>> WELCOME EVERYONE TO THE
WEDNESDAY AFTERNOON LECTURE
SERIES, I'M DR. KELLY, I AM A
CCR DEPUTY DIRECTOR AND I AM
VERY EXCITED TO INTRODUCE
TODAY'S SPEAKER DR. HANS CLEVERS
WHO HAS PIONEERED THE FIELD OF
ORGANOIDS
OUR SPEAKER TODAY TRULY IS
WELL KNOWN TO MOST OF YOU, HANS
CLEVERS HAS LONG HELD THE
POSITION OF PROFESSOR OF
DEVELOPMENTAL BIOLOGY, IN
MARCH 2022 HE ACCEPTED THE
POSITION OF HEAD OF ROCHE PHARMA
RESEARCH AND EARLY DEVELOPMENT
INDEED, HE'S GIVING HIS LECTURE
TODAY FROM BOSUL WHERE IT IS NOW
9 P.M. SO DR. CLEVERS OBTAINED
HIS MD DEGREE IN 1984 AND
Ph.D. IN 1985 FROM HUBRECHT,
UNIVERSITY, AND HIS POST
DOCTORAL WORK IN 86-89 WAS DONE
AT THE DANA FARBER CANCER
INSTITUTE AND HARVARD,
UNIVERSITY.
THROUGHOUT HIS CAREER, HE HAS
WORKED ON THE ROLE OF SIGNALING
AND STEM CELLS IN CANCER AND HIS
DISCOVERIES INCLUDE TCF AS THE
NUCLEAR WINTER WEATHER EFFECTOR,
THE LOCAL OF WINTER WEATHER IN
STEM CELL BIOLOGY AND OF WITN
COLAN CANCER AND LGR IS A MARKER
OF NOVEL TYPES AND THE AMPLIFIED
ARE OUR RESPONDENTS AND NEW
METHODS TO GROW EVERYEXPANDING
ORGANOIDS DERIVE FRIDAY A RANGE
OF HEALTHY AND DISEASED HUMAN IS
SHE.
THIS WORK LED TO 750
PUBLICATIONS AT LEAST AND 90,000
CITATIONS.
DR. CLEVERS IS A MEMBER OF THE
ROYAL ACADEMY OF ARTS AND
SCIENCES, AMERICAN ACADEMY OF
THE ARTS AND SCIENCES U.S.
AMERICAN ACADEMY OF ARTS AND
SCIENCES AND FRENCH ACADEMY OF
TOMORROWS AND HE IS A RECIPIENT
OF AND ORGANOIDS TO MODEL HUMAN
DISEASE, HAN SWELCOME TO THE NIH
AND THANK YOU SO MUCH FOR BEING
HERE AND WE LOOK FORWARD TO YOUR
TALK.
>> THANK YOU VERY MUCH VERY MUCH
KELLY, FOR THE INTRODUCTION AND
SPEAK
WHAT I WILL TRY TO DO IS GIVE
A BIT OF BACKGROUND AS TO HOW WE
MORE OR LESS BY CHANCE
DISCOVERED THIS TECHNOLOGY TO
GROW HUMAN ORGANOIDS FOR HUMAN
AND DISEASE AND TAKE IT BACK 25
YEARS OR SO AGO, WHEN MY LAB
JUST DISCOVERED THAT THE TCF
TRANSCRIPTION FACTORS ARE THE
EFFECTORS OF THE PATHWAY AND AT
THE SAME TIME WE NOTE THAD WHEN
WE KNOCK OUT 1 OF THOSE TCF 4,
THAT THE PROCESS YOU'RE LOOKING
AT COMES TO AN ABRUPT HALT AND
THIS IS TO SEE THE STEM CELLS IN
ACTION AND WE ALSO THEN SHOWED
THAT IN COLOON CANCER IN
PARTICULAR, ACTIVATING THE
MUTATIONS ACTIVATE THE STEM
CELLS, LEADING TO ADENOMA
PROLIFERATION.
HERE YOU SEE VILLUS IN THE HUMAN
BEING AND THESE ARE ABOUT 6-10
AND THIS IS WHERE IT WAS WELL
KNOWN THAT STEM CELLS AND THE
PROCESS SO CELLS ARE BORNE NEAR
THE BASE OF THE CRYPT FROM THESE
ELUSIVE STEM CELLS, THE DAUGHTER
CELLS ONLY TAKE 2 DAYS WHILE
THEY PROLIFERATE TO XIPT THE
CRYPTS, THEN THEY TAKE IT
RAPIDLY 1 OF 10 DIFFERENT CELL
TYPES AND CELL FETES, CELLS ARE
LIKELY TO SETTLE AT THE BASE,
YOU SEE THESE LARGE CELLS HERE,
ALL OF THE CELL TYPES KEEP ON
MOVING UP AND WHEN THEY'RE ABOUT
5-6 DAYS OLD IN THE MOUSE, THEY
REACH THE TIPS OF THE VILLI AND
THE APOPTOSIS AND A MOUSE IN THE
TISSUE CELLS EVERY WEEK OR SO IN
HUMANS IT MIGHT BE CLOSER TO 2
WEEKS, PROBABLY THE FASTEST AS I
SAID, CELL RENEWING TISSUE.
SO WE HAVE FOUND THAT IN CANCER
AND IN PHYSIOLOGY OF CRYPTS,
THIS IS THE DRIVING FORCE, IT
TOOK US A WHILE TO FIND OUT THE
TARGETS PROGRAM, WE HAD TO WAIT
UNTIL MICROARRAY WAS INVENTED IN
THE EARLY 2000S.
NOW YOU WOULD PROBABLY DO THAT
WITH RNA SEEK, THIS ALLOWED US
IN COLON CANCER CELL LINES,
ABOUT 200,000 GENES THAT WERE
UNDER THE CONTROL OF THE THIS
WEN PATHWAY AND WE FOUND THAT
THIS IS ACTIVE IN CRYPTS AND
THEN WE THOUGHT POSSIBLY AS NICK
BARKER A BRITISH POST DOC FOUND
IT HIDDEN IN 200 TARGET GENES
AND MOST OF THEM ACTUALLY
EXPRESSED IN THE ENTIRE CRYPTS
AS THEY WERE EXPRESSED IN THE
ENTIRE ADENOMA CARCINOMAS OF THE
GUT AND THAT HIDDEN IN THERE,
THAT MAYBE 1 OR MORE THAT WOULD
MARK THE STEM CELLS AT THE BASE,
AND YOU SEE A GENE THAT PICKED
AFTER SEVERAL EXERCISES EARLIER
ON, AND THIS IS A KNOCK OUT AND
GFP BUT ALSO A RECOMBINATION EFR
AND I WILL SHOW THIS IN THE NEXT
SLIDE.
THIS LIGHTS UP TINY CELLS, THIS
IS 1 CELL ON THE LIGHT, THE
LARGER CELLS ARE THE BANNER
CELLS, POST MITOTIC CELLS, FULLY
DIFFERENTIATED, ACTIVE IN
KEEPING IN OUT OF THE CRYPTS AND
THESE CELLS WE THEN THOUGHT THEY
MIGHT BE GOOD CANDIDATES TO BE
STEM CELLS.
THEY'RE AT THE BASE OF THE
CRYPT, THEY'RE PROLIFERATIVE AND
INDEED WHEN WE NOW PERFORM A SO
CALLED LINEAGE TRACING
EXPERIMENTS, HERE WE CROSS THIS
MOUTH WITH A GOOD REPORTER
AND--I WON'T GIVE YOU TOO MUCH
DETAIL HERE BUT WE CAN NOW TERM
1 OF THESE CANDIDATE STEM CELLS
PERMANENTLY PLURIBU AND WHEN THE
CELL DIVIDES THE DAUGHTER CELLS
WILL BE BLUE AND YOU SEE HERE
SUMMARIZING THE EXPERIMENTS OF
MYC, THE FIRST FEW DAYS YOU SAW
BLUE DAUGHTER SALES ARISING FROM
THIS MARKED STEM CELL IN ADULT
MOUSE, AFTER 2 OR 3 DAYS THEY
XIPT THE CRYPTS, THEY
DIFFERENTIATE OUT INTO 1 OR MORE
OF THESE CELL TYPES, GOBLETS,
ENTHUSIASMER O CITES AND DAY 3
OR 4 OR 5 OF THE EXPERIMENT AS
HOPED FOR, WE SAW THIS BLUE
RIBBON SLOWLY CREEPS UP THE
FRANKS AND THEN BY DAY 5 OR 6 OF
THE EXPERIMENT, THE FIRST BLUE
CELLS ARRIVE AT THE TIPS OF THE
VILLI AND THEY UNDERGO
APOPTOSIS.
NOW IF YOU DON'T TERMINATE
EXPERIMENT HERE BUT WE LEAVE THE
MICE FOR ANOTHER 2 YEARS THAT
BLUE RIBBON IS STILL THERE, SO
THE CELL WE MARKED AT THE START
OF EXPERIMENT IS LONG LIVED
EVERY DAY IT KEEPS ON CRANKING
OUT THESE BLUE DAUGHTER CELLS,
2IERS LONG, 3 YEARS LONG, AND
WHEN WE DO THE EXPERIMENT WELL
AND WE ONLY MARK HERE AND THERE
A SINGLE CELL AND PRODUCE THESE
RIBBONS WE CAN HAVE READ OUT IN
THE RIBBONS WHAT KIND OF CELL
TYPES CAN BE PRODUCED BY THIS
CELL AND WHAT WE FOUND, WHAT
NICK FOUND WAS THAT ALL THE
OTHER CELL TYPES, THAT YOU SEE
IN THE EPITHELIUM, ARE PRESENT
IN 1 OF THESE COLONIAL RIBBONS
AND IMPLAYING FITTER THAT THE
CELL MARKER IS LONG LIVED UNLIKE
A LOT OF CELL TYPES OR THAT
SECOND THAT ACTUALLY IS
MULTIPOTENT STATANT BECAUSE IT
MAKES ALL CELLS ON THIS TISSUE.
