HE RECEIVED THE ARTHUR S.
FLEMMING AWARD FOR OUTSTANDING
GOVERNMENT SERVICE AND IN 1987
THE YEAR OF HIS UNTIMELY DEATH
AT AGE 43 HE WAS ELECTED TO THE
NATIONAL AKD--SALLY ME OF
SCIENCES.
YOU CAN IT TELL I'M A MAC
PERSON.
AND THIS IS A PHOTO OF GEORGE
KHOURY ON YOUR LEFT WITH A VERY
YOUNG PETER HOWLY AND MALCOM
MARTIN WHO WAS MY OWN Ph.D.
MENTOR -- POST DOCTORAL MENTOR
AND IS HERE IN THE AUDIENCE.
SO DURING HIS SHORT CAREER
GEORGE PUBLISHED OVER 140 PAPERS
DECIPHERING MECHANISMS OF
EUKARYOTIC TRANSCRIPTION.
MANY OF THESE USE VIRUSES AS
MODEL SYSTEMS AND WE HAVE A MAC
TO PC GASHLEING THERE.
GEORGE HAD VERY DISTINGUISHED
TRAINEES AND COLLABORATORS
LISTED HERE.
AND SIMILARLY THE GEORGE KHOURY
MEMORIAL LECTURES HAVE BEEN
GIVEN BY A VERY DISTINGUISHED
GROUP OF SCIENTISTS MOST
RECENTLY JULIE 5ER, SUSAN WEISS,
AND THIS BRINGS US TO TODAY'S
SPEAKER, FOR ILEANA CRISTEA, SHE
DID HER HONOR GRADUATE AND
Ph.D. WORK AT UNIVERSITY OF
MAN THE CHESTER IN THE UK --
MANCHESTER IN THE UK, IN
PROTEOMICS AND TOXICOLOGY, DID A
POST DOC AT ROCKEFELLER
UNIVERSITY BEFORE BEING
RECRUITED TO PRINCETON
UNIVERSITY WHERE SHE ROSE REALLY
REMARKABLY QUICKLY THROUGH THE
RANKS TO BECOME FULL PROFESSOR
IN 2016.
ILEANA HAS RECEIVED NUMEROUS
AWARDS, TOO MANY TO LIST MERE
BUT THESE INCLUDE AN AVANT GUARD
PILOT PROJECT OR NEAR AWARD,
DISCOVERY AWARD IN PROTEOMIC
SCIENCES, DIRECTOR OF THE
GRADUATE STUDIES AT PRINCETON,
GRADUATE AWARD AT PRINCETON AND
AS OF LAST YEAR, MOLECULAR
CELLULAR PROTEOMICS.
ILEANA MADE REALLY MAJOR
CONTRIBUTIONS TO SEVERAL AREAS
OF RESEARCH INCLUDING DEVELOPING
PROTEOMIC METHODS FOR DISTURBING
VIRUS INTERACTIONS, NUCLEAR
SENSING OF VIRAL DNA,
EVOLUTIONARY CONSERVED ANTIVIRAL
FACTORS AND REGULATION OF
PROTEIN ACETYLATION.
SO I WILL LIKE TO WELCOME ILEANA
TO THE PODIUM FOR HER LECTURE,
THE INTERFACE BETWEEN METABOLISM
AND IMMUNITY WITHIN A VIRUS
MICROENVIRONMENT, WELCOME.
[ APPLAUSE ]
>> THANK YOU SO MUCH FOR THAT
VERY KIND INTRODUCTION, IT IS
JUST SUCH A GREAT HONOR BEING
HERE AND SEEING OLD FRIENDS AND
NEW FRIENDS AND JUST REALLY, I
CANNOT TELL YOU HOW HONORED I AM
TO BE INVITED TO GIVE DR. GEORGE
KHOURY MEMORY MEMORIAL LECTUREI
WANT TO SAY A FEW WORDS ABOUT
THIS.
I THINK ERIC ALREADY TOUCH
OFFICE OF DIVERSITY THIS IN
TERMS OF FUNDAMENT AMILLIO
SCIENTIFIC DISCOVERIES THAT
GEORGE CONTRIBUTED TO THE FIELD
OF MOLECULAR BIOLOGY, HIS WORK
ON ENHANCERS AND ALSO ON
UNDERSTANDING TRANSCRIPTIONAL
CONTROL OF GENE EXPRESSION HAS
BEEN EXTRAORDINARY BUT I WANT TO
SAY ALSO FROM THE PERSPECTIVE OF
PRINCETON, ERIC TOLD YOU THAT
GEORGE ACTUALLY HAD HIS
UNDERGRADUATE DEGREE AT
PRINCETON AND I JUST WANT TO SAY
THAT HE BECAME SUCH A STRONG
SUPPORTER OF PRINCETON
UNIVERSITY AND SO, COMMITTED TO
RECOGNIZING PEOPLE WHO HAVE
CONTRIBUTED TO TRAINING HIM AND
HE HAS BEEN IN CONSTANT KIND OF
TOUCH WITH PRINCETON UNIVERSITY
BECOMING AN ADVISOR TO THE
PRINCETON UNIVERSITY PRESIDENT
AND REALLY HELPING TREMENDOUSLY
TO CREATE THE DEPARTMENT OF
MOLECULAR BIOLOGY AND TO RECRUIT
A NUMBER OF FACULTY AS KIND OF
THE INAUGURAL FACULTY TO START
THIS NEW DEPARTMENT OF MOLECULAR
BIOLOGY FOR PRINCETON AMONGT
THESE ARE DR. LAVINE AND TOM
SHANK, A WONDERFUL VIROLOGIST,
WHO I GOT TO COLLABORATE WITH AT
THE UNIVERSITY WHEN I WAS A POST
DOC AND HE ENCOURAGED ME TO
APPLY AT PRINCETON TO APPLY FOR
A FACULTY POSITION AND I WANT TO
SAY GEORGE HAS IMPACTED MY
CAREER IN ANOTHER WAY EVEN
EARLIER, IN THE 1970S, WHEN HERE
AT THE NIH HE IRPT ACTED WITH
DR. LYNN QUIOF THE AND AT THE
TIME, LYNN WAS CLOSE TO HIS
LABORATORY AND LYNN CREDITS
GEORGE FOR ENCOURAGING HIM AND
SUGGESTING THAT ME MAYBE SHOULD
CHANGE -- USE HIS SKILLS TO
CHANGE SOME OF HIS SCIENTIFIC
FOCUS FROM A COURAGEOUS WAY FROM
BACTERIAL PHAGES AND LUNG PHAGES
TO EUKARYOTIC VIRUSES AND LYNN
LATER ON JOINED PRINCETON
BECOMING THE CHAIR OF THE
DEPARTMENT OF MOLECULAR BIOLOGY
AND BEING THE 1 WHO ENDED UP
RECRUITING ME AND HIRING ME AT
PRINCETON.
SO I MAY NOT BE HERE SPEAKING TO
YOU WITHOUT GEORGE'S
CONTRIBUTION TO MOLEC BAR
BIOLOGY AND TO SPRINS TON. --
PRINCETON.
SO THANK YOU SO MUCH FOR HAVING
ME HERE.
A LITTLE BIT ABOUT MY TALK TODAY
AS YOU HEARD ISSUES I WOULD LO
OF TO TELL YOU ABOUT HOW SOME
RESEARCH DIRECTIONS IN MY LAB
HAVE EVOLVED TO LOOK INTO THIS
INTERFACE BETWEEN METABOLISM AND
IMPACT ON IMMUNE SIGNALING IN
THIS CONTEXT OF VIRUS
MICROENVIRONMENT AND FIRST I
WILL SAY WHAT MY DISCLOSURES
ARE.
I'M A CONFOUNDER AND MEMBER OF
THIS SCIENTIFIC ADVISORY BOARD
FOR ADVERSE BIOTHAT FOCUSES ON
ANTIVIRAL TREATMENT, TARGETING
CITUANS, AND ALSO DEVICES THAT
ARE FOCUSING ON METHODS FOR
SEPARATING PROTEINS AND OF
COURSE ANALYSIS WITH MASS
SPECTROMETRY, BUT WHEY WILL TELL
YOU IS NOT CONNECTED IN ANY
SHAPE OR FORM WITH THESE 2
POSITIONS THAT I HAVE HELD OR
HOLD.
OKAY, SO, THE STORY THAT I WANT
TO TELL YOU AGAIN AND TO PUT IT
IN CONTEXT STARTED FROM OUR
INTEREST IN UNDERSTANDING
DIFFERENT TYPES OF CELLULAR
COMMUNICATION DURING VIRAL
INFECTION AND OF COURSE THERE
ARE MANY DIFFERENT TYPES OF
COMMUNICATION DURING INFECTION
IN A HOST CELL.
SO AT AN INTRACELLULAR LEVEL, WE
KNOW AN INFECTION TRIGGERS
CHANGES IN THE SE, CHANGES NOT
ONLY THE COMPOSITION OF A CELL
BUT THE ORGANELLE OF THE CELL,
AND THEY MAY LOOK DIFFERENTLY.
AND I WANT TO SPEND THE FIRST
PART OF MY TALK LOOKING AT HOW
WE LOOKED AT INTRACELLULAR
COMMUNICATION THAT IS
FACILITATED BY ORGANIZATION
CHANGES, AND BY CHANGES IN
ORGANELLE-ORGANELLE CONTACTS AND
WHAT THIS MEANS.
AND THEN, I WILL TRANSITION TO
CONSIDERING THIS INFECTED CELL
AS PART OF ITS ENVIRONMENT, AS
PART OF ITS SYSTEM AND LOOKING
AT HOW THIS INFECTED CELL CORN
VEYS THIS INFORMATION AND HOW IT
AFFECTS ITS ENVIRONMENT, THE
CELLS THAT ARE IN THE IMMEDIATE
PROXIMITY OR THOSE THAT ARE
FURTHER AWAY AND NOW WE HAVE
TRIED TO KIND OF CHARACTERIZE
THIS, STILL NOT SUFFICIENTLY
WELL CHARTED ENVIRONMENT OF THE
VIRUS MICROENVIRONMENT.
