I'M JACKIE MAYS, DIRECTOR OF THE FELLOWSHIP PROGRAM. OUR FELLOWSHIP PROGRAM WAS ESTABLISHED IN ITS CURRENT FORM IN 2010, AND WE CURRENTLY HAVE FIVE OUTSTANDING CLINICAL RESEARCH FELLOWS HERE AT THE NIDCR. I WANT TO WELCOME TODAY TO THE FIRST OF THE SERIES OF THE 2022 NIDCR CLINICAL FELLOWSHIP GRAND ROUNDS SERIES FOR THIS YEAR. WE'RE ABSOLUTELY THRILLED TO BE HOSTING THIS SERIES TODAY LIVE FROM LIPSETT AUDITORIUM, AT THE NIH CLINICAL CENTER. THIS GRAND ROUND SERIES WAS INITIATED IN 2014, BY DR. JANICE LEE, NAMED AFTER OUR CLINICAL FELLOWSHIP PROGRAM, IT'S THE FIRST NIH GRAND ROUND SERIES THAT'S DEDICATED TO THE TRAINEES, THE PURPOSE OF OUR GRAND ROUNDS SERIES IS TO HAVE LEADING SCIENTISTS AND CLINICIANS ADDRESS ADVANCES IN CLINICAL TRANSLATIONAL AND BASIC RESEARCH, IN AREAS RELATED TO THE DENTAL, ORAL, CRANIOFACIAL COMPLEX, ALSO BONE METABOLISM. AND IMPORTANTLY TO GIVE OUR TRAINEES OPPORTUNITIES TO NETWORK WITH THESE EXCEPTIONAL SCIENTISTS. WE'VE HAD SPEAKERS OF ALL BACKGROUNDS, VIA VIDEOCAST SINCE THE INCEPTION SO THE ENTIRE MEDICAL AND DENTAL COMMUNITY CAN SHARE IN LISTENING TO THESE EXCEPTIONAL SPEAKERS. THIS QUARTERLY SERIES IS HELD THREE TIMES YEARLY ARE THE ANNUAL STATE AWARDS CEREMONY IN LIEU OF ONE OF THE GRAND ROUNDS. THE GRAND ROUNDS THEME HIGHLIGHTED WOMEN IN SCIENCE, THIS YEAR THE YEAR OF INNOVATION LOOKING FORWARD TO HIGHLIGHTING SCIENTIFIC AND MEDICAL ADVANCES. I HOPE THE TRADITION OF GRAND ROUNDS AND RECOGNIZING IMPORTANCE OF OUR FELLOWSHIP CONTINUES IN PERPETUITY. I'D LIKE TO WELCOME JANICE LEE TO THE PODIUM TO INTRODUCE OUR SPEAKER. >> THANK YOU VERY MUCH, DR. MAYS. AGAIN, WELCOME TO THE NIDCR CLINICAL RESEARCH FELLOWSHIP GRAND ROUNDS, THE YEAR OF INNOVATION. DR. SOUSA AND DR. TABAK WOULD HAVE INTRODUCED THE SPEAKER, DR. SOUSA IS GIVING A PRESENTATION, DR. TABAK HUSTLING TO THE WHITE HOUSE SO YOU'RE STUCK WITH ME. IT'S A PLEASURE TO INTRODUCE OUR SPEAKER, DR. JENNIFER WEBSTER-CYRIAQUE, OUR DEPUTY DIRECTOR FOR THE NIDCR, JOINED US IN DECEMBER 2021. BEFORE WE GOT SWEPT UP IN ALL HER DUTIES WE DECIDED TO NAB HER FOR GRAND ROUNDS. AS DR. MAYS SUGGESTED, THE GRAND ROUNDS ALLOWS US TO HAVE AN OPPORTUNITY FOR OUR CLINICAL RESEARCH FELLOWS TO MEET WITH LUMINARIES IN THEIR FIELDS. BECAUSE OF HER TRACK RECORD IN MENTORING, RESEARCHING, CLINICAL EXCELLENCE, IT'S A PLEASURE TO HAVE DR. WEBSTER-CYRIAQUE PARTICIPATE IN THIS WAY. SHE RECEIVED HER UNDERGRADUATE AND DENTAL DEGREE FROM SUNY IN BUFFALO AND Ph.D. FROM UNIVERSITY OF NORTH CAROLINA, CHAPEL HILL IN 1998, FACULTY MEMBER AT THE UNIVERSITY OF NORTH CAROLINA, SCHOOLS OF DENTISTRY AND MEDICINE, FOR MORE THAN TWO DECADES AND THEN ROSE THROUGH THE RANKS TO FULL PROFESSOR TENURED. WHILE THERE CONDUCTED BASIC AND CLINICAL RESEARCH TO DETERMINE THE ETIOLOGY FOR SALIVARY GLAND DISEASE IN PATIENTS WITH HIV, CHARACTERIZED ORAL