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GREETINGS AND SALUTATIONS!

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I'M DAN KASTNER, THE SCIENTIFIC

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DIRECTOR EMERITUS OF THE

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NATIONAL HUMAN GENOME RESEARCH

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INSTITUTE.

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AND STILL ACTIVE PHYSICIAN,

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SCIENTIST, IN THE INTRAMURAL

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RESEARCH PROGRAM OF THE NIH.

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IT IS MY ENORMOUS PLEASURE TO

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WELCOME YOU TO THIS, THE 16th

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INSTALLMENT IN THE 2025 SEASON

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OF DEMYSTIFYING MEDICINE.

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AN EXCITING COURSE THAT BRIDGES

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DEVELOPMENT IN BIOLOGY,

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ENGINEERING, COMPUTER SCIENCE,

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WITH MEDICINE.

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YOU CAN SEE IN THE UPPER

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RIGHT-HAND CORNER OF THE SLIDE

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THE ICONIC IMAGE OF THE

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CONSTRUCTION OF THE BROOKLYN

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BRIDGE, BACK IN THE LATE

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1800s, UP ON THAT CATWALK YOU

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CAN SEE A PHYSICIAN, PROBABLY

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FROM MANHATTAN, AND A BASIC

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SCIENTIST FROM BROOKLYN, HAVING

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A DISCUSSION UP ON THE CATWALK

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TALKING ABOUT THE LATEST

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DEVELOPMENTS IN PHAGOCYTOSIS AND

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THE ACTIONS OF ANTIBODIES.

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AND AS IT WOULD HAPPEN, TODAY

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WE'RE GOING TO BE TALKING ABOUT

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A RELATED TOPIC, NEUTROPHIL

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NETs, AND JUST BEFORE WE GET

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INTO THAT LET ME REMIND YOU THAT

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THIS SERIES IS ONE THAT OCCURS

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TUESDAY AFTERNOONS FROM 3 TO 5

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IN THE AFTERNOON JANUARY THROUGH

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MAY.

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YOU CAN WATCH ON VIDEOCAST AT

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THE LINK SHOWN ON THE SCREEN,

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FOR THOSE OF YOU WHO WANT

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CONTINUING MEDICAL EDUCATION

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CREDIT THE CODE IS 58795.

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AGAIN, THAT'S 58795.

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DURING THE TWO PRESENTATIONS,

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AND WE'LL HAVE TWO PRESENTATIONS

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AND THEN A LIVELY DISCUSSION

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SESSION FOR THE LAST HALF HOUR

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OR SO, BUT IF YOU HAVE QUESTIONS

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THAT OCCUR TO YOU DURING THE

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PRESENTATIONS, PLEASE USE  THE

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"SEND LIVE FEEDBACK" BUTTON ON

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THE VIDEOCAST DISPLAY.

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ALL THE PREVIOUS SESSIONS ARE

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ARCHIVED AT THE LINK SHOWN ON

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THE SCREEN AND FOR ADDITIONAL

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INFORMATION YOU CAN USE THAT

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DEMYSTIFYING MEDICINE E-MAIL

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ADDRESS.

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OF COURSE, HERE WE ARE BRIDGING

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SILOS AT NIH SINCE 2003.

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MOVING ON TO OUR FIRST SPEAKER,

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WHO WILL BE TALKING TO US ABOUT

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NEUTROPHIL EXTRACELLULAR TRAPS,

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NETs, WE HAVE THE GOOD DR.

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ARTURO ZYCHLINSKY, WHO IS THE

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DIRECTOR OF THE DEPARTMENT FOR

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CELLULAR MICROBIOLOGY AT THE MAX

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PLANCK INSTITUTE IN BERLIN. 

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OBTAINED A BACHELOR'S DEGREE, I

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DON'T SPEAK SPANISH, IN MEXICO

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CITY.

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AND THEN HE RECEIVED HIS Ph.D.

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IN IMMUNOLOGY AT THE ROCKEFELLER

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UNIVERSITY.

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HE DID AN EMBO  POSTDOCTORAL

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FELLOW AND MOVED TO SKIRBALL

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BEFORE ASSUMING HIS CURRENT

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POSITION AT DIRECTOR AT THE MAX

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PLANCK INSTITUTE.

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HE'S RECIPIENT OF A NUMBER OF

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PRESTIGIOUS AWARDS, INCLUDING

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THE IRMA T HIRSCHL AWARD, THE

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GROHE AWARD, A MEMBER OF THE

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GERMAN NATIONAL ACADEMY OF

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SCIENCES LEOPOLDINA AND EUROPEAN

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ACADEMY OF MICROBIOLOGY, WORKING

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ON THE ROLE OF NEUTROPHIL

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EXTRACELLULAR TRAPS IN HEALTH

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AND DISEASE, ON THE

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ANTIMICROBIAL CAPACITY OF

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HISTONES AND EVOLUTION AND THEIR

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FUNCTION IN GENE REGULATION AND

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DURING MULTI-CELLULAR

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DEVELOPMENT, AND HE'S ALSO BEEN

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INTERESTED IN THE INFLAMMASOME,

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AND ITS ACTIVATION.

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AND HOW THE IMMUNE SYSTEM

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MEASURES THE DANGER AND ADAPTS

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ITS RESPONSE.

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OUR SECOND SPEAKER IS DR.

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MARIANA KAPLAN, FROM HERE AT THE

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NIH, WHO IS GOING TO BE TALKING

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TO US ABOUT FRIENDLY FIRE, HOW

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NETs CONTRIBUTE TO

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INFLAMMATION AND AUTOIMMUNITY.

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SHE'S AN NIH DISTINGUISHED

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INVESTIGATOR, DEPUTY SCIENTIFIC

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DIRECTOR, AND CHIEF OF THE

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SYSTEMIC AUTOIMMUNITY BRANCH AT

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THE NATIONAL INSTITUTE FOR

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ARTHRITIS, MUSCULOSKELETAL AND

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SKIN DISEASES.

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SHE'S ALSO AN ADJUNCT PROFESSOR

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OF INTERNAL MEDICINE, UNIVERSITY

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OF MICHIGAN AND ADJUNCT

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PROFESSOR IN THE GRADUATE

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PROGRAM IN IMMUNOLOGY AT THE

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UNIVERSITY OF PENNSYLVANIA. 

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SHE OBTAINED HER MEDICAL DEGREE

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FROM THE NATIONAL AUTONOMOUS

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UNIVERSITY OF MEXICO, GOT HER

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INTERNAL MEDICINE TRAINING AT

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THE NATIONAL INSTITUTE OF

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MEDICAL SCIENCES AND NUTRITION

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IN MEXICO CITY, RHEUMATOLOGY

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TRAINING AT THE UNIVERSITY OF

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MICHIGAN.

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SHE TOO IS THE RECIPIENT OF A

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NUMBER OF PRESTIGIOUS AWARDS

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INCLUDING THE HENRY KUNKEL YOUNG

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INVESTIGATOR AWARD, EDMUND

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DUBOISE MEMORIAL LECTURESHIP,

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BASIC AND TRANSLATIONAL

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INVESTIGATOR AWARD AND EVELYN

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HESS AWARD FROM LUPUS FOUNDATION

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OF AMERICA.

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SHE'S ALSO A MEMBER OF THE

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AMERICAN SOCIETY FOR CLINICAL

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INVESTIGATION, ASSOCIATION OF

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AMERICAN PHYSICIANS, AND

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NATIONAL ACADEMY OF MEDICINE.

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HER RESEARCH GROUP WORKS ON HOW

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THE INNATE IMMUNE SYSTEM

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PROMOTES AUTOIMMUNITY AND END OR

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G

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AN DAMAGE IN NETs, SYSTEMIC

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AUTOIMMUNE DISEASE, NOVEL

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THERAPEUTIC TARGETS TO PREVENT

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PREMATURE VASCULAR DAMAGE, AND

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SHE IS INVOLVED IN CLINICAL

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TRIALS TO REDUCE BLOOD VESSEL

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DYSFUNCTION AND MITIGATE ORGAN

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DAMAGE.

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WELL, THAT'S BEEN A MOUTHFUL,

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BUT NOW IT'S TIME TO TURN THINGS

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OVER TO OUR SPEAKERS, AND WE'LL

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START WITH THE GOOD DR.

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ZYCHLINSKY TALKING ABOUT

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NEUTROPHIL EXTRACELLULAR TRAPS.

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ARTURO, PLEASE, TAKE IT AWAY.

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>> THANK YOU, DAN, FOR
SUCH A

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NICE INTRODUCTION.

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IT'S A PLEASURE TO TALK HERE,

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ESPECIALLY AS A COMPANION OF

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MARIANA.

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I WANTED TO DOUBLECHECK YOU CAN

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SEE MY SCREEN.

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>> INDEED.

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>> OKAY, GOOD.

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SO, I'M GOING TO TALK ABOUT

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NEUTROPHILS, THE MOST ABUNDANT

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WHITE BLOOD CELLS IN HUMAN

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BLOOD.

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THEY MAKE AROUND 70% OF ALL THE

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WHITE BLOOD CELLS AND THEY ARE

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ESSENTIAL FOR IMMUNITY.

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PEOPLE OR KIDS THAT ARE BORN

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WITH DEFECT IN NEUTROPHIL

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BIOLOGY OR PEOPLE BECAUSE OF

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CHEMOTHERAPY HAVE A LOW NUMBER

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AND ARE SUSCEPTIBLE TO

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INFECTION, ALSO NEUTROPHILS ARE

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THE FIRST RESPONDERS AFTER

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INFECTION.

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SO IN THIS SLIDE I TRIED TO

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ILLUSTRATE HOW A MICROBE STARTS

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AN INFECTION BY BREAKING

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EPITHELIAL BARRIER, RIGHT UNDER

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THE EPITHELIUM THERE ARE CELLS

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LIKE DENDRITIC CELLS AND MICRO

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PHAGES SIGNAL THERE'S AN

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INFECTION, PRO-INFLAMMATORY

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CYTOKINES AND  AND EXTRAVASITESA

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HUNDRED-FOLD MORE EFFECTIVE AT

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KILLING PHAGOCYTES LIKE

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MACROPHAGES, SET UP THE STAGE

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FOR THE RECRUITMENT OF MORE

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NEUTROPHILS IF INFECTION IS NOT

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CONTROLLED.

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THE NEUTROPHIL IS EASY TO

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RECOGNIZE IN BLOOD, IT HAS A

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NUCLEUS THAT'S LOBULATED AS

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OPPOSED TO MOST CELLS WITH

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CIRCLE OR ELLIPSOID NUCLEUS.

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NOW I'M SHOWING AN IMAGE OF A

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NEUTROPHIL ISOLATED FROM HUMAN

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BLOOD BY DENSITY CENTRIFUGATION.

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ONE OF THE JOBS OF THE

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NEUTROPHIL IS EAT BACTERIA, THIS

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IS A CULTURE OF NEUTROPHILS,

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INFECTED WITH SHIGELLA, YOU CAN

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SEE HOW NEUTROPHIL SEES THE

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BACTERIA, ENGULFS IT AND

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GRANULES THAT CONTAIN

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ANTIMICROBIALS, THIS IS

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PHAGOCYTOSIS DESCRIBED OVER A

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HUNDRED YEARS AGO, ONE OF THE

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VERY EFFECTIVE WAYS HOW

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NEUTROPHILS DISPOSE OF BACTERIA.

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MANY YEARS AGO WE DESCRIBED A

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SECOND WAY OF HOW NEUTROPHILS

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CAN ACTIVATE AND KILL MICROBES,

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BY FORMATION OF NETs.

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NETs STANDS FOR NEUTROPHIL

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EXTRACELLULAR TRAPS, A CULTURE

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OF HUMAN NEUTROPHILS ISOLATED

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AND ACTIVATED IN VITRO AND

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STAINED BY IMMUNOFLUORESCENCE

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WITH MARKERS.

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I'M SHOWING PROJECTIONS.

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YOU CAN SEE THAT WE'LL START

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WITH ELASTASE, A PROTEASE THAT'S

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EXPRESSED IN THE GRANULE OF

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NEUTROPHILS, YOU CAN SEE THESE

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NEUTROPHILS, NOT ACTIVATED, IT'S

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IN THE GRANULES AND ALSO IN THE

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NETs YOU CAN SEE HERE.

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HISTONES ARE THE PROTEINS THAT

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COVER CHROMATIN, COVER DNA TO

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FORM CHROMATIN, AND YOU CAN SEE

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NON-ACTIVATED NEUTROPHIL ONLY

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THE NUCLEUS IS ACTIVATED AND YOU

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CAN SEE HERE HOW CLEARLY

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LOBULATED THE NUCLEI OF

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NEUTROPHILS ARE, AND ALSO USE

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DNA STAINS AND HERE YOU CAN SEE

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THE LOBULATED NUCLEUS OF THE

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INTACT NEUTROPHILS BUT YOU CAN

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SEE CHROMATIN THAT IS DNA ALSO

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PRESENT IN NETs.

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NETS ARE COMPOSED OF CHROMATIN

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WITH ADDITION OF SPECIFIC

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PROTEINS THAT COME MOSTLY FROM

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NEUTROPHILS.

256
00:11:18,359 --> 00:11:20,828
ONE OF THE QUESTIONS THAT HAS

257
00:11:20,828 --> 00:11:22,663
INTERESTED MY LAB FOR THE LAST

258
00:11:22,663 --> 00:11:27,268
15 YEARS IS HOW THESE NETs ARE

259
00:11:27,268 --> 00:11:27,635
MADE.

260
00:11:27,635 --> 00:11:30,371
BECAUSE NETs, WE THINK THAT

261
00:11:30,371 --> 00:11:34,508
CELL DEATH OF A CELL IS REQUIRED

262
00:11:34,508 --> 00:11:35,443
FOR THE EXTRAVASATION OF

263
00:11:35,443 --> 00:11:37,211
CHROMATIN SO WE WANTED TO

264
00:11:37,211 --> 00:11:39,213
UNDERSTAND HOW THE CHROMATIN IS

265
00:11:39,213 --> 00:11:44,251
EXPELLED FROM THE CELL INTACT.

266
00:11:44,251 --> 00:11:47,722
SO, THROUGHOUT MY TALK I'LL USE

267
00:11:47,722 --> 00:11:52,059
MOSTLY EXPERIMENTS DONE IN MY

268
00:11:52,059 --> 00:11:53,461
LAB BY ISOLATING NEUTROPHILS AND

269
00:11:53,461 --> 00:11:56,597
ONE OF THE FIRST THINGS WE FOUND

270
00:11:56,597 --> 00:11:58,199
OUT IS THAT THE CELL ACTUALLY

271
00:11:58,199 --> 00:12:01,836
HAS TO DIE IN ORDER TO RELEASE

272
00:12:01,836 --> 00:12:03,104
ITS CHROMATIN CONTENT.

273
00:12:03,104 --> 00:12:04,071
THIS IS ANOTHER MOVIE.

274
00:12:04,071 --> 00:12:06,474
THIS IS, AGAIN, A CULTURE OF

275
00:12:06,474 --> 00:12:06,874
HUMAN NEUTROPHILS.

276
00:12:06,874 --> 00:12:10,044
YOU CAN SEE THE LOBULATED

277
00:12:10,044 --> 00:12:11,512
NUCLEUS WITH DNA DYE THAT IS

278
00:12:11,512 --> 00:12:15,216
HERE IN PINK.

279
00:12:15,216 --> 00:12:20,254
AND IT CONTAINS TWO DNA DYEs,

280
00:12:20,254 --> 00:12:21,956
ONE PERMITTABLE IN PINK, YOU CAN

281
00:12:21,956 --> 00:12:25,459
SEE THE NUCLEUS ALREADY PINK.

282
00:12:25,459 --> 00:12:36,003
THE SECOND IS DNA IMPERMEABLE IN

283
00:12:37,905 --> 00:12:38,339
GREEN.

284
00:12:38,339 --> 00:12:41,142
YOU CAN SEE HOW THE NUCLEUS GOES

285
00:12:41,142 --> 00:12:43,477
THROUGH A SPECIFIC PROCESS IT

286
00:12:43,477 --> 00:12:45,946
GETS SMALLER, IT EXPANDS AND

287
00:12:45,946 --> 00:12:47,348
FINALLY ALL CHROMATIN IS

288
00:12:47,348 --> 00:12:49,050
RELEASED FOR NETs AGAIN, I'LL

289
00:12:49,050 --> 00:12:51,285
PLAY THIS MOVIE AGAIN SO YOU CAN

290
00:12:51,285 --> 00:12:54,989
SEE, FOR EXAMPLE, HERE HOW THE

291
00:12:54,989 --> 00:12:58,325
NUCLEUS CONTRACTS, EXPANDS,

292
00:12:58,325 --> 00:12:58,793
FORMS NETs.

293
00:12:58,793 --> 00:13:00,795
SO, THIS IS SIMILAR EXPERIMENT

294
00:13:00,795 --> 00:13:02,396
AS THE ONE I SHOWED, A DIFFERENT

295
00:13:02,396 --> 00:13:04,532
WAY OF LOOKING AT THE

296
00:13:04,532 --> 00:13:04,832
NEUTROPHILS.

297
00:13:04,832 --> 00:13:10,471
IN THIS CASE USING TRANSITION

298
00:13:10,471 --> 00:13:11,439
ELECTROMICROSCOPY, A NEUTROPHIL

299
00:13:11,439 --> 00:13:12,640
HAS NOT BEEN ACTIVATED.

300
00:13:12,640 --> 00:13:15,609
YOU CAN SEE TWO OF THE MANY FOUR

301
00:13:15,609 --> 00:13:16,310
OR FIVE LOBULATED NEUTROPHILS

302
00:13:16,310 --> 00:13:19,080
HAVE AND YOU CAN ALSO SEE THE

303
00:13:19,080 --> 00:13:22,983
DIFFERENT GRANULES THAT COME IN

304
00:13:22,983 --> 00:13:23,451
DIFFERENT FLAVORS.

305
00:13:23,451 --> 00:13:25,052
NOW, ONE HOUR AFTER ACTIVATION

306
00:13:25,052 --> 00:13:34,128
ALL THE LOBULATION HAS BEEN

307
00:13:34,128 --> 00:13:35,963
LOST, THERE'S VERY SPECIFIC

308
00:13:35,963 --> 00:13:37,064
ALTERATION ALSO STARTING TO

309
00:13:37,064 --> 00:13:38,933
HAPPEN, FOR EXAMPLE, TO THE

310
00:13:38,933 --> 00:13:41,702
NUCLEAR MEMBRANE. 

311
00:13:41,702 --> 00:13:42,503
TWO HOURS AFTER ACTIVATION, NOW

312
00:13:42,503 --> 00:13:45,840
WE CAN SEE THIS IS THE

313
00:13:45,840 --> 00:13:48,275
CHROMATIN, THE REMNANT, NOW THE

314
00:13:48,275 --> 00:13:51,512
CHROMATIN IS IN DIRECT CONTACT

315
00:13:51,512 --> 00:13:53,714
WITH THE CYTOPLASM OF THE CELL.

316
00:13:53,714 --> 00:13:55,082
YOU CAN SEE VESICLES, I'M TRYING

317
00:13:55,082 --> 00:14:02,723
TO POINT OUT WITH MY CURSOR, AND

318
00:14:02,723 --> 00:14:04,825
THESE ARE THE REMNANTS.

319
00:14:04,825 --> 00:14:08,229
WHEN WE DO MICROSCOPE WE WITH

320
00:14:08,229 --> 00:14:09,530
MARKERS WE CAN SEE THIS.

321
00:14:09,530 --> 00:14:11,832
THIS IS THE MOMENT WHERE THE

322
00:14:11,832 --> 00:14:15,436
CHROMATIN GETS IN TOUCH WITH

323
00:14:15,436 --> 00:14:16,804
CYTOPLASM, GRANULES ARE

324
00:14:16,804 --> 00:14:19,039
DISSOLVED, WE BELIEVE THIS IS A

325
00:14:19,039 --> 00:14:20,608
TIME WHERE SPECIFIC PROTEINS ARE

326
00:14:20,608 --> 00:14:24,078
ATTACHED TO THE CHROMATIN IN

327
00:14:24,078 --> 00:14:26,147
ORDER TO FORM THE NETs.

328
00:14:26,147 --> 00:14:28,349
THIS IS IMAGED THREE HOURS AFTER

329
00:14:28,349 --> 00:14:31,452
ACTIVATION, THESE ARE THE

330
00:14:31,452 --> 00:14:32,219
PRE-NETs, MODIFIED CHROMATINS

331
00:14:32,219 --> 00:14:34,288
WE BELIEVE THE CELL WILL EXPLODE

332
00:14:34,288 --> 00:14:38,559
VERY SOON TO RELEASE THE NETs

333
00:14:38,559 --> 00:14:41,228
OUT OF THE CYTOPLASMIC MEMBRANE.

334
00:14:41,228 --> 00:14:43,063
THIS IS A SCHEME I TRIED TO

335
00:14:43,063 --> 00:14:47,434
AGAINST AGAIN SHOWING HOW THE

336
00:14:47,434 --> 00:14:49,470
NUCLEUS FIRST IS COMPACTED, THEN

337
00:14:49,470 --> 00:14:51,438
EXPANDED, AND HOW THE DIFFERENT

338
00:14:51,438 --> 00:14:53,307
GRANULAR PROTEINS GET INTO THE

339
00:14:53,307 --> 00:14:55,609
NUCLEUS TO EVENTUALLY MAKE

340
00:14:55,609 --> 00:14:57,311
NETs.

341
00:14:57,311 --> 00:15:00,414
SO, VERY EARLY ON, WE REALIZE

342
00:15:00,414 --> 00:15:02,616
THAT REACTIVE OXYGEN SPECIES

343
00:15:02,616 --> 00:15:05,319
WERE RELEVANT FOR NET FORMATION.

344
00:15:05,319 --> 00:15:07,855
REACTIVE OXYGEN SPECIES ARE

345
00:15:07,855 --> 00:15:08,889
PRODUCED BY SPECIFIC ENZYMES,

346
00:15:08,889 --> 00:15:14,395
WHERE MOW  MOLECULAR OXYGEN IS

347
00:15:14,395 --> 00:15:16,397
CONVERTED, NEUTROPHIL IS VERY

348
00:15:16,397 --> 00:15:16,997
EFFECTIVE, THE MOST EFFECTIVE

349
00:15:16,997 --> 00:15:22,970
CELL IN THE BODY TO PRODUCE

350
00:15:22,970 --> 00:15:23,237
PEROXIDE.

351
00:15:23,237 --> 00:15:25,072
PEROXIDE MUTATES INTO HYDROGEN

352
00:15:25,072 --> 00:15:28,576
PEROXIDE, THIS IS A REACTION

353
00:15:28,576 --> 00:15:31,011
THAT HAPPENS NATURALLY BUT THERE

354
00:15:31,011 --> 00:15:32,813
ARE TWO ENZYMES TO PROCESS THESE

355
00:15:32,813 --> 00:15:38,185
MUTATIONS THAT ALLOWS THIS

356
00:15:38,185 --> 00:15:38,752
REACTION.

357
00:15:38,752 --> 00:15:40,254
FINALLY MELA PEROXIDASE CONVERTS

358
00:15:40,254 --> 00:15:49,096
HYDROGEN PEROXIDE INTO 

359
00:15:49,096 --> 00:15:56,870
HYPOCHLORIC ACID, YOU ARE

360
00:15:56,870 --> 00:15:57,805
FAMILIAR, USED TO DISINFECT, AND

361
00:15:57,805 --> 00:16:02,509
ONE IS BLEACH.

362
00:16:02,509 --> 00:16:05,279
THEY IMMEDIATELY OXIDIZE

363
00:16:05,279 --> 00:16:07,414
PROTEINS, LIPIDS, NUCLEIC ACIDS,

364
00:16:07,414 --> 00:16:09,783
EVERYTHING BY THIS VERY ACTIVE

365
00:16:09,783 --> 00:16:10,117
OXIDANT.

366
00:16:10,117 --> 00:16:13,721
WE THINK THESE MOLECULES ACT AS

367
00:16:13,721 --> 00:16:14,321
PROTEIN MODIFIERS THAT ALLOW

368
00:16:14,321 --> 00:16:16,690
THIS FUNCTION OF NETs.

369
00:16:16,690 --> 00:16:24,398
COMING BACK TO NET FORMATION

370
00:16:24,398 --> 00:16:29,336
NEUTROPHILS WITHIN HALF AN HOUR,

371
00:16:29,336 --> 00:16:34,041
USING INHIBITORS NADPH OXIDASE

372
00:16:34,041 --> 00:16:35,242
WAS REQUIRED.

373
00:16:35,242 --> 00:16:37,111
IT'S INTERESTING TO CONFIRM

374
00:16:37,111 --> 00:16:42,783
GENETICALLY, AND WE STARTED TO

375
00:16:42,783 --> 00:16:49,156
WORK WITH PATIENTS WITH CHRONIC

376
00:16:49,156 --> 00:16:49,390
DISEASE.

377
00:16:49,390 --> 00:16:54,028
MUTATIONS IN ADPH OXIDASE CANNOT

378
00:16:54,028 --> 00:16:54,461
PRODUCE SUPEROXIDE.

379
00:16:54,461 --> 00:16:56,930
WHEN WE TAKE NEUTROPHILS FROM

380
00:16:56,930 --> 00:16:58,899
THESE PATIENTS, YOU CAN SEE

381
00:16:58,899 --> 00:17:01,502
NEUTROPHILS LOOK PERFECTLY WELL.

382
00:17:01,502 --> 00:17:06,373
AND THIS IS CONVENTIONAL

383
00:17:06,373 --> 00:17:10,911
MICROSCOPY, GRANULAR MARK

384
00:17:10,911 --> 00:17:12,913
NUCLEAR ELASTASE, WHEN WE

385
00:17:12,913 --> 00:17:15,482
ACTIVATE THESE CELLS THE CELLS

386
00:17:15,482 --> 00:17:16,984
LOOK MUCH FLATTER, LIKE IN THE

387
00:17:16,984 --> 00:17:20,821
MOVIE I SHOWED AT THE START OF

388
00:17:20,821 --> 00:17:23,490
THE TALK OF THE NEUTROPHIL

389
00:17:23,490 --> 00:17:26,126
INFECTED WITH BACTERIA BUT THE

390
00:17:26,126 --> 00:17:28,762
NUCLEUS ARE COMPLETELY LOBULATED

391
00:17:28,762 --> 00:17:36,704
AND THESE CELLS CANNOT MAKE

392
00:17:36,704 --> 00:17:39,139
NETs.

393
00:17:39,139 --> 00:17:41,475
AND IN ORDER TO COMPLEMENT THE

394
00:17:41,475 --> 00:17:43,210
MUTATION IN THESE PATIENTS, WE

395
00:17:43,210 --> 00:17:45,112
DID IT GENETICALLY AS I'LL SHOW

396
00:17:45,112 --> 00:17:46,313
IN A SMALL ANECDOTE TOWARDS THE

397
00:17:46,313 --> 00:17:48,682
END OF THE TALK.

398
00:17:48,682 --> 00:17:55,789
WE HAVE ADD  HYDROGEN PEROXIDE

399
00:17:55,789 --> 00:18:00,394
AND SHOW NOW WE CAN MAKE THE

400
00:18:00,394 --> 00:18:01,695
CGD, NEUTROPHILS MAKE NETs

401
00:18:01,695 --> 00:18:05,566
BECAUSE WE'RE GOING AFTER THE

402
00:18:05,566 --> 00:18:08,635
NADPH OXIDASE, COMPLEMENTING THE

403
00:18:08,635 --> 00:18:08,902
PHENOTYPE.

404
00:18:08,902 --> 00:18:11,238
WE PROPOSE BASED ON THIS GENETIC

405
00:18:11,238 --> 00:18:14,375
DATA AND PHARMACOLOGICAL DATA

406
00:18:14,375 --> 00:18:17,578
THAT THE NADPH OXIDASE WAS

407
00:18:17,578 --> 00:18:20,948
REQUIRED FOR NET FORMATION, THE

408
00:18:20,948 --> 00:18:24,585
STIMULI WE USED FOR NET

409
00:18:24,585 --> 00:18:26,019
FORMATION ARE MAINLY MICROBES,

410
00:18:26,019 --> 00:18:29,156
EITHER BACTERIA OR FUNGI, AND

411
00:18:29,156 --> 00:18:31,158
THIS IS PATHWAY I'M DESCRIBING

412
00:18:31,158 --> 00:18:33,427
ALSO TRUE FOR, FOR EXAMPLE, FOR

413
00:18:33,427 --> 00:18:35,629
NITROGEN, THERE ARE OTHER FORMS

414
00:18:35,629 --> 00:18:37,798
OF NET FORMATION THAT USE

415
00:18:37,798 --> 00:18:40,434
TENTATIVE PATHWAY NOT WELL

416
00:18:40,434 --> 00:18:42,870
DESCRIBED THAT DOESN'T REQUIRE

417
00:18:42,870 --> 00:18:43,771
THE FORMATION.

418
00:18:43,771 --> 00:18:46,573
BACK TO THE REACTIVE OXYGEN

419
00:18:46,573 --> 00:18:48,509
SPECIES, WE ASSUME THAT THESE

420
00:18:48,509 --> 00:18:51,445
NADPH OXIDASE WAS REQUIRED, ALSO

421
00:18:51,445 --> 00:18:55,149
WOULD NEED THE PEROXIDASE

422
00:18:55,149 --> 00:18:59,186
MUTATIONS, DEFECTS IN MILA

423
00:18:59,186 --> 00:19:00,721
PEROXIDASE FUNCTION ARE COMMON,

424
00:19:00,721 --> 00:19:07,127
QUITE OFTEN, BUT THESE ARE ONLY

425
00:19:07,127 --> 00:19:09,296
HYPERMARKS.

426
00:19:09,296 --> 00:19:12,065
MPO-DEFICIENT PATIENTS ARE RARE,

427
00:19:12,065 --> 00:19:13,167
PHENOTYPE REMINISCENT OF CGD

428
00:19:13,167 --> 00:19:15,436
PATIENT SUSCEPTIBLE TO

429
00:19:15,436 --> 00:19:15,769
INFECTION.

430
00:19:15,769 --> 00:19:20,374
WE MANAGE TO COLLECT A FEW

431
00:19:20,374 --> 00:19:22,242
MPO-DEFICIENT PATIENTS AND COULD

432
00:19:22,242 --> 00:19:24,144
SHOW THEY ALSO CANNOT MAKE

433
00:19:24,144 --> 00:19:24,344
NETs.

434
00:19:24,344 --> 00:19:27,448
THIS IS A CONTROL OF A HEALTHY

435
00:19:27,448 --> 00:19:28,282
DONOR.

436
00:19:28,282 --> 00:19:30,350
I HOPE YOU ALREADY RECOGNIZE THE

437
00:19:30,350 --> 00:19:32,085
STRANDS ARE THE NETs THAT ARE

438
00:19:32,085 --> 00:19:34,021
STAYING BOTH WITH GRANULAR

439
00:19:34,021 --> 00:19:38,325
MARKER LIKE NEUTROPHIL ELASTIC

440
00:19:38,325 --> 00:19:42,029
AND CHROMATIN MARK, THIS

441
00:19:42,029 --> 00:19:42,629
ANTIBODY, BUT MPO DEFICIENT

442
00:19:42,629 --> 00:19:45,766
PATIENTS THE NEUTROPHILS ARE

443
00:19:45,766 --> 00:19:47,100
UNABLE TO MAKE NETs.

444
00:19:47,100 --> 00:19:49,036
THE NEUTROPHIL ELASTIC IS STILL

445
00:19:49,036 --> 00:19:53,040
IN THE GRANULES, NUCLEI ARE

446
00:19:53,040 --> 00:19:55,209
STILL LOBULATED.

447
00:19:55,209 --> 00:20:05,752
WE THING THE FIRST STEP REQUIRES

448
00:20:07,221 --> 00:20:10,290
FORMATION. -- FORMATION OF ROS.

