANT DOWN REGULATED IPF
TYPE 2 CELLS, AND IT'S
INTERESTING -- THEY REQUIRE SOME
ZINC FOR THEIR ACTIVITY, AND AT
LEAST THERE ARE LOTS OF STUDIES
LACKING INTO THE -- AND STEM
CELL RENEWABLE AND ZINC IS THE
REQUIRED ELEMENT.
WE CAN SEE IN PANEL B, WE CAN
USE FRESH ISOLATE OF -- CELLS
CONFIRM THE THIRD ONE -- IN IPF
CELLS AND FURTHERMORE, IF WE
CAN -- IF WE USE SIRNG TREATMENT
AND WE CAN SEE -- TYPE 2 CELL
EXPRESS A CERTAIN ONE BUT NOT
MUCH IN IPF TYPE 2 CELLS PANEL
C, AND THEY ARE NOT -- SHOWN IN
PANEL D THERE H.
THERE HAVE BEEN LOTS OF
COMPOUNDS RELATED TO --
MANIPULATE -- A CERTAIN ONE
ACTIVITY, AND THE ONE FOR -- WE
HAVE -- WE USE IN ACTIVATOR
SRT1720 IN PANEL E, AND WE SEE
THAT -- ACTIVATOR ENHANCE TYPE T
THOSE PROCESS.
INTERESTING ENOUGH, SHOWING IN
PANEL H, IT LOOKS LIKE A ZINC
TREATMENT AND ACTIVATOR -- TYPE
2 CELERY NEWABLE.
SO WE FURTHER DID TARGET
DIVISION OF TYPE 2 CELLS AND WE
SEE THAT -- WE CAN ISOLATE THOSE
CELLS AN MEASURE THEIR CELLULAR
ZINC SHOWING PANEL D,
SIGNIFICANT LOWER ZINC
INTRACELLULARLY AND THOSE CELLS
DO NOT -- WELL BUT IF WE FURTHER
INJURE THOSE -- ISOLATE TYPE
2 CELLS AND WE CAN CLEARLY SEE
THOSE -- WE HAVE MUCH LOWER TYPE
2 CELERY C2 CELL RECOVERY AND TS
DO NOT FORM COLONY WELL AND
SHOWING PANEL I AND J.
SO IF WE LEAVE THOSE MICE LON
LONGER -- LEAVE THEM ---YEA
THEM ---YEAR-OLD -- PANEL B AND
PANEL C, AND FURTHERMORE, IF WE
GIVE ONE YEAR -- MICE -- -- LUNG
TISSUE -- AND WE CAN CLEARLY SEE
THAT -- MICE THEY HAVE A LOWER
SURVIVAL AND MUCH HIGHER --
SHOWING YOU IN PANEL F AS THE
HYDROXY -- COMPOUND.
WE ALSO TAKE ADVANTAGE OF
ZINC -- AVAILABLE AND WE GIVE
THEM ZINC AND WE SEE IT CAN
REDUCE LUNG FIBROSIS SHOWING IN
PANEL H.
LOWER -- WORSE.
AS WE STARTED THIS, WE ALSO
NOTED THAT -- IF YOU LACK AT --K
AT SIRT1 IT REQUIRES ZINC AND
ALSO REQUIRES NAD PLUS, PRODUCED
BY NAMPT AND SO IN THE OLD MICE,
AND WE DO SEE -- REDUCE
EVENTUALLY IN THE -- MICE.
AND WE CAN CONFIRM THE NAMPT --
CELLS ARE FRESHLY ISOLATED TYPE
2 CELLS AND ALSO WE CAN --
LOOKING -- CONFIRM BY SINGLE
CELL -- IN PANEL C AND ALSO BY
IMMUNOFLUORESCENT STAINING AS
SHOWN IN PANEL D AND E.
BUT UNFORTUNATELY THERE'S LOTS
OF -- COMPOUNDS AVAILABLE AND --
AAT AND AT AND SIGNIFICANTLY EE
NAMPT ACTIVITY BUT WHEN WE
USE -- IN CULTURE AND --
SIGNIFICANTLY PROMOTES -- CELERY
NEWABLE AND SHOWING -- AS WELL
AS THE TYPE 2 -- -- THIS
PROCESS.
AND WE CAN ALSO TAKE ADVANT
ADVANTAGE -- ACTIVATION IN VIVO
AND WE TREAT THE MICE AND SAVING
THOSE AND HARVESTING -- FIVE
AND -- WE HAVE A MUCH BETTER
RECOVERY OF TYPE 2 CELLS, AND IF
WE LEAVE THEM 21 DAYS, WE CAN
SEE THAT -- SIGNIFICANTLY
REDUCED LUNG FIBROSIS.
SO THE LAST PIECE OF THIS
PRESENTATION, SO WE RECENTLY --
SUCH AS -- LIPIDS AFFECTING IPF
TYPE 2 CELLS, AND THE ONE WITH
THE GENES, THE LAST PANEL --
PANEL B AND THIS LPCAT1.
LPCAT1 LYSOPHOSPHATIDYLCHOLINE
ACYLTRANSFERASE 1, AND -- BODY
AND TO SECRETE EXTRACELLULAR
SPACE -- LPCAT 1 IS SPECIFICALLY
EXPRESSED IN TYPE 2 CELLS IN
LUNG TISSUE AND -- REDUCE THE
IPF TYPE 2 CELLS BY SINGLE CELL
RTPCR --.
WE GENERATED THE LPCAT1 DELETION
MICE, SO WE'RE LOOKING TO THOSE
CELLS AND -- ORGANELLE CULTURE.
THOSE TYPE 2 CELLS WITH --
DELETION -- WELL.
LASTLY, IF WE LEAVE THEM LONGER
AND THOSE MICE ARE SHOWING
REDUCED SURVIVAL, AND HAVE
SIGNIFICANT MORE FIBROSIS.
SO TO SUMMARIZE WHAT WE HAVE
RECENTLY WORKING ON DYSREGULATED
THE CELLULAR METABOLISM -- PLUS
PATHWAYS AS WELL AS -- AND WE
REALLY THINK THAT IT'S HARD TO
RECOVER TYPE 2 CELLS, POSSIBLE
TO RETORE TYPE 2 CELL PROGENITOR
CELL FUNCTION, WE PROBABLY NEED,
YOU KNOW, ZINC/SIRT1 AND
POSSIBLE OTHER REGULATORS --
OTHER COMPONENTS.
THANK YOU VERY MUCH.
LOOKING FORWARD TO THE
DISCUSSION.
>> THANK YOU SO MUCH.
SO JUST AS A RECAP, PLEASE POST
YOUR QUESTIONS IN THE CHAT AND
WE'LL ADDRESS THEM DURING THE
DISCUSSION PERIOD.
SO I'D LIKE TO INTRODUCE
DR. FRANK MCKEON FROM THE
UNIVERSITY OF HOUSTON, TE TEXAS
MEDICAL CENTER.
HE'S DIRECTOR OF THE SOMATIC
STEM CELL CENTER.
>> GOOD MORNING.
THANK YOU FOR INVITING ME.
I WILL BE TALKING ABOUT WORK
THAT I DO TOGETHER IN A SHARED
LAB WITH WA XIAN FOR THE ALMOST
LAST TWO DECADES NOW.
THE FOCUS OF THE LAB HAS BEEN
CLONING STEM CELLS.
WHAT WE'LL TALK ABOUT TODAY IS A
FRINGE AND SOME PEOPLE WILL
UNKINDLY CALL IT A LUNATIC
FRINGE AREA OF BIOLOGY OF THESE
PATHOGENIC STEM CELLS THAT ARE
SHOWING UP IN CHRONIC
INFLAMMATORY DISEASES IN THE
LUNG, BUT THEY'RE ALSO SHOWING
UP IN OTHER DISEASES AS WELL.
AND I THINK THIS IS PROBABLY A
MODEL FOR CHRONIC DISEASES IN
GENERAL.
I WILL TELL YOU THAT THESE CELLS
ARE PRE-EXISTING IN NORMAL LUNG.
AT LOW LEVELS, THEY'RE PRESENT
IN FETAL LUNG.
AND THEY'RE EPIGENETICALLY
FIXED.
SO IN THE NEXT SLIDE, IT'S JUST
MY DISCLOSURES.
AND THE NEXT SLIDE IS SOME WORK
THAT GOT US INTO THIS.
THIS IS WORK WE HAD SET UP A
MODEL OF OURS LARGELY BASED ON
THE WORK OF MARGARET HERAGE ON
RECOVERY OF PAISHEDZ FRR ARDS.
THAT TOGETHER WITH THE WORK WITH
PEDIATRIC NECROTIZING PNEUMONIA
SHOWED YOU COULD HAVE MASSIVE
LUNG DAMAGE AND YOU COULD
RECOVER FROM IT.
THE QUESTION IS WHAT WAS THE
MECHANISM OF THAT.
SO WE SET UP AN H1N1 INFLUENZA
MODEL IN SINGAPORE THAT WE COULD
DIAL IN THE AMOUNT OF DAMAGE.
SO WE COULD SHOW THAT THESE MICE
COULD RECOVER FROM 60% LUNG
DAMAGE TO A PRUS TEEN LUNG AT
THE END OF THE DAY.
I DON'T HAVE A POINTER, BUT WHAT
YOU SEE IS THE GREEN IS NORMAL
LUNG, AND YOU SEE THESE AREAS
THAT HAVE BEEN -- IN BLUE THAT
HAVE BEEN INFILTRATED, AND
LARGELY DENUDED OF ANY SORT OF
LUNG TISSUE REPLACED BY
NEUTROPHILS AND MACROPHAGES.
RIGHT IN THE 16 TESTIFY OF ALL
THAT WERE THESE KERATIN
5P63 CELLS, AND THEY ONLY SHOWED
UP IN THOSE REGIONS IN MASSIVE
AREAS.
IF YOU SEE THE BLOWUP OF THAT,
YOU CAN SEE THEY'RE FORMING
STRUCTURES THAT LOOK SIMILAR TO
ALVEOLI.
WE'VE ACTUALLY CHASED THOSE INTO
ALVEOLI, AND I'LL SHOW YOU THAT.
NOW IN BOSTON, WE WERE HEAVILY
INFLUENCED BY THE CLASSIC WORK
OF HOWARD GREEN.
HE WAS THE FIRST TO CLONE THE
EPIDERMAL STEM CELL THAT'S BEEN
HEAVILY USED IN REGENERATIVE
MEDICINE.
SO ON THE NEXT SLADE, YOU'LL SEE
WE ADAPTED THAT SAME TECHNOLOGY,
AND WE WERE ABLE TO CLONE THESE
P63 CELLS FROM ALL REGIONS OF
THE AIRWAY: NASAL, TRACHEAL AND
LUNG.
AND EACH ONE OF THOSE DIFFERED.
DIFFERED
SOMEWHAT, THEY HAD THEIR OWN
SIGNATURES, BUT THEY'RE LARGELY
P63 KERATIN 5 POSITIVE CELLS.
IN THE NEXT SLIDE, YOU CAN SEE
THAT -- YOU CAN DIFFERENTIATE
THEM, UPPER AIRWAY FORMS UPPER
AIRWAY EPITHELIUM UPON DIFFICULT
RENNIDIFFERENTIATION IN A 3D CU.
THE LUNG ONES GIVE RISE TO
DIFFERENT STRUCTURES.
THEY'RE ENRICHED IN TYPE 1 AND
TYPE 2 PNEUMOCYTE MARKERS.
THESE CELLS TURNED OUT TO BE
EXTREMELY VI VERSATILE.
ON THE NEXT SLIDE, YOU CAN SEE
WE CAN CLONE THESE, WE CAN PUT
THEM INTO -- BY SIMPLE
INTRATRACHEAL DELIVERY PUT THEM
DOWN THE TRACHEA.
IN THE NEXT SLIDE, YOU CAN SEE
THAT THEY, IN FACT, INCORPORATE
IT INTO H1N1 DAMAGED LUNG, AND
THEY DIDN'T INCORPORATE INTO ONE
THAT YOU DIDN'T DAMAGE.
SO THIS GAVE US A TREMENDOUS
SAFETY -- A SENSE OF SAFETY WE
COULD I THINK RATE.
INCORPORATE.IF YOU ZERO IN TO TY
FORMED LUNG, YOU CAN SEE THAT
THESE -- THE LACZ IS, IN FACT,
LABELING THOSE, THE ALVEOLI.
SO IT GAVE US THE OVERALL
IMPRESSION THAT P63 CELLS FROM
THE DISTAL AIRWAYS GIVE RISE TO
BONA FIDE ALVEOLI.
SO AT THIS POINT WE WERE GOING
WELL, MAYBE WE HAVE A CURE NOT
ONLY FOR ARDS BUT WE HAVE A CURE
FOR CHRONIC LUNG DISEASE.
BUT AT THE SAME TIME, WE WERE
HEAVILY INFLUENCED BY THE WORK
FROM CHILOSI AND WITSETT,
SCHWARTZ AND REYNOLDS, ALL OF
THEM SHOWING THAT THERE'S
P63 CELLS IN THESE LESIONS THAT
WERE SHOWING UP IN THESE CHRONIC
LUNG DISEASE.
JAY SHOWED US SOME VERY NICE
PICTURES YESTERDAY OF THESE
METAPLASIAS.
THOSE METAPLASIAS ARE ALL
SUBTENDED BY P63 POSITIVE CELLS.
SO ANYWAYS, WE DIDN'T THINK WE
COULD JUST SORT OF THROW NORMAL
LONG STEM CELLS DOWN INTO A
HIGHLY REMODELED COPD LUNG.
SO WE STARTED TO LOOK AT WHAT
WAS GOING ON IN THE COPD LUNG.
ON THE NEXT SLIDE, YOU CAN
SEE -- WE COULD READILY USE THE
SAME TECHNOLOGY, GENERATE
LIBRARIES FROM CONTROL PATIENTS,
CONTROL LUNGS, THESE ARE ALL
OBTAINED FROM THE TRANSPLANT
CENTERS.
AND COPD LUNGS.
THESE LIBRARIES ARE CLONES OF
P36 POSITIVE CELLS.
IF YOU DO THE SINGLE CELL
ANALYSIS, LOOK AT THE PIE CHART
AT THE BOTTOM LEFT.
IT DOMINATEIT'S DOMINATED BY A M
CELL IN BLUE.
THAT'S THE NORMAL STEM CELL THAT
GAVE RISE TO THIS REGENERATIVE
STEP.
IF YOU LOOK AT THE COPD, AND IT
DIDN'T MATTER WHICH PATIENTS FOR
COPD, WHICH TYPES, THEY ALL
LOOKED ALIKE, THEY WERE
DOMINATED BY THESE THREE WHICH
AT THE TIME WE CALLED VARIANTS.
AND IF YOU LOOK VERY CLOSELY ON
THE PIE CHARTS ON THE LEFT-HAND
SIDE, YOU SEE THAT EVEN THE
NORMAL PATIENTS HAVE SOME OF
THOSE CLONES THAT COME TO
DOMINATE THE COPD PATIENTS.
AND I SHOW YOU THIS SLIDE
BECAUSE THIS IS A PARADIGM FOR
EVERYTHING ELSE WE'VE SEEN, NOT
ONLY IN LUNG BUT ALSO IN
PEDIATRIC CROHN'S DISEASE AND
ULCERATIVE COLITIS.
YOU'RE GETTING THE SAME
EMERGENCE OF THESE VARIANTS.
SO THE QUESTION IS WHAT ARE
THESE VARIANTS DOING?
ON THE NEXT SLIDE, YOU CAN SEE
JUST A SIMPLE SHOTGUNNING OF
THOSE LIBRARIES INTO HIGHLY
IMMUNODEFICIENT MICE.
SO THESE MICE, THE ONLY THING
THEY HAVE IS NEUTROPHILS.
AND IF YOU LOOK AT THE CONTROL
ONES, THEY SELF-ASSEMBLE INTO
THESE LUMEN THAT ARE HOLLOW,
ESSENTIALLY CLEAR, WHEREAS THE
COPD, THEY'RE ALL FILLED WITH
NEUTROPHILS, AND THIS IS TRUE
ALL ACROSS THESE DIFFERENT
PATIENTS THAT WE LOOKED AT.
SO IT LOOKED AS IF THERE'S SOME
INFLAMMATORY COMPONENT OF THIS.
IF YOU LOOK IN THE NEXT SLIDE,
YOU GET MYOFIBROBLASTS
SURROUNDING THESE SAME LIBRARY
NODULES.
AND AGAIN, IT'S TRUE ACROSS ALL
THE COPD PATIENTS, IT'S NOT THE
CASE WITH CONTROL PATIENTS.
SO IN THE NEXT SLIDE, WE COULD
JUST USE SINGLE CELL SORTING TO
BREAK THESE LIBRARIES DOWN, AND
THEN CAN YOU FIND YOU COULD
MAKE -- CLUSTER ONE WAS THE
NORMAL STEM CELL.
CLUSTER TWO WAS A GOBLET CELL
METAPLASIA.
AND CLUSTERS THREE AND FOUR WERE
BOTH SQUAMOUS CELL METAPLASIA,
ONE OF WHICH IS FIBROTIC AND ONE
OF WHICH IS INFLAMMATORY.
AND THE GENE EXPRESSION PROFILES
EXACTLY MATCHED THESE PROFILES
HERE.
ON THE NEXT SLIDE, YOU CAN SEE
THAT WE'VE CARRIED THIS ON TO
IPF, AND TO CYSTIC FIBROSIS.
CYSTIC FIBROSIS IS ACTUALLY MORE
COMPLICATED.
IT'S GOT FIVE DIFFERENT
VARIANTS, THREE OF WHICH OVERLAP
WITH THOSE OF COPD.
FOR IPF, WE'RE STILL WORKING ON
THE HONEYCOMB -- WITH DAVID SH
WASCHWARTZ AND HIS COLLEAGUES BT
WE'VE PUBLISHED ON THE ONE THAT
CAN GIVE RISE TO CUBOIDAL
SYNDROME.
A THAT SAME TECHNOLOGY WE'VE
USED FOR SEVERE ASTHMA, FOR RA
AND COVID INTERSTITIAL LUNG
DISEASE.
WE DON'T KNOW HOW MANY OF THESE
VARIANT STEM CELLS EXIST BUT
WE'RE READY SURE AT THIS POINT
THAT THESE CELLS ARE PART OF
YOUR INNATE IMMUNE SYSTEM,
INNATE MUCOSAL IMMUNITY, AND
THEY HAVEN'T BENZENE BECAUSE BEE
THEY'RE PRESENT AT SUCH LOW
LEVELS.
BUT NEVERTHELESS, FROM OUR SIM
POLICE TI VIEW OF THE WORLD,
IT'S LIKE YOU HAVE GOOD GUYS AND
BAD GUYS, THE NORMAL LUNG STEM
CELL AND THEN YOU'VE GOT THESE
VARIANTS THAT ARE DRIVING THINGS
LIKE MUCOUS, HYPER NECROSIS AND
INFLAMMATION.
SO IT BECOMES A SIMPLE THING,
ALL OF THESE CELLS ARE
INTRINSICALLY IMMORTAL, SO YOU
CAN GROW THEM UP AT HIGH LEVELS
FOR HIGH THROUGHPUT SCREENS SUCH
AS 384 WELL SCREEN, SO ON THE
NEXT SLIDE, YOU CAN SEE WHAT
THOSE ARE LOOKING LIKE.
SO WE HAVE GENERATED SOME VERY
NICE AND VERY SPECIFIC, WORKING
TOGETHER WITH OUR CHEMISTRY
COLLEAGUES AT TUFTS, SOME
INCREDIBLY POWERFUL MOLECULES
THAT ARE PART OF A SYNTHETIC
LETHAL COMBINATION.
THE GOOD NEWS IS, THESE ARE
SHOWING BROAD EFFECTS ACROSS ALL
THESE VARIANT CELLS, AND WE CAN
GET INTO WHAT MAKES THESE
VARIANT CELLS EXPAND, BECAUSE I
THINK THAT'S THE KEY TO
UNDERSTANDING THEIR
VULNERABILITIES.
AND ON THE LAST SLIDE, I'LL JUST
SHOW YOU SOME OF THE KEY POINT
THAT I THINK ARE OUT THERE.
SO THERE'S CLEARLY A DOMINATION
BY THESE PATHOGENIC STEM CELLS
ACROSS ALL THESE CHRONIC LUNG
DISEASES, AND THAT EXTENDS INTO
CROHN'S AND UC, AND IT WOULDN'T
SHOCK ME THAT IT'S EXTENDING
INTO NASH AND CHRONIC KIDNEY
DISEASE.
THESE PATHOGENIC STEM CELLS
EXIST AT LOW LEVELS IN NORMAL
AND FETAL LUNG.
WE POSIT THEY REPRESENT
COMPONENTS OF THE INNATE
IMMUNITY.
WE'VE GOT TO DEMONSTRATE THIS.
AND THEN THE QUESTION IS, WHAT
CHANGES IN THE PATHOGENIC STEM
CELLS -- SO WE NORMALLY EXPECT
THESE THINGS TO SPIKE UP IN
RESPONSE TO A PATHOGEN AND THEN
DISAPPEAR, AND THEY DON'T.
WHAT WE CAN TELL FROM THE GENE
EXPRESSION PROFILES IS THERE'S
TWO THINGS THAT ARE GOING ON,
AND SOME OF THESE, DR. HAN
REFERRED TO EARLIER, THERE'S
SOMETHING TO DO WITH COMPLEX
1 IN THE MITOCHONDRIA AND THEN
THERE'S SOMETHING TO DO WITH
RESISTANCE TO FAST LIGAND --
CELL DEATH.
WE ALSO HAVE TO TRY TO CONNECT
UP THESE DATA WITH WHAT'S GOING
ON IN THESE REALLY BEAUTIFUL
SINGLE CELL RNA SEQ STUDIES OF
IPF, THESE SO CALLED ABNORMAL
BASALOID CELLS, AND I THINK
OTHERS HAVE MENTIONED THIS
ALREADY, THE VAST MAJORITY OF --
EVEN THOUGH 10 TO 20% OF HEAVY
SMOKERS DEVELOP COPD, THE VAST
MAJORITY OF CANCERS COME FROM
PATIENTS WITH PRE-EXISTING COPD.
SO ARE THESE PATHOGENIC STEM
CELLS SOMEHOW IN THAT TRACK TO
CANCER?
AND FINALLY, THIS DRUG
DEVELOPMENT.
WE REALLY NEED -- WE'VE GOT
100 MILLION PEOPLE JUST IN THIS
COUNTRY ALONE WITH CHRONIC LUNG
DISEASE.
WE REALLY NEED THERAPEUTICS.
WE'RE VERY EXCITED ABOUT WHERE
WE ARE NOW AND THERE'S A LOT OF
IND-ENABLING STUDIES GOING ON
NOW.
GOD WILLING, WEAR GOING TO GET
SOMETHING LIKE THIS INTO THE
NEXT -- AND I WANT TO THANK THE
PEOPLE ON THE NEXT SLIDE, SO A
WHOLE BUNCH OF GREAT
COLLABORATORS AND TALENTED
PEOPLE IN THE LABORATORY, AND
GREAT SUPPORT FROM THESE VARIOUS
PLACES.
THANKS VERY MUCH.
>> THANK YOU SO MUCH, FRANK.
I THINK WE CAN NOW OPEN THE
DISCUSSION.
AND START WITH MINGXIA.
CAN YOU TURN ON YOUR CAMERA?
>> THE QUESTION IN THE CHAT.
>> I WAS BASICALLY TRYING TO
UNDERSTAND MORE ABOUT THE
FUNCTIONAL TESTING OF THE IPF --
WE'VE BEEN STRUGGLING A LITTLE
BIT ON SHOWING THE RELEVANT
FUNCTION OF THE -- AND OTHER
CELL TYPES AND LUNG ORGANOIDS
AND HOW PRECISELY THOSE
FUNCTIONS ARE RELATED TO THE
PHENOTYPES THAT WE'RE SEEING IN
PATIENTS.
SO THAT WAS MY QUESTION TO
DR. WILSON.
>> SO I READ IT IN THE CHAT BUT
I CAN MAYBE JUST TRY TO EXPLAIN
VERBALLY AS WELL.
SO TRYING TO IDENTIFY FUNCTIONAL
BEHAVIORS OF PARTICULAR CELL
TYPES IN VITRO AND TRY TO RELATE
THEM TO IN VIVO BIOLOGY IS A
MAJOR CHALLENGE AND THAT WAS
SOMETHING WE HAD TO THINK A LOT
ABOUT WHEN WE WERE TRYING TO
UNDERSTAND THE CONSEQUENCES OF
PERTURBING INDIVIDUAL GENES IN
OUR SYSTEM, YOU KNOW, WE'RE
DEALING WITH A SINGLE IPSC
DERIVED CELL TYPE, IN THIS CASE
IA T2s, SO THE THINGS THAT WE
WERE LOOKING AT THAT WE THOUGHT
HAD SOME KIND OF FUNCTIONAL
RELEVANCE TO A PROGENITOR CELL
TYPE, AS I MENTIONED IN MY TALK,
WERE THINGS LIKE PROLIFERATION,
DIFFERENTIATION EFFICIENCY, AND
MATURATION.
WAND I SAID IN THE CHAT AND WHAT
WE DIDN'T LOOK AT AT THE TIME
WAS DIFFERENTIATION EFFICIENCY
IN TERMS OF GENERATING TYPE
1 CELLS, WOULD BE OBVIOUS
RELEVANT PHENOTYPE FOR A TYPE
2 CELL.
THE REASON WE DIDN'T DO SO AT
THE TIME WAS DARRELL'S LAB WAS
WORKING ON THAT TIME TO GENERATE
PROTOCOLS TO GENERATE IET1s
FROM IE T2s BUT THEY WEREN'T
REALLY PERFECTED YET.
BUT AT THIS POINT WE COULD DO
THAT NOW.
SO I THINK THAT WOULD BE ANOTHER
RELEVANT PHENOTYPE FOR A
PROGENITOR CELL LIKE A TYPE
2 CELL.
>> I GUESS I ASKED THAT QUESTION
MOSTLY BECAUSE THESE FUNCTIONS
ARE MORE GENERAL STEM CELL
FUNCTIONS, SO I WAS WONDERING IF
THERE'S ANY SPECIFIC FUNCTION,
ESPECIALLY THE FUNCTIONAL
DEFECTS THAT WE WOULD SEE IN A
COPD LUNG.
BUT I GUESS THAT'S THE --
>> SO I THINK THAT'S THE RIGHT
KIND OF QUESTION TO BE THINKING
ABOUT, AND OUR HYPOTHESIS WOULD
BE THAT THE PROGENITOR FUNCTION
WOULD BE SOMETHING THAT WOULD BE
IMPAIRED IN COPD OR EMPHYSEMA
BUT FOR SURE ALSO THINKING ABOUT
THE CROSSTALK BETWEEN DIFFERENT
CELL TYPES IS SOMETHING TO
CONSIDER AND WE HAVE THIS
REDUCTIONIST PLATFORM WHERE
WE'RE LOOKING AT ONE CELL TYPE.
YOU KNOW, PLATFORMS LIKE YOURS
THAT INCLUDE MULTIPLE CELL TYPES
MIGHT BE REALLY INTERESTING TO
TRY TO ASK THAT KIND OF
QUESTION, AND OUR PLAN IS TO
SORT OF BUILD IN COMPLEXITY OVER
TIME AS WE'RE ABLE TO BUILD
COCULTURAL SYSTEMS.
>> THANK YOU.
>> SORRY.
THERE ARE SOME HANDS RAISED IN
THE CHAT TOO.
SO CAN WE START WITH JIANWEN.
>> I HAVE QUESTION FOR DIANHUA,
MIXTURE OF MANY CELL TYPES, HAVE
YOU THINK ABOUT TO ISOLATE
PARTICULAR CELL TYPES INSTITUTE
INTO THE TRACHEA OR SYSTEMATIC
DELIVERY AND SEE WHETHER CERTAIN
TYPE OF MESENCHYMAL STEM CELLS
WOULD BE MORE CURATIVE, IF YOU
HAVE STRONG EVIDENCE TO SHOW
THAT.
>> SO THAT'S AN EXCELLENT
QUESTION.
AND LET ME EXPAND ON THAT
BECAUSE THE MSCs THAT ARE
ISOLATED FROM BONE MARROW VERSUS
PLACENTA VERSUS ADIPOSE ARE
SIMILAR BUT NOT COMPLETELY
IDENTICAL.
AND FURTHER, WHEN YOU, FOR
EXAMPLE, GET AN EXTRACTION,
LET'S SAY, A BONE MARROW ISOLATE
LIKE THAT IN A CULTURE SYSTEM,
THEN THERE CAN BE CLONES THAT
DEVELOP.
OF MSCs THAT WILL DEVELOP
DIFFERENT ATTRIBUTES, DIFFERENT
PROPERTIES AND THE LIKE TOO, SO
IT'S A DIFFICULT QUESTION, AND
THERE HAVE BEEN A NUMBER OF
COMPARATIVE STUDIES IN
PRE-CLINICAL MODELS, NOT JUST
LUNG DISEASE BUT A RANGE OF
OTHER INDICATIONS COMPARING, FOR
EXAMPLE, ADIPOSE VERSUS PLACENTA
ORIGIN.
AND THERE ARE SOME DIFFERENCES
IN THE OUTCOME, BUT THAT HASN'T
YET TRANSLATED INTO A RATIONAL
APPROACH TO CLINICAL
INVESTIGATION.
WHEN YOU LOOK AT IT
HISTORICALLY, THE MAJORITY OF
TRIALS HAVE UTILIZED BONE MARROW
ISOLATED MSCs CULTURED UNDER
NORMOX IC CONDITIONS WITH
CERTAIN GROWTH FACTORS IN THE
MEDIA AND THE LIKE TOO.
PART OF THAT IS PRACTICALITY,
PART OF THAT IS ACCESSIBILITY.
BUT WHEN YOU LOOK, FOR EXAMPLE,
AT THE LARGE NUMBER OF TRIALS
THAT STARTED IN THE PANDEMIC FOR
COVID-ASSOCIATED ARDS, THE
MAJORITY OF THOSE UTILIZE THE
UMBILICAL CORD ORIGIN MSCs,
AND THE REASON THERE IS THAT
THOSE ARE EASIER TO OBTAIN THAN
DOING AN ADIPOSE LIPOSUCTION OR
A BONE MARROW ASPIRATE.
AND SO IT'S THE SCIENCE VERSUS
THE PRACTICALITY VERSUS IF
YOU'REYOU'RE AN INDUSTRY PARTNE,
YOU'RE GOING TO WANT TO GO FOR
THE MOST DIRECT GROUP.
SO THERE'S STILL A LOT OF ROOM
TO IMPROVE UPON THAT, AND THERE
IS NO RIGHT -- THIS IS
IMPORTANT, THERE'S NO RIGHT,
THERE'S NO WRONG, BUT AT THE END
OF THE DAY, I STILL THINK THAT
THE INFLAMMATORY PHENOTYPE IS
REALLY GOING TO BE THE MAJOR
DETERMINING FACTOR AND WILL
OVERSHADOW, IF YOU WILL,
INDIVIDUAL DIFFERENCES BETWEEN
DIFFERENT ISOLATES OF CELLS.
>> THANK YOU.
>> WHILE WE'RE GOING THROUGH THE
CHAT, COULD I ASK A QUESTION TO
DIANHUA?
>> YES.
>> I HAVE TWO QUESTIONS, REALLY.
THE FIRST IS, YOU SAID ALL THIS
GREAT DATA ABOUT THE COLONY
FORMING EFFICIENCY AND GROWTH
POTENTIAL OF CELLS AS A FUNCTION
OF THE INTRACELLULAR ZINC, AND
I'M WONDERING WHETHER, SINCE IN
YOUR OWN EARLY STUDIES SHOWED
THE SORT OF LOSS OF INTEGRITY OF
IPF TYPE 2 CELLS, WHETHER THIS
GROWTH IS ACCOMPANIED BY CHANGES
IN DIFFERENTIATION OR THERE'S A
REVERSION TO A NORMAL TIP 2 CELL
OR IS IT A HETEROGENEOUS
POPULATION IN IPF AS IT IS TO
BEGIN WITH?
>> THAT'S A REALLY GOOD
QUESTION.
WE DO NOT KNOW.
I MEAN, YOU KNOW, SO WHAT I'M
GUESSING IS THAT -- IPF FOR
LUNGS YOU HAVE REALLY 10%, 15%
CELLS ARE REALLY -- POSITIVE --
SO WE CALL THEM -- WE DO NOT
KNOW IF -- IF THIS IS ZINC,
INCREASES POPULATION, VERY
LIKELY, AND WE HAVE ANOTHER
THING I THINK IS PROBABLY MORE
IMPORTANT IS THAT EVEN SOME
CELLS THAT LOOK NOT EXPRESS A
HIGH -- MAY RECOVER SOME OF
THEIR FUNCTIONALITY IN TERMS OF
3D -- MATRIGEL SYSTEM.
I MEAN, IT'S REALLY HARD TO
CHARACTERIZE THAT, AND WE
HAVE -- DON'T HAVE A GOOD MEANS
TO REALLY EXPLORE THAT.
I'M NOT SURE IF I ANSWERED YOUR
QUESTION.
>> YEAH, MAYBE SEND ME SOME
ORGANOID OR PCS SYSTEM THAT
WOULD PROVIDE A CULTURE SYSTEM
WHERE YOU COULD SEE A CHANGE IN
TRANSCRIPTIONAL STATE MIGHT
HELP.
MY QUESTION IS, ZINC IS CHEAP.
ARE YOU PLANNING TO TAKE THIS TO
A CLINIC AND IF YOU'RE TRYING TO
DO THAT, COULD YOU HIGHLIGHT
WHERE THAT STAND?
>> RIGHT, THAT'S A SIGNIFICANT
INTERESTING -- AFTER I PUBLISH
THE PAPER AND IT WAS A LOT OF --
CAN WE USE ZINC SUPPLEMENTS FROM
AMAZON OR SOMEWHERE -- CVS AND
WE CAN USE THEM FOR THE -- RIGHT
NOW OURSELF WE DO NOT HAVE ANY
PLANS TO DEVELOP.
IF WE LOOK AT A PAPER -- WHAT WE
REALLY THINK, YOU FEED SOME SORT
OF COMBINATION THERAPY, HOWEVER,
SOME TYPES OF FUNCTIONALITY, YOU
CAN USE A PROPER COMBINATION
WITH ZINC, WITH SOME OF THE
ACTUATORS OR -- SIRT1 -- A LOT
AVAILABLE FOR THAT -- SO -- IT
DEFINITELY WORTH IN CLINICAL
TRIAL TO LOOK INTO -- IF YOU CAN
RECOVER SOME OF THE -- FUNCTION
NOT JUST SOME OTHER --
>> SHOULD WE START WITH
WELLINGTON?
HIS HAND HAS BEEN UP FOR A
WHILE.
HIS ARM MUST BE VERY TIRED.
ELK TON?
WELLINGTON?
>> HE FELL ASLEEP WAITING.
>> XIAN --
>> WHATEVER YOU SHOW IS QUITE
SURPRISING AND I WAS WONDERING,
WHEN YOU TALK ABOUT THESE
CLONES, AND YOU MENTIONED -- I
REMEMBER YOU HAVING -- YOU WERE
MENTIONING THAT THERE IS THE
CELL SURFACE MARKERS THAT YOU
ARE LOOKING FOR IN ORDER TO GET
SOME OF THIS EXTRACTED INTO THE
MADE ORGANOIDS AND EXPLORE
FURTHER.