HPV, ASSESSED ORAL MICROBIOME, IMPLICATIONS FOR CANCER-CAUSING VIRUS AND OUTCOME TO HIV, AND HAS LED MANY LEADERSHIP ROLES INCLUDING CHAIR OF THE ORAL HIV/AIDS RESEARCH ALLIANCE, RESEARCH DIRECTOR AT THE NATIONAL DENTAL ASSOCIATION FOUNDATION, AND HAS HAD SEVERAL ROLES WITHIN THE AMERICAN ASSOCIATION FOR DENTAL, ORAL, CRANIOFACIAL RESEARCH AND SISTER INTERNATIONAL ORGANIZATION. I'M THRILLED TO HAVE DR. WEBSTER-CYRIAQUE DISCUSS HER WORK AND THE CURRENT STATE OF ORAL HEALTH IN THE SETTING OF HPV AND HIV. MANY OF YOU RECOGNIZE A HUMAN PAPILLOMAVIRUS HPV CAN CAUSE SERIOUS HEALTH OUTCOMES AND PROBLEMS INCLUDING OROPHARYNGEAL CANCER, THE MOST COMMON SEXUALLY TRANSMITTED INFECTION IN THE U.S. AND ABOUT 10% OF MEN AND NEARLY 4% OF WOMEN HAVE HPV. BECAUSE HPV IS THOUGHT TO CAUSE 70% OF OROPHARYNGEAL CANCERS IN THE U.S., AND WE CONTINUE TO SEE THE IMPACT OF HEALTH DISPARITIES OF HIV ON ORAL HEALTH, WE CAN SEE WHY THESE AREAS ARE IMPORTANT AND ARE SERIOUS CONSIDERATIONS FOR RESEARCH AND CLINICAL CARE. WITHOUT FURTHER ADO I INDICT DR. WEBSTER-CYRIAQUE TO THE PODIUM. [APPLAUSE] >> THANK YOU SO MUCH, DR. MAYS AND DR. LEE, FOR THIS AWESOME INTRODUCTION AND THIS GREAT OPPORTUNITY TO BE HERE. I I CAN'T TELL YOU HOW EXCITED I AM TO BE HERE AND TO JUST SHARE WITH YOU GUYS A LITTLE BIT OF WHAT WE'VE LEARNED OVER THE LAST FEW YEARS. SO WITHOUT FURTHER ADO ... WE'RE GOING TO TALK ABOUT WHAT HAS HAPPENED OVER TIME. THIS IS THE ADVANTAGE, AS YOU GUYS COLLEAGUE CLINICAL FELLOWS, I THOUGHT I WOULD REALLY TAKE THIS ENTIRE TALK FROM A CLINICAL STANDPOINT AND AS DR. LEE JUST SHARED WITH YOU, I STARTED MY PhD in the '90s, EARLY '90s, JUST GRADUATED FROM DENTAL SCHOOL AT THAT TIME AND WE WERE AT THE HIV EPIDEMIC. WE REALLY DID NOT HAVE MANY TOOLS OR THERAPIES WITH REGARD TO HIV. AT THAT POINT THERE WAS SIGNIFICANT ORAL DISEASE WE SAW IN THE ORAL CAVITY INCLUDING THE DISEASE CAUSED BY EPSTEIN-BARR VIRUS AND KAPOSI SAR SARCOMA. WE BEGAN TO SEE LESS WITH MEDICATION BUT STARTED TO SEE A SALIVARY GLAND DISEASE CROP UP AND TONS OF WARTS ACROSS THE FIELD. WE ALSO BEGAN TO SEE THROUGHOUT THE CLINIC POPULATION AS WE MOVED INTO THE 2000s AN INCREASE IN VIRUS-DRIVEN HEAD AND NECK CANCERS, WE'RE GETTING MORE HPV-ASSOCIATED DISEASE. AND OVER TIME WE'VE BEGUN TO DEVELOP A SIGNIFICANT RECOGNITION OF POLYMICROBIAL CONDITIONS, GIVEN THE ADVENT OF TECHNOLOGY AND OUR UNDERSTANDING OF THE DIFFERENT MICROBIOMES. AND THROUGHOUT ALL OF THIS TIME, WHEN WE THINK ABOUT WHAT'S HAPPENING FOR THIS VULNERABLE POPULATION, DENTAL CARE REMAINS A PROBLEM FOR PEOPLE LIVING WITH HIV. IT'S THE TOP -- IT'S WITHIN THE TOP THREE UNMET NEEDS, RIGHT? I'M TALKING ABOUT HAVING ENOUGH FOOD TO EAT, HAVING TRANSPORTATION, A ROOF OVER YOUR HEAD, NOT HAVING DENTAL CARE IS RIGHT UP THERE, OKAY? AND THERE ARE ACTUALLY MORE PEOPLE THAT REPORT UNMET DENTAL NEEDS THAN