AND I WILL TRY AS I LOOK AT THIS
INTRACELLULAR AND INTRACELLULAR
STORIES THAT I WILL TELL YOU
ABOUT, WHAT THE RELEVANCE IS IN
TERMS OF INFECTION AND CO
INFECTIONS AND DISEASE AND
INFECTION IN THESE PATHOLOGIES,
SO TO START THE INTRACELLULAR
LEVEL, AS I TOLD YOU VIRUSES --
INFECTIONS WITH VIRUSES INDUCE
SUCH PROFOUND CHANGES IN
ORGANIZATION OF THE CELL AND
STILL UNDEREXPLORED AREA OF
VIROLOGY IS THAT ORGAN
REMODELING AND THAT IS SO
FUNDAMENTAL TO I HAVE USS YOU
CAN THINK OF, THAT VIRUSES NEED
TO LET ME SEE IF THIS WORKS.
OH, I'M TRYING TO -- OKAY, I'M
LOOKING, IF I HAVE A POINTER,
MAYBE I WILL JUST USE THIS.
CAN YOU SEE MY POINTER?
NO.
OKAY.
IESM GETTING VERY QUICK
TECHNICAL SUPPORT, I SEE.
NO, DOESN'T WORK.
IT'S ALL RIGHT.
I SEE A POINTER ON YOUR
PRESENTATION, THAT APPEARS, SO
SOMEBODY WAS TRYING TO HELP YOU
TO POINT.
OKAY, SO, VIRUSES INDUCE THIS
NUMEROUS CHANGES IN ORGANELLE
SHAPE AND COMPOSITION AND THAT
CAN BE PUT UNDER THE UMBRELLA OF
ORGANELLE REMODELING AND SHE'S
ARE SO IMPORTANT FOR ENTRY OF A
VIRUS INTO HOST CELLS FOR THE
REPLICATION OF THAT VIRUS,
EITHER IF IT'S NUCLEAR OR
CYTOPLASMIC FOR THE ASSEMBLY OF
NEW INFECTIOUS VIRAL PARTICLES
OR FOR THE EGRESS, AND I'M
SHOWING THIS HERE FOR BETTER
HERPES VIRUS, IS REALLY AN
IMPORTANT VIRUS, WIDELY SPREAD
AND CONNECTED TO LOTS OF
DIFFERENT CHRONIC DISEASE,
INCLUDING CARDIAC DISEASE BUT
ALSO THE LEADING CAUSE OF VIRUS
INNED BIRTH DEFECTS BUT WE CAN
LOOK AT MANY OTHER VIRUSES AND
WE WILL FIND SIMILARLY,
DIFFERENT RANGE OF ORGANELLE
REMODELING EVENTS, SO FOR
EXAMPLE, ATMV, THAT I TOLD YOU
EARLIER, INDUCES THIS CHANGES IN
NUCLEAR PERIPHERY AND THE
ASSEMBLY COMPLEX YOU SEE ON THE
RIGHT HAND SIDE, HSV1 ALSO
TARGETS THE NUCLEOTIDES CLER
PERIPHERY, IT ALSO FRAGMENTS
MITOCHONDRIA, INFLUENZA A ALSO
REMODELS MITOCHONDRIA.
CORONA VIRUSES ARE INDUCING THIS
DOUBLE MEMBRANE VESICLES FORMED
OF ER, THAT ARE REALLY IMPORTANT
FOR THE REPLICATION OF THIS
VIRUS, SO ALL THESE CHANGES IN
THE SHAPE AND THE MORPHOLOGY OF
ORGANELLES THAT ARE CAREERICALLY
CONTROLLING AND REGULATING THE
ORGANELLE FUNCTION THAT HAPPENED
DURING INFECTION THAT ARE NEEDED
FOR INFECTION TO PROGRESS.
THIS IS AS I SAID STILL AN
UNDEREXPLORED AREA OF VIROLOGY,
SO FOR MANY OF THESE EVENTS WE
DO NOT UNDERSTAND EXACTLY WHAT
THE FUNCTIONS ARE, OR IF THEY'RE
CONNECT INDEED ANY WAY TO SOME
OF THE LINK, INFECTION INDUCED
PATHOLOGYINGS, ALSO, HOW IT IS
COORDINATED.
OF COURSE IF A SHAPE IS
DIFFERENT, IT WILL AFFECT
IMMEDIATELY ITS ENVIRONMENT,
LIKE IF I SPREAD MY ARMS OR IF
I'M ALTOGETHER, I WILL IMPACT
DIFFERENTLY AND MAKE DIFFERENT
CONNECTIONS WITH MY IMMEDIATE
ENVIRONMENT IN THE CELL.
AT THE SAME TIME HOW ARE THEY
COORDINATED IN SPACE AND TIME.
OR FUNCTIONALLY COORDINATED, SO
WE HAVE INVESTIGATED THIS FOR A
NUMBER OF YEARS IN MY LAB AND I
WANT TO GIVE 1 EXAMPLE FROM SOME
STUDIES THAT WE HAVE DONE SOME
YEARS AGO THAT REALLY HAVE
STARTED TO PUSH US MORE AND MORE
TOWARDS THIS IDEA OF
COORDINATION.
THAT THIS ORGANELLE REMODELING
EVENTS ARE ACTUALLY LINKED TO
EACH OTHER AND THEY ARE
AFFECTING EACH OTHER AND THIS
WAS FROM WORK WE HAVE DONE ON
PEROX DOME WHERE WE WERE
STUDYING THE MEG LA LO VIRUS AND
HSV1, AND WE TART TO SEPARATE
ORGANELLES BY COMPOSITION AND
FIND OUT HOW THEY CHANGE AND
THEN WE COMBINE THIS WITH
STRUCTURAL MICROSCOPY STUDIES
AND ALSO WITH EMILY EDUCATIONAL
ORDER OF MICRONSIC STUDIES AND
MATHEMATICAL MODELS AND MACHINE
LEARNING TO START TO LEARN AND
ASSIGN COMPONENTS, VIRAL HOST
COMPONENTS TO DIFFERENT
ORGANELLES OR UNDERSTAND WHAT
DOES IT MEAN IN TERMS OF THE
STRUCTURE FUNCTION RELATIONSHIPS
AND WHAT WE FOUND IS THAT
PEROXISOMES, I AM STILL HOPING I
CAN GET A POINTER HERE.
LET ME SEE IF I CAN DO THIS.
>> IT'S NOT WORK?
>> NO, IT'S NOT WORKING BUT
MAYBE I NEED TO -- YES.
YEAH, IT'S ALL RIGHT.
SO IN THIS CIRCLE HERE, CAN YOU
SEE PEROXISOMES, ARE CHANGING
IN SHAPE AND NUMBER AND THAT
THESE CHANGE IN MORPHOLOGY
ACTIVATING ENZYMES THAT ARE
RESPONSIBLE FOR INITIATING LIPID
SYNTHESIS, THE AND WE SHOWED HOW
THESE PLACENTAS MALLOW
GENERATEDS AND BECOME INFECTIOUS
PARTICLES SO THE REASON I'M
SAYING COORDINATION IS BECAUSE
THE LIMIT SYNTHESIS STARTS IN
THE PEROXISOME, BUT IT'S BASED
-- THE LIPIDS HAVE TO BE MATURED
IN THE E. R. SO THIS IS BASED ON
THE INTERACTION OF PEROXISOME
AND THE CONTACTS.
SO THE MORE WE INTERROGATE THE
REMODELING EVENTS, THE MORE WE
WERE STRUCK ON HOW THIS IS AND
THE CONTACT, WE DO ALL KNOW THAT
THE THEY HAVE SEPARATION BUT
RATHER THEY FORM THIS INTRICATE
NETWORK OF COMMUNICATION INSIDE
THE CELL THAT ARE SO IMPORTANT
FOR SO MANY FUNDAMENTAL CELLULAR
PROCESSES, AND THESE
INTERACTIONS ARE FACILITATED BY
PROTEINS CALLED MEMBRANE CONTACT
SITES.
I SEE SOME PROMISE.
>> YOU HAVE TO ACTIVATE IT ON
HERE.
IT MIGHT BE THIS LITTLE BUTTON.
>> NO WORRY.
NO WORRIES.
>> AND THEN --
>> VICTORY.
>> OTHERWISE IT WAS MY POINTING,
I WOULD LOVE TO SEE THAT CHRIS,
I WOULD HOPE FOR THE SECOND
OPTION.
ALL RIGHT, SO THIS INTERACTIONS
BETWEEN ORGANELLES ARE
FACILITATED BY THESE PROTEINS
CALLED MEMBRANE SITES THAT ARE
BRINGING THEM IN CLOSE PROXIMITY
SO THESE ARE NOT FUSION EVENTS
AND THEY CAN FACILITATE DIRECT
TRANSFER OF MOLECULES LIKE
LIPIDS AND CALCIUM AND I'M
SHOWING YOU HERE 1 OF OUR
MICROSCOPY IMAGING WHERE'VE SEEN
RED PEROXISOMES IN THE ER AND
YOU SEE THE DYNAMIC INTERACTION
IS AND I WAS TELLING YOU ON THE
PREVIOUS SLIDE ABOUT THIS ER,
PEROXISOME CONTACT.