NOW WE TRY TO PUBLISH THAT THESE
CELLS ARE THE CELLS IN THE GUT
AND IT'S A LOT OF RESISTANCE
BECAUSE IT'S GENERALLY ASSUME
THAD THESE CELLS DIVIDE VERY
RARELY AND CHARACTERISTIC OR
DEFINING CHARACTERISTIC OF AN
ADULT STEM CELL WOULD BE THE
FACT THAT IF YOU LABELED ITS DNA
THAT CELL WOULD HAVE RETAINED
THE DNA LABEL FOR LONG PERIODS
OF TIME BECAUSE IT WOULD NOT
DIVIDE FOR A LONG PERIOD, THAT
WAS QUITE A REASONABLE
HYPOTHESIS, BECAUSE, NO, WHEN
YOU DIVIDE, YOU HAVE TO COPY DNA
AND MAKE MISTAKES SO STEM CELLS
MIGHT NOT WANT TO DO THAT,
HOWEVER, THE GUT STEM CELL, YOU
KNOW NOW KNOW THE STOMACH STEM
CELL, GO THROUGH MANY, MANY CELL
CYCLES IN A MOUSE WE THINK THESE
CELLS GO ABOUT THROUGH ABOUT A
THOUSAND CELL CYCLES IN A
LIFETIME OF MOUSE, CONSECUTIVE
MICE WITH 20,000, BUT MUST BE
ENORMOUS BUT SOMEHOW THESE CELLS
MANAGE.
NOW BASED ON THIS, ANOTHER POST
DOC IN THE LAB, YOU SEE HIM
HERE, THEY SAT DOWN AND WE ASKED
CAN WE POSSIBLY CREATE AN
ENVIRONMENT IN A PLAOF THE IRK
DISH, IN A PET RI DISH THAT
MIMICS THE ENVIRONMENT AND THE
INTENTION WAS NOT WHAT YOU SEE
HERE, THE INTENTION WAS TO SAKE
A SINGLE STEM CELL, TOP LEFT AND
THE GROWTH FACTORS WE KNOW THAT
WIND WAS KEY, WE KNEW ALSO THAT
OUR RESPONDENT IS AN AMPLIFIER
THE SIGNALS WE DIDN'T KNOW IT
WAS THE LIGAND OF OUR LGFR
RECEPTOR BUT ACTUALLY THAT WE
FOWBD OUT MUCH LATER AND
EXPLAINS WHAT WE WERE SEEING, SO
WE HAD OUR [INDISCERNIBLE] IN
OUR COCKTAIL AND ACTIVATOR OF
THE KINASE RECEPTOR AND THEN
FROM AN OLD TRANSGENIC MOUSE
EXPERIMENT, WE KNEW WE HAD TO
BLOCK BMP SIGNALS, IF YOU
EXPRESSION NOGGIN IN THE GUT OF
A MOUSE, YOU WILL SEE MANY, MANY
MORE CRYPTS SO IT WILL BE
INHIBITION SOMEHOW CREATES A
CRYPT FROM THIS ENVIRONMENT.
NOW THE COMBINATION OF THESE
GROWTH FACTORS, NO SERUM, BUT WE
DO THIS IN MATE RIGEL AS
PIONEERED BY [INDISCERNIBLE]
OVER THE PAST 30 OR 40 YEARS,
WAS A MIX OF LLAMA NIN,
MATRIGEL, AND IT WAS TO CREATE
MANY STEM CELLS FROM A SINGLE
STEM CELL LIKE PEOPLE ARE
GROWING ES AND IPS CELLSA THE
TIME BUT WE GOT SOMETHING VERY
DIFFERENT.
WE GOT EPITHELIAL STRUCTURES AND
KD--SALLY TOSHIRO, STARTED
LOOKING QUICKLY AND THERE WOULD
BE MULTIPLE STEM CELLS AT THE
BASE, AND OTHER CELLS THAT ARE
THERE, THE OTHER CELLS IN THIS
WILL BE RAPIDLY DEFINING, AND A
CENTRAL LUMEN WAS LINED BY ALL
THE OTHER 9 OR 10 CELL TYPES
THAT WE REMOVE FROM THE PRIMARY
EPITHELIUM AND A NUMBER OF YEARS
LATER BY SEQUENCING WE
DISCOVERED 2 NOVEL CELL TYPES
THAT WE HAVE NOT SEEN EARLIER IN
ORGANOIDS, WE THEN WENT BACK TO
A REAL GUT AND WE FOUND THAT
THEY REALLY EXISTED.
SO IT'S REALLY REMARKABLE THAT A
SINGLE STEM CELL, 3 GROWTH
FACTORS, MATE RIGEL AND OTHERS
APPARENTLY REMEMBERS HOW TO
BUILD A COMPLETE VERSION OF THE
TISSUE THAT EMERGES FROM.
AND TOSHIRO CALLED THESE MANY
STRUCTURES AND TODAY THEY WOULD
BE CALLED DECIMAL ORGANOIDS.
SO THEY GROW FOR VERY LONG
PERIODS OF TIME AND 10 YEARS AND
MOUSE LIVES ONLY 3 YEARS AS YOU
PROBABLY KNOW, TELOMERES STAY
LONG, WE'VE NEVER SEEN
SENESCENSEL BY SEQUENCING, WE'VE
NEVER SEEN ONCOGENIC MUTATIONS
AND PHENOTYPICALLY THEY STAY
STABLE OVER TIME AND THE
ULTIMATE EXPERIENCE TO SEE IF
THEY REALLY STAYED NORMAL WOULD
BE A TRANSPLANTATION, AND THIS
WAS DONE BY TOSHI IN MY LAB AND
[INDISCERNIBLE] AND THE PEOPLE
TOKYO, SO TOSHI, TOOK A SINGLE
STEM CELL FROM THE MICE WHERE IT
WAS GREEN BUT THE ENTIRE MOUSE
WAS RFP POSITIVE SO IT WAS RAT
FLUORESCENCE, AND IT ALLOWED US
TO KEEP TRACK OF THE TISSUE
WE'RE GROWING SO FROM A DUTCH
MOUSE ISOLATED TO SINGLE CELL,
UP TO ABOUT A HUNDRED MILLION
CELLS IN THE FORM OF THIS MANY
AND THERE AND THE PEOPLE HAD TO
TREATED SEVERAL DOZEN MICE WITH
DSS, WHICH IS A CHEMICAL THAT
INDUCES INFLAMMATORY BOWEL LIKE
SYNDROME IN THESE MICE.
THEY INFUSE THESE RED
FLUORESCENT DUTCH ORGANOIDS
THROUGH THE ANUSES OF THESE
SEVERAL DOSEN MICE AND OVER THE
NEXT HOUR THIS MOVIE SUMMARIZES
THE FLOAT AROUND, ALWAYS THE
BASAL SIDE, SO THE INTERNATIONAL
CLASSIFICATION GRENS ARE ON THE
OUTSIDE AND THE MOMENT THEY SEE
THESE OPEN LESIONS, OF
COLLAGENS, FIBER NECT
EPIGENETICS THEY WILL ATTACH AND
OPEN UP, LIKE A LIVING BAND 8,
ORGANOIDS AND I EXPRESSED THEY
GROW FROM A SINGLE STEM CELL, A
FEW MONTHS EARLIER, THESE WILL
SEAL THESE LESIONS AND THIS IS
THE REAL EXPERIMENT, WITH THESE
MICE, WHAT IS BLACK IS THE
JAPANESE TISSUE, HERE YOU SEE
THE DUTCH TRANSPLANTED TISSUE, I
STRESS AGAIN GROWN FROM A SINGLE
TRAN PLANT CELL USED TO GROW
MAYBE 40 MICE AND YOU SEE THE
LARGE PATCHES OF RED FLUORESCENT
TISSUE SO THE OFFSPRING OF A
STEM CELL THAT WAS ISOLATED AND
GROWN, FULLY INTEGRATED IN THE
TISSUE AND THE ONLY WAY YOU CAN
FIND IT IS WITH A CONFOCAL
MICROSCOPE AND ANY MARKER
ANALYSIS WE DO IS NORMAL TISSUE.
NOW 1 PREDICTION WOULD BE THAT
THE STEM CELL SHOULD BE AN
EXHAUSTED BY NOW BECAUSE IT'S
EXPANDED SO DRAMATICALLY
OVERTIME IN THE LAB.
SO WE LOSE THESE PATCHES, THAT
IS NOT WHAT'S HAPPENING, OTHER
PREDICTION WOULD BE FROM PEOPLE
WHO BELIEVE THE STEM CELL SHOULD
REALLY BE QUIESCENT IS THAT
THESE CELLS PROBABLY NOT ARE
SOMEHOW TRANSFORMED SO IT MIGHT
SEE NEOPLASMS, ALSO WE NEVER SAW
THESE THINGS HAPPENING.
AND BASED ON THIS, IT HAS GONE
ON TO DO THIS WITH HUMAN
ORGANOIDS TO IMMUNO DEFICIENT
MICE, THEY ALSO INTEGRATE NICELY
AND CURRENTLY PERFORMING A FIRST
IN MAN TRIAL TREAT INFLAMMATORY
BOWEL DISEASE WITH PATES FOR
THERAPY RESISTANT FORM OF IBS
AND IT TREATS THEM WITH
ORGANOIDS GROWN FROM HEALTHY
PARTS OF THEIR OWN COLON SO THIS
WOULD BE AN AHATOLOGIOUS
TRANSPLANT AND OF COURSE, WE ARE
CURIOUS TO SEE IF THIS WILL WORK
OUT.
THIS IS PROBABLY ALL
EXPERIMENTAL IS ABOUT 10 YEARS
OLD, BUT WE FIRST THOUGHT THAT
WAS UNIQUE TO THE GUT STEM
CELLS, BECAUSE THEY'RE
INCREDIBLE LOAMACYY ACTIVE AND
THEN THIS SHOWED THIS COULD BE
DONE FOR PANCREAS AND IT'S NOT
ONLY PROLIFERATIVE TISSUE, UNDER
NORMAL CONDITIONS BUT YOU SEE
THERE NOW A LONG LIST OF
PROTOCOLS AVAILABLE IN HUMAN AND
MOUSE, WHERE YOU CAN START WITH
A TINY BIT OF TISSUE FROM A
BIOPSY FOR INSTANCE, CAN YOU
SEND THEM AROUND THE WORLD AS
LONG AS YOU KEEP IT ON ICE, ON
0-DEGREES IN A COMPLETE MEDIUM,
CAN YOU SEND THEM AROUND FOR 2
OR 3 DAYS BEFORE YOU PUT THEM IN
CULTURE.