449
00:20:10,290 --> 00:20:12,459
WE TAKE NEUTROPHILS ISOLATED

450
00:20:12,459 --> 00:20:14,761
FROM HEALTHY HUMANS, WE CAN SEE

451
00:20:14,761 --> 00:20:15,796
BEFORE ACTIVATION THE NEUTROPHIL

452
00:20:15,796 --> 00:20:17,498
ELASTIC IS IN THE GRANULES,

453
00:20:17,498 --> 00:20:19,366
AFTER 90 MINUTES OF ACTIVATION

454
00:20:19,366 --> 00:20:21,602
WE CAN SEE THAT ELASTIC IS

455
00:20:21,602 --> 00:20:23,437
STARTING TO MIGRATE INTO THE

456
00:20:23,437 --> 00:20:25,172
NUKE 

457
00:20:25,172 --> 00:20:25,405
NUCLEUS.

458
00:20:25,405 --> 00:20:26,974
WE DON'T UNDERSTAND HOW THE

459
00:20:26,974 --> 00:20:28,041
NEUTROPHIL ELASTIC GETS OUT OF

460
00:20:28,041 --> 00:20:30,878
THE GRANULES OR HOW IT GETS INTO

461
00:20:30,878 --> 00:20:33,280
THE NUCLEUS, IT'S A SMALL

462
00:20:33,280 --> 00:20:33,881
PROTEIN SO COULD PASS THROUGH

463
00:20:33,881 --> 00:20:36,550
THE NUCLEAR PORE BUT THIS IS AN

464
00:20:36,550 --> 00:20:38,852
AREA OF INVESTIGATION THAT WE

465
00:20:38,852 --> 00:20:43,156
REALLY HAVEN'T PURSUED.

466
00:20:43,156 --> 00:20:49,496
SO, I'M SHOWING THAT THE

467
00:20:49,496 --> 00:20:49,930
NEUTROPHIL ELASTASE

468
00:20:49,930 --> 00:20:54,167
TRANSLOCATES, AND WE TURNED TO

469
00:20:54,167 --> 00:20:56,403
MICE TO TELL IF IT WAS REQUIRED

470
00:20:56,403 --> 00:20:57,371
FOR NET FORMATION.

471
00:20:57,371 --> 00:20:59,439
I'M ONLY GOING TO SHOW ONE PIECE

472
00:20:59,439 --> 00:21:00,007
OF DATA.

473
00:21:00,007 --> 00:21:03,477
AND THIS IS A SCANNING ELECTRON

474
00:21:03,477 --> 00:21:07,314
MICROGRAPH OF THE LUNG OF MICE

475
00:21:07,314 --> 00:21:09,383
INFECTED WITH KLEBSIELLA, QUITE

476
00:21:09,383 --> 00:21:11,051
A DANGEROUS BACTERIA THAT CAUSES

477
00:21:11,051 --> 00:21:15,455
PNEUMONIA AND WE'RE LOOKING AT

478
00:21:15,455 --> 00:21:20,027
THE HIGH MAGNIFICATION OF THE

479
00:21:20,027 --> 00:21:20,260
ALVEOLI.

480
00:21:20,260 --> 00:21:21,862
YOU CAN SEE BACTERIA ARE COVERED

481
00:21:21,862 --> 00:21:23,530
IN FIBROUS MATERIAL WHICH ARE

482
00:21:23,530 --> 00:21:24,831
THE NETs.

483
00:21:24,831 --> 00:21:26,833
WHEN WE INFECT NEUTROPHIL

484
00:21:26,833 --> 00:21:29,536
ELASTIC KNOCKOUT MICE YOU CAN

485
00:21:29,536 --> 00:21:31,071
SEE THE BACTERIA, AND WHAT YOU

486
00:21:31,071 --> 00:21:34,575
CANNOT SEE IN THE NET, BUT IN

487
00:21:34,575 --> 00:21:36,610
PARALLEL EXPERIMENT I'M NOT

488
00:21:36,610 --> 00:21:37,678
SHOWING INFILTRATION OF

489
00:21:37,678 --> 00:21:39,446
NEUTROPHILS INTO THE LUNG OF THE

490
00:21:39,446 --> 00:21:47,187
NEUTROPHIL ELASTICS  ELASTICS MT

491
00:21:47,187 --> 00:21:48,388
EFFECTIVE, THEY ARE THERE BUT

492
00:21:48,388 --> 00:21:50,023
UNABLE TO MAKE NETs.

493
00:21:50,023 --> 00:21:55,228
SECOND STEP AFTER FORMATION OF

494
00:21:55,228 --> 00:21:57,064
ROS TRANSLOCATION FROM GRANULES

495
00:21:57,064 --> 00:21:59,399
INTO NUCLEUS ALLOWING THE

496
00:21:59,399 --> 00:22:00,434
FORMATION OF NETs.

497
00:22:00,434 --> 00:22:03,704
SO, I SHOWED YOU THIS IMAGE

498
00:22:03,704 --> 00:22:04,137
BEFORE.

499
00:22:04,137 --> 00:22:05,706
AND I MENTIONED HERE THE

500
00:22:05,706 --> 00:22:08,742
CHROMATIN IS IN DIRECT CONTACT

501
00:22:08,742 --> 00:22:10,377
WITH CYTOPLASM AND NUCLEAR

502
00:22:10,377 --> 00:22:15,215
MEMBRANE HAS BEEN COMPLETELY

503
00:22:15,215 --> 00:22:16,817
VACULATED AND DISAPPEARED.

504
00:22:16,817 --> 00:22:18,919
ANOTHER EXAMPLE IN BIOLOGY, THIS

505
00:22:18,919 --> 00:22:21,989
IS MITOSIS THROUGH CLASSICAL

506
00:22:21,989 --> 00:22:23,557
CELL DIVISIONS, I SHOULD MENTION

507
00:22:23,557 --> 00:22:27,427
NEUTROPHILS ARE TERMINALLY

508
00:22:27,427 --> 00:22:30,197
DIFFERENTIATED CELLS THAT WILL

509
00:22:30,197 --> 00:22:31,198
NOT DIVIDE, SHORT HALF-LIFE

510
00:22:31,198 --> 00:22:35,969
MEASURED IN HOURS, NOT DAYS.

511
00:22:35,969 --> 00:22:38,572
SO, A POSTDOC IN MY LAB PROPOSED

512
00:22:38,572 --> 00:22:40,507
THE IDEA THAT THERE WERE COMMON

513
00:22:40,507 --> 00:22:50,384
MECHANISMS IN THE FORMATION OF

514
00:22:50,384 --> 00:22:51,685
MITOSIS AND NETOSIS.

515
00:22:51,685 --> 00:22:55,756
DURING THE CELL CYCLE AFTER

516
00:22:55,756 --> 00:22:59,459
MITOSIS THERE'S A PHASE HISTONES

517
00:22:59,459 --> 00:23:02,229
AND DNA SYNTHESIZE, GOES INTO A

518
00:23:02,229 --> 00:23:03,964
PHASE AND CELLS DIVIDE.

519
00:23:03,964 --> 00:23:09,336
NEUTROPHILS ARE HERE IN THE G

520
00:23:09,336 --> 00:23:09,970
SERA, TERMINALLY DIFFERENTIATED,

521
00:23:09,970 --> 00:23:11,004
ESCAPE CELL CYCLE.

522
00:23:11,004 --> 00:23:12,739
SO, ONE OF THE FIRST EXPERIMENTS

523
00:23:12,739 --> 00:23:21,648
THAT WE DID WAS TO SHOW THAT

524
00:23:21,648 --> 00:23:26,319
ACTUALLY MITOGENS CAN INDUCE,

525
00:23:26,319 --> 00:23:28,822
THIS IS AN IMAGE, IF WE ACTIVATE

526
00:23:28,822 --> 00:23:30,624
THE NEUTROPHILS CAN MAKE NETs

527
00:23:30,624 --> 00:23:30,991
EFFECTIVELY.

528
00:23:30,991 --> 00:23:33,260
THIS IS THE FIRST COMMONALITY

529
00:23:33,260 --> 00:23:35,462
BETWEEN NET FORMATION AND

530
00:23:35,462 --> 00:23:37,964
MITOSIS, BOTH CAN BE ACTIVATED

531
00:23:37,964 --> 00:23:40,200
BY MITOGENS.

532
00:23:40,200 --> 00:23:43,070
THERE'S A MARKER KI-67 FOR

533
00:23:43,070 --> 00:23:47,641
PROLIFERATION, NOT EXPRESSED BY

534
00:23:47,641 --> 00:23:50,777
NEUTROPHILS, THEY ARE NOT

535
00:23:50,777 --> 00:23:51,244
DIVIDING.

536
00:23:51,244 --> 00:23:53,680
WE TAKE NAIVE NEUTROPHILS, THE

537
00:23:53,680 --> 00:23:56,817
DNA STAIN, THEY DON'T EXPRESS

538
00:23:56,817 --> 00:23:57,184
Ki67.

539
00:23:57,184 --> 00:23:58,685
WE SEE THE CLEAR ACTIVATION WHEN

540
00:23:58,685 --> 00:24:01,321
WE ACTIVATE, THIS IS THE SECOND

541
00:24:01,321 --> 00:24:06,760
CLUE THAT THE NEUTROPHILS ARE

542
00:24:06,760 --> 00:24:08,195
USING THE MITOTIC PATHWAY IN

543
00:24:08,195 --> 00:24:13,033
ORDER TO MAKE NETs. 

544
00:24:13,033 --> 00:24:14,968
ACTUALLY, THE Ki67 POSITIVITY

545
00:24:14,968 --> 00:24:19,239
HAPPENS ALSO IN VIVO, THIS IS A

546
00:24:19,239 --> 00:24:23,443
SECTION OF PATIENT THAT DIED OF

547
00:24:23,443 --> 00:24:27,180
FUNGAL ABSCESS IN THE BRAIN,

548
00:24:27,180 --> 00:24:29,116
GOOD AT RECRUITING NEUTROPHILS,

549
00:24:29,116 --> 00:24:34,821
HERE WE'RE LABELING IN RED, MANY

550
00:24:34,821 --> 00:24:38,225
IN THE AREA, AND WE'RE USING

551
00:24:38,225 --> 00:24:39,593
ALSO Ki67 TO IDENTIFY

552
00:24:39,593 --> 00:24:40,827
NEUTROPHILS, SO THERE ARE

553
00:24:40,827 --> 00:24:42,028
NEUTROPHILS LIKE THIS ONE OR

554
00:24:42,028 --> 00:24:45,632
THIS ONE, YOU CAN SEE THE

555
00:24:45,632 --> 00:24:48,835
LOBULATED NEUTROPHIL AND NOT

556
00:24:48,835 --> 00:24:54,207
ACTIVATED YET BUT ARE

557
00:24:54,207 --> 00:24:55,342
NEUTROPHILS Ki67 POSITIVE, WE

558
00:24:55,342 --> 00:25:01,081
BELIEVE ABOUT TO MAKE NETs IN

559
00:25:01,081 --> 00:25:01,548
VIVO.

560
00:25:01,548 --> 00:25:07,254
MITOSIS REQUIRES ACTIVATION OF

561
00:25:07,254 --> 00:25:09,156
CYCLIN DEPENDENT KINASE AND THIS

562
00:25:09,156 --> 00:25:12,559
IS AN EXPERIMENT WITH MICE, NOT

563
00:25:12,559 --> 00:25:14,628
HUMAN CELLS.

564
00:25:14,628 --> 00:25:17,130
IF WE TAKE NEUTROPHILS FROM

565
00:25:17,130 --> 00:25:18,365
WILDTYPE OR KNOCKOUT MICE THEY

566
00:25:18,365 --> 00:25:23,436
DON'T MAKE NETs BUT IF WE

567
00:25:23,436 --> 00:25:25,672
ACTIVATE EITHER WITH MITOGEN OR

568
00:25:25,672 --> 00:25:29,476
MICROBE WITH CANDIDA ALBICANS

569
00:25:29,476 --> 00:25:30,811
THE NEUTROPHIL FROM WILD TYPE

570
00:25:30,811 --> 00:25:34,948
MAKE NETS, SEVERELY HAMPERED IN

571
00:25:34,948 --> 00:25:37,918
ANIMALS CDK 6 IS KNOCKED OUT.

572
00:25:37,918 --> 00:25:41,888
THE FINAL EXAMPLE I SHOWED HOW

573
00:25:41,888 --> 00:25:50,697
NET FORMATION REQUIRES MITOSIS

574
00:25:50,697 --> 00:25:52,032
WE CAN SEE DIFFERENT TUBULES

575
00:25:52,032 --> 00:25:58,205
MADE IN CELL BUT IN ACTIVATED

576
00:25:58,205 --> 00:25:58,905
CELLS MAKE CENTROSOME, GOING TO

577
00:25:58,905 --> 00:26:02,909
THE TWO SIDES OF THE NUCLEUS OF

578
00:26:02,909 --> 00:26:03,777
THE CELL.

579
00:26:03,777 --> 00:26:06,479
WE'VE NEVER SEEN MITOTIC

580
00:26:06,479 --> 00:26:06,746
STIMULUS.

581
00:26:06,746 --> 00:26:17,290
SOMEHOW THIS IS THE FORMATION OF

582
00:26:22,162 --> 00:26:22,996
CENTROSOMES, WE'VE NOT FIGURED

583
00:26:22,996 --> 00:26:23,430
OUT.

584
00:26:23,430 --> 00:26:25,565
THIS IS A SUMMARY OF THIS PART.

585
00:26:25,565 --> 00:26:28,468
WHEN WE ACTIVATE NEUTROPHILS TO

586
00:26:28,468 --> 00:26:31,438
MAKE NETs, THEY EXPRESS

587
00:26:31,438 --> 00:26:34,007
Ki67, THEY REQUIRE ACTIVITY OF

588
00:26:34,007 --> 00:26:35,342
CYCLIN DEPENDENT KINASE AND

589
00:26:35,342 --> 00:26:38,511
ESSENTIAL FOR MIGRATION BEFORE

590
00:26:38,511 --> 00:26:42,749
THE FORMATION OF NETs.

591
00:26:42,749 --> 00:26:45,719
IN ORDER TO SEE THE THE ACTUAL

592
00:26:45,719 --> 00:26:47,220
FINAL EFFECTOR OF NETs WE DID

593
00:26:47,220 --> 00:26:51,858
A LARGE SCREEN, LOOKED FOR

594
00:26:51,858 --> 00:26:55,328
INHIBITORS OF NET FORMATION BY

595
00:26:55,328 --> 00:26:57,664
ISOLATING NEUTROPHILS FROM

596
00:26:57,664 --> 00:26:59,099
HUMANS AND THEN ACTIVATE TO MAKE

597
00:26:59,099 --> 00:27:02,869
NETs WITH A  MITOGEN IN

598
00:27:02,869 --> 00:27:05,405
PRESENCE OR ABSENCE OF

599
00:27:05,405 --> 00:27:07,307
INHIBITORS, SCREEN FOR

600
00:27:07,307 --> 00:27:11,111
COMPOUNDS, CLASSICAL SCREEN,

601
00:27:11,111 --> 00:27:13,880
SCREENED IN COLLABORATION WITH

602
00:27:13,880 --> 00:27:15,248
INSTITUTE ALSO MAX PLANCK, AND A

603
00:27:15,248 --> 00:27:18,351
COMPANY THAT DEPENDS ON THEM,

604
00:27:18,351 --> 00:27:19,753
OVER 250,000 COMPOUNDS, WHICH IF

605
00:27:19,753 --> 00:27:24,291
YOU THINK THE CELLS THAT WERE

606
00:27:24,291 --> 00:27:25,992
ISOLATED FOR HUMANS AND THEY

607
00:27:25,992 --> 00:27:28,728
ONLY LIVE FOR SIX HOURS, IT WAS

608
00:27:28,728 --> 00:27:30,030
REALLY A VERY HEROIC EFFORT OF

609
00:27:30,030 --> 00:27:33,566
THE PEOPLE WHO ACTUALLY MADE

610
00:27:33,566 --> 00:27:36,236
THESE ASSAYS, I MENTIONED THEM.

611
00:27:36,236 --> 00:27:37,070
WE FOUND SEVERAL INHIBITORS,

612
00:27:37,070 --> 00:27:38,505
WORKING THROUGH THEM.

613
00:27:38,505 --> 00:27:41,675
ONE OF THE INHIBITORS WE

614
00:27:41,675 --> 00:27:48,815
IDENTIFY IS INHIBITOR OF

615
00:27:48,815 --> 00:27:52,419
GASDERMIN THAT ALLOWS RELEASE OF

616
00:27:52,419 --> 00:27:54,154
NETs, CONTROL CELL ACTIVATED

617
00:27:54,154 --> 00:27:56,923
IN ABSENCE OF INHIBITORS,

618
00:27:56,923 --> 00:28:00,860
SIMILAR TO ONE I SHOWED EARLIER.

619
00:28:00,860 --> 00:28:04,130
IN THE PRESENCE OF GASDERMIN

620
00:28:04,130 --> 00:28:06,199
INHIBITORS CELLS ARE STUCK AND

621
00:28:06,199 --> 00:28:08,401
THERE IS SOME EXPANSION OF THE

622
00:28:08,401 --> 00:28:13,640
NUCLEUS BUT VERY LITTLE NET

623
00:28:13,640 --> 00:28:14,240
FORMATION.

624
00:28:14,240 --> 00:28:17,811
SO, THIS IS WHERE WE ARE IN THE

625
00:28:17,811 --> 00:28:19,179
PROCESS OF NET FORMATION.

626
00:28:19,179 --> 00:28:21,715
AND WE SEE THAT THERE IS

627
00:28:21,715 --> 00:28:23,883
REQUIREMENT OF ROS.

628
00:28:23,883 --> 00:28:26,920
NEUTROPHIL ELASTICS MAKE IT TO

629
00:28:26,920 --> 00:28:30,123
THE NUCLEUS, ALSO REQUIRED,

630
00:28:30,123 --> 00:28:31,758
GASDERMIN-D IS ACTIVATED WHICH

631
00:28:31,758 --> 00:28:32,959
ALLOWS NET FORMATION, AND THIS

632
00:28:32,959 --> 00:28:36,896
IS HOW IT GOES.

633
00:28:36,896 --> 00:28:40,533
THERE'S A RECEPTOR ACTIVATED,

634
00:28:40,533 --> 00:28:42,369
FORMATION OF ROS, NPO,

635
00:28:42,369 --> 00:28:44,437
NEUTROPHIL ELASTIC MIGRATES TO

636
00:28:44,437 --> 00:28:46,573
THE NEUTROPHIL, CELL EXPANDS,

637
00:28:46,573 --> 00:28:50,610
ACTIVATION OF GASDERMIN, FINALLY

638
00:28:50,610 --> 00:28:54,314
NET FORMATION.

639
00:28:54,314 --> 00:28:55,348
SO, NETs HAVE THREE

640
00:28:55,348 --> 00:28:56,649
INDEPENDENT FUNCTION.

641
00:28:56,649 --> 00:28:58,785
ONE OF THEM IS ANTIMICROBIAL

642
00:28:58,785 --> 00:29:00,420
CELLS, VERY IMPORTANT IN KILLING

643
00:29:00,420 --> 00:29:01,421
MICROBES.

644
00:29:01,421 --> 00:29:02,622
SECOND ONE IS THEY ARE

645
00:29:02,622 --> 00:29:03,023
PRO-INFLAMMATORY.

646
00:29:03,023 --> 00:29:05,392
I'LL MENTION A LITTLE BUILT ON

647
00:29:05,392 --> 00:29:06,793
THIS BECAUSE THIS IS WHERE I

648
00:29:06,793 --> 00:29:08,228
THINK MARIANA IS GOING TO EXPAND

649
00:29:08,228 --> 00:29:10,830
IN THE SECOND PART OF THIS

650
00:29:10,830 --> 00:29:11,398
SEMINAR.

651
00:29:11,398 --> 00:29:15,301
AND NETs ALSO HAVE A POWERFUL

652
00:29:15,301 --> 00:29:17,370
PRO COAGULANT ACTIVITY, THE WORK

653
00:29:17,370 --> 00:29:21,541
OF DENISE WAGNER, AND I WILL NOT

654
00:29:21,541 --> 00:29:23,076
MENTION THAT FURTHER.

655
00:29:23,076 --> 00:29:25,512
SO, THE ANTIMICROBIAL ACTIVITY

656
00:29:25,512 --> 00:29:27,781
OF NETs HAS DIFFERENT EFFECTS,

657
00:29:27,781 --> 00:29:32,252
THIS IS A SCANNING EM OF

658
00:29:32,252 --> 00:29:33,620
NEUTROPHILS INFECTED WITH

659
00:29:33,620 --> 00:29:35,221
SHIGELLA, WE MENTIONED GRAM

660
00:29:35,221 --> 00:29:38,992
NEGATIVE THAT CAUSES DYSENTERY,

661
00:29:38,992 --> 00:29:39,993
OR STAPHYLOCOCCUS AURA, GRAM

662
00:29:39,993 --> 00:29:44,664
POSITIVE THAT CAUSES ALL KINDS

663
00:29:44,664 --> 00:29:50,637
OF DISEASE LIKE IMPETIGO AND OH

664
00:29:50,637 --> 00:29:55,375
OTITIS MEDIA, ET CETERA.

665
00:29:55,375 --> 00:29:57,377
THEY PRESENT THE BACTERIA

666
00:29:57,377 --> 00:30:03,249
USUALLY AN IONIC, NETS ARE

667
00:30:03,249 --> 00:30:04,150
CATIONIC, AND PREVENT

668
00:30:04,150 --> 00:30:05,885
DISSEMINATIONS SHOWN BY SEVERAL

669
00:30:05,885 --> 00:30:13,827
LABS THAT MANY PATHOGENS HAVE

670
00:30:13,827 --> 00:30:15,562
DEVELOPED THAT ALLOW THEM TO

671
00:30:15,562 --> 00:30:17,497
ESCAPE.

672
00:30:17,497 --> 00:30:24,270
THE SECOND FUNCTION IS KILL

673
00:30:24,270 --> 00:30:27,807
MICROBES, WE ACTIVATE

674
00:30:27,807 --> 00:30:29,309
NEUTROPHILS, BY FOUR HOURS NO

675
00:30:29,309 --> 00:30:30,977
NEUTROPHILS ARE LEFT ALIVE.

676
00:30:30,977 --> 00:30:32,378
WE INFECT CULTURES, WAIT 20

677
00:30:32,378 --> 00:30:35,048
MINUTES, SEE HOW MANY BACTERIA

678
00:30:35,048 --> 00:30:36,382
ARE ACTUALLY KILLED.

679
00:30:36,382 --> 00:30:37,450
WHEN WE TAKE THESE EXPERIMENTS

680
00:30:37,450 --> 00:30:39,519
AND DO IT IN THE PRESENCE OF

681
00:30:39,519 --> 00:30:42,622
NETs, WE SEE THAT IN 20

682
00:30:42,622 --> 00:30:47,427
MINUTES AROUND 25% OR  35% OF

683
00:30:47,427 --> 00:30:47,927
CULTURE OF SHIGELLA OR

684
00:30:47,927 --> 00:30:48,962
STAPHYLOCOCCUS ARE KILLED.

685
00:30:48,962 --> 00:30:50,930
IF WE DO CONTROLLED EXPERIMENT

686
00:30:50,930 --> 00:30:55,735
WHERE WE DESTROY THE NETs BY

687
00:30:55,735 --> 00:30:58,438
DIGESTING WE DON'T SEE KILLING.

688
00:30:58,438 --> 00:31:01,441
ALTERNATIVE WAY IS TO USE

689
00:31:01,441 --> 00:31:02,575
ANTI-HISTONE ANTIBODIES.

690
00:31:02,575 --> 00:31:04,144
SO HERE WE ALLOW THE NEUTROPHILS

691
00:31:04,144 --> 00:31:06,813
TO MAKE NETs.

692
00:31:06,813 --> 00:31:09,382
BEFORE INFECTING THEM WE USE

693
00:31:09,382 --> 00:31:10,350
ANTI-HISTONE ANTIBODIES.

694
00:31:10,350 --> 00:31:11,484
OR CONTROL ANTIBODY.

695
00:31:11,484 --> 00:31:14,387
IF WE USE CONTROL ANTIBODY WE

696
00:31:14,387 --> 00:31:18,791
STILL SEE KILLING WITH SOME OF

697
00:31:18,791 --> 00:31:21,194
THE EXAMPLES THAT WE USE BUT

698
00:31:21,194 --> 00:31:23,062
WHEN WE USE ANTI-HISTONE

699
00:31:23,062 --> 00:31:25,598
ANTIBODY WE COMPLETELY BLOCK THE

700
00:31:25,598 --> 00:31:27,100
ANTIMICROBIAL ACTIVITY OF

701
00:31:27,100 --> 00:31:27,433
NETs.

702
00:31:27,433 --> 00:31:32,772
I THINK THIS EXPERIMENT HAS TWO

703
00:31:32,772 --> 00:31:33,306
POSSIBLE EXPLANATIONS, NOT

704
00:31:33,306 --> 00:31:36,309
NECESSARILY MUTUALLY EXCLUSIVE.

705
00:31:36,309 --> 00:31:38,478
ONE IS THAT THE ANTI-HISTONE

706
00:31:38,478 --> 00:31:39,679
ANTIBODIES PREVENT BINDING OF

707
00:31:39,679 --> 00:31:42,749
THE BACTERIA TO NETs, BINDING

708
00:31:42,749 --> 00:31:44,617
IS REQUIRED.

709
00:31:44,617 --> 00:31:47,453
ALSO SHOWS ANTI-HISTONES ARE

710
00:31:47,453 --> 00:31:48,354
VERY IMPORTANT ANTIMICROBIALS,

711
00:31:48,354 --> 00:31:53,626
ACTUALLY A DISCOVERY FROM THE

712
00:31:53,626 --> 00:31:58,731
LATE 50s, THE ACTIVITY WAS

713
00:31:58,731 --> 00:32:01,367
FIRST DESCRIBED BY HIRSCH, AND

714
00:32:01,367 --> 00:32:04,537
THEY ARE PROBABLY THE MOST

715
00:32:04,537 --> 00:32:05,205
POTENT ANTIMICROBIALS THAT

716
00:32:05,205 --> 00:32:07,440
EUKARYOTES HAVE.

717
00:32:07,440 --> 00:32:17,917
THEY KILL MICROBES, CALCIUM

718
00:32:21,487 --> 00:32:22,288
MAGNESIUM.

719
00:32:22,288 --> 00:32:23,623
SO I THINK IT'S INTERESTING TO

720
00:32:23,623 --> 00:32:27,894
THINK WHY IN EVOLUTION HISTONES

721
00:32:27,894 --> 00:32:29,963
EVOLVING EUKARYOTES AS

722
00:32:29,963 --> 00:32:32,232
ANTIMICROBIALS AND IN ORDER TO

723
00:32:32,232 --> 00:32:33,099
PROTECT LARGE EUKARYOTES.

724
00:32:33,099 --> 00:32:34,834
SO ONE OF THE QUESTIONS THAT I

725
00:32:34,834 --> 00:32:36,402
THINK IS RELEVANT TO US WHETHER

726
00:32:36,402 --> 00:32:38,204
NETs ARE RELEVANT IN

727
00:32:38,204 --> 00:32:38,538
INFECTION.

728
00:32:38,538 --> 00:32:41,674
AND I WILL USE ONE EXAMPLE WITH

729
00:32:41,674 --> 00:32:43,443
AN  N-of-1, BUT I THINK IT'S A

730
00:32:43,443 --> 00:32:44,677
VERY DRAMATIC VERSION.

731
00:32:44,677 --> 00:32:47,080
THIS IS A SCANNING ELECTRON

732
00:32:47,080 --> 00:32:49,249
MICROSCOPE. 

733
00:32:49,249 --> 00:32:53,086
WHAT YOU SEE HERE IS

734
00:32:53,086 --> 00:32:58,858
ASPERGILLUS, HIGH C IS LONG,

735
00:32:58,858 --> 00:33:00,660
NEUTROPHIL TRYING TO

736
00:33:00,660 --> 00:33:01,561
PHAGOCYTOSE.

737
00:33:01,561 --> 00:33:04,697
THE NEUTROPHIL WILL NOT BE

738
00:33:04,697 --> 00:33:05,865
EFFECTIVE.

739
00:33:05,865 --> 00:33:07,267
SO PHAGOCYTOSIS IS NOT EFFECTIVE

740
00:33:07,267 --> 00:33:08,768
IN FUNGAL INFECTIONS.

741
00:33:08,768 --> 00:33:11,437
THIS IS THE  N-of-1.

742
00:33:11,437 --> 00:33:15,441
THIS IS A PATIENT, CGD PATIENT,

743
00:33:15,441 --> 00:33:19,012
I ALREADY MENTIONED CGD PATIENTS

744
00:33:19,012 --> 00:33:20,880
HAVE MUTATIONS IN NADPA OXIDASE.

745
00:33:20,880 --> 00:33:24,817
THIS KID WAS SUFFERING FROM A

746
00:33:24,817 --> 00:33:26,753
SEVERE ASPERGILLUS INFECTION, HE

747
00:33:26,753 --> 00:33:32,258
WAS IN A HOSPITAL IN SWITZERLAND

748
00:33:32,258 --> 00:33:34,427
AND THE PHYSICIAN AND SCIENTIST

749
00:33:34,427 --> 00:33:37,196
TAKING CARE OF THIS KID REACHED

750
00:33:37,196 --> 00:33:37,530
OUT TO US.

751
00:33:37,530 --> 00:33:40,433
THIS IS THE -- THE KID WAS VERY

752
00:33:40,433 --> 00:33:43,436
SICK, THE DECISION WAS THAT THE

753
00:33:43,436 --> 00:33:45,338
KID WOULD NOT SURVIVE BONE

754
00:33:45,338 --> 00:33:46,372
MARROW TRANSPLANTATION, ONE OF

755
00:33:46,372 --> 00:33:49,776
THE WAYS THEY ARE TREATED, AND

756
00:33:49,776 --> 00:33:54,580
INSTEAD THE TEAM DECIDED TO DO

757
00:33:54,580 --> 00:33:55,114
GENE THERAPY.

758
00:33:55,114 --> 00:33:58,518
SO, THIS IS ANALYSIS OF THE

759
00:33:58,518 --> 00:34:02,422
NADPH OXIDASE SUBUNIT WHERE THE

760
00:34:02,422 --> 00:34:06,726
KID ARE MUTATION OR ROS, AND

761
00:34:06,726 --> 00:34:11,097
CONTROLLED DONOR HAS NADPA

762
00:34:11,097 --> 00:34:18,137
OXIDASE PRODUCES ROS, AFTER GENE

763
00:34:18,137 --> 00:34:20,473
THERAPY THERE WAS A CHIMERISM,

764
00:34:20,473 --> 00:34:24,644
30% OF CELLS STARTED EXPRESSING

765
00:34:24,644 --> 00:34:28,981
SUBUNIT, AND MADE ROS.

766
00:34:28,981 --> 00:34:30,383
NOW, WE ALSO ISOLATED THE

767
00:34:30,383 --> 00:34:32,919
NEUTROPHILS FROM THIS PATIENT.

768
00:34:32,919 --> 00:34:33,920
AGAIN, THE CONTROL DISEASE ONLY

769
00:34:33,920 --> 00:34:36,089
A DNA STAIN, YOU CAN SEE THE

770
00:34:36,089 --> 00:34:38,925
CONTROL MAKES EFFECTIVE NETs.

771
00:34:38,925 --> 00:34:39,892
BEFORE GENE THERAPY THE PATIENT,

772
00:34:39,892 --> 00:34:44,664
NUKE LEE 

773
00:34:44,664 --> 00:34:46,799
NUCLEI ARE LOBULATED, 30% OF NET

774
00:34:46,799 --> 00:34:48,234
FORMATION AFTER GENE THERAPY OF

775
00:34:48,234 --> 00:34:49,335
THIS KID.