SO WE WONDERED WHETHER A LOT OF
THESE ORGANELLE REMODELING
COMPLEXES, OR A LOT OF THESE
AGER ANLE REMODELING EVENTS THAT
I TOLD YOU ABOUT ARE UNDERLYING
SOME OF THIS -- ARE BASED SORRY
ON CHANGES IN MEMBRANE CONTEXT
SIDE AND WHETHER THESE PROTEINS
THAT FORM MEMBRANE CONTACT SITES
ARE FACILITATING OR DRIVING OR
REGULATED BY THIS ORGANELLE
REMODELING EVENTS SO LOOKING AT
THIS DURING INFECTION AND WE HAD
FIRST DEVELOPED A METHOD THAT
WOULD ALLOW US TO DETECT THIS
HARD TO DETECT MEMBRANE CONTEXT
PROTEIN, SOME OF THEM ARE
MEMBRANE BOUND, WE DIDN'T HAVE
ANTIBODIES FOR A LOT OF THIS, SO
CAITLIN IN MY LAB TARGETED THIS
ASSAY THAT ALLOW HER TO DETECT
ALL THE KNOWN MEMBRANE CONTEXT
SIDE PROTEINS, DETECT THEM AND
QUANTIFY THEM AND THEN APPLY
THIS TO THE 4 DIFFERENT VIRUSES
THAT I MENTIONED EARLIER, THIS D
MA VIRUS, AND INFLUENCE AND THIS
BETA CORONA VIRUS, AND LOOK THEA
HOW THESE CONTACTS ARE CHANGING
AS THE INFECTION PROGRESSES AND
THIS KIND OF INITIATED SOME
REALLY INTERESTING RESEARCH
DIRECTIONS IN OUR LAB AND I WANT
TO TELL YOU WHAT WE HAVE LEARNED
FROM HER FINDINGS ON OTHER
MITOCHONDRIA ER CONTACTS
ESPECIALLY DURING HCMV, SO IF
YOU THINK ABOUT MITOCHONDRIA, WE
KNOW THAT MITOCHONDRIA ARE SUCH
A REGULATORY HUB DURING VIRAL
INFECTION, SO MANY VIRUSES ARE
TARGETING MITOCHONDRIA EITHER
INDUCING FUSION OR FRAGMENTATION
OR FISSION, AND THIS IS IN ORDER
TO CONTROL RES PERRATION, SO
ENERGY PRODUCTION, IMMUNE
SIGNALING OR APOPTOSIS.
AND USUALLY WHAT WE KNOW IS THAT
WHEN MITOCHONDRIA ARE BEING
FRAGMENTED THAT'S CONNECTED TO A
DECREASE IN RESPIRATION,
CONNECTED TO DECREASE BIOGENIC
OUTPUT AND HERE YOU SLEEP APNEA
AND OBESITYY SOME VIRUSES THAT
EITHER INDUCE FRAGMENTATION, BUT
SOMETHING'S BEEN PUZZLING IN THE
FIELD OF HMV VIROLOGY IS THAT IT
INDUCES STRIKING FRAGMENTATION
OF MITOCHONDRIA AS YOU SEE HERE.
BUT THIS MITOCHONDRIA IS ALSO
BIOGENETICALLY ENHANCED SO WE
SEE INCREASE IN PRODUCTION.
SO CAITLIN DISCOVERED AT THE
TIME WHEN THIS PHOSPHORYLATION
OCCURS THERE IS ALSO INCREASED
CONTACT BETWEEN MITOCHONDRIA AND
THE ER, THROUGH THOSE MEMBRANE
CONTACT SITE PROTEINS THAT
FACILITATE THE CONTACT, SHE
DELIVER INVESTIGATED THIS
CONTURNING TO MIRROR IMAGE
COSCOPY AND WHAT SHE FOUND IS
THAT THIS CONTACT IS REALLY
CHANGED DURING INFECTION, SO IN
UNINFECTED CELLS AND THESE ARE
PRIMARY HUMAN FIBROBLASTS, SHE
NOTICED THE USUAL AND KNOWN AND
TYPE OF INTERACTION BETWEEN
MITOCHONDRIA AND THE ER WHERE
THERE IS THIS DYNAMIC
INTERACTION, YOU SEE
MITOCHONDRIA IN RED AND THE ER
KIND OF WEAVING,A ROUND THE
MITOCHONDRIA AND HAVING THIS
DYNAMIC INTERACTION, BUT AFTER
INFECTION WITH ACMV, AND AS THE
INFECTION PROGRESSED, THE
FRAGMENTED MITOCHONDRIA BECAME
INCAPSULATED AND STABILIZED IN
ER POCKETS.
SO SHE TURNED THIS NEW SORT OF
CONTACT STRUCTURE MENC AND WENT
AND, HADED AND CONFIRMED THIS
USING USING CRYO MICROSCOPY AND
YOU SEE HERE HOW IT'S SURROUNDED
BY THE ER AND THEN WE
INVESTIGATE TD ITS FUNCTION.
WE FOUND WHAT PROTEINS ARE
NECESSARY FOR THE FORMATION OF
MENC, AND WE SHOW THAT SOMEHOW
THE STRUCTURE FUNCTIONS SOMEHOW
FOR THE BENEFIT OF THE VIRUS AND
SINCE THEN WE INVESTIGATED HOW
THIS HAPPENS AND WHY AND THIS
WAS WORK DONE BY WILL HOFSTAT A
Ph.D. STUDENT IN MY LAB.
IN OTHER WORDS WHY ARE THEY
FORMED AND WHY ARE THEY
FUNCTIONING FOR THE BENEFIT OF
THE VIRUS AND WILL WAS ABLE TO
THOUGH IS FIRST LOOKING AT HOW
MENCs ARE FORMING HE
CONSIDERED 2 THINGS HE
CONSIDERED HOW IT'S FRAGMENTED
AND DURING INFECTION AND HE
FOUND THAT ATMV SUPPRESSES
FUSION OF MITOCHONDRIA BUT
PROMOTES FRAGMENTATION THROUGH
THIS PERIPHERAL FISSION, INSTEAD
OF THE CLATIONZIC MIDZONE
VISION, IT'S THE KIND OF NEWER,
MORE NEWEL DISCOVERED MODE OF
MITOCHONDRIA FRAGMENTATION
THROUGH PERIPHERAL FISSION
OCCURRING IN THE OUTER PART OF
THE MITOCHONDRIA.
AND WHAT IT'S KNOWN IN
UNINFECTED CELLS FROM THE LAB,
IS THAT WHEN MITOCHONDRIA AND
THE PERILERAL FISSION, THIS IS
SUBJECTED TO MIRROR IMAGE TO
HAVAGEY SO IT'S TARGETED FOR
DEGRADATION AND IT HAS DECREASED
MEMBRANE POTENTIAL BUT I JUST
TOLD THAT YOU DURING INFECTION
THE OPPOSITE HAPPENS SO WILL
SHOWED IS THAT THIS FRAGMENT
MITOCHONDRIA ALL OF THIS BY
TRACKING IT, THEY BECOME
INCAPSULATED IN THIS STRUCTURE
IN MENC, AND THEY BECOME
PROTECTED SO HE SAW A DECREASE
IN MITOPHAGEY AND HOW THIS IS
DEPENDENT ON THE FORMATION OF
MENCs AND HE ALSO SHORED THIS
IS CONNECTED TO INCREASING
MEMBRANE POTENTIAL.
SO METHIS, Ys, PROTECTED AGAIN
MIRROR
MITOPHAGEY, OR
INTERRAMITOCHONDRIA CONTEXT
THROUGH THE ER AND YOU CAN SEE
THEM HERE AND THEN THEY FURTHER
COMMUNICATE BY STARTING TO SWAP
MEMBRANE POTENTIAL, YOU CAN SEE
THAT HERE.
THIS MITOCHONDRIA ER INCAP
SUEALATION CALLED MENC, IT WAS
FRAGMENT INDEED A POPULATION.
SO THEN WE WANT TO ASK WHETHER
THIS STRUCTURE COULD HAVE SOME
SORT OF BROADER ROLL VANCE, DID
WE JUST STUMBLE UPON THIS DURING
THE INFEC, IS IT MORE BROADLY
RELEVANT AND WHEN WE THINK ABOUT
ACMV, WE HAVE TO THINK ABOUT
DIFFERENT CELL TYPES, BECAUSE
ACMVs, KNOWN TO INFECT MANY
DEFINITE TYPES OF -- MANY
DIFFERENT CELLS AND OF COURSE
DEPENDING ON WHAT VIRUS TRAINS,
YOU'RE USING IN THE LAB, FOR
STUDYING THIS THEY EITHER HAVE
LOST OR STILL RETAIN THE ABILITY
OF INFECTING DIFFERENT TYPES OF
CELLS, SO WILL TACKLED THESE
QUESTIONS USING DIFFERENT HCMV
STRAIN USING EATER THE
FIBROBLAST OR EPITHELIAL CELLS
AND LOOK AT THEIR CHANGES AND
CHANGES IN MEMBRANE CONTEXT SIZE
AND USING A TARGETED MASS SPECT
TROMETRY ASSAY AS WELL AS DAY
TIME ACQUISITION ON A NEW
INSTRUMENT WE ACQUIRED AT ULTRA.
SO WHAT WE FOUND WAS INTERESTING
BECAUSE HE FOUND THAT EVEN FOR
THE CHANGE -- THE MEMBRANE
CONTEXT SITE, ALTERATION AND
MEMBRANE SITE THAT WERE
DIFFERENT IN EPITHELIAL CELLS OR
FIBROBLAST, THERE SEEM TO BE
SOME CONVERGENCE THERE, LET ME
TELL YOU WHAT I MEAN, SO IF WE
LOOK HERE AT THESE -- THESE ARE
DIFFERENT MEMBRANE CONTEXT SITES
THAT FACILITATE DIFFERENT
CONTEXT, BUT YOU CAN SEE HOW
THEIR ABUNDANCE STARTED AT A
DIFFERENT POINT IN UNAFFECTED
CELL IN EITHER FIBROBLAST OR
EPITHELIAL CELLS BUT THEN AS THE
INFECTION PROGRESSED IN THESE
DIFFERENT CELLS THEY STARTED TO
BECOME MORE SIMILAR TO EERCH
OTHER AND THE SAME HERE, RIGHT?
IT STARTED AS BEING HIGH IF NIEB
ROUGH ATOM BLAST AND LOW IN
EPITHELIAL CELLS BUT AS THE
INFECTION PROGRESSED, EVEN IF
THEY HAD DIFFERENT TRENDS, THEY
KIND OF ENDED UP WITH A SIMILAR
PROTEIN ABUNDANCE IN THESE
CELLS.
