GOOD MORNING, I WOULD ALSO
LIKE IT SAY GOOD AFTERNOON OR
EVENING TO SOME OF OUR
PARTICIPANTS WHO MAY LIVE IN
DEFINITE PART OF THE GLOBE.
WOW, SO GOOD TO SEE YOU ALL
HERE.
WELCOME TO THE 2 DAY WORKSHOP
PROGENITOR RESILIENCE AND THE
EARLY ONSET OF CHRONIC LUNG
DISEASE HOSTED BY THE DIVISION
OF LUNG DISEASE AT THE NATIONAL
HEART, LUNG AND BLOOD INSTITUTE
OF THE NATIONAL
INSTITUTE OF
HEALTH.
ON BEHALF OF OUR DIRECTOR OF
DIVISION, DR. MATUTE-BELLO, AND
THE REST OF THE TEAM, I WOULD
LIKE TO WELCOME YOU TO TODAY'S
PRESENTATIONS.
I AM DR. LU, PROGRAM DIRECTOR
OBSTRUCTIVE LUNG DISEASE BRANCH
OF THE NHLBI, I AM HAPPY TO
REPORT THAT MORE THAN 400 PEOPLE
REGISTERED FOR THIS WORKSHOP.
I AM SURE THEY ARE NOT COMING AT
THE SAME TIMES BUT I'M SURE THEY
ARE COMING IN AND OUT, SO IN
TOTAL WE HAVE 400 REGISTRATIONS.
THANK YOU AGAIN.
THIS WORKSHOP IS ALSO BEING LIVE
STREAMED AT THE NIH VIDEOCAST
AND RECORDED.
RECORDINGS WILL BE AVAILABLE IN
THE NIH VIDEOCAST ONLINE 1 WEEK
WITHIN TODAY .
OUR ACTING DIRECTOR
DR. MATUTE-BELLOS, PLANNED TO BE
HERE, HOWEVER, HE HAS ANOTHER
COMMITMENT WHICH HE HAS --
ABSOLUTELY HAS TO ATTEND.
SO I'M SURE YOU UNDERSTAND THAT.
NOW, I WOULD LIKE TO THANK OUR
CO-CHAIRS FOR THEIR
EXTRAORDINARY LEADERSHIP IN
PLANNING THIS WORKSHOP.
I WOULD ALSO LIKE TO EXPRESS OUR
GRATITUDE TO THE DIVERSE PANEL
OF MODERATORS AND SPEAKERS FOR
YOUR INSIGHT AND THE
CONTRIBUTION TO THIS IMPORTANT
EVENT.
I WANT TO THANK OUR AUDIENCE,
PARTICULARLY THOSE WHO OVERCOME
CHALLENGING TIME ZONE
DIFFERENCES.
I WOULD ALSO LIKE TO THANK NHLBI
WORKSHOP SUPPORT TEAM FOR THEIR
ASSISTNESS, AND LAST BUT NOT
LEAST, I WOULD LIKE TO THANK OUR
LEADERSHIP, DIVISION LEADERSHIP
AND MY CO-LEAD DR. KRISTI GOMEZ,
AND OTHER COMMITTEE MEMBERS FOR
THEIR SUPPORT AND CONTRIBUTIONS.
PARTICULARLY, THE DIVISION OF
LUNG DISEASE IS COMMITTED TO
ADVANCING RESEARCH THAT NOT ONLY
ADDRESSES THE COST, DIAGNOSIS,
PREVENTION ASK TREATMENT OF LUNG
DISEASE AND SLEEP DISORDERS BUT
ALSO EXPLORED INNOVATIVE
APPROACH TO PROMOTE RESILIENCE
OF LUNG TISSUE IN ITS CELLULAR
COMPONENT AND PRESERVE LUNG
[INDISCERNIBLE].
BY UNDERSTANDING THE CURRENT
RESILIENCE OF PROGENITOR CELLS
WE UNCOVER NOVEL PATHWAYS FOR
MITIGATION OF THE EARLY ONSET
AND PROGRESSION OF CHRONIC LUNG
DISEASE .
THAT BEING SAID THE OVERARCHING
GOAL OF THIS WORKSHOP IS TO
BRING TOGETHER MULTIDISCIPLINARY
RESEARCHERS, INCLUDING PROMINENT
MAGAZINE SCIENTISTS AND STEM
CELL AND PROGENITOR SCIENTISTS
TO DISCUSS THE CURRENT STATE OF
SCIENCE AND IDENTIFY CRUCIAL
RECENT GAPS, CHALLENGES,
OPPORTUNITIES AND KEY QUESTIONS
IN THE AREA OF LUNG PROGENITOR
CELL RESILIENCE AND IMPLICATIONS
IN THE EARLY ONSET AND
PROGRESSION OF CHRONIC LUNG
DISEASE INCLUDING MECHANISMS AND
POTENTIAL PROGENITOR BASED
BIOMARKERS AND THERAPEUTIC
TARGETS.
WE BELIEVE THIS WORKSHOP IS
TIMELY, AND IS ALIGNED WITH
NHLBI'S STRATEGIC OBJECTIVES.
THANK YOU FOR YOUR
[INDISCERNIBLE].
WE LOOK FORWARD TO A SUCCESSFUL
WORKSHOP AND YOUR ACTIONABLE
RECOMMENDATIONS.
NOW, LET ME WELCOME MY CO-LEAD
DR. KRISTI GOMEZ TO SAY A FEW
WORDS AND INTRODUCE OUR
INCREDIBLE CO CHAIRS, THANK YOU.
KRISTI TAKE IT AWAY.
>> THANK YOU SO MUCH QING, THIS
IS EXCITING.
GOOD MORNING AND WELCOME TO THE
WORKSHOP, I AM CHRISTIAN GOMEZ
PROGRAM OFFICER AT THE DIVISION
OF LUNG DECS IN NHLBI.
IT'S A PLEASURE TO BE HERE WITH
YOU TODAY AS WE DELVE INTO THE
CRITICAL ISSUES SURROUNDING
CHRONIC LUNG DECS WITH A
PARTICULAR FOCUS ON ENHANCING
RESILIENCE IN PROGENITOR CELLS.
I AM HONORED TO INTRODUCE OUR
CO-CHAIRS FOR THIS WORKSHOP WHO
WILL BRING EXCEPTIONAL EXPERTISE
AND INSIGHT INTO THIS VITAL AREA
OF RESEARCH.
FIRST WE HAVE DR. RAVI KALHAN,
WITH THE NORTHWESTERN UNIVERSITY
FINE BERG SCHOOL OF MEDICINE,
DR. KALHAN IS FOCUSING ON
EPIDEMIOLOGY AND MANAGEMENT OF
THESE CONDITIONS, HIS WORK
EMPHASIZES EARLY DETECTION AND
INTERVENTION WITH AN INTEREST IN
HOW ENHANCING THE RESILIENCE OF
LUNG TISSUES CAN ALTER THE
DISEASE TRAJECTORY.
HIS CONTRIBUTIONS HAVE BEEN
INSTRUMENTAL IN ADVANCING OUR
UNDERSTANDING OF HOW TO FORTIFY
THE BODY'S NATURAL DEFENSES
AGAINST CHRONIC LUNG DISEASES.
JOIN DR. KALHAN, AND
DR. [INDISCERNIBLE] A RENOUNCED
RESEARCHER IN IMMUNOLOGY AND
RESPIRATORY DISEASES.
HE IS AT THE FOREFRONT OF
EXPLORING CELLULAR AND MOLECULAR
MECHANISMS THAT CONTRIBUTE TO
CHRONIC LUNG DECS.
HIS RESEARCH DELVES INTO THE
RESILIENCE OF THE NEW SYSTEM AND
IN THE DIRECTION WITH LUNG
PROGENITOR CELLS, AIMING TO
DISCOVER NOVEL THERAPEUTIC
TARGETS BY APPLYING
[INDISCERNIBLE] TECHNIQUES SUCH
AS [INDISCERNIBLE], SEEKS TO
STRENGTHENS THE RESILIENCE OF
LUNG TISSUE, ULTIMATELY
IMPROVING OUTCOMES FOR PATIENTS
WITH LUNG DECS.
TOGETHER OUR CO-CHAIRS BRING A
WEALTH OF EXPERTISE AND A SHARED
VISION ON ENHANCING RESILIENCE
AS A NEEDS TO COMBAT THE EARLY
ONSET OF CHRONIC LUNG DISEASES.
THEIR LEADERSHIP WILL GUIDE US
TOGETHER TOWARDS MINIMAL
DISCUSSIONS, MEANINGFUL
DISCUSSIONS AND SOLUTIONS.
THANKS, YOU ALL FOR JOINING US
TODAY.
I LOOK FORWARD TO THE
COLLABORATIVE EFFORTS AND
VALUABLE EXCHANGES THAT WILL
TAKE PLACE DURING THE WORKSHOP
AS WE WORK TOGETHER TO BUILD A
MORE RESILIENT FUTURE FOR LUNG
HEALTH.
RAVI AND JOSE, TAKE IT AWAY.
>> THANK YOU VERY MUCH
DR. GOMEZ, AND DR. LU FOR THE
NICE INTRODUCTION.
I WILL KEEP MY OPENING REMARKS
BRIEF, JUST TO HELP INTRODUCE
RAVI HERE TO SET THE STAGE.
I ACTUALLY FIRST MET RAVI IN THE
PLANNING OF THIS SYMPOSIUM AND I
WAS WONDERING, YOU KNOW KIND OF
HOW CHRISTIAN AND QING HAD
THOUGHT TO BRING US TOGETHER.
AND IN THE EVOLUTION OF THE
PLANNING FOR THIS, IT BECAME
CLEAR THERE'S A VERY INTERESTING
GAP TO EXPLORE BETWEEN TAKING AN
EPIDEMIOLOGICAL AND POPULATION
HEALTH VIEW ON THESE ISSUES OF
CHRONIC LUNG DISEASES AND TYING
IT TO THE EMERGENT WORK ON OUR
UNDERSTANDING OF LUNG PROGENITOR
CELLS AND HOW THEY CAN HELP US
TO UNDERSTAND AND HOPEFULLY
BUILD A MORE RESILIENT LUNG
ECOSYSTEM.
NOW, THE WORD RESILIENT MAY BE
NEW FOR ME, MAYBE NEW FOR OTHERS
IN THE AUDIENCE AND I THINK THE
IDEA OF RESILIENCE IS SOMETHING
WE WILL HAVE TO CAREFULLY
EXPLORE OVER THE NEXT COUPLE OF
DAYS, BOTH IN TERMS OF HOW WE
ALL USE THE WOORDER AND HOW WE
DEFINE IT AND THE MOLECULAR AND
CELLULAR AND SCALING BACK OUT TO
WHAT IT ACTUALLY MEANS TO NOT
ONLY REBUILD THE LUNGS BUT ALSO
RESILIENT COMMUNITY WHEN IS IT
COMES TO LUNG HEALTH.
SO I CAN GIVE A FEW INTRODUCTORY
NOTES ON RAVI.
SO RAVI IS CURRENTLY THE LOUIS
SISM SON, AT THE FINESCHOOL
MESSAGE AND FOCUSED ON LIFE
COURSE TRANSITIONS FROM
RESPIRATORY HEALTH TO CHRONIC
LUNG DISEASE, OF COURSE
SOMETHING VERY CENTRAL FOR THIS
WORKSHOP HERE, AND TO HELP US --
I WILL HAND IT OVER TO DR. RAVI
KALHAN TO GIVE THE BRIEF KEY
NOTE HERE TO SET THE STAGE FOR
THE WORKSHOP.
SO PLEASE, RAVI.
>> THANKS SO MUCH JOSE AND
CHRISTIAN AND QING AND GIVING ME
THE OPPORTUNITY TO CO-CHAIR AND
GIVING OPENING FRAMEWORK AROUND
HOW WE CAN THINK ABOUT
RESILIENCE IN THE CONTEXT OF
CHRONIC LUNG DISEASE.
THE NEXT SLIDE WOULD BE GREAT.
SO THIS IS A LITTLE BIT OF A
TALL ORDER TO DEFINE RESILIENCE
WHICH I THINK IS A CRITICAL STEP
TO PROMOTE RESPIRATORY HEALTH.
I KIND OF FEEL LIKE A CLINICAL
EPIDEMIOLOGIST AND A SEA OF
DISCOVERY SCIENTISTS SO I WILL
ASPIRE TO GIVE YOU A FRAMEWORK
FOR WHY WE NEED TO THINK ABOUT
THIS CLINICALLY, HOW WE CAN
DEFINE RESPIRATORY HEALTH AND
THEN SHIFT OUR 2-DAY
CONVERSATION TOWARD RESILIENCE.
NEXT SLIDE.
SO WHEN WE DEFINE RESILIENCE, IT
SEEMS TO ME THIS TAKES A FEW
STEPS AND WE'RE GOING TO TRY TO
ACCELERATE SOME OF THESE STEPS
QUICKLY.
SO 1 IS HOW DO WE ACTUALLY
DEFINE RESPIRATORY HEALTH.
WE NEED TO ASK THE QUESTION
WHETHER IT'S POSSIBLE TO PROMOTE
RESPIRATORY HEALTH.
IN MY TIME AS A PULMONARY
POSITION, AND IN NIGH TRAINING
WE'VE OFTEN THOUGHT ABOUT
RESPIRATORY HEALTH AS SOMETHING
THAT ONLY GETS WORSE ACROSS THE
LIFE COURSE.
PEOPLE KIND OF PEAK 1 FUNCTION
AND THEN DECLINE.
I'M GOING TO TRY TO HELP
UNDERSTAND WHAT RESPIRATORY
INVESTIGATORS THINK ABOUT
RESILIENCE TO DATE.
THE ANSWER IS THERE HASN'T BEEN
THAT MUCH THOUGHT FROM THE
CLINICAL COMMUNITY ABOUT
RESILIENCE AT LEAST, AND THEN
ASK THE QUESTION, AND MAYBE SHOW
A LITTLE BIT OF PRELIMINARY
STUFF FROM OUR OWN WORK ABOUT
HOW WE CAN ULTIMATELY DEVELOP
BIOLOGIC DEFINITIONS AND TARGETS
THAT DISTINGUISH SUSCEPTIBILITY
FROM RESILIENCE.
NEXT SLIDE.
SO IF WE THINK ABOUT HOW LUNG
FUNCTION AS BEEN CONCEPTUALIZED
ACROSS PEOPLE'S LIFE COURSE,
WE'VE OFTEN THOUGHT ABOUT
TRAJECTORIES, PEOPLE DEVELOP
THROUGH CHILDHOOD AND THEY REACH
PEAK 1 FUNCTION, THAT'S OFTEN
THOUGHT TO BE IN YOUNG
ADULTHOOD, SOME PEOPLE REACH
HIGHER PEAKS THAN OTHERS, THAT'S
BEEN A POINT OF EMPHASIS IN THE
COPD COMMUNITY LATELY THAT THERE
MIGHT BE PEOPLE WHO HAVE
RESILIENCE BASED ON BETTER LUNG
DEVELOPMENT AND BETTER LUNG
GROWTH THAN OTHERS WHO HAVE LESS
PEAK LUNG FUNCTION.
IT'S BEEN TAUGHT THAT AFTER
MAXIMUM LUNG FUNCTION IS
ACHIEVED WE HAVE AN UNRELENTING
DECLINE IN LUNG FUNCTION, THINGS
ONLY GET WORSE ACCORDING TO THE
DECLINING FEATURES OF
RESPIRATORY HEALTH, BUT THERE
ARE SOME WHO PRESERVE IT MORE
THAN OTHERS, SOME HAVE A DECLINE
THAT DESCENDS TOWARDS DISEASE
ALATHE [INDISCERNIBLE] DISEASE
IN LUNG FUNCTION.
I THINK WE NEED TO SPEND A
LITTLE MORE TIME THINKING ABOUT
THE INTERMEDIATE PHENOTYPE OF
IMPAIRED RESPIRATORY HEALTH.
THAT IS THOSE PEOPLE SEEM TO BE
TILTING IN YELLOW HERE AWAY FROM
IDEAL RESPIRATORY HEALTH, WHAT'S
THE BEST THEY'RE GOING TO BE IN
THEIR LIFE AND WHETHER WE CAN
ACTUALLY INTERSECT THOSE PEOPLE
BY PROMOTING RESILIENCE WHETHER
WE CAN EXPERIENCE IT OR THOSE
WHO EXPERIENCE LUNG IT BUT THEY
DEVIATE ABOUT YOU WE CAN START
TO RESTORE LUNG FUNCTION.
I AM A COPD DOCTOR, I THINK WE
SHOULD DELIVER THE BEST
TREATMENTS TO PEOPLE LIVING WITH
COPD.
I THINK IT WOULD BE BETTER IF WE
COULD INTERCEPT THEIR
UNFAVORABLE TRAJECTORYINGS
BEFORE THEY DESCENDED TOWARD THE
DEVELOPMENT OF EMPHYSEMA AND
THAT'S SORT OF A KEY PARADIGM IN
PROMOTING RESPIRATORY HEALTH.
NEXT SLIDE, PLEASE.
SO WHO'S DONE THIS BEFORE, WELL
CARDIOVASCULAR PEOPLE DO THIS
ALL THE TIME AND THEY TALK A LOT
ABOUT IDEAL CARDIOVASCULAR
HEALTH THROUGH A BALANCE OF
HEALTH PROMOTION, WITH DEC
INTERCEPTION AND BY HEALTH
PROMOTION, THEY THINK ABOUT
BEHAVIORS QUIT SMOKING, BE
ACTIVE, EAT HEALTHY, LOSE SOME
WEIGHT AND THEN THEY THINK ABOUT
HOW TO INTERCEPT DISEASE FOR
THOSE WHO WHO HAVE INDICATORS OF
AWN UNFAVORABLE TRAJECTORY, ALL
OF WHICH HAVE BEEN INFORMED BY
DISCOVERY DEC THROUGH, YOU KNOW
A CENTURY NOW.
CONTROLLING CHOLESTEROL,
MANAGING BLOOD PRESSURE, AND
REDUCING BLOOD SUGAR BECAUSE
THESE ARE THINGS ON THE CAUSAL
PATHWAY TOWARDS DISEASE THAT CAN
ACTUALLY BE INTERCEPTED.
I WILL HAZARD TO SAY WE CAN
PROMOTE HEALTH IN LUNG DEC BY
TELLING PEOPLE TO STOP SMOKING,
WE HAVE DEFICIENT INFORMATION IN
EPIDEMIOLOGIC STUDIESAs WELL
AS DISCOVERY PROGRAMS TO SOME
EXTENT ABOUT ALL THE OTHER
THINGS WE CAN DO TO TRY TO
PROMOTE RES PERATTORY HEALTH
AND DWEPT DEVIATIONS FROM IDEAL
RESPIRATORY HEALTH.
NEXT SLIDE.
SO HOW CAN WE THINK ABOUT TARGET
POPULATIONS IN WHICH WE CAN
PROMOTE REZONING PIRATTORY
HEALTH?
WELL THIS WOIS WORK THAT COMES
FROM OUR GROUP WHERE WE USE A
LARGE EPIDEMIOLOGICAL TODAY
CALLED CARDIA, AN NHLBI
SPONSORED COHORT THAT RECRUIT
THE 18-30 YEAR-OLDS IN 1985 AND
FOLLOWED THEM FOR NEARLY 40
YEARS.
WE ASKED THE QUESTION WHETHER WE
COULD IDENTIFY SUSCEPTIBLE
POPULATIONS, JUST BASED ON A
SIMPLE THING, LIKE THE PRESENCE
OF RESPIRATORY SYMPTOMS WHEN
THEY ARE BUT MEAN AGE 25-27
YEARS OLD.
SO DO YOUNG ADULTS WITH
RESPIRATORY SYMPTOMS DEMONSTRATE
RISK FOR FUTURE DISEASE BECAUSE
WE NEED TO IDENT PIE AND DEFINE
LIKE I SAID BEFORE, WHAT IS
IMPAIRED VERSUS IDEAL
RESPIRATORY HEALTH BECAUSE IF WE
WILL PROMOTE RESILIENCE, WE NEED
TO UNDERSTAND THOSE DIFFERENCES,
RIGHT?
AND IT TURNED OUT THAT
INDIVIDUALS WHO REPORTED
RESPIRATORY SYMPTOMS AT MEAN AGE
25 WERE AT INCREASED RISK WHEN
FOLLOWED FORWARD FOR THE NEXT
20-30 YEARS.
INCREASED RISK FOR OBSTRUCTIVE
LUNG PHYSIOLOGY AND IN THE CASE
OF PEOPLE WHO REPORTED COUGH AND
PHLEGM AT THAT AGE, WERE AT
INCREASED RISK, 1.46 WAS THE
ODDS RATIO, 1.5 FOR THE RATIOS
OF BRONCHITIS OF HAVING CT
EMSEEMA 25 YEARS LATER.
SO THIS INDICATES TO ME THIS IS
A SUSCEPTIBLE POPULATION WHO
MIGHT COMPRISE PART OF A
DEFINITION OF IMPAIRED
RESPIRATORY HEALTH WHO SHOULD BE
TARGETED FOR HEALTH PROMOTION.
NEXT SLIDE.
WHO ELSE IS AT RISK FOR RESPIRAT
ORY DISEASE WHO ARE TARGETED
POPULATIONS?
WELL THOSE IN DISADVANTAGE
NEIGHBORHOODS, WE LOOKEDDA THE
PEOPLE MEAN AGE 28-40, AND
FOLLOWED THEM 15-20 YEARS
FORWARD AND PEOPLE WHO LIVED IN
DISADVANTAGED NEIGHBORHOODS,
INDEPENDENT OF BOTH THEIR
INDIVIDUAL INCOME, AND THEIR
CUMULATIVE PACK THEY'RE SMOKING
WERE INDEED AT GREATER RISK FOR
FUTURE DEVELOPMENT OF EMPHYSEMA
AS WELL AS FUTURE DEVELOPMENT
WITH SPIROMETRY, ANOTHER GROUP
WHO HAVE AT RISK FOR FUTURE
DISEASE AND MAYBE IMPORTANT
PEOPLE TO TARGET FOR RESPIRATORY
HEALTH PROMOTION AND
INTERVENTIONS THAT FOSTER
RESILIENCE.
NEXT SLIDE.
NOW OF COURSE NEIGHBORHOOD
DISADVANTAGE IS AN INTEGRATIVE
MEASURE RIGHT, OF RISK?
IT INVOLVES INCREASED EXPOSURE
TO BIOMASS FUEL, RISK FACTORS,
ASINERATES ARE HIGHER IN --
ASTHMA RATES ARE HIGHER IN
INDIVIDUAL NEIGHBORHOODS.
INDIVIDUALS BORN PREMATURELY MAY
EXPERIENCE THIS, EARLY LIFE MAY
IMPACT THIS, IMPAIRED GROWTH MAY
EXPERIENCE THIS, AND WE NEED TO
DIVE DEEPER IN HOW THESE FOSTER
DEVIATIONS FROM RESPIRATORY
HEALTH.
BUT IF WE THINK ABOUT
URBANIZATION GLOBALLY, THE
SOCIOECONOMIC DIVIDES THAT
EMERGE, THERE'S A TREMENDOUS
POPULATION WHO WILL REQUIRE OUR
THINK BEING HOW TO INTERVENE
UPON AND PROMOTE RESPIRATORY
HEALTH.
NEXT SLIDE, PLEASE.
SO I WAS TALKING ABOUT BEFORE
HOW WE TAUGHT OUR MEDICAL
STUDENTS AND TRAINEES FOR A LONG
TIME THERE THERE'S THIS
UNRELENTING DECLINE IN LUNG
FUNCTION AND WHEN I WAS SORT OF
EARLIER ON IN MY CAREER, I WAS
LIKE, MAN, IF THAT'S THOUGHT TO
BE TRUE, WE'RE IN BIG TROUBLE
BECAUSE IF YOU CAN'T PROMOTE
RESPAIRATTORY HEALTH SORT OF ON
THE FOUNDATIONAL IDEA THAT
PEOPLE AREN'T GOING TO BUY-INTO
THIS CONCEPT, SO WE ASKED A
QUESTION IN CARDIA, WE ASK
WHETHER PEOPLE WHO PRESERVE
PHYSICAL FITNESS ACTUALLY HAVE
RESILIENCE, IE THEY EXPERIENCE
LESS LUNG FUNCTION DECLINE AND
IT TURNED OUT THEY DID.
IT TURNS OUT, FITNESS EXPERIENCE
LESS LUNG FUNCTION DECLINE
INDEPENDENT OF WHETHER THEY
SMOKE, INDEPENDENT OF WHETHER
THEY GAIN WEIGHT, INDEPENDENT OF
WHERE THEY LIVE AND A VARIETY OF
OTHER CO VARIANTS.
SO INDEED, PERHAPS WE DO HAVE
STRATEGIES TO PROMOTE RESILIENCE
AND WE KNOW THAT EXERCISE AND
FITNESS HAVE BENEFITS,
ANTIINFLAMMATORY BENEFITS OR
THESE GLOBALLY STATED BENEFITS,
PERHAPS WE NEED TO THINK DEEPER
ABOUT HOW TO UNDERSTAND THESE
BENEFITS MORE SO IN THE LUNG,
BUT REALLY I SAY THAT THIS IS A
PARADIGM THAT THERE ARE THINGS
THAT OCCUR THAT DO ATTENUATE
THIS UNRELENTING LUNG FUNCTION
DECLINE AND MAY BE ABLE TO
PROMOTE RESPIRATORY HEALTH.
NEXT SLIDE.
SO IF I WERE TO SUM THIS UP AND
THEN MOVE A LITTLE BIT FORWARD
TO THINKING, A LITTLE BIT MORE
ABOUT RESILIENCE AND
SUSCEPTIBILITY, WE HAVE TO
DEFINE AND UNDERSTAND AN
INTERMEDIATE PHENOTYPE OF
IMPAIRED RESPIRATORY HELT.
ONE WAY IS TO THINK ABOUT LUNG
FUNCTION, LUNG FUNCTION IS
THOUGHT TO BE THE MOST WIDELY
RECOGNIZED INDICATOR OF IDEAL,
AKA RESILIENT, AKA IMPAIRED, AKA
SUSCEPTIBLE RESPIRATORY HEALTH
BUT IT'S MEASUREMENT OUTSIDE OF
THE RESPIRATORY DISEASE IS
DIFFICULT.
WE DO THESE STUDIES OVER AND
OVER AGAIN BUT NO 1 DOES THIS IN
THE CLINICAL POSITIONS OF SINGLE
RISK, AND SINGLE MEASUREMENT OF
SO, IT DOESN'T TELL YOU WHETHER
THEY TARTED LOW, HIGH,
EXPERIENCING ACCELERATED
DECLINE.
SO WE REALLY DO NEED BIOMARKERS
OF IMPAIRED RESPIRATORY HEALTH
TO ALLOW FOR MORE EFFICIENT
IDENT IIVES OF AT-RISK
INDIVIDUALS, PROVIDE MECHANISTIC
INSIGHTS INTO SUSCEPTIBILITY
VERVERSUS RESILIENCE, AND WHAT
WE CALL LUNG HEALTH ENDOTYPES
AND PROVIDE A FRAMEWORK FOR THE
INTERCEPTION OF CHRONIC LUNG
DISEASE BEFORE IT'S CLINICALLY
APPARENT.
NEXT SLIDE.
SO WHEN WE ASK CLINICAL
INVESTIGATORS WHAT A RESILIENT
PERSON IS, WE DON'T HAVE THAT
MUCH INFORMATION BUT THERE IS AN
IMPORTANT PAPER FROM ANITA O IN
SPIROMICS WHERE THEY DEVELOPED
IT YEARS AGO.
WHERE THEY DEVELOPED A CONSENSUS
DEFINITION RECARDING WHAT A
RESILIENT SMOKER IS USING A
MODIFIED DELPH I,A PROACH AND
THEY CAME UP WITH THESE
PARAMETERS.
RESILIENCE IS NOT HAVING COUGH
AND SPUTUM PRODUCTION, SHORTNESS
OF BREATH, LINES UP WITH WHAT WE
REPORTED IN CARDIA, NOT HAVING
RESPIRATORY EXACERBATIONS THINGS
THAT REQUIRED HEALTHCARE FOR
CARTIC O STEROIDS OR
ANTIBIOTICS, NOT HAVING
EMPHYSEMA ON A CT SCAN AND NOT
HAVING AIR TRAPPING ON A CT SCAN
AND THEN NOT HAVING A HIGH
ANNUAL RATE OF DECLEAN IN 1
FUNCTION.
SO IT'S THE ABSENCE OF A WHOLE
LOT OF THINGS.
THAT'S WHAT THEY USED AS THE
DEFINITION FOR RESILIENCE.
AND OF 892 SPIROMICS
PARTICIPANTS WHO HAD NORMAL LUNG
FUNCTION ONLY WOB 49
QUALIFIEDAs RESILIENT AND
AMONG HE'S PEOPLE THINKING
BIOLOGICALLY, THEY DID HAVE
LOWER LEVELS OF THESE
NONSPECIFIC INFLAMMATORY
MARKERS, TNH ALPHA FACTOR 1A AND
THEY WERE LOWER IN RESILIENT
VERSE US FAUN RESILIENT SMOKERS.
NEXT SLIDE.
I THINK WE NEED TO GET DEEPER ON
THIS AND PERHAPS NOT ONLY
KNOWLEDGE ABOUT THE ABSENCE OF
THINGS LIKE EMPHYSEMA AND AIR
TRAPPING, BUT THINK A LITTLE
DEEPER ABOUT HOW WE CAN USE
TECHNOLOGY TO UNDERSTAND EARLIER
THINGS GOING ON IN THE LUNG AND
CLINICAL POPULATION, SO 1
POTENTIAL MAY BE TO USE CT
IMAGING AND THIS IS JUST AN
EXAMPLE OF WHAT OUR
COLLABORATORS AT BRIGHAM AND
UNIVERSITY OF WASHINGTON HAVE
DEVELOPED BUT THEY HAVE THE
ABILITY TO MAKE AN ENTIRE MAP
USING A VARIETY OF FEATURES,
SOME THAT ARE VISIBLE AND SOME
ARE ATTENUATION BASED IN AN
INTEGRATED WAY USING MACHINE
LEARNING.
AND THEY CAN E DENTIFY
INTERESTING FEATURES.
ONE OF THE FEATURES WE REPORTED
ON A BIT, I DON'T HAVE TIME TO
SHARE IT WITH YOU AND NORMAL ARK
PEERING AREAS OF IATTENUATION.
MAYBE WHAT COULD BE INDICATORS
OF THE EARLIEST POINTS OF LUNG
INJURY WERE WORKING HARD TO
UNDERSTAND HOW THOSE POINTS
EVOLVE OVER TIME USING CEREAL
IMAGING AND CARDIA ACTUALLY TO
SEE WHETHER THESE POINTS OF LUNG
INJURY EVOLVE INTO VISUAL CHANGE
OR WHETHER THEY RESOLVE.
AND IF THEY DO RESOLVE, IT GIVES
US A BETTER WAI TO THINK ABOUT
WHO'S REWRILIENT VERSUS
SUSCEPTIBLE.
NEXT SLIDE.
AT THE END OF THE DAY, THOUGH, I
THINK WE'RE GOING TO HAVE TO FOR
CLINICAL POPULATIONS USE
BIOMARKERS, AND THINK ABOUT HOW
TO DEFINE SUSCEPTIBILITY VERSUS
RESILIENCE BIOLOGICALLY.
SO WE STARTED TO DO THIS, AS
PAPERS AND PRESS GABBY, AND
FELLOW AND JUNIOR FACULTY MEMBER
WITH ME AT UC DAVIS DID THIS
WORK WHERE WE ACTUALLY TOOK
ADVANTAGE OF THE LIFE COURSE
TRAJECTORIES OF LUNG FUNCTION IN
CARDIA, TO DEFINE THOSE
EXPERIENCING LUNG FUNCTION
DECLEAN AND THOSE WHO HAD
RESILIENCE, IDEAL SUSTAINED
RESPIRATORY HEALTH.
WE TOOK THESE GROUPS OF PEOPLE
AND WE CAN PROTEOMICS USING THE
SOMA [INDISCERNIBLE], WHETHER WE
COULD TRAIN A PROTEOMIC SCORE ON
SUSCEPTIBLE VERSUS RESILIENCE
AND INDEED, A NUMBER OF PROTEINS
WERE FOUND TO BE ASSOCIATED WITH
SUSCEPTIBILITY VERSUS
RESILIENCE.
WE THEN TOOK THIS PROTEOMIC
SCORE, NEXT SLIDE, AND TESTED IT
IN BOTH UK BIOBANK AND THE COPD
GENE STUDY AND DOCUMENTED THAT
AN INCREASED PROTEOMIC
SUSCEPTIBILITY SCORE WAS
ASSOCIATED WITH FUTURE POOR
RESPIRATORY OUTCOMES, INCIDENCE
IN THE UK BIOBANK, EXACERBATION
AND RISK IN THE COPD GENE, NEXT
SLIDE, AS WELL AS MORTALITY IN
BOTH OF THOSE STUDIES.
SOPHISTICATEDY WE THINK THIS IS
A REALLY PRELIMINARY STEP
FORWARD, TO THINK ABOUT HOW WE
CAN FIND WHAT WE MIGHT THINK OF
AS A CHOLESTEROL LIKE INDICATOR
FOR THE LUNG.
THOSE, A BIOMARKER THAT'S ON
CAUSAL PATHWAY THAT THINKS ABOUT
SUSCEPTIBILITY TO LUNG INJURY
AND FUTURE DISEASE AND THE
ABSENCE OF SUSCEPTIBILITY, I. E.
RESILIENCE.
WE NEED TO DO MUCH MORE WORK TO
UNDERSTAND WHETHER THOSE
BIOMARKERS, WHETHER FROM THE
PROTEOMIC DORPHY PLATFORM ARE
EXPRESSED IN LUNG TISSUE, HOW
THEY RELITE TO SORT OF THE
CAUSAL PATHWAY TOWARDS DISEASE,
WHETHER THE ABSENCE IN THE LUNG
TISSUE IS ACTUALLY INDICATIVE OF
RESILIENCE AND ALL SORTS OF
OTHER THINGS BUT THIS IS AN
INTERESTING FRAMEWORK WE THINK
TO TAKE POPULATION DORPHY AND
APPLY IT TO BIOLOGIC DORPHY
SCIENCE.
NEXT SLIDE.
SO THAT KIND OF WRAPS UP WHAT I
WANT TO SAY IN SETTING THE
FRAMEWORK BUT I WILL LEAVE YOU
WITH A FEW QUESTIONS.
WE NEED TO KNOW WHAT THE TARGET
POPULATIONS FOR US TO STUDY
SUSCEPTIBILITY VERSUS RESILIENCE
ON ARE AND TO THINK REALLY ABOUT
HEALTH PROMOTION INTERVENTIONS
THAT FOSTER RESILIENCE.
WE NEED TO UNDERSTAND WHY AND
HOW THEY WORK AND WE NEED TO USE
THE POWER OF DISCOVERY SCIENCE
TO UNDERSTAND THOSE THINGS.
WE NEED TO HAPPENED AND FIGURE
OUT WHETHER WE CAN FIND TARGETS
FOR INTERCEPTION THAT PROMOTE
RESILIENCE AND HOW WE CAN USE
DISCOVERIES REGARDING THESE
CLINICAL ENDOTYPES TO INFORM OUR
DISCOVERY SCIENCE.
WE NEED TO UNDERSTAND WHICH
TRANSLATIONAL RESEARCH PLATFOR
THE PURPOSES ARE REQUIRED FOR US
TO APPLY FUNDAMENTAL DISCOVERIES
REGARDING PROGENITOR RESILIENCE
TO HUMAN POPULATIONS AND THEN I
WANT TO THINK ABOUT WHETHER
RESILIENCE IS INDEED THE INVERSE
OF SUSCEPTIBILITY.
AND IF IT'S NOT, HOW CAN WE USE
OUR MODEL SYSTEM TO ACTUALLY
UNDERSTAND RESPIRATOR HEALTH
INSTEAD OF ALWAYS FOCUSING ON
THE DEVELOPMENT OF DISEASE?
I THINK I HAVE 1 MORE SLIDE JUST
TO THANK A BUNCH OF PEOPLE, BUT
I WILL LEAVE IT THERE.
AND I AM HAPPY TO MOVE FORWARD
WITH THE WORKSHOP.
IF THERE ARE QUESTIONS, I THINK
WE WITH PUT THEM IN THE CHAT AND
WEAVE THEM THROUGH OUR
DISCUSSION AS TIME IMZ -- GOES.
THANKS SO MUCH FOR YOUR TIME AND
ATTENTION, WE LOOK FORWARD TO A
GREAT WORKSHOP, THANKS AGAIN TO
NHLBI ORGANIZERS AND THANKS TO
JOSE FOR GRACIOUSLY TOLERATING
ME AS A CO-CHAIR.
WE CAN MOVE FORWARD I THINK.
>> OKAY, I THINK WE SHOULD START
BY THANKING RAVI AND JOSE FOR
THE STARTING SPEECH.
NOW WE MOVE TO SESSION 1
TOGETHER WITH [INDISCERNIBLE]
HERE, WE CO WHICH HAVE CHAIRED
THE FIRST SESSION, LONG STEM
PROGENITOR CELL AND THE
MECHANISM OF STRESS RESPONSE
EMPLOY SO SOUR FIRST SPEAKER
DR. SEUNG HYE HAN, ARE YOU HERE?
>> YES, I AM HERE.
>> OH, HERE.
SHE'S ASSISTANT PROFESSOR FROM
NORTHWESTERN UNIVERSITY AND
SHE'S A PHYSICIAN SCIENTISTS.
HER PRIMARY INTEREST IN THE LAB
STUDYING THE MECHANISM
EPITHELIAL REPAIR AND THE REPAIR
OCCUR RESULLING IN FIBROSIS, SO
WITHOUT FURTHER ADO, SEUNG
HYE THE STAGE IS YOURS, THANK
YOU.
>> OKAY, THIS IS ME AND THEN WE
CAN GO TO THE NEXT SLIDE I
THINK.
YES, SO HI, EVERYONE, WELCOME TO
THE WORKSHOP AND WE'RE REALLY
EXCITED TO HEAR MORE ABOUT ALL
THE GREAT TALKS SCHEDULED FOR
THIS WORKSHOP.
YOU JUST HEARD FROM DR. KALHAN
ABOUT THE LUNG HEALTH AND
RESILIENCE FROM THE POPULATION
STUDY PERSPECTIVE.
NOW WE'RE GOING TO SHIFT GEARS A
LITTLE BIT AND FOCUS ON THE MORE
FUNDAMENTAL SCIENCE, MORE
SMALLER VIEW, LUNG STEM AND
PROGENITOR CELL RESILIENCE.
NEXT SLIDE, PLEASE.
SO THE LUNG EPITHELIUM AND
SUSCEPTIBLE TO VIRAL BACTERIAL
AND TOXIC INSULT ACCIDENT, BUT
THIS SUSCEPTIBILITY IS OFFSET BY
A REMARKABLE CAPACITY TO REPAIR
AND GAIN FUNCTION AFTER INJURY
BY STEM PROGENITOR CELLS.
THIS REGENERATE CAPACITY WITH
THE LUNG WAS SEEN RECENTLY IN
MANY PATIENT RECOVERING FROM
SARS-COV-2 IN THE
[INDISCERNIBLE] ANATOMICALLY THE
LONGEST DIVIDED FUNCTIONAL THESE
THINGS BUT CONTINUOUSLY CONNECT
THE REGION.
SO I STOLE SOME PICTURES FROM
DR. MARIA BASIL'S REVIEW PAPER
AND AS YOU CAN SEE IT IS DIVIDED
PROXIMALLY, IN THE BRONCH I AND
THE SMALL AIR WAYS USUAL
BRONCHIAL AREA IN THE
RESPIRATORY AIR WAY AND THE
ALVEOLAR, AND THOSE AREAS HAVE A
DISTINCT EPITHELIAL CELL
POPULATION AND SLIGHTLY
DIFFERENT STEM PROGENITOR STEM
CELL POPULATION.
AND YOU SEE ON THE RIGHT SIDE,
YOU SEE A BUNCH OF VERY MANY
CELL POPULATIONS THAT IS
DISTINCTLY DISTRIBUTED
THROUGHOUT THE AIRWAY.
LATER IN THIS SESSION YOU WILL
HEAR FROM DR. BASIL AND
DR. [INDISCERNIBLE], FOR THOSE,
AND ON MY TALK I WILL FOCUS ON
THE WORK IN OUR REGION AND THE
FACTORS THAT CONTROL EPITHELIAL
STEM CELL FATE WHICH MAY BE A
TARGET TO PROMOTE REWRILIENCE.
NEXT SLIDE.
SO THE ALVEOLAR EPITHELIUM
CONSISTS OF TYPE 1 AND TYPE 2
CELLS.
MANY, MANY STUDY VS SHOWN THAT
TYPE 2 CELLS ARE PARTIA WILY
COMMITTED STEM CELLS THAT CAN
SELF-RENEW AND THEN GIVE RISE TO
TYPE 1 CELLS WHICH IS
RESPONSIBLE FOR THE GAS
EXCHANGE.
SEVERAL SIGNALING PATHWAYS SUCH
AS WNT, AND BETA KACCT TIENIN,
CDC 42, BMP, NOTCH, YAP/TAZ,
HAVE BEEN SUGGESTED TO CONTROL
THIS ATP STEM CELL BEHAVIOR.
NEXT SLIDE.
THANKS TO THE RECENT ADVANCES IN
SINGLE CELL SEQUENCING AND
TECHNOLOGIES, MANY GROUPS
INCLUDING OURS HAVE IDENTIFIED
AND CONFIRMED THE EXISTENCE OF
INTERMEDIATE TRANSITIONAL CELLS
BETWEEN [INDISCERNIBLE] AND
[INDISCERNIBLE] CELL
CHARACTERIZATION.
KERATIN 8 IN MICE, AND CDKN1A
WHICH IS GROWING.
IN THE PATIENT LUNGS WITH IDIO
PATHIC PULMONARY FIBROSIS AND
COVID-19 PNEUMONIA.
NEXT SLIDE PLEASE.
SO PROFESSOR LIDDELOWIOUS
STUDIES HAVE SHOWN THAT SEVERAL
PATHWAYS ARE EMBRACING THE
TRADITIONAL CELL STATUS EMPLOY A
STUDY FROM DR. SHIEWLER'S GROUP
FOUND THAT MANY PATHWAYS RELATED
TO MICROMETABOLISM SUMP
ASOXIDATIVE PHOSPHORYLATION,
EIF2 TARGETS TRANSLATION, TARGET
AND UNFOLDED PROTEIN RESPONSES
ARE UPREGULATED IN TRANSITIONAL
CELL STATE AND IN THE AT1
DIFFERENTIATION.
NEXT SLIDE, PLEASE.
IN ADDITION OUR HUMAN CORE STUDY
SUGGESTS THAT MITOCHONDRIA
COMPLEX 1 MAY BE IN THE
DEVELOPMENT OF PULMONARY
FIBROSIS.
DR. [INDISCERNIBLE] REPORTED
LAST YEAR THAT THE 1 ASSEMBLY
PATHWAY IS ENRICHED IN PATIENT
WITH FAMILIAL PULL MONITOR AREY
FIBROSIS.
NEXT, PLEASE.
SO WE ASKED THE QUESTION DOES
MITOCHONDRIAL METABOLISM CONTROL
LUNG EPITHELIAL STEM PROGENITOR
CELL FATE?
NEXT.
SO WE STARTED WITH A QUESTION,
THAT IS MITOCHONDRIA COMPLEX 1
OR 2, SENTIAL FOR LUNG
DEVELOPMENT.
WE USE SFTPC SYSTEM THAT CRE
COMBINES EMBRYONIC WITH DAY 5
AND WITH [INDISCERNIBLE] AND
SDHD FLOCKS MICE TO COMPLETE
COMPLEX 1 FUNCTION AND COMPLEX 2
FUNCTION RESPECTIVELY EMPLOY SO
THESE MICE, THIS KNOCKOUT MICE
WILL NOT HAVE ANY COMPLEX 1
FUNCTION OR COMPLEX 2 FUNCTION
IN THE DIGITAL LUNG EPITHELIUM
AFTER EMBRYONIC DAY 10.5.
NEXT SLIDE.
SO THE LOSS OF MITOCHONDRIA
COMPLEX 1 FUNCTION BUT NOT
COMPLEX 2 IN THE LUNG EPITHELIUM
RESULT IN POSTNATAL LETHALITY.
SO WHEN WE LOOK AT THIS LUNG
HISTOLOGY ABOUT THIS
[INDISCERNIBLE] KNOCKOUT MICE
WHICH DOESN'T HAVE ANY COMPLEX
LUNG FUNCTION IN THE LUNG
EPITHELIUM, THEY DIDN'T HAVE ANY
INCREASED CELL DEATH OR IMMUNE
CELL FILTRATION BUT RATHER THE
LUNG EPITHELIUM SHOWS VERY
HYPOTROAPIC CELLS.
ESSENTIALLY THEY'RE LONG
EPITHELIAL CELL [INDISCERNIBLE]
WOULD CHANGE.
SO WE WANTED TO KNOW, WHICH PART
OF THE COMPLEX LUNG FUNCTION IS
RESPONSIBLE FOR THIS PHENOTYPE?
NEXT SLIDE.