776
00:34:49,335 --> 00:34:51,571
AND WE ALSO ISOLATED THE

777
00:34:51,571 --> 00:34:54,107
ASPERGILLUS STRAIN FROM THE LUNG

778
00:34:54,107 --> 00:34:56,642
OF THIS PATIENT AND THEN WE

779
00:34:56,642 --> 00:34:58,978
TESTED WHETHER THE NEUTROPHILS

780
00:34:58,978 --> 00:35:03,182
WERE ABLE TO KILL THE

781
00:35:03,182 --> 00:35:03,483
ASPERGILLUS.

782
00:35:03,483 --> 00:35:06,452
THE CONTROL DONOR OF THE

783
00:35:06,452 --> 00:35:08,154
NEUTROPHILS IS VERY EFFECTIVE AT

784
00:35:08,154 --> 00:35:09,021
KILLING ASPERGILLUS IN THE

785
00:35:09,021 --> 00:35:11,424
PRESENCE OF NETs IF WE DESTROY

786
00:35:11,424 --> 00:35:13,926
THE NETs THEN IT'S MUCH LESS

787
00:35:13,926 --> 00:35:15,561
EFFECTIVE AS KILLING

788
00:35:15,561 --> 00:35:16,062
ASPERGILLUS.

789
00:35:16,062 --> 00:35:19,432
BEFORE GENE THERAPY THIS PATIENT

790
00:35:19,432 --> 00:35:30,009
WAS NOT ABLE , BUT AFTER ACQUIRD

791
00:35:30,777 --> 00:35:34,514
ABILITY TO KILL ASPERGILLUS.

792
00:35:34,514 --> 00:35:36,482
THESE ARE SITES OF INFECTION

793
00:35:36,482 --> 00:35:37,617
WITH ASPERGILLUS, THIS IS BEFORE

794
00:35:37,617 --> 00:35:39,118
GENE THERAPY AND TWO WEEKS AFTER

795
00:35:39,118 --> 00:35:42,755
GENE THERAPY, YOU CAN SEE THE

796
00:35:42,755 --> 00:35:43,456
INFECTION IS SOLVED.

797
00:35:43,456 --> 00:35:47,894
SO THIS IS AN EXAMPLE OF HOW

798
00:35:47,894 --> 00:35:51,964
RELEVANT OUR NETs ARE DURING

799
00:35:51,964 --> 00:35:52,398
INFECTION.

800
00:35:52,398 --> 00:35:53,032
AGAIN, STEPPING INTO WHAT

801
00:35:53,032 --> 00:35:54,734
MARIANA IS GOING TO TALK ABOUT

802
00:35:54,734 --> 00:35:57,036
NETs SEEM TO HAVE EVOLVED TO

803
00:35:57,036 --> 00:35:59,839
CONTROL INFECTIONS, AND THEY ARE

804
00:35:59,839 --> 00:36:03,342
IMPORTANT IN VIRAL BACTERIAL,

805
00:36:03,342 --> 00:36:04,944
PARASITIC, FUNGAL INFECTIONS,

806
00:36:04,944 --> 00:36:06,512
BUT AS MOST ASPECTS OF

807
00:36:06,512 --> 00:36:08,114
IMMUNOLOGY THERE'S A DARK SIDE

808
00:36:08,114 --> 00:36:12,385
OF AUTOIMMUNITY.

809
00:36:12,385 --> 00:36:12,985
THIS IS PROBABLY BECAUSE NETs

810
00:36:12,985 --> 00:36:13,686
ARE PRO-INFLAMMATORY.

811
00:36:13,686 --> 00:36:20,092
THIS IS A MOVIE OF A CULTURE

812
00:36:20,092 --> 00:36:22,829
WHERE WE MAKE MONOCYTES AND

813
00:36:22,829 --> 00:36:26,332
NEUTROPHILS, SORRY, MACROPHAGES

814
00:36:26,332 --> 00:36:28,734
AND NEUTROPHILS, THE NUCLEUS IS

815
00:36:28,734 --> 00:36:30,036
BLUE, NEUTROPHILS ARE IN PINK.

816
00:36:30,036 --> 00:36:35,074
CAN YOU SEE HOW WHEN WE ACTIVATE

817
00:36:35,074 --> 00:36:36,843
THIS CULTURE THE MACROPHAGES

818
00:36:36,843 --> 00:36:37,610
EATS THE NETs.

819
00:36:37,610 --> 00:36:43,449
SO YOU CAN FOLLOW THE  MACROPHAE

820
00:36:43,449 --> 00:36:51,457
THAT WILL EAT THE CELL, WHEN IT

821
00:36:51,457 --> 00:36:53,793
MAKES NETs.

822
00:36:53,793 --> 00:36:58,764
WE'VE SHOWN THEN NET INSIDE THE

823
00:36:58,764 --> 00:37:03,002
CELL ACTIVATE FORMATION OF THE

824
00:37:03,002 --> 00:37:05,071
TRANSCRIPTION OF CYTOKINES,

825
00:37:05,071 --> 00:37:07,273
IMPORTANTLY INTERFERON, USING

826
00:37:07,273 --> 00:37:08,808
CRISPR/Cas COULD SHOW THAT

827
00:37:08,808 --> 00:37:13,613
MACROPHAGES WHERE WE KNOCKED OUT

828
00:37:13,613 --> 00:37:17,016
EITHER CGAS OR STING, ONE OF THE

829
00:37:17,016 --> 00:37:23,422
PATHWAYS THAT NEUTROPHILS

830
00:37:23,422 --> 00:37:24,724
INTRACELLULAR AACTIVATE IS

831
00:37:24,724 --> 00:37:26,192
THROUGH CGAS AND STING.

832
00:37:26,192 --> 00:37:30,029
PATHOLOGY OF NETs IS ALSO VERY

833
00:37:30,029 --> 00:37:33,065
IMPORTANT, IT'S IMPORTANT IN

834
00:37:33,065 --> 00:37:34,400
THROMBOSIS, MANY AUTOIMMUNE

835
00:37:34,400 --> 00:37:40,072
DISEASES, AS MARIANA WILL TALK,

836
00:37:40,072 --> 00:37:41,007
ATHEROSCLEROSIS AND CANCER

837
00:37:41,007 --> 00:37:41,807
METASTASIS.

838
00:37:41,807 --> 00:37:43,442
I'LL FINISH THIS PART OF THE

839
00:37:43,442 --> 00:37:44,710
PRESENTATION BY REMINDING THAT

840
00:37:44,710 --> 00:37:49,115
YOU NEUTROPHILS WHEN THEY

841
00:37:49,115 --> 00:37:54,754
RESPOND TO MICROBES OR OTHER

842
00:37:54,754 --> 00:37:58,925
MITOGENS GO THROUGH A PROGRAMMED

843
00:37:58,925 --> 00:38:01,294
CELL DEATH, NETosis, ALLOWS

844
00:38:01,294 --> 00:38:03,162
RELEASE OF NETs, NETS CAN KILL

845
00:38:03,162 --> 00:38:07,433
MICROBES AND PROBABLY THEY

846
00:38:07,433 --> 00:38:08,367
EVOLVE, PRO-INFLAMMATORY AND PRO

847
00:38:08,367 --> 00:38:09,635
COAGULANT AND IF NETs ARE NOT

848
00:38:09,635 --> 00:38:11,037
MADE AT THE RIGHT TIME AND BY

849
00:38:11,037 --> 00:38:14,840
THE RIGHT CELLS OR THEY ARE NOT

850
00:38:14,840 --> 00:38:16,342
DESTROYED EFFICIENTLY THEY CAN

851
00:38:16,342 --> 00:38:18,311
CAUSE IMMUNITY, THERE'S MANY

852
00:38:18,311 --> 00:38:19,445
PEOPLE'S -- I PRESENTED HERE THE

853
00:38:19,445 --> 00:38:21,914
WORK OF MANY PEOPLE THAT I WILL

854
00:38:21,914 --> 00:38:26,852
LEAVE THIS SLIDE FOR A MINUTE,

855
00:38:26,852 --> 00:38:32,425
AND I WOULD LIKE TO THANK THE

856
00:38:32,425 --> 00:38:33,426
MAX PLANCK SOCIETY WHO SUPPORTED

857
00:38:33,426 --> 00:38:42,201
THIS WORK FROM THE START. 

858
00:38:42,201 --> 00:38:47,206
THANK YOU VERY MUCH.

859
00:38:47,206 --> 00:38:49,709
>> THANK YOU.

860
00:38:49,709 --> 00:39:00,152
WE'LL NOW MOVE TO MARIANA.

861
00:39:01,887 --> 00:39:02,588
>> GOOD AFTERNOON.

862
00:39:02,588 --> 00:39:04,523
I HOPE YOU CAN SEE MY SLIDES.

863
00:39:04,523 --> 00:39:05,124
>> INDEED.

864
00:39:05,124 --> 00:39:06,325
OKAY.

865
00:39:06,325 --> 00:39:06,592
WONDERFUL.

866
00:39:06,592 --> 00:39:08,160
THANK YOU.

867
00:39:08,160 --> 00:39:12,732
THANK YOU VERY MUCH, DAN, ALSO

868
00:39:12,732 --> 00:39:14,266
FOR THE KIND INTRODUCTION AND

869
00:39:14,266 --> 00:39:18,337
INVITING US TO GIVE THIS TALK, A

870
00:39:18,337 --> 00:39:24,110
PLEASURE TO BE ABLE TO GIVE THE

871
00:39:24,110 --> 00:39:25,344
PRESENTATION FOLLOWING ARTURO.

872
00:39:25,344 --> 00:39:28,514
I'M GOING TO FOCUS ON THE DARK

873
00:39:28,514 --> 00:39:30,316
SIDE OF NETs.

874
00:39:30,316 --> 00:39:34,453
AND TRY TO DISCUSS HOW WHEN

875
00:39:34,453 --> 00:39:36,422
THESE DRAMATIC AND IMPORTANT

876
00:39:36,422 --> 00:39:38,257
BIOLOGIC PROCESS OF NET

877
00:39:38,257 --> 00:39:42,662
FORMATION GOES AWRY, IT MAY

878
00:39:42,662 --> 00:39:43,996
CONTRIBUTE TO INITIATE CHRONIC

879
00:39:43,996 --> 00:39:45,965
INFLAMMATORY CONDITIONS AND I

880
00:39:45,965 --> 00:39:48,901
WILL PRIMARILY FOCUS ON SYSTEMIC

881
00:39:48,901 --> 00:39:50,302
AUTOIMMUNE DISEASES, TOWARD THE

882
00:39:50,302 --> 00:39:52,438
END OF PRESENTATION WILL TALK ON

883
00:39:52,438 --> 00:39:57,276
THE ROLE OF NETs IN PROMOTING

884
00:39:57,276 --> 00:39:59,545
VASCULAR DAMAGE, WHICH IS

885
00:39:59,545 --> 00:40:02,348
COMPLICATION THAT IS FREQUENT IN

886
00:40:02,348 --> 00:40:07,520
AUTOIMMUNE DISEASE AND OTHER

887
00:40:07,520 --> 00:40:08,521
CHRONIC INFLAMMATORY CONDITIONS.

888
00:40:08,521 --> 00:40:13,359
AND SO THIS IS A LIST OF WHAT WE

889
00:40:13,359 --> 00:40:14,527
CALL SYSTEMIC AUTOIMMUNE

890
00:40:14,527 --> 00:40:17,163
DISEASES, THEY ARE FASCINATING

891
00:40:17,163 --> 00:40:18,564
CONDITIONS, VERY CHALLENGING TO

892
00:40:18,564 --> 00:40:24,837
DIAGNOSE AND TO TREAT.

893
00:40:24,837 --> 00:40:29,308
AND ONE SHARED OBSERVATION ABOUT

894
00:40:29,308 --> 00:40:30,142
MANY SYSTEMIC AUTOIMMUNE DISEASE

895
00:40:30,142 --> 00:40:31,811
THEY ARE MUCH MORE PREVALENT IN

896
00:40:31,811 --> 00:40:32,278
WOMEN.

897
00:40:32,278 --> 00:40:36,882
HERE I'M SHOWING SOME OF THE

898
00:40:36,882 --> 00:40:38,384
ONES THAT INCLUDING SYSTEMIC

899
00:40:38,384 --> 00:40:41,721
MORE SPECIFIC ONES WHERE CAN YOU

900
00:40:41,721 --> 00:40:46,726
SEE WOMEN ARE MUCH MORE AFFECTED

901
00:40:46,726 --> 00:40:48,094
THAN MEN.

902
00:40:48,094 --> 00:40:54,233
THIS IS PROBABLY A COMBINATION

903
00:40:54,233 --> 00:40:55,801
OF DIFFERENCE IN MODULATION,

904
00:40:55,801 --> 00:40:59,371
IMMUNITY, ALSO DIFFERENCES IN

905
00:40:59,371 --> 00:41:01,741
SEX CHROMOSOMES HAVING TWO X

906
00:41:01,741 --> 00:41:05,611
CHROMOSOMES WHICH CARRY MANY

907
00:41:05,611 --> 00:41:06,679
IMMUNE-ASSOCIATED GENES MAY ALSO

908
00:41:06,679 --> 00:41:10,516
PREDISPOSE WOMEN TO HAVE MORE

909
00:41:10,516 --> 00:41:10,916
AUTOIMMUNITY.

910
00:41:10,916 --> 00:41:17,556
AND THEY ARE ALSO LONG ENCODING

911
00:41:17,556 --> 00:41:18,858
RNAs IMPLICATED IN CONDITIONS

912
00:41:18,858 --> 00:41:21,427
BY WOMEN. 

913
00:41:21,427 --> 00:41:23,996
A LOT IS KNOWN HOW SEX

914
00:41:23,996 --> 00:41:26,298
DIFFERENCES MODULATE HOW VARIOUS

915
00:41:26,298 --> 00:41:27,099
IMMUNE CELLS BEHAVE.

916
00:41:27,099 --> 00:41:29,635
WE KNEW LESS UNTIL RECENTLY

917
00:41:29,635 --> 00:41:33,272
ABOUT WHETHER THERE ARE

918
00:41:33,272 --> 00:41:34,140
SIGNIFICANT SEX DIFFERENCES IN

919
00:41:34,140 --> 00:41:39,378
NEUTROPHIL BIOLOGY, AND THIS IS

920
00:41:39,378 --> 00:41:40,846
IMPORTANT IN NET FORMATION.

921
00:41:40,846 --> 00:41:46,218
SOME YEARS AGO WE FOUND THAT

922
00:41:46,218 --> 00:41:48,420
NEUTROPHILS HAVE FEMALES AND

923
00:41:48,420 --> 00:41:51,023
MALES ARE DIFFERENT, AND WHEN

924
00:41:51,023 --> 00:41:54,026
YOU LOOK AT GENE EXPRESSION OF

925
00:41:54,026 --> 00:41:55,528
PERIPHERAL BLOOD IMMUNE CELLS

926
00:41:55,528 --> 00:41:57,263
AND COMPARE YOUNG ADULT FEMALES

927
00:41:57,263 --> 00:41:59,431
AND YOUNG ADULT MALES THE MAIN

928
00:41:59,431 --> 00:42:02,334
DIFFERENCE WE SEE IS ACTUALLY IN

929
00:42:02,334 --> 00:42:04,436
GENES ASSOCIATED TO NEUTROPHIL

930
00:42:04,436 --> 00:42:07,273
BIOLOGY AND MORE SPECIFICALLY TO

931
00:42:07,273 --> 00:42:07,573
MATURATION.

932
00:42:07,573 --> 00:42:10,176
AND LONG STORY SHORT WHAT WE

933
00:42:10,176 --> 00:42:12,311
FOUND IS THAT NEUTROPHILS IN

934
00:42:12,311 --> 00:42:14,513
WOMEN ARE MORE MATURE THAN

935
00:42:14,513 --> 00:42:17,750
NEUTROPHILS IN MEN, WHEN WE LOOK

936
00:42:17,750 --> 00:42:21,821
AT YOUNG HEALTHY ADULTS.

937
00:42:21,821 --> 00:42:27,293
AND JUST TO SUMMARIZE THE MAIN

938
00:42:27,293 --> 00:42:30,396
FINDINGS OF THESE, WOMEN HAVE

939
00:42:30,396 --> 00:42:32,998
MORE ACTIVATED NEUTROPHILS.

940
00:42:32,998 --> 00:42:34,133
THEY ACTUALLY RESPOND MORE

941
00:42:34,133 --> 00:42:35,301
ROBUSTLY TO DANGER SIGNALS.

942
00:42:35,301 --> 00:42:38,504
FOR EXAMPLE, MORE RESPONSIVE TO

943
00:42:38,504 --> 00:42:42,408
TYPE I INTERFERONS, IMPORTANT

944
00:42:42,408 --> 00:42:44,610
ANTI-VIRAL PROTEINS, ALSO HAVING

945
00:42:44,610 --> 00:42:46,245
IMPLICATION IN PATHOGENESIS OF

946
00:42:46,245 --> 00:42:47,413
AUTOIMMUNE DISEASES AND THEY ARE

947
00:42:47,413 --> 00:42:51,450
MORE LIKELY TO MAKE NETs.

948
00:42:51,450 --> 00:42:56,622
AND SO NEUTROPHILS FROM WOMEN

949
00:42:56,622 --> 00:42:59,425
UPON EXPOSURE TO INFECTIOUS

950
00:42:59,425 --> 00:43:03,495
STIMULI OR STERILE INFLAMMATORY

951
00:43:03,495 --> 00:43:04,396
STIMULI MORE ROBUSTLY FORM

952
00:43:04,396 --> 00:43:06,765
NETs, AND MAY HAVE IMPORTANT

953
00:43:06,765 --> 00:43:08,200
IMPLICATIONS FOR INFECTIONS, AS

954
00:43:08,200 --> 00:43:09,835
WE KNOW IT'S BEEN WELL DESCRIBED

955
00:43:09,835 --> 00:43:11,470
WOMEN TEND TO FIGHT INFECTIONS

956
00:43:11,470 --> 00:43:14,340
BETTER THAN MEN, BUT ALSO AS I

957
00:43:14,340 --> 00:43:19,445
MENTIONED THAT WOMEN ARE MORE

958
00:43:19,445 --> 00:43:20,980
PRONE TO AUTOIMMUNITY, ONE

959
00:43:20,980 --> 00:43:22,314
POTENTIAL FACTOR IS ACTIVATION

960
00:43:22,314 --> 00:43:25,784
STAT 

961
00:43:25,784 --> 00:43:26,886
STATUS OF NEUTROPHIL AND ABILITY

962
00:43:26,886 --> 00:43:29,421
TO FORM NETs.

963
00:43:29,421 --> 00:43:32,124
CHANGES IN MATURATION LEVEL OF

964
00:43:32,124 --> 00:43:35,561
NEUTROPHILS IN WOMEN APPEAR TO

965
00:43:35,561 --> 00:43:37,129
VARY DURING PREGNANCY WHERE

966
00:43:37,129 --> 00:43:39,465
DURING THE SECOND AND THIRD

967
00:43:39,465 --> 00:43:40,432
TRIMESTER OF PREGNANCY

968
00:43:40,432 --> 00:43:45,504
NEUTROPHILS OF WOMEN BECOME LESS

969
00:43:45,504 --> 00:43:47,539
MATURE AND RESEMBLE NEUTROPHILS

970
00:43:47,539 --> 00:43:49,475
FROM MEN, FOLLOWING POSTPARTUM

971
00:43:49,475 --> 00:43:52,811
PERIOD THEY GO BACK TO THE STAGE

972
00:43:52,811 --> 00:43:54,413
OF ENHANCED ACTIVATION AND

973
00:43:54,413 --> 00:43:55,581
MATURATION.

974
00:43:55,581 --> 00:43:57,483
SO WE ARE HYPOTHESIZING, THIS IS

975
00:43:57,483 --> 00:43:59,451
WORK ONGOING, THAT SEX HORMONES

976
00:43:59,451 --> 00:44:02,488
ARE PROBABLY PLAYING AN

977
00:44:02,488 --> 00:44:04,256
IMPORTANT ROLE IN THE NEUTROPHIL

978
00:44:04,256 --> 00:44:05,124
BIOLOGY, FOR INSTANCE.

979
00:44:05,124 --> 00:44:06,659
THIS LEADS ME TO THE NEXT PART

980
00:44:06,659 --> 00:44:10,963
OF THE TALK, WHICH IS I WANTED

981
00:44:10,963 --> 00:44:13,632
TO START WITH THIS PREMISE THAT

982
00:44:13,632 --> 00:44:15,200
AUTOIMMUNE DISEASE IS MORE

983
00:44:15,200 --> 00:44:17,069
COMMON IN WOMEN.

984
00:44:17,069 --> 00:44:20,239
WOMEN NEUTROPHIL APPEARS TO FORM

985
00:44:20,239 --> 00:44:27,579
MORE  NETs, WHAT IS THE ROLE IN

986
00:44:27,579 --> 00:44:29,248
AUTOIMMUNITY?

987
00:44:29,248 --> 00:44:30,716
AUTOIMMUNITY DISEASES ARE

988
00:44:30,716 --> 00:44:33,252
HETEROGENEOUS, THERE ARE

989
00:44:33,252 --> 00:44:35,354
COMMONALITIES, AS MANY CHRONIC

990
00:44:35,354 --> 00:44:36,522
CONDITIONS WE CONSIDER THEM

991
00:44:36,522 --> 00:44:38,824
MULTI-STEP PROCESSES WHERE BOTH

992
00:44:38,824 --> 00:44:39,358
GENETIC AND ENVIRONMENTAL

993
00:44:39,358 --> 00:44:42,995
FACTORS PLAY KEY ROLES IN

994
00:44:42,995 --> 00:44:44,530
INDUCING CLINICAL STATES WHERE

995
00:44:44,530 --> 00:44:47,433
BOTH INNATE AND ADAPTIVE IMMUNE

996
00:44:47,433 --> 00:44:49,768
RESPONSES BECOME VERY ABNORMAL.

997
00:44:49,768 --> 00:44:52,271
AND THROUGH A VARIETY OF

998
00:44:52,271 --> 00:44:52,871
PROGRAMS THERE'S ENVIRONMENTAL

999
00:44:52,871 --> 00:45:00,646
FACTORS AND  STOCHASTIC EVENTS,

1000
00:45:00,646 --> 00:45:02,614
RECEIVING VARIOUS TIPS FROM THE

1001
00:45:02,614 --> 00:45:05,017
ENVIRONMENT THAT WOULD LEAD

1002
00:45:05,017 --> 00:45:07,386
EVENTUALLY TO PRODUCTION OF

1003
00:45:07,386 --> 00:45:11,790
AUTOANTIBODIES THAT TARGET

1004
00:45:11,790 --> 00:45:12,591
CELLULAR AND EXTRACELLULAR

1005
00:45:12,591 --> 00:45:12,958
AUTOANTIGENS.

1006
00:45:12,958 --> 00:45:16,195
INDIVIDUALS AT RISK MAY DEVELOP

1007
00:45:16,195 --> 00:45:18,230
THESE AUTOANTIBODIES MANY YEARS

1008
00:45:18,230 --> 00:45:21,967
BEFORE CLINICAL DISEASE DEVELOPS

1009
00:45:21,967 --> 00:45:23,302
BUT EVENTUALLY AFTER ADDITIONAL

1010
00:45:23,302 --> 00:45:26,672
HITS FROM THE ENVIRONMENT AN

1011
00:45:26,672 --> 00:45:27,573
INDIVIDUAL WILL DEVELOP

1012
00:45:27,573 --> 00:45:28,240
CHARACTERISTICS OF SYSTEMIC

1013
00:45:28,240 --> 00:45:31,744
AUTOIMMUNE  DISEASE, GO THROUGH

1014
00:45:31,744 --> 00:45:33,245
REMISSION AND FLARES, AS THEY

1015
00:45:33,245 --> 00:45:35,781
LIVE LONGER NOW WE KNOW THEY

1016
00:45:35,781 --> 00:45:37,783
HAVE A HIGHER RISK TO ACQUIRE

1017
00:45:37,783 --> 00:45:41,487
CONDITIONS THAT ARE KNOWN TO BE

1018
00:45:41,487 --> 00:45:43,355
SECONDARY TO PROLONGED EXPOSURE

1019
00:45:43,355 --> 00:45:46,492
TO OXIDATIVE STRESS, SUCH AS

1020
00:45:46,492 --> 00:45:47,092
CARDIOVASCULAR DISEASE, BONE

1021
00:45:47,092 --> 00:45:49,561
DAMAGE, FIBROSIS, AND CANCER.

1022
00:45:49,561 --> 00:45:52,865
AND SO FOR US STUDYING

1023
00:45:52,865 --> 00:45:56,101
AUTOIMMUNITY ONE KEY QUESTION

1024
00:45:56,101 --> 00:46:00,005
HAS BEEN HOW DO THESE

1025
00:46:00,005 --> 00:46:07,913
INTRACELLULAR ANTIGENS BECOME

1026
00:46:07,913 --> 00:46:09,515
IMMUNOGENS, THE DOGMA FOR MANY

1027
00:46:09,515 --> 00:46:15,020
YEARS HAS BEEN THERE'S A

1028
00:46:15,020 --> 00:46:17,322
DYSREGULATION IN CELL DEATH

1029
00:46:17,322 --> 00:46:19,158
FOCUSED ON APOPTOSIS, INCREASE

1030
00:46:19,158 --> 00:46:21,160
THE APOPTOSIS OR INCREASED CELL

1031
00:46:21,160 --> 00:46:23,562
DEATH OR CLEARANCE OF THE DEAD

1032
00:46:23,562 --> 00:46:27,266
CELLS MAY PROMOTE TO INCREASING

1033
00:46:27,266 --> 00:46:28,267
HALF-LIFE OF AUTOANTIGENS AND

1034
00:46:28,267 --> 00:46:31,136
MODIFYING AUTOANTIGENS IN A WAY

1035
00:46:31,136 --> 00:46:33,839
THAT PRE-DISPOSED INDIVIDUALS

1036
00:46:33,839 --> 00:46:38,877
MAY LEAD TO AUTOANTIBODY

1037
00:46:38,877 --> 00:46:40,579
FORMATION AND OUR DOWNSTREAM

1038
00:46:40,579 --> 00:46:40,979
PROCESSES.

1039
00:46:40,979 --> 00:46:42,014
WE FOCUSED ON APOPTOSIS BECAUSE

1040
00:46:42,014 --> 00:46:43,649
WE KNOW LESS ABOUT OTHER TIMES

1041
00:46:43,649 --> 00:46:46,351
OF CELL DEATH, WE NOW KNOW CELLS

1042
00:46:46,351 --> 00:46:48,120
CAN DIE IN MANY DIFFERENT WAYS.

1043
00:46:48,120 --> 00:46:53,892
NOT ONLY WHETHER A CELL WILL DIE

1044
00:46:53,892 --> 00:46:58,497
OR NOT DIE BUT HOW, WHICH TYPE,

1045
00:46:58,497 --> 00:47:09,041
WHAT IS THE MICROENVIRONMENT, TO

1046
00:47:10,609 --> 00:47:11,376
DAMPEN INFLAMMATORY RESPONSE.

1047
00:47:11,376 --> 00:47:14,413
AND SO WHEN ARTURO PUBLISHED HIS

1048
00:47:14,413 --> 00:47:16,949
INITIAL WORK ON NETs, WE

1049
00:47:16,949 --> 00:47:19,151
BECAME EXCITED BECAUSE THIS WAS

1050
00:47:19,151 --> 00:47:21,086
A NOVEL CELL DEATH MECHANISM

1051
00:47:21,086 --> 00:47:23,455
THAT MADE A LOT OF SENSE, COULD

1052
00:47:23,455 --> 00:47:25,557
PLAY POTENTIAL ROLE IN

1053
00:47:25,557 --> 00:47:25,891
AUTOIMMUNITY.

1054
00:47:25,891 --> 00:47:29,495
AND SO WE NOW KNOW THAT FROM THE

1055
00:47:29,495 --> 00:47:30,596
INITIAL DESCRIPTION THAT

1056
00:47:30,596 --> 00:47:31,630
MICROBES ARE MAIN INDUCERS OF

1057
00:47:31,630 --> 00:47:34,766
NET FORMATION THERE ARE A NUMBER

1058
00:47:34,766 --> 00:47:35,868
OF OTHER STERILE INFLAMMATORY

1059
00:47:35,868 --> 00:47:39,638
STOMACH LIE -- STIMULI WITH

1060
00:47:39,638 --> 00:47:42,141
ABILITY TO INDUCE NET FORMATION

1061
00:47:42,141 --> 00:47:44,543
INCLUDING PLATELETS, CYTOKINES,

1062
00:47:44,543 --> 00:47:49,815
COMPLEMENT COMPONENTS, VARIOUS

1063
00:47:49,815 --> 00:47:52,618
CRYSTALS, URIC ACID CRYSTALS AND

1064
00:47:52,618 --> 00:47:54,953
COMPONENTS THAT ARE IMPORTANT IN

1065
00:47:54,953 --> 00:47:57,456
AUTOIMMUNE DISEASES SUCH AS

1066
00:47:57,456 --> 00:47:57,890
IMMUNE COMPLEXES AND

1067
00:47:57,890 --> 00:47:59,224
AUTOANTIBODIES, ALL OF THESE MAY

1068
00:47:59,224 --> 00:48:03,162
LED TO FORMATION OF NETs, VERY

1069
00:48:03,162 --> 00:48:06,331
IMPORTANTLY AS ARTURO MENTIONED

1070
00:48:06,331 --> 00:48:10,402
DURING HIS TALK THAT NETs

1071
00:48:10,402 --> 00:48:12,671
CARRY OR EXTERNALIZE MANY

1072
00:48:12,671 --> 00:48:15,440
AUTOANTIGENS KNOWN TO BE KEY

1073
00:48:15,440 --> 00:48:15,707
TARGETS.

1074
00:48:15,707 --> 00:48:21,113
IN THE CASE OF LUPUS, FOR

1075
00:48:21,113 --> 00:48:24,917
EXAMPLE, IN OTHER AUTOIMMUNE

1076
00:48:24,917 --> 00:48:25,317
CONDITIONS

1077
00:48:25,317 --> 00:48:29,555
IN THE KALES OF RHEUMATOID

1078
00:48:29,555 --> 00:48:30,656
ARTHRITIS MODIFIED PROTEINS

1079
00:48:30,656 --> 00:48:32,090
EXTERNALIZED WILL BE KEY

1080
00:48:32,090 --> 00:48:33,392
AUTOANTIGENS IN THE DISEASE.

1081
00:48:33,392 --> 00:48:36,195
AND WE NOW KNOW THAT FOR MANY

1082
00:48:36,195 --> 00:48:39,531
AUTOIMMUNE DISEASES THERE'S A

1083
00:48:39,531 --> 00:48:41,366
PROFOUND IMBALANCE BETWEEN NET

1084
00:48:41,366 --> 00:48:44,536
FORMATION INCREASED AND NET

1085
00:48:44,536 --> 00:48:46,505
DEGRADATION THAT IS IMPAIRED,

1086
00:48:46,505 --> 00:48:50,175
STARTING WITH ARTURO'S GROUPS

1087
00:48:50,175 --> 00:48:52,177
AND VARIOUS GROUPS HAVE SHOWN A

1088
00:48:52,177 --> 00:48:55,447
THIRD OF LUPUS PATIENTS HAVE

1089
00:48:55,447 --> 00:48:58,984
IMPAIRMENT IN NET DEGRADATION,

1090
00:48:58,984 --> 00:48:59,751
HALF-LIFE MAY BE INCREASED IN

1091
00:48:59,751 --> 00:49:01,853
CIRCULATION AND ALSO IN TISSUES.