SO WHAT TELLS US IS EVEN IF THE
VIRUS IS IN FIBROBLAST AND
EPITHELIAL CELLS, THERE ARE SOME
CONVERGENT CHANGE THAT ARE
ALMOST PUSHING THE CELL TOWARD
SOME SORT OF UNIFIED CELL STATE
THAT IT'S AMENABLE AND SUPPORTS
WELL VIRAL INFECTION, AND THEN
THERE WERE ALSO SHARED CHANGES
THAT WERE THE SAME FOR ANY CELL
THAT WE LOOKED AT AND THESE ALSO
INCLUDED THE FORMATION OF
MENCs, SO AS I SAID EARLIER WE
ALREADY KNEW THAT THIS -- WHAT
MEMBRANE CONTEXT SITES ARE
NEEDED FOR FORMING THAT
STRUCTURE, THE MENC AND IT'S
PP51 ON THE MITOCHONDRIA SIDE
AND THE ON THE ER SIDE AND YOU
SEE HOW THE PROTEIN CHANGES
SIMILARLY IN ABUNDANCE IN
FIBROBLAST OR EPITHELIAL CELLS
AND IT SIMILARLY RECRUITED IN AN
ASYMMETRIC MANNER IN THIS MENC
STRUCTURE AND WE SAW THIS IN
FIBROBLAST, EPITHELIAL CELLS AS
WELL AS IN THE MACROPHAGE CELLS
SO NOW WE UNDERSTAND THAT OKAY,
IS THERE SOME BROAD RELEVANCE
THAT THE FORMATION OF MENCs IS
REALLY BROADLY IMPORTANT FOR THE
INFECTION IN DIFFERENT TYPES OF
CELLS, NOW THE OTHER QUESTION
WAS, WELL, HOW ABOUT MORE
BROADER CONTEXT AND OTHER
CONTEXT WHERE WE DO SEE
MITOCHONDRIA FRAGMENTATION BUT
IN CONJUNCTION WITH THIS HIGH
BIOGENICS AND IMMEDIATELY THAT
MADE US THINK OF CANCER BECAUSE
ACMV IS ALSO A VIRUS, AND WE
WILL SHOW THAT IT IS FORM
NOTHING MELANOMA CELLS AND
CRYSTAL IN MY LAB LOOKED AT HIGH
GRADE VARIANT CANCER AND FOUND
THE FORMATION OF MENCs AND
FURTHER MORE BY LOOKING AT
DIFFERENT MODEL SYSTEMS OF
TIEBAL OVARIAN CANCER AND
LOOKING AT LOOKING AT THOSE
ASSOCIATED WITH POOR PROGNOSIS,
SHE FOUND THESE ARE EXHIBITING
MENC IN CONJUNCTION WITH
DECREASE PHOSPHORYLATION,
DECREASE IMMUNE FACTORS AND
DECREASE IN ATV, SUGGESTS THAT
MENC'S BENEFICIAL FOR VIRAL
INFECTION AND PROMOTING OR OR
MAKING CHANGES IN THE CELLS THAT
ARE AMENABLE TO VIRAL INFECTION.
OKAY, SO WHAT I TOLD YOU SO FAR,
IS HOW OUR STUDIES OF ORGANELLE
REMODELING AND ORGANELLE,
ORGANELLE CONTEXT HAVE ALLOWED
US TO ANSWER THIS SORT OF
MARADOCKS IN THE FIELD OF ATMV
INFECTION, THE FACT THAT HOW
FRAGMENTED MITOCHONDRIA STILL
HAVE INCREASED OXIDATIVE
PHOSPHORYLATION AND IT'S THROUGH
THIS FORMATION OF MITOCHONDRIA
ER INCAP SUEALATION THAT
PROTECTS MITOCHONDRIA FROM
MIRROR IMAGE TO HAVAGEY AND
INCREASES RESPIRATION, BUT
ANOTHER OUTPUT OF THESE
METABOLIC CHANGES DURING
INFECTION AND IS PARTICULAR FOR
THIS FLEX IN GLYCOLYSIS IS AN
INCREASE IN LACTIC ACIDOSEISATE
PRODUCTION AND I WANT TO TELL
YOU HOW WE NOW UNDERSTAND HOW
LACTIC ACIDOSEISATE IS VERY
IMPORTANT IN THE INTRACELLULAR
AND INTRACELLULAR COMMUNICATION
AND PLACING THESE IN THE CONTEXT
OF VIRUS MICROVOIRMT.
SO WE KNOW THAT A NUMBER OF
VIRUSES ARE INDUCING METABOLIC
CHANGES THAT MIRROR WHAT HAPPENS
IN CANCER CELLS, IN PARTICULAR,
THEY INDUCE A WAR LIKE EFFECT
WHERE THERE'S A CHANGE THAT'S
AEROBIC GLYCOLYSIS WHICH IS
INCREASE IN LACTATE ON INFECTED
CELLS BUT OUTSIDE THE CONTEXT OF
INFECTION THERE HAVE BEEN
STUDIES IN CANCER RESEARCH THAT
HAS RECENTLY SHOWN THAT LACTATE
CAN ALSO INHIBIT IMMUNE
RESPONSES.
SO WE WONDER WHETHER ACMV
INFECTION, ALSO LEVERAGES THIS
INCREASE HEARING IN LACTATE FOR
IMMUNE SUPPRESSION AND IN ORDER
TO ADDRESS THIS, MA MATTE A
Ph.D. STUDENT IN MY LAB DID A
EXPERIMENT WHERE HE INFECTED
CELLS WITH A LOW MOI WHERE HE
LOOKED AT SPREAD OF INFECTION
AND IMMUNE SIGNALING AND HE
TREATED THESE CELLS WITH LACTATE
AND HE FOUND THAT IN THIS
CONTEXT OF MOI, LACTATE WAS
FUNCTIONING FOR THE BENEFIT OF
THE VIRUS WAS PROVIRAL FOR
SEQUENTIAL ROUNDS OF REP
WELLICATION.
WHAT DOES THIS MEAN?
WELL, FROM THE LAB OF MING AT
THE UNIVERSITY OF CHICAGO, WE
NOW IT CAN BE ADDED TO LYSINE
RESIDUES MODIFYING THESE
PROTEINS BY LACTATATION, SO
GIVEN THIS, WE KNOW ACMV INDUCES
LACTATE PRODUCTION, WE
ANTICIPATE THAT PROBABLY THERE
ARE EVENTS HAPPENING IN THE
CELL.
SO THEREFORE WE DID THE LACTATE
PURIFICATION, THIS IS ACTIVATED
LYSINE AT DIFFERENT TIME POINTSA
THE TIME OF INFECTION WITH ACMV,
AND THEN WE USE MASS SPEC
TROMETRY TO FIGURE OUT WHICH 1S
BECOME MORE INDUCED BY
MODULATION.
WE FOUND THERE ARE THOUSANDS OF
PROTEINS IN ALMOST EVERY
SUBCELLULAR COMPARTMENT THAT
BECOME LACTATED DURING
INFECTION, AND WHEN WE COMPARE
THIS WITH OTHER MODIFICATIONS WE
FOUND THE MOTIFS WERE DIFFERENT,
THE SITES WERE DIFFERENT AND
FOUND THAT LACTYLOME WAS
DEFINITE IN THESE REGIONS, SO
THEN WE THOUGHT IS THIS
SOMETHING WE NOTICE DURING
INFECTION WHEN WE COMPARE TO
OTHER FIBROBLAST DATA SETS AND
OTHER INFECTED STUDIES DURING
VIRAL INFECTION, OR HOW ABOUT
MORE BROADLY, SO WE DON'T LOAD
IT LACTYALATION, IN E-COLI AND
AGAIN WE FOUND THAT THE LACTATED
WERE ENRICH INDEED REGIONS OF
THE PROTEINS THAT WERE TREATED
TO BE INTRINSICALLY DISORDERS.
WE USE THOSE IN ALPHFOLD AND WE
CONFIRM THIS BY SEEING AN
INCREASE IN THE UNSTRUCTURED
REGIONS AND ALPHA HELIX
REPRESENTATION OF PEPTIDES IN
REGIONS OF THE PROTEINS THAT
CONTAIN THE LYSINE.
SO WHAT DOES THIS MEAN?
HOW IS LACTATATION OF A DISORDER
REGION HOW CAN IT IMPACT THEIR
FUNCTION?
SO WE LOOKEDDA THE FUNCTION OF
PROTEINS AS THE INFECTION
PROGRESSES DIFFERENT HOURS
DURING INFECTION.
SO CYTOKINE RESPONSE, CELLULAR
RESPONSE TO INTERFERON-GAMMA,
ACTIVATION OF INNATE IMMUNE
RESPONSES AND THEN WE ASKED IF
THIS IS JUST THE CASE FOR ACMV.
WE PERFORM SIMILAR LACTATION
STUDIES DURING HSV INFECTION AND
LOOKING AT VIRAL STRAINS THAT
ARE EITHER ABLE TO INHIBIT
IMMUNE RESPONSES OR UNABLE TO
INHIBIT IMMUNE RESPONSES AND
AGAIN, WE FOUND INCREASED
LACTYALATION, SO AGAIN THIS IS A
CONSERVED FEATURE IN INFECTION
EMPLOY SO AMONG THE IMMUNE
FACTORS WE FOUND TO BE MODIFIED
BY LACTYLATION, WHICH IS I5,
CYSTINE, THIS IS INTERFERON THAT
WE HAVE OTHER VS CHARACTERIZED
AS A VIRAL DNA SENSOR THAT CAN
FUNCTION IN THE NUCLEUS AND WE
FOUND THAT THIS PROTEIN BECOMES
DECORATED WITH LACTYLATION IN
ATV INFECTION AND HCMV INFECTION
AND SOME OF THESE ARE IN THE
REGIONS WE PREVIOUSLY
CHARACTERIZED.
SO LOOKING AT 1 OF THESE MUTATED
RESIDUES AND PERFORM MANY
TODAYS BUT I WANT TO FOCUS ON 1
OF THESE.
SO LOOKING AT 1 OF THESE SITES
WE FOUND THIS LACTATED DURING
BOTH OF THESE INFECTIONS WHEN HE
FOUNDS THIS TO A MIMIC HE
ACTUALLY NOTICED THAT THIS
PROTEIN STARTS TO HAVE INHIBITED
ABILITY TO INDUCE IMMUNE
SIGNALING SO THERE'S -- THERE
ARE DECREASES IN INTERFERON BETA
AND CXCL 10 AND IN AGREEMENT HE
SAW A INCREASE IN VIRUS TITER
WHEN THEY WERE EXPRESSING THIS
LACTIC ACIDOSEISILE MIMIC
PROTEIN.