THEY WILL NEED AN ACTIVATOR ON
THE WIN PATHWAY, OVER THE KINASE
RECEPTOR AND THEY TYPICALLY NEED
INHIBITORS OF TGF BETA AND
B-SIGNALING IN ADDITION TO THE
MEMORY LIKE ESTROUGH ATOM GENS,
PROSTATE, HERE IT IS LIKE
TESTOSTERONE, SO THERE ARE
ADDITIONAL COMPONENTS.
MANY OF THESE CULTURE CONDITIONS
HAVE BEEN PUBLISHED, AND THEN WE
CAN DO BASICALLY GET YOUR
TISSUE, GRIND IT UP IN SMALL
PIECES, STICK IT IN, PUT IN THIS
MATE RIJELL AND AND YOU SEE THAT
VERY RAPIDLY CLOSE ENOUGH AND
THE STRUCTURE AND THEN THEY GOT
IT GROWING IN HERE AS WELL AND
DEPENDING ON THE CONDITION, YOU
WILL SEE STEM CELLS THAT ALL THE
DIFFERENTIATION, AND ALL SORTS
OF BASIC SCIENCE QUESTIONS OR
USING KNOCK OUT MICE OR MAYBE AT
LEAST CELL LINES, AND ALSO CAN
BE GROWN DIRECTLY FROM IN HERE
AND CANCERS WE'VE DONE A LOT ON
CYSTIC FIBROSIS AS WELL, SO YOU
CAN REALLY MEASURE AS WELL, THE
DRUG RESPONSE, AND AND RESPONSE
TO THE DRUG, ALSO THE
INTERACTION OF THE DRUGS, ALSO,
GIVE SOME EXAMPLES FOR
INFECTIOUS DISEASES QUITE WELL,
THEY ARE THE HEPATOCYTES FROM
THE HUMAN LIVER AND AS YOU
PROBABLY REALIZE, THESE ARE VERY
IMPORTANT CELLS IN DRUG
DEVELOPMENT PROCESS, BUT ALSO IN
THE STANDS TO CONTROL METABOLISM
AND THE SIPGHT SIS OF MANY SERUM
PROTEINS, ET CETERA, ET CETERA,
THEY WILL DELIVER ESSENTIALLY
ONLY HAS 2 REAL LIVER CELL
SITES, THAT BUILD THE BILE DUCTS
AND ALL THE OTHER CELLS THAT SEE
THE COOPER CELLS,
[INDISCERNIBLE] CELLS ARE
MIGRATING INTO THE LIVE WHEN
IT'S FORMING BUT THE IT'S THE
[INDISCERNIBLE] THAT BUILD THE
LIVER, THE LIVER IS PROBABLY THE
MOST REGENRATIVE ORGAN OF THE
HUMAN BODY AND THERE'S 2 WAYS BY
WHICH IT CAN REPAIR THE CELLS
BUT ALL HEPATOCYTES ARE SICK FOR
AN INFECTION OR INTOXICATION,
THE BILE DUCT CELLS CAN BE
DIFFERENTIATED TO OVAL CELLS,
PROLIFERATE AND THEN RECREATE
BILED CELLS AS WELL AS
HEPATOCYTES AND THEN METASTASIS
FROM COLAN CANCER, IF YOU LIVE
THE LIVER IN PLACE ISSUES THE
LIVER WILL RAPIDLY GROW BACK AND
THEN THIS IS DONE BY HEPATOCYTES
BECAUSE REMAINING HEPATOCYTES
ARE HEALTHY, CAN YOU DIVIDE
THOSE SO YOU WOULD NEVER CALL
THE REAL STEM CELLS, YOU HAVE TO
CREATE A LOST TISSUE AND THEN IN
THE RESTORING LIVER, YOU WILL
SEE HEPATOCYTES, YOU WILL SEE
THE BILE DUCT FORMED AS WELL
FROM THIS, AND YOU WILL SEE THE
OUTER CELLS MIGRATING.
SO HERE YOU CAN SEE THAT THE
COLINGIOO CITES THEY'RE FULLY
DIFIENTIATED GLANDIO CITES AND
PUT IN THE RIGHT COCKTAIL GROWTH
FACTOR, MAYBE FROM MOUSE A BIT
EARLIER ABOUT YOU THIS IS HUMAN
HERE WHAT YOU SEE, ALMOST EVERY
COLANGIO CITE ORGANOID, THESE
WILL HAVE EARLY COLANGIOCITE
MARKERS IN VITROW, WE CAN
DIFFERENTIATE TOWARDS IT BUT IT
NEVER GET VERY ADULT LIKE, THEY
REALLY ARE IMMATURE, AND YOU
HAVE TO TRAN PLANT TO SEE THEY
CAN MAKE HEPATOCYTES AND HERE
ARE LARGE CELLS, BEAUTIFUL
CELLS, YOU CAN BUY THEM, THERE
ARE OFTEN USED IN THE DRUG
DEVELOPMENT INDUSTRY, BUT IT'S
BEEN ALMOST IMPOSSIBLE TO KEEP
THESE HEPATITIS EAT O CITES IN
THEIR HEALTHY STAGE FOR MORE
THAN A FEW DAYS OR A WEEK.
THERE'S REALLY A CELL TYPE TO
CANCER CELL LINES THAT MANY LABS
USE ARE CARCINOMAS, OF LIVER
CANCER, AND THEY--THEY EXPRESS
THE LIVER [INDISCERNIBLE] WILL
TYPICALLY BE A HUNDRED OR
THOUSAND FOLD LOWER THAN A REAL
HEPATITIS EAT O CITE, THE GROWTH
WOULD BE THE SAME.
SO WE WORKED VERY HARD FOR
MULTILE YEARS UP UNTIL THE
CHINESE POSTDOC OUR OWN LAB BACK
IN CHINA CAME UP WITH A
COMPLICATED COCKTAIL WITH ABOUT
10 GROWTH MOLECULES AND, ALLOWED
US TO TAKE A SINGLE HEPATITIS
EAT O CITE AND CONVINCE THESE TO
START DIVIDING.
SO THEY'RE SLOWER THAN THE GUT
ORGANOIDS BUT YOU CAN SEE THE
GROWTH WELL.
THIS IS MOUSE MARKED IN ALBUT
MEN EXPRESSION, WE THEN COULD
ALSO DO SOMETHING SIMILAR FOR
HUMAN LIVER WHERE THE YOUNGER
THE DONOR OF THIS WORKS AND THE
USE NOW ROUTINELY FETAL HUMAN
LIVER PROBABLY A LOT HARDER TO
WORK WITH IN THE U.S. BUT THEY
LOOK AT WHAT WE GROW, LOOKS
SIMILAR TO,A DULT LIVERS, THE
ONLY THING THEY ACTUALLY SEEM TO
BE GROW FOREVER AS ADULT LIVER
STOP AFTER 4, 6, 7, 8 MONTHS AND
WE APPRECIATE THESE LARGE CELLS,
IN THERE BUT LARGE NUCLEI, AND
BIG LIST, THE HNF 4 ALPHA
TRANSCRIPTION FACTOR MARKING
WITH THE HEPATOCYTES CLEARLY
EXPRESSED, YOU EVEN SEE A
DIVIDING HEPATOCYTE HERE BUT THE
PROTEIN WE STAYED FOR, COULD BE
ALBUMIN BUT THIS IS THE
TRANSCRIPTION ABUNKED AN
EXPRESSION, YOU DO SINGLE CELL
SEQUENCING, THESE CELLS ARE
INDISTINGUISHABLE FROM REAL
HEPATOCYTES FROM THE 1
DIFFERENCE YOU SEE A LOT OF CELL
CYCLE MARKERS EXPRESSED IN MANY
OF THE CELLS BECAUSE THEY'RE
PROLIFERATIVE.
EVEN BETTER THEY CREATE A SUPER
CELLULAR STRUCTURES AND MAYBE
THE NEXT SLIDE IS BETTER, YEAH,
HERE YOU SEE AGAIN, THESE LARGE
HEPATOCYTES, THESE THIS IS NOW
STAINED FOR A BILE DUCT, BIOACID
TRANSPORTER AND NRP2 AND THIS IS
EXPRESSED IN BILE
[INDISCERNIBLE] SMALL CHANNELS
THAT ARE FORMED BAYOUS FUSING
THE APICKAL DOMAINS OF THE 2
NEIGHBORS HEPATITIS EAT O CITES
TOGETHER AND THIS CREATING A
LITTLE CHAN AND HE WILL THE
HEPATOCYTES WILL SECRETE
BIOASSAYS THEY MAKE IN THESE
CULTURES, VERY ABUNDANTLY AND
THEY FEED INTO A CENTRAL SPACE,
WHICH NORMALLY WE THINK WOULD BE
DISPLACED BY ANOTHER BILE DUCT
TO CONNECT UP CREATED BY THE
OTHER CELL HEPATITIS EAT O CITES
TO TAKE IT OUT OF THE LIVER AND
YOU SEE THESE PATIENTS SWELL UP
AND BURST AND THAT'S HOW THEY
RELEASE THEIR BIOASSAY, AND YOU
MIGHT ALSO SEE EVERY HEPATITIS
EAT O CITE HAS MANAGED TO CREATE
A [INDISCERNIBLE] WITH ITS
NEIGHBOR.
WE HAVE NICE MOVIES WHERE WE SEE
HOW THIS PROCESS PROGRESS.
YOU CAN TRANSPLANT THESE AGAIN,
LIKE WHAT I SHOWED EARLIER FOR
THE GUT HERE AND THIS WAS
NOTHING BUT YOU COULD GO ON
NEW YORK AND MY LAB, AND THE
ORGANOIDS, THIS IS HUMAN
ALBUMIN, STAINING A LARGE
TRANSPLANTED HUMAN HEPATITIS EAT
O CITE, ORGANOIDS AND YOU SEE
THESE STAINING, THIS IS ABOUT 23
MONTHS AFTER TRANSPLANTATION,
CAN YOU SEE THE HUMANOID IS
STILL GROWING AND THIS IS DONE
IN THE BACKGROUND FOR THE MUSEUM
STANT MICE WHICH NORMAL CHAO,
SLOWLY LOSE ITS OWN HEPATITIS
EAT O CITE AND ACREATES SPACE
AND ALLOWS THESE TO GROW AND
SLOWLY FILL UP THE ENTIRE MOUSE
LIVER, AGAIN, SHOWING THAT WHAT
WE ARE GROWING HERE FROM THE
VERY FEW WITH MAYBE PROBABLY
ONLY 1 CELL, REMAINS ENTIRELY
NORMAL DESPITE THE PACT THAT WE
CAN EXPAND VERY DRAMENTIC
LOAMACYY OVERTIME.
CURRENTLY THIS IS UNPUBLISHED
BUT WE ARE TRYING TO NOW SEE IF
WE CAN USE THESE ORGANOIDS FOR
VARIOUS DISEASE MODELING HERE.
WE MIMIC CHRONIC HBV, HEPATITIS
B INFECTION WHICH IS HUGE
CHALLENGE IN AFRICA AND ASIA AND
CHINA.
AND BECAUSE HEPATOCYTES CAN NOT
BE MAINTAINED IN CULTURE, DAYS,
MAYBE WEEK, YOU CAN NOT STUDY
CHRONIC INFECTIONS SO WE PLAYED
THEM OUT IN 2 D AFTER WE GROW
THEM AND KEEP THEM IN 2 D, AND
[INDISCERNIBLE] 1 CHINESE POST
DOC CURRENTLY IN THE LAB, HE IS
MANAGED TO MAINTAIN HPV
INFECTION FOR 2 MONTHS IN THESE
CULTURES AND FOR THE INSIDE, THE
CCDNA, SORT OF LATENT FORM OF
THE HEPATITIS B VIRUS, GENOME,
IS ABUNDANTLY PRESENT IN
THESE--IN THESE CELLS AND KEEPS
ON CRANKING OUT THE HBS ANTIGEN
SO CAN YOU MEASURE IT OUTSIDE TO
CELLS BUT AS YOU SEE ALSO
VIRAL--INFECTIOUS VIRAL PARTING
ARE PRODUCED IN THESE CULTURES
SO WE THINK THIS MIGHT BE A NICE
PLATFORM TO STUDY CHRONIC HPV
INFECTIONS AND POSSIBLY DEVELOP
THERAPYINGS FOR THESE CHRONIC
HPV INFECTIONS AND ANOTHER MODEL
WE CURRENTLY, JUST SUBMIT AID
PAPER BUT THE NAME HERE, WHO'S
BEEN TRYING TO MODEL
[INDISCERNIBLE] THE FIRST STAGE
OF NONALCOHOLIC FATTY LIVER
DISEASE, SEE THE NAMES HERE AND
1 WAY OF CREATING THESE MODELS
IS BY KNOCKING OUT GENES THAT
ARE AT THE END OF THE LIPID
SECRETION PATHWAY, TO APO-B AS A
CENTRAL ELEMENT OF THE LDL
PARTICLES AND THE HEPATOCYTES
NEEDED TO SECRETE DE NOVO
SYNTHESIZED LIPITS IF YOU KNOCK
IT OUT, THERE'S ALSO HER HER
EDUCATIONALITARY SYNDROME,
THERE'S LARGE DROPLETS OF LIPIDS
SYNTHESIZED BY THE HEPATOCYTES
BUT THEY'RE NOT ABLE TO SECRETE
IT.
SO SURPRISINGLY EVEN DESPITE THE
SO HEAVILY LIPID, IT WILL
CONTINUE TO GROW, BUT I DON'T
SHOW HERE WE CAN DO THE SAME
THING WITH WILD TYPE ORGANOIDS
IF YOU GROW THEM IN HIGH
CONCENTRATIONS OF FATTY ACIDS,
THEY ALSO SHOWED THE SAME LIPID
DROPLETS, THERE'S NO REGISTERED
DRUGS AT THE MOMENT EVEN THOUGH
IT'S ESTIMATED ABOUT 20% OF ALL
ADULT AMERICANS HAVE A FATTY
LIVER WHICH MIGHT DEVELOP INTO
CIRRHOSIS AND FIBROSIS AND
EVENTUALLY INTO LIVER CANCER.
THERE'S A LARGE NUMBER OF
COMPOUNDS IN DEVELOPMENT.
WE TESTED A WHOLE SERIES OF THEM
AND WE SEE A FEW THAT ACTUALLY
IN A MATTER OF 1 OR 2 DAYS WILL
TOTALLY LEAD TO THE REMOVAL OF
THESE LIPID PARTICLES SO
BASICALLY RESTORE IN A NORMAL
SITUATION, SO WE THINK THIS
WOULD BE A NICE MODEL TO STUDY
HOW HEPATOCYTES, OR LIPIDS WHAT
YOU CAN DO ABOUT THIS.
>> OTHER HAPPENLICATIONS THIS IS
INFECTIOUS DISEASES, I JUST
SHOWED BECAUSE THE ANIMATION IS
THERE, THAT THE ALBUMIN MAKES
AND THEN THE ANIMATION IS VERY
NICE, BUT HERE WE WORK WITH ROB
O'CONNOR IN BOSTON AND TRY TO
SEE IF WE COULD GROW MICROBES
THAT COULD NOT BE GROWN IN THE
LAB ON NORMAL CELL LINES AND 1
OF THEM IS SPOREDUCATIONALIUM,
AND HERE YOU SEE THE
EXPERIMENTS, SO WHAT WE WANTED
TO DO IS ASSUMING THAT IT'S A
GUT INFECTION THAT THEY CAUSE
CHRONIC GUT INFECTIONS
PARTICULARLY IN THE THIRD WORLD
COUNTRIES SO WE ARE GOING TO
REJECT SO CALLED OCYST WHICH IS
THE INFECTIOUS STAGE LIKE AN EGG
WITH SPOROZITE INSIDE AND HOPING
THAT IN OUR ORGANIZATIONSONNOID
WE HAVE THE CELL TYPE THAT WOULD
BE REFERRED SITE FOR THESE
PARASITES TO PROLIFERATE.
YOU SEE THE OSIS LANDING INSIDE
A HUMAN SMALL INTESTINAL
ORGANIZATIONSONNOID, MOST OF
THEM IN CELL TYPE IN NEEZ
ORGANIZATIONSONNOIDS IN THESE
SIGHTS, THESE ARE THE CELLS ON
THE VILLI THAT ABSORB THIS
ACTIVITY OF THE GUT.
YOU SEE THIS COMING OUT OF THE
OCYST, WE FIND THIS IN THE REAL
EXPERIMENTS THAT THEY PREFER THE
ENTHUSIASMER O SIGHTS, SO THE
ABUNDANT CELL TYPE, YOU SEE THE
MICROVILLI HERE AND ONCE THEY DO
THIS THEY TALK ABOUT INSIDE THE
MIRON AND THEY MULTIPLY TO ABOUT
8 OF THESE, SO ESSENTIALLY, THIS
STEP, AFTER ABOUT A DAY OR SO,
THIS STAGE COMES OUT OF THE
STRUCTURE AND NOW EFFECTS A
SECONDARY ENTHUSIASMER O CITE,
SIMILARLY AGAIN FORMS A MIRO, SO
IT'S TYPE 2 MERONT, SO IT'S
READY TO GO INTO SEXUAL STAGES
ABOUT 2 DAYS INTO THE INFECTION
CYCLE.
NOW WE GET 2 WITH TYPES OF
STRUCTURES IN FRONT, WE SEE A
MIL STRUCTURE, WE SAY A FEMALE
STRUCTURE AND THEY THE MALE
STRUCTURE HAS THE MICROGAM EATS,
THE MICROGAMMONS, LIKE THE SPERM
EQUIVALENCE, WHEN THEY LEAVE THE
MICROGAMMON THEY CAN FERTILIZE,
THIS IS THE FERTILIZE THE
MACROGAMMONS AND NOW WE'RE
ACTUALLY BACK AT STAGE 1 WHERE
AN OCYST IS FORMED AND IT'S THEN
RELEASED AND NOW CAN LEAVE THE
BODY OF THE INFECTED ANIMAL, CAN
BE [INDISCERNIBLE] FOR HUMAN
BEINGS AND THEN TO BE AFFECTING
THE NEXT PARASITE.
NOW IT WORKS VERY WELL, AS CAN
YOU SEE HERE BY EM, I WON'T
SPEND TOO MUCH TIME ON THIS BUT
INDEED, THE REASON PROBABLY THAT
THIS PARAICIDE COULD NEVER BE
GROWN IN THE LAB WAS BECAUSE,
CANCER CELL LINES, AT LEAST THAT
WE KNOW ARE TRANSFORMED CELLS,
YOU DO NOT HAVE THESE FULLY
DIFFERENTIATED [INDISCERNIBLE]
AND THEREFORE THEY CANNOT
PROPAGATE THIS PARTICULAR
MICROBE BUT THEY ASKED US
EARLIER SHOWED THAT THE SAME
THING FOR NORO VIRUS CHRKS IS A
VIRUS THAT COULD NEVER BE
CULTURE INDEED THE LAB.
IT ALSO PREFERS ENTHUSIASMER O
SIGHTS AND ACTUALLY, WE TRIED,
THIS NEVER WORKS FOR US BUT MARY
REASONED THAT POSSIBLY BIOASSETS
COULD BE IMPORTANT AND THAT WAS
THE KEY TO GET THIS WORKING SO
SHE COULD SHOW THAT YOU COULD
PROPAGATE NORO-VIRUS IN THE LAB
USING THE ORGANOIDS WITH THE
DIFFERENTIATED CELLS.