1092
00:49:01,853 --> 00:49:04,756
WE HAVE FOUND THAT THIS

1093
00:49:04,756 --> 00:49:06,725
PHENOMENON IS NOT ONLY IN VITRO

1094
00:49:06,725 --> 00:49:09,361
BUT OPERATIONALLY IN VIVO

1095
00:49:09,361 --> 00:49:10,596
BECAUSE IN MANY AUTOIMMUNE

1096
00:49:10,596 --> 00:49:14,266
DISEASES WE FOUND EVIDENCE OF

1097
00:49:14,266 --> 00:49:15,200
NETTING NEUTROPHILS IN THE

1098
00:49:15,200 --> 00:49:16,868
TISSUES OF THESE PATIENTS.

1099
00:49:16,868 --> 00:49:19,605
AND SO I'M GOING TO TELL BUT

1100
00:49:19,605 --> 00:49:21,640
THREE AUTOIMMUNE CONDITIONS AND

1101
00:49:21,640 --> 00:49:24,076
THE WORK WE'VE DONE TO SHOW

1102
00:49:24,076 --> 00:49:26,712
EVIDENCE THAT NETs MAY BE

1103
00:49:26,712 --> 00:49:29,147
IMPLICATED IN THE DEVELOPMENT

1104
00:49:29,147 --> 00:49:30,048
AND PERPETRATION OF AUTOIMMUNE

1105
00:49:30,048 --> 00:49:30,449
DISEASES.

1106
00:49:30,449 --> 00:49:32,784
ONE WHERE WE CAN FOCUSED MOST OF

1107
00:49:32,784 --> 00:49:41,693
THE RESOURCE IS SYSTEMIC LUPUS

1108
00:49:41,693 --> 00:49:41,827
,

1109
00:49:41,827 --> 00:49:46,565
THE POSTER CHILD OF SYSTEMIC

1110
00:49:46,565 --> 00:49:46,898
AUTOIMMUNITY.

1111
00:49:46,898 --> 00:49:49,868
AFFECTS PRIMARILY WOMEN, IT IS

1112
00:49:49,868 --> 00:49:50,369
PROFOUNDLY HETEROGENEOUS

1113
00:49:50,369 --> 00:49:51,069
CONDITION, MORE A SYNDROME THAN

1114
00:49:51,069 --> 00:49:52,504
A DISEASE.

1115
00:49:52,504 --> 00:49:54,873
THERE ARE PROBLEM MANY WAYS BY

1116
00:49:54,873 --> 00:49:57,509
WHICH WE CAN -- AN INDIVIDUAL

1117
00:49:57,509 --> 00:50:01,113
CAN DEVELOP LUPUS.

1118
00:50:01,113 --> 00:50:01,913
CHARACTERISTICSALLY THE SINE QUA

1119
00:50:01,913 --> 00:50:05,083
NON IS DEVELOPMENT OF

1120
00:50:05,083 --> 00:50:06,485
ANTIBODIES, TARGETS NUCLEIC ACID

1121
00:50:06,485 --> 00:50:09,187
OR PROTEINS BINDING SUCH AS

1122
00:50:09,187 --> 00:50:10,155
HISTONES, SO TO THE NUCLEOSOME.

1123
00:50:10,155 --> 00:50:13,825
IF WE DO NOT SEE THESE WE DOUBT

1124
00:50:13,825 --> 00:50:18,130
A PATIENT WOULD HAVE LUPUS.

1125
00:50:18,130 --> 00:50:20,032
THESE PATIENTS HAVE PROFOUND

1126
00:50:20,032 --> 00:50:20,966
DYSREGULATION IN THE TYPE I

1127
00:50:20,966 --> 00:50:23,935
INTERFERON PATHWAY THAT I

1128
00:50:23,935 --> 00:50:26,238
MENTIONED TO YOU, HOST DEFENSE,

1129
00:50:26,238 --> 00:50:30,275
ALSO IN AUTOIMMUNITY

1130
00:50:30,275 --> 00:50:32,444
DEVELOPMENT, SIGNIFICANT

1131
00:50:32,444 --> 00:50:33,445
PROPORTION OF LUPUS PATIENTS

1132
00:50:33,445 --> 00:50:35,414
HAVE ENHANCED RISK TO DEVELOP

1133
00:50:35,414 --> 00:50:39,017
HEART ATTACKS OR STROKE AND WE

1134
00:50:39,017 --> 00:50:42,621
KNOW THIS IS PRIMARILY DRIVEN BY

1135
00:50:42,621 --> 00:50:44,589
IMMUNE DYSREGULATION

1136
00:50:44,589 --> 00:50:45,957
CHARACTERISTICS OF LUPUS.

1137
00:50:45,957 --> 00:50:49,361
AND SO ALMOST 15 YEARS AGO WE

1138
00:50:49,361 --> 00:50:53,231
FOUND THAT WHEN WE WERE TRYING

1139
00:50:53,231 --> 00:50:56,668
TO PURIFY MONO NUCLEAR CELLS

1140
00:50:56,668 --> 00:50:58,937
FOLLOWING GRADIENT, EVEN THE

1141
00:50:58,937 --> 00:51:02,240
INCREASED DENSITY IN REGULAR

1142
00:51:02,240 --> 00:51:03,475
NEUTROPHILS TYPICALLY WHEN WE DO

1143
00:51:03,475 --> 00:51:05,344
THIS GO MORE TO THE BOTTOM NEXT

1144
00:51:05,344 --> 00:51:09,715
TO RED BLOOD CELLS AND THEN MONO

1145
00:51:09,715 --> 00:51:11,616
NUCLEAR CELLS ARE ISOLATED

1146
00:51:11,616 --> 00:51:13,685
HIGHER UP IN THESE WHAT WE CALL

1147
00:51:13,685 --> 00:51:14,586
MONO NUCLEAR CELL ENTER PHASE.

1148
00:51:14,586 --> 00:51:20,859
IN THE CASE OF LUPUS WHEN TRYING

1149
00:51:20,859 --> 00:51:23,095
TO PURIFY WERE ALWAYS FINDING

1150
00:51:23,095 --> 00:51:25,964
SIGNIFICANT PROPORTION OF

1151
00:51:25,964 --> 00:51:29,101
NEUTROPHILS PRESENT IN THESE

1152
00:51:29,101 --> 00:51:31,169
MONONUCLEAR CELL INTERFACE.

1153
00:51:31,169 --> 00:51:33,605
WHAT WE FOUND LUPUS PATIENTS HAD

1154
00:51:33,605 --> 00:51:37,442
A SUBSET OF NEUTROPHILS THAT WE

1155
00:51:37,442 --> 00:51:38,777
NAMED LOW DENSITY GRANULOCYTES

1156
00:51:38,777 --> 00:51:42,848
BECAUSE THEY SEEM TO FLOAT SUCH

1157
00:51:42,848 --> 00:51:44,916
AS THEY HAD LOWER THAN REGULAR

1158
00:51:44,916 --> 00:51:47,018
NEUTROPHILS, AND WE FOUND ONE OF

1159
00:51:47,018 --> 00:51:49,755
THE KEY FEATURES OF THESE LOW

1160
00:51:49,755 --> 00:51:52,124
DENSITY GRANULOCYTES OR

1161
00:51:52,124 --> 00:51:53,658
NEUTROPHILS THAT WERE PURIFIED

1162
00:51:53,658 --> 00:51:55,193
FROM LUPUS PATIENTS IN SOME

1163
00:51:55,193 --> 00:52:00,665
CASES REPRESENTING UP TO 50%,

1164
00:52:00,665 --> 00:52:02,000
THEY CAME OUT OF CIRCULATION

1165
00:52:02,000 --> 00:52:03,201
PRIMED TO FORM NETs.

1166
00:52:03,201 --> 00:52:04,703
THERE WAS NOTHING WE COULD

1167
00:52:04,703 --> 00:52:07,439
FURTHER DO IN VITRO TO STIMULATE

1168
00:52:07,439 --> 00:52:07,939
NET FORMATION.

1169
00:52:07,939 --> 00:52:09,775
THEY WERE ALREADY MAXED OUT IN

1170
00:52:09,775 --> 00:52:12,043
THEIR ABILITY TO FORM NETs.

1171
00:52:12,043 --> 00:52:13,912
AND WE HAVE STUDIED THESE CELLS

1172
00:52:13,912 --> 00:52:15,547
OVER MANY YEARS TO TRY TO FIGURE

1173
00:52:15,547 --> 00:52:18,216
OUT FIRST WHERE ARE THEY COMING

1174
00:52:18,216 --> 00:52:21,353
FROM, SECOND WHAT IS THEIR ROLE.

1175
00:52:21,353 --> 00:52:25,190
ARE THEY JUST A PHENOMENON,

1176
00:52:25,190 --> 00:52:25,657
INNOCENT BYSTANDERS?

1177
00:52:25,657 --> 00:52:27,192
DO THEY REPRESENT MERELY THE

1178
00:52:27,192 --> 00:52:28,693
GRANULATED CELLS?

1179
00:52:28,693 --> 00:52:33,098
AND WE HAVE COME AFTER MANY

1180
00:52:33,098 --> 00:52:38,670
YEARS OF STUDY THESE APPEAR TO

1181
00:52:38,670 --> 00:52:40,338
REPRESENT PRO-INFLAMMATORY

1182
00:52:40,338 --> 00:52:43,074
SUBSET IN LUPUS PATIENTS ENDOWED

1183
00:52:43,074 --> 00:52:44,476
WITH PATHOGENIC CAPACITY.

1184
00:52:44,476 --> 00:52:46,545
AMONG THEM LISTING HERE SOME OF

1185
00:52:46,545 --> 00:52:47,779
THE ONES WE'VE FOUND.

1186
00:52:47,779 --> 00:52:51,850
THEY HAVE A HIGHER ABILITY TO BE

1187
00:52:51,850 --> 00:52:53,852
PRO-INFLAMMATORY, TO SYNTHESIZE

1188
00:52:53,852 --> 00:52:55,654
FROM CYTOKINES, A KEY FEATURE IS

1189
00:52:55,654 --> 00:52:57,856
THAT THEY CAN MAKE NETs

1190
00:52:57,856 --> 00:53:00,058
WITHOUT ANY IN VITRO

1191
00:53:00,058 --> 00:53:00,459
STIMULATION.

1192
00:53:00,459 --> 00:53:01,927
SOMETHING HAS HAPPENED IN VIVO

1193
00:53:01,927 --> 00:53:05,664
THAT MADE THEM VERY PRONE TO

1194
00:53:05,664 --> 00:53:06,331
FORM NETs.

1195
00:53:06,331 --> 00:53:07,933
NETs HAVE ABILITY TO DAMAGE

1196
00:53:07,933 --> 00:53:11,002
ENDOTHELIAL CELLS, A KEY FEATURE

1197
00:53:11,002 --> 00:53:13,338
OF THESE GRANULOCYTES, THEY ARE

1198
00:53:13,338 --> 00:53:14,673
NOT MYELOID SUPPRESSOR CELLS.

1199
00:53:14,673 --> 00:53:16,408
THEY HAVE STRONG ABILITY TO

1200
00:53:16,408 --> 00:53:18,076
ACTIVATE T CELLS, FURTHER

1201
00:53:18,076 --> 00:53:19,110
PROMOTING INFLAMMATION.

1202
00:53:19,110 --> 00:53:22,447
AND SOMETHING QUITE UNIQUE ABOUT

1203
00:53:22,447 --> 00:53:24,516
THE CELLS THEIR BIOMECHANICAL

1204
00:53:24,516 --> 00:53:25,917
PROPERTIES APPEAR TO BE

1205
00:53:25,917 --> 00:53:27,052
DIFFERENT THAN REGULAR

1206
00:53:27,052 --> 00:53:29,654
NEUTROPHILS, IF WE MAKE THEM

1207
00:53:29,654 --> 00:53:32,657
TRAVEL TO SOME SORT OF SMALL

1208
00:53:32,657 --> 00:53:34,593
BLOOD VESSEL IN EPIC THEY GET

1209
00:53:34,593 --> 00:53:36,461
TRAPPED IN THESE SMALL BLOOD

1210
00:53:36,461 --> 00:53:39,965
VESSELS COMPARED TO NORMAL

1211
00:53:39,965 --> 00:53:41,366
DENSITY NEUTROPHILS SUGGESTING

1212
00:53:41,366 --> 00:53:43,435
SEEM TO BE STIFFER, ADAPT LESS

1213
00:53:43,435 --> 00:53:46,304
WELL TO TRAVELING IN SMALL BLOOD

1214
00:53:46,304 --> 00:53:49,608
VESSELS AND MAY PROMOTE FOR

1215
00:53:49,608 --> 00:53:50,575
EXAMPLE TISSUE HYPERPERFUSION

1216
00:53:50,575 --> 00:53:53,645
AND ORGAN DAMAGE AND SEEM TO

1217
00:53:53,645 --> 00:53:54,846
HAVE DIFFERENT BIOENERGETICS

1218
00:53:54,846 --> 00:53:56,081
THAN REGULAR NEUTROPHILS.

1219
00:53:56,081 --> 00:53:58,884
FURTHERMORE AS WE HAVE STUDIED

1220
00:53:58,884 --> 00:54:00,185
OTHER AUTOIMMUNE DISEASES AND

1221
00:54:00,185 --> 00:54:01,286
OTHER AUTO-INFLAMMATORY DISEASES

1222
00:54:01,286 --> 00:54:02,521
WE'VE BEEN ABLE TO STUDY HERE AT

1223
00:54:02,521 --> 00:54:06,358
THE NIH WE HAVE FOUND THAT IN

1224
00:54:06,358 --> 00:54:07,425
OTHER AUTOIMMUNE INFLAMMATORY

1225
00:54:07,425 --> 00:54:08,660
CONDITIONS THERE'S ALSO THIS

1226
00:54:08,660 --> 00:54:10,362
PRESENCE OF LOW DENSITY

1227
00:54:10,362 --> 00:54:11,296
GRANULOCYTES AND INCREASED

1228
00:54:11,296 --> 00:54:13,164
NUMBERS THAT HAVE AN ENHANCED

1229
00:54:13,164 --> 00:54:15,166
CAPACITY TO FORM THESE NETs.

1230
00:54:15,166 --> 00:54:16,234
AND SOMETHING THAT'S PERHAPS

1231
00:54:16,234 --> 00:54:18,904
SHAREDDED BY MANY OF THESE

1232
00:54:18,904 --> 00:54:27,579
AUTOIMMUNE AND AUTO-INFLAMMATORY

1233
00:54:27,579 --> 00:54:30,682
CONDITIONS IS ABILITY TO FORM

1234
00:54:30,682 --> 00:54:32,317
DYSREGULATED NETs, MAY LEAD A

1235
00:54:32,317 --> 00:54:35,453
SHARED PHENOTYPE TO ENHANCE

1236
00:54:35,453 --> 00:54:41,059
BLOOD VESSEL DAMAGE TO PROMOTE

1237
00:54:41,059 --> 00:54:42,827
END ORGAN COMPLICATIONS AND

1238
00:54:42,827 --> 00:54:43,461
PREMATURE DISEASE.

1239
00:54:43,461 --> 00:54:50,302
SO, THIS IS THE WORK SUMMARIZING

1240
00:54:50,302 --> 00:54:52,671
OF OTHER GROUPS IT APPEARS WHEN

1241
00:54:52,671 --> 00:54:54,472
THE PROCESS OF NET FORMATION

1242
00:54:54,472 --> 00:54:59,611
GOES AWRY IT CONTRIBUTES TO AN

1243
00:54:59,611 --> 00:55:04,616
OF A BE IN -- A NUMBER OF

1244
00:55:04,616 --> 00:55:05,817
SYNDROMES.

1245
00:55:05,817 --> 00:55:09,621
WE KNOW NETs ARE IMPLICATED IN

1246
00:55:09,621 --> 00:55:11,923
DEVELOPMENT OF ATHEROSCLEROSIS

1247
00:55:11,923 --> 00:55:15,193
AND THROMBOSIS, CANCER AND

1248
00:55:15,193 --> 00:55:16,094
METASTASES, METABOLIC SYNDROME,

1249
00:55:16,094 --> 00:55:18,029
DIABETES, THINGS AS MUNDANE OR

1250
00:55:18,029 --> 00:55:19,698
COMMON AS GALLBLADDER STONES AND

1251
00:55:19,698 --> 00:55:26,204
MORE RECENTLY THEY WERE ALSO

1252
00:55:26,204 --> 00:55:30,442
IMPLICATED IN COMPLICATIONS IN

1253
00:55:30,442 --> 00:55:33,478
THE COVID-19 PANDEMIC.

1254
00:55:33,478 --> 00:55:39,050
FORMING NETs IN CONTEXT OF

1255
00:55:39,050 --> 00:55:41,186
IMMUNITY LEAD TO VICIOUS CYCLE

1256
00:55:41,186 --> 00:55:43,021
PROMOTING INFLAMMATION IN ORGAN

1257
00:55:43,021 --> 00:55:46,091
DAMAGE SUCH AS, FOR EXAMPLE,

1258
00:55:46,091 --> 00:55:47,626
INDUCING INFLAMMASOME ACTIVATION

1259
00:55:47,626 --> 00:55:49,294
IN MACROPHAGES, PROMOTING

1260
00:55:49,294 --> 00:55:53,198
ACTIVATION OF T CELLS AND B

1261
00:55:53,198 --> 00:55:54,399
CELLS WITH AUTOANTIBODY

1262
00:55:54,399 --> 00:55:56,034
PROTECTION, LEADING THE KEY

1263
00:55:56,034 --> 00:55:57,369
CELLS PRODUCING TYPE I

1264
00:55:57,369 --> 00:56:00,705
INTERFERONS TO MAKE MORE OF IT

1265
00:56:00,705 --> 00:56:07,412
WHICH IS IMPORTANT IMPLICATION

1266
00:56:07,412 --> 00:56:08,013
IN AUTOIMMUNITY.

1267
00:56:08,013 --> 00:56:10,849
NETs ARE MORE PRONE TO PROMOTE

1268
00:56:10,849 --> 00:56:13,585
VASCULAR DAMAGE AND THROMBOSIS.

1269
00:56:13,585 --> 00:56:15,453
AND SO WHY THE KEY QUESTION,

1270
00:56:15,453 --> 00:56:17,589
SOMETHING WE HAVE ASKED FOR A

1271
00:56:17,589 --> 00:56:28,099
NUMBER OF YEARS, ARE ALL NETs

1272
00:56:29,834 --> 00:56:30,468
CREATED EQUAL?

1273
00:56:30,468 --> 00:56:33,872
AND WHAT WE THINK IS ACTUALLY

1274
00:56:33,872 --> 00:56:36,941
NOT ALL NETs ARE CREATED THE

1275
00:56:36,941 --> 00:56:37,509
SAME.

1276
00:56:37,509 --> 00:56:39,344
AFTER VARIOUS WAYS BY WHICH WE

1277
00:56:39,344 --> 00:56:41,212
HAVE LOOKED AT THIS, FOR EXAMPLE

1278
00:56:41,212 --> 00:56:45,250
WE KNOW THAT IF NETs ARE

1279
00:56:45,250 --> 00:56:47,352
INDUCED THROUGH LUPUS RELEVANT

1280
00:56:47,352 --> 00:56:57,729
STIMULI SUCH AS IMMUNE

1281
00:56:59,497 --> 00:57:00,699
COMPLEXES, ENHANCED FORMATIONS

1282
00:57:00,699 --> 00:57:03,101
NOT ONLY IT'S IMPORTANT IN

1283
00:57:03,101 --> 00:57:05,837
GENOMIC DNA BUT MAY ALSO LEAD TO

1284
00:57:05,837 --> 00:57:07,439
ENHANCED OXIDATION OF

1285
00:57:07,439 --> 00:57:08,039
MITOCHONDRIAL DNA.

1286
00:57:08,039 --> 00:57:10,542
AND SO IN AUTOIMMUNE DISEASES WE

1287
00:57:10,542 --> 00:57:14,746
HAVE FOUND THAT THE NETs ARE

1288
00:57:14,746 --> 00:57:16,614
EXCLUDED IN OXIDATION OF NUCLEIC

1289
00:57:16,614 --> 00:57:17,916
ACID PARTICULARLY MITOCHONDRIAL

1290
00:57:17,916 --> 00:57:21,086
DNA BUT ALSO GENOMIC DNA.

1291
00:57:21,086 --> 00:57:24,155
AND AS MENTIONED BY ARTURO WE

1292
00:57:24,155 --> 00:57:26,324
HAVE FOUND ALSO THAT WHEN THESE

1293
00:57:26,324 --> 00:57:29,394
NETs ARE INTERNALIZED BY

1294
00:57:29,394 --> 00:57:30,662
TARGET CELLS, THEIR NUCLEIC ACID

1295
00:57:30,662 --> 00:57:33,932
IS LESS LIKELY TO BE DEGRADED

1296
00:57:33,932 --> 00:57:40,538
BECAUSE IT'S OXIDIZED, SO

1297
00:57:40,538 --> 00:57:41,573
EXTRACELLULARLY AND

1298
00:57:41,573 --> 00:57:49,814
INTRACELLULARLY HAVE A LONGER

1299
00:57:49,814 --> 00:57:56,421
HALF-LIFE, TRIGGERING CGAS

1300
00:57:56,421 --> 00:57:59,257
PATHWAY RELEVANT IN

1301
00:57:59,257 --> 00:58:00,291
AUTOIMMUNITY.

1302
00:58:00,291 --> 00:58:02,293
NOT ONLY NUCLEIC ACID, GENOMIC

1303
00:58:02,293 --> 00:58:05,563
DNA, MITOCHONDRIAL AND RNA ARE

1304
00:58:05,563 --> 00:58:08,967
OXIDIZED AND PRO IMMUNOGENIC BUT

1305
00:58:08,967 --> 00:58:12,137
THE PROTEIN CONTENT, THE CARGO

1306
00:58:12,137 --> 00:58:13,671
AND POST-TRANSLATIONAL

1307
00:58:13,671 --> 00:58:15,440
MODIFICATIONS PRESENT IN NETs

1308
00:58:15,440 --> 00:58:17,609
MAY VARY DEPENDING ON THE

1309
00:58:17,609 --> 00:58:18,676
STIMULUS AND EXAMINE

1310
00:58:18,676 --> 00:58:19,043
IMMUNOGENICITY.

1311
00:58:19,043 --> 00:58:19,944
I'LL TELL YOU MORE ABOUT THIS

1312
00:58:19,944 --> 00:58:22,747
LATER ON DURING MY TALK.

1313
00:58:22,747 --> 00:58:24,315
BOTH PROTEINS AND NUCLEIC ACID

1314
00:58:24,315 --> 00:58:25,383
ARE DIFFERENT DEPENDING HOW

1315
00:58:25,383 --> 00:58:28,052
NETs ARE FORMED AND THIS IS

1316
00:58:28,052 --> 00:58:30,021
WHY NOT JUST EVERYONE IS FORMING

1317
00:58:30,021 --> 00:58:32,257
NETs MAY BE CONDUCIVE TO

1318
00:58:32,257 --> 00:58:40,298
DEVELOP AUTOIMMUNITY.

1319
00:58:40,298 --> 00:58:41,166
ANOTHER IMPORTANT FACTOR THAT

1320
00:58:41,166 --> 00:58:43,034
MAY BE IMPORTANT IN DEVELOPMENT

1321
00:58:43,034 --> 00:58:45,270
OF AUTOIMMUNE DISEASES AND ORGAN

1322
00:58:45,270 --> 00:58:47,872
DAMAGE IS NETs ARE CARRIERS OF

1323
00:58:47,872 --> 00:58:50,875
A NUMBER OF MESSENGERS THAT

1324
00:58:50,875 --> 00:58:53,144
DEPENDING ON THE CELL IN THE

1325
00:58:53,144 --> 00:58:55,446
TISSUE BEING AN IMMUNE CELL OR

1326
00:58:55,446 --> 00:58:56,781
STROMAL CELL, MAY INDUCE

1327
00:58:56,781 --> 00:58:58,850
DOWNSTREAM RESPONSES THAT MAY BE

1328
00:58:58,850 --> 00:58:59,918
PRO-INFLAMMATORY.

1329
00:58:59,918 --> 00:59:02,053
AN EXAMPLE HERE IS THAT NETs

1330
00:59:02,053 --> 00:59:05,423
CARRY SMALL RNAs, AND WE HAVE

1331
00:59:05,423 --> 00:59:07,792
FOUND THESE MAY ACTIVATE IN THE

1332
00:59:07,792 --> 00:59:10,695
CELLS TO HAVE PRO-INFLAMMATORY

1333
00:59:10,695 --> 00:59:11,462
RESPONSES.

1334
00:59:11,462 --> 00:59:17,101
NETs HAVE ALSO BEEN FOUND

1335
00:59:17,101 --> 00:59:18,536
THROUGH MODIFIED HISTONES TO

1336
00:59:18,536 --> 00:59:22,273
INDUCE DIRECT DAMAGE TO SKELETAL

1337
00:59:22,273 --> 00:59:23,474
MUSCLE, IMPLICATED IN

1338
00:59:23,474 --> 00:59:25,143
PATHOGENESIS OF ANOTHER

1339
00:59:25,143 --> 00:59:28,746
AUTOIMMUNE DISEASE CALLED

1340
00:59:28,746 --> 00:59:29,814
IDIOPATHIC INFLAMMATORY

1341
00:59:29,814 --> 00:59:30,582
MYOPATHIES.

1342
00:59:30,582 --> 00:59:32,584
I'LL TELL YOU ABOUT OTHER

1343
00:59:32,584 --> 00:59:38,056
EFFECTS MOLECULES PRESENT MAY

1344
00:59:38,056 --> 00:59:41,192
HAVE IN STROMAL CELLS.

1345
00:59:41,192 --> 00:59:44,596
AND SO AS ARTURO MENTIONED, THE

1346
00:59:44,596 --> 00:59:46,931
PROCESS OF NETs ONCE

1347
00:59:46,931 --> 00:59:47,999
NEUTROPHILS ENCOUNTER DANGEROUS

1348
00:59:47,999 --> 00:59:50,535
SIGNAL IS QUITE DRAMATIC AND

1349
00:59:50,535 --> 00:59:51,069
COMPLEX.

1350
00:59:51,069 --> 00:59:53,171
AND ONE OBSERVATION THAT HAS

1351
00:59:53,171 --> 00:59:55,773
BEEN MADE IS THAT DURING NET

1352
00:59:55,773 --> 00:59:57,508
FORMATION IN MANY CASES THERE'S

1353
00:59:57,508 --> 01:00:03,248
ACTIVATION OF A GROUP OF

1354
01:00:03,248 --> 01:00:05,650
ENZYMES, PADs, SPECIFICALLY

1355
01:00:05,650 --> 01:00:08,152
ONE OF THEM PAD 4 IS PRESENT

1356
01:00:08,152 --> 01:00:10,555
QUITE CHARACTERISTICALLY AND

1357
01:00:10,555 --> 01:00:11,689
SPECIFICALLY IN NEUTROPHILS.

1358
01:00:11,689 --> 01:00:19,063
AND WHAT THIS ENZYME DOES,

1359
01:00:19,063 --> 01:00:22,600
HISTONES PROMOTE THE ABILITY OF

1360
01:00:22,600 --> 01:00:24,135
CHROMATIN AMONG OTHER FACTORS

1361
01:00:24,135 --> 01:00:27,772
THAT ARTURO MENTIONED SUCH AS

1362
01:00:27,772 --> 01:00:29,741
NPO AND NEUTROPHIL ASSAYS, THIS

1363
01:00:29,741 --> 01:00:32,043
IS A KEY FACTOR IN THE FORMATION

1364
01:00:32,043 --> 01:00:36,848
OF NETs AND EXTRUSION INTO THE

1365
01:00:36,848 --> 01:00:37,282
EXTRACELLULAR SPACE.

1366
01:00:37,282 --> 01:00:39,450
GROUPS INCLUDING OURS HAVE BEEN

1367
01:00:39,450 --> 01:00:40,585
INTERESTED IN FINDING WHETHER

1368
01:00:40,585 --> 01:00:43,087
BLOCKING PAD ACTIVATION BECAUSE

1369
01:00:43,087 --> 01:00:45,356
IN MANY ANIMAL MODELS THEY HAVE

1370
01:00:45,356 --> 01:00:48,426
LOCATED PAD 4, EITHER WITH

1371
01:00:48,426 --> 01:00:50,895
CHEMICAL INHIBITORS OR THROUGH

1372
01:00:50,895 --> 01:00:52,297
GENETIC KNOCKOUTS WILL LIMIT

1373
01:00:52,297 --> 01:00:53,865
ABILITY OF NEUTROPHIL TO FORM

1374
01:00:53,865 --> 01:00:55,233
NETs AND EVEN IF NETs FORM

1375
01:00:55,233 --> 01:00:57,669
THE QUALITY OF THE NETs MAY BE

1376
01:00:57,669 --> 01:00:58,136
DIFFERENT.

1377
01:00:58,136 --> 01:01:00,305
AND SO JUST TO SHOW YOU THAT IN

1378
01:01:00,305 --> 01:01:02,840
A NUMBER OF ANIMAL MODELS WE

1379
01:01:02,840 --> 01:01:07,245
HAVE FOUND IF WE BLOCK NETs

1380
01:01:07,245 --> 01:01:08,980
CHEMICALLY, WE TRY CHEMICALLY

1381
01:01:08,980 --> 01:01:11,916
INHIBITORS OF PADs, WE'RE ABLE

1382
01:01:11,916 --> 01:01:15,420
TO INHIBIT ORGAN DAMAGE IN LUPUS

1383
01:01:15,420 --> 01:01:16,888
MODELS SEEING LESS RENAL

1384
01:01:16,888 --> 01:01:19,424
INFLAMMATION, LESS FORMATION OF

1385
01:01:19,424 --> 01:01:21,292
THE POSITION OF IMMUNE COMPLEX

1386
01:01:21,292 --> 01:01:23,661
IN THE KIDNEY, PROMINENT EFFECTS

1387
01:01:23,661 --> 01:01:24,462
ON IMPROVING VASCULAR FUNCTION

1388
01:01:24,462 --> 01:01:27,665
IN THE MICE.

1389
01:01:27,665 --> 01:01:32,437
WHEN WE BLOCK NETs THROUGH

1390
01:01:32,437 --> 01:01:33,738
BLOCKING PADS BLOOD VESSELS

1391
01:01:33,738 --> 01:01:36,274
DILATE BETTER, TIME TO CLOT UPON

1392
01:01:36,274 --> 01:01:37,775
STIMULATION IS PROLONGED MEANING

1393
01:01:37,775 --> 01:01:41,312
IT TAKES LONGER TO CLOT AND THIS

1394
01:01:41,312 --> 01:01:42,246
MAY PREVENT PATHOGENIC

1395
01:01:42,246 --> 01:01:42,547
THROMBOSIS.

1396
01:01:42,547 --> 01:01:47,418
THIS IS AN AVENUE THAT IS NOW

1397
01:01:47,418 --> 01:01:52,323
BEING EXPLORED AT THE

1398
01:01:52,323 --> 01:01:53,291
PRE-CLINICAL LEVEL BLOCKING

1399
01:01:53,291 --> 01:01:55,893
COULD HAVE THERAPEUTIC ROLES.

1400
01:01:55,893 --> 01:02:04,435
IN ANIMAL MODELS OF

1401
01:02:04,435 --> 01:02:06,237
ATHEROSCLEROSIS, APO-E PROTEIN,

1402
01:02:06,237 --> 01:02:15,013
IF WE BLOCK NET FORM  FORMATIONR

1403
01:02:15,013 --> 01:02:17,749
LIMIT PADs WE SEE LESS PLAQUE

1404
01:02:17,749 --> 01:02:19,183
FORMATION, HERE SIGNIFICANTLY

1405
01:02:19,183 --> 01:02:19,584
DEPRESSED.