SO AS YOU SEE IN THE CARTOON AT
THE BOTTOM, COMPLEX LUNG DOES
HAVE ROUGHLY 3 FUNCTION.
IT DOES PUMP UP [INDISCERNIBLE]
THAT EVENTUALLY CONTRIBUTE TO
THE ATP SYNTHESIS, AND
REGENERATE THE REACTIVE OXIDANT
SPECIES AND REGENERATE THE NAD,
SO WE USE THIS NDI 1 PROTEIN,
THIS IS EAST PROTEIN, ESSENTIA
WILY THE EAST VERSION OF COMPLEX
1.
IT DOES THE COMPLEX 1 FUNCTION,
COMPLEX 1, MAMMALIAN COMPLEX 1
FUNCTION WITH ADG REGENERATION
BUT IT DOES NOT PUMP UP THE
PROTON PUMP THEREFORE IT DOESN'T
REALLY ACTUALLY CONTRIBUTE TO
THE ATP SYNTHESIS OR IT DOESN'T
GENERATE THE REACTIVE OXIDANT
SPECIES, SO WE EXPRESS IN THESE
KNOCKOUT MICE TO SEE IF WHICH
FUNCTION OF THE MAMMALIAN
COMPLEX 1 IS CAUSING THIS
PHENOTYPE, IF WE CAN RESCUE THE
LETHALITY OF THIS NDUFS 2
KNOCKOUT MICE BY EXPRESSING NDUF
1, THAT MEANS THAT THE LACK OF
REGENERATION CAUSED THE
PHENOTYPE.
NEXT SLIDE.
SO THE EAST PROTEIN NDI 1
PREVENTS TOTALITY OF NDUFs 2
MICE IN THE PATHOLOGY INDICATING
THAT THE LACK OF OF NAD
REGENERATION, NOT THE SYNTHESIS
OR CHANGES IN REACTIVE OXYGEN
SPECIES CAUSE THE PHENOTYPE OF
THIS COMPLEX 1 KNOCKOUT MICE.
NEXT, PLEASE.
IN ORDER TO UNDERSTAND MORE OF
THE MOLECULAR PHENOTYPE OF THE
ABNORMAL EPITHELIAL CELLS IN
KNOCKOUT MICE, WE PERFORMED AN
RNA SEQ AT POSTNATAL WAY 21 AND
OBSERVED THE TRANSITIONAL CELLS
ARE EXPANDED IN THIS NDUFS2
KNOCKOUT MICE.
THESE CELLS HAVE A SIMILAR
TRANSCRIPTION PROFILE TO THOSE
RECORDED IN MOUSE LUNG INJURY
MODELS EXPRESSING BOTH AT2 ASK
AT1 MARK FIRST AS WELL AS
KERATIN 8 AND CDK N1 A.
NEXT SLIDES.
SO WE GOT THE LIST OF THE
INTEGRATED STRESS RESPONSE, IES
OUR TARGET GENES FROM
DR. [INDISCERNIBLE] AT
[INDISCERNIBLE].
AND CALCULATED THE ISR
ENRICHMENT SCORE WHERE THE
HIGHER SCORE INDICATES A MORE
ENRICHED ISR GEN SIGNAL TOUR,
TRANSITIONAL CELLS IN THIS
KNOCKOUT MICE HAVE VERY HIGH ISR
ENRICHMENT SCORE.
NEXT SLIDE.
SO INTEGRATED STRESS RESPONSE IS
USUALLY AN ADAPTIVE PROCESS THAT
IS ACTIVATED BY STRESS, IT HELPS
THE CELLS ENDURE STRESSFUL
CONDITIONS AND THEN SOMETIMES IT
INDUCE APOPTOSIS.
NEXT.
HOWEVER THE ISR CAN BE
PATHOGENIC IF IT IS HYPER
ACTIVATED OR PERSISTENT.
SO THE SMALL MOLECULE ISR
INHIBITOR ISRI, CAN BRING IT
DOWN TO OPTIMAL LEVELS SO THE
CELLS CAN BECOME A MORE --
THEY'RE THE BEST CONDITION TO GO
THROUGH THE STRESSFUL SITUATION.
YOU WILL HEAR MORE ABOUT THE ISR
INHIBITORS AND THE NEURODISEASE
FROM DR. HELEN MIRANDA TOMORROW.
SO STAY TUNED.
NEXT PLEASE.
SO, WE WANTED TO KNOW IF THOSE
HIGHER ISR THAT WE OBSERVED IN
TRANSITIONAL CELL IS ACTUALLY
THE CAUSAL PATHWAY TO INDUCE
THIS PHENOTYPE SO WE
ADMINISTERED THE ISR INHIBITOR
TO THIS NDUFS2 MICE, AND ABLE TO
MODERATE LETHALITY BY 70%.
THE OUR PATH INDICATE THAT THE
LACK OF NAD REGENERATION CAUSES
A POSTNATAL LETHALITY.
NEXT PLEASE.
SO WE PROPOSED THIS CONCEPTUAL
MODEL OF ATP 2 DIFFERENTIATION,
WE THINK THAT WHEN AT2 CELLS
BECOME TRANSITIONAL CELLS, THE
ADAPTIVE ISR, THE ADAPTIVE ISR
IS ACTIVATED BUT IT IS THE
ACTIVATED WHEN TRANSITIONAL
CELLS DIFFERENTIATE INTO AT1
CELLS.
NEXT.
HOWEVER, WITH THE MITOCHONDRIAL
DYSFUNCTION, SPECIFICALLY
ELEVATED NAD RATIO, THE
PATHOLOGIC ISR IS ACTIVATED AND
THE REMAINS HIGH AND PREVENTING
TRANSITIONAL CELLS FROM BECOMING
AT1 CELLS, WE HAVE A PREVIOUSLY
SHOWN THAT A SMALL MOLECULE
INHIBITOR OF ISR PROMOTES 82-81
DIFFERENTIATION AND IMPROVES
FIBROSIS IN BOTH MICE AND
ASBESTOS-INDUCED FIBROSIS
MODELS.
WE CURRENTLY DON'T KNOW THE
EXACT UPSTREAM AND THE DOWN
STREAM PATHWAYS OF THE PATHOLOGY
OF ISR THAT EFFECTS THE CELL
FATE AND THAT WE ARE USING
GENERATEDITTIC MOUSE MODELS TO
INVESTIGATE THEM.
NEXT SLIDE.
SO THE FUTURE DIRECTION WILL BE
TRYING TO ANSWER THE QUESTION
HOW DID THE INCREASED RESPONSE
CONTROL CELLULAR
DIFFERENTIATION.
AS YOU SEEN AGAIN SEEING AGAIN
IN THIS CARTOON THAT THE ISR IS
ACTIVATED BY 1 OF THE 4 KINASES
UPON STRESS RESULTING IN
PHOSPHORYLATION OF EIF-ALPHA AN
ACTIVATION OF THE ISR,
EVENTUALLY LEADS TO ESSENTIALLY
2 PATHWAY, 1 IS THE REDUCTION IF
GLOBAL PROTEIN TRANSLATION BUT
SIMULTANEOUSLY INCREASE
TRANSLATION OF A CERTAIN
TRANSCRIPTION FACTORS INCLUDING
ATP 4 AND THEN INCREASED THE
TRANSCRIPTION OF HOST OF STRESS
RESPONSE [INDISCERNIBLE] BUT WE
DON'T KNOW -- HOW, WHAT IS THE
FIRST QUESTION IS THAT HOW --
WHAT IS THE UPSTREAM SIGNALS,
HOW DOES THE INCREASE IN NAD
TRIGGER THE ISR.
THANKS TO THE NHLBI THE PROJECT
TO ANSWER THIS QUESTION HAS BEEN
FUNDED SO WE'RE TRYING TO ANSWER
THIS QUESTION WITH A GENETIC
MODELS BUT THE NEXT MORE
IMPORTANT, MORE INTRIGUING
QUESTION IS IS THE ISR NECESSAY
FOR CELL DEFERENTIATION, IN
OTHER WORDS IS IT NECESSARY FOR
CELL DIFFERENTIATION OR
ACTIVATION OF OF THE PATHOLOGIC
ISR IS SUFFICIENT TO CHANGE THE
CELL FATE.
WE HAVE LIKE WE USE SOME GENETIC
MOUSE MODEL TO SHOW THAT
INDICATE THAT THE PATHOLOGIC ISR
ACTIVATION MAY BE DIVISION TO
CHANGE THE CELLS SO HOPEFULLY WE
CAN REPORT THE RESULT AT SOME
POINT IN THE PUBLIC FORM SOON
AND THE NEXT QUESTION IS THIS
CHANGE IN ISR IN CELL FATE DOES
THAT LEAD TO THE LUNG PATHOLOGY?
AND IF SO, WHICH DISEASES DOES
IT CAUSE A FIEB ROSEIS OR AND
WHAT'S THE PATHWAY AND WHICH ARM
OF THE DOWN STREAM PATHWAY OF
THIS ISR IS RELATED TO OR CAUSE
THIS PATHOLOGY, AND ALSO THIS --
BUT AS I MENTIONED IN THE
EARLIER SEVERAL PATHWAYS HAVE
BEEN SUGGESTED TO CONTROL THE
ATP 2 STEM CELL BEHAVIOR BUT HOW
IS IT RELATED TO THIS ISR TO
THOSE OTHER SIGNALING PATHWAYS
AND HOW DOES IT CROSS TALK TO
EACH OTHER AND THEN DO THEY ALSO
CROSS TALK WITH OTHER CELL
TYPES, 2, AND THEN EVENTUALLY AS
A PHYSICIAN SCIENTISTS, WE WANT
TO FIND THE TARGET TO IMPROVE
OUR HEALTH SO IS THIS 1 OF THE
CAN WE IDENTIFY THE TARGET IN
THIS PATHWAY TO IMPROVE THE STEM
AND THE PROGENITOR RESILIENCE SO
THAT WE CAN EVENTUALLY OVERALL
IMPROVE LUNG HEALTH AND THE
RESILIENCE.
SO THAT'S MY OVERALL TALK SO FAR
AND THE NEXT SLIDES IS TO
ACKNOWLEDGE THE PEOPLE WHO WORK
WITH ME AND HELP ME TO DO THE
WORK, SO, I THINK I WILL STOP IN
HERE AND THEN, WE CAN PROBABLY
TAKE THE QUESTIONS IN THE
COMMENTS AT THE END OF THE
SESSION, I BELIEVE.
>> ALL QUESTIONS WILL BE
DISCUSSED IN THE PANEL
DISCUSSION AFTER THE WHOLE
SESSION EMPLOY SO THEN I'M GOING
TO INTRODUCE OUR NEXT SPEAKER
DR. MARIA BASIL, GOOD FRIEND OF
MINE AND ASSISTANT PROFESSOR OF
MEDICINE AT THE UNIVERSITY OF
PENNSYLVANIA.
SHE'S A PHYSICIAN-SCIENTIST WHO
WORKS ON THE COPD I HOPE THAT
SHE'S HERE, YEAH, I'M HERE AND
SO SHE HAS BEEN WORKING ON THE
AIRWAY STEM CELLS ESPECIALLY IN
HUMAN LUNG DEC AND SHE HAS FOUND
REMARKABLE STUDIES TO IDENTIFY
THE NEW TYPES OF AIRWAY
PROGENITOR CELLS AND THE LINK
WITH THE HUMAN PATHOLOGY SO
WE'RE VERY EXCITED TO HEAR ABOUT
HER TALK TODAY AND THEN IT'S ALL
YOURS NOW.
ALL RIGHT, SO THANK YOU FOR THE
INVITATION TO THE SPEAKERS TODAY
AND ORGANIZING THE WORKSHOP,
IT'S A FUN AGENDA AND I'M
LOOKING FORWARD TO HEARING
EVERYONE ELSE SPEAK AND YOU CAN
GO TO THE NEXT SLIDE EMPLOY SO
BECAUSE I WILL JUMP RIGHT IN AND
TELL YOU ABOUT MY FAVORITE
CHRONIC LUNG DEC THEY THINK
COULD BENEFIT FROM RESILIENCE
AND I WANT TO FRAME THIS,
EVERYONE ON THIS TALK KNOWS WHAT
COPD IS, AND EVERYONE ON THIS
TALK KNOWS IT'S A BIG PROBLEM
AND I WANT TO KIND OF FRAME IT
FOR YOU, THE WAY THAT I THINK
ABOUT IT, WHICH IS THAT THIS IS
A REALLY MASSIVE PROBLEM THAT'S
GOING TO BE INCREASING OVER THE
NEXT 20 YEARS.
THIS IS DATA FROM THE CDC AND
THE W. H. O. SHOWING THAT COPD
IS INCREDIBLE LOAMACYY PREVALENT
IN PARTS OF OUR NATION, NOT AN
ELECTION MAP, JUST A MAP OF COPD
PREVALENCE.
PART OF OUR NATION, UP TO 10% OF
THE ALL ADULLS HAVE BEEN TOLD
THEY HAVE SOME FORM OF CHRONIC
PULMONARY DISEASE, AND IT'S THE
LEADING CAUSE OF DEATH WORLD
WIDE AND IT'S NOT JUST A
SMOKER'S DISEASE, WE'VE HEARD
THAT THAT THERE'S LOTS OF OTHER
IDEOLOGIES FOR LUNG DEC, LIKE
AIR POLLUTION, BIOMATTER AND SO
THIS NUMBER WILL GROW EMPLOY
NEXT SLIDE ISSUE PLEASE.
SO I WANT TO JUST WANT TO FRAME
IT FOR THIS AUDIENCE ABOUT WHY
WE DO THE WORK THAT WE DO THAT
THIS IS A BASIC SCIENTIST, I
THINK ABOUT CELLS AND
TRANSCRIPTION FACTORS AND CELL
DIVISION A LOT BUT THIS IS
REALLY A SOCIAL JUSTICE ISSUE
AND I THINK IT'S WORTH BRINGING
UP AS WE DO THE WORK THAT WE DO,
THAT THIS IS A DISEASE THAT HAD
DISPROPORTIONATELY AFFECTED
PATIENTS OF COLOR ACROSS THE
UNITED STATES AND HAS BEEN
UNDERDIAGNOSED IN THESE PATIENTS
AND AS MUCH AS I LIKE TO SEE
FEMALES CATCH UP TO MALES AND
ALL SORTS OF WALK ANDS METRICS
OF LIFE, THIS IS NOT REALLY THE
GRAPH I WANT TO DISPLAY WHICH IS
OVER THE LAST 40 YEARS, WOMEN
HAVE SUCCESSFULLY CAUGHT UP TO
MEN IN PREVALENCE OF MORTALITY
FROM COPD.
AND SO THIS IS LIKE I SAID AN
ACCELERATING PROBLEM.
SO WHY DO I THINK ABOUT COPD.
WELL IT'S REALLY A DEGENERATIVE
DISEASE.
YOU DON'T NEED TO BE A
PATHOLOGIST TO BE ABLE TO LOOK
AT THAT LUNG AND SAY IT DOESN'T
LOOK GOOD.
SO UPPER LOW PREDOMINANT TISSUE,
DESTRUCTION LEADING TO THESE
HOLES IN THE LUNG.
NEXT SLIDE.
AND THE WAY I THINK ABOUT COPD
AS A STEM CELL OR PROGENITOR
CELL BIOLOGIST AS SOMEONE WITH A
BACKGROUND THIS DEVELOPMENTAL
BIOLOGY IS HOW IT EXISTS IN
CONTRAST TO THE FORMS OF INJURY
FOR EXAMPLE WE JUST HEARD ABOUT
FROM DR. HAN, SO THIS IS A CAT
SCAN OF A PATIENT WITH COPD ON
THE TOP AND VIRAL PNEUMONIA ON
THE BOTTOM EMPLOY SO BOTH OF
THESE DISEASES ARE MARKED BY
PROFOUND DISRUPTION OF TISSUE,
DISRUPTION OF THE GAS EXCHANGE
FUNCTION OF THE LUNG AND IN BOTH
OF THESE THIS ALVEOLAR
DISRUPTION AND COMPLETELY
DESTROYED.
BUT WHAT'S STRIKING TO ME AS A
STEM CELL DEVELOPMENTAL
BIOLOGIST IS THAT PATIENTS WITH
VIRAL PNEUMONIA, THEY END UP
INTUBATED IN OUR INTENSIVE CARE
UNIT AND 6 WEEKS LATER THEY WERE
CRITICALLY ILL IN THE UNIT, A
MONTH OR SO LATER THEY WALK IN
THE CLINIC AND THEY GET A CAT
SCAN ON THE BOTTOM, THEIR LUNG
HAS BEEN A WAY TO BE RESILIENT
TO GET THROUGH THIS INJURY AND
HEAL ITSELF ON THE OTHER SIDE
AND NOW MAYBE THEY'RE NOT
WEARING OXYGEN AND THEY'RE
RECOVERING FROM THIS RETINAL
LOCATIONRIMENTAL INJURY.
COPD DOES NOT WORK THAT WAY.
NO 1 HAS SEEN A PATIENT WITH
COPD WHO'S ALVEOLAR TISSUE HAS
GOTTEN BETTER.
NEXT SLIDE PLEASE.
AND SO WHEN I REALLY WANTED TO
THINK ABOUT HOW I WOULD APPROACH
THIS PROBLEM, I BECAME VERY
INTERESTED IN LIGHT OF THAT IN
THE DISTAL HUMAN AIR WAYS AND
THAT'S BECAUSE THERE'S BEEN A
PLETHORA OF DATA CULMINATING
REALLY IN THIS 2018 GROUP FROM
BRITISH COLUMBIA AND THE DATA
WENT ALL THE WAY BACK TO THE
PRESSURE VALIUM STUDIES IN THE
1960S, SUGGESTING THAT COPD WAS
A DISEASE THAT STARTED FIRST IN
THE DISTAL RESPIRATORY AIR WAYS
OF THE HUMAN LUNG BUT THIS
INJURY MAY PROCEED THAT SORT OF
SIN QUANON OF STIGMA, THAT THING
WE THINK OF OF EMPHYSEMA AND THE
DISTAL LUNGS.
BUT MICE LACK THIS REGION OF THE
LUNG, THEY DON'T HAVE THIS
TERTIARY STRUCTURE AND I'M SURE
MOST PEOPLE ON THIS CALL KIND OF
KNOW THAT DISTAL AIRWAY
STRUCTURE.
NEXT SLIDE.
SO IN THINKING ABOUT HOW I WAS
GOING TO START THINKING ABOUT
THIS, WE AND OTHERS AS WELL,
INCLUDING GOOD FRIEND OF MY
[INDISCERNIBLE] MURPHY WHO WAS
ON THE CALL, WITH
[INDISCERNIBLE] HO I KNOW IS IN
THE CALL BECAUSE HE'S IN THIS
SECTION, SO BEFORE WE KNEW EACH
OTHER WE WERE WORKING ON A
SIMILAR QUESTION HAND IN HAND
AND WE CAME TO VERY SIMILAR
CONCLUSIONS WHICH IS THAT
THERE'S A POPULATION OF
SECRETATTORY CELLS THAT LINE THE
HUMAN LUNG, THEY CALL THEM
TRBSCS, WHICH YOU CAN SEE I
MISSPELLED AT THE BOTTOM OF THE
SLIDE HERE WE CALL THIS
RESPIRATORY SECRET CELLS AND
THESE ARE SITTING IN THIS
RESPIRATORY AIR WAY OR THIS PART
OF THE LUNG THAT A COMPLEX
TERTIARY STRUCTURE DISTINCT TO
MAMMALS AND WE IDENTIFIED THIS
RARE POPULATION OF CELLS THAT
ARE AT2 LIKE BUT EXPRESS AIRWAY
MARKERS WHICH CALLED AT0S AND WE
REFER TO AS DOUBLE POSITIVE AT2
CELLS.
NEXT SLIDE.
AND I THINK IMPORTANTLY AS WE
THINK ABOUT HOW WE'RE GOING TO
APPROACH THESE QUESTIONS GOING
FORWARD, HOW WE WANT TO MODEL
THIS, BOTH OF US TOOK DIFFERENT
APPROACHES TO LOOK AT THIS
ACROSS SPECIES, SO I'M SHOWING
YOU HERE SINGLE CELLS
[INDISCERNIBLE] WORK ON MACAQUES
AND WE WORKED TO IDENTIFY THE
RESPIRATORY BRONCHIAL STRUCTURES
IN FERRETS AND IDENTIFYING AND
SHOWING THEY HAVE THE SAME CELL
POPULATIONS.
NEXT SLIDE, PLEASE.
SUBSEQUENT TO THIS, WHAT WE
REALLY WANTED TO DO WAS LOOK AT
THESE CELLS, AND ASK WHAT WAS
THEIR FUNCTION IN THE DISTAL
HUMAN LUNG.
AND FOR THIS, I TURN TO
COLLABORATE WITH DARYL COTTON
AND ANDREW HOPKINS AND ANDREW
WILSON AND THEIR WHOLE TEAM UP
THERE TO ASK THE QUESTION, TO
TEST THE HYPOTHESIS RATHER THAT
RAS CELLS COULD ACT AS A CELL
PROGENITOR.
THERE ARE SEVERAL REASONS WE
THOUGHT THIS MIGHT BE TRUE AND
SO WHAT I'M SHOWING IS A
PLETHORA OF INFORMATION
CONDENSED INTO 1 IMAGE WHICH IS
THAT WE WERE ABLE TO GROW THESE
RAS-LIKE CELLS DERIVE FRIDAY ES
CELLS AND AN ESL CULTURE SYSTEM
AND AIRWAY MEDIA AND THEN IF WE
ASKED COULD THESE CELLS
TRANSITION TO ALVEOLAR LIKE
CELLS USING ALVEOLAR CONDITIONS
THAT HAD BEEN DEFINED BY
DERILLEGALS AND ANG LEAN JACOBS
WE WERE ABLE TO SHOW THESE CELLS
WITHIN 48-72 HOURS COULD RAPIDLY
TURN UP THEIR SECRETATTORY CELL
FATE AND TURN ON THIS ALVEOLAR
LIKE PROGRAM AND THEY WERE ABLE
TO EXPRESS THE FACT IN PROTEIN C
AND ALSO PROCESS SURFACTIN
PROTEIN C THAT MADE IT LOOK LIKE
THEY WERE ACTING LIKE AT2 CELLS.
SO OF COURSE WE WANT TO LOOK AT
THIS IN LUNG DEC AND SO WHAT ARE
THE DISEASES THAT WOULD BE THE
MOST INTERESTED IN FOCUSING ON?
WELL OF COURSE I TOLD YOU
EARLIER THAT I'M REALLY
INTERESTED IN COPD OR CHRONIC
OBLIGATIONS JUCTIVE PULMONARY
DISEASE EMPLOY THERE ARE A LOT
OF OTHER DISEASES THAT OCCUR IN
THIS REGION OF LUNG THAT WORTH
CONSIDERING, BRONCHITEIS OBLIT
RANS SYNDROME, WHICH I WILL COME
BACK TO, AND LUNG TRANSPLANT, IF
YOU CAN MAKE IT TO A YEAR WITH
THE LUNG TRANSPLANT, THE MOST
LIKELY CAUSE OF BOS, AND THERE
ARE FORMS OF INTERSTITIAL LUNG
DISEASE, SOME DATA SUGGIEST
MAYBE EVEN IPF ORIGINATES IN
THIS REGION AND WORK FROM JIM
[INDISCERNIBLE]'S GROUP
SUGGESTED IT'S A DISEASE THAT WE
WILL ALL COME TO ENDURE SOME OF
US EARLIER THAN OTHERS IN AGING
AND IT ALSO AFFECTS THIS REGION
OF THE LUNG.
NEXT SLIDE, PLEASE.
SO WE STARTED BY LOOKING AT
CHRONIC OBLIGATIONS JUCT UF
PULMONARY DEC SO I WILL START
WITH END STAGE DISEASE, SO WEEZ
ARE GOING ON FOR TRANSPLANTATION
FURTHER FOR COPD, AND PATIENTS
WITH COPD, AND THE SEVERITY TO
WHICH THEY REQUIRE LUNG
TRANSPLANT, YOU CAN SEE WE SAW
AN INCREASE IN THE 3A 2 POSITIVE
CELLS AND ALSO I CALLED AT0
CELLS, AND CAN SEE THESE CELLS
ARE IDENTIFIED BY SUPPRESSION OF
SURFACTIN PROTEIN C, THEY LOOK A
LOT LIE AIRCRAFT T2 CELLS BUT
THEY KRE TINS CRETE
IMMUNOGLOBULIN 2, WHICH MAKES US
THINK THEY HAVIZE CHEESE
CHARACTERISTICS.
NEXT SLIDE.
SO IT'S IDENTIFYING SINGLE CELL
SEQUENCING CAN WE IDENTIFY THEM
IN SECTIONS FROM DIFFERENT
PATIENTS, SO AGAIN, IN PATIENTS
WITH END STAGE COPD WHO ARE AT
THE POINT OF TRANSPLANTATION,
COMPARED TO HEALTHY AGED MATCH
CONTROLS WHO WERE NEVER SMOKERS
WE SEE THESE DOUBLE POSITIVE OR
OT0 LIKE CELLS ARE INCREASED IN
THIS PATIENT POPULATION.
AND WHAT WE REALLY WANT TO GET
AT OF COURSE IS THIS SOMETHING
THAT'S EVOLVING OVER INJURY, IS
THIS PART OF DEVELOPMENT OF
DISEASE, SO NEXT SLIDE, PLEASE.
WE TOOK AN APPROACH WHEREY WE
USED NORMALLY LUNG DONORS AND
ASSAY, USED BASED ON REPORTED
SELF-REPORTED CIGARETTE SMOKE
EXPOSURE IN PACK YEARS SO
THERE'S A LOT OF CONFOUNDERS
HERE, I UNDERSTAND, BUT BASED ON
REPORTED, SELF-REPORTED P A C
REPORTERS IN DIFFERENT REGION OF
THE LUNG, WE ASKED WHAT WAS THE
PREVALENCE OF THESE CELLS ACROSS
THE LUNG.
AND AS WE -- WE OBSERVED WAS
THAT AS PATIENTS HAD AN
INCREASED AMOUNT OF SIGNATURE,
THEY ACCUMULATE MORE OF THESE
DOUBLE POSITIVE CELLS.
NEXT SLIDE, AS I MENTIONED THAT
ASSAY AND THAT EXPERIMENT IS
INCREDIBLE LOAMACY CONFOUNDED BY
ALL THE LIMITATIONS THAT COME
WITH HUMAN SUBJEKSENTIOUS
SPECIALLY DONATED LUNG RESEARCH
AND SO WE TURN THEN TO A
COLLABORATION WITH
[INDISCERNIBLE], AND
[INDISCERNIBLE] DOWN AT THE
UNIVERSITY OF ALABAMA WHERE WE
LOOKED AT THE FERRET MODEL THAT
[INDISCERNIBLE] HA PUBLISHED
MANY YEARS AGO, IF YOU GIVE A
CIGARETTE OR MANY SIGNATURESIETS
THEY GEP A LOT OF PATIENTS THAT
LOOK LOOK THE PATIENTS I TAKE
CARE OF, WEIGHT LOSS, AIRWAY
OBSTRUCT, IMMUNE INFILTRATE, AND
WE FOUND THERE WAS A INCREASE OF
DOUBLE POSITIVE CELLS IN THIS
DISEASE.
SO THIS SUGGIESTED TO US THIS
MODEL SYSTEM MAY BE
RECAPITULATING EPITHELIAL
RESPONSES THAT ARE ANALOGOUS TO
OUR PATIENTS AS WELL.
NEXT SLIDE.
SO I'LL TURN TO ANOTHER CHRONIC
DISEASE, JUST TO POINT OUT SOME
OF THE OTHER STUFF THAT WE'VE
BEEN WORKING ON IN THIS ARENA OF
THE DISTAL LUNG AND LOOKING AT
THE RESPONSE OF THE DISTAL LUNG
IN CHRONIC DISEASE EMPLOY SO
THIS IS DATA FROM
BRONCHIALALATED RELATED
SYNDROME, AND HAS 2 MAIN CAUSES
AND A FEW OTHER MORE RARE CAUSES
INCLUDING POST LUNG TRANSPLANT
AND POST HEMATOPOIETIC
TRANSPLANT.
AND YOU SEE WE COMBINED PATIENTS
FROM BOTH SYNDROMES ALONG WITH A
SERIES OF AGE AND GENDER MATCHED
CONTROLS.
AND WE CAN SEPARATE THESE BY
THEIR IMMUNE COMPONENT AND THEIR
NONIMMUNE COMPONENT ASSAY
SEPARATELY.
NEXT SLIDE.
SO WHAT WE WERE REALLY -- WE
LOOKED AT A BUNCH OF DIFFERENT
THINGS AND 1 OF THE THINGS WE
WERE REALLY STRUCK BY IS THAT IN
THE DISTAL AIRWAY EPITHELIUM AND
IN PARTICULAR IN THE AIR WAY
OPERATING GLOBALLIULATION AND IN
THE ALVEOLAR TYPE 2 POPULATION
WE SEE DRAMATIC INCREASE IN
SEVERAL FACTORS 1 OF WHICH IS
CXCLA OR INTERLEUKIN 8, I WILL
NOT PLAY THE ROLE OF AN
IMMUNOLOGIST TODAY BUT 1 OF THE
THINGS THAT THIS HAS BEEN SHOWN
TO DO IS RECRUIT SUBSETS OF
IMMUNE CELLS INCLUDING
NUTRIFILLS AND T-LYMPHOCYTES AND
THERE'S BEEN A LOT OF WORK ON
THE ROLE OF IMMUNE POPULATIONS
AND OBLIT RANS, AND NEXT SLIDE,
SO WE WANTED TO LOOK AT WHAT
WERE THE CELL AND LYMPHOCYTE
SUBSETS IN PARTICULAR, THAT WERE
RECRUITED INTO THE DISTAL LOG
AND THERE HAD BEEN A LOT OF WORK
BEFORE IN BALL STUDIES FROM MARK
SNYDER AND OTHERS LOOKING AT
SIMILAR QUESTIONS AND WHAT WE
SEE IS THAT IN THE DISTAL AIRWAY
IMMUNE CELLS, IN OUR PATIENTS
WITH END STAGE BRONCHIALITEIS,
SO THESE ARE PATIENTS WHO HAVE
BRONCHIALITEIS WE SEE
UPREGULATION IN MANY DIFFERENT
MARKERS INCLUDING EXHAUSTION
EFFECTOR AND THESE MARKERS IN
BLUE CALLED TRM MARKERS OR
TISSUE RESIDENT MEMORY MARKERS.
NEXT SLIDE.
SO WE USE MULTIPLEX
IMMUNOHISTOCHEMISTRY, I'M
SHOWING 4 IN THE LUNG, WE CAN
SEE THIS IS A SEGMENT OF A
PATIENT FROM BRONCHIAL OBLIT
RANS, WHERE YOU SEE THE
INFILTRATES IN THE PARTS OF THE
LUNG AND YOU CAN SEE THE TRM
MARKERS OF INHIBITORY AND
EFFECTED MARKERS AND THIS CO
EXPRESSION IS SORT OF 1 OF THE
CLASSICAL DEFINING FEATURES OF
THESE TRM SUBSETS.
NEXT SLIDE.
SO 1 THING WE'VE BEEN DOING IS
NOT JUST COLLECTING FROM
PATIENTS WITH BRONCHIALITEIS,
BUT ALL SORTS OF LUNG ISSUES, SO
I'M SHOWING TISSUE RESIDENT
MEMORY SIGNATURE THAT WE DERIVE
FRIDAY OUR PATIENT SASMEMS AND
COMPARED IT ACROSS A LARGE
SMATTERING OF DISEASES ACROSS
OUR COHORT HERE AND ASKING HOW
DOES THIS TISSUE MEMORY COMPARE
IN CD8S AND C, DID 4S ACROSS
MANY DIFFERENT LUNG DISEASES AND
WHAT I HOPE YOU CAN SEE IS THAT
WHILE TRM CELLS HAVE BEEN
IMPLICATE INDEED A LOT OF
DISEASES, THAT DISEASE SIGNATURE
IS VERY HIGH IN OUR PATIENTS
WITH BRONCHIALITEIS OBLIT RANZ
AND WE FIND THIS INTERESTING
THAT'S WORTH FOLLOWING UP ON.
AND SO NEXT SLIDE.
WITH THAT, I WILL JUST TAKE A
SECOND TO SAY THAT COPD AND
OTHER DISEASES OF THE RESPIRATOR
AIRWAYS ARE A MAJOR IMPACT AND
COPD IN PARTICULAR DEFINED BY
FAILURE OF REGENERATION IN THE
RESPIRATORY AIRWAY ANDS THERE
ARE UNIQUE POPULATIONS HERE AND
LARGE ANIMAL MODEL SYSTEMS MAY
BE REALLY BENEFICIAL FOR
ADVANCING OUR KNOWLEDGE IN THIS
AREA OF PULMONARY
PATHOPHYSIOLOGY.
NEXT SLIDE.
THAT'S WHO DID THE WORK
INCLUDING THE PICTURE DOWN THERE
OF PATRICK MELLORS, HE IS A
FELLOW HERE AT PENN, AND THE
FUNDING AND A PICTURE OF GROUP
AT A SURPRISE BIRTHDAY PARTY.
WITH THAT I AM HAPPY TO TAKE
QUESTIONS IN THE DISCUSSION
SESSION.
THANK YOU VERY MUCH.
>> THANKS SO MUCH.
NOW MOVING ON TO OUR NEXT
SPEAKER DR. JAYARAJ RAYAGOP A L,
PROFESSOR HARVARD MEDICAL
SCHOOL.
YAY IS A PRINCIPAL VRGS AT MGH
AND HE'S ALSO A PRINCIPLE
INVESTIGATOR AT THE HARVARD STEM
CELL INSTITUTE.
HE'S ALSO AFFILIATE MEMBER FOR
BROAD IT'S CONSTITUTE AND A
SCHOLAR.
JAY MAY A SIGNIFICANT IN THE
AREA OF STEM CELL AND TODAY HIS
TALK IS NEW AREA CELL TYPES,
HELOC CELL, FOR RESISTANT
RESERVOIR OF UNIQUE PLASTIC STEM
CELL.
THANKS A LOT, JAY.
>> THANKS SO MUCH I ENJOYED THE
TALKS A LOT AND UNDERSTAND WHAT
THE CONFERENCE IS ABOUT, THE
JUXTO POSITION OF THE KEY NEAT
SPEAKERS FRAMED WHAT IT IS.
I WILL TALK TO 1 SMALL ASPECT OF
IT.
IF YOU COULD FLIP TO THE NEXT
SLIDE.
A WHILE BACK, A GROUP AND MINE
REDEFINED THE AIRWAY EPITHELIUM
FROM SOMETHING THAT LOOKED
RELATIVELY SIMPLE TO SOMETHING
THAT WAS A LOT MORE RICH AND
COLORFUL EMPLOY IF YOU GO TO THE
NEXT SLIDE, YOU WILL SEE WE WERE
ABLE TO RESOLVE THE LINEAGE
HIERARCHY OF THE MOUSE TRACHEAL
EPITHELIUM AND THE HUMAN IS
STILL NOT QUITE DONE AS IT'S
MORE DIFFICULT BUT YOU CAN SEE A
LOT OF THE EMPHASIS IN THE
ORIGINAL PAPER WAS ON THE
DESCRIPTION OF THESE NEW AIRWAY
CELL TYPES, THE IONOSIGHT
CAPTURING MOST OF THE ATTENTION.
I THINK [INDISCERNIBLE] WILL
TALK ABOUT THAT LATER BUT THE
CELL THAT MOST INTRIGUED ME WAS
THE KRT13 POSITIVE CELL THAT
SEEMED TO BE INTERMEDIATE
BETWEEN BASAL CELLS AND
SECRETATTORY STEM CELLS.
NEXT SLIDE, PLEASE EMPLOY I WILL
STEP BACK AND TAKE YOU BACK TO
THE TEXTBOOK BECAUSE THE AIRWAY
EPITHELIUM IS AS MANY OF YOU
WILL REMEMBER, IS THE SEEN O
QUANON PSEUDOSTRATIFIED ELTHEL
NUM AND THE 2 LAYERS NUCLEI DID
NOT MEAN THAT EVERY CELL DIDN'T
TOUCH THE CELL BASEMENT
MEMBRANE.
SO IT'S A VERY PARTICULAR KIND
OF EPITHELIUM.
ONE OF THE CLASSIC OF THEM
INCLUDING SQUAMOUS AND
[INDISCERNIBLE].
IF YOU GO TO THE NEXT SLIDE, THE
OTHER INTERESTING THING I WOULD
OBSERVE IS THE AIRWAY HAS VERY
FEW REACTION MECHANISMS AND WILL
2 BIGGEST 1S WHEN YOU LOOK AT
THE TISSUE, ARE THE GOBLET IN
THE MITSD AND SQUAMOUS PRODICTED
ON THE RIGHT.
AND RAVI, IF HE TOLD YOU ABOUT
COPD, HE WOULD SAY THAT BOTH OF
THESE PATHOLOGIES OCCUR IN 1
DISEASE AND THAT'S ACTUALLY TRUE
OF ASTHMA AND MOST INFLAMMATORY
AIRWAYS DISEASES BUT THERE HAS
BEEN ANY RESOLUTION TO WHY THAT
WOULD BE.
SO IF YOU GO TO THE NEXT SLIDE.
THE REASON I LIKED THOSE
CYTOKERATIN 13 CELLS EVEN MORE
THAN [INDISCERNIBLE] AT THE TIME
WAS BECAUSE THEY ACTUALLY
OCCURRED IN STRUCTURES.
AND WE CALL THOSE STRUCTURES
HILLOCKS, ON THE TOP AND ON THE
BOTTOM YOU WILL SEE A
[INDISCERNIBLE] WHERE THERE ARE
THESE CELLS THAT ARE DISTRIBUTED
AMONGST THE GREEN CILIATED CELLS
AND ON THE RIGHT LOWER PANEL YOU
WILL SEE THEY'RE CONNECTED BY
DESMODEL CITIZENSOMES ON EM.
NEXT SLIDE.
AND THESE HILLOCKS ARE FURTHER
MORE DESSERS FOR ALL THOSE RARE
CELL POPULATIONS WE DESCRIBED.
NEXT SLIDE.
SO WHEY WANT TO LEAVE YOU WITH
FOR THE TIME BEING IS THAT THE
PSEUDOSTRATIFIED EPITHELIUM IS
INDEED VERY COLORFUL WITH ALL
THESE NEW RARE CELLS WHOSE
DEFINITION AND FUNCTIONS REALLY
ARE ONLY BEGINNING TO BE
UNDERSTOOD BUT WITHIN THAT, THE
HILLOCK IS A DISCREET EPITHELIUM
STRUCTURE THAT REALLY SEEMS TO
HAVE ITS OWN PROPERTIES.
NEXT SLIDE.
SO WE WERE VERY CONCERNED
BECAUSE THE HILLOCK APPEARED TO
HAVE THAT KIND OF SQUAMOUS
MORPHOLOGY ON THE SURFACE CELLS
THAN I SHOWED YOU IN THAT ONPH
OS IMAGE.
BUT THE AIRWAY EPITHELIUM IS NOT
SUPPOSED TO HAVE THOSE CELLS
UNLESS THERE'S A METAPLASTIC
RESPONSE AND AS I SAID THE
TEXTBOOKS HAVE DEFINED THIS ASA
AGHT WAY, BUT THE HILLOCKS WORK
ON THIS SO IF YOU HIT CLOCK ON
THIS, YOU WILL SEE WE LABELED
INDIVIDUAL HHILLOCKS, BUT IT
LOOKS LIKE THE EPITHELIUM HAS
THE [INDISCERNIBLE] CELLS, SO
THE HILLOCKS ARE STRATIFIED
SQUAMOUS CELLS EXISTING IN THE
NORMAL AIR WAY OF THE MOUSE.
NEXT SLIDE.
AND WHAT I WANT TO POINT OUT IN
THIS CARTOON, IF YOU LOOK AT THE
MARKER OF THE CYTOKERATIN 13,
THERE'S A GRADIENT BUT EVEN THE
BASIL STEM CELLS WITHIN THE
HILLOCK EXPRESS KRT13, SO THOSE
SEEM TO BE DIFFERENT THAN THE
STEM CELLS OF THE STRATIFIED
EPITHELIUM.
NEXT SLIDE.
THE OTHER INTRIGUING PROPERTY
WHICH WE HAVE STILL NOT RESOLVED
IS THAT HILLOCKS DON'T SEEM TO
BEHAVE LIKE OTHER DEVELOPMENTAL
STRUCTURES.
FOR EXAMPLE WHAT I MEAN BY THAT
IS THEY ARE NOT BILATERALLY
SYMMETRIC.
IN DIFFERENT ANIMALS YOU WILL
SEE DIFFERENT PATTERNS OF THESE
HILLOCKS, AND ON THE RIGHT YOU
WILL SEE A CARTOON, WHERE THEY
SEEM TO OCCUR IN 3
CHARACTERISTIC REGIONS BUT THEY
ARE NOT STEREOTYPED.
IT'S STILL A PUSLE, WE DON'T
HAVE A RESOLUTION TO, BUT MAY
CARRY IMPLICATIONS AND DISEASE
AND WE CAN SPECULATE ABOUT THAT
LATER.
NEXT SLIDE.
BUT WHAT'S THE FUNCTION?
BECAUSE THEY WERE SQUAMOUS AND
STRATIFIED THEY REMIND US OF
SKIN AND THEY HAVE PLENTY OF
[INDISCERNIBLE], SO WE WONDERED
IF THEY HAVE A BARRIER FUNCTION.
SO HERE'S A ANIMAL TREATED WITH
NAPH ALENE, AND YOU WILL SEE
THERE'S VERY FEW LEFT AFTER
NAPTHALENE, BUT THERE'S SOME UP
THERE IN THE GLANCE WHERE THERE
IS SUPPOSED TO BE A STEM CELL
RESERVE POPULATION BUT
NONETHELESS, YOU SEE THE
HILLOCKS ARE ALMOST THE ONLY
REMAINING STRUCTURES ON THIS AIR
WAY EPITHELIUM.
NEXT SLIDE.
EVEN MORE REMARKABLY, THEY
SURVIVE IN A FREEZER OVER
MULTIPLE TIMES ACTUALLY AND
ABSENCE OF ANY KIND OF DMSO, SO
THESE STRUCTURES ALSO PHYSICALLY
RESIST AMAZING FORMS OF INJURY.
NEXT SLIDE.
AND HERE THEY RESIST FLU
INFECTION, THEY WON'T BE SEEN BY
THE MTB IN FLU, SO THEY ARE
RESILIENT.
NEXT SLIDE.
IN FACT, THEY ARE RESILIENT TO
EVERYTHING WE CAN THROW AT THEM,
I'VE ALREADY SHOWN YOU FLU,
FREEZE THAT YOU, AND NAPH
THALENE, BUT WITH WE THROUGH
MORE AT IT, THE STRUCTURES
RESISTED TO SOME DEGREE.
I WOULD POINT OUT THEY ALSO
RESIST PROTEASE DIGESTION, THIS
IS IMPORTANT NOT BECAUSE IT'S
SOMETHING THAT YOU OFTEN
ENCOUNTER, ALTHOUGH MANY
ALLERGIENS DO HAVE PROTEASES BUT
I PUT IT HERE BECAUSE MANY OF
OUR SINGLE CELL DATA SETS MAY
NOT CONTAIN HILLOCKS BECAUSE
AFTER SINGLE CELL DISASSOCIATION
THEY'RE LIKELY TO BE
UNDERREPRESENTED.
NEXT SLIDE.
SO WHAT DO THEY DO?
HERE IS WHAT THEY DO.
IF YOU TAKE 1 OF THOSE HILLOCKS
AND IF YOU TAKE 1 OF THOSE,
DAMAGE THEM AND PUT THE TRACHEAL
EXPLANT INTO TISSUE CULTURE, CAN
YOU SEE WHAT THEY DO.
THEY RESURFACE THE AIRWAY
EPITHELIUM.
NEXT SLIDE.
WE PERFORMED A LINEAGE TRACE,
WHERE YOU SEE THE TRACE
INDICATED BY TOMATO AND YOU
WEIGH THE COURSE OVER
REGENERATION, YOU SEE THAT THE
TD TOMATO OVERTAKES THE AIRWAY
EPITHELIUM.
NEXT SLIDE.
AND INDEED, NOT ONLY DOES THE
HILLOCK REMAKE THE ENTIRE
PSEUDOSTRATIFIED EPITHELIUM SO
THE NONLOCK MAKES THE
EPITHELIUM, IT EVEN MAKES THIS
PANOPLY OF THESE CELLS.
NEXT SLIDE.
WHAT'S MORE REMARKABLE IS IF YOU
DO A CLONAL LINEAGE TRACING OR
RAINBOW LINEAGE TRACING, I WOULD
SAY, THE HILLOCKS THROW OFF
THESE VERY, VERY LARGE CLONES OF
CELLS THAT BECOME
PSEUDOSTRATIFIED AIRWAY
EPITHELIUM AND I SHOWED YOU THAT
BOX BECAUSE IT'S APPROXIMATELY
THE SIZE OF THE [INDISCERNIBLE].
SO THERE ARE CELLS THAT CAN
UNDERGO MASSIVE EXPANSION EMPLOY
NEXT SLIDE.