I THINK IT WAS VERY PRODUCTIVE
TO USE THIS TECHNOLOGY FOR
CANCER RESEARCH, WE AND MANY
OTHER LABS PUBLISHED EXTENSIVELY
ON GENERATING ORGANOIDS FROM ALL
SORTS OF CARCINOMAS, GUT, LUNG,
PROSTATE, BREAST, TYPICALLY,
WHEN YOU USE THE BASIC PROCESSES
PROT COL, SOMETHING WILL GROW
AND IMPROVE.
ALSO SIDE BY SIDE, CAN YOU GROW
HEALTHY TISSUE AND CANCER TISSUE
FROM THE SAME PATIENT, WITH
CRISPR, CAN YOU TAKE HEALTHY
TISSUE AND TURN IT INTO 4 OR 5
MUTATIONS FROM CANCER FROM
CRISPR, CAN YOU REMOVE THEM AND
SEE WHAT THAT MUTATION
CONTRIBUTED TO AS THE PHENOTYPE.
SO THAT'S ALL DESCRIBED, YOU CAN
USE IT FOR PERSONALIZED MEDICINE
AND LARGE NUMBER OF STUDIES NOW
SORT OF VALIDATING ORGANOIDS AS
AN AVATAR OF PATIENTS SO TEST
DRUGS ON ORGANIZATIONSONNOIDS
MIGHT PREDICT WHAT WOULD BE GOOD
DRUG COMBINATION TO TREAT
PATIENTS.
THIS IS A RECENT STUDY, VERY
DIFFERENT FROM WHAT I JUST
DESCRIBED, PUBLISHED LAST YEAR
AND ACTUALLY THESE 3 PIECE,
[INDISCERNIBLE] AND MY LAB AND
[INDISCERNIBLE] RUBEN'S LAB CAME
UP WITH THIS EXPERIMENT AND THEY
SHOWED THE RESULTS BEFORE RUBEN
OR I RENEWED THIS PROJECT.
THEY STARTED READING ABOUT A
VERSION OF E.COLI, SO A VERY
COMMON MICROBIOME COMPONENT AND
THESE PARTICULAR E.COLI HAVE AN
EXTRA BIT OF DNA ABOUT 60 DILL O
BASES CALLED POLYKETIDE
SYNTHESIS, PKAS POSITIVE,
CONTAINING 10 GENES MOSTLY
ENZYMES TOGETHER CONITUTE A
SYNTHESIS PATHWAY, AND THIS
PARTICULAR POLYKETO VECTOR
GENOMEIN, THESE HAVE BEEN KNOWN
FOR ABOUT 30 YEARS TO BE
PHONOTOXIC SO WHEN YOU CULTURE
THE EON COAL I WITH THAT EXTRA
PENE OF THE CHROMOSOME, YOU CALL
TO THEM ON TOP OF HUMAN CELL
LINES YOU RAPIDLY SEE DOUBLE
STRAND BREAKS THAT,'S WHY
THEY'RE CALLED GENO TOXIC AND
THE QUESTION THAT THE 3
PHYSICIANS ASKED IS ESSENTIALLY
CAN SECRETE THIS AND EXPOSURE TO
THIS LEAD TO MUTATIONS AND THE
PARTICULAR CAN LEAD TO CANCER
MUTATIONS, SO NOT ONLY CAUSING
DOUBLE STRAND BREAKS AND ALSO IN
THE RESOLUTION OF THESE DOUBLE
STRAND BREAKS RESULT IN
MUTATIONS THAT WOULD THEN
ULTIMATELY LEAD TO CANCER, BUT
THIS IS COLI BACTERIAIN, AND
WHILE WE WERE DOING THIS IN THIS
PAPER ARE BIND TO SPECIFICALLY
TO A-RESIDUES AND DNA AND
MODELED HERE, CROSS LINK THIS IS
NOT COMPATIBLE WITH THE LIFE OF
THE CELL APPARENTLY IN THIS
PROCESS, THE CELL CAN UNDERGO
DOUBLE TRAND BREAKS AND THAT
WOULD MOST LIKELY LEAD TO THE
DEATH OF THE CELL, SO THAT'S WHY
IN THE RESOLUTION OF DOING THE
E.COLI, THE MUTANT, DERIVED FROM
THIS, ON THE LEFT, WE SEE IT ON
THE RIGHT, WHICH LACKS 1 OF
THESE GENES AND SO THIS IS THE
BAD GUY ON THE RIGHT, HAVE YOU
THE CONTROL FOR THAT GUY, AND
THE OTHER WOULD BE THAT NOW WE
GROW THEM ON OUTER PLATES WE ARE
NOT MICROBIOLOGIST BUT THAT'S
EASY, WE THEN PICKED THEM UP AND
THEY HAVE GLASS NEEDLES WE USE
THESE INJECTORS FOR THIS AND
THEN WE WOULD INJECT INDIVIDUAL
ORGANOIDS WITH THESE BACTERIA,
LEAVE THEM IN FOR 1 OR 2 DAYS
AND THEN ASK AND SEE WHAT WAS
PUBLISHED PREVIOUSLY IN THE
DOUBLE STRAND BREAKS.
AND THEN OF COURSE ON THE LEFT
WE SEE THEM AND ON THE RIGHT
THERE WOULD BE NEGATIVE CONTROL.
AND ON THE LEFT ABOUT MAYBE 40
OR 50 ORGANIZATIONSONNOIDS, 1 BY
1 INJECTED BY [INDISCERNIBLE]
WITH THE PKS BACTERIA AND INDEED
ON THE GLOW, 1 OF THESE SEE MANY
OF THESE RED DOTS, WHICH
ESSENTIALLY ARE GAMMA H2 X FOCI
WHICH REVEAL THE PRESENCE OF A
DOUBLE STRAND BREAK.
SO INDEED IN THE CONTROL WE
DON'T SEE IT, SO WE ASK,
YES,--IT WILL OVERGROW THE
CULTURE AND THE END OF THE
EXPERIMENT SO IT IS FOR ABOUT 3
MONTHS AND REALLY MIMICKING
CHRONIC EXPOSURE TO PKS E.COLI
THAN THE CONTROLS ON THE RIGHT
YOU WOULD PROBABLY STILL HAVE
THE HATTER TIME WITH THE
MUTATIONS BECAUSE EVERY CELL
MIGHT HAVE A DIFFERENT
COMPLEMENT OF THESE SMALL
LESIONS AND INSERTIONS, SO THEN
WE DID THE SUBCLONES AFTER 3
MONTH EXPOSURE, THESE CELLS GREW
THEM UP, ESSENTIALLY TO AMPRIFY
THE GENOMES OF THESE SINGLE
CELLS, TO AN AMOUNT WHERE WE
COULD SEEQUENCE THESE ORGANOIDS
AND IN ESSENCE READ OUT THE
GENOME OF THE CELL THATULENT TO
THE SUBCOLON, AND THIS ALLOWED
US TO COUNT MUTATIONS AND
INDEEDOT LEFT, THE MUTATIONS
THAN ARE ON THE RIGHT BUT MORE
IMPORTANTLY AND THIS WAS USING
MIKE STRATEGIC PLANON'S WAY OF
USING DNA SIGNATURES OR LOOKING
FOR WHAT KINDS OF MUTATIONS YOU
CAN INDUCE AND I WILL SHOW THIS
BETTER IN THE NEXT SLIDE, BUT WE
FOUND OUT THAT INDEED THIS WOULD
BE THE NORMAL MUTATIONS YOU SEE
IN CELL CULTURE, SO CELLS IN
CULTURE SUSTAIN RANDOM MUTATIONS
OF SOME LEVEL, ASK PARTICULARLY
WHEN YOU SCORE FOR T TO C OR
CHANGES OR T-G, AND WE SEE PEEKS
WE MED NOT SEE IN THE CONTROLS.
IMH WE VERY CAREFULLY MET THE
SINGLE BASE CHANGES OR DELETIONS
IN THE EXPOSED ORGANOIDS AND
MAKE A STRIKING DISCOVERY.
SO THIS GOES FOR A VERY UNIQUE
MOTIF, ALMOST LIKE A RESTRICTION
ENZYME OR A TRANSCRIPTION
FACTOR, SO YOU SEE, A T 8 THERE
IS EITHER DELETED OR MUTATED BUT
IS DONE IN A CONTEXT OF AT RICH
DOMAIN AND A ON THE MINUS 3
POSITION.
AND IF YOU MODEL THIS AGAIN THE
WAY I SHOWED YOU EARLIER, WE
BELIEVE WHAT'S HAPPENING IS THAT
THIS BINDS TO THE POSITIVE
STRAND SO THIS WOULD BE THIS A
AND THIS BINDS COVALENTLY TO AN
A ON THE NEGATIVE STRAND, AND
THE OPPOSITE THE T OF HERE, AND
NOW THE CELL AS A PROBLEM, IT
HAS TO RID OF THIS ADDUCT AND
WHILE IT GETS RID OF IT, IT
APPARENTLY CHANGES HERE INTO ANY
OTHER BASE OR IT DELETES THIS,
NOW IF THIS HAPPENS IN A READING
FRAME, CHANGING IT IN ANY OTHER
BASE COULD CAUSE A MISSED SENSE
MUTATION AND DELETING THIS
SINGLE BASE PAIR WOULD LEAD TO A
FRAME SHIFT IN BOTH CASES OF
COURSE CAN CONTRIBUTE TO CANCER.
NOW THESE MUTATIONS OCCUR
RARELY, BUT IN THIS ASSIGNMENT
WE SEQUENCED THE ENTIRE GENOME
AND WE SEE WHERE IT HITS THE
GENOME AND WE DON'T THINK IT HAS
PREFERENCE FOR A GENE TJUST GOES
FOR AT RICH SEQUENCES AND IT
WILL CHANGEERER REMOVE THIS
THERE.
