MY NAME IS MICHAEL GRAHAM ESPEY
PROGRAM DIRECTOR NCI
DIVISION OF CANCER BIOLOGY.
IT'S A PLEASURE TO INTRODUCE
WALS LECTURE SPEAKER
NAVDEEP CHANDEL.
NAV HIS FRIENDS CALL HIM
HE RECEIVED HIS Ph.D. AND POST
DOCTORAL TRAINING AT UNIVERSITY
OF CHICAGO IN CELLULAR
PHYSIOLOGY, WITH PAUL SHOEMAKER
AND CRAIG THOMPSON.
HE'S CURRENTLY THE DAVID
DISTINGUISHED PROFESSOR OF
MEDICINE AT NORTHWESTERN
UNIVERSITY AND IS NCI
OUTSTANDING INVESTIGATOR
AWARDEE.
NAV BEGAN STUDIES ON MOLECULAR
CELLULAR BASIS OF OXYGEN SENSING
WHICH NATURALLY LED HIS
CURIOSITY TO AN EVER DEEPENING
UNDERSTANDING OF MITOCHONDRIAL
METABOLISM IN DIVERSE CELL
TYPES.
BEGINNING MOST 20 YEARS AGO TO
THE PRESENT DAY, NAV DEVELOPED
THE CONCEPT OF MITOCHONDRIA
SERVING BEYOND THE MORE
TRADITIONAL VIEWPOINT OF
BIOSYNTHETIC AND BIOENERGETIC
ORGANELLES TO INCLUDE A THIRD
DISTINCT ROLE AS SIGNALING HUBS
WHEREBY PHYSIOLOGIC RELEASE OF
REACTIVE OXYGEN SPECIES AND
METABOLITES REGULATE
TRANSCRIPTION FACTORS
EPIGENETICS AND TO CONTROL
DIVERSE CELLULAR FUNCTIONS.
HIS RECENT WORK FOCUSED ON THE
SENSUALITY OF MITOCHONDRIA
METABOLIC AND RA S AGONALLING TO
CONTROL STEM CELL -- INVOLVING
CONTROL OF CANCER CELL PHENOTYPE
AND ADAPTIVE IMMUNITY AS YOU
WILL SEE IN THE PRESENTATION
TODAY.
IN ADDITION TO BEING A CHAMPION
OF BASIC SCIENCE, NAV RECENTLY
LEVERAGED HIS TRANSDISCIPLINARY
UNDERSTANDING OF THE
MITOCHONDRIA TOWARDS A MORE
TRANSLATIONAL REPURPOSING OF THE
ANTI-DIABETES DRUG METAPHORMAN
AS POTENTIAL CANCER THERAPEUTIC
SO WITHOUT FURTHER ADIEU I WOULD
LIKE TO INVITE OUR FRIEND NAV
CHANDEL UP TO GIVE TODAY'S WALS
LECTURE.
[APPLAUSE]
THANKS AGAIN, MICHAEL AND THE
FOLKS AT THE NCI AS WELL AS THE
NIH HERE FOR THIS REAL HONOR TO
GIVE A WALS LECTURE.
AS YOU CAN SEE FROM MY TITLE,
IT'S -- GOING TO TALK
MITOCHONDRIA AND HOW WE THINK
ARE ESSENTIAL PLAYERS IN NORMAL
PHYSIOLOGY AND CONTROL OF
HOMEOSTASIS AS WELL AS HOW
MITOCHONDRIA CAN BE CO-OPTED IN
DIFFERENT PATHOLOGIES.
MUCH WHAT I'M GOING TO TALK TO
YOU ABOUT IS THE TITLE SAYS
BEYOND ATP.
HOPEFULLY YOU'LL SEE WHY I THINK
THAT.
THE MOST IMPORTANT SLIDE IS THIS
ONE BECAUSE THE WORK THAT I'M
GOING TO PRESENT IS AN
UNPUBLISHED STORY FROM SAM
WINEBERG, AN M.D. Ph.D.
STUDENT IN MY LABORATORY AND
HE'S HELPED BY TWO OTHERS IN THE
LABORATORY ELIZABETH STANDARD
AND -- WE TALKED PREVIOUS DATA
FROM GREAT COLLABORATOR IT IS
OVER THE YEARS AND CURRENT AS
WELL BEN SINGER PAUL SHOEMAKER
AND CARLOS MARTINEZ WHO IS
HELPING US DO SOME OF OUR RNA
SEQUENCING AND LARRY TURKA WHO
IS WELL KNOWN FOR HIS WORK IN
T-CELLS HAS BEEN GIVING US
FRIENDLY ADVICE AND TUTORING
ABOUT T-CELL BIOLOGY.
SO WE HAVE A VERY SIMPLE
QUESTION IN THE HAS BEENTORY.
WHICH IS WHY ANY -- LABORATORY
WHICH IS WHY ANY MAMMALIAN
ASPIRES. S. IT'S ALMOST AN OBVIOUS
QUESTION, YOU HAVE A
MITOCHONDRIA, IT USES OXYGEN,
GENERATES ARCTP AND YOU ASSUME
THAT'S THE ONLY REASON WHY WE
HAVE MITOCHONDRIA.
HERE ARE CELLS WHICH ARE FOUND
IN THE GENERAL TUMOR
MICROENVIRONMENT THAT WE'RE
INTERESTED IN.
WE STARTED MUCH OF OUR WORK IN
TUMOR CELLS, BUT IT OCCURRED TO
US THAT THE TUMORS HAVE T-CELLS
AROUND MACROPHAGES ENDOTHELIAL
CELLS, AND THEN ALSO WE STARTED
TO THINK WHAT ABOUT OTHER
PROLIFERATING CELLS, THE STEM
CELLS.
WHAT IS THE METABOLISM OF NORMAL
CELLS.
THIS IS SORT OF A QUESTION THAT
WE HAVE BEEN INTERESTED IN.
JUST TO GET YOU ALL CAUGHT UP ON
THE SIMPLE BIOCHEMISTRY OF
MITOCHONDRIA, AS MANY OF YOU
KNOW THERE'S RESPIRATORY CHAIN,
THE RESPIRATORY CHAIN ELECTRONS
TO MOLECULAR OXYGEN, THIS IS
COUPLED TO A PROTON MOTOR FORCE
WHICH GENERATES ARCTP.
EVERYBODY IS COMFORTABLE WITH
THIS IDEA, IT'S TRUE, IT STILL
HOLDS TRUE, LOTS OF CELLS DO
THIS.
THE SECOND THING WHICH WAS
ACTUALLY THE ORIGINAL FUNCTION
OF MITOCHONDRIA, WELL BEFORE
OXIDATIVE PHOSPHORYLATION, WAS
WORKED OUT, IS THAT THE TCA
CYCLE METABOLITES, AND THE TCA
CYCLE IS COUPLED TO THE
RESPIRATORY CHAIN BECAUSE THE
TCA CYCLE USES NAD AND FAD AND
BECOME FADH AND NADH AND THE
ELECTRON TRANSPORT RESPIRATORY
CHAIN WILL MAKE IT BACK THE NAD
TO KEEP THE CYCLE GOING. THESE
METABOLITES ARE THE TCA CYCLE
ARE IMPORTANTTOR MACRO MOLECULE
SYNTHESIS.
IF YOU GO BACK TO GREAT
BIOCHEMISTRY BOOKS LIKE LENINGER
AND OTHER BOOKS FOLLOW THE
CARBONS, HOW DO YOU GET TO A
NUCLEOTIDE, HOW YOU GET THE A
LIPID, HOW GET TO PROTEINS,
AMINO ACIDS MUCH BACKBONES COME
IN PART FROM THE CYCLE.
HISTORICALLY THESE ARE THE TWO
FUNCTIONS THAT HAVE BEEN
ASCRIBED TO MITOCHONDRIA, THERE
TO PROVIDE YOU ENERGY, AND TO
GIVE YOU METABOLITES FOR GROWTH.
HOWEVER, WE ARGUED THERE'S A
THIRD FUNCTION, IT SEEMS OBVIOUS
TODAY BUT WHEN WE FIRST THOUGHT
ABOUT THIS MORE THAN 20 YEARS
AGO WE HAD A LITTLE BIT OF A
PUSH BACK, AND THE REASON WAS
THE WAY MANY OF US WERE TAUGHT
BIOLOGY AND WHERE MITOCHONDRIA
SITS WITHIN A CELL IS THE
FOLLOWING.
IF YOU HAVE A PHYSIOLOGIC
SIGNAL, IF FOR EXAMPLE A T-CELL
RESPONDS TO ANTIGEN OR GROWTH
FACTORS THERE'S TO PROLIFERATE
OR CELLULAR DIFFERENTIATION, ANY
PHYSIOLOGICAL PROCESSES AT THE
CELLULAR MOLECULAR LEVEL, WE'RE
HARD WIRED TO THINK THOSE
SIGNALS ARE INTEGRATED
BUSYINGNAL TRANSDUCTION
MECHANISMS, FUNNELING TO G
TRANSCRIPTIONAL NETWORKS AS WELL
AS REMODELING THE CHROMATIN,
THAT CELL WILL DO WHATEVER IT'S
SUPPOSED TO DO WHETHER
PROLIFERATE, DIFFERENTIATE, MAKE
CYTOKINES, ET CETERA.
THAT MODEL, ONCE ALL OF THIS
ACTION IS HAPPENING AT THE
SIGNALING LEVEL AND THE GENE
TRANSCRIPTION CHROMATIN
REMODELING THEN IT JUST FEEDS
BACK TO MITOCHONDRIA AND SAYS
I'M GOING TO DOUBLE MYSELF
PROLIFERATE OR DIFFERENTIATE OR
MAKE THESE CYTOKINES, GIVE ME
WHATEVER ENERGY OR BIOSYNTHESIS
CAPACITY I NEED.
THIS IS ALMOST IN A WAY A
PASSIVE PLAYER IN THIS PROCESS
JUST RESPONDING TO DOWNSTREAM
SIGNALING, GENE EXPRESSION OF
CHROMATIN REMODELING.
FOR US IT DIDN'T MAKE SENSE
BECAUSE THIS WOULD BE LIKE
GETTING IN YOUR CAR EVERY
MORNING AND NOT KNOWING HOW MUCH
FUEL YOU HAVE.
SO ONE SIMPLE THING WE THOUGHT
OF, PERHAPS MITOCHONDRIA NOT SO
PASSIVE BUT ACTUALLY PART OF THE
ACTIVE DECISION MAKING PROCESS,
THAT TO DETERMINE CELL FATE AND
CELL FUNCTION.
OPPORTUNISTIC WAY OF THINKING
ABOUT THIS, IF YOU HAVE THE
SIGNALS, YES THEY ACTIVATE MANY
SIGNAL TRANSDUCTION PATHWAYS AND
TRY TO REMODEL CHROMATIN IN
GENES BUT THEY HAVE TO GO
THROUGH OBLIGATORY STEP WHICH IS
TO ASSESS MITOCHONDRIAL FITNESS.
SIGNALS HAVE TO COMMUNICATE FROM
THE MITOCHONDRIA ON TO THE
NUCLEUS.
AND WHAT WOULD THESE SIGNALS BE?
WE GOT INSPIRED BY TWO FINDINGS,
WHEN WE REPORTED THIS PAPER ON
HYPOXIA, CONTROL OF THE HIPS,
WHICH IS A DOMINANT
TRANSCRIPTION FACTOR HYPOXIA
WHAT THE SIGNAL COULD BE SO WHEN
THE COLLEAGUES FOUND CYTOCHROME
C RELEASE, THAT HAPPENED UNDER
CELL DEATH WE START TO THINK
MITOCHONDRIA MUST BE RELEASING
THINGS.
THAT SIGNAL WAS A DEATH SIGNAL
AND WE THOUGHT MITOCHONDRIA
OBVIOUSLY ARE SO SELECTIVE FOR A
POSITIVE SIGNAL BENEFICIAL
SIGNAL, ONE OBVIOUS WAS REACTIVE
OXYGEN SPECIES AND LUCKILY HERE
FROM KOREA, ONE OF THE PIONEERS
WHO PROPOSE THAT EARLY WORK
SHOWING REACTIVE OXYGEN SPECIES
COULD BE A SIGNALING MOLECULE
AND I THINK THAT BIOLOGY
CONTINUES TO EXPAND AND OTHER
WORKING ON THIS PROPOSE NAD,PH
OXIDASE BEING A CRITICAL
GENERATOR OF ROS.
WE THOUGHT MAYBE THE
MITOCHONDRIA COULD BE THAT
GENERATOR OF ROS.
PEOPLE DIDN'T LIKE THAT IDEA
BECAUSE THEIR SIMPLISTIC NOTION
MITOCHONDRIA ONLY GENERATED ROS
WHEN DAMAGED.
THIS EXPLAIN NEURODEGENERATION,
MAYBE THIS EXPLAIN HEART FAILURE
FREE RADICAL THEORY OF AGING AND
THIS IDEA THAT MITOCHONDRIA ARE
BAD ACTORS.
AND SO ONE OF THE THINGS WE
CONTINUE TO SHOW OVER THE YEARS
WAS THAT'S NOT QUITE TRUE.
MANY INSTANCES WE THINK
MITOCHONDRIA RELEASE REACTIVE
OXYGEN SPECIES UNDER
PHYSIOLOGICAL STIMULI TO CONTROL
A VARIETY OF IMPORTANT
TRANSCRIPTIONAL NODES INCLUDING
NOTCH SIGNALING NF KAPPA B.
SO THE RESPIRATORY CHAIN ISN'T
PERFECT AND ELECTRONS CAN LEAK
TO MAKE SUPER OXIDE AND HYDROGEN
PEROXIDE TO MAKE MITOCHONDRIAL
REACTIVE OXYGEN SPECIES RELEASED
UNDER VARIETY OF STIMULUSES, TO
CONTROL THESE IMPORTANT
TRANSCRIPTIONAL NETWORKS.
THE OTHER THING, MORE THAN A 15,
20 YEARS AGO WE -- EPIGENETIC
AND CHROMATIN FIELDS WERE
DISCOVERING ENZYME OVER THE PAST
TWO DECADE LIKE HISTONE
ACETYLASES THAT CONTROL
CHROMATIN REMODELING.
ONE THING THAT WAS OBVIOUS TO
SOMEONE WHO WORKS ON
MITOCHONDRIA, ISN'T IT
INTERESTING THEY USE TCA CYCLE
METABOLITES TO WORK.
WHERE DOES YOUR ACETYL COA COME
FROM TO MAKE DEMETHYLASES WORK
AN THINGS LIKE SUCCINATE ARE
POWERFUL INHIBITORS OF THESE
DEMETHYLASES.
I HAVE QUITE A WONDERFUL
COLLEAGUE AT NORTHWESTERN
INCLUDING BIOCHEMISTRY CHAIR
ALI, MANY OF YOU KNOW HIS WORK
IN THE CHROMATIN FIELD, HE'S
LOOKING AT OUTPUT OF
MITOCHONDRIA WHEN HE STUDIES
EPIGENETICS BECAUSE THESE THINGS
WE HAVE BEEN ABLE TO TCA CYCLE
METABOLITES, CONTROLLING THE
HISTONE ACETYLATIONS.
IF WE RESTRICT GENERATION OF
CITRATE HERE WE CAN IMPACT
HISTONE ACETYLATIONS AND WE
PUBLISHED THIS BEFORE.
IN THINKING ABOUT WHAT
FUNCTIONAL LIVE MITOCHONDRIA DO,
WE CAN MANY OF US ARE
COMFORTABLE MITOCHONDRIA MIGHT
BE IMPORTANT FOR SURVIVAL BY
KEEPING THE ATP ADP RATIO HIGH
TO DRIVE MANY CELLULAR
REACTIONS.
GLYCOLYSIS CAN COMPENSATE
BECAUSE THEY GENERATE ATP.
THE OTHER, IT'S IMPORTANT FOR
LIPIDS NUCLEOTIDES, MANY MACRO
MOLECULE SYNTHESIS COMES FROM
MITOCHONDRIA SO IT'S
THIRD FUNCTION MY LAB IS REALLY
WORKED A LOT ON IS THIS IDEA
THERE'S THINGS RELEASED FROM
MITOCHONDRIA, METABOLITES,
REACTIVE OX GENERAL SPECIES
WHICH CONTROL TRANSCRIPTION
THROUGH CHROMATIN REMODELING
DIRECTLY BY ACTIVATING
TRANSCRIPTION FACTORS.
PLAYS OUT DIFFERENTLY IN
DIFFERENT CELL TYPES IN VIVO.
SEEMS LIKE CANCER CELLS USE
MITOCHONDRIAL METABOLISM NOR
GROWTH AND SURVIVAL.
WHERE STEM CELLS IN PARTICULAR
HEMATOPOIETIC STEM CELLS DON'T
CARE ABOUT ABILITY TO
PROLIFERATE.
IF WE KNOCK OUT THE SAME
RESPIRATORY CHAIN GENES AND
CANCER CELLS AND STEM CELLS,
THEY CONTINUE TO PROLIFERATE,
THE FETAL STEM CELLS,
HEMATOPOIETIC PROLIFERATE BUT
NOT DIFFERENTIATE INTO
PROGENITORS.
ADULTS WHICH TEND TO BE
QUIESCENT LOSE ITS AND UNDERGO
HYPERPROLIFERATION AND STEM CELL
EXHAUSTION SHUN.
COMPLETELY DIFFERENT IN THE
CANCER.
WHY THAT IS, WE'RE STILL TRYING
TO FIGURE OUT AND FINALLY
T-CELLS, SEEMS TO BE ALL ABOUT
THEIR ABILITY TO FUNCTION SO IF
IT'S A REGULATED T CELLS, IT
DOESN'T SUPPRESS.
I'LL SHOW YOU WHAT I MEAN.
I CAN'T GO INTO ALL THE MODEL
SYSTEMS BUT SIMPLE COMPARE AND
CONTRAST IN VIVO I'LL FOCUS ON
CANCER AND ON REGULATORY T-CELLS
TO SHOW AN EXAMPLE.
WE GOT INTO THE CANCER
METABOLISM FIELD, MORE THAN 12,
13 YEARS AGO, AS YOU KNOW CANCER
METABOLISM FIELD IS REVIVED IN
PART BY OLD OBSERVATION MADE BY
OTTO WARBERG ALMOST 90 YEARS AGO
WHICH SHE OBSERVED A CANCER
TISSUE UNDER NORMAL OXYGEN
CONDITIONS RIGHT ON THE
LABORATORY BENCH WOULD MAKE LOTS
OF LACTATE.
THIS IS CALLED THE AEROBIC
GLYCOLYSIS OR THE WARBERG
EFFECT.
CLINICIANS KNOW THAT USING PET
IMAGERY YOU CAN SEE WITH A
GLUCOSE TRACER PET POSITIVE.
SO YES, TUMORS O GLYCOLYTIC.
THERE'S A LOT OF HYPE AROUND
THAT TO THE POINT PEOPLE THINK
THE MITOCHONDRIA DISPENSABLE IN
CANCER.
SO WE SAID DOESN'T MAKE SENSE
BASED ON SIMPLE BIOCHEMISTRY
KNOWLEDGE AND WE INVESTED EARLY
ON IN MAKING MOUSE MODELS USING
MOUSE MODELS OF CANCER LIKE
TYLER JACKS AND OTHERS PIONEERED
BUT ALSO MAKING SOME MODELS WE
CAN USE IN VIVO TO TEST
NECESSITY OF THE RESPIRATORY
CHAIN SO ONE WE MADE WAS TFAM.
SO TFAF ENCODES, IS A NUCLEAR
PROTEIN NECESSARY FOR
MITOCHONDRIA DNA REPLICATION AND
TRANSCRIPTION SO REMEMBER THE
MITOCHONDRIA HAS ITS OWN GENOME
AND IT ENCODES FOR CERTAIN KEY
SUBUNITS OF RESPIRATORY CHAIN.
IF YOU -- WE GENERATE A PHLOX
MOUSE, WHEREVER YOU KNOCKED OUT
MITOCHONDRIAL DNA IS GONE.
MITOCHONDRIAL DNA IS GONE, A
VARIETY OF THE CRITICAL SUBUNITS
OF THE RESPIRATORY CHAIN ARE
GONE.
YOU DON'T HAVE MITOCHONDRIA THAT
ASPIRES.
FUNCTIONALLY IN CELL BIOLOGY
EVEL IF YOU LOOK BY ELECTRON
MICROSCOPY THE MITE CON TRIA
THERE THAT DON'T RESPIRE.
SO WE ASKED WHAT HAPPENS IN ONE
OF THESE MOUSE MODELS THAT THE
FIELD USES AND THE ONE WE USE IS
THE LOCK STOP LOCKS KRAS
ONCOGENIC KRAS DRIVEN LUNG
CANCER MODEL WE DUMP ADENOVIRUS,
CRE AND THE LOCK STOP LOX ALLELE
IS GONE ACTIVATES ONCOGENIC KRAS
AND YOU SEE INCREASE IN IN TUMOR
BURDEN.
TO THE E TENT YOU KNOCK OUT TFAM
AND EXCISE THE TFAN GENE WE
NOTICE LESS TUMORS.
SO REMEMBER BY KNOCKING OUT TFAN
WE HAVE MADE THE TUMORS
COMPLETELY DEPENDENT ON
GLYCOLYSIS.
THE ULTIMATE WARBERG CELLS AS I
CALL THEM BUT LESS TUMORS THIS
SIMPLE EXPERIMENT TOLD US WITH
NECESSITY OF THE RESPIRATORY
CHAIN UNDER THESE MOUSE MODELS,
FOR TUMORIGENESIS.
ONE THING WEES ARE GOT INTO WAS
METAPHOR MAN BECAUSE
EPIDEMIOLOGISTS NOTICED THAT
PEOPLE WHO HAD TAKEN METFORMAN
FOR DIABETES HAS LOWER CANCER
ACROSS SOME CANCERS THIS LED TO
CLINICAL TRIALS INCLUDING LARGE
SCALE HAPPENING IN CANADA IN
BREAST CANCER.
AND THE RESULTS WILL BE OUT NEXT
YEAR.
BUT SOMETIMES WHEN A DRUG IS
REALLY CHEAP AND EASILY
REPURPOSED AND OBVIOUSLY
UNFORTUNATELY PATIENTS SUFFERING
FROM THE TERRIBLE DISEASE OF
CANCER YOU RUSH TO DO THE
CLINICAL TRIALS.
AND I'M NOT COMPLETELY CONVINCED
WE HAVE BEEN VERY I SHOULD SAY
VERY THOUGHTFUL ABOUT SOME OF
THESE TRIALS, BECAUSE IT'S NOT
CLEAR WHETHER THE ANTI-DIABETIC
DOSE SHOULD BE THE DOSE FOR
ANTI-CANCER.
IN FACT, OUR STUDIES MIGHT
SUGGEST YOU NEED ALMOST A
MAXIMAL TOLERATED DOSE OF
METFORMAN TO HAVE EFFICACY.
NOT EVERY TUMOR RESPONDS BECAUSE
IN ORDER FOR IT TO GET INTHE A
TUMOR CELL IT NEED ORGANIC
CATION TRANSPORTERS.
IN THE ABSENCE OF THAT
TRANSPORTER YOU CAN THROW
BUCKETS MILLIMOLAR AMOUNTS AND
NOTHING HAPPENS.
OTHER QUESTION IS WHAT'S THE
TARGET?
IN 2000 THERE WAS TWO REPORTS IN
JBC THAT SUGGESTED GIVE ISOLATED
MITOCHONDRIA METFORMAN YOU CAN
INHIBIT COMPLEX 1.
THERE'S 45 SUBUNITS IN COMPLEX
1.
THE BEST WAY TO SHOW WHAT A DRUG
DOES, THE GOLD STANDARD IS FIND
IT WHERE IT BINDS IN PARTICULAR
COMPLEX OR PROTEIN SO IT DOESN'T
CHANGE THE CATALYSIS OF THAT
PROTEIN BUT MAKES IT REFRACTORY
TO BINDING, THAT'S THE GOLD
STANDARD.
I'M NOT A STRUCTURAL BIOLOGIST
WE DIDN'T DO ELEGANT EXPERIMENT
LIKE THAT.
IT'S A 45 SUBUNIT COMPLEX.
PRETTY BIG BUT WE DID SOMETHING
CLEVER, WE SAID LET METAPHORMAN
INHIBIT MITOCHONDRIAL COMPLEX 1
AND IF NECESSARY FOR ANTITUMOR
EFFECTS LET'S RESCUE IT BY
PUTTING IN THE CANCER CELLS A
PROTEIN CALLED NDI 1, FOUNDING
EAST.
SACK CROW MYSISTER SRIS YEAH.
NAD, THE ONLY THING IT DOESN'T
DO IS PUMP HYDROGEN.
AND BY PUMPING -- DOING NADH TO
NAD DONATE ELECTRONSES TO COQ
AND DOWN TO MOLECULAR OXYGEN.
SO FUNCTIONAL RESCUE.
WE DID THIS IN A COLON CANCER
MODEL AND LUNG CANCER MODEL.
AS YOU CAN SEE WE LET THE TUMORS
GROW, IF WE GIVE METAPHORMAN IN
THE DRINKING WATER WE CAN
INHIBIT TUMORIGENESIS, IF THERE
ARE NDI 1 THERE THEY'RE
REFRACTORY, ALL CONTROL EXCEPT
PLUS OR MINUS NDI 1 OR
METAPHORMAN.
WHAT'S NICE IS THAT ONCE WE
PUBLISHED THIS COUPLE OF YEARS
AGO, A BUNCH OF OTHER PHYSICIAN
SCIENTISTS AND CLINICIAN
SCIENTISTS HAVE NOW GONE AND
GIVEN METFORMAN TO PATIENT
COHORTS AND TAKEN BIOPSIES TO DO
METABALOMICS AND WE KNOW A
SIGNATURE WHEN MITOCHONDRIAL
RESPIRATORY CHAIN INHIBITED WE
KNOW A METABOLIC SIGNATURE
CONSISTENT WITH THAT AND NOW
GIVING METAPHORMAN TO PATIENTS
YOU CAN SEE THAT SIGNATURE
IMPLYING METFORMAN IN HUMAN
PATIENTS CAN GET INTO THEIR
CANCER CELL MITOCHONDRIA AND
PERTURB METABOLISM.
SO ONE OF THE THINGS WE HAVE
BEEN TRYING TO FIGURE OUT WHAT
ASPECTS OF THE RESPIRATORY CHAIN
ARE CRITICAL.
DO YOU NEED ATP COMING, NOR IS
IT JUST ABOUT THE METABOLITES.
ONE WAY WE THOUGHT ABOUT DOING
THIS IS WE KNOCKED OUT
MITOCHONDRIA COMPLEX 3, KNOCK
OUT COMPLEX 3 IN THE MIDDLE
THESE ELECTRONS AREN'T GOING TO
FLOW AND YOU WON'T PROTON PUMP,
TCA CYCLE WON'T WORK OR
CANONICALLY LIKE IT SHOULD AND
THEN MAYBE YOU WANT TO GROW.
SO IF YOU KNOCK OUT COMPLEX 3
CONSISTENT WITH WHAT EVERYTHING
ELSE I HAVE SHOWN YOU, AGAIN,
YOU GET VERY LITTLE TUMORS
GROWING OUT.
NOW AGAIN, WE HAVE GOT INTO
GIVING ANCIENT ENZYMES TO
COMPLIMENT MUTATIONS.
HERE IS ANOTHER ONE WE AGAIN
TAKE THE COMPLEX 3 KNOCK OUTS,
ALL GONE, HERE WE PUT A PROTEIN
CALLED THE ALTERNATIVE OXIDASE
FOUND IN PLANTS, FOUND IN SEA
AND LOWER ORGANIZE IMS, IT CAN
TAKE ELECTRONS FROM Q AND AGAIN,
TRANSFORM TO MOLECULAR OXYGEN.
SIMILAR TO CYTOCHROME C OXIDASE
BUT WILL ALLOW THE TCA CYCLE TO
OCCUR BECAUSE COMPLEX 1 AND 2
ACTIVITY IS REGAINED BECAUSE IT
CAN DONATE ALL THIS ELECTRONS SO
WHEN YOU DO THAT, COMPARED TO
THE CONTROL, COMPLEX 3 KNOCK
OUTS THAT DON'T GROW PEER BY
PUTTING BACK AOX WE CAN RECOVER
TUMORIGENESIS.
THIS SIMPLE EXPERIMENT STARTS TO
POINT ONE OF THE KEY FUNCTIONS
OF MITOCHONDRIA WE THINK IS TO
GENERATE THOSE TCA CYCLE
METABOLITES.
IT'S FOR GROWTH AND A MODEL
THAT'S EMERGED FROM MY LAB AND
PEOPLE AT RALPH DENNIS WHO HAD
DONE NICE WORK IN HUMAN PATIENTS
WITH GLUCOSE TRACERS, AS WELL AS
WORK OF ALEX KIMMELMAN AND
OTHERS, HAS COME TO -- THIS IS
IS A BIRD'S EYE VIEW THAT IN THE
LAND OF PLENTY WHEN NUTRIENTS
ARE AVAILABLE THE MITOCHONDRIA
IS BIOSYNTHETIC BIOENERGETIC HUB
ALIGNED FOR RAPID CELL
PROLIFERATION.
IT CAN GIVE YOU ALL THE LIPIDS,
HELP MAKE NUCLEOTIDES, GIVE YOU
THE ENERGY, BUT MANY OF YOU
KNOW, TUMORS SIT IN HARSH
ENVIRONMENTS AND THERE MANY WAYS
CANCER METABOLISM COMMUNITIES
DISCOVERED, WE HAVEN'T
CONTRIBUTED MUCH TO THIS, WHICH
IS THINGS LIKE CYTOSIS OR
AUTOPHAGY OR BRANCHING AMINO
ASITS OR FATTY ACID OXIDATION,
ALL THESE SURVIVAL PATHWAYS ONE
OF THEIR JOBS IS TO CATABOLIZE
WHATEVER THEY CAN IN A CELL TO
FEED THIS MITOCHONDRIA MAKE
ENOUGH ENERGY TO AT LEAST
SURVIVE UNTIL YOU GET NEW
VASCULARIZATION AND GO BACK TO
THE LAND OF PLENTY, TO GROW.
SO UNDER BOTH CONDITIONS,
NUTRIENT REPLETE CONDITION IS
ANN BOLLIC MACHINERY AND
NUTRIENT DEPLETED IS A CAT
BOLLIC MACHINERY TO GENERATE ATP
FOR SURVIVAL.
THIS IS A MODEL WE CONTINUE TO
TEST AND OBVIOUSLY WE'RE LOOKING
FOR NODES THAT WE CAN INTERFERE
WITH FOR CANCER THERAPY.
SO REALLY THE TAKE HOME HERE IS
THAT WE THINK IN VIVO ROLE OF
THE RESPIRATORY CHAIN, IS
CONTROL PROLIFERATION SURVIVAL,
I WON'T HAVE TIME TO SHOW YOU
THE STEM CELL DATA BUT WE CAN DO
SIMILAR GENETIC TRICKS KNOCKING
OUT COMPLEX 3 OR OTHER WAYS TO
KNOCK OUT THE RESPIRATORY CHAIN
AS I ALLUDED TO, STEM CELLS
THEMSELVES ARE THERE, THEY'RE
PHENOTYPICALLY LOOK LIKE STEM
CELLS BUT DON'T DO WHEY THERE
SUPPOSED TO WHICH IS GENERATE
PROGENITORS.
SO THEY DON'T HAVE A
PROLIFERATIVE DEFECT LIKE CANCER
CELLS IN VIVO BUT THEY HAVE A
DIFFERENTIATION DEFECT.
AND THE T-CELLS I'M GOING TO
SHOW YOU, IT'S ABOUT THEIR
FUNCTION.
SO WE GOT INTO THIS SEVEN OR
EIGHT YEARS AGO, TALENTED M.D.
Ph.D., WE WERE WATCHING THE
FIELD OF IMMUNOMETABOLISM THAT
CONTINUES TO GROW AND EVOLVE AND
AGAIN, THERE HISTORIC BIAS HAS
ALWAYS BEEN THEY'RE GLYCOLYTIC.
IN FACT RAPIDLY PROPROLIFERATING
CELLS SHOW PROBUST GLYCOLYSIS
BUT IT SHOULDN'T BE
MISINTERPRETED THEIR
MITOCHONDRIA ARE NOT EQUALLY
IMPORTANT.
SO AGAIN, WE DO VERY SIMPLE
EXPERIMENTS, WE ASK THE QUESTION
IS THE RESPIRATORY CHAIN
NECESSARY FOR PROLIFERATION OR
T-CELL ACTIVATION?
AND COULD WE USE SOME OF OUR
NEWER MOUSE MODELS?
ONE OF THE ONES WE HAVE BEEN
USING IS KNOCKING OUT CATALYTIC
SUBUNIT OF COMPLEX 3, IT HAS 11
SUBUNITS, THE RISKY IRON SULFUR
PROTEIN IS ENCODED BY NUCLEI
EIUS SO WE CAN PHLOX THESE
ALLELES KNOCK OUT COMPLEX 3
CROSS IT TO CD4 CRE AND THE
RESPIRATORY CHAIN WON'T WORK.
IF IT DOESN'T WORK YOU DON'T
MAKE ATP OBVIOUSLY.
SO B CELLS BECOME PURELY
GLYCOLYTIC.
GROWTH MAYBE COMPROMISED BECAUSE
YOU MAY NOT MAKE ENOUGH CYCLE
METABOLITES.
THE ROSS GENERATED BY COMPLEX 3
FOR SIGNALING MIGHT BE AFFECTED.
METABOLITES FOR HISTONE
ACETYLATIONS MIGHT BE AFFECTED
SO MULTIPLE EFFECTS AND IT GETS
DIFFICULT TO DISCERN ALL THE
REASONS WHY.
BUT LAURA INITIALLY DID THIS ON
THE SIMPLE EXPERIMENTS ONE OF
THEM IS YOU CAN TAKE A RAG
DEFICIENT MOUSE, LYMPHOPENIC
MOUSE AND ADOPTIVELY TRANSFER
WILD TYPE OR RISK KNOCK OUT THIS
IS THIS HOMEOSTATIC EXPANSION OF
T-CELLS, VERY BREAD AND BUTTER
ASSAY OF COMMUNITY.
SHE NOTICED NO PHENOTYPE, THE
WILD TYPE CELLS, IF YOU WILD
TYPE T-CELLS PUT THEM IN THIS
RAG DEFICIENT MOUSE, IT
PROLIFERATES FINE, AS OTHERS
HAVE SHOWN.
AND SO DO OUR COMPLEX 3 KNOCK
OUTS.
WE SUGGESTED THAT THERE'S NO
GROWTH DEFECT.
I DID REMIND LAURA THAT T-CELLS
ARE THERE TO RESPOND, SHE SHOULD
TRY THE VARIETY OF ANTIGENS WHEN
SHE DID THAT THEY GOT THE
OPPOSITE RESPONSE SO THEY HAVE
THE ABILITY TO GROW UNDER
HOMEOSTATIC CONDITIONS AND JUST
DON'T DO IT WHEN THEY HAVE TO
RESPOND TO AN ANTIGEN.
THAT THAT STARTED TO SMELL LIKE
SIGNALING BECAUSE THEY CAN FIND
A WAY TO GROW AND SURVIVE UNDER
ONE CONDITION BUT NOT THE OTHER.
WITH THAT GOING THROUGH A LOT OF
MECHANISTIC DETAILS AS THIS WAS
PUBLISHED A WHILE BACK, I WANT
TO SHOW YOU MORE RECENT
UNPUBLISHED DATA, BUT THIS IS
SORT OF WAS A NICE EXPERIMENT.
SO REMEMBER THE COMPLEX 3 KNOCK
OUTS ARE GLYCOLYTIC SO YOU CAN
ARGUE THE FACT THEY'RE TOO
GLYCOLYTIC OR SOMETHING ABOUT
ENERGY, THAT WAS A PROBLEM.
BUT WE LIKE SIGNALING PARADIGM,
ONE OF THE WAYS WE THINK
SIGNALING IS GENERATING
MITOCHONDRIAL ROS SO LOOK AT IL
2 PRODUCTION THIS IS IN CELL
CULTURE THE WILD TYPE CELLS WHEN
STIMULATED MAKE IL 2, THE
CLASSICAL SITESKINE OF T-CELLS
BEINGS ACTIVATED.
THE COMPLEX 3 KNOCK OUT RISK
KNOCK OUT MARKED REDUCTION.
AND ALL WE DO IS WE TITRATE IN
NANOMOLAR AMOUNTS OF GALACTOSE
OF H 2 O2 BY GIVING GALACTOSE
OXIDASE.
IT'S A SUBSTRATE FOR GALACTOSE
OXIDASE AND WE CAN TITRATE THE
SUBTRAIT AND THE -- SUBSTRATE
AND THE ENZYME CONCENTRATION AND
TO WHATEVER LEVEL WE WANT, WE
CAN GENERATE NANOMOLAR,
MICROMOLAR MILLIMOLAR TO
CONTINUOUSLY AND BY FLOODING THE
SYSTEM WITH NANOMOLAR AMOUNTS WE
ASKED WHETHER IN THE RISK KNOCK
OUT WE CAN BRING BACK THE IL 2.
AND WE CAN.
THE BLACK BARS ARE THE RISK
KNOCK OUT SO VERY LITTLE IL 2
WHEN STIMULATED AS LONG AS WE
PROVIDE ONE THING BACK.
H 2 O2 WE CAN BRING BACK IL 2
LEVELS.
THAT SUGGESTED THAT ONE OF THE
THINGS THAT WAS IMPORTANT IN
ABSENCE OF COMPLEX 3 WAS PERHAPS
ROS PRODUCTION FROM COMPLEXIN.
WE SHOWED THAT USING DYES AND
OTHER METHODS BUT THE SIMPLE
MECHANISM THAT WE OUTLINED WAS
THAT THE EARLY SIGNAL ONCE YOU
ACTIVATE THE T-CELL BY T-CELL
RECEPTOR ENGAGEMENT IS INFLUX OF
CALCIUM, ONE OF THE EARLIEST YOU
CAN DETECT AND LARGE PART THAT
SIGNAL WAS INTACT.
IN OUR KNOCK OUTS.
WHAT WASN'T INTACT WAS THE
GENERATION OF HYDROGEN PEROXIDE
IN OUR KNOCK OUTS AND THAT WAS
NECESSARY FOR TRANSLOCATION INTO
THE NUCLEUS FOR IL 2 PRODUCTION
AS MANY OF YOU KNOW N FAT IS ONE
OF THE KEY TRANS SUBSCRIPTION
FACTORS NECESSARY FOR IL 2, NF
KAPPA B AND AP 1 ARE THE OTHER
TWO.
THEY WERE NOT IN EFFECTED SO
THERE WAS SOME SPECIFICITY TO
THIS PATHWAY.
AND SO THE IDEA WE HAVE IS THAT
THE CALCIUM WOULD ENTER
MITOCHONDRIA TO ACTIVATE THE
DEHYDROGENASES, TO PUMP UP AND
REV UP RESPIRATORY CHAIN
ACTIVITY AND BY-PRODUCT IS H 2O
2 WHICH THEN ALLOW FOR THIS
INFAT DEPENDING IL 2 PRODUCTION.
WHEN SAM WINEBERG TALENTED
Ph.D. GRADUATING NEXT MONTH,
WANTED TO FOLLOW-UP AND START TO
ASK BEYOND ACTIVATION WHAT ARE
THE DIFFERENT SUBSETS?
WHAT IS ROLE OFs RESPIRATORY
CHAIN IN MEMORY CELL OR EFFECTOR
CELL OR IN A REGULATORY T-CELL
AND WE LIKE THE REGULATORY
T-CELLS BECAUSE THERE'S NICE
LITERATURE IN REGULATORY T-CELL,
ALSO MUCH OF HOW THE TREG T-CELL
LITERATURE, THERE'S EPIGENETIC
CONTROL OF TREG FUNCTION WHICH
TO US LIKE I SAID, WE LIKE TO
THINK OF CHROMATIN REMODELING
AND EPIGENETIC LANDSCAPE BECAUSE
IT'S -- WE CAN SEE HOW IT NICELY
TIES INTO MITOCHONDRIA FUNCTION
BECAUSE MANY TCA CYCLE
METABOLITES CONTROL THOSE
ENZYMES.
SO AGAIN, VERY SIMILAR APPROACH.
WE TAKE THE FOX P 3 CRE AN THESE
MICE HAVE YFP SO YOU CAN TAG
THEM IF CRE IS ACTIVE AND CROSS
TO THE COMPLEX 3.
THE RISK OF PHLOX MICE SO YOU
HAVE COMPLEX 3 GONE AGAIN AND
WHAT HAPPENS?
AND WE GOT A COMPLETELY
ASOUNDING RESULT, HE T GO A
MOUSE THAT LOOKS LIKE A FOX P 3
NULL MOUSE.
SO FOX P 3 IS THE LINEAR
SPECIFIC TRANSCRIPTION FACTOR
NECESSARY FOR REGULATORY T-CELL
FUNCTION.
YOU LOOSE FOX P 3 IN A MOUSE YOU
GET THE MOUSE AND THREE WEEKS
THE MOUSE DIES THERE'S
AUTO-IMMUNITY, IMMUNE
DISREGULATION EVERYWHERE,
HYPERINFLAMMATION AND PATIENTS
THAT HAVE MUTATIONS IN FOX P 3
ALSO HAVE THIS HYPERINFLAMMATION
IMMUNE DISREGULATION.
WE GOT THE ONLY MOUSE, I KNOW
ALMOST ALL PHENO COPIES.
I COULDN'T BELIEVE IT.
GENETICS DON'T LIE SOMETIMES
THEY DO, WE MADE SURE THAT'S
TRUE, WE SHOULD TAKE ANOTHER
SUBUNIT OF COMPLEX 3.
THAT'S 11 SUBUNITS ONE ENCODED
BY MITOCHONDRIAL DNA, THE OTHER
TEN ARE NUCLEAR ENCODED AND ALL
OF THOSE TEN ARE ALL NECESSARY
FOR THE COMPLEX.
IF YOU LOSE ANY ONE, THE COMPLEX
DOESN'T WORK WELL.
THIS IS THE QPC PROTEIN, WE GOT
IDENTICAL PHENOTYPE SO ALL THE
DATA I SHOW YOU, WE SORT OF
PHENO COPIED WITH EACH OTHER
MOUSE.
THAT WAS GOOD.
THOUGH WE WERE QUITE EXCITED BY
THIS FINDING THERE WAS ONE
SIMPLE EXPERIMENT SAM HAD TO DO
FOR ME TO GET EXCITED.
LISTEN, IF YOU SHOW THIS TO
ANYBODY THEY'LL SAY YOU HAVE A
DEAD MOUSE BECAUSE YOU HAD NO
TREGS, YOU GOT A MOUSE TREGS
NEEDED FOR SURVIVAL TELL ME WHAT
THE TREG NUMBERS ARE.
IF THEY'RE COMPLETELY GONE,
REALLY LOW, IT'S A COOL FINDING
BUT IT AIN'T THAT COOL.
YOU NEED ATP.
WHAT HE FOUND WAS THE OPPOSITE.
THE NUMBER OF FOX P 3 POSITIVE
CELLS, WERE IDENTICAL BETWEEN --
IN FACT LITTLE BIT MORE IN THE
COMPLEX KNOCK OUT.
AND YOU CAN SEE IF YOU TAKE OUT
THESE NUMBERS OUT AND PUT KI 67
OR CD4 4 WHICH IS ACTIVATION
MARKER, PHENOTYPICALLY WE HAVE
GOT PLENTY OF FOX P 3 POSITIVE
SO CALLED TREGS THAT
PROLIFERATE, THAT LOOK LIKE
THEY'RE ACTIVATED, THEY'RE
SURVIVING, PROLIFERATING.
SO WE'RE DOWN TO THE LAST
FUNCTION.
ARE THEY SUPPRESSIVE?
AND HERE I'M SHOWING YOU WE HAVE
DONE IN VITRO SUPPRESSION AWE
SAYS THEY DON'T SUPPRESS WELL.
HERE IS IN VIVO.
THIS IS WHERE AGAIN WE GO BACK
INTO RAG MICE WHERE WE GIVE WILD
TYPE EFFECTOR CELLS AND YOU CAN
SEE THESE MICE DEVELOP COLITIS
AND IMMUNE DISREGULATION AND
DIE.
IF YOU GIVE WILD TYPE EFFECTORS
WITH THE WILD TYPE TREGS, THAT
SHOULD SUPPRESS VECTOR CELLS,
CLASSIC SUPPRESSION ASSAY, THE
MICE SURVIVE.
BUT NOW REDO THE EXPERIMENT WITH
WILD TYPE EFFECTORS WITH THE
KNOCK OUT TREG, AND LOOKS LIKE
THEY NEVER GOT ANY REGULATORY
T-CELLS.
THIS TELLS US WE HAVE T-CELLS
THAT ARE THERE, PROLIFERATE AND
SURVIVE BASED ON MARKERS THAT
WOULD BE CALLED ESSENTIAL FOR
TREGS, MANY H ARE THERE
INCLUDING CTLA 4, FOX P 3 BUT
THEY DON'T SUPPRESS.
ANOTHER EXPERIMENT WE CAN DO NOW
WE GO TO INDUCIBLE SYSTEM.
INSTEAD OF USING FOX P 3 IN
DEVELOPMENT THIS IS TO MAX FEN
INDUCIBLE CRE SO WE LET THEM GET
TO ADULT AND NOW WE ADD A B 16
MELANOMA, THE CLASSIC IN MY LAB
WE DON'T TRY TO REINVENT TOO
MANY TECHNIQUES OTHER THAN OUR
MITOCHONDRIAL STUFF AND YOU GET
A NICE TUMOR THAT GROWS OUT AND
NOW WE'RE ADDING TO MOCKS FEN.
IT -- TAMOXIFEN, IT ONLY
ACTIVATES CRE IN FOX P 3 TREG
CELLS, BY KNOCKING OUT COMPLEX 3
INDUCEBLY IN THE REGULATORY
T-CELLS, OBVIOUSLY WE HAVE
ALLOWED THESE DREGS TO STOP
SUPPRESSING AND UNLEASH IMMUNITY
AND MAKE VERY SMALL TUMORS THAT
GET CLEARED OUT.
SO THAT TELLS US THAT COMPLEX 3
IS NECESSARY FOR TREG FUNCTION.
THE QUESTION IS WHY.
SO HERE, WHAT WE HAVE DONE IS
TAKEN THE TREG OUT AND SHOT
THROUGH RNA SEQ ANALYSIS TO GIVE
IDEA WHAT'S UP, WHAT'S DOWN.
THE NUMBER ONE PATHWAY WE
CONSISTENTLY SEE IN VIVO WHEN
MITOCHONDRIA ARE -- RESPIRATORY
CHAIN IS IMPAIRED IN OUR STEM
CELLS AND K CELLS SO FAR IS MIC.
IT TENDS TO BE UP AND SO DOES
MTOR 1, THOSE PATHWAYS TEND TO
BE UP.
AND IT'S ALMOST AS IF THE SYSTEM
WAS SENT SOME THINGS WRONG AND
TRYING TO COMPENSATE BY
ACTIVATING.
REMEMBER MIC AND M TOR 1 ARE TWO
OF THE PATHWAYS THAT CONTROL
METABOLISM.
REGULATES ALMOST EVERY METABOLIC
GENE PHOSPHORYL HATING
SUBSTRATES IN THE LIPO GENESIS
AND NUCLEOTIDE SYNTHESIS PATHWAY
SO THIS IS A FEEDBACK ON THE
SYSTEM.
BUT THE OTHER THING, WHAT'S DOWN
REGULATED?
WE THINK THAT IT MUST BE DOWN
REGULATING SUPPRESSIVE GENES.
IT'S TWO OBVIOUS ONES WERE FOX P
3 AND CTLA 4.
BUT THEN WE STARTED TO LOOK INTO
THIS A LITTLE BIT MORE HERE ARE
GENES THAT HAVE BEEN ASSOCIATED
TO SUPPRESS SHY REGULATORY
T-CELL FUNCTION.
IF YOU LOSE THESE GENES IN
REGULATORY T-CELLS YOU
EVENTUALLY DEVELOP AUTOIMMUNE
DISORDER.
SOME OF THESE NEUROPILL LYNN 1
TAKES MORE THAN A YEAR, SOME
LESS, BUT REMEMBER WE'RE HAVING
A WHOLE HOST OF THESE GENES ALL
COORDINATEDLY DOWN REGULATED
IT'S ALMOST LIKE WE MADE
MULTIPLE KNOCK OUTS ALL IN ONE.
SO HOW DO YOU COORDINATE
WIDESPREAD SUPPRESSIVE GENOME
DOWN REGULATION?
WHAT ARE SOME OF THESE -- HOW DO
YOU GET ALL THESE GENES PERHAPS
TO DOWN REGULATE?
WE TURN TO DNA METHYLATION AND
HISTONE METHYLATION.
THAT'S A LARGE FAMILY OF ENZYMES
CALLED ALPHA GLUTE RATE
HYDROGENASES.
MY FRIEND BILL GAVE THE WALS A
COUPLE OF WEEK AGO, PRO OWE
HYDROXYLACE.
SO THESE ARE REGULATORS OF HIF.
THEY CONTROL HYDROXYLATION OF
HIF AND THEIR ALPHA KETOGLUTE
RATE DEPENDENT.
AND SOME OF THE OTHER ONES ARE
THE GEMONGI DOMAIN, TEP ENZYMES
THAT ARE PARTICIPATING IN DE
METHYLATION.
SO THE HISTONE LYSINE
DEMETHYLATION, THE DNA
DEMETHYLATION, ALL THESE
REACTIONS ARE DRIVEN BY ALPHA
KETOGLUTE RATE.
THEY USE OXYGEN, MAKE CO 2 AND
MAKE IRON AND THE BY-PRODUCT IS
SUCCINATE.
GO TO PUBMED, ALPHA' KETOGLUTE
RATE IS MITOCHONDRIA, IRON IS
CONTROLLED BY MITOCHONDRIA,
OXYGEN CONTROLLED BY
MITOCHONDRIA.
SUCCINATE IS GENERATEDDED IN THE
MITOCHONDRIA.
YOU CAN SEE EARLY EVOLUTION ONE
WAY MITOCHONDRIA MIGHT HAVE
COMMUNICATED TO THE NUCLEUS OR
TO THE CYTOPLASM IS THANK YOU
THESE ENZYMES BECAUSE IT CAN
POTENTIALLY CONTROL THE
ACTIVITY, BY ALL A VARIETY OF
WAYS.
ONE THING THAT'S COME OUT OF
PEOPLE WHO STUDIED THESE ENZYMES
IS THE MOST POTENT REGULATOR IS
TWO HYDROXY GLUTE RATE.
SUCCINATE WILL ALSO INHIBIT
ENZYME NOT AS EFFECTIVE AS 2
HYDROXY GLUTE RATE.
2 AG AND ALPHA GLUTE RATE IS THE
SAME AND ONE IS OXIDIZED REDUCED
VERSIONS OF IT.
THE CANCER PEOPLE KNOW ABOUT
ALPHA KETOGLUTE RATE.
THEY'RE FOUND IN GLIOMAS AND AML
MAKE 2 HYDROXY GLUTE RATE.
MUCH UNDERSTANDING OF IT
FUNNELING INTO ENZYMES COMES
FROM THAT LITERATURE.
NOW, THE TYPE OF 2 HYDROXY
GLUTARATE THE IDEATIONS MAKE IS
THE R FORMTOR THE D FORM.
SO REMEMBER THIS PLURALITY.
THERE'S ANOTHER VERSION CALLED
THE L FORM OR THE S FORM THAT
EVERY PERSON IN THIS ROOM CAN
MAKE WITHOUT IDH MUTATION.
PLANTS MAKE IT.
IT'S BEEN CONSERVED THROUGHOUT
EVOLUTION. I WANT TO TALK ABOUT
HOW DO YOU MAKE NOT THE IDH
MUTATION VERSION OF 2 AG BUT THE
L.
I THINK THE IMPLICATION OF THIS
IS MUCH BROADER AND I WILL TELL
YOU WHY IN A SECOND.
SO THE WAY YOU MAKE L 2HE, OR
THE S 2AG, IS YOU ONLY MAKE IT
WHEN NADH LEVELS ARE HIGH.
NADH LEVELS ARE HIGH IF
RESPIRATORY CHAIN IS INHIBITED
OR IMPAIRED.
COMPLEX ONE'S JOB IS TAKE NADH
MAKE NAD SO MOVES US HAVE A LOT
MORE NAD THAN NADH IN
MITOCHONDRIA.
BUT IF WE'RE SEVERELY HYPOXIC OR
IF WE FAKE A POISON INHIBITS OUR
RESPIRATORY CHAIN OR AS MANY
ALLUDED DISEASES LIKE
PARKINSON'S OR NATURAL AGING YOU
SEE A DOWN REGULATION OF
RESPIRATORY CHIN AND YOU START
TO SEE A BUILD UP OF NADH
COMPARED TO NAD, THERE'S A WHOLE
LITERATURE ABOUT GIVING NKD
SUPPLEMENTS NOW.
I HAVE ASKED PEOPLE WHAT DOES
NAD DO?
THIS IS ONE EXPLANATION.
WHEN THE NADH LEVELS INCREASE
COMPARED TO NAD, MARKSAL
DEHYDROGENASES, IT TAKES
CONVERTS TOM ACETATE.
IT'S NATURALLY PREFERRED
SUBSTRATE BUT IT WILL
PROMISCUOUSLY UTILIZE ALPHA
KETOGLUTER RATE AND IF THERE'S A
LOT OF NADH AROUND THE MAL
DEHYDROGENASE WILL CONVERT TO 2
HYDROXY GLUTERATE.
YOU NEED THE NADH TO DRIVE THE
REACTION.
THERE'S AN ENZYME CALLED 2
HYDROXY DEHYDROGENASE THAT WILL
GIVE RID OF THE BUILD UP.
THE BRAIN HAS THE HIGHEST LEVELS
DEHYDROGENASE, PATIENTS WHICH
HAVE MUTATION IN THAT ENZYME
WILL ACCUMULATE 2 AG AND GET ALL
SORTS OF NEURAL DEVELOPMENTAL
AND NEURAL DEGENERATED
PHENOTYPES.
SO WE CAN SEE HOW 2 AG MIGHT BE
LINKED TO NEURODEGENERATION.
ONE THING IS THIS ENZYME IS
COMPLETELY DEPENDENT ON DUMPING
ELECTRONS TO COMPLEX 3.
SO WHEN WE KNOCK OUT COMPLEX 3,
AND WE FIRST DID IT IN OUR STEM
CELLS IN VIVO, WE SAW A LOT OF 2
AG ACCUMULATING, UP TO A
MILLIMOLAR.
AND IS BEING DRIVEN BUZZ NADH
BUILDS UP IN OUR COMPLEX 3 KNOCK
OUTS AND YOU CAN'T GET RID OF
IT.
THE OTHER PLACE IS LDH, WITH
ACIDIC PH CAN ALSO GENERATE 2 HG
SO GLYCOLYTIC CELLS.
YOU NEED NADH TO DRIVE THAT.
WE HAVE NOW HAVE MANY WAYS TO
RECONSTITUTE WITH ENZYMES AND
REPLENISH THE NAD POOL IN OUR
COMPLEX 3 KNOCK OUTS AND WE CAN
BRING DOWN 2 AG SO WE'RE
CONFIDENT THIS IS THE MECHANISM
BY WHICH YOU GENERATE THE 2 AG.
NAD MALI DEHYDROGENASE SYSTEM.
SO THE KEY IS WHETHER THE 2
HYDROXY GLOOM EAT RATE
ACCUMULATES IN THE TREGS.
AND WILD TYPES HAVE 50 TO 100
MICROMOLAR, ACCUMULATES UP TO
SOMEWHERE BETWEEN 3 TO 400
MICROMOLAR BUT THE KEY IS HOW TO
TRANSLATE TO ALPHA 2AG, IT HA Z
TO OUTCOMPETE AND WE START TO
SEE, DOING METABALOMICS OUT OF
THE MOUSE, IF A MOUSE DEES THREE
WEEKS BEFORE AND TAKE IT OUT AND
SHOOT THROUGH MASS SPEC.
THE SUCCINATE RATIO TO ALPHA KG
ALSO GOES UP AND SO THE QUESTION
IS, DOES THIS CORRELATE WITH
ANYTHING TO DO WITH DNA
METHYLATION.
TEN ENZYMES PARTICIPATE IN DNA
METHYLATION AND THEY HAVE BEEN
SHOWN IN VITROTOR SENSITIVE BOTH
TO ALPHA KG AND THE SUCCINATE.
THEY ARE ALPHA KG DEPENDENT IN
-- IF YOU PUT IT 2 AG OR
SUCCINATE YOU YOU CAN SEE
HYPERMETHYLATION OF DNA.
WE WENT BACK TO RAB SEQ AND SAID
LET'S LOOK AT THE -- RNA SEQ AND
LOOK AT 200 GENES DOWN REGULATED
AND DO BISULFITE DNA METHYLATION
SEQUENCING AND NOW AT THE TOP
DOWN REGULATING GENES, LOOK AT
THE CPG ISLANDS AS YOU SEE
HYPERMETHYLATION COMPARED TO THE
WILD TYPE.
SO THERE'S A CORRELATION BETWEEN
THE LOSS OF COMPLEX 3, LOSS OF
SUPPRESSIVE FUNCTION, DOWN
REGULATION OF KEY SUPPRESSIVE
GENES, HYPERMETHYLATION O DNA,
AND INCREASE IN THINGS LIKE 2
HYDROXY GLUTERATE AND SUCCINATE.
ALL CORRELATIVE.
THE REASON I KEEP ON SAYING, I'M
VERY CAREFUL ABOUT THIS, ONE OF
MY ORIGINAL DEGREE WAS IN MATH,
ONE THING YOU LEARN OVER AND
OVER IS CAUSALITY.
UNFORTUNATELY I DON'T HAVE A
GOOD CAUSAL EMPERIMENT.
THE CAUSAL EXPERIMENT WOULD BE
TO TAKE OUR COMPLEX 3 KNOCK
OUTS, AND FIND WAY TO GET RID OF
2 HYDROXY GLUTE RATE AND FIX THE
PHENOTYPE.
IF THAT'S THE DRIVER.
BEST WAY WOULD BE TO OVEREXPRESS
ENZYME.
WE CAN'T AND TRIED, THE PROBLEM
IS COMPLEX THREE KNOCK OUTS IF
WE HAVE ONE MAKE 2 HYDROXY
DILUTE RATE WE CAN HAVE THAT
SYSTEM OVEREXPRESS HYDROXY GLUTE
RATE BECAUSE COMPLEX 3 IS INTACT
WHEN COMPLEX 1 IS KNOCKED OUT.
SO WE'RE GOING BACK TO DO THIS
EXPERIMENT, ONE EXPERIMENT WE
CAN DO IS IN VITRO SUFFICIENCY,
WE CAN TAKE FOX P 3 NULL, WILD
TYPE RIGHT OUT OF A MOUSE, GIVE
IT ALL THE CYTOKINES KEEP THEM
HAPPY, GET THEM TO PROLIFERATE,
AND GIVE IT 50 MICROMOLAR 2
HYDROXY GLUTE RATE.
50 MICROMOLAR GIVES YOU 400
MICROMOLAR IN THE CELL.
IF YOU PUT 50 MICROMOLAR IN CELL
CULTURE AND BY METABALOMICS ASK
HOW MUCH THE CONCENTRATION IS
ENRICHED, IT'S ALMOST TENFOLD.
IT'S ABOUT WHAT WE SEE IN OUR
KNOCK OUTS.
SO WE'RE ON PAR, SO IN OTHER
WORDS YOU CAN ALWAYS THIS IS ONE
OF MIKE'S FAVORITE THINGS, YOU
CAN DUMP IN AS MUCH METABOLITE
OR TOXIN TO SEE EFFECT.
WE WERE CAREFUL TO PUT BACK THE
MONOHYDROXY GLUTERATE IN THE
WILD TYPE CELL WHETHER GIVING
THAT 2 AG IS SUFFICIENT TO DRIVE
DOWN REGULATION OF SOME KEY
GENES INCLUDING NEUROFILL LYNN
AND -- THE GENES THAT WE OBSERVE
WERE DOWN REGULATED BY CONTRAST
FOX P 3 NOT DOWN REGULATED,
REMAINS UNCHANGED SO THIS SORT
OF KEEPS US GOING DOWN THIS 2
HYDROXY GROUT RATE PATHWAY GOING
FORWARD.
BUT REALLY THE SIMPLE TAKE HOME
MESSAGE IS WHEN WE THINK ABOUT
WHY IN THIS CASE WHY ANY
MAMMALIAN CELL MISFIRES, TREGS
CAN SURVIVE WITHOUT HAVING
FUNCTIONAL RESPIRATORY CHAIN OR
FUNCTIONAL COMPLEX 3 DEFINE WAY
TO PROLIFERATE AND GROW, I FIND
FASCINATING, HOW ARE THEY IN
VIVO GETTING THEIR NUTRIENTS FOR
GROWTH.
BUT WHAT THEIR REALLY USING THE
RESPIRATORY CHAIN IS TO CONTROL
THEIR SUPPRESSIVE FUNCTION.
IN THE ABSENCE OF FUNCTIONAL
RESPIRATORY CHAIN, THEY DON'T
SUPPRESS PROPERLY.
THIS IS CONTINUES TO ADD TO THIS
IDEA THAT MITOCHONDRIA
COMMUNICATE WITH THE NUCLEUS.
WE WORKED ON THE IDEA THAT IT
RELEASE H 2O 2 TO FUNNEL TO
TRANSCRIPTIONAL NODES.
ONE OF THE CHALLENGES FOR THE
FIELD, THE THAT CONTINUES TO
HAPPEN IS EXACTLY HOW THE H 2 O2
SIGNAL TRANSDUCTION RELAY
HAPPENS IN THE PIONEERING
PROPOSING SOME ELEGANT MODELS
HOW THIS WORKS.
WE HAVE BEEN INTERESTED IN
TAKING THE HIF SYSTEM AND
FITTING WHAT 15 RESIDUES THAT
PEROXIDE WOULD OXIDIZE IN THE
HIF PATHWAY OR THE
PROTEOHYDROXYLACES THAT ACCOUNT
FOR THE BIOLOGY.
THE OTHER THING WE'RE EXCITED
ABOUT IS THIS IDEA THAT THE TCA
CYCLE METABOLITES COULD BUILD UP
TO CAUSE HAVOC.
SO IN OUR WORLD H 2 O2 IS
GENERALLY BENEFICIAL, SELECTED
UNDER PHYSIOLOGICAL CONDITIONS,
AND WHERE MITOCHONDRIAL
METABOLITES MIGHT BE TOXIC IS
WHEN YOU HAVE RESPIRATORY
IMPAIRMENT OR DOWN REGULATION OF
RESPIRATION OR SOMEHOW YOU HAVE
THE NAD NADH RATIO OF NOD
BALANCE AND TOO MUCH NAD THAT
MIGHT TRIGGER SOMETHING LIKE 2
HYDROXY GLUTERATE WHICH CAUSE
DNA METHYLATION HYPERMETHYLATION
BUT REMEMBER 2 HYDROXY GLUTERATE
BASED ON PATIENT DATA CAUSES
NEURODEGENERATION.
SO THAT'S SORT OF LEAVES US TO
THINK MANY OF THE NAD DEPENDENT
DECREASE IN NAD WHICH IS BEING
LINK TO PATHOLOGY, ONE OR TWO
HYDROXY GLUTERATE IS ONE ASPECT
OF THAT MISSING LINK.
AND IT IS A NEUROTOXIC MOLECULE
THAT CAUSES HAVOC.
BUT THE OTHER THING IS, WHETHER
NATURE ALSO ORIGINALLY SELECTED
THIS AS A PHYSIOLOGICAL SIGNAL
DURING THE DEVELOPMENT BUT ONLY
WHEN YOU ACCUMULATE HIGH LEVELS
IT BECOMES A PATHOLOGIC MOLECULE
JUST LIKE ROS CAN.
AGAIN, WE HAVE TO DEVELOP NEWER
MOUSE MODELS, TO PERTURB NAD
NADH RATIO THE TRIHYDROXY
GLUTERATE, MORE WAYS TO SLOP UP
COMPLEX 3, OTHER SOURCES TO
INVOKE BIOLOGY. ANYWAYS, THANKS
AGAIN, MIKE AND TO THE INSTITUTE
AND THANK YOU FOR YOUR PATIENTS.
I THINK I HAVE GOT ABOUT TEN OR
TWELVE MINUTES TO ANSWER SOME
QUESTIONS.
[APPLAUSE]
>> GREAT JOB.
THANK YOU VERY MUCH, FOR
QUESTIONS PLEASE COME UP TO THE
MICROPHONE.
YES PLEASE.
>> VERY NICE STORY PARTICULARLY
ABOUT THE EPIGENETICS PART BUT
I'M A LITTLE CONFUSED ABOUT YOUR
ASSERTIONS AND H 2 O2.
YOU HAVE SO MUCH SUPEROXIDE AND
CATALASE IN THE MITOCHONDRIA,
HOW DO YOU ACCOUNT FOR THIS?
>> EXCELLENT QUESTION.
HOW DOES SUPEROXIDE OR HYDROGEN
PEROXIDE EVER ESCAPE BECAUSE AS
WE KNOW THE MITOCHONDRIAL MATRIX
IS SUPER WELL BUFFERED WITH
ANTIOXIDANTS, CORRECT?
>> YEAH.
>> SO ONE OF THE REASONS WE
REALLY LIKE COMPLEX 3 AS A MAJOR
SITE FOR THE RELEASE OF
SUPEROXIDE, BECAUSE IT'S THE
ONLY COMPLEX OR THE ONLY SYSTEM
WITHIN THE MITOCHONDRIA THAT
GENERATES SUPER OXIDE THAT DUMPS
INTO THE MEMBRANE SPACE WHERE IT
LEAKS OUT AND THERE'S SOD 1 IN
THE MEMBRANE SPACE, WE CAN
CONVERT PEROXIDE AND BOOM YOU
GET IT RIGHT OUT OF THE
MITOCHONDRIA OR THAT SUPER OXIDE
GOES THROUGH VDAC CHANNELS.
WE HAVE SEEN THAT.
SO THAT'S ONE OF THE REASONS WE
LIKE COMPLEX 3 AS MAJOR SITE OF
THE SUPER OXIDE AND OBVIOUSLY BY
DUMPING IN MEMBRANE SPACE YOU
HAVE ACCESSIBILITY TO THE OUTER
MEMBRANE QUICKLY, WHERE WE WOULD
ARGUE THINGS LIKE HYDROXYLACES
ARE THE DIRECT TARGETS OF THE
MITOCHONDRIA WOULD HAVE TO BE.
ONE OF THE THINGS THAT SUPPORTS
THIS IDEA IS UNDER HYPOXIA,,
THIS IS THE WORK OF MARK
GILLESPIE, HE SHOWED WITH
SENSORS AND EVERYTHING SOON AS
HE MADE CELLS HYPOXIC.
YOU HAVE DIFFUSE MITOCHONDRIAL
STAINING PATTERN, THEY ALL WENT
PERINUCLEAR.
AND HE COULD SHOW THAT THE
MITOCHONDRIA WERE THEN DUMPING
HYDROGEN PEROXIDE IN THAT
VICINITY.
THAT WAS ALL DUE TO COMPLEX 3.
>> DID YOU LOOK AT THE
GLUTATHIONE AND OTHER LABELS IN
THESE CELLS?
>> YEAH.
>> A LOT OF GLUTE TATHIONE AND
EVERYTHING ELSE.
JUST WONDERING IF THERE IS --
THERE'S SO MUCH OUT THERE.
>> YEAH.
SO THIS IS A MORE -- BEYOND
MITOCHONDRIA, AND AGAIN I SHOULD
DEFER TO -- AND HIS WORK ON
THIS, ONE OF THE QUESTIONS IS,
OKAY WE PROVIDE A SIMPLE
EXPLANATION HOW COMPLEX 3
RELEASE ROS IN THE RIGHT
COMPARTMENT IS THAT IS DUMPING
IT AND THE NEXT QUESTION, HOW
DOES THAT H 2 O2 RELAY ITS
SIGNAL TO EVENTUALLY CAUSE
CYSTEINE MODIFICATION WHICH ALER
PROTEINS. THERE'S A VARIETY OF
MODELS THAT HAVE BEEN PROPOSED
INCLUDING THE PROXY THEMSELVES
RELAYING, ANOTHER MODEL CALLED
THE FLOOD GATE MODEL THAT AGAIN
THIS IS A WORK OF SUBARY AND
COLLEAGUES.
SO THESE THINGS CONTINUE TO BE
TRIGGERED OUT AND THOSE ARE
STILL PERTINENT QUESTIONS.
>>
>> ONE MORE QUICK QUESTION.
THAT IS DID YOU LOOK AT PHOTO
RECEPTORS IN YOUR NICE?
BECAUSE PHOTO RECEPTORS USE
MAXIMUM AMOUNT OF OXYGEN FOR ANY
POST -- CELL.
>> NO.
IF YOU'RE INTERESTED WE'RE HAPPY
TO SEND YOU THE MICE.
GREAT.
CROSS IT TO A CRE AND SEE WHAT
YOU GET.
>> THANK YOU VERY MUCH.
I'M GLAD AFTER TWO DECADES OF
STRUGGLING TO INTRODUCE THE ROLE
OF IMMUNITY IN CANCER PEOPLE ARE
TALKING ABOUT THESE FANTASTIC
TOPICS IN CANCER DEVELOPMENT.
I THINK WHAT YOU WERE MENTIONING
ABOUT MITOCHONDRIA GOING OFF AND
ON AND BEING IN WAY TO CANCER,
WE HAVE DETAILED THIS WHAT
HAPPENS WHEN YOU INDUCE GROWTH
PROMOTING ROLE OF TUMOR GENIC
PROPERTY OF ACUTE INFLAMMATION
YOU SHUT DOWN THE MITOCHONDRIA.
SO THAT THE MITOCHONDRIA HAS
CHANGED TO REGENERATE SUCCINATE
AND ALL THOSE OTHER MEDIATORS
THAT YOU MENTION.
IN THIS PROCESS THE SHUTTLE TO
MITOCHONDRIA ALSO IS AFFECTED.
MY QUESTION IS, WHEN YOU WERE
TALKING ABOUT ALPHA
KETOGLUTERATE OR LDH, HAVE YOU
LOOKED AT SOME MULTI-METABOLISM
OF THE MACRO MOLECULES THAT ARE
ALSO NEEDED, THE ENERGY FOR
METABOLIZING ESSENTIAL AMINO
ACIDS ISOLEUCINE.
>>
>> RIGHT.
>> HAVE YOU LOOKED AT THOSE?
AND ALSO MANY OTHER COMPONENTS,
THIS IS VERY INTERESTING TOPIC.
>> THANK YOU FOR BRINGING UP.
WE HAVE NOT BUT THE COMMUNITIES
ARE VERY INTERESTED IN BRANCHING
APPLY KNOW ACIDS -- AMINO ACIDS.
ONE OF THE PLACES THEY LIKE TO
DUMP THE CARBONSES IS ONE TO
ACETYL COA LEUCINE AND OTHER ONE
VALINE UNDER SUCCINIL COA, THIS
CAN FULFILL THE TCA CYCLE.
THERE'S A SIMPLISTIC VIEW THAT
EVERYTHING IS ABOUT GLUCOSE TO
ACETYL COA TO ALPHA
KETOGLUTERATE BUT ONE OF THE
ONES WHICH CAN DO IT WE FIND AT
LEAST IN CELL CULTURE THAT CAN
SUBSTITUTES VERY WELL IS
BRANCHING AMINO ACIDS.
BRANCHING AMINO ACIDS ARE
ELEVATED EARLY LIKE PANCREATIC
CANCER SO I ARGUED THAT THERE'S
GOOD REASONS TO THINK ABOUT
BRANCHING AMINO ACID METABOLISM
AS SORT OF AN ANOPOROTIC
FULFILLING THE TCA CYCLE
METABOLITES.
BUT WE HAVEN'T OURSELVES
MANIPULATED THIS SYSTEM.
I KNOW THERE ARE OTHERS
INTERESTED IN DOING THAT.
>> IT'S A FASCINATING TOPIC.
I HAVE PUBLISHED IT RECENTLY, IF
YOU'RE INTERESTED WE CAN
DISCUSS.
>>
>> THAT WOULD BE GREAT.
THANK YOU.
>> THANK YOU, ON OUR LEFT.
>> SO IN THE TREG SETUP OR BACK
GROUND, HAVE YOU TRIED
EXPRESSING THE ALTERNATIVE
OXIDASE?
BECAUSE IN THAT CASE 2 HYDROXY
GLUTERATE LEVELS GO DOWN BECAUSE
YOU -- THE ALTERNATIVE DOESN'T
GENERATE ROS LIKE -- SO WHAT
HAPPENS?
>> EXCELLENT.
SO LET ME JUST -- SO AS I SHOWED
YOU THE WAY WE RESCUE THE
TUMORIGENITICITY IN THE CANCER
WAS WE KNOCKED OUT COMPLEX 3,
THE TUMORS DIDN'T GROW, WE PUT
THIS ANCIENT ENZYME ALTERNATIVE
OXIDASE WHICH ALLOW COMPLEX 1
AND 2 TO WORK IN ABSENCE OF 3
AND IT RESCUED THE TUMORS, ALLOW
IT IS METABOLITES TO COME BACK
AND THE 2 AG DOES GO DOWN IN
THAT SETTING.
WE HAVE DONE THOSE EXPERIMENTS.
WE SHOULD BE ABLE TO USE THE
SAME STRATEGY, IN OUR TREG SO WE
GENERATE AD CONDITIONAL LOCK
STOP LOX AND THE LOCUST OF THE
AOX AND WE'RE NOW CROSSING THAT
TO OUR COMPLEX 3.
THAT'S EXACTLY -- WOULD BE ONE
OF THE WAYS.
AND WHAT'S NICE AS YOU POINTED
OUT, IT DOESN'T RESCUE THE ROS.
ALL IT RESCUES THE METABOLITES.
>> THANKS.
>> ON OUR RIGHT.
>> VERY NICE.
SO THE ONCOGENESIS MODEL YOU
CHOSE THE RAS DRIVEN MODEL RAS
DRIVEN ONCOGENESIS HAS BEEN
SHOWN TO DEPEND IN SOME PART ON
NF KAPPA B TO PREVENT RAS
INDUCED SENESCENCE.
SO DID YOU LOOK TO SEE WHETHER
THIS MIGHT BE DEPENDENT ON NF
KAPPA B AND ALSO IF YOU USE
NON-RAS DRIVEN MODEL DO YOU SEE
A SIMILAR DEPENDENCE IN TUMOR
JOE AT THIS IN THISTY?
>> WE HAVE NOT NF KAPPA B.
I THINK WE HAVE BEEN LITTLE
NAIVE IN SOME EXPERIMENTS, A
LITTLE OBSESSED BY SHOWING THE
NECESSITY OF THE RESPIRATORY
CHAIN AND COMPLIMENTING WITH ALL
THESE ANCIENT ENZYMES TO FIGURE
EXACTLY WHAT ASPECT OF THE
RESPIRATORY CHAIN BUT AS WE GET
CLOSER NO U THE NEXT STEP, OKAY,
THEN WHAT?
THAT'S FOR US BOTH NF KAPPA B AN
HIF CONTINUE TO BE THE ONES
WE'RE MOST INTRIGUED BY.
YOUR SECOND QUESTION WAS ABOUT
--
>> JUST WHETHER --
>> WHETHER OTHER MODELS.
>> A NON-RAS MODEL --
>> RIGHT.
SO WE HAVE DONE A MIG MODEL,
OSTEOSARCOMA INDUCIBLE MIG MODEL
THAT DEAN FILCHER'S LAB MADE
SEVERAL YEARS AGO AND THE
RESULTS HOLD UP IN THOSE MODELS.
I THINK MORE AND MORE THERE'S
PEOPLE USED A VARIETY OF GENETIC
AND PHARMACOLOGICAL WAYS OF
SUPPRESSING THE RESPIRATORY
CHAIN AND THEIR FINDING SIMILAR
THINGS.
PROBABLY THE ONE THAT I'M MOST
EXCITED ABOUT IS THAT CELLS THAT
EMERGE, THAT ARE RESISTANT TO A
PARTICULAR THERAPY,
ANTI-ANGIOGENIC THERAPY,
CHEMOTHERAPY, BRAF OR SOME
TARGETED THERAPIES, THE CELLS
THAT PERSIST FOR A WHILE, AND
THEY'RE SLOW GROWING AND MIGHT
HAVE SO CALLED TUMOR INITIATING
STEM LIKE -- THOSE CELLS ALL
TAUGHT ARE MUCH MORE SENSITIVE
TO MITOCHONDRIAL RESPIRATORY
CHAIN INHIBITORS THAN THE
ORIGINAL TUMORS.
I THINK THAT'S QUITE EXCITING.
AVENUE TO PURSUE.
>> EMERGING AREA.
SO WE'RE GOING TO HAVE A
RECEPTION IN THE NIH LIBRARY
AFTER THIS LECTURE BUT LET'S
THANK NAV AGAIN FOR GIVING US A
GREAT PRESENTATION.
[APPLAUSE]
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