UNMET MEDICAL NEEDS IN THE HIV COMMUNITY. SO IT'S BEEN MY PRIVILEGE OVER THE LAST COUPLE DECADES AS A HOSPITAL DENTIST TO TREAT THIS GROUP OF INDIVIDUALS TOGETHER WITH OTHER PEOPLE MEDICALLY COMPROMISED, MUCH LIKE YOU SEE HERE, PEOPLE WHO HAVE CANCER AND PEOPLE WHO ARE HEMOPHILIAC, SICKLE CELL, WHAT HAVE YOU. AMONG THESE PEOPLE YOU SEE PEOPLE LIVING WITH HIV. NOW, WE KNOW THE DENTAL DISEASE IS A PROBLEM IN GENERAL, AND IN THE UNITED STATES CLOSE TO HALF OF ADULTS IN THE UNITED STATES HAVE PERIODONTAL DISEASE AND WE SEE DISPARITIES WITH HISPANICS AND AFRICAN AMERICANS HAVING HIGHER LEVELS. BUT WHEN WE LOOK AT HIV, THE PREVALENCE OF GUM DISEASE GOES UP TO 75%. SO DENTAL DISEASE IS A MAJOR PROBLEM IN THE CONTEXT OF HIV. AND SO WE WANTED TO THEN -- WE DID A LARGER STUDY ON ABOUT 200 PEOPLE. THIS WAS ACTUALLY A HRSA-FUNDED STUDY THAT ALLOWED US TO PROVIDE CARE FOR PEOPLE WITH HIV. AND WITHIN A SUBSET OF THAT GROUP WE DECIDED TO ASK QUESTIONS ABOUT THE RELATIONSHIP BETWEEN THE ORAL MICROBIOME AND DISEASE. THEY WERE SEEN FOR 24 MONTHS AT LEAST EVERY SIX MONTHS, COMPREHENSIVE EXAM AND TREATMENT PLAN, INTERVIEWED, DEBRIDEMENT, ORAL HEALTH INSTRUCTION, LOOK AT PERIOCHARTING, ORAL HYGIENE INSTRUCTIONS, AND THEN WE COLLECTED SAMPLES FROM THEM AND SAW THEM EVERY SIX MONTHS, TOOK SAMPLES EVERY 12 MONTHS. BUT HERE THE ORAL HEALTH CARE WAS THE INTERVENTION. MEAN AGE 45, MOSTLY MALE, AND ABOUT 37% WERE SMOKERS. AND LARGE NUMBER OF AFRICAN AMERICANS. AND WHAT'S INTERESTING IS THAT PERIODONTAL HEALTH AS MEASURED BY LOWER LEVELS OF INFLAMMATION, LOWER BLEEDING ON PROBING, WAS ASSOCIATED WITH DIFFERENCE IN THE MICROBIOME, SO THE PEOPLE WITH LOWER BLEEDING ON PROBING HAD MORE NICERIA, LESS PREVOTELLA, WHICH IS A PATHOGEN, COMPARED TO PEOPLE ARE HIGH BLEEDING ON PROBING. WE SEE WHEN PEOPLE ARE ON ANTIRETROVIRAL MEDICATION, YOU SEE A HEALTHIER MICROBIOME WITH MORE STREP, MORE LACTOBACILLUS, LOOKING AT KEY PATHOGENS, LOWER LEVELS IN TERMS OF ABUNDANCE OF THIS ORGANISM THAN THOSE PEOPLE NOT ON MEDS. WHERE YOU ARE AND WHAT'S HAPPENING IN TERMS OF MEDICATION MAKES A DIFFERENCE. WHAT ABOUT RACIAL DIFFERENCE? JUST IN PERIODONTAL DISEASE IN GENERAL, AFRICAN AMERICANS AND HISPANICS HAVE HIGHER LEVELS, WHAT WAS INTERESTING WAS AT BASELINE WE ACTUALLY SEE A DIFFERENCE IN THE MICROBIOME HERE WHERE YOU HAVE A MORE PATHOGENIC BIOMARKER WITH POPPHHYR MONOS, DIFFERENCE IN WHITE AND BLACK GROUP BUT WITH CARE OVER TIME THEY START TO LOOK THE SAME IN TERMS OF ALPHA DIVERSITY. WE LOOK AT THIS IN THE CONTEXT OF THEIR CD4 COUNT, BASELINE IS IN THE BLUE, SO FOR WHITE GROUP AND BLACK GROUP YOU HAVE LOWER CD4 COUNTS AT BASELINE BUT THE BLACK GROUP WAS LOWER THAN THE WHITE GROUP. WITH DENTAL CARE THE CD 4 COUNTS RISE AND ARE SIMILAR, SO DENTAL CARE IS HELPING TO CLOSE THE GAP. WITH DENTAL CARE WE SEE INCREASING CD4 COUNT, AND THAT