SO, THE OTHER THING I POINT OUT
IS HILLOCKS LOOK A LOT LIKE
SQUAMOUS METAPLASIA THAT'S BEEN
NOTICED FOR A LONG TIME.
THE BEST DEFINED MOLECULARLY
FORM OF SCWAIPOUS METAPLACIA IS
VITAMINA DEFICIENCY.
NEXT SLIDE.
I WILL SKIP THE FINDING THAT I
DIDN'T -- I DIDN'T HAVE TIME TO
SHOW YOU THE ACTUAL DATA BUT IN
THE SETTING OF VITAMIN A
DEFICIENCY IN A MOUSE, HILLOCKS
GET BIGGER.
SO THE SOURCE OF VITAMIN A
DEFICIENCY ASSOCIATED SQUAMOUS
METAPLACIA IS HILLOCKS.
NOW I CAN'T MAKE THAT STATEMENT
ABOUT OTHER INJURIES, I CAN'T
MAKE THAT STATEMENT ABOUT
SMOKES, ASTHMA OR COPD BUT IT
REMAINS TO BE DETERMINED WHAT
THE RULE OF THE HILLOCK IS IN
THOSE DISEASES, BUT WHAT IS
INTERESTING IS WHEN YOU COMPARE
HILLOCK BASAL CELLS TO NORMAL
BASIL CELLS, HILLOCK BASIL CELLS
SHOW DEMONSTRATION BUT THEY
METRICS TAB O LIES RETINOIC ASIT
DIFFERENTLY.
NEXT SLIDE.
AND IF YOU SUBSIDIARY CONSTITUTE
BASAL STEM CELL, BUT THE CELLS
ON THE LEFT RESIST THIS.
NEXT SLIDE.
AND HERE HUMAN HILLOCKS EXIST.
WHAT'S INTERESTING INTEREST
THESE HUMAN HILLOCKS IS ACTUALLY
THEY WOULD BE ENTIRELY MISSED BY
PATHOLOGISTS, THEY WOULD NOT
RECOGNIZE THIS STRUCTURE AS
PATHOLOGIC BECAUSE IT DOESN'T
QUITE QUALIFY FOR THE DEFINITION
OF SQUAMOUS METAPLACIA.
SO PATHOLOGISTS SEE THESE THINGS
BUT THEY DON'T KNOW WHAT TO MAKE
OF THEM.
NOW ALONGSIDE SARRA LYNN'S LUNG
MAP INITIATIVE, WE WILL BE ABLE
TO MARK THESE HILLOCKS AND USING
TRANSCRIPT ORDER OF MICRONSICS
DEFINE WHAT THEY ARE.
NEXT SLIDE.
SO WHY CONTINUE TO STUDY THE
HILLOCK, I THINK THERE'S A
COUPLE OF REASONS, THE REASON I
DON'T HAVE A SLIDE FOR IS
EXACTLY WHAT YOU HAVE DEDICATED
THIS CONFERENCE TOWARDS WHICH IS
RESILIENCE.
THE HILLOCKS SEEM TO RESIST
EVERY INJURY WE CAN THROW AT
THEM AND THEN THEY EXPAND.
AND INDEED, RECENTLY WE WERE
TALKING TO SAM JAMES FROM UCL
AND PETER CAMPBELL FROM THE
SANGER AND THEY'VE IDENTIFY
THESE CELLS THAT EXPANDED THAT
CAN BE SEEN AFTER THE CESSATION
OF SMOKING.
SO THE QUESTION IS, ARE THOSE
HILLOCK CELLS OR ARE THEY
SOMETHING ELSE.
BUT THE OTHER REASON TO STUDY
HILLOCKS IS THAT, YOU KNOW THERE
IS THIS CLASSIC METAPLASTIC
SEQUENCE, SO MAYBE THE HILLOCKS
HELP IN TERMS OF RESILIENCE, BUT
AT THE SAME TIME, MAYBE HILLOCKS
ARE ACTUALLY THE ORIGIN FOR THE
SQUAMOUS METAPLACIA, DYSPLASIA
AND CARCINOMA INSITU THAT 1
FINDS IN SQUAMOUS CELL CANCER OR
MAYBE NOT.
THEY COULD EITHER BE A SOURCE OF
RESILIENCE OR A SOURCE OF CANCER
OR THEY COULD INDEED BE BOTH, WE
SIMPLY JUST DON'T KNOW YET.
NEXT SLIDE.
SO I'LL END EARLY, I THINK I
SAID WHAT I HAD TO SAY BRIEFLY,
SO THE QUESTIONS REALLY NOW ARE
WHAT IS THE IMPORTANCE OF THE
HILLOCK, WE DON'T KNOW IF THE
MOUSE, WE CERTAINLY DON'T KNOW
IN THE HUMAN.
THE FIRST THING WE NEED TO DO IS
FIND THEM, AND THE LUNG MAP WILL
HELP US DO THAT AND THEN WE CAN
ASK OURSELVES ARE THERE
DIFFERENT FLAVORS OF SQUAMOUS
METAPLACIA.
IT'S A TERM WE THROW AROUND
WITHOUT DEFINING IT.
AND I WOULD ARGUE NOW THAT THE
HILLOCK IS BETTER DEFINED THAN
THE SQUAMOUS METAPLACIAS, MAYBE
THERE ARE A VARIETY OF SQUAMOUS
METAPLACIAS THAT CARE DIFFERENT
IMPLICATIONS.
AND THEN OF COURSE THE MOLECULAR
QUESTIONS, LIKE WHAT IS THE
EPIGENETIC BASIS OF HILLOCK
PLASTICITY AND IT WON'T TAKE YOU
LONG TO REALIZE THAT COULD HAVE
IMP LAYICATIONS FOR
REGENERATION, FOR COPD AND
ASTHMA AND LUNG CANCER AND THEN
THERE'S THIS CURIOSITY OF WHY DO
THE HILLOCK'S NOT HAVE A
STEREOTYPED PATTERN AND THAT OF
COURSE HAS IMPLICATIONS FOR
DISEASE AND I WILL JUST END WITH
THAT AIAN LITTLE BIT EARLY.
BUT WE'VE GOT OURSELVES BACK ON
TIME.
AND IT'S MY PLEASURE TO THEN
INTRODUCE OUR NEXT SPEAKER WHO I
HAD THE WONDERFUL PLEASURE OF
HAVING PASSED THROUGH OUR OWN
LABORATORY QUITE A LONG TIME AGO
BUT NOW TATA IS A PROFESSOR, OR
NO, NO, HE HAS TENURE AT DUKE
AND HAS DONE REMARKABLE WORK IN
THE ALVEOLAR REGION AND INDEED
GLANDS BUT I DON'T THINK WE WILL
TELL YOU ABOUT GLANDS TODAY.
>> GREAT.
THANK YOU, THANKS FOR THE KIND
INTRODUCTION.
AND THANKS TO THE ORGANIZER FOR
HAVING ME HERE, AND ORGANIZING
THIS EXCITING WORKSHOP.
I'M REALLY LOOKING FORWARD TO
LISTEN TO THE REST OF THE TALKS
AS WELL AS THE DISCUSSIONS IN
THE NEXT 2 DAYS.
NEXT PLEASE.
I AM A CELLULAR BIOLOGIST BY
TRAINING BUT THINKING ABOUT THE
LUNG, YOU KNOW 1 IMPORTANT
ASPECT OF THE LUNG THAT WE HAVE
BEEN INTERESTED IN IS THE
ORGANIZATION OF THE LUNG.
YOU KNOW HOW DOES THE LUNG
ACTUALLY ORGANIZE IT'S
SELF-DURING DEVELOPMENT BUT HOW
IT MAINTAINS THE LIFE AS WELL AS
WHEN THERE IS INJURY, CAN IT
REBUILD IT BACK, SO THE REASON
WE GOT INTO THIS IS WE ARE NOW
TALKING ABOUT MANY OF THE
RESPIRATORY DISEASES, 1 KEY
IMPORTANT THING THAT AM CANS UP
IS PHYSIOLOGY.
SO, HERE, WE'RE TALKING ABOUT
FUNCTION, LUNG FUNCTION, AND
THAT'S TIGHTLY ALIGNED WITH THE
STRUCTURES, SO IT'S THE
STRUCTURE FUNCTION RELATIONSHIP
THAT REALLY DEFINES AND IT'S
FUNCTION THROUGHOUT THE LIFE AS
WELL AS YOU KNOW WHEN THERE IS
DAMAGE, OR DIFFERENT DISEASES SO
THAT'S AN ASPECT WE ARE REALLY
EXCITED ABOUT.
NEXT PLEASE EMPLOY AND IN THAT
ASPECT 1 THING WE ARE REALLY
INTERESTED IN IS, WHAT ARE THE
DIFFERENT CELL TYPES AND
DIFFERENT PARTS OF THE LUNG, AND
WHAT ARE THE CELLS THAT HAVE THE
ABILITY TO REPAIR WHEN THERE IS
DAMAGE THAT CAN SUSTAIN TOWARD
THE LIFE.
AND IN THAT PART, I'M MAINLY
GOING TO FOCUS ON THIS DISTAL
PART OF THE LUNG WHICH IS THE
ALVERMEN INFECTED O LIE, SO AS
YOU KNOW, ALVEOLAR TISSUES ARE
TIGHTLY PACKED WITHIN THIS THIN
MEMBRANE WALL, THE ALVEOLAR
RECEPTOR WITH 2 EPITHELIAL
LININGS, AS YOU CAN SEE HERE IN
GREEN, SO WHAT YOU SEE IN GREEN
HERE IS THE ALVEOLAR TYPE 1
CELLULAR MARKER THAT YOU SEE,
THERE ARE 2 LININGS AND BOTH
SITES, SO THERE IS -- THERE ARE
3 DIFFERENT ALVEOLAR SACKS HERE
SEPARATED BY THESE THIN MEMBRANE
AS WELL AND IN THE MITSD WE HAVE
THIS INTERSTITIAL SPACE.
SO HOW ARE THESE CELLS ABLE TO
ORGANIZE THEMSELVES AND MAINTAIN
THROUGHOUT THE LUNG AND WHEN
IT'S TISSUE CULTURE MEDIAD CAN
THEY REBUILD IT BACK?
THESE ARE SOME OF THE QUESTIONS
WE HAVE BEEN ASKING AND I THINK
IT'S A -- FROMAGE ENGINEERING
POINT, IT'S AN ENGINEERING MODEL
AND THE DISEASE BUILT INTO SUCH
THIN, EXTREMELY THIN ALVEOLAR
LINING AND WE KNOW FROM MANY
STUDIES FROM MANY GROUPS THAT
THERE ARE THESE EPIHELIAL CELLS,
THE TYPE 2 CELLS AND THE TYPE 1
CELLS, TYPE 2 CELLS SERVE AS THE
STEM CELLS OF THE ALVERMEN
INFECTED O LIAISON AND THE
INTERSTITIAL FIBROBLASTS AND THE
CAPILLARY IMMUNE SETS AND THE
EXTRA CELLULAR MATRIX BUT MY
TALK IS MAINLY GOING TO TALK
ABOUT THE EPITHELIAL CELLS AND
THE FIBROBLASTS AND WE HAVE BEEN
INTERESTED IN LOOKING AT HOW DO
THESE CELLS RESPOND WHEN THERE
IS DAMAGE, AND HOW DO THEY
INTERACT TWEEZE THEEN 2
DIFFERENT COMPARTMENTS.
NEXT PLEASE.
SO THE REASON WE ASK THAD IS IN
MANY DISEASES, THESE CELLS THE
ALVEOLAR TISSUES ARE UNDERGOING
DISORGANIZATION.
NEXT, PLEASE.
FOR EXAMPLE, IN FIBROSIS IF YOU
LOOK AT IT IN THIS, ALVEOLAR
TISSUES ARE COMPLETELY OCCUPIED
BY THESE MATE RICK PRODUCING MY
O FIBROBLASTS AND MANY OTHER
CELLS INCLUDING IMMUNE CELL ANDS
EPITHELIEL CELLS ARE LOST
EVENTUALLY.
NEXT PLEASE, WHEREAS IN
EMPHYSEMA, THIS IS DEMONSTRATING
LUNG DEC WHERE WE SEE THE LOSS
OF TISSUE, IN OTHER WORDS THERE
IS ALVEOLAR SIMPLIFICATION, SO
WE ARE SIMPLY LOOKING AT THESE 2
OPPOSING PHENOTYPES WHERE THERE
IS A COMPLEX -- INCREASE THE
COMPLEXITY OF THE TISSUE AS SEEN
IN FIBROSIS, THERE ARE
SIMPLIFICATION OF THE TISSUE
LIKE IN EMPHYSEMA AND NOT TO
FORGET, WE ALSO SEE LUNG
CANCERS, ALSO, ALTHOUGH I'M NOT
GOING TO TALK MUCH ABOUT THIS,
BUT WE DO SEE THESE CELLS AT
QUITE DIFFERENT STATES AND
EXPANSION OF THE CELL
POPULATIONS AND THAT'S WHAT THE
CODE OF THE LAB WE'VE BEEN
INTERESTED IN.
NEXT PLEASE.
SO TO CONTINUE, WE'VE BEEN
ASKING THESE QUESTIONS, HOW ARE
THEY ABLE TO GENERATE THESE LARN
TIN TYPE 1 CELLS, WE KNOW THEY
GENERATE BUT HOW DO THEY
GENERATE SUCH A LARGE STRUCTURE
THAT WE SEE IN TYPE 1 CELLS.
NEXT, PLEASE.
TO ADDRESS THIS QUESTION, THERE
ARE SOME TECHNICAL CAVEATS FOR
EXAMPLE, WE CANNOT GET ENOUGH
CELLS FROM NATIVE LUNGS BECAUSE
OF THEIR EXTREME THIN STRUCTURE
OF THESE TYPE 1 CELLS, AND
THAT'S WHERE OUR LAB, AS WELL AS
MANY OTHER LABS HAVE DEVELOPED
ORGANOID MODELS.
IN OUR CASE WE DEVELOPED THESE
CHEMICALLY DEFINED CONDITIONS
WHERE WE TAKE A SINGLE TYPE 2
CELL AND EXPAND THEM INTO 10S
AND THOUSANDS OF CELLS IF NOT
MILLIONS AND ALSO ABLE TO
GENERATE TYPE 1 CELLS USING
THESE ALVEOLAR SPHERE MODEL
SYSTEMS.
TO FORMAL LAB MEMBERS THEY HAVE
TAKEN ON THIS PROJECT AND USED
THESE MODELS TO UNDERSTAND THE
MECHANISMS INVOLVED IN TYPE 2
AND TYPE 1 CONVERSION, NEXT
PLEASE.
AND HERE IS THE SIMPLE --
SIMPLIFIED VIEW OF HOW THE
SYSTEM WORKS SO WE CAN TAKE A
SINGLE TYPE 2 CELL AND
DIFYEPTIATE THEM INTO THE LARGE
TYPE 1 CELL SO THIS IS EXACTLY
THE SAME CELL, IT'S A TIME LAPSE
IMAGE WHERE THE SINGLE CELL
GENERATED THESE LARGE FLAT CELL
OUTSIDE THE BODIES AND USING
MOLECULAR PROFILING AND
DIFFERENT WAYS OF CHARACTERIZING
THESE CULTURES, WE NOW KNOW THEY
ACTUALLY FOLLOW SIMILAR
PATTERNS, MOLECULAR PATTERNS AS
WELL AS MORPHOLOGICAL PATTERNS
WE SEE IN THE HUMAN LUNGS SO
USING THE SYSTEM, NOW WE CAN
LOOK AT MORE REFINELY THE
MOLECULAR PATTERNS.
NEXT PLEASE.
AND WE PERFORM SINGLE CELL
TRANSCRIPTOMIC ANALYSIS AND
FOUND THERE ARE TYPE 2 CELLS,
THERE ARE REPLICATING TYPE 2
CELLS, TYPE 1 CELLS AND THEN WE
CAME ACROSS THIS POPULATION THAT
WE DIDN'T KNOW MUCH ABOUT, NEXT
PLEASE.
AND NEXT.
AND WE FOUND THERE ARE UNIQUE
MARKERS THAT ARE ASSOCIATED WITH
THESE CELL CLUSTERS THAT WE
DIDN'T KNOW BEFORE AND SOME OF
THEM ARE CTGF AND
[INDISCERNIBLE] AND MANY OTHER
MARKERS AND FINALLY WE SEE THIS
TYPE 1 CELL.
SO NOT ONLY WE HAVE THESE
DIFFERENT CELL CLUSTERS AND
MARKERS BUT NOW WE MAP THEM
CHRONOLOGICALLY SO WE KNOW THE
CHRONOLOGICAL EVENTS STARTING
FROM TYPE 2 ALL THE WAY INTO THE
TYPE 1, NEXT PLEASE.
AND NOT ONLY ORGANOIDS, IN
EXVIVO CULTURES BUT WE CLONE THE
SINGLE CELL STAGE IN VIVO UPON
INJURY.
FOR EXAMPLE, IF YOU LABEL
ALVEOLAR TYPE 2 CELLS HERE IN
RED AND FOLLOW UP WITH
BLEOMYCIN, WE SEE THESE UNIQUE
MARKERS THAT COME AS A TRANSIENT
STATE.
AS CAN YOU SEE HERE THEY'RE NOT
PRESENT IN THE STARTING
POPULATION, NEITHER IN THE END
TERMINAL TYPE 1 CELLS BUT THEY
TRANSIENTLY EMERGE DURING INJURY
REPAIR.
NEXT PLEASE.
THE SAME THING CAN BE VISUALIZED
IN THE SCHEMATIC THESE TYPE 2
CELLS GO THROUGH THESE CELL
STATE SO WE LIKE TO CALL THEM
TRANSITIONAL STATE BEFORE THEY
DIFFERENTIATE INTO TYPE 1 CELLS.
NEXT, PLEASE.
AND AN IMPORTANT ASPECT OF THIS
TRANSITIONAL STATUS THAT THE
ACTIVATE MANY SIGNALING PATHWAYS
AND SOME OF THEM ARE VERY
FAMILIAR FROM A DEC POINT OF
VIEW, AS YOU CAN SEE HERE
THERE'S TGFBETTA, P53 SIGNALING
WAGHT PATHWAY, CELLULAR
SENESCENCE, DNA DAMAGE CHECK
POINT, SO NOW WE SEE THERE IS AN
OVERLAP WITH CERTAIN DISEASES
BUT FOR EXAMPLE, IN EMPHYSEMA
AND FIBROSIS, NEXT PLEASE.
SO WE GROW SOME OF THESE IN
LUNGS TO SEE IF WE CAN SEE THE
SIMILAR STATES AND MARKERS.
INDEED WE SEE THEM.
FOR EXAMPLE, IN HEGHTY LUNGS WE
SEE TYPE 2 CELLS BUT IF YOU LOOK
AT THE NONFIBROTIC REGIONS AND
PULMONARY FIBROTIC LUPGS WE
SLEEP APNEA AND OBESITYY
ABUNDANCE OF THESE CELL
DIVISIONS SURROUNDING THE CELLS
THAT OFTEN CALL MY O FIBROBLAST.
SO INDEED THESE TRANSITIONAL
STATES NOT ONLY INJURY REPAIRED
BUT WE SAW AN ARK KIEWMULATION
OF PERSISTENT OF THESE
TRANSITIONAL STATES AND DISEASES
LIKE PULL MANORY FIBROSIS.
NEXT, PLEASE.
NOT ONLY MARKERS BUT ALSO SOME
OF THE SIGNALING ACTIVITIES,
LIKE CELLULAR SENESCENCE, DNA
DAMAGE REPAIR, NEXT PLEASE.
ALSO AS I MENTIONED CELLULAR
SENESCENCE SHOWN HERE WITH THE
STATUS ACTIVATED, WE SEE THESE
IN FIBROTIC LUNGS, BUT NOT IN
HEALTHY CONTROLS.
NEXT PLEASE.
SO HAVING SEEN P53 SIGNALING WE
SIMPLY DISRUPTED P53, USING
CLASSICAL [INDISCERNIBLE] MODELS
AND AND HERE WE FOUND IS A
CONTROL LUNG IN A 3D
VISUALIZATION WHERE YOU ARE
SEEING A SINGULAR ALVEOLAR CUP
FROM 1 VIEW.
THIS IS A CELL TYPE 2 THAT
DIFREBTIATED INTO TYPE 1, NEXT
PLEASE.
WHEREAS IN P53 KNOCK OUT WHAT WE
FOUND IS THESE LINEAGE LEVEL
CELLS DID NOT DIFFERENTIATE INTO
THIS LARGE FLAT TYPE 1 CELL,
INSTEAD THEY KIND OF BALOONED
OUT INDICATING THERE IS
[INDISCERNIBLE] DISREGULATION.
IN ANALOGY 1 COULD TAKE IN A
WAY, SO IN CONTROLS WE SEE THIS
THIN CRUST PIZZA LIKE TYPE 1
CELLS, DERIVED FROM THE TYPE 2.
WHEREAS IN P53 KNOCK OUT SEEEE
THE SOUR DOE LIKE STRUCTURE
INDICATING THAT THERE IS A
STRUCTURAL ORGANIZATION OF THESE
TYPE 2 [INDISCERNIBLE].
NEXT PLEASE, SO JUST TO
SUMMARIZE WHAT I'VE SHOWN YOU SO
FAR UPON INJURY, THESE TYPE 2
CELLS GENERATED THESE UNIQUE
MOLECULARLY UNIQUE TRANSITIONAL
STATE.
NEXT PLEASE.
THAT ARE HIGHLY ENRICHED FOR P53
SIGNALING, DNA DAMAGE AND MANY
OTHER PATHWAYS.
NEXT PLEASE, AND NEXT.
AND NORMALLY, THESE TRANSITIONAL
STATES GO ON TO MAKE THESE LARGE
TYPE 1 CELLS, HOWEVER
UNDERNON[INDISCERNIBLE]
CONDITIONS THESE TYPE 2 CELLS
ARE UNABLE TO DIFFERENT YAILT,
THESE TRANSLATION STATES ARE
UNABLE TO DIFFERENT JAIT INTO
TYPE 1.
APPROXIMATE INSTEAD THEY SPEND
TIME IN THESE TISSUES AND THEY
ARE OFTEN ASSOCIATED WITH THESE
MY O FIEB ROW BLASTS.
SO IN THE AREA WE STUDIED AND
PUBLISHED A FEW YEARS AGO, AND
THERE WERE A FEW PAPERS,
IULOUSED THEM HERE, 1 FROM
[INDISCERNIBLE] GROUP AND
ANOTHER 1 FROM [INDISCERNIBLE]
GROUP IN UK, SIMILARLY
IDENTIFIED THIS TRANSITIONAL
STATE.
AT THAT TIME WE DIDN'T KNOW WHAT
THIS TRANSITIONAL STATE ARE
ACTUALLY DOING.
ALL WE FOUND WAS THERE IS
ASSOCIATION.
AND THAT OPENED UP MORE
QUESTIONS, 1 OF THE QUESTION IS,
ARE THESE TRANSLATIONAL STATES
INVOLVED IN DEVELOPMENT OF
FIBROSIS.
NEXT PLEASE.
@ SAME TIME WE SAW WHEN AND
WHERE THEY EMERGE.
DO WE SEE DIFFERENT INJURY
CONDITIONS THAT IT'S ONLY 1 TYPE
AND THAT TOO, WHAT ARE THE
SENSES IN THE TISSUE.
IN OTHER WORDS IS THERE A LOSS
OF A SINGLE CELL TYPE IN THE
LUNG THAT LEADS TO INDUCTION OF
THESE TRANSITIONAL STATES?
AND THAT'S WHERE WE PROBED
ELIMINATING THESE DIFFERENT CELL
TYPES.
WE TARTED WITH THE TYPE 2 CELLS,
FIBROBLAST ENDOTHELIAL CELLS, I
WILL NOT SHOW ALL THE DATA, NEXT
PLEASE.
TWO PHARMA LAB MEMBERS WHO
CONDUCTED THESE EXPERIMENTS,
NEXT PLEASE, SO WE ELIMINATED
FIBROBLASTS IN CELL TYPES.
IT TURNS OUT THAT ELIMINATION OF
ATP ALONE IS SUFFICIENT TO
INDUCE TRANSITIONAL STATE AND
NOT ONLY THIS EPITHELIAL
TRANSITIONAL STATE BUT WE SEE A
CASCADE OF EVENTS THAT LED TO
WHOLE INJURY REPAIR PROGRAM
THAT'S ACTIVATED IN DIFFERENT
CELL TYPES INCLUDING IN THE
IMMUNE CELLS, FIBROBLASTS AND AS
WELL AS ENDOTHELIAL CELLS.
NEXT PLEASE.
NEXT.
AS I MENTIONED LOSS OF TYPE 1
ALONE INDUCED THESE CASCADE
[INDISCERNIBLE] WHICH CAN BE
SEEN HERE IN THIS HISTOLOGICAL
IMAGE AND THE CONTROL IS SIMPLE
HND WHEREAS AFTER AT1 ABLATION,
WE SEE THESE DENSE FIBROTIC,
SEMIFIBROTIC REGIONS IN THE
GREAT PROCESS.
NEXT PLEASE.
AND WE WANTED TO UNDERSTAND WHAT
ARE THE MOLECULAR EVENTS GOING
ON DURING REPAIR.
SO WE PERFORMED SINGLE
TRANSCRIPT ORDER OF MICRONS
ANALYSIS, THE SINGLE CELL TYPE,
LED TO DEVELOPMENT OF GENERATION
OF THESE CASCADE OF EVENTS
INCLUDING GENERATION OF THE
TRANSITIONAL STAGE.
WE CAN SEE HERE IN A TIME CODE,
NEXT PLEASE, AND WE ALSO --
NEXT, WE ALSO LOOKED AT AMAZING
CHIMAL CELLS, IN THIS CASE WE
INTEGRATE DIFFERENT DATA SETS
FROM OUR OWN MULTIPLE INJURY
MODELS AS WELL AS OTHER DATA
SETS INCLUDING THE INJURY MODEL
FROM THE [INDISCERNIBLE] GROUP.
NEXT PLEASE.
WE FOWBD ESSENTIALLY DIFFERENT
CELL TYPES THAT OTHERS HAVE
DESCRIBED AND SOME UNIQUE
TRANSITIONAL STATES SOME ARE
MARKED BY META1, AND SFRP1 CELLS
AND WE CALL THEM INFLAMMATORY
FIBROBLASTS BASED ON SIGNATURES,
NEXT, PLEASE, AND WE ALSO FOUND
THESE RUNX1 CELLS AND REMODELING
FIBROBLASTS.
NEXT PLEASE.
THE REASON WE CALL THEM -- NEXT
-- THE REMODELING IS BECAUSE IF
WE LOOK AT THE SIGNATURES OF
THESE CELL STATES THESE ARE
TRANSIENT BUT WE SEE THE
FIBROBLAST, WE SEE A LOT OF
THESE COUNTS ASSOCIATE WIDE THE
ECM ASSEM MYELIN, ECM COLLAGEN,
[INDISCERNIBLE] AND VARIOUS
PROTEASES, AS YOU CAN CAN SEE
HERE IN THIS PLOT, THIS
TRANSIENT REMODELING FIBROBLAST,
ACTIVATE A LOT OF THESE ECM
RELATED PROTEINS.
NEXT PLEASE.
AND MORE IMPORTANTLY, SOME OF
THEM HAVE BEEN DESCRIBED AS
ASSOCIATED WITH DIFFERENT
DISEASES THROUGH GWAS STUDIES
INCLUDING THE TGF BETA PROTEIN
AND [INDISCERNIBLE], AND VARIOUS
OTHER ECM RELATED PROTEINS.
NEXT PLEASE.
SO JUST TO SUMMARIZE WHAT WE'VE
SHOWN, I'M NOT GOING TO SHOW ALL
THE MAJORS AND FOLLOW UP DATA
BUT UPON DAMAGE, FIRST WE SEE
IMAGES OF THESE ALVEOLAR
FIBROBLASTS TURNING INTO THESE
SFRP1 +, THEN WE ALSO SEE THE
TRANSITIONAL STATE, POSITIVE
CELLS OR BATPs AND THEN WE SEE
THE RUNX1 SALES DURING THE
INJURY REPAIR PROCESS.
SO WHEN THERE IS A SIMPLE
INJURY, THESE CELLS ARE GOING
THROUGH THE DIFFERENT TRANSITION
STEPS IN THE EPITHELIUM AND THE
FIBROBLAST AND THEY FULLY AREY
PAY, THEY GO BACK TO
[INDISCERNIBLE].
HOWEVER, WE ASKED THE QUESTION,
WHAT IS THE ROLE OF THIS
TRANSITION STATES, WHAT HAPPENS
IF WE DISRUPT THEM, CAN WE
ACTIVATE THEM, CAN WE DEPLETE
THEM THROUGH GENETIC APPROACHES,
OF CELEBRATION APPROACHES.
THAT'S WHAT WE DID.
NEXT PLEASE.
SO HERE IS JUST AN EXAMPLE OF
SCHEMATIC WHEN WE ELIMINATED THE
EPITHELIAL TRANSITION STATE,
THROUGH GENETIC AS WELL AS VIRAL
BASED ABLATION APPROACHES AS
WELL AS FIBROBLASTS, BY DELETING
RUNX1 AND FIBROBLASTS ARE
ELIMINATING THE FIBROBLAST, WHAT
WE FOUND WAS INSTEAD OF REPAIR
THANKSGIVING TISSUE, THIS
TISSUE, ACTUALLY UNDERWENT
SIMPLIFYICATION, MEANING IT'S A
KIND OF A COMPROMISED IN WHICH
NO CELL SET UNDERGOING IMP
LIAISONFICATION INSTEAD OF
REBUILDING THE WHOLE THING, SO
THE WAY WE SEE THIS IS THAT IN
THE ABSENCE OF THESE
TRANSITIONAL STATES, BOTH IN THE
EPITHELIUM AND FIBROBLASTS, THIS
TISSUE IS UNABLE TO REBUILD
BACK, BECAUSE OF LACK OF THESE
STRUCTURAL COMPETENCE INCLUDING
MANY ECM PROTEINS THEY SIMPLY
UNDERGO COMPROMISE, IN OTHER
WORS TISSUE SIMPLIFICATION AS
SEEN IN EMSEEMA.
NEXT PLEASE.
OT OTHER HAND IF WE ACTIVATE
SOME OF THESE CELL STATES,
PERSISTENT, KEEP THEM LONGER
THROUGH EPIGENETIC AND
PHARMAICOLOGGICCAL APPROACHES
NOW WE SEE INCREASE IN TISSUE
COMPLEXITY WHERE DENSE MY O
FIBROBLASTS AND ECM PRODUCED
THAT LOOKS LIKE [INDISCERNIBLE]
FIEBOSEIS, WE PERFORMED
PHYSIOLOGICAL ASSAYS ANDY WOO
SEE SIMILAR OBSTRUCTIVE ON OR
RESTRICTIVE PHYSIOLOGY,AY
REFLECTING THE TOP EMPHYSEMA
PROCESS.
NEXT PLEASE.
SO NOW WE NEED TO UNDERSTAND HOW
THEY CONTROL THESE STATES.
FOR EXAMPLE IN AGING WE SEE MANY
MORE OF THESE ARE IN THE
DISEASES WE SEE, SO WHAT'S
HAPPENING DURING THESE
TRANSITIONS WITHIN EPITHELIUM
AND FIBROBLASTS.
AND ALSO HOW ARE DIFFERENT
SIGNALING PATHWAYS, DNA AND AND
INCREASE IN P53 SIGNALING ARE
MUTATIONS IN P53 SIGNALING,
IMPACT THE TRAJECTORY OF THESE
CELLS AND HOW DO WE RECOVER THEM
WHEN THERE IS ACTIVATION OF
EVIDENT ACTIVATION OF THESE
DIFFERENT PATHWAYS.
THESE 2 DIFFERENT DISEASES
ALTHOUGH FROM A CLINICAL POINT
OF VIEW EMPHYSEMA AND FIBROSIS
AT 2 OPPOSING SITES WE THINK
THAT THESE EMANATE FROM THE
SIMILAR EVENT THAT'S WHY I PUT
MY TITLE EMPHYSEMA AND FIBROSIS
AND 2 WITH THE SAME COIN AND THE
COIN IS THE INJURY REPAIR
PROCESS AND THE PATHWAYS
ASSOCIATED WITH THIS PROCESS.
AND INDEED, IN HUMANS WE KNOW
THAT THESE COMBINED PULMONARY
FIBROSIS AND EMPHYSEMA WHERE WE
CAN ACTUALLY SEE BOTH EVENTS ARE
HAPPENING IN THE SAME LUNGS, IN
SOME CASES IN FACT IN
NEIGHBORING REGIONS IN THIS
CASE, IT SHOWS THERE IS A
SECONDARY NODULE IN 1 SIDE IT'S
EMPHYSEMA, THE OTHER SIDE
FIEBOSEIS, SO WITH THAT, NEXT
PLEASE.
I WOULD LIKE TO THANK ALL MY LAB
MEMBER WHO IS HAVE ACTUALLY DONE
SOME OF THE PRESENT AND PAST
MEMBERS INCLUDING
[INDISCERNIBLE] WHO IS ALSO ON
THIS CALL AND FOR YOUR
ATTENTION.
HAPPY TO TAKE ANY QUESTIONS.
>> THANK YOU TATA FOR A GREAT
TALK.
WE ARE IN THE DISCUSSION PART OF
THE SESSION.
WE CAN GO OVER SOME QUESTIONS IN
THE CHAT BECAUSE I KNOW THAT
THERE ARE SOME GREAT QUESTIONS
THAT MAYBE IT'S BETTER TO
DISCUSS THEM IN PERSON EMPLOY
SO, I KNOW THAT
DR. [INDISCERNIBLE] HAS RAISED
HAND FOR A LONG TIME SO MAYBE WE
CAN GO WITH HIM FIRST.
WE CAN'T HEAR YOU.
>> HEM O.
>> NOW WE HEAR YOU.
>> THANK YOU.
THANK YOU.
THANK YOU ALL FOR -- THIS WAS AN
ABSOLUTELY WONDERFUL SESSION AND
THANK YOU ALL FOR BRINGING SUCH
NEW AND INTERESTING INFORMATION
EMPLOY I HAVE ALREADY A LOT OF
THE ANSWERS ALREADY WITH THE
EXCEPTION OF LAST, TATA, SO I
MAY GET STARTED WITH THAT, WHAT
I HAVE IF I READ WHAT I HAVE
JUST WRITTEN IT IS REMARKABLE
HOW MUCH THESE DISEASEITATES
NEAR DEVELOPMENT AND IN FACT,
BOTH IN HUMAN AND MICE, IN FACT
THIS, IS A PUBLICATION THAT IT'S
SOON TO COME FROM OUR LAB IN
CELL AND WHAT I WOULD LIKE TO
LEARN TATA, IS PROBABLY A
QUESTION THAT WE HAVE AND IT
APPEARS WHETHER THIS IS -- IS
THERE A LINK BETWEEN A
CHRONOLOGICAL LINK BETWEEN THE
APPEARANCE OF THESE MARKERS, FOR
EXAMPLE IN WHAT YOU HAVE
DESCRIBED IN THE FIBROSIS AND
ALSO, IS THERE ANY EVIDENCE THAT
WE CAN THINK THAT ALL OF THESE
MARKERS ARE FUNCTIONAL IN SOME
WAYS AND THERE'S OF COURSE THE
WORK FROM RACHEL [INDISCERNIBLE]
WHO TALKS ABOUT THE KERATIN 8,
YOU HAVE VERY INTERESTING
OBSERVATION ABOUT THE P53, BUT
THERE'S SO MANY OF THESE MARKERS
AND SO CONSERVATIVE, PLEASE ANY
THOUGHTS ABOUT THAT?
>> YEAH, ABSOLUTELY.
SO THE TRANSITION STATES IN
DISEASE, SO [INDISCERNIBLE] MOST
OF THE PROFILINGS AND ANALYSIS
THAT HAS BEEN DONE IN THE FIELD,
INCLUDING OUR OWN FROM HUMAN
TISSUES POINT OF VIEW, CAN'T
COME FROM LATE STAINLS, EVEN
THEN WE CAN ACTUALLY SEE
GRADIENTS OF [INDISCERNIBLE].
SO THERE IS A REGION THAT'S
PROGRESSING AND WELL IS ALREADY
A DENSE FIBROTIC REGION,
BRONCHIALIZED REGION FOR EXAMPLE
IN FIBROSIS SO WE SEE THESE
EVENTS, THE DIFFERENT TYPES OF
[INDISCERNIBLE] AND STATES
ASSOCIATED MARKERS WAS THE
EPITHELIUM AND THE MESSENTERIEN
KINE IN THESE PROGRESSIVE ZONES
SO IN A WAY THE PROGRESSIVE
ZONES ACTUALLY GIVING DOES --
VIEW THAT, YOU KNOW HOW -- WHAT
ARE THE CHRONOLOGICAL EVENTS
THAT ARE HAPPENING IN THE
DISEASE.
>> GREAT.
THANK YOU EMPLOY THANK YOU.
I WILL LEAVE YOU WITH THE OTHER
QUESTIONS OTHERWISE, I
UNDERSTAND TO THE AUDIENCE.
>> THAT IS A GOOD QUESTION, IF I
MAY READ OUT FROM CHRIS MOLENA,
ASKING ABOUT THE INJURY MODELS
THINKING THE RISES AND FALL OFF
OF RTP1 POSITIVE FIBROBLASTS
REGULATES AT281 TRANS8 HOURS,
INDEED ACTUALLY SO THIS IS
UNPUBLISHED DATA BUT WHAT WE
FOUND IS THESE SFP1 FIBROBLASTS
ARE THE FIRST TO EMERGE AND THE
WAY WE KNOW THAT IS, YOU KNOW IN
HUMAN IT'S HARD TO KNOW WHICH 1
COMES FIRST, WHICH 1 BUT THIS
ANIMAL MODELS WE CAN POST THE
INJURY AND THAT'S WHERE I LIKE
THE ABLATION MODEL, WE CAN
ABLATE THE TYPE 1 CELLS AND THEN
FOLLOW CHRONOLOGICALLY AND
THAT'S WHERE WE FOUND SFRP1 +
FIRST.
AND IF YOU LOOK AT THE NATURES,
THEY PRODUCE MANY INFLAMMATORY
CYTOKINES THAT RECRUIT
MACROPHAGES, SOME OF THE IMMUNE
POPULATIONS AS WELL AS THAT ALSO
STIMULATE THE NEIGHBORING TYPE 2
CELLS TO TRANSITION TO THE PATH
POPULATIONS, THE TRANSLATIONAL
STATES.
SO WE THINK THAT SFRP1 +
POPULATION IS ACTUALLY INDUCING
TYPE 2 CELLS TO INITIATE THE
EPITHELIAL AREA.
I HOPE THAT ADDRESS YOUR
QUESTION CORRECTLY.
>> I'D LIKE TO ADD A COUPLE --
LIKE 1 COMMENT, I JUST WANT TO
SHARE THE OBSERVATION THAT WE
HAD, I KNOW THAT TATA, JUST
MENTIONED THAT IN HE THINKS THAT
SORT OF LIKE A TRANSITIONAL
FIBROBLAST COULD AFFECT OR
INITIATE THE AT2 OR AT1 INJURY,
AT LEAST IN MY MODEL, THE WORK
THAT I JUST SHARED IN
DEVELOPMENTAL MODEL, WE
BASICALLY PERTUSHED EPITHELIUM
AND WE SAW THE EMERGENCE OF
THOSE SFRP 1 POSITIVE AND THE
RUNX1 POSITIVE FIBROBLAST
MENTIONED THERE SO AT LEAST IN
MY MODEL, THE CHANGES IN THE
EPITHELIUM INDUCE THE CHANGES IN
FIBROBLAST.
>> YEAH, SO THOSE ARE BELIEVE
THE GENETIC MODELS, WHERE ARE
YOU ACTIVATING AND DELETING PART
OF THE GENE IN EPITHELIAL SALES.
SOPHISTICATEDY WOO DID COMPARE
MDMKNOCK OUT WHICH INDUCES P53,
AND THAT'S WHAT WE DID AND THAT
MODEL WE SEE.
AND IF WE APPROXIMATELY THIS
CHRONOLOGICAL ORDER WHAT WE SEE
IS, WE SEE THIS, THE SFRPT1 +,
AND THEN BECOMES RUNX1 POSITIVE
BUT THE QUESTION YOU BRING UP
AND WHO INDUCES WHO?
>> YES, YES.
>> IT CAN HAPPEN IN MULTIPLE
WAYINGS BUT IN A SIMPLE MODEL WE
THINK IT'S SFRP1 THAT INITIATES
TAKEN--THEY THE TYPE 2 CELLS.
THAT'S THE TRIGGER.
>> I SEE, I SEE.
>> BECAUSE YOU SEE LIKE VERY
INTERESTING PLOT INCREASE IN THE
IMMUNE CELL, IN THE COPD
PATIENTS IN PARTICULAR,
T-RESIDENTIAL MEMORY TISSUING
CELLS, I WONDER WHETHER YOU SEE
THE PREVIOUS [INDISCERNIBLE] AND
THE CD8 T-CELL INCREASE THE COPD
IN THE PATIENT.
DO YOU SEE ANY MARKER FOR THAT?
>> FOR THAT IN THE BRONCHIAL
ABLIT RANS.
IN COPD I DON'T HAVE IA LOT OF
PATIENT WHERE IS I COLLECTED
PARALLEL IMMUNE ENRICHED SAMPLES
WHICH IS HOW I APPROACHED THE
MANY OF THE RARE DISEASES
INCLUDING BRONCHIALITEIS OBLIT
RANS SO I CAN'T.
WE DO HAVE SOME EMPUNE CELLS IN
OUR COPD DATA SETS BUT WE
SPECIFICALLY DEPLETE 6 CD45 IN
ORDER TO ENRICH FOR THE
EPITHELIUM.
IN THE RARE DISEASES LIKE PH OS,
WE RUN THOSE IN PARALLEL SO WE
HAVE 2 DATA SETS WE CAN USE SO I
CAN'T SAY THAT I'VE LOOKED
EMPLOY.
>> THANK YOU.
NTHERE IS ANOTHER QUESTION FROM
MAURICEIO IN THE CHAT: DO YOU
THINK THIS IS ALSO THE CASE IN
OTHER INJURY MODELS LIKE
INFLUENZA VIRUS.
INDEED, ACTUALLY, YOU SEE THE
SIMILAR TRANSITION STATES THAT
ARE MATCH NOTHING DIFFERENT
INJURY MODELS.
WE HAVE TESTED LPS FLU MODEL,
HUMAN ACTIVE MODEL, ABLATION
MODELS IN DIFFERENT.
>> WE HAVE ANOTHER QUESTION
SAYING ANY COMMENTS ON THE ROLE
OF LGR5 CELLS IN ANY OF THESE
INJURY MODELS.
>> MAYBE I CAN START, SO LGR5
POSITIVE CELLS, WHAT WE HAVE
SEEN, THESE ARE PRESENT IN THE
PERIBRONCHIAL REGION, NOT IN THE
SPACE, AT LEAST DATA WE HAVE
SEEN IN HUMAN TISSUES.
AND I AM NOT SURE, IN MOUSE
MODELS OF ANYONE HAS THOSE
SPECIFIC LGR 5 USING THE LGR 5
BASED TOOLS BUT THERE ARE OTHER
MARKERS ASSOCIATED WITH THE
SIMILAR CELL TYPES AND INDEED,
THEY DO HAVE A ROLE IN
EPITHELIAL REPAIR IN THE
BRONCHIALS.
>> WE HAVE A QUESTION FROM
DR. SUSAN [INDISCERNIBLE] IS
THAT THE ABILITY OF AT2 CELLS
GIVE RISE TO A TRANSITIONAL CELL
IN ORDER TO ULTIMATELY BECOME
AT1 CELLS, IS A GREAT ADVANCE IN
UNDERSTANDING THE REGENERATION
OF THE [INDISCERNIBLE] OR
EPITHELIUM INJURY, CAN ANY OF
THE SPEAKERS SPEAK TO HOW THEIR
RESEARCH PROGRAMS ARE ADDRESSING
THE RESILIENCE AS APPROXIMATE
PERTAINS TO AT2 SELF-RENEWAL?
>> DR. HAN, MAYBE YOU WANT TO
TAKE THAT?
>> YES, IT COULD BE MORE OF AN
ABILITY TO PROLIFERATE FOR THE
AT2 CELLS, I WILL TRANSLATE IT
THAT WAY, I MEAN OBVIOUSLY THE
RESEARCH THAT I SHOWED SO FAR IS
MORE OF THE CELL FATE, AND
DIFFERENTIATION RATHER THAN
PROLIFERATION.
WE HAVE SOME UNPUBLISHED DATA TO
SUGGEST THAT 1 OF THE ISR
PATHWAYS ESPECIALLY FOR THE ATF
4, MAY BE INVOLVED IN THE CAUSAL
PATHWAY FOR CELL PROLIFERATION
AS LIKE AN AT2 STEM CELL
PROLIFERATION.
BUT, THAT'S ALL I CAN SAY, MAYBE
DR. [INDISCERNIBLE] CAN FILL IT
IN.
>> SURE, SURE.
THAT'S AGAIN WHERE WE TARTED
USING ABLATION MODEL.
WE PUBLISHED SOME OF THE WORK SO
IF WE ABLATE TYPE 1 CELL, TYPE 2
CELL, THE FIRST PREFERENCE IS TO
DIFFERENTIATE.
SO IF WE JUST LOOK AT THE EVENTS
OF WHAT ARE THE [INDISCERNIBLE]
OF TYPE 2 CELLS, FIRST BEGINS
DIFFERENTIATION.
IT GENERATES THE TYPE 1 CELLS
AND THEN, THE REMAINING TYPE 2
CELLS SURVEYED IN THE
NEIGHBORING ALVERMEN
NEIGHBORING ALVOELI, AND WE HAVE
2 LENGTHS OF EVIDENCE, 1 USING
CLONAL LABELING EXPERIENCE WHERE
WE CAN LABEL CLONEALLY TYPE 2
CELLS AND ELIMINATE SOME OF THEM
AND THESE 2 MAKE ONLY TYPE 2
CELLS.
AND THIS HAS BEEN SHOWN BY
ANOTHER PAPER RECENTLY BY IT WAS
A COLLABORATION WITH MAURICEIO
[INDISCERNIBLE] AND
[INDISCERNIBLE] USING LIVE
IMAGING APPROACHES WHERE WE CAN
SEE THE SAME THING.
TYPE 2 CELLS FROM 1 ALVEOLAR
COMPARTMENT MIGRATE INTO OTHER
COMPARTMENTS, THEY CAN
ACTUALLYY PENETRATE THROUGH THE
WALL, THEY CAN GO TO THE CONE TO
REACH THE NEIGHBORING ALVEOLAR
SACKS AND THIS IS WHICH THROUGH
THEY ARE TYPE 2 CELLS RECRUITED.
HOWEVER, IN MY MIND WHAT WE
DON'T KNOW IS, WHAT HELPS THE
TYPE 2 CELL.
THERE IS ANOTHER TYPE 2 CELL
MISSING IN NEIGHBORING
COMPARTMENT.
WE DON'T KNOW WHAT ARE THE TYPES
OF SIGNALS THAT THE SECRETERS
FACTORS THAT ARE COMMUNICATING
THROUGH THESE CELLS OR
NEIGHBORING CELLS LIKE
FIBROBLASTSER OTHER CELLS
COMMUNICATING, THERE ARE
DIFFERENT MECH NICHES, WE DON'T
KNOW THAT.
SO SEEMS LIKE THE TYPE 2 CELLS,
AND DIFFERENTIATION ARE TIGHTLY
CONTROLLED, AT LEAST BASED ON
SOME OF THESE ABLATION
EXPERIMENTS WE'VE SEEN.
DO YOU KNOW WHEN TO REPLICATE,
THEY KNOW WHEN TO DIFFERENTIATE
AND THEY HAVE A FIRST PREFERENCE
IN A BROADER INJURY MODEL, THEY
MUST DEFERENTIATE BEFORE THE
REPLICATION NETWORK QUICK
QUESTION.
SO YOU THINK THAT WITH ABLATION,
THE TYPE 2 CELL INITTIAIATES
DIFFERENTIATION FIRST AND THEN
TRY TO SELF-RENEW LATER?
>> THAT'S WHAT WE SEE, AT LEAST
IN TYPE 1 CELL ABLATION MODEL.
BECAUSE IN OTHER INJURY MODELS
IT'S HARD TO GUESS THAT.
>> I MEAN THAT'S WHAT THE
[INDISCERNIBLE] PUBLISHED A
COUPLE YEARS AGO.
ESSENTIA WILY THAT'S WHAT HE'S
SEEN TOO, BUT THEN THINKING
PHILOSOPHICALLY WHY WOULD BODY
WANT TO DIFFERENTIATE BEFORE
PROLIFERATE TO RE7 THEIR STEM
CELL?
PHILOSOPHICALLY DO YOU ANY
THOUGHTS ON THAT?
>> I THINK CONNECTED TO
PHYSIOLOGY.
WHAT IS THE MOST IMPORTANT CELL
TYPE IN THE LUNG IN MY VIEW IT'S
THE TYPE 1 CELL BECAUSE THAT'S
THE FUNCTIONAL CELL THAT'S
PARTICIPATING IN BREATH
EXCHANGE.
THAT'S THE FIRST PLACE IT NEEDS
TO PASS THROUGH BEFORE IT GETS
TO THE ENDOTHELIUM.
IT NEEDS TO MAKE THAT GAS ICS
CHANGING CELL.
-- GAS EXCHANGING CELL.
>> DR. JAY WAS GOING TO SAY
SOMETHING.
>> OH NO, SIRRA LYNN ASKED A
VERY THOUGHTFUL QUESTION, IT
ISN'T IN DETAIL BUT RATHER HAS
CONCEPTUAL IMPLICATIONS FOR DEC
AND SHED SOME INJURIES ARE ACUTE
WHILE THE CHRONIC DISEASE
DEVELOPMENT PROCESS STIEPS TAKES
DECADESMENT ARE THERE SHORT
LIVED VERSUS LONG LASTING
TRANSITIONAL CELL STATES WHERE
THE LATTER BECOME A STABLE
PATHOLOGIC CELL TYPE?
SO THAT'S A VERY INTRIGUING
QUESTION AND FOR EXAMPLE, 1
PLACE WHERE THAT MIGHT COME UP
IS IN THE IDEA OF FIELD
CANCERRIZATION.
WHERE HAVE YOU A SUBCLINICAL
CLONE THAT THEN EXPANDS AND YOU
WOULDN'T EVEN KNOW THAT IT'S
INCREASED AND THERE WOULD BE THE
SIMPLE MINDED EXPLANATION OF
ONCOGENIC STRESS THAT WOULD
CAUSE THAT SO JUST A COMPETITIVE
CELL COMPETITION MODEL, WHERE
YOU WOULD GET A BIG FIELD THAT
HAD LET'S SAY P53 OR KRASE
MODEL, AND THEN THAT FIELD WOULD
THEN BE SUBJECT TO MORE
GENOTOXIC STRESS, SO I THINK
SARAH THAT'S 1 WAY YOU COULD
IMAGINE IT, BUT NOW WE'RE
LOOKINGA THE A DIVERSITY OF
DIFFERENT CELL TYPES SO IT COULD
BE FOR EXAMPLE, IN MY CASE, THE
HILLOCK BASAL COULD EXPAND AND
THEN IT COULD ACTUALLY ALSO
BECOME PLASTIC.
BUT THAT DOESN'T MEAN THAT THAT
CELL THAT'S BECOME PLASTIC
DOESN'T RETAIN AN EPIGENETIC
MEMORY OF ITS HILLOCK STATE.
IN WHICH CASE THE ENTIRE
EPITHEMUAL MAY HAVE A
PREDISPOSITION TO BECOMING
SQUAMOUS LATER AND BECOMING
INJURY RESISTANT.
AS I MENTIONED VERY BRIEFLY THAT
SAM [INDISCERNIBLE] AND WHO IS
THE FELLOW AT UC -- PETER
CAMPBELL HAD PREVIOUSLY
DESCRIBED THE CELL TYPE THAT
EMERGED AFTER WHOSE
PROPORTIONALITY INCREASED AFTER
THE CESSATION OF SMOKING.
SO I THINK THAT'S EXACTLY WHAT
YOU'RE KIND OF TALKING ABOUT,
AND I THINK THAT'S PROBABLY A
HILLOCK LIKE CELL OR SOME OTHER
CELL THAT THEN HAS A SELECTIVE
ADVANTAGE LATER, BUT THEN THOSE
SAME CELLS WILL BE ANDROGENIC,
BUT AT SOME POINT, THE RESILIENT
CELL ITSELF WHICH EXPANDED
DURING THE COURSE OF INJURY THEN
BECOME SUBJECT TO PATHOLOGY.
SO I THINK IT'S VERY, YOU KNOW
IF THAT'S THE ONLY CELL LEFT,
THEN IT'S GOING TO BECOME A
CANCER.
BUT I THINK IT'S A VERY
THOUGHTFUL QUESTION, AND I THINK
EACH DIFFERENT KIND OF INJURY
PARADIGM, IT'S GOING TO HAVE A
DIFFERENT ANSWER.
AND I LIKE VERY MUCH THE WAY YOU
PHRASED LIKE, THE ACUTE RESPONSE
TO INJURY, VERSUS THE CHRONIC,
BECAUSE IT'S A DISTINCTION WE
SEE IN THE CLINIC ALL THE TIME
AND VERY INTERESTING.
>> JAY, I HAVE A FOLLOW UP
QUESTION.
I HAD A FOLLOW UP QUESTION TO
THAT ACTUALLY.
>> SERE A THINK SARAH'S TRYING
TO SAY SOMETHING.
>> SORRY, SO REAL QUICKLY, I
THINK MY QUESTION WAS TRIGGERED
BY TATA, SAYING INJURY RESOLVED,
TRANSITIONAL CELLS WENT BACK TO
NORMAL SO THIS IS RELATED AT
WHAT POINT THIS IS REVERSIBLE
VERSUS NOT?
I THINK FOR LOSS OF THE CHRONIC
DISEASE FIBROTIC OR EMPHYSEMA,
WE SEE THAT IS TERMINALLY STAGED
SO THAT MEANS THEY'RE NOT
REVERSIBLE ANYMORE.
SO, I THINK THIS IS A MILLION
DOLLAR QUESTION WHETHER WE CAN
PUSH THE REVERSIBLE ACTION OF
THE TRANSITIONAL CELL TYPE, SO I
WILL STOP HERE.
THANK YOU BOTH EMPLOY REALLY
NICE TO SEE EVERYONE.
>> JOSE DID YOU HAVE A QUESTION.
>> YEAH, JUST TO KIND OF FOLLOW
UP ON THAT.
MAYBE A PROVOCATIVE QUESTION,
ARE HILLOCK'S REQUIRED?
LIKE DO YOU NEED THEM IN A
NORMAL PHYSIOLOGICAL STATE?
AND HOW EARLY DO YOU SEE THEM,
DO YOU THINK THEY ARE THE RESULT
OF MINOR INJURIES THAT OCCUR,
ALMOST KIND OF MISTAKES THAT
LEAD TO THAT PATCH WORK EMERGE
FROM THAT OR IS IT MERELY A
HOMEOSTATIC STRUCTURE?
>> SO THESE ARE FASCINATING
QUESTIONS, YOU KNOW AND I DON'T
HAVE ALL THE ANSWERS, TO YOUR
FIRST QUESTION ARE HILLOCKS
NECESSARY, IT REQUIRES AN
ABLATION AND IT REQUIRES THE
ABLATION TO BE SO EXTREME THAT
YOU KNOW YOU WOULD NEARLY NEED A
HUNDRED PERCENT EFFICIENCY.
SO WE CAN'T ANSWER THAT
QUESTION.
AND IT WOULD BECOME
QUANTITATIVE.
YOU KNOW?
BECOME A QUANTITATIVE RESCUE OF
A REGENERATION AND OUR INJURY
MODELS IN MOUSE ARE SO PRIMITIVE
THAT THEY CAN NEVER BE
STANDARDIZED SUFFICIENTLY TO GET
LIKE, YOU KNOW IF YOU THINK
ABOUT HDM OR YOU THINK ABOUT
SO2, YOU WILL NOT PULL OUT GREAT
ANSWERS FOR THAT.
SO WE WILL HAVE TO DO DIFFERENT
KINDS OF EXPERIMENTS WHERE WE
MARK CELLS AND FOLLOW THEIR
LINEAGE AND SEE LIKE, HOW MUCH
THEY'RE ABLE TO COMPETE WITH
THEIR NEIGHBORS.
NOW THE SECOND QUESTION IS EVEN
MORE INTRIGUING JOSE AND I
THINK, YOU KNOW I'M NOT GOING TO
BE ABLE TO GIVE YOU A SIMPLE
ANSWER, BECAUSE IT KEEPS
CHANGING.
BUT I WOULD HAVE TOLD YOU BEFORE
THAT THE HILLOCK PATTERN IS
STABILIZED AT ABOUT 8 WEEKS.
BUT IT TURNS OUT THAT IF YOU
LOOK VERY SUBLY THEN WE CAN GO
BACK TO 2 WEEKS WHERE THERE'S A
HILLOCK PATTERN AND BUT THEY'RE
MUCH SMALLER HILLOCKS.
AND SO, AN UNDERLYING QUESTION
BECOMES IS THERE THE COMPETENCY
TO FORM HILLOCKS THAT IS
ACTUALLY STEREO TYPICALLY
DISTRIBUTED THAT, IS TO SAY VERY
RARE CELLS WHERE IT WOULD BE
LIKE A NORMAL
DEVELOPMENTALLENTITY WHERE YOU
JUST HAVE SCATTERED HILLOCKS
THAT WERE BILATERALLY SYMMETRIC
ALONG THE ACIS AND SOMETHING
HAPPENS AND YOU CAN HAVE A
MASSIVE EXPANSION OF THAT CELL
TYPE, LIKE JOHN [INDISCERNIBLE]
WHO IS ON THE TALK PREVIOUSLY
SUGGESTED THERE MAY BE SOMETHING
LIKE THIS AT THE ESOPHAGEAL
GHASTIC POUNDRY SO THAT REMAINS
POSSIBLE.
AND THEN TO THE THIRD COMPONENT.
YOU ASKED A VERY COMPLICATED
QUESTION, ALL OF IT INTERESTING.
I THINK WHY ARE THEY THIS WAY?
AND IT MIGHT BE AS SIMPLE AS THE
MOUSE HOLDING ITS NECK IN A
CERTAIN DIRECTION AND THE
ESOPHAGUS PRESSING UP AGAINST
THE TRACHEA IN A CERTAIN WAY
THAT WOULD BE DIFFERENT IN
DIFFERENT PEOPLE, DIFFERENT MICE
FOR THAT MATTER.
AND I DO THINK AS A QUESTION,
WELLING TON OR [INDISCERNIBLE]
ASKED THAT THESE THINGS ARE
CAUSED TO OUTGROW BY PHYSICAL
TRANSDUCTION, BY SOME KIND OF
MECHANO TRANSDUCTION PATHWAY AND
I THINK THE BEST WAY TO THINK OF
THEM METAPHORICALLY IS THE
CALLOUSES ON YOUR HAND.
THERE'S THE CAPACITY TO FORM
CALLOUSES AND THEY'RE NOT
STEREOTYPE, YOU KNOW IT DEPENDS
ON HOW YOU HOLD THE OAR AS YOUR
ROWING, BUT EVERY DAY WITH THE
WEAR AND TEAR, YOUR HAND IS NOT
AS SMOOTH AS A BABY, SO THROUGH
WEAR AND TEAR YOU DEVELOP
POPULATIONS THAT ARE NOT STEREO
TYPICALLY ARRANGED BUT ARE
FUNCTIONALLY USEFUL.
DOES THAT ANSWER YOUR QUESTION
JOSE?
>> YES, EXCELLENT.
THANK YOU.
NTHERE IS A QUESTION FROM
[INDISCERNIBLE] CHEN, MAYBE THIS
IS RELATED TO MARIAS WHAT ARE
YOUR THOUGHTS ON THE TERMINAL
CELL CONTRIBUTION TO THE
ALVEOLAR TRANSITIONAL STAGE LIKE
IN IBF?
>> YEAH, SO I THINK IT'S
POSSIBLE.
I THINK THERE'S THIS IDEA OF THE
[INDISCERNIBLE] POPULATION IN
IBF THAT MULTIPLE GROUPS HAVE
SHOWN IN SINGLE CELL SEQUENCING,
SO PEOPLE WHO ARE HERE, WHICH I
THINK YOU KNOW MIGHT BE A
DISTINCT POPULATION POTENTIALLY
FROM THE TRANSITIONAL CELL
STATES, AND MAY BE RELATED BUT
MAY ALSO BE DIFFERENT AND IF YOU
USE SORT OF BIOINFORMATIC
ANALYSIS, AND IT LOOKS LIKE
THOSE COULD BE RELATED TO THE
AIRWAY COUNTERPARTS AT LEAST IN
HUMAN CELLS.
BUT TESTING THAT DIRECTLY WILL
REQUIRE EITHER ORGANOID MODELING
AND UNDERSTANDING THE SIGNALS
THAT GO INTO THAT, OR SOME OTHER
SORT OF ANIMAL MODEL WITH
LINEAGE TRACING LIKE IN THE
FERRET MODEL.
>> THERE WAS A PAPER FROM
[INDISCERNIBLE] GROUP WHO HAD
THE [INDISCERNIBLE] LINEAR MODEL
AND THEY FOUND SOX 2 POSITIVE
CELLS, ALTHOUGH THEY DIDN'T GET
TO THE DETAILS, WHICH 1 EXACTLY
BUT THEY CAN ALSO GENERATE THIS
TRANSITIONAL STATE WHICH THEN
EEIVETTUALLY BECOME TYPE 1 CELLS
ALTHOUGH IT'S TO A LOWER DEGREE,
NOT MANY CELLS.
>> AND THE QUESTION IS ARE THOSE
THE AIRWAY CELLS THAT CONTRIBUTE
TO THE ALVEOLAR CELLS THAT GO ON
TO NORMAL DIFFERENTIATION AND
THE CELLS IN THE HUMAN
POPULATION THE SAME OR
DIFFERENT?
>> YEAH, YEAH, THERE IS A BIG
DIFFERENCE BETWEEN MOUSE AND
MUSEUM MAN ACTUALLY, THESE
TRANSITIONAL STATES BECAUSE THE
HUMAN EVIDENT BASED CELLS AS THE
NAME SUGGESTS THEY EXPRESS THESE
BASE WILL CHARACTERISTIC MARKERS
BUT IN MICE WE HAVEN'T SEEN ANY
OF THE MODELS WE TESTED.
>> YEAH.
>> TATA, I HAVE A QUESTION,
FOLLOW UP WHY YOU DESCRIEWB
EARLY, WHEN YOU HAVE LIKE TYPE 2
CELL TRENDS ALVEOLAR INTO THE
OTHER SPACE FOR REPAIR, THAT
PARTICULAR TYPE 2 CELL MAY BE
ALREADY IN THE TRANSITION STAGE
AND THEN TARTED REPAIR PROCESS
WHILE MAINTAINING TYPE 2 CELL
FEATURE?
>> YEAH SO THAT'S ACTUALLY
REALLY INTERESTING PART.
THE PRECISE QUESTION WE TESTED
USING THIS ABLATION MODEL, IF
YOU ELIMINATE TYPE 2 CELLS, THEY
MAY GREAT THE REPLICATE, THEY
MIGRATE AND MAINTAIN AS ONLY
TYPE 2, WE DON'T SEE ANY
[INDISCERNIBLE] TO TYPE 1.
SO THEY KNOW XAKLY WHEN TO
RESPOND TO EITHER REPLICATE,
MIGRATE, VERSUS DEFERENTIATE.
YEAH, HOWEVER, IF YOU DO THE
SAME THING IN [INDISCERNIBLE]
MICE IN INJURY MODEL FOR
EXAMPLE, THEY CAN MIGRATE AND
REPAIR GENERATE TYPE 1 CELL
SPACE, SO IT'S ALL ABOUT THE
NEED.
>> THANK YOU.
>> JAY, YOU HAVE A QUESTION FROM
MAURICIO, ARE THESE STATES
SIMILAR TO TRANSIENT
POPULATIONS?
>> IN AS MUCH AS PATS ARE
FUNCTIONAL NORMAL INTERMEDIATES,
I THINK THAT WOULD BE AN
APPROPRIATE COMPARISON.
THEY MAY HAVE SOME HOMEOSTATIC
CONSTANT FUNCTION THOUGH WHICH
WOULD BE DIFFERENT, SO IN OTHER
WORDS, YOU COULD HAVE A
TRANSIENT STATE THAT IS NOT
FUNCTIONAL, ALTHOUGH THERE'S
BEEN TALK THAT YOU KNOW THE
TRAPPED STATE IN DISEASE WILL
HAVE FUNCTIONAL IMPLICATIONS.
THE HILLOCK AS FAR AS WE CAN
TELL DOESN'T HAVE ANY DETAILS
RIMENTAL STATE FUNCTION AND WE
LOOKEDDA THE THAT AND 1 OF MY
POST DOCTORAL FELLOWS CANNED
CANNED --
[INDISCERNIBLE] HAS PUBLISHED
THAT THERE ARE PATCHES IN THE
BACK OF THE HUMAN AIRWAY THAT
WERE BRAHMAATICALLY SIMILAR TO
HILLOCKS BUT WHEN SHE CULTURES
THOSE IN A BUNCH OF SIGNALING
CASCADES SHE SEES A WEAKENED
BARRIER SO BECAUSE OF THE
DISTRIBUTION OF THESE CELLS IS
SO SIMILAR TO THAT OF HILLOCKS,
IT'S POSSIBLE THAT THEY'RE
DISEASED HILLOCKS SO MAURICIO,
I'M HAVING A HARD TIME
ARTICCULING A SPECIFIC ANSWER TO
YOUR QUESTION BUT I GUESS WHAT I
WOULD SAY IS MAYBE YOUR ANALOGY
IS ACTUALLY QUITE A GOOD 1.
BECAUSE THE HILLOCKS MAY BE
SUPER TRANSITION STATE AT BASE
LINE BUT I SUPPOSE WHAT I'M
SEEING IS THAT THIS GROUP OF
EXCELLENT YOUNG INVESTIGATORS
HAS TAKEN A DISEASE AND SHOWN
HOW THAT TRANSITION STATE MAY BE
AN ABERRANT PATHIO LOGICALLY
SIGNIFICANT ENTITY BUT WE
HAVEN'T YET DONE THAT WITH THE
HILLOCK AND I THINK THAT IS WHAT
WE NEED TO DO SO I THINK WE NEED
TO LOOK AT COPD AND ASTHMA AND
ASK FOR THESE ABERRANT HILLOCKS
THAT COULD BE CAUSING PROBLEMS?
SO THAT -- AT FIRST I WAS
CONFUSE BY YOUR QUESTION BUT I
THINK IT'S A GREAT QUESTION.
ONE I WILL THINK MORE ABOUT.
>> THERE IS A QUESTION FROM
DAVID [INDISCERNIBLE].
HE IS ASKING ABOUT GIVEN THE
EXTENSIVE PLASTICITY OF THESE
DISTAL LUNG CELLS, IT IS HARD TO
UNDERSTAND WHERE RESILIENCE
FALLS AND LUNG DISEASE DEVELOPS,
FROM MY SOMEWHAT [INDISCERNIBLE]
PERSPECTIVE, I WOULD ASK WHERE
DO THESE CELL MECHANISMS FAIL.
WHERE ARE WEAK LINKS IN CELL
PLASTICITY?
MARIA?
OR SEUNG, DO I WANT TO TAKE ON
THAT?
>> YEAH, I DON'T THINK IT'S NOT
NAIVE AT ALL.
I THINK THAT -- THE KEY NOTE
RIGHT WHICH IS THE WHYED THAT IF
WE REALLY WANTED TO DRILL DOWN
ON THAT HUMAN SPECIFIC SYSTEM,
WE EITHER HAVE TO DEVELOP
LONGITUDINAL COHORTS SO WE CAN
UNDERSTAND HOW THE CELL
SIGNALING CHANGES OVER THESE
DEVELOPMENTS OR WE HAVE TO FIND
MODELS AND THE MODELS MAY BE
DIFFERENT FOR EACH CELL TYPE
INTERACTION, RIGHT?
THE BEST CELL SYSTEM TO STUDY IN
THE AT1 OR 2 TRANSITION IS MAY
NOT BE THE BEST MODEL SYSTEM
THAT WE STUDY THE DEVELOPMENT OF
THE BASAL CELL IF IT COMES FROM
AN AIRWAY BASAL CELL OR
SECRETATTORY, SO OW HE USE OUR
MODELS OR LAUNCH LONGITUDINAL
COHORTS, WILL HELP US UNDERSTAND
CELLULAR CHARACTERISTICS THAT
COULD CLUE US INTO CELL
PLASTICITY OVER TIME AND MAYBE
WHY I HAD REGENERATE BUT WE ALSO
MAY BE ABLE TO STUDY THE
DIFFERENCE IN HOW THESE CELLS
RESPOND IN ACUTE INJURIES THAT
WE DON'T BELIEVE GO ON TO POST
CHRONIC DISEASE VERSUS ACUTE
INJURIES THAT GO ON TO CHRONIC
DEC SO THERE ARE INHERENT
DIFFERENCES IN HOW THE CELLS ARE
RESHAPED.
NTHEY HAVE PROPOSE THE EVIDENCE
BASED SALES FOR [INDISCERNIBLE]
BASAL CELLS AS WELL.
THAT'S IN HUMAN THAT'S HARD TO
ASK QUESTION BECAUSE OF LACK OF
THESE TOOLS LIKE WE CAN DO THESE
TRACINGS BUT THERE'S ALSO PAPERS
FROM THE GROUP USING AGINOID AND
COCULTURE MODELS WHERE WE
DEMONSTRATE THAD INDEED TYPE 2
CELLS CONVERT TO GENERATED THESE
CELLS AND EVENTUALLY THEY BECOME
BASAL CELLS, SO THERE IS THAT
EVIDENCE AT LEAST FROM THE
CULTURES AND THEY CLEVERLY USE
THIS MODEL WHERE THE MIX MATCHED
UNDERLYING FIBROBLASTS LIKE THEY
USE DIFFERENT FIBROBLASTS AND
THEY SEE 1 BUT NOT WITH OTHER
INDICATING WHEN THERE IS EVIDENT
NEARBY THAT IS INDUCING TYPE 2
CELLS CONVERSION TO THESE
BASALOID AND THEN EVENTUALLY
BASAL CELLS.
SO NOW SPEAKING
DR. [INDISCERNIBLE] HAS RAISED
HAND TO ASK'S QUESTION.
SO YOU CAN GO AHEAD.
>> I WAS -- IT WAS PASSED ALONG
BUYER I WAS READY BUT I WANTED
TO SAY THERE ARE AT LEAST 3
PATHWAYS FROM A TYPE 2 CELL IN
THE HUMAN THAT'S NOT TRUE IN THE
MOUSE, ONLY 1 IN THE MOUSE, THEY
WERE ALL CONCERNED ABOUT
TRANSITIONAL CELLS GOING FROM
TYPE 2 TO TYPE 1 IN THE MOUSE
BUT IN THE HUMAN THEY CAN BECOME
THESE RAS 0 CELLS AND THEY
BECOME TYPE 1 CELLS.
SO I GUESS MY QUESTION WAS
WHETHER OR NOT THERE'S SOME SORT
OF COMPETITION AS TO WHICH
PATHWAY FOLLOWS AND WHETHER THE
DOMINANCE OF THAT PATHWAY JUST
REFLECTS THE PATHWAY THAT'S THE
MOST RESILIENT FOR THAT AREA OF
THE LUNG THE CELLS THAT BEHAVE
THE BEST IN THOSE CONDITIONS AND
THAT WILL BE A DIFFERENT WAY OF
THINKING ABOUT RESILIENCE BUT I
THINK IT'S PLAUSIBLE ANYWAY,
THAT'S WHAT I HAD.
>> GREAT POINT.
I AGREE WITH YOU.
I MEAN THIS ACTUALLY CONNECTS
THE TO LYNN'S QUESTION AND
DAVID'S QUESTION AS WELL, RIGHT?
WHEN AND WHERE THESE CELLS
ACTUALLY TAKE DIFFERENT PARTS
NOMINAL VERSUS PATHOLOGICAL
SMRKS AND WE MADE A POINT TO
PREVIOUS DISCUSSION, WE HAVE
ALSO SHOWN THE LUNG SLICES THAT
AT2s CAN GIVE RISE TO BASAL
CELLS.
SO YES, THEY USE THE FIBROTIC
COCKTAILS INCLUDING LPS AND
OTHER THINGS AND THEY FOUND TYPE
2, YOU KNOW IT'S LIKELY THAT
YOU'RE LOOKING AT THE SLICES AND
YOU FOUND THE BASAL LIKE
SIGNALLURES IN THE CELL.
THAT'S IMPORTANT POINT.
WOULD THIS BE DRIVEN BY CLONE
SELECTION?
YEAH, THAT'S RIGHT.
THEY CAN BE A NONABERRANT STATE,
THAT'S FINE, YEAH, WE NEED MODEL
ISES TO TEST IS ESPECIALLY IN
HUMAN.
>> OH SORRY, I WAS ON MUTE,
SORRY FOR THAT EMPLOY.
>> OKAY, SO THINKING ABOUT YOUR
POINT OF A DIFFERENT PATHWAY,
WHICH PATHWAY THE CELL CHOOSE TO
USE, I WAS THINKING LIKE, A
MULTIPLE PATHWAY HAS BEEN SHOWN
FOR THIS TRANSITION PUSH, BUT IN
MY POINT, MAYBE BIAS LIKE
DIFFERENT GENETIC VARIANCE AND
DIFFERENT PREFERENCE, LIKE THE
WNT AND THE YAPP, AND MANY
DIFFERENT PATHWAYS BUT I THINK
OF INDIVIDUAL PEOPLE, THEY MAY
HAVE DIFFERENT PREFERENCE AND
WHAT WE'RE DOING, IN THE CELL
MODEL AND THE MOUSE MODEL, THERE
ARE ADDITIONAL VERY IMPORTANT.
>> TO ANSWER YOUR QUESTION, I
THINK SOMEONE ASKED A QUESTION
ABOUT THE CLONE IN THESE AND I
THINK IT'S A VERY, VERY
THOUGHTFUL AND IMPORTANT
QUESTION.
AND I THINK THE PLACE THAT
PEOPLE REALLY TALKED ABOUT IT IS
IN CANCER WHERE IT'S OBVIOUS
IMPORTANCE BUT THE REASON I
THINK IT'S SUCH A PRESSIENT
QUESTION IS 1 THE QUESTIONS IN
DISEASE IN GENERAL BUT IN LUNG
DISEASE SPECIFICALLY IS WHY IS
DISEASE PATCHY?
AND 1 THING I WILL SAY FOR
EXAMPLE, THERE ARE PARTS OF
CYSTIC FIBROSIS PATIENT'S LUNGS
LOOK PERFECT AND THE OLD
EXPLANATIONS WE GAVE WERE QUITE
UNSATISFACTORY AND I BEING SAY
THE SAME THING ABOUT IPF AND WE
HAVE SOME EXPLANATIONS NOW,
RIGHT, LIKE PERIPHERY, MECHANO
TRANSDUCTION BUT NOT THE ENTIRE
PERIPHERY LOOKS THE SAME AND
IT'S HARD TO IMAGINE IT'S JUST
LIKE ABSOLUTELY UNIFORMLY JUST,
YOU KNOW, PLURAL PRESSURE,
PULLING ON THE DISTAL LUNG.
SO I THINK THERE'S MORE THERE.
AND I WOULD INTERESTED IN ASKING
ALL OF YOU, I REMEMBER A LONG
TIME AGO, AND I MY MEMORY IS
FADING BUT THERE WAS SOME
QUESTION ABOUT CLONALITY IN IPF
BASED ON SOME KIND OF
CHROMOSOMAL ABERRATIONS.
I THINK IF TIM BLACKWELL IS
HERE, HE WOULD KNOW ABOUT IT,
BUT THERE WAS A LITERATURE TO
SUGGEST THAT THERE WAS CLONALITY
BASED ON CHROMOSOMAL ABERRATIONS
IN TELEORDER OF MICRONS RACE
DEFICIENT DISEASES I THINK AND
IF YOU DON'T KNOW THAT, YOU
MIGHT JUST GO BACK AND LOOK AT
THAT. THERE WAS A LOT OF
SUGGESTION ABOUT THAT.
BUT NOW THERE'S SO MUCH ABOUT
SOMATIC MUTATION AND THERE'S A
GOOD REASON TO THINK TO THINK
THAT SOMATIC MUTATIONS COULD
CAUSE HETEROGENERATED AITY WHICH
MAY CONTRIBUTE SELECTIVE GROWTH
ADVANTAGE BUT NOW LET'S THINK
ABOUT LIKE SELECTIVE IMMUNE
SUSCEPTIBILITY.
LIKE IMAGINE THAT.
AND SO I THINK THAT'S A VERY
INTERESTING QUESTION.
>> YEAH, I MEAN IN THAT LINE, AT
LEAST FROM THE IMMUNE SYSTEMS
SIDE, THERE ARE A COUPLE OF
PAPERS NOW, THE CLONAL HEMEAT O
POETICESIS AND TRANSCRIPTION AND
OF COURSE, I THINK THAT MIGHT BE
A VERY [INDISCERNIBLE]
>> YEAH, EVEN THE FIBROBLASTS OR
PATS CAN HAVE SOMATIC MUTATION
THAT RESISTS OR AMPLIFIES ANY
KIND OF CYTOKINE SIGNAL.
>> YEAH, THAT'S REALLY IMPORTANT
BECAUSE THESE STRANSITION STATE,
THERE IS ABLITERATION IN CANCER
FIELD AS WELL AS IN HUMAN,
SIMILARITATESENTIOUS MERGED BUT
LAKELY DEFINITE FLAVOR BUT THEY
HAVE OVER LAPPING MOLECULAR
FEATURES SO WHENY WOO SEE
NONMALIGNANT DISEASE VERSUS ONO
GENIC EVEN HAPPENS, I THINK THAT
WILL BE GOOD, TOO.
>> YEAH.
>> GOING BACK TO CLONALITY, HOW
DO WE ADDRESS IN BLOOD, LIKE THE
GROUP HAS DONE REALLY NICE WORK
USING THE SINGLE CELL ATACSEQ,
WE HAD TO INVENT TOOLS TO ANSWER
SIMILAR QUESTIONS NBUT YOU WOULD
THINK DISEASE, RIGHT?
GENERAL PROPERTY OF THE DISEASE
MIGHT BE SELECTION OF PATHOLOGIC
CLONES THAT GROW TOO BIG.
RIGHT?
ANALOGY AGAIN TO THE CANCER
FIELD AND THAT IS WHEN YOU HAVE
SELECTIVE, LIKE ADVANTAGE FOR AN
A PERRENT CLONE THAT IS WHEN
YOU HAVE A DISEASE.
IT'S AN THE PATCHY WORST PART OF
THE DEC WHERE YOU SEE THE MOST
MANIFEST EXAMPLE OF IT.
>> THE EXPANSION IS MORE LIKE
RELEVANT TO IPF COMPARED TO
COPD?
>> I THINK IT WILL APPLY
EQUALLY.
I THINK IT'S SUCH A GENERAL
CONCEPT THAT IT COULD APPLY TO
ULTIMATELIA OF DISEASE, REALLY.
>> IF I COULD ENTER INTO IT
DISCUSSION A LITTLE BIT ABOUT
PATCHY DISEASE, YOU KNOW, WE
HAVEN'T TALKED ABOUT THE
ENVIRONMENT AND DISTRIBUTION AND
I CAN'T HELP BUT THINK THAT SOME
OF THE PATCHINESS, CERTAINLY NOT
ALL OF THIS, I HAVE BEEN VERY
INTERESTED IN THIS ISSUE OF
SOMATIC MUTATIONS BUT SOME OF
THE PATCHINESS MAY BE DUE TO
WHAT WE'RE INHALING AND HOW
THOSE MICROSCOPIC RESPONSES ARE
ARE PLAYING OUT IN SPECIFIC
REGIONS OF THE LUNG,
SIMULTANEOUSLY OR CO INSURE
MARKET DENTALLY ACROSS DIFFERENT
AREAS OF HETEROGENEOUS LUNG.
SO IT'S IMPORTANT TO THINK ABOUT
THE ENVIRONMENTAL COMPONENT OF
THIS AS WELL.
>> I THINK DAVID, I AGREE WITH
YOU COMPLETELY AND FURTHER MOW I
WOULD AMPLIFY YOUR COMMENT BY
SAYING THERE'S ACTUALLY VERY
GOOD DATA TO SUGGEST YOU'RE A
HUNDRED% RIGHT, INCLUDING THINGS
LIKE PARTICLE DEPOSITION WHERE
YOU CAN ACTUALLY PRECISELY
QUANTIFY EXACTLY WHAT YOU'RE
SAYING.
AND THEN THERE MUST BE A WHOLE
BUNCH OF OTHER THINGS LIKE WHEN
YOU HAVE AN UNKNOWN ANTIGEN IN
SARCODOSEIS, BUT ALMOST
CERTAINLY ANTIGEN DRIVEN THAT IT
WOULD DEPOSIT IT IN DIFFERENT
PARTS OF THE LUNG.
SO I THINK YOUR STATEMENT DAVID
HAS MORE EVIDENCE THAN ANYTHING
I SUGGESTED WHICH IS JUST PURELY
THEY--THE RETINAL
LOCATIONICICAL.
BUT WE HAVEN'T LOOKED SO WHY
WOULDN'T TB THE CASE TO SOMATIC
MOSAICISM ISN'T THE CASE?
>> YEAH.
>> READING FROM THE COMMENTS 1
FROM SARAH LYNN, HOPE WE WILL
DISCUSSION ENGINEER TRACE MARKER
IN HUMAN CELLS IN THE TECHNOLOGY
SESSION LATER, SO, I THINK
THAT'S A GOOD SESSION TO DISCUSS
ABOUT SOME OF THIS.
HOW DO WE PROBE WHO'S HAPPENING
IN THE HUMAN LUNG.
I MEAN WE HAVE SOME OF THE
SPIRITUAL STRUGGLES ORDER OF
MICRONSETRY AND SOME OF THE LYNN
CLINICAL MEASUREMENTS BUT MORE
FROM A CELLULAR AND TISSUE
LEVEL, HOW DO WE GET TO THAT?
I THINK THAT'S A GOOD SESSION TO
TALK ABOUT EVEN ENVIRONMENTAL
EXPOSURE, YOU KNOW CAN BE
MEASURED HOW MUCH 1 WAS EXPOSED
IN THE LAST 10 YEARS, LET'S SAY
OR THROUGHOUT THEIR LIFE, CAN WE
GET SOME READ OUTS FROM THE
BIOMARKER POINT OF VIEW.
>> BACK TO SOMATIC MUTATION, I
WANT TO DROP DOWN A MEASURE HERE
BECAUSE SHE'S DONE A LOT OF
PHENOMENAL WORK RELATED TO
SOMATIC MUTATION IN
[INDISCERNIBLE] IS IPF.
YEAH, I THINK, JOHN I DON'T KNOW
IF YOU WANT TO UNMUTE YOURSELF
BUT SHE POST HER COMMENT THAT
THE SOMATIC MUTATION IN IPF IS
CLEARLY EVIDENT.
JOHN'S HERE,.
>> YEAH, THANK YOU SO, RIGHT NOW
BECAUSE OF THE LIMITED LUNG
TISSUE WE'VE USED RNA SEQ AND
MATCHED BLOOD WHOLE GENOME
SEQUENCING TO CULTURE MUTATIONS
AND WE SEE ACTUALLY EVIDENTLY
IPT GREATER CREASING SOMATIC
MUTATIONS IN IERK PF LUNGS AND
PRELIMINARY ANALYSIS OF SINGLE
CELL BASED STUDIES SO THAT THE
MUTATION [INDISCERNIBLE] SO
CTHC1 CELLS, THOSE ARE THE CELL
TYPES THAT CARRY MOST OF THE
MUTATIONS.
ACTUALLY THAT'S INTERESTING HAVE
YOU DONE ANY REGIONAL SAMPLES TO
SEE IF THERE ARE RECURRENT OTHER
SIMILAR PATTERNS THAT EMERGED?
>> YEAH, SO, I'M COLLABORATING
WITH DR. SCHWARTZ, SO WE HOPE TO
SAMPLE DIFFERENT AREAS WITH A
DIFFERENT DEGREE OF FIBROSIS,
BUT THE 1S WE CAN CALL SOMATIC
MUTATIONS NOW ARE CLONEALLY
EXPANDED 1S BECAUSE WE'RE
CALLING FROM RNA SEQ, AND WE CAN
ONLY CLONE LARGE MUTATIONS AND
THOSE GENES WE SEE THAT ARE
ENRICHED FOR MUTATIONS ARE
NORMAL GENES IMPORTANT FOR IPF,
MUK5, 4, B, 16 OR EMT RELATED
GENES.
>> YEAH, I MEAN, I ASK THE
REGIONAL SAMPLING BECAUSE IF YOU
TAKE IN UNAFFECTED REGION AND
LET'S SAY FIBROTIC REGION, YOU
KNOW IF THERE ARE PATTERNS THAT
OVERLAP PATTERNS THEN IT CAN
SUGGEST, IT WON'T BE DEFINITELY
BUT IT CAN SUGGEST CLONALITY IF
THERE ARE DIFFERENCES, YEAH.
CAN WE MAKE A DNLIZATION,
BECAUSE WE'RE CANNED TO MAKE A
NEW INTRODUCTION AND IT MADE ME
REALIZE THAT SOME OF US HAVE A
CLINICAL BACKGROUND AND SOME OF
US HAVE A Ph.D. BACKGROUND AND
WE'RE BOTH TRYING TO CONVERGE ON
THIS BUT NO 1 PERSON CAN DO
EVERYTHING.
AND EVEN WHEN YOU CAN BOTH LOOK
AT THE OTHER TO TRY TO IMPORT
SOMETHING, IT WOULD BE VERY
INTERESTING FOR EXAMPLE, TO FUSE
LIKE A CLINICAL RESEARCHER, WHO
IS ACTUALLY DOING A CLINICAL
TRIAL AND COMBINING IT WITH
SOMEONE MORE ON THE BASIC SIDE
THINKING ABOUT DISEASE AND MAYBE
THAT IS A DIRECTION WE SHOULD GO
IN TO ACTUALLY ENCOURAGE
COLLABORATION ACROSS THE
CLINICAL RESEARCHERS AND THE
BASIC RESEARCHERS.
IT'S NOT EASY TO DO BUT IT'S THE
KIND OF THING, THE NIH CAN DO,
THE VERY OTHER FEW ORGANIZATIONS
CAN.
YEAH, THANKS WE ARE RUNNING LATE
FOR THE SCHEDULE.
THANKS EVERYONE FOR THESE LIKE
VERY INTRIGUING DISCUSSION AND
THANK ALL OF SPEAKERS, IT'S
REALLY GREAT OPENING SESSION FOR
OUR 2 DAY SYMPOSIUM.
OUR BREAK TIME IS OH HALF HOUR,
SO, TEAM WE NEED TO COME BACK AT
HALF AN HOUR LATER, IT'S 1:15
STARTING SESSION 2.
NYES.
WE WILL BE BACK AT 1:15.
THANK YOU.
WE ALSO HAVE LOTS OF DISCUSSION,
WE DIDN'T HAVE TIME BUT WE ARE
COMING BACK, CONTINUING
TOMORROW, AND WE HAVE STRATEGIC
SCHEDULE TOMORROW, IN THE LAST
SECTION.
>> THANKS EVERYONE.
>> THANK YOU EVERYONE.
WE WILL HEAR FROM 4 SPEAKERS
TODAY.
NEXT SLIDE, PLEASE.
WHO WILL TRY TO TALK ABOUT THE
NEW TECHNOLOGIES THEY ARE
DEVELOPING AND HOW THEY CAN BE
USED TO STUDY PROGENITOR AND
TEMCELL BEHAVIOR IN THE LUNG AND
DURING THE DISCUSSION WE WILL
FOCUS ON THE CRITICAL RESEARCH
GAPS, AND WHAT WE DO TO ADDRESS
THAT.
NEXT PLEASE.
I THINK THESE SLIDES WILL BE FOR
DISCUSSION.
WE WILL GO TO SPEAKER #1 EMPLOY
FIRST SPEAKER IS DR. MING
XIA GU, PHYSICIAN SCIENTIST AT
CUSTOM CENTER AT CHILDRENS, SHE
IS IN THE PEDIATRICS DIVISION
THERE, SHE WILL TALK TO US ABOUT
IPSC PROGENITOR CELLS AND HOW WE
CAN USE ORGANOID MODELS?
THANK YOU FOR THE KIND
INTRODUCTION AND THANK YOU FOR
HAVING ME TODAY.
I REALLY, REALLY ENJOYED THE
MORNING SESSION AND ESPECIALLY
THE DISCUSSION AND THE QUESTIONS
RAISED BY THE PANEL IS VERY
INSIGHTFUL AND THOUGHTFUL AND
FOR US, WE ARE INTERESTED IN
DEVELOPING HUMAN INDUCED
PLURIPOTENT STEM CELLS TO
ORGANOID MODELS TO HOPEFULLY
ANSWER SOME OF THE REMAINING
QUESTIONS RAISED BY THE PANEL
THIS MORNING AND ALSO TO PROVIDE
A HUMAN PERSPECTIVE IN DISEASE
DEVELOPMENT.
AND POSSIBLY UNDERSTANDING, SOME
FOR EXAMPLE, THE PATCHY BIOLOGY
MENTIONED EARLIER AND
POTENTIALLY CRISR THE SYSTEM AND
INTRODUCE SOMATIC MUTATIONS AND
UNDERSTAND CLONAL EXPANSION IN
INVITRO MODELING SYSTEM.
ALL RIGHT, NEXT.
SO TODAY, DUE TO THE TIME LIMIT,
I WILL JUST INTERVIEWS 1 OF OUR
RECEIPT STUDIES FOCUSING ON
ALVEOLAR CAPILLARY DYSPLASIA
WITH MISALIGNMENT OF PULMONARY
VAINS WHICH IS A CONGENITAL LUNG
DISORDER WITH NO CURE AND AS YOU
CAN SEE FROM THE STAINING, THE
DECREASED NUMBER OF PULMONARY
CAPILLARIES AND ADJACENT TO
SMALL PULMONARY ARTERIES AND
LOBULAR UNDERDEVELOPMENT THIS IS
CAUSED A MUTATION IN THE FOX F1
AND EXPRESSED IN ENDOTHELIAL AND
MESSENTERIEN CHIME AT
POPULATIONS AND IT LED TO STUDY
ACD/MPV IN A CULTURE DISH.
NEXT.
THIS CONDITION IS CAUSED BY A
MUTATION IN FOX F1 SO THROUGH
THE COLLABBUATION WITH THE LUNG
MAP GROUP HERE THEY HELPED ME
LOOK INTO THE EXPRESSION PATTERN
OF FOX F1.
NEXT.
SO THE MAGENTA COLOR INDICATES
THE CELLS EXPRESSING FOX F1
DURING LUNG DEVELOPMENT AND
CLEARLY THERE'S AN ENRICHMENT IN
THOSE 2 CAPE ILLEGALSARY
ENDOTHELIAL CELLS, CAP 1 AND 2.
AND I ALSO WANTED TO DRAW
ATTENTION TO THE [INDISCERNIBLE]
POPULATION, AT1 AND 2 AS THERE'S
NO EXPRESSION OF FOX F1 IN THOSE
EPITHELIAL CELLS.
NEXT.
SO THE INTERESTING PHENOTYPE
USED IN THESE SINGLE NUCLEI
RNA-ATAC-SEQ CASES IS THAT
MINZHE AND JEFF, NOT ONLY FOUND
A REDUCTION IN THE CAPILLARY
CELLS, THEY ALSO OBSERVED
ABNORMALITIES IN EPITHELIAL
DIFFERENTIATION AS SHOWNOT RIGHT
HAND SIDE, THEY FOUND AND THIS
IS ALSO A PHENOTYPE SEEN IN A
VARIETY OF LONG INJURY MODELS.
THERE'S ACCUMULATION OF THIS
INTERMEDIATE CELL TYPE OF BOTH
AT1 AND 2, SIGNATURES IN THE
CASES AND A REDUCTION IN AT1
CELL POPULATION IN THE PATIENT
WHICH INDICATES THAT THESE
ABNORMALITIES IN THE EPITHELIAL
CELLS ARE LIKELY CAUSED BY THE
ABNORMAL CAPILLARY DEVELOPMENT
AS THE EPITHELIAL CELLS DO NOT
EXPRESS FOX F1, THEREFORE WE AIM
TO DEVELOP A NEXT GENERATION OF
VASCULARIZED LUNG ORGANOID
MODELS TO STUDY THE ALVEOLI
CAPILLARIES AND CROSS TOCK IN
ACD/MPV.
NEXT.
SO DEVELOPING SUCH COMPLEX
ORGANNAL MODEL IS DIFFICULT
BECAUSE DURING AT1 AND 2
EPITHELIAL CELLS ARE DERIVED
FROM THE EBD O THERMAL LIPIAGE
WHEREAS THE VASCULAR ARE DERIVE
FRIDAY THE MESODERM AND IN THE
DEFERENTIATION, TO REQUIRES HIGH
MBP, WITH ACTIVATION WERE
GENERATING [INDISCERNIBLE]
ENDODERM REQUIRES LOW BMP BUT
HIGH TGF BETA AND FGF.
THAT MADE US TO FIRST TRY TO
BALANCE ALL THESE SIGNALING
PATHWAYS SO WE CAN MAINTAIN BOTH
MESODERM AND ENDODERM
POPULATIONS IN THE SAME SPHEROID
TO EVENTUALLY VASCULARIZE THE
ORGANOIDS.
NEXT, SLIDE, PLEASE.
SO, THE NEXT COUPLE OF SLIDES,
I'M GOING TO BRIEFLY SUMMARIZE
THIS WORK SPEARHEADED BY SIMON
IN THE LAB TOGETHER WITH
[INDISCERNIBLE] VICTOR, SO WHAT
THEY FOUND IN THE PAST 4 YEARS
OR SO, THEY'VE BEEN TRYING TO
REALLY HARD TO MAINTAIN MULTIPLE
LINEAGES IN A SINGLE SPHEROIDS
TO MAINTAIN DURING DEVELOPMENTAL
PROCESS.
SO OUR FIRST POPULATION IS WHEN
SIMON FOR THE INCREASE THE BMP
ACTIVATION DURING THE FIRST 3
DAYS OF SPHOAROID
DIFFERENTIATION, HE FOUND HE
COULD INCREASE THE POPULATION OF
THE MESODERMAL CELLS AND SHOW
ONCE THEY MEET HERE AT THE
BOTTOM, AND AT THE SAME TIME
MAINTAIN THE ENDOTERM POPULATION
STAYING POS 5 FOR FOX F2 IN THE
GREEN, SO USING THIS FINE TUNING
METHOD, WE MAINTAINED THE BOTH
ENDODERM AND MESODERM LINEAGES
THROUGH THE SAME SPHEROIDS
THROUGH DIFFERENTIATION AND FOR
THE NEXT 4 DAYS OUR SORE, WE
PUSHED THE ENDODERM
MALPOPPULATION FORWARD THE
EPITHEL YIEWM, AT THE SAM TIME
MAINTAINING METRICS SENK MALAND
ENDOTHELIAL CELLS, IF YOU CLICK
ON THE IMAGE ON THE RIGHT SIDE,
IT'S A 3D MODEL.
IN THE MITTED YOU'RE SEEING THE
FOREGUT, AND THEN ALSO THE
MESODERM POPULATION AND ALSO
CD31 POSITIVE BLOOD VESSELS IN
WHITE.
ALL RIGHT, NEXT SLIDES.
AND THEN, WE FURTHER
DIFFERENTIATED THE FOREGUT
LINEAGE TOWARD THE LUNG
PROGENITOR ORGANOID WITH
MESSENTERIEN CHIME AND
VASCULATURE.
SO THIS A BRIGHT VIEW LINEAGE OF
ORGANOIDS AND YOU CAN APPRECIATE
THAT MAJORITY OF THE CELLS ARE
ENDOTHELIAL CELLS.
COMPARED TO -- THEY HAVE THE
POSITIVE EPITHELIAL CELLS IN THE
MIDDLE, FOX P1 POS55 TELES IN
THE CD31 POSITIVE VESSELS AND ON
THE RIGHT HAND SIDE THESE ARE
VASCULARIZED ORGANOIDS AFTER 21
DAYS OF DIFFERENTIATION FROM
HUMAN IPSCs, AND THEY
SELF-ARAINCHL INTO THIS
STRUCTURE WITH THE EPITHELIAL
CELLS IN THE MIDDLE AND
MESENCHYMAL CELLS IN THE
EMTHELIUM AND THAT'S ALSO
WRAPPED BY THE CD31 POSITIVE
VESSELS.
NEXT SLIDE, PLEASE.
AND TOO, FOR THE MODEL OF THE
ALVEOLAR STRUCTURE WE
DISTALLIZED THE ORGANOIDS USING
THE PARTICLE DEVELOPED BY
[INDISCERNIBLE] GROUP AND WE
FOUND THAT THE CELLS AT A
POSITIVE FOR DISTAL MARKER FOX 9
AND ALSO AT1 AND AT2 MARKER
GENES IN THESE ORGANOIDS AFTER
DISTALLIZATION.
NEXT SLIDES.
AND THIS IS A 3D RECONSTRUCTION,
[INDISCERNIBLE] THIS IS TO GIVE
YOU KNOW OVERVIEW OF THE
INTERACTION OF THE ENDOTHELIUM
AND EPITHELIAL CELLS.
WE ARE ABLE TO,A CHIEF IN THESE
ORGANOID MODELS SO IT'S SOMEWHAT
SIMILAR TO THE LUNG STRUCTURE
WITH THE EPITHELIAL CELLS IN
ORANGE, AND WE INTRODUCED A
DESCENT AMOUNT OF THE CD31
POSITIVE BLOOD VESSELS AND I
ALSO WANT TO IMAGINE THESE ARE
FEASIBLE AND THEY HAVE LUMEN
THAT'S RECAPITULATING A
CAPILLARY SIZE.
NEXT SLIDE, TO CHARKTIZE THE
ORGANOIDS WE PERFORM SINGLE
CELL RNASEQ TO THE ORGANOIDS BUT
AS YOU CAN SEE FROM THE DOT
PLOTS WE HAVE A VARIETY OF CELL
TYPES IN THESE ORGANOIDS.
IN ADDITION TO THE EPITHELIAL
CELLS IN OBSERVED IN THE
CONVENTIONAL ORGANOIDS, WE NOW
HAVE MESENCHYMAL IN THE
ENDOTHELIAL CELLS AND I ALSO
WANT TO MENTION THAT AS YOU CAN
SEE FROM THE ORANGE GENES,
HPGD IN KITS THESE ARE LONG
SPECIFIC ENDOTHELIAL CELL
MACROIGEP GENES EXPRESSED IN
THESE ORGANOIDS AND FOX F1 ARE
ALSO ENRICHED IN THESE
MESENCHYMAL CELLS WHICH
INDICATES WHEN WE CO DEVELOPED
MESSENTERY ILLEGALS DERM WITH
THE TISSUE RELEVANT ENDODERMAL
POPULATION THAT MESSY DERMAL
CELLS INCLUDING THE VASC O
ENDOTHELIUM AND THE MESENCHYMAL
CELLS GAINED ORGAN SPECIFICITY
WHICH WILL HELP US BETTER
ADDRESS THE DISEASE IN THE HUMAN
DEVELOPMENT IN THE TISSUES
SPECIFIC MANNER.
NEXT SLIDE PLEASE.
WHAT'S MORE INTERESTING TO US,
IS THAT WE FOUND, WE'RE NOT ONLY
GENERATING DIFFERENT CELL TYPES
IN THESE ORGANOIDS, THE CELL
PROPORTION IS ALSO VERY SIMILAR
TO WHAT'S BEING SHOWN IN HUMAN
FETAL LUNG BASE ON THE RNA SEQ
THAT'S PUBLICLY AVAILABLE, SO
COMPARED WITH THE CONVENTIONAL
LUNG ORGANOID SHOWING ON THE
LEFT-HAND SIDE, THEY HAVE
EPITHELIAL CELLS UNCONTROLLED
DIFFERENTIATION, AND
CONTAMINATION IN THE SYSTEM, SO
IN THE MITSD THEE ARE THE
VASCULARIZED LUNG ORGANOIDS THAT
WE GENERATED AND FIRST OF ALL WE
HAVE A LARGER MOUND OF
MESENCHYMAL POPULATION, DUE TO
THE MAINTENANCE OF THE
MESODERMAL POPULATION IN THE
SPHEROIDS AND WE ALSO HAVE THE
ENDOTHELIAL CELLS AS YOU CAN SEE
IN YELLOW HERE IN THESE
ORGANOIDS AND PROPORTIONALLY
THEY ARE QUITE SIMILAR TO HUMAN
FETAL LUNG.
WE ALSO HAVE MIN NUM
CONTAMINATION OF THE INTESTINE
EPITHELIAL CELLS IN THESE LUNG
ORGANOIDS.
NEXT SLIDE PLEASE.
BECAUSE IN THESE ORGANOIDS ARE
STILL IN SPHEROIDS, WHICH IS
DIFFERENT FROM THE ALVEOLAR
STRUCTURE, SO WE'RE ALSO TRYING
TO BETTER YOU KNOW DRIVE THE
CELL FATE COMMITMENTS AS WELL AS
RECAPITULATING THE ALVEOLAR
STRUCTURES USING ENGINEERING
APPROACHES, SO THROUGH
COLLABORATION, [INDISCERNIBLE]
AT WOMEN'S WE ASSOCIATED THESE
ORGANOIDS AND THEN RECEDE THEM
ON A LUNG OR CHIP MODEL MADE BY
[INDISCERNIBLE].
SO NEXT SLIDE.
WE STAINED BOTH ENDOTHELIAL AND
EPITHELIAL CELLS IN THESE
SCAFFOLDS, AND YOU CAN
APPRECIATE THAT THEY NOW FORM --
ALTHOUGH THEY WERE BEING
DISASSOCIATED INTO SINGLE CELL
SUSPENSION, AND THEN RECEIVED ON
THE LONG SCAFFOLD, THEY ACTUALLY
AGAIN, SELF-PATTERN INTO THIS
STRUCTURE WHICH IS ALSO SIMILAR
TO A MOUSE LONG STRUCTURE, SO IN
THE MIDDLE, THESE ARE ALL
EPITHELIAL CELLS AND THAT'S
WRAPPED UP BY THE CD10 POSITIVE
BLOOD VESSELS AND THIS WORK IS
STILL PRELIMINARY AND WE'RE NOW
ALSO TRYING TO FURTHER OPTIMIZE
THE SYSTEM AT THIS POINT.
NEXT SLIDE, PLEASE.
NOW WE HAVE A VASCULARIZED
KD--SALLY WHO MODEL, WE SUES
THAT TO SEE IF WE COULD
RECAPITULATE SOME OF THE
PHENOTYPES IN PATIENT LUNG
TISSUES USING THESE ORGANOID
MODELS.
THROUGH COLLABORATION WITH
DR. KOTTON AND DR. ROTTIER, WE
OBLIGATIONS STAINED IPSCS FROM
THESE PATIENTS AND THEY CARRY
DIFFERENT TYPES OF MUTATIONS.
ONE IS A BIG CHUNK
[INDISCERNIBLE] IN THE MUTATIONS
AND BINDING DOMAIN OF THE FOX
HEAD 1 PROTEIN.
NEXT SLIDE.
SO FIRST, WE EXAMINED THE
ENDOTHELIAL POPULATION IN THESE
ORGANOIDS AND WE FOUND IN THOSE
ACD/MPV, ORGANOIDS THERE'S
REDUCTION IN THE CD31 POSITIVE
CELLS AND ALSO REDUCED MEMBERS
OF CAP 1 AND CAP 2 ENDOTHELIAL
CELLS IN THE SYSTEM.
NEXT SLIDE.
MORE IMPORTANTLY BECAUSE WE HAVE
MULTIPLE CELL TYPES IN THESE
ORGANOIDS THAT ARE
RECAPITULATING THE CELL-CELL
INTERACTION WE OBSERVED
ABNORMALITY IN THE EPITHELIAL
POPULATION ALTHOUGH THEY DON'T
EXPRESS FOX F1, SO IN THE
ACD/MPV ORGANOIDS THERE'S
ACCUMULATION OF THE INTERMEDIATE
CELL TYPE AT1/AT2.
NEXT.
-- OBSERVED IN THE CONVENTIONAL
ORGANOIDS.
NEXT, PLEASE.
SO IN SUMMARY, WE DEVELOPED
MESODERM ENDODERM MULTILINEAGE
LUNG ORGANOIDS AND WE ALSO
ADOPTED LUNG SPECIFIC GENE
SIGNATURES AND THESE ORGANOIDS
CAN BE FURTHER PATTERNED ON A
IMRKS ELMA SCAFFOLD TO
CAPITULATE THE HUMAN ALVERMEN
INFECTED O LIAISON SYSTEM AND
ALSO THESE ORGANOID PROVIDE
HUMAN-SPECIFIC MODEL TO STUDY
THE DYNAMIC CELL-CELL
INTERACTIONS AND CELL FATE
TRANSITION DURING LUNG
DEVELOPMENT AND DISEASE SUCH AS
ACDMPV, NEXT, IF I STILL HAVE A
MINUTE OR SO, I THINK THIS IS
SORT OF TRYING TO SET THE STAGE
FOR THE DISCUSSION LATER, AFTER
THE SESSION, WHAT ARE THE NEXT
STEPS, RIGHT?
I THINK SARAH ALSO MENTIONED IN
THE MORNING WE CAN APPLY SOME
ENGINEERING APPROACHES OR
BIOLOGY APPROACHES TO TRACE THE
CELLS POSSIBLY INVITRO AND TRY
TO ANSWER SOME QUESTIONS THAT
ARE DIFFICULT TO BE ANSWERED IN
MOUSE MODELS.
SO FOR US, THE NEXT GOAL OR
FUTURE DIRECTIONS IS TO BETTER
-- TO GENERATE A BETTER LUNG
ORGANOID MODEL THAT CLOSELY
RESEMBLES THE HUMAN LUNG
STRUCTURE AND DEVELOPMENTAL
PARALYSIS.
SO THERE ARE SEVERAL PIECES IN
MY OPINIONS THAT ARE STILL
MISSING IN THE CURRENT SYSTEMS
AND I'M ALSO HAPPY TO TALK ABOUT
THAT LATER INCLUDING, YOU KNOW
USING LUNG SPECIFIC ECM TO DRIVE
SOME OF THE LUNG PLATE
DETERMINATION AND USE REPORTER
OR TRACER TO VISUALIZE THE CELL
DIFFERENTIATIONS, THAT'S NOT
ENTIRELY A BLACK BOX TO US AND
IT'S BETTER CONTROLLED AND A
POTENTIAL REPRODUCE ABILITY OF
THE SYSTEM AND ALSO INCORPORATE
A PROPER GRADIENT OF THE MORPHIA
GENERATED TO INDUCE CELL
PATTERNS PROLIFERATION AND
LASTLY, RESULTS OF SOMETHING
WE'RE TRYING TO OPTIMIZE IN THE
CURRENT SYSTEM IS TO INTRODUCE
BIOPHYSICAL CUES SUCH AS FLOW,
IN THE VASCULATURE OR MECHANICAL
FORCE THROUGH THE STRETCHING OF
THE MEMBRANE TO UNDERSTAND HOW
THOSE FACTORS PLAY A ROLE IN
LUNG DEVELOPMENT OR IN RESPONSE
TO LUNG INJURIES.
I THINK THAT'S IT.
NEXT.
I THINK ACKNOWLEDGMENTS SLIDE.
THESE ARE THE PEOPLE CONTRIBUTED
TO MAIORITY OF THE WORK THAT I
TALKED ABOUT TODAY.
SIMON, VICTOR, AND NICOLE.
I ALSO WANT TO THANK TALL MY
COLLABORATORS AT CHILDREN'S AND
OTHER UNIVERSITIES.
HAPPY TO TAKE QUESTIONS LATER
ON.
THANK YOU VERY MUCH.
>> THANK YOU DR. GU FOR THAT
PHENOMENAL TALK NEXT WE HAVE
DR. BIN ZHOU, I THINK THE
DESCRIPTION HERE IS NOT CORRECT,
HE IS A DISTINGUISHED PROFESSOR
AT THE CHINESE ACADEMY OF
SCIENCES HERE TO TALK TO US
ABOUT USING MULTIPLE REPORTER
SYSTEMS FOR HIGH FIDELITY
LINEAGE TRACING IN THE LUNG.
>> OKAY, THANK YOU FOR THE
INTRODUCTION.
CAN YOU HEAR ME?
>> YES, WE CAN HEAR YOU
DR. ZHOU.
>> OKAY, TODAY I WILL INTRODUCE
YOU SOME STORY ON THE RECOMBIN
ACES LINEAGE TRACING TO APPLYING
IT TO STARTING OF LUNG
EPITHELIAL CELL IN LUNG
REGENERATION REPAIR.
NEXT.
SO MY LAB INTRODUCED THE CRE-LOX
P AND DRE-ROX SO WE CAN MORE
PRECISELY DO LINEAGE TRACING.
NEXT, IN THE LUNG THERE HAS BEEN
A DEBATE ON THE ROLE OF
BRONCHIAL ALVEOLAR STEM CELL
BASE THAT IS LOCATED IN THE DARK
JUNCTION BETWEEN THE BRONCHIOLES
AND ALVERMEN INFECTED O LIE, SO
WE WANT TO PRECISE TARGET THIS
BASE IN VIVO.
NEXT.
WE FOUND THAT THIS BASCs,
LABELED BY [INDISCERNIBLE] CAN
CONTRIBUTE TO THE BRONCHIAL
EPITHELIAL CELLS OF THE
BRONCHIAL INJURY.
ALL CONTRIBUTED TO ALVEOLAR
EPITHELIAL CELLS AFTER ALVEOLAR
INJURY.
SO THIS MAKES MULTIPOTENT STEM
CELLS IN
CELLS IN VIVO.
NEXT.
FOR THE ORIGINS OF ALVEOLAR OR
AT2 CELLS IN HOMEOSTASIS AND
DISEASES, THERE HAS BEEN MULTILE
SOURCES IN ADDITION TO THE AT2
CELL PROLIFERATION OR CELL
RENEWAL THAT HAS BEEN REPORTED
THAT CLUB CELLS IN THE BRONCHIAL
CAN TRIBUTE TO AT2 CELLS,
BASCs HAVE BEEN REPORTED TO
CONTRIBUTE TO AT2 CELLS OR EVEN
THOSE AT1 CELLS CAN REVERSE BACK
AND DIFFERENTIATE TO ITS
PARENTAL AT2 CELLS.
NEXT.
NEXT, HOWEVER, THERE ARE SOME
ISSUES ON THE LINEAGE TRACING
FOR THIS PREVIOUS STUDIES.
THE MARKERS FOR AT1, LIKE HOPX,
OR AGER, WE FOUND THESE MARKERS
ARE NOT THAT SPECIFIC FOR AT1
CELLS FOR EXAMPLE, HOPX TARGET
NOT ONLY AT1 CELLS BUT ALSO A
PORTION OF CLUB CELLS.
ATM IS BASED AND THE AT2 CELLS
IN ADDITION TO AT1 CELLS, AND
THE CONVENTIONAL TOOLS FOR CLUB
CELLS LIKE ACTB1, TARGET BOTH
CLUB CELLS AND SFTPC WHICH HAS
BEEN WIDELY USED FOR TARGETING
AT2 CELLS ACTUALLY TARGET
BASCs AS WELL, SO ALL THESE
TOOLS ARE NOT EXCLUSIVELY
TARGETED.
ONE CELL TYPE.
NEXT.
SO WE WANT TO APPLY DUAL
RECOMBIN ACES AND CELL TYPES FOR
EXAMPLE, WE FIRST USED DUAL
RECOMBINNATE TO TARGET AT1 AS I
INTRODUCED HOPX AND AGER ARE NOT
SPECIFIC TO AT1.
BUT THE COMBINATION OF THEM, WE
USE INTERSECTIONAL PART OF
AT-HOPX AND AGER FOR THE CELLS
EXPRESSING BOTH MARKERS WE FOUND
THAT THEY ARE QUITE SPECIFIC,
RESTRICTED TO AT1 CELL LINEAGES.
SO, WE USE IRPT SECTIONAL
GENETICS TO TARGET AT1 CELLS BY
HOPX BY EXPRESSING HOPX AND
AGER, USING THOSE DOUBLE LOX,
AND CROSS IT WITH HOPX AND AGER
AND CRE, SO AFTER
[INDISCERNIBLE] INJECTION, THEY
REMOVE IT LEADING TO EXPRESSION
OF TD-TOMATO.
NEXT.
THE IMAGES ON THE UPPER PANEL IS
A STRATEGY USING HOPX CREA, YOU
CAN USE THAT AFTER THE BLE
OMYCIN INJURY, YOU CAN SEE THE
TDTOMATO, CAN YOUEE THAT SOME
EXTENSION NUMBER OF CLUB CELLS
IN THE BRONCHIALS ALSO LABELED
BY HOPX CRE, SO WE CAN CANNOT
TELL YOU THESE TDTOMATO POSITIVE
OR AT2 CELLS ARE DERIVED FROM
AT1 CELLS OR FROM CLUB CELLS.
IN THE IMAGE S ON THE LOWER
PANEL, WE FOUND THAT THIS TD
TOMATO POSITIVE CELLS LABELED BY
THE DUAL RECOMBIN ACES ARE
SPECIFIC AT-1 CELLS AND THEY DO
NOT CONTRIBUTE TO HOP SFTC AT2
CELLS AFTER THE BLEOMYCIN
INJURY.
NEXT.
WE ALSO PERFORMED PNEUMONECT
ME INJURY AND FOUND THAT AT1
CELLS DO NOT CONTRIBUTE TO AT2
CELLS COMPARED WITH HOPX, CREE,
AND CONTROL.
NEXT.
THEN WE USED DURANNAL RECOMBIN
ACE TO SPECIFICALLY LABEL CLUB
CELLS, AT2 CELLS AND THE BASCs
SIMULTANEOUSLY IN 1 MOUSE, IN 1
MOUSE.
WE USE TRIP O LIGHT REPORTER AND
ACROSS WITH SDG1 AC1 CRE AND
STP3, IN THIS MODEL, CLUB CELLS
IS LABELED BY TD TOMATO, AT2
CELLS ARE LABELED BY
[INDISCERNIBLE] IS THE BASE THAT
EXPRESS BOTH SFTPC AND SFTPC,
AND ADG101 ARE LABELED BY
TD TOMATO AND [INDISCERNIBLE]
EMPLOY SO THEY'RE SHOWING THE
YFP, SO WE COULD LABEL THESE 3
DIFFERENT POPULATIONS WITH 3
DIFFERENT COLORS.
NEXT.
NEXT, SO THIS A DETAILED
STRATEGY, WE LABELED.
WE USED TRIPOLITE REPORTER SO
THIS COULD RESPOND TO CRE OR DRE
RESULLING IN TD TOMATO KREEN AND
WE CALL THEM CLUB TRACER, AT2
TRACER AND BASC TRACER AND IN
THE RIGHT, LEFT, AND RIGHT LOWER
PANEL YOU CAN SEE THAT WE CAN
LABEL THIS DISTINCT EPITHELIAL
POPULATION THAT WOULD BE
DIFFERENT COLORS FOR US AND THE
REPORTERS.
NEXT.
THEN WE INDUCED MICE TO INDUCE
MICE INJURY COMPARED WITH THE
KD--SALLY SHAM INJURY, COMPARED
WITH THE SHAM, THE UPPER PANEL
IS THE SHAM, YOU CAN SEE THE
COLORED CELLS ARE TD TOMATO
POSITIVE, THE AT2 CELLS ARE
GREEN, AND THE BASE WITH THE
REGION EXPRESSING BOTH TD TOMATO
AND THE CSGREEN, THEY ARE OF THE
BLEOMYCIN INJURY, WE OBSERVE
THAT SUBSET OF SFTP2 CELLS
EXPRESS YFP OR EXPRESS TD TOMATO
INDICATING THEY ARE DERIVED FROM
THE CLUB CELLS.
ON THE RIGHT PANEL THESE ARE THE
DATA QUANTIFICATION DATA FOR THE
CELL AT2 CELLS THAT DERIVED FROM
CLUB, BASCs OR SELF-RENEW
AFTER BLEOMYCIN.
NEXT.
SO WE FOUND THAT AFTER BLEOMYCIN
INJURY, IN ADDITION TO THE
SELF-RENEWAL OF AT2, AIT 2
CELLS, CLUB CELLS AND BASCs
CAN DIFFERENTIATE INTO AT2 CELLS
AND OUR TRIPLE COLOR LABELED IT
GENETIC LINEAGE TRACING PRECISE
QUANTIFICATION OF THEIR
CONTRIBUTION OF THE INJURY.
NEXT.
WE CAN AWLINGS MANIPULATE THE
SYSTEM AND OVER EXPRESS P21 AND
GFP, SO THAT REGENERATIVE
ABILITY OF BASCs AND AT2 CELLS
ARE IMPAIRED AS THEY CANNOT
PROLIFERATE ANYMORE AFTER INJURY
BECAUSE THEY OVEREXPRESS
PROLIFERATION INHIBITOR MARKER
P21.
NEXT.
IN THIS MODEL, WE FOUND THAT AWE
SUBSTANTIAL NUMBER OF TD TOMATO
POSITIVE CELLS WERE AT2, WERE
SFTPC, POSITIVE, AT2 CELLS, THEY
CONTRIBUTE TO AT2 CELLINGS AFTER
4 WEEKS.
NEXT.
AT 8 MONTHS, WE FOUND THAT AS A
MAJORITY OF AT2 CELLS ARE NOW
TDTOMATO INDICATING THAT UNDER
SPECIAL CONDITIONS WHEN AT2 CELL
REGENERATIVE ABILITY IS
IMPAIRED, CLUB CELLS CAN
CONTRIBUTE TO THE MAIORITY OF
AT2 CELLS CELLS SUGGESTING THEH
PLASTICITY FOR REGENERATION FOR
CLUB CELLS TO AT2 CELLS.
NEXT.
WE CAN ALSO MANIPULATE THE
GENETIC SYSTEM TO STUDY THE
MOLECULAR MECHANISMS OF CLUB
SALES, OR BASCs, THEY ARE
DIFFERENTIATION INTO AT2 CELLS.
WE, BY SINGLE CELL RNASEQUENCE
OF THESE TRACES, WE FOUND THE
DYNAMIC CHANGE OF NOTCH
SIGNALING PATHWAYS, SO WE DELETE
NOTCH TRANSCRIPTION FACTOR RPG
IN BASCs AND CLUB CELLS AND
EVALUATED THESE 2 POPULATIONS
DIFFERENTIATION ABILITY TO AT2
CELLS, AND THE UP PANEL, WE USE
THIS USE THIS FLOX AS ELIMINATED
CONTROL AND WE USE RBGFLOX AS A
KNOCK OUT AND WE FOUND TO
COMPARED TO THE CONTROL IN THE
MUTANT THERE ARE INCREASED WILD
[INDISCERNIBLE] CLONES AND
REDUCED TD TOMATO POSITIVE
CLONES FOR AT-2 CELL
CONTRIBUTION ON THE RIGHT PANEL,
WE CAN SEE THE QUANTIFICATION
DATA OF THIS IMMUNOSTAINING DATA
AND ALSO THE FACTS DATA SHOWING
THAT AFTER NOTCH KNOCK OUT, THE
CONTRIBUTION OF CLUB SALES TO
AT2 SALES AND SIGNIFICANTLY
REDUCE THE AND 1 OF THE
CONTRIBUTIONS TO AT2 CELLS ARE
INCREASED SUGGESTING THAT NOTCH
SIGNALING DISTINCTLY REGULATED
THESE 2 PROGENITOR CELLS,
DEFERENTIATION TO AT2 CELLS.
NEXT.
IN SUMMARY, OUR STUDY SHOWED
THAT THE AT1 CELLS ARE QUITE
STABLE DIFIENTIATIONS, THEY DO
NOT REVERSE BACK TO THEIR
PARENTAL CELLS UNDERSPECIFIC
INJURY, DURING HOMEIO STASES WE
FOUND THAT IN ADDITION TO THE
AT2 CELLS SELF-RENEW CLUB CELLS
AND BASCs CAN CONTRIBUTE TO
AT2 CELL SAYS AND THEY ARE
DIFFERENTIATION TO THE AT2 CELLS
THAT ARE DISTINCTLY REGULATED BY
NOTCH SIGNALING.
NEXT.
IT HAS BEEN REPORTED THAT UNDER
SPECIAL CONDITIONS AT1 CELLS CAN
DIFFERENTIATE INTO AT2 CELLS AND
EVEN AT1 CELLS CAN CONTRIBUTE TO
THE LUNG ADEN O CARCINOMAS IF
THEY EXPRESS KRAS, SO WE IN
FUTURE RE, VALENTINEDUATEED
THESE STUDIES BY USING DUAL
RECOMBIN ACES MEDIATED LINEAGE
TRACING TO UNDERSTAND THE
PLASTICITY OF WHAT AT1 CELLS AND
THE SPECIAL CIRCUMSTANCES.
NEXT.
AND FINALLY I WOULD LIKE TO
THANK MY STUDENTS,
COLLABORATORS, AND FUNDERS.
THANK YOU.
THANK YOU DR. ZHOU.
I HAD THE PLEASURE TO INTRODUCE
DR. RUOBING WANG WHO IS A AN
ASIFT ANT PULL MONITORROLOGYIST
ASSISTANT AT HARVARD MEDICAL
SCHOOL.
DR. WANG DOES FANTASTIC WORK IN
IPSCs CELLS AND SHE WILL TALK
TO US ABOUT LEVERAGES IPS AIR
WAY TO INVESTIGATE PULMONARY
IONOSIGHTS AND CYSTIC FIBROSIS.
>> THANK YOU VERY MUCH.
EXCITED TO BE HERE ESPECIALLY
SINCE SOME OF THE WORK REALLY
KICKED OFF MY OWN CAREER AND AS
A PEDIATRIC PULL
MONITORROLOGYIST, I WAS INSPIRED
BY THE PATIENTS I TOOK CARE OF
TO EMBARK ON A RESEARCH YOWRNY
AND I WAS ASKED TO PEEK ABOUT
IPS AIRWAY, IONOSIGHTS AND
CYSTIC FIBROSIS, SO I WILL TRY
TO INTEGRATE ALL 3 IN THE 15
MINUTE TALK.
NEXT PLEASE, AS A CLINICIAN I
REALIZE MANY OF THE LUNG
DISEASES ARE POORLY UNDERSTOOD
AND 1 OF THE REASONS IS LACK OF
EASY TO USE MODEL SYSTEMS AND
HERE'S 1, AND FOR THE AIR WAY ON
THE BASAL CELLS THAT GIVE RISE
TO MAJOR TYPES OF SECRETEATORY
CELLS AS WELL AS CELL TYPES SUCH
AS IONOSIGHTS.
SO TODAY MY FOCUS IS
CYSTIC FIBROSIS BUT I WOULD LOVE
TO BROADEN THE STUDY TO OTHER
PEDIATRIC DISEASES AS WELL.
NEXT PLEASE.
-- TO MOW AND YOU HAD -- HUMAN
AIR WAYS AND IDENTIFIED THESE
CELL TYPES THAT ARE HIGH IN CTR
BUT LESS THAN 1% OF AIRWAY
CELLS, AND THEY WERE CALLED THE
PULMONARY IONOCYTES AND IT'S
BEEN A SUBJECT OF ONGOING
RESEARCH AND I ALWAYS INCLUDE
THIS AS 1 OF MY CP PATIENTS AND
APPRECIATE THE HYPER REAMIC
MUCOSA AND THE TENACIOUS MUCUS
AND ENVIRONMENT LIKE THIS SETS
UP FOR A CHRONIC CYCLE OF
INFLAMMATION AND INFECTION,
LEADING TO LUNG STRUCTURAL
DAMAGE AND SHORTENING LIFE SPAN
EMPLOY NEXT, PLEASE.
AND AS MANY KNOW THERE'S A BREAK
THROUGH IN CF TREATMENT IN THE
FORM OF SMALL MOLECULE THAT
TARGETS THE CF MODULES, THAT
BEING SAID, GAPS STILL REMAIN,
FOR THOSE THAT DON'T HAVE
MODULATORS OR CAN'T RESPOND TO
THEM, THE FUTURE MAY BE IN GENE
CELL THERAPY AND WE ALSO NEED
PERSONALIZED DRUG AND RESPONSE
PLATFORMS SO ALL THOO NEEDS
PATIENT-SPECIFIC NONINVASIVE
HUMAN MODEL SYSTEM WHICH CAN BE
APPLIED TO CF BUT OTHER DISEASES
AS WELL.
NEXT.
SO MANY RESEARCHERS DO WORK WITH
IMMORTALIZED CELL LINES AND BUT
THEY DON'T REPRESENT ALL THE
RELEVANT CELL TYPES AND THE
PRIMARY BRORVEGGIAL HUMAN CELLS
ARE STILL THE GOLD STANDARD
BECAUSE BECAUSE THEY'RE STILL
INVASIVE THERE'S LIMITED ACCESS
FOR DIVERSE POPULATIONS AND
THEY'RE TAKEN FROM DISEASE
ENVIRONMENT WHICH MAY AFFAIRS
TEAM LEADERRER THE BEHAVIOR OF
CELLS.
NEXT, PLEASE.
SO ALL THIS TO SAY THAT INDUCED
PLURIPOTENT STEM CELL CAN BE A
PLOSM ELEMENTARY MODEL SYSTEM,
EVERYONE HERE KNOWS IPSCs, WE
HEARD A WONDERFUL TALK ABOUT IT
TAKING SOMATIC CELLS FROM
PATIENTS INTRODUCING FOREFACTORS
WE CAN DIFFERENTIATE A SOMATIC
CELL BACK TO THE EMBRYONIC STEM
CELL LIKE STATE AND IPSCs CAN
BE DIFFERENTIATED TO VARIOUS
CELL TYPES AND COUPLE IT TO THE
EDITING TECHNOLOGIES AND TARGET
BETTER YEENS.
SO IT'S PATIENT SPECIFIC,
CONTAINS HOST PROGRAMS,
NONEVASIVE, NEED TCCs OF BLOOD
AND YOU HAVE AN INEXHAUSTIBLE
RENEWAL SUPPLY.
IT'S EASE OF GENE EDITING AND
ACCESS TO DEVELOPMENTAL MINE
SOLES AND IT OFFERS A RETINAL
LOCATION SET THAT OTHER GENETICS
MAY OR MAY NOT HAVE.
I WAS FORTUNATE TO CONTRIBUTE TO
PROTOCOL BY MANY GROUPS OVER
DECADES, IT'S A STEP WISE
PROTOCOL THAT AIMS TO
RECAPITULATE IN VIVO LUNG
DEVELOPMENT.
THE FIRST 14 DAYS ARE ON 2 D, WE
MAKE IPSCs AND INHIBITING TGF
BETA AND PUT IN THE FOREGUT, AND
THEN WE MAKE LUNG PROGENITORS
WHICH ARE PURIFIED ON 3D
MATRIgE R, AND THESE ARE PURE
PIED AND PLAYED ON TO
INTERLEUKIN INTERPHASE TO,A LOW
FOR DEFERENTIATION TO MATURE AIR
WAYS AND WE CAN PRESS, WE CAN
PRAY THE ANIMATION, SO AFTER 3
WEEKS AT THE ISHT FACE, THIS IS
A BIRD'S EYE VIEW OF 1 OF THE
AIR WAY CULTURES YOU CAN SEE THE
BEATING CILIA, MEETING YOU THINK
IS A MUCUS HURRICANE AND THE
SECTION OF THE AIR WAY SHOWS A
PSEUDOSTRATIFIED EPITHELIUM THAT
MORPHOLOGICALLY VERY SIMILAR TO
A PRIMARY AIR WAY.
NEXT, PLEASE.
TO CHARACTERIZE THE POETIC
LECULAR PHENOTYPE, WE
PERFORMED SINGLE RNA SEQ, WHICH
SHOWED THAT WE GENERATED THE
RELEVANT CELL TYPES, MAININGOR
AND RARE AND THESE CELL TYPES
ARE TRANSCRIPTOMICLY SILL
SIMILAR TO AIR WAYS, ON THIS
MAP, WE HAVE 5 DIFFERENT LINES.
WE SEE GENERATIONS OF BASAL
CELLS, SECRETORY CELLS, IMMUNE
DYSFUNCTION MATURE CELLS AND
CILIATED, AND WE HAVE THE ION SO
SIGHTS THAT EXPRESS FOX 1 THAT
EXPRESS THE ISNI GENES AND THE
IONOCYTES CONSIST OF 1% OF THE
POPULATION AND WELL IS NO
CONTAMINATION FROM NONLUNG
LINEAGES USING THIS PROTOCOL.
ON THE RIGHT WE SEE STAINING OF
THE IPSCs IONOCYTESS ON THIS
AND THE LUNG FUNCTION AND AIR
WAY AND HOW THEY'RE
COMMUNICATING TO OTHER AIRWAY
CELL TYPES.L TYPES.
NEXT, PLEASE.
-- SHOWED THAT MICE DEFICIENT
IONOCYTES DEMONSTRATE VISCOSITY
AND MOST PUB LIKELY PUBLISHED,
THAT FERRETS SHOW BETTER, OTHERS
SHOWING THE IONOCYTES ARE
FUNCTIONAL BUT THEY ABSORB
CHLORIDES.
NEXT, PLEASE.
AND THEN, SO TO INTERROGATE THE
FUNCTION OF IONOCYTES TO ADD TO
THIS BODY OF LITERATURE USING
OUR SYSTEM, GIVEN IT'S ABILITY
TO DO GENE EDITING WE TEAMED UP
WITH OUR GENE EDITOR
[INDISCERNIBLE] AND KNOCKED OUT
THE FOX I1 AND DIFFERENTIATED
BOTH FOX 1 KNOCK OUT AND GENETIC
WILD-TYPE 2 AIRWAYS AND WE FOUND
THAT FOX 1 KNOCK OUT OF IPS AIR
WAY CULTURED IN THE LIQUID,
BEGAN TO CF HUMAN HAIR WAY, WITH
THE DEVELOP AM OF THESE THICK
MUCUS, IN FACT, AFTER ABOUT 3
WEEKS OF CULTURE AT INTERLEUKIN
INTERPHASE OUR TEAM HAS DAILY
WASHED OFF THE MUCUS TO SEE THE
CELLS UNDERNEATH.
AND WE ALSO SAW SIGNIFICANT
INCREASE IN MUC 5 B AND THE
CULLURES AND IT'S SIMILAR TO
WHAT'S SEEN IN THE FOX 1
KNOCKOUT MICE.
AND NOT SHOWN HERE BUT WE ALSO
SEE A SIGNIFICANT DECREASE IN
CILIATED CELLS WHICH FITS IN DAT
WITH WITH IONOCYTES AND OTHER
ORGANISMS THAT IONOCYTES MAY
COMMUNICATE TO IMPAIR CELLULAR
DIS
DYSFUNCTION.
SO WHAT IS THE FUNCTION IN
EMERGING ITS OF ION TRANSPORT SO
WE PERFORMED SHORT CIRCUIT
ANALYSIS, AND FOX 1 WILD-TYPE
AIR WAYS, SO ON THE VERY LEFT,
WE SHOW IMAGES OF THE SUCCESSFUL
KNOCKING OF THOSE IONOCYTES, THE
RED CELLS ARE ON THE WILD-TYPE
CULTURE ARE BSND POSITIVE, AND
THE KNOCK OUT CULTURES HAVE NO
IONOCYTES, SO THE FIRST CHAMBER
SHORT CIRCUIT TRACING IS
COMPARING THE FOX 1 WILD-TYPE
AND KNOCK OUT.
AND THE RED TRAITSING YOU SEE IS
THE KNOCK OUT HAS A
SIGNIFICANTLY DECREASED CHLORIDE
TRANSPORT AND CF2 R DEPENDENT AS
COMPARE TO THE BLUE WILD-TYPE
TRACING AND THERE THIS IS
SIMILAR TO THE KNOCK OUT FERIT
AND WE ALSO GENERATED A DOCKSY
PSYCHE LINE INDUCIBLE FOX 1
OVEREXPRESSING LINE IN THE KNOCK
OUT BACKGROUND, AND SO THE
SECOND WAS IN CHAMBER TRACINGOT
RIGHT, WE SHOWED THAT WEB
CONNECTED WERE ABLE TO RESTORE
THIS DEFICIENT CHLORIDE
TRANSPORT.
AND ALL THIS POINTS TO THE FACT
THAT IONON SIGHTS WERE IMPORTANT
TO AIR WAY TRANSPORT AND
FUNCTION IN A HUMAN SYSTEM.
NEXT.
WE DID A MIXING STUDY, WE
COMBINED FOX EXPRESSING IPSC
BASAL CELLS WITH A DURING
SEEDING AND THE MIXED CULTURES
WERE ALLOWED TO DIFFERENTIATE
AND THEN DOCKSY PSYCHE LYNN WAS
ADDED 5 DAYS AT IRENFACE AND TO
SAY THAT THE WILD-TYPE IONOCYTES
JUST 5% TO A CFIPS AIRWAY SEEMED
TO RESTORE THE CFTR MEDIAN
TRANSPORT AND POINTING TO A
THERAPEUTIC POTENTIAL OF THE
PULMONARY IONOCYTES.
NEXT, PLEASE.
SO BECAUSE THESE ARE SO FEW IN
NUMBERS WE NEED MONITOR THE
NUMBERS AND STUDY THE PATHWAYS
THAT LEAD TO MODULATION.
THERE'S A CRITICAL TIME AND
DIFFERENTIATION WHERE NOTCH LED
TO A SIGNIFICANT INCREASE IN ION
SIGHT MEMBERS MEMBER -- NUMBERT
IS AN ONGOING AREA OF RESEARCH
AS WELL.
NEXT, PLEASE.
AND SO WE ALSO ASKED WHETHER WE
CAN USE THE SYSTEM TO MODEL
CYSTIC FIBROSIS, THE DEFEC IN
ION AND FLUID TRANSPORT AND WE
IMENERATED -- WE HAVE PLULT PEL
IPSCs IN OUR LAB AND WE
GENERATED BOTH.
AS YOU SEE IN BLUE HAS NO
DEPENDENT CURRENT WHERE THE
CORRECTED CURRENT IN THE
CONTROLS WERE IN THE RANGE OF
PRIMARY HUMAN BRONCHIAL
EPITHELIAL CELLS AND WE'RE ALSO
ABLE TO DOWN THE BASAL CELLS AS
A 3D ORGANOID STAGE AND THE
BASAL CELLS MAINTAIN CAPABLE AS
WELL AS THIS DEFECT IN CF-ION
TRANSPORT.
NEXT PLEASE.
SO IN THE LAST FEW MINUTES,
ANOTHER GAP WE MENTION SIDE WE
NEED PATHOGEN IS DRUG RESPONSE,
NEXT, PLEASE, SO DURING THE
PANDEMIC AS EVERYBODY DID, WE
PIVOTED OUR PLATFORM TO MODELS
SARS-COV-2, AND TEAMED UP WITH
VIROLOGISTS FROM THE NIAID WITH
CULTURES FROM SARS-COV-2 WE
FOUND OURSELVES PERMISSIVE TO
THE VIRUS AND ON THE LEFT IS THE
VOLCAN O PLOT SHOWING THE IPS C
AIR WAYS AND LIKE PRIMARY AIR
WAYS THESE EXHIBIT VERY ROBUST
TYPE 1 AND TYPE 3 INTERFERON
RESPONSE AS WELL AS PLEURAL
INFLAMMATORY RESPONSE AND TO THE
RIGHT WE ALSO LOOKED AT THE
FEASIBILITY OF THE THIS PLATFORM
TO SCREEN FOR COVID-19.
THERAPEUTIC COMPOUNDS AND HERE
I'M SHOWING REMDESAVIR BUT ALSO
TESTED SEVERAL OTHER COMPOUNDS
IN THIS FASHION AS WELL.
NEXT.
AND BECAUSE THE IPSC SYSTEM
OFFERS THIS REDUCTIONIST
PERSONALIZED MODEL SYSTEM AND
WHEN COUPLED GENE EDITING ALLOW
US TO TEASE OUT THE EFFECT OF A
SPECIFIC GENE ON PATHOGEN
RESPONSE.
IN THIS CASE WE INFECTED CF AND
THE CFTR, WITH THE SARS-COV-2
AND WE FOUND THAT CF EPITHELIUM
HAD A SIGNIFICANTLY HIGHER VIRAL
BURDEN WITH THE EPITHELIUM WITH
DISORGANIZED SEALIA AND THE
GREEN HERE IS SARS-COV-2 AND
PROTEIN AND THEN THE RED IS
[INDISCERNIBLE] CELLS SO THE CF
LOOKS MUCH SICKER BUT THEN
INTERESTING, WHEN WE TREATED THE
CF AIRWAYS WITH THE MODULATOR IN
THE COMBINATION, WE FOUND THAT
THE CFTMODDULATOR HAD
SIGNIFICANT REDUCTION OF
INFECTION AND A MUCH BETTER
LOOKING AIRWAY EPITHELIUM.
NEXT, PLEASE.
SO WE WANTED TO MAKE SURE IT'S
NOT A SARS, JUST A ONLY
PHENOMENON.
WE WANT TO LOOK AT OTHER VIRUSES
AS WELL AND FOR THIS WE TEAMED
UP WITH [INDISCERNIBLE].
AND WE INFECTED OUR IPSCs WITH
H1N1 AND SO THAT AIRWAY HAD
INCREASE INFECTION COMPARED TO
CANNED WHAT AIRWAY AND HERE THE
GREEN CELLS ARE IN NUCLEOTIDES
CLARPROTEIN AND WHEN WE HAVE THE
MODULOR TRI CASTA THERE WAS A
SIGNIFICANT REDUCTION IN
INFECTION.
NEXT.
NEXT, SO IN SUMMARY, I THINK I
HOPE TO HAVE SHOWN YOU THAT WE
HAVE ESTABLISHED IPSC AIRWAY
THAT CAN BE DIFFERENTIATED INTO
COMMON AND RARE AIRWAY CELL
TYPES TO STUDY THEIR BIOLOGY AND
WE CF AND WERE ABLE TO CAPTURE
THE DEFECT IN CF ION TRANSPORT.
WE HAVE GENERATED IONOCYTES, AND
FASCINATING CELL TYPES WHICH
SEEMS TO BE A KEY REGULATOR FOR
HOMEOSTASIS IF YOU KNOCK IT OUT
AND THEN WHEN YOU OVEREXPRESS
IT, WE CAN USE ITS PLATFORM FOR
PLATFORM INFECTION AND T-CELL
DISEASE EPITHELIAL RESPONSE TO
PATHOGENSAs WELL AS RESPONSE
TO DRUGS.
AND TO SAY THAT AGAIN, THE
FUTURE DIRECTION IS TO EXPAND
THIS PLATFORM BEYOND
CYSTIC FIBROSIS AS WELL.
SO THE LAST FEW SLIDES, I WOULD
LIKE TO TALK ABOUT GAPS.
NEXT PLEASE, AND IN TERMS OF
BOTH IONOCYTES BIOLOGY AND
IPSCSYSTEMS, IN TERMS OF
IONOCYTES WE STILL NEED TO
UNDERSTAND THE DIFFERENT
SUBTYPES, JUST IONOCYTES INTO
OTHER TISSUES HAVE DISTRIBUTION,
LOCALIZATION, AND THE FUNCTIONS
MAY BE DIFFERENT.
NEED TO KNOW A SIGNALING PATHWAY
THAT LEAD TO DIFREBTIATION,
STUDY THEIR ROW IN AIRWAY
HOMEOSTASIS AND ALSO TON IF
THEIR FUNCTION DIFFERS IN
DIFFERENT AIRWAY DISEASES THAT
HAVE DIFFERENT AREAS OF
INFLAMMATION AND HOW OUR
ENVIRONMENT INFLUENCES THAT
FUNCTION.
AND AN INTRIGUING QUESTION IS
WHETHER OR NOT ION SO --
IONOCYTES CAN BE TARGETED FOR
GENE THERAPY.
ACTUALLY CAN YOU GO TO THE NEXT
SLIDE.
OKAY, IN TERMS OF IPSCSYSTEMS,
TO THE BEST OF MY OWN KNOWLEDGE,
THE GAPS IS THAT IPSC TISSUES
FOR THE MOST PART ARE STILL MORE
IMMATURE, AND WE REALLY DO NEED
TO TAKE AN ACCOUNT THE MATRIX,
MECHANICAL PROPERTIES IMMUNE
CELLS AND VASCULAR CELLS.
WE NEED TO STUDY HOW RESPONSE TO
STRESSORS AND SCALABILITY, I
THINK IS A BIG DIESEL, IT'S
EXPENSIVE AND LABORIOUS AND THE
METHOD NEEDS TO BE MORE
STANDARDIZED.
AND SO, NEXT, I THINK THIS IS MY
LAST SLIDE, SO THE KEY RESEARCH
QUESTIONS IS THE OPTIMAL
CONDITIONS TO GET THE SPECIFIC
LUNG TYPES AND HOW DO EACH
GENETIC VARIATIONS EFFECT IPSC
LUNG CELLS AND THEN, CAN WE
MODEL THE DIFFERENT DISEASES AND
ALSO RESPONSE TO ENVIRONMENT AND
HOW THE CELL TYPE RESPONDS TO
ENVIRONMENT WITH THAT I LAST
SLIDE IS TO ACKNOWLEDGE THE
WONDERFUL PEOPLE I GET TO WORK
WITH EVERY DAY AND VERY NEW
WOBDERFUL LAB MEMBERS AND
AMAZING COMMUNITY WHO I GET TO
COLLABORATE WITH.
THANK YOU VERY MUCH.
THIS, THANK YOU SO MUCH
DR. WANG.
OUR NEXT SPEAKER IS DR. JOHN ENG
ELHARDT, WITH THE PROFESSOR OF
DEPARTMENT OF ANATOMY AND CELL
BIOLOGY, ROY J. CARVER CHAIR OF
MOLECULAR MEDICINE DIRECTOR
CENTER FOR GENE THERAPY CARVER
COLLEGE OF MEDICINE, UNIVERSITY
OF IOWA, HE STUDIES VECTORS TO
MODEL SYSTEMS AND TODAY ME WILL
SPEAK TO US ABOUT ASSESSING LUNG
INJURY AND REPAIR USING
TRANSIMENNIC FERRET MODELS.
THE FLOOR IS YOURS.
NTHANK YOU.
NEXT SLIDE.
THIS IS A VERY WE'VE HAD SOME
WONDERFUL PRESENTATIONS EARLIER
TODAY ABOUT DIFFERENCES IN,A NAT
ME AND CELL BIOLOGY BETWEEN
HUMANS AND RODENTS IN THE LUNG
AND AIR WAY.
AND THIS IS A VERY SUPERFICIAL
VIEW OF THE DIFFERENCES I WILL
TALK ABOUT TODAY.
ONE INCLUDES THE ABUNDANCE OF
GLANDS IF YOU LOOK ON THE LEFT
IN LARGER SPECIES IN HUMANS AND
BASAL CELLS ARE DISTRIBUTED
THROUGH CARTILAGE NOWS AIRWAYS
AND THE TYPE OF SECRETORY CELLS
ARE DIFFERENT THAN THAT OF
RODENTS AS WELL.
THESE ARE REASONS, NEXT, THAT
CYSTIC FIBROSIS OR CFTR KNOCK
OUT IN PIGS AND FERRETS DEVELOP
OBSTRUCTIVE LUNG DISEASE, MICE
DON'T AND THERE'S HEAVY GLAND
INVOLVEMENT IN THAT GENETIC
DISEASE.
NEXT.
AS SHOWNOT PREVIOUS SLIDES,
THERE WERE DISTAL AIRWAYS, THE
BRONCHIALES THAT DISOANT EXIST
WITHIN MICE BUT EXIST WITHIN
FERRETS AND HUMANS THAT HAS
NEXT, SPECIALIZED STEM CELL
POPULATIONS THAT WE'VE HEARD
ABOUT.
NEXT.
THAT WE'VE HEARD ABOUT
PREVIOUSLY INCLUDING RAS,
TERMINAL RESPIRATORY AND
AT-CELLS.
NEXT.
SO FOR QUITE A WHILE, I'VE
STUDIED THE STEM CELL BIOLOGY OF
SUBMUCOSAL GLANDS IN THE CONTEXT
OF CYSTIC FIBROSIS, WE DEMON
TRAITED USING MICE IN THE 2 CRE
DRIVERS MARKED HERE THAT THE
STEM CELL POPULATION IS RESERVE
STEM CELL THAT CAN REPOPULATE
STEM CELLS WITHIN THE GLAND DUCT
OR SURFACE AIR AIRWAY EPITHELIUM
ALONG WITH TATAWHO SAW SIMILAR
RESULTS.
NEXT SLIDE.
SO WE GENERATED THE SAME TYPE OF
LINEAGE TRACING FERRETS, AS WE
HAD STUDIED IN MICE AND WHAT YOU
CAN SEE HERE IS THIS IS USING AN
IREZ INSERTION OF THE ACTED 2
GENE.
AND AT HOMEOSTASIS, WHEN THESE
ANIMALS ARE TRACE WIDE TAMOXIFEN
ON THE LEFT AND SMOOTH MUSCLE ON
THE INTERMULL MONITORARY ON THE
AIRWAYS.
NEXT SLIDE.
AND INDEED IF WE 0 THESE DOUBLE
TRANSGENIC FERRETS THAT HAVE M
TMG REPORTER WITH TISSUE AND
MACROPHAGES OX FEN 4 WEEKS LATER
OR WE LABELED WITH TISSUE AND
MACROPHAGES OX FEN AND LABELED
THEM LATER, HARVEST, 1 SEES A
SIMILAR PATTERN OF DISTRIBUTION
THAT 1 WOULD SEE ONLY IN THE
PROXIMAL TRACHEA OF MICE IN
WHICH MILE EPITHELIAL CELLS
EMERGE FROM THE GLAND DUCTS,
NEXT, AND DIFFERENTIATE INTO
BOTH DUCTAL CELLS MARKED BY THE
ARROW THERE.
GLANDULAR TUBEULAR CELLS
INCLUDING SECRETORY AND SERA
CELLS AND ON THE AIR WAY SURFACE
ACCIDENT BASAL CELLS MARKED BY
P63 AND SECRETORY CELLS MARKED
BY MUC5 AC.
NEXT SLIDE.
AND WE WERE INTRIGUED BY THE
DISTAL AIRWAY DIFFERENCES
BETWEEN RODENTS AND FERRETS IN
THESE LINEAGE TRACING MODELS.
ONE CAN SEE ON THE LEFT,
LEFT-HAND SIDE, THE BRONCHIAL
ALVEOLAR JUNCTION WE JUST HEARD
QUITE A BIT ABOUT FROM DR. ZHOU,
THE SMOOTH MUSCLES EXIST AT THIS
TERMINAL REGION AS SPARSE
INDIVIDUAL CELLS AS COMPARED TO
THE RIGHT WITH THE RESPIRATORY
BRANCHIAL OR RAS, FOR THE
SECRETOR I SEMESTER CELLS, HAVE
SMOOTH MUSCLE BUNDLES JUST BELOW
THE SPECIALIZED CLUB CELL THAT
EXIST IN THE RESPIRATORY CELL.
NEXT SLIDE.
THIS REGION IS QUITE UNIQUE, IF
1 LOOKS AT THE AREA MARKED BY AN
ARROW, 1 SEES THAT SECRETORY
EXPRESSING CLUB CELLS DECLINED
SIGNIFICANTLY WITH CILIATED
CELLS MARKED BY ACELLALATED TUBE
LYNN AND AS DR. BASIL SHAZ SHOWN
AND TATA'S GROUP THAT THIS
SPECIALIZED 3 A2 EXPRESSING CELL
EMERGES AND DOMINATES AND IN THE
TERMINAL BRONCHIAL AND SHOWN ON
THE LEFT, VERY LEFT, RESPIRATORY
BRONCHIALS, RB.
NEXT SLIDE EMPLOY NSO WE
HYPOTHESIZED THAT SPECIES
SPECIFIC DIFFERENCES IN THIS
PARTICULAR REGION AND IN ANATOMY
AND CELL BIOLOGY IN THIS
RESPIRATORY ZONE MAY CONTRIBUTE
TEHUMAN ASSPECS OF FIBROSIS
FOLLOWING ACUTE LUNG INJURY, SO
WE DEVELOPED AIBLY O MICEIN
INJURY MODEL IN THE FERRET
THAT'S SHOWN IN THE TOP.
THREE TYPES INJURED WITH
BLEOMYCIN, AND THEN HARVESTED
ANIMALS SUBSEQUENTLY OUT TO 12
WEEKS AND YOU CAN SEE ON THE
LEFT WITH BOTH THE RADIO GRAPHS,
CT IMAGES AS WELL AS HISTOLOGY,
SIGNIFICANT FIBROSIS AND
CONSOLIDATION WITHIN THE LUNG.
FUNCTIONALLY, THE CAPACITY
DECLINES AS SHOWING THE CENTER
AND THIS CORRELATES WITH THE
SCORE WHICH IS AN INDEX OF
FIBROSIS, AND YOU CAN SEE THESE
FIBROTIC FOCI, AND STARTING TO
FORM IN THE LOWER RIGHT.
NEXT SLIDE.
SO WE PERFORMED SINGLE YOU
THIS CLEAR SEQUENCING AND
COLLABORATION WITH
[INDISCERNIBLE] IN IMI
BIOSCIENCES AND ON THE LEFT-HAND
SIDE YOU CAN SEE THE UMAP WITH
THE TYPICAL CELL ASSIGNMENTS BUT
IMPORTANTLY INCLUSION OF BASIL
LIKE CELLS ON THE RIGHT, ONCE
THE SIGNIFICANT TRANSCRIPTIONAL
CHANGES IN THIS UMAP WHEN 1
MARKS THE SALINE, 1 SEALS THE
CELL TYPES THAT ARE SHOWN ON THE
LEFT, AGAIN, SIGNIFICANT
TRANSCRIPTIONAL CHANGES AND
NEXT, IF 1 LOOKS AT THE MAGENTA
ARROW HEAD SIGNIFICANT CHANGES
IN THE DISTRIBUTION OF A NUMBER
OF POPULATIONS INCLUDING
TRANSITIONAL AT2 CELLS IN
BASALOID LIKE CELLS.
IMPORTANTLY PHENOTYPICALLY, THIS
TP63 POSITIVE KRT NEGATIVE KRT 7
POSITIVE CELL MARKED BY THE
YELLOW ARROWS EXIST WITHIN THE
FERRET, TISSUE FOLLOWING
BLEOMYCIN, THIS IS A POPULATION
WE HEARD ABOUT THAT IS NOT FOUND
IN MOUSE.
NEXT SLIDE.
AND IF 1 PROJECTS THE
TRAJECTORIES IN PSEUDOTIME SHOWN
HERE JUST FOR THE AT2, AT1 LIKE
CELLS GROUPED TOGETHER, 1 SEES 2
TRAJECTORIES FROM THE RIGHT
PANEL THAT EMERGE THROUGH
THROUGH TRANSITIONAL AT2 CELLS
TO BASAL 1 AND AND NEXT, THESE
ARE JUST SPECIFIC MARKERS,
MARKING THESE POPULATIONS.
NEXT.
AND 1 CAN SEE, THIS BASALOID
POPULATION AT THE TOP OF THESE
KERATIN, AND ABERRANTLY KERATIN
EXPRESSING CELLS, UNFORT MATILY
KERT EXPRESSION IS NOT SHOW SO
WE CAN BE THE SO A UMAP OF THAT.
NEXT.
AND THERE ARE INTERESTING
ANATOMICAL SIMILARITIES TO
PROGRESSION OF FIBROSIS WITHIN
HUMANS AND IPF, WHERE
REGIONALLY, THE WORST AREAS OF
DISEASE ARE AT THE SURFACE, THE
PLURAL SURFACE OR PERIPHERY OF
THE LUNG, HERE WE'RE SHOWING THE
AREA MARKED IN A, STAINED FOR
KRT 7, NEXT, VERY LITTLE
EXPRESSION HOW THE KRTs CAN
PROGRESSIVELY BE AT THIS
POSITION MARKED B, NEXT, YOU SEE
INCREASING EMERGENCE OF KRT 17
AND 5 EXPRESSION.
NEXT.
AS 1 PROGRESSING TO THE EXTERIOR
OF THE LOBE, HERE MARKED BY
ARROWS ARE AGAIN, THIS ABERRANT
KRT 5 NEGATIVE KRT 7 POSITIVE,
KR17 POSITIVE CELL.
NEXT.
THIS IS JUST ANOTHER EXAMPLE OF
THE SIGNIFICANT TYPE OF
PROXIMALLIZATION THAT'S
OCCURRING WITHIN THE ALVEOLAR
SPACE LIKELY THROUGH NICHE
COMPETITION TO REGENERATE THE
ALVERMEN INFECTED O LIE AND THE
FIBROSIS THAT INDUCES EXPANSION
OF THE BASAL AND BASALOID
POPULATIONS, MARKED BY ARROWS
AGAIN IN THE UPPER RIGHT ARE
TP63, KRTR 7, KRT 5 XDAPI CELLS
MARKED IN GREEN THAT EXPRESS KRT
7.
NEXT.
IF 1 CLUSTER IS JUST THE
MESENCHYMAL CELL TYPES WITHIN
THE EPITHELIAL CHALLENGED LUNG,
1 SEES HERE, WE HAVE ALVEOLAR
FIBROBLAST TYPE 1 AND 2, VARIOUS
SMOOTH MUSCLE GROUPS, AND
PERISIGHTS, 1 SEES THE
EXPRESSION ON THE RIGHT AS
FAIRLY SIMILAR TO WHAT'S BEEN
PUBLISHED WITHIN HUMAN -- HUMAN
LUNG MAP ON THE TOP AND THAT
FERIT SEQUENCE ON THE BOTTOM,
NEXT.
IF 1 CLUSTERS OR 1 MAPS THE
SALINE VERSUS THE BLEOMYCIN
ANIMALS, THE TRANSCRIPTIONAL
CHANGES WITHIN THE SMOOTH MUSCLE
COMPARTMENT ON THE LOWER LEFT AS
WELL AS ON THE ALVEOLAR
FIBROBLASTS TYPE 1 COMPARING ON
THE RIGHT.
NEXT SLIDE.
SO WE HYPOTHESIZE THAT
RESPIRATORY SMOOTH MUSCLE
CONTRIBUTES TO THE FIBROBLASTS
IN MOUSE AND FERIT AND WE
COMPARED IT NEXT EMPLOY WHAT I'M
SHOWING HERE IS AN ENACTED ROSA
TG MOUSE THAT'S BEEN BLEOMYCIN
INJURED AND THEN TAMOXIFEN
LABELED ONCE IT'S OVER AND YOU
SLEEP APNEA AND OBESITYY HOW IT
TRACE, AND THE MY O FIBROBLAST
EXPANDING.
IF 1 LABELS PRIOR TO BLEOMYCIN
INJURY AND WASHES OUT 1 SEES
VERY LITTLE OVERLAP, OR VERY
LITTLE LABELING REALLY JUST THE
VESSELS IF 1 LOOKS IN THE TOP
RIGHT THAT ARE LABELING WITH TO
POST BLEOMYCIN INJURY.
NEXT.
THIS HAS BEEN SHOWN IN A NUMBER
OF PUBLICATIONS THAT ACTA 2
LINEAGE DO NOT CONTRIBUTE TO MY
O FIBROLASTS IN MICE FOLLOWING
BLEOMYCIN AND MOUSE.
NEXT SLIDE, THIS COMPARES TO
THIS TO FERRET HOWEVER, IT'S ARE
A VERY DIFFERENT OUTCOME.
ON THE LEFT-HAND SIDE, IS
LABELING AFTER BLEOMYCIN INJURY.
YOU WOULD EXPECT, YOU CAN SEE
SIGNIFICANTLY AS WELL AS
STAINING BUT IMPORTANTLY ON THE
RIGHT, IF 1 LABELING HOMEOSTASIS
WASHES OUT, THEN BLEOMYCIN
INJURIES, 1 SEES IF YOU FOCUS ON
THE MOST RIGHT PANELS WHICH ARE
ENLARGEMENTS OF THE YELLOW
ARROWSOT LEFT, 1 SEES
SIGNIFICANT OVERLAP IN THE
[INDISCERNIBLE] STAINING AND
REGIONS DISINTEGRATING
BRONCHIALS WITH ACTA 2 LINEAGE
TRACING.
NEXT.
NOW THERE'S SIGNATURES EXPANSION
OF KRT POSITIVE CELLS AROUND
LINEAGE TRACED [INDISCERNIBLE]
MY O FIBROBLASTS THAT ARE WITHIN
THE PATHWAY GIVES RESERVING MA,
COMPARING NEXT THIS ENLARGED
REGION RIGHT HERE, THAT THE
SIGNIFICANT AMOUNT OF OVERLAP
BETWEEN THE ACTA 2 AND TRACE AND
ALPHA [INDISCERNIBLE]
EXPRESSION, THESE ANIMALS WERE
LABELED WITH TISSUE AND
MACROPHAGES OX FEN FIRST AND
THEN CHASED AND INJURED WITH
BLEOMYCIN AND IN THE YELLOW
DOTTED BOX REGION, 1 CAN SEE THE
CLUSTERS AROUND THESE MY O
FIBROBLASTS THAT HAVE EMERGED
LIKELY FROM THE BRONCHIALS BUT
WE CAN'T RULE OUT THE VASCULAR
SMOOTH MUSCLE AT THIS POINT, THE
OTHER INTERESTING THING IS,
NEXT, IF 1 LOOKS AT THE RESIDUAL
SMOOTH MUSCLE OR THE SMOOTH
MUSCLE AROUND THE TERMINAL
BRANCHIALS ALOS ANGELES AS
RESPIRATORY WRONKIALS, 1 SEES
THAT THE TRACE IS LOST, BUT ACTA
2 EXPRESSION IS RETAINED
SUGGESTING THAT SOMETHING THAT
THE SMOOTH MUSCLES AROUND THAT
REGION OF THE AIRWAY ARE BEING
REPLACED BY SOME OTHER
PROGENITOR THAT DOESN'T EXPRESS
ACTA 2.
NEXT SLIDE.
SO IN CONCLUSION, WE FEEL THERE
ARE UNIQUE ASPECTS OF HUMAN LUNG
ANATOMY AND CELL BIOLOGY AND THE
RESPIRATORY ZONE THAT COULD BE
BETTER MODELED WITH WITHIN THE
FERRET AND WE'RE EXCITED TO
APPLY THE TRANSGENIC FERRETS AND
TECHNOLOGIES TO GENERATE THESE
ANIMALS, TO TACKET THESE
QUESTIONS.
AND NEXT.
AND MOVING ON SOME OF THE
EXCELLENT WORK OF DR. BIN ZHOU,
WE GENERATED AN INTERSECTIONAL
REPORTER FERRET AND WORKING ON
THE CRE-DRIVER FERRETS TO ANSWER
THE DIFFICULT QUESTIONS ABOUT
THE MESENCHYMAL ORIGINS OF MIGHT
BE O FIBROBLASTS AND WHAT
THEY'RE DOING, ARE SOME OF THEM
BENEFICIAL, SOME DETRIMENTAL TO
THE FIBROTIC RESPONSE?
NEXT.
AND LASTLY, TRANSGENIC OR FERRET
TRANSGENIC FETE MAPPING MODELS
COUPLED WITH CONDITIONAL GENETIC
APPROACHES MAY ENHANCE
TRANSLATABLE OF BASIC RESEARCH
AND THUS THERAPIES TO HUMANS.
NEXT SLIDE.
I WOULD LIKE TO THANK THOSE IN
MY GROUP, WORKING WITH FERITS
AND TRANSGENIC FERRETS.
IT'S A MAJOR EFFORT.
THERE ARE MAJOR COLLABORATORS
INCLUDING VISHWARAJ, THE
MORESESY LAB, TATA'S LAB AND
OTHERS LISTED HERE .
THANK YOU.
>> THANK YOU DR. ENG ELHARDT,
NOW WE CAN CAN BEGIN THE
DISCUSSION.
SOME POINTS OF DISCUSSION, WE
ARE HERE TO TALK ABOUT WHAT THAT
WE CAN DO WITH THIS TECHNOLOGY
WHAT ARE THE LIMITATIONS AND HOW
WE CAN IMPROVE ON THEM.
NEXT, PLEASE.
NTO START WITH, YOU SHOWED THESE
BEAUTIFUL, THE EMBRYO ALZHEIMER
BEHAVES SO WELL INVITRO.
AND THE ADULT IS ALWAYS
CHALLENGING EMPLOY AND MY
QUESTION, I HAVE 2 LEVELS OF
QUESTION 1 AND TATA SUPPOSE
ABOUT THIS ALSO BUT CAN WE GET
SOME SELF-ORGANIZATION OUT OF
ADULT STRUCTURES OR DO WOE HAVE
TO LIKE POSITION THEM PERFECTLY?
AND THEN SECOND COMPONENT OF
THAT IS, WE OFTEN THINK ABOUT,
YOU KNOW THE BABY AND THE ADULT
BUT THERE'S AN INTERMEDIATE
STAGE IN PEDIATRICS LIKE THE
TEENAGER AND IS THERE A TIME
WHEN YOU THINK THERE'S STILL
SELF-ORGANIZATIONAL PROPERTIES
IN 3 ADULT LUNGS THAT YOU COULD
EXPLOIT INVITRO?
>> YEAH, THANKS SO MUCH FOR
THOSE WONDERFUL QUESTIONS.
I THINK ORGANOID OR IPSC DERIVED
[INDISCERNIBLE] BOTTLENECK FOR
THE ENTIRE FIELD.
AS YOU MENTIONED WITH A CURRENT
MODEL IT'S BETTER TO STUDY
DEVELOPMENTAL PROCESS, BUT
NONAPOPTOTIC MODEL THE
COMPLEXITY AND ADOPT LUNGS
ESPECIALLY FOR SCALE UP, OUR
CURRENT TEENY TINY SYSTEM, IT
WILL REQUIRE A LOT OF EFFORT,
BECAUSE YOU KNOW THAT LUNG IS SO
MANY YEARS OF HUMAN DEVELOPMENT
WHEREAS OUR CURRENT SYSTEM IS
BASICALLY AT MOST 3 MONTHS OF
DIFFERENTIATION.
SO I THINK 1 EASY METHOD TO
UNDERSTAND SOME QUESTIONS, IN
THAT LUNG PHYSIOLOGY OR
PATHOPHYSIOLOGY AND TO LEVERAGE
ENGINEER KAFENDAR--ALEXANDER
OLD, SO WHAT WE COULD DO IS TO
DEFERENTIATE THOSE CELLS AND
THEN WE SEE THEM ON THE RIGHT
STRUCTURE TO STUDY SOME
INTERACTIONS, IN A REDUCED
DIMENSION.
THAT'S OUR CURRENT WAY OF USING
THESE ORGANOIDS TO UNDERSTAND,
YOU KNOW SOME OF THE ADULT HUMAN
LUNG QUESTIONS.
AND FOR PEDIATRIC LUNGS, I THINK
IT FALLS IN THE SAME, I GUESS
AREA OF DISCUSSION, HOW FAR WE
CAN JUST PUSH THE CURRENT SYSTEM
BUT I THINK SOME OF THE LUNG
RESPONSES TO INJURY RECAPITULATE
THE DEVELOPMENTAL PROCESS, SO IN
THAT REGARDS WE MIGHT BE ABLE TO
USE THIS PRENATAL SYSTEM TO
UNDERSTAND LONG REGENERATION AND
REPAIR IN RESPONSE TO INJURY,.
>> YOU KNOW, AREN'T YOU
BUILDING, AREN'T YOU ADDING MORE
ALVERMEN INFECTED O LIE WHEN
YOU'RE 2 YEARS OLD AND 4
YEAR-OLDS, AND ISN'T THERE SOME
NASCENT CAPACITY FOR
SELF-ORGANIZATION THAT YOU COULD
HARVEST AT THOSE STAGES?
>> I THINK SO.
I THINK SO, THE ALVEOLAR
DEVELOPINGY POSTINATEALLY.
>> SO MAYBE THOSE ARE GOOD
ENOUGH TO STUDY ADULT DISEASE.
MAYBE YOU DON'T HAVE TO START
WITH ADULT CELLS IF THAT'S
DOABLE.
>> YEAH, ABSOLUTELY.
THOSE ARE THE QUESTIONS
ADDRESSABLE USING THE IS.
YEAH, QUITE DEPENDENT ON WHAT
OUR LIST OF QUESTIONS WE'RE
ASKING USING THIS ORGANOIDS AND
THEY'RE SERVING AS A
COMPLIMENTARY PLATFORM TO FERIT
OR MOUSE OR EVEN LARGER ANIMALS.
>> I THINK [INDISCERNIBLE] HAD A
QUESTION.
>> I DO HAVE A QUESTION, I DON'T
KNOW WHETHER MY HAND OR -- CAN
YOU HEAR ME?
>> YEAH.
WE CAN HEAR YOU, YOU CAN BRING
UP YOUR QUESTION.
>> THANK YOU VERY MUCH.
I WONDERED DR. ZHOU, VERY -- ALL
PRESENTATIONS WERE VERY
WONDERFUL, DR. ZHOU, I HAVE IA
QUESTION FOR YOU, IT LOOKS LIKE
HAVING THE CLUB CELLS, HAVING
THE CAPABILITY OF BECOMING
ESSENTIALLY TYPE 2 CELLS AND
TYPE 1 CELLS AND LIKE THE OTHER
CELLS AS WELL, BECOMES SO
FLEXIBLE IN PLASTICITY TO
DIFFERENTIATE IN CELL TYPES, IS
IT POSSIBLE THAT DEPENDING ON
INJURY SAY FOR EXAMPLE, YOU DID
BLEOMYCIN, SAY IF YOU HAD
ANOTHER INJURY, LET'S SAY OR
OTHER TYPES OF CHRONIC INJURY
MODELS, WOULD YOU EXPECT OTHER
CELL TYPES THAT DOMINATE THE
DIFFERENTIATION PATHWAY GOING
INTO -- DEPENDING ON WHAT TYPE
OF INJURY AND HOW THE CELL
RESPONDS TO THE INJURY, THAT
PLASTICITY IS IT SOMETHING THAT
COULD BE-SOMETHING TO LOOK INTO?
>> YEAH.
THANK YOU FOR YOUR QUESTION.
YEAH, IT'S POSSIBLE THAT IF YOU
GIVE A DIFFERENT INJURY, OTHER
RESIDUE REMAINING PROGENITORS
CAN RESPOND CAN COMPENSATE FOR
THE LOSS OF THE EPITHELIAL CELLS
THAT ARE INJURED BY THE SPECIAL
CONDITIONS SO IN OUR CASES, WE
INDUCE AN ARTIFICIAL MODEL FOR
SEVERE INJURY ON THE ALVEOLAR
AT2 CELLS.
WE USE P21 OR WE CAN ALSO USE
DTR SYSTEM TO ABLATE A LARGE
PORTION OF AT2 CELLS SO THAT
OVEREXPRESSION OF P21 IMPAIRED
THEIR PROLIFERATION POTENTIAL,
SO IN THIS CASE, SPECIAL CASE,
ARTIFICIAL CASE, MODEL, THE CLUB
CELLS PLASTICITY TO REGENERATE
VIRTUALLY ALL AT2 CELLS IN SUCH
CONDITIONS.
WE SHOWED THIS BECAUSE WE CAN
SEE IT.
WE CAN DISTINGUISH IT, FROM
BASCs, CLUB CELLS AND AT2
CELLS, USING FEWER RECOMBIN ACES
THAT WE CAN TRACE 3 POPULATIONS
SIMULTANEOUSLY WITH 3 DISTINCT
COLORS, FOR INSTANCE THE
REPORTERS AND WE CAN SEE IT, BY
THIS MODEL.
I AGREE WITH YOU THAT IF WE
ABLATE OR INJURED THE CLUB
CELLS, LIKE USING
[INDISCERNIBLE] INJURY MODEL, WE
IS SEE THAT AS A BASC CELLS, THE
BASC CELLS CAN MIGRATE INTO
BRONCHIALES AND REGENERATE CLUB
CELLS IN OUR PREVIOUS STUDIES,
AND USING SPECIFIC LINEAGE
TRACING OF BASCs SO IT LARGELY
DEPENDS ON WHICH MODEL, YOU USE
FOR STUDIES THE LUNG
REGENERATION.
THANK YOU FOR THE QUESTION.
>> THANK YOU.
NTHAT WOULD INDICATE, I MEAN
JUST AS A GENERAL DISCUSSION,
YOU KNOW, ALTHOUGH THE MODELS
ARE WONDERFUL, TRYING TO CAPTURE
THIS COMPLEXITY OF THE LUNG, IN
TERMS OF BEING ABLE TO BE SO, SO
CONDUCIVE OR SO ESSENTIALLY
ADJUSTABLE TO WHATEVER IT GETS
AS AN INJURY, THAT'S SOMETHING
THAT WE SHOULD PROARP --
PROBABLY THINK ABOUT MORE.
OKAY, IS THE QUESTION FOR ME?
>> YEAH.
HI BIN, GOOD TO SEE YOU IF I MAY
ASK IN THE SAME LINE, SO IN SOME
INJURY MODELS, WE SEE THE
MIGRATION OF [INDISCERNIBLE]
CELLS IN CONTRIBUTION TO YOU
KNOW ALVEOLAR, THEY MOVE TO
ALVEOLAR DAMAGED REGIONS, RIGHT?
SO I WAS WONDERING DO YOU THINK
SIMILARLY, THE NOTCH SIGNALING
IS ALSO PLAYING YOUR ROLE IN
MODELIZATION OF THIS VERSION IN
THE MODEL, NOT IN A GAIN OF
FUNCTION MODEL BUT JUDGE UOF THE
IN A NONINJURY REPAIR.
IF YES, HOW IS THE NOTCH
SIGNALING ACTUALLY WORKING BY
NEIGHBORING CELLS INTERACTING
AND NOTCH LIGAND FROM 1 CELL TO
ACTIVATING OTHER CELL.
SO I'M WONDERING HOW THE -- NYOU
MEAN THE CLUB CONTRIBUTION TO
AT2 CELL REGULATION BY NOTCH IN.
>> THAT'S RIGHT.
>> YEAH, IT'S POSSIBLE THAT
[INDISCERNIBLE] CELLS MIGHT
PROVIDE THE NOTCH SIGNALING TO
THE CLUB CELLS, THAT PROMOTED
THEIR DIFFERENTIATION INTO AT2
CELLS AFTER THE YOUR.
SO WE -- 1 OF MY STUDENTS ARE
TRYING TO PURSUE IT HERE FOR
MANIPULATION OR THE NEIGHBORING
CELLS OR CLUB CELLS IF THEY CAN
HAVE AN IMPACT ON THEM.
DIFFERENTIATION, TO THE IT2
CELLS AND WORKING ON THAT, WE
DEVELOP SOME AND INTRACELLULAR
GENETIC APPROACH USING SIGNAL
NOTCH TO MANIPULATE THE NEIGHBOR
OR CELLS TARGETED CELLS TO SEE
IF WE'RE -- THEY HAVE SUPPORT
PROVIDE A NICHE OR SUPPORTIVE
SIGNALS TO THE SURROUNDING,
YEAH, CELLS, WE ARE STILL
WORKING ON THAT.
THANKS FOR THE QUESTION.
>> THANK YOU.
>> I CAN READ OUT 1 OF THE
QUESTIONS ASKED IN CHAT TO HALL
CHAPMAN TO THE BIN ZHOU, THE
CAPACITY FOR CLUB CELLS IN THE
GRAPH, HAVE YOU TRIED TO DOUBLE
LABEL THIS POPULATION AND IF SO,
WHAT EXTENT OF CC10 TRACE IS
[INDISCERNIBLE] TOWARDS THIS
NTHANKS FOR THE COMPLEMENTS AND
QUESTION.
YEAH, WE HAVEN'T DONE ANY
INGRAFTMENT AND TRANSPLANTATION
EXPERIMENTED BUT IT IS IF YOU
HAVE MODULATION THROUGH THE CLUB
CELLS WE CAN DO THIS TRACING OF
SUBPOPULATION OF CLUB CELLS IN
VIVO OR IN TRANSPLANTATION
ASSAY.
I THINK RECENTLY THEY STARTED TO
SHOW COLLABORATION WITH CELL,
WHERE HE'S DONE SOME DUAL
RECOMBINATION MEDIATED LINEAGE
TRACING TARGETING ON THE SGTP1
AC1 AND CGP3 A2, SUBPOPULATIONS
IN CLUB CELLS AND INSURING THEIR
CONTRIBUTION TO THE LUNG
REGENERATION SO I THINK IT'S
POSSIBLE THAT IF WE HAVE MARKERS
--
>> I WAS THINKING OF SOMETHING
SIMILAR, THERE ARE NO REAL BASAL
CELLS IN THE DISTAL MOUSE AIRWAY
BUT THERE ARE SMALL POPULATIONS
OF P63 POSITIVE CELLS AND THOSE
CELLS CAN DIFFERENTIATE DOWN
DIFFERENT PATHS DEPEBDING ON
WHERE THEY FIND THEMSELVES SO
WONDERING IF YOU EVER TRIED TO
LOOK AT THE DUO BETWEEN SGBAP131
AND P63.
>> THANK YOU FOR THE QUESTION,
WE HAVE DONE RECENTLY A PAPER
SHOWING THAT CLUB CELLS WHEN
THEY MIGRATE OUT INTO THE
SURROUNDINGS ALVEOLAR REGIONS
THEY ARE UPREGULATION P6 A3, AND
THEY CONTRIBUTE TO THE AT2
CELLS.
YEAH, IT'S POSSIBLE BUT WE FIND
IN HOMEOSTASIS IN CONDITION, NO
PROXIMAL IN THE [INDISCERNIBLE]
REGION WE SET SUCH A P16
POSITIVE CLUB CELLS BUT ONLY
AFTER INJURY, WHEN THESE CLUB
CELLS INTEGRATE OUT, THEY
INCREASE THE LEVEL OF P6 A3.
YEAH.
>> SO MAYBE THAT ACCOUNTS FOR
THE CLUB CELLS POTENTIATING AND
DEVELOPING IN THE ALVEOLAR WHEN
THEY'RE ACTUALLY DEVELOPING A
REAM PHENOTYPE WITH P63.
>> YEAH.
>> DO HAVE YOU A QUESTION?
>> YEAH, JAY IS FINE.
>> RUBY IT'S NIGHT TO SEE THE
WORK COMING LIEWNG SO NICELY.
I HAD A QUESTION FOR YOU, IT'S A
GOOD THING JOHN IS HERE BECAUSE
YOU MENTIONED IONOCYTES
HETEROGENEITY AS 1 POSSIBILITY
BUZZ THERE IS 1 GROUP AS
ABNORMALITIES SOFGZ, 1 SAYING
SECRETION, 1 SAYING BI
CARBONATE, AND 1 THING I LIKED
ABOUT JOHN'S PAPER IS THAT IT
PERFORMED PROTURBATION AND
SHOWED THINGS CAN CHANGE.
SEE OIM WONDER FIGURE YOU THINK
A GIVEN IEE, AUDIENCEON O SIGHT
CAN SECRET AND ABSORB BOY MOVING
IT THROUGH.
BECAUSE THESE SAM INDUCED AND
STUFF LIKE, OR WHETHER THERE'S
LIKE FIXED HETEROGENERATED 8ITY
IN DIFFERENT POPULATIONS OF
IONOCYTES AND JOHN AFTER RUOBING
GIVES HER ANSWER, I WOULD LIKE
TO HEAR WHAT SHE HAS TO SAY.
>> THAT'S SUCH A GOOD QUESTION,
I JUST CAME BACK FROM IPSC, TO
WE'RE NOT SURE WHAT IT DOES BUT
IT DOES EVERYTHING, IT ABSORBS,
BIOCARBS, APPROXIMATE, H, SO I
THINK WOOER IN THE PROCESS OF
PERFORMING SINGLE CELL RNASEQ IN
OUR SYSTEM AND IT WILL BE NICE
TO SEE IF THE TYPES OF IONOCYTES
WERE ALSO IN THE PROCESS OF
GIVING OUR SYSTEM A PROTURBATION
IN THE DIFFERENCE IN
SALIVA--SALIVAINITY AND Ph AND
PROINFLAMMATORY PROTURBATIONS
AND SEE THE CHANGES.
SO THIS WORK IS ONGOING, YES,
AND SO JUST, PLEASE, YAWN, I
THINK YOU KNOW ABOUT THIS MORE,
MY UNDERSTANDING FROM THE TOP,
IS THAT THERE ARE THESE USE OF
DIFFERENT BASAL LATERAL -- I'M
NOT AN ELECTROPHYSIOLOGIST, I
WONDER IF WE DO OUR CHAMBER THE
SAME WAY THAT THE IOWA GROUP DID
IT THAT SHOWS THAT IONOCYTES
SECRETE, ABSORB, I WONDER WOULD
WE ALSO GET SIMILAR RESULTS?
AND I DON'T KNOW IF THAT'S A
GOOD ANSWER BUT I THINK IT'S --
I THINK THERE ARE SUBTYPES AND I
THINK THEY CHANGE WITH DIFFERENT
PROTURBATIONS, JOHN I WOULD LOVE
TO HEAR YOUR ANSWER.
>> A SUGGESTION AND MAYBE YOU'VE
DONE THIS ALREADY, BUT YOU'VE
ALREADY DONE A PROTURBATION
WHICH IS THE KNOCK OUT, AND THE
OTHER CELLS RESPOND TO THE LOSS
OF IONOCYTES.
>> YEAH, SO I THINK WE'RE IN THE
MIDDLE OF -- WE'RE GETTING
SINGLE SRNASEQ FROM OUR KNOCK
OUT CULTURES AND WILD-TYPE
KOWLTURES AND I'M EXCITED TO SEE
HOW ESPECIALLY THE SECRET ATORY
CELLS AND GIVEN TYPES OF CELLS
AND SOME OF THEM ARE HIGH AND
SO, YES, NOT JUST HOW SUBTYPES
OF THIS, BUT ALSO SUBTYPES OF
OTHER CELLS CHANGES WITH FOX AND
OUR SYSTEM DOES THAT ANSWER YOUR
QUESTION?
>> YEAH, BEFORE I COMMENT ON
JAY'S QUESTION, RUBY DO YOU KNOW
IN THE ELECTROPHYSIOLOGY THAT
YOU SHOWED, WAS IT APICAL LO
CHLORIDE OR SYMMETRICAL
CHLORIDE.
>> I THINK IT WAS SYMMETRICAL.
WE DID IT WITH YAY AND WE DID IT
WITH -- YEAH, I THINK IT WAS --
I'M PRETTY SURE IT WAS
SYMMETRICAL, AS SOON AS I SAW
THOSE DATA, I TALKED TO JAY AND
WE USED THE SYMMETRICAL FLUORIDE
GRADIENT.
>> RIGHT.
SO THE POSITIVE CURRENT YOU SEE
WOULD BE SECRETION OF CHLORIDE.
AND I'M SURE IF YOU VARY IN OUR
PAPER WE USE SYMMETRICAL
CHLORIDE SO THAT OUR OUR
IMPRESSION WE'RE PHYSIOLOGICALLY
RELEVANT, THERE ARE CHLORIDE
GRAYIENTS IN IN VIVO, BUT IF YOU
MANIPULATE IT AND YOU MACK THE
CHLORIDE, YOUR CURRENT WILL GO
UP AS YOU EXPECT, MAKE LOW BASAL
LATERAL CHLORIDE AND YOU SEE
FAIRLY LARGE NEGATIVE CURRENT,
TOO.
NYEAH, ABSOLUTELY, YEAH.
>> TO YOUR QUESTION, JAY, IT'S
REALLY INTERESTING.
I MEAN THERE ARE REGIONAL
ANATOMICAL DIFFERENCES TO THINK
ABOUT WHEN YOU THINK ABOUT
IONOCYTES.
THEY'RE ARK ROUND GLANDS SO
THEY'RE IN THE GLANDS IN THE
DUCTS AND WE NEVER KNEW THAT
WERE EVEN REALLY FAR DOWN IN
FREQUENT CELLS NEAR THE VERY
DISTAL END OF THE SNI, THAT THAT
EXPRESS FOX 1 AND LINEAGE TRACE.
SO THE REGIONALLITY DOES THAT
LEAD TO SUBTYPE OR IS IT A
STATE?
AND YOU KNOW THERE REALLY AREN'T
SUBTYPES BUT 3 DIFFERENT STATES
THAT ADJUST IS THE CELL LOCKED
INTO A TYPE OF PHYSIOLOGY.
WE DID THE HYPOTONIC AND
HYPERTONIC STRESSOT SITES AND IT
VERY MUCH CHANGES THE CHANNELS
THAT ARE EXPRESSED WITHIN THAT
THAT WOULD CONTROL SECRETION OR
ABSORPTION AND TO ANSWER THE
QUESTION, KNOCKING OUT FOXI 1,
AND CHANGES OTHER CELL TYPES AT
LEAST IN THE STUDIES WE DID, WE
FOUND THAT OTHER CELLS LIKE
SECRETORY CELLS WOULD ADOPT
HIGHER EXPRESSION OF CHANNELS
THAT WERE DIFFERENTIALLY
EXPRESSED AT HIGH LEVELS IN
IONOCYTES.
SO CERTAIN 3 THERE'S SOME
COMPENSATION BY THE EPITHELIUM
TO TRY ITS BEST EVEN IF IT CAN'T
COMPLETELY CORRECT TO MODULATE
OTHER CHANNEL ANDS GET BACK TO
HOMEIO STASES.
>> I'M NOT SURE HOW OR WHY 1
EXPECT IT TO BE THAT DIFFERENT
THAN THE KIDNEY.
THAT THEIR COMPENSATORY
MECHANISMS, IONOCYTES ARE
PLASTIC AND I WOULDN'T BE
SURPRISED IF THEY WEREN'T LIKE
ABLE TO CHANGE THEIR PHYSIOLOGY
AND RESPONSE TO AN ENVIRONMENTAL
CONSTRAIN.
>> YEAH, I TEND TO AGREE WITH
YOU BUT AT LEAST FOR OUR STUDIES
AND FOR DOING THAT IN FERRET
THERE ARE A NUMBER OF OTHER
DRIVERS WE HAVE TO MAKE AND I
WILL ADDRESS THAT.
>> WHILE WE WAIT FOR QUESTIONS,
WILL WE TALK ABOUT USING
MULTIPILL MODELS AND
TECHNOLOGIES, HOW DO WE TACKLE
MODEL SPECISKS THAT ARE MAY NOT
TRANSLATE INTO THE BIOLOGY.
WHERE DO WE START WITH THAT?
I WILL DRAW THE LINE OF WHAT
WE'VE SEEN AND WHAT STILL STANDS
OUT IN
>> THE SIMPLE ANSWER FOR OUR
ENGINEERED AIR WAY, I ALWAYS
VALIDATED WITH A PRIMARY AIR WAY
AND ALSO MODEL SYSTEM AND ONCE I
SEE THE PHENOTYPES IN A PRIMARY
ANIMAL SYSTEM, AT LEAST FOR THIS
VERY NEW PROTOCOL, THAT'S WHERE
I FEEL A LITTLE BIT MORE
CONFIDENCE AND MAYBE WHAT I'M
SEEING IS BIOLOGICAL BUT EVEN
THEN, I THINK IT'S RIGHT, I
THINK IT'S HARD TO INTERPRET AND
ESPECIALLY SORT OF GIVEN WHAT
QUESTION YOU ARE ASKING, I HOPE
THAT ANSWERS, TOO, TO SOME
DEGREE.
>> NO MODEL'S PERFECT.
AND MICE, OBVIOUSLY, YOU KNOW
YOU CAN GET, YOU CAN ADDRESS
DEEPER AND DEEPER MECHANISM THAN
YOU EVER COULD IN A FERRET.
AND WHEN THERE ARE DIFFERENCES
I'VE ALWAYS BEEN AN ADVOCATE OF
LEARNING THAT BIOLOGY BECAUSE I
THINK IT'S TELLING YOU SOMETHING
AND IT'S POTENTIALLY LIKE IN THE
CASE OF CYSTIC FIBROSIS,
PROVIDING OPPORTUNITIES TO
APPROACH THERAPIES FROM A NEW
ANGLE BECAUSE YOU UNDERSTAND WHY
MICE DON'T GET CYSTIC FIBROSIS
LUNG DISEASE WHEN YOU TAKE OUT
CFTR, YOU WILL NEVER GET AROUND
SOME OF THE ANATOMICAL CELL
BIOLOGY DIFFERENCES BUT EMBEDDED
THEY ARE REALLY INFORMATIVE
LESSONS, I THINK.
>> YEAH, I AGREE WITH JOHN.
TODAY I SEE THE FERRET MODELS
THAT HAVE LIKE TERMINAL
RESPIRATORY FROM DIFFERENT
MOUSE, I THINK MAYBE IN THE
FUTURE, WE CAN ALSO TRY TO
DEVELOP SOME MUSEUM MODELS ON
THIS NEW GENETIC MODELS ON THIS
ROAD WHICH IS REALLY PROMISING
AND MORE CLOSE TO HUMAN DISEASES
WHICH I LEARNED FROM TODAY'S
COMMUNICATION.
WE'RE MODELING OFF YOUR
EXCELLENT WORK.
>> I WANT TO MAKE 1 MORE COMMENT
ABOUT ORGANOID MODELS, I THINK
TO, ALSO GOING BACK TO 1 OF
JAY'S EARLIER COMMENTS ABOUT THE
STAGE OF THE ORGANOIDS, I THINK
RIDE NOW, THERE'S STILL MISSING
PIECES TO FURTHER RECAPITULATE,
THAT EARLY POSTNATAL PERIOD,
THERE'S A LACK OF OXYGEN
EXPOSURE, AND THERE'S ALSO A
LACK OF MECHANICAL STRETCH
BECAUSE AFTER BIRTH, RIGHT,
BABIES START TAKING THE FIRST
BREATH AND A LOT OF THINGS
ACTUALLY CHANGE ON THAT DAY IN
THE ALVERMEN INFECTED O LIAISON
THAT WILL KEEP DEVELOPING IN
RESPONSE TO THE
MICROENVIRONMENT.
THAT'S SOMETHING WE'RE TRYING TO
INCORPORATE INTO OUR CURRENT
SYSTEM, THE OXYGEN GRADIENT AND
USE A STRETCHABLE MEMBRANE TO
STUDY SOME OF THE MECHAN O
TRANSDUCTION IN THE ORGANOID
SYSTEM, POSSIBLY AND ALSO, YOU
KNOW THE STARTING OF THE
PULMONARY CIRCULATION, IS ALSO
SOMETHING WE THINK WOULD BE
IMPORTANT TO ADD INTO THE SYSTEM
TO BETTER RECAPITULATE IN HUMAN
PHYSIOLOGY.
>> I HAD A QUESTION ABOUT
ORGANOID, IT WAS REALLY
FASCINATING SINCE THE
VASCULATURE IS CAPABLE OF FLOW
THROUGH, DO YOU THINK YOU CAN
USE THAT TO STUDY IMMUNE
MOLLULATION, CAN YOU STUDY
IMMUNE INTERACTIONS HAVE YOU
TRIED THAT.
YEAH, ABSOLUTELY THAT'S AN AREA
WE'RE INTERESTED IN PURSUING,
FOR THE TIME BEING, WE'RE
BASICALLY TRYING TO OPTIMIZE THE
BARRIER FUNCTION OF THE
VASCULATURE WITHIN THE ORGANOIDS
AND AFTER WE, YOU KNOW DEVELOP A
BETTER JUNCTION, AND THAT'S
DEFINITELY SOMETHING WE CAN
FURTHER DEVELOP WHICH REQUIRES A
CONNECTION WITH A
[INDISCERNIBLE] DEVICE AND/OR
CURRENT THOUGHT IS TO YOU KNOW
CULTURE SOME MORE ROBUST VASC
DATURE ON THE CHIP AND THEN
CONNECT WITH THE VESSELS WITHIN
THE ORGANOIDS AND THAT WAY WE
HAVE AN INLET AND OUTLET AND
INTO THE SYSTEM AND HELP US
ADDRESS A LOT OF QUESTIONS.
AND MODEL A LOT OF SYSTEMS,
YEAH.
CONDITIONS.
>> I HAVE A QUESTION, ARE THESE
POSSIBLE?
[INDISCERNIBLE], YOUR SYSTEM IS
FASCINATING AND I WANTED TO ASK
A QUESTION REGARDING TO THE
MODEL THAT YOU ARE PROPOSING AT
THE END IN WHICH BY DISRUPTING
NOTCH SIGNALING WHAT YOU HAVE A
CONTRIBUTION THAT GOES
SPECIFICALLY TO THE TYPE 2 CELLS
OF THE BASCs, RIGHT, AND AS
OPPOSE TO THE TYPE 2 CELL, THE
CLUB CELL, GOING INTO ALVEOLAR
CELL ITSELF, THE TYPE 2 CELL, SO
THE ISSUE, THE NONAPOPTOTIC THE
ISSUE BUT THE OBSERVATION IS
THAT IN THE ADULT WHEN YOU
DISRUPT NOTCH SIGNALING, YOU
ACTUALLY DO HAVE A
TRANSFORMATION OF THE CLUB CELLS
BO THE CILIATED CELLS WHICH I
WOULD BE SURPRISED BECAUSE IN
SOME WAYS IT SHOULD ACTUALLY
HAPPEN FROM THE PROIENITTOR
ITSELF, NOT NECESSARILY FROM THE
ALREADY DIFFERENTIATED CELL,
UNLESS THERE IS SOME OTHER
MECHANISM I'M NOT SURE ABOUT,
BUT IF THAT'S THE CASE, THEN
WHAT YOU SEE IS THE
DISASSOCIATION BETWEEN THEZ
MECHANISM OF REGENERATION THAT
IS ORIGINALLY FROM THE TYPE 2
CELLS AND THE 1 THAT IS THE
CONTRIBUTION OF THE AIRWAY
ITSELF, GIVEN THAT YOU ARE NOT
SUPPOSED TO HAVE THEN CLUB
CELLS, TO GIVE RISE TO THE TYPE
2 CELLS, BUT THEN, IT IS THEN
THE BASC, THAT WE TAKE THE TYPE
2 CELL COMPONENT THAT IT HAS
THAT IT HAS THE SBC, SEON TO
REGENERATE THE ALVEOLAR TYPE 2
CELL.
I WONDER IF THIS IS CLEAR OR IF
I NEED TO EXPLAIN MORE?
YOU MEAN THE CLUB CELL
DIFFERENTIATING THROUGH THE BASC
CELL GOING THROUGH THE PHASE 2
STATE?
AM I UNDERSTANDING RIGHT IN.
>> GIVEN THAT YOU DO NOT HAVE
CLUB CELL OR YOU HAVE EITHER
DEFECTIVE CLUB CELL OR SO, IT
WOULD NOT BE ABLE TO OR GIVE
PRIZE TO A TYPE 2 CELL ITSELF
AND THAT IT IS WHAT IT IS
BLOCKED BY THE KNOCK OUT OF THE
NOTCH.
AND THE ONLY THING THAT WOULD
REMAIN WOULD BE THE BASC CELL
GIVING RISE THEN TO THE TYPE 2
CELL, YEAH, BECAUSE IT DOESN'T
HAVE THE COMPONENT OF THE CLUB
CELL THAT THE BASC WOULD HAVE
THAT IT IS [INDISCERNIBLE].
>> OKAY, YEAH, SO, IF WE KNOCK
OUT NOTCH, IN THE CLUB CELLS SO
THEIR CONTRIBUTION TO AT2 IS
SIGNIFICANTLY REDUCED SO IT'S
POSSIBLE THAT THE BASCs FOR NO
REASON, SENSE THIS DEFECT AND
THE COMPENSATED CONTRIBUTE MORE
TO AT2.
>> EXACTLY.
>> THERE'S DISASSOCIATION
BETWEEN THE 2 COMPONENTS, THE
COMPONENT THAT IT IS, WHAT YOU
HAVE IN MY VIEW, WHAT HAVE YOU
DESCRIBED VERY BEAUTIFULLY IS, A
COMPONENT THAT BELONGS TO THE
AIR WAY, AND IT ANOTHER 1 THAT
BELONGS TO THE BASC.
AND THAT CLEARLY, THE NOTCH IS
DOING THE SELECTION BETWEEN THE
2 COMPONENTS OF WHEN YOU HAVE
THE BLEOMYCIN REGENERATION,
REGENERATION AFTER THE
BLEOMYCIN.
>> YEAH, I THINK THIS TOO
POPULATION ARE FUNCTIONALLY
DIFFERENT IN THEIR RESPONSE TO
BLEOMYCIN INJURY BECAUSE IF THEY
RESPOND SIMILARLY, THEY SHOULD
BE DIFFERENT APPROXIMATE BASC
PART OF CLUB POPULATION,
SUBPOPULATION, THEY SHOULD
DECREASE THAT THEY ARE
DIFFERENTIATION INTO AT2 CELLS
WHEN NOTCH IS KNOCK OUT AFTER
BLEOMYCIN.
BUT WHAT WE SEE, BASICALLY IS
NOT CHANGE THEM, OR INSTEAD
INCREASE THEM, THE REASON FOR
INCREASE COULD BE AS A RESPONSE
TO REDUCE THE CLUB SALE
DIFFERENTIATION TO AT2, BUT IT
COULD ALSO BE TO THEY ARE
DIFFERENT POPULATION, THEY
FUNCTION DEFINITE BUT, WE THINK
THEY ARE NOT SAME POPULATION,
NOT ONLY FROM MOLECULAR
SIGNATURES BUT ALSO FROM
DISTINCT FUNCTIONAL DISTIRCHGHT
POPULATION FOR FUNCTION AS THEY
RESPOND DIFFERENTLY.
THIS IS WHAT WE CAN
[INDISCERNIBLE] FROM THIS DATA.
BUT AS YOU POINT OUT, I AGREE
WITH YOU, THAT IT'S ALSO
POSSIBLE THAT BASC INCREASE THE
BASC TO THE INTERNATIONAL
CLASSIFICATION IMRAIGZ TO AT2
CAN BE A SECONDARY TO REDUCED
[INDISCERNIBLE] CLUB AT2
REGENERATION.
THANKS OF.
>> SEUNG HYE HAS A QUESTION.
>> I HAVE A QUESTION FOR PANELS,
THAT IE. THINKING MORE LIKE
ALVEOLAR CELLS ACTUALLY BUT HOW
SIMILAR, THE INVITRO 3D SYSTEM,
CELL IS ACTUALLY -- HOW SIMILAR
TO THE INVITRO SETTING, SO IT
HAS VERY DIFFERENT METABOLIC
SETTING, COMPARED TO IN VIVO.
SO LIKE FOR EXAMPLE, IF WE LIKE
INDUCE DIFFERENTIATION, MY
LIMITED EXPERIENCE IS THAT THE
CELLS DOES NOT FULLY
DIFFERENTIATE INTO -- BECAUSE IT
LOOKED LIKE AT1 LIKE CELLS BUT
IT DOESN'T REALLY AT1 CELLS LIKE
IN VIVO, BUT IT CLEARLY, I HAVE
VERY LIMITED EXPERIENCE, SO I
JUST WANT TO ASK THE EXPERIENCE
IN PANELS, AND ALSO THE REASON
I'M ASKING IS THAT AT THE END, I
THINK IN TERPS OF THE RELATED TO
RESILIENCE IN OUR WORKSHOP, I
THINK AT THE END WE WANT TO KNOW
ONCE IT'S INITIATE THE
DIFFERENTIATION, WE WANT TO KNOW
WHAT ARE FACTORS TO MAKE THE
CELLS INVOLVING TERMINAL
DIFFERENTIATION, FULLY MATURE,
INTO FUNCTIONAL CELLS AND I
THINK THAT IF WE INVITRO SYSTEMS
CAN CAPTURE THAT PART, IT COULD
BE LIMITED IN DATA ASPECT, SO I
JUST WANTED TO ASK PANEL'S
EXPERIENCE OR OPINION ON THAT.
>> ALL RIGHT, I GUESS I CAN
START, FOR CURRENT ORGANOID, OF
COURSE DID BEPENDS HOW LONG WE
KEEP THEM IN CULTURE.
THE LONGEST WE TRIED IS ABOUT
ALMOST 4 MONTHS BUT THAT'S NOT
COMPLETELY INVITRO, WE TRAN
PLANTED THOSE ORGANOIDS AND THE
CAPSULE IN SKID MICE SO THEY CAN
FURTHER GROW AND CONNECT WITH
THE HOSTS CIRCULATION AND
MATURE, AND FOR THOSE ORGANIES
WE ARE DOING THEM TO UNDERSTAND
THE MATURATION OF THE CELLS ON
BUT IF THEY'RE TERMINALLILY
DIFFERENTIATIONS, IF THEY'RE
DIFFERENTIATED FROM IPSCs, 1
THING WE DID WAS TO COMPARE THE
TRANSI
TRAN SCRIPT OHM TO THE SYSTEM,
AND WE CAN TRANSFER IN VIVO FROM
GUESTATIONAL AGE A COUPLE WEEKS,
WEEK 4 OR 6, ALL THE WAY TO YOU
KNOW A THIRD TRIMESTER, HUMAN
FETAL LUNG, AND BASED ON
TRANSCRIPTOMIC ANALYSIS, IT'S
EQUIVALENT OR MORE SIMILAR TO
HUMAN FETAL LUNG AT ABOUT 80
DAYS OR SOMETHING LIKE THAT.
SO, OF COURSE, THIS IS STILL
VERY PREMATURE.
AND FOR USING THE MODEL TO
STUDY, YOU KNOW BIOLOGICAL
QUESTIONS, I THINK 1 POTENTIAL
ADVANTAGE OF THE SYSTEM,
ALTHOUGH IS NOW MATURE OR
TERMINAL DIFFERENTIATED, IT'S A
REDUCE DIMENSION SYSTEM, SO IT
AVOIDS COMPLEXITY OF THE IN VIVO
ENVIRONMENT, SO IF YOU WANT TO
UNDERSTAND SOME CELL AUTONOMOUS,
MECHANISM, IT MIGHT BE A
SIMPLER, PLATFORM, TO LOOK AT
FOR EXAMPLE, JUST ENDOTHELIAL,
EPITHELIAL CROSS TALK, SO YOU
DON'T HAVE, OTHER CELL TYPES,
INVOLVED IN THE SYSTEM, BECAUSE
WE KNOW, YOU KNOW ALL THE
PROGENITOR STEM CELLS IN THE
LUNG, THEY ARE PLASTIC AND THEY
CAN BE TRANSDIFFERENTIATED INTO
OTHER CELL TYPES WHICH INTRODUCE
A LOT OF COMPLEXITY, FOR
UNDERSTANDING A PARTICULAR
QUESTION.
SO IN MY VIEW, THESE IPS
DERIVATIVES ARE SERVING AS
SIMPLIFIED SYSTEM TO UNDERSTAND
AUTONOMOUS EFFECT.
>> YEAH, I HAVE TO SAY I
COMPLETELY AGREE WITH EVERYTHING
YOU JUST SAID I KNOW YOU'RE
QUESTIONS ARE FOR FOR THE
ALVEOLAR, BUT WHEN YOU COMPARE
AIRWAY TO UNCULTURED, FRESH
LUNG, AS WELL AS CULTURE TO
HUMAN BRONCHIAL EPITHELIAL
CELLS, THEY'RE SIMILAR WITH THAT
CORRECTION OF THE SINGLE CELL
DATA AND SO THEN ALSO WHEN WE
GIVE IT A PATHOGEN RESPONSE, WE
ALWAYS -- AT LEAST OUR LAB
ALWAYS COMPARED TO A PRIMARY
RESPONSE TO MAKE SURE IT'S NOT
AN IPSC PHENOMENON.
AND NOT FROM MY LAB BUT THE LAB
I TRAINED IN AT BU, TRANSPLANTED
IPSC DERIVED BASAL CELLS IN THE
MOUSE AND THEY WERE ABLE TO
DIFFERENTIATE INTO SOME OF THOSE
CELL TYPES IN VIVO.
SO YEAH.
I COMPLETELY AGREE WITH WHAT YOU
SAID MING XIA, ITIVE GOES A VERY
CLEAN WAY IF YOU COMPARE WITH
GENE EDITING IT'S A REDUCTIONIST
WAY TO ANSWER LIKE A ANSWER
SPECIFIC QUESTION.
>> YEAH, I GUESS OUR ALTERNATIVE
STRATEGY IS TO USE PRECISION CUT
LUNG SLICE, YOU CAN GET THOSE IN
CULTURE AND WITH AN ADULT LUNG
AS WELL AS AN INVITRO MODELING
SYSTEM.
>> I DON'T DENY THAT THIS SYSTM
IS HELPFUL TOOL TO ASK CERTAIN
QUESTIONS AND THEN UNDERSTAND
THE BIOLOGY, PATHOLOGY AND
POETICIENTIALLY USE IT AS A DRUG
SCREENING TOOL OR SOME
THERAPEUTIC TARGET BUT I CLEARLY
ALSO THERE ARE LIMITATIONS IN
TERMS OF HOW MUCH WE CAN MIMIC
IN VIVO SYSTEM, SO I JUST WANTED
TO DISCUSS ABOUT THAT.
I GUESS TATA HAS A COMMENT ON
THAT.
I ASKED HIM.
>> YEAH, IT'S NOT A QUESTION,
IT'S JUST A THOUGHT.
I WAS WONDERING WITH RESPECT TO
THE MATURITY, IS THERE SOMETHING
WE ARE MISSING, RIGHT IN 1
ASPECT IS YOU KNOW DURING
DEVELOPMENT AND DEVELOPMENT
PARTICULARLY, SO WE HAVE THIS
SELF-VERSUS NONSELF, WITH ALL OF
THIS IMMUNE TOLERANCE, RIGHT?
SO IS THAT COMPETENT BECAUSE I
REMEMBER SEEING SOME OF LAB
PAPERS WHERE I THINK THEY
POINTEDDA THE SOME OF THOSE LIKE
THEY DON'T EXPRESS, MHC PROFILE
IS DIFFERENT, ACTUALLY.
BUT IPSC DERIVED 2 AT2 CELLS,
BASAL CELLS THEY DON'T HAVE
THEEZ IMMHC PROFILES SO IF THAT
IS A WAY TO INDUCE AND IF THAT
IS ACTUALLY INDUCING SOME TYPE
OF MATURITY.
>> IME WONDERING ABOUT
COMPONENTS FROM THE IMMUNE
SYSTEM MOSTLY OR FOR A MATURE
SYSTEM?
>> YEAH, MHC COMPETENCE, RIGHT?
THAT'S HOW WE DEVELOP, DURING
DEVELOPMENT THIS SELF, VERSUS
NONSELF, I WONDER IF THAT AS A
ROLE IN MATURITY?
BECAUSE I REMEMBER LIKE EVEN A
TRANSPLANTATION STUDIES, FROM
DARYL'S GROUP, THE MORE
TRANSPLANTED, IBCs,
TRANSPLANTED INTO MICE, SHOWED
CHARACTERISTICS MORE CLOSE TO
THE ADULT.
NWELL, THANK YOU SO MUCH FOR
THIS, I THINK WE WILL CARRY THE
DISCUSSION OVER THE CHAT BECAUSE
WE HAVE TO GO TO THE SESSION 3,
BUT I THANK ALL THE PRESENTERS
IN A LIVELY DISCUSSION, THANK
YOU SO MUCH.
>> THANK YOU EVERYONE.
>> YEAH, WE WILL START SESSION
3.
THIS WILL BE ON PROGENITOR BASED
MARKERS PREDICTING EARLY CHRONIC
LUNG DISEASES.
IF WE GO TO THE NEXT SLIDE.
SO THIS SESSION WILL BE FOCUSED
ON LOOKING INTO PROGENITOR BASED
MARKERS THAT COULD PREDICT EARLY
ONSET OF CHRONIC DEC.
INITIALLY OF COURSE WE WILL HAVE
TO BE TALKING ABOUT RESEARCH
GAPS LATER ON BUT NEXT SLIDE,
PLEASE.
NEXT SLIDE.
NEXT.
SO THE FIRST PRESENTATION WILL
BE FOCUSED ON LOOKING AT AGAIN,
THE MARKERS OF PROIENITTOR BASED
MARKERS, THAT ACCOUNTED BE USED
FOR CHRONIC LUNG DEC, AND THE
SECOND PRESENTATIONS GOING TO BE
FOCUSED ON LOOKING INTO WHETHER
WE CAN USE BRONCHIAL RAVAGE
FLUID, PLASMA SAMPLES, THAT
WOULD PREDICT OR SHOW SOME
PATHOLOGICAL SOUTH STATES THAT
ARE FOULING THE LUNG AND WE WILL
HEAR ABOUT THESE 2
PRESENTATIONS, MAYBE YOU SHOULD
GO TO THE PRESENTATIONS
DIRECTLY.
THE FIRST PRESENTATION WILL BE
BY DR. MOUMITA GHOSH, ASSOCIATE
PROFESSOR OF MEDICINE AND IN THE
MEL MONEY SCIENCES AND CRITICAL
CARE UNIVERSITY OF COLORADO AND
SHE WILL TALK ABOUT DYSFUNCTION
OF AIRWAY BASAL PROGENITOR CELLS
IN YOUNGER SMOKERS WHETHER THAT
IS ASSOCIATED WITH AFFAIRS TEAM
LEADERRERRED BIOENERGETICS.
PLEASE GO,A HEAD.
>> THANK YOU.
THANK YOU FOR THE INTRODUCTION
AND I WOULD LIKE TO THANK THE
ORGANIZERS FOR THIS OPPORTUNITY.
SO I WILL TALK ABOUT DYSFUNCTION
AND, OPPORTUNITY, BASAL
PROGENITOR, AND ASSOCIATION WITH
BIOGENETICS AND I WILL ALSO LIKE
TO SHARE SOME NEW DATA TO SHOW
THAT HOW THIS COULD BE RELEVANT
FOR THE PATHOGENICITY OF EARLY
COPD.
SO AS WE ALL KNOW COPD,
CIGARETTE SMOKE IS THE MAJOR --
IDIO LOGIC FACTOR FOR COPD,
OTHER FACTORS CAN DEFINITELY
CONTRIBUTE TO THE DEC
PATHOGENESIS, IT INVOLVES 2
PATHWAYS TO THE LUNGS, AIRWAY
AND ALVEOLAR, COPD IS DONE BY
SPIRITUAL STRUGGLESOMETRIC
MEASUREMENT SO IT'S IMPORTANT TO
VEE WHERE THIS THE AREA IT IS,
SPIROMICS ROUGH ATOM METIC LUNG
FUNCTION DECLINE THAT DEFINES
COPD IS COMING FROM, SO ON THE
LEFT WE'RE LOOKING AT THE
DIFFERENT GENERATION OF THE
AIRWAY, SO FEV1 DECLINE WHICH WE
SEE IN THE COPD, THAT'S COMING
FROM 5 TO 6 GENE GENERATION OF
THE AIR WAY IN THE TERMINAL
BRONCHIAL AND THEN, THE
DIFFUSION ISSUE THAT HAPPENS IN
THE ALVEOLAR IN THE PARRINK MA
THAT LEADS TO LOSS OF DLCO, IN
SPITE OF [INDISCERNIBLE], YOU
ARE LOOKING AT [INDISCERNIBLE]
VOLUME AND IN THE COPD WE SEE
THAT THESE VERY CHARACTERISTICS
SCOOPED FEAT AND YOU ARE THAT
RELATES TO THE BASICALLY SAYS
THAT FIXED OBSTRUCTION THAT WE
SEE IN COPD AND WITH THAT IT'S
BASICALLY THIS RATIO OF FEV 1
AND IF WE SEE LESS THAN 70% THAT
IS THE DIAGNOSTIC FEATURE OF
COPD, I WILL JUST COMBACK TO
THIS, WHY THIS IS SO IMPORTANT
TO UNDERSTAND THAT WHERE IT'S
COMING FROM.
MOVING ON, NEXT SLIDE, PLEASE.
SO, WE HEARD A LOT ABOUT THE
BASAL PROGENITOR CELL, SO THIS
IS 1 OF THE MOST CRITICAL CELLS
IN THE AREA BECAUSE THEY ARE
INVOLVED BOTH IN THE MAINTENANCE
AND FOR THE REPAIR OF THE
ALVEOLAR AFTER AN INJURY, AND
THESE CELLS DO THEIR FUNCTION BY
2 WAYS, 1 THEY ARE HIGHLY
SELF-RENEWING, THEY CAN MAKE A
COPY OF THEMSELVES AND THEY'RE
ALL MULTIPOTENT SO THEY CAN GIVE
RISE TO ALL THE DIFFERENT CELL
TYPES PRESENT IN THE AIR WAY.
NEXT SLIDE, PLEASE.
SO A FEW YEARS AGO, WE STARTED
DOING A PROJECT WHERE OUR
HYPOTHESIS WAS TO SEE, BECAUSE,
THESE CELLS ARE SO IMPORTANT FOR
AIRWAY, REPAIR, THAT WHETHER
DYSFUNCTION OF THIS BASAL
PROGENITORS ARE PART OF THE
PATHOGENICITYS OF COPD, SO FOR
THIS STUDY, WE TOOK ADVANTAGE OF
BRONCO SCOPEY STUDY THAT GOING
ON IN COLORADO WHERE PEOPLE WHO
AREAT RISK FOR LUNG CANCER, ARE
UNDERGOING IN THE ENDOBRONCHIAL
BIOPSIES TO GET TISSUES FOR
BIOPSIES.
SO WE COLLECTED THEM AND LOOK
FOR PROGENITOR FUNCTION AND
THESE ARE THE 3 THINGS WE ARE
LOOKING FOR.
HOW MUCH PRESENTED IN THE BIOPSY
AND THE ABILITY TO SELF-RENEW
AND THEN LOOKEDDA THE ABILITY TO
DIFFERENTIATE TO ALL DIFFERENT
CELL TYPES AND ON THE TOP, YOU
ARE LOOKING AT THE TIME FRAME
THAT EACH OF THESE THINGS TAKES
AND IN THE CULTURE FROM THE TIME
OF DRONKIAL COPY EMPLOY SO NEXT
SLIDE, PLEASE.
SO WHEN IT'S COMPARED, SO THESE
PEOPLE, WHO HAVE SO NOW WE ARE
LOOKING AT THESE PEOPLE WHO HAVE
COPD, BI SPIROMETRY.
-- THEY ALL WERE SEVERELY
IMPAIRED IN COPD COMPARED TO
NONCOPD.
NEXT SLIDE, PLEASE.
NOT ONLY THAT, THE OTHER
INTERESTING THING THAT WE FOUND
THAT THE PROGENITOR COUNT WHICH
WAS THE VERY FIRST MEASURE WE
DID FOR PROGENITOR FOR HOW MANY
PROGENITOR PRESENT, WE FOUND
THAT PROGENITOR COUNT IS REALLY
HIGHLY ASSOCIATED WITH LUNG
FUNCTION DECLINE AND EACH WAY WE
COULD LOOK AT IF WE VIEW ON
RATIO, AND THE MOST INTERESTING
THING IS THAT THESE BIOPSIES
WERE TAKEN FROM [INDISCERNIBLE]
GENERATION OF THE AIRWAY,
WHEREAS THE LUNG FUNCTION
DECLINED WHAT WE ARE LOOKING AT
HERE, PARTICULARLY DIFFUSION,
THAT'S UPON HAING MUCH MORE
DISTAL TO THE LUNG SO WHAT WE
FOUND IS THAT LARGER PROGENITOR
RELATES TO THE PHYSIOLOGY THAT'S
HAPPENING MUCH DEEPER IN THE
LUNG.
NEXT SLIDE, PLEASE.
ONE MORE INTERESTING FEATURE
FROM THIS STUDY THAT WE FOUND
THAT WHILE ALL THE COPD, AIR WAY
BIOPSIES GAVE US VERY LITTLE
PROGENITORS, NONCOPD HAD A MUCH
HIGHER SPREAD, AND SO SOME OF
THEM, LIKE A GROUP, WE FOUND
THAT THEY ARE PROGENITOR COUNT
SIMILAR TO COPD, SO WE WANTED TO
KNOW, IF WE DIVIDE THIS GROUP
INTO NONCOPD HIGH PROGENITOR AND
NONCOPD LOW PROGENITOR AND HOW
THAT RELATES TO THEIR LUNG
FUNCTION AND HOW YOU SEE THE
RIGHT SIDE AND BASICALLY FOUND
THE PEOPLE WHO HAVE BIOPSIES
THAT HAD LOW PROGENITORS AND
THEY HAVE INTERMEDIATE LUNG
FUNCTION AND I'M SHOWING YOU HER
2 DIFFERENT FEATURES AND WE
LOOKED AT ALL DIFFERENT
MEASUREMENT OF LUNG PHYSIOLOGIC
AND EACH CARE WE FOUND THIS, LO
PROGENITOR BIOPSY, THEY HAVE --
THEY ARE COMING FROM THIS
INTERMEDIATE LUNG FUNCTION.
SO THAT GIVES US KIND OF AN IDEA
THAT THESE PEOPLE, MAYBE THAT
THEY ARE AT-RISK TO DEVELOP THE
DISEASE.
WE ALSO, THE NEXT THING WE DID
WAS WE WANTED TO UNDERSTAND WHAT
WAS THE MOLECULAR DIFFERENCE
BETWEEN THESE GROUPS OF
PROGENITOR CELLS.
NEXT SLIDE, PLEASE.
SO THE FIRST THING I WOULD LIKE
TO GET YOUR ATTENTIONOT LEFT
PART OF THE SLIDE, SO, HERE WE
ARE USING A BULK APPROACH TO
LOOK AT THE DIFFERENCE BETWEEN
NONCOPD HIGH AND PROGENITORS TO
SEE IF THERE ARE SOME KIND OF
LIKE A STABILIZED CHANGE IN THE
EXPRESSION OF DIFFERENT GENES IN
THESE 3 GROUPS AND WE FOUND THAT
THERE ARE 47 YEENS ONLY THAT ARE
SIGNIFICANTLY DIFFERENT BETWEEN
2 AND 3 GROUPS AND WHEN WE LOOK
AT THE PATHWAYS OR CELLULAR
FUNCTION THAT ARE IMPLICATED AND
REALLY WHAT CAN TURN OUT TO BE
MORE IMPORTANT IS THAT SIMILAR
GENERATEDET INCREASE IN BODYS
SEEM TO BE SOMETHING THAT'S KIND
OF CHANGING AS WE GO FROM LEFT
TO RIGHT.
SO PROMO AND INTEREST TO
ACTUALLY UNDERSTAND THE
DIFFERENCE IN BETWEEN NONCOPD
HIGH AND LOW PROGENITORS, WE
MIXED IT LIKE A TOY AN SIS
BETWEEN THIS GROUP WHICH IS IN
THE RIGHT AND NOW WE ARE SEEING
THAT ABOUT A HUNDRED AND 10
YEARS, WHICH ARE SIGNIFICANTLY
DIFFERENT BETWEEN THESE GROUPS
AND WHEN YOU LOOK AT THE
PATHWAYS, IT'S REALLY BECOME
OBVIOUS, THE DIFFERENCE BETWEEN
THESE 2 PROGENITORS SO THEY'RE
FUNCTIONALLY VERY DIFFERENT,
THEY ARE BIOGENETICS
PARTICULARLYOXIDATIVE
PHOSPHORYLATION, MITOCHONDRIA
MORPHOLOGY, SO EVERYTHING I
SHOWED YOU SO FAR, THEY'RE ALL
DONE IN A RELATEDLY OLDER
SMOKERS SO THEY ARE MEDIAN AGE
WAS ABOUT 60.
SO NEXT WE WANTED TO UNDERSTAND
WHETHER THE SAME MECHANISMS ARE
IN PLAY IN PEOPLE WITH
[INDISCERNIBLE] COPD.
NEXT SLIDE, PLEASE.
SO BEFORE I IMET TO THE DATA, I
WANTED TO KIND OF LIKE TALK
ABOUT WHO IS EARLY COPD, IT'S
REALLY A CONCEPT, THAT IT'S THE
CRITICAL TIME PERIOD WHEN
PATHOLOGIC MECHANISM ANDS THE
STRUCTURAL CHANGES ALL ARE
INITIATED AND THAT EVENTUALLY
DEVELOP INTO COPD.
SO THESE ARE REALLY CRITICAL
TIME, AND IF WE CAN UNDERSTAND
WHAT ARE THE PATHWAYS, MECHANISM
THAT ARE INVOLVED, NOW WE CAN
INTERCEPT THOSE, AND THEN
ACTUALLY DO SOMETHING OTHER THAN
AFTER THE DISEASE DIAGNOSIS,
WHEN ALL WE CAN DO IS TREAT THE
SYMPTOMS.
SO BECAUSE WE DO NOT HAVE A
DEFINITION, WE DO HAVE AN
OPERATIONAL DEFINITION, PUT
FORTH BY -- IN THIS PAPER, SO
THE VERY IMPORTANT THING IS
THESE ARE YOUNGER PEOPLE, SO
LESS THAN 50 YEARS OLD.
THEY HAVE 10 PACK CAREER SMOKING
HISTORY, AND THEY NEED TO HAVE 1
OF THE FOLLOWING FEATURES, POST
BRORCHGIAL DILATOR IS LESS THAN
OR LIMITED THAN NORMAL.
THAT'S REALLY LESS, WE DON'T
KNOW WHAT THE LOW LIMIT OF
NORMAL IS.
WE NEED TO HAVE RADIO LOGIC
ABILITIES AND REALLY TO FIND
EARLY COPD WE NEED TO DO A
LONGITUDINAL TO SEE IF THEY HAVE
RAPID DECLINE IN LUNG FUNCTION.
NOW TO GET THAT AND THE
KNOWLEDGE WE GAINED FROM THE
OLDER PEOPLE, OR SIMILAR THINGS
HAPPENING IN THE YOUNGER PEOPLE.
SO WE TOOK ADVANTAGE WITH THE
[INDISCERNIBLE] AND WE GOT CELLS
FROM OUR LOCAL [INDISCERNIBLE]
FROM BIOREPOSITORY WHERE THE
BIGGEST THING THAT TO LOOK AT
HERE THAT THE AGE GROUP OF THESE
PEOPLE ARE THEY'RE ALL BELOW 50
AND WE ONLY KNOW THAT THEY DID
NOT HAVE ANY KNOWN LUNG DISEASE.
SO FIRST WE GROW BASAL
PROGENITOR CLONED FROM THEM AND
THEN NEXT THING WE LOOKED AT IS
THE DIFFERENCE IN SELF-RENEWAL.
THE FIRST WHAT WE SAW THAT
NONSMOKER HAS MUCH HIGHER
SELF-RENEWAL, SIGNIFICANTLY
HIGHER SELF-RENEWAL, THAN
SMOKERS, BUT MORE IMPORTANT PART
WAS THAT WE FOUND JUST LIKE OUR
OLDER COHORT, THAT IN SMOKER HAS
A MUCH BIGGER SPREAD IN THEIR
SELF-RENEWAL.
SO WE WANTED TO DIVIDE THIS
GROUP INTO 2 GROUPS AND THAT
YOUNGER SMOKERS WITH LOW
SELF-RENEWAL AND WHEN YOU LOOK
AT THE DIFFERENTIATION IN THE
CULTURE WE FOUND THAT THE
DIFFERENTIATE THE FIND, THEY
MAKE A NORMAL MUCUS EPITHELIUM,
IN CONTRAST ISSUES THE LOW GROUP
MAKES A FLAT SCORE EPITHELIUM
AND THE EXPRESS [INDISCERNIBLE]
AND THEY BASICALLY FELL TO FULLY
DIFFERENTIATE BUT THEY DON'T
CRASH OR BURN, THEY JUST
BASICALLY AT A THERE.
THEY JUST FORGOT HOW TO
DIFFERENTIATE.
NEXT SLIDE, PLEASE.
SO TO GO BACK TO THE MECHANISM
WE SAW THE PROGENITORS SO WE
WANT TO LEARN THAT THERE IS A
BIG DIFFERENCE IN METABOLISM
FROM NONCOPD, SO WE DIVIDE THEM
INTO HIGHER, LOW, AND I GIVE
THOSE STUDIES PATIENT ID NUMBERS
HERE, SO JUST TO MAKE SURE, LIKE
SO TO MAKE IT CLEAR HOW
DIFFERENT THEY ARE, SO TO CREATE
A MORE RELATABLE SITUATION, WE
ALSO TREAT THESE CELLS WITH AIR
OR THE TRACT, THE MOST IMPORTANT
THING WE FOUND IN THE PRINCIPAL
ANALYSIS WAS WE FOWBD THAT THE
PROGENITORS BETWEEN
SELF-RENEWALS ARE BIOLOGICALLY
MORE DIFFERENT THAN CIGARETTE
SMOKE EXTRACT TREATMENT.
WE EXPECT IT DIFFERENT BUT IF
YOU LOOK AT THEY'RE ACTUALLY A
GROUPING BASED ON SELF-RENEWAL
RATHER THAN CIGARETTE SMOKE
EXTRACT TREATMENT.
AND WHEN YOU LOOK AT THESE IN
THE RIGHT MOST PANEL, WE'RE
LOOKING THE 15 MOST
SIGNIFICANTLY DIFFERENT
METABOLITES THAT ARE -- IF WE
LOOK AT, THIS WAS REALLY
SURPRISING THAT WE FOUND THAT
PROGENITOR WITH COMPOUND, ALL
HIGH METABOLITES, UTP, THEY ARE
EVERYTHING THAT GOES WITH HIGH
ENERGY THEY ARE PRESENT IN THE
LOW SELF-RENEW, RATHER THAN HIGH
SELF-RENEWAL GROUP EMPLOY SO
METABOLOMICS GIVE US THAT
[INDISCERNIBLE] WHAT IS
PRESENTING THE CELLS, SO WE
WANTED TO DO A MORE OF A KINETIC
ANALYSIS, NEXT, SLIDE, PLEASE.
SO WE USE THIS PROGENITOR CELLS
AND SEA HORSE STUDY, AND JUST AT
THE LEFT, YOU ARE LOOKING AT
JUST OVERALL, YOU KNOW
REPRESENTATIVE PICTURE OF HOW
THE RESPIRATION LOOKS LIKE AND
AS YOU CAN SEE THERE IS A
SIGNIFICANTLY HIGHER MAXIMAL
INSPIRATION AND THE LOW
SELF-GROUP COMPARED TO HIGH.
WHEN YOU LOOK AT EXTRA CELLULAR
[INDISCERNIBLE] HE FOUND THE
SAME THING SO THESE CELLS ARE
FIRING IN THE ALL DIRECTION TO
MAKE MORE AND MORE ENERGY AND WE
ALSO FOUND THAT SELF-RENEWAL IS
NEGATIVELY CORRELATED WITH BOTH
OXYGEN COMSUMPTION OR FOR DPLIK
O LYTIC FLUX.
SO THE OTHER THING IS THE
MORPHOLOGY, SO WE USE DIFFERENT
KINDINGS BOGHT ARE SIGNIFICANTLY
HIGHER IN LOW SELF-RENEWING
GROUP COMPARED TO HIGH
SELF-RENEWING GROUP.
NEXT SLIDE, PLEASE.
WE ALSO DID ELECTRON MICROSCOPY
TO BED THAT LIKE IN A REALLY
GETTING TO THE STRUCTURE OF
MITOCHONDRIA, AND WE FOUND THAT
LOW SELF-RENEWAL -- RENEWAL
PROGENITORS HAS THIS ELONGATED
FEATURE WHICH GOES WITH THEIR
FUSION, SO BASICALLY THESE MIOF
THE ARE FUSING WITH EACH OTHER
AND MAKING THIS LONG ELONGATED
STRUCTURE.
NEXT SLIDE, PLEASE.
SO BASICALLY TO SUMMARIZE WHAT
WE FOUND IS THAT PROGENITOR
DISCUSSION HAPPENS IN A SUBSET
OF YOUNGER SMOKERS AND
SURPRISINGLY, THIS LOW
SELF-RENEWING PROGENITORS
REQUIRE MORE ENERGY AND IT LEAS
TO ELONGATED STRUCTURE WHICH IS
PROBABLY CLEARS FOR THE ENERGY
REQUIREMENT.
NEXT SLIDE.
SO THE GAP HERE IS TO TRY TO
UNDERSTAND WHAT ARE THE
MECHANISM THAT DRIVING THIS?
AND MORE IMPORTANTLY HOW ARE
THAT I RELATED TO COPD
PATHOGENESIS.
THAT LEADS TO OUR ONGOING STUDY
WHERE OUR TARGETED GROUP IS
30-50 YEARS AND WHERE WE ARE
ENROLLING SMOKERS WITHOUT ANY
KNOWN LUNG DISEASE AND THEN
HAVING CONTROL NEVER SMOKERS AND
COPD SMOKERS, WE ARE COLLECTING
THEIR ENDOBRONCHIAL BIOPSY AND
NAISAL BRUSHINGS TO MAKE IT MORE
REAL LIFE SITUATION SO WE DON'T
HAVE TO BRONCH EVERYONE AND THIS
YEAR WE ARE LOOKING AT
PROGENITOR AND MEASURES FOR
COUNT, SELF-RENEWAL
IVRENTIATION, THIS WE ARE ALSO
THE PEOPLE ARE GOING THROUGH A
VERY COMPREHENSIVE OF CLINICAL
VISION, LUNG FUNCTION, AND THEN
THESE PEOPLE ARE GOING TO COME
BACK IN EACH YEAR, AND THEN
WHERE WE ARE GOING TO DO THEIR
BRONCHIAL, PULMONARY FUNCTION
AND ALSO COLLECT NAISAL
BRUSHINGS, AND WE WILL ONLY DO
BRONCHOSCOPY IF THERE IS AN
UNEXPECTED DECLINE OF LUNG
FUNCTION.
SO THIS STUDY IS ALREADY
RECRUITING PATIENTS AND THEN
HOPEFULLY WE WILL UNDERSTAND THE
IMPORTANCE OF THE BIOLOGY THAT
WE LEARN FROM OLDER COHORT, HOW
THAT IS RELATED TO EARLY COPD.
NEXT SLIDE, PLEASE.
AND I WOULD JUST RIKE -- LIKE O
THANK THE PEOPLE WHO PARTICIPATE
INDEED THE WORK, MY COLLABORATOR
ANDS FUNDING.
THANK YOU IF ARE YOUR ATTENTION.
THANK YOU MOUMITA FOR A
WONDERFUL TALK.
SO LARGE AIRWAYS BEING AFFECTED
SOMEHOW BY WHAT'S HAPPENING AT
THE LOWER AIRWAY, WITH
SIGNATURESIET SMOKE, THE LOWER
AIRWAY DISCIPLINARY STROID
SOMEHOW EMPLOY SO THANK YOU.
THE NEXT TALK WILL BE BY
DR. HERBERT SCHILLER, DIRECTOR
PRECISION REGENERATIVE MEDICINE
IN MUNICH AS WELL, HERBERT GO
ASHES HEAD.
>> THANK YOU.
FIRST OF ALL THANKS A LOT FOR
INVITATION THIS HAS BEEN REALLY
-- IT'S REGULATED.
SO RESILIENCE COULD BE DEFINED
AS THE ABILITY TO GO BACK AND
FORTH BETWEEN THESE INJURYITATES
HERE AND THE HEALTHY AND MY LAB
TRIES TO USE SINGLE SEARCH
ECONOMICS TO DISSECT THE
CELLULAR CIRCUITS THAT DEFINE
SIGNALS PATHWAYS AND MECHANISMS
THAT DEFINE THESE BARRIERS
BETWEEN DIFFERENT TISSUE SPACE,
SO HOW CAN WE GET LOCKED INTO AN
IRREVERSIBLE CONFIGURURATION HOW
CAN WE REPROGRAM THAT AND ALSO
FOR PURPOSES OF THIS TALK, I
WANT TO STRESS IT'S IMPORTANT IF
YOU COULD GO NEXT, PLEASE, THAT
IT'S VERY IMPORTANT CLINICALLY
TO READ OUT THESE TISSUE STATES
IDEALLY PERIPHERY FROM THE
BLOOD.
NEXT.
SO HOW CAN WE UNDERSTAND HOW
DIFFERENT TISSUE STATES ARE
REPRESENTED IN THIS SECRETED
PROTEINS FOR INSTANCE, IN
GENERAL MY LAB TRIES TO SCALE
EXPERIMENTAL PROTURBATIONS
MOSTLY IN PRESUGZ CUT LUNG
SLICES TO SIGNIFY SECTIONAL
ANALYSIS MECHANISMS USING SINGLE
SEGENOME MODEL CITIZENNICS TO
READ OUT THE TRANSCRIPTIONAL
STATES BUT WE CAN ALSO CORRELATE
SUPERINATENT [INDISCERNIBLE] PCS
AND SECRET OMICS.
SO WHAT WE STARTED TO DO WAS
TAKE DIFFERENT TISSUES CAPTURE
THEM EXVIVO AND DO ALL TYPE OF
GENETIC AND PUSH TURB EGGS AND
AS I SAID WE COULD LEARN
SOMETHING ABOUT THE BIOMARKERS
AND DIVISION WE HOPE TO USE THE
MODELS SO THAT THE NEXT PHASE OF
HUMAN CELL ATLAS IS PROBABLY
BASED ON ALL THESE DATA SETS
THAT CAN BE USED TO TRAIN
FOUNDATIONAL MODELS AND LEARN
ABOUT CAUSAL CELL AND TISSUE
BIOLOGY SO WE WANT TO PREDICT
DIFFERENT COMBINATIONS OF
PROTURBATIONS THAT CAN INDUCE
CERTAIN TISSUE STATES.
NEXT.
AND OF COURSE THIS IS AN ICS
VIVO MODEL, EXPERIMENTAL MODEL
SO WE LOOK AT THE PATIENT AND WE
TRY TO UNDERSTAND INSITU IN
PATIENT TISSUES WHAT WHAT
HAPPENS IN THE EARLY STAGE OF
THE DISEASE AND EVENTUALLY WE
NEED TO DESIGN BETTER STUDIES
WHERE WE CAN MAYBE FOLLOW
LONGITUDINALLY AND FOLLOW
THERAPEUTIC TREMES TO SEE HOW
THESE CELLULAR SECOND QUARTER
UTRS IN THE LUNG ARE RESPONDING.
NEXT.
SO 1 OF THE APPROACHES WE USE IN
COLLABORATION WITH THE
[INDISCERNIBLE] IN BELGIUM AND
[INDISCERNIBLE] DID A POST DOC
IN MY LAB WHERE THEY ARE REAL
SPECIALISTS IN DOING THE
MICROSTAGING ON IPS LUNG PLANTS
SO YOU CAN SEE THESE CORES THAT
WERE DRILLED FROM THESE FROZEN
LUNGS THAT WERE AIR INFLATED
OVER LIQUID NITROGEN AND THEN
THE MICRO CT SCAN CAN GOVERN A
VERY PRECISE QUANTIFICATION OF
THE TISSUE MODELING SO NEXT, SO
IT TOOK THE LUNGS AND 6 COURSE
PER LUNGS AND CONTROL LUNGS WITH
SEVERAL COURSE PER LUNG, SO THAT
WE ANALYZE A TOTAL OF 164 OF
THESE CORES.
NEXT.
AND SO WE DO THIS PROGRESSION,
SO WHAT I WANT TO EMPHASIZE HERE
IS YOU CAN APPRECIATE IN THE
GRAPHOT LEFT, THE SURFACE
DENSITY VERSUS THE TISSUE
PERCENTAGE, SO, BASICALLY THE
ALVEOLAR SURFACE AREA IS GOING
DOWN WITH DISEASE PROGRESSION
WHILE THE PERCENTAGE OF TISSUE
MASS IS GOING UP AND YOU SEE
THIS GRADUAL CONTINUATION OF THE
TISSUE REMODELING THAT -- AND
ALL THESE SAMPLES WERE SEQUENCED
BY THE NUCLEI R, INA SEQUENCING
EMPLOY NEXT.
OKAY SO THAT'S ALL THE CELL
TYPES, NEXT.
AND AS AN EXAMPLE WE CAN GO INTO
THE EPITHELIAL CELL TYPES, YOU
CAN SEE THE AT2 CELLS IN ORANGE
AND 1, AND THE 1S IN BLUE HERE
AND NEXT.
WHAT WE CAN DO IS WE CAN
SUBCLUSTER INTO DIFFERENT
TRANSCRIPTIONAL SUBSTATES OF
THESE CELL TYPE IDENTITIES AND
IF WE GO TO THE NEXT SLIDE THESE
RED AND BLUE COLORS INDICATEOT
LEFT SIDE, YOU CAN ALSO CLICK
ONCE MORE, ON THE LEFT SIDE,
WHERE THE CERTAIN NEIGHBORHOOD
OF CELLS IS ENRICHED IN DISEASE
OR IN THE HEALTHY CONTROLS AND
ON THE RIGHT SIDE, YOU CAN SEE
WHETHER IT'S THE CELLULAR
NEIGHBORHOOD IS ENRICHED IN
ENSTAGE DISEASE VERSUS EARLY
STAGE DISEASE AND YOU IT'S NICE
TO SEE THAT SINCE HERE IN THE 82
CELLS WE CAN IDENTIFY SUBSTATES
OF 82S THAT ARE VERY MUCH
ENRICHED IN VERY EARLY DISEASE
OR IN THE GRAY COLOR THOSE WOULD
ALL FIND IRRESPECTIVE OF THE
TISSUE DEGREE MODELS.
BUT THEY'RE QUITE DISTINCT FROM
NORMAL HEALTHY 82 CELLS.
NEXT.
SO WHAT WE DO NOW FOR ALL THE
DIFFERENT LINEAGES, WE THINK
THESE SUBCLASSES, SUBIDENTITY
CELL STATE ANDS CORRELATE THEM
WITH THIS CONTINUOUS DISEASE
PROGRESSION, FROM THE MICROCT
STAGING.
[MULTIPLE VOICES SPEAKING ]
>> NEXT, IF YOU COULD CLICK
IMEAN.
UNFORTUNATELY LOTS OF
ANIMATIONS.
NEXT.
SO ON THE X-AXIS, THE HEALTHY
DEC, AND THE Y-AXIS DISEASE
SEVERITY.
SO IN THE LOWER LEFT RECK TANGLE
YOU HAVE CELL STATES THAT OCCUR
IN OUR HEALTHY AND THE
EPITHELIAL STATES ARE IN BLUE,
AND YOU CAN CLICK 3 TIMES MORE.
SO THEN OF COURSE, WE CAN SEE
THE CELLS CO OCCUR AND VERY
SEVERE DISEASE, OR IN VERY MILD
OR EARLY DISEASE AND THEN ALSO
SOME CELLITATES ARE PRESENT
THROUGHOUT INCLUDING THE
ABERRANT BASAL CELLS WE HAVE
ALREADY HEARD ABOUT TODAY.
NEXT.
SO NOW WE CAN TRY TO INFER
DIFFERENT GENE REGULATORY
NETWORKS COMPUTATIONALLY AS
CELLS HAVE COMMONICATION NEOF
THE WORKS AND CHANGES PROGRAMS
AND WE ALSO WANT TO MAP THESE
CELL STATES INTO SPACE, SO THAT
WE CAN SEE, WHETHER THESE CELL
SETS NOT ONLY TALK HER IN AND
CALLED THAT DEC PROGRESSION TIME
BUT ALSO IN THE PATIENT
[INDISCERNIBLE].
NEXT.
SO FOR THE REST OF MY TALK I
WANT TO DISCUSS HOW MAYBE THESE
DIFFERENT CELL STATES ARE
REPRESENTED IN THE PERIPHERAL
BODY FLUIDS.
OF COURSE THAT'S AN IMPORTANT
QUESTION.
GO TO THE NEXT SLIDE.
BECAUSE IT WOULD BE CLINICALLY
VERY USEFUL AND IN THIS PAPER,
ALSO DUE TO THE TOPIC, I TAKE
THE OPPORTUNITY TO HIGHLIGHT
THIS PAPER, AGAIN WHICH WE
PUBLISHED A FEW YEARINGS AGO,
WHERE WE MADE USE OF MASS SPEC
QUANTIFICATION OF BRANCHING O
SAL VERMEN INFECTED OULAR
LAVAGE, AND WE INTEGRATED OUR
OWN SINGLE CELL TRANSCRIPTOMIC
DATA OF IPS PATIENTS WITH 2
OTHER DIFFERENT INDEPENDENT
COHORTS TO SORT OF LEARN THE
REPRESENTATION OF THE DIFFERENT
CELL STATES IN THE PERIPHERAL
FLUIDS AND SHOW YOU QUITE WELL
WORKED FOR SOME EXAMPLES.
NEXT.
SO FOR THE BRONCHIAL LAVAGE
FLUID, THEY TOOK THE FLUID FROM
THE PROTEINS AND THE DEALS
AROUND A THOUSAND PROTEINS AND
THIS WAS A 128 AND PATIENTS ON
THE INITIAL DIAGNOSIS, SO
REPRESENTING RATHER EARLY STAGES
OF THE DISEASE.
NEXT.
IN SHOWS DECONVOLUNTEERSULATION
OF THESE PROTEINS WITH SINGLE
CELL TRANSCRIPTOMIC MARKER
SIGNATURES.
YOU CAN SEE THIS MAKES SENSE IF
WE HAD SOME [INDISCERNIBLE]
CONTROLS HERE AND ALL THESE
DIFFERENT STROMAL CELL
POPULATIONS INCLUDING THE MY O
FIBROBLASTS, APPARENTLY GIVE
RISE TO SOME PROTEINS IN THE
ILD, ALSO SAME TRUE FOR THE
BASALOID CELLS.
NEXT.
SO HERE'S EXAMPLES FOR
CORRELATION OF [INDISCERNIBLE]
SO THEOX GENERATEDDATION
CAPACITY WITH ALL THESE
DIFFERENT MARKERS HERE WE
MEASURE IN THE LAVAGE FLUID AND
YOU CAN SEE ON THE AVERAGE MORE
NEGATIVELY CORRELATED WITH THE
PLCO AS THE OTHER ENRICHED
PROTEIN THAT WE MEASURED, RANDOM
PROTEIN WE MEASURED.
NEXT, THIS IS STILL JUST THE
LUNG, SO WE HAVE OTHER LUNG
FUNCTION PARAMETER SO WE CREATED
A METALUNG ORGANOID FUNCTION AND
WHERE WE AMERICANED THESE AND
CORRECTED FOR [INDISCERNIBLE]
CHAMBER.
NEXT, ALL THESE PATIENTS ARE NOW
ON A DIFFERENT MAYBE POINTING TO
THE TRAJECTORY OF LOOSING THE
LUNG FUNCTION DUE TO DISEASE.
NEXT.
AND SO THAT IT SHOWS THE
ENRICHMENT OF OR THE POSITIVE OR
NEGATIVE SORT OF CORRELATION OF
THE PROTEINS MEASURED IN THE
LAVAGE GROUP, WITH LUNG
FUNCTION.
SO THE MORE PROTEINOT RIGHT, THE
BETTER THE LUNG FUNCTION IN THE
ILD PATIENTS AND ON THE RIGHT,
WHICH ARE KNOWN PROTEINS
UPREGULATED IN IPS, NEXT.
NEXT.
AND WE HAVE MANY MORE CLINICAL
FEATURES AND 72 PROTEINS HAD A
GOOD CORRELATION WITH AT LEAST 1
OF THESE CLINICAL FEATURES, SO
HERE YOU CAN APPRECIATE THE
[INDISCERNIBLE] 1 IS
HIGHLIGHTED, IT'S VERY
NEGATIVELY, VERY POSITIVELY
CORRELATED WITH LUNG FUNCTION
BUT ALSO OTHER THINGS LIKE FOR
EXAMPLE, THE CD4, CT8 RATIO IS
VERY PREDICTIVE OF ALL THESE
PROTEINS THAT CORRELATED WITH
LUNG FUNCTION, NEXT.
SO THE QUESTION WAS IF WE USE
THE PROTEIN FEATURES WILL IT
PREDICT CLINICAL FEATURES ALSO
IN TRANSCRIPTOMIC DATA IN OTHER
COHORTS WOE WE TRAINED A RANDOM
MODEL AND APPLIED THAT MODEL TO
OTHER DATA SETS.
NEXT.
INCLUDING OUR OWN, SINGLE CELL
DATA AND A CONTROL FROM CHICAGO,
WHERE CORRECTLY THE REDUCED LUNG
FUNCTION WAS PREDICTED IN THE
SINGLE CELL TRANSCREPT ORDER OF
MICRONSIC DATA BUT THE TRAINING
WAS IN THE PROTEOMIC DATA SO
THERE WAS AN INFORMATION
TRANSFER THAT WORKED.
NEXT.
AND MUCH MORE IMPORTANTLY, THIS
ALSO WORKED IN THESE MICROCT
STAGINGS WHERE IT'S SUITABLE
PROGRESSION BECAUSE IT'S WITHIN
PATIENT, BUT IT CORRECTLY
PREDICTS A GRADUAL DECLINE IN
LUNG FUNCTION DEPENDENT ON THE
DEGREE OF TISSUE MODELING
INDICATING THAT INDEED THESE
BIOMARKERS REALLY REFLECT AND
REPORT TISSUE REMODELING.
NEXT.
SO WHERE'S THE SIGNAL COMING
FROM.
CAN YOU DO CORRELATIONS TO
PROTEOMIC DATA AND THE SINGLE
CELL TRANSCRIPTOMIC DATA AND
WOULD SUGGEST THAT THE BASAL
CELLS AND HAD EVEN MORE
CONTRIBUTION TO THE SIGNAL, AND
FOR EXAMPLE, THE FIBROBLASTS
WHERE I SHOWED IN EXAMPLES
BEFORE.
NEXT.
SO FOR THE ALVEOLAR, WE HAVE
HEARD ABOUT THE BASAL CELLS AND
THE WE THINK THE CELLS CAN GIVE
RISE OF THESE CELLS AND THE
MARKERS INCLUDING THE PROTEIN,
WOULD COME UP, WITHIN THAT
TRAJECTORY.
NEXT.
AND THAT CAN ALSO BE SEEN WHEN
YOU LOOK AT THE HETEROGENEITY
AND IPF, THAT THIS
ANTICORRELATES WITH THE
[INDISCERNIBLE] PROGRAM.
SO WE GET MORE INTERESTED IN THE
CRTAC PROTEIN, WHICH POSITIVELY
CORRELATED WITH THE LUNG
FUNCTION.
NEXT.
SO IT'S EXPRESSED MOST HAD THE
LUNG, IT'S EXPRESSED QUITE
SPECIFICALLY IN AT2 CELLS, DOWN
REGULATED IN ALL 3 COHORTS IN
THE AT2 CELLS AND VERY
IMPORTANTLY IT'S ONLY DOWN
REGULATED IN THE ILD, AND NOT IN
COPD, NEXT.
AND IF YOU CORRELATE WITHIN ALL
THIS C-POSITIVE 82 CELLS, THE
CPF PROTEIN WITH ALL YEENS AND
ON THE X-AXIS YOU SEE THE
CHICAGO COHORT AND ON THE Y-AXIS
YOU SEE THE NATIONAL COHORT AND
THEY'RE COLOR COATED AND THEY
ALL AGREE ON THESE GENES HERE,
SO THE [INDISCERNIBLE] IS A
PROGRAM AND ANTICORRELATE WIDE
GENES SUCH AS SOX 4 AND NEXT AND
NEXT, AND HERE CAN YOU SEE
DIFFERENT PATHWAYS THAT ARE
POSITIVELY OR NETIONATIVELY
CORRELATED WITH THE
[INDISCERNIBLE] IN 82, AND FOX
FORWAS INTERESTING BECAUSE IT'S
THE MOST NEGATIVELY REGULATED
ANTICORRELATED TRANSCRIPTIONAL
REGULATIONULATOR, NEXT WHICH WE
KNEW FROM THE MOUSE STUDIES SO
WHEN YOU MODEL, THE ACTIVATION
OF AT2 CELLS IN THE BLEOMYCIN
INJURY MODEL AND GOING TO THESE
TRANSITIONAL CELLS THAT WE HEARD
A LOT ABOUT TODAY, YOU CAN SEE
SOX 4, AND IT WOULD COME UP IN
THIS TRANSITIONAL CELLS.
AND NEXT, THE VERY SAME GENES
COME UP IN THE HUMAN PHASE AND I
WOULD SUGGEST THESE CELLS, AT
LEAST FUNCTIONAL OR TO LOOK TO
THE IDI CELLS IN YOU HUMAN CASE
AND IT SEEMS THE 1 LOSS WITHIN
[INDISCERNIBLE] SO VERY EARLY
MARK OFF OF THE TRANSITION.
NEXT.
SO CAN THIS BE SEEN IN THE
BLOOD?
WE MEASURED 2 PLASMA COHORTS,
NEXT AND IN THIS PLOT YOU CAN
SEE HOW PROTEINS DATA DETECTED
BOTH IN THE LAVAGE AND IN THE
PLASMA, AND THEY ARE STRONGLY
MORE EXPRESSED IN THE PLASMA OR
LAVAGE, [INDISCERNIBLE], THEY
ARE LOCALLY PRODUCE AND NOT JUST
[INDISCERNIBLE] FROM THE LUNG
AND THIS INCLUDES SURFACTIN
PROTEIN B AND [INDISCERNIBLE]
FOR THE CELLS AND MANY MARKERS
WE KNOW INCLUDING THE CRTAC 1
PROTEIN.
NEXT.
AND THEN ALSO IN THESE 2,
INDEPENDENT COHORTS THAT WERE
ACTUALLY QUITE DIFFERENT, 1
SHOWED THAT POSITIVE CORRELATION
WITH LUNG FUNCTION PARAMETERS IN
BOTH IN BOTH OF THESE COHORTS.
NEXT.
SO WHAT I'VE SHOWN YOU IS THAT
WE CAN FIND GOOD BIOMARKERS THAT
SEEM TO REFLECT THEITATE WHICH
CRTAC 1 IS EXAMPLE FOR LUNG
PROYINITTOR BASED BIOMARKER THAT
REPORPTS LUNG FUNCTION DECLINE
IN ILD SPECIFICALLY.
UF COURSE WE NEED BETTER
DESIGNED LONG NUDEINAL SETTING
TO STUDY THESE THINGS MUCH MORE
AND ALSO LARGER STUDIES TO
UNDERSTAND HOW DIFFERENT
CO-MORBIDITIES WILL AFFECT THESE
MEASUREMENTS, AND I WOULD
SUGGEST THAT MAYBE ALSO THIS PC
LS PERTURBATION STUDIES WILL
HELP TO PREDICT BIOMARKERS
PANELS TO MAKE SENSE TO LOOK FOR
IN THE BLOOD.
SO WITH THIS I'M DONE AND THE
NEXT SLIDE, I HAVE A FEW
ACKNOWLEDGMENTS OF THE TEAM AND
I THANK YOU FOR YOUR ATTENTION
AND HAPPY TO TAKE ANY QUESTIONS.
>> THANK YOU HERBERT, GREAT
TALK.
NOW WE CAN OPEN THIS FOR
DISCUSSION I WAS VERY INTERESTED
IN ACTUALLY ASKING YOU HERBERT
SPECIFICALLY NOW, YOU KNOW IN
THE FIRST PART OF YOUR TALK,
ALTHOUGH THE SIVENGLE CELL RNA
SEQ YOU ANALYZED YOU FOCUSED ON
THE TYPE 2 CELLS.
AND I WONDERED IF YOU WOULD HAE
LOOKED AT THE LARGE AIRWAY
EPITHELIAL CELLS IN THESE
DISEASED INDIVIDUALS AS IT GETS
MORE SEVERE OR MORE DECLINE,
REGARDLESS OF HOW YOU LOOK AT
IT, WOULD YOU BE SEEING SOME
KIND OF A LET'S SAY, SEVERE OR
SEVERITY ALSO IN THE LARGE
AIRWAY EPITHELIAL, SEMESTERULAR
TO WHAT, YOU KNOW WHAT MOUMITA
WAS TELLING US?
>> YEAH, THANKS FOR THE
QUESTION.
SO HERE, AT2S WERE GIVEN IN
DIFFERENTITATES OF COURSE I
THINK THEY'RE VERY IMPORTANT FOR
IPF.
BUT OF COURSE WE SEE LOTS OF
DIFFERENCES IN THE CELL TYPES,
THIS COMPLICATION THAT DUE TO
THE BRONCHIALIZATION, PROCESS,
YOU SEE WITH DISEASE
PROGRESSION, YOU SEE ALL THESE
AIRWAY PHENOTYPES AM CANNING UP
BUT YOU KNOW THE PAPER WOULD
SUGGIEST THIS COULD ALSO BE
DERIVE FRIDAY 82 CELLS, BUT
YEAH, I CERTAINLY THINK THE
AIRWAY ALSO CHANGES AND ACTUALLY
WHEN WE LOOK AT -- SO WE DO
LASER CAPTURING OF THE FOCI FOR
INSTANCE AND WE CAN SELECT SOME
FROM THE ALVEOLAR STAGE AND
EARLY STAGE COURSE, OR FROM THE
AIRWAYS AND THEY'RE A BIT
DIFFERENT PROFILES AND SOMETIMES
WE FIND INTERESTING THINGS LIKE
ON THE AIRWAY SIDE, YOU HAVE THE
BASAL CELL LAYER AND A BASAL
LAYER ON TOP OF IT AND IT SEEMS
THAT EVEN THESE FIBROBLASTS THEY
HAVE A DIFFERENT 8 HOURS NATURE
THAN THE 1S IN THE ALVEOLAR
SPACE.
>> YEAH, IMRAIT.
THERE IS A QUESTION NEERAJ, GO
AHEAD.
>> HERBERT EXCELLENT TALK, I
HAVE A QUESTION REGARDING WHY,
MAYBE I MISSED IT, YOU DIDN'T
SEE THE DOWN REGULATION IN COPD,
AND CRTAC IS A KNOWN PROGNOSTIC
MARKER FOR LUNG CARCINOMA MARKER
SO HOW DO WE DIFFERENTIATE
BETWEEN THE 2 PHENOTYPES.
>> YEAH, THANKS.
I GET MUCH MORE INTERESTED AGAIN
IN THIS PROTEIN NOW, WE SHOULD
HAVE CONTINUED TO STUDY IT, BACK
THEN BECAUSE IT'S SUPER
INTERESTING, BUT WE DON'T KNOW
MUCH ABOUT IT.
FORTUNATELY THERE WERE THEN,
SOME STUDIES ALSO IN THE COVID
CONTEXT AND SEVERAL APPLICATIONS
SHOWED IT'S THE MOST BIOMARKER
DOWN REGULATED OF THE 2% OF THE
NORMAL BASE LINE REMAINING
EXPRESSION IN SEVERE COVID.
AND WE CHECKED MANY DATA SETS,
SO IN COPD IT'S NOT REGULATED
AND YOU COULD THINK THAT BECAUSE
THE GRAPH I SHOWED YOU IS
TRANSCRIPTOMICS DATA FROM THE
GROUP AND QUITE HIGH NUMBER OF
PATIENTS, SO IT COULD BE SORT OF
A ARTIFACT THAT INDEED, IT'S
MAYBE DOWN REGULATED IN 82 BUT
THEN IT'S COMPENSATED SOMEHOW.
BUT WHEN YOU LOOK INTO SINGLE
CELL DATA, IT'S ALSO THERE, IT'S
NOT REALLY REGULATED.
SO IT MAKE ITS MUCH MORE
INTERESTING TO ME BECAUSE IT'S
SIGNIFICANT IN THIS CONTEXT.
>> HAVE YOU LOOKEDDA THE OTHER
LUNG DEC SUCH AS
[INDISCERNIBLE], THAT WOULD BE
INTERESTING TO SEE.
>> NO.
>> SO YEAH, IT SEEMS THERE'S
SOME RAP WITH COMPLEMENT
ACTUALLY SO IN ANOTHER PAPER WE
DISCUSS A DIFFERENT BIOMARKER,
CFHA 1 AND IT'S KNOWN CO
COMPLEMENT VECTOR H AND IT WAS
EXPRESSED BY [INDISCERNIBLE] AND
IT WAS NEGATIVELY CORRELATE WIDE
LUNG FUNCTION AND IT'S SORT OF
REGULATING THE COMPLEMENT
PATHWAY AND IT TURNS OUT WITH
THE LARGE SCALE STUDIES THAT IT
ISHT ACTING WITH ANOTHER PARTNER
IN THE COMPANY, AND REGULATORY
PATHWAYS.
SO IT SEEMS TO BE CONVERGING NOW
THAT BOTH BIOMARKERS WE FOUND
COULD ACT ON COMPLEMENT WHICH IS
KNOWN SINCE PROBABLY 30 YEARS,
THAT IT'S UPREGULATED AND
INCREASED IPS, TO COMPLEMENT
THIS CASE.
>> THANK YOU.
MSO ANOTHER QUESTION THAT'S KIND
OF --
>> SO ANOTHER QUESTION THAT'S
KIND OF IMPRESSED ME REALLY WAS
PEOPLE TALKING ABOUT PROTEOMICS
AND THEN THEY USE SINGLE CELL
RNA TRANSCRIPTOMICS AND YOU LOOK
AT THE SAME SAMPLES AND THE
CORRELATION IS REALLY LONG, A
LOT OF PEOPLE DISCUSSED THIS BUT
IN YOUR DATA CHA IS IMPRESSIVE
IS THAT YOU'RE TAKING LAVAGE
PROTEOMIC ANALYSIS AND THEN
LOOKING INTO SINGLE CELL RNA SEQ
FROM DISEASES INDIVIDUALS AND
ACTUALLY SEEING, YOU KNOW
CORRELATION SPECIFICALLY AS IT
RELATES TO DEC AND SO ON, AND --
DEC AND SO ON AND WHAT I
WONDERED WAS, AND YOU DID SOME
KIND OF AN AI ANALYSIS, I GUESS,
RANDOM FOR US OR SOMETHING, IS
THAT RIGHT IN SO IS THERE
SOMETHING THAT PEOPLE ARE MISS
NOTHING TERMS OF WHY PROTEOMIC
AND TRANSCREPT ORDER OF
MICRONSIC ARE NOT NECESSARILY
JIVING PROPERLY, I WONDER IF YOU
COULD COMMENT ON THAT.
YEAH, THANKS.
SO OBVIOUSLY NOT EVERYTHING
CORRELATES SO WE DID OFTEN IN
OTHER STUDIES AS WELL, WHEN YOU
DO TISSUE PROTEOMICS AND
TRANSKRPT ORDER OF MICRONSICS
AND CORRELATIONS, SO THERE'S
DIFFERENT WAYS HOW YOU CAN
CORRELATE THE DYNAMIC RANGE OF
THE PROAM I DON'T MEAN AND I
DYNAMIC RANGE OF THE TRANSCRIPT
THEMSELVES AND ANY ANYWAY THEY
DON'T CORRELATE WELL BUT IN THE
DIFFERENT CONDITIONS, COULD
ALREADY CORRELATE A BIT BETTER.
BUT THEN THERE ARE MANY TEAMS
THAT FORCE NICE REFLECTIONS LIKE
FOR INSTANCE ECM PROTEINS WILL
BE FOUND FOR COLLAGEN 4 IN THE
AGING CONTEXT, THE PROTEIN COULD
GET DOWN REGULATED WHILE THE
TRANSCREPT GETS UPREGULATED AND
VICE VERSA, BECAUSE OF SOME
FEEDBACK CONNECTIONS.
BUT HERE -- SO IN THIS
PARTICULAR CASE WE JUST TRY OUT
SORT OF, WE LEARN THAT SIGNATURE
IN THE PROTEOME, AND THEN WE
JUST ASK CAN IT ALSO PREDICT THE
DIRECT IN THE TRANSCREPT ORDER
OF MICRONSIC DATAY SOPHISTICATED
JUST BECAUSE IT HAS MANY MARKERS
IT COULD STILL WORK, NOT
DEPENDENT ON 1 PROTEIN AND IT'S
THE PLOT I SHOWED IN THE END FOR
THE PLASMA WHERE WE HAD ACTUALLY
MUCH LESS, AND LAVAGE WHICH IN A
WAY MAKES SENSE BECAUSE IT GETS
WAY MORE COMPLICATED YOU KNOW IF
THAT'S A PROBLEM OF HOW IN WHICH
WAYS IS SORT OF THE EXCHANGE
RATE BETWEEN TISSUE AND BLOOD
REGULATED WHICH IS DIFFERENT FOR
EACH AND EVERY PROTEIN.
SO THERE WAS A LOWER CORRELATION
AND THEN I'M ACTUALLY SHOWING
THE 1S THAT CORRELATE WELL, BUT
SOME DON'T CORRELATE WELL, SO
I'M SHOWING THE 1 DIRECTION HERE
WHICH IS GOOD.
>> GOT IT.
MAKES SENSE CARLA YOU GO AHEAD.
>> HI, THERE, CAN YOU HEAR ME
OKAY IN.
>> YES.
>> HI, THANK YOU ALL FOR THE
GREAT TALK IT'S REALLY NICE TO
BE PART OF THIS.
I JUST WANTED TO OFFER AND/OR
ASK ESPECIALLY SINCE YOU'RE
COLLECTING LAVAGE FLUID, WE HAVE
RECENTLY DEVELOPED A PROTOCOL TO
GROW EPITHELIAL ORGANOIDS FROM
LAVAGE FLUID AND SO, WE WOULD
LOVE TO COLLABORATE OR PURSUE IT
WITH YOU, WITH ANY OF YOU THAT
ARE COLLECTING LAVAGE FROM YOUR
PATIENTS SO THAT WE CAN SEE HOW
THE AIR WAY OR ALVERMEN INFECTED
O LIE EPITHELIAL CELL
COMOPPOSITE BEHAVIORIAL
PHENOTYPENTS ARE CORRELATING
WITH THE DISEASES THAT YOU'RE
STUDYING AND SOME OF THEIR
BIOMARKERS.
NYEAH, THAT SOUNDS EXCITING.
>> IS THAT WORKING WELL?
>> YES.
YEAH, IT ACTUALLY, WE PUSHLISHED
IT THIS SPRING IN THE
[INDISCERNIBLE] JOURNAL AND WE
CAN ESTABLISH AIRWAY ORGANOIDS
AND THEY CAN BE PASSAGED AND THE
ALVEOLAR ORGANIZED CELLS ALSO
CAN BE DERIVED AND THEY -- THEY
DON'T GO AS MANY PASSAGES,
THEY'RE A LITTLE MORE FINICKY
HOWEVER YOU DO GET PLENTY OF
GOOD ALVEOLAR CELLS TO WORK
WITH.
YEAH, HAPPY TO TALK MORE WITH
ANY OF YOU.
>> GREAT.
PLEASE GO AHEAD.
>> EXCELLENT TALK BY BOTH
SPEAKERS, I HAVE A QUESTION FOR
DR. GHOSH ABOUT THE MITOCHONDRIA
FUNCTION.
WHEN YOU LOOK AT THE 2 GROUPS
LOW AND HIGH, YOU SEE BASICALLY
BOTH MITOCHONDRIAOXIDATIVE
PHOSPHORYLATION AND DPLI COLSIS,
INCREASE IN THIS LOW, AND -- TD
CORRELATION OF THIS IN.
>> NO WE DID NOT LOOK AT
ABSOLUTE LEVEL AND BASICALLY
THAT'S WHAT WE WERE EXPECTING
WHEN WE SAW THE MORE OX PH OS,
SO WHEN THEY'RE NOT DOING
ANYTHING, THEY'RE TBLI COLSIS
DEPENDENT AND WHEN THEY START
SENSING INJURY THEY WOULD GO TO
OX FIRST.
IN OUR CASE WE WERE SEEING BOTH
HIGH OX AND FLOX WE DIDN'T GO
AND ACTUALLY, OUR PLAN IS TO USE
LIKE STABILIZED ISOTOPE AND GET
AND TAKE OUT THE DEEPER DIVE
INTO IT BUT AT THIS POINT ALL WE
KNOW IS THAT THE HIGH ATP IS
COMING FROM BOTH WAY UP MAKING
AND [INDISCERNIBLE].
SO YOU'RE SEEING LIKE THIS --
OKAY, THANK YOU.
THANKS.
>> MOUMITA, I HAVE A QUESTION
FOR YOU, IT'S FASCINATING HAVE
YOU THIS SUSCEPTIBILITY OF THE
LARGE AIRWAY AND SHOWING THE
LUNG FUNCTION CORRELATION.
AND I WONDERED, 2 THINGS, I MEAN
IS IT POSSIBLE THAT ESSENTIALLY
THESE SUSCEPTIBLE INDIVIDUALS OR
WHAT YOU'RE THINKING RELATED TO
THAT, I WOULD SAY, YOU KNOW THAT
ESSENTIALLY THE ENTIRE LUNG IF
YOU WILL BECAUSE IT'S EXPOSED TO
CIGARETTE SMOKE IS SUSCEPTIBLE.
I MEAN WE'RE -- YOU KNOW WE
THINK ABOUT SMALL AIRWAY
EPITHELIAL CELLS, YOU KNOW CLUB
CELLS FROM THE SMALL AIRWAY,
BECOMING YOU KNOW TYPE 2 CELLS
IF THEY'RE NOT FUNCTIONING,
SOMETHING HAPPENS TO THE PATHWAY
GIVES RENK MA, BUT IN YOUR CASE
EVERYTHING'S NOT WORKING
ESSENTIALLY SO WHAT YOU DO YOU
THINK ABOUT THAT?
>> THAT'S EXACTLY WHATY WOO ARE
THINKING AND IN A COMPLETELY
DIFFERENT COHORT WHERE WE'RE
LOOKING AT SECOND HAND SMOKE
EXPOSE AND YOU ARE LOOKING AT
SIGNAL NAISAL BRUSHINGS AND
GETTING BASAL PROGENITOR FROM
NAISAL BRUSHING WE SEE SIMILAR
THING.
WE SEE LOSS OF PROGENITOR
FUNCTION AND CORRELATION, WITH
TOTAL LUNG CAPACITY, SO THATIA
HAPPENING IN THE NOSE, TO LUNG
FUNCTION.
SO, IT IS REALLY IT SEEMS THAT
AS THE WHOLE LUNG SMOKE FIRST OR
SECOND HAND THAT EVERYTHING,
LIKE YOU KNOW THIS PROGENITOR
DYSFUNCTION IS HAPPENING ALL THE
WAY, STARTING FROM NAISAL
BRUSHING TO, THAT'S WHY IN THE
CURRENT STUDY WE ADDED NAISAL
BRUSHING BECAUSE IT WOULD BE
EASIER TO COLLECT THAN DOING
BRONCHIO SCOPEY BUT WE WANT TO
DO BOTH NONAPOPTOTIC MAKE SURE
THAT YOU KNOW NORMAL CASE FOR
NAISAL REPRESENT BRONCHIAL BUT
IN TEASE WE DO NOT KNOW WHETHER
THAT CHANGES, SO THAT'S REALLY
THE PLAN, IF THEY HAVE BEHAVE
SIMILARLY, THEN IT WOULD BE
EASIER, THESE PEOPLE DO NOT HAVE
A DISEASE.
IF WE WANT TO COLLECT CELLS FROM
THEM, DOING NAISAL BRUSHING WILL
BE MUCH MORE PEOPLE WOULD BE
MUCH MORE WILLING TO COME AND DO
THAT RATHER THAN DOING A BRONCH
COSCOPY.
NRIGHT, IN EMERGING ITS OF WHAT
WE TALKED ABOUT SUSCEPTIBILITY,
RESILIENCE AND SO ON, I THINK
OBVIOUSLY THESE PEOPLE ARE
SUSCEPTIBLE, THERE MUST BE
SUSCEPTIBILITY GENES AND WE DO
SEE THAT AS WELL, I MEAN MUCUS
IN THE AIRWAY, TRANSLATES IN THE
NOSE AS WELL FOR EXAMPLE WHEN
THAT HAPPENS, JOSE, YOU HAD A
QUESTION?
>> YEAH, IT WAS A COMMENT ON
WHAT MOUMITA WAS SAYING, I THINK
IT'S INTERESTING YOU RAISED IN
IDEA OF USING NAISAL PHARYNGIAL
SWAPS AND THAT YOU'RE SEEING
THIS NICE CORRELATION BETWEEN
WHAT'S HAPPENING IN THE VERY
UPPER AIRWAY AND WITH THE LOWER
LUNG FUNCTION.
WE'VE SEEN IN SOME OF OUR
STUDIES PARTICULARLY WITHIN THE
REALM OF COVID-19 THAT YOU CAN
ACTUALLY HAVE VERY INVERSE
EFFECTS IN TERMS OF HOW DRIVER
THAT YOU PICK UP IN THE NAISAL
MUCOSA, OF SEVERE DISEASE, MAY
ACTUALLY BE THEN DETRIMENT OF
MAY BE BENEFICIAL IN LOWER
AIRWAY AND VICE VERSA, RIGHT IN
SO I THINK THAT EACH DEC IS
LIKELY TO HAVE IT'S OWN
RELATIONSHIP, AND SO, WE SHOULD
MAYBE START TO THINK ABOUT
CATALOGING THESE BECAUSE OFTEN
TIMES THERE'S THE -- YOU KNOW
HOW HIGH CAN I GOT TO UNDERSTAND
WHAT'S HAPPENING IN THE LOWER
COMPARTMENT, AND CLEARLY BASED
-- YOU'VE THOUGHT ABOUT THIS
ISSUE AS WELL, BUT I THINK COULD
BE AN INTERESTING OUTPUT OF WHAT
WE'RE DISCUSSING HERE.
>> I COULD NOT AGREE MORE
BECAUSE ALSO THERE IS, LIKE WITH
[INDISCERNIBLE] DIFFERENT THINGS
WE ARE DOING, I DO BELIEVE THERE
ARE DIFFERENCES IN BRUSHINGS,
THE BASAL CELLS, COLLECTED BY
BRUSHINGS AND BASAL CELLS
COLLECTED BY BIOPSIES BECAUSE
THOSE CELLS ARE REALLY TOUCHING
BASAL MEMBRANE.
WHEN YOU ARE TALKING ABOUT STEM
CELLS, PROGENITOR CELLS THERE IS
A SIGNIFICANT DISRCHES BETWEEN
THEM AND SO, AND PROBABLY THAT
BECOMES MORE ABERRANT WHEN
LOOKING AT DISEASE VERSUS
HEALTHY SOPHISTICATEDY THERE ARE
A LOT MORE NUANCED THING GOING
ON HERE THAN JUST LIKE
COLLECTING FROM ANY COMPARTMENT.
>> SO WE HAVE 1 LAST QUESTION
BEFORE THE TOP OF THE HOUR,
MICHA, GO AHEAD.
>> SO, YEAH THIS, IS FOR
DR. SCHILLER, AND I LOOK HOW YOU
OPENED YOUR TALK ABOUT AS
TALKING ABOUT TISSUE STATE AS A
CONCEPT AND I GUESS MY QUESTION
IS SORT OF HOW DO WE GO ABOUT
FROM STUDYING IT TO -- LIKE WHAT
DOES A THERAPEUTIC TARGET LOOK
LIKE AT THAT SYSTEM SCALE, DO WE
HAVE TO HIT MULTIPLE CELL TYPES
IN MULTIPLE WAYS, ARE YOU GOING
TO BE ABLE TO FIND SMSK CONTROL
POINTS THAT WILL PUSH THIS
AROUND BECAUSE WE'RE LOOKING AT
MULTIPLE LINEAGES CHANGE NOTHING
MULTIPLE WAYS AND COORDINATIONS
AND TALKING TO EACH OTHER IN
DIFFERENT WAYS SO IF THAT'S A
THERAPEUTIC TARGET, HOW DO WE
THINK ABOUT THAT CLINICALLY OR
FROMAGE ENGINEERING STANDPOINT.
WHAT DO YOU DO WITH THAT?
>> YEAH, THAT'S A GREAT
QUESTION.
THANKS.
INDEED MY INTUITION WOULD BE WE
NEED THESE COMBINATION
THERAPIES, SO THAT'S WHY SORT OF
INDICATE HOPEFULLY WE CAN HAVE A
WAY OF NOT DOING EACH AND EVERY
EXPERIMENT, RIGHT?
LIKE EACH AND EVERY COMBINATION,
THAT'S WHAT SEEMINGLY SOME OF
THESE, SOME OF US WOULD BE GOD
IN PREDICTING COMBINATIONS AND
WE CAN GO BACK TO THE LAB AND DO
THE ACTUAL EXPERIMENT.
YEAH, AND THEN, YOU KNOW WHETHER
THIS -- HOW EASY IT IS TO GO TO
CLINICAL STUDIES SETTING WITH
COMBINATION THERAPIES, NOT SURE
ABOUT I THINK THAT'S PROBABLY
WHERE MOST SUCCESS COULD COME
FROM BECAUSE IT WAS A QUESTION
TODAY ABOUT ALL THESE DIFFERENT
MARKERS OF THIS TRANSITION
STATES FOR EXAMPLE, YEAH, SO YOU
HAVE SO MANY TACKERETS RIGHT?
AND THAT'S ONLY 1 LINEAGE.
BUT I GUESS THERE ARE THESE
CONTROL POINTS, LIKE A CERTAIN,
LIKE, THE [INDISCERNIBLE]
FACTORS IS 4 CHAINS AND IT'S
DOING A LOT, RIGHT?
SO ...
>> GREAT.
THANKS.
>> THANK YOU FOR THE
DISCUSSIONS, I THINK DR. LU, YOU
WANT TO CLOSE, I WILL HAND IT TO
YOU.
NYEAH, SURE, THANK YOU SO MUCH,
GREAT SECTION, I THINK WE ALL
CAN AGREE, WE HAD A GREAT,
GREAT, MEETING TODAY.
SIGNIFY CERTAINLY LEARNED A LOT
AND HUGE THANKS TO ALL OUR
CO-CHAIRINGS AND CO MODERATORS
AND ALSO SPEAKERS.
LOOKING FORWARD TO HAVING
ANOTHER EXCITING DAY TOMORROW.
SO TOMORROW WE ARE STARTING AT
10:30.
LIKE TODAY, SO SPEAKERS AND
MODERATORS MAY JOIN AROUND
10:00 O'CLOCK SO WE CAN TEST
YOUR AUDIO AND VIDEO.
AND I'M GOING TO CLOSE THE
MEETING TODAY.
HAVE A GREAT EVENING.
THANK YOU.
>> THANK YOU.
>> THANKS.