SO DO YOU HAVE CELL SURFACE
MARKERS FOR AT LEAST SOME OF
THOSE THAT ONE CAN SEPARATE?
>> WE'VE UPLOADED THEM INTO
THE --
>> YES, I SEE.
I SEE.
AND WHEN YOU TALK ABOUT THE
CLONE -- THESE ARE SOMEHOW CELL
AUTONOMOUS, HOW DO YOU THINK
THAT THIS WORKS, PARTICULARLY
WHEN WE THINK THAT THESE ARE
EARLY PRECURSORS OF THE COPD
LESIONS THAT AS ANDREW SAYS,
IT'S BETTER THAT WE FOCUS ON THE
EARLY STAGES THAN THE LATE.
>> RIGHT, RIGHT, RIGHT.
WELL, YOU KNOW, ONE THING WE
HAVEN'T QUITE DONE IT FOR THAT,
BUT -- LOOKED AT ADAPTING THE
SAME CLONING TECHNOLOGY FOR BOTH
BOWEL AND PLEURAL EFFUSIONS TO
LOOK ACTUALLY AT THE CANCER
REALM, AND IT TURNS OUT,
COMPARED TO WHAT THEY USUALLY
DO, WHICH IS GET THESE THINGS
CENTRIFUGED TO THE CELLS AND TRY
TO GET A PATHOLOGICAL READOUT ON
THEM, SIMPLY CLONING THE CELLS
WAS A MUCH BETTER APPROACH TO
IT.
SO IT MAY BE THAT WE CAN TAKE --
AND USE THESE TYPES OF CELLS AS
A READOUT PERHAPS FOR EARLY
DISEASE.
AND I THINK WE SHOULD PROBABLY
PUT OUR HEADS TOGETHER ON THIS
AND FIGURE OUT HOW TO DO THIS
SYSTEMATICALLY.
BUT IT WORKED FOR THE CANCER.
THERE'S A WHOLE THING ON LUNG
CANCER THAT'S GOING ON, BUT WE
SHOULD -- SOMEHOW THIS GROUP
SHOULD FIGURE OUT HOW TO GET
THIS DONE IN A SYSTEMATIC WAY TO
SAY WHAT IS THE FREAK SEE OF
NFREQUENCY OFNORMAL CLONES VERSC
CLONES DERIVED FROM -- AS YOU
KNOW YOU'RE EXPLORING A LOT OF
REGIONS OF THE LUNG WITHOUT
HAVING TO -- WELL, I GUESS --
>> THANK YOU.
>> COULD I HAVE A FOLLOW-UP
QUESTION ALONG THOSE LINES?
SO I'M TRAYING TO UNDERSTAND,
WHAT IS -- WHAT STEAMS UNIQUE
ABOUT THESE CELLS, AT LEAST ONE
THING, IS THAT YOU THINK THEY'RE
EPIGENETICALLY FIXED.
>> YES.
>> AND THAT'S A LITTLE UNUSUAL
FOR STEM CELLS TO BEGIN WITH,
BUT I GUESS I'M WONDERING
WHETHER THAT SUCCESSION IMPLIES
SOME -- IT REALLY REFLECTS A
MEMORY FROM SOME FORMER STATE
THAT NOW --
>> ONE THING THAT WA IS ABOUT TO
PUBLISH ON IS THE EPIGENETIC
PROFILES OF THE ENTIRE GI TRACT,
FROM STOMACH TO RECTUM, AND EACH
OF THOSE, CLONOGS ENIC AND
ABSOLUTELY FIXED -- I HAD THE
FEELING THAT THESE CELLS -- BUT
WE HAVEN'T DONE THE EXPERIMENTS
TO SHOW THIS.
BUT I HAVE THE FEELING THAT'S
CELLS ARE PART OF THIS INNATE
IMMUNE SYSTEM.
THEY COORDINATE INFLAMMATION,
THEY COORDINATE FIBROSIS TO WALL
OFF PATHOGENS, THEY GUM THEM UP
WITH -- I THINK THAT'S THEIR
NORMAL ROLE.
AND I THINK THEY'VE BEEN FIXED
TO BE THAT WAY, BUT WE'RE SEEING
THEM IN EVERY ORGAN THAT WE LOOK
AT.
FOR INSTANCE, FOR TERMINAL ILEUM
AND PEAD TR PEDIATRIC CROHN'S.
THEY'RE THERE AHEAD OF TIME
BUT --
>> DO YOU HAVE A WAY TO IDENTIFY
THOSE IN SPACE, IN SITU, WITHOUT
PULLING THEM OUT AND SUBCLONING
THEM?
>> HERE'S THE THING, IT'S
COMPLICATED.
THESE CELLS REPRESENT ONE OUT OF
A THOUSAND OF ALL EPITHELIAL
CELLS ARE CLONOGENIC.
THAT NUMBER IS PRETTY FIXED
ACROSS TISSUES, WHETHER IT'S GI
TRACT, LUNG OR ANYTHING ELSE.
AND THEN, OF COURSE, WHEN YOU DO
A SINGLE CELL RNA SEQ FOR THE
WHOLE LUNG, IT MOSTLY IMMUNE AND
OTHER CELLS, SO YOU'RE LOOKING
AT A MAYBE ONE OUT OF 10,000
CELLS, IS THIS BASALOID
COMPONENT.
WHEREAS WE'RE AMPLIFYING THAT
REGION TO A GREAT -- THAT'S ALL
WE'RE CLONING.
SO I THINK THAT'S THE -- IT SORT
OF A -- MYOPIC IN THE SENSE YOU
DON'T HAVE ANY OTHER INFORMATION
THERE, BUT YOU DO HAVE THOSE
CLONOGENIC CELLS AND YOU'RE
AMPLIFYING THEM.
THEY BEHAVE VERY WELL.
I MEAN, THEY'RE IMMORTAL.
YOU CAN JUST GROW THEM FOREVER
AND AT THE END OF -- WE'VE DONE
SOME WE'VE GROWN CONTINUOUSLY IN
THE LAB FOR A YEAR, AND WHEN YOU
ASK THEM TO DO SOMETHING, THEY
ALWAYS DO THE RIGHT THING.
THEY DON'T FORGET.
WA IS DOING A LOT OF -- SHE'S
DONE ALL THE EPIGENETICS ON THE
GI TRACT BUT SHE'S NOW DOING
THAT WITH THE LUNG.
SO WE'LL BE ABLE TO SORT OF
FURTHER NAIL DOWN --
>> IT STILL SEEMS LIKE IT'S A
CHALLENGE TO FIGURE OUT WHERE
THEY -- EXACTLY WHERE THEY ARE
IN SITU BECAUSE OF THE RARITY.
IS THAT FAIR?
>> THAT'S FAIR, BECAUSE THEY'RE
RARE CELLS OF RARE CELLS.
BUT FROM FETAL TISSUE, FOR
INSTANCE, THEY'RE A LITTLE BIT
MORE COMMON THAN THEY ARE
FROM -- IN NORMAL LUNG.
IN FETAL TISSUE, THEY'RE
ACTUALLY, YOU KNOW, MAYBE
UPWARDS OF 10% AT 22 WEEKS.
>> IT WILL BE REALLY INTERESTING
TO FIGURE OUT HOW IT IS THAT
THEY'RE EPIGENETICALLY FIXED.
>> YES, YES.
WELL, I KNOW SHE'S GOTTEN --
GOING -- ON THAT RIGHT NOW, IT
WOULD BE REALLY FUN TO SEE, I
AGREE WITH YOU.
>> SO I WANTED TO ASK AN DRAW,
THEORETICALLY BASED ON SOME OF
THE CLINICAL TRIALS THAT DAN HAS
TALKED ABOUT AND SOME OF THE
BACKGROUND INFORMATION FROM
THAT, HOW WOULD YOU SEE
INTEGRATING IPSC AS A
THERAPEUTIC GIVEN HOW THE
CLINICAL TRIALS HAVE GONE WITH
CELL THERAPIES?
>> WELL, I GUESS I CAN IMAGINE
SOME POSSIBILITIES.
SO FIRST OF ALL, I GUESS I
SHOULD JUST MENTION SPECIFICALLY
THAT IN TERMS OF TRYING TO
ENVISION IPSC CELLS AS A
THERAPY, NOT IN THE CONTEXT OF
WHAT DAN TALKED ABOUT, BUT I
THINK WHAT DARRELL'S GROUP HAS
SHOWN IN HIS RECENT PUBLICATIONS
IS THE POTENTIAL FOR IPSC
DERIVED EPITHELIAL CELLS TO
ENGRAFT EITHER IN THE TRACHEA OR
THE DISTAL LUNG SO THAT'S BEING
PERFORMED SPECIFICALLY WITH AN
EYE TOWARDS A FUTURE CELL-BASED
THERAPY FOR EPITHELIAL CELLS AND
I WOULD ENVISION HE WOULD TRY TO
APPLY SIMILAR TECHNIQUES TO
OTHER CELL TYPES AND I CERTAINLY
CAN IMAGINE IF ONE CAN
GENERATE -- I'M NOT SURE IF
THERE'S A NEED TO DO THAT SO
MUCH FOR MESENCHYMAL CELLS BUT I
CAN IMAGINE DEVELOPING
MESENCHYMAL CELLS SIMILAR TO THE
PROPERTIES THAT DAN HAS TALKED
ABOUT, ALTHOUGH WE CAN GET THOSE
PRIMARY CELLS RIGHT NOW, SO I
GUESS THE RATIONALE FOR THE IPSC
DERIVED VERSION MAYBE IS NOT
QUITE AS CLEAR TO ME.
BUT FOR EPITHELIAL CELLS, DAN,
YOU LOOK LIKE YOU HAVE SOMETHING
TO ADD TO THAT.
>> WELL, SO JUST TO GO OFF ON
ONE POINT, ANDREW, THERE IS A
PUSH TOWARDS UTILIZING IPSC
DERIVED MSCs FOR THERAPEUTICS.
FOR THE PREVIOUS QUESTION,
CONTINUE TO HIGHLIGHT SOME OF
THE DIFFICULTIES IN THE FIELD OF
HETEROGENEITY OF THE PRODUCTS
THAT CAN BE ISOLATED FROM THE
DIFFERENT SOURCES, AND SO IPSC
DERIVED MSC HAS ALL THE
ATTRIBUTES OF BEING A UNIFORM
POPULATION, DOUBLING TIMES,
EXPANSION POTENTIAL, LACK OF
TERATOGEN IS IT, ET CETERA, SO
IT'S A REAL POSSIBILITY.
BUT THE GOAL -- TWO PARALLEL
GOALS AS YOU ARTICULATE, ONE IS
STRUCTURAL REPAIR, ONE IS
FUNCTIONAL MODULATION FOR AN
APPROACH FOR AN INFLAMMATORY OR
AN AUTOIMMUNE CONDITION IN WHICH
STRUCTURAL REPAIR ISN'T THE
PRIMARY GOAL.
SO APPLES AND ORANGES TO SOME
DEGREE.
>> NICELY STATED.
>> JOHANNS.
JOHANNES.
>> THANK YOU FOR THE NICE
PRESENTATIONS.
MY QUESTIONS ARE FOR FRANK.
ALONG THOSE LINES, THAT WAS IN
DISCUSSION ALREADY.
SO WHEN THE CLONES ARE TAKEN,
SAY, FOR EXAMPLE, FROM THE SAME
LUNG, DIFFERENT REGIONS, DISEASE
REGION, NORMAL REGION OR LARGE
AIRWAY, SMALL AIRWAY, DO YOU SEE
THE CLUSTER DIFFERENCES WITHIN
THE SAME LUNG OR IS IT PRAIMLY
DRIVEN WITHIN ONE CLUSTER FOR
THOSE CLONES THAT HAVE COME FROM
THE LUNG?
>> WE HAVEN'T DONE THIS
RELIGIOUSLY.
AND YET WE'RE GETTING -- SOME
AREAS ARE EASY TO PICK OUT, THE
HONEYCOMB REGIONS ARE EASY TO
PICK OUT AND THE SCHWARTZ LAB
HAS DONE THAT.
AND SENT US THOSE TISSUES.
SO THOSE ARE THINGS THAT YOU CAN
RECOGNIZE.
IF WE GET THESE LUNGS AND WE'RE
SORT OF OVERWHELMED, WE'RE
TRYING TO FIND THE UPPER
REGIONS, FOR INSTANCE, IN A COPD
LUNG THAT ARE DAMAGED, AND --
BUT WE SHOULD BE DOING IT AS YOU
SAY IN A MORE SYSTEMATIC WAY.
WE JUST DON'T HAVE THE BANDWIDTH
FOR DOING IT, BUT IT WOULD BE
VERY INTERESTING TO SAY THIS
LOOKS NOMINALLY NORMAL, WHAT
DOES THE PROFILE LOOK LIKE.
AND CAN YOU SEE -- IT GETS NA
THESE OTHER QUESTIONS ABOUT
SPREADING THAT PEOPLE WERE
TALKING ABOUT IN YESTERDAY'S
SESSION.
>> THAT'S RIGHT.
ALONG THE LINES THAT WE HEARD
YESTERDAY AFTERNOON AS WELL,
EVEN THE LARGE AIRWAYS SEEM TO
BE KIND OF NOT LOOKING VERY GOOD
FOR PEOPLE PRE-COPD.
SO THERE SEEMS TO BE A MAJOR
ISSUE WITHIN THE ENTIRE LUNG
WHEN IT COMES TO THESE KIND OF
ISSUES.
>> YOU KNOW, YOU'RE ABSOLUTELY
RIGHT.
IN FACT, ALL THE STUFF WE'VE
DONE WITH ASTHMA HAS BEEN
LARGELY WITH BRUSHINGS AND
BRONCHOSCOPY.
SO WE'RE GETTING JUST THE LARGE
AIRWAY.
SO IT'S A WHOLE AREA THAT WE
REALLY HAVEN'T TRIED TO MATCH UP
WITH WHAT'S GOING ON IN THE
LUNG, PER SE.
>> GREAT.
THANK YOU.
>> GREAT, THANK YOU.
>> OKAY, WE STILL HAVE SOME
HANDS UP IN THE CHAT.
>
>> SEUNGHYE?
>> YOU PRESENTED NICE DATA TODAY
ABOUT THE ZINC METABOLISM
RELATED TO THE NAD DEPLETION,
AND THAT ACTUALLY SOMEWHAT WAS
KIND OF CONSISTENT WITH I FIND
YESTERDAY, THAT THE LACK OF ANY
REGENERATION TRIGGERS PATHOLOGIC
ISR TO CHANGE THE STEM CELL
FATE.
SO I'M WONDERING IF YOU HAVE
LOOKED AT THE ISR ACTIVATION IN
YOUR -- -- TRY THE -- INHIBITOR
IN YOUR MICE.
>> RIGHT.
WELL, SO OUR STUDY, WE -- IF WE
LOOK INTO -- -- CELLS AND WE DO
SEE SOME OF THE STRESS RESPON
RESPONSE, -- OXIDATIVE STRESS.
ESPECIALLY IF WE LOOK INTO --
POSITIVE AND NEGATIVE CELLS SO
DEFINITELY WE SEE -- REACTIVE
OXYGEN RELATED RESPONSE AND THE
MITOCHONDRIA DYSFUNCTION AND WE
HAVE NOT TRIED TO USE --
COMPOUND AND WE -- SO -- M --
NPT LOSS AND WHAT HAPPENED, SO
WHAT WE'RE THINKING IS THAT
WHERE POSSIBLE, YOUR SYSTEM
IS -- VERY ACUTE SYSTEM IF YOU
HAVE WIRING -- AND THAT VERY
POSSIBLE WILL -- ISR RESPONSE
THAT YOU HAVE -- SUDDENLY YOU
HAVE A SIGNIFICANT DEMAND OF ATP
AND -- IN THE -- OF IPF, WE ARE
THINKING THERE WOULD BE SOME
SORT OF STRESSOR, LIKE SMOKING
OR SOME ENVIRONMENTAL STRESSORS.
THAT ACCUMULATES PROBABLY A
VERY, VERY LONG TIME, LET'S SAY
SOMETIMES 15 YEARS AND YOU HAVE
SIGNIFICANT REDUCED FOR SOME OF
THE -- EXCHANGE WHILE -- PK1.
I THINK THAT STRESS RESPONSE IS
PROBABLY A REAL WORRY LONG TERM
AND THE LOSS OF THAT GENE
EXPRESSION AND EVENTUALLY YOU
HAVE LOSS OF TYPE 2 CELL
FUNCTIONALITY.
THAT'S -- SO -- SO DEFINITELY
THAT SHARES SIGNIFICANT
SIMILARITY BETWEEN ACUTE FIRING
FUNCTION AND -- IPF -- OVER KIND
OF 15 EVEN LONGER YEARS.
>> BECAUSE I WAS WONDERING, YOU
MENTIONED WITH THE -- NAMP
KNOCKOUT MICE YOU BASICALLY
INDUCED MITOCHONDRIAL
DYSFUNCTION AND DEFINITELY
PROBLEM INDUCED THE HIGH RATIO,
SO I WAS WOULD BE WONDERING IF
THE ISR IS A DOWNSTREAM PATHWAY,
BECAUSE IN OUR MODEL, WE
ACTUALLY ADMINISTERED NMN WHICH
IS A PRECURSOR OF NAD AND WE
WERE ABLE TO RESCUE THE TOTALITY
AND THE MUSCLE WAS -- SO THAT'S
JUST MY THOUGHT.
>> WE THINK THAT WHERE POSSIBLE
IS -- THEY ARE VERY, VERY
POSSIBLE -- OXIDATIVE STRESS,
OTHER VERY RESPONSE AND --
RELATED.
WE HAVEN'T TRIED YOUR SYSTEM
USING INHIBITORS --
>> MAYBE WE SHOULD GET TO THE
OTHER HANDS IN THE CHAT.
YOU'VE BEEN WAITING A WHILE.
IF NOT, JOSE.
>> I'LL JUMP IN QUICKLY.
SO I THINK REALLY FASCINATING
DISCUSSION HERE.
JUST TO KIND OF JUMP ON THIS
IDEA OF THE CLONAL POTENTIAL OF
SOME OF THE STEM CELLS AND HOW
FIXED SOME OF THE -- COMMITMENT
MIGHT BE.
I WAS VERY CURIOUS FROM YOUR
TALK, FRANK, YOU'VE MENTIONED A
FEW TIMES THAT THESE CELLS MIGHT
BE PART OF SOME KIND OF INNATE
IMMUNE SYSTEM AND AS AN
IMMUNOLOGIST, IT KIND OF PIQUED
MY INTEREST IN TERMS OF HOWL
YOU'RE THINKING ABOUT THIS,
AND -- THEY MAY HAVE SOME
BENEFIT DURING AN ACUTE VIRAL
INFECTION OR DURING SOME KIND OF
BACTERIAL INFECTION, BUT THEN IN
THESE CHRONIC DISEASES, THEY'RE
BECOMING KIND OF COOPTED AND
EXPANDING TOO FAR OR KIND OF HOW
ARE YOU THINKING ABOUT THE
IMMUNOLOGICAL ROLE AND THEN THE
FUTURE DOWNSTREAM EXPANSION AND
PATHOGENESIS?
>> I THINK THE BEST -- WHAT'S
DIFFERENT BETWEEN THOSE WE DRAW
FROM FETAL AND NORMAL LUNG
VERSUS THOSE THAT ARE DOMINANT
IN THESE DISEASE STATES.
AND I BRIEFLY MENTIONED IT BUT
THERE'S A STRONG INHIBITION OF
FAST LIGAND INDUCED CELL DEATH,
AND AS YOU KNOW FROM YOUR
IMMUNOLOGY, THAT SEEMS TO BE A
BIG PLAYER IN GETTING RID OF T
CELLS YOU NO LONGER NEED.
SO I THINK THERE'S A LOT OF
SIMILARITIES THERE.
AND THERE'S SOMETHING ELSE
THAT'S GOING ON.
I'M TRYING TO WRAP MY HEAD
AROUND MITOCHONDRIA, WHICH IS
NOT A TRIVIAL THING, AND A LOT
OF THE GREAT TALKS IN THIS
SESSION WERE IN THIS MEETING ON
MITOCHONDRIA, BUT THERE'S
SOMETHING GOING ON WITH THE MY
TMITOCHONDRIA AND I THINK THOSE
ARE SOMEHOW ALSO PROTECTING
THESE CELLS FROM WHAT THEY --
INSTEAD OF SPIKING, THEY'RE JUST
PLATEAUING.
AND I THINK THAT PROBABLY -- I
DON'T KNOW, BUT THE GOOD NEWS
IS, IT'S THE SAME ACROSS ALL OF
THEM, WHETHER YOU LOOK IN IPF OR
CF OR COPD, THEY ALL SEEM TO
BE -- THIS CELL DEATH IN A
SIMILAR MECHANISM.
SO WE'RE REALLY EXCITED ABOUT
THAT TOO.
>> I GUESS ANY IDEA FOR THEIR
EXPANSION OR SELECTION IN MORE
ACUTE FORMS OF LUNG INJURY?
I KNOW YOU KIND OF BEGAN THE
TALK WITH SOME OF THE CLASSIC
STUDIES WITH THE H1N1 DAMAGE
MODEL AND MICE.
I KNOW IT'S ALWAYS HARD TO
CORRECT THE MOUSE AND THE HUMAN,
BUT ANY OPPORTUNITY TO STUDY
THIS IN MORE ACUTE VIRAL
SETTINGS?
>> NO, BUT I'M HEARING SOME
COLLEAGUES FROM M.D. ANDERSON
ARE DOING SORT OF CHRONIC
CHALLENGES WITH BACTERIA NA
LUNG, AND WITH TIME, THEY CAN
GET A CHRONIC DISEASE OUT OF
THAT, SIMILAR TO THIS SEQUENCES
OFBLY OWE MY 16 THAT RADANTE AND
OTHERS ARE TALKING ABOUT WHERE
YOU CAN GET A CHRONIC AND
PROGRESSIVE IPF.
THOSE ARE VERY INTRIGUING.
WHATEVER THEY'VE DONE, WE WOULD
IMAGINE THEY'VE GONE OVER THAT
HURDLE TO GIVE YOU STABLE CELLS
THAT ARE DRIVING DISEASE.
>> THANK YOU.
>> GREAT.
THANK YOU.
>> SO TALKING ABOUT PROGENITORS
AS INNATE IMMUNE CELLS FROM A
VASCULAR PERSPECTIVE, WE SEE
SIMILAR CHANGES IN ENDOTHELIAL
CELLS IN THE CAB LEARS AND IN
RESPONSE TO INFLAMMATORY
SIGNALING, THEY ACTUALLY CHANGED
THEIR FATTY ACID BINDING
REPERTOIRE SO THINKING ABOUT
THAT WITH METABOLISM AND -- TO
INTEGRATE THE METABOLISM AND --
HAVE YOU LOOKED AT FATTY ACID
BINDING PROTEINS AS WELL?
>> WHO, ME?
>> OH, FRANK.
SORRY, YEAH.
>> WE ARE LOOKING AT WHAT OTHER
THINGS THESE DO BESIDES EITHER
PRODUCING LARGE AMOUNTS OF MUCIN
OR DRIVING MYOFIBROBLASTS OR THE
IMMUNE CELLS.
THERE IS A LOT OF SIGNALING TO
ENDOTHELIAL CELLS.
AND SO WE'RE ESPECIALLY
INTERESTED IN THAT, YOU KNOW,
FOR PULMONARY HYPERTENSION AND
SUCH THINGS.
SO THAT, WE HAVE -- THAT WOULD
BE A REALLY FUN THING TO THROW
INTO.
BUT I THINK SOME OF THIS
PULMONARY HYPERTENSION MAY BE
GOING ON IN ALL OF THESE
CONDITIONS, SO HOW THEY INTERACT
WITH ENDOTHELIAL CELLS WOULD BE
SOMETHING WE'RE VERY INTERESTED
IN.
>> -- HAD THEIR HAND UP FOR A
LONG, LONG TIME.
XINWANG?
>> I THINK THEY MIGHT HAVE
STEPPED AWAY.
I'VE BEEN TEXTING THEM IN THE
CHAT.
SO MAYBE TOWARD THE END OF THIS
SESSION AND DISCUSSION, WE COULD
DISCUSS ADDITIONAL AREAS FOR
INVESTIGATION AND CURRENT
CHALLENGES, FUTURE STUDIES.
>> CAN I ASK ONE MORE QUESTION?
>> OF COURSE.
SORRY.
[LAUGHTER]
SO THIS IS FOR DR. MCKEON.
I KNOW YOU MENTIONED
SPECIFICALLY THAT YOU SEE SOME
EVIDENCE OF THE MITOCHONDRIAL
COMPLEX 1 ON THIS THING.
CAN YOU JUST SHARE A LITTLE BIT
MORE ON THE WHY YOU THINK THE
COMPLEX 1?
BECAUSE THE OTHER THING IS THAT
WHAT I'M REALIZING IS THAT EACH
MITOCHONDRIAL ETC COMPLEXES
MIGHT HAVE DIFFERENT ROLE AND
DIFFERENT FUNCTION BECAUSE WHEN
I USE THE -- I DO SEE ALL
DIFFERENT PHENOTYPE.
SO I'M JUST WONDERING WHAT'S
YOUR -- TO SHARE A LOT OF IT
MORE ABOUT THAT.
>> I WAS ABOUT TO CALL YOU AFTER
THIS SESSION, BECAUSE -- SO I
MEAN, THE ONE GENE -- ONE OF THE
GENES THAT'S SIGNIFICANTLY
UPREGULATED, AND AMONGST THE TOP
FEW, ACROSS THESE PATHOGENIC
CELLS IS A GENE CALLED NDUFA4L2.
AND YOU KNOW, ALREADY ITS NAME
ALONE IS ENOUGH TO BE ANNOYING,
BUT -- AND IT'S PART OF THE HIF.
AND IT HAS EFFECTS, AS YOU KNOW,
MITOCHONDRIAL RESPIRATION.
I HATE MITOCHONDRIA AND I HAVE
TO LEARN TO LOVE THEM AND THE
NOTION THAT YOU'VE GOT TO -- YOU
HAVE ALL THESE BEAUTIFUL SLIDE
AND -- YOU KNOW.
IT WOULD BE EMBARRASSING TO TALK
TO YOU BUT I WOULD LIKE TO SPEND
A LITTLE BIT OF TIME TO TRY TO
FIGURE OUT WHAT THAT GENE IS
DOING.
BUT IT'S VERY UPREGULATED, AND
IT'S -- OUR SUSPICION IS IT'S
PLAYING SOME ROLE IN THERE.
WE DON'T KNOW IF IT'S SHUTTING
DOWN RESPIRATION, IF IT'S --
MITOCHONDRIAL RESPIRATION AND
JUST EVERYTHING IS RUNNING ON
GLYCOLYSIS OR IF IT'S MORE
SOPHISTICATED THAN THAT AND
HAVING SOME ROLE IN OX PHOS FREA
RADICAL, WE DON'T KNOW.
>> WELL, HAPPY TO CHAT AFTER
THIS MEETING.
>> IT'S A LITTLE BIT MORE
EMBARRASSING THAN IT IS ALREADY.
>> SO DAN, MAYBE YOU WANT TO
COMMENT ON HOW YOU SEE CELL
THERAPEUTICS MOVING FORWARD FROM
HERE, WHERE WE ARE?
>> WELL, I THINK WHAT MOST
PEOPLE DON'T REALIZE IS SOME OF
THE TRIALS OF THE MSC BASED CELL
THERAPY FOR COVID ARDS WERE
SUCCESSFUL.
AND WHAT WE NEED TO DO IS
UNDERSTAND BETTER HOW THEY WERE
SUCCESSFUL AND THEN LEVERAGE
THAT TO A BROADER RANGE OF
INDICATIONS.
AND AGAIN TO COME BACK TO THE
PATIENT INFLAMMATORY PHENOTYPE
AS BEING A GUIDE.
SO WHAT WOULD YOU DO THERE?
YOU WOULD INCORPORATE POINT OF
CARE ASSESSMENTS, SO SOMEONE
COMES IN WITH ACUTE LUNG INJURY
OR SEPSIS, SEPTIC SHOCK OR ARDS
OR WHAT HAVE YOU, YOU DO A QUICK
DIAGNOSTIC AT THE BEDSIDE AND
SAY YOU ARE SOMEWHAT WHO IS MORE
LAKELY THAN NOT TO RESPOND TO AN
MSC-BASED SYSTEMIC
ADMINISTRATION, AND THEN GO FROM
THERE.
BUT IT ALSO HIGH LATE A DICTUM
THAT SAYS EVERY CLINICAL
INVESTIGATION NEEDS TO HAVE AS
MANY MECHANISTIC INVESTIGATIVE
ARMS AS POSSIBLE.
SO AS WE DESIGN THE FUTURE
TRIALS, WE NEED TO BE SMARTER
ABOUT BEING ABLE TO PULL MORE
INFORMATION OUT OF THE TRIALS
OVER AND ABOVE THE CLINICAL
OUTCOMES DATA.
I WANT TO ACTUALLY GO OFF A
LITTLE BIT ON WHAT FRANK WAS
SAYING, THOUGH, MITOCHONDRIAL
THERAPIES ARE A NEXT WAVE, IF
YOU WILL.
AND SO IT ONE THING TO
ADMINISTER CELLS.
IT ANOTHER THING TO ADMINISTER
THE EXTRACELLULAR VESICLES OR
THE SECRETOME FROM THE CELLS,
AND THERE'S A SUBSTANTIAL
LITERATURE THAT THOSE
PARTICULARLY EVs MIGHT BE AS
EFFICACIOUS AS THE PARENT CELL,
BUT AGAIN WE HAVE TO UNDERSTAND
HOW THEY'RE WORKING.
BUT THEN THE MITOCHONDRIA, I
MEAN, THERE'S A GROWING
LITERATURE THAT MITOCHONDRIAL
ADMINISTRATION, AND IF YOU THINK
ABOUT ARDS AND SOME OF THE
PATHOGENESIS IS ATP DEPLETION IN
THE TYPE 2 CELLS LEADING TO
SURFACTANT PRODUCTION DEPLETION,
ET CETERA.
AND HOW MITOCHONDRIAL TRANSFER
CAN POTENTIALLY RESCUE THAT.
IT'S ONE PIECE OF THE JIGSAW
PUZZLE, BUT IT AN INTRIGUING
PIECE THAT WHEN YOU'RE THINKING
OF FUTURE DIRECTIONS, IS ONE TO
INCORPORATE INTO THE OVERALL
PROGENITOR MINDSET, IF YOU WILL.
>> SO DAN, CAN I ASK A QUESTION?
IT'S ALWAYS ANNOYED ME THAT
THERE HAVE BEEN SO MANY MSC
STUDIES.
AS YOU SAY, THEY DON'T
INCORPORATE BUT THEY HAVE
ANTI-INFLAMMATORY AND REPARATIVE
FUNCTIONS.
AND OF COURSE THE TIMING IS ALL
FOR AUTOLOGOUS TRANSPLANTS, YOU
KNOW, WE CAN SEE HOW EASY THAT
IS AND THOSE DO INCORPORATE AT
LEAST IN MICE, SO WE ALWAYS
THOUGHT THAT WOULD BE THE IDEAL
THING, BUT OF COURSE THE TIMING
IS SO BAD, PATIENTS SHOW UP IN
THE ICU WITH ARDS, AND YOU
CAN'T -- YOU CAN'T GROW THEM UP
FAST ENOUGH TO DO.
>> COUPLE THAT, FRANK, WITH THE
LIKELIHOOD, AND THERE'S DATA IN
SOME CONDITIONS THAT MSCs
OBTAINED FROM AN ACUTE
INFLAMMATORY INJURY ARE NOT
NECESSARILY PROPERLY
FUNCTIONING, SO IT'S A DOUBLE
WHAMMY.
YOU CAN'T GET THEM, YOU CAN'T
EXPAND THEM, YOU CAN'T GET
ENOUGH, AND THEY MAY NOT WORK,
IF YOU WILL.
THIS GETS YOU BACK TO
ALLOGENEIC.
BUT ALLOGENEIC ARE STILL ALER JE
ALLOGENEIC.
IT WILL STILL PROVOKE A HOST
RESPONSE FAIRLY QUICKLY ONCE THE
EVENT, SPECIFICALLY WITH
SYSTEMIC ADMINISTRATION.
THERE'S A SKOLL SCHOOL OF THOUGT
IT'S NOT THE CELLS THAT ARE
ACTUALLY DOING ANYTHING,
ALTHOUGH THEY CAN DO THAT LONG
LIST OF THINGS, BUT IT'S THE
HOST-IMMUNE RESPONSE THAT'S
REALLY DRIVING THE HOPEFULLY
BENEFICIAL RESPONSE.
>> I SEE, I SEE.
THEN THE OTHER THING WAS, PEOPLE
WERE TALKING ABOUT -- IN THE
DAWN OF IPSCs, PEOPLE WERE
TALKING ABOUT WELL, LET'S GET A
WHOLE BUNCH OF IPSCs GOING
FROM ALL THE ALLO TYPES,
EVERYONE IS MHC, TO TRY TO FOCUS
ON AS MANY AS POSSIBLE.
>> IT'S KIND OF LIKE CORD BLOOD
BLANKING.
BANKING.
THERE'S A WHOLE BUNCH OF HYPE
BUT WHAT DOES THAT EVER -- DO
NOT MISUNDERSTAND, I'M NOT
DISMISSING OR DOWNPLAYING, BUT
WHERE IS THE PROOF, WHERE IS THE
PUDDING HERE, IF YOU WILL?
>> RIGHT.
OKAY.
>> SO DAN, IF THESE CELLS ARE
NOT IMMUNE PRIVILEGED -- CAN YOU
HEAR ME?
>> YES.
>> IF THESE CELLS ARE NOT IMMUNE
PRIVILEGED AND ACT LIKE
ALLOGENEIC CELLS AND ARE
ULTIMATELY CLEARED, THAT'S
PRESUMABLY THEN HYPED UP THE
IMMUNE SYSTEM FOR THOSE CELLS.
WHAT HAPPENS -- IS THERE ANY
CONCERN OR ANY EXPERIENCE WITH
THE SECOND OR THIRD INFUSION OF
MESENCHYMAL STEM CELLS?
>> THAT'S A GREAT QUESTION, HAL.
IT GOES BACK TO THE QUESTION OF
DOSING REGIMEN, BECAUSE WHEN YOU
LOOK AT THE PRE-CLINICAL DATA,
MOST OF THOSE ARE SINGLE
ADMINISTRATION, IF YOU WILL.
BUT WHEN YOU GET INTO THE
CLINICAL TRIALS, SOME OF THEM
HAVE TWO OR THREE CELL
ADMINISTRATIONS SE SEPARATED
GENERALLY OVER A COUPLE OF DAYS
IF YOU WILL, BUT THERE'S BEEN NO
DATA FORTHCOMING AS TO WHETHER
OR NOT ONE HAS DEVELOPED LACK OF
TOLERANCE OR SOME SORT OF MORE
ROBUST IMMUNE RESPONSE.
>> LIKE HAN F ANAPHYLAXIS?
>> NO, NO, NO, HOPEFULLY THAT
CAME THROUGH IN THE CHAT
RESPONSE TO AMANDA HILL AND ALL
THAT.
THE ONE THING THAT'S COME OUT OF
ALL THESE TRIALS IS SAFETY.
EVEN IN CRITICALLY ILL ARDS
PATIENTS ON THE VENTILATOR, ON
ECMO, THAT CLOSE TO DEATH AND
ALL THAT, NOBODY HAS FOUND ANY
ADVERSE EFFECT OF GIVING THE
MSC.
SO IN IMMUNE RESPONSE, THERE'S
THIS HUGE UMBRELLA.
WHAT DOES THAT MEAN?
SO IF IT MEANS THAT ONE IS
REVVING UP MACROPHAGES BECAUSE
THEY'RBECAUSE -- BUT THEY'RE THN
SECRETE IL10, I'M JUST GIVING
YOU SOME BROAD STROKE EXAMPLES,
AND THAT DOES SOMETHING TO RESET
THE HYPERINFLAMMATORY RESPONSE
THAT'S DETRIMENTAL IN ARDS, THEN
THAT'S A WIN-WIN SITUATION.
BUT HAL, SO FAR NOBODY HAS
DROPPED DEAD.
NOBODY HAS HAD ANAPHYLACTIC
RESPONSE.
>> IS ANYONE ENGINEERING THESE
MSCs?
FOR INSTANCE, FOR SECRETE IL10
OR SOME OTHER --
>> SO THE ONE EXAMPLE I
MENTIONED IS STUDIES,
ENGINEERING MSCs TO SECRETE
TRAIL, TNF ALPHA RECEPTOR --
WITH TARGETING AN ANTITUMOR
AGENT.
SO THERE'S BEEN LESS WORK IN
TERMS OF MSCs SECRETE IL10,
BUT THERE HAS BEEN SOME WORK OR
OTHER THINGS WITH IL10 AS YOU
MENTIONED IN TRYING TO PRESERVE
LUNGS FOR TRANSPLANT.
SO A DONOR LUNG GETS TAKEN OUT,
CAN BE PUT ON AN EXTRA VASCULAR
LUNG PERFUSION DEVICE,
UNIVERSITY OF TORONTO HAS REALLY
PIONEERED THIS, AND MSCs,
ENGINEERING TO SECRETE IL10 THAT
POTENTIALLY COULD BE UTILIZED
INTO THAT CIRCUIT TO FURTHER
INCREASE THE VIABILITY OF THE
LUNGS.
IN A PARALLEL TYPE OF ENDEAVOR,
NOT WITH MSCs BUT WITH
ENDOTHELIAL PROGENITOR CELLS,
DUNCAN STEWART AT THE OTTAWA
HEALTH RESEARCH INSTITUTE HAS
ENGINEERED THE EPCs TO
OVEREXPERIENCE ANDRO POIETIN
WITH THE IDEA THAT THIS WOULD BE
BENEFICIAL FOR HYPERTENSION, HAS
HAD A CLINICAL TRIAL GOING ON
THAT AS WELL.
SO SHORT ANSWER, YES, IT'S AN
OPEN FIELD BUT LESS WELL
TRAVELED AT PRESENT.
>> THANK YOU.
>> I'M TOLD WE NEED TO WRAP UP
FOR DINNER -- FOR LUNCH.
>> NO, FIVE MORE MINUTES.
WE CAN TAKE A 20-MINUTE BREAK.
CONTINUE.
FIVE MORE MINUTES.
THANK YOU.
>> IN TERMS OF ENDOTHELIAL
CELLS, WEILL CORNELL IS ALSO
ENGINEERING MICROVASCULAR
ENDOTHELIAL CELLS AS A
THERAPEUTIC.
SO I THINK THERE'S A LOT OF
OPPORTUNITY THERE.
>> I THINK SO.
BUT IT RAISES ANOTHER LEVEL OF
COMPLEXITY WITH THE FDA.
AND EUROPEAN UNION, FOR EXAMPLE,
AND ALL THAT.
SO FOR THE MSC, THE UMBRELLA
TERM MSC, THE REGIONAL
HEAVY-DUTY INVESTIGATIONS 10
YEARS AGO, FOR EXAMPLE, WERE IN
GRAFT VERSUS HOST DEEDS, AND THE
ENTITIES DOING THAT, EITHER
ACADEMIC OR INDUSTRY, HAD TO
JUMP THROUGH MANY, MANY, MANY
HOOPS WITH THE FDA FOR SAFETY
INDICATIONS IN PARTICULAR.
BUT BY AND LARGE, THE MSC
EXPERIENCE OF SAFETY, THE TRACK
RECORD OF SAFETY, IS SUCH THAT
IT'S LESS WORK TO PUT AN MSC
TRIAL FOR FDA APPROVAL.
ONCE YOU START ENGINEERING, IT'S
A WHOLE CAN OF WORMS AND YOU
HAVE TO REINVENT THE WHEEL
BECAUSE IT'S A NEW BEAST.
AND SO THAT'S WHETHER OR NOT ONE
AS AN ENTITY WANTS TO UNDERTAKE
EVERYTHING INVOLVED IN THAT IS A
SERIOUS DECISION TO MAKE.
AND SO THE PRE-CLINICAL DATA,
THE RATIONALE HAS GOT TO BE SO
SOLID AND CONVINCING FOR
SOMEBODY TO BE ABLE TO DO THAT.
>> I THINK WE CAN WRAP UP FOR
DINNER NOW.
>> SO EVERYBODY COME TO VERMONT
NEXT SUMMER FOR THAT CONFERENCE.
>> CAN I MAYBE -- JUST ONE
MINUTE BEFORE WE WRAP UP?
>> YES, WELLINGTON.
>> AGAIN TO FRANK, BECAUSE I'M
VERY FASCINATED BY WHAT HE HAD
DESCRIBED AND PUBLISHED ON THIS.
FRANK, WHEN YOU TALK ABOUT THE
FETAL ORIGIN OF THE CELLS, I
SUPPOSE THIS MUST BE IN HUMANS
BECAUSE IN THE MOUSE, I CAN'T
REMEMBER WHEN I READ YOUR PAPER
WHETHER YOU HAVE DESCRIBED THEM
IN THE MOUSE AS WELL,
PARTICULARLY IN -- AIRWAYS.
>> THIS IS ALL IN HUMAN.
WHEN WE WERE IN BOSTON, WE
RECEIVED A NUMBER FROM CHRIS KRO
KROMM AT THE BRIGHAM, AND IT'S
NOT TRIVIAL TO -- WE LIVE IN
TEXAS NOW.
WE COULD BE -- FOR THIS KIND OF
STUFF, SO WE DID ALL THE
ANALYSIS AND CLONING FROM --
>> BUT FRANK, THOSE CELLS, DO
YOU -- IN RESPONSE SOME OF THOSE
SIGNALS THAT WE HAVE BEEN
TALKING ABOUT, FOR EXAMPLE, --
ACID, FOR EXAMPLE, NOTCH
SIGNALING, FOR EXAMPLE, AND
FORGIVE ME, I MAY HAVE MISSED,
DID YOU NOTICE SOMETHING THAT
ACTUALLY MODULATE THIS EXPANSION
THAT HAS TO DO WITH SOME OF THIS
MORE CANONICAL SIGNALS, TGF
BETA, NOTCH --
>> WELLINGTON, ALL THOSE THINGS
ARE IN THE COMPONENTS THAT --
HAS PUT TOGETHER IN THE MEDIA.
THE MEDIA ARE COMPLEX.
>> I SEE.
>> TO MAINTAIN THEM AS STEM
CELLS.
BUT MANY OF THE PATHWAYS YOU
JUST MENTIONED ARE PART AND
PARCEL, BUT SHE'S PUBLISHED
THAT.
IF YOU ASK ME, I DON'T THINK I
COULD LIST THEM OFF FOR YOU, BUT
SHE COULD.
>> YEP, I UNDERSTAND.
WHAT I WAS JUST WANTING TO TRY
TO FIGURE OUT IS WHEN YOU
INHIBIT THE THINGS, IF ANY OF
THESE PATHWAYS INHIBIT YOU HAVE
A PREFERENTIAL EXPANSION OF A
PARTICULAR CLONE VERSUS ANOTHER,
AND MORE ABOUT RESPONSE --
>> IT'S A GOOD QUESTION, AND
THIS HAS COME UP RECENTLY, SO
ONE THING WAS DOING WITH THIS IS
CLONING OUT THESE DIFFERENT
CELLS IN PANCREATIC CANCER.
SO IT SOUNDS WEIRD, BUT IT WAS
VERY CLEAR THAT THERE'S
HETEROGENEITY IN THOSE
INDIVIDUAL TUMOR PATIENTS OF
THEIR STEM CELLS.
SHE WAS NOT ABLE TO GET THE
BASAL VERSION OF PANCREATIC
CANCER.
SHE HAD SWITCHED TO ANOTHER
MEDIA AND WAS ABLE TO GET THAT.
SO THAT MAKES IT -- WE GET WHAT
WE GET, RIGHT?
SO ARE WE MISSING THINGS?
AND I THINK THAT'S A VERY REAL
POSSIBILITY, THAT IF WE HAD
SLIGHTLY DIFFERENT MEDIAS, WOULD
WE CAPTURE MORE.
BUT WE ARE CAPTURING THEM IN A
RECURRENT WAY.
SO FOR INSTANCE, COPD, ALL THE
CLONES THAT WE SEE IN COPD ARE
IN CYSTIC FIBROSIS.
CYSTIC FIBROSIS HAS TWO
ADDITIONAL ONES THAT ARE UNIQUE
TO IT.
>> I SEE.
>> BUT IT IS -- YOU GET WHAT YOU
GET.
IF WE HAD UNLIMITED PATIENTS, WE
COULD MAKE A WHOLE SET OF MEDIAS
AND TRY TO CLONE OUT MORE.
BUT YOU KNOW --
>> WE HAVE AMANDA HILLARYLY
QUHILL REALLYQUICKLY BEFORE THE.
>> HI.
THANK YOU VERY MUCH.
DANIEL, THANK YOU SO MUCH FOR
THE DISCUSSION.
I'M A VETERINARIAN, AND IN ORDER
TO HAVE A BETTER CLINICAL OUT
COME IN ALLOGENIC TRANSPLANTS,
IF THE PATIENT NEEDS MORE THAN
ONE APPLICATION WE USUALLY
CHANGE THE DONOR AND WE OBSERVE
THE CLINICAL OUTCOME IS BETTER
AS COMPARED TO USING THE SAME
DONOR.
IS THERE ANY STUDIES IN HUMANS
THAT SHOW THAT?
>> AMANDA, IF THE QUESTION IS
TOWARDS ME, WHAT A FASCINATING
IDEA.
AND THE ANSWER IS, NO, NONE THAT
I'M AWARE.
WITBUT AGAIN, THERE ARE ONLY A
LIMITED NUMBER OF STUDIES WITH
MULTIPLE ADMINISTRATIONS, IF YOU
WILL.
BY AND LARGE, IT'S A SINGLE
ADMINISTRATION.
BUT THEN BACK TO HAL'S POINT, SO
FAR IN THOSE MULTIPLE
ADMINISTRATION STUDIES, THERE
HASN'T BEEN ANY OBVIOUS DOWNSIDE
TO USING THE SAME SOURCE.
>> OKAY, GREAT.
THANK YOU.
>> GOOD QUESTION.
REALLY INTERESTING QUESTION.
>> WELL, I THINK WE CAN CONCLUDE
OUR DISCUSSION.
THANK YOU, EVERYBODY, FOR YOUR
PARTICIPATION, AND WHAT A LIVELY
DISCUSSION AND WONDERFUL TALKS.
>> WELL, THANK YOU BOTH FOR
MODERATING, AND FOR DOING THIS.
GREAT.
>> THANK YOU ALL.
>> THANKS, EVERYONE.
SO I HAVE THE PLEASURE HERE TO
KICK OFF THIS SESSION, TOGETHER
WITH AMANDA JAMIESON.
AMANDA IS GOING TO GIVE A LITTLE
BIT OF A BRIEF OVERVIEW TO THE
SESSION AND THEN I THINK GOING
TO GO AHEAD AND INTRODUCE OUR
FIRST SPEAKER.
>> THANKS, JOSE.
SO THE OVERVIEW OF THIS SESSION
IS TO REALLY KIND OF PULL
INTEREST LOTS OF DIFFERENT
SYSTEMS BECAUSE THE LUNG IS NOT
AN ISOLATED ENVIRONMENT, CLEARLY
IT PART OF THE WHOLE BODY, AND
SO REALLY TO GET INSIGHTS FROM
OTHER FIELDS TO UNDERSTAND
MECHANISMS OF LUNG PROGENITOR
RESILIENCE.
THEN OF COURSE AS WE'VE BEEN
DOING FOR THESE LAST TWO DAYS IS
REALLY IDENTIFY CRITICAL GAPS
AND CHALLENGES AND OPPORTUNITIES
AND SOME NEW KEY RESEARCH
QUESTIONS ON HOW WE CAN PULL
FROM THESE MULTIDISCIPLINARY
AREAS.
SO CAN WE GET STARTED WITH OUR
FIRST SPEAKER, WHO IS HELEN C.
MIRANDA FROM CASEWESTERN
UNIVERSITY.
AS USUAL, WE'LL HAVE THE
DISCUSSION AT THE END, AND YOU
CAN PUT ANY SORT OF QUESTIONS IN
THE CHAT FOR CLARIFICATION
QUESTIONS.
>> HELLO.
CAN YOU ALL HEAR ME WELL?
YES?
OKAY.
>> YES, WE CAN HEAR YOU.
>> THANK YOU.
THANKS SO MUCH FOR THE
INVITATION.
I'VE BEEN THOROUGHLY ENJOYING
THIS MEETING, LEARNING A LOT
FROM DIFFERENT FIELDS BUT IT'S
VERY INTERESTING TO SEE HOW,
FROM DIFFERENT FEALDZ, FIELDS, N
FIND THIS INTERSECTION.
SO TODAY I'M GOING TO TALK ABOUT
IPINTEGRATED STRESS RESPONSE AND
HOW WE IDENTIFY PATHOGENS
ASSOCIATED TO THIS NEUROLOGICAL
DISEASE CALLED ALLS.
ALS.
SO MY LAB FOCUSES ON THE STUDY
OF DIFFERENT TYPES OF
NEUROLOGICAL DISEASES.
WE HAVE A LARGER FOCUS ON
NEUROMUSCULAR DISEASES, BUT WE
HAVE ALSO DABBED IN THE FIELD OF
NEUROPEDIATRIC DISORDERS AS
WELL.
FOR THE PURPOSE OF THIS TALK
TODAY I'M GOING TO FOCUS ON ALS.
WE HAVE BEEN MODELING THIS
DISEASE USING THIS VERY SPECIFIC
SYSTEM THAT I WAS GLAD TO SEE
WAS REFERENCED TODAY AND
YESTERDAY BY DIFFERENT SPEAKERS.
SO WE ONLY DO STEM CELL MODELING
IN THE LAB.
THIS IS OUR EXPERTISE.
WE WORK WITH PEOPLE THAT USE
MOUSE MODELS, WITH HE WORK WITH
PEOPLE WITH FLY MODELS, WE HAVE
TONS OF COLLABORATIONS USING
OTHER MODELS BUT THIS IS OUR
EXPERTISE.
AS MANY SPEAKERS HAVE DISCUSSED
AND EMPHASIZED IN PREVIOUS
TALKS, BEFORE THE ADVENT OF
IPSCs, THE AVAILABLE
APPROACHES TO DIRECTLY STUDY IN
OUR CASE SPECIFICALLY
NEUROLOGICAL DISEASES WAS
POSTMORTEM HUMAN TISSUES,
IMMORTALIZED CELL LINES AND
ANIMAL MODELS AND POSTMORTEM
BRAIN TISSUES NOT ALWAYS
REPRESENT -- OFTEN REPRESENTS
THE END STAGE OF THE DISEASE AND
ARE OFTEN NOT WELL PRESERVED AND
WHILE MORE -- SPECIFICALLY
NEUROGENERATIVE DISEASE
PHENOTYPES.
MOST MOUSE MODELS ARE VERY
LIMITED TO SPECIFIC VARIANTS, SO
WE SELECT THIS MODEL WHERE WE
OBTAIN CELLS FROM PATIENTS, THE
FIBROBLASTS OR BLOOD CELLS, WE
THEN REPROGRAM THE CELLS IN
VITRO TO GENERATE STEM CELLS TO
BEHAVE AS IF THEY WERE EMBRYONIC
STEM CELLS IN THE DISH.
IN OUR LAB WE'RE EXPERT AT
DIFFERENTIATING CELLS INTO
DIFFERENT CELL TYPES SO WE MAKE
PROGENITOR CELLS, WE MAKE
SPECIFICALLY CORTICONEURONS,
ASTROCYTES AND OTHER GLIA SUCH
AS MICROGLIA AS WELL AS SKELETAL
MUSCLES.
BUT FOR THE PURPOSE OF THE TALK
TODAY, I'M GOING TO FOCUS ON HOW
WE'RE DIFFERENTIATING THESE
CELLS INTO MOTOR NEURONS.
SO WHAT IS AMYOTROPHIC LATERAL
SCLEROSIS DISEASE I'M GOING TO
FOCUS ON TODAY?
IT'S THE MOST COMMON ADULT ONSET
MOTOR NEURON DISEASE.
ITS PATHOPHYSIOLOGY HAS
INVOLVEMENT OF BOTH UPPER AND
LOWER MOTOR NEURON.
THIS IS TRULY A DEVASTATING
DISEASE.
THE PATIENTS SUFFER IN WHAT WE
CALL THE LOCKING SYNDROME, THEY
ESSENTIALLY LOSE THE ABILITY TO
MOVE, THEY BECOME WHEELCHAIR
BOUND, SOMETIMES BEDRIDDEN, AND
WHILE MOST OF THE PATIENTS DO
NOT SUFFER ANY COGNITIVE
IMPAIRMENT, SO THEY ARE FULLY
AWARE OF THEIR SURROUNDINGS BUT
THEY JUST CAN'T MOVE.
AND UNFORTUNATELY, THE LIFE
EXPECTANCY POST DIAGNOSIS IS
BETWEEN TWO TO FIVE YEARS.
AND IT'S IMPORTANT TO POINT OUT
THAT 90% OF THESE CASES ARE
CONSIDERED SPORADIC, WHERE
THERE'S NO -- PATTERN, BUT 10%
ARE KNOWN TO BE FAMILIAL WHEN WE
IDENTIFY THE DISEASE AND THIS
HAS A PATTERN OF HEREDITARY.
SO TO DATE, WE HAVE OVER 38
VARIANTS IN DIFFERENT GENES THAT
HAVE BEEN ASSOCIATED OR KNOWN TO
BE CAUSATIVE OF DIFFERENT TYPES
OF ALS.
HERE IN THIS GRAPH, WE HAVE IN
BLUE REPRESENTATION OF IMEENS
THATGENESWHICH HAVE BEEN ASSOCIH
ALS, IN RED WE HAVE GENES
ASSOCIATED WITH SPORADIC ALS.
SO THE MAJORITY OF THE GENES
HAVE BEEN IDENTIFIED IN BOTH
POPULATIONS BUT THE ONE I'M
GOING TO FOCUS ON TODAY IS THIS
GENE THAT IS CAUSATIVE OF A RARE
FORM OF ALS CALLED VAPB.
ALL THE WORK I'M GOING TO
DESCRIBE TODAY WAS IN MY LAB BY
MY PH.D. STUDENT, CURRAN LANDRY.
SO WHAT IS VAPB IS THIS HIGHLY
CONSERVED EXPRESSED PROTEIN.
IT'S A MEMBRANE PROTEIN ANCHORED
TO THE MEMBRANE OF THE AR AND IT
HAS THESE THREE DIFFERENT
DOMAINS.
THE TRTHE -- IS THE ONE THAT BIS
DIFFERENT ORGANELLES AND
PROTEINS TO THE MEMBRANE OF THE
AR AND THIS IS THOUGHT TO BE --
EASILY FUNCTION.
SO MUTATIONS THAT HAVE BEEN
ASSOCIATED TO -- MOSTLY
IDENTIFIED IN THAT DOMAIN SO IT
CAUSES A CHANGE FROM A PROLINE
TO A SERINE IN THE POSITION
OF -- 56, FIRST IDENTIFIED IN
THE BRAZILIAN POPULATION.
WE STILL ESTIMATE THE MAJORITY
OF THE PATIENTS WITH THIS
MUTATION ARE FROM BRAZIL BUT
DIFFERENT PATIENTS HAVE BEEN
IDENTIFIED IN OTHER PLACES, IN
OTHER COUNTRIES.
SO AS I MENTIONED, WE USE THESE
INDUCED PLURIPOTENT STEM CELLS
TO MODEL THE DISEASE AND WE'RE
GOING TO FOCUS ON THE MOLECULAR
MECHANISMS THAT ARE ASSOCIATED
WITH THE MUTANT VAPB AND IT'S
CAUSATIVE OF AMMO TROPHIC
LATERAL SCLEROSIS.
TO DO THAT WE ALSO RELY ON
CRISPR ENGINEERING SO THE FIRST
STEP FOR THIS PROJECT WAS TO
ACTUALLY KNOCK OUT VAPB FROM
THESE CELL LINES AND EITHER
EXPRESS THE VAPB WILD TYPE OR
THE MUTANT VAPB, SO WE THEN
GENERATE THIS PANEL THAT WE CALL
THESE ISOGENIC CELL LINES, SO
THEY HAVE ALL THE SAME GENETIC
BACKGROUND WITH THE DIFFERENCE
OF THESE SPECIFIC MUTATION.
SO FOR THIS SPECIFIC DISEASE
WE'RE GOING TO TALK ABOUT HOW
WE'RE GOING TO DIFFERENTIATE
THESE CELLS INTO MOTOR NEURONS.
THE PROTOCOL FOR ALTERNATIVE
DIFFERENTIATION IS REALLY WELL
ESTABLISHED.
WE DIFFERENTIATE THIS USING WHAT
WE CALL THE DUAL -- PROTOCOL.
WE DIFFERENTIATE THESE CELLS
INITIALLY FOR 15 DAYS TO
GENERATE MODERN PRO GENERAL
TORS PROGENITORS.
FURTHER MATURITY CELLS -- THESE
MOTOR NEURONS ARE CHARACTERIZED
BY POSITIVE STAINING OF --
1 AND -- 9.
THIS USES ABOUT 80 TO 90% OF --
NEURONS.
WE THEN -- THEN I'M GOING TO
DESCRIBE HOW WE ASSESS THESE
MOTOR NEURONS WE JUST
DIFFERENTIATED WITH EITHER THE
WILD TYPE VAPB OR THE MUTANT
VAPB SO WE FIRST ASSESS THEM FOR
THEIR FUNCTIONALITY.
IN THIS CASE, WE'RE GOING TO
TALK ABOUT ELECTROPHYSIOLOGY.
WE ALSO LOOK AT THEIR MOLECULAR
PROPERTIES, SO WE ASSESS
THEIR -- WE ALSO ANALYZED THEIR
MITOCHONDRIA AND I'M GOING TO
DESCRIBE THAT TOO, AND THEN AT
THE END, I'M GOING TO TALK ABOUT
HOW WE TIED ER EVERYTHING TOGETR
TO THIS INTEGRATIVE STRESS
RESPONSE MECHANISM THAT WE
IDENTIFIED.
AND ALL OF THE WORK THAT I'M
GOING TO DESCRIBE HERE IS
ACTUALLY AVAILABLE IN --
CURRENTLY AVAILABLE IN
BIOARCHIVES.
SO NEXT, FOR THE FIRST
ASSESSMENT WE WANTED TO LOOK AT
THE FUNCTIONALITY OF THESE MOTOR
NEURONS.
IN ORDER TO DO THAT, WE RELY ON
THIS MACHINE WE HAVE IN THE LAB
CALLED THE MYO STROBE MACHINE
FROM AXE YON.
THIS SYSTEM RELIES ON THESE
ARRAY OF ELECTRODES IN THE
BOTTOM OF WELLS IN A MOTOR WELL
PLATE, AND WE JUST PLATE THE
NEURONS ON TOP OF THESE
ELECTRODES AND THE ELECTRODES
PICK UP THESE CHANGES IN ION
CONCENTRATION AROUND THEM, AND
THAT'S A PROXY FOR THEIR FIRING
PROPERTIES.
SO HERE I BROUGHT TWO OF MANY OF
THE OUTPUTS THAT THIS MACHINE
GIVES US, BUT SHOW BUT TWO OF M
GOING TO SHOW HERE ARE
REPRESENTATIVE OF THESE LOWER OR
DECREASED NEURONAL -- RATE WE
SAW IN THESE NEURONS.
SO WE SAW STATISTICAL
SIGNIFICANCE IN THE WAY THAT --
FIRING RATE AS WELL AS NEURONAL
OUTBURSTS.
SO SINCE THEIR FUNCTION WAS
CLEARLY DIFFERENT BETWEEN THE
TWO, SO WE HAVE THESE
ELECTROPHYSIOLOGICAL
DYSFUNCTION, WE THEN ASK WHAT
ARE THE MOLECULAR MECHANISMS
THAT COULD LEAD -- LEADS THIS
SPECIFIC MUTATION TO THESE
ELECTROPHYSIOLOGICAL
DYSFUNCTION.
IN ORDER TO DO THAT, WE WANTED
TO ASSESS THE VAPB BINDING
PARTNERS IN THE CONTEXT OF THE
MUTANT VERSUS THE WILD TYPE.
I DIDN'T MENTION THAT IN THE
BEGINNING, HOWEVER, THIS WAS
PRECISELY WHY WE DECIDED TO
CRISPR KNOCKOUT INITIALLY AND
THEN EXPRESS THE WILD TYPE OR
THE MUTANT BECAUSE THE PATIENTS
ACTUALLY ARE HETEROZYGOUS FOR
THE DEEDS SO THE PATIENT LINES
HAVE BOTH WILD TYPE AND MUTANT
VAVAPB.
WE REALLY WANTED TO SEPARATE
WHAT ARE THE PROTEINS BINDING
SPECIFICALLY TO THE MUTANT OR
THE WILD TYPE AND COMPARE THE
TWO, AND WHEN WE PERFORMED THIS
COMPARISON, WE NOTICED THAT
THERE WAS A DECREASE IN THE
NUMBER OF BINDING PARTNERS IN
THE MUTANT LINES COMPARED TO THE
WILD TYPE THAT WERE ASSOCIATED
TO THE MITOCHONDRIA.
THIS WAS NOT ENTIRELY TOO
SURPRISING.
THERE WAS ONE SPECIFIC PROTEIN
CALLED PTPIP51, A MITOCHONDRIAL
PROTEIN THAT HAS BEEN SHOWN TO
BIND TO VAPB, IT HAS BEEN SHOWN
TO PLAY IMPORTANT ROLES BETWEEN
THE MITOCHONDRIA AND THE AR, SO
WE VALIDATE THAT IN OUR
FINDINGS, IN OUR -- IN THE
CONTEXT OF THESE MOTOR NEURONS.
AND WE ALSO SEE A DECREASE
INTERACTION BETWEEN -- MUTANT
AND -- COMPARED TO THE WILD TY
TYPE.
SO AS I MENTIONED, THESE
PROTEINS SPECIFICALLY ENSURES
THESE CONNECTION OR THESE
BINDING BETWEEN THE MITOCHONDRIA
AND THE.
A R.
SO WE THEN WANTED TO ASK IF WE
SEE LOSS OF BINDING PARTNERS TO
THE MITOCHONDRIA, WHAT WAS
HAPPENING TO THIS POINT OF
CONTACT BETWEEN THE ER AND
MITOCHONDRIA.
WE PERFORMED THIS ASSESSMENT
THROUGH EM AND WE DO SEE
STATISTICALLY SIGNIFICANT
DECREASE IN THESE ER
MITOCHONDRIAL CONTACT, AND WE
PERFORM THESE IN TWO TIME
POINTS.
THE FIRST ONE WAS -- 5 BECAUSE
WE WANTED A TIME POINT THAT
WOULD REPLICATE THE MATURATION
OF THE NEURON, SO THIS WAS THE
TIME POINT WHERE THE NEURONS
WERE CONSIDERED MATURE.
HOWEVER, THIS IS -- DISEASE SO
WE ALSO WANTED A VERSION OF AN
AGED MODERN NEURONS -- 60 OF
DIFFERENTIATION.
SO FOR THESE SPECIFIC MOLECULAR
PHENOTYPE, WE DIDN'T SEE ANY
DIFFERENCE.
WE SAW THE SIMILAR RESULTS
BETWEEN THE MATURE MOTOR NEURONS
AS WELL AS THE AGED MOTOR
NEURONS, HOWEVER, WHEN WE
DECIDED TO ASSESS THE FUNCTION
OF THE MITOCHONDRIA, WE DID THAT
BY PERFORMING A JC1 ASSAY, VERY
STANDARD MITOCHONDRIA MEMBRANE
POTENTIAL ASSESSMENT.
WE DID NOT SEE ANY CHANGES
INITIALLY WHEN THE NEURONS WERE
MATURE BUT WE SAW STATISTICALLY
SIGNIFICANT DECREASE IN MEMBRANE
POTENTIAL ON DAY 60 WHEN THESE
NEURONS ARE AGED WHICH WAS ALSO
VERY INTERESTING.
SO THEN WE DECIDED TO ASK THE
QUESTION WHAT WOULD BE THE
RESPONSE INTRACELLULARLY TO THIS
MITOCHONDRIAL DYSFUNCTION.
BASED ON THE KNOWLEDGE THAT THIS
CONTACT BETWEEN THE MITOCHONDRIA
AND ER IS VERY IMPORTANT FOR
HOMEOSHOMEOSTASIS, CALCIUM HAND,
AS WELL AS THE PRELIMINARY DATA
I'M NOT GOING TO SHOW TODAY, WE
DECIDED TO LOOK AT TRANSLATION,
A VERY ENERGETIC, INTENSIVE
PROCESS.
SO WE DECIDED TO LOOK INITIA
INITIALLY -- AND SAW INCREASE IN
PHOSPHORYLATION INDICATING THAT
MRNA TRANSLATION OR PROTEIN
SYNTHESIS MIGHT BE DEPLETED --
SO THEN WE ASSESS THE
TRANSLATION ITSELF BY USING
THE -- ASSAY THAT RELIES ON THE
POST OF PURE MY SEEN, IS
SUPPOSED TO BE INCORPORATED TO A
NASCENT PROTEIN AND THAT GIVES
YOU A PROXY TO HOW TRANSLATION
RATES ARE AT A GIVEN TIME.
OH IN SO IN THIS CASE WE THEN ET
THE PROTEIN, AND IN THE CONTEXT
OF THE WILD TYPE VERSUS THE
MUTANT, WE SEE CLEARLY A
DECREASE IN THIS PROTEIN,
INDICATING A DECREASE OR
SHUTDOWN IN MRNA TRANSLATION.
BUT THEN HOW DO WE TRY THESE
TRANSLATION DEFECTS TO THE
MITOCHONDRIAL DYSFUNCTION THAT
WE'VE SEEN BEFORE?
SO RECENTLY LITERATURE HAVE
POINTED TO THIS PROTEIN CALLED
DELE1, THE END OF A PROCESS
CALLED INTEGRATED STRESS
RESPONSE WHICH IN THIS CASE
WOULD BE THE PROTEIN SYNTHESIS.
SO MITOCHONDRIAL DYSFUNCTION
WOULD ACTIVATE DELE1 AND WOULD
CLEAVE IT INTO DELE1S THAT WOULD
THEN BE TRANSPORTED FROM THE
MITOCHONDRIA TO THE -- TO
ACTIVATE HRE, AND ACTIVATION OF
HRE -- ACTIVATE THIS PROCESS
CALLED INTEGRATED STRESS
RESPONSE THAT CULMINATES WITH
PHOSPHORYLATION TO -- ALPHA,
DECREASE OF PROTEIN SYNTHESIS
AND ACTIVATION OF ATF4.
SO IN THIS CASE, WE DECIDED TO
LOOK AT DELE1.
WE DO SEE AN INCREASE IN THE
LEVELS OF DELE1 S IN THE MOTOR
NEURONS COMPARED TO ITS CONTROL.
THEN THIS BECAME OUR PROPOSED
MECHANISM.
SO THERE'S A BREAK IN THE
CONTACT BETWEEN THE MITOCHONDRIA
AND ER SHOWN BY EM.
THIS WOULD THEN ACTIVATE DELE1
CLEAVAGE WHICH WOULD ACTIVATE
INTO A STRESS RESPONSE.
IT WILL BE RESPONSIBLE FOR ALL
THESE PHENOTYPES THAT I HAVE
DESCRIBED.
SO I HAVE SHOWN HERE THAT WE SEE
THIS INCREASE IN DELE1S.
I ALSO SHOW WE SEE AN INCREASE
IN PHOSPHO -- 2 ALPHA AS WELL AS
AN ACCOMPANIED DECREASE IN --
TRANSLATION IN THIS CASE.
SO THE QUESTION HERE WAS WHAT
HAPPENS IN WE INHIBIT THIS
PROCESS?
WHAT IF WE INHIBIT THE STRESS
RESPONSE USING THIS INHIBITOR
THAT HAS ALSO BEEN DESCRIBED
YESTERDAY IN HUNT'S TALK.
SO WE DECIDED TO TEST THIS DRUG
TO SEE IF IT COULD INHIBIT THE
PROCESS AND ASK THE QUESTION
COULD THIS DRUG THEN RESCUE ANY
OR ALL OF THESE PHENOTYPES THAT
I HAVE JUST DESCRIBED.
SO INITIALLY WE WANT TO ASSESS
THE INHIBITION OF ISR AND THE
RESCUE, AND THE FIRST WE
ASSESSED WAS THIS DECREASE IN
TRANSLATION BY RE-ASSESSING THE
SUNSET ASSAY, AND WE DO SEE A
COMPLETE RESCUE OF THE
PHENOTYPE, WE ZO SEE AN INCREASF
MRNA TRANSLATION PROTEIN
SYNTHESIS IN THIS CASE.
THIS WAS NOT TOO SURPRISING THIS
IS WHAT THESE DRUGS ARE SUPPOSED
TO DO, THESE ACTIVATORS ARE
SUPPOSED TO BYPASS THE
PHOSPHORYLATION OF THE ACTUAL
ALPHA AND INCREASE OR ACTIVATE
MRNA TRANSLATION.
HOWEVER, WE'VE ALSO SEEN THAT
THE TREATMENT WAS ALSO ABLE TO
RESCUE THE DEFECT WE SAW
INITIALLY.
SO THESE CELLS RESPONDED BY
INCREASING THE MITOCHONDRIAL
POTENTIALS.
SO BECAUSE THIS WAS A MORE
FUNCTIONAL ASSAY, WE THEN ASKED
CAN THAT BE SUFFICIENT TO THEN
RESCUE THE MOST BIOLOGICAL
RELEVANT PHENOTYPE THAT WE HAVE
IDENTIFIED INITIALLY WHICH
WAS --
>> DR. MIRANDA, WE'RE GETTING
OVER TIME.
ARE YOU WRAPPING UP SOON?
>> YES.
LAST SLIDE.
SORRY.
I DIDN'T HEAR THE SOUND.
SORRY.
THE 2 MINUTES.
OKAY.
SO THIS IS -- NEXT.
SO IT DID RESCUE, AND THE NEXT
ONE WILL BE JUST A SUMMARY.
THANK YOU.
>> THANK YOU.
>> THANK YOU SO MUCH.
THE QUESTIONS WILL BE AT THE
END.
>> THANK YOU VERY MUCHMENT I'D
LIKE TO HAND OVER THE PODIUM NOW
TO SAM RAREDON, WHO'S COMING TO
US TODAY FROM THE VASCULAR
BIOLOGY AND THERAPEUTICS
PROGRAM, SO PLEASE, SAM.
>> HI, EVERYONE.
SAM RAREDON, NEW SYSTEM
PROFESSOR AT YALE AND I'M GOING
TO TALK TODAY A LITTLE BIT ABOUT
CELL-TO-CELL CONNECTIVITY
ANALYSIS.