DOWN STREAM ANOTHER IMMUNE
FACTOR DOWN STREAM OF DNA
SENSING RBN14, HERE IS
LACTYLATION THAT HE FOUND AND
AGAIN WHEN HE MUTATED THAT TO A
MIMIC HE FOUND THIS PROTEIN
STARTS TO HAVE INHIBITED ABILITY
TO INDUCE SIGNALING SO REDUCED
INTERFERON BETA AND CXCL-10
WOULD INCREASE VIRUS PRODUCTION
EMPLOY SO ADDRESS BROADER
IMPACTS IT IS HAPPENING, WHY
LACTYLATION OF HOST FACTORS
SEEMS TO SUPPRESS IMMUNE
SIGNALING WE HAD TO TURN BACK TO
PREVIOUS STUDIES IN WHICH WE TRY
TO PUT IN THIS BROADER CONTEXT
TO UNDERSTAND HOW IS IT WORKING
AND WHAT IS NEEDED FOR INDUCING
IMMUNE SIGNALING IN AND OVER THE
YORES WE HAVE DONE IN THIS A
NUMBER OF WAYS.
WE HAVE USED DIFFERENT METHODS
THAT ARE AVAILABLE TO US TO
STUDY LOCAL PROTEIN INTERACTIONS
OR GLOBAL PROTEIN INTERACTION,
AND ACTUALLY MORE RECENTLY, WE
MOVE MORE AND MORE TOWARDS
UNDERSTANDING OR CAPTURING
PROTEIN INTERACTION NETWORKS AT
THE SYSTEMS LEVEL, BECAUSE WE
STARTED TO APPRECIATE IF WE WANT
TO UNDERSTAND ORGANELLE
REMODELING AND HOW THINGS ARE
COORDINATED WE ACTUALLY NEED TO
CAPTURE AN INTERACTION NETWORK
MORE BROADLY.
AND FOR THIS, WE USE METHOD
CALLED THERMAL CO AGGREGATION
PROTILES, OF WHICH SOME CAN BE
CELLS OR TISSUES.
THE SAMPLE IS SUBJECTED TO
INCREASING TURNL AND FOR EACH
TEMPERATURE, YOU TAKE THE
REMAINING SOLUBLE FRACTION AND
ANALYZE IT BY MASS SPECTROMETRY
SO THE DAILY BASIS THEA WOULD
LIKE LIKE THIS, IT WOULD BE
ABNORMALITIES BROIGATION PROFILE
BECAUSE AS THE TEMPERATURE
INCREASES HAVE YOU LESS AND LESS
OF THE PROTEIN, IT BECOMES MORE
AGGREGATED SO HAVE YOU LESS IN
THE SOLUBLE FRACTION, BUT YOU
CAN LOOK AT THE SIMILARITIES OF
THIS CURVE TO START PREDICTING
INTERACTIONS BECAUSE WHEN
PROTEINS ARE TOGETHER AS A
COMPLEX, THEY DON'T BEHAVE AS
SEPARATE ENTITIES BUT RATHER
THEY BECOME THAT UNIT AND THEY
HAVE THE PROPERTIES OF THAT
COMPLEX UNIT.
SO THEREFORE THEY HAVE SIMILAR
ABNORMALITIES BROIGATION
PROTILES.
SO FEDERALLY WE USE IT TO STUDY
INTERN ACTIVITIES AND
PROJECTSIZATION RECEPTOR, WE
USED IT FOR HSV1 AND I WILL COME
BACK TO THAT AND WE MORE
RECENTLY USE IT TO STUDY AN
ASSOCIATED VIRUS BUT THE REASON
WHY THERE ARE LABS THAT
AREOOSING THIS TECHNIQUE IS
BECAUSE THESE AGGREGATION
PROFILES ARE A REALLY A MESS TO
STUDY, THEY ARE COMPLEX, IT'S
VERY HARD TO DECIPHER KIND OF
WHAT THIS MEANS IN TERMS OF
PROTEIN INTERACTIONS AND YOU
NEED REALLY GOOD COMPUTATIONAL
SKILLS AND POWER TO ANALYZE
THIS.
SO TAVIS REED, A Ph.D. IN MY
LAB, HE DEVELOPED A MACHINE
LEARNING COMPUTATIONAL PLATFORM
AND IT'S CALLED TAPIOCA, AND HE
DIDN'T UNDERSTAND WHAT TPCA
MEANS SO HE CALL TODAY TAPIOCA,
AND I SAID OKAY, WE WILL CALL IT
TAPIOCA, SO TO USE LOGISTIC
MODELS TO INTEGRATE SUBSETS OF
DYNAMIC DATA FROM TPCA WITH
DIFFERENT STATIC DATA FROM
TISSUE SPECIFIC FUNCTIONAL
NETWORKS AND DOMAINS OF THE
PROTEINS AND PREDICTED PHYSICAL
PROPERTIES OF THE PROTEINS AND
HE SHOWED HOW USING THIS, YOU
CAN DECONVOLUNTEERSUTE THIS
COMPLEX DATA AND IT DOESN'T
MATTER HOW FULLY ABOUT HAIFED
THE PROTEIN IS AND YOU CAN USE
THIS TO PREDICT DENOVAE
INTERACTIONS AND COMING BACK,
WHY THAT MATTERED TO OUR IL516
STORY AND LACTYALATION STORY, IS
THAT WE USE THAT TO UNDERSTAND
THE UNDERACTION AND IT ALLOWED
US TO CHARACTERIZE IT.
SO TO SUMMARIZE A BIT, WE AND
OTHER VS SHOWN THAT THIS CAN
SENSE NUCLEAR VIRAL DNA AND HE
DOES THAT BY LOCALIZING THE
PERIPHERY HERE, AND INSERTS THE
VIRAL DNA HERE, AS IT LOCALIZES
HERE.
IN MORE RECENT YEARS WE SHOWED
HOW I516 IN THE PERIPHERY
RECRUIT THE DNA DAMAGE WHICH IN
TERMS PHOSPHORYLATES IFI16 IN
THIS DISORDER REGION THAT WE
JUST TALKED ABOUT.
AND THAT PHOSPHORYLATION WAS
SHOWN TO BE NECESSARY FOR THE
ABILITY TO INDUCE IMMUNE
SIGNALING SO IT ADDS TO THIS
GROWING UNDERSTANDS OF ACROSS
TALK BETWEEN DNA SENSING AND DNA
DAMAGE RESPONSE IN IMMUNE
SIGNALING.
SO WE ALREADY KNEW THAT THIS
INTRINSICALLY DISORDER REGION IS
SO IMPORTANT FOR THE ABILITY OF
THIS TO INDUCE IMMUNE SIGNALING
AND THAT IT'S DECORATED WITH
THIS PHOSPHORYLATION SITE AS
WELL AS ADDITIONAL
PHOSPHORYLATION SITE AND THAT
WAY IN MY LAB AND MORE RECENTLY
HAS SHOWN THAT THOSE NUMEROUS
PHOSPHORYLATIONS IN THE
INTRINSICALLY DISORDERED REGION
CAN EXPLAIN SOMETHING WE NOTICE
FOR MANY YEARS FOR THIS PROTEIN,
THAT ONCE IT BINDS TO VIRAL DNA,
IT HAS THESE REALLY STRIKING 3
DYNAMIC BEHAVIOR THAT STARTS TO
FORM THIS STRUCTURE AND WE
COULDN'T UNDERSTAND WHAT THAT
MEANS.
AND DAWEI SHOWED THAT THIS
HAPPENS ON THE LESION, AND IT
UNDERGOES SEPARATION AND THIS
DYNAMIC BEHAVIOR AND HOW THIS IS
IMPORTANT FOR INDUCING SIGNALING
SO WE KNOW IN THAT REGION, THAT
DISORDER REGION IS IMPORTANT AND
REGULATED BY POST TRANSLATIONAL
MODIFICATIONS AND NOW SUDDENLY
WE KNOW THAT IT'S ALSO
LACTYLATATED AND HOW IT CAN
SUPPRESS THE CYSTINE IMMUNE
SIGNALING.
SO THEREFORE WE WENT BACK TO OUR
TPCA DATA SET TO FIND OUT WHAT
INTERACTIONS ARE AFFECTED BY THE
LACTYLATION SITE AND IN LOOKINGA
THE THIS, IT ACTUALLY WHEN WE
USE THE LACTY-MIMIC, IFI16 HAS
IMPEDED ABILITY TO RECRUIT THE
DNA PK AT THE NUCLEAR PERIPHERY
AND AS I TOLD YOU THE DNA IS
NEEDED FOR POSFORALATING IK516,
SO NOW WE UNDERSTAND THAT
LACTATE IS VIRAL FOR SEQUENCE
ROUNDS OF REPLICATION AND IN
PART, THIS IS THROUGH THE HOST
IMMUNE FACTOR EXPTION
SUPPRESSION EVER CYTOKINE
EXPRESSION AND WE STARTED TO
LOOK AT WHAT ENZYMES ARE
REGULATING THIS AND THAT'S KIND
OF ANOTHER STORY.
BUT I WANT TO SPEND A BIT OF
TIME, THE REMAINING PART OF TIME
TO START TO PUT THESE FINDINGS
IN THE BROADER PICTURE BECAUSE
WHAT I TOLD YOU ABOUT LACTATE IT
BROUGHT A NEW PERSPECTIVE TO OUR
LONG-TERM INTEREST IN
UNDERSTANDING HOW ALL THESE
CHANGES THAT I'M TELLING YOU
ABOUT IN THE INFECTED CELL, HOW
ARE THEY CONVAIPED TO THE
ENVIRONMENT OF THE CELL, TO THE
CELLS THAT ARE IN THE IMMEDIATE
PROXIMITY OR THOSE THAT ARE
FURTHER AWAY, HOW IS THIS
INFECTED CELL CHANGING ITS
ENVIRONMENT.
AND THE LACTATE OF COURSE PLAYS
A ROLE IN THIS.
SO A FEW YEARS AGO WE TRY TO
ADDRESS THIS TO DESIGN A METHOD
FOR SEPARATING THE CELLS BASED
ON HOW CLOSE OR FAR THEY ARE
FROM AN INFECTION SITE TO REALLY
TART TO TACKLE THIS QUESTION OF
THE VIRUS MICROENVIRONMENT, TO
SEPARATE THE CELLS AND START TO
CHARACTERIZE THEM AND IN ORDER
TO DO THIS, WE ADAPTED A METHOD
THAT WAS PREVIOUSLY USED FOR
STUDYING THE TUMOR
MICROENVIRONMENT, SO LAWEDDER
DALE WE DID WAS TO GENERATE
FIBROBLASTS, THAT EXPRESS AN
M-CHERRY PROTEIN, SECRETARY
PEPTIDE AND MAKING THESE RED
CELLS, WHICH WE THEN INFECTED
WITH 1 OF OUR GREEN FLUORESCENT
VIRUS, CREATING THESE DULY
FORESTIMATE THADENT CELLS, RED
AND GREEN.
AND THEN WE CAREFULLY OPTIMIZE
THE COCULTURES OF WILD-TYPE
CELLS IN AN ORDER TO ALLOW FOR
THE SPREAD OF THIS MCHERRY
SIGNAL PEPTIDE, PRIOR TO THE
SPREAD OF AN INFECTION, SO IN
OTHER WORDS, WE WILL HAVE THE
INFECTED CELLS THAT ARE GREEN
AND RED, THE NEIGHBORS CELLS
THAT ARE RED AND THOSE THAT ARE
FURTHER AWAY, THAT ARE NOT
LABELED.
AND THEN WE CAN USE FLUORESCENCE
ACTIVATED CELL SORTING TO SORT
THE CELL POPULATION AND THEN USE
PROTEOMICS THAT USE MASS
SPECTROMETRY TO CHARACTERIZE THE
COMPOSITION BUT ALSO DO
FUNCTIONAL STUDIES ON THE
SEPARATED CELL POPULATIONS.
SO WHAT WE DISCOVERED WAS THAT A
FEW REALLY INTERESTING THINGS,
SO IN THE INFECTED CELL AS WE
EXPECTED ACMV HAS MECHANISMS FOR
SUPPRESSING IMMUNE SIGNALING SO
WE FOUND THAT IMMUNE FACTORS ARE
DECREASED HOWEVER, WE FOUND THAT
THE NEIGHBORING CELLS ALSO HAD
DAMPENED IMMUNE SIGNALING AND
THAT THE DISTAL CELLS WERE THE
1S THAT ACTUALLY HAD ENHANCED
IMMUNE FACTORS.
THE MEMBRANE CONTEXT SITES THAT
I TOLD YOU ABOUT EARLIER, THAT
REPRESENT ALL THOSE ORGANELLE,
ORGANELLE CONTACTS, WE FOUND
THEY WERE REMODELED IN INFECTED
CELLS BUT WE ALSO FOUND THAT THE
SPECIFIC SUBSET OF THESE
MEMBRANE CONTEXT SITES WERE ALSO
CHANGED IN NEIGHBORING CELLS CAN
AND THEN THEY WERE NORMAL AND
DISTAL.
AND FINALLY PROBABLY THE MOST
PUZZLING 1 IS YES, WE KNOW THAT
VIRUSES TARGET THE CELL CYCLE
AND THAT THEY ARREST ACMV FOR
EXAMPLE, ARREST CELLS AT G1 AS
INTERPHASE IN ORDER TO INSURE
ITS NUCLEAR DNA REPLICATION, BUT
WE FOUND THAT THE NEIGHBORING
CELLS GET ARRESTED IN MITOSIS IN
M-PHASE.
SO NOW WITH THE STORY I TOLD YOU
BROUGHT A NEW PERSPECTIVE FROM
THIS, NOW WE UNDERSTAND THAT IT
CAN BE ADDED TO IMMUNE SIGNALING
TO IMMUNE FACTORS AND THAT CAN
ALSO THE FACT THAT IT'S SECRETED
THEN PRODUCING SUCH LARGE AMOUNT
CANS DAMPEN THE IMMUNE RESPONSE
SO PROBABLY IT CONTRIBUTED TO
THIS EFFECT THAT WE SEE HERE.
BUT WE ALSO WONDERED, WELL, WHAT
ABOUT ALL THESE CHANGES IN
NEIGHBORING CELLS THAT SEEM TO
BE SOMEHOW PRIMED WITH ALL THESE
CHANGES.
WHAT IS THE RELEVANCE OF THESE
CHANGES TO CO INFECTIONS SORE TO
DISEASE PROGRESSION AND THE
REASON WE ARE INTERESTED IN CO
INFECTION AS I SAID, ACMV INFECT
SO MANY DIFFERENT CELL TYPES AND
SO MANY TISSUES AND IS KNOWN
THAT THOSE CAN BE INFECT WIDE
MANY OTHER VIRUSES AND THERE'S A
LOT OF INTEREST OF COURSE, IN
THE LINK BETWEEN CO INFECTIONS
AND MORBIDITY AND DECREASED
IMMUNITY, CANCER DEVELOPMENT AND
SO ON.
SO WHAT WE DID WAS TO GENERATE
AGAIN, OUR VIRUS,
MICROENVIRONMENT AND THEN TEST
THE SUSCEPTIBILITY TO INFECTION.
SO THIS IS A VIRUS
MICROENVIRONMENT FROM ACMV, AND
WHAT WE FOUND IS THAT CELLS
NEIGHBORING THE SITE OF ACMV
INFECTION ARE MOSTLY SUSCEPTIBLE
TO INFECTION NOT ONLY WITH ACMV
BUT ALSO WITH HSV1, AND ALSO
WITH INFLUENZA A.
SO ANOTHER NUCLEAR REPLICATING
VIRUS BUT AN RNA VIRUS.
WE INITIALLY DID THIS LOOKING AT
FIBROBLASTS SURROUNDED BY
FIBROBLASTS SO AT LEAST THE
CELLS BACKGROUND IS KEPT
CONSISTENT BUT THEN WE STARTED
TO ALSO LOOK AT CELLS SURROUNDED
DIFFERENCE INTERACTIONS BETWEEN
FIEBLOBLAST AND EPITHELIAL CELLS
AND WE FIND AGAIN THAT
EPITHELIAL CELLS SURROUNDED ARE
MORE SUSCEPTIBLE TO INFECTION
WITH VIRUSES AS I'M SHOWING
HERE.
SO CLEARLY VIRAL INFECTION
RESHAPING THE MICROENVIRONMENT
TO FACILITATE PROGRESSION OF
INFECTION AND WE START TO HAVE
EVIDENCE THAT SOME OF THESE
CHANGES MAY BE LINKED TO
PROPERTIES BECAUSE WE HAVE
SCIENCE THAT THERE MAY BE
GENOMIC INSTABILITY IN THESE
CELLS AND ALSO THE CELL CYCLE
DISTURBANCE.
SO THEREFORE GOING FORWARD, WE
STARTED NOW TO ADOPT THIS TO
LOOK AT CELL TO CELL
COMMUNICATION AS IT OCCURRED IN
THE TISSUE, SO LOOKING AT DIRECT
CELL TO CELL CONTACTS SO CRYSTAL
IN MY LAB, LOOKING FOR RIKE
ROUGH ATOM ENVIRONMENT, AND
SHOWING 1 EXAMPLE HERE IN
PATIENT DERIVED COLORECTAL
CANCER ORGANOID NCH YOU CAN SEE
HOW SHE CAN MONITOR DIFFERENT
STAGES OF AN INFECTION IN THIS
ORGANOID AS AS IT PROGRESSES AND
THEN SHE WENT AHEAD TO PROGRESS
THESE CELLS TO CHARKTIZE THEM.
SO SINCE I WANT TO LEAVE A LOT
OF TIME FOR QUESTIONS, I AM
GOING TO FINISH TO SAY THAT I
HOPE I SHOWED YOU HOW WE CAN
INTEGRATE MOTE I DON'T MEANIC
AND VIROLOGY STUDIES AND
MICROSCOPY STUDIES TO START TO
LOOK AT CELLULAR COMMUNICATION
DURING INFECTION AND THAT WE
LOOKED AT CELLULAR COMMUNICATION
EITHER AT THE INTRACELLULAR, OR
INTRACELLULAR LEVEL, WE LOOKEDDA
THE ORGANELLE, ORGANELLE
CONTACTS THAT ARE FACILITATED
AND/OR FACILITATING ORGANELLE
REMODELING AND HOW THIS LED TO
THE DISCOVERY OF THIS
MITOCHONDRIA HERE INCAPUE
SUEALATIONS THAT WE CALLED --
INCAPUIZATIONS CALLED MENCs,
AND THEY INCREASE AND MAINTAIN
PHOSPHORYLATION, I SHOWED YOU
HOW THE CHANGES THAT HAPPEN IN
METABOLISM, ARE INDUCING ALSO
INCREASED LACTATE PRODUCTION AND
ALSO WHY THIS IS IMPORTANT AT
THIS INTERFACE BETWEEN
DISRUPTION OF METABOLISM AND
IMMUNE SIGNALING BECAUSE THIS
INCREASED LACTATE PRODUCTION
DRIVES LACKAATION OF HOST IMMUNE
FACTORS OF THESE REGIONS AND
THIS INHIBITS THEIR ABILITY TO
INDUCE CYTOKINE PRODUCTION, AND
HOW WE LEARN WHAT SPECIFIC
PROTEIN INTERACTIONS ARE
DISRUPTED USING THESE THERMAL
PROFILING AND INTERACTION
NETWORKS, FINALLY, I SHOWED YOU
HOW WE STARTED TO LOOK AT HOW
THIS INFORMATION IS CONVEYED TO
THE ENVIRONMENT AND WHAT'S THE
IMPACT OF AN INFECTED CELL ON
ITS NEIGHBORHOOD, AND HOW WE
FOUND THAT NEIGHBORING CELLS, SO
CELLS NEIGHBORING IN THE
INFECTED SITE WITH CELLS INFEBT
WIDE ACMV, BECOME MORE
SUSCEPTIBLE TO INFECTION WITH
HSV1 AND SCMV AND EVEN INFLUENZA
A AND WE ARE ARE PERFOR THE
PURPOSING QUITE A FEW STUDIES --
PERFORMING QUITE A FEW STUDIES
WITH THESE MECHANISMS NOW.
SO WITH THAT I WANT TO THANK MY
LAB FOR ALL THEIR TREMENDOUS
WORK, THE FUNDING INCLUDING
FUNDING FROM THE NIH, FROM NIAID
AND NIGMS, I RECENTLY BECAME AS
YOU HEARD, LAST YEAR, I BECAME
THE EDITOR AND CHIEF OF MCP AND
INVITE YOU TO A SUMMER
CONFERENCE THAT WILL BE IN
BOSTON AND I WANT TO FINISH BY
SAYING THAT ALSO VERY RECENTLY
WE LOST ANOTHER PHENOMENAL
VIROLOGIST DR. THOMAS KRISTIE,
WHO I HAD THE OPPORTUNITY TO
WORK WITH AND COLLABORATE AND
CALL MY FREEND AND HE WILL BE
ABSOLUTELY DEARLY MISS AND I
JUST WANT TO SEND MY THOUGHTS
AND MY APPRECIATION FOR
EVERYTHING HE HAS DONE FOR THIS
COMMUNITY.
SO THANK YOU SO MUCH.
[ APPLAUSE ]
>> WELL WE HAVE MICs, WE HAVE
PLENTY OF QUESTIONS ONLINE.
CAN YOU TRIBUTE THE DIFFERENT
VIRUS EFFECTS TO THE LEVELS OF
THE TYPES OF INTERFEREONSS
INDUCED?
>> DIFFERENT VIRUS EFFECTS TO
TYPES OF INTERFERONS, JUST TO
CLARIFY ABOUT WHICH 1 IS THE
QUESTION -- WHAT IT REFERS TO.
>> IT CAME IN RELATIVELY EARLY
IN YOUR TALK AND IT'S FROM AN
IMMUNOLOGIST.
>> YES, SO THE TYPE OF VIRUSES,
THE TYPE OF VIRUSES, SAY AGAIN.
NCAN YOU ATTRIBUTE THE DIFFERENT
VIRUS EFFECTS TO THE LEVELS OF
TYPES OF INTERFERONS INDUCED?
>> OKAY, SO, IF IT'S IN THE
FIRST PART IT'S PROBABLY ABOUT
ORGANELLE REMODELING AND HOW
THOSE DIFFERENT CHANGES IN
ORGANELLE REMODELING CAN ALSO
IMPACT IMMUNE SIGNALING.
SOIME GOING TO SPECULATE AND
ANSWER THIS IN MANY DIFFERENT
WAYS BECAUSE I CAN ANSWER FROM
DIFFERENT PARTS.
>> I KNOW THIS PERSON I KNOW AND
MAYBE HE WILL WRITE BACK TO
CLARIFY.
>> YES, GREAT.
SO I CAN ALSO MAYBE JUST SAY
SOMETHING ABOUT IMMUNE SIGNALING
AND ORGANELLE REMODELING BECAUSE
THAT'S SOMETHING WE WERE
THINKING EACH LET'S SAY FROM THE
FIRST PART ON MITOCHONDRIA
REMODELING, YOU KNOW, WE WERE
LOOKING AT THESE MITOCHONDRIA ER
CONTACTS AND AS YOU KNOW, A
RESIDENT OF ER IS IMPORTANT
PROTEIN THAT IS IMPORTANT FOR
IMMUNE SIGNALING, SO, WE WERE
THINKING OKAY, THESE METHIS, C
FORMATIONS ARE ANTIVIRAL BUT IF
WE -- IF WE INDUCE THIS CONTACT
BETWEEN MITOCHONDRIA AND THE ER
EARLY ON, IT'S NOT ANTIVIRAL,
IT'S ACTUALLY PROVIRAL BECAUSE
IT ACTIVATES STING.
SO IF MITOCHONDRIA IS NOT YET
FRAGMENTED IT'S FILLA
METROPOLITAN OS ININATE AND YOU
ARE YOU'RE FEDDERRING ARTIFICIAL
TO THE ER, IT ACTIVATES STING.
SO THAT'S YOU KNOW -- SO THAT
QUESTION IS ABOUT WHETHER
ORGANELLE REMODELING AND
MITOCHONDRIA FRAGMENTATION IS
ALSO TO SUPPRESS IMMUNE
SIGNALING.
- IT IS A POSSIBILITY.
IT'S ALSO POSSIBLE THERE'S JUST
SOMETHING TO DO WITH MOTILITY
AND SO ON.
BUT MY UNDERSTAND SUGGEST THAT
THE CLARIFICATION WILL COME
PRETTY SOON.
YES.
>> FANTASTIC TALK.
I'VE ALWAYS LOVED -- YOUR
GROWING STORY ON LOOKING AT
INFECTED VERSUS NEIGHBORING
VERSE US DISTANT CELLS AND
REALLY FASCINATING SO WITH THE
NEIGHBORING CELLS YOU ALSO SEE
THE MENC CHANGES.
SO IS THAT, YOU TOUCH OFFICE OF
DIVERSITY THIS A -- YOU TOUCHED
ON THAT A LITTLE BIT MAYBE BUT
IS THAT FROM VIRUS UPTAKE,
METABOLISM, DAMPENING IMMUNE
RESPONSE, HOW DIFFERENT IS THAT
UNINFECTED NEIGHBORING CELL FROM
AN INFEBTED CELL?
>> SO IT'S QUITE DIFFERENT AND
THERE SEEM TO BE ONLY A SUBSET
OF THOSE MEMBRANE CONTEXT SIDES
THAT ARE CHANGED AND SOME OF
THEM SEEM TO BE WITH WITH
REGULATING TRANSFER SO WE SEE
ONLY SPECIFIC POOLS OF MEMBRANE
CONTEXT SIZE THAT ARE CHANGED IN
UNINFECTED CELLS THAT ARE
NEIGHBORING IN INFECTED CELLS
THEY LOOK QUITE DIFFERENT SO
IT'S SOMETHING TO DO EXACTLY AS
YOU SAY WITH PRIMING THOSE
NEIGHBORING CELLS FOR INFECTION,
SO THAT'S WHY WE HAVE OTHER
PROJECTS IN THE LAB WHERE WE ARE
LOOKING NOW AT HOW THESE ARE --
CONTRIBUTING TO CHANGES IN
CELLULAR STATE.
BECAUSE WE KNOW OF COURSE,
VIRUSES CAN INDUCE MESENCHYMAL,
TRANSLATION AND SOMETHING WE
WERE STUDY NOTHING THE CONTEXT
OF ACMV INFECTION, BUT OR THE
OPPOSITE EMT, BUT HOW DO THEY
CHANGE ALSO THE STATE OF THESE
NEIGHBORING CELLS AND FURTHER
MORE, AND THAT AS YOU CAN
IMAGINE CAN BE QUITE HAS THIS
IMPLICATIONS AND PATHOLOGIES.
YEAH.
>> HI, ILEANA, SO WITH REGARDS
TO THE MENC PART OF THE STORY.
TWO QUESTIONS, 1, WHEN THEY
FIZZ, THE 25% FISSION ON THE
END, DO YOU SEE LOSS OF
MITOCHONDRIA POTENTIAL BEFORE
THE FISSION OR IS THE MENC JUST
HELPING TO MAINTAIN IT INSTEAD
OF STORE THE MITOCHONDRIA
POTENTIAL AND ALSO WHAT'S THE
BENEFIT FOR THE VIRUS IN MAKING
SMALL BUT HIGHLY BIOENERGETIC
VERSUS MITOCHONDRIA, VERSUS
FUSED MITOCHONDRIA.
>> BEAUTIFUL QUESTION.
VERY BEAUTIFUL QUESTION.
YES, WE DO SEE A SLIGHT DECREASE
IN THIS AND THEN WE SEE THE
INCREASE THE RESCUE, RIGHT?
IN TERMS OF -- AND IT'S
INTERESTING BECAUSE WE'VE BEEN
MONITORING THIS AND TRYING TO
LOOK AT TEMPORALITY AND EVEN FOR
THE SWAPPING OF MEMBRANE
PROTENTIAL SO IT'S INTERESTING
HOW IT GOES AWAY AND COMES BACK.
BUT IN TERMS OF THE RATIONAL FOR
THIS, I THINK IT'S -- IT'S --
THERE ARE MULTIPLE
POSSIBILITIES, SO OF COURSE,
SOME OF THIS IS KNOWN TO BE
CONNECTED TO REGULATION OF
APOPTOSIS, BUT OTHER CAN BE ALSO
WITH MOTILITY AND WHERE THIS
MITOCHONDRIA ARE BEING LOCALIZED
BECAUSE WE START TO SEA THEM,
HOW LATER IN INFECTION, THEY GO
AROUND THE ASSEMBLY COMPLEX
WHERE YOU GO ALIFE ON EARTH THE
OF THAT REARRANGEMENT OF GOLGI
AND THE ENDOSOMES, ALL THAT
HAPPENS, ORGANIZATION AND
PRODUCTION OF NEW INFECTIOUS
VIRAL PARTING, SO MAYBE ENERGIES
BROUGHT THERE AT THE SITE WHERE
IT'S NEEDED, IT'S VERY POSSIBLE
THAT'S 1 LIKELY SCENARIO TO BE
DEMONSTRATED, YEAH.
>> I HAD ANOTHER MENC QUESTION,
SO YOU TALK ABOUT THE FORMATION
OF THESE STRUCTURES AND HMV
INFECTION AND IN CANCER, ARE
THERE NONDISEASE STATES IN WHICH
THESE STRUCTURES ARE FORMED
DURING DEVELOPMENT OR SOME OTHER
SCENARIO?
>> SO YEAH, REALLY GOOD POINT,
CONFECTED TO THE EARLIER
QUESTION AS WELL, I'M GLAD YOU
ASKED IT, THE REASON WE ARE
LOOKING AT THOSE IS BECAUSE WE
SAW MENCs BEING FORMED IN THE
SMALL LEVEL IN UNINFECTED CELLS
SO WE KNEW THEY COULD FORMED BUT
INFECTION PROMOTE THIS IS IS
PUSHES THIS MITOCHONDRIA INTO
THIS PARTICULAR STATE BUT THEY
CAN EXIST.
SO, THAT'S WHYY WOO THOUGHT,
OKAY, THEY EXIST ALSO IN
UNINFECTED CELLS AND THEN NOW WE
KNOW, AND THAT THEY ARE EVEN
CONNECTED TO POOR PROGNOSIS AND
I WOULD SAY THAT YES, IN TERMS
OF YOUR LAST QUESTION, WE
STARTED TO LOOK AT SOME OF THE
NEURAL DEGENERATIVE DISEASES
LIKE ALZHEIMERS AND HUNTING
TON'S AND SO ON AND PERHAPS IN
ALZHEIMER, SO IT'S POSSIBLE,
IT'S POSSIBLE THIS IS MAY BE A
MECHANISM TO PROTECT THE
MITOCHONDRIA THAT NEEDS TO
PRODUCE A LOT OFOXIDATIVE
PHOSPHORYLATION, YOU KNOW
PRODUCE ENERGY, IT'S POSSIBLE,
IT JUST REMAINS TO BE SEEN.
WE STARTED TO LOOK INTO THAT A
LITTLE BIT BUT IT'S JUST VERY
RECENTLY.
YEAH.
ONE THING I WILL MENTION IS
THAT, AND KIND OF CONNECTED TO
ACTUALLY -- I WILL ANSWER YOUR
QUESTION FIRST.
>> WELL, I HAVE 3.
>> PERFECT.
THEN YOU MAY EXACTLY WHAT I WAS
GOING TO SAY.
>> TWO TECHNIQUESTIONS TO START
WITH.
FOR THE FORMATION OF MENCs,
HAVE YOU IDENTIFIED ANY RNA
MARKERS WHERE PEOPLE HAVE LOOK
AT THE RNA LEVEL TO SEE IF THAT
SUGGESTS INCREASED FORMATION OF
MENCs?
>> NO, SO WE HAVE NOT.
NO, BUT THE DETECTION OF THE
MCSs THEMSELVES ISSUES THE 1S
WE FOUND TO BE LOCALIZED THERE
IS REALLY, THEY PROVIDE A GREAT
MARKER BECAUSE THIS IS 1 OF
THOSE FEW EXAMPLES WHERE A
CHANGE IN THE ABUNDANCE OF A
PROTEIN REALLY SIGNIFIES A
CHANGE IN THE EXTENT OF CONTACT
BETWEEN 2 ORGANELLES, SO BECAUSE
USUALLY, YOU KNOW YOU FIND
CHANGES IN ABUNDANCE AND WHAT
DOES THAT MEAN, IT CAN MEAN A
LOT OF DIFFERENT THINGS, BUT
THEY PROBABLY ARE ALSO OTHER
MARKERS THAT ARE NOT AT THE RNA
LEVEL BUT RATHER THE PROTEIN
LEVEL, I WOULD NOT BE SURPRISED
IF SOME LET'S SAY POST
TRANSLATION MODIFICATION ARE AND
ARE IMPORTANT FOR TARGETING,
IT'S KNOWN THAT HAPPENS FOR
CERTAIN MCSs, THAT THEY NEED
TO BE MODIFIED IN CERTAIN WAYS
IN ORDER TO BE RECRUIT INDEED A
CERTAIN CONTACT, FOR PROTEINS
THAT HAVE MULTIPLE
LOCALIZATIONS.
YEAH.
>> OKAY AND ARE THERE ANTIBODIES
THAT ARE NOW SPECIFIC TO
LACTYLIEE, AUDIENCE TION SITES
IN PROTEINS, ARE THESE REAGENTS
THAT PEOPLE CAN THINK ABOUT
USING?
>> THERE ARE ANTIBODIES THAT ARE
SPECIFIC TO LACTYLATED LYSINE,
NOT NECESSARILY TO A SITE IN A
PROTEIN AND THE INITIAL STUDIES
ON LACTYALATION, INITIAL STUDIES
HAVE BEEN DONE ON HISTONE
MODIIVESS AND HOW, YOU KNOW
NUCLEAR LACTYALATION IS
IMPORTANT AND WHY THIS IS
IMPORTANT JUST IN TERMS OF
POSSIBLE EVEN CROSS TALK WITH
OTHER PTMs ON HISTONES AND
GENE EXPRESSION.
SO, THIS IS KIND OF HOW THAT
FIELD OR POST TRANSLATION
LACTYALATION CAN BE REGULATED
AND WE HAVE STARTED TO LOOK AT
WHO ENZYMES IN THE CONTEXT OF
INFECTION ARE REGULATING THIS
MODIFICATION, IT'S FASCINATING
WE FOUND THAT AN ENZYME THAT IS
INVOLVED IN PLACING ALA9 AND SO
TRNA, SO ALAN INE, IS THE
ONESELF THAT BRINGS THE LACTATE
ON PROTEINS SO YOU CAN PUSH THIS
ENZYME IF YOU FEED CELLS AA
NINE, AND IF YOU FEED THEM
LACTATE SO IT'S CALLED AARS 1
THAT IT FUNCTIONS AND WORKS IN
THE NUCLEUS AS WELL AND ARS2
THAT FUNCTIONS IN THE
MITOCHONDRIA FOR SOME OF THE
LACTYALATIONS THAT WE'VE SEEN.
>> I SEE, 1 MORE QUICK QUESTION.
IN THE LAST PART OF YOUR TALK,
YOU MENTIONED THAT THE
NEIGHBORING CELLS HAD A DAMPEN
IMMUNE RESPONSE ESTATE BUT THE
CYSTAL 1S HAD AN INHANSED STATE,
DO YOU HAVE IDEAS OF THE
SIGNALING THAT'S HAPPENING
BETWEEN THE NEIGHBORING CELLS
AND THE DISTAL CELLS?
>> NO, BUT WE STARTED TO LOOK AT
THIS AND ALSO FROM THE
PERSPECTIVE OF HETEROGENEITY OF
RESPONSES SO WITHIN EACH OF
THESE POPULATIONS AS CAN YOU
IMAGINE, LET'S SAY THE
NEIGHBORING, THE DISTAL, THERE'S
STILL SUCH HETEROGENEITY OF
VIRAL INFECTION AND DIFFERENT
SUSCEPTIBLE TO INFECTIONS, SO WE
STARTED NOW TO SEPARATE THESE
AND A BIT MORE THAN A YEAR AGO,
WE INSTALLED AN INSTRUMENT IN
THE LAB THAT CAN DO SINGLE CELL
PROTEOMICS SO NOW WE'RE
SEPARATING ALL THESE CELLS AND
ARE LOOKING AT THE SINGLE CELL
LEVEL KIND OF WHAT COMPOSITION
REPRESENTS A CERTAIN RESPONSE OR
ABILITY AND WE ARE GETTING TO
THAT QUESTION OF INTERACTIONS
BETWEEN THE NEIGHBORING AND THE
DISTAL CELLS.
>> GREAT, THANKS.
>> BACK TO THE LACTYALATION,
THAT IT REVENTS THE RECRUITMENT
OF KINASES?
DO YOU KNOW THE MECHANISM FOR
THAT?
IS ITSTERRIC HINDRANCE OR
SOMETHING MORE SUBTLE GOING ON?
>> THAT DISORDER REGION WE FIND
TO BE MODIFIED WE FIND THAT MANY
TIMES WHAT HAPPENS TO THAT AREA
OF THE REGION HAS A LOT TO DO
WITH CHARGE, CHARGE-CHARGE
INTERACTIONS AND MAYBE THAT'S
WHAT IT'S ABOUT, AND HOW IT'S
SUDDENLY FOLD IN AND CREATES A
DIFFERENT FOLD ENVIRONMENT.
SO WE DO IN THE KNOW MORE, ALL
WE KNOW FOR THE MOMENT IS THAT
WHEN THE LACTIC ACIDOSEISERATION
IS PRESENT ON THE DISORDER
REGION SUDDENLY IT CAN NO LONGER
BE RECRUITED TO THE PERIPHERY OF
SITES OF THE GENOME POSITION,
THAT'S WHEREY WOO ARE WITH THAT
FOR THE MOMENT.
ANY EFFECT ON PROTEIN STABILITY.
NO.
WE DID NOT SEE AN EFFECT ON
PROTEIN STABILITY BUT I WANT TO
ALSO MENTION SOMETHING ELSE ON
LACTIC ACIDOSEISALATION BECAUSE
JUST FOR THE FLOW OF THE TALK
AND INTEREST OF TIME, I WANTED
TO GIVE YOU ONLY THE STORY ABOUT
THE IMMUNE SIGNALING AND THE
LACTATION ON'MUNE FACTORS, YOU
CAN'T SAY THAT SOMETHING IS
ANTIVIRAL PROVIRAL BECAUSE YOU
ARE GOING TO HAVE THOUSANDS OF
PROTEINS AND THEY WILL HAVE SO
MANY FUNCTIONS SO WHAT YOU ARE
LOOKING AT IS REALLY AN
AVERAGING OF THAT.
BUT WE ALSO FOUND VIRAL PROTEINS
THAT BECOME LACTATED AND SOME OF
THESE VIRAL PROTEINS WHEN WE DID
THE SAME EXPERIMENT, I SHOWED
YOU EARLIER WITH LACTATE
TREATMENT AND LOW M OY, AND WE
FOUND THAT IT WAS ANTIVIRAL.
SO WHEN ALL THE CELLS ARE
INFECTED AND WE'RE NOT LOOKING
AT VIRAL CELL TO CELL SPREAD AND
IMMUNE SIGNALING, WE FOUND THAT
THEN A LITTLE VIRAL PROTEINS
BECOME LACTYLATED AND 1 OF THEM
HAD TO DO WITH IT WAS AGAIN
INHIBITING THE ABILITY OF THE
THE IDR OF VIRAL PROTEIN TO GO
SPONTANEOUS ACTIVITY A LIQUID
SEPARATION IN THE NUCLEUS TO GO
INTO REPLICATION COMPARTMENTS SO
IT CAN INHIBIT THE FUNCTION OF A
VIRAL PROTEIN.
SO LACTYALATION CAN AFFAIRS TEAM
LEADERRER PROTEIN FUNCTION IN
MANY DIFFERENT WAYS.
>> DID WE GET A CLARIFICATION.
>> THAT INDIVIDUAL WAS ALL SET
WITH THAT RESPONSE.
, GREAT.
>> ANY OTHER ONLINE QUESTIONS?
>> OH LET ME SAY THE CME CODE?
, THERE IT IS 57688.
AND ANY -- THERE'S RECEPTION YOU
CAN MENTION.
>> YEAH, THERE'S' RECEPTION
OUTSIDE IMMEDIATELY AFTERWARDS
AND LET'S THANK ILEANA FOR A
REALLY BEAUTIFUL TALK.
[ APPLAUSE ]