WELCOME.
WELCOME, EVERYBODY, TO THIS
ANNUAL EVENT AT OUR OWN
WOMEN SCIENTISTS.
MY NAME IS DORIS WU, I'M AN
INVESTIGATOR AT NIDCD.
I'LL BE THE MODERATOR FOR
TODAY'S LECTURE.
SO YOU CAN ENTER YOUR QUESTIONS
AT THE BOX BELOW AND YOUR
QUESTIONS WILL BE ANSWERED
DURING THE Q & A SESSION.
SO NOW I'D LIKE TO INTRODUCE OUR
NEWLY ELECTED CHAIRPERSON OF THE
WSA, DR. GIGI STORZ AT NICHD.
>> THANK YOU, DORIS, AND ALSO
THANK YOU, DORIS, FOR ALL THAT
YOU'VE DONE TO ORGANIZE THE 2024
LECTURE.
SO AS DOOR DORIS MENTIONED, I M
CURRENTLY SERVING AS CHAIR OF
THE NIH WOMENS SCIENTISTS
ADVISORS.
THIS GROUP HAS BEEN AROUND FOR
OVER 30 YEARS NOW ADVOCATING AND
PROMOTING WOMEN SCIENTISTS AN
THE RESEARCH THAT THEY ARE
CONDUCTING.
ONE OF THE MOST IMPORTANT
EEFNLTS THAT IS CARRIED OUT
EVERY YEAR IS THIS ANITA ROBERTS
LECTURE, AND I THINK IT'S
APPROPRIATE TO SAY A FEW WORDS
ABOUT DR. ROBERTS.
SO DR. ROBERTS CAME TO NIH IN
1976 AND SPENT 30 YEARS AT NCI,
RISING TO THE POSITION OF CHIEF
OF THE LABORATORY OF CELL
REGULATION AND CARCINOGENESIS AT
A TIME WHEN NOT MANY WOMEN WERE
THE CHIEFS OF LABORATORIES.
SHE DID REALLY SEMINAL WORK
FOCUSED PRIMARILY ON TGF BETA
AND ITS ROLE IN THE GROWTH OF
EPITHELIAL AND LYMPHOID CELLS.
AND FOR QUITE A LONG PERIOD, SHE
WAS ONE OF THE 50 MOST CITED
SCIENTISTS.
BUT IN ADDITION TO REALLY HAVING
HER GROUP CONDUCT THIS
GROUNDBREAKING WORK, ANITA WAS
ALSO A SUPERSTAR MENTOR AND WAS
REALLY RECOGNIZED FOR HER
MENTORING TALENT, AND ALSO HER
ABILITY TO BALANCE HER FAMILY
AND PROMOTE HER FAMILY AND HER
WORK LIFE.
SO IT IS FOR THESE
ACCOMPLISHMENTS THAT DR. ROBERTS
IS CELEBRATED WITH THE ANITA
ROBERTS LECTURE.
I ALSO WANT TO POINT OUT THAT
ONE OTHER RECOGNITION OF
DR. ROBERTS' CONTRIBUTION CAN BE
FOUND ON THE NIH CAMPUS.
THIS IS ACTUALLY ONE OF MY
FAVORITE PLACES ON THE NIH
CAMPUS, AND THAT IS A FROG IN
HONOR OF DR. ROBERTS.
THIS FROG IS SITTING OUTSIDE OF
BUILDING 35, OUTSIDE THE --
BUILDING 37, OUTSIDE OF THE
BUILDING WHERE DR. ROBERTS USED
TO WORK, AND IT'S FUN TO SEE
EVERY ONCE IN A WHILE THE FROG
GETS A NEW PIECE OF CLOTHING AND
MOST RECENTLY, IT GOT DRESSED UP
FOR HALLOWEEN.
SO IN ADDITION TO REMEMBERS
DR. ANITA ROBERTS WITH THIS
FROG, THERE IS ALSO THIS LECTURE
SERIES, WHICH WAS STARTED IN
2006 WITH DR. ELIZABETH NABEL.
TODAY, WE WILL HAVE THE 25TH
LECTURE IN THIS SERIES WITH
DR. CAROLINA BARILLAS, AND
WITHOUT FURTHER ADO, I'M GOING
TO HAVE TWO REPRESENTATIVES OF
THE WOMEN'S FELLOW COMMITTEE
ACTUALLY INTRODUCE TODAY'S
SPEAKER, AND THOSE ARE
DR. CARRIE LONG AND DR. MUNA
FUYAL.
CARRIE, DO YOU WANT TO TAKE
OVER?
>> GOOD AFTERNOON, EVERYONE.
TODAY WE HAVE THE PRIVILEGE OF
WELCOMING A VISIONARY LEADER TO
THE STAGE.
FOR THE 2024 ANITA ROBERTS
LECTURE, DR. CAROLINA
BARILLAS-MURY WILL BE PRESENTING
AN IMPRESSIVE BODY OF WORK.
SHE RECEIVED A PH.D. IN
CHEMISTRY FROM THE UNIVERSITY OF
ARIZONA TUCSON.
IN 2003, DR. BARILLAS-MURY BEGAN
HER TENURE AS AN INVESTIGATOR IN
THE LABORATORY OF MALARIA AND
VECTOR RESEARCH.
THIS IS WITHIN THE NATIONAL
INSTITUTE OF ALLERGY AND
INFECTIOUS DISEASE.
SHE HAS SERVED AS HEAD OF THE
NIH MALARIA RESEARCH PROGRAM AND
IS CURRENTLY BOTH A
DISTINGUISHED INVESTIGATOR AND
IS A LABORATORY CHIEF OF THE
LNBR.
DR. BARILLAS-MURY HAS AUTHORED
124 PEER REVIEWED PUBLICATIONS
HIGHLIGHTING HER EMINENCE IN THE
FIELD OF MALARIA RESEARCH AND
TROPICAL MEDICINE.
HER ACCOLADES INCLUDE BUT ARE
NOT LIMITED TO BEING ELECTED AS
A FELLOW TO BOTH THE NATIONAL
ACADEMIES OF MED SUN AND SCIENCE
AND WINNING THE SANOFI PASTEUR
AWARD.
NOW I'LL HAND IT OFF TO MUNA.
>> THANKS, CARRIE, HELLO,
EVERYONE.
TODAY WE ARE GATHERED NOT ONLY
TO CELEBRATE DR. BARILLAS-MURY'S
SCIENTIFIC CONTRIBUTIONS BUT
ALSO IN THE SPIRIT OF ANITA
ROBERTS TO LAUDE HER EXCELLENT
MENTORSHIP.
DR. BARILLAS-MURY HAS TRAINED
OVER 50 YOUNG SCIENTISTS
INCLUDING POSTBAC, GRADUATE
STUDENTS AND POSTDOCTORAL
FELLOWS.
THESE TRAINEES HAVE GONE ON TO
HAVE SUCCESSFUL CAREERS IN
ACADEMIA, INDUSTRY, AND
GOVERNMENT ROLES, BOTH WITHIN
THE U.S. AND INTERNATIONALLY.
IN RECOGNITION OF THE POSITIVE
IMPACT ON THE PROFESSIONAL
DEVELOPMENT OF POSTDOCTORAL
FELLOWS, SHE WAS AWARDED THE
2013 OUTSTANDING MENTOR AWARD.
SHE ALSO MENTORS A GROUP OF FIVE
TENURE TRACK INVESTIGATORS WITH
THE AIM OF BUILDING A MORE
INCLUSIVE COMMUNITY WITHIN THE
INTRAMURAL RESEARCH PROGRAM.
IT WAS NOT A SURPRISE TO ME WHEN
I REACHED OUT TO HER PAST
TRAINEES ABOUT HER MENTORSHIP
AND THEY GOT BACK WITH THE
WARMEST REPORTS ON HER.
I'M GOING TO SHARE SOME OF THEIR
WORDS HERE.
DR. CAROLINA BARILLAS-MURY IS AN
EXCEPTIONAL MENTOR AND
SCIENTIST, ADMIRED FOR HUR
UNWAVERING DEDICATION TO
SCIENTIFIC EXCELLENCE AND HER
GENERGENUINE COMMITMENT TO HER
TRAINEES' GROWTH AND SUCCESS.
SEEING THE SIGNIFICANT TIME AND
EFFORT INTO UNDERSTANDING OUR
INDIVIDUAL ASPIRATIONS AND
PROVIDES GUIDANCE AND SUPPORT.
WHETHER IT IS WRITING
RECOMMENDATION LETTERS,
CONNECTING US WITH THE KEY
CONTACTS IN THE FIELD, OR
PROVIDING STRATEGIC ADVICE FOR
OUR CAREER DEVELOPMENT, SHE IS
ALWAYS THERE TO HELP US NAVIGATE
OUR PATHS TO SUCCESS.
I JOINED CAROLINA'S LAB WHEN SHE
FIRST STARTED AT THE NIH AND WAS
HER FIRST PH.D. STUDENT.
I HAD THE PRIVILEGE OF
WITNESSING FIRSTHAND HER
EXTRAORDINARY COMMITMENT TO
SCIENTIFIC EXCELLENCE, HER
INNOVATIVE APPROACH TO RESEARCH
AND HER UNWAVERING DEDICATION TO
THE GROWTH AND DEVELOPMENT OF
HER RESEARCH GROUP MEMBERS.
SHE CONSISTENTLY CHALLENGED ME
TO THINK CRITICALLY AND
CREATIVELY.
HER PASSION FOR SCIENCE AND HER
SENSE OF HUMOR ARE INFECTIOUS,
AND IT HAS INSTILLED IN ME A
DEEP SENSE OF PURPOSE AND
ENTHUSIASM FOR MY RESEARCH.
CAROLINA IS AN EXCELLENT MENTOR
AND SCIENTIST, DISTINGUISHED BY
HER UNWAVERING SCIENTIFIC RIGOR
AND EXEMPLARY WORK ETHIC.
MY TIME IN CAROLINA'S LAB WAS
PIVOTAL IN SECURING A PERMANENT
POSITION.
HER MENTORSHIP LEFT A LASTING
IMPACT ON MY CRITICAL THINKING
SKILLS, ENABLING ME TO POSE
SCIENTIFIC QUESTIONS WITH A
LEVEL OF DEPTH AND PRECISION I
WOULD NOT HAVE ACHIEVED
OTHERWISE.
DR. CAROLINA BARILLAS-MURY IS A
BEACON OF SCIENTIFIC INTEGRITY
AND MENTORSHIP.
WITHOUT HER KIND GUIDANCE AND
MORAL SUPPORT, I COULD NOT GIVE
THE FAMOUS PAPER ON TIME BOMB
THEORY, AND THE TENURE TRACK
FACULTY AT MY CURRENT INSTITUTE.
HER COMMITMENT TO ADVANCING
SCIENCE WHILE NURTURING THE NEXT
GENERATION OF SCIENTISTS
INCLUDING MYSELF IS TRULY
INSPIRING.
HER WILLINGNESS TO INVEST TIME
AND EFFORT INTO OUR GROWTH HAS
HAD A LASTING IMPACT ON ALL OF
US.
I STILL REMEMBER WHEN SHE USED
TO SING AND DANCE IN THE LAB
WHEN WE GOT GREAT DATA AND
DISCOVERY, AND I DO THE SAME
WITH MY STUDENTS IN KOREA.
A FEW OF THE THINGS I ADMIRE
MOST ABOUT CAROLINA ARE HER
UNPARALLELED SCIENTIFIC
KNOWLEDGE COMBINED WITH GREAT
SCIENTIFIC VISION.
HER ABILITY TO SEE BEYOND THE
IMMEDIATE ROADBLOCKS AND COME UP
WITH INNOVATIVE SOLUTION HAS
PIONEERED NEW APPROACHES TO
ENHANCE OUR UNDERSTANDING OF
INTERACTIONS.
ANOTHER ASPECT I ADMIRE MOST
ABOUT CAROLINA IS HER REMARKABLE
HUMILITY.
DESPITE HER NUMEROUS
ACHIEVEMENTS AND RECOGNITIONS,
SHE REMAINS APPROACHABLE AND
GENUINELY INTERESTED IN GIVING
ADVICE ON ANY TOPIC OF
DISCUSSION.
ALWAYS OFFERING GUIDANCE WITH
PATIENCE AND KINDNESS.
THANK YOU, CAROLINA, FOR YOUR
GUIDANCE AND SUPPORT, AND FOR
BEING AN INSPIRING MENTOR TO ALL
OF US.
THESE WORDS HIGHLIGHT HOW
DR. BARILLAS-MURY EXEMPLIFIES
MANY OF THE SAME QUALITIES THAT
MADE DR. ANITA B. ROBERTS SUCH A
GREAT SCIENTIST AND MENTOR.
IT IS A PLEASURE AND HONOR TO
WELCOME DR. CAROLINA
BARILLAS-MURY TODAY TO PRESENT
THE 2024 AKN ANITA B. ROBERTS
LECTURE.
ALL YOURS, CAROLINA.
>> THANK YOU.
THANK YOU VERY MUCH.
I WOULD LIKE TO START BY
THANKING THE ORGANIZERS AND THE
WOMEN SCIENTIST ADVISORS FOR THE
OPPORTUNITY TO GIVE THIS
LECTURE, I'M DEEPLY HONORED, AND
ALSO I'M OVERWHELMED WITH
EMOTION FROM THE COMMENTS
RECEIVED BY THE PERSONS THAT
HAVE BEEN IN MY LAB.
SO I WOULD LIKE TO START BY
SHARING THE SCREEN.
PUT THE SLIDES ON.
CAN EVERYONE SEE MY SLIDES?
I JUST WANT TO MAKE SURE.
WE'RE GOOD?
>> YOU ARE GOOD.
>> OKAY, GOOD.
SO GOOD MORNING, EVERYBODY.
MY LAB HAS BEEN WORKING FOR MANY
YEARS IN TRYING TO UNDERSTAND --
SO MY LAB HAS BEEN INTERESTED
FOR MANY YEARS IN UNDERSTANDING
THE MOSQUITO-PARASITE
INTERACTIONS THAT AFFECT MALARIA
TRANSMISSION WITH THE ULTIMATE
GOAL OF FINDING WAYS OF
DISRUPTING THE TRANSMISSION
CYCLE AND THEN PREVENT HUMAN
DISEASE.
WE WORK WITH THE ANOPHELES
GAMBIAE BECAUSE IT'S A MAJOR
VECTOR OF MALARIA IN AFRICA, AND
THIS MOSQUITO IS EXTREMELY
EFFICIENT IN TRANSMITTING THE
DISEASE FROM ONE PERSON TO THE
OTHER AND IT'S ONE OF THE
REASONS WHY MALARIA, ANOPHELES
GAMBIAE MALARIA, IS SUCH A
DEVASTATING DISEASE,
PARTICULARLY IN AFRICA.
AND SO TODAY, I'M GOING TO SHARE
WITH YOU ONE MAJOR PROJECT IN MY
LAB, ONE OF THE MAIN PROJECTS,
AND IT TRYING TO UNDERSTAND WHY
IS THIS TRANSMISSION OF DISEASE
SO EFFECTIVE?
SO AT THE BEGINNING, I WILL
SUMMARIZE HOW MOSQUITOES BECOME
INFECTED WITH PLASMODIUM AND
THEN WHAT KIND OF IMMUNE
RESPONSES THEY MOUNT TO THE
EARLY STAGES OF THE DISEASE.
SO WE HAVE ON TOP THIS HAS TAKEN
A BLOOD MEAL AND THE MOSQUITOES
BECOME INFECTED WHEN THEY INGEST
BLOOD FROM A VERTEBRATE HOST
THAT CONTAIN GAMETOCYTES.
THEY EMERGE FROM THE RED BLOOD
CELLS AND MATURE INTO GR GAMETE,
FERTATFERTILIZATION TAKES PLACED
THE FOCUS TODAY IS THIS STAGE,
THE BANANA SHAPED PARASITE, THE
PARASITES THAT ARE SUCCESSFUL GO
ON TO FORM AN OWE OOCYST, AND
TWO WEEKS LATER IT WILL RELEASE
THOUSANDS OF PAR SITES.
THIS MOSQUITO OPENS CIRCULATORY
SYSTEM, THEN WHEN THE MOSQUITO
THAT IS INFECTED WITH THIS BITES
THE NEXT PERSON, IT WILL
TRANSMIT THE DISEASE.
SO TODAY WE'RE GOING TO FOCUS ON
THIS PART OF THE SQUARE, WHAT
HAPPENS IN THESE EARLY STAGES.
SO I WANT TO POINT OUT THAT MOST
OF WHAT WE UNDERSTAND RIGHT NOW
ABOUT THIS RESPONSE, THE INITIAL
WORK, CAME FROM USING PLASMODIUM
VERSION OF MALARIA AND THEY --
THEY BREAK THE BARRIERS THAT
NORMALLY PREVENT THE GUT
MICROBIOTA FROM COMING IN DIRECT
CONTACT WITH EPITHELIAL CELLS
AND THEY CAUSE EA IRREVERSIBLE
DAMAGE TO THE CELL THEY INVADE.
THIS CELL DIES, BUT IT DIES
ASKING FOR HELP AND WHAT DO WE
MEAN BY THAT?
WHEN THIS CELL IS DYING, IT
ACTIVATES -- THIS ACTIVATES
CAASPP -- RESULTING IN THE
INDUCTION OF ENZYMES THAT
TRIGGER VERY STRONG NIGH
TRAITION RESPONSE.
NITRATION
RESPONSE.
PGE2 ATTRACTS HEMOCYTES, THEY'RE
IN THE IMMUNE CELLS, THEY COME
AND START PATROLLING THE BASAL
LAMINA.
THEY RELEASE MICRO VESICLES
LOCALLY AND WE HAVE SHOWN THAT
THIS RELEASE OF MICROVESICLES IS
ESSENTIAL FOR EFFECTIVE
ACTIVATION OF THE MOSQUITO
COMPLEMENT SYSTEM.
THE MOSQUITO COMPLEMENT SYSTEM
HAS A PROTEIN TEP 1.
THIS IS A FINAL -- OF THE
PATHWAY VERY SIMILAR TO FACTOR
C3 OF COMPLEMENT IN VERTEBRATES,
AND WHEN ACTIVATED BY SOME
FACTOR THAT IS NOT KNOWN YET
PRESENT IN THE MICROVESICLE,
THEN TEP1 DEPOSITS ON THE
SURFACE OF THE PARASITE AND
DESTROYS THE PARASITE.
OKAY.
SO HOW IMPORTANT IS THIS
TEP1 MEDIATED KILLING? THIS WAS
FIRST DESCRIBED BY A POSTDOC IN
THE LAB WHERE I WILL DID MY
POSTDOC, AND THE WAY THEY CAN
SEE IF SOME FACTOR IN THE
MOSQUITO IS IMPORTANT TO CONTAIN
THE INFECTION, YOU ASK WHAT
WOULD HAPPEN IF THAT GENE WAS NO
LONGER THERE.
MOSQUITOES, WE HAVE ONE TOOL
THAT IS VERY USEFUL, WE CAN
INJECTION STRANDED RNA GREATLY
DECREASES EX-PEGS OF THE GENES
THAT YOU WANT TO STUDY.
SO IN THIS EXPERIMENT, EACH DOT
REPRESENTS THE NUMBER OF
PARASITES IN ONE MOSQUITO.
SO THE NUMBER OF OOSEISTS THAT .
WE HAVE THE CONTROL GROUP
INJECTED WITH DOUBLE STRANDED
LACZ THAT SHOULD HAVE A
SILENCING EFFECT.
WHEN WE SILENCE TEP1 WE CAN SEE
INFECTION INCREASES, TYPICALLY
IT INCREASES ABOUT FIVE FOLD
WHEN TEP1 IS NO LONGER THERE.
SO THIS TELLS US THAT ABOUT 80
TO 85% OF PARASITES ARE
ELIMINATED BY THIS SYSTEM.
EVEN WHEN INFECTIONS ARE VERY
HIGH.
SO I'VE ONLY BEEN TALKING FOR A
COUPLE OF MINUTES AND THERE'S
ALREADY A CONTRADICTION IN WHAT
I'M TELLING YOU.
ONE, THAT THE MALARIA
TRANSMISSION IS EXTREMELY
EFFICIENT.
NOW I'M TELLING YOU THAT THE
MOSQUITO HAS A SYSTEM THAT CAN
ELIMINATE 85% OF THE PARASITES.
SO HOW IS THAT POSSIBLE?
SO FOR THE REST OF THE LECTURE,
I WILL HOPE TO CONVINCE YOU THAT
THIS IS BECAUSE PLASMODIUM
FALCIPARUM IS NOT A NORMAL
PARASITE.
IT HAS A MAGIC TRICK.
IT HAS A GENOME ON ITS SURFACE
THAT ALLOWS THIS TO BECOME
UNDETECTED BY THE MOSQUITO
IMMUNE SYSTEM SO IT CANNOT BE
DESTROYED BY THE COMPLEMENT
SYSTEM.
SO THIS WORK FOR PLASMODIUM
FALCIPARUM WAS DONE -- BUT NOW
INFECT IT WITH A HUMAN MALARIA,
PLASMODIUM FALCIPARUM, NOTHING
HAPPENED.
SO IT'S NOT KILLING THE
PARASITE.
WHEN THESE RESULTS CAME OUT IT
CAUSED A BIG UPROAR IN THE
FIELD, SOME INTERPRETED THAT
THIS WAS AN EXAMPLE OF A BIG
WASTE OF TIME, WHY USE THE MOUSE
MODEL, INVEST ALL THIS TIME IN
IMMUNE RESPONSE THAT IS
IRRELEVANT TO HUMAN MALARIA.
WE TACK A DIFFERENT PATH OR I
INTERPRETED THINGS IN A
DIFFERENT WAY, THE SAME DATA.
MY APPROACH WAS OKAY, I THINK WE
WERE VERY LUCKY THAT WE DID IT
FIRST WITH P. BERGHEI. THEN YOU
START -- THE LITERATURE --
ESCAPE OR THERE'S SOME CASES
WHERE SOME MOSQUITOES CAN KILL
SOME -- PARASITES.
THIS BROUGHT US BACK TO SOME
STUDIES IN THE 80s DONE HERE
AT NIH, FRANCIS COLLINS AND --
IN WHICH THEY HAD SELECTED
GENETICALLY A STRAIN OF
MOSQUITOES CALLED THE REFACTORY
STRAIN THAT KILLS ALL THE
PARASITES PRETTY MUCH THAT
INFECT THEM WITH ANY PARASITE.
EVERYTHING LOOKS FINE, BUT AS
SOON AS THE -- COMES IN CONTACT
WITH THE MOSQUITO -- THE
PARASITE IS DAMAGE AND AFTER
IT'S KILLED IT'S COVERED WITH A
BLACK PIGMENT.
WE KNOW THIS IS -- RELATED
BECAUSE IF YOU SILENCE TEP1 IT
CAN RESCUE.
THIS KILLS ALSO ANY PARASITE, BY
THAT I MEAN MOUSE MALARIA, BIRD
MALARIA, ALMOST ALL PLASMODIUM
FALCIPARUM WITH ONE INTERESTING
EXCEPTION.
IF YOU INFECT IT WITH PLASMODIUM
FALCIPARUM FROM A PATIENT FROM
WEST AFRICA, THE SAME PLACE
WHERE THIS MOSQUITO CAME FROM,
YOU SEE A VERY DIFFERENT PICT
PICTURE.
IN THIS GRAPH I WANT TO SHOW YOU
HOW DIFFERENT THIS IS.
BECAUSE WE CAN HAVE EITHER -- IN
A GIVEN MOSQUITO, IN THESE
GRAPHS WE HAVE THE PARASITES IN
THE X AXIS AND THE LIVE ONES IN
THE Y AXIS.
SO YOU SEE THAT IF THIS
MOSQUITO, THE REFRACTORY
MOSQUITO IS INFECTED WITH AN
AFRICAN PARASITE, THEN NONE OF
THE PARASITES ARE MELANIZED AND
THEY ALL SURVIVE.
IN CONTRAST, IF THIS CAME FROM
BRAZIL, A BRAZILIAN INFECTION,
AND THEN WHEN YOU LOOK THE A
THIS, IN THIS CASE, 9 7% ARE
MENONI SOO ED AND ABOUT 3%
SURVIVE.
SO THIS OPENS THE POSSIBILITY OF
THE DEVELOPING GENETIC STRATEGY
TO IDENTIFY THE PLASMODIUM
FALCIPARUM INVISIBILITY GENE.
SO WHAT IS DIFFERENT, WHAT DOES
THE AFRICAN PARASITE HAVE THAT
ALLOWS TO NOT BE DETECTED AND
NOT BE DESTROYED BY THE SYSTEM
THAT DESTROYS ALMOST ANY OTHER
PARASITE?
SO WE TOOK ADVANTAGE THAT --
WITH THESE TWO LINES THAT I SHOW
YOU, 78 FROM BRAZIL AND GB4 FROM
GHANA.
THIS ALLOWS US TO IDENTIFY PFS47
EXPRESSED ON THE SURFACE.
SO THEN WE -- ANOTHER LAB IN THE
NETHERLANDS HAD GENERATED A
KNOCKOUT PARASITE FOR PFS47,
SO -- FOR OTHER STUDIES, SO WE
MADE A COLLABORATION WITH THEM,
THEY SENT US THE PARASITE, AND
THIS IS WHAT THE NF54 WILD TYPE
PARASITE LOOKS LIKE IN THE
REFRACTORY STRAIN.
93% OF THE PARASITES SURVIVE AND
7% ARE MEL NIEZED.
BUT IF YOU DISRUPT NF54, YOU
HAVE A KNOCKOUT AND THIS LOOKS
VERY DIFFERENT AND 99% OF THE
PARASITES ARE DESTROYED.
IF YOU TAKE THESE KNOCKOUT
PARASITES BUT NOW YOU SILENCE
TEP, YOU CAN COMPLETELY REVERT
ALL THE PARASITES SURVIVE, NO
ONE IS MEL NIEZED.
THIS TELLS THE PARASITE NEEDS
THIS GENE TO EVADE BEING KILLED
BY THE MOSQUITO IMMUNE SYSTEM.
EVERYTHING ELSE APPEARS TO BE
NORMAL.
SO THIS GENE REALLY IS VERY
IMPORTANT FOR THIS.
SO IN SUMMARY FROM WHAT WE'VE
SAID SO FAR, SO PARASITES THAT
LACK PFS47 OR THEY HAVE THE
WRONG PFS47, FOR EXAMPLE, THE
BRAZILIAN PFS47 IN AN AFRICAN
MOSQUITO, THEY ACTIVATE NOS
CASPASE AND WE SEE THIS CAN
ACTIVATE CASPASE, AND THEN THERE
IS TEP 1 MEDIATED CLEARING.
HOWEVER, IF THE PARASITE HAS THE
RIGHT TYPE, THE AFRICAN
PARASITES WITH THE AFRICAN
MOSQUITO, THEN THERE'S INVASION
OF -- BUT NO ACTIVATION OF
CASPASE.
SO WHAT WE SEE HERE IS THE
PARASITE IS DYING BUT -- THE
CELL IS DYING BUT THE CELL DIES
WITHOUT BEING ABLE TO ASK FOR
HELP.
IT REALLY CANNOT NITRATE, AND IN
THE ABSENCE OF THAT, THEN THINGS
DON'T MOVE FORWARD.
SO THAT WAS THE WORKING MODEL.
SO WE CONCLUDED PLASMODIUM
FALCIPARUM -- IMMUNE SYSTEM --
THE FIRST STEP IN THIS PROCESS.
SO YOU SEE THIS IS THE SAME
GRAPH I SHOW YOU, YOU SEE HOW
DRASTICALLY DIFFERENT THESE
PHENOTYPES ARE, AND ALTHOUGH
THEY'RE SO DIFFERENT WHEN YOU
LOOK AT THE PROTEIN, PFS47
PROTEIN BETWEEN THESE TWO
STRAINS, THEY'RE VERY SIMILAR.
THEY'RE ALMOST IDENTICAL EXCEPT
FOR FOUR AMINO ACIDS HERE IN THE
CENTRAL DOMAIN, BETWEEN THESE
TWO, THERE'S FOUR AMINO ACIDS.
WE DID A SERIES OF EXPERIMENTS
WHERE WE TOOK ONE AMINO ACID AT
A TIME, WE TOOK THE GB4 AND
CHANGED THIS AMINO ACID FOR ONE
PERSON IN THE BRAZILIAN PARASITE
ONE AT A TIME, AND THEN WE FEED
THIS TO MOSQUITOES AND
PHENOTYPE.
AND WHAT WE FOUND IS THAT IF WE
CHANGE ANY AMINO ACID IN THE
AFRICAN STRAIN FOR A BRAZILIAN
ONE, NOW THE TRICK DOESN'T WORK,
THE PARASITE IS RECOGNIZED AND
DESTROYED.
SO FROM THESE, WE CONCLUDED THAT
THE MOLECULAR INTERACTIONS
BETWEEN PFS47 AND THE MOSQUITO
VECTOR NEED TO BE VERY PRECISE
AT LEAST IN SOME VECTOR PARASITE
COMBINATIONS, AND LEAST IN THIS
REFRACTORY STRAIN.
ALSO WITH USE OF STUDIES, TRYING
TO SEE IF ANTIBODIES TO SOME
REGION OF THIS PROTEIN COULD BE
PROTECTIVE, WE FOUND THAT DOMAIN
1 AND 3 ARE VERY NEUROGENIC
BUT -- DO NOT HAVE ANY EFFECT.
HOWEVER, THERE'S A REGION SHOWN
HERE IN RED IN DOMAIN 2 WHERE
ANTIBODY BINDING POLYCLONAL
OR -- ANTIBODIES GREATLY REDUCE
TRANSMISSION.
SO YOU ADD EITHER IGG OR YOUR
SPECIFIC IGG TO PFS47 -- IGG OF
PFS47 IN IMMUNIZED MICE AND WE
FIND IT REDUCES THE INFECTION,
BOTH IN -- AND T ANOPHELES GAMB-
OKAY, SO NOW LET'S GET OUT OF
THE LAB AND THINK OKAY, THAT'S
ALL IN THE LAB, AND NOW WHAT ARE
THE CONSEQUENCES OF THESE
INTERACTIONS IN THE WORLD?
HOW IMPORTANT HAS THIS ABILITY
TO EVADE THE MOSQUITO IMMUNE
SYSTEM, HOW IMPORTANT IS THIS
FOR MALARIA TRANSMISSION AROUND
THE WORLD?
TO ANSWER THIS QUESTION THEN, WE
TOOK A GENETIC APPROACH.
SO WE ANALYZE -- SEVERAL YEARS
AGO -- THE FIRST 364 SEQUENCES
AVAILABLE OF PLASMODIUM
FALCIPARUM FROM DIFFERENT HUMAN
ISOLATES AROUND THE WORLD, AND
THEN WHEN WE ANALYZED IT, WE
FOUND 42 DIFFERENT HAPLOTYPES,
AND WE DEFINE A HAPLOTYPE OF ANY
AMINO ACID DIFFERENCE BETWEEN
TWO PROTEINS.
WHEN WE DID A PHYLOGENETIC TREE,
WE CAN SEE IN GROUPS WE HAVE THE
ORANGE GROUP, THE BLUE AND THE
GREEN.
OF COURSE THEY'RE CLOSER, THE
THE ONES THAT ARE MORE SIMILAR
ARE CLOSER TO EACH OTHER.
WHEN WE PUT THESE IN A MAP, WE
SEE THE DISTRIBUTION OF THESE
COLORS IS NOT RANDOM.
-- IN SOUTH AMERICA ARE BLUE,
ORANGE IN AFRICA, AND GREEN
DOMINATES IN ASIA.
IN NEW GUINEA WE HAVE MOSTLY
ASIAN BUT YOU FIND SOME OTHER
HAPLOTYPES TO THIS REGION AND
FROM OTHER CONTINENTS.
OKAY.
BUT THINKING ABOUT MALARIA, THIS
IS VERY UNUSUAL BECAUSE WHY
WOULD YOU HAVE GEOGRAPHIC
DIFFERENCES, RIGHT?
YOU HAVE HUMANS, WE ARE VERY
SIMILAR, TO HAVE A HOST, THEN WE
HAVE A SINGLE PARASITE, BUT WHAT
IS VERY DIFFERENT IS THE
MOSQUITOES.
TO GIVE YOU AN IDEA, THIS IS A
SIMPLIFIED MAP, WHAT YOU NEED TO
REMEMBER FROM THIS MAP IS THAT
ALL THESE COLORS REPRESENT THE
DIFFERENT MOSQUITOES THAT ARE
TRANSMITTING MALARIA IN
DIFFERENT AREAS OF THE WORLD,
AND THERE'S MORE THAN 40 MAJOR
VECTORS OF MALARIA IN DIFFERENT
PLACES.
ANOTHER ASPECT TO REMEMBER IS
SOME MOSQUITOES ARE
EVOLUTIONARILY VERY DISTANT.
FOR INSTANCE THOSE IN SOUTH
AMERICA AND AFRICA SEPARATED
100 MILLION YEARS AGO, WHEN
THESE TWO CONTINENTS SEPARATED,
THEN THESE TWO, AND OFTEN
IT'S -- IN AFRICA BEGAN TO
DIVERGE.
SO BASICALLY THERE'S DIFFERENT
MOSQUITOES TRANSMITTING IN
DIFFERENT PLACES AND SOME OF
THEM ARE EVE LEUT EVOLUTIONARILY
DISTANT.
SO DETERMINED BY PARASITE
SELECTION BY THE MOSQUITO
VECTORS.
OKAY.
SO HOW DO WE SHOW THAT?
SO IN ORDER TO TEST THIS
HYPOTHESIS, THIS IS VERY HARD TO
DO IN THE FIELD BECAUSE YOU CAN
MOVE MOSQUITOES AROUND, YOU
DON'T WANT TO BE FAMOUS AND
BRING AFRICAN MOSQUITOES IN
OTHER PLACES AND CAUSE A
PANDEMIC, SO WE BROUGHT THIS TO
THE LAB.
SO WE EXPANDED THE COLONIES TO
HAVE AT THE SAME TIME, WE WOULD
INFECT THE THREE MOSQUITOES AT
THE SAME DAY, AT THE SAME TIME
WITH PARASITES FROM DIFFERENT
AREAS.
SHOWN HERE BY THE AS TR ASTERISM
AFRICA, AMERICA OR ASIA.
SO WHEN WE DID THESE EXPERIMENTS
WE FOUND SOME VERY STRIKING
RESULTS.
SO IN THE FIRST ONE, YOU TAKE AN
AFRICAN PARASITE, THIS IS NF54,
INFECTING ANOPHELES GAMBIAE FROM
AFRICA, VIRUS FROM ASIA OR THE
AMERICAS, AND WHAT YOU SEE IS
THAT THE AFRICAN PARASITES
INFECT THE AFRICAN MOSQUITOES
BETTER THAN THOSE OF OTHER
CONTINENTS.
SO NOW WHAT WOULD HAPPEN IF WE
TAKE THE SAME MOSQUITOES BUT NOW
THE PARASITE CAME FROM ASIA.
IF YOU DO THAT, THIS IS FROM A
PATIENT FROM ASIA, NOW THIS
ASIAN PARASITE INFECT THE
MOSQUITOES -- IN THE NEW WORLD
THEY REALLY DON'T DO WELL.
WHAT IF THIS WAS A PARASITE THAT
CAME FROM SOUTH AMERICA, FROM
BRAZIL?
THEN THIS ONE DOES BETTER IN
THIS AMERICAN VECTOR THAN WITH
THOSE FROM AFRICA OR -- SO THIS
TELLS US THERE IS SELECTION THAT
THE PARASITES IN A GIVEN
CONTINENT DO BETTER.
IF THEY'RE INFECTED WITH
MOSQUITOES FROM THE SAME PLACE,
THAT MAKES SENSE BECAUSE FOR
THE -- THEY HAD TO SURVIVE IN
THE VECTOR.
IT'S THE ONLY WAY THEY CAN MOVE
FROM ONE PERSON TO ANOTHER.
THEY MUST PASS THROUGH A
MOSQUITO.
SO NOW TO WHAT EXTENT IS THIS
SELECTION -- COULD BE BY MANY
FACTORS BUT DO THIS HAVE
ANYTHING TO DO WITH THE MOSQUITO
IMMUNE SYSTEM?
THE WAY WE CAN ADDRESS THAT IS
THAT TEP1 THAT I TALKED ABOUT
HAVE A STABILIZER THAT IS VERY
WELL CONSERVED AND THERE'S A
CLEAR OR THE LCLEAR ORTHO LOG SN
DISRUPT THE MOSQUITO IMMUNE
SYSTEM BY SILENCING THE
STABILIZER AND THEN ASK WHEN WE
DO THAT CAN WE REVERT -- SO WE
DID THOSE EXPERIMENTS AND I'LL
SHOW YOU SOME EXAMPLES.
THIS IS THE ASIAN MOSQUITO WITH
AN AFRICAN PARASITE.
NORMALLY IT DOESN'T DO SO WELL,
BUT IF YOU SILENCE LRIM1 THEN
INFECTIONS INCREASE
SIGNIFICANTLY.
NOW WE GO TO MOSQUITOES FROM THE
NEW WORLD, THIS IS ALBIMANUS
WITH AN AFRICAN PARASITE, REALLY
BAD, BUT IF YOU SILENCE, YOU GET
THIS STRIKING INCREASE.
AND THE SAME THING HAPPENS WITH
THE ASIAN PARASITES.
SO WHAT DOES THIS MEAN, THAT IF
YOU TURN OFF OR DISRUPT THE
MOSQUITO IMMUNE SYSTEM, ALL THE
PARASITES CAN INFECT ALL THE
DIFFERENT MOSQUITOES.
THIS TELLS US IS THE MOSQUITO
IMMUNE SYSTEM THE ONE EXERTING
THIS SELECTED PRESSURE.
WE DID SOME OTHER EXPERIMENTS
THAT I'M JUST GOING TO
SUMMARIZE.
WE ALSO TOOK THE N TB. 54
KNOCKOUT PARASITES AND REPLACED
THE PF47 FROM AFRICA -- FROM THE
NEW WORLD AND CHANGING THAT ALSO
CHANGED THE COMPATIBILITY.
SO BASED ON THIS, THE PICTURE
THAT WE HAVE IS THAT MALARIA IS
KNOWN TO HAVE ORIGINATED IN
AFRICA AND THE -- REGIONS IN
AFRICA, AND IT'S THOUGHT TO HAVE
BEEN A SINGLE TRANSFER FROM A
GORILLA, PARASITE, TO A HUMAN.
ONE THE INFECTIONCE THE INFECTIS
ESTABLISHED IN HUMANS, GORILLAS
GORILLAS -- SEVERAL CENTURIES
MOVED TO ASIA, BUT THE
MOSQUITOES WERE LEFT BEHIND, SO
NOW THESE PARASITES HAD TO ADAPT
TO A NEW MOSQUITO IN ORDER TO
ESTABLISH LOCAL CYCLES OF
TRANSMISSION.
THE OTHER ADAPTATION WAS FROM --
REGIONS TO ASIA AND THEN ANOTHER
ADAPTATION WITHIN AFRICA WAS
HUMAN CIVIC REGIONS WHERE IT
ORIGINATED TO THE REGION WHERE
IT ORIGINATED -- SO ALSO MORE
RECENTLY THIS HAPPENED OVER
THOUSANDS OF YEARS, BUT MORE
RECENTLY WITH THE DISCOVERY OF
THE NEW WORLD, IT'S ESTIMATED
ABOUT 8 TO 10 MILLION AFRICAN
SLAVES WERE BROUGHT TO THE
AMERICAS AND MANY OF THEM WERE
INFECTED WITH PLASMODIUM
FALCIPARUM.
THIS PARASITE NOW FINDS ITSELF
IN A DIFFERENT CONTINENT WITH
MOSQUITOES THAT WERE REALLY,
REALLY DIFFERENT, AND SO THIS
WAS ANOTHER BIG SELECTION
BECAUSE ONLY THOSE, MOST
PARASITES -- TODAY IN AFRICA
HAVE NEVER BEEN ABLE TO
ESTABLISH -- HAVE NOT BEEN ABLE
TO ESTABLISH IN THE NEW WORLD,
ONLY THE ONES THAT HAVE THOSE
FOUR AMINO ACID DIFFERENCES THAT
I SHOW YOU ARE ABLE TO BE
TRANSMITTED AND THAT SELECTION
IS STILL HAPPENING TODAY.
OKAY.
SO BASED ON THIS, NOW WE GO BACK
TO THE LAB, WE GET OUT OF THE
FIELD, WE GO BACK INTO THE LAB.
SO BASED ON THIS, WE PROPOSE A
MODEL THAT WE CALL THE LOCK AND
KEY THEORY.
WITH THESE, WE CAN THINK OF
PFS47 AS A KEY AND THERE'S
DIFFERENT VERSIONS OF THE KEY OR
HAPLOTYPES, AND THEN HERE
HAPLOTYPES 1, 2 AND 3, AND THIS
KEY INTERACTS WITH THE LOCK IN
THE MOSQUITO, A RECEPTOR, AND
THE INTERACTION OF THE KEY WITH
THE LOCK ALLOWS THE PARASITE TO
TURN OFF THE NITRATION RESPONSE
IN THE EPITHELIAL CELL.
THE KEY YOU NEED DEPENDS WHICH
LOCK YOU'RE TRYING TO OPEN, SO
WHAT WORKS GREAT FOR AFRICA
DOESN'T WORK SO WELL IN THE NEW
WORLD AND SO FORTH.
SO THERE HAS TO BE A MATCH
BETWEEN THE KEY AND THE LOCK FOR
THIS SYSTEM TO WORK.
SO THE KEY TO THIS THEORY IS TO
IDENTIFY WHAT IS THE LOCK, WHAT
IS THE MOSQUITO RECEPTOR OF
PFS47.
SO IN ORDER TO DO THAT, FIRST WE
MOBILIZED SOME -- WE TOOK --
THEN WE REMOVED THE CYTOPLASM
PART AND TOOK THE INSOLUBLE PART
THAT HAS THE MEMBRANES AND
PROTEINS.
WE RUN THOSE, WERE DISSECTED AND
CLEANED EITHER 24 OR 48 HOURS
AFTER FEEDING THEM ON HUMAN
SERUM, AND THEN WE TRANSFERRED
THESE PROTEINS TO A MEMBRANE AND
THEN THE MEMBRANE IS INCUBATED
WITH WHAT WE CALL A BAIT, WHICH
IS RECOMBINANT PFS47 AND THERE'S
NO BAIT CONTROL, AND THEN WE ASK
DID THIS PFS47, THIS RECOMBINANT
PROTEIN BIND TO ANY SPECIFIC
BAND AND WE FIND IT DOES.
WE CUT THE SPOT, DID A MASS SPEC
AND IDENTIFIED A PUTATIVE
RECEPTOR.
WE TOOK THIS PUTATIVE RECEPTOR,
WE SILENCED EXPRESSION WITH
DOUBLE STRANDED RNA AND WE SHOW
THAT THE SIGNALING IS GREATLY
REDUCED WHEN WE SILENCE THE
EXPRESSION.
TO MAKE SURE WE HAVE THE RIGHT
PROTEIN, WE ALSO EXPRESS REIMI
NENT RECEPTOR AND BINDING -- YOU
SEE PLASMA RESONANCE
RESPONSES -- AND THEN EITHER
WHEN WE IMMOBILIZE THE RECEPTOR
AND THEN BOUND SOLUBLE PFS47 OR
IMMOBILIZE PFS47 AND INCUBATE
WITH SOLUBLE RECEPTOR, IN BOTH
CASES WE GET HIGH AFFINITY
BINDING.
SO THIS IS A HIGH AFFINITY
INTERACTION.
WE ALSO MADE ANTIBODIES TO IT,
AND IN THIS PICTURE HERE ON THE
LEFT SIDE, WE CAN SEE IN RED,
THIS IS THE ACTIN IN THE MICRO
VILLI, THIS IS THE LUMEN SIDE OF
THE MOSQUITO --, HERE IS WHERE
WE SEE THE STRONGEST THINNING
FOR THE PFS47 RECEPTOR, AND
BOUNDARIES IN THE INTERCELLULAR
JUNCTIONS.
NOW ON THE RIGHT SIDE, WE HAVE A
PARASITE IN THE PROCESS OF --
THAT HAS JUST INVADED THIS --
CELL.
SEW WITH HAVE THE PARASITE IN
GREEN, HERE YOU SEE THIS RED
SPOT, THIS IS A RECEPTOR.
THIS IS LOOKING FROM THE TOP AND
THIS IS LACKING SIDEWAYS.
LOOKING SIDEWAYS.
AT THE POINT WHERE THE PAWR SITE
INVADES THE CELL, THERE'S
AGGREGATION OF THE RECEPTOR.
SO THERE'S CLEAR CLOSE
INTERACTION OF THE SURFACE OF
THE PARASITE WITH THIS RECEPTOR.
ALSO WHAT IS THIS PUTATIVE
RECEPTOR?
IT'S A DEEP PROTEIN THAT HAS NO
OR THE LOGS, NONE FUNCTIONING IN
ANY OTHER SYSTEM.
IT WAS CLEAR -- -- PROTEINS WITH
THESE -- DOMAINS HAVE STRUCTURAL
HOMOLOGY TO OTHER PROTEINS THAT
ARE INTERACTING PROTEINS, AND
ALSO SOME MORE RECENTLY --
CONTAINING PROTEINS HAVE BEEN
PROPOSED TO BE PATHOGEN
REGULATION RECEPTORS, IN OTHER
SYSTEMS.
SO HERE WE HAVE THE PREDICTED
STRUCTURE OF THIS RECEPTOR IN
ANOPHELES GAMBIAE, DIRUS AND
ALBIMANUS.
SO WHAT HAPPENS IF WE REDUCE
EXPRESSION OF THIS RECEPTOR?
THIS IS HARD TO DO BUT WHEN WE
DO THAT, WE SEE THERE'S A
SIGNIFICANT REDUCTION IN THE
LEVEL OF INFECTION.
SO THE INTERACTION OF PFS47 WITH
THIS RECEPTOR IS IMPORTANT FOR
THE PARASITE TO SURVIVE.
WE ALSO WANTED TO SEE MORE
DETAIL, WE HAD DETERMINED THAT
CASPASE ACTIVATION WAS IMPORTANT
BASED ON MRNA EXPRESSION AND
ON -- ACTIVITY BUT WE WANTED TO
SEE THE CELL BIOLOGY IN MORE
DETAIL.
SO WHAT IS THE DIFFERENCE OF HOW
A CELERY SPONDZ WHEN IT'S
INVADED BY EITHER WILD TYPE OR
KNOCKOUT PARASITE?
SO IN THE WILD TYPE -- LOOK
REALLY NICE WHEN THEY COMPLETE
INVASION, THEY LOOK LIKE THIS,
WHILE IN THE KNOCKOUT, THEY'RE
ALL FRAGMENTED.
THEY'RE TORN APART.
HERE'S THE QUANTITATION.
IN THE KNOCKOUT WE HAVE MORE --
PARASITES AND BECAUSE MORE OF
THEM DIE, WE SEE ALSO HIGHER
CASPASE ACTIVATION, AND WE
ALSO -- EITHER BY ENZYMATIC
ASSAY OR COUNTING WHAT IS THE
PERCENT OF INVADED CELLS THAT
ARE CASPASE H-POSITIVE.
TO GIVE YOU AN IDEA OF WHAT THIS
LOOKS LIKE IN THE MICROSCOPE, SO
IN THE KNOCKOUT, WE SEE HIGH
ACTIVATION OF CASPASE IN THE
CELL AND THIS IS AN ANTIBODY
THAT DETECTS ACTIVATED CASPASE.
SO WE HAVE A BIG DIFFERENCE IN
HOW THESE TWO SYSTEMS ARE
WORKING.
THIS ONE IS VERY ACTIVE, WHILE
IN THE WILD TYPE, THIS IS A
QUIET CELL ON WHICH NIGH
TRAITION AN CASPASE ACTIVATION
DON'T TAKE PLACE.
ALSO WHEN WE LOOK AT -- TO LOOK
AT DNA FRAGMENTATION, WE SEE A
LOT OF DNA FRAGMENTATION IN THE
KNOCKOUT BUT NOT IN THE WILD
TYPE.
OKAY.
HOW DOES THIS WORK?
WE HAVE THIS PROTEIN WITH NO --
TO ANYTHING, NO FUNCTION, SO HOW
IS THIS AFFECTING APOPTOSIS?
SO WE DECIDED OKAY, LET'S TRY TO
SEEK OR FIND A PARTNER THAT
INTERACTS THAT MAYBE CAN GIVE US
A HINT OF HOW THIS COULD WORK.
SO WE DID A SIMILAR EXPERIMENT
TO WHAT WE USED TO DETECT THE
RECEPTOR, BUT FIRST WE WANTED TO
SEE IN WHICH FRACTION IS MOST OF
THE RECEPTOR IN THE MOSQUITO
WHEN WITH HE DO THIS.
SO TO DO THIS WE TOOK AGAIN --
GOT A READ OF THE CYTOPLASMIC
FRACTION, THEN WE EXTRACTED --
TO REMOVE MEMBRANE AND THEN WE
HAVE INSOLUBLE PART.
HERE WE FIND THIS IS WHERE MOST
OF THE RECEPTOR IS.
SO THEN WE PREPARED THE
HOMOGENEOUS FROM THIS FRACTION
TO SEE -- SO WE PREPARED NEW
GELS, WE RUN THE GEL, THEN
TRANSFER THE PROTEIN TO A
MEMBRANE AND THEN WE PROBE THIS
WITH A P47 BAIT, MEANING WE AS
ASKED P47 RECEPTOR BIND ANYWHERE
AND WE FOUND THERE'S STRONG
BINDING TO PROTEINS AND
SOMETIMES WE SEE A SECOND
BINDING APPEARS TO BE THE
DIMER -- PROTEIN IS A DIMER.
WHEN WE WENT AHEAD AND RAN IT
THROUGH THE GEL AND -- PROTEIN
70-3, AND WHEN WE REDUCE -- WHEN
WE SIGN THIS EXPRESSION OF THIS
PROTEIN WE GREATLY REDUCE THE
SIGNAL IN THE FAR WEST.
THERE'S VERY LITTLE BINDING WITH
THE RECOMBINANT RECEPTOR WITH
THE MEMORY AND THIS IS THE
ACTING CONTROL TO SHOW THAT IT
WAS PROTEIN.
OKAY.
SO WE WANTED TO CONFIRM THESE
PROTEINS INTERACT SO WE MAKE THE
TWO PROTEINS RECOMBINANT SO IF
WE RUN THE P47 RECEPTOR IN A
MEMBRANE, THEN THE BAIT, THE
RECOMBINANT HSC70 PROTEIN BINDS
TO IT.
WE CAN ALSO DO BINDING
INTERACTIONS WITH THE LYSIS --
IN BOTH CASES WE GET SATURABLE
BINDING.
SO WHAT IS THE EFFECT OF
REDUCING EXPRESSION OF PROTEIN
IN WILD TYPE PLASMODIUM
FALCIPARUM.
IT'S ABLE TO INTERACT WITH THE
RECEPTOR, THEN THE RECEPTOR
INTERACTS WITH EACH PROTEIN.
SO IF YOU REMOVE IT FROM THE
SYSTEM, THIS SHOULD AFFECT THE
PARASITE, THE PARASITE SHOULD
NOT DO SO WELL BECAUSE WE'RE
SAYING THAT EACH PROTEIN IN THIS
INTERACTION IS IMPORTANT.
IT WILL BE AFFECTED WHEN WE
REDUCE EXPRESSION.
AND THIS IS INDEED WHAT WE SEE
WHEN WE SILENCE REDUCED
EXPRESSION -- WE BEGIN TO SEE
INCREASING FRAGMENTED PARASITES,
REDUCTION IN THE NUMBER OF LIVE
PARASITES, AND INCREASE IN --
ACTIVITY.
SO IF YOU -- THE PICTURE NEEDS
TO LOOK LIKE A KNOCKOUT
PARASITE, LIKE PFS47 IS NO
LONGER WORKING IF THE INITIAL
PROTEIN IS NOT THERE.
SO THERE'S -- THAT IS VERY
IMPORTANT TO KILL THE PARASITE,
SO WE ASKED IS THIS DEATH THAT
WE'RE SEEING WHEN WE ASSIGN EACH
PROTEIN MEDIATED BY CASPASE?
TO DO THAT EXPERIMENT, THIS IS A
REPEAT OF WHAT I JUST TOLD YOU,
THAT THE NUMBER OF LIVE
PARASITES REDUCES WHEN YOU --
PROTEIN 70 THIS IS IN WILD TYPE
PARASITES THAT HAVE PFS47.
IN THE WILD TYPE THIS SYSTEM IS
NO LONGER ACTIVE.
WHEN WE DO -- EACH OF PROTEIN
17 -- YOU CAN RESCUE THIS.
SO WHEN YOU RESCUE THIS, THIS IS
TELLING US THE KILLING IS
MEDIATED BY CASPASE -- 2.
SO ALL TOGETHER SUMMARY OF WHAT
WE'VE SAID SO FAR, WHAT WE KNOW
SO FAR HOW THIS IS WORKING, SO
PLASMODIUM FALCIPARUM HAS A
MAGIC PROTEIN CALLED PFS47 ON
ITS SURFACE, AND THIS PROTEIN
HAS DIFFERENT HAPLOTYPES, IT IND
ACTS WITH THE MOSQUITO RECEPTOR
THAT IS DIFFERENT IN DIFFERENT
MOSQUITOES, AND THIS RECEPTOR
INTERACTS WITH PROTEIN HSC70 AND
THROUGH SOME MECHANISM
SUPPRESSES CASPASE AND IT HAS
SHOWN IN OTHER SYSTEMS IN
VERTEBRATES THAT THERE'S SOME
EXAMPLES OF -- PROTEIN HAVING AN
ANTIPIPE TO TICK EFFECT, THE
PRECISE MECHANISM, WE STILL
DON'T KNOW.
-- MODELS FOR THESE THREE
PROTEINS, AND WE HAVE BEEN
TRYING FOR MANY YEARS TO
CRYSTALLIZE AND SOLVE THE
STRUCTURE F OF THESE PROTEIN, TS
HAS PROVED TO BE VERY
CHALLENGING.
VERY RECENTLY WE HAVE BEEN ABLE
TO PRODUCE LARGE AMOUNTS OF
PFS47 RECEPTOR OF EACH PROTEIN
AND WE ARE IN THE PROCESS OF
FORMING A STABLE DIMER, AND
BECAUSE -- PROTEIN IS BIG, THE
TWO TOGETHER HAVE ALMOST 100 --
AND THAT PUTS US IN A SIZE -- TO
SOLVE THE STRUCTURE.
OUR DREAM IS TO BE ABLE TO PUT
THESE THREE INTERACT, AND WHAT
ARE THE MOLECULAR INTERACTIONS.
THIS IS PARTICULARLY
IMPORTANT -- PFS47 AND THE
RECEPTOR BECAUSE THIS WILL BE A
MOLECULAR LANGUAGE OF
COMPATIBILITY.
WE KNOW THE SEQUENCES OF
PARASITES ALL OVER THE WORLD AND
WE KNOW THE SEQUENCES OF THE
RECEPTORS IN THE MOSQUITO, SO IF
WE KNEW HOW THEY INTERACT, WE
COULD PREDICT HOW LIKELY IS A
PARASITE TO BE TRANSMITTED BY A
GIVEN VECTOR IN A DIFFERENT
PLACE, AT LEAST IN TERMS OF THE
MOSQUITO ANALYSIS.
INDEED THIS -- DOMAIN 2, THERE'S
TWO OF THEM THAT ARE FIXED, TWO
NUCLEOTIDES -- COMBINATION THAT
ARE ALMOST FIXED IN DIFFERENT
CONTINENTS, PACEICALLY FOR THESE
TWO POSITIONS, 98% OF PARASITES
ARE T-T IN THE AMERICAS, C-C IN
AFRICA, AND T-C IN ASIA.
SO THIS ALLOWS US TO GENERATE --
TO DEVELOP A HIGH RESOLUTION
MELTING GENOTYPE ASSAY,
BASICALLY YOU RUN A PCR, YOU
MELT THE PRODUCTS AND THEN YOU
CAN TELL IF THE PARASITE --
WHERE IT'S LIKELY THAT IT CAME
FROM.
THIS IS VERY USEFUL BECAUSE NOW
WITH -- MANY PEOPLE ARE MOVING A
LOT, SO SOMETIMES YOU ENCOUNTER
PLACES WHERE MALARIA HAS BEEN
ERADICATED, YOU NEED TO KNOW
WHERE IT CAME FROM, WHAT ARE THE
CHANCE OF AN EPIDEMIC.
I THINK YOU HEARD IN MARYLAND,
EVERY YEAR THERE'S A FEW MALARIA
CASES, SOMETIMES CASES OF LOCAL
TRANSMISSION AND THEY REALLY
NEED TO KNOW WHERE THESE
PARASITES ARE COMING FROM, SO
THIS IS EXTREMELY HELPFUL FOR
THAT.
EVERYTHING I'VE TOLD YOU SO FAR
IS A MAJOR DETERMINANT -- VERY
HIGH MORTALITY, PARTICULARLY IN
CHILDREN.
P.VIVAX IS A MAJOR PROBLEM -- SO
THEY ARE RESISTANT TO INFECTION
WITH THE VIRUS.
SO WE ASK WAS THIS COMPATIBILITY
AND PV47 WHICH IS THE ORTHO LOG
IN VIRUS, WOULD THIS ALSO BE
GOING THROUGH A SIMILAR PROCESS?
SO HERE AGAIN ALL THE VAWR VARI,
THERE'S NOR GENETIC DIVERSITY,
THE HIGHEST LEVELS ARE IN ASIA
AND NEW GUINEA, AND MOST
PARASITES ARE SEGREGATED BY
CONTINENT.
ALTHOUGH THE PARASITE ORIGINATED
IN AFRICA, THERE'S LESS
DIVERSITY AND LESS PARASITES IN
AFRICA BECAUSE ALMOST EVERY --
MOST PEOPLE IN AFRICA RECEIVE
SOME INFECTION WITH VIVEX.
NEW CASES ARE APPEARING AND IT'S
NOT CLEAR WHERE EXACTLY THEY
CAME FROM.
NOW I IF WE LOOK AT THE
DIFFERENCES, THEY ARE NOT
RANDOM.
IN THIS THE PINK AREA IS THE
DOMAIN BETWEEN THE TWO SYSTEMS
THAT WE TALKED ABOUT THAT IS
ALSO VERY IMPORTANT, I WANT TO
CALL YOUR ATTENTION TO A
DIFFERENCE HERE CLOSE TO THE END
TERMINUS, AND HERE FOR
COMPARISON, WE HAVE PFS47 AND
AGAIN THE NUCLEOTIDES ARE HERE
IN DOMAIN 2, BETWEEN THESE TWO
SYSTEMS AND THAT'S TRUE HERE.
THIS IS JUST A NETWORK ANALYSIS
TO STUDY ALL THE HAPLOTYPES AND
HOW THEY'RE CONNECTED, THE
CLOSER THEY ARE, THE LESS
DISTANCE THERE ARE BETWEEN THEM.
AND WE CAN SEE THAT THE ONCE IN
THE NEW WORLD ARE MORE SEPARATED
FROM THE REST.
IF WE LOOK AT THIS IN A
QUANTITATIVE WAY WITH THIS
HEAT -- MAP, AND FST IS THE
HIGHEST, BETWEEN .6 AND .86
BETWEEN THE SOUTH AMERICAN
CONTINENT AND THE REST OF THE
WORLD, AND THIS IS REFLECTED
HERE.
TO GIVE YOU AN IDEA, IF IT MORE
THAN 7.05, WE'LL HAVE EXTREMELY
HIGH POPULATION DIVERSITY, THIS
SELECTION BY GEOGRAPHIC AREA AND
THAT WE THINK IS CREATED BY THE
MOSQUITOES.
THEN IN CONTRAST, THERE'S SOME
OTHER REGIONS LIKE THOSE IN
AFRICA AND NEW GUINEA ARE NOT SO
WELL DIFFERENTIATED, THERE'S
ACTUALLY SOME OVERLAP, SO THE
FST VALUE IS NOT SO HIGH.
LAST PART OF THE TALK, THIS IS
SOMETHING FASCINATING.
THIS IS PLASMODIUM VIVEX IN
MEXICO, TRANSMITTED BY TWO
DIFFERENT VECTORS.
IN THE HIGHLANDS IN GREEN AND
THEN THE COASTAL AREA IS THE
MAIN VECTOR.
THIS STUDY WAS DONE HERE IN THE
BORDER BETWEEN MEXICO IN THE
BORDER BETWEEN MEXICO AND
GUATEMALA.
SO IN THAT AREA, WE SEE SEVERAL
YEARS AGO, XIN ZHUAN SU IN
COLLABORATION WITH THE GROUP,
THEY DID A VERY INTERESTING
STUDY ON WHICH THEY GENOTYPED
PARASITES FROM THE HIGHLANDS AND
THOSE FROM THE COASTAL AREAS,
AND THEY FOUND THERE WERE THREE
GENOTYPES TOO IN THE HIGHLAND
AND ONE MAJOR HERE.
THEY TOOK TWO SPECIES OF THE
MOSQUITOES IN THE LAB AND THEN,
AGAIN, THE SAME DAY A AT THE SAE
TIME, THEY TOOK BLOOD FROM AN
INFECTED PATIENT AND YOU HAVE TO
DO IT WITH PATIENTS WITH VIRUS
BECAUSE -- SO THEY TOOK BLOOD
FROM THE INFECTED PATIENT AND
GAVE TO THE TWO MOSQUITOES AT
THE SAME TIME AND THEY ASKED HOW
WELL ARE THEY INFECTED.
AND THEY FOUND SOMETHING
INTERESTING.
THOSE PAIR SITES WITH THE RED
GENOTYPE THAT THEY CALL C1, THEY
INFECT VERY WELL AND -- THEY
DON'T INFECT VERY WELL, SO IT'S
THE OPPOSITE FOR THOSE IN THE
HIGHLANDS.
AGAIN THE ONE IN THE HIGHLANDS
AFFECT THE MOSQUITO PRESSING IN
THE HIGH LATES -- THE COASTAL
AREA.
SO THEY KEPT IT FOR MANY YEARS,
THIS WAS PUBLISHED MANY YEARS
AGO, SO NOW WE WENT BABBL BACK D
SEQUENCED PFS47 FROM THESE
SAMPLES AND ASKED IS THERE ANY
AMINO ACID THAT CORRELATES WITH
THE ABILITY OF THE PARASITE TO
INFECT ONE MOSQUITO OR THE
OTHER?
AND WE TOOK THE MOST CLEARLY
DIFFERENT INFECTIONS THAT WERE
DIFFERENT BETWEEN THE TWO
VECTORS, SO BASICALLY WE
CONSIDER THOSE PARASITE -- THOSE
INFECTIONS WHERE THERE WAS A
SIGNIFICANT -- IN THE PREVALENCE
OF INFECTION BETWEEN THE TWO
MOSQUITOES -- SPECIES.
SO WHEN WE DID THAT, WE FOUND
SOMETHING VERY STRIKING.
IF YOU HAVE A POSITIVELY
CHARGED -- IN POSITION 27 THEN
YOUR VECTOR OF REFERENCE DOSH DO
REALLY WELL THERE BUT -- SO IF
YOU HAVE A NEGATIVELY CHARGED
RESIDUE, IT'S THE OPPOSITE.
IT'S A VERY GOOD VECTOR FOR YOU
BUT NOT MUCH LUCK IF YOU TRY TO
INFECTOINFECT ALBIMANUS.
IN RED ARE THE AMINO ACIDS IN
THE D2 DOMAIN, THIS IS IN THE
END TERMINUS, THEY'RE ALL IN THE
SAME PHASE OF PV47, THE SAME
THING IS FOUND IN PFS47, SO WE
THINK THIS MIGHT BE THE PHASE
WHERE P47 INTERACTS WITH THE
MOSQUITO RECEPTOR.
AND AGAIN, WE'RE HOPING -- WE'RE
LOOKING FORWARD TO HAVING THIS
STRUCTURE SO WE CAN REALLY MODEL
THESE INTERACTIONS AND PREDICT
COMPATIBILITY.
SO IN SHORT, WE HAVE SHOWN THE
MOSQUITO HAS AN IMMUNE SYSTEM
AND IT CAN BE EFFECTIVE IN
ELIMINATING PLASMODIUM
PARASITES.
IT'S A COMPLEX CYST THEAM
INVOLVES EPITHELIAL NITRATION,
ACTIVATION OF TEP1.
IF ANYTHING FAILS THEN THE
PARASITE CAN ESCAPE.
WE HOPE THAT THIS NEW
UNDERSTANDING WILL HELP US TO
DEVELOP NEW WAYS OF DISRUPTING
THE -- TRANSMISSION AND
PREVENTING DISEASE.
I WOULD LIKE TO FINISH BY
INTRODUCING THE PEOPLE THAT DID
THE WORK I PRESENTED TODAY.
GISELE DEMONSTRATED THAT
EPITHELIAL NITRATION WAS
ESSENTIAL FOR EFFECTIVE
ACTIVATION, HER STUDIES CLEARLY
DEMONSTRATED THAT.
LINDSAY GARVER SHOWED --
SIGNALING IS CLEARLY A KEY
PLAYER AND IS CREATING
ACTIVATION OF CASPASE.
URVASHI RAMPHUL DID A SCREEN AND
DEMONSTRATED IF YOU'RE --
SILENCING CASPASE HAS NO EFFECT
IN THE WILD TYPE PARASITE.
FOR A LONG TIME WE KNEW -- TO
ACTIVATE TEP1, HE FOUND THE
MISSING LINK IS HEMOCYTE, AND
THE RELEASE OF HEMOCYTES AND
MICROVESICLES, THAT'S AN
ESSENTIAL STEP.
IF YOU DISRUPT THE STEP, THEN
STEP ONE ACTIVATION DOESN'T
HAPPEN.
ANA PEATRIZ DISCOVERED -- SHE
IDENTIFIED THE ENZYMES THAT MAKE
THE PROSTAGLANDIN AND THEN SHOW
THAT WHEN HEMOCYTES TOUCH ON
A -- SURFACE, THEY RELEASE MICRO
VESICLES AND ARE ATTRACTED IF
YOU -- -- IS A MAJOR PLAYER,
WE'VE BEEN WORKING TOGETHER FOR
21 YEARS NOW.
HE MAPPED THE PFS47 GENE, IT WAS
A VERY CHALLENGING PROJECT THAT
TOOK ALMOST SEVEN YEARS AND A
LOT OF EFFORT, AND SOME POINT WE
WERE THINKING WE WOULD RETIRE
BEFORE WE FOUND WHAT THE GENE
WAS, BUT WE LOCKED OUT THAT
SOMEONE HAD A KNOCKOUT AND --
THAT BECAME A MAJOR CANDIDATE,
AND IT TURNED OUT TO BE THE
RIGHT THING, SO THE OTHER LAB
THAT -- SAVED US 12 YEARS OF OUR
LIVES HAVING TO DO A KNOCKOUT
FOR 40 GENES.
ALSO I WANT HE IDENTIFIED THE R,
HE DID SOME OTHER ONES LOOKING
AT THE ROLE OF EARLY TRANSITION
BETWEEN -- AND HUMANS THAT I DID
NOT PRESENT FOR THE SAKE OF
TIME.
GASPAR CANEPA WAS OUR PARASITE
ENGINEER, DETHE KNOCKOUTS AND
REPLACEMENTS, FIRST REPLACING
INDIVIDUAL AMINO ACIDS AND --
FROM DIFFERENT CONTINENTS AND
NOT SHOW IT TODAY BUT ALSO THE
HUMAN -- 47, THE PARASITE THAT
INFECTS CHIMPANZEES OR
GUERILLAS.
THEN ALSO HEE AND ALVARO WORKED
TOGETHER IN MAPPING THE REGION
WHERE ANTIBODY BINDING BLOCKED
TRANSMISSION.
AND THIAGO SILVA DID THE CELL
BIOLOGY, REALLY SHOWING WHAT ARE
THE DIFFERENCES BETWEEN WILD
TYPE AND KNOCKOUT PARASITE, AND
HE -- DOWNSTREAM FACTOR THAT IS
KEY -- BESIDES THEM, THERE WERE
MANY POSTDOCS AND STUDENTS OVER
THE YEARS THAT WERE INVOLVED IN
THESE PROJECTS, ALSO
COLLABORATORS WITHIN NMVR -- AND
RTB.
ALSO I MENTIONED FOR THE
KNOCKOUT THE GROUP IN THE
NETHERLANDS THAT GAVE US THE 54
KNOCKOUT PARASITE, AND THEN MANY
OTHER GROUPS THAT HAVE
COLLABORATED IN THE FIELD
STUDIES PROVIDING SEQUENCES FOR
THIS IN-DEPTH ANALYSIS AT A
POPULATION LEVEL.
AND FINALLY, I LOVE THIS SLIDE.
THIS IS THE PEOPLE THAT HAVE
TRAINED IN MY LAB OVER THE
YEARS, STUDENTS, POSTBAC
STUDENTS, GRADUATE STUDENTS, AND
POSTDOCS.
AND THEIR ENERGY, TALENT AND
DEDICATION IS WHAT REJUVENATES
AND ENERGIZES OUR LAB
CONSTANTLY.
ALSO THE TIME I SPEND WITH THEM
EVERY DAY IS MY FAVORITE TIME OF
THE DAY.
ALSO BEING THEIR MENTOR HAS BEEN
ONE OF THE MOST REWARDING
EXPERIENCES OF WORKING AS A
SCIENTIST.
AND ALSO I THINK THAT AS
SCIENTISTS, WE BECOME PART OF
THE LEGACY OF INDIVIDUALS WITH
STRONG CRITICAL THINKING,
BECAUSE WE UNDERSTAND THE
IMPORTANCE OF SEEKING AND
PROMOTING TRUTH AND
TRANSPARENCY.
AND I DO THINK THAT THIS IS
SOMETHING THAT IS NOW MORE
CRITICAL THAN EVER, BECAUSE WE
ARE CONSTANTLY NAVIGATING A SEA
OF INFORMATION AND
MISINFORMATION, AND PEOPLE
SOMETIMES HAVE A HARD TIME, WE
ALL HAVE A HARD TIME FINDING OUT
WHAT IS TRUE, AND WE NEED TO
DISTINGUISH WHAT IS TRUE AND
WHAT IS FAKE.
I THINK CRITICAL THINK SOMETHING
A TRAINING THAT IS GOING TO BE
VERY VALUABLE.
FINALLY, THIS WORK WAS DONE AT
NBR.
THANK YOU.
>> THANK YOU, DR. BARILLAS.
IT'S SO WELL PUT AND I REALLY
THANK YOU FOR A WONDERFUL
SEMINAR, JUST INVESTIGATIVE
RESEARCH IN EVERY POSSIBLE WAY,
CELL BIOLOGY, GEOGRAPHIC,
GENOMICS, HATS OFF TO YOU.
SO THE FLOOR IS OPEN TO
QUESTIONS.
I WILL START BY -- I'M JUST
CURIOUS, WHAT IS THE FUNCTION
FOR THIS PFS47 PROTEIN IN THE
PARASITE ITSELF?
LIKE --
>> THAT'S AN EXCELLENT QUESTION.
SO WHAT IS EMERGING IS THAT
PFS47 IS IMPORTANT TO HAVE
OPTIMAL FERTILIZATION.
IF YOU HAVE -- FOR INSTANCE, A
KNOCKOUT PARASITE, EVEN IF IT'S
INEFFICIENT AND THERE'S NO
COMPETITION, THEY STILL CAN
REPRO DAWES AND EVERYTHING IS
FINE.
BUT IF YOU HAVE A WILD TYPE AND
A KNOCKOUT PARASITE, THE WILD
TYPE WILL FERTILIZE MUCH BETTER.
>> I SEE.
>> DOES THAT MAKE SENSE?
SO WE DON'T SEE IT IN THE LAB
BECAUSE THEY'RE NOT COMPETING
WITH ANYBODY.
BUT PFS47, SO IT'S NOT AN
ABSOLUTE REQUIREMENT, BUT WHAT
IS EMERGING IS THAT IT
FACILITATES OR ENHANCES
FERTILIZATION.
>> SO THEN THE OTHER SIDE OF THE
STORY IS THE RECEPTOR.
WHAT DOES THE RECEPTOR DO
NORMALLY IN THE INSECT?
>> THAT'S A MILLION DOLLAR
QUELL.
WE DON'T KNOW WHAT IT DOES.
WE KNOW IT ESSENTIAL BECAUSE --
WELL, SO SOMETHING I DID NOT
MENTION, THAT WE HAD A
COMPLETELY DIFFERENT PROJECT
LOOKING AT WHAT IS THE RECEPTOR
THAT THE VIRUS INTERACTS WITH IN
VARIOUS MOSQUITOES AND WE DID
THE WHOLE PROJECT.
WE CHARACTERIZE THE PROTEIN --
DIFFERENT PROJECT, NOTHING TO DO
ONE WITH THE OTHER.
SO -- AND ALSO WE SHOWED THAT
EACH PROTEIN IS VERY IMPORTANT,
SO THE PICTURE THAT IS EMERGING
IS THAT YOU MAY HAVE -- TO
PATHOGENS ARE EXPLOITING THIS
MECHANISM THAT ALLOWINGS ALLOWO
SHUT DOWN CASPASES BECAUSE YOU
WANT TO INVADE THE CELL, IP FECT
THE CELL, YOU DON'T WANT THE
CELL TO DIE FROM A VIRUS WHILE
YOU'RE MULTIPLYING OR FOR A
MOSQUITO, YOU DON'T WANT TO ASK
FOR HELP AND CALL FOR THE IMMUNE
SYSTEM TO KILL YOU.
NOW WHAT IS THE FUNG FUNCTION OS
RECEPTOR?
WE DON'T KNOW, BUT WHEN WE TRIED
TO DO WITH THE KNOCKOUT,, WE
COULD NEVER GET HOMOZYGOUS
KNOCKOUTS.
IT'S ESSENTIAL.
SO IT PLAYED SOME ESSENTIAL ROLE
AND I THINK EVOLUTION OF THE
RECEPTOR HAS NOTHING TO DO WITH
THE MOSQUITOES.
BASICALLY THESE MOSQUITOES WERE
AROUND BEFORE TH THERE WERE HUM,
THEY WERE BITING DINOSAURS
ALREADY.
BUT WE CAME MUCH LATER, RIGHT?
SO THEY EMERGED IN SOUTH AMERICA
WHERE THE COLONIES WERE STILL
TOGETHER, AND WHERE DINOSAURS
WERE THEIR MAIN MEAL.
THEN EFFICIENTLY, WE -- FROM A
BARILLAS TRANSFER TODRED TO A HN
ABOUT 50,000 YEARS AGO AND THEN
MALARIA STARTED.
SO MOSQUITOES KILL ALMOST ALL
THE PARASITES, BUT ONCE IN A
WHILE, THERE'S A LUCKY PARASITE
THAT HAPPENS A PROTEIN WITH A
DIFFERENT FUNCTION THAT ALLOWS
TO TURN ON THE MOSQUITO IMMUNE
SYSTEM, AND WHEN THAT
COINCIDENCE HAPPENS, THIS MAKES
A BIG WINNER PARASITE, BECAUSE
IT NOW CAN JUMP FROM FLY FROM
PERSON TO PERSON, AND ONCE IT'S
INFECTED, ONE MOSQUITO CAN
INFECT MANY PEOPLE.
>> AND THAT BECOMES BAD NEWS FOR
US.
>> GOOD NEWS FOR THE MOSQUITO,
GOOD NEWS FOR THE PARASITE, VERY
BAD NEWS FOR US.
>> EXACTLY.
SO I WAS JUST IMPRESSED BY THE
WAY THESE RECEPTOR CLUSTERED,
AND SO IS IT A TRANSMEMBRANE
PROTEIN, AND IF THAT CLUSTERING
SOMEHOW IS REQUIRED FOR ITS
FUNCTION, IT'S JUST A LOT OF --
>> INITIALLY WE THOUGHT WE WILL
HAVE LIKE A MEMBRANE RECEPTOR
BUT IT DOESN'T SEEM TO BE THE
CASE, BUT WHEN THE PARASITE
INVADES, IT REALLY HAS TO MAKE A
PORE IN THE CELL AND THEN IT
BREAKS SITE TOE SKELETON.
IT DOESN'T FORM LIKE A VESICLE.
IT'S A VERY AGGRESSIVE INVASION.
SO ONE INTERPRETATION IS THAT
THIS MAY SEAL THE WOUND SO THAT
IMMEDIATELY, WHEN THE CELL --
BECAUSE THE PARASITE FORMS A
MEMBRANE ATTACK PROTEIN COMPLEX
AND AT SOME POINT IT START
BREAKING THE CYTOSKELETON.
AND IT TAKES THE PARASITE ABOUT
A MINUTE, A MINUTE AND A HALF
TO -- WE'VE DONE LIVE VIDEO, WE
CAN VIDEO MOSQUITOES INFEBTED
WITH A PARASITE AND WE CAN VIDEO
THE PARASITE INVADING THE RED
BLOOD CELL.
SO IT TAKES LIKE 60 TO
90 SECONDS FOR THE PARASITE TO
CROSS.
SO THAT'S VERY FAST FOR THE
CELL, THE CELL IS SLOW TO REACT,
BUT THEN YOU CAN SEND THE
SIGNALS AND CALL FOR HELP.
THE PARASITE HAS CROSSED THE
CELL, BUT THEN THE HEMOSITES
WILL COME AND THE COMPLEMENT
SYSTEM WILL COME AND THEY TAKE
CARE OF THE PARASITE.
IT CALLS FOR HELP, RIGHT?
ASKING FOR HELP.
>> I SEE.
ANY OTHER QUESTIONS?
>> SO THERE WAS APPRECIATION
FROM NIKKI, MY APOLOGIES IF I
MISPRONOUNCED THE NAME, THANK
YOU FOR AN INSPIRATIONAL TALK.
AND I COMPLETELY AGREE,
COMPLETELY INSPIRATIONAL.
CAN I ASK ONE QUESTION SINCE I
STARTED TALKING?
I FULLY APPRECIATE THE DESIRE TO
HAVE A COSTRUCTURE OF THE PFS47
PROTEIN AND ITS RECEPTOR.
AND HOW FRUSTRATING IT MUST BE
TO WANT TO KNOW WHAT THAT
STRUCTURE LOOKS LIKE.
GIVEN THAT YOU HAVE THESE GROUPS
OF THE AFRICAN VERSIONS, THE
BRAZILIAN VERSIONS OF BOTH THE
RECEPTOR AND THE 47 PROTEIN,
HAVE YOU DONE ANY MODELING OR
SORT OF PREDICTIONS, EVEN ALPHA
FOLD PREDICTIONS TO TRY AND GET
AT WHAT THE INTERACTION IS,
BECAUSE YOU HAVE SOME
DETERMINANTS OF SPECIFICITY
ALREADY.
>> YES, SO WE'VE BEEN HAVING BIG
DISCUSSION, WELL FIRST, WE ARE
NOT, YOU KNOW, MY PH.D. IS IN
BIOCHEMISTRY AND WE'VE BEEN
PLAYING AROUND WITH ALPHA FOLD.
SO WE'RE NOT SURE THAT THE
STRUCTURES THEY'RE GIVING US ARE
100%.
AND HONESTLY, WE'VE BEEN
DISCUSSING WITH ALVERO, IF YOU
KNOW SOMEONE WHO CAN HELP US,
BECAUSE WHAT WE NEED TO PUT IS
TO -- WHEN YOU TRY TO MAKE THE
COMPLEX USING ALPHA FOLD, WE
DON'T KNOW HOW TO PROVIDE THE
INFORMATION WE HAVE.
BASICALLY WE WOULD LIKE TO TELL,
THIS IS THE FACE OF THE PROTEIN
THAT SHOULD BE BINDING, BECAUSE
THAT'S WHERE ALL THE IMPORTANT
POLYMORPHISMS ARE, AND IF WE
HAVE THE MODEL WHERE THE
DIFFERENCES IN THE RECEPTORS --
HAVE MANY DIFFERENCES.
SO THE RECEPTORS ARE LIKE MUCH
MORE -- LIKE 80% IDENTICAL,
THEY'RE NOT AS SIMILAR AS PFS47.
SO FOR THE RECEPTOR, WE DON'T
KNOW HOW TO PREDICT HOW IT WILL
BIND.
SO PFS47, IT'S PRETTY CLEAR BUT
THE RECEPTOR IS ABOUT
80-SOMETHING% SO IT HAS LAKE 20%
DIFFERENCE IN AMINO -- IT NOT SO
OBVIOUS HOW THEY FIT TOGETHER,
PARTICULARLY THE ONES FROM NEW
WORLD AND AFRICA BECAUSE THEY
ARE SO DISTANT, BUT IF YOU
REMEMBER, THEY'RE ALSO THE ONES
THAT HAVE THE MOST EXTREME
PHENOTYPES.
SO THE JUMP -- SO MANY PEOPLE
HAVE ASIA NO THE NEW WORLD, NONE
OF THESE PARASITES ARE ABLE TO
ESTABLISH A TRANSMISSION CYCLE
IN THE NEW WORLD.
SO THIS IS A VERY STRONG
NEGATIVE SELECTION.
BUT WE HOPE TO GET AT LEAST ONE
PAIR TO WORK BECAUSE ONCE I
THINK WE GET ONE PAIR WORKING
THEN WE CAN MODEL ALL THE REST.
THAT'S OUR HOPE.
BUT WE'RE VERY CLOSE.
WE'RE NOW BEGINNING -- WE HAVE
ALMOST ENOUGH, WE'RE OPTIMIZING
THE PREPS TO REALLY STABILIZE
THE COMPLEX.
ALSO THE RECEPTOR IS
CHALLENGING, BECAUSE WE THINK IT
NORMALLY INTERACTS WITH OTHER
PROTEINS, SO IT DOESN'T HAVE TO
BE ALONE IN SOLUTION, IT FALLS
OUT OF SOLUTION IF YOU LACK AT
IT SIDEWAYS.
SO WE HAVE TO STABILIZE IT, BIND
THE COMPLEX AND WE HAVE TO --
IT'S ONLY STABLE UNDER VERY HIGH
SALT, SO WE HAVE TO FIND -- ONCE
IT'S BOUND TO SOMETHING, IT'S
STABLE.
BUT THE STABILITY OF THE
RECEPTOR HAS BEEN A BIG
CHALLENGE.
>> WELL, THANK YOU.
SO YOU KNOW, DR. BARILLAS, YOU
ARE SOMEONE WHO HAS TRAVELED FAR
BY DISTANCE, CAREER,
SCIENTIFICALLY IN SPEAKING.
SO FOR THE AUDIENCE, THERE'S A
QUESTION FROM THE FELLOWS THAT
WHAT WOULD BE THE ONE ADVICE YOU
WOULD GIVE TO THE NEW GENERATION
OF WOMEN OR ANY SCIENTIST FOR
THAT MATTER, BUT ESPECIALLY
MINORITY SCIENTISTS?
>> WELL, I THINK IN GENERAL, I
WOULD SAY YOU NEED TO LOVE WHAT
YOU DO, THAT'S ONE THING.
ENJOY WHAT YOU'RE DOING.
PEOPLE TELL ME OH, YOU WENT TO
SCAL LIKE 20-SOMETHING YEARS.
YEAH, BUT I ENJOY EVERY DAY OF
IT, YOU KNOW?
I'M A NERD.
SO I ENJOY BEING A NERD, RIGHT?
AS LONG AS YOU ENJOY THE PATH,
NO?
THE SECOND ONE IS DON'T BE
AFRAID TO TRY THINGS.
WHEN I STARTED MY PH.D., I
JOINED A LAB AND IT TURNED OUT
TO BE A DISASTER AND AFTER TWO
YEARS, I HAD TO DROP THE PH.D.
THESIS AND START AGAIN.
BUT THEN, YOU KNOW, THREE YEARS
LATER, 2 1/2 YEARS LATER, LESS
THAN THREE YEARS LATER, I
GRADUATED.
I DID A WHOLE NEW PROJECT
STARTING FROM SCRATCH.
BUT I HAD LEARNED SO MUCH IN
THOSE TWO YEARS THAT I HAD
MATURED SCIENTIFICALLY.
SO SOMETIMES THERE'S SETBACKS
BUT I KNEW I WAS IN THE WRONG
PLACE AND HAD TO CHANGE.
SO DON'T BE AFRAID, EVEN IF THE
COST IS HIGH, IF YOU SEE THERE'S
SOMETHING THAT IS REALLY NOT
GOING -- DON'T BE AFRAID.
DON'T BE AFRAID TO ASK.
WHEN I DID MY -- I HAVE AN
ANECDOTE, WHEN I WAS DOING MY
PH.D. IN ARIZONA, THEY HAD A
MEETING WITH THE PEOPLE FROM THE
MA CAR TUR FOUNMCARTHUR FOUNDATD
TO SWITCH TO MOSQUITOES SO THERE
WERE VERY FEW PEOPLE IN THE
MOLECULAR TOOLS, WE CLONED THE
FIRST EVER IN MOSQUITO, SO THEY
INVITED ALL THE MAIN SCIENTIST,
AND MY PH.D. ADVISOR, HE WAS
SUPERVISOR GENEROUS, TOM
WELLAMS, HE TOLD ME FRIDAY,
DON'T SCHEDULE ANYTHING, I
VOLUNTEER, YOU'RE GOING TO BE
CHANGING -- THEY HAD CAROUSELS
WITH SLIDES, HE SAID NO, NO,
YOU'RE GOING TO THANK ME BECAUSE
THE BEST PEOPLE WORKING WITH
VECTORS AND THE FUTURE OF WHERE
THIS FIELD IS GOING IS GOING TO
BE -- IT'S A CLOSED MEETING, YOU
ARE GOING TO BE THE ONLY STUDENT
THAT WILL GET IN BECAUSE YOU'RE
GOING TO BE CHANGING SLIDES.
SO YOU LISTEN TO ALL THE TALKS
AND THEN -- SO I WENT THERE, AND
I LISTENED TO THE TALK OF --
FROM HARVARD.
HE BLEW MY MIND AWAY AND I WAS
SO AFRAID OF APPROACHING HIM.
BECAUSE I WAS LIKE -- TALKING TO
PROFESSOR -- FROM HARVARD.
I THOUGHT I WAS TOTALLY OUT OF
MY LEAGUE.
BUT THEN I REMEMBERED, I HAD A
PROFESSOR THAT TOLD ME, AS A
STUDENT YOU HAVE ONE BIG THING
GOING FOR YOU.
YOU HAVE NOTHING, AND THEN YOU
HAVE NOTHING TO LOSE.
SO DON'T FORGET THAT, RIGHT?
SO WHAT IS THE WORST THING THAT
CAN HAPPEN?
YOU STAY EXACTLY WHERE YOU ARE,
SO YOU HAVE NOTHING TO LOSE AND
MUCH TO GAIN.
SO I WENT AND TALKED TO HIM,
LONG STORY SHORT AFTER HE
INVITED ME TO GIVE A TALK AND WE
DISCUSSED, I ENDED UP DOING MY
POSTDOC WITH HIM AT HARVARD AND
THEN AT EMBL, AND I THINK IN
TERMS OF TRAINING, THAT PUT ME
IN A VERY GOOD POSITION WITH TOP
LEVEL TRAINING, RIGHT?
WE WERE THE GENERATION THAT
BUILT THE TOOLS, AND THAT WAS
ALSO FRUSTRATING AS POSTDOCS
BECAUSE THEY HAD ALL THE TOOLS
AND WE HAVE NOTHING, WE HAVE TO
BUILD EVERYTHING FROM SCRATCH.
BUT I THINK THAT NOW WE ARE 20
YEARS, 25 YEARS LATER, WE'RE AT
A POINT WHERE WE HAVE ENOUGH
TOOLS TO ASK GOOD QUESTIONS,
RIGHT?
AND DO INTERESTING WORK.
>> THAT'S GREAT.
THE STORY ALSO TELLS US TO TRY
TO ISAIAS THE OPPORTUNITY WHEN
IT AVAILABLE.
AND YES, I THINK THAT'S REALLY A
GREAT BOTTOM LINE.
ENJOY, AND NOT BE AFRAID OF
FAILURES.
>> AND CHANGE, RIGHT?
>> AND CHANGE.
RIGHT.
>> AND ALSO I HAVE BEEN VERY
LUCKY BECAUSE EVERYWHERE W WHERE
I'VE BEEN, THIS IS AN ACTIVITY
TO PROMOTE WOMEN SCIENTIST, BUT
I'VE HAD AMAZING MALE
SCIENTISTS, COLLEAGUES, MENTORS
THAT HAVE BEEN JUST SUPER
HELPFUL.
SO I THINK THAT YOU KNOW, IF YOU
LOOK CAREFULLY, THEY'RE ALWAYS
IN EACH DEPARTMENT, YOU WILL
FIND, YOU KNOW, COLLEAGUES THAT
ARE SUPPORTIVE, THAT ARE OPEN,
MIKE MENTORS, I GIVE YOU THIS
EXAMPLE OF GIVING THIS
OPPORTUNITY TO BE IN THE FIELD,
THAT WAS MIKE WELLS, AND SECOND
ONE WAS FOR -- BOTH OF THEM
PASSED AWAY SOME YEARS AGO, BUT
THEY LEFT -- THEY WERE SUCH
EXCELLENT MENTORS THAT YOU FEEL
THAT IT IS YOUR DUTY TO FOLLOW
THEIR STEPS AND CONTRIBUTE FROM
THE NEXT GENERATION, BECAUSE
THEY LEFT A LEGACY WITH YOU.
>> THAT'S WONDERFUL.
SO I DON'T SEE ANY OTHER
QUESTIONS IN THE CHAT.
SO IF THERE'S NO OTHER QUESTIONS
THAT I CAN SEE, I'D LIKE TO
CLOSE THE SEMINAR AND THANK YOU
AGAIN, DR. BARILLAS, FOR YOUR
WONDERFUL PRESENTATION.
>> THANK YOU VERY MUCH.
>> YES, THANK YOU.
EXCELLENT.
>> BYE.