SO VERY ARTIFICIAL, WE THEN
ASKED DOES IT HAPPEN IN REAL
LIFE THEY WERE VERY UNUSUAL,
THEY HAVE NEVER BEEN SEEN BEFORE
WE WERE LUCKY TO HAVE ACCESS TO
THE MEDICAL FOUNDATION
COLLECTION AND SO THEY'VE BEEN
SEQUENCING WHOLE GENOME,
METASTASIS FOR A LARGE VARIETY
OF DIFFERENT HUMAN CANCERS WE
ASK WHICH OF THESE CANCERS DO WE
SEE SIGNS OF THIS PARTICULAR
MUTATIONAL PROCESS, AND LOW AND
BEHOLD WE DO SEE THAT MAYBE WE
DO THINK ABOUT 15% OF ALL COLON
CANCERS SHOW CLEAR SIGNS OF
THESE MUTATIONS SO THIS SCORES
HOW MANY MUTATIONS FIT WHAT I
JUST SHOWED YOU FOR THE
SIGNATURE, BY COMPARISON, THERE
ARE QUITE A FEW BREAST CANCER
CASES IN THIS COLLECTION, THERE
WAS NOT A SINGLE BREAST CANCER
CASE WHERE WE SAW ANY EVIDENCE
OF THIS TYPE OF MUTATIONS, YOU
GET THE ON OCCASIONAL HEAD AND
NECK CANCER WE'VE SEEN A FEW
MORE, OCCASIONALLY BLADDER
CANCERS HERE, AND THOSE ARE
SIGNS WHERE E.COLI CAN LIVE, SO
POSSIBLY THESE ARE ALSO CAUSED
BY PKS E.COLI.
AND THEN IN THE SECOND COHORT,
THIS IS COLORECTAL CANCERS,
AGAIN, THE SAME PREVALENCE WAS
FOUND WITH MAYBE 5 TO 10% OF THE
COLON CANCER PATIENTS HAD THIS
PARTICULAR SIGNATURE AND HAVE
IMPROVED ALGORITHM AND THE
RIBBON, WHOSE LAB IS PURSUING
BIO-INFORMATICS OF THIS,
BELIEVES THAT 15% OF COLOON
CANCERS HAVE LARGE NUMBERS OR
FEWER NUMBERS WITH CLEAR
EVIDENCE OF THIS MUTATIONAL
PROCESS.
WE THEN ASKED, SO IT'S--SO FAR
BEFORE WE LOOKED AT DO WE HIT
CANCER GENES, NO, THE KRASE
MODEL ARE QUITE COLON IN COLON
CANCER, THIS CANNOT BE CREATED
BY THIS MUTATIONAL PROCESS
BECAUSE THE MUTATIONAL FOOTPRINT
IS NOT THERE, THE RICH SEQUENCE,
APC, WHICH IS THE MOST COMMON
TUMOR SUPPRESSOR IN COLON CANCER
MUTATED UP TO 90% OF OF THE
RELIEF TO CARRY OUT AND THESE
CANCER CASES WE FIND.
THE WITH THE APC, SO E.COLI IS
REDUCING MUTATIONS AND SEE
SPECIFICALLY KNOW FROM
LITERATURE THAT ABOUT 10% OF THE
POPULATION CARRIES BACTERIA AT
ANY GIVEN TIME, OF COURSE
BACTERIA DON'T HAVE TO BE
PRESENT IN E.COLI CANCER
PATIENT, WHEN CANCER EMERGES, IT
MIGHT HAVE HAPPENED MUCH EARLIER
IN LIFE, BUT BASED ON THIS, IT
IS CONFIRMED, COLO VECTOR IS THE
ECOLI THAT PROVES CAUSING CANCER
DIRECTLY IN UH ATHLETICS HANS
AND THIS MIGHT EVENTUALLY LEAD
TO MAYBE FIND OUT WHO CARRIES
THESE BACTERIA TO QUITE EASILY
REMOVE THEM WITH ANTIBIOTICS AND
THAT COULD LEAD TO A SITUATION
WHERE YOU PREVENT COLAN CANCER
PARTICULARLY WE THINK ABOUT HIGH
RISK PATIENTS LIKE HER
EDUCATIONALITARY COLON CANCER,
PATIENTS OR PATIENTS WITH
CHRONIC INFLAMMATORY DISEASE
WHERE YOU DON'T WANT ANOTHER
EVENT AND THEN A FINAL STORY, I
THINK I HAVE TIME FOR THIS, OR
WHEN COVID BROKE OUT 2 YEARS
AGO, WE, LIKE MANY COUNTRIES
WENT INTO LOCK DOWN AND WE ARE
ONLY ALLOWED TO DO THE MOST
ESSENTIAL EXPERIMENTS BUT WE ARE
ENCOURAGED TO DO COVID
EXPERIMENTS SO WE TEAMED UP WITH
A CORONA LAB IN ROTTER DAM LED
BY [INDISCERNIBLE],
COLLABORATORS THERE, AND THE
QUESTION WAS QUITE SIMPLE THAT
WE ASKED, SO WE ASK, IS THE GUT
AND THIS IS MARCH 2020, SO IS
THE GUT ALSO A TARGET FOR THIS
VIRUS, AND THEN WE KNOW A FEW
THINGS THEN, ACE 2, TO BE
XREAMLY HIGHLY EXPRESSED IN THE
GUT, MUCH HIGHER THAN IN THE AIR
WAYS AND ACE 2 WAS DONE ALREADY
AND RECOGNIZES RECEPTOR FOR
SARS-COV-2, AND WAS RAPIDLY
FOUND BECAUSE THERE WAS ALSO A
RECEPTOR FOR THE ORIGINAL SARS
VIRUS 20 YEARS AGO, WE ALSO
NOTED THAT QUITE SIGNIFICANT
PERCENTAGE OF COVID-19 PATIENTS,
SHOW UP WITH THE GI SYMPTOMS, SO
NAUSEA, TOMAC ACHES AND SOME THE
FIRST PC R TESTS WERE BECOMING
AVAILABLE BASED ON THE GENOME OF
THE VIRUS, AND PEOPLE HAD SHOWN
THE VIRAL RNA DETECTABLE IN
STOOL, SO IT CAME OUT OF THE
LUNGS AND ENDED UP IN STOOL
POSSIBLY IT WAS PRODUCED BY
INFECTED CELLS IN THE GUT, SO WE
ASKED VERY SIMPLY EVERY ORGANOID
CAN BE IN FACT ORGANOIDS, WORKED
BUT THE GOOD ORGANOIDS ARE MUCH
EASIER TO INFECT AND THEY
PRODUCE MUCH LARGER VIRAL
NUMBERS, VIRAL PARTICLES UV
SCORE FOR A LIFE VIRUS AND ONLY
2 AND HALF DAYS WE GET AN
INCREASE IN VIRAL PARTICLES,
QUITE SURPRISINGLY THESE CELLS
KEEP ON LOOKING FOR HEALTHY,
THEY KEEP ON GROWING FOR RNA
SAME THING AND THEY ONLY HAVE
AND PORNLLY EXPLAINING WHY
INDIVIDUALS COULD PRODUCE SO
MUCH VIRUS IN THE BEGINNING OF
THEIR DISEASE AND THAT WAY BEING
FANTASTIC SPREADERS OF THE VIRAL
INFECTIONS, SO YOU SEE A SINGLE
INFECTED CELL BY EN, WE SULLY
SEE THAT THE RIEROUS WILL ATTACH
TO THE APEX OF THE CELLS SO THIS
IS WHERE THE ACE 2 SITS AND SO
THE INFECTION COME FROM THE
LUMEN, MUCH LIKE THEY ENTER THE
PATIENT, NO INTESTINAL FACTOR IN
THE LUNGS, THAT'S HERE, SITS ON
THE BASAL SIDE, STAIN FOR
NUCLEAR PROTEIN HERE AND WIDE,
YOU ALSO HAVE THE VIRUS IS
SECRETED AGAIN TO THE LUMINAL
SIDE OF THE CELL AND INDEED
AFTER 60 HOURS YOU CAN SEE THAT
IN THE INFECTED ORGANOID, MANY
CELLS GET INFECTED BUT
NEIGHBORING ORGANOIDS ARE NOT
PRIMARILY INFECTED DON'T APPEAR
TO BE SENSITIVE TO THE VIRUS, SO
THEY HAVE TO ENTER THROUGH THE
APEX AND LEADS THE CELL AGAIN SO
THE ACHE PICKA MEMBRANE AND THEN
BECOMES A LITTLE BIT IMPORTANT
IN THE LAST BIT OF THE TALK.
SO IT WAS PUBLISHED RAPIDLY AND
REVIEWED OVERNIGHT THE JOURNAL
IS PUBLISHED OVERNIGHT AND
ACTUALLY SEVERAL OTHER PAPERS
SHOWING SOMETHING SIMILAR AND
VERY NICE NATURE MEDICINE FROM
JANE IN HONG KONG, SHE SHOWED
YOU CAN QUITE EASILY GROW BAT
ORGANOIDS FROM THE GUTS OF BATS
AND THEY ARE ACCUSTOMED TO MANY
CORONAVIRUSS IN HUMANS SO IT'S
SEQUENCES THAT ARE NEVER GROWN,
THEY COME FROM BATS AND WE
PROPOSE THESE BATS ORGANOIDS
WILL BE GREAT TO TBROA THESE
CORONAVIRUSS, BUT YES,
REGULATORY AUTHORITIES ARE VERY
MUCH AFRAID OF GREEN CELLING
ADDITIONAL SPECIES OF COROWN A
VIRUS IN THE LAB SO I DON'T
THINK THAT HAS BEEN FOLLOWED BY
[INDISCERNIBLE].
THIS STUDY HAS NOT BEEN
PUBLISHED BUT HERE WE ASK WHAT
ARE THE HOST GENES THAT ARE USED
BY THESE VIRUSES?