IS VERY CLOSELY ASSOCIATED WITH DECREASED BLEEDING ON PROBING, DECREASED ORAL INFLAMMATION, AND WE CAN SEE CLEARLY THIS NEGATIVE ASSOCIATION. AND WHAT'S INTERESTING IS WHEN YOU ACTUALLY LOOK AT THE MICROBIOME OF A PERSON WHO HAS A LOW CD4 COUNT YOU SEE MUCH MORE BACTERIAL RICHNESS HERE COMPARED TO THE CD4 HIGH GROUP. NOW, WHAT'S COOL IS, TO ME, NOT ONLY ARE WE SEEING -- WHEN WE TELL PEOPLE WHAT THEY NEED TO DO AT HOME AND ARE GETTING TREATMENT AND COMPREHENSIVE DENTAL CARE THEY NEED NOT ONLY DO WE SEE THE CYTOKINES THAT ARE INFLAMMATORY MEDIATORS CLOSELY ASSOCIATED WITH SYSTEMIC INFLAMMATION IN THE CONTEXT OF HIV, HERE WE'RE LOOKING IN THE SALIVA AT BASELINE, 12 MONTHS WE SEE IT GO DOWN, 24 MONTHS WAY DOWN, COMPARED TO HEALTHY CONTROL, OKAY? NOT ONLY IS THAT GOING DOWN IN THE SALIVA, IT'S GOING DOWN IN THE BLOOD. SO WHAT'S HAPPENING IN THE MOUTH HERE IS MAKING A DIFFERENCE. WE SEE VERY SIMILAR STORY WITH IL-6 THAT GOES DOWN. SAME WITH D-DIMER AND OTHER CYTOKINES. WE STRATIFIED OUR GROUP BASED ON MEDICATION STATUS, PEOPLE WHO WERE NOT ON A.R.T. YET, NEWLY DIAGNOSED GROUP, ON MEDICINE LESS THAN A YEAR, ON MEDICATION FOR MORE THAN A YEAR, AND THAT MEDICINE WAS WORKING. THEY WERE SUPPRESSED. THEY HAD UNDETECTABLE VIRAL LOADS AND PEOPLE NOT SUPPRESSED, WHAT I'LL TELL YOU AND DON'T HAVE TIME TO SHOW YOU, THOSE PEOPLE ON MEDICATION NOT SUPPRESSED ALL WENT TO SUPPRESSION AT THE END OF THE STUDY. BUT THE OTHER ONE MAJOR PROBLEM FOR PEOPLE WHO HAVE HIV AND ARE ON ANTIRETROVIRAL THERAPY, SOMETIMES THEY STILL HAVE PROBLEMS WITH INFLAMMATION AND THIS ALLOWS METABOLIC SYNDROMES TO MOVE FORWARD, THIS ALLOWS OPPORTUNISTIC INFECTIONS TO MANIFEST. AND THIS ALLOWS A MAINTENANCE OF LOWER CD4 COUNTS THAT HAVEN'T BEEN OPTIMIZED. WE CAN SEE THAT HERE WHERE WE LOOK AT IL-6 IN LONG-TERM SUPPRESSED GROUP AND SEE HOW SIGH IL-6 LEVELS ARE, WHEN WE PROVIDE DENTAL INTERVENTION IT DROPS RIGHT DOWN. WHEN WE LOOK AT CD4 COUNTS ACROSS THIS GROUP, AND ACTUALLY ALL THE GROUPS WE SAW IT GO UP BUT WE'RE NOT GOING TO COUNT NO ART OR SHORT TERM BECAUSE THAT COULD BE MEDICINE BUT ON ART OVER A YEAR WE SEE THE DENTAL INTERVENTION WAS ASSOCIATED WITH MEAN INCREASE OF ABOUT 168 INCREASE IN CD4 T CELL COUNT. SO, WHAT'S HAPPENING IN THE MOUTH IN A SIMPLE LOW COST INTERVENTION CAN HAVE PRETTY GOOD HIGH IMPACT WITH REGARD TO THE OVERALL SYSTEMIC CONDITION OF HIV. AND JUST SOME MORE INFORMATION HERE TO DEMONSTRATE THAT WE SAW LEVELS OF IP-10 DECREASE, ALSO HERE LOOKING IN THE BLOOD AT SIX MONTHS, AND LOOKING AT SYSTEMIC PRO-INFLAMMATORY CYTOKINE LEVELS AND WE CAN SIGH SIGNIFICANT CHANGES FROM BASELINE TO SIX MONTHS, IT'S FUZZY BUT YOU CAN SEE KNOWN PRO-INFLAMMATORY MEDIATORS, IL-1, IL-17, IL-15. SO, DENTAL CARE MAKES A DIFFERENCE, NOT JUST TO THE MOUTH BUT TO THE BODY. SO, WHAT ELSE DO WE SEE IN THE ORAL CAVITY OF PEOPLE WITH