AND I'M GOING TO SPEND THE FIRST
CHUNK OF THE TALK JUST SORT OF
TALKING ABOUT THE DATA STRUCTURE
AND WHAT YOU CAN DO WITH IT AND
THEN THE SECOND HALF OF THE TALK
IS GOING TO BE A VARIETY OF
DIFFERENT APPLICATIONS BOTH IN
LUNG AND SOME OTHER ORGAN
SYSTEMS JUST SO YOU CAN SEE WHAT
YOU CAN DO AND WHAT YOU CAN
STUDY WHEN YOU STUDY
CELL-TO-CELL COMMUNICATION AS
SORT OF A PRIMARY FOCUS.
SO I JUST WANT TO START WITH
THIS SCHEMATIC, WHICH IS TO SORT
OF POINT OUT, IT'S A TRUISM BUT
THAT WHEN WE STUDY CELLS WITHIN
TISSUES, THEY ARE IN VERY
LOCALIZED SPECIALIZED MICRO
ENVIRONMENTS, AND WE OFTEN WILL
STUDY CELL BIOLOGY AND WE'LL
THINK ABOUT INTRINSIC EFFECTS OF
WHAT A CELL CAN AND CANNOT DO
AND HOW IT MIGHT CHANGE OR NOT
CHANGE AND DIFFERENT PROCESSES,
BUT THERE'S A WHOLE 'NOTHER
LAYER OF INFORMATION THAT WE ARE
SORT OF CUTTING OUT IF WE JUST
STUDY THE CELLS AS INDIVIDUAL
SYSTEMS.
AND THAT'S THE INCOMING
SIGNALING AND THE INFORMATION
THAT'S BEING PROVIDED TO THEM
LOCALLY, WHETHER OR NOT THEY ARE
HEARING IT, AND THE OUTGOING
SIGNALING WHICH IS THE
INFORMATION THERE THEN PROVIDING
TO THEIR LOCAL CONTEXT, WHICH
THEN GOES OUT TO THE OTHER CELLS
NEARBY.
AND WE REALLY STARTED
THINKING -- THIS DIAGRAM, I'VE
STARTED THINKING MORE AND MORE
ABOUT THIS BECAUSE IT'S ACTUALLY
A REALLY NICE WAY OF LOOKING AT
IT BECAUSE IT ALWAYS TREATS
EVERY INDIVIDUAL CELL WITHIN A
TISSUE AS A SINGLE DEDICATED
COMPUTE UNIT THAT'S TAKING IN
INFORMATION, PROCESSING IT AND
SPITTING DIFFERENT INFORMATION
BACK OUT AND ADAPTING EACH TIME
STEP, IT'S LIKE A LITTLE STATE
MACHINE THAT IS CHANGING OVER
TIME.
SO THIS IS SORT OF A FRAMEWORK
FOR THE WHOLE TALK, AND I THINK
SHOWS WHY THE SIGNALING
INFORMATION ITSELF CAN ACTUALLY
BE A VERY INTERESTING TARGET OF
STUDY.
SO THIS IS SORT OF ANOTHER
FRAMEWORK WHICH IS THAT WHEN WE
TALK ABOUT THINGS LIKE TISSUE
BEHAVIOR, MEANING HOW IT
RESPONDS TO AN INJURY OR HOW IT
BECOMES INFLAMED OR HOW IT
REGENERATES OR TISSUE MORPHOLOGY
IN TERMS OF HOW CELLS SELF
ASSEMBLE AND MIGRATE AND HOW
THEY SORT OF FORM THE STRUCTURE
OF TISSUES, AS WELL AS THINGS
LIKE TISSUE STABILITY WHICH I
THINK IS VERY APT FOR THIS
SEMINAR BECAUSE WHEN PEOPLE ARE
TALKING ABOUT RESILIENCE, THE
WAY MY BRAIN IS PROCESSING THIS
IS THINKING OH, THIS IS ABOUT
SYSTEM STABILITY, HOW MUCH IS A
SYSTEM OF CELLS OR INDIVIDUAL
CELLS WITHIN THAT TISSUE WILLING
TO STAY WITHIN HOMEOSTASIS OR
DEVIATE AND COME BACK.
BUT ALL OF THESE DIFFERENT
CONCEPTS, WHETHER WE'RE TALKING
ABOUT BEHAVIOR, MORPHOLOGY AND
SELF ASSEMBLY OR STABILITY,
THERE'S A ROLE TO BE PLAYED HERE
FOR CELL-TO-CELL COMMUNICATION
AND THERE'S A HUGE AMOUNT OF
INFORMATION PROCESSING THAT'S
GOING ON AT THAT LEVEL THAT
BECOMES VERY IMPORTANT FOR
REGENERATIVE MEDICINE
APPLICATIONS IN PARTICULAR.
LIKE DR. SHI LER TALKED ABOUT
YESTERDAY INFLUENCING TISSUE
STATE, PUSHING LARGE SETS FROM
AN ABERRANT PATHOLOGIC STATE,
THIS SHOULDING SOMEBODY WE
PROBABLY SHOULD BE STUDYING AND
THINKING ABOUT HOW WE MIGHT BE
ABLE TO INFLUENCE.
SO THIS IS ANOTHER VIEW OF THIS
WHICH IS THAT YOU CAN ACTUALLY
LOOK AT CONNECTIVITY DATA AS
NETWORK STRUCTURE, AND ALL OF
THE DATA SORT OF PROCESSING
THINGS I'M GOING TO SHOW TODAY
ARE DERIVED FROM THIS CENTRAL
CONCEPT, WHICH IS REGARDLESS OF
WHAT PROCESS YOU'RE STUDYING,
THERE'S A NETWORK ARCHITECTURE
ON THE CELL-TO-CELL SIGNALING
LEVEL, YOU GO FROM MULTIPLE
CELLS TO MULTIPLE KRIL TYPES,
YOU SEE AN INCREASING DIVERSITY
OF CELL TYPES BUT YOU ALSO SEE
INCREASING DIVERSITY OF
CELL-TO-CELL COMMUNICATION
STYLES THAT ARE PRESENT WITHIN A
TISSUE.
THIS EVENTUALLY OF COURSE LEADS
TO DEVELOPMENT OF HOMEOSTASIS.
WHEN WE SEE INFLAMMATION WHERE
WE MIGHT HAVE EX-TRAF JAITION OF
NEW CELLS INTO THE TISSUES THAT
ARE INFLAMED AND ACTIVATING THAT
TISSUE, WE ALMOST GET A NEW NODE
OR A NEW PRESENCE OF A NEW NODE
WITHIN THESE GRAPHS WHERE WE
HAVE HOMEOSTATIC SYSTEM AND THEN
THERE'S NEW CELLS BROUGHT IN AND
IT CHANGES THE GRAPH STRUCTURE
AND IF THE TISSUE HEALS, IT
COMES BACK TO HOMEOSTASIS.
BUT WE ALSO SEE THESE PATTERNS
IN PROGRESSIVE DISEASE, WHICH
I'VE DONE A LOT OF TRAINING IN
PULMONARY FIBROSIS AND WHAT WE
SEE ESSENTIALLY IS THE GRAPH ON
THE BOTTOM, YOU START TO GET
THESE AB BRANDT C ABERRANT CELLU
EVENTUALLY GET THIS SORT OF
PROGRESSIVE DIVERSION OR LOCKED
IN SYSTEM THAT'S NOT ABLE TO
COME BACK.
SO HOW DO WE STUDY THIS?
THERE'S A NUMBER OF STRENGTHS TO
THIS TYPE OF DATA BUT ONE OF
THEM IS WE'RE MEASURING MANY,
MANY CELLS AT ONCE FROM AN
INDIVIDUAL TIME POINT AND THIS
IS SOMETHING YOU WOULD REALLY
STRUG TOLL DO WITH OTHER
TECHNIQUES.
AT A SINGLE TIME POINT YOU CAN
GET SPATIAL DATA AND YOU GET
INFORMATION FROM AN ENTIRE
SYSTEM, YOU CAN SAY THESE CELLS
EITHER ARE INTERACTING OR HAVE
THE POTENTIAL TO INTERACT.
THIS IS JUST AN EXAMPLE OF A
NATIVE RAT THAT WE'VE GENERATED
WHICH I'LL USE SORT OF FOR SOME
DEMONSTRATIONS.
SO IN THESE DATA, ANYONE WHO'S
STUDYING CELL-TO-CELL SIGNAL
WILLING LOOK AT THE LIGAND AND
RECEPTOR EX-TRETION FOR
DIFFERENT CONNECTIVITY
COMMUNICATION MECHANISMS AND
VERY OFTEN THE PATTERNS IN THESE
DATA LOOK LIKE WHAT I'M SHOWING
HERE, WHICH IS THAT ONE CELL
TYPE WILL EXPRESS A LIGAND, THE
OTHER WILL EXPRESS A RECEPTOR,
AND THE FIRST GENERATION TOOLS
OF WHICH THERE ARE LIKE 40 OR 50
OF THEM NOW, WE BROKE ONE OF
THEM, BUT THERE'S CELL PHONE --
THERE'S A LOT OF THEM.
THEY WILL TEND TO AGGREGATE
INFORMING.
THEY WILL SAY WE'RE GOING TO
TAKE YOUR CELL TYPE LABELS AND
WE'RE GOING TO LOOK JUST AS A
WHOLE, WE'RE GOING TO SAY WHAT'S
THE LIGAND EXPRESSION FOR CELL
TYPE A THAT YOU PUT THOSE
ANNOTATIONS ON AND WHAT'S THE --
THEY'LL GIVE YOU A SINGLE IJ
HERE WHICH IS AN AGGREGATED
SCORED VALUE, AND YOU CAN DO A
LOT WITH THIS.
BUT IT LOSES A TON OF
INFORMATION.
SO THIS IS REALLY HOW WE SHOULD
BE DOING THIS IN ORDER TO
MAXIMIZE THE STATISTICAL POWER
OF THIS DATA.
THIS IS APPLICABLE TO LIKE ALL
SINGLE CELL DATA ANALYSIS, BUT
ONE THING THAT WE CAN DO THAT'S
VERY POWERFUL IS RATHER THAN
AGGREGATING, WE CAN GO IN AND
MAKE ONE TO ONE PAIRS OF A GIVEN
CELL AND ANOTHER CELL AND SAY
WE'RE GOING TO TREAT THESE AS
COMMUNICATING OR AGAIN, IF YOU
HAVE SPATIAL INFORMATION, YOU
ACTUALLY DO THIS WITHIN LOCAL
NEIGHBORHOODS, WHICH IS REALLY
POWERFUL.
AND YOU LOOK AT SINGLE EDGES,
YOU LITERALLY LOOK AT ONE CELL
TO ANOTHER CELL AND I IS A LET'S
CHARACTERIZE THAT EDGE AND IN
THIS CASE, WHAT I'M SHOWING ON
THE SCREEN HERE IS JUST ONE
MECHANISM.
WE COULD SAMPLE A HUNDRED PAIRS
OF CELLS FOR A GIVEN MECHANISM,
LIGAND X TO LIGAND Y -- BUT WE
ACTUALLY HAVE MANY THOUSANDS OF
MECHANISMS IN THESE DATA.
SO THIS IS HOW WE STRUCTURE OUR
CONNECTIVITY DATA, AND THERE'S A
COUPLE DIFFERENT PLATFORMS OUT
THERE THAT DO THIS.
FROM EAR ON, I'M GOING TO SHOW
YOU DATA AND ANALYSIS WITH A
PACKAGE WE WRITE AND MAINTAIN
CALLED NICHES, WE HAD TO FIGURE
OUT AN ACRONYM TO MAKE IT LOOK
PRETTY, BUT THIS IS THE
STRUCTURE OF THE DATA AND ONE
THING THAT I THINK IS REALLY
BEAUTIFUL ABOUT THIS IS THAT WE
FIGURED OUT A WAY TO TAKE
CELL-TO-CELL SIGNALING
INFORMATION AN OR REALLY
CELL-TO-CELL CONNECTIVITY DATA
AND FORMAT IT IN A WAY THAT'S
JUST THE SAME AS SINGLE CELL
DATA, WHERE SINGLE CELL DATA
WOULD HAVE A DIGITAL GENE
EXPRESSION MATRIX MEANING THE
ROWS ARE INDIVIDUAL GENES AND
THE COLUMBUS ARE INDIVIDUAL
CELLS, AND WHAT WE'VE DONE HERE
IS WE'VE CREATED A PARALLEL DATA
FRAMEWORK WHERE THE ROWS HERE
ARE SIGNALING MECHANISMS,
MEANING A LIGAND TO A RECEPTOR,
AND THE COLUMNS HERE ARE NOT
CELLS BUT THEY'RE EDGES, THEY'RE
A PAIR OF SENDING CELL TO A
RECEIVING CELL AND THEN WE
CHARACTERIZE ALL OF THESE EDGES
AND PUT THIS RIGHT BACK INTO
SINGLE CELL PLATFORMS AND DO
SOME REALLY COOL PROCESSING.
ONE THING THAT'S IMPORTANT ABOUT
THIS IS WE CARRY THE METADATA
WITH IT.
SO IF YOU HAVE LOCATION
INFORMATION, SAMPLE PATIENT
IDs, DISEASE, APPLICATION OF A
DRUG, THAT METADATA ALL COMES
OVER AND WE CAN SEE IT FROM OUR
SENDING AND RECEIVING CELLS AND
THAT BECOMES VERY IMPORTANT
LATER AND THIS MAKES US VERY
STATISTICALLY FLEXIBLE.
SO THIS IS JUST SORT OF THE
WORKFLOW THAT WE DO AND FROM
HERE ON, I'M GOING TO SHOW YOU
SOME APPLICATIONS.
SO YOU HAVE AN INPUT, A SINGLE
CELL GENE EXPRESSION MATRIX,
SPATIAL TRANSCRIPTOMICS
INFORMATION.
THERE ARE MANY THAT COME WITH
THE TOOL ALREADY BUT YOU CAN
ALSO DESIGN YOUR OWN AND THAT'S
JUST A LIST OF KNOWN OR
HYPOTHESIZED LIGAND RECEPTOR
MECHANISMS THAT YOU WANT TO
QUERY.
AND THE OUT PUME OUTPUTS FROM TA
CELL-TO-CELL OBJECT AS WELL AS
SOME SORT OF HIGHER DIMENSIONAL
SYSTEM CELL OBJECTS AND THEY
ALLOW YOU TO PUT THIS RIGHT INTO
ANY OTHER KIND OF SINGLE CELL
TOOL AND STUDYING IN PARALLEL
WHILE YOU'RE STUDYING TRANS
SCRIP TOE MIBG, OR THE OTHER
TYPES OF PROTEOMIC DATA.
SO THIS IS WHAT THIS LOOKS LIKE.
THIS IS JUST A DEMONSTRATION ON
A SMALL DEMONSTRATION -- YOU CAN
SEE THESE ARE FOUR DIFFERENT
CELL TYPES COLLECTED FROM THE
ALVEOLAR REGION OF THE LUNG.
THERE ARE 16 CELL TYPE TO CELL
TYPE CROSSES THAT YOU COULD
PREDICT SHOULD SHOW UP FROM THIS
IN PANEL I.
YOU CAN SEE UNDER PANEL J,
THERE'S AN EMBEDDING, NOT OF
CELLULAR INFORMATION BUT OF EDGE
INFORMATION, AND YOU CAN SEE THE
ANNOTATIONS HAVE CARRIED OVER SO
THERE ARE DISTINCT SIGNALING
STYLES OR WHAT WE CALL SIGNALING
ARCHETYPES ASSOCIATED WITH EACH
CELL TYPE TO CELL TYPE CROSS.
YOU CAN THEN RUN CELL
DIFFERENTIAL EXPRESSION AND YOU
CAN CHARACTERIZE EACH ONE OF
THOSE SIGNALING ARCHETYPES BASED
ON THE SIGNALING MECHANISMS
THEY'RE MOST LIKELY TO EMPLOY.
IF YOU LOOK CLOSELY AT THE ROWS,
THEY ARE NOT GENES BUT LIGAND
RECEPTOR PAIRS.
YOU CAN THINK OF THEM AS LIKE A
RADIO FREQUENCY, THAT DIFFERENT
CELL TYPES ARE UNIQUELY TUNED TO
WITHIN THIS GIVEN SYSTEM OF
STUDY.
SO THIS IS AN APPLICATION TO
PULMONARY FIBROSIS DATA FROM DR,
AND THIS DATA IS A VERY DENSE
SLIDE BUT EVERYTHING YOU'RE
SEEING HERE IS CONNECTIVITY
INFORMATION, IT'S NOT CELL
INFORMATION.
AND YOU CAN SEE ON THE LEFT SIDE
OF THE SCREEN THAT IT CARRIES
OVER DISEASE IDENTITY AND
SUBJECT IDENTITY, AND THEN WE
HAVE THESE TWO PIECES OF
INFORMATION, WHICH IS SENDING
CELL CLASS AND I'VE CLASSIFIED
THEM AS EPITHELIUM, ENDOTHELIUM,
AND RECEIVING SELL CLASS, AND IF
WE DON'T GO IN, BUT HIS
EPITHELIAL TO MESENCHYMAL
SIGNALING, JUST COMPUTATIONALLY
WE CAN GET A REALLY BEAUTIFUL
PORTRAIT IN CONTROL TISSUE AND
PULMONARY -- IS THAT WHY WHEN
YOU PULL OUT FEATURES YOU CAN
SEE THINGS LIKE CONNECTIVITY VIA
WIN7A AND THROUGH -- ARE REALLY
ENRICHED ALMOST SPECIFIC TO IPF
TISSUE, AND WHEN YOU THEN LOOK
AT THE BOTTOM PANEL HERE WHERE
YOU LOOK AT SENDING AND
RECEIVING CELL TYPE, YOU CAN
ACTUALLY SEE THIS IS SIGNALING
FROM THE ABERRANT BASAL CELLS
WE'VE HEARD SO MUCH ABOUT TO
FIBROBLAST AND MILD FIBROBLAST
CELLS, AND NO OTHER CELLS IN THE
NORMAL HEALTHY LUNG ARE REALLY
TUNED IN THIS WAY WHERE THEY
COULD PASS INFORMATION THROUGH
THESE PATHWAYS BUT WE SEE IT
VERY PROMINENTLY IN PULMONARY
FIBROSIS.
IF YOU'RE INTERESTED, YOU CAN
SEE A PRE-PRINT.
THESE ARE ENGINEERED LUNG
TISSUES WHERE DIFFERENT CELL
COMBINATIONS WITHIN A
BIOENGINEER SCAFFOLD.
ON THE RIGHT WE CAN SEE A QUAD
CULTURE SYSTEM WHERE WE'VE ADDED
IN ALVEOLAR MACROPHAGES.
WHAT'S IMPORTANT FOR THIS TALK
IS THE GRAPH STRUCTURE, WHEN WE
HAVE SINGLE EDGE DATA, WE CAN
ACTUALLY VISUALIZE THOSE IN
THESE U MAP EMBEDDINGS WHICH IS
A REALLY BEAUTIFUL WAY TO LOOK
AAT IT.
YOU CAN SEE HOW THE CONNECTIVITY
STRUCTURE HAS PHYSICALLY
CHANGED.
THIS IS A NICE OFF-TARGET EFFECT
BECAUSE WE'VE ADDED IMMUNE CELLS
BUT THE BIGGEST SEEN WE'RE
SEEING -- FROM EPITHELIUM TO
MESS -- IN THE PRESENCE OF
IMMUNE CELLS.
THIS IS A BUSY SLIDE, GIVEN THAT
I'M TIED ON TIME I'LL FOCUS ON
THE RIGHT SIDE OF THIS.
FOR THOSE INTERESTED IN STUDYING
DYNAMIC PROCESSES, NICHES LENDS
ITSELF REALLY, REALLY WELL TO
THIS BECAUSE YOU GO INTO EACH
ONE OF YOUR SAMPLES, TIME
POINTS, AND YOU CAN CA CALCULATE
CONNECTIVITY FOR EACH ONE AND
THEN MERGE THEM TOGETHER AND
LOOK FOR TRENDS, AND YOU CAN SEE
ACTUALLY CONNECTIVITY CURVES IN
PARTICULAR IN PANEL D, WINT4 --
YOU CAN ALSO SEE THE CELLS THAT
ARE ASSOCIATED WITH THAT CHANGE
AND IN THIS CASE, THE MESS THEEL
YELL TO SORT OF ALVEOLAR
PROGENITOR CELL TUNING IS REALLY
DYNAMIC IN THIS SYSTEM.
THIS IS A NICE -- I THINK
THERE'S A NUMBER OF PEOPLE AND I
THINK WE'RE HEARING A LOT ABOUT
THE DIFFICULTY OF TRANSLATING
ACROSS SPECIES AND THIS IS A
VERY REAL DIFFICULT HARD
PROBLEM.
I'VE GOT A LOT OF COLLEAGUES WHO
ARE ALSO STUDYING THIS, AND OR
THOLG IS NOT ALWAYS CLEAR
BETWEEN HUMANS AND MICE AND RATS
AND FERRETS AND MONKEYS AND
PIGS.
THEY'VE GOT DIFFERENT GENOMES
AND THOSE GENES ARE DISTRIBUTED
IN DIFFERENT WAYS WITHIN THE
DIFFERENT CELLS.
SO IT CAN BE A CHALLENGE.
WE HAVE STARTED TO SEE THAT
CONNECTIVITY STRUCTURE, HOWEVER,
IS A REALLY NICE WAY TO LOOK AT
GENETIC OR THOLG AND CELL TYPE
OR THOLG AND THIS IS JUST AN
EXAMPLE OF SOME DATA FROM RAT
LUNG, WHICH AGAIN I'VE GROUPED
INTO -- AND THEY'VE BEEN
CLUSTERED INDEPENDENTLY AND
LOOKING AT THE EDGE STRUCTURE
WITHIN THESE.
YOU'LL NOTE THAT IN -- WE'VE GOT
AN R -- VERY PRESENT AND
PROMINENT IN THE RAT AND PRETTY
MUCH ABSENT IN THE HUMAN, BUT WE
DO SEE -- CONNECTIVITY WITH A
VERY SIMILAR STRUCTURE IN THE
HUMAN DATA, SO IT STARTS ALLOW
YOU TO SAY MAYBE THESE ARE
FUNCTIONALLY ORTHOLOGUS AND
EVOLUTIONARILY THERE'S BEEN A
DIVERGENCE -- AND FINALLY, THIS
IS I THINK THE MOST EXCITING
THING ABOUT WHAT YOU CAN DO WITH
CONNECTIVITY DATA.
THIS IS A SORT OF A FRAMEWORK
WE'RE HOPING TO PUBLISH THIS
SOON BUT THE ACTUAL -- IF
ANYBODY IS INTERESTED IN DOING
THIS IT'S VERY EASY TO GENERATE
THIS ON YOUR DATA.
WE STARTED REALIZING YOU CAN
ACTUALLY USE CONNECTIVITY DATA
TO EXPLORE TISSUE MORPHOGEN --
YOU CAN SEE HERE HOW SPECIFIC
LIGAND TO RECEPTOR INTERACTIONS
WITH ARE VERY TIGHTLY ORGANIZED
IN THE TISSUE.
YOU CAN EVEN SEE ON THE BOTTOM
HERE THAT A SINGLE LIGAND, IN
THIS CASE, WINT5A BOTH LEFT AND
RIGHT, CAN BE SENSED VERY
DIFFERENTLY BASED ON SPECIFIC
REA ACCEPTOR PR ACCERECEPTOR PR.
I THINK THIS IS PROBABLY VERY
RELEVANT TO PULMONARY
REGENERATIVE MEDICINE, WE SEE A
STRUCTURAL REORGANIZATION OF THE
LUNG AND WE SEE THESE CELLULAR
CHANGES, AND I THINK THIS IS A
GREAT WAY TO LOOK AT AN
ADDITIONAL LAYER OF TISSUE
SECTIONS, BUT WE -- AND HOW
THOSE MIGHT CORRELATE.
SO THIS IS JUST MY CONCLUSIONS
HERE.
CELL-TO-CELL COMMUNICATION IS
REALLY OF HIGH IMPORTANCE TO
TISSUE BIOLOGY AND TISSUE
REGENERATIVE MEDICINE.
I THINK IT'S KIND OF
UNDERSTUDIED BUT IT DOESN'T HAVE
TO BE.
THERE'S AB A AN ENORMOUS AMOUNTF
INFORMATION THERE.
THERE'S A COUPLE OTHERS COMING
OUT NOW, WHERE YOU CAN REALLY
STATISTICALLY DO FLEXIBLE
ANALYSIS.
EVERYTHING I'VE SHOWN YOU, I'M
JUST CREATING A NEW DATA LAYER,
PUTTING IT BACK IN AND PLAYING
WITH IT.
THE SOFTWARE PAPER AND VIGNETTES
ARE HERE AND THEY'RE PUBLICLY
AVAILABLE, AND YOU CAN ALSO
EMAIL US IF YOU'RE INTERESTED IN
LEARNING HOW TO USE THESE.
AND I DON'T HAVE TIME SO WE CAN
TALK ABOUT THIS IN THE
DISCUSSION, BUT I THINK THERE
ARE MANY QUESTIONS THAT I
HAVEN'T ADDRESSED ABOUT
INTRACELLULAR TRANSDUCTION,
ABOUT CAUSAL INFERENCE IN THESE
DATA WHICH IS EXTREMELY TRICKY
AND I THINK WE NEED TO THINK
ABOUT IT WHEN WE -- AND THE
FINAL THING IS, THERE'S AN
OPPORTUNITY FOR SPAISH YOE
TEMPORAL MODELING AND REALLY
ADVANCED STATISTICAL METHODS AS
WELL AS -- THIS IS JUST MY
ACKNOWLEDGMENTS, I THANK MY LAB,
MY DEPARTMENT, FUNDING FOR DOING
THIS AND LOOKING FORWARD TO
TALKING AT THE END.
THANKS.
>> THANKS SO MUCH.
SO AS USUAL, YOU CAN PUT
QUESTIONS IN THE CHAT AND WE'LL
HAVE TIME FOR DISCUSSION AT THE
END.
AND SO NEXT WE HAVE SHAWN
DAVIDSON FROM NORTHWESTERN
UNIVERSITY, WHO WILL TALK ABOUT
SPECIAL METABOLOMICS.
>> THANKS TO THE ORGANIZERS.
I'VE REALLY EP JOYED THESE TWO
DAYS.
IT'S EXCITING TO SEE ALL THE
INTEREST IN METABOLISM.
SO TODAY I'LL TALK ON OUR WORK
IN USING SPATIAL METABOLOMICS
AND FLEX OMICS TO GET
INFORMATION AND GET DOWN TO KIND
OF THE SINGLE CELL LAYER.
SO WE KNOW THERE ARE MANY
FACTORS THAT INFLUENCE CELL
METABOLISM AND I COME AT THIS
FROM A CANCER METABOLISM
BACKGROUND, BUT WHEN WE THINK
ABOUT THIS, WE THINK ABOUT CELL
INTRINSIC FACTORS, SUCH AS
GENETIC ALTERATIONS IN THE
CANCER WORLDS, TUMOR GRADE OR
ONCOGENOTYPE, BUT ALSO THINGS
LIKE CELL LINEAGE AND TISSUE OF
ORIGIN, AND THESE THINGS ARE
SOMEWHAT EASIER, MAYBE NOT TUMOR
GRADE BUT TO STUDY IN CELL
CULTURE.
BUT THEN THERE'S ALSO THESE
EXTRINSIC FACTORS, SUCH AS
ACCESS TO NUTRIENTS AND OXYGEN
AND HOW THESE CELLS ARE
INTERACTING WITH OTHER CELLS AS
WE JUST HEARD IN THE LAST TALK.
A LOT OF MICROBIAL INTERACTION,
FROM MICROBIOME AND DIFFERENT
DIET INPUTS THAT WE HAVE.
SO WE NEEDED TO DEVELOP SOME
TOOLS TO STUDY METABOLISM BETTER
AND TO REALLY ACCOUNT FOR SOME
OF THESE EXTRINSIC FACTORS, SO
WE BORROWED SOME OF THIS FROM
THE DIABETES FIELD USING THE
HYPER -- UTILIZING A CLAMP AND
WE PUT CHRONIC CATHETERS INTO
ANIMAL MODELS OF DISEASE, INTO
THE JUGULAR VEIN AND INTO THE
CAROTID ARTERY WHERE WE CAN THEN
INFUSE STABILIZED TO LABELED
NUTRIENTS BE, AND THESE ARE NICE
BECAUSE THESE ARE SAFE AND
THERE'S ACTUALLY A NUMBER OF
PEOPLE INFUSING IN HUMAN
CLINICAL TRIALS AND WE CAN
SIMULTANEOUSLY SAMPLE THE BLOOD,
AND IT'S IMPORTANT TO DELIVER IT
THIS WAY SO YOU CAN GET SOME
PSEUDOISOTO PICK STEADY STATE
WHICH WILL MAKE THE MODELING
DOWNSTREAM OF THESE REACTION
NETWORKS EASIER.
JUST ONE STRONG DRAMATIC EXAMPLE
IS WHEN WE DID THIS IN A KRAS
P53 MOUSE MODEL OF CANCER, WE
USED CELLS AND WE INFUSED MICE
WITH THAT CANCER AND THEN
ISOLATED CELLS FROM THOSE SAME
ANIMALS.
WHEN WE LOOKED AT THE METABOLISM
IN VITRO, YOU SEE WHAT YOU WOULD
EXPECT, YOU SEE THIS STRONG
CANONICAL WARBURG EFFECT OF ALL
GLUCOSE CONVERTED TO LACTATE AND
YOU START TO USE GLUTE MEAN TO
FILL A TCA CYCLE.
BUT WHEN WE LOOKED AT THOSE SAME
TUMORS IN VIVO BY BULK
METABOLOMICS, YOU CAN SEE THE
TUMOR WILL STILL TAKE UP A LOT
OF GLUCOSE AS WE WOULD KNOW AND
EXPECT FROM FTG PET BUT THE FATE
OF THAT GLUCOSE IS DIFFERENT AND
SENDS MORE OF A -- NOT AS
RELEVANT IN THE PHYSIOLOGICAL
SENSE SO WE THOUGHT THIS MIGHT
BE A GOOD APPROACH.
SO NOW WE CAN START DOING THIS
ACROSS SCALES.
SO USING THAT INFORMATION,
STARTING AT THE LEFT, YOU CAN
GET WHOLE BODY METABOLISM SO YOU
CAN INFUSE STABLE ISOTOPES, YOU
CAN THINK ABOUT HOW NUTRIENTS
ARE CLEARED IN DIFFERENT
SETTINGS, THE CLEARANCE RATES.
ONCE YOU KNOW A NUTRIENT IS
BEING METABOLIZED DIFFERENTLY IN
A MOUSE MODEL, YOU CAN FOCUS ON
A TISSUE THROUGH BULK
METABOLOMICS.
AND TODAY WHAT I WANT TO FOCUS
ON IS OUR USE OF MULTI-IMAGING
MASS -- TO GET PRETTY CLOSE TO
SINGLE CELL LEVEL WITHIN
INDIVIDUAL TISSUES.
WE'VE STARTED TO WORK ON AND
THERE'S SEVERAL PUBLICATIONS OUT
ON USING EXPANSION MICROSCOPY TO
INCREASE THE RESOLUTION DOWN TO
SUBCELLULAR AND EVEN ORGANELLE.
SO THIS IS MORE OR LESS AN FAQ
SLIDE THAT COMES UP, SO WE RELY
FOR METABOLISM WORK ON FRESH
FROZEN TISSUES.
WE CAN USE FFP FOR PEPTIDES AND
LIPIDS.
WE HAVE ABOUT A 10 TO 50-MICRON
LATERAL RESOLUTION.
WE CAN MAKE RELATIVE OR ABSOLUTE
MEASUREMENTS, YOU KNOW, GET REAL
QUANTITATIVE NUMBERS FOR THE
CONCENTRATION OF A DRUG IN A
CERTAIN CELL POPULATION, AND WE
CAN START TO FOCUS ON SPECIFIC
CELL SUBSETS.
SO MALDI STANDS FOR MATRIX
ASSISTED LASER DESORPTION
IONIZATION MASS SPECTROMETRY
IMAGING.
WHICH MATRIX YOU SEE WILL DEPEND
ON WHICH METABOLITES YOU ARE
ABLE TO DETECT.
IT'S AN EXCITING AREA BECAUSE I
THINK EACH WEEK AND MONTH
THERE'S A NUMBER OF NEW MATRICES
THAT ALLOW US TO SEE MORE OF THE
METABOLISM THAT WE COULD SEE
BEFORE.
AND THERE'S DIFFERENT CHEMICAL
STRATEGIES INCLUDING -- AND WE
ROUTINELY WORK WITH NUMEROUS
MATRICES IN THE LAB TO GET THE
COVERAGE THAT MIGHT COME CLOSE
BUT NOT QUITE TO LCMS AT THIS
POINT.
IMPORTANTLY, WE'VE OPTIMIZED
THIS FT-ICR TECHNOLOGY.
IN IS A 9.4 TESLA MRMS SYSTEM
FROM BRUKE.
R AND IF YOU -- I THINK IF YOU
CLICK, SO YOU CAN SEE THIS IS
ACQUISITION IN ACTION, YOU GET A
NEW MASS SPECTRUM, AND THIS IS
SPECIFICALLY UNTARGETED SO WE
HAVE A LOT OF FEATURES WE HAVE
AL GATED AND ANNOTATED AND WE
HAVE A LOT OF OTHERS WHERE WE
DIDN'T NECESSARILY KNOW WHAT THE
MESS OF THIS COMPOUND IS, AND
CRITICAL FOR ALL OF THIS IS THAT
THE MASS RESOLUTION OF ALL THESE
INSTRUMENTS IS QUITE HIGHER THAN
THE TIME OF FLIGHT IMAGING
ASPECTS AND ALLOWS US TO -- SO
YOU CAN ACTUALLY LABEL A 13C AND
13N AND THEN -- THE DIFFERENCE
BETWEEN THOSE TWO ATOMS IN THE
SAME RUN.
SO THIS IS JUST ONE EXAMPLE.
IF YOU INFAWS A FULLY LABELED
GLUCOSE AND IT GOES THROUGH
GLYCOLYSIS, YOU WOULD GET A SIX
LABEL UDP GLUCOSE.
ON THE OTHER HAND, IF YOU
INHUGHES A THREE LABELED
GLYCEROL, YOU WOULD BE ABLE TO
MONITOR A THREE LABEL GDP
GLUCOSE AND THAT WOULD INDICATE
THAT GLUCOMUTAGENESIS IS
HAPPENING IN THAT REGION.
SO YOU CAN SEE HERE IN THE
MEDULLA, THERE'S MUCH MORE
GLYCOLYSIS HAPPENING.
THEN THE CORTEX, THERE IS MUCH
MORE GLUCOGENESIS HAPPENING.
SO WE CAN START TO INFER
DIFFERENT METABOLIC PATHWAY
ACTIVITIES.
SO THIS ISN'T LIMITED TO GLYCOL
GIST AND GLUCOGENESIS, YOU JUST
HAVE TO DESIGN THE TRACERS THAT
YOU'RE TRACING INTO TO LAND ON
THE METABOLITES THAT ARE
IMPORTANT FOR THE PATHWAY OF
INTEREST.
YOU A 'NOANOTHER EXAMPLE, AND IK
THIS HIGHLIGHTS WHAT WE DO
DIFFERENT THAN MANY OTHERS AT
THIS POINT, IF YOU LOOK AT THE
FAR RIGHT WHERE YOU HAVE MALE
UNLABELED, IF YOU LOOKED AT MA
LATE LEVELS, YOU WOULD SAY THESE
ARE ROUGHLY THE SAME IN BOTH
REGIONS SEPARATED BY THAT DARK
OUTER STRIPE.