1406
01:02:19,584 --> 01:02:23,788
AND THERE ARE LESS LIPIDS RICH

1407
01:02:23,788 --> 01:02:27,525
PLAQUE IN THE AORTA, IT TAKES

1408
01:02:27,525 --> 01:02:30,962
MICE LONGER TO CLOT UPON A

1409
01:02:30,962 --> 01:02:31,963
STIMULUS THAT LEADS TO CLOT

1410
01:02:31,963 --> 01:02:38,136
FORMATION, PROTECT IVE EFFECT IN

1411
01:02:38,136 --> 01:02:38,603
VASCULAR DAMAGE.

1412
01:02:38,603 --> 01:02:42,974
IF WE KNOCK OUT THE ENZYME WE

1413
01:02:42,974 --> 01:02:47,178
SEE SIGNIFICANT DECREASES IN

1414
01:02:47,178 --> 01:02:52,250
ATHEROSCLEROSIS IN THESE PRONE

1415
01:02:52,250 --> 01:02:55,453
MICE, JUST TO START SEE HOW

1416
01:02:55,453 --> 01:02:56,320
INHIBITING NETs MAY HAVE

1417
01:02:56,320 --> 01:02:58,089
THERAPEUTIC ROLES IN THESE

1418
01:02:58,089 --> 01:02:58,389
CONDITIONS.

1419
01:02:58,389 --> 01:03:06,364
I'LL  GO TO THE MOST PREVALENT,

1420
01:03:06,364 --> 01:03:10,435
1% OF THE POPULATION, SIMILAR TO

1421
01:03:10,435 --> 01:03:13,871
LUPUS THERE'S A CONTINUUM, AN

1422
01:03:13,871 --> 01:03:16,040
AREA OF PRE-CLINICAL IMMUNE

1423
01:03:16,040 --> 01:03:16,674
DYSREGULATION THAT INTERESTINGLY

1424
01:03:16,674 --> 01:03:18,576
MAY START AT THE LEVEL ALSO OF

1425
01:03:18,576 --> 01:03:21,612
MUCOSA IN THE LUNG AND OTHER

1426
01:03:21,612 --> 01:03:25,083
MUCOSAS, PEOPLE AT RISK TO

1427
01:03:25,083 --> 01:03:26,717
DEVELOP RHEUMATOID ARTHRITIS,

1428
01:03:26,717 --> 01:03:27,819
WORK BY THE UNIVERSITY OF

1429
01:03:27,819 --> 01:03:29,854
COLORADO AND WE WERE VERY LUCKY

1430
01:03:29,854 --> 01:03:31,856
TO COLLABORATE WITH HIM, WE

1431
01:03:31,856 --> 01:03:37,628
FOUND ACTUALLY IN LUNG MUCOSA

1432
01:03:37,628 --> 01:03:38,696
THERE'S SIGNIFICANT INCREASED

1433
01:03:38,696 --> 01:03:41,165
LEVELS OF NETs.

1434
01:03:41,165 --> 01:03:43,434
AND WHAT IS INTERESTING IS THAT

1435
01:03:43,434 --> 01:03:46,838
THESE NETs WHICH CARRY

1436
01:03:46,838 --> 01:03:57,215
INCREASED LEVELS OF ANTIGENS

1437
01:03:57,215 --> 01:04:00,017
BECAUSE PATHS ARE ACTIVATED MAY

1438
01:04:00,017 --> 01:04:05,022
EXPOSE IMMUNE SYSTEM, ENHANCED

1439
01:04:05,022 --> 01:04:08,192
EVEN BEFORE RHEUMATOID ARTHRITIS

1440
01:04:08,192 --> 01:04:09,494
BECOME EVIDENT.

1441
01:04:09,494 --> 01:04:13,764
AND KEY AUTOANTIBODY THAT IS

1442
01:04:13,764 --> 01:04:17,101
PRESENT IN RHEUMATOID ARTHRITIS,

1443
01:04:17,101 --> 01:04:21,172
PATHOGNOMONIC, ANTIBODIES THAT

1444
01:04:21,172 --> 01:04:22,440
TARGETS, USED IN DIAGNOSIS OF

1445
01:04:22,440 --> 01:04:22,807
DISEASE.

1446
01:04:22,807 --> 01:04:24,275
WE THINK EARLY ON DURING THE

1447
01:04:24,275 --> 01:04:26,110
DISEASE THERE MAY BE ENHANCED,

1448
01:04:26,110 --> 01:04:27,478
EVEN BEFORE DISEASE DEVELOPS,

1449
01:04:27,478 --> 01:04:29,180
NETs ARE HAPPENING IN THE

1450
01:04:29,180 --> 01:04:29,547
LUNGS.

1451
01:04:29,547 --> 01:04:33,584
AND THIS LEADS TO EXPANSION AND

1452
01:04:33,584 --> 01:04:35,186
DEVELOPMENT EVENTUALLY OF

1453
01:04:35,186 --> 01:04:36,087
RHEUMATOID ARTHRITIS THAT,

1454
01:04:36,087 --> 01:04:38,356
AGAIN, THE DISEASE PROGRESSES

1455
01:04:38,356 --> 01:04:40,458
CAN LEAD TO VERY SIGNIFICANT

1456
01:04:40,458 --> 01:04:43,427
DAMAGE AND DISABILITY.

1457
01:04:43,427 --> 01:04:49,500
AND SEVERAL YEARS AGO WE

1458
01:04:49,500 --> 01:04:50,434
HYPOTHESIZED ANTIGENS MAY BE

1459
01:04:50,434 --> 01:04:52,170
GENERATED, PERHAPS NETs WERE A

1460
01:04:52,170 --> 01:04:54,772
KEY INITIATOR IN THE DISEASE.

1461
01:04:54,772 --> 01:04:56,841
AND JUST TO SUMMARIZE THIS STORY

1462
01:04:56,841 --> 01:05:01,779
WHAT WE FOUND IS THAT THE

1463
01:05:01,779 --> 01:05:03,080
RHEUMATOID ARTHRITIS

1464
01:05:03,080 --> 01:05:03,481
MICROENVIRONMENT,

1465
01:05:03,481 --> 01:05:04,415
PRO-INFLAMMATORY, BOTH IN THE

1466
01:05:04,415 --> 01:05:07,385
SYSTEMIC BUT ALSO INSIDE THE

1467
01:05:07,385 --> 01:05:09,887
JOINTS WAS A CHARACTERIZED BY

1468
01:05:09,887 --> 01:05:11,422
ENHANCED FORMATION OF NETs

1469
01:05:11,422 --> 01:05:13,824
THAT MANY NEUTROPHILS ARE FOUND

1470
01:05:13,824 --> 01:05:15,726
IN THE SYNOVIAL FLUID OF

1471
01:05:15,726 --> 01:05:16,627
RHEUMATOID ARTHRITIS PATIENTS

1472
01:05:16,627 --> 01:05:18,396
AND THESE HAVE ENHANCED CAPACITY

1473
01:05:18,396 --> 01:05:23,801
TO FORM NETs, EXTERNALIZE,

1474
01:05:23,801 --> 01:05:26,871
REACH LEVELS OF ANTIGEN, LED TO

1475
01:05:26,871 --> 01:05:28,839
THE INDUCTION OF PHOTOANTIBODIES

1476
01:05:28,839 --> 01:05:31,409
THAT WERE DIRECTED AGAINST THE

1477
01:05:31,409 --> 01:05:33,144
PEPTIDE AND IN TURN

1478
01:05:33,144 --> 01:05:35,713
AUTOANTIBODIES CAN LEAD TO MORE

1479
01:05:35,713 --> 01:05:41,852
NET FORMATION CREATING A VICIOUS

1480
01:05:41,852 --> 01:05:43,054
CYCLE.

1481
01:05:43,054 --> 01:05:44,822
THESE AUTOANTIGENS HAD THE

1482
01:05:44,822 --> 01:05:48,326
ABILITY TO STIMULATE A KEY CELL

1483
01:05:48,326 --> 01:05:51,395
PRESENT IN THE RHEUMATOID -- IN

1484
01:05:51,395 --> 01:05:54,332
THE JOINT KNOWN TO BE IMPORTANT

1485
01:05:54,332 --> 01:05:59,437
IN THE PATHOGENESIS OF

1486
01:05:59,437 --> 01:06:02,974
RHEUMATOID ARTHRITIS,

1487
01:06:02,974 --> 01:06:04,775
SYNOVIAL -- FIBROBLAST IN

1488
01:06:04,775 --> 01:06:08,179
SYNOVIUM, AND THAT WHEN NETs

1489
01:06:08,179 --> 01:06:09,780
STIMULATED THE FIBROBLASTS THEY

1490
01:06:09,780 --> 01:06:11,315
BECAME PRO-INFLAMMATORY AND

1491
01:06:11,315 --> 01:06:11,782
AGGRESSIVE.

1492
01:06:11,782 --> 01:06:14,485
SO WE FOLLOW WITH THIS STORY AND

1493
01:06:14,485 --> 01:06:19,824
THIS LED US TO FIND THAT IN FACT

1494
01:06:19,824 --> 01:06:24,629
WHEN SYNOVIAL CYTO BLASTS

1495
01:06:24,629 --> 01:06:27,231
ENCOUNTER NETs, THEY HAVE THE

1496
01:06:27,231 --> 01:06:28,866
ABILITY TO INTERNALIZE, LEADING

1497
01:06:28,866 --> 01:06:31,435
TO ACTIVATION, AND THAT WHEN

1498
01:06:31,435 --> 01:06:33,437
THEY BECAME ACTIVATED THEY

1499
01:06:33,437 --> 01:06:38,175
ACQUIRE ANTIGEN PRESENTING CELL

1500
01:06:38,175 --> 01:06:39,210
CAPABILITIES, WENT TO ADAPTIVE

1501
01:06:39,210 --> 01:06:41,746
IMMUNE SYSTEM AND PRESENT

1502
01:06:41,746 --> 01:06:44,215
AUTOANTIGENS TO T CELLS WHICH

1503
01:06:44,215 --> 01:06:47,218
PROMOTE FORMATION OF THESE KEY

1504
01:06:47,218 --> 01:06:47,752
AUTOANTIBODIES PRESENT IN

1505
01:06:47,752 --> 01:06:52,023
RHEUMATOID ARTHRITIS, THE ONES

1506
01:06:52,023 --> 01:06:53,391
THAT TARGET SUGGESTING AGAIN

1507
01:06:53,391 --> 01:06:55,760
NETs MAY PLAY IMPORTANT ROLES

1508
01:06:55,760 --> 01:06:59,430
IN THE INITIATION.

1509
01:06:59,430 --> 01:07:03,301
NOT ONLY THAT WE KNOW THERE ARE

1510
01:07:03,301 --> 01:07:03,801
OTHER POST-TRANSLATIONAL

1511
01:07:03,801 --> 01:07:06,103
MODIFICATIONS PRESENT IN

1512
01:07:06,103 --> 01:07:08,005
RHEUMATOID ARTHRITIS, THE OTHER

1513
01:07:08,005 --> 01:07:18,382
ONE IS CARBAMYLATION.

1514
01:07:21,285 --> 01:07:24,755
AND WHAT WE FOUND YEARS AGO IS

1515
01:07:24,755 --> 01:07:26,691
THAT CARBAMYLATION PROTEINS ARE

1516
01:07:26,691 --> 01:07:29,060
ALSO PRESENT IN NETs, AND

1517
01:07:29,060 --> 01:07:32,396
SPECIFICALLY HISTONES CAN BE

1518
01:07:32,396 --> 01:07:35,499
FOUND IN NETs, AND IT IS

1519
01:07:35,499 --> 01:07:37,835
INTERESTING IN RHEUMATOID

1520
01:07:37,835 --> 01:07:42,773
ARTHRITIS PRESENCE TARGETING  TS

1521
01:07:42,773 --> 01:07:45,609
ASSOCIATED TO BONE DAMAGE.

1522
01:07:45,609 --> 01:07:47,378
PEOPLE THAT CARRY ANTIBODIES

1523
01:07:47,378 --> 01:07:50,114
HAVE ENHANCED RISK TO DEVELOP

1524
01:07:50,114 --> 01:07:52,350
BONE DAMAGE, ENHANCED BONE

1525
01:07:52,350 --> 01:07:55,453
RESORPTION, AND A FEW YEARS AGO

1526
01:07:55,453 --> 01:07:59,423
WE DESCRIBED POTENTIAL

1527
01:07:59,423 --> 01:08:02,259
MECHANISM, IN THAT NETs

1528
01:08:02,259 --> 01:08:05,429
CARRYING THIS HAVE ABILITY TO

1529
01:08:05,429 --> 01:08:07,431
ACTIVATE THROUGH ENGAGEMENT,

1530
01:08:07,431 --> 01:08:09,667
ALLOWING MONOCYTES TO UNDERGO

1531
01:08:09,667 --> 01:08:12,002
ACCELERATED PROGRAM WHERE THEY

1532
01:08:12,002 --> 01:08:13,771
BECOME ACTIVATED OSTEOBLASTS

1533
01:08:13,771 --> 01:08:15,439
THAT RESORB BONE AND ALSO

1534
01:08:15,439 --> 01:08:19,610
ACTIVATE T CELLS TO PRODUCE

1535
01:08:19,610 --> 01:08:23,481
OTHER PRO OSTEOGENIC OR BONE

1536
01:08:23,481 --> 01:08:26,550
ABSORPTION MOLECULES LEADING TO

1537
01:08:26,550 --> 01:08:28,486
NET FORMATION IMPORTANCE IN

1538
01:08:28,486 --> 01:08:30,688
INDUCING ARTHRITIS DEVELOPMENT

1539
01:08:30,688 --> 01:08:32,022
OR ACTIVATION OF SYNOVIAL

1540
01:08:32,022 --> 01:08:33,124
FIBROBLASTS BUT LEAD TO

1541
01:08:33,124 --> 01:08:35,459
END-ORGAN DAMAGE, TO BONE

1542
01:08:35,459 --> 01:08:36,026
DAMAGE.

1543
01:08:36,026 --> 01:08:38,996
AND FINALLY MY THIRD EXAMPLE OF

1544
01:08:38,996 --> 01:08:41,165
AUTOIMMUNITY DEVELOPMENT LINKED

1545
01:08:41,165 --> 01:08:46,937
TO NETs IS INDUCED MODEL OF

1546
01:08:46,937 --> 01:08:49,340
AUTOIMMUNITY WHICH IS

1547
01:08:49,340 --> 01:08:51,876
VASCULITIS, ANOTHER FASCINATING

1548
01:08:51,876 --> 01:08:56,881
CONDITION WHERE LEVAMISOLE IS

1549
01:08:56,881 --> 01:08:59,817
UNIT IN MEDICINE, AN AGONIST,

1550
01:08:59,817 --> 01:09:03,020
MOST COCAINE CONSUMED IN THE

1551
01:09:03,020 --> 01:09:04,221
U.S. IS CONTAMINATED WITH

1552
01:09:04,221 --> 01:09:07,425
LEVAMISOLE, AND THERE ARE CASES

1553
01:09:07,425 --> 01:09:10,161
ASSOCIATED WITH PATIENTS THAT

1554
01:09:10,161 --> 01:09:12,696
USE COCAINE AND DEVELOP

1555
01:09:12,696 --> 01:09:13,697
CHARACTERISTICS TYPE OF

1556
01:09:13,697 --> 01:09:16,600
VASCULITIS THAT WAS INDUCED BY

1557
01:09:16,600 --> 01:09:16,934
CONTAMINATION.

1558
01:09:16,934 --> 01:09:25,309
TYPICALLY THESE PATIENTS DEVELOP

1559
01:09:25,309 --> 01:09:27,044
FEVER, PURPURIC RASHES, IN THE

1560
01:09:27,044 --> 01:09:31,549
NOSE, EXTREMITIES, A MIX OF

1561
01:09:31,549 --> 01:09:32,416
ANTIBODIES TARGET AUTOANTIGENS,

1562
01:09:32,416 --> 01:09:35,252
PRESENT IN NETs.

1563
01:09:35,252 --> 01:09:39,557
MANY DEVELOP LOW LEVELS OF

1564
01:09:39,557 --> 01:09:40,825
NEUTROFIRST SUGGESTING

1565
01:09:40,825 --> 01:09:42,026
NEUTROPHILS MAY BE DYING.

1566
01:09:42,026 --> 01:09:46,330
WHEN THE PATIENT STOP THE

1567
01:09:46,330 --> 01:09:48,299
COCAINE AND EXPOSURE TO

1568
01:09:48,299 --> 01:09:49,700
LEVAMISOLE THIS VASCULITIS

1569
01:09:49,700 --> 01:09:50,134
RESULTS.

1570
01:09:50,134 --> 01:09:53,070
IN COLLABORATION WE FOUND A

1571
01:09:53,070 --> 01:09:54,872
PATIENT THAT HAD DEVELOPED

1572
01:09:54,872 --> 01:09:55,739
LEVAMISOLE INDUCED VASCULITIS,

1573
01:09:55,739 --> 01:09:57,708
FOUND THIS PATIENT HAD

1574
01:09:57,708 --> 01:10:00,144
SIGNIFICANT LEVELS OF LOW

1575
01:10:00,144 --> 01:10:02,079
DENSITY GRANULOCYTES FORMING

1576
01:10:02,079 --> 01:10:03,414
PROFOUND LEVELS OF NETs, AND

1577
01:10:03,414 --> 01:10:05,816
WE ALSO HAD ABILITY TO STUDY

1578
01:10:05,816 --> 01:10:08,252
TISSUES OF PATIENTS WITH

1579
01:10:08,252 --> 01:10:10,421
LEVAMISOLE INDUCED VASCULITIS,

1580
01:10:10,421 --> 01:10:13,491
TISSUES WERE INFILTRATED BY

1581
01:10:13,491 --> 01:10:13,924
NETTING NEUTROPHILS.

1582
01:10:13,924 --> 01:10:18,863
WHAT WE FOUND WAS A LINK BETWEEN

1583
01:10:18,863 --> 01:10:20,798
NEURAL IMMUNOMODULATION IN THAT

1584
01:10:20,798 --> 01:10:24,602
AS I MENTIONED LEVAMISOLE

1585
01:10:24,602 --> 01:10:27,671
STIMULATES RECEPTORS, USED WITH

1586
01:10:27,671 --> 01:10:29,707
MEDICINE, SO WE ASKED HERE IN

1587
01:10:29,707 --> 01:10:32,243
VITRO YOU SEE WHAT HAPPENS IF WE

1588
01:10:32,243 --> 01:10:34,712
ADD LEVAMISOLE WE SEE PROFOUND

1589
01:10:34,712 --> 01:10:36,680
INDUCTION OF NETs.

1590
01:10:36,680 --> 01:10:39,416
AND WE HYPOTHESIZE PERHAPS IT

1591
01:10:39,416 --> 01:10:41,919
WAS THROUGH INDUCTION OF

1592
01:10:41,919 --> 01:10:43,554
RECEPTORS NETs WERE BEING

1593
01:10:43,554 --> 01:10:43,921
INDUCED.

1594
01:10:43,921 --> 01:10:54,431
IF YOU ADD ANOTHER MUSCARINIC

1595
01:10:55,199 --> 01:10:56,867
ACID, INDUCED NETs.

1596
01:10:56,867 --> 01:10:59,937
AND ATROPINE, WE WERE ABLE TO

1597
01:10:59,937 --> 01:11:01,572
BLOCK ABILITY OF LEVAMISOLE TO

1598
01:11:01,572 --> 01:11:02,606
INDUCE NETs.

1599
01:11:02,606 --> 01:11:04,375
QUANTITY VERSUS QUALITY WE FOUND

1600
01:11:04,375 --> 01:11:06,210
IF WE INDUCE NETs WITH

1601
01:11:06,210 --> 01:11:08,145
LEVAMISOLE, THEY HAVE AN

1602
01:11:08,145 --> 01:11:09,480
ENHANCED ABILITY TO DAMAGE BLOOD

1603
01:11:09,480 --> 01:11:11,048
VESSELS WHEN COMPARED TO NETs

1604
01:11:11,048 --> 01:11:13,551
INDUCED BY OTHER TYPES OF

1605
01:11:13,551 --> 01:11:14,485
STIMULATION, SUGGESTING AGAIN

1606
01:11:14,485 --> 01:11:16,453
HOW NETs ARE INDUCED MAY BE

1607
01:11:16,453 --> 01:11:20,291
KEY IN WHETHER THEY ARE

1608
01:11:20,291 --> 01:11:22,893
PRO-INFLAMMATORY AND HARM

1609
01:11:22,893 --> 01:11:23,193
VASCULATURE.

1610
01:11:23,193 --> 01:11:25,729
AND SO JUST IN MY LAST MINUTES

1611
01:11:25,729 --> 01:11:28,265
HOW CAN WE TARGET NET FORMATION?

1612
01:11:28,265 --> 01:11:33,370
COULD THIS BE POTENTIALLY USED

1613
01:11:33,370 --> 01:11:35,806
TO MITIGATE AUTOIMMUNITY AND

1614
01:11:35,806 --> 01:11:38,275
VASCULAR DAMAGE?

1615
01:11:38,275 --> 01:11:39,944
INHIBITION IS BEING

1616
01:11:39,944 --> 01:11:42,880
INVESTIGATED, MODULATING ROS

1617
01:11:42,880 --> 01:11:44,315
PRODUCTION IS BEING INVESTIGATED

1618
01:11:44,315 --> 01:11:48,686
AS A WAY TO TARGET RESPONSE OF

1619
01:11:48,686 --> 01:11:50,554
NEUTROPHILS TO STIMULUS THAT

1620
01:11:50,554 --> 01:11:53,290
COULD DAMPEN NET FORMATION AND

1621
01:11:53,290 --> 01:11:55,092
RECENTLY FOCUSED ON A -- THIS IS

1622
01:11:55,092 --> 01:11:56,694
JUST TO ALSO MENTION THERE ARE

1623
01:11:56,694 --> 01:11:58,462
DRUGS THAT WE'RE ALREADY USING

1624
01:11:58,462 --> 01:12:02,032
AND HAVE USED FOR MANY YEARS TO

1625
01:12:02,032 --> 01:12:04,168
TREAT AUTOIMMUNITY THAT HAVE

1626
01:12:04,168 --> 01:12:07,071
BEEN UNCLEAR WHAT MECHANISM OF

1627
01:12:07,071 --> 01:12:08,105
ACTION IS, ONE IS

1628
01:12:08,105 --> 01:12:08,939
ANTI-MALARIALS, QUITE BENEFICIAL

1629
01:12:08,939 --> 01:12:10,941
FOR MANY LUPUS PATIENTS AND

1630
01:12:10,941 --> 01:12:18,082
PATIENTS WITH OTHER AUTOIMMUNE

1631
01:12:18,082 --> 01:12:21,819
DISEASES, THEY CAN INHIBIT NET

1632
01:12:21,819 --> 01:12:25,522
FORMATION AND EXTRUSION OF

1633
01:12:25,522 --> 01:12:27,424
NETs AND CYTOTOXIC MOLECULES

1634
01:12:27,424 --> 01:12:29,760
PRESENT BUT ALSO ABILITY OF

1635
01:12:29,760 --> 01:12:31,428
NETs TO BE INTERNALIZED BY

1636
01:12:31,428 --> 01:12:35,299
OTHER CELLS CAN BE INHIBITED

1637
01:12:35,299 --> 01:12:37,201
WITH ANTI-MALARIALS, DRUGS FOR

1638
01:12:37,201 --> 01:12:41,305
MANY YEARS WOULD THE KNOWING HOW

1639
01:12:41,305 --> 01:12:44,274
THEY WORK IN PART, IN PART

1640
01:12:44,274 --> 01:12:46,677
INHIBIT NET FORMATION.

1641
01:12:46,677 --> 01:12:49,146
THE OTHER WAY BY WHICH WE'VE

1642
01:12:49,146 --> 01:12:50,280
FOCUSED ON WHETHER INHIBITING

1643
01:12:50,280 --> 01:12:52,316
NETs MAY BE BENEFICIAL IS CAN

1644
01:12:52,316 --> 01:12:55,052
WE INHIBIT THE ABILITY OF NETs

1645
01:12:55,052 --> 01:12:55,853
TO HARM THE VASCULATURE BECAUSE

1646
01:12:55,853 --> 01:12:59,957
AT THE END WE THINK THIS IS A

1647
01:12:59,957 --> 01:13:01,825
KEY MECHANISM BY WHICH NETs

1648
01:13:01,825 --> 01:13:05,262
ARE DELETERIOUS IN AUTOIMMUNITY.

1649
01:13:05,262 --> 01:13:08,098
A KEY PATHWAY CYTOKINE SIGNAL

1650
01:13:08,098 --> 01:13:10,901
MAY BE CONDUCIVE TO NET

1651
01:13:10,901 --> 01:13:14,004
FORMATION IS JAK/STAT PATHWAY,

1652
01:13:14,004 --> 01:13:14,872
ACTIVE IN NEUTROPHILS, WE FOUND

1653
01:13:14,872 --> 01:13:16,607
WHEN TRIGGERED MAY LEAD TO

1654
01:13:16,607 --> 01:13:18,575
INDUCTION OF NETs.

1655
01:13:18,575 --> 01:13:20,644
IN FACT, IN CLINICAL TRIALS IN

1656
01:13:20,644 --> 01:13:23,180
LUPUS PATIENTS WE FOUND THAT

1657
01:13:23,180 --> 01:13:32,856
WHEN THE PATIENTS TOOK THE JAK

1658
01:13:32,856 --> 01:13:34,725
INHIBITORS COMPARED TO PLACEBO,

1659
01:13:34,725 --> 01:13:36,126
ASSOCIATED WITH DECREASED LEVELS

1660
01:13:36,126 --> 01:13:37,561
OF NET FORMATION IN THESE

1661
01:13:37,561 --> 01:13:38,295
PATIENTS EVEN DURING SHORT

1662
01:13:38,295 --> 01:13:41,265
PERIOD OF TIME THEY TOOK THE

1663
01:13:41,265 --> 01:13:41,832
DRUG.

1664
01:13:41,832 --> 01:13:43,934
AND TRANSLATED INTO FINDINGS NOT

1665
01:13:43,934 --> 01:13:45,402
ONLY IMMUNE DYSREGULATION WAS

1666
01:13:45,402 --> 01:13:47,571
IMPROVED BUT THAT LEVEL OF

1667
01:13:47,571 --> 01:13:48,539
VASCULAR DAMAGE OR VASCULAR

1668
01:13:48,539 --> 01:13:50,808
DYSFUNCTION THAT WE FOUND IN THE

1669
01:13:50,808 --> 01:13:57,147
LUPUS PATIENTS WENT DOWN WHEN WE

1670
01:13:57,147 --> 01:13:59,450
DECREASED -- INHIBITS JAK AND

1671
01:13:59,450 --> 01:14:00,317
LOW DENSITY NETs SUGGESTING

1672
01:14:00,317 --> 01:14:03,187
THERE MAY BE WAYS TO REGULATE

1673
01:14:03,187 --> 01:14:04,221
NET FORMATION THAT MAY BE

1674
01:14:04,221 --> 01:14:06,690
BENEFICIAL TO PATIENTS IN THE

1675
01:14:06,690 --> 01:14:07,157
FUTURE.

1676
01:14:07,157 --> 01:14:09,293
SO I WILL END HERE JUST TO

1677
01:14:09,293 --> 01:14:11,528
SUMMARIZE SOME KEY ASPECTS OF MY

1678
01:14:11,528 --> 01:14:13,030
TALK.

1679
01:14:13,030 --> 01:14:15,432
ONE IS THAT SEX DIFFERENCES IN

1680
01:14:15,432 --> 01:14:16,366
NEUTROPHIL BIOLOGY MAY

1681
01:14:16,366 --> 01:14:19,169
CONTRIBUTE TO AUTOIMMUNITY AND

1682
01:14:19,169 --> 01:14:20,504
FOR INFECTIONS THAT WE BELIEVE

1683
01:14:20,504 --> 01:14:22,039
NETs WHEN PROCESS GOES AWRY

1684
01:14:22,039 --> 01:14:26,310
MAY SERVE AS IMPORTANT SOURCE OF

1685
01:14:26,310 --> 01:14:34,585
MODIFIED AUTOANTIGENS, NUCLEIC

1686
01:14:34,585 --> 01:14:36,353
ACID AND, AND CHARACTERIZING

1687
01:14:36,353 --> 01:14:41,058
NEUTROPHIL SUBSETS, NOT ALL, AND

1688
01:14:41,058 --> 01:14:44,261
ABILITY IN NET FORMATION MAY

1689
01:14:44,261 --> 01:14:45,462
CONTRIBUTE TO MITIGATE END-ORGAN

1690
01:14:45,462 --> 01:14:51,435
DAMAGE IN LUPUS AND OTHER

1691
01:14:51,435 --> 01:14:52,936
AUTOIMMUNE DISEASE.

1692
01:14:52,936 --> 01:14:54,872
I MENTION MANY FORMER AROUND

1693
01:14:54,872 --> 01:14:57,274
CURRENT MEMBERS OF NIGH OF MY

1694
01:14:57,274 --> 01:14:58,842
LAB, COLLABORATORS AND GREAT

1695
01:14:58,842 --> 01:15:00,778
SUPPORT FROM NIAMS AND THE NIH

1696
01:15:00,778 --> 01:15:01,211
IN GENERAL.

1697
01:15:01,211 --> 01:15:06,917
THANK YOU VERY MUCH FOR YOUR

1698
01:15:06,917 --> 01:15:07,584
ATTENTION.

1699
01:15:07,584 --> 01:15:09,853
>> WELL, THANK YOU VERY
MUCH,

1700
01:15:09,853 --> 01:15:14,224
MARIANA AND ARTURO, FOR THIS

1701
01:15:14,224 --> 01:15:15,159
FASCINATING TOUR DE FORCE

1702
01:15:15,159 --> 01:15:19,797
TELLING US ABOUT BASIC SCIENCE

1703
01:15:19,797 --> 01:15:23,433
AND TRANSLATIONAL ASPECTS OF NET

1704
01:15:23,433 --> 01:15:24,568
OSIS.

1705
01:15:24,568 --> 01:15:26,703
WE HAVE AN ENORMOUS NUMBER OF

1706
01:15:26,703 --> 01:15:27,538
QUESTIONS BEFORE US.

1707
01:15:27,538 --> 01:15:29,506
NOT THAT THAT'S INDICATING IN A

1708
01:15:29,506 --> 01:15:32,309
THERE'S ANY LACK OF CLARITY OF

1709
01:15:32,309 --> 01:15:35,112
THE PRESENTATIONS BUT RATHER

1710
01:15:35,112 --> 01:15:36,580
EXCITEMENT ABOUT THE IDEAS THAT

1711
01:15:36,580 --> 01:15:38,615
HAVE BEEN PUT FORTH.

1712
01:15:38,615 --> 01:15:42,186
SO, WE'LL START OUT WITH ONE

1713
01:15:42,186 --> 01:15:44,321
FROM OUR AUDIENCE ASKING FOR A

1714
01:15:44,321 --> 01:15:47,124
LITTLE BIT MORE DETAIL IN TERMS

1715
01:15:47,124 --> 01:15:51,161
OF HOW HISTONES HAVE THEIR

1716
01:15:51,161 --> 01:15:52,429
ANTIMICROBIAL EFFECT AND PERHAPS

1717
01:15:52,429 --> 01:15:56,733
WE CAN START WITH ARTURO, WITH

1718
01:15:56,733 --> 01:15:57,901
THAT, BUT MARIANA MAY HAVE

1719
01:15:57,901 --> 01:16:02,439
SOMETHING TO ADD AS WELL.

1720
01:16:02,439 --> 01:16:02,673
ARTURO?

1721
01:16:02,673 --> 01:16:07,411
>>  WE DON'T KNOW, THE
SHORT

1722
01:16:07,411 --> 01:16:07,678
ANSWER.