AND THERE WERE THEN ALREADY A
FEW PAPERS MOSTLY IN CELL WHERE
GENOME WIDE CRSPR HAD BEEN DONE,
THE LINES THAT HAD BEEN DONE,
AND MAYBE 50, 60 Gs HAVE BEEN
PROPOSED FOR THE VIRAL
INFECTIOUS CYCLE, ACE 2 I
MENTIONED IT, THE RECEPTOR THIS
WAS A CONTROL HERE SO WHAT WE DO
HERE AND WE DON'T DO BAR CODE,
TRANSFECTIONS WE BASICALLY GO
CLONE BY CLONE AND CANDIDATE
GENES SO THIS IS WITH A NUMBER
OF MUTANTS WE SEQUENCE THEM AND
CONFIRM THEY ARE NULLS AND WE
USE THEM AND THEN WIEWS WE USE
THEM AND AS YOU CAN SEE HERE,
COMPARED TO WILD TYPE, THEY CAN
NOT BE EFFECTED BY SARS 2, AND
THEY CANNOT BE INFECTED BY THE
ORIGINAL SARS VIRUS ABOUT THEY
ARE INFECTED BY A THIRD SPECIES
HERE RELATED TO SPECIES FIRST
AND CORONAVIRUS HAS BEEN AROUND
IN OTHER COUNTRIES AND APPEARS
TO BE JUMPING FROM CAMELS AND
HUMANS AND VERY FEW REPORT OF
THIS VIRUS JUMPS FROM HUMANS TO
HUMANS.
ONE THING WITH SARS-COV-2 IS
PEOPLE GET VERY SICK RIGHT AWAY
AND HIGH LEATH ALGORITHMSITY,
50% OR SO, BUT WE ALSO SEE THAT
THE CULTURE IS ESSENTIALLY THAT
IN 2 OR 3 DAYS, SO THE MOMENT
THE VIRUS STARTS PROPAGATING
ITSELF, IT KILLS THE CELLS AND
THAT'S MAYBE WHY MRS HAS NOT
SPREAD SO FAR COMPARED TO
SARS-COV-2.
SO THAT'S POSITIVE CONTROL.
ANOTHER CONTROL HERE IS KNOWN TO
BE THE RECEPTOR OF MERS, HAS
BEEN PROPOSED TO BE A RECEPTOR
FOR SARS-COV-2, HOWEVER WHEN WE
KNOCK OUT DP 4, THIS AFFECTS
HERE IN RED COMPARED TO GREEN
THE CONTROL, BUT NOW MERS, VIRUS
IS CONTROL AND THESE ARE PP4
CONTROL MUTANTS SO THIS CONNIRMS
CAN YOU READ OUT IN THESE MUTANT
ORGANOIDS WHAT THE VIRUS NEEDS
TO PROPAGATE AND STRESS THIS
PRIMARYY EPITHELIUM AND FIBRO
BLAST LIKE CELL LINE AND WE THEN
WENT THROUGH A LONG LIST OF
THESE PUBLISHED GENES AND MUCH
TO OUR SURPRISE NONE OF THESE
GAVE US A PHENOTYPE SO WE COULD
KNOCK OUT QUITE A FEW WITHOUT
ANY EFFECT OF A SINGLE GENE,
REALLY WELL STUDIED, TEMPRESS 2
IS A PROTEASE DOMAIN ON THE
OUTSIDE ON THE APICKAL SIDE OF
THE CELL, THIS IS KNOWN TO
CLEAVE THE SPIKE PROTEIN OF THE
VIRUS AND THEN ALLOWS FUSION OF
THE VIRAL MEMBRANE AND VIRAL
ENVELOPE WITH THE HOST CELL.
SO VIRUS KNOCK OUT, IT CANNOT
EFFECT, THIS IS THE RULE LINES
HERE, KNOCKED OUT OF THE
EXPRESSION OF EPITHELIAL
TISSUES, THEY HAVE NO EFFECT SO
WE WOULD CONCLUDE THAT TMPRSS2
WOULD BE A GREAT DRUG TARGET.
TMPRSS2 ARE 9 AS PUBLISHED SO IT
SHOULD BE SAFE AND JUST LAST
WEEK I THINK IS A PARTNERSHIP
THAT APPEAR INDEED BIOARCHIVES
WITH AN INHIBITOR OF THE TMPRS
PROTEASE BUT IT WILL NOT MUTATE
WHERE THE VIRUS STARTS TOGETHER
AROUND THE DRUG AND IT WAS SHOWN
TO BE EFFECTIVE BLOCKER OF VIRAL
INFECTION AND THIS WILL NOT ONLY
WORK FOR SARS-COV-2 BUT
HOPEFULLY IT WILL WORK FOR MANY
OTHER CORONAVIRUSS AND WE THINK
IT'S A FANTASTIC DRUG TARGET.
NONE OF THESE OTHERS REALLY
WORKED I DRAW YOUR TENSION TO
CTSL HERE IN PURPLE HERE, IT'S
THE PATHWAY THAT CAME UP IN MOST
OF THE SCREENS AND IT ENHANCES
ACTIVITY BILET VIRUS SO IT
DOESN'T BLOCK IT AT ALL.
QUITE SURPRISING AND THEN WE
REPEATED EXPERIMENTS FOR
LITERATURE, CHLOR O QUINN,
HYDROXYCHLOROQUINE WHEN USED ON
VOE 6 CELLS, WE USE THESE CELLS
IN THE SCREENS, AT LEAST IN MOST
OF THESE AND FANTASTIC INHIBITOR
OF VIRAL REP REPLICATION AND 1
MICROMOLAR WE REACHED IT AT 10
MICROMOLAR THERE ARE NO VIRAL
PROPAGATION IN THESE VERY 6
CELLS, LET ME KNOW, TAKE 10
MICROMOLAR IN THE PRIMEAR
EPITHELIAL STRUCTURE AS
ORGANIZATIONSONNOIDS HERE IN
BLUE AND DOES NOTHING.
NOW HOW DO WE PUT ALL THIS
TOGETHER, AND INTO A SINGLE
STORY, I SHOULD STRESS AGAIN WE
AREN'T THE ONLY 1S THAT POSE
THIS OR PAPERS THAT STAIM TIME
AS WE PROPOSE THIS SO WHAT
HAPPENS IN THESE WORK HORSE CELL
LINES, AS TYPICALLY MORE FIBRO
BLAST LIKE THE WAY THEY GROW,
THEY ARE DEFINITELY NOT LIKE
EPITHELIA, AND THEY DON'T
EXPRESS ACE 2 SO YOU NEED TO
TRANSFECT ACE 2 TO MAKE THEM
SENSITIVE TO THE VIRUS, THEY CAN
THEN BIND THE ENDOSIGNIFYITOSE
AND THIS ENDOSIGNIFYITOSEIS STEP
IS ARE READILY BLOCKABLE BY
HYDROXYCHLOROQUINE, THAT'S ALSO
HOW IT WORKS ALSO IN MALARIA AND
THEN TO RELEASE THE VIRUS FROM
THE ENDOSTUDIES OF MULTIPLE
ENDOCRINE THIS IS WHERE THEY'RE
CALLED IN SO IT'S NO WONDER THAT
THIS IS LIKE CONCEPTIONS CAME UP
IN THE WIDE SCREENS, WHAT
HAPPENS IN PRIMARY EPITHELIA,
THIS WOULD BE THE ACHE PICKA
SIDE OF EPITHELIUM, ACE 2 IS
NATURALLY EXPRESSED, TMPRSS2 IS
EXPRESSED ON THE APICKA
MEMBRANE, THE VIRUS BINDS TO ACE
2, TMPRSS2 ACTIVATES BY PROTEASE
CLEAVAGE DESPITE THAT SITS ON
THE OUTSIDE OF THE CORONAVIRUS
AND NOW, THE VIRUS DIRECTLY
FUSES IT'S ENVELOPE WITH THE
HOST CELL MEMBRANE AND CAN
INJECT THE RNA MOLECULES
STRAIGHT INTO THE CYTOPLASM OF
THE CELL.
THIS DOES NOT INVOLVE
ENDOSIGNIFYITOSEIS SO THERE IS
NO SENSITIVITY TO HYDROXY LORA
RODRIGUEZ CLOA QUINN AND NO
REASON WHY TRANSLATIONAL
RESEARCH TRANSITION SHOULD BE
INVOLVED IN THIS STEP.
THAT'S WAY FIND.
THEY ARE NOT THERE.
SO FROM THIS WE WERE ABLE TO
DEDUCE THAT IT'S PROBABLY A GOOD
IDEA IF 1 DOES NO SCREENS,
HITHROUGH PUT SCREENS, IN EASY
CELL LINES, EASY TO WORK WITH,
THE HITS MAYBE WOULD BE GOOD TO
ALSO CONFIRM IN TISSUES THAT ARE
MORE CLOSE TO PRIMARY EPITHELIUM
IN THIS CASE, SUCH AS AIR WAY OR
GUT ORGANOIDS AND AGAIN, VIRUSES
THAT ENTER THE AIR WAYS OR
INTESTINAL TRACT, IT WILL ALWAYS
COME IN THROUGH THE APICKA SIDE,
SO THERE WE BELIEVE THE THESE
ARE SUITED BECAUSE CAN YOU
INJECT YOUR, WHATEVER AGENT INTO
THE LUMENS OF THE ORGANIZATIONS
KNOWOIDS IT WOULD BE HERE AND
YOU CAN GROW THEM WHEN THEY'RE
EXPAND INDEED 2 D IF YOU KEEP
THEM FOR A FEW WEEKS AND THEN
JUST ADD WHATEVER YOU WANT TO
ADD ON THE TOP SIDE OF THE
CULTURE ON THE APICKA SIDE AND
ANS WELL, FLIPPING AROUND FOR
THE BASAL SIDE OF THE EPITHELIUM
AND WITH THAT I REACH THE END OF
MY TALK.
I BELIEVE MENTIONED ALL OUR
COLLABORATORS DURING THE TALK SO
I WILL STOP SHARE AND I HOPE I
LEFT TIME FOR Q ANDA.
>> GREAT, THANKS, HANS, THAT WAS
GREAT PRESENTATION TO TALK ABOUT
ORGANOIDS TO MIMIC HUMAN TISSUE
TO STUDY HUMAN DISEASE.
VERY INTERESTING, THERE ARE A
FEW QUESTIONS AND MAYBE I WILL
TAKE THE FIRST.
IF I UNDERSTOOD CORRECTLY, I
THINK THAT YOUR CULTURE OF FETAL
VERSUS ADULT LIVER, ARE REALLY
MIMICKING THE AGING PROCESS,
SO--SO HAVE YOU AN INVITRO
[INDISCERNIBLE] THERE THAT
HOPEFULLY IS ACCURATE AND I'M
WONDER FIGURE YOU ANALYZED IT,
IS IT TELOMERE EROSION,
EPIGENETIC DIFFERENCES?
DO YOU KNOW WHAT'S GOING ON?
>> YEAH, SO, ACTUALLY WE DID A
LOT OF--ALL SORTS OF GENOMICS
BUT THE PRINCIPLE OBSERVATION
THAT THE THALAMUS WERE WORKING
AND WE DON'T SEE THAT IN MOST
OTHER CULTURE SO FOR INSTANCE
DIRT IS HIGHLY EXPRESSED IN OUR
GUT ORGANIZATIONSONNOIDS BUT IN
THE LIVER IT'S LIMITING THING,
ALSO IN LIVER CANCERS, IT'S
ACTIVATING MUTATIONS ARE QUITE
COMMON, SO IT LOOKS LIKE IN THE
LIVER, IF YOU WANT TO
PROLIFERATE, YOU NEED TO DO
SOMETHING ABOUT YOUR
[INDISCERNIBLE], WE CAN OVER
COME IT NOTEB ENTIRELY BUT BY
DERT, EXPRESSION AND BY
TRANSVEKS OF DERT, AND YOU SEE
THE TYPICAL SENESCENSEL AND THEN
P16 COME UP AND IT LOOKS LIKE
CLASSIC SEN ILLEGALSEN CESS
ENSEL WHICH WE NEVER SEE IN
OTHER, STILL WE DON'T KNOW
WHETHER IT'S LACK OF 1 GROWTH
FACTOR THAT WOULD SOLVE THIS OR
WHETHER IT'S SORT OF BAKED IN
THE LIVER.
BUT THE FETAL LIVER INDEED, BUT
MANY SAMPLES WE CAN GROW FOR
YEARS THEY LOOK IF THEY DO RNA
SEQ, THEY ARE ALMOST IDENTICAL
TO THE ADULT LIVERS, SOME
CYTOCHROMES MAY NOT BE AT THE
ADULT LEVEL, YOU PLAY AROUND
WITH GROWTH FACTOR CONDITIONS
AND THERE'S 1 BIG DIFFERENCE
WHERE THEY WILL GROW FOREVER AND
THE ADULLS DON'T.
>> YEAH, THAT'S REALLY
INTERESTING, PROSTATE ACTUALLY
SENESCS AS WELL, SO THERE ARE A
FEW QUESTIONS HERE,
[INDISCERNIBLE] HAD A QUESTION
ABOUT THE BOWEL REPAIR
EXPERIMENTS AND SHE'S WONDERING
ABOUT THE PLASTICITY OF THE STEM
CELLS IN THE GUT SO IF YOU WERE
TO USE STEM CELLS FROM
ESOPHAGUS, GASTRIC, SMALL,
LARGE, BOWEL, WHAT WOULD YOU
EXPECT?
>> THAT'S A VERY GOOD QUESTION.
ACTUALLY, SO THERE ARE MY FORMER
POST DOC WHO ESSENTIALLY
DEVELOPED ORGANIZATIONSONNOID
TECHNOLOGY IN MY LAB, IN HIS OWN
LAB HAS DONE A LOT OF
TRANSPLANTS AND WHAT HE FINDS IN
ESSENCE IS THAT SO ONCE THE
TISSUES ARE ESTABLISHED, SO IF
YOU TAKE SAMPLES LATE FETAL OR
ADULT, THEY KNOW WHAT THEY ARE
AND YOU CANNOT CHANGE THAT FATE.
SO THEY CAN ACTUALLY--STEM CELLS
CAN [INDISCERNIBLE] AND THAT I
CAN MOVE BACK AND FORTH SO
THERE'S PLASTICITY IN THE
LINEAGE BUT IF YOU TRANSPLANT A
SMALL--I SHOULD SAY, YEAH, A
SMALL INTESTINAL ORGANOID TO
COLON IT DOESN'T HAVE VILLI, IT
DOESN'T HAVE THE CELLS YOU WILL
GET THE COLON WITH VILLI AND
CELLS, THERE'S A BEAUTIFUL
NATURE RECENTLY WHERE IT SHOWS
SORT BOWEL DISEASE WHICH IS A
TERRIBLEIN SYNDROME OF INFANTS
WHEN THEY LOSE A LARGE PART OF
THE SMALL INTESTINE, THE COLON
IS FINE BAH IT DOESN'T HELP BUT
THE SMALL INTESTINE IS WHERE YOU
ABSORB THE NUCLEOTIDES T-REGENTS
AND IT IN IT CASE IT ONLY
ABSORBS WATER, SO HE CONVERTED
COLON INTO SMALL INTESTINE AND
THIS WAS DONE IN ANIMALS AND WE
HOPE TO DO THIS IN THE BABIES
AND YOU TAKE PART OF THE COLON,
STRIP THE EPITHELIUM OFF, SEED
IT WITH SMALL INTESTINAL
ORGANOIDS AND BEING BE GROWN
FROM THE PATIENT YOU ARE
TRANSPLANTING INVITRO AND
THOUGH--SO FIRST HE CREATES A
LOOP WITH 2 ENDS ON THE OUTSIDE
THROUGH THE SKIN AND THEN YOU
HAVE MANIPULATE THE COLON SO
THEN IT'S SEEDED WITH THE SMALL
INTESTINAL EPITHELIUM, HE TAKES
THAT LOOP AND PLACES IT IN THE
SMALL INTESTINAL TRACT AND THEN
IT WORKS.
>> WOW.
>> SO DEVELOPMENTAL BIOLOGY
EXPERIMENTS WHERE IT'S THE
SUBEPITHELIAL STRUCTURES, BUT
APPARENTLY LATER IN LIFE, SO
ONCE THEY HAVE THEIR FATE, YOU
CANNOT MOVE THEM.
YOU CAN NOT TURN STOMACH INTO
NAWL INTESTINE AND THE LENGTH OF
THE SMALL INTESTINES THERE ARE
REGIONS THAT ARE QUITE DIFFERENT
IN WHAT THEY TAKE UP, THEY KNOW
WHAT THEY ARE, AND EVEN AFTER
HERE IN CULTURE, THEY ARE STILL
ILIUM OR JUJUNUM, OR TRANSPLANT
THEM, THAT'S WHAT THEY MAKE.
>> WELL IN SOME WAYS THAT MAKES
THE SYSTEM, GIVES YOU CONFIDENCE
THAT YOU'RE LOOKING AT THE RIGHT
THING WHEN YOU DEVELOP THESE
ORGANOIDS TO MIMIC THE TISSUE,
IT'S VERY DIFFERENT FROM IPS
CELLS WHERE YOU PUSH THEM IN ANY
DIRECTION AND THE IPS GUT
ORGANOIDS TEND TO BE A MIX OF
THE DIFFERENT AREAS.
>> YEAH, ABSOLUTELY.
YEAH, OKAY, THERE IS A QUESTION
FROM JIM FLEET, HE SAYS, OTHERS
HAVE NOW SHOWN THAT TRADITIONAL
CULTURING OF INTESTINAL
ORGANOIDS LEADS TO MANY MORE
MUTATIONS FROMOXIDATIVE STRESS
THAN WHAT 1 SEES INVIVO.
HAVE YOU CULTURED ORGANOIDS IN A
RELATIVE HIGHWAY POXIC--IN
RELATIVE HIPOXIC ENVIRONMENT TO
MIMIC NORMAL PHYSIOLOGY.
>> THIS WAS SHOWN BY OUR
COLLABORATORS, AND SOME OTHER
LABS SHOWING THIS, THE
MUTATIONAL RATE IS 4 OR TIMES
HYPER AND INVIVO, INVIVO
ACTUALLY COULD USE THIS
TECHNOLOGY TO SHOW THAT A GUT
STEM CELL IN HUMANS SUSTAINED
ABOUT 40 SINGLE BASE CHANGES PER
YEAR OF LIFE SO 1 PER WEEK.
AND IT'S LINEAR THROUGHOUT LIFE.
LIVER THE SAME THING ALTHOUGH
THE CELLS DON'T DIVIDE MUCH,
ALSO ABOUT 1 PER WEEK, IT'S
SIGNIFICANTLY HIGHER IN CULTURE,
YEAH, WE'VE WORKED WITH OXYGEN
SCAVENGERS SO LOW OXYGEN AND SO
LOW EXGEN IS MUTATIONS BUT ALSO
GROWS SLOWER SO PEOPLE THAT
BELIEVE THAT STEM CELLS LIKE
HIPOXIC CONDITIONS AND IT IS
TRUE THAT SUSTAINED FOR THE
MUTATIONS BUT ALSO IF THEY
UNDERGO, IF YOU CALCULATE, YOU
DON'T WIN, BECAUSE THEY GROWTH
FACTOR SLOWER AND THEY HAVE TO
GROW LONGER AND THEY HAVE THOSE
MUTATIONS BUT IT'S CLEAR THAT
OUR--THE CULTURAL ENVIRONMENT IS
MORE MORPH O GENIC THAN WHAT
THEY SUSTAIN AND REAL LIFE,
YEAH.
>> GOOD AND THEN THERE ARE
MULTIPLE MESSAGES ABOUT WHAT A
WONDERFUL AND INFORMATIVE TALK
IT WAS AND SIGNIFICANT AND THANK
YOU, THANK YOU, THANK YOU, SO I
YOU WILL PROBABLY BE ABLE TO SEE
THOSE.
SO I--YOU KNOW I KNOW IT'S
10:00 P.M., THERE, I FEEL LIKE
WE SHOULD LET YOU GO, BUT I
WOULD JUST LIKE TO THANK YOU
AGAIN.
IT WAS A WONDERFUL PRESENTATION.
>> OKAY.
>> WE ALL LEARNED A LOT.
>> WELL, THANKS TO ALL FOR
LISTENING.
AND GOOD REST OF THE DAY DAY.