BUT WHAT'S MORE INTERESTING IS
IF YOU ASK WHERE DOES THAT MA
LATE COME FROM, AND SO WEE
LOOKED AT GLUCOSE, AS DATE AND
WE LOOK AT THE -- EACH ONE OF
THOSE TO THE -- YOU CAN SEE
THERE'S NO REAL DIFFERENCE.
OARCHED, THERE'S SOME REBINARY
DIFFERENCES SO IF YOU LOOK AT
CITRATE, SO CITRATE CONTRIBUTES
TO MA LATE UNIQUELY IN THE
QUARTILE OF THE KIDNEY, THERE'S
ACTUALLY A -- IN THE CORTEX OF
KIDNEY THAT ISN'T IN THE
MEDULLA.
THE POINT IS THAT EVEN THOUGH
THE LEVELS LOOK THE SAME, THE
SOURCE OF THOSE NUTRIENTS ARE
DIFFERENT.
SO IF YOU'RE THINKING ABOUT
TARGETING METABOLIC PATHWAYS,
IT'S IMPORTANT TO KNOW IF YOU
WANT TO DRIVE THE LEVEL -- AND
WE CAN GET MORE QUANTITATIVE
ABOUT THAT AND PRETTY MUCH
ACCOUNT FOR ALMOST ALL OF THE
CARBON CONTRIBUTION TO MA LATE,
FOR EXAMPLE, IN THESE TWO
RENALES SO IF YOU LOOK AT THE
CORTEX, WE'RE CLOSE TO 100%
THERE AND WITH HIGHER SPATIAL
RESOLUTION THAN WE HAVE NOW,
WE'RE GETTING CLOSER TO NOT JUST
CORTEX AND MEDULLA BUT T-CELL, B
CELL, CANCER CELL, STROMAL CELL,
THINGS LIKE THAT, SO THIS IS
VERY IMPORTANT FOR GETTING
QUANTITATIVE NUTRIENT -- IN
DIFFERENT CELL COMPARTMENTS.
SO THOSE WERE CARBON LABELED
EXAMPLES AND JUST AS ONE OTHER
INTERESTING EXAMPLE, LOOKING AT
WHERE DOES THE NEUROTRANSMITTER
GLUTAMATE COME FROM IN THE
BRAIN.
AND SO WHERE DOES IT GET THE
NITROGEN TO MAKE GLUTAMATE?
AND THIS IS JUST STRIKINGLY
HETEROGENEOUS THROUGHOUT.
SO -- AND EVEN THOUGH THE LEVELS
OF GLUTAMATE ARE RELATIVELY
CONSISTENT ACROSS THESE BRAIN
REGIONS, HOW IT ACTUALLY GETS
THAT NITROGEN OUT OF IT IS QUITE
DIFFERENT.
AS ANOTHER EXAMPLE, THERE'S A
LOT OF EXCITEMENT IN DIFFERENT
WAYS TO EXPAND THE METABALOME
COVERAGE, AND THIS WAS A SERIES
OF EFFORTS ON IMPROVING
DETECTION USING A WASHING
STRATEGY TO REMOVE THINGS THAT
MIGHT CAUSE ION EXPRESSION, SO
FOR THE FIRST TIME USING A
WASHING STRATEGY, WE'RE AGE TO
DETECT QUANTITATIVELY BY
SMAICIAL METABOLOMICS THE LEVEL
OF NAD AND NADH AND THIS ALSO
ENHANCES THE SIGNAL IN GENERAL
OF OTHER PHOSPHOTATE CONTAINING
METABOLITES, AND STRATEGIES LIKE
THIS CAN ACTUALLY BE USED TO SEE
A NUMBER OF DIFFERENT MOLECULES
AND THINKING IT'S AN EXCITING
AREA TO BE FOCUSING ON.
SO JUST TO THINK ABOUT SOME
CURRENT GAPS IN WHAT WE'RE DOING
HERE, WHEN WE HAVE STARTED TO
LOOK AT SOME PATIENT SAMPLES, I
THINK IT'S GOING TO BE IMPORTANT
TO FIND SOME WAYS TO DO BATCH
CORRECTION THAT ACCOUNT FOR BOTH
THE WAY THE SAMPLES ARE
COLLECTED AND THE TECHNICAL
VARIABILITY THAT HAPPENS RUN TO
RUN AND MONTH TO MONTH ON MASS
SPECS.
THIS IS NOT AN UNFAMILIAR
PROBLEM TO THE MASS SPEC WORLD,
SO WE'RE WORKING ON THIS AND
TRYING TO MAKE THIS WORK FOR
OUR -- AS WELL.
WE CONTINUE TO -- THIS IS
SOMEWHAT LABORIOUS BECAUSE WE
DON'T HAVE THE TEAM TOGRAPHY ON
THIS IMAGING PLATFORM YOU WOULD
HAVE WITH LCS SO WE RELY A LOT
ON MS2 AND ON TISSUE MS2, AND I
THINK THERE'S A LOT OF ROOM FOR
IMPROVEMENT IN KIND OF
TISSUE-SPECIFIC METABOLIC MODELS
AND WE'RE WORKING ON THAT
THROUGH A SERIES OF EXPERIMENTS
USING KINETIC TRACING IN THIS.
WITHIN MY LAB, WE'RE ESPECIALLY
INTERESTED IN UNDERSTANDING LUNG
CANCER HETEROGENEITY.
WE'VE STARTED TO DO SOME
EXPERIMENTS WITH CELL AUTONOMOUS
AND NON-CELL AUTONOMOUS RESPONSE
TO INFECTION IN COLLABORATION
WITH INVESTIGATORS HERE, AND
WE'RE GENERALLY JUST GETTING THE
DATA TO TALK TO EACH OTHER,
DOING SOME OF THESE ASSAYS ON
THE SAME SLIDE AND GETTING THE
CELL TYPE-SPECIFIC METABOLISM OF
DIFFERENT CELLS IN THE LUNG IN
DIFFERENT CONTEXTS.
SO I THINK THE FUTURE IS
BRIGHTER, I WAS INSPIRED BY A
LOT OF TALKS HERE AND SO I LOOK
TO DISCUSS FURTHER.
AND TO THANK THE LAB WHICH IS
JUST GETTING STARTED, AND IF
ANYONE IS INTERESTED MASS SPEC,
PLEASE FEEL FREE TO EMAIL ME.
I'D BE HAPPY TO CHAT MORE.
THANKS SO MUCH.
>> THANK YOU FOR THAT TALK.
LACKING FORWARD TO DISCUSSION IN
A MOMENT.
WANTED TO GO AHEAD HERE AND
INTRODUCE OUR LAST SPEAKER FOR
THIS SESSION, PROFESSOR JIANWEN
QUE FROM COLUMBIA UNIVERSITY
MEDICAL CENTER.
>> YOU PRONOUNCED THE CHINESE
VERY WELL, THANK YOU.
I'D ALSO LIKE TO THANK THE
ORGANIZING COMMITTEE FOR THIS
WONDERFUL WORKSHOP, AND I HAVE
BEEN LEARNING A LOT FOR THE PAST
TWO DAYS.
AND TODAY I WOULD LIKE TO TALK
ABOUT THIS POSSIBLE ROLE OF THE
NEURON IN LUNG REGENERATION, AND
HERE I USE POSSIBLE BECAUSE IT
REALLY IS.
NOT MANY SOLID DATA OUT THERE TO
SHOW THAT THERE'S A ROLE PLAYED
BY NEURON IN LUNG REGENERATION.
SO IN THIS SHORT TALK, I WOULD
LIKE TO DIVIDE MY TALK INTO
THREE PARTS.
FIRST I'M GOING TO TALK ABOUT
THIS INNOVATION OF THE TRACHEA
AND THE LUNG AND THEN TALK ABOUT
THIS POTENTIAL ASSOCIATION OF
INNERVATION WITH LUNG
REGENERATION BUT BASED -- MOSTLY
BASED ON DATA FROM OTHER ORGANS,
GUT AND INTESTINE, AND BUILD ON
THOSE FINDINGS IN THEIR TISSUES
AND I WAS THINKING ABOUT WHAT
POSSIBLE GAPS AND CHALLENGE THAT
WE WERE FACING WITH OUR TISSUE
AND LUNG.
SO THE -- SENSORY NEURONS
INCLUDE THIS VAGUS NEVER,
WHICH -- NEURON BODY LOCATED IN
THE BRAINSTEM SPECIFICALLY --
AND TUBULAR GANGLION, SEND
FIBERS T TO INNERVATE THE AIRWAS
AND THE LUNGS.
AND THE OTHER SIDE OF THE CENTER
OF THE NEURONS COMING FROM THIS
DORSAL ROOT GANGLION, THEY CAN
ALSO -- AIRWAYS.
THE MOTOR NEURONS ARE SENT BY
THIS SUR CAL THORACIC -- AN THEY
SEND THESE MOTOR NEURONS AND
NERVE FIBERS INNOVATE MUSCLES
AND BLOOD VESSELS IN THE LUNG.
THIS IS A VERY BEAUTIFUL PICTURE
TO SHOWN -- AT OREGON UNIVERSITY
HEALTH SCIENCE CENTER, HE USED
THIS PGP ANTIBODY, 9.5 ANTIBODY
TO LABEL NEURONS, NERVE FIBERS
IN THE LUNG AND YOU CAN SEE THIS
INTRICATE NERVE FIBERS
DISTRIBUTE ALONG THE TRACHEA BUT
ALSO THE AIRWAYS AND ALSO -- AND
RUN THROUGH THESE ALF VEE LIES.
AND THIS IS CREATING LABELED
TOMATO -- VIE GUS NERVE FIBERS
IN THVAGUS NERVE FIBERS INTHE L-
SOME OF THESE FIBERS SPREAD INTO
THE ALVEOLAR REGION, AND THE
PICTURE IN THE MIDDLE IS NA YOUN
SEE SOME OF THIS -- L STANDS FOR
LUMEN AND YOU CAN SEE THE
EPITHELIUM, LABELED WITH THIS E.
SHOWN BY THIS UNDERLYING NERVE
GUY PERT THIS IS ANOTHER -- THAT
STEPHN TO SHOW THAT THESE
LABELED NEURONS ARE LOCATED
RUNNING AROUND THE AL A V ALVEOD
AIRWAYS SHOWN BY THIS BRIGHT
STAINING WITH THIS GFP.
WOUWE ALL KNOW THIS IS ONE OF TE
OBVIOUS CELL WHICH IS NEURON --
THEY ARE INNERVATED ALSO BY
VAGUS NERVE AS WELL HERE, SHOWS
THIS NEURON SUBTYPE LABELED BY
P2RY1.
YOU CAN SEE THIS STAINING TO
SHOW THESE NERVE FIBERS
INNERVATE TO THE NEW
NEUROANTIBODY NEUROEPITHELIUM
BODIES.
THIS ARE ALSO INNERVATED BY
OTHER TYPES OF NEURONS, FOR
EXAMPLE, -- ALSO LABELED VAGUS
NERVE, AND YOU CAN SEE THAT THEY
ARE UNDERLYING THIS AND ANOTHER
PROTEIN CALLED CALCIUM BINDING
PROTEIN, YOU CAN SEE THAT THE
VAGUS NERVE CAN RUN THROUGH
THIS.
THOSE VAGUS NERVES CAN ALSO
INNERVATE ALL -- WHICH IS
GENERAL MARKER FOR VAGUS NERVE.
AND SENSE REA NERVE.
SENSORY NERVE.
WE DON'T KNOW WHETHER THEY
CONTACT WITH TYPE 2 CELLS BUT
YOU CAN SEE THEY'RE VERY CLOSE
TO TYPE 2 CELLS AND TYPE
1 CELLS, AND SHOWN BY THE TYPE E
STAINING AND THEN TYPE 1 CELLS
SHOWN BY THIS GREEN STAINING.
AND THERE'S ANOTHER -- SHOULD BE
CRE MOS1 -- INNOVATE ALVEOLAR
REGION, ALTHOUGH WE DON'T KNOW
WHETHER IT CAN FORM SYNAPSE WITH
THESE EPITHELIAL CELLS OR WHAT
KIND OF THINGS ARE BEING
INNERVATED, IT JUST REMAINS
UNKNOWN.
SO WE KNOW THESE NERVES PLAY AN
IMPORTANT PART IN THE LUNGS AND
TRACHEA AND IT CAN REGULATE
BREATHING, IT CAN ALSO REGULATE
MUCUS SECRETION AND BRONCHOMOTOR
TONES AND ALSO REFLEX, ALSO
COUGH -- SO BOTH CAN CONTROL
BLOOD VESSELS TONES AND UPON
STIMULATION.
AND THEN -- HAS SHOWN VAGUS
NERVE, ALONG WITH OTHERS, HAS
SHOWN VAGUS NERVE CAN MODULATE
THESE IMMUNE FUNCTIONS, AIRWAY
HYPERACTIVITIES AS WELL, SHE HAS
A VERY NICE PAPER TO SHOW
CIRCULATE FROM THE LUNG TO THE
BRAIN TO MEDIATE THIS IMMUNE
FUNCTIONS AND TYPE OF ACTIVITY
FUNCTIONS OF THE LUNG.
AND ALONG WITH OTHERS THEY'VE
SHOWN THIS HAS BEEN ASSOCIATED
WITH THIS CHRONIC DISEASE, FOR
EXAMPLE, HYPERREACTIVITIES AND
MUCUS OVERPROTECTION AS WELL,
BECAUSE THERE IS A CLOSE
INTERACTION BETWEEN GOBLET CELLS
AND NEURON FIBERS.
MY LAB IS STARTING TO GET INTO
THIS AREA MORE ON THE CANCER
SIDE AND IN PART IS MOTIVATED BY
MY COLLEAGUE, TIMOTHY WONG, WHO
HAS IDENTIFIED THAT IMPORTANT
ROLE OF THE VAGUS NERVE IN STO
MA'AM CANCER, AND WHAT HE FOUND
IS IF YOU CUT THE VAGUS NERVE,
YOU ACTUALLY COULD INHIBIT TUMOR
GROWTH IN THE STO MA'AM.
HE ALSO HAD OTHER PAPER AND SHOW
THAT VAGUS NERVE ALSO PLAYS AN
IMPORTANT ROLE IN -- CANCER.
SO WHAT WE FOUND IS THAT DURING
SMALL CELL LUNG CARCINOMA
DEVELOPMENT, AND YOU CAN SEE
GRADUALLY ACCUMULATION OF THIS
SYMPATHETIC NEURON NERVE FIBERS
IN DIFFERENT STAGE OF TUMOR
FORMATION, AND IN THE MIDDLE
PANEL, YOU CAN SEE THAT --
ACCUMULATED NEURON FIBERS,
NERVE -- IN A CLOSE RANGE OF
THOSE IN SITU LUNG CARCINOMA AND
WITH DISEASE PROGRESSION YOU CAN
SEE THOSE FIBERS NOW INFILTRATED
AND MIGRATE INTO THE TUMOR
TISSUE, WHICH IS SHOWN ON THE
RIGHT SIDE.
SO WE ASK CAN WE BLOCK THIS
TUMOR GROWTH BY JUST GET RID OF
THESE NEURONS.
HERE WE USE THIS 6-OHDA TO
ABLATE THOSE NERVE FIBERS IN
MOUSE TISSUE WITH THOSE
INJECTION -- WITH FOUR DOSES
INJECTION THEY CAN SEE NOW THIS
NERVE FIBER HAS BEEN ABLATED
WHICH IS SHOWN ON THE RIGHT SIDE
IN IN SITU TUMOR.
AND WHEN WE WAIT FOR A LONG
TIME, YOU CAN SEE THOSE TUMOR
NOW IS DISAPPEARED OR SHRINKED,
OHDA AND YOU CAN QUANTITATE THEM
AND YOU CAN SHOW THAT THERE'S
REDUCTION IN TUMOR SIZE ALSO IN
HUMOR NUMBERS IN A MOUSE TREATED
WITH 60.
6-OHDA SUGGESTING THAT
THERE'S A NEUROAND CANCER CELL
INTERACTIONS DURING TUMOR
GROWTH.
BUT UNFORTUNATELY, REALLY RIGHT
NOW WE DON'T HAVE TOO MUCH DATA
ABOUT FOR THIS -- SO WE HAVE TO
LOOK INTO OTHER TISSUES.
IN THE INTESTINE, IT HAS BEEN
SHOWN BY FREDERICK AND
GENENTECH, HE SHOWS THAT
ADRENERGIC NERVES CAN PRODUCE
NEURAL TRANSMITTERS AND GROWTH
FACTORS WHICH IN TURN CAN
STIMULATE -- AND ALSO STEM
CELLS, THE IMMUNE CELLS, HERE HE
SHOWS THAT -- JUST
NEUROTRANSMITTERS OR GROWTH
FACTORS STIMULATE BY -- SE CITED
BY THE NEURONS CAN WORK ON TYPE
3 INNATE LYMPHOCYTE, WHICH IN
TURN CAN PRODUCE THIS
INTERLEUKIN 22, INTERLEUKIN 22
AND NOW CAN WORK ON THOSE STEM
CELLS TO -- EXPANSION AND ALSO
DIFFERENTIATE INVOLVED IN THIS
REGENERATION.
ANOTHER GROUP IN BEJING SHOWS
THAT ANOTHER TYPE OF NEURONS
WHICH IS 5HT,
5 HYDROXYTRYPTAMINE ALSO CAN
PROMOTE TUMOR GROWTH VIA THE
INTERACTION WITH THE CANCER STEM
CELLS, AND WHAT THEY SHOW IS
THAT MICROBIOTA CAN STIMULATE
NEURON -- NERVE ENDINGS TO
PRODUCE THIS 5HT.
SEROTONIN, TRANSMITTER WHICH IN
TURN CAN WORK ON -- STEM CELLS
AND DRIVE THE ACTIVATION OF --
WHICH IN TURN FURTHER PROMOTE
THE EXPANSION OF CANCER TEM
CELLS AND PROMOTE TUMOR
FORMATION.
NNT SAME GIN THE SAME GROUP THAE
PROLIFERATION VIA THE
MICROPHAGES IN THE MIDDLE THEY
USE THIS HE ENTERIC NEURONS THAT
AGAIN THOSE M -- CAN ACTD ARE
ATIONACTIVATENEURONS WHICH CAN T
WHICH IN TURN CAN PRODUCE
MICROPHAGE AND PGE2 LATER ON CAN
WORK ON THE STEM CELLS TO DRIVE
PROLIFERATION.
MANY OF YOU IN THE AUDIENCE IS
SHOWN THERE ARE MULTIPLE STEM
CELLS IN THE LUNG INCLUDING
BASAL CELLS, EPITHELIAL CELLS
AND ALSO WE HAVE SHOWN --
NEGATIVE POPULATIONS CAN DRIVE
LUNG REGENERATION.
BUT WE HAVE NO IDEA HOW THESE
NEURONS INTERACT WITH THESE STEM
CELLS, AND WHETHER IT IS
FLUID -- OR -- WHICH HAVE
BEHAVED LIKE A NEURON NETWORK
COMPONENT AND ALSO -- AND ALSO
IN ALVEOLI, WE DON'T KNOW
WHETHER -- WE DON'T KNOW WHETHER
THOSE NERVE ENDINGS CAN DIRECTLY
AFFECT THE ENDOTHELIAL CELLS OR
OTHER CELL TYPES AND WE DON'T
KNOW WHETHER THEY ARE DIRECT OR
INDIRECT THROUGH THESE -- IMMUNE
CELLS.
SO THERE ARE MANY GAPS, AND --
GAPS, AS I SAID WE DON'T KNOW
WHETHER NEURONS PLAY ANY ROLE IN
REGENERATION AND LACK OF
UNDERSTANDING THE NEURONS
SPECIFIC INNOVATING THESE
AIRWAYS AND WE DON'T KNOW
WHETHER THOSE -- WHAT TYPE OF
INCREASED NEURONS APPEARED IN
THE -- IN ASTHMA AND WE HAVE NO
IDEA HOW EXACTLY THIS
NEUROENDOCRINE CELLS INVOLVED IN
EPITHELIAL REGENERATION AT A
MOLECULAR LEVELS, AND THIS IS
LARGELY UNKNOWN AND ALSO LACK OF
IMAGING SYSTEM TO SHOW THE NERVE
ENDINGS IN EPITHELIAL CELLS.
SO WE HAVE MANY QUESTIONS WE
NEED TO ADDRESS AND CAN DISCUSS
LATER ON IN THE DISCUSSION PART.
WITH THAT, I WOULD LIKE TO THANK
MY PEOPLE IN THIS PICTURE
ESPECIALLY -- WHO GAVE ME LOTS
OF INPUT AND PREPARED THESE
SLIDES.
THANK YOU.
>> THANK YOU VERY MUCH.
SO NOW WE'D LIKE TO TRANSITION
INTO THE DISCUSSION OF GAPS AN
OPPORTUNITIES PARTICULARLY
FOCUSED ON Q & A SESSION WITH
THE SPEAKERS THAT WE'VE HAD IN
THIS LAST SESSION.
JUST A HOUSEKEEPING NOTE, I KNOW
WE'RE A LITTLE BIT OVER TIME.
WE WILL HAVE A BRIEF BREAK JUST
TO HELP US TRANSITION AT 2:45 TO
OUR -- THAT'S 2:45 EASTERN TO
OUR STRATEGIC DISCUSSION, WHERE
WE'LL WRAP UP AND HAVE A FEW
FORWARD LOOKING POINT ON THE
ENTIRE WORKSHOP.
SO I'D LIKE TO OPEN THE FLOOR UP
TO ANY QUESTIONS FOR OUR
SPEAKERS IN THIS PARTICULAR
PANEL AT THIS POINT.
>> SOMEONE RAISED THEIR HAND.
PLEASE.
>> HI.
THANKS FOR THE GREAT TALKS.
ALL.
PERHAPS MORE DIRECTED TOWARDS
MICHA BUT EVERYONE CAN RESPOND.
I'M CURIOUS HOW YOU THINK ABOUT
THE FUTURE OF COMPUTATIONALLY
MODELING COMPLEX DISEASES LIKE
THE VARIOUS FORMS OF LUNG
DISEASE YOU MENTIONED WHEN IT
FEELS LIKE EVERY DAY, WE END UP
DISCOVERING A NEW CELL STATE OR
TRANSITIONAL STATE, I MEAN, FROM
MY ENGINEERING BACKGROUND, YOU
CAN'T BUILD A MODEL IF THE
NUMBER OF VARIABLES JUST KEEPS
INCREASING AND CHANGING AND YET
THAT SEEMS TO BE THE SITUATION
WE'RE IN, SO HOW DO YOU SQUARE
THAT WITH EVERYONE'S DESIRE TO
BUILD MORE PREDICTIVE MODELS OR
STATISTICAL MODELS OF WHAT'S
GOING ON IN TISSUES?
>> I REALLY LIKE THIS QUESTION.
I THINK IT'S A GREAT QUESTION,
HE'S SPOT-ON, IT'S AN ACTIVE
CHALLENGE AND PROBABLY ONE OF
THE FOREMOST CHALLENGES RIGHT
NOW BUT IT IS PROBABLY
SURMOUNTABLE.
THE KEY THING IS TO RELLIZE
THERE ARE THESE CATEGORIES OF
SCALE YOU CAN GO UP IN.
20, 30 YEARS AGO PEOPLE MIGHT
HAVE SAID OH IT'S TOO MUCH TO BE
STUDYING MULTIPLE MOLECULES
WITHIN A SINGLE CELL.
NOW WE'VE GOT OMICS LEVEL THINGS
WHERE THE PATTERNS THAT WE ARE
STUDYING AND WE'RE USED TO
STUDYING WHEN WE'RE LOOKING AT
MAPS OF CELL STATE AND YOU KNOW
EACH IS A VECTOR OF TENS OF
THOUSANDS OF NUMBERS AND WE'RE
SEEING HOW THOSE SELL STATES
CHANGE AND ARE REDISTRIBUTED.
WELL, THERE'S ANED AD LEVEL THAT
WE CAN GO UP WHEN WE START
TALKING ABOUT THINGS LIKE TISSUE
STATE, WE LOOK AT THE ACTUAL --
SO LIKE HOW MANY OF EVERY CELL
TYPE THERE IS WITHIN A TISSUE,
WITHIN A REGION, WE LOOK AT THE
INDIVIDUAL CELL STATE
THEMSELVES, WHAT'S THE
REPRESENTATION, AND THEN WE ALSO
LOOK AT THE CONNECTIVITY VALUES,
HOW THEY COMMUNICATE.
ALL OF THAT CAN BE CONDENSED OR
COMPRESSED DOWN INTO EMBEDDINGS
IN THE SAME WAY THAT WE NOW LOOK
AT CELL STATE, WE CAN START TO
LOOK AT THINGS LIKE TISSUE
STATE.
AND WHEN YOU HAVE THOSE KIND OF
A COMPUTATION -- THAT IS
COMPUTATIONAL TASK BUT ALSO SORT
OF A CONCEPTUAL ONE WHERE YOU
CAN SAY THIS IS CONTROL TISSUE,
IPF, COPD, REGENERATING TISSUE,
IF WE CAN START TO MAKE RELIABLE
EMBEDDINGS OF THE DATA, THEN WE
REALLY HAVE THE TOOLS NOT ONLY
TO DO PREDICTIVE MODELING BUT
ALSO JUST TO SORT OF OBSERVE OUR
DATA VERY CLEARLY AT THE END YOU
CAN SAY WHAT FEATURES ARE
DRIVING THAT, OH IT'S THE
EMERGENCE -- OR THE LOSS OF
THE -- IN THE LUNG OR THESE NEW
FIBROBLASTS.
BUT FIRST YOU CAN JUST SORT OF
SEE HOW THESE STATE TRANSITIONS
ARE OCCURRING AND HOW WE MIGHT
BE ABLE TO TARGET THEM
THERAPEUTICALLY.
SO THERE'S THIS BRIDGE THAT HAS
TO HAPPEN BETWEEN FEATURE LEVEL
INVESTIGATIONS AND MUCH MORE
HOLISTIC SYSTEMS LEVEL
COMPRESSION AND EMBEDDING AND
ARCHITECTURES THAT ALLOW US TO
OBSERVE OUR DATA.
WE'RE GETTING THERE, THOUGH.
THIS IS GOING TO HAPPEN SOON, I
THINK.
I DON'T KNOW IF OTHER PEOPLE
HAVE COMMENTS ABOUT THIS TOO.
IT'S A VERY GENERAL SUBJECT.
>> I THINK A RELATED QUESTION,
SO WITH A LOT OF THE -- THIS IS
ALL VERY TRANSCRIPTOMIC BASED,
WHERE DO YOU THINK -- WHEN DO
YOU THINK OR WHERE ARE WE MOVING
TO ACTUALLY DO SIMILAR STUDIES
BUT AT THE PROTEOMIC LEVEL?
IS THAT LIKE A PIPE DREAM OR IS
THAT MORE OF A POSSIBILITY IN
THE COMING YEARS?
>> I THINK IT'S VERY POSSIBLE.
I THINK THE TECHNOLOGIES ARE
IMPROVING.
YOU CAN ALREADY GET SOME REALLY
BEAUTIFUL MULTIPLEX SPATIAL
PROTEOMICS FROM LIKE CODEX
PLATFORMS AND OTHER THINGS LIKE
THAT.
YOU CAN USE EXISTING ANTIBODIES,
THERE'S A LOT OF DIFFERENT WAYS
OF DOING THIS.
I THINK IT HARDER -- JOSE MIGHT
KNOW MORE ABOUT IT, IT'S NOT AS
WELL DEVELOPED BUT IT IS
BEAUTIFUL DATA AND I THINK IT'S
VERY INTERESTING TO LOOK AT THE
TRANSCRIPTOMIC LAYER AND THE
PROTEOMICS LAYER BECAUSE THEY
ARE NOT ALWAYS THE SAME, AND
THEY'RE NOT THE SAME IN SOME
VERY INTERESTING WAYS.
WHEN YOU LOOK AT RNA, YOU'RE
LOOKING AT PROTEIN PRODUCTION --
IT LIKE A PROXY FOR FLUX, HOW
MUCH IS BEING SORT OF -- HOW
MUCH IS IT PATHWAY ACTIVATED?
THAT'S WHERE YOU GET THE RNA
LEVEL.
ON A PROTEIN LEVEL, IT MAY BE
BECAUSE YOU'RE DEGRADING THE
PROTEIN AND YOU'RE NOT SEEING IT
ALL.
SO IT'S VERY PATHWAY-SPECIFIC,
VERY PROCESS SPECIFIC.
I THINK WE DO NEED THESE
MULTIPLE LAYERS OF INFORMATION
TO REALLY UNDERSTAND WHAT'S
GOING ON.
>> MAYBE JUST TO KIND OF SHIFT
THE CONVERSATION OVER TO SHAWN,
YOU KNOW, WE WERE JUST TALKING
ABOUT RNA AND PROTEIN METABOLISM
PROBABLY FITS IN THERE
SOMEWHERE.
HOW DO YOU FORESEE SOME OF THE
METABOLIC FLUX THINGS THAT
YOU'RE DOING IN TERMS OF
INFLUENCING OUR UNDERSTANDING OF
THESE POTENTIAL RECEPTOR LIGAND
INTERACTIONS REGARDLESS OF WHAT
WE'RE IB FERRING THEM FROM?
>> I THINK THERE'S A LOT TO
LEARN HERE.
I THINK THE STRENGTH OF WHAT
WHICH DO IS THAT, YOU KNOW,
GETTING METABOLIC ACTIVITY
WILL -- THERE SHOULD BE A PRETTY
DECENT DISCONNECT FROM WHAT YOU
SEE FOR PROTEIN OR RNA LEVELS,
JUST BECAUSE OF DIFFERENT
ENZYMATIC ACTIVITIES THROUGH
THIS, AND SO I THINK IT ADDS A
VERY COMPLEMENTARY LAYER OF
INFORMATION AND HOW DIFFERENT IT
WILL BE FROM WHAT YOU SEE AT THE
PROTEOMIC LEVEL OR WHAT IT WILL
PREDICT OUTSIDE OF THAT, I'M NOT
SURE, BUT -- AND WE'RE REALLY
EXCITED TO INTEGRATE THESE
THINGS.
>> ABSOLUTELY.
I THINK ONE QUESTION THAT CAME
UP ACTUALLY BEFORE -- WE CAN
MOVE AHEAD TO YOHANNES, PLEASE.
>> THANK YOU.
THOSE WERE GREAT PRESENTATIONS.
MY QUESTION IS FOR SHAWN.
SHAWN, WHEN YOU LOOK AT SPECIAL
METABOLOMICS ON, SAY, INFLAMED
TISSUE, NOT HEAVILY INFLAMED BUT
SOME INFLAMMATION, WOULD YOU
REALLY SEE KIND OF COLOR CHANGES
OR REALLY CHANGES THAT ONE CAN
QUANTIFY, OR IS IT SOMETHING
THAT YOU NEED TO HAVE, LIKE A
DRASTIC RESPONSE TO SEE THESE
KIND OF METABOLOMICS CHANGES?
>> I DON'T THINK IT HAS TO BE SO
DRASTIC.
I THINK IT DEPENDS ON WHAT THE
CAUSE OF INFLAMMATION IS.
WE HAVE SOME EARLY DATA ON
INFECTION MODELS AND SO WE HAVE
SOME LOCAL INFORMATION AROUND
THERE.
WE'VE DONE A LOT OF TUMOR WORK
AS WELL, SO I DON'T THINK THEY
HAVE TO BE DRAMA TI AND I THINK
THE OTHER STRENGTH OF
METABOLOMICS IS THAT THESE
CHANGES HAPPEN VERY QUICKLY.
WE'VE BEEN USING THIS
IMPLANTABLE MICRO DEVICE IN
CLINICAL TRIALS, AND THE DEVICE
IS IN THE TUMOR FOR ABOUT THREE
HOURS, AND WE CAN ACTUALLY GET
MEANININGFUL CELL RESPONSES IN
JUST THAT AMOUNT OF TIME, IT
MIGHT TAKE LONGER TO SEE
TRANSCRIPTIONAL CHANGES.
SO I THINK BOTH THE SPEED AND
THE CHANGE FROM THOSE DIFFERENT
INFLAMED AREAS COULD BE HELPFUL.
AND POSSIBLE.
>> VERY INTERESTING.
SO IN OTHER WORDS, LET'S SAY,
FOR EXAMPLE, IF YOU HAD A TIME
COURSE, AS YOU MENTIONED, LET'S
SAY, YOU KNOW -- EXPOSED SOMEHOW
SOMETHING AND LACKS OVER TIME.
AND ARE THERE DATA, THERE'S SOME
KIND OF CORRELATION DATA, SAY,
FOR EXAMPLE, IN TERMS OF LAVAGE
TO IT AND THIS KIND OF
METABOLOMICS OR SOMETHING LAKE
THAT, THOSE KIND OF DATA THAT
ARE GENERATED ALREADY OR NOT?
>> THAT MAY EXIST AT THE BULK
LEVEL.
WE ARE ASKING SIMILAR QUESTIONS
ON THE SPATIAL LEVEL.
>> OKAY.
THANK YOU.
>> QUESTION FOR JIANWEN.
HAVE YOU STARTED LOOKING INTO
EITHER THE SEROTONIN OR OTHER
TRANSMITTER RECEPTORS -- CELLS?
>> THAT'S A GOOD QUESTION.
WE ARE JUST BEGINNING TO LOOK
INTO THIS, AND CREATE MODELS AND
THE FIRST THING WE'RE LOOKING AT
SINGLE CELL, I'M SURE YOU MAY
HAVE ACCESSED ALL THIS DATA AS
WELL, TO LOOK NE SINGLE CELL
DATA AND SEE WHAT ARE THE
POTENTIAL NEUROTRANSMITTERS
EXPRESSED OR RECEPTORS EXPRESSED
IN EPITHELIUM CELLS AND THE
MESENCHYMAL CELLS.
THE SEROTONIN, WE HAVE NOT
PARTICULARLY PAID ATTENTION TO.
WHY DO YOU ASK THE QUESTION?
WHY SEROTONIN IS IMPORTANT DO
YOU THINK?
>> THERE'S A FEW REPORTS
SUGGESTING SOME OF THE SEROTONIN
SUBTYPES -- RECEPTOR SUBTYPES
EXPRESSING TYPE 2 CELLS, AND --
SOME SHOWING MORE, SOME SHOWING
LESS.
>> I SEE.
INTERESTING.
>> I HAVE A QUESTION.
CAN I ASK A QUESTION?
>> OF COURSE.
>> SO THIS IS FOR JIANWEN AS
WELL, I HAVE A QUESTION ABOUT
THE NEURON WORK.
DO YOU THINK THIS CAN BE USED --
THAT -- I MEAN, YOU'VE GOT
BEAUTIFUL PICTURES THERE, AND I
DON'T KNOW MUCH ABOUT THIS AND
I'M JUST SORT OF SEEING THIS
INCREDIBLE NEURON NETWORK IN THE
LUNG, AND IT BRINGS TO MIND, I
FORGET -- I'D HAVE TO DIG
THROUGH LITERATURE BUT I KNOW
THERE'S SOME STUDIES WHERE CAN
YOU ESSENTIALLY DOSE NEURONS INA
VERY VO WITHINVIVO AND THEY'LL T
DELIVERED AT THE SYNAPSES.
CAN YOU USE THIS AS A DELIVERY
MECHANISM TO TRIGGER
REGENERATION IN THESE TISSUES OR
TO MANIPULATE THE CELLS WHEREVER
THE END PROCESSES ARE?
>> THAT IS A VERY GOOD QUESTION.
I WOULD JUST SAY, THOSE
BEAUTIFUL PICTURES ARE COMING
FROM OTHER LABS AND FROM --
OTHER LABS AS WELL.
MY LAB, WE JUST -- THE PAPER WAS
JUST ACCEPTED YESTERDAY I THINK
BY CANCER DISCOVERY AND WE DO
SEE THAT NERVE ENDINGS
INTERACTING WITH CANCER CELL,
BUT TO COME BACK TO YOUR
QUESTION, YES, YOU CAN USE THAT
TOOL TO DELIVER DRUGS THROUGH
THE NERVE FIBERS AND TO THE --
FROM GANGLIA INTO THE BRAIN.
AND THOSE COULD BE USE THE AND A
VIRUS AND AS A CARRIER TO DRIVE
ALL THOSE AGENTS.
THAT'S INTERESTING.
>> I HAD A QUESTION FOR HELEN.
I WAS WONDERING, WE HEARD A
LITTLE BIT, I'M BY NO MEANS AN
EXPERT MYSELF IN THE INTEGRATED
STRESS RESPONSE, BUT WE HEARD
YESTERDAY IN ONE OF THE FIRST
TALKS ABOUT THE IMPORTANCE OF
THE INTEGRATED STRESS RESPONSE
KIND OF DOWNSTREAM OF
MANIPULATING THE ELECTRON
TRANSPORT CHAIN IN MITOCHONDRIA
AND THEN LEADING TO PARTICULAR
SUSCEPTIBILITY IN TERMS OF LOSS
OF AL VE ALVEOLAR EPITHELIAL CE.
I DON'T KNOW IF YOU WERE IN
ATTENDANCE FOR THIS TALK OR IF
YOU HAD ANY REFLECTIONS ON YOUR
WORK BASED ON THIS ISR MECHANISM
AND SIMILARITIES OR DIFFERENCES
BETWEEN LUNG PARENCHYMA AND WHAT
YOU'RE SEEING IN PERIPHERAL
NEURONS.
>> YEAH, I MEAN, IT'S
INTERESTING, BECAUSE I THINK
PEOPLE ARE JUST STARTING TO --
PEOPLE HAVE STUDIED IT FOR MANY
YEARS IN CANCER, CANCER-LIKE
CELLS, BUT I THINK IMPLICATIONS
IN OTHER DISEASES AS WELL AS
DEVELOPMENT AS IT WAS EMPHASIZED
YESTERDAY, WE'RE JUST STARTING
TO WORK ON.
I THINK ONE OF THE POINTS
THAT -- I THINK IT WAS DR. HAN
THAT GAVE THAT TALK THAT SHE
ALSO RAISED THAT WE ALSO DON'T
KNOW THE SEPARATION BETWEEN --
WE KNOW ONCE YOU ACTIVATE STRESS
RESPONSE YOU HAVE THESE
SIMULTANEOUSLY ACTIVATION OF
SPECIFIC TARGETED GENES, AS WELL
AS YOU HAVE A SHUTDOWN IN GLOBAL
TRANSLATION SO YOU HAVE A
DEPLETED PROTEIN SYNTHESIS.
AND WE DON'T KNOW WHICH ONE IS
ACTUALLY -- WE DON'T EVEN KNOW
WHICH ONE IS RESPONSIBLE FOR THE
PHENOTYPES WE'RE SEEING AND I
THINK SHE ALLUDED TO THAT, AND
WE DON'T KNOW WHICH ONE OF THOSE
ARMS IS ACTUALLY RESPONSIBLE FOR
THAT EITHER.
SO I MEAN, I DON'T HAVE A
STRAIGHT ANSWER FOR THAT.
YOU KNOW, JUST TO SAY THAT WE
STILL HAVE A LOT TO UNDERSTAND
AND STUDY.
THERE IS ONE OF THE FOCUS OF OUR
LAB, NOW I TRY TO SEPARATE THESE
ARMS AND TRY TO UNDERSTAND HOW
EACH ONE OF THEM AFFECT IN OUR
CASE THE --
>> AND I GUESS RELATED MORE
GLOBALLY TO THE USE OF IN VITRO
MODELS FOR STUDYING ALS
PATHOGENESIS, I WAS WONDERING IF
YOU COULD COMMENT A LITTLE BIT
IN TERMS OF YOUR PERSPECTIVE ON
HOW THE NEUROSCIENCE COMMUNITY
HAS KIND OF BEEN PUSHING FORWARD
ON THE DEVELOPMENT OF THESE
MODELS.
WE HEARD A VERY NICE TALK IN THE
PRIOR SESSION THIS MORNING ABOUT
THE USE OF MODELS TO STUDY KIND
OF MONOGENIC CASES OF DISEASE.
ARE WE STILL LIMITED TO
MONOGENIC CASES?
DO WE SEE -- SHINING THROUGH IN
THESE CULTURES, WHAT'S YOUR
FEELING ON THIS?
>> I MEAN, WE'RE MOSTLY USING
MONOGENIC, JUST BECAUSE WE HAVE
AT LEAST 38 DIFFERENT GENES
ASSOCIATED TO ALS THAT WE KNOW
ARE CAUSATIVE.
BUT WE HAVE MOVED SORT OF TO TRY
AND INCREASE THE NUMBER OF
SPORADIC PATIENTS, AND SOMETIMES
I DO GET ASKED WHEN I GIVE THIS
TALK, LIKE ARE YOU INCLUDING
SPORADIC PATIENTS.
IN ORDER TO INCLUDE THOSE CELLS,
WE NEED A LARGER NUMBER.
SO ONE PERSON THAT HAS BEEN
WORKING A LOT ON THAT IS CLIVE
SWENSON AT CEDARS-SINAI, SO HE
DOES HAVE A GOOD REPOS TRIF, SO
YOU JUST HAVE TO INCREED BUT
WHAT WE SAW IS THAT A LOT OF
THESE PHENOTYPES THAT WE SEE
MODELS, WE CAN OFTEN
RECAPITULATE AT LEAST PARTIALLY,
SO ANOTHER LINE OF STUDY SINCE
WE'VE IDENTIFIED THE I - --
BECAUSE OF TIME, SO THESE TWO
ACTIVATORS ARE CURRENTLY IN
CLINICAL TRIAL FOR SPORADIC AND
FOR FAMILIAL ALS CASES.
THEY'RE ALSO IN CLINICAL TRIAL
FOR VANISHING WHITE MATTER
DISEASE.
SO EVEN IF YOU START FROM
DIFFERENT MONOGENIC OR EVEN IF
YOU START FROM DIFFERENT
MOLECULAR EVENTS YOU CAN END UP
WITH THE SAME SORT OF PATHWAY IF
YOU ACTIVATE ISR.
SO WE ARE TRYING NOW TO IDENTIFY
WHICH ONES ARE ACTUALLY BEING --
WHICH OF THESE GENES CAN BE
ASSOCIATED TO THIS ACTIVATION OF
ISR.
WE HAVE STUDIED FIVE OF THEM AND
HALF OF THEM DOES -- DIFFERENT
MUTATIONS, DIFFERENT VARIANTS.
SO I THINK THAT'S HOW WE ARE
PROGRESSING.
ANOTHER THING THAT WE'RE DOING
IS A LOT OF PEOPLE HERE ALSO
MENTIONED THE USE OF ORGANOIDS
AND OTHER SYSTEMS, SO IN OUR LAB
WE DO A CO-CULTURE SYSTEM,
BETWEEN THE MOTOR NEURONS AND
SKELETAL MUSCLE.
WE HAVE BEEN ABLE TO
SPONTANEOUSLY GENERATE THESE
NEUROMUSCULAR JUNCTIONS, THE
SYNAPSES BETWEEN THE TWO, AND I
THINK THAT'S WHAT WE'RE
PROGRESSING TOWARDS, SO NOT JUST
USING THESE SILOED SPECIFIC CELL
TYPES, BUT ALSO INCORPORATING
OTHER CELLS TO THE SYSTEM.
>> IN TERMS OF INCORPORATING
OTHER CELLS, I KNOW, AMANDA, NOT
ONE OF THE WEEKERS, BUT HERE IN
THIS SESSION, I WANTED TO KIND
OF PULL YOU INTO THE DISCUSSION
HERE.
WE'VE HEARD A LITTLE BIT ABOUT
IMMUNE CELLS IN A FEW OF THE
DIFFERENT TALKS BUT ALWAYS
PLAYING QUITE A PERIPHERAL ROLE
HERE.
I KNOW YOUR LAB IS THINKING A
LOT ABOUT IMMUNE CELL
CONTRIBUTIONS AND HELPING TO --
I WAS JUST WONDERING, YOUR
GENERAL VIEW IN TERMS OF HOW YOU
SEE THE IMMUNE SYSTEM, IN TERMS
OF BEING ABLE TO DRIVE TISSUE
RESILIENCE?
>> THAT'S SOMETHING I WAS KIND
OF THINKING ABOUT THROUGHOUT THE
PAST COUPLE DAYS, AND SOME
CONNECTIONS HERE AND THERE BUT
TALLLY THAT WAS A GOOD QUESTION
I HAD FOR -- MAYBE SPECIFICALLY
JIANWEN.
SO IN TERMS OF WHAT THE NEURONS
ARE DOING IN CANCER, DO YOU KNOW
SPECIFICALLY WHAT CELLS THEY'RE
INTERACTING WITH AND WOULD THAT
HAVE -- ARE THEY INTERACTING
WITH IMMUNE CELLS?
I THINK THAT WAS ONE OF THE OPEN
QUESTIONS, BUT CAN YOU SORT OF
SPECULATE ON WHAT YOU'RE SEEING
IN CANCER AND HOW THAT COULD
INTERACT OR HOW THAT COULD
REFLECT ON WHAT COULD BE
HAPPENING IN MORE PROGRESSIVE
LUNG DISEASES THAT -- AND WHAT
CELLS ARE REALLY INTERACTING
WITH AND WHAT THEIR CELLS
THEY'RE CONTROLLING.
THAT'S A VERY OPEN-ENDED
QUESTION, I THINK, BUT THIS IS
THE TIME TO SPECULATE, HOW WOULD
YOU GO ABOUT TRYING TO FIGURE
THAT OUT AS WELL.
-- FOR ALL THE SPEAKERS, HOW
DOES IT INTERACT WITH THE IMMUNE
SYSTEM.
I GUESS ALSO IN TEMPLES BEING
ABLE TO -- YOU SEE THESE
SNAPSHOTS IN TIME, THE IMMUNE
CELLS ARE COMING IN AND OUT, SO
I THINK THAT'S ANOTHER
COMPLICATION IN THESE SYSTEMS
WHEN YOU'RE LOOKING AT THINGS.
>> THAT'S A VERY GOOD QUESTION,
ONE CHALLENGE I WAS THINKING I
WANT TO PUT ON IS HOW WOULD THAT
REGENERATION -- IMMUNE CELLS, WE
KNOW THEY ARE COMING VERY
QUICKLY.
VIRUS INFECTIONS, THE NEURONS
ARE ACUTELY INJURED.
THEY ARE NOT QUIESCENT, THEY
QUICKLY COME OUT OF THIS
QUIESCENT STATE AND FIRE SIGNALS
AND IN THE FIRST INTERACT
MESENCHYMAL CELLS AROUND THEM
AND BLOOD VESSELS WHICH CAN
CONTROL THE BLOOD VESSELS AND
ALSO MAY POTENTIALLY INTERACT
WITH PATROLLING MICROPHAGES OR
TYPE 2 INNATE LYMPHOID CELLS AS
WELL, AND RIGHT NOW I HAVE NO
EVIDENCE BUT AS I JUST MENTIONED
IN THE INTESTINE, THOSE NERVE
FIBERS CAN INTERACT WITH
MICROPHAGES BUT ALSO INTERACT
WITH THE ROC2 CELLS.
WE RECENTLY IDENTIFIED THIS IN
ANOTHER POPULATION,
RIC3 POPULATION, SPECIFICALLY
LOCALIZED TO THE BIG BLOOD
VESSELS.
WE DON'T KNOW WHETHER THEY ARE
INVOLVED IN CONNECTION WITH
THESE NERVE FIBERS AND MY
COLLABORATOR HAS A VERY
INTERESTING DATA TO THOUGH
THESE, AND ALSO SURVEY IMMUNE
RESPONSE ENVIRONMENT TO --
>> I THINK ONE OF MY FAVORITE
EXAMPLES OF THIS, I REMEMBER
NOW, HEARING QUITE A LONG TIME
AGO FROM SOME WORK THAT RICH
LOCKSEED'S GROUP WAS DOING IN
TERMS OF THE POSITIONING OF
ILC2s AT THESE CRITICAL BRANCH
IF WAVES, POSITIONING -- NEURONS
AS WELL KIND OF CONTROLLING THIS
OVERALL COMMUNITY.
THE HYPOTHESIS YOU COULD ALMOST
ENVISION CLOSING OFF CERTAIN
REGIONS OF THE LUNG IN CASE THAT
CERTAIN DANGER WAS DETECTED TO
SHIFT FLOW FROM ONE DIRECTION OR
ANOTHER.
I DON'T KNOW IF THIS STILL
REMAINS PURELY SPECULATIVE, BUT
SINCE WE TALKED ABOUT
REGIONALITY, I WAS WONDERING IF
THE PANELISTS HERE HAVE HAD ANY
THOUGHTS ON THIS IF
>> I NOTICED YOU HAD SOME IMMUNE
CELLS -- NEURONS IN YOUR
DATASET?
>> I DO NOT IN MY DATA.
I WISH I DID.
IF ANYONE HERE HAS DATA FROM THE
LUNG THAT HAS A CLEAR NEURON
POPULATION IN IT, I THINK IT
WOULD BE VERY INTERESTING TO
LOOK AT THIS, AND I STRONGLY
SUSPECT THEY HAVE A UNIQUE
LIGAND AND RECEPTOR PRO T PROFID
WE COULD DOPE IN
NEUROTRANSMITTERS SO YOU COULD
SEE WHO'S LISTENING OR CAPABLE
OF LISTENING.
THE DATA I HAVE DOES NOT HAVE
NEURONS IN IT AND I -- BUT
DEFINITELY WITH THE RIGHT DATA
YOU COULD LOOK AT THIS.
>> ONE QUESTION I HAD IS RELATED
TO THE TIME COMPONENT.
THIS MAY COME UP MORE OF AN
EPIDEMIOLOGICAL LEVEL OVER THE
THEME OVER THE PAST FEW
SESSIONS.
BUT IN TERMS OF GENERATING TIME
SERIES DATA THAT IS AMENABLE FOR
THE TYPES OF COMPUTATIONAL
METHODS THAT YOU'VE DEVELOPED,
HOW REFINED DO WE NEED TO BE?
BECAUSE I FEEL LIKE IF WE DO A
TIME SEARS EASE SOMETIME SERIES,
JUST BECAUSE OF REALITIES, OF
FUNDING, THREE OR FOUR TIME
POINTS IN BETWEEN.
THEN YOU WERE LIKE MAN, HEY, I
THINK IF I'D PROFILED EIGHT
HOURS I WOULD HAVE KNOWN EVEN
MORE.
HOW MUCH TIME RESOLUTION DO WE
NEED?
>> THE FUNDING -- PEOPLE WANT TO
SEE TIME POINTS, THEY DON'T WANT
TO SEE REPLICATES.
WE MAKE THOSE DATASET AND THAT'S
SORT OF THE STANDARD HERE AND
YOU CAN SEE VERY STRONG
SIGNALING, BUT AS I THINK YOU'RE
POINTING OUT, IT IS MUCH, MUCH
MORE BEAUTIFUL AND FUN TO PLAY
WITH THE DATA IF YOU -- LIKE I
WOULD RATHER HAVE FEWER
REPLICATES AND A TIGHTER TIME
COURSE, BECAUSE EACH INDIVIDUAL
TIME POINT HAS ENORMOUS BLEED A
IN PHENOTYPE SPACE.
IT'S LIKE A FRACTION OF THE
SHELLS HAVE -- SO THAT ONE TIME
POINT YOU GET AT DAY 3 ACTUALLY
HAS SOME CELLS THAT ARE VERY
SIMILAR TO WHAT YOU SEE AT DAY
ONE AND DAY 7 MAYBE.
SO YOU GET A DISTRIBUTION.
SO THAT MEANS YOUR STATISTICAL
POWER IS MUCH, MUCH STRONGER.
IF WE WERE DOING TIGHT TIME
SERIES OF THESE KINDS OF
PROCESSES, THE DATA QUALITY IS
BETTER.
BUT YOU END UP HAVING TO WRITE
THESE GRANTS THAT ARE SAYING I'M
GOING TO DEVOTE ALL MY RESOURCES
TO DOING A SINGLE TIME POINT OR
N OF 2 TIGHT TIME COURSE, SO YOU
END UP REDISTRIBUTING AND SAY
I'M GOING TO DO AN N OF 6, SO I
HAVE A STRONG ANSWER TO THAT
ABOUT WHAT WE SHOULD BE DOING
SCIENTIFICALLY, BUT IT CAN BE
CHALLENGING TO -- THIS IS
EXPENSIVE DATA TO MAKE.
TIME COURSE DATA IS BEAUTIFUL.
>> SO IT LOOKS LIKE WE'RE
GETTING SOME QUESTIONS COMING
THROUGH THE CHAT, SO MAYBE LAST
10 MINUTES, WE CAN START GOING
THROUGH THIS.
SAM, HAVE YOU COMPARED NICHES
WITH CELL PHONE DB OR CELL CHAT
D ABOUT ARE THESE TOOLS FOCUSED
ON NODES OR EDGES?
>> ANYONE WHO'S GOING TO START
DOING CONNECTIVITY ANALYSIS, YOU
LITERALLY HAVE LIKE 40 OR 50
DIFFERENT TOOLS TO PULL FROM.
THOSE BEING MENTIONED ARE SOME
OF THE EARLIEST ONES AND THEY'RE
SORT OF THE GOLD STANDARD,
THEY'RE EASY TO PULL OFF THE
SHELF.
WE'VE DONE THIS COMPARISON
EXTENSIVELY.
THE BASIC ANSWER IS THAT ALL OF
THESE DIFFERENT TOOLS ARE GOING
TO GIVE YOU KIND OF SORT OF
SIMILAR ANSWERS BUT THEY'RE
DIFFERENT IN TERMS OF HOW EASY
OR HARD THEY ARE TO USE AND HOW
STATISTICALLY ADAPTABLE THEY
ARE.
AND THAT'S THE BIG DIFFERENCE.
IF YOU USE THE SAME GROUND
TRUTH, YOU QUERY THE SAME
MECHANISMS, THESE TOOLS ARE ALL
GOING TO GIVE YOU MORE OR LESS A
PRETTY SIMILAR PORTRAIT OF
WHAT'S GOING ON.
BUT IF YOU -- NICHES, THERE'S A
NEW ONE CAME OUT CALLED DEEP
TOK, THOSE ARE ALL GOING TO GIVE
YOU SINGLE EDGE DAY, I FIND OUT
WOW, IT'S NOT FROM OUR TIME
COURSE BUT FROM OUR DRUG EFFECT
OR SOMETHING.
YOU CAN REALLY QUICKLY PIVOT AND
CHASE THAT PATTERN IN THE DATA.
WITH THE NODE BASED ONES LIKE
CELL CHAT AND CELL PHONE DB THAT
HAVE BEEN MENTIONED HERE, YOU
PUT HANDCUFFS ON AT THE BEG
BEGINNING -- REALLY THE
DIFFERENCE COMES WITH HOW
STATISTICALLY FLEXIBLE THEY ARE
AND HOW EASY THE DATA ANALYSIS
IS.
BUT AT THE END OF THE DAY, IF
YOU DO ALL THESE THINGS RIGHT
YOU CAN GET THESE TOOLS TO AGREE
WITH ONE ANOTHER.
>> DR. MIRANDA, CAN YOU PLEASE
COMMENT ON WHAT THE FIELD THINKS
ABOUT HOW RSRIB IMPROVES -- I
WONDER WHAT IS THE PROPOSED
MODEL IN THE NEUROFIELD?
>> THAT'S INTERESTING.
THAT'S A GREAT QUESTION.
THE SHORT ANSWER IS THAT I DON'T
HAVE A PROPOSED MECHANISM.
I ENDED UP HAVING A MEETING WITH
THE PERS PERSON LEADING THE CLIL
TRIALS, AND THE DATA WAS
COLLECTED BASED ON THESE MOUSE
MODELS, I THINK IT SHOULD BE
KNOCKOUT MOUSE MODELS, KIND
OF -- THE MECHANISM OF ALS, BUT
THE ONLY DATA FROM AN ALS MODEL
WHICH IS FROM THE SOD1, THIS IS
THE -- I GUESS THE BEST MODEL
THAT WE HAVE SO FAR, THE BEST
MOUSE MODEL THAT WE HAVE, DID
NOT ENCOURAGE THE USE OF ISRID.
SO WHEN I HAD THAT CONVERSATION
HER INITIALLY, SHE SAID THEIR
INVETO DATA WERE SHOWING VITRO
JUST LIKE OURS DID, AND THEY
WERE THINKING ABOUT PROPOSING A
MISLOCALIZATION OF THIS PROTEIN
THAT GOT PHOSPHORYLATED AND THIS
LOCALIZES AND IT'S COMMON TO
MOST OF THE ALS, IT'S A PA
WILLING TO THAT IS SEEN MOST
WITH -- BUT IN OUR MODEL
SPECIFICALLY, THE FACT IT
DOESN'T HAVE THAT PATHOLOGY.
SO IN THIS CONVERSATION WITH
HER, SHE'S LIKE OH MAYBE WE
SHOULD LINK THAT TO A
MITOCHONDRIA DYSREGULATION.
SO I THINK -- OUR WORK IS THE
FIRST ONE THAT SHOWS A SPECIFIC
VARIANT THAT IS LINKED TOLL
PHENOTYPE THROUGH ACTIVATION OF
ISL.
SO THE FIELD DOESN'T HAVE THAT
YET.
>> SO IT MORE OF LIKE THE NEURAL
FIELD MORE OF THE ISSUE OF --
ACTUALLY IMPROVED THE FUNCTION
OF WHATEVER THE CALL FOR CELL
TYPE.
WHAT WE SHOWED IS THE CHANGE FOR
DIFFERENTIATION POTENTIAL BUT
IT'S NOT NECESSARILY FUNCTION UR
ABOUT FOR IMPROVED FUNCTION OF
THE CELL.
THANK YOU.
>> THERE'S ONE LAST QUESTION
BEFORE WE WRAP UP.
OR MAYBE WE'LL HAVE TIME FOR A
COUPLE MORE BUT JUST A COUPLE
MORE MINUTES HERE AND ACTUALLY I
HAD THIS QUESTION AS WELL, SO
THIS IS FOR -- DO YOU OBSERVE
ANY COLOCALIZATION OF NEURONS
AND BLOOD VESSELS IN THE LUNG?
DO YOU THINK NEURONS MIGHT PLAY
A ROLE IN REPAIRING BLOOD
VESSELS AFTER INJURY?
>> YES.
BLOOD VESSELS CONTROLLED MAJORLY
BY SYMPATHETIC NEURONS AND IF
YOU STAIN WITH TH, YOU CAN SEE
THERE'S SMOOTH MUSCLE CELLS IN
THE BLOOD VESSELS, AND ALSO I --
ALSO SEND NERVE ENDING.
TOLS, THESE TWO -- CONSTRICTION,
THINGS LIKE THAT.
>> THANKS.
WELLINGTON CARDOSA HAS A
QUESTION.
RAISED HAND.
>> YES.
THIS QUESTION WAS FORMICA.
FOFOR MICHA.YOUR IDEA OF ACTUAL
EXPLORING -- IT'S NOT
CONTROVERSIAL BUT IT IS SUCH A
COMPLEX THING BECAUSE WHEN WE
HAVE A TISSUE MORPHOGEN IT
DEPENDS ON SO MANY DIFFERENT
THINGS.
THERE IS -- BETWEEN
EXTRACELLULAR MATRIX.
THERE ARE CORE RECEPTORS, AND I
WONDER WHAT IT IS INSIDE THIS
PAMMAGE --
>> THIS IS JUST GETTING STARTED.
SO I WOULD BE HAPPY TO TALK WITH
YOU ABOUT THIS MORE BECAUSE
YOU'RE PUTTING YOUR FINGER RIGHT
ON THE ISSUES.
WHAT IS VEABL NOW AVAILABLE NOE
PACK SJ IS VERY SIMPLE AND
YOU'RE RIGHT, CAN I CALL THAT A
TISSUE MORPHOGEN?
NO, I CAN CALL THAT A SIGNALING
FIELD OR COLOCATION OF -- AND
YOU CAN SEPA TESHES.
THAT'S REALLY WHAT WE CAN STUDY
CURRENTLY.
I WILL SAY I'VE GOT
CONVERSATIONS GOING ON WITH
STATISTICIANS WHICH ARE TRYING
TO ANSWER WHAT YOU'RE BRINGING
UP, WHICH IS FOR EACH -- WE NEED
TO THINK ABOUT INTERACTIONS WITH
OTHER MECHANISMS, AND WE HAVE TO
THINK VERY CAREFULLY ABOUT WHAT
WE CAN PULL OUT OF IT.
BUT THE INFORMATION IS THERE.
IT'S A MATTER OF THEN MAKING THE
RIGHT STATISTICAL MODELS THAT
ARE REALLY BIOLOGICALLY
INFORMED.
SO I THINK THERE'S ENORMOUS
POTENTIAL THERE IN THE COMING
DECADES TO DO THIS TO LIBRARIES
OF TISSUES IN PARAFFIN RIGHT
NOW, BUT WE DO HAVE TO DO IT
CAREFULLY.
>> I'D LOVE TO LEARN MORE ABOUT
THAT.
THANK YOU.
>> SO I THINK WHAT WE'RE GOING
TO DO IS I WON'T CALL IT A BREAK
BUT JUST SO PEOPLE CAN STAND UP
AND SIT BACK DOWN IN 2 1/2 TO 3
MINUTES.
WHY DON'T WE ALL GO AHEAD AND DO
THIS AND THEN WE'LL START AGAIN
SHARP AT 2:45 WITH THE STRATEGIC
DISCUSSION.
SO THANK YOU VERY MUCH TO THE
PANELISTS AND EVERYWITHIN FOR
EVERYONE FOR YOURQUESTIONS.
SO WHAT I WANTED TO DO IS
JUST SHOW A COUPLE OF SLIDES TO
FRAME THE DISCUSSION AND WRAP-UP
HERE.
SO HERE ARE SOME OF THE OVERALL
GOALS FOR THE WORKSHOP THAT WE
HAD AS WE WERE PUTTING THIS
TOGETHER.
FIRST AND FOREMOST TO REALLY
HELP FACILITATE COLLABORATIONS
AMONGST BASIC SCIENCE
RESEARCHES, CLINICIANS, INDUSTRY
EXPERTS AND REACHING SOME OF
THESE KNOWLEDGE GAPS IN OUR
UNDERSTANDING OF REGENERATIVE
STEM CELL RESILIENCE WITH
CHRONIC LUNG DISEASES.
WE ALSO NEED TO CONNECT RESEARCH
ON BASIC MECHANISMS TO WHAT
THERAPEUTIC NEEDS ARE ES PISHLY
AS THEY RELATE TO LUNG
PROGENITORS, RESILIENCE AND
CHRONIC LUNG DISEASE AND LASTLY
TRY TO PRIORITIZE WHAT THE
CRITICAL RESEARCH GAPS,
CHALLENGES AND OPPORTUNITIES ARE
AS THEY PERTAIN TO THESE TOPICS.
SO THE OVERALL WORKSHOP
OBJECTIVES PUT INTO A LITTLE
MORE GRANULARITY ARE TO HELP
DEFINE PROGENITOR CELL
RESILIENCE AND SOME OF THEIR
REGULATORY FACTORS TO UNDERSTAND
THE ROLE OF F OF PROGENITOR CELN
EARLY ONSET OF CHRONIC DISEASE,
DISCUSS EMERGING TECHNOLOGIES
AND MODEL SYSTEMS, IDENTIFY GAPS
IN RESEARCH ON PROGENITOR BASED
MARKERS PREDIBTING EARLY
DISEASE, DISCUSS THERAPEUTIC
INTERVENTIONS TARGETING
PROGENITORS TO PREVENT AND TREAT
THEM, LEVERAGE INSIGHTS FROM
OTHER DISCIPLINES AND AGAIN TO
FACILITATE COLLABORATION AMONG
BASIC SCIENCE CLINICIANS AND
INDUSTRY EXPERTS.
WHAT I THINK WE'VE BEEN ABLE TO
DO IS CERTAINLY OVER THE PAST
FEW YEARS, ENABLED BY BOTH A LOT
OF HYPOTHESIS-DRIVEN WORK AND
ALSO NEW AND EMERGING UNBIASED
PROFILING TECHNIQUES, WE'VE
BECOME QUITE GOOD AND DEFINING
PROGENITOR CELL STATES AND
SUBSETS, AND I THINK WE'VE HEARD
A LOT ABOUT THIS.
I THINK THAT THERE'S STILL THE
CHALLENGE OF CONNECTING THESE
PROGENITOR CELLS WITH RESILIENT
FUNCTIONS, AND SOME OF THIS MAY
BE DUE TO THE NEEDS OF BETTER
DEFINED RESILIENCE AND IN DOING
SO, HELP TO UNDERSTAND THE
REGULATORY FACTORS THAT HELP TO
PUSH AND PULL ON THE SYSTEM.
LIKEWISE, ONE OF THE TASKS, ONE
OF THE MANDATES THAT WE HAD WAS
TO TRY TO UNDERSTAND THE ROLE OF
PROGENITOR CELLS AND THE EARLY
ONSET OF CHRONIC DISEASE.
I THINK THIS IS WHERE A LOT OF
US ARE THINKING IN TERMS OF
BETTER PREVENTION AND EARLY
INTERVENTION AND INTERCEPTION OF
DISEASE.
HOWEVER, BY NECESSITY OF THE
TYPES OF DISEASE MODELS OR HUMAN
SYSTEMS THAT WE WORK IN, WE'VE
GAINED A LOT OF UNDERSTANDING OF
SOME OF THESE FEATURES IN REALLY
EXTREME CHRONIC DISEASE SO
TRYING TO WALK THAT BACK
EARLIER, GOING BACK TO THE VERY
BEGINNING IN RAVI'S VERY
BEAUTIFUL KEYNOTE TALK I THINK
IS SOMETHING OF UTMOST
IMPORTANCE.
THE NEXT THING WE'VE BEEN ABLE
TO DO IS WE CERTAINLY HAVE NOW
MUCH MORE ADVANCED MODEL SYSTEMS
THAN WE DID 10 OR 20 YEARS AGO,
BUT I THINK THERE'S STILL GAPS
IN A LOT OF THESE MODEL SYSTEMS,
AND THIS IS HIGHLIGHTED BY SOME
OF THE TALKS AND THEY MIGHT BE
GOOD FOR PERHAPS EARLY FETAL OR
KIND OF NEONATAL TISSUE
UNDERSTANDING, BUT REALLY THAT
ROBUST ADULT UNDERSTANDING IS
CHALLENGING AND OF COURSE
MULTISYSTEM MODELS CAN BE QUITE
COMPLEX TO DEVELOP.
WHILE WE HEARD SOME I THINK VERY
FASCINATING TALKS IN TERMS OF
BIOMARKERS CONNECTING THEM TO
PROGENITOR BASED BIOMARKERS IS
STILL A CHALLENGE, ESPECIALLY AS
THESE PROGENITOR CELLS ARE BEING
ACTIVELY DEFINED.
WE'VE HEARD ABOUT DEFINITIONS
THAT OFTENTIMES ARE IN RNA SPACE
OR PERHAPS VALIDATED THROUGH A
COUPLE OF KEY MARKERS THAT COULD
BE STAINED FOR OR USED FOR
LINEAGE TRACING, BUT HOW THAT
TIES TO PERIPHERAL CIRCULATING
BIOMARCS AND IF THOSE, IN FACT,
ARE COMING FROM THE PROGENITORS
OR LINKED TO THEIR FUNCTION, I
THINK IT'S STILL VERY MUCH AN
OPEN QUESTION.
THE NECK THING THAT WE HAVE OF F
COURSE THERAPEUTIC
INTERVENTIONS, WE OF COURSE NEED
MUCH MORE WORK DONE HERE.
IN OUR LAST SESSION, WE'RE
CERTAINLY ABLE TO CAPTURE SOME
INTERESTING THOUGHTS FROM OTHER
DISCIPLINES, BUT THERE'S MUCH
MORE TERRITORY TO BE EXPLORED
HERE.
AND WHILE WE HAD A VERY NICE
REPRESENTATION OF PANELISTS FROM
BASIC SCIENTISTS AND CLINICIAN
FIELD, I REALLY WANT TO MAKE
SURE IF THERE'S ANY BIOTECH OR
INDUSTRY REPRESENTATIVES IN THE
AUDIENCE TO REALLY HEAR YOUR
VOICE IN TERMS OF HOW WE CAN BE
BETTER FACILITATING
COLLABORATION.
SO RESILIENCE.
A WORD THAT WAS RIGHT IN THE
TITLE OF THIS WORKSHOP ON
PROGENITOR RESILIENCE IN THE
EARLY ONSET OF CHRONIC LUNG
DISEASES.
AND THE MORE I THINK ABOUT IT, I
THINK IT'S ACTUALLY REALLY A
PERFECT WORD TO CAPTURE THE
CHALLENGE OF WHAT WE NEED TO DO
HERE.
THE REASON FOR THIS IS BECAUSE
THERE ARE TWO DEFINITIONS FOR
THIS WORD.
THE FIRST ONE IS THIS CAPACITY
TO WITHSTAND OR RECOVER QUICKLY
FROM DIFFICULTIES AND A CERTAIN
TOUGHNESS, THAT PERHAPS THE
IMAGE ON THE LEFT OF THIS REALLY
BEAUTIFUL ART FORM OF POTTERY
WHERE CERAMIC IS SOMETHING THAT
IS QUITE DURABLE AND HARD UNTIL
IT BREAKS AND ONE KIND OF
PATCHES IT UP OR PUTS IT BACK
TOGETHER.
WE'VE HEARD A LOT OF TALKS ABOUT
TYPES OF PROGENITOR CELLS THAT
MAY ACT IN THIS WAY, TO HELP
PROVIDE THE LUNG WITH A CERTAIN
TOUGHNESS OR RAPID RESPONSE
MECHANISM, BUT THEN THESE
MECHANISMS COULD BE COOPTED IN
CHRONIC DISEASE.
THE OTHER ONE, AND I THINK THIS
IS SOMETHING THAT WE HEARD LESS
ABOUT, BUT SOMETHING THAT WE
REALLY NEED TO THINK CAREFULLY
ABOUT IS THIS ABILITY OF AN
OBJECT TO SPRING BACK INTO SHAPE
OR ELASTICITY.
THIS OF COURSE IS FUNDAMENTAL
WITH EVERY SINGLE BREATH THAT
WE'RE TAKING.
SO I THINK NOT JUST THINKING
ABOUT RESILIENCE AS KIND OF A
TOUGHNESS OF THE SYSTEM, BUT
ALSO THE PARTICULAR SHAPE OR
ELASTICITY IT CAN KEEP IS
SOMETHING THAT WILL BE VERY
IMPORTANT AS WE FRAME OUR
DISCUSSION AND THE CHALLENGES
GOING FORWARD.
SO WHAT I'D LIKE TO DO HERE IS
POSE A FEW QUESTIONS JUST TO
KIND OF OPEN UP DISCUSSION AND
WE CAN CERTAINLY BOUNCE BACK AND
FORTH BETWEEN THESE, AND I HOPE
AS MANY OF THE PANELISTS THAT
ARE ABLE TO STAY ON ARE ABLE TO
PARTICIPATE IN THIS STRATEGIC
DISCUSSION SECTION.
SO AS SOME THINGS COME UP,
PLEASE FEEL FREE TO CHIME IN.
WE'LL TRY TO USE THE RAISE HAND
FUNCTION JUST TO KEEP THINGS IN
GOOD ORDER.
BUT I THINK A FEW QUESTIONS AT
LEAST CAME TO MIND AS I WAS
LISTENING AND TRYING TO
INTEGRATE ALL THESE TALKS ARE,
IS RESILIENCE JUST THE ABSENCE
OF DISEASE OR IS IT REALLY THE
PRESENCE OF A SPECIAL STATE?
SO HOW DO WE BEGIN TO QUANTIFY
SOMETHING THAT WE MAY NOT EVEN
KNOW HOW TO MEASURE.
THIS IS SOMETHING THAT'S REALLY,
REALLY IMPORTANT TO TRY TO
UNDERSTAND.
I THINK MOST PEOPLE MAY THINK OF
STEM AND PROGENITOR CELL AS A
GOOD THING, BUT WE'VE ALSO HEARD
ABOUT SOME PLACES WHERE THERE
MIGHT BE INDIVIDUAL STATES WHICH
COULD BE POTENTIALLY
DETRIMENTAL, AND FURTHERMORE, WE
NEED TO UNDERSTAND THE
REGIONALITY OF WHERE THOSE CELLS
ARE LOCALIZED WITHIN THE LUNG.
THE NEXT QUESTION THAT COMES TO
MIND IS HOW WE BALANCE THE
COMPLEXITY AND THE PHYSIOLOGICAL
RELEVANCE WITH THE SCALABILITY
OF OUR SYSTEMS, AND SO ONE COULD
THINK ABOUT THIS IN TERMS OF
SOME OF THE EX VIVO MODELS OR IN
VIVO REDUCTIONIST MODELS THAT WE
USE TO STUDY HUMAN DISEASE BUT
ALSO THE TYPES OF MEASUREMENTS
THAT WE'RE TAKING IN HUMAN
TRIALS AS WELL.
THE NEXT QUESTION THAT COMES TO
MIND IS HOW WE CONNECT EARLY
KNOWLEDGE ON PROGENITOR
HETEROGENEITY TO NON-LUNG
PARENCHYMAL BIOMARKERS TO REALLY
ACCESS THIS EARLY ONSET OF
DISEASE.
SO SOME DISCUSSION THAT CAME UP
OF, YOU KNOW, DOES THE NOSE
REFLECT WHAT'S HAPPENING IN THE
DEEPER LUNG PARENCHYMA, CAN WE
ACTUALLY GO INTO THE BLOOD TO
PROFILE WHAT IS GOING ON AND SO
FORTH.
NEXT, WHAT KIND OF THERAPEUTIC
INTERVENTIONS ARE CURRENTLY
FUNCTIONING THROUGH TARGETING
PROGENITORS OR RESILIENCE.
WE'VE HEARD ABOUT SOME THINGS
THAT ARE REALLY ON THE CUTTING
EDGE, BUT JUST TAKING A STEP
BACK, ARE THERE THINGS THAT WE
REALLY ARE CURRENTLY DOING THAT
MAY ACTUALLY WORK ON THIS LEVEL
AND WE DON'T EVEN UNDERSTAND
JUST YET.
I THINK THAT RAVI'S INITIAL
POINT ON GENERATING AN ANALOGOUS
VIEW TO CARDIOVASCULAR HEALTH IS
SOMETHING THAT IS VERY IMPORTANT
TO DO.
TRYING TO DERIVE SOME OF THESE
EARLY BIOMARKERS THAT WE MAY BE
ABLE TO MEASURE THROUGH A SIMPLE
BLOOD DRAW OR PERHAPS AN EVEN
SIMPLER NASAL SWAB WOULD BE FON
TAS TICFANTASTIC IN TERMS OF TRG
THINGS REALLY ON A POPULATION
SCALE AND LASTLY I COME BACK TO
THE QUESTION OF -- WE'VE HAD A
LOT OF NIGHTS TALKS FROM BASIC
SCIENTISTS AND CLINICIANS BUT AT
THE END OF THE DAY THE
TRANSLATION INTO THE CLINIC
REALLY RELIES A LOT ON BIOTECH
AND INDUSTRY LAYER AND CLOSE
COLLABORATION BETWEEN ALL OF US
AND SO AGAIN JUST PROMPTING HERE
FOR ADDITIONAL DISCUSSION.
SO WITH THAT, I'LL PAUSE ON
THIS.
I KNOW I'VE ONLY HAD A BRIEF
CHANCE TO SUMMARIZE A LOT OF
BEAUTIFUL WORK THAT WAS
PRESENTED OVER THE PAST COUPLE
OF DAYS, BUT I HOPE THAT WE HAVE
SOME OPPORTUNITY TO DISCUSS SOME
FORWARD THINKING IDEAS HERE.
YOHANNES, PLEASE.
>> THANK YOU FOR PUTTING
TOGETHER THESE POINTS, JOSE.
THE ONE THING THAT I WOULD LIKE
TO MENTION IS OBVIOUSLY THERE
WERE A FEW DISCUSSIONS GOING ON
ABOUT POLYMORPHISMS OR
SUSCEPTIBILITY, AND I THINK THAT
IS, TO YOUR FIRST QUESTION, IS
RESILIENCE ABSENCE OF DISEASE OR
PRESENCE OF A SPECIAL STATE.
OR PROGENITOR CELLS, WHAT ARE
THEY DOING AND DO THEY HAVE
SUSCEPTIBILITY SOMEHOW WHEN THEY
ARE TRANSITIONING TO A STATE
THAT IS ACTUALLY REGENERATING
THE NORMAL LUNG.
I THINK THESE ISSUES ARE KIND OF
DIFFICULT FOR US TO CAPTURE AT
THIS POINT BECAUSE WE DON'T
UNDERSTAND THE BIOLOGY VERY
WELL, AND THERE IS A LOT OF WORK
BEING DONE THAT IS IMPRESSIVE.
AND IT'S GREAT, BUT I THINK
POLYMORPHISM IS ONE THAT SHOULD
BE CLEARLY PUT INTO THIS
EQUATION.
AND THE OTHER THING OBVIOUSLY IS
ALSO EXPOSURE, AND THAT'S
SOMETHING THAT IS VERY DIFFIC
DIFFICULT.
DATA ANALYSIS LIKE SAM SHOWED US
IS AN IMPORTANT ONE BUT DAVID
MENTIONED FOR EXAMPLE THAT
EXPOSURE NOT ONLY TO POLLUTION
PM2.5 WHICH IS BECOMING
EXTREMELY IMPORTANT NOW WITH
WILDFIRES RAGING ALL OVER THE
U.S. AND EVERYWHERE IN THE WORLD
REALLY, AND HOW TO CAPTURE THAT
NOT ONLY NOW FROM THE PM 2.5. OF
VIEW, BUT WE'VE SHOWN, FOR
EXAMPLE, PM2.5 IS NOT THE SAME
LIKE PM2.5 FROM WOOD SMOKE OR
FROM CIGARETTE SMOKE.
THEY CAN BE SYNERGISTIC, IN
FACT, IN CAUSING DISEASE.
SO THEY THERE ARE IN FACT A LOF
FACTORS THAT NEED TO BE PUT INTO
THE EQUATION TO START
UNDERSTANDING HOW TO GO FORWARD
AND DISENTANGLE, IF YOU WILL,
THE UNDERSTANDING OF THIS
RESILIENCE ISSUE.
BUT THE WORD ITSELF IS, I THINK,
APPROPRIATE AND CORRECT.
BUT THESE ARE JUST THE
COMPLEXITYS THAT JUST MAKE IT AT
THIS POINT REALLY DIFFICULT FOR
US TO CAPTURE EVERYTHING.
>> SO PERHAPS IF I CAN KIND OF
TIE WHAT YOUR COMMENT WAS AROUND
THE IDEA OF RESILIENCE, YOU'RE
PROPOSING, OF COURSE, THERE'S
DIFFICULTIES IN MEASURING WHAT
SOME OF THESE ENVIRONMENTAL
CHALLENGES ARE IN QUITE A
HOLISTIC WAY, BUT ALSO AT THE
SAME TIME, WE MAY WANT TO THINK
ABOUT DIFFERENCES IN ENVIRONMENT
THAT PEOPLE MAY NEED TO BE
RESILIENT FOR, AND IN ESSENCE,
THINKING ABOUT DISTINCT
ENVIRONMENTAL EXPOSURES, WE
THINK A LOT IN OUR LABORATORY
ABOUT VIRAL CHALLENGES, FOR
EXAMPLE, WE THINK LESS IS A
BLIND SPOT FOR US ABOUT
PM2.5 EXPOSURE.
SO I THINK THAT THINKING IF THIS
SAME MECHANISMS OR WHICH
MECHANISMS APLAY APPLY FOR WHIH
ENVIRONMENTAL CHALLENGES BROADLY
SPEAKING IS CERTAINLY SOMETHING
THAT WE NEED TO THINK A LOT
ABOUT GOING FORWARD.
>> YES, AND I MENTIONED PM2.5,
BUT CLEARLY ENVIRONMENT IS NOT
ONLY POLLUTION, RIGHT?
I MEAN, THERE IS FITNESS THAT
RAVI MENTIONED, FOR EXAMPLE, AND
SOCIAL STRESS.
I MEAN, THESE ARE ISSUES THAT WE
DON'T EVEN HAVE A FOOD HANDLE
ON, HOW TO QUANTIFY AND PUT INTO
THE EQUATION.
BUT ESSENTIALLY THIS
ENVIRONMENTAL ISSUE IS A VERY
DIFFICULT ISSUE AND SHOULD BE
CONSIDERED.
WE DIDN'T HEAR MUCH ABOUT
OXIDATIVE STRESS, WHICH IS ALSO
VERY DIFFICULT TO STUDY AND AS A
BASIC SCIENTIST WE KNOW STRESS
IS REALLY A COMPLICATED ISSUE AS
WELL.
SO THERE ARE THINGS LIKE THAT
THAT WE DIDN'T EVEN TOUCH UPON
IN THIS SESSION.
>> I THINK AS SOME OF THE
CAMERAS TURN ON HERE, I KNOW
THAT DOING THIS KIND OF VIRTUAL
ROUNDTABLE IS NOT AS NATURAL AS
DOING SO IN PERSON, BUT PLEASE,
I CERTAINLY DON'T NEED TO BE THE
NEXUS IN COMMUNICATION HERE.
SO SAM, IF YOU WANT TO CALL OUT
ON SOMEONE ELSE, OR DAVID I SEE
YOUR HANDS ARE UP, LET'S TRY TO
STIMULATE SOME INTERACTIONS HERE
JUST LIKE THE DIAGRAMS THAT YOU
WERE SHOWING, SAM, AS WE START
TO LAYER IN SOME ADDITIONAL
COMPLEXITY.
SAM, I THINK YOU WERE NEXT.
>> SURE.
AND DAVID, YOU CAN GO AFTER ME.
I THINK IT'S A GOOD FRAMING TOO.
I JUST WANT TO TALK ABOUT THIS,
I'VE BEEN THINKING ABOUT THIS, I
DIDN'T REALIZE HOW MUCH THERE
WAS GOING TO BE A THESIS ABOUT
RESILIENCE HERE, AND I THINK IT
IS VERY INTERESTING AND I THINK
THE WAY YOU'RE FRAMING IT
ACTUALLY WITH THESE TWO
DIFFERENT DEFINITIONS IS QUITE
APPROPRIATE.
IN MY HEAD WHEN I STARTED
HEARING IT, I WAS THINKING
SYSTEM STABILITY.
LIKE IN DYNAMICAL SYSTEMS THEORY
AND SYSTEMS BIOLOGY, PEOPLE TALK
A LOT ABOUT STABLE POINTS OF A
SYSTEM.
AND THEY'LL THINK ABOUT, IS
THERE A STABLE POINT IN A
CERTAIN CONFIGURATION OR ISN'T
THERE?
WHEN YOU BRING IN THIS CONCEPT
OF RESILIENCE, I THINK IT'S
QUITE MATHEMATICALLY
WELL-DEFINED BECAUSE YOU CAN SAY
SORT OF HOW DEEP IS THAT ENERGY
WELL, HOW STABLE IS IT, HOW
MUCH -- HOW HARD IS TO KICK IT
OUT OF HOMEOSTASIS OR TO MOVE
THAT SYSTEM.
YOU CAN ALSO TALK ABOUT THAT
ELASTICITY.
HOW FAR AWAY FROM A GIVEN STABLE
POINT, HOW MUCH DO YOU HAVE TO
PERTURB THE SYSTEM AWAY FROM
THAT PLACE BEFORE YOU GO OVER A
SADDLE AND YOU'RE INTO A
DIFFERENT ENERGY WELL WHICH WE
CAN THINK ABOUT TRANSITION TO
FIBROSIS.
HOMEOSTASIS INFLAMMATION OR ALL
THE WAY TO FIBROSIS.
SO I DO THINK THE WORD HAS THE
NECESSARY MEANINGS WITHIN IT AND
I THINK IT IS QUITE APPROPRIATE.
DAVID.
>> I HAVE A COUPLE OF COMMENTS,
BUT I'LL RESERVE MAY COMMENTS
ABOUT THE ENVIRONMENT FOR LATER
BECAUSE I THINK THAT THEY MAY
LEAD INTO SORT OF FUTURE
OPPORTUNITIES.
IN TERMS OF THE CONCEPT OF
RESILIENCE, I'VE BEEN THINKING
FOR THE PAST TWO DAYS ABOUT IT
BEING SO CONTEXT-DEPENDENT.
CONTEXT DEPENDENT IN TERMS OF
AGE, STAGE OF DISEASE, A VARIETY
OF DIFFERENT FACTORS THAT ARE
GOING TO AFFECT THE INHERENT
RESILIENCE AND THE INHERENT
RESILIENT STATES WHEN CHALLENGED
AND STRESSED IN CERTAIN WAYS.
SO I THINK I AGREE WITH WHAT SAM
IS STAYING, IT'S COMPLEX AND THE
TRANSITION STATES ARE REALLY
SOMEWHAT DEPENDENT ON THE POINT
IN TIME THAT THEY'RE BEING
ASSESSED, AND IN THAT CONTEXT, I
THINK IT'S VERY DANGEROUS TO
GENERALIZE BEYOND THAT POINT AND
CONTEXT.
SO I THINK THAT WE NEED TO SORT
OF BEGIN TO THINK ABOUT HOW TO
INTEGRATE THESE FINDINGS ACROSS
THESE DIFFERENT CONTEXTUAL
DISEASE AND DEMOGRAPHIC AND
EXPOSURE STATES AS A WAY OF
UNDERSTANDING THE PLASTICITY OF
RESILIENCE.
>> CAN I ADD ONE SENTENCE TO ADD
TO WHAT YOHANNES MENTIONED
EARLIER.
THE GENE ENVIRONMENT, THE GWAS
FIND DIFFERENT VARIANTS OF --
IPF BUT THOSE VARIANTS THEY HAVE
TO KIND OF DETERMINE DIFFERENT
VARIANCES -- WHEN THEY SEE THE
SMOKER THAT DEVELOPS COPD.
SO WHEN WE TALK ABOUT RESILIENCE
OF CELL TYPE, I THINK THESE
GENETIC COMPONENTS HAVE TO BE
CONSIDERED CAREFULLY, ESPECIALLY
IN RESPONSE TO THE ENVIRONMENT
OF CHALLENGE.
THAT'S WHERE THE GENETIC
DETERMINANTS ACTUALLY PLAY A
ROLE, HOW TO STRESS THE -- WHEN
YOU SEE BIG DIFFERENCE BETWEEN
THESE TWO GROUPS OF ALLELE
CARRIER.
ABOUT THIS GENETIC -- SO THE
GENE PROBABLY PLAYS A ROLE IN
DIFFERENT CELL TYPE AS WE HAVE
HEARD IN THE DIFFERENT -- LIKE
FIBROBLASTS AND IMMUNE CELL
TYPE.
>> RAVI, I KNOW PERHAPS YOU HAD
SOME THOUGHT, I THINK YOU MIGHT
HAVE USED EVEN THE WORD TIPPING
POINT INSTEAD OF SADDLE OR
SOMETHING LIKE THIS IN TERMS
OF -- BUT PERHAPS IF WE CAN KIND
OF SCALE BACK OUT TO THE PATIENT
OR EPI LEVEL RATHER THAN THE
KIND OF PURE SYSTEMS DATA LEVEL,
AS WHAT SAM WAS MENTIONING, HOW
ARE YOU KIND OF VIEWING THIS OR
INTERPRETING THIS PROBLEM?
>> YEAH, I MEAN, I THINK THIS IS
KIND OF THE CHALLENGE THAT WE'RE
FACED WITH, IS THAT WE ARE
PRECONDITIONED BY OUR TRAINING,
FUNDING MECHANISMS AND
MOTIVATIONS TO STUDY DISEASE.
THAT'S OUR BUSINESS MODEL.
AND YET, IF WE WANT TO CONFRONT
PUBLIC HEALTH PROBLEMS, WE HAVE
TO STUDY HEALTH.
NOT JUST DISEASE.
THE MINORITY OF PEOPLE DEVELOP
DISEASE BUT ALSO THE MINORITY OF
PEOPLE REMAIN HEALTHY THROUGHOUT
THEIR LIFESPAN.
THAT'S SORT OF ALL THESE IN
BETWEEN POINTS.
IN THE RESPIRATORY SYSTEM, THERE
ARE AN ENDLESS POINT OF INJURY,
LIKE YOHANNES WAS SAYING.
SOME PEOPLE HAVE AIR POLLUTION,
SOME PEOPLE SMOKE, SOME PEOPLE
LIVE IN MOLDY HOMES, SOME PEOPLE
ARE SENSITIVE TO ALLERGENS, SOME
PEOPLE GET VIRAL INFECTION OVER
AND OVER AGAIN AND RECOVER, SOME
PEOPLE GET VIRAL INFECTIONS THAT
SEEM TO HAVE UNRELENTING DECLINE
AFTERWARDS.
AND THAT HETEROGENEITY OF WHO
CONFRONTS VARIOUS TIPPING POINTS
ADVAIR JUST TIMES, SORT OF WHAT
DAVID WAS JUST SAYING, AND TIP
TOWARDS AN UNRELENTING DECLINE,
VERSUS HAVE AN IMPAIRMENT FOR A
SHORT PERIOD OF TIME, THEN ARE
ABLE TO HEAL, RECOVER, VERSUS
THOSE WHO SEEM UNAFFECTED BY ALL
OF THESE THINGS NO MATTER WHAT
THEY DO THROUGHOUT THEIR LIFE,
ARE REALLY IMPORTANT QUESTIONS
TO UNDERSTAND HEALTH.
SO I UNDERSTAND THE VEDZ WHO
HAVE IDEAL RESPIRATORY HEALTH
AND WHY THEY DO OR DON'T RESPOND
TO INJURY.
UNDERSTANDING THOSE WHO DEVIATE
FROM HEALTH BUT ARE ABLE TO GET
BACK TO HEALTH IS REALLY
IMPORTANT, AND I THINK WE'VE
HEARD A LOT OF INFORMATION ABOUT
PEOPLE WHO DESEND INTO SORT OF
DISEASE AND VARIOUS MECHANISMS
AROUND IT, BUT I THINK I WOULD
ENCOURAGE US TO SORT OF
CHALLENGE THE THINKING A LITTLE
BIT AROUND HOW TO STUDY AND
PROMOTE HEALTH VERSUS SORT OF
THIS UNRELENTING DECLINE TOWARDS
DISEASE.
NOW OF COURSE I DON'T KNOW HOW
TO DO ANY OF THAT, BUT THAT'S
SORT OF, I THINK, THE CHALLENGE
WE'RE FACED WITH.
WELLINGTON, PLEASE, YOUR
INSIGHTS ARE ALWAYS GOOD SO GO
AHEAD.
>> THANK YOU.
TALKING ABOUT RESILIENCE, THERE
IS ONE FACTOR THAT WE PROBABLY
VHAVE TALKED ABOUT A LITTLE BIT,
BUT I'D LIKE TO EMPHASIZE THAT
IT IS THE FACTOR AGE, AGING,
BECAUSE RESILIENCE HAS A LOT TO
DO WITH AGE AND ONE OF THE
THINGS THAT ARE ACTUALLY EVEN
MORE COMPLICATED IS TO DEFINE
WHAT IT IS ENVIRONMENT VERSUS
WHAT IT IS AGING, AND I THINK IT
IS PERHAPS WORTH AT LEAST
GETTING A LITTLE BIT INTO THIS,
ALTHOUGH THIS IS A VERY, VERY
COMPLEX SUBJECT.
>> YEAH, I CAN'T AGREE WITH YOU
MORE, WELLINGTON.
I THINK AGE IS A HUGE ISSUE, AND
THE DYNAMICS THAT OCCUR IN TERMS
OF AGING, IN TERMS OF RESILIENCE
WILL TEACH US SO MUCH ABOUT
RESILIENCE.
>> ANDREW AND SUSAN ARE HAVING A
CONVERSATION IN THE CHAT WITHOUT
US, BUT I WOULD ENCOURAGE TO YOU
SHARE, ANDREW OR SUSAN.
>> SUSAN, DO YOU WANT TO GO
FIRST?
SORRY, GO AHEAD.
>> SORRY.
JUST TRYING TO WEIGH IN A LITTLE
BIT.
THERE ARE SOME GOOD
EPIDEMIOLOGIC STUDIES OUT THERE
ABOUT NOT NECESSARILY IN THE
LUNG FIELD ABOUT HE EXTREME
PHENOTYPES, AND I JUST WONDER IF
WE NEED TO FOSTER WAYS OF DOING
THOSE KINDS OF STUDIES, WHETHER
IT'S ON A GWAS SCALE OR WHAT, TO
BE ABLE TO IDENTIFY THOSE
INDIVIDUALS WHO ARE 40 PACK A
YEAR SMOKERS BUT STILL HAVE
REALLY GOOD LUNG FUNCTION.
WHAT ARE THOSE EXTREME
PHENOTYPES THAT REALLY MAKE YOU
THINK ABOUT THIS INDIVIDUAL HAS
SOME SORT OF GENETIC BACKGROUND
THAT PROMOTES RESILIENCE.
>> I CAN JUST ADD ON TO THAT
MAYBE.
I TOTALLY AGREE WITH THAT
COMMENT, AND IT MAKES ME THINK
ABOUT LONGITUDAL COHORT IN WHICH
THE PARTICIPANTS HAVE BEEN
CAREFULLY PHENOTYPES AND WE KNOW
THEIR GENETIC BACKGROUND AS WELL
AS WHAT EXPOSURES THEY HAD.
AN EXAMPLE I HAD IN MIND
ALTHOUGH THERE ARE CERTAINLY
OTHERS IS THE FRAMINGHAM HEART
STUDY WHERE INDIVIDUALS HAVE
BEEN CHARACTERIZED GENETICALLY,
POLYGENIC RISK SCORES TO THINK
ABOUT THEIR RISK FOR VARIOUS
DISEASES AND THEN SPECIFICALLY
IN THE CASE OF FRAMINGHAM, WE
COLLECTED REPROGRAMMABLE BLOOD
CELLS ON THE ENTIRE EXISTING
COHORT SO WE HAVE THE PRECURSOR
CELLS TO MAKE THE IPSC CELLS IF
WE WANT TO ON OVER 6500
FRAMINGHAM PARTICIPANTS AND YOU
CAN ALSO IMAGINE COLLECTING
OTHER TISSUES FROM THOSE
PARTICIPANTS, AND I THINK YOU
COULD SIMILARLY APLAY THAT
PARADIGM TO THE LUNG HEALTH
STUDY, FOR INSTANCE, THAT'S
CURRENTLY ENROLLING OR TO OTHER
LONGITUDAL COHORT THAT WOULD
ALLOW YOU TO IDENTIFY THESE
EXTREME -- LIKE SUSAN MENTIONS.
>> THIS IDEA, ANDREW, OBVIOUSLY
WE WERE EMAILING ABOUT THIS
EARLIER THIS WEEK SO IT'S
TIMELY, THIS IDEA OF TAKING
ADVANTAGE OF THE
POPULATION-BASED COHORTS, THE
EPI STUDIES LIKE FRAMINGHAM OR
CARDIA, NOT JUST BECAUSE I WORK
WITH IT BUT BECAUSE IT'S
ACTUALLY A GOOD EXAMPLE, BUT
WHERE WE COULD CREATE IPSC
LIBRARIES FROM INDIVIDUAL
PEOPLE, RIGHT, WHO WE KNOW THEIR
LIFECOURSE ALREADY.
IT DOES AFFORD A HUGE
OPPORTUNITY, I THINK, TO
UNDERSTAND THESE MECHANISMS AND
LINK THEM BACK TO SORT OF WHAT
WE ALREADY KNOW ABOUT THE
LIFECOURSE OF THESE INDIVIDUALS
AND SORT OF THE AIRWAY CELLS AND
THE ALVEOLAR CELLS AND SORT OF
ALL THESE THINGS, IT SEEMS LIKE
A VERY, VERY COMPELLING
OPPORTUNITY TO ME.
>> JUST TO ELABORATE A TINY BIT
FURTHER, YOU CAN FIND
INDIVIDUALS WITHIN FRAMINGHAM,
FOR EXAMPLE, WHO HAVE 30 PACK A
YEAR HISTORY OF SMOKING AND HAVE
NORMAL PFTs AND DON'T HAVE
SIGNIFICANT EMPHYSEMA ON A CT
SCAN SO I WOULD HYPOTHESIZE
THOSE WOULD BE THE GENETICALLY
RESILIENT INDIVIDUALS WHO CAN
UNDERGO THESE ENORMOUS EXPOSURES
AND NOT DEVELOP DISEASE.
>> AND IMPORTANTLY IN FRAMINGHAM
AND CARDIA, IT'S NOT JUST
DEVELOPING EMPHYSEMA.
WITH CT TECHNOLOGY WE CAN
MEASURE EVEN INTERMEDIATE --
WHAT LOOKS LIKE INJURED LUNG AND
FIVE YEARS LATER, IT'S PRISTINE
AGAIN.
I THINK THESE ARE REALLY
IMPORTANT THINGS TO CONSIDER.
>> I WOULD LIKE TO ACTUALLY -- I
FIND THIS VERY, VERY, VERY
IMPORTANT, RAVI, WHAT YOU HAVE
ACTUALLY MENTIONED, AS A WAY TO
APPROACH THIS QUESTION, WHICH IS
VERY DIFFICULT AS I HAVE
MENTIONED.
NOW I WOULD LIKE TO INTRODUCE
ANOTHER QUESTION THATS IS R. IT-
WHEN YOU TALK ABOUT RESILL
YERNTION THIS IS A LUNG
WORKSHOP, BUT IS RESILIENCE THE
SAME IN ALL ORGANS OF AN
INDIVIDUAL?
SO HERE WE CAN ACTUALLY HAVE
SOMETHING THAT IT IS -- WOW,
THIS PERSON LIVED FOR 103 YEARS
OLD OR 90-SOMETHING, IS THIS A
CONJUNCTION OF THINGS, IN FACT,
THAT WHAT WE HAVE IS A
RESILIENCE OF EACH INDIVIDUAL
ORGAN ALL TOGETHER OR IS THERE
SOMETHING THAT BIASED US
TOWARDS -- AGAIN, PERHAPS THE
IPSC AS AN APPROACH TO GO OVER
THIS IS AN INTERESTING ONE,
BECAUSE IT WOULD DEPEND ON YOU
OF COURSE DERIVE ALL OF THESE
DIFFERENT -- FROM HIM OR HER.
>> I THINK THAT'S A VERY
INTERESTING POINT THAT YOU'RE
RAISING.
THE OTHER THING THAT COMES TO
MIND IS OF COURSE PRIMARY
CELL-BASED MODELING RELATIVE TO
IPS, I MEAN, VERY DIFFERENT
CHALLENGE BUT ONE COULD ALSO
IMAGINE THAT THE LIFE HISTORY OF
ENVIRONMENTAL EXPOSURES, NOW WE
HAVE OVER THE PAST FOUR OR FIVE
YEARS INDIVIDUALS WILL BE
RECOVERING FROM VERY SEVERE
COVID INFECTIONS IN A WAY THAT
PERHAPS OTHER RESPIRATORY
VIRAL -- IS THIS GENETICS, YOU
KNOW, OR IS THIS IS NOT?
WE UNDERSTAND NOW, FOR EXAMPLE,
IT'S SUCH A HARD EXAMPLE TO WRAP
MY MIND AROUND BUT YOU WOULD
THINK HIGH LEVELS OF ACE 2 IS A
GOOD THING BECAUSE IT'S
TISSUE-PROTECTIVE IN STERILE
LUNG INJURY.
IT'S TISSUE-PROTECTIVE IN FLU
INFECTION.
HOWEVER, THIS MAY NOT BE THE
BEST THING IN TERMS OF
SARS-COV-2 AND COVID-19, SO I
THINK THAT, DAVID, YOU WERE THE
ONE WHO I THINK MENTIONED
SOMETHING ABOUT CONTEXT
DEPENDENCY OF RESILIENCY AND THE
THINK CHERRY PICKING ONE GENE
AND ONE TISSUE, HERE'S ONE
EXAMPLE WHERE IT COULD GO
MULTIPLE WAYS.
BUT THE THINK THE IDEA OF THE
I.IPSC KIND OF BIOBANK APPROACHS
IS QUITE INTERESTING TO GET AT
THE HEART OF SOME OF THESE
GENETICALLY GROUNDED IDEAS, BUT
BRINGING IN THE ENVIRONMENT IS
ALWAYS THE TENSION THERE.
>> JUST TO BUILD ON WHAT YOU
SAID, I THINK WE KNOW QUITE A
BIT ABOUT THE GENETICS OF
DISEASE.
WE DON'T KNOW NEARLY ENOUGH
ABOUT THE GENETICS OF
ENVIRONMENTAL SUSCEPTIBILITY.
AND WHEN YOU THINK ABOUT HOW
THAT RELATES TO RESILIENCE, IT'S
ABSOLUTELY CRITICAL.
AND SO I THINK THAT'S AN AREA
THAT REALLY DESERVES A LOT MORE
EMPHASIS OVER THE NEXT SEVERAL
YEARS TO FIGURE OUT WHAT MAKES
THOSE INDIVIDUALS RESILIENT TO
ENVIRONMENTAL STRESS, OR THE
FLIP SIDE IS WHY ARE SOME PEOPLE
SO SUSCEPTIBLE TO ENVIRONMENTAL
STRESS.
SEPARATE FROM DISEASE.
SEPARATE FROM DEVELOPMENTAL
DISEASE.
>> AND THIS IS ACTUALLY -- I
WANT TO CHIME IN, IS THAT MAYBE
NEURON PLAY A ROLE IN THIS
ADAPTATION COULD SOME WAY IN THE
FUTURE WOULD DEVELOP TOOLS TO
JUST STIMULATE THE CERTAIN TYPE
OF NEURONS IN VAGUS NERVE TO
TREAT, FOR EXAMPLE, ASTHMAS OR
EVEN PROMOTE -- BECAUSE IN THE
VEGAS NERVE THERE'S SO MANY
NEURONS BUT IF WE COMBINE WITH
SINGLE CELL -- IDENTIFY
INDIVIDUAL NEURONS AND THEN
INTERFERE CERTAIN TYPES OF
NEURONS FOR TREATMENT AND ALSO
FOR ADAPTATIONS, AND I THINK
THAT WOULD IMPROVE RESILIENCE
FOR HUMANS AS WELL IN THE
FUTURE.
>> I JUST WANT TO TAKE A MINUTE
TO CHIME IN ABOUT THE VALUE OF
USING THE FULL EXTENT OF THE
LIFECOURSE FOR THESE SORTS OF
STUDIES, BECAUSE WE'RE FACED IN
THE NEONATAL INTENSIVE CARE UNIT
WITH A DELLS OF PAT OODLES OF PT
UNDERGO THE SAME ENVIRONMENTAL
EXPOSURES AND YET HAVE VERY
DIFFERENT OUTCOMES THAT ALSO
IMPAIRED THEIR ABILITY TO
ACHIEVE MAXIMUM LUNG GROWTH
POTENTIAL.
AND I THINK DOING SOME OF THESE
STUDIES IN THOSE POPULATIONS IS
CRITICAL.
>> PICKING OUT THE POPULATIONS
WHERE YOU HAVE THOSE EXTREME
EXPOSURES IS A GREAT WAY OF
DOING THE WORK.
>> I THINK ONE CHALLENGE THAT
CAME TO MIND IS THIS QUESTION OF
HOW REGIONALIZED SOME OF THESE
RESPONSES ARE.
WE SAW SOME VERY BEAUTIFUL
IMAGES EITHER OF JUST ONE
EPITHELIAL LAYER AND ALVEOLUS,
CLUSTER OF DIFFERENT ALVEOLI,
THE WHOLE LUNG, AND THERE'S
REGIONALITY IN ALL OF THE
DIFFERENT RESPONSES.
THERE'S EVEN SOME AREAS OF THE
LUNG AND OTHER AREAS THAT LOOK
TOTALLY FINE OR TWO DISEASES IN
THE SAME LUNG IN SOME CASES, SO
I'M WONDERING, GIVEN THAT'S
CHALLENGES, HOW DO WE THINK
ABOUT REGIONALITY IN TERMS OF
KIND OF NEXT GENERATION
THERAPEUTICS.
>> ARE YOU TALKING ABOUT THE
STEM CELLS IN DIFFERENT REGIONS?
>> STEM CELLS IN DIFFERENT
REGIONS OR JUST EVEN WITHOUT
NECESSARILY NEEDING TO INVOKE
THE STEM CELLS, I THINK WE SEE
IN MANY OF THE DISEASES THAT
THERE'S CERTAIN AREAS OF THE
LUNG THAT WILL BE QUITE HIGHLY
AFFECTED AND OTHER ONES WHICH
SEEM TO BE COMPLETELY NORMAL IN
TERMS OF ARCHITECTURE.
AND SO OBVIOUSLY THIS POSES A
CERTAIN CHALLENGE IN A WAY, YOU
KNOW, IT'S NOT LIKE THE WHOLE
TISSUE IS DISEASED, SOME OF IT
IS OKAY, SOME OF IT IS NOT, SO
THIS IS A CHALLENGE IN BOTH
UNDERSTANDING THE SYSTEM BUT
ALSO WHEN IT COMES TO THINKING
ABOUT THE THERAPEUTIC OPPORTUNI.
CARLA, DID YOU WANT TO MENTION
SOMETHING?
>> YES, THANK YOU.
THIS IS CARLA KIM.
FIRST THANK YOU ALL FOR
ORGANIZING THIS AND IT'S A
PLEASURE TO BE INCLUDED.
I WAS WANTING TO POTENTIALLY
HAVE US THINKING ABOUT SYSTEMIC
INFLUENCES AND MAYBE A LITTLE AT
WHAT YOU'RE TOUCHING ON OF HOW
MEASURING CHANGES IN ONE PART OF
THE PULMONARY SYSTEM COULD BE
REFLECTIVE OF OTHER CHANGES.
WE'VE BEEN STUDYING HOW AIRWAY
INJURY IMPACTS THE ALVEOLAR
CELLS AND HOW THAT COULD BE
MEDIATED THROUGH IMMUNE CELL
SIGNALING.
SO I WAS THINKING ABOUT THIS
YESTERDAY, MAYBE IT WAS EARLIER
TODAY WHEN DR. GOCHE WAS TALKING
ABOUT CAN THE NASAL CELLS TELL
US WHAT'S GOING ON DEEPER IN THE
LUNG.
WE DON'T KNOW MUCH ABOUT THAT.
SO THE KIND OF STUDIES WHERE WE
CAN PURPOSELY COMPARE CELLS FROM
DIFFERENT REGIONS, FROM THE SAME
PERSON OR SYSTEM, AND GET A
HANDLE ON A MUCH BROADER SCALE
OF WHAT ARE THE CHANGES GOING
ON.
THAT COULD REVEAL OPPORTUNITIES
WHERE -- FOR DETECTION AND ALSO
FOR TREATMENT.
I'M ALSO SITTING HERE AT AN
EARLY STAGE LUNG CANCER
DETECTION CONFERENCE, AND I
THINK THERE'S A LOT OF ROOM
WHERE OUR FIELD, YOU KNOW, OFTEN
THROUGH FUNDING MECHANISMS WE
DON'T HAVE A PROJECT THAT
INVOLVES CANCER WHEN IT'S NHLBI,
BUT I THINK THERE'S A LOT OF
PRECANCEROUS SITUATIONS THAT
WE'RE ALL VERY CURIOUS OF HOW
MUTATION OR EXPOSURE MIGHT ALTER
SUSCEPTIBILITY FOR A LUNG
DISEASE OR LUNG HEALTH AGING
THAT PERHAPS IS POTENTIALLY
GOING ALONG WITH PRECANCEROUS
CONDITIONS.
SO MAYBE WAYS OF CONNECTING
ACROSS AGENCIES ALSO, SUCH AS A
JOINT EFFORT BETWEEN NIA, NCI
AND NHLBI SO WE CAN ASK SOME OF
THESE TYPES OF QUESTIONS.
SO THAT WAS A LOT, BUT JUST SOME
THOUGHTS THERE.
>> THIS IS MOUMITA.
I THINK CARLA WAS JUST TALKING
ABOUT THE PRESENTATION THAT I
DID YESTERDAY, AND WE DO SATISFY
UPPER AIRWAY PROGENITORS
ACTUALLY CORRELATED WITH BOTH
LUNG FUNCTION DECLINE AND
DIFFUSION ISSUE.
WE DO SEE THAT SAME WITH NASAL
BRUSHINGS, SO ARE THESE FOR
IMMUNE CELLS, FOR SOME KIND OF
NEURONAL CONNECTION WE HEARD
TODAY, HOW ARE CELLS
COMMUNICATING.
WE KNOW THAT THERE ARE REGIONAL
PROGENITORS BUT CLEARLY THERE IS
SOME KIND OF A UNIFIED THEME.
COULD THAT BE THAT THIS
COMMUNICATION IS VIA IMMUNE
SYSTEM, COULD THAT BE VIA SOME
KIND OF COMMUNICATION WITH
NEURON SYSTEM?
AND I CANNOT, LIKE, YOU KNOW, --
PREMALIGNANCY, PRECANCER
STANDPOINT BECAUSE WHEN WE FIRST
STARTED LOOKING AT IT, THAT
PRETTY MUCH THE PREMALIGNANCY
POPULATION IS ABOUT 30 TO 40% OF
PEOPLE ALREADY HAVE COPD.
THEY'RE HEAVY SMOKERS.
ALSO WE FOUND PEOPLE, THEY ARE
LIKE IN THEIR 70s, WITH
HUNDRED PACK YEARS SMOKING
HISTORY, BUT THEY DON'T HAVE
COPD.
THEY MAY HAVE THAT PRE-COPD
PHENOTYPE WHICH HAS -- WHICH
DOESN'T EXIST ANYMORE SO YES,
THEY COULD BE AT RISK, BUT AT
THIS POINT THEY'RE REALLY A
RESILIENT POPULATION RATHER THAN
A SUSCEPTIBLE POPULATION.
SO HOW ARE THEY DIFFERENT WITH A
45-YEAR-OLD HAVING MUCH LESS
SMOKING HISTORY WITH LOW
NORMAL -- 76 BUT 100 PACK YEARS,
SO THE HETEROGENEITY BETWEEN
PATIENTS IS JUST SO DIVERSE,
THAT IT'S HARD TO MODULATE.
LIKE ANY MODEL SYSTEM, UNLESS WE
GO AND TRY TO MAKE IPC THAT WE
WERE DISCUSSING BEFORE.
>> DANIEL, WOULD YOU LIKE TO
ADD?
>> SO THIS IS JUST A FASCINATING
DISCUSSION.
BUT I WANTED TO GO BACK AND HAY
LIGHT SOMETHING ABOUT THE
ENVIRONMENT.
BECAUSE IF WE'RE TRYING TO STUDY
THE BEHAVIOR OF ANY GIVEN
PROGENITOR CELL, ITS HOME IS
CRITICALLY IMPORTANT.
SO THIS BECOMES A QUESTION OF
TOOLS.
AND SO IF YOU'RE TRYING TO STUDY
SOMETHING ON ALI OR ROUTINE
TISSUE CULTURE OR IN A GEL, A
GOOD STRATEGY IS TO TRY AND
MIMIC AS BEST POSSIBLE WHAT THIS
CELL IS GOING TO SEE IN REAL
LIFE.
AND SO YOU CAN TAKE THAT AND
SAY, WELL, IF I HAVE BASAL
EPITHELIAL CELLS OR WHATEVER
TYPE FROM A COPD PATIENT AND TO
HIGHLIGHT WHAT WAS SAID FOR
MULTIPLE COPD PATIENTS BECAUSE
YOU NEED THAT INDIVIDUAL
HETEROGENEITY AND VARIABILITY,
WHO ARE YOU GOING TO CULTURE
THEM ON?
MATRI GEL OR EXTRACELLULAR
MATRIX DERIVED FROM A NORMAL
LUNG OR A LUNG WITH EMPHYSEMA?
WHAT PART OF THAT LUNG ARE YOU
GOING TO CULTURE IT ON?
BECAUSE THE AIRWAYS ARE
DIFFERENT FROM THE ALVEOLI,
DIFFERENT FROM THE VASCULATURE.
THEY HAVE DIFFERENT MATRIX
PROTEIN COMPOSITION, DIFFERENT
GLYCO -- MATRIX ASSOCIATED
GROWTH FACTOR AND SO ON.
THIS ISN'T REALLY THAT DIFFICULT
A CONCEPT, BUT IF WE CAN
INCORPORATE THAT TYPE OF
THINKING INTO THE TOOLS THAT WE
UTILIZE, AND THEN YOU CAN GET
MORE USEFUL INFORMATION, THROW
IN OTHER FACTORS LIKE SICKLY CYC
MECHANICAL STRAIGHT, SO THROWING
STRETCH, HAVE YOUR CALL FOR
SYSTEM, STRETCH SOMEHOW.
AND YOU CAN HAVE VERY FREQUENCY
AND AMPLITUDE AND MIMIC NORMAL
VENTILATION THAT 8 TO 12, 12 TO
16 BREATHS A MINUTE VERSUS A BAD
EMPHYSEMA, 4 TO 6 BREATHS A
MINUTE OR WHATEVER.
SO AGAIN, THINKING MORE IN A
GRANULAR WAY IS GOING TO BUY US
A LOT MORE INFORMATION AND IT
WILL HELP DEVICE THEN I THINK
MORE TARGETED STRATEGIES.
IF YOU HAVE A PROGENITOR CELL,
TAKE FRANK AND SIMILAR WORK AND
YOU WANT TO PUT A PROGENITOR
CELL INTO A LUNG WITH EMPHYSEMA
FOR EXAMPLE WITH THE GOAL OF
STRUCTURALLY REGENERATING DISTAL
ARRAYS AND ALVEOLI, THAT CELL
NEEDS TO RESPOND TO WHAT IT'S
GOING TO SEE IN THAT DISEASED
LUNG WITH EMPHYSEMA AND
HOPEFULLY UTILIZE THOSE CUES TO
DO WHAT YOU HOPE IT GOING TO DO.
SO WHEN YOU LOOK IN FORWARD
LOOKING DIRECTIONS, THIS IS, I
THINK, AN IMPORTANT PLATFORM.
>> YOU THINK THAT'S A GREAT
POINT.
JUST FOLLOWING UP THE POINT THAT
WAS BROUGHT UP BY MULTIPLE
PEOPLE ABOUT THE GENE
ENVIRONMENTAL INTERACTION, THE
WAY I WANT TO -- ONE THING I
WANT TO ADD ON THAT POINT IS
THAT AS YOU SAID, MULTIPLE --
IT'S ALL ABOUT THE KIND OF THE
CONTEST DEPENDING ON THE STATE
OF YOUR LIFE AND THEN AGE AND
SOME CONDITIONS AND HOW YOU
REACT TO THE ENVIRONMENT
CONDITION AND THEN HOW YOU REACT
COULD BE ALREADY WIRED BY
GENETIC CONDITION THAT YOU
ALREADY HAVE BUT AT THE SAME
TIME, IT COULD BE THROUGH THE
METABOLISM, BECAUSE METABOLISM
COULD BE SOMETHING THAT YOU --
LIKE REACT TO THE EXTERNAL OR
INTERNAL STIMULI AS SHAWN DAVIS
SHOWED IN HIS FIRST TALK.
SO LIKE PREVIOUSLY PEOPLE
THOUGHT THAT LIKE A
TRANSCRIPTION FACTOR IE GENE
CONTROLLED THE M METABOLISM AND
DICTATE WHAT CELLS TO DO, BUT
THE WAY THE RECENT THEME KIND OF
THINKING IS THAT THE METABOLISM
ACTUALLY DICTATES THE
TRANSCRIPTION FACTORS, AND THEN
THE CONTROL, THE CELL BEHAVIOR.
SO IT KIND OF MAKES SENSE
BECAUSE LIKE THE METABOLISM CAN
BE INFLUENCED BY A LOT OF
EXTERNAL STIMULI, IE ENVIRONMENT
LIKE NUTRIENT AND STRESS AND ET
CETERA, AND JUST TELL THE CELL
THIS IS NOT THE TIME THAT YOU
REACT IN A CERTAIN WAY OR NOT.
AND THEN BASED ON THAT CELL
BEHAVES AND THEN UNTIL THOSE
STRESS OR EXTERNAL STIMULI KIND
OF SETTLE DOWN, THAT'S SORT OF
LIKE A CHECK-IN POINT, AND THE
SAME WAY, THAT'S HOW I ALSO
THINK ABOUT THE STRESS RESPONSE.
LIKE THE CHANGES.
THE CELLS HAVE TO HAVE A
CHECKPOINT BEFORE YOU COMMIT IT
INTO CERTAIN CELL BEHAVIOR SUCH
AS PROLIFERATION, WHATEVER, AND
THEN YOU JUST WAIT AND MAKE SURE
THAT YOU HAVE ALL THE CONDITION
APPROPRIATELY OR AT LEAST IN
CERTAIN CONTEXTS, THAT SHOULD --
WHATEVER THE APPROPRIATE
RESPONSE THAT THE CELL IS
COMMITTED.
AND UNTIL THOSE SETTLE DOWN,
THEN YOU JUST COMMITTED IN THE
RIGHT PATHWAY.
SO FROM THAT ASPECT, I THINK THE
ONE THING THAT WE NEED TO THINK
ABOUT IS THAT WE HAVE A VERY
WONDERFUL, VERY -- 3D CULTURE
SYSTEM AND A LOT OF -- VERY
VALUABLE IN VITRO SYSTEM, BUT AS
WE LEARN FROM THE CANCER
METABOLISM FIELD, THE INDIVIDUAL
METABOLISM IS VERY DIFFERENT
FROM IN VIVO METABOLISM, AND
SOME OF THE DISCREPANCY BETWEEN
THE ANIMAL STUDIES VERSUS THE
HUMAN CLINICAL TRIAL, I'M NOT
SAYING THIS IS ALL BUT PARTLY
COMING FROM THE STUDY FINDING,
IN VITRO STUDY FINDING AND W WE
KNOW IT'S VERY DIFFERENT, SO
THIS IS SOMETHING THAT WE NEED
TO JUST KEEP IN MIND, THAT THE
IN VITRO METABOLISM ITSELF IS
VERY DIFFERENT FROM IN VIVO, AND
IT COULD MAKE THE CELLS BEHAVE
DIFFERENTLY FROM WHAT ACTUALLY
CELLS BEHAVE INA VERY VO, WHICH
WE ACTUALLY CARE ABOUT.
SO THAT'S JUST THE ONE THING
THAT I'D WANT PEOPLE TO THINK
ABOUT.
>> I THINK IT TOOK A WHILE TO
LEARN THAT IN CANCER METABOLISM.
IT STARTED OFF AS ONE ONCOGENE,
ONE METABOLIC PHENOTYPE.
A LOT OF NICE WORK DONE EARLY IN
2010, MARIA BEING ONE OF THEM,
SHOWING YOU HAVE THE SAME
MUTATION IN TWO DIFFERENT PLACES
AND YOU HAVE COMPLETELY
DIFFERENT METABOLIC PHENOTYPES,
METABOLIC DEPENDENCIES, SO I
COMPLETELY AGREE, I THINK THAT
IS A -- METABOLISM ISN'T A
TWO-WAY STREET, IT'S NOT A
RESULT OF AN UPSTREAM SIGNALING
PATHWAY.
IT'S A RECIPROCAL PROCESS.
I HOPE THAT OUR TECHNOLOGY GETS
TO ANSWER SOME OF THOSE
QUESTIONS IN THE RIGHT MODELS.
>> I THINK WE CAN'T HEAR YOU AT
THE MOMENT.
>> I WANTED TO BRING UP A LITTLE
DIFFERENT TAKE ON RESILIENCE,
NOT A POPULATION LEVEL OR PERSON
LEVEL BUT AT A MOLECULAR LEVEL
AS TO WHERE I THINK YOU COULD
ARGUE FOR SOME MAYBE
ASPIRATIONALLY FOR HOW WE COULD
IMPROVE RESILIENCE.
ONE WOULD BE ABLE TO REGULATE
AND STABILIZE CONTROL AND TO
SOME DEGREE EP GENETIC STATES.
I MEAN, THE WHOLE FIELD OF
IPSLs IS HINGED ON THE GETTING
THE CELL RIGHT, BUT WE DON'T
REALLY CONTROL THE EPIGENETICS
AND IT'S SO EASY TO GO WRONG
THAT I THINK IN NATURAL --
NORMAL CELLS AND AN ORGANISM BUT
ALSO IN A CULTURE SYSTEM, I
THINK THERE SHOULD BE A LITTLE
MORE ATTENTION FROM MY
PERSPECTIVE AND TRYING TO
UNDERSTAND HOW TO REGULATE THE
RESILIENCE BASED ON REGULATING
AN EPIGENETIC STATE.
AND THE OTHER IS A LOT OF CELLS
ARE -- AS WE'VE HEARD A LOT
ABOUT, GO DOWN A PATHOLOGICAL
PATH, BUT WE HAVEN'T TALKED TOO
MUCH ABOUT HOW THERE'S AN
OPPORTUNITY FOR RESILIENCE
THERE.
AND IN THAT CASE, THE RESILIENCE
IS BASED ON REVERSING THE SPRING
THAT WENT -- THE SPRING THAT
SPRUNG TOO FAR IN THE FIRST
IMAGE THAT I THINK WAS
PRESENTED.
I WOULD SAY RESILIENCE IS AN
OPPORTUNITY FOR REVERSING AT A
CELLULAR LEVEL AND MOLECULAR
LEVEL SOME OF THE THINGS THAT
WE'VE LEARNED HAVE GONE WRONG
AND TO THE EXTENT THAT WE CAN
TAKE ADVANTAGE OF THE KNOWLEDGE
ABOUT HOW THINGS HAPPENED, TO
REVERSE IT, THEN THAT'S APLAYING
AAPPLYINGA NEW LEVEL OF RESILIEE
FIELD.
FROM MY PERSPECTIVE, THESE ARE
THE TWO THINGS THAT I WOULD DO
THE MOST.
>> CAN YOU RESPOND TO THAT REAL
QUICK, JUST SHORT?
I THINK THAT'S VERY RIGHT, I
THINK THERE'S SOME VERY
INTERESTING LITERATURE OUT THERE
FROM THE LUNG FIELD BUT ALSO
BEYOND ABOUT THIS EXACT SORT OF
FUNDAMENTAL CONCEPT OF EP
GENETIC STATE AND VERY COOL
BECAUSE AS YOU GET MORE AND MORE
OPEN CHROMATIN, YOUR SORT OF
ACCESSIBLE STATE SPACE BROADENS.
THE CELL STATES THAT ARE ALLOWED
BROADEN.
SO EVEN IF YOU PUT A CELL WITHIN
A VERY FOCUSED MICROENVIRONMENT
AND YOU GET REALLY GOOD CELLULAR
DIFFERENTIATION, YOU'RE GOING TO
GET A DIFFERENT -- THAT CELL HAS
A DIFFERENT ABILITY TO RESPOND
BASED ON ITS EP GENETIC STATE.
IF CHROMATIN IS ONLY ALLOWING
THOSE CERTAIN GENES, THEN IT'S
STUCK THERE, BUT IF IT HAS A
BROADER CHROMATIN ACCESSIBILITY,
THEN IT CAN GET PUSHED AROUND
MORE EASILY.
SO I THINK IT PROBABLY IS VERY
DEEPLY RELATED TO WHAT WE'RE
TALKING ABOUT AND WE SEE IT IN
AGING AS WELL, WHERE THERE'S
THIS SORT OF PROGRESSIVE
EPIGENETIC CHANGE IN A LOT OF
DIFFERENT CELL SYSTEMS.
>> I THINK WE SHOULD BE PAYING
MORE ATTENTION TO THIS.
IN TERMS OF RECOMMENDATIONS FOR
THE NHLBI, THAT WOULD BE ONE OF
MINE.
THE OTHER WOULD BE REVERSING THE
PATHOLOGICAL CELL STATE.
BOTH VERY DIFFERENT THAN TALKING
ABOUT PEOPLE.
>> WE ALSO PROBABLY NEED TO LOOK
AT EARLY DISEASE BECAUSE MOST OF
THE TIME THAT WE ARE JUST
LOOKING AT IN DISEASE JUST
BECAUSE WHERE TISSUES ARE
AVAILABLE, SO RESILIENCE IS
PROBABLY LIKE NOT WHERE WE ARE
LOOKING AT MAYBE IT'S NOT
RESIDENT ANYMORE BECAUSE THEY
HAVE COMMITTED TO GO TO THIS
DISEASE PATHWAY.
ESPECIALLY FOR THE HUMAN CELLS,
HUMAN TISSUES, BECAUSE THAT'S
WHAT IS AVAILABLE, HOW TO GET
MONORMAL, DURING THE PROCESS OF
DISEASE DEVELOPMENT.
>> I HATE TO ACTUALLY BREAK THE
LINE BECAUSE OF THE EPIGENETIC
CHANGES ARE VERY INTERESTING,
BUT SOME MINUTES AGO, WHAT CAME
UP TO MY MIND WAS SOMETHING THAT
COULD BE A POTENTIAL WAY OF
EXPLORING THE QUESTIONS THAT WAS
POSED IN TERMS OF HOW WE CAN
UNDERSTAND A LITTLE BIT MORE
ABOUT THE NEURAL.
AND COMES UP TO MY MIND ALWAYS
SOME THINK THAT IT'S KIND OF
LIKE READILY AVAILABLE.
BUT I DON'T KNOW -- WHEN I SAY
MODEL, ESSENTIALLY THE HUMANS,
PATIENTS WHO HAVE UNDERGONE LUNG
TRANSPLANTATION IN WHICH YOU
HAVE ALL OF THE CONNECTIONS
BETWEEN THE NEURONS, SO I WAS
WONDERING IF -- HAS ANYBODY
THOUGHT ABOUT WHAT ARE THE
DIFFERENCES?
THIS IS A VERY DIRTY SYSTEM TO
ACTUALLY WORK IF WE CAN SAY SO,
BECAUSE THERE IS ALL OF THE
SUPPRESSION AND ALL OF THE
THINGS THAT MAKE THINKS VERY
COMPLICATED, BUT TO MY MIND, ONE
OF THE FEW WAYS OF ADDRESSING
POTENTIAL QUESTIONS LIKE THOSE
THAT WERE RAISED.
>> I WANT TO RAISE ANOTHER
POINT, I MEAN CHALLENGES IN
TERMS OF HOW WE STUDY THIS
RESILIENCE.
ONE THING, SYSTEMS DIFFERENCES
BETWEEN MOUSE AND HUMAN, ANOTHER
IS A DOSE EFFECT RKT AND IT'S A
WHOLE DURATION THAT THE UNIVERSE
WILL GET TO -- TREATED IN CELLS
OF MICE, AND THEY USE THAT, YOU
CAN ALSO TREAT A WEEK, A MOUSE
OR WHAT KIND OF CRITERIA, THE
HALLMARKS OF RESILIENCE, SO
EXACTLY WHAT KIND OF -- I MEAN,
CURIOUS, WILL LOOK AT --
RESILIENCY THAT'S MOLECULAR
MARKER OF SOME FUNCTIONALITY OR
EXPRESSION PROFILES, AND I THINK
IT'S -- HAVE TO THINK ABOUT --
AND WHAT KIND OF CRITERIA WE
LOOK AT IS THE WHOLE EXPRESSION
FUNCTIONALITY.
>> I ACTUALLY WANT TO BRING UP
ANOTHER CRAZY -- I WAS THINKING
YOU KNOW WHEN YOU TERRITORY
CANCERS, YOU DO HAVE A
RESISTANCE IN THE LABS, AND I
DON'T WORK IN CLINICS ANYMORE,
BUT I WONDER, SAY YOU HAVE THAT
FIBROSIS, ASTHMA, CHRONIC
DISEASE, COPD, THE SYSTEM WOULD
DEVELOP A RESISTANCE BY --
DIFFERENT CELL TYPES AND HAVE
ANYBODY THINK ABOUT THAT WAY?
CAN WE USE SYSTEMATIC BIOLOGIES
TO ADDRESS THOSE ISSUES?
MAYBE CLINICIANS HAVE AN IDEA?
>> CAN YOU REPEAT THE QUESTION
AGAIN?
>> THE QUESTION WOULD BE, YOU
KNOW, WHEN YOU TREAT CANCER,
THEY DEVELOP THE SYSTEMS O RECYR
RELAPSE AFTER TREATMENT IF YOU
TREAT PATIENTS WITH FIBROSIS AND
CHRONIC DISEASE LIKE ASTHMA THE
SYSTEM NOW WILL ALSO DEVELOP A
LITTLE RESISTANCE IN SOME WAY,
MOBILIZE DIFFERENT TYPE OF CELL
TYPES.
WOULD YOU CONSIDER THIS
POSSIBILITY IN CLINICS?
>> WELL, I MEAN, THIS CAME UP --
HALL ASKED US -- WELL, NO, NOT
HAL BUT AMANDA I THINK ASKED
ABOUT WITH RESPECT TO
MESENCHYMAL STROMAL CELL-BASED
THERAPY, MULTIPLE
ADMINISTRATIONS WITH DIFFERENT
SOURCES.
CELLS OF DIFFERENT ORIGIN.
THAT ARGUABLY COULD MITIGATE ANY
POTENTIAL IMMUNE RESPONSE TO A
GIVEN EPITOPE ON ANY ONE GIVEN
SOURCE.
I DON'T KNOW HOW TO ANSWER THAT.
MY GUT WOULD BE THAT IF YOU CAN
GET STRUCTURAL REPAIR WITH
WHATEVER PROGENITOR CELL OR I
IPSC, THE HALF-LIFE IS QUITE
LONG OF TURNOVERS PRESUMABLY
LIKE THE NATIVE EPITHELIUM, THEN
YOU WOULDN'T NEED TO NECESSARILY
DO THAT.
IT WOULD TAKE YEARS AND YEARS
AND YEARS AND YEARS TO REDEVELOP
A PATHOLOGY.
SO CONCEPTUALLY INTERESTING,
CLINICALLY PROBABLY NOT
NECESSARY, NOT UNTIL WE CAN FIX
IT THE FIRST TIME AROUND AT
LEAST.
>> TO ME, A LOT OF CHRONIC LUNG
DISEASE ACTUALLY WE DON'T EVEN
HAVE TREATMENT IS I THINK IN
MEDICINE, THE REAL, LIKE, GOOD
EXAMPLE OF THE REAL TREATMENT IS
USUALLY THE ANTIBODY FOR THE
INFECTION, SO IN THAT CASE,
USUALLY YOU DEVELOP RESISTANCE
SOMETIMES BUT I DON'T THINK IN
CHRONIC LUNG DISEASE, WE DON'T
HAVE THOSE KIND OF ANTIBIOTIC
HRB LIKE THE TREATMENT FOR
PULMONARY FIBROSIS OR EMPHYSEMA,
COPD, SO I THINK THAT'S AN
INTERESTING CONCEPT BUT I DON'T
THINK WE CAN THINK ABOUT THAT
LEVEL YET AT THIS POINT.
>> I WANT TO BOUNCE THIS TO JOHN
ENGELHART.
ONE OF THE ISSUES WITH
CELL-BASED THERAPY FOR CYSTIC
FIBROSIS IS THAT IT COULD BE THE
CASE IN WHICH THE BODY AND LET'S
SAY A PATIENT WILL RECOGNIZE AN
ATTACK SEQ PROTEIN AS A FOREIGN
EPITOPE BECAUSE THEY DIDN'T GROW
UP WITH IF THE IMMUNE SYSTEM
DIDN'T DEVELOP.
I'M NOT SURE IF THE CFTR
MODULATOR DATA HAS ADDRESSED
THAT, BUT IT RAISES THE
POSSIBILITY THAT IF YOU HAVE A
GENETIC DISEASE, MISFOLD FOLDEDR
POORLY EXPRESSED OR WHATEVER
PROTEIN AND NOW YOU HAVE IT,
DOES THAT BRING IN, IN RESPONSE,
A SHORTENED HALF LIFE OF YOUR
THERAPY?
SO JOHN, ARE YOU HERE?
OR ANYONE ELSE WOULD LIKE TO
ADDRESS THAT QUESTION?
>> I'M NOT SURE IF JOHN WAS
STILL HERE, BUT I WANTED TO
CHECK WITH OUR NHLBI COLLEAGUES,
AS WE'RE UP AT 3:45 TO SEE IF WE
SHOULD MOVE ON TO THE CLOSING
REMARKS.
>> YEAH, WE CAN.
I'D ALSO LIKE TO ASK THE PANEL
ONE QUESTION.
I KNOW THIS WORKSHOP, WE KIND OF
FOCUS ON THE PROGENITOR CELLS
AND THE ROLE IN LUNG RESILIENCE,
BUT I'M SURE THERE ARE A LOT OF
CELLS IN THE LUNG AND OTHER --
CELL PROBABLY VERY IMPORTANT TOO
IN THE PRESERVED LUNG OR LOSS OF
LUNG FUNCTION TRAJECTORY, SO
JUST WANTED TO KNOW, YOU KNOW,
THE BIG PICTURE OF LUNG
RESILIENCE, WE HAVEN'T DISCUSSED
OTHER CELLS, SO THE NEURON, WE
DIDN'T REALLY DISCOVER DISCOVER
MUCH OTHER CELL TYPES.
THE CRITICAL ROLE OF OTHER CELL
TYPE IN THE LUNG RESILIENCE.
>> I THINK SAM AND I MIGHT BE
ABLE TO BE HERE MANY MORE HOURS
DISCUSSING THIS TOPIC HERE.
I MEAN, THERE'S OBVIOUSLY -- WE
DIDN'T GET INTO FIBROBLASTS OR
FIBROBLASTS SUBSETS OR WHAT
NAMES FOR THEM COULD BE.
I THINK THIS IS PROBABLY THE
TOPIC FOR ANOTHER MEETING
POTENTIALLY.
IN ADDITION TO THE REST OF THE
EPITHELIAL CELLS THAT FOLKS ARE
CATALOGING.
I KNOW THAT ON THAT TOPIC, I SAW
THAT JACKIE, I BELIEVE, HAD SENT
OUT AN EMAIL EARLIER TODAY ON
KIND OF MAPPING A LUNG CELL
ATLAS AS PART OF THE HUBMAP
PROJECT SO I'M SURE THERE WILL
BE MORE DISCUSSION ON THESE
POINTS THERE.
I THINK AT LEAST FROM WHERE --
AT LEAST SOME OF OUR LAB SITS IN
TERMS OF THE SINGLE CELL SPACE,
WE ARE VERY MUCH STILL KIND OF
IDENTIFYING AND ENUMERATING THE
DIFFERENT CELL SUBSET THAT ARE
POSSIBLE, RIGHT?
ONE OF THE THINGS THAT THEY'VE
GOTTEN TO THE BOTTOM OF
EVERYTHING, AND THEN YOU LEARN
ABOUT THE IONOCYTE, YOU LEARN
ABOUT OTHER KIND OF UNIQUE
CELLS, OUR GROUP HAS FOUND ONE
IN THE NOSE RECENTLY AND SO
FORTH.
SO AS YOU START TO BUILD OUT
THAT PARTS LIST, YOU RAPIDLY
SCALE ON THE POTENTIAL
INTERACTIONS THAT MAY BE PRESENT
ACROSS THESE SYSTEMS, NOT TO
MENTION THE IMPORTANCE OF THE
ENVIRONMENTAL LAYER ON TOP OF
THIS, AND SO I THINK THAT
CLEARLY FOOD FOR THOUGHT GOING
FORWARD, BUT IN KEEPING THE
CONCEPT OF RESILIENCE AND
RESILIENCE IS ANCHORED WITHIN
THE PLASTICITY OF THESE
PROGENITOR CELLS DEPENDING ON
HOW FIXED OR UNFIXED THEY MIGHT
BE IS CERTAINLY SOMETHING
INTERESTING TO THINK ABOUT
MOVING FORWARD.
>> THANK YOU.
IF YOU DON'T HAVE ANY OTHER
COMMENTS, YES, WE CAN CLOSE THIS
WORKSHOP.
I JUST WANTED TO SAY, THIS IS A
VERY, VERY SUCCESSFUL WORKSHOP,
I HAVE SEEN -- I HAVE
EXPERIENCED, I THINK, I
DEFINITELY MYSELF LEARNED A LOT
IN THE LAST TWO DAYS.
AGAIN, I JUST WANT TO THANK OUR
EXTRAORDINARY CO-CHAIRS,
MODERATORS, ALL THE SPEAKERS,
FOR YOUR EFFORT AND CONTRIBUTION
OF YOUR WISDOM TO THIS WORKSHOP.
I ALSO WANT TO THANK ALL THE
PARTICIPANTS TO STICK AROUND TO
THE END OF THE WORKSHOP, BY
LOOKING AT THE SCREEN TO SEE HOW
MANY PEOPLE AT THIS TIME, AND
IT'S REALLY HARD, TWO DAYS IT
WAS A WORKSHOP, SO I REALLY -- I
PROBABLY MENTIONED AT THE
BEGINNING OF THE WORKSHOPS, WE
PLAN TO PUBLISH EXECUTIVE
SUMMARY, ON THE WEBSITE
HOPEFULLY IN A FEW MONTHS.
WE ALSO PLAN TO PUBLISH A WHITE
PAPER TO SUMMARIZE THIS
WORKSHOP.
WE'RE HOPING PROBABLY WITHIN A
YEAR BUT DEFINITELY WITH YOUR
GUYS' HELP, WE HAVE SOME
DISCUSSION WITH THE CO-CHAIRS
AND WE ACTUALLY HAVE TWO
SCIENTIFIC -- IT'S TWO, DR. HAN
AND DR. -- THEY ALSO --
LEADERSHIP SO STAY TUNED.
ALSO JUST WANTED TO REMIND, THIS
WORKSHOP HA HAS BEEN VIDEOCASTED
AND THE RECORDING WILL BE
AVAILABLE, I THINK ON NIH
VIDEOCAST PAST EVENTS ONE WEEK
FROM TODAY.
SO IF YOUR COLLEAGUES,
ESPECIALLY IF YOUR TRAINEES HAVE
NOT GOT AN OPPORTUNITY TO
PARTICIPATE IN THIS WORKSHOP, IF
THEY'RE INTERESTED, I WOULD
ENCOURAGE THEM TO DO SO, TO LOOK
AT THOSE RECORDINGS A WEEK FROM
TODAY JUST BY THE TITLE, THEY
WILL BE ABLE TO FIND IT A WEEK
FROM TODAY.
WE WILL BRING ALL THE GREAT
IDEAS AND RECOMMENDATIONS FOR
THE DISCUSSION INTERNALLY.
WE HOPE SOME RECOMMENDATIONS
WILL BE IMPLEMENTED IN THE NEXT
FEW YEARS.
I THINK THAT'S WHAT I HAVE.
I'D LIKE TO BRING MYCOLEAD DR
DR. CHRISTINE CHRISTIAN GOMEZ TO
SAY A FEW WORDS.
>> I JUST WANT TO TAKE THE
OPPORTUNITY TO ALSO THANK YOU,
DR. LIU AND DR. GOMEZ FOR
LEADING THIS AND REALLY
SPEARHEADING THIS EFFORT.
NONE OF THIS WOULD HAVE BEEN
POSSIBLE WITH EITHER OF YOU, AND
ALSO TO SHAWN AND JACKIE TO
PROVIDING SUPPORT AS WELL, IT
MAKES OUR PROCESS QUITE EASY AND
LETS US FOCUS ON THE SCIENCE AND
THE DISCUSSION AND THEN
EVERYTHING HAPPENS SEAMLESSLY IN
THE BACKGROUND.
SO I JUST WANT TO MAKE SURE THAT
WE CAN ALL PUT OUR KIND OF
VIRTUAL HAND TOGETHER FOR THEIR
INCREDIBLE SUPPORT OF THIS
WORKSHOP, AND THEN CHRISTIAN, I
MIGHT HAVE HEARD YOU COME
THROUGH, SO I WILL HAND TO YOU.
>> CAN YOU HEAR ME NOW?
>> YES.
>> PERFECT.
SO I GUESS TO STAY WITHIN THE
END OF THE WORKSHOP, I JUST WANT
TO THANK EVERYONE FOR YOUR
PARTICIPATION AND CONTRIBUTIONS.
WE HAD A LOT OF GOOD INSIGHT
COME IN THESE TWO DAYS, AND WE
EXPECT THAT THIS WILL PAVE THE
WAY FOR ADVANCES THAT WILL
INCREASE OUR UNDERSTANDING OF
PROGENITOR RESILIENCE AND ALSO
CHRONIC DISEASES.
WE LOOK FORWARD TO CONTINUING
THAT DIALOGUE.
WE MAY BE SETTING THE GROUND, I
THINK, TO BE WHAT I CALL -- I
WOULD LIKE TO CALL THE PILOTS OF
LUNG HEALTH, AND HOW THIS CAN
HELP TO DEFINE A MORE RESILIENT
FUTURE FOR LUNG HEALTH IS STILL
AHEAD OF US, YOU KNOW.
SO PLEASE STAY ENGAGED AND
CONNECTED AS WE MOVE FORWARD.
AND THANK YOU ALL FOR MAKING
THIS WORKSHOP A REMARKABLE
SUCCESS, AND WITH THAT, I CLOSE.
THANK YOU SO MUCH.
>> THANK YOU.
HAVE A GREAT DAY.
A GREAT EVENING.