1723
01:16:07,678 --> 01:16:08,378
HISTONES ARE CATIONIC, THAT

1724
01:16:08,378 --> 01:16:10,614
MIGHT BE ONE OF THE WAYS, BUT IT

1725
01:16:10,614 --> 01:16:13,150
IS NOT THE ONLY THING.

1726
01:16:13,150 --> 01:16:14,318
THEY ARE EQUALLY CATIONIC BUT

1727
01:16:14,318 --> 01:16:19,256
NOT AS STRONG AS ANTIMICROBIALS.

1728
01:16:19,256 --> 01:16:21,925
HISTONE H-1 WE KNOW THAT IN

1729
01:16:21,925 --> 01:16:25,295
STAPH AUREUS IT IS RECRUITED

1730
01:16:25,295 --> 01:16:29,233
INTO THE SURFACE OF THE CELL,

1731
01:16:29,233 --> 01:16:31,301
THEN BREAKS THE FORCE BETWEEN

1732
01:16:31,301 --> 01:16:32,236
THE MEMBRANES.

1733
01:16:32,236 --> 01:16:36,340
AND WE'VE SEEN BINDING EVER H1

1734
01:16:36,340 --> 01:16:37,741
TO PROKARYOTIC DNA BUT DON'T

1735
01:16:37,741 --> 01:16:40,043
KNOW IF THAT'S SIGNIFICANT.

1736
01:16:40,043 --> 01:16:42,779
>> SO, THAT'S REALLY

1737
01:16:42,779 --> 01:16:43,447
FASCINATING.

1738
01:16:43,447 --> 01:16:45,716
AND YOU MENTIONED AT LEAST

1739
01:16:45,716 --> 01:16:48,986
HYPOTHESIZED THAT THERE COULD BE

1740
01:16:48,986 --> 01:16:53,056
SOME SORT OF EVOLUTIONARY THING

1741
01:16:53,056 --> 01:16:57,527
GOING ON HERE, WHERE EUKARYOTES

1742
01:16:57,527 --> 01:16:59,863
BASICALLY HAVE EVOLVED HISTONES

1743
01:16:59,863 --> 01:17:02,065
AT ABOUT THE TIME THAT THEY

1744
01:17:02,065 --> 01:17:04,935
DEVELOPED NUCLEUS AND AT ABOUT

1745
01:17:04,935 --> 01:17:07,170
THE TIME THAT BACTERIA WOULD BE

1746
01:17:07,170 --> 01:17:09,006
A THREAT TO THEM.

1747
01:17:09,006 --> 01:17:15,379
DO CANDIDA YEASTS HAVE HISTONES?

1748
01:17:15,379 --> 01:17:16,914
>> YES.

1749
01:17:16,914 --> 01:17:19,149
ALL EUKARYOTES HAVE HISTONES,

1750
01:17:19,149 --> 01:17:19,650
THEY ARE EXTRAORDINARILY

1751
01:17:19,650 --> 01:17:26,423
CONSERVED THROUGHOUT  EUKARYOTIC

1752
01:17:26,423 --> 01:17:26,690
EVOLUTION.

1753
01:17:26,690 --> 01:17:29,059
HISTONE-LIKE PROTEINS ARE MUCH

1754
01:17:29,059 --> 01:17:31,228
MORE SIMILAR TO EUKARYOTIC

1755
01:17:31,228 --> 01:17:35,299
HISTONES THAN PROTEINS THAT BIND

1756
01:17:35,299 --> 01:17:36,366
TO PROKARYOTIC DNA.

1757
01:17:36,366 --> 01:17:38,602
GREAT TO RECRUIT SOMEBODY

1758
01:17:38,602 --> 01:17:42,172
WORKING TO SEE WHAT IS THE

1759
01:17:42,172 --> 01:17:43,507
FUNCTION OF HISTONES.

1760
01:17:43,507 --> 01:17:49,446
>> YEAH, THAT WOULD BE

1761
01:17:49,446 --> 01:17:49,880
FASCINATING.

1762
01:17:49,880 --> 01:17:51,281
NOW PERHAPS A QUESTION FOR

1763
01:17:51,281 --> 01:17:53,917
MARIANA WITH REGARD TO LOW

1764
01:17:53,917 --> 01:17:56,219
DENSITY NEUTROPHILS, AND THEIR

1765
01:17:56,219 --> 01:17:59,189
PROCLIVITY TO PRODUCE NETs.

1766
01:17:59,189 --> 01:18:01,825
AND SO THE QUESTION HAS TO DO

1767
01:18:01,825 --> 01:18:04,161
WITH THE ISSUE OF WHETHER

1768
01:18:04,161 --> 01:18:09,666
NETOSIS IS SORT OF A FATAL

1769
01:18:09,666 --> 01:18:10,934
SUICIDAL ACTION BY NEUTROPHILS,

1770
01:18:10,934 --> 01:18:12,903
WHETHER THESE LOW DENSITY

1771
01:18:12,903 --> 01:18:14,071
NEUTROPHILS, WHETHER THEY ARE

1772
01:18:14,071 --> 01:18:17,240
PRECURSORS EVER CELLS THAT GO ON

1773
01:18:17,240 --> 01:18:19,242
TO MAKE NETs, YOU'RE CATCHING

1774
01:18:19,242 --> 01:18:21,979
THEM IN THE ACT OF, YOU KNOW,

1775
01:18:21,979 --> 01:18:23,180
MAKING NETs IN THE

1776
01:18:23,180 --> 01:18:24,514
CIRCULATION, WHAT'S GOING ON

1777
01:18:24,514 --> 01:18:26,616
WITH THOSE LOW DENSITY

1778
01:18:26,616 --> 01:18:27,050
NEUTROPHILS?

1779
01:18:27,050 --> 01:18:28,685
>>  YEAH, SO GREAT
QUESTION.

1780
01:18:28,685 --> 01:18:30,554
AND WE DON'T HAVE A FULL ANSWER.

1781
01:18:30,554 --> 01:18:32,956
WE'RE SPENDING A LOT OF TIME

1782
01:18:32,956 --> 01:18:34,591
TRYING TO FIGURE OUT.

1783
01:18:34,591 --> 01:18:38,428
WE KNOW IF YOU LOOK

1784
01:18:38,428 --> 01:18:39,429
TRANSCRIPTIONALLY AT PROTEOMIC

1785
01:18:39,429 --> 01:18:42,599
LEVEL THEY ARE DIFFERENT FROM

1786
01:18:42,599 --> 01:18:43,667
CELLS NORMALLY THEY EXTEND FROM

1787
01:18:43,667 --> 01:18:47,437
THE SAME PATIENT BUT ALSO FROM

1788
01:18:47,437 --> 01:18:48,438
HEALTHY VOLUNTEERS.

1789
01:18:48,438 --> 01:18:52,476
AND THEY LOOK SLIGHTLY LESS

1790
01:18:52,476 --> 01:18:56,580
MATURE BUT STILL HAVE THE FULL

1791
01:18:56,580 --> 01:18:57,481
MACHINERY OF MATURE NEUTROPHIL,

1792
01:18:57,481 --> 01:19:01,418
SO I WOULD SAY MAJORITY OF THESE

1793
01:19:01,418 --> 01:19:02,753
CELLS ARE MATURE.

1794
01:19:02,753 --> 01:19:05,389
SOME HETEROGENEITY, SOME APPEAR

1795
01:19:05,389 --> 01:19:07,424
TO BE LESS MATURE, BUT

1796
01:19:07,424 --> 01:19:10,293
ORIGINALLY THE CELLS WERE FULLY

1797
01:19:10,293 --> 01:19:11,495
A MATURE PHENOTYPE, WE HAVEN'T

1798
01:19:11,495 --> 01:19:15,832
BEEN ABLE TO RECAPITULATE, THAT

1799
01:19:15,832 --> 01:19:17,934
GOES WITH ENHANCED ABILITY TO

1800
01:19:17,934 --> 01:19:20,604
FORM NETs, TYPICALLY HAVE TO

1801
01:19:20,604 --> 01:19:23,106
BE MATURE TO FORM.

1802
01:19:23,106 --> 01:19:25,542
WHERE DO THEY COME FROM, ARE

1803
01:19:25,542 --> 01:19:26,610
THEY DIFFERENT FROM GET-GO IN

1804
01:19:26,610 --> 01:19:28,345
BONE MARROW OR SHAPED BY

1805
01:19:28,345 --> 01:19:30,580
ENCOUNTER IN VARIOUS ORGANS OR

1806
01:19:30,580 --> 01:19:33,316
AS THEY GO THROUGH CIRCULATION

1807
01:19:33,316 --> 01:19:36,553
DO THEY HAVE ENHANCED LIFE, MORE

1808
01:19:36,553 --> 01:19:39,022
SENESCENT, WE'VE BEEN TRYING TO

1809
01:19:39,022 --> 01:19:40,123
ADDRESS BUT PROVING TO TAKE

1810
01:19:40,123 --> 01:19:42,926
LONGER THAN WE WERE HOPING.

1811
01:19:42,926 --> 01:19:45,262
ONE ISSUE THAT WE HAVEN'T FOUND

1812
01:19:45,262 --> 01:19:47,197
A MOUSE COUNTERPART, SO THAT

1813
01:19:47,197 --> 01:19:48,765
MAKES IT HARDER.

1814
01:19:48,765 --> 01:19:50,067
MOUSE NEUTROPHILS ARE REALLY

1815
01:19:50,067 --> 01:19:51,268
QUITE DIFFERENT FROM HUMAN

1816
01:19:51,268 --> 01:19:55,439
NEUTROPHILS SO WE HAVE TO DO THE

1817
01:19:55,439 --> 01:19:57,140
STUDIES IN HUMANS, SO THAT'S

1818
01:19:57,140 --> 01:19:58,775
WHAT'S BEEN FASCINATING BUT

1819
01:19:58,775 --> 01:20:00,544
TAKING LONGER THAN WE HOPED TO

1820
01:20:00,544 --> 01:20:02,746
IDENTIFY WHAT THE CELLS ARE.

1821
01:20:02,746 --> 01:20:04,347
>> WELL, IT SHOWS THERE'S
REALLY

1822
01:20:04,347 --> 01:20:08,785
SOME SUBTLETY TO THE WHOLE

1823
01:20:08,785 --> 01:20:08,985
THING.

1824
01:20:08,985 --> 01:20:11,154
DO HEALTHY INDIVIDUALS HAVE

1825
01:20:11,154 --> 01:20:13,924
LOWER NUMBERS PERHAPS OF THESE

1826
01:20:13,924 --> 01:20:14,491
LOW DENSITY NEUTROPHILS?

1827
01:20:14,491 --> 01:20:15,158
>>  THEY DO.

1828
01:20:15,158 --> 01:20:17,194
SO WHAT WE HAVE FOUND IS THAT

1829
01:20:17,194 --> 01:20:20,430
THEY HAVE VERY SMALL AMOUNTS, SO

1830
01:20:20,430 --> 01:20:21,898
IT'S SOMETHING THAT EVERY HUMAN

1831
01:20:21,898 --> 01:20:25,202
APPEARS TO HAVE, BUT CELLS

1832
01:20:25,202 --> 01:20:27,070
BECOME MUCH MORE EXPANDED IN

1833
01:20:27,070 --> 01:20:30,474
CERTAIN DISEASES SUCH AS

1834
01:20:30,474 --> 01:20:30,974
AUTOIMMUNE DISEASES.

1835
01:20:30,974 --> 01:20:32,175
THEY WERE DESCRIBING COVID, BUT

1836
01:20:32,175 --> 01:20:35,445
IN COVID WE FOUND IT TO BE

1837
01:20:35,445 --> 01:20:40,117
DIFFERENT PHENOTYPICALLY SO THEY

1838
01:20:40,117 --> 01:20:41,384
WERE MORE SUPPRESSOR PHENOTYPE

1839
01:20:41,384 --> 01:20:47,157
SITES THAN WHAT WE SAW WITH

1840
01:20:47,157 --> 01:20:47,624
AUTOIMMUNITY, NOT SEE

1841
01:20:47,624 --> 01:20:49,826
SUPPRESSION, IF ANYTHING SEEING

1842
01:20:49,826 --> 01:20:51,461
MORE INFLAMMATION.

1843
01:20:51,461 --> 01:20:52,562
>> I'LL BITE MY TONGUE AND

1844
01:20:52,562 --> 01:20:58,368
BROKEN THIS UP BUT YOU DID

1845
01:20:58,368 --> 01:21:00,003
MENTION HYDROXYCHLOROQUINE,

1846
01:21:00,003 --> 01:21:01,138
NETs, NOW COVID,

1847
01:21:01,138 --> 01:21:02,239
NET-FORMING -- WELL, ANYWAY, I'M

1848
01:21:02,239 --> 01:21:06,042
NOT GOING ANY FURTHER WITH THAT.

1849
01:21:06,042 --> 01:21:09,613
BUT IN ANY CASE THERE'S ANOTHER

1850
01:21:09,613 --> 01:21:11,648
DETAIL ORIENTED MEMBER OF OUR

1851
01:21:11,648 --> 01:21:14,217
AUDIENCE WHO WANTS TO KNOW

1852
01:21:14,217 --> 01:21:17,988
WHETHER BASOPHILS AND

1853
01:21:17,988 --> 01:21:20,457
EOSINOPHILS CAN UNDERGO NET-OSIS

1854
01:21:20,457 --> 01:21:26,463
OR JUST NEUTROPHILS THAT DO?

1855
01:21:26,463 --> 01:21:33,170
>>  THEY CAN, ALL
GRANULOCYTES

1856
01:21:33,170 --> 01:21:35,438
CAN MAKE NETs.

1857
01:21:35,438 --> 01:21:35,705
>> OKAY.

1858
01:21:35,705 --> 01:21:36,773
>> THEY BOTH CAN.

1859
01:21:36,773 --> 01:21:37,774
>> UH-HUH, OKAY.

1860
01:21:37,774 --> 01:21:40,744
ALL RIGHT.

1861
01:21:40,744 --> 01:21:45,749
SO THEN HERE'S ANOTHER QUESTION.

1862
01:21:45,749 --> 01:21:48,251
THE AUDIENCE MEMBER WANTS TO

1863
01:21:48,251 --> 01:21:49,819
KNOW CAN CGD PATIENTS THAT DO

1864
01:21:49,819 --> 01:21:55,759
NOT MAKE NETs DEVELOP

1865
01:21:55,759 --> 01:21:56,193
AUTOIMMUNE DISEASES?

1866
01:21:56,193 --> 01:22:00,363
>>  YES. 

1867
01:22:00,363 --> 01:22:00,830
>> YES, THEY CAN --

1868
01:22:00,830 --> 01:22:03,466
>> I THINK THIS IS AN

1869
01:22:03,466 --> 01:22:04,834
INTERESTING QUESTION.

1870
01:22:04,834 --> 01:22:09,439
MOTHERS OF CGD KIDS ARE MORE

1871
01:22:09,439 --> 01:22:10,874
PRONE TO AUTOIMMUNITY, IS THAT

1872
01:22:10,874 --> 01:22:11,474
TRUE?

1873
01:22:11,474 --> 01:22:12,842
>> YES, WE'VE SEEN
PATIENTS THAT

1874
01:22:12,842 --> 01:22:16,746
DEVELOP A VARIETY OF AUTOIMMUNE

1875
01:22:16,746 --> 01:22:18,949
INFLAMMATORY, LUPUS, COLITIS, SO

1876
01:22:18,949 --> 01:22:22,319
IT'S NOT UNCOMMON TO SEE

1877
01:22:22,319 --> 01:22:23,220
AUTOIMMUNITY DEVELOPING.

1878
01:22:23,220 --> 01:22:27,424
WHAT I MAY SAY IN OUR HANDS

1879
01:22:27,424 --> 01:22:30,193
WE'VE FOUND PRESENCE IN SOME OF

1880
01:22:30,193 --> 01:22:33,697
THESE PATIENTS THAT HAVE

1881
01:22:33,697 --> 01:22:38,468
COEXISTING CGD AND AUTOIMMUNITY,

1882
01:22:38,468 --> 01:22:41,071
THEY MAKE EVEN A -- OXIDASE IS

1883
01:22:41,071 --> 01:22:43,707
IMPAIRED, SEEM TO HAVE AN

1884
01:22:43,707 --> 01:22:47,978
ABILITY TO SYNTHESIZE

1885
01:22:47,978 --> 01:22:49,279
MITOCHONDRIAL ROS, AND WE'VE

1886
01:22:49,279 --> 01:22:51,448
HYPOTHESIZED THIS COULD BE

1887
01:22:51,448 --> 01:22:54,484
ALTERNATIVE MECHANISM THAT

1888
01:22:54,484 --> 01:22:59,055
PERHAPS IS COMPENSATORY IN THE

1889
01:22:59,055 --> 01:23:01,358
ABSENCE OF THE MACHINERY, BUT,

1890
01:23:01,358 --> 01:23:02,492
YEAH, IT'S A HYPOTHESIS.

1891
01:23:02,492 --> 01:23:04,661
>> I THINK THERE'S A VERY

1892
01:23:04,661 --> 01:23:05,929
INTERESTING PAPER IN NEW ENGLAND

1893
01:23:05,929 --> 01:23:10,033
JOURNAL MANY YEARS AGO BY A TEAM

1894
01:23:10,033 --> 01:23:14,204
AT NIH, A COHORT OF HUNDREDS OF

1895
01:23:14,204 --> 01:23:16,239
CGD PATIENTS EVEN A SMALL

1896
01:23:16,239 --> 01:23:17,741
PERCENTAGE OF DIFFERENCE IN

1897
01:23:17,741 --> 01:23:19,876
RESIDUAL ACTIVITY OF THE ENZYME,

1898
01:23:19,876 --> 01:23:21,077
BIG EFFECT ON PHENOTYPE.

1899
01:23:21,077 --> 01:23:24,247
COULD BE INTERESTING TO

1900
01:23:24,247 --> 01:23:29,986
RECLASSIFY THE CGD PATIENTS WITH

1901
01:23:29,986 --> 01:23:33,123
AUTOIMMUNITY VIS-A-VIS RESIDUAL

1902
01:23:33,123 --> 01:23:33,390
ACTIVITY.

1903
01:23:33,390 --> 01:23:34,457
>> ONE COULD SAY THERE
COULD BE

1904
01:23:34,457 --> 01:23:37,694
MORE THAN ONE PATHWAY TO GET

1905
01:23:37,694 --> 01:23:38,595
AUTOIMMUNE DISEASE.

1906
01:23:38,595 --> 01:23:38,962
>> ABSOLUTELY.

1907
01:23:38,962 --> 01:23:44,301
>> NOT AS IF THE FACT YOU
CAN

1908
01:23:44,301 --> 01:23:49,306
GET AUTOIMMUNE DISEASE WITH CGD

1909
01:23:49,306 --> 01:23:50,440
THAT SOMEHOW DISPROVES THAT

1910
01:23:50,440 --> 01:23:52,242
NET-OSIS ISN'T IMPORTANT IN SOME

1911
01:23:52,242 --> 01:23:58,481
PATIENTS WITH AUTOIMMUNE DISEASE

1912
01:23:58,481 --> 01:23:58,915
ANYWAY.

1913
01:23:58,915 --> 01:23:59,115
OKAY.

1914
01:23:59,115 --> 01:24:05,255
SO, NOW SOME QUESTIONS WITH

1915
01:24:05,255 --> 01:24:08,058
REGARD TO THE MECHANISMS OF

1916
01:24:08,058 --> 01:24:08,491
NET-OSIS.

1917
01:24:08,491 --> 01:24:11,361
SO ONE OF THE MEMBERS OF THE

1918
01:24:11,361 --> 01:24:15,865
AUDIENCE WAS INTRIGUED BY THE

1919
01:24:15,865 --> 01:24:17,467
CONNECTION WITH GASDERMIN-D, AND

1920
01:24:17,467 --> 01:24:21,538
OF COURSE  THAT UNDER SOME

1921
01:24:21,538 --> 01:24:25,208
CIRCUMSTANCES CAN BE ACTIVATED

1922
01:24:25,208 --> 01:24:28,178
BY THE INFLAMMASOME, CASPASE 1

1923
01:24:28,178 --> 01:24:34,384
ACTIVATION CAN LEAD TO

1924
01:24:34,384 --> 01:24:35,051
GASDERMIN-D CLEAVAGE AND

1925
01:24:35,051 --> 01:24:36,119
ACTIVATION OF GASDERMIN-D.

1926
01:24:36,119 --> 01:24:38,655
THE VIEWER WANTS TO KNOW WHETHER

1927
01:24:38,655 --> 01:24:40,490
OR NOT INFLAMMASOME ACTIVATION

1928
01:24:40,490 --> 01:24:47,063
MIGHT BE RELATED TO NET-OSIS IN

1929
01:24:47,063 --> 01:24:49,933
SOME WAY.

1930
01:24:49,933 --> 01:24:54,771
>> YES, THERE IS WORK BY
KATE

1931
01:24:54,771 --> 01:24:58,341
STRUTTER IN AUSTRALIA,

1932
01:24:58,341 --> 01:24:59,376
INFLAMMASOMIVATION LEADS TO

1933
01:24:59,376 --> 01:25:01,478
NET-OSIS I THINK IN MICE AND

1934
01:25:01,478 --> 01:25:04,514
SOME HUMANS IN HER WORK.

1935
01:25:04,514 --> 01:25:08,318
TO BE COMPLETELY TRANSPARENT THE

1936
01:25:08,318 --> 01:25:09,419
LABORATORY HAD CONTRADICTORY

1937
01:25:09,419 --> 01:25:13,289
RESULTS TO OURS, AND THAT ISSUE

1938
01:25:13,289 --> 01:25:16,726
WAS NEVER COMPLETE RESULT.

1939
01:25:16,726 --> 01:25:18,995
WE HAD THAT INHIBITOR, ALSO USED

1940
01:25:18,995 --> 01:25:21,064
GASDERMIN-D KNOCKOUT MICE AND

1941
01:25:21,064 --> 01:25:24,234
SHOWED THEY CAN'T -- THE LAB

1942
01:25:24,234 --> 01:25:25,635
BELIEVES IT'S INHIBITING AN

1943
01:25:25,635 --> 01:25:29,239
ENZYME IN A PATHWAY THAT'S

1944
01:25:29,239 --> 01:25:30,673
IMPORTANT FOR THE NADPH,

1945
01:25:30,673 --> 01:25:32,442
PROBABLY TOO DETAILED FOR THIS

1946
01:25:32,442 --> 01:25:32,742
DISCUSSION.

1947
01:25:32,742 --> 01:25:35,412
THE ANSWER IN SOME CIRCUMSTANCES

1948
01:25:35,412 --> 01:25:39,482
YES, THE INFLAMMASOME CAN BE

1949
01:25:39,482 --> 01:25:39,916
ACTIVATED.

1950
01:25:39,916 --> 01:25:41,785
GASDERMIN TO MAKE NET.

1951
01:25:41,785 --> 01:25:42,886
WE THOUGHT ACTIVATION OF

1952
01:25:42,886 --> 01:25:48,925
GASDERMIN THAT HAS TO BE CLEAVED

1953
01:25:48,925 --> 01:25:49,959
IS THROUGH ELASTASE.

1954
01:25:49,959 --> 01:25:52,162
>> YOU MENTIONED THAT.

1955
01:25:52,162 --> 01:25:56,933
OF COURSE, THIS GETTING BACK TO

1956
01:25:56,933 --> 01:25:58,234
POSSIBILITY INFLAMMASOME IS

1957
01:25:58,234 --> 01:26:00,303
SOMEHOW INVOLVED UNDER SOME

1958
01:26:00,303 --> 01:26:02,138
CIRCUMSTANCES REACTIVE OXYGEN

1959
01:26:02,138 --> 01:26:05,141
SPECIES CAN INDUCE INFLAMMASOME

1960
01:26:05,141 --> 01:26:06,109
ACTIVATION.

1961
01:26:06,109 --> 01:26:09,679
AND, YOU KNOW, AT LEAST IN THE

1962
01:26:09,679 --> 01:26:11,047
SENSE THAT REACTIVE OXYGEN

1963
01:26:11,047 --> 01:26:12,982
SPECIES SEEM TO BE INVOLVED IN

1964
01:26:12,982 --> 01:26:21,958
THE PROCESS, EARLY IN THE

1965
01:26:21,958 --> 01:26:23,827
PROCESS OF NETosis.

1966
01:26:23,827 --> 01:26:28,097
THERE COULD BE DEFINITELY A

1967
01:26:28,097 --> 01:26:29,199
CONNECTION.

1968
01:26:29,199 --> 01:26:33,503
AND CERTAINLY AN AREA FOR

1969
01:26:33,503 --> 01:26:34,103
FURTHER INVESTIGATION.

1970
01:26:34,103 --> 01:26:36,005
SO ONE OF THE MEMBERS OF THE

1971
01:26:36,005 --> 01:26:39,442
AUDIENCE I THINK THIS QUESTION

1972
01:26:39,442 --> 01:26:42,812
YOU'VE ALREADY ADDRESSED IS AT

1973
01:26:42,812 --> 01:26:44,447
WHAT STAGE IN DIFFERENTIATION DO

1974
01:26:44,447 --> 01:26:46,449
NEUTROPHILS ACQUIRE ABILITY TO

1975
01:26:46,449 --> 01:26:47,116
UNDERGO NET-OSIS?

1976
01:26:47,116 --> 01:26:49,586
AND I THINK THAT MARIANA MAY

1977
01:26:49,586 --> 01:26:52,922
HAVE SAID SOMETHING ABOUT THERE

1978
01:26:52,922 --> 01:26:54,457
NEEDING TO BE RELATIVELY MATURE

1979
01:26:54,457 --> 01:26:56,626
BUT I'LL POSE THE QUESTION FOR

1980
01:26:56,626 --> 01:27:01,865
THE SAKE OF OUR AUDIENCE.

1981
01:27:01,865 --> 01:27:05,468
>> ARTURO, DO YOU WANT TO

1982
01:27:05,468 --> 01:27:05,702
ANSWER?

1983
01:27:05,702 --> 01:27:07,036
>>  MARIANA, WHY DON'T YOU

1984
01:27:07,036 --> 01:27:08,438
COMMENT.

1985
01:27:08,438 --> 01:27:09,606
>> YEAH, THERE HAVE BEEN
STUDIES

1986
01:27:09,606 --> 01:27:13,243
BY SEVERAL GROUPS THAT HAVE

1987
01:27:13,243 --> 01:27:14,511
SHOWN THAT IMMATURE NEUTROPHILS

1988
01:27:14,511 --> 01:27:18,248
ARE LESS ABLE TO FORM NETs,

1989
01:27:18,248 --> 01:27:20,049
AND INDEED IN OUR CASE WHAT I

1990
01:27:20,049 --> 01:27:21,784
MENTIONED ABOUT THE DIFFERENCES

1991
01:27:21,784 --> 01:27:22,886
BETWEEN WOMEN AND MEN AND

1992
01:27:22,886 --> 01:27:24,754
ABILITY TO FORM NETs WE THINK

1993
01:27:24,754 --> 01:27:29,893
IS IN PART DRIVEN BY THE FACT

1994
01:27:29,893 --> 01:27:31,327
THAT WOMEN'S NEUTROPHILS ARE

1995
01:27:31,327 --> 01:27:34,097
MORE MATURE, THAT MAY BE ONE

1996
01:27:34,097 --> 01:27:40,770
REASON FOR EXAMPLE JAK/STAT

1997
01:27:40,770 --> 01:27:43,039
SIGNALING APPEARS TO BE MORE

1998
01:27:43,039 --> 01:27:51,814
ROBUST IN FEMALE NEUTROPHILS,

1999
01:27:51,814 --> 01:27:54,484
MAY BE MORE CONDUCIVE TO

2000
01:27:54,484 --> 01:27:55,451
ENHANCED NET FORMATION.

2001
01:27:55,451 --> 01:27:58,555
>> NOW, ANOTHER MEMBER OF
THE

2002
01:27:58,555 --> 01:28:00,189
AUDIENCE WAS INTRIGUED BY A

2003
01:28:00,189 --> 01:28:03,927
COMMENT THAT ARTURO HAD MADE IN

2004
01:28:03,927 --> 01:28:11,367
HIS DESCRIPTION OF THE EARLY

2005
01:28:11,367 --> 01:28:20,476
PROCESS  OF NETosis AND Ki67

2006
01:28:20,476 --> 01:28:21,911
IS EXPRESSED.

2007
01:28:21,911 --> 01:28:25,281
WHAT IS Ki67 FROM A PROTEIN

2008
01:28:25,281 --> 01:28:26,849
LEVEL, WHAT'S IT DOING, AND

2009
01:28:26,849 --> 01:28:29,552
WOULD THAT HAVE ANY MECHANISTIC

2010
01:28:29,552 --> 01:28:33,823
IMPLICATIONS FOR WHAT'S GOING ON

2011
01:28:33,823 --> 01:28:34,424
IN NETosis?

2012
01:28:34,424 --> 01:28:36,059
>>  GREAT QUESTION.

2013
01:28:36,059 --> 01:28:37,794
EMBARRASSED TO SAY FOR YEARS

2014
01:28:37,794 --> 01:28:40,196
Ki67 WAS THE MARKER OF

2015
01:28:40,196 --> 01:28:41,431
NETosis WITHOUT KNOWING THE

2016
01:28:41,431 --> 01:28:43,433
FUNCTION OF THE PROTEIN.

2017
01:28:43,433 --> 01:28:46,002
IT WAS DESCRIBED A COUPLE YEARS

2018
01:28:46,002 --> 01:28:47,837
AGO BUT I'M EMBARRASSED TO SAY I

2019
01:28:47,837 --> 01:28:48,805
DON'T REMEMBER.

2020
01:28:48,805 --> 01:28:53,109
HAVE THE EXCUSE IT'S VERY LATE

2021
01:28:53,109 --> 01:28:54,210
HERE.

2022
01:28:54,210 --> 01:28:54,644
[LAUGHTER]

2023
01:28:54,644 --> 01:28:56,379
BUT WE NEVER THOUGHT THERE MIGHT

2024
01:28:56,379 --> 01:28:57,914
BE A LINK TO NETosis.

2025
01:28:57,914 --> 01:29:00,750
WE THINK IT'S A MARKER BUT WE

2026
01:29:00,750 --> 01:29:04,787
HAVEN'T DONE THE EXPERIMENTS OF

2027
01:29:04,787 --> 01:29:05,421
THE DELETING Ki67 IN

2028
01:29:05,421 --> 01:29:11,628
NEUTROPHILS TO SEE IF IT AFFECTS

2029
01:29:11,628 --> 01:29:12,595
NET FORMATION.

2030
01:29:12,595 --> 01:29:12,929
GOOD QUESTION.

2031
01:29:12,929 --> 01:29:14,197
>> THERE ARE A LOT OF
THINGS,

2032
01:29:14,197 --> 01:29:18,701
THERE ARE MARKERS, WE USE THEM

2033
01:29:18,701 --> 01:29:19,936
AS SUCH IN VARIOUS EXPERIMENTS

2034
01:29:19,936 --> 01:29:24,007
OR EVEN LOOKING AT PATIENTS, BUT

2035
01:29:24,007 --> 01:29:28,911
DON'T EXACTLY KNOW THE BIOLOGIC

2036
01:29:28,911 --> 01:29:29,545
EFFECTS.

2037
01:29:29,545 --> 01:29:36,252
SO THEN DR. KAPLAN WAS TALKING

2038
01:29:36,252 --> 01:29:37,654
ABOUT VARIOUS MUSCARINIC

2039
01:29:37,654 --> 01:29:40,923
RECEPTORS AND THEIR POSSIBLE

2040
01:29:40,923 --> 01:29:42,425
ROLE IN NETosis.

2041
01:29:42,425 --> 01:29:44,894
AND THERE'S A VIEWER WHO IS

2042
01:29:44,894 --> 01:29:49,198
INTERESTED IN THE ROAM OF THE

2043
01:29:49,198 --> 01:29:51,401
VAGUS NERVE IN INFLAMMATION AND

2044
01:29:51,401 --> 01:29:53,870
OF COURSE THERE'S THE WORK OF

2045
01:29:53,870 --> 01:29:55,538
DR. TRACY UP IN NEW YORK HAVING

2046
01:29:55,538 --> 01:29:58,608
TO DO WITH VAGUS NERVE

2047
01:29:58,608 --> 01:30:01,144
STIMULATION AND ITS EFFECT ON

2048
01:30:01,144 --> 01:30:01,477
INFLAMMATION.

2049
01:30:01,477 --> 01:30:04,013
DO YOU WANT TO MAKE ANY COMMENTS

2050
01:30:04,013 --> 01:30:06,416
WITH REGARD TO THAT WORK AND HOW

2051
01:30:06,416 --> 01:30:10,753
IT MIGHT CONNECT TO NETosis OR

2052
01:30:10,753 --> 01:30:11,187
NOT?

2053
01:30:11,187 --> 01:30:14,557
>>  YEAH, ALL THIS FIELD
OF

2054
01:30:14,557 --> 01:30:15,892
NEUROIMMUNE MODULATION, RIGHT,

2055
01:30:15,892 --> 01:30:18,594
HOW RECEPTORS COULD BE PRESENT

2056
01:30:18,594 --> 01:30:19,195
ON IMMUNE CELLS WE'RE FINDING

2057
01:30:19,195 --> 01:30:21,497
NOW NOT ONLY THEY ARE PRESENT

2058
01:30:21,497 --> 01:30:23,700
BUT SEEM TO BE FUNCTIONAL AND

2059
01:30:23,700 --> 01:30:24,701
IMPORTANT, IT'S FASCINATING.

2060
01:30:24,701 --> 01:30:28,471
I MEAN, WE HAVE PRIMARILY

2061
01:30:28,471 --> 01:30:30,573
FOCUSED ON MUSCARINIC RECEPTORS

2062
01:30:30,573 --> 01:30:34,877
AND DID SOME WORK WITH THEM AT

2063
01:30:34,877 --> 01:30:38,047
OF ON ADENOSINE AND BUT WHAT

2064
01:30:38,047 --> 01:30:38,915
ROLE ADENOSINE PLAYS IN

2065
01:30:38,915 --> 01:30:40,216
PROMOTING NET FORMATION SO WE

2066
01:30:40,216 --> 01:30:45,922
ARE COMING OUT WITH MORE AND

2067
01:30:45,922 --> 01:30:46,589
MORE NEUROIMMUNOMODULATORS

2068
01:30:46,589 --> 01:30:49,625
RELEVANT TO MYELOID CELL

2069
01:30:49,625 --> 01:30:51,260
BIOLOGY, EITHER INHIBITING OR

2070
01:30:51,260 --> 01:30:55,565
ENGAGING RECEPTORS MAY HAVE

2071
01:30:55,565 --> 01:30:56,232
POTENTIALLY PRO

2072
01:30:56,232 --> 01:30:59,402
ANTI-INFLAMMATORY, BUT ALSO

2073
01:30:59,402 --> 01:31:01,237
IMPORTANT ROLES, STILL A LOT WE

2074
01:31:01,237 --> 01:31:03,473
DON'T KNOW BUT THIS IS A WAY TO

2075
01:31:03,473 --> 01:31:04,607
SHOW HOW STUDYING PATIENTS IS

2076
01:31:04,607 --> 01:31:07,043
ALSO HELPING US GO BACK AND

2077
01:31:07,043 --> 01:31:08,544
UNDERSTAND THE BIOLOGY OF HOW

2078
01:31:08,544 --> 01:31:10,346
THESE CELLS WORK.

2079
01:31:10,346 --> 01:31:13,182
>> YEAH, YEAH.

2080
01:31:13,182 --> 01:31:14,684
OKAY.

2081
01:31:14,684 --> 01:31:15,818
SO THEN ANOTHER MECHANISTIC

2082
01:31:15,818 --> 01:31:19,889
COMMENT BUT SORT OF GETTING US

2083
01:31:19,889 --> 01:31:22,558
INTO THE WORLD OF -- REAL WORLD

2084
01:31:22,558 --> 01:31:28,364
SITUATIONS IS THE WHOLE QUESTION

2085
01:31:28,364 --> 01:31:32,268
OF PAD 4 VARIANTS, AND THE

2086
01:31:32,268 --> 01:31:36,672
CONNECTION WITH PREDELICTION TO

2087
01:31:36,672 --> 01:31:36,906
NETosis.

2088
01:31:36,906 --> 01:31:39,909
AND THE QUESTIONER IS RECALLING

2089
01:31:39,909 --> 01:31:41,844
CONNECTION BETWEEN PAD 4 AND

2090
01:31:41,844 --> 01:31:45,148
CIGARETTE SMOKING, AND THE IDEA

2091
01:31:45,148 --> 01:31:49,786
THAT PERHAPS VARIANTS IN PAD 4

2092
01:31:49,786 --> 01:31:53,055
MIGHT BE ASSOCIATED WITH

2093
01:31:53,055 --> 01:31:55,024
RHEUMATOID ARTHRITIS IN SMOKERS.

2094
01:31:55,024 --> 01:31:59,428
SO THE QUESTION IS WHETHER

2095
01:31:59,428 --> 01:32:01,164
PERHAPS THE NETosis MIGHT BE

2096
01:32:01,164 --> 01:32:04,534
THE MECHANISM BY WHICH THAT IS

2097
01:32:04,534 --> 01:32:04,967
HAPPENING.

2098
01:32:04,967 --> 01:32:07,370
>> YEAH, SO THIS IS A
REALLY

2099
01:32:07,370 --> 01:32:08,070
FASCINATING STORY.

2100
01:32:08,070 --> 01:32:10,907
I THINK ON ONE HAND IT'S BEEN

2101
01:32:10,907 --> 01:32:17,380
SHOWN THAT SMOKING BY ITSELF CAN

2102
01:32:17,380 --> 01:32:19,048
LEAD TO SITRILINATION OF

2103
01:32:19,048 --> 01:32:20,883
TISSUES, THE LINK MAY BE PERHAPS

2104
01:32:20,883 --> 01:32:27,423
IN PART DRIVEN BY THE FACT IF

2105
01:32:27,423 --> 01:32:31,194
YOU SMOKE THAN IF YOU HAVE

2106
01:32:31,194 --> 01:32:32,361
GENETIC PREDISPOSITION MAY BE

2107
01:32:32,361 --> 01:32:33,763
ONE MECHANISM.

2108
01:32:33,763 --> 01:32:42,805
IT'S SHOWN SMOKING MAY TRIGGER

2109
01:32:42,805 --> 01:32:44,473
NET FORMATION, PERFECT STORM,

2110
01:32:44,473 --> 01:32:48,311
THINGS HAPPENING IN MUCOSA OF

2111
01:32:48,311 --> 01:32:53,583
THE LUNGS, GENETICALLY DRIVEN OR

2112
01:32:53,583 --> 01:32:55,384
IF SOMEONE SMOKES, SITRILINASE,

2113
01:32:55,384 --> 01:32:55,852
MAY GET AMPLIFIED.

2114
01:32:55,852 --> 01:32:59,055
THERE ARE A NUMBER OF GENETIC

2115
01:32:59,055 --> 01:33:00,489
POLYMORPHISMS ASSOCIATED TO

2116
01:33:00,489 --> 01:33:02,024
IMMUNITY THAT HAVE BEEN FOUND TO

2117
01:33:02,024 --> 01:33:04,527
LEAD TO ENHANCE THE NET

2118
01:33:04,527 --> 01:33:07,430
FORMATION EVEN FORM AUTOIMMUNITY

2119
01:33:07,430 --> 01:33:09,332
DEVELOPS, AND ONE IS RELATED TO

2120
01:33:09,332 --> 01:33:10,433
PRODUCTIVITY BUT THERE ARE

2121
01:33:10,433 --> 01:33:13,502
OTHERS WHICH MAKE IT FASCINATING

2122
01:33:13,502 --> 01:33:16,706
HOW ENVIRONMENT AND GENETICS MAY

2123
01:33:16,706 --> 01:33:18,341
WORK TOGETHER TO INDUCE

2124
01:33:18,341 --> 01:33:23,212
DYSREGULATION EVEN BEFORE

2125
01:33:23,212 --> 01:33:23,779
AUTOIMMUNITY DEVELOPS.

2126
01:33:23,779 --> 01:33:27,550
>> FROM THE  OTHER SIDE OF
THE

2127
01:33:27,550 --> 01:33:30,386
GENETIC WORLD, ONE OF OUR

2128
01:33:30,386 --> 01:33:31,153
AUDIENCE WAS INTERESTED IN

2129
01:33:31,153 --> 01:33:35,758
LEARNING A LITTLE BIT MORE ABOUT

2130
01:33:35,758 --> 01:33:42,365
HOW MICROING  MICROORGANISMS MAE

2131
01:33:42,365 --> 01:33:45,201
EVOLVED STRATEGIES TO SABOTAGE

2132
01:33:45,201 --> 01:33:48,804
NETosis, TO PREVENT OR

2133
01:33:48,804 --> 01:33:49,138
COUNTERACT IT.

2134
01:33:49,138 --> 01:33:50,673
I SUPPOSE NUCLEASES BEING

2135
01:33:50,673 --> 01:33:53,075
PRODUCED BY BACTERIA WOULD BE

2136
01:33:53,075 --> 01:33:56,145
ONE SUCH MECHANISM, BUT CAN YOU

2137
01:33:56,145 --> 01:33:59,415
MAYBE TALK ABOUT ANY OTHERS THAT

2138
01:33:59,415 --> 01:34:04,820
MAY HAVE BEEN LOOKED AT?

2139
01:34:04,820 --> 01:34:07,990
>>  WELL, CATALASE
CONSUMES

2140
01:34:07,990 --> 01:34:10,493
HYDROGEN PEROXIDE, PREVENTS NET

2141
01:34:10,493 --> 01:34:13,095
FORMATION, THAT'S ANOTHER.

2142
01:34:13,095 --> 01:34:19,402
NUCLEASES ARE ALSO FOR EXAMPLE

2143
01:34:19,402 --> 01:34:20,303
PRODUCED BY PARASITES.

2144
01:34:20,303 --> 01:34:25,308
THOSE ARE THE TWO THAT COME TO

2145
01:34:25,308 --> 01:34:25,608
MIND.

2146
01:34:25,608 --> 01:34:28,044
I'LL THINK -- I THINK WE WROTE A

2147
01:34:28,044 --> 01:34:30,146
REVIEW ABOUT THIS AT SOME POINT

2148
01:34:30,146 --> 01:34:33,683
A FEW YEARS AGO SO I'LL HAVE TO

2149
01:34:33,683 --> 01:34:35,151
RE-READ IT.

2150
01:34:35,151 --> 01:34:35,451
>> BUT --

2151
01:34:35,451 --> 01:34:39,555
>>  MARIANA, DO YOU
REMEMBER?

2152
01:34:39,555 --> 01:34:48,597
>>  NETosis IS A BIG ONE.

2153
01:34:48,597 --> 01:34:53,669
>> THAT MAKES SENSE, IN
FACT I

2154
01:34:53,669 --> 01:34:57,540
GUESS THAT'S EVEN PART OF THE

2155
01:34:57,540 --> 01:35:00,343
REASON WHY IN BACTERIOLOGY WE

2156
01:35:00,343 --> 01:35:02,111
HAVE CATALASE POSITIVE AND

2157
01:35:02,111 --> 01:35:03,846
CATALASE NEGATIVE AND SO FORTH

2158
01:35:03,846 --> 01:35:05,047
BACTERIA, SORT OF DENOTING THEM

2159
01:35:05,047 --> 01:35:07,984
BASED ON WHETHER THEY HAVE

2160
01:35:07,984 --> 01:35:15,024
EVOLVED THOSE MECHANISMS OR NOT.

2161
01:35:15,024 --> 01:35:15,224
OKAY.

2162
01:35:15,224 --> 01:35:17,593
SO, LET'S SEE.

2163
01:35:17,593 --> 01:35:19,428
HERE'S SOMEONE, A HOPEFUL MEMBER

2164
01:35:19,428 --> 01:35:23,399
OF THE AUDIENCE, FOR THE

2165
01:35:23,399 --> 01:35:25,234
NEUTROPHILS I GUESS, IS THERE

2166
01:35:25,234 --> 01:35:29,138
ANY WAY THAT NEUTROPHILS COULD

2167
01:35:29,138 --> 01:35:32,274
FORM NETs WITHOUT DYING?

2168
01:35:32,274 --> 01:35:37,613
>> SO THIS HAS BEEN AN
AREA OF A

2169
01:35:37,613 --> 01:35:39,448
LOT OF CONTROVERSY, I THINK.

2170
01:35:39,448 --> 01:35:42,952
THERE HAVE BEEN GROUPS THAT HAVE

2171
01:35:42,952 --> 01:35:44,620
SUGGESTED UNDER CERTAIN

2172
01:35:44,620 --> 01:35:48,224
CIRCUMSTANCES UPON EXPOSURE TO

2173
01:35:48,224 --> 01:35:49,692
CERTAIN MICROBES NEUTROPHIL MAY

2174
01:35:49,692 --> 01:35:53,696
BE ABLE TO FORM NETs WITHOUT

2175
01:35:53,696 --> 01:35:54,930
DYING.

2176
01:35:54,930 --> 01:35:57,433
AND KIND OF EXTRUDE THE NUCLEUS

2177
01:35:57,433 --> 01:36:00,870
BUT CONTINUE TO FUNCTION AND

2178
01:36:00,870 --> 01:36:04,173
PERFORM VARIOUS ACTIVITIES.

2179
01:36:04,173 --> 01:36:06,809
IN OUR HANDS, NETs FORM DURING

2180
01:36:06,809 --> 01:36:07,910
AUTOIMMUNE DISEASES, WHAT WE

2181
01:36:07,910 --> 01:36:08,744
CALL SUICIDAL.

2182
01:36:08,744 --> 01:36:14,316
WE ALWAYS SEE THAT THE CELL

2183
01:36:14,316 --> 01:36:15,484
DIES, NO DISRUPTION OF CELL

2184
01:36:15,484 --> 01:36:20,890
MEMBRANE AND IT'S A FINAL EVENT 

2185
01:36:20,890 --> 01:36:21,891
BUT ARTURO I'M SURE CAN TALK

2186
01:36:21,891 --> 01:36:23,025
ABOUT THIS.

2187
01:36:23,025 --> 01:36:27,630
>> I'LL SAY THE SAME
THING.

2188
01:36:27,630 --> 01:36:29,365
THERE ARE REPORTS IT WOULD BE

2189
01:36:29,365 --> 01:36:31,100
VERY INTERESTING TO FIND IF

2190
01:36:31,100 --> 01:36:32,468
THERE'S A MECHANISM SO WE CAN

2191
01:36:32,468 --> 01:36:34,937
INTERRUPT IT AND SEE HOW IT

2192
01:36:34,937 --> 01:36:35,137
WORKS.

2193
01:36:35,137 --> 01:36:38,841
SO, THAT'S WHY IT'S IMPORTANT IN

2194
01:36:38,841 --> 01:36:41,944
WORK ON MECHANISMS TO SEE, TO

2195
01:36:41,944 --> 01:36:44,246
IDENTIFY FUNCTION AND TO

2196
01:36:44,246 --> 01:36:50,252
INTERRUPT AND I WILL SEE WHEN

2197
01:36:50,252 --> 01:36:51,454
THOSE APPEAR AND DECIDE.

2198
01:36:51,454 --> 01:36:52,721
>> IF IT DID HAPPEN, THERE
WOULD

2199
01:36:52,721 --> 01:36:56,492
HAVE TO BE SOME SORT OF GHOST

2200
01:36:56,492 --> 01:36:58,027
CELLS OR WHATEVER THAT WOULD BE

2201
01:36:58,027 --> 01:37:04,834
THE SORT OF THE REMNANTS OF THE

2202
01:37:04,834 --> 01:37:06,502
NEUTROPHIL THAT'S EXTRUDED ITS

2203
01:37:06,502 --> 01:37:09,105
NUCLEIC ACIDS BUT STILL HAS SOME

2204
01:37:09,105 --> 01:37:11,173
FUNCTION OR AT LEAST THERE'S THE

2205
01:37:11,173 --> 01:37:14,710
CARCASS OF IT GOING AROUND IN

2206
01:37:14,710 --> 01:37:15,077
THE CIRCULATION.

2207
01:37:15,077 --> 01:37:19,248
SO, WELL, AN INTERESTING THING

2208
01:37:19,248 --> 01:37:20,349
TO LOOK FOR.

2209
01:37:20,349 --> 01:37:23,219
AND WE'LL AWAIT THE NEXT

2210
01:37:23,219 --> 01:37:23,819
PRESENTATION IN DEMYSTIFYING

2211
01:37:23,819 --> 01:37:29,258
MEDICINE TO HEAR ABOUT THAT I

2212
01:37:29,258 --> 01:37:29,458
GUESS.

2213
01:37:29,458 --> 01:37:32,661
SO, ANOTHER QUESTION, HOW IS

2214
01:37:32,661 --> 01:37:34,163
NETosis REGULATED?

2215
01:37:34,163 --> 01:37:36,532
ARE THERE SPECIFIC CYTOKINES

2216
01:37:36,532 --> 01:37:38,734
THAT POTENTIATE OR INHIBIT

2217
01:37:38,734 --> 01:37:38,968
NETosis?

2218
01:37:38,968 --> 01:37:43,072
I THINK THAT IN ONE OF THE

2219
01:37:43,072 --> 01:37:45,674
MARIANA'S DIAGRAMS THERE WERE

2220
01:37:45,674 --> 01:37:46,609
SOME CYTOKINES THAT WERE

2221
01:37:46,609 --> 01:37:48,043
INDICATED THERE BUT MAYBE YOU

2222
01:37:48,043 --> 01:37:50,613
COULD REVIEW THEM FOR THE

2223
01:37:50,613 --> 01:37:50,946
VIEWER.

2224
01:37:50,946 --> 01:37:53,849
>> YES, I MEAN, SOME THAT
COME

2225
01:37:53,849 --> 01:37:57,486
TO MIND TNF WE'VE FOUND IS A

2226
01:37:57,486 --> 01:38:02,925
STRONG INDUCER OF NETs AND WE

2227
01:38:02,925 --> 01:38:04,460
WEEK IN RHEUMATOID SYNOVIUM.

2228
01:38:04,460 --> 01:38:06,729
IL-17 APPEARS TO HAVE SOME

2229
01:38:06,729 --> 01:38:08,397
EFFECT ON NET FORMATION, IT'S

2230
01:38:08,397 --> 01:38:14,036
BEEN DESCRIBED IN OUR HANDS IT'S

2231
01:38:14,036 --> 01:38:17,173
PRETTY WEAK BUT INDUCER OF

2232
01:38:17,173 --> 01:38:17,373
NETs.

2233
01:38:17,373 --> 01:38:17,973
CYTOKINES THAT INHIBIT I DON'T

2234
01:38:17,973 --> 01:38:19,575
THINK THERE'S ONE THAT COMES TO

2235
01:38:19,575 --> 01:38:22,845
MIND THAT'S BEEN FOUND TO HAVE

2236
01:38:22,845 --> 01:38:23,646
INHIBITORY EFFECTS ON ABILITY TO

2237
01:38:23,646 --> 01:38:25,047
FORM NETs.

2238
01:38:25,047 --> 01:38:27,950
MANY ARE TARGETS GOING TO THE

2239
01:38:27,950 --> 01:38:35,024
JAK/STAT PATHWAY BUT NOT ONLY,

2240
01:38:35,024 --> 01:38:35,224
RIGHT?

2241
01:38:35,224 --> 01:38:37,793
BUT CNS IS A BIG ONE.

2242
01:38:37,793 --> 01:38:38,961
>> THEN THAT WOULD ALSO
PROVIDE

2243
01:38:38,961 --> 01:38:43,499
AT LEAST ONE OF THE MECHANISMS

2244
01:38:43,499 --> 01:38:45,768
BY WHICH TNF INHIBITORS FOR

2245
01:38:45,768 --> 01:38:50,072
EXAMPLE WORK IN CERTAIN

2246
01:38:50,072 --> 01:38:52,241
AUTOIMMUNE DISEASES ANYWAY.

2247
01:38:52,241 --> 01:38:55,211
SO THAT WOULD BE SATISFYING IN

2248
01:38:55,211 --> 01:38:59,915
TERMS OF KNOWING ABOUT THAT.

2249
01:38:59,915 --> 01:39:00,583
OKAY.

2250
01:39:00,583 --> 01:39:03,352
SO, ANOTHER MEMBER OF THE

2251
01:39:03,352 --> 01:39:04,954
AUDIENCE WANTS TO KNOW ABOUT

2252
01:39:04,954 --> 01:39:06,822
NEUTROPHIL NETs AND WHETHER

2253
01:39:06,822 --> 01:39:14,396
THEY PLAY A ROLE IN GOUT.

2254
01:39:14,396 --> 01:39:14,663
>> YES.

2255
01:39:14,663 --> 01:39:17,900
YOU GO AHEAD.

2256
01:39:17,900 --> 01:39:18,467
IT'S AUTOIMMUNE,

2257
01:39:18,467 --> 01:39:19,935
AUTO-INFLAMMATORY DISEASE.

2258
01:39:19,935 --> 01:39:25,040
YES, THERE'S VERY GOOD DATA FROM

2259
01:39:25,040 --> 01:39:26,475
GERMANY.

2260
01:39:26,475 --> 01:39:29,011
>> YES, IT WAS FOUND THAT
IT'S

2261
01:39:29,011 --> 01:39:29,912
ONE OF THE MOST NEUTROPHILIC

2262
01:39:29,912 --> 01:39:32,348
CONDITIONS WE CAN FIND IN THE

2263
01:39:32,348 --> 01:39:36,485
SYNOVIUM AND THERE ARE'S SO MANY

2264
01:39:36,485 --> 01:39:38,087
NEUTROPHILS GOING IN WHEN GOUT

2265
01:39:38,087 --> 01:39:43,892
OCCURS IN RESPONSE TO URIC ACID,

2266
01:39:43,892 --> 01:39:45,127
INDUCERS OF NETs, BUT THERE'S

2267
01:39:45,127 --> 01:39:49,365
SO MANY NEUTROPHILS THERE THAT

2268
01:39:49,365 --> 01:39:50,733
THE NETS BECOME AGGREGATED AND

2269
01:39:50,733 --> 01:39:52,868
SO THESE NETs ACTUALLY NOT

2270
01:39:52,868 --> 01:39:54,536
ONLY HAVE INFLAMMATORY EFFECT

2271
01:39:54,536 --> 01:39:55,604
BUT EVENTUALLY LEAD TO

2272
01:39:55,604 --> 01:39:56,605
RESOLUTION OF THE INFLAMMATION

2273
01:39:56,605 --> 01:39:59,241
WHICH WE KNOW IS A PICTURE OF

2274
01:39:59,241 --> 01:39:59,541
GOUT.

2275
01:39:59,541 --> 01:40:01,343
GOUT WILL GO AWAY EVEN IF YOU

2276
01:40:01,343 --> 01:40:04,713
DON'T TREAT IT.

2277
01:40:04,713 --> 01:40:06,215
IT'S BELIEVES AGGREGATED NETs

2278
01:40:06,215 --> 01:40:09,285
ARE CHOKING CYTOKINES AND

2279
01:40:09,285 --> 01:40:11,253
CHEMOKINES AND MAKING THE --

2280
01:40:11,253 --> 01:40:12,388
DAMPENING OR LEADING TO

2281
01:40:12,388 --> 01:40:13,822
RESOLUTION OF INFLAMMATION SO

2282
01:40:13,822 --> 01:40:17,760
IT'S A STORY OF HOW THE NET CAN

2283
01:40:17,760 --> 01:40:18,761
HAVE A VERY PRO-INFLAMMATORY

2284
01:40:18,761 --> 01:40:20,596
EFFECT BUT IN BIG QUANTITY CAN

2285
01:40:20,596 --> 01:40:24,700
LEAD TO RESOLUTION OF

2286
01:40:24,700 --> 01:40:25,034
INFLAMMATION.

2287
01:40:25,034 --> 01:40:26,435
>> WELL, OF COURSE, JUST

2288
01:40:26,435 --> 01:40:27,903
BRINGING UP THE QUESTION OF

2289
01:40:27,903 --> 01:40:32,241
GOUT, YOU KNOW, THERE ARE ALL OF

2290
01:40:32,241 --> 01:40:35,411
THE DATA WITH REGARD TO DIETARY

2291
01:40:35,411 --> 01:40:38,213
FACTORS AND HOW THEY CAN

2292
01:40:38,213 --> 01:40:40,949
SOMETIMES HAVE AN EFFECT ON

2293
01:40:40,949 --> 01:40:45,487
ACUTE GOUT ATTACKS.

2294
01:40:45,487 --> 01:40:47,856
MAYBE JUST TO RIFF ON THAT, ARE

2295
01:40:47,856 --> 01:40:51,427
THERE DIETARY FACTORS THAT IN

2296
01:40:51,427 --> 01:40:56,732
SOME WAY INCREASE OR DECREASE

2297
01:40:56,732 --> 01:40:57,399
ONE'S PREDISPOSITION FOR

2298
01:40:57,399 --> 01:40:57,633
NETosis?

2299
01:40:57,633 --> 01:41:01,870
>>  NOT THAT I KNOW OF.

2300
01:41:01,870 --> 01:41:02,104
MARIANA?

2301
01:41:02,104 --> 01:41:04,273
>>  GREAT QUESTION.

2302
01:41:04,273 --> 01:41:07,409
NOTHING THAT COMES TO MIND, YOU

2303
01:41:07,409 --> 01:41:08,577
KNOW, THAT'S A WELL-KNOWN

2304
01:41:08,577 --> 01:41:09,178
TRIGGER.

2305
01:41:09,178 --> 01:41:16,552
I WANT TO MENTION WITH GOUT IT

2306
01:41:16,552 --> 01:41:19,421
HAS BEEN SHOWING COALCHISENE,

2307
01:41:19,421 --> 01:41:25,527
BECAUSE OF MITOSIS, IT CAN

2308
01:41:25,527 --> 01:41:30,399
INHIBIT NETs, COULD BE

2309
01:41:30,399 --> 01:41:33,602
ADDITIONAL MECHANISM BY WHICH IT

2310
01:41:33,602 --> 01:41:36,605
IS EFFECTIVE IN GOUT.

2311
01:41:36,605 --> 01:41:37,473
>> ANOTHER PHARMACOLOGIC

2312
01:41:37,473 --> 01:41:38,040
INTERVENTION THAT ONE COULD

2313
01:41:38,040 --> 01:41:42,778
THINK ABOUT THAT I GUESS ARTURO

2314
01:41:42,778 --> 01:41:45,347
AT LEAST TOUCHED UPON IS THE

2315
01:41:45,347 --> 01:41:47,216
QUESTION OF STING, AND I BELIEVE

2316
01:41:47,216 --> 01:41:48,217
THAT THERE'S QUITE A GREAT

2317
01:41:48,217 --> 01:41:52,054
INTEREST NOW IN THE DEVELOPMENT

2318
01:41:52,054 --> 01:41:55,557
OF STING INHIBITORS IN THE

2319
01:41:55,557 --> 01:41:56,558
BIOTECH SECTOR.

2320
01:41:56,558 --> 01:41:59,862
ANY WORDS ON THAT WITH REGARD TO

2321
01:41:59,862 --> 01:42:05,534
POSSIBLE APPLICATION IN THESE

2322
01:42:05,534 --> 01:42:06,402
CONDITIONS?

2323
01:42:06,402 --> 01:42:07,503
>>  I DON'T HAVE ANY
INSIGHT

2324
01:42:07,503 --> 01:42:08,003
INTO THAT.

2325
01:42:08,003 --> 01:42:10,672
I THINK IT'S A VERY GOOD IDEA

2326
01:42:10,672 --> 01:42:12,975
BUT I HAVEN'T HEARD ANYTHING.

2327
01:42:12,975 --> 01:42:13,475
>> YEAH.

2328
01:42:13,475 --> 01:42:19,915
>> I MEAN, WHAT WE HAD
SEEN

2329
01:42:19,915 --> 01:42:22,818
PATIENTS THAT RECEIVE

2330
01:42:22,818 --> 01:42:24,820
AMIPROMYRAB WHICH BLOCKS TYPE 1

2331
01:42:24,820 --> 01:42:26,588
INTERFERONS, NOT DIRECTLY LINKED

2332
01:42:26,588 --> 01:42:29,224
TO STING BUT TO THE TYPE 1

2333
01:42:29,224 --> 01:42:30,526
PATHWAY DOWNSTREAM OF STING

2334
01:42:30,526 --> 01:42:33,429
APPEARS TO BE DECREASE IN NETs

2335
01:42:33,429 --> 01:42:43,839
IN THOSE PATIENTS WITH

2336
01:42:43,839 --> 01:42:44,406
INHIBITION.

2337
01:42:44,406 --> 01:42:46,809
>> I SEE DR. ARIAS TURNED
ON HIS

2338
01:42:46,809 --> 01:42:48,177
CAMERA WHICH MAY INDICATE THAT

2339
01:42:48,177 --> 01:42:49,845
HE HAS A QUESTION THAT HE WOULD

2340
01:42:49,845 --> 01:42:51,246
LIKE TO POSE.

2341
01:42:51,246 --> 01:42:54,950
WIN, DO YOU HAVE A QUESTION FOR

2342
01:42:54,950 --> 01:42:55,451
OUR SPEAKERS?

2343
01:42:55,451 --> 01:42:55,984
>>  SURE.

2344
01:42:55,984 --> 01:42:58,520
FIRST OF ALL, THANK YOU VERY

2345
01:42:58,520 --> 01:43:05,227
MUCH FOR THIS VERY SUCCESSFUL

2346
01:43:05,227 --> 01:43:06,628
PRESENTATION OF BRIDGING SCIENCE

2347
01:43:06,628 --> 01:43:07,196
AND MEDICINE.

2348
01:43:07,196 --> 01:43:14,403
SO, I HAVE TWO QUESTIONS.

2349
01:43:14,403 --> 01:43:16,472
ONE IS STEROIDS,

2350
01:43:16,472 --> 01:43:19,975
CORTICOSTEROIDS, WIDELY USED IN

2351
01:43:19,975 --> 01:43:22,077
AUTOIMMUNE DISEASES WEREN'T

2352
01:43:22,077 --> 01:43:25,147
MENTIONED IN YOUR PRESENTATIONS.

2353
01:43:25,147 --> 01:43:28,317
DO CORTICOSTEROIDS PLAY ANY ROLE

2354
01:43:28,317 --> 01:43:30,285
IN NETs FORMATION?

2355
01:43:30,285 --> 01:43:33,455
AND THE SECOND QUESTION I HAVE

2356
01:43:33,455 --> 01:43:35,557
IS I'M A LITTLE CONFUSED BY SOME

2357
01:43:35,557 --> 01:43:38,727
OF THE DIAGRAMS.

2358
01:43:38,727 --> 01:43:40,028
ARE NETs STRICTLY

2359
01:43:40,028 --> 01:43:42,331
EXTRACELLULAR OR IN SOME

2360
01:43:42,331 --> 01:43:44,700
DIAGRAMS LOOKS LIKE THEY ARE AT

2361
01:43:44,700 --> 01:43:49,271
LEAST INITIALLY COVERED BY A

2362
01:43:49,271 --> 01:43:49,805
PLASMA MEMBRANE.

2363
01:43:49,805 --> 01:43:53,175
IS MUCH KNOWN ABOUT SORT OF THE

2364
01:43:53,175 --> 01:43:56,812
BASIC CELL BIOLOGY REGARDING THE

2365
01:43:56,812 --> 01:44:01,483
FORMATION OF NETs AND IF IT

2366
01:44:01,483 --> 01:44:03,285
INVOLVES EXTENSION OF PLASMA

2367
01:44:03,285 --> 01:44:05,354
MEMBRANE, MY GOSH, THERE MUST BE

2368
01:44:05,354 --> 01:44:06,355
AN ENORMOUS AMOUNT OF CHANGES

2369
01:44:06,355 --> 01:44:09,091
TAKING PLACE WITHIN THE CELL

2370
01:44:09,091 --> 01:44:11,426
OTHER THAN JUST PHENOMENA THAT

2371
01:44:11,426 --> 01:44:12,694
YOU DESCRIBED.

2372
01:44:12,694 --> 01:44:16,999
I'M CURIOUS WHETHER CELL

2373
01:44:16,999 --> 01:44:27,476
BIOLOGISTS HAVE JUMPED INTO

2374
01:44:31,480 --> 01:44:32,214
THIS.

2375
01:44:32,214 --> 01:44:35,083
YOU MENTIONED COALCHISENE.

2376
01:44:35,083 --> 01:44:38,020
AT ANY RATE, THIS HAS BEEN AN

2377
01:44:38,020 --> 01:44:40,789
EXTREMELY EXCITING AND WE

2378
01:44:40,789 --> 01:44:41,924
APPRECIATE YOUR PARTICIPATION

2379
01:44:41,924 --> 01:44:45,827
IMMENSELY.

2380
01:44:45,827 --> 01:44:46,161
 

2381
01:44:46,161 --> 01:44:48,964
>> MAYBE I CAN ANSWER, AND
YOU

2382
01:44:48,964 --> 01:44:50,899
ADDRESS THE SECOND ONE.

2383
01:44:50,899 --> 01:44:53,068
>> YOU GO.

2384
01:44:53,068 --> 01:44:54,269
>> IT'S REALLY
FASCINATING.

2385
01:44:54,269 --> 01:44:59,641
AND THERE WAS A RECENT PAPER BY

2386
01:44:59,641 --> 01:45:02,778
SOMEONE WHO WORKS ON NETs IN

2387
01:45:02,778 --> 01:45:03,445
CANCER.

2388
01:45:03,445 --> 01:45:08,517
THERE IS THIS OBSERVATION THAT

2389
01:45:08,517 --> 01:45:10,786
OF A DRIVER OF METASTASIS IN A

2390
01:45:10,786 --> 01:45:14,823
POORLY UNDERSTOOD AWAY BUT SHE

2391
01:45:14,823 --> 01:45:15,524
MADE A CONNECTION

2392
01:45:15,524 --> 01:45:16,124
CORTICOSTEROIDS PRODUCED FOR

2393
01:45:16,124 --> 01:45:18,860
EXAMPLE UNDER CONDITIONS OF

2394
01:45:18,860 --> 01:45:19,428
STRESS ARE ENHANCING NET

2395
01:45:19,428 --> 01:45:23,231
FORMATION AND THESE MAY BE A

2396
01:45:23,231 --> 01:45:29,871
MECHANISM BECAUSE KNITS  -- NETE

2397
01:45:29,871 --> 01:45:36,144
BEEN IMPLICATED IN  METASTASIS,

2398
01:45:36,144 --> 01:45:40,015
STRESS MAY BE AN ENHANCER.

2399
01:45:40,015 --> 01:45:43,418
AT LEAST IN THE CONTEXT OF

2400
01:45:43,418 --> 01:45:44,486
CANCER, HER OBSERVATIONS

2401
01:45:44,486 --> 01:45:47,222
STEROIDS IF ANYTHING INCREASE

2402
01:45:47,222 --> 01:45:47,422
NETs.

2403
01:45:47,422 --> 01:45:49,625
NOW, THIS COULD BE -- THERE'S

2404
01:45:49,625 --> 01:45:53,595
MORE TO IT, DEPENDS ON THE

2405
01:45:53,595 --> 01:45:54,796
DOSES, CHRONICITY OF EXPOSURE,

2406
01:45:54,796 --> 01:45:57,199
BUT AT LEAST IN THE CONTEXT OF

2407
01:45:57,199 --> 01:46:00,168
CANCER IF ANYTHING IT'S THE

2408
01:46:00,168 --> 01:46:04,072
OTHER WAY OF IMMUNOSUPPRESSIVE

2409
01:46:04,072 --> 01:46:05,007
IN AUTOIMMUNITY.

2410
01:46:05,007 --> 01:46:06,675
>> JUST THINKING ABOUT
THAT FOR

2411
01:46:06,675 --> 01:46:08,610
A MOMENT, IF I CAN JUST COMMENT

2412
01:46:08,610 --> 01:46:10,779
ON YOUR COMMENT FOR A SECOND,

2413
01:46:10,779 --> 01:46:15,717
AND THAT IS THAT MAYBE

2414
01:46:15,717 --> 01:46:18,487
GLUCOCORTICOIDS COULD BE

2415
01:46:18,487 --> 01:46:20,622
MOBILIZING SUBSET OF NEUTROPHILS

2416
01:46:20,622 --> 01:46:24,660
PRONE TO MAKE NETs, YOU KNOW,

2417
01:46:24,660 --> 01:46:26,228
DEMARGINNATING A SUBPOPULATION

2418
01:46:26,228 --> 01:46:29,264
OR WHATEVER THAT'S NET PRONE.

2419
01:46:29,264 --> 01:46:31,433
>> YEAH, ABSOLUTELY.

2420
01:46:31,433 --> 01:46:35,604
I THINK, YEAH, MORE NUANCE, YES,

2421
01:46:35,604 --> 01:46:36,071
ABSOLUTELY, I AGREE.

2422
01:46:36,071 --> 01:46:38,807
>> AND TO THE FIRST PART
OF THE

2423
01:46:38,807 --> 01:46:40,676
QUESTION, I AGREE WE SHOULD --

2424
01:46:40,676 --> 01:46:42,244
WE HOPE THAT MORE CELL

2425
01:46:42,244 --> 01:46:43,211
BIOLOGISTS GET INTERESTED IN

2426
01:46:43,211 --> 01:46:43,745
NETs.

2427
01:46:43,745 --> 01:46:46,114
THE PROBLEM OF WORKING IN NETs

2428
01:46:46,114 --> 01:46:48,684
IS THAT THERE IS NOT CELL LINES

2429
01:46:48,684 --> 01:46:50,352
THAT MIMICS ACTIVITY OF NETs.

2430
01:46:50,352 --> 01:46:54,222
THERE'S NO CELL LINE THAT MAKES

2431
01:46:54,222 --> 01:46:54,556
NETs.

2432
01:46:54,556 --> 01:46:58,694
ALL THE WORK THAT HAS TO BE DONE

2433
01:46:58,694 --> 01:46:59,361
WITH REAL NEUTROPHILS AS MARIANA

2434
01:46:59,361 --> 01:47:01,496
MENTIONED THE MOUSE IS A LOUSY

2435
01:47:01,496 --> 01:47:03,565
MODEL FOR NEUTROPHIL BIOLOGY.

2436
01:47:03,565 --> 01:47:05,967
THE RATS ARE BETTER.

2437
01:47:05,967 --> 01:47:07,402
RABBITS ARE BETTER.

2438
01:47:07,402 --> 01:47:08,537
THEIR NEUTROPHILS EXPRESS MORE

2439
01:47:08,537 --> 01:47:11,606
OF THE PROTEINS WE KNOW HUMAN

2440
01:47:11,606 --> 01:47:13,175
NEUTROPHILS WORK, SO A CELL THAT

2441
01:47:13,175 --> 01:47:15,444
THERE IS NO MODEL FOR AND THAT

2442
01:47:15,444 --> 01:47:17,045
LEAVES ONLY SIX HOURS AFTER

2443
01:47:17,045 --> 01:47:19,114
TAKING IT OUT OF BLOOD IS KIND

2444
01:47:19,114 --> 01:47:21,883
OF NOT AN EASY MODEL, RIGHT?

2445
01:47:21,883 --> 01:47:24,052
ONE HAS TO REINVENT BIOLOGY

2446
01:47:24,052 --> 01:47:25,053
WITHOUT THE BENEFITS OF

2447
01:47:25,053 --> 01:47:26,655
MOLECULAR BIOLOGY.

2448
01:47:26,655 --> 01:47:28,990
IT WOULD BE GREAT TO RECRUIT

2449
01:47:28,990 --> 01:47:32,794
MORE BIOLOGISTS TO THE FIELD.

2450
01:47:32,794 --> 01:47:35,430
DO YOU AGREE, MARIANA?

2451
01:47:35,430 --> 01:47:38,800
>>  TOTALLY.

2452
01:47:38,800 --> 01:47:40,736
>> ALL RIGHT.

2453
01:47:40,736 --> 01:47:43,438
NOW WE HAVE A QUESTION FROM

2454
01:47:43,438 --> 01:47:45,073
ANOTHER MEMBER OF THE AUDIENCE

2455
01:47:45,073 --> 01:47:46,875
WITH REGARD TO A DIFFERENT FORM

2456
01:47:46,875 --> 01:47:48,276
OF CELL DEATH.

2457
01:47:48,276 --> 01:47:50,145
SO, THE QUESTIONER WANTS TO KNOW

2458
01:47:50,145 --> 01:47:56,818
DO NETs HAVE A ROLE IN

2459
01:47:56,818 --> 01:47:57,452
APOPTOSIS?

2460
01:47:57,452 --> 01:48:04,259
>> WELL, WHAT I HAVE READ
A

2461
01:48:04,259 --> 01:48:06,995
NEUTROPHIL BY APOPTOSIS OR

2462
01:48:06,995 --> 01:48:08,797
NETs, AND THE AKT PATHWAY WAS

2463
01:48:08,797 --> 01:48:09,965
SORT OF REGULATOR ON WHICH WAY

2464
01:48:09,965 --> 01:48:14,836
IT WAS GOING TO GO, BUT AS

2465
01:48:14,836 --> 01:48:19,441
ARTURO SHOWED WE SEE AT THE

2466
01:48:19,441 --> 01:48:22,677
BEGINNING, IT'S THE OPPOSITE, IN

2467
01:48:22,677 --> 01:48:24,079
APOPTOSIS SEE CHROMATIN, IN

2468
01:48:24,079 --> 01:48:25,480
NECROSIS SEE CONDENSATION SO I'M

2469
01:48:25,480 --> 01:48:33,088
NOT AWARE THAT BOTH -- I ASSUME

2470
01:48:33,088 --> 01:48:35,123
UPON SPECIFIC SIMILAR WILL DIE

2471
01:48:35,123 --> 01:48:37,425
BY NETosis, A MORE SPECIFIC

2472
01:48:37,425 --> 01:48:40,262
PROCESS THAN THE REGULAR WAY

2473
01:48:40,262 --> 01:48:41,463
BOTH NEUTROPHILS WILL DIE.

2474
01:48:41,463 --> 01:48:41,797
>> I AGREE.

2475
01:48:41,797 --> 01:48:43,799
IF THE QUESTION WAS WHETHER YOU

2476
01:48:43,799 --> 01:48:45,100
CAN EXPOSE ANOTHER DEVELOP TO

2477
01:48:45,100 --> 01:48:53,642
NETs AND INDUCE APOPS SO --

2478
01:48:53,642 --> 01:48:56,311
APOPTOSIS I'M NOT AWARE, IN

2479
01:48:56,311 --> 01:48:57,245
BYSTANDER CELLS, NOT NEUTROPHILS

2480
01:48:57,245 --> 01:48:58,446
BUT OTHER TYPES OF CELLS.

2481
01:48:58,446 --> 01:49:03,718
>> WE DO KNOW THEY CAN
INDUCE

2482
01:49:03,718 --> 01:49:05,387
ENDOTHELIAL CELL DEATH,

2483
01:49:05,387 --> 01:49:07,422
ESPECIALLY IN CONTEXT OF NETs

2484
01:49:07,422 --> 01:49:09,825
AUTOIMMUNE PATIENTS, GOOD AT

2485
01:49:09,825 --> 01:49:13,862
INDUCING, DO HAVE ABILITY TO

2486
01:49:13,862 --> 01:49:14,663
DAMAGE OTHER CELLS.

2487
01:49:14,663 --> 01:49:21,336
WE THINK THAT'S ONE OF THE

2488
01:49:21,336 --> 01:49:21,703
MECHANISMS.

2489
01:49:21,703 --> 01:49:23,405
>> NOW JUST AS YOU SAID
THAT A

2490
01:49:23,405 --> 01:49:27,175
QUESTION CAME IN WITH REGARD TO

2491
01:49:27,175 --> 01:49:28,476
NETs AND KIDNEY DISEASE.

2492
01:49:28,476 --> 01:49:32,080
AND OF COURSE ONE COULD THINK OF

2493
01:49:32,080 --> 01:49:35,417
THAT CONNECTION I GUESS THROUGH

2494
01:49:35,417 --> 01:49:36,484
THE GLOMERULUS.

2495
01:49:36,484 --> 01:49:40,055
ARE THERE VASCULAR BEDS THAT ARE

2496
01:49:40,055 --> 01:49:42,691
MORE OR LESS PRONE TO THE

2497
01:49:42,691 --> 01:49:45,460
EFFECTS OF NETosis AND WHAT IS

2498
01:49:45,460 --> 01:49:47,495
THE MECHANISM BY WHICH NETs

2499
01:49:47,495 --> 01:49:49,397
PERHAPS CAN INCREASE THE RISK OF

2500
01:49:49,397 --> 01:49:52,801
CHRONIC KIDNEY DISEASE IF THEY

2501
01:49:52,801 --> 01:49:52,968
DO?

2502
01:49:52,968 --> 01:49:55,437
>>  SO, THIS IS ACTUALLY
ONGOING

2503
01:49:55,437 --> 01:49:57,772
WORK IN OUR LAB.

2504
01:49:57,772 --> 01:50:01,209
MARIO, A POSTDOCTORAL FELLOW,

2505
01:50:01,209 --> 01:50:03,511
WORKING VERY -- FOCUSING ON

2506
01:50:03,511 --> 01:50:03,745
THAT.

2507
01:50:03,745 --> 01:50:05,213
WHAT WE'RE SEEING PRELIMINARY IS

2508
01:50:05,213 --> 01:50:12,187
WE'RE SEEING EVIDENCE OF

2509
01:50:12,187 --> 01:50:14,489
NEUTROPHILS INFILTRATING

2510
01:50:14,489 --> 01:50:15,156
PARTICULARLY GLOMERULAR REGIONS,

2511
01:50:15,156 --> 01:50:17,492
ONE MECHANISM THAT LEADS THE

2512
01:50:17,492 --> 01:50:23,431
FINDING BESIDES POTENTIAL EFFECT

2513
01:50:23,431 --> 01:50:27,636
ON VASCULAR HEALTH MAY STIMULATE

2514
01:50:27,636 --> 01:50:32,107
CELLS, A MECHANISM COULD LEAD TO

2515
01:50:32,107 --> 01:50:33,308
ENHANCED VASCULAR DAMAGE.

2516
01:50:33,308 --> 01:50:36,244
WORK ON NETs AND FIBROSIS,

2517
01:50:36,244 --> 01:50:37,712
CERTAIN CIRCUMSTANCES NETs CAN

2518
01:50:37,712 --> 01:50:39,047
BE STRONG INDUCERS OF FIBROSIS,

2519
01:50:39,047 --> 01:50:41,750
IN THE CONTEXT OF THE KIDNEY ONE

2520
01:50:41,750 --> 01:50:43,919
COULD SPECULATE THIS COULD BE

2521
01:50:43,919 --> 01:50:47,322
ANOTHER POTENTIAL MECHANISM.

2522
01:50:47,322 --> 01:50:48,323
WE'RE SEEING INFILTRATING NETs

2523
01:50:48,323 --> 01:50:52,360
IN BIOPSIES FROM LUPUS PATIENTS,

2524
01:50:52,360 --> 01:50:53,228
APPEARS TO BE HAPPENING IN VIVO

2525
01:50:53,228 --> 01:50:54,963
AS WELL.

2526
01:50:54,963 --> 01:50:56,731
>> YOU RAISED QUESTION OF

2527
01:50:56,731 --> 01:50:58,166
FIBROSIS, SO THEN THAT WOULD

2528
01:50:58,166 --> 01:51:01,002
LEAD SOME TO THINK ABOUT

2529
01:51:01,002 --> 01:51:03,405
SCLERODERMA AS AN AUTOIMMUNE

2530
01:51:03,405 --> 01:51:06,007
DISEASE WITH FIBROSIS BUT ONE

2531
01:51:06,007 --> 01:51:11,313
COULD THINK OF OTHER FORMS OF

2532
01:51:11,313 --> 01:51:11,947
FIBROSIS, INTERSTITIAL

2533
01:51:11,947 --> 01:51:22,457
PULMONARY,ER  IS A ROWS --  --

2534
01:51:23,491 --> 01:51:25,994
CIRRHOSIS OF THE LIVER.

2535
01:51:25,994 --> 01:51:30,065
>> THERE HAVE BEEN MAYBE
MORE

2536
01:51:30,065 --> 01:51:33,568
INDIRECT BUT I'VE SEEN THINGS SO

2537
01:51:33,568 --> 01:51:36,004
THERE IS SOME ON NEUTROPHILS IN

2538
01:51:36,004 --> 01:51:38,039
THE LIVER AND ACTUALLY TISSUE

2539
01:51:38,039 --> 01:51:38,606
RESIDENT NEUTROPHILS IN THE

2540
01:51:38,606 --> 01:51:40,842
LIVER, ONE OF THE ONES YOU CAN

2541
01:51:40,842 --> 01:51:47,382
FIND NEUTROPHILS EVEN IN ABSENCE

2542
01:51:47,382 --> 01:51:49,451
OF INFECTION, SO CERTAINLY NOT

2543
01:51:49,451 --> 01:51:52,120
ONLY RELATED TO LUPUS AND LUNGS,

2544
01:51:52,120 --> 01:51:54,823
BUT MORE WORK IS NEEDED

2545
01:51:54,823 --> 01:51:56,124
REGARDING SPECIFICITY AND

2546
01:51:56,124 --> 01:51:57,926
PATHWAYS THAT ARE TRIGGERED BY

2547
01:51:57,926 --> 01:51:58,994
THESE NETs.

2548
01:51:58,994 --> 01:52:00,161
>> ALL RIGHT.

2549
01:52:00,161 --> 01:52:03,465
NOW ANOTHER OF THE EVOLUTIONARY

2550
01:52:03,465 --> 01:52:05,266
BIOLOGISTS BACK WITH US, WANTING

2551
01:52:05,266 --> 01:52:07,969
TO KNOW THIS MAY HAVE IN SOME

2552
01:52:07,969 --> 01:52:09,871
WAY BEEN COVERED BY ARTURO BUT

2553
01:52:09,871 --> 01:52:11,873
I'LL POSE THE QUESTION.

2554
01:52:11,873 --> 01:52:16,478
THAT IS ARE THERE PLANT CELLS

2555
01:52:16,478 --> 01:52:18,546
THAT CAN UNDERGO NETosis?

2556
01:52:18,546 --> 01:52:21,216
>> YES. 

2557
01:52:21,216 --> 01:52:22,817
THERE'S A BEAUTIFUL WORK OF --

2558
01:52:22,817 --> 01:52:25,553
I'M BLOCKING THE NAME, SORRY.

2559
01:52:25,553 --> 01:52:27,155
SHE PASSED AWAY RECENTLY

2560
01:52:27,155 --> 01:52:28,289
UNFORTUNATELY, SHE SHOWED THERE

2561
01:52:28,289 --> 01:52:32,027
ARE SPECIFIC CELLS IN THE ROOTS

2562
01:52:32,027 --> 01:52:34,062
OF PLANTS THAT RE-FORM THEIR

2563
01:52:34,062 --> 01:52:39,000
CHROMATIN AND EXPEL INTO THE

2564
01:52:39,000 --> 01:52:39,367
SOIL.

2565
01:52:39,367 --> 01:52:42,070
WHAT IS INTERESTING IS THAT SHE

2566
01:52:42,070 --> 01:52:44,739
DID A VERY SIMPLE EXPERIMENT OF

2567
01:52:44,739 --> 01:52:47,442
INFECTING A PLANT WITH A PLANT

2568
01:52:47,442 --> 01:52:49,344
FUNGAL PATHOGEN AND SHOWED IF

2569
01:52:49,344 --> 01:52:51,946
SHE DESTROYED LET'S CALL THEM

2570
01:52:51,946 --> 01:52:54,149
PLANT NETs, PLANT BECAME FAR

2571
01:52:54,149 --> 01:52:57,752
MORE SUSCEPTIBLE TO THE FUNGAL

2572
01:52:57,752 --> 01:52:58,186
INFECTION.

2573
01:52:58,186 --> 01:52:59,854
SO IT IS MODIFIED CHROMATIN THAT

2574
01:52:59,854 --> 01:53:01,523
HAS THE FUNCTION OF IMMUNE

2575
01:53:01,523 --> 01:53:05,260
DEFENSE, OF COURSE IT DOESN'T

2576
01:53:05,260 --> 01:53:06,194
HAVE -- I DON'T THINK ANYBODY

2577
01:53:06,194 --> 01:53:10,598
HAS WORKED ON THE COMPOSITION OF

2578
01:53:10,598 --> 01:53:13,001
THESE PLANT NETs TO SEE THE

2579
01:53:13,001 --> 01:53:15,703
EQUIVALENT OF THE PEROXIDASE, ET

2580
01:53:15,703 --> 01:53:19,374
CETERA, AS FAR AS I KNOW, NO

2581
01:53:19,374 --> 01:53:23,445
WORK ON THE FORMATION OF NETs

2582
01:53:23,445 --> 01:53:24,946
PLANT FORMATION.

2583
01:53:24,946 --> 01:53:25,947
>> THERE ARE STUDIES

2584
01:53:25,947 --> 01:53:27,415
DEMONSTRATING -- THEY GO BACK A

2585
01:53:27,415 --> 01:53:29,617
LOT FURTHER IN PLANTS WITH

2586
01:53:29,617 --> 01:53:32,620
REGARD TO INNATE IMMUNITY, AND

2587
01:53:32,620 --> 01:53:34,022
VARIOUS MECHANISMS OF INNATE

2588
01:53:34,022 --> 01:53:36,324
IMMUNITY BUT I GUESS PLANTS

2589
01:53:36,324 --> 01:53:40,495
DON'T REALLY HAVE THE EQUIVALENT

2590
01:53:40,495 --> 01:53:42,797
OF NEUTROPHILS, FOR EXAMPLE,

2591
01:53:42,797 --> 01:53:46,067
THERE ISN'T A, I DON'T THINK,

2592
01:53:46,067 --> 01:53:49,404
BUT I'M ASKING, THEY DON'T HAVE

2593
01:53:49,404 --> 01:53:50,839
AN EQUIVALENT OF A NEUTROPHIL

2594
01:53:50,839 --> 01:53:53,575
BUT IT MAY BE THAT THIS IS AN

2595
01:53:53,575 --> 01:53:54,542
EXAMPLE OF CONVERGENT EVOLUTION

2596
01:53:54,542 --> 01:54:00,849
OR WHAT HAVE YOU IN WHICH THIS

2597
01:54:00,849 --> 01:54:03,751
MECHANISM IS VERY IMPORTANT FOR

2598
01:54:03,751 --> 01:54:05,286
ANTIMICROBIAL DEFENSE, AND THERE

2599
01:54:05,286 --> 01:54:08,823
HAVE BEEN MORE THAN ONE PATHWAY

2600
01:54:08,823 --> 01:54:14,863
FOR IT TO EVOLVE, I GUESS ONE

2601
01:54:14,863 --> 01:54:16,297
MIGHT THINK.

2602
01:54:16,297 --> 01:54:16,998
THAT'S FASCINATING.

2603
01:54:16,998 --> 01:54:22,270
WE'RE CLOSE TO THE END OF OUR

2604
01:54:22,270 --> 01:54:22,871
SESSION.

2605
01:54:22,871 --> 01:54:25,306
I GUESS I MIGHT START WRAPPING

2606
01:54:25,306 --> 01:54:27,742
THINGS UP BY ASKING THE TWO OF

2607
01:54:27,742 --> 01:54:32,413
YOU WHAT YOU THINK ARE THE MOST

2608
01:54:32,413 --> 01:54:35,416
IMPORTANT UNRESOLVED ISSUES IN

2609
01:54:35,416 --> 01:54:36,918
THE WORLD OF NETosis, AT

2610
01:54:36,918 --> 01:54:40,388
LEAST, YOU KNOW, FROM YOUR

2611
01:54:40,388 --> 01:54:40,955
PERSPECTIVE.

2612
01:54:40,955 --> 01:54:43,558
ARTURO, DOUGH WANT TO START OUT

2613
01:54:43,558 --> 01:54:47,428
MAYBE WITH REGARD TO THE BASIC

2614
01:54:47,428 --> 01:54:50,832
SCIENCE OF NETosis AND WHAT

2615
01:54:50,832 --> 01:54:54,202
ARE THE BIG UNRESOLVED QUESTIONS

2616
01:54:54,202 --> 01:54:58,640
THAT YOUNG INVESTIGATORS MIGHT

2617
01:54:58,640 --> 01:54:59,073
WANT TO ATTACK?

2618
01:54:59,073 --> 01:55:00,141
>>  WELL, I THINK
UNDERSTANDING

2619
01:55:00,141 --> 01:55:03,478
MECHANISM WHAT IS IS BEING

2620
01:55:03,478 --> 01:55:04,879
OXIDIZED IN NET FORMATION IS A

2621
01:55:04,879 --> 01:55:07,081
GOOD QUESTION, HOW IS CHROMATIN

2622
01:55:07,081 --> 01:55:09,984
MODIFIED IN ORDER TO ALLOW THE

2623
01:55:09,984 --> 01:55:11,019
UNWRAPPING AND FORMATION OF

2624
01:55:11,019 --> 01:55:11,586
NETs.

2625
01:55:11,586 --> 01:55:12,820
THAT'S TWO QUESTIONS THAT COME

2626
01:55:12,820 --> 01:55:13,354
TO MIND.

2627
01:55:13,354 --> 01:55:15,557
I STILL THINK THAT THE

2628
01:55:15,557 --> 01:55:18,459
ANTIMICROBIAL ACTIVITY OF

2629
01:55:18,459 --> 01:55:19,260
HISTONES IS EXTREMELY

2630
01:55:19,260 --> 01:55:19,627
INTERESTING.

2631
01:55:19,627 --> 01:55:22,564
AND I WOULD START A LAB ON THAT

2632
01:55:22,564 --> 01:55:26,067
IF I WERE STARTING A LAB AGAIN.

2633
01:55:26,067 --> 01:55:29,938
>> WELL, THOSE ARE REALLY
ALL

2634
01:55:29,938 --> 01:55:30,405
FASCINATING QUESTIONS.

2635
01:55:30,405 --> 01:55:33,741
MARIANA, WHAT ABOUT FROM YOUR

2636
01:55:33,741 --> 01:55:35,443
VANTAGE POINT?

2637
01:55:35,443 --> 01:55:39,414
>>  I THINK ANOTHER
QUESTION I

2638
01:55:39,414 --> 01:55:44,352
HAVE THIS CONCEPT OF NEUTROPHIL

2639
01:55:44,352 --> 01:55:44,919
HETEROGENEITY.

2640
01:55:44,919 --> 01:55:46,721
WHEN YOU STIMULATE, ONLY A

2641
01:55:46,721 --> 01:55:47,655
CERTAIN PERCENTAGE UNDERGO

2642
01:55:47,655 --> 01:55:49,757
NETs, THE REST DON'T, RIGHT?

2643
01:55:49,757 --> 01:55:50,858
WHY IS THAT?

2644
01:55:50,858 --> 01:55:54,095
AND WE REALLY HAVE JUST STARTED

2645
01:55:54,095 --> 01:55:54,996
TO UNDERSTAND THAT NOT ALL

2646
01:55:54,996 --> 01:55:58,800
NEUTROPHILS ARE THE SAME.

2647
01:55:58,800 --> 01:56:00,969
I THINK THAT POTENTIALLY WE WANT

2648
01:56:00,969 --> 01:56:02,070
TO TARGET ABNORMAL NET FORMATION

2649
01:56:02,070 --> 01:56:11,679
WOULD BE SO USEFUL BECAUSE WE --

2650
01:56:11,679 --> 01:56:14,048
A MORE TARGETED APPROACH IS

2651
01:56:14,048 --> 01:56:15,717
NEEDED, TO ME THAT'S ONE OF THE

2652
01:56:15,717 --> 01:56:19,654
KEY QUESTIONS AND SOMETHING THAT

2653
01:56:19,654 --> 01:56:20,855
KEEPS ME UP WHEN I THINK ABOUT

2654
01:56:20,855 --> 01:56:24,292
INTERESTING THINGS TO DO IN THE

2655
01:56:24,292 --> 01:56:24,525
FUTURE.

2656
01:56:24,525 --> 01:56:32,000
I HOPE OTHERS WILL KEEP WORKING

2657
01:56:32,000 --> 01:56:33,835
ON MOVING FORWARD.

2658
01:56:33,835 --> 01:56:35,436
>> IT'S BEEN A FASCINATING

2659
01:56:35,436 --> 01:56:35,670
SESSION.

2660
01:56:35,670 --> 01:56:37,839
I THINK AT THIS POINT WE CAN

2661
01:56:37,839 --> 01:56:40,008
CALL IT TO A CLOSE.

2662
01:56:40,008 --> 01:56:44,512
THANK YOU SO, SO MUCH FOR REALLY

2663
01:56:44,512 --> 01:56:47,415
A WONDERFUL, WONDERFUL SET OF

2664
01:56:47,415 --> 01:56:49,450
PRESENTATIONS, AND A VERY

2665
01:56:49,450 --> 01:56:50,451
ENGAGING DISCUSSION HERE IN THE

2666
01:56:50,451 --> 01:56:54,255
LAST HALF HOUR OR SO.

2667
01:56:54,255 --> 01:56:56,257
AGAIN, WE REALLY APPRECIATE IT.

2668
01:56:56,257 --> 01:56:58,993
GOOD LUCK TO BOTH OF YOU ON

2669
01:56:58,993 --> 01:57:02,864
SOLVING THESE QUESTIONS THAT YOU

2670
01:57:02,864 --> 01:57:04,799
HAVE YOU NOW POSED AND WE'LL

2671
01:57:04,799 --> 01:57:06,901
LOOK FORWARD TO SEEING YOU BACK

2672
01:57:06,901 --> 01:57:10,705
IN THE NOT TOO DISTANT FUTURE

2673
01:57:10,705 --> 01:57:11,873
WITH PERHAPS THE NEXT

2674
01:57:11,873 --> 01:57:14,108
INSTALLMENT IN THE FIELD OF

2675
01:57:14,108 --> 01:57:14,342
NETosis.

2676
01:57:14,342 --> 01:57:18,746
SO THANK YOU VERY MUCH.

2677
01:57:18,746 --> 01:57:21,349
GOOD EVENING TO ARTURO AND GOOD

2678
01:57:21,349 --> 01:57:23,384
AFTERNOON TO MARIANA.

2679
01:57:23,384 --> 01:57:25,153
AND THANK EVERYONE IN OUR

2680
01:57:25,153 --> 01:57:27,455
AUDIENCE, ALL OVER THE WORLD,

2681
01:57:27,455 --> 01:57:29,724
FOR JOINING US TODAY.

2682
01:57:29,724 --> 01:57:31,426
ANYWAY, THANK YOU AGAIN AND SEE

2683
01:57:31,426 --> 01:57:33,127
YOU NEXT WEEK.

2684
01:57:33,127 --> 01:57:33,361
BYE-BYE.

2685
01:57:33,361 --> 01:57:35,163
>> THANKS FOR  INVITING
US.

2686
01:57:35,163 --> 01:57:45,163
[END OF PROGRAM]