HIV AND WHY? THIS IS THE QUESTION WE'RE ASKING. ONE THING WE SAW WHEN WE'VE LOOKED IN THE COURSE OF ONE OF OUR AIDS CLINICAL TRIALS GROUP STUDIES, SO WE, THE ORAL HIV/AIDS RESEARCH ALLIANCE, DID SEVERAL STUDIES IN THE CONTEXT OF THE NIAID AIDS CLINICAL TRIALS GROUP TO LOOK AT FREQUENCY OF HIV-RELATED ORAL LESIONS, U.S. SITES, WE HAD FIVE U.S. SITES AND ONE NON-U.S. SITE, THAT WAS IN HAITI, OKAY? AND THIS WAS JUST A COUPLE YEARS AGO. NOW, PEOPLE THINK, WELL, PEOPLE LIVING WITH HIV, THEY ARE ON MEDICINES, THERE'S NO MORE ORAL DISEASE. THAT'S NOT RIGHT. THAT'S WRONG. OKAY? BECAUSE WE'RE STILL SEEING HIGH LEVELS OF CANDIDIASIS, 50%, HAIRY LEUKOPLAKIA, HIGHER LEVELS IN THE NON-U.S., STILL SEEING KS AT THE NON-U.S. SITES, EXAMPLE OF ORAL KS. EXPLOSION OF ORAL WARTS LIKE I SHARED WITH YOU WHEN PEOPLE FIRST GOT ON PROTEASE INHIBITORS, THOSE HAVEN'T GONE AWAY ALL THE WAY. WE ALSO SEE THIS PAROTID GLAND ENLARGEMENT IN 10%, SALIVARY GLAND DISEASE. ORAL DISEASE REMAINS A PROBLEM AND CHALLENGE FOR PEOPLE LIVING WITH HIV. SO WE'RE GOING TO TALK A LITTLE BIT ABOUT THESE VIRUSES THAT DRIVE CANCERS IN THE MOUTH AND WE'LL START OUT WITH HIV SALIVARY GLAND DISEASE, PATH TO FIGURE OUT WHAT WAS GOING ON THERE. WE'LL TALK ABOUT HPV AND END UP WITH EPSTEIN BARR VIRUS. YOU CAN SEE HERE THIS ENLARGEMENT OF SALIVARY GLAND. A PROBLEM BECAUSE THE GLANDS ARE NOT WORKING, PEOPLE HAVE MORE DENTAL DISEASE. THE OTHER THING IS THIS IS AN EXTERNAL SIGN THAT A PERSON IS LIVING WITH HIV AND THIS CAN BE STIGMA ASSOCIATED WITH IT. THE OTHER MAJOR THING, A PERCENTAGE OF THESE PEOPLE WILL GO ON TO DEVELOP CANCER IN THAT GLAND. SO WE SAW OVER TIME PRE-HAART AND POST-HAART, A MUCH LOWER PREVALENCE IN THE MID-90S AND WHEN PEOPLE GOT ON PROTEASE INHIBITORS SAW IT INCREASE 5%, AND AS WE GOT TOWARDS 2010 WEEP GOT UP TO 8% SO WE SEE INCREASE AND THE LAST STUDY, 2016, 10%. THIS IS INCREASED. ONE OF MY COLLABORATORS NOW AT NIAID SHOWED US WHEN THIS HAPPENS, WHEN PEOPLE ARE ON ANTIRETROVIRAL THERAPY BUT MAY STILL HAVE DETECTABLE HIV RNA, SO THINGS QUITE HAVEN'T SETTLED YET. CD4 COUNTS HAVEN'T GONE UP, MAYBE ANTIRETROVIRAL THERAPY IS NOT 100% EFFECTIVE YET. WHAT DO WE THINK IS HAPPENING? IF THIS IS A CANCER-CAUSING VIRUS POPPING UP THE SAME TIME WE'RE SEEING OTHER OPPORTUNISTIC INFECTIONS THIS IS LIKELY GOING TO BE INFECTIOUS AGENT AND BECAUSE IT CAUSED LYMPHOMA, THIS HAS TO BE A DNA TUMOR VIRUS. SO WHAT WE THINK, WHAT I THOUGHT, IT'S LIKELY SOMETHING HAPPENS THAT WE GET INFECTED WITH THIS AGENT WHEN WE'RE KIDS, FOR MOST INFECTIONS AS CHILDREN, THERE'S NO DISEASE, IT HANGS OUT, AS LONG AS WE HAVE A GOOD IMMUNE SYSTEM. BUT AS WE MOVE TOWARDS IMMUNE SUPPRESSION, YOU CAN HAVE A REACTIVATION OF THESE THINGS. AND THIS MAY LEAD TO THE DISEASE. SO, THIS LEADS US TO THE HYPOTHESIS THAT HIV SALIVARY GLAND DISEASE WILL BE CAUSED BY OPPORTUNISTIC DNA TUMOR VIRUS. AND SO FIRST I THOUGHT IT WAS A HERPES VIRUS. HERE IS THE LESSON, YOU GOT TO WATCH THE DATA AND NOT MAKE ASSUMPTIONS. WHY DID I THINK THAT? THIS FAMILY OF VIRUSES, SHED INTO THE SALIVA, THEY HAVE BEEN DETECTED IN THE SALIVARY GLAND, INCLUDE VIRUSES LIKE CMV, EBV, HSV KNOWN IMMUNE SUPPRESSION OPPORTUNISTS, A SUBSET EBV AND KAPPASHV, I KNEW I WAS GOING TO DISCOVER A HERPES VIRUS, WE DID NOT DETECT A HERPES VIRUS HERE, HOWEVER WHEN WE WENT BACK TO SOME REALLY OLD LITERATURE, LIKE FROM THE 1950s, THERE WERE PAPERS THAT DESCRIBED MICE BEING INFECTED WITH POLYOMA VIRUS WHO DEVELOPED ENLARGED CAROTID GLANDS, PERCENTAGE OF THESE MICE WOULD GO TO DEVELOP A MALIGNANCY. OTHER SALIVARY GLANDS POLYOMA SEQUENCES HAVE BEEN DEVELOPED, AND THE OTHER THING IS ANOTHER POLYOMA VIRUS, BK, HAS BEEN DETECTED IN GLANDULAR TUMORS LIKE KIDNEY TUMORS AND PROSTATE GLAND, WHY NOT SALIVARY? WE DETECTED A BAND HERE, SEQUENCED IT AND LO AND BEHOLD IT LOOKS LIKE A POLYOMA VIRUS. WE DO ADDITIONAL STUDIES TO SEE IF THIS IS TRUE. FIRST LET ME TELL YOU ABOUT BKV. PRE-HIPAA THIS WAS NAMED BY THE PERSON'S INITIALS, BK. THIS IS 1971. AND THE VIRUS WAS ISOLATED FROM THE URINE OF A PERSON GETTING A RENAL TRANSPLANT. RIGHT NOW MOST OF US ARE INFECTED WITH BK VIRUS. IT'S UBIQUITOUS IN THE HUMAN POPULATION. PRIMARY INFECTION OCCURS IN CHILDHOOD, A MAJOR PROBLEM FOR PEOPLE IMMUNE SUPPRESSED, CAN CAUSE HEMORRHAGIC CYSTITIS, ONE OF THE MAJOR REASONS WHY PEOPLE REJECT KIDNEY TRANSPLANT. OKAY? THIS VIRUS, LIKE ALL POLYOMA VIRUS, IT'S A SMALL VIRUS, EXPRESSES A T ANTIGEN THAT BINDS TO PROTEINS THAT PROMOTE CANCER. P53 AND RB. AND WHAT WE FOUND WAS WE COULD CONSISTENTLY DETECT BKV DNA THAT LINES UP WITH BKV, SOMEWHAT BUT NOT SO WELL WITH OTHER POLYOMA VIRUS, WASHINGTON YOURS, SB 40 AND WE CAN DETECT RNA AND THE PROTEIN HERE. WE'RE CONSISTENTLY DETECTING IT AT ALL THREE LEVELS. WHEN WE ACTUALLY LOOK IN THE SALIVA OF INDIVIDUALS WHO HAVE THIS, WE CAN SEE THAT IN HIV SALIVARY GLAND DISEASE WE DETECT MUCH HIGHER LEVELS OF BKV, IN PEOPLE WHO HAVE THIS DISEASE THAN PEOPLE WHO ARE HIV POSITIVE AND DON'T HAVE THE DISEASE, WHO LOOK MORE LIKE PEOPLE WHO ARE HIV NEGATIVE AND DON'T HAVE SALIVARY GLAND DISEASE. WE BUILT AN IN VITRO SYSTEM TO FIGURE OUT WHAT WAS GOING ON AND BRUCE BAUM PROVIDED SALIVARY GLAND CELLS AND WE GFP LABELED THE VIRUS AND COULD DEMONSTRATE THIS VIRUS COULD INFECT SALIVARY GLAND CELLS, YOU CAN SEE GFP-LABELED VIRUS GOING IN. AND AFTER A COUPLE DAYS IN CULTURE WE COULD SEE VARIANTS COMING OUT, OKAY? THIS IS ELECTRON MICROSCOPY. WE CAN SEE THE VIRAL INFECTION SIGNIFICANTLY CHANGES THE MORPHOLOGY OF THESE CELLS, BUT THE OTHER THING THAT IT DID WHEN WE LOOK AT AMYLASE RNA WHICH IS AN IMPORTANT ENZYME MADE BY THE GLAND WHEN YOU INFECT, WITH THE VIRUS, YOU CAN SEE MUCH LOWER LEVELS OF THIS IMPORTANT ENZYME BEING MADE, OKAY? AND WE SEE SIGNIFICANT INCREASE IN PROLIFERATION WITH INFECTION. WELL, NOW WE DID A WHOLE BUNCH OF OTHER STUFF WHICH I WON'T BORE YOU WITH. AND WE'RE ABLE TO CHARACTERIZE THESE VIRUSES AND DEMONSTRATE THAT THEIR TROPISM, FOR THE MOUTH, SALIVARY GLAND, WAS DIFFERENT THAN TROPISM FOR THE KIDNEY. AND WE THEN -- NOW WE'VE HAD THE SYSTEM, WE DECIDED TO TEST SOME DRUGS SO WE TESTED CIDOFIV, LEFLUNIMIDE AND CIPRO, AN ANTIBACTERIAL. WHY? THE HELICASE LOOK LIKE THE BACTERIAL ONE. SO WE SCREENED THESE AND DECIDED ON CIPRO WORKED PRETTY WELL, AS WE COMPARE NO DRUG TO CIPRO TREATMENT. AND THESE ARE SEVERAL DIFFERENT BKVs THAT WERE ISOLATED FROM PATIENTS. SO WE DID A RANDOMIZED CONTROLLED TRIAL, SMALL ONE. AND WE CAN SEE TREATMENT WITH CIPROFLOXACIN ACTUALLY HAD LESS PEOPLE WHO WERE BKV POSITIVE, AND MORE BKV NEGATIVE WITH THOSE PEOPLE WHO HAD CIPROFLOXACIN COMPARED TO THE PLACEBO. SO, WE MOVED TO A STATE WHERE WE HAD NO IDEA WHAT WAS CAUSING THIS CLINICAL PROBLEM, AND SOME OF OUR COLLEAGUES, SURGEONS, WERE REMOVING PEOPLE'S GLANDS, OKAY? AND NOW WE HAVE A POTENTIAL TREATMENT. SO THAT'S THE BEAUTY OF MAKING THE CONNECTION, MOVING FROM THE CLINIC TO THE LAB AND BACK. SO HERE WE DETECTED A POTENTIAL ETIOLOGIC AGENT, DEVELOPED SALIVARY GLAND SYSTEM, WE CHARACTERIZED THE GENOMES AND PATHOGENESIS OF VARIANTS AND ACTUALLY COULD SEE THERE WERE SPECIFIC THINGS THAT OCCURRED WITHIN THE GENOMES OF THOSE PATHOGENS THAT COME OUT OF A SALIVARY GLAND OR FROM THE SALIVA THAT ARE DISTINCT FROM THOSE THAT ARE MORE KIDNEY TROPIC, OKAY? WE ACTUALLY DETECTED A LONG NON-CODING RNA, MALAT1, INDICATES P5 2 DEREGULATION AND NOW THIS LONG CODING RNA IS FAMOUS BECAUSE IT'S ASSOCIATED IN A NUMBER OF MALIGNANCIES. SUSIE FROM OUR GROUP DEVELOPED AN AGENT-BASED IN SILICO MODEL. AS I SHARED, WE DETERMINED TARGETED DRUGS AND THEN DID THAT SMALL DOUBLE BLIND PLACEBO CONTROL TRIAL. WHAT'S ANOTHER THING WE SAW REALLY INCREASE? HPV. WE'RE GOING TO LOOK AT ORAL HPV AND PEOPLE LIVING WITH HIV, AND IN CANCER. HPV IS HIGHLY SUCCESSFUL IN THE ORAL CAVITY. EVEN THOUGH THERE'S A VACCINE RIGHT NOW, THAT VACCINE IS TARGETING ADOLESCENTS, WHICH IS GREAT, AND IT'S A PREVENTIVE, SO THAT MEANS THAT 30 YEARS FROM NOW OR 40 YEARS FROM NOW WHEN THEY GET TO THE AGE WHERE CANCERS COME UP, THEY ARE NOT GOING TO GET THEM. BUT RIGHT NOW, WE STILL HAVE A BIG PROBLEM BECAUSE WE DON'T HAVE THERAPEUTICS. BACK IN 2008, WE SHOWED THAT HPV WAS DETECTED IN MORE THAN 80% OF NONSMOKERS AN NON-DRINKERS, ALL THESE PEOPLE COMING IN WHO HAD CANCER WHO JUST DIDN'T HAVE ANY RISK FACTORS. AND LOOKED FOR HPV THERE. AMONG THAT COHORT OF PEOPLE WERE TWO COUPLES, AND WHAT WE SAW, THE TWO COUPLES, ONE CAME IN WITHIN A YEAR OF EACH OTHER, THE OTHER ONE CAME IN WITHIN MAYBE SIX MONTHS OF THE OTHER. AND WHAT WE COULD SEE IN THOSE INDIVIDUALS WHO WERE COUPLES, THE FIRST COUPLE HAD BEEN TOGETHER FOR TEN YEARS. AND THE OTHER ONE HAD BEEN TOGETHER FOR ABOUT 15 YEARS. AND WE LOOKED AT THEIR TUMORS, COUPLE 1 AND COUPLE 2, THEY HAD THE SAME EXACT CHANGES ACROSS THE HPV GENOME WHICH SAYS THAT IT'S LIKELY THAT THIS VIRUS, THIS ORAL VIRUS THAT CAUSED THOSE CANCERS, WAS LIKELY TRANSMITTED ORALLY. THERE'S A POTENTIAL FOR ORAL HPV TRANSMISSION ASSOCIATED WITH ORAL CANCER DEVELOPMENT. WHEN WE LOOK AT HIV POPULATION, WE DID A CLINICAL TRIAL OF 400 HIV PATIENTS AND LOOKED PRE AND POST INITIATION OF HAART AND WHAT WE COULD SEE AS PEOPLE -- I HAVE A BETTER ONE HERE. AS PEOPLE'S CD4 COUNTS WENT UP WE SAW MORE HPV OVER TIME. SO WE HAD THESE PEOPLE WITH A MEAN CD4 COUNT INCREASE OF 125. THESE PEOPLE WERE PEOPLE WHO HAD MORE HPV, STATISTICALLY SIGNIFICANT. AND THEM TELLS US THAT EFFECTIVE IMMUNE CONTROL OF HPV IS NOT RECONSTITUTED BY ANTIRETROVIRAL THERAPY AND AT LEAST NOT DURING THE FIRST 24 WEEKS OF THERAPY. ONE THING THAT I MEANT TO SHARE WITH YOU, LET ME JUST SHOW THIS TO YOU HERE, IS THAT THE OTHER THING THAT WE SEE -- OOPS. THE OTHER THING WE SEE IS HIGH RISK TYPES PERSIST. WHY IS THIS IMPORTANT? IT'S BECAUSE THOSE TYPES ARE THE ONES THAT CAUSE CANCER. SO, THOSE HIGH RISK TYPES PERSIST IN THE CONTEXT OF HIV. WHICH MEANS THAT THESE IMMUNE SUPPRESSED FOLKS ARE GOING TO BE MORE AT RISK FOR HEAD AND NECK CANCERS AS WELL. OKAY. THE OTHER THING WE LOOKED AT WAS DOES THE VACCINE WORK IN PEOPLE WHO ARE IMMUNE SUPPRESSED? AND THIS WAS TRIAL A-5298. WHAT WE DID WAS TO ADMINISTER THE VACCINE TO HIV-POSITIVE MEN, AND WOMEN, WHO WERE OVER AGE 27. AND WHAT WE COULD SEE WERE THE VACCINES WORKED. THEY GOT GOOD ANTIBODY RESPONSES, OUT TO 18 MONTHS OF FOLLOW-UP, BOTH IN PEOPLE HIV POSITIVE AND HIV NEGATIVE. WHICH WAS GREAT. WE LOOKED AT 148 HIV NEGATIVE AND 75 HIV POSITIVE AND THEY HAD RECEIVED THREE DOSES OF THE QUADRIVALENT VACCINE. NOW WE HAVE MONOVALENT BUT EARLY ON THE ONE WITH THE FOUR SUBTYPES. AND EVERY SINGLE INDIVIDUAL, SEROCONVERTED, REGARDLESS OF THEIR HIV STATUS. THAT WAS GOOD NEWS. ONE THING FOR US TO PAY ATTENTION TO WITH REGARD TO ORAL HEALTH IS THAT POST VACCINATION, ORAL HPV 16 ANTIBODIES AT MONTH 7 WERE DIFFERENT BETWEEN POSITIVE AND NEGATIVE MEN AND WERE ACTUALLY LOWER IN THE HIV POSITIVE. SO WHY IS THIS SOMETHING WE NEED TO PAY ATTENTION TO? BECAUSE HPV 16 IS THE MAIN DRIVER FOR HEAD AND NECK CANCERS. SO WE HAVE TO BE VERY CAREFUL AND VERY DILIGENT TO MAKE SURE WE'RE LOOKING FOR ORAL CANCERS IN THESE IMMUNE SUPPRESSED INDIVIDUALS. SO, I STARTED TO THINK, WHY ARE THESE VIRUSES SO SUCCESSFUL? WHAT ELSE IS HAPPENING? OKAY? THEY ARE NOT IN THERE BY THEMSELVES. EVEN THOUGH AS SCIENTISTS WE OFTEN STUDY ONE ORGANISM AT A TIME, RIGHT? WHEN I WAS A GRADUATE STUDENT, I'M AN EBV PERSON, BUT WE DO NOT -- WE LIVE WITHIN MANY, MANY, MANY OTHERS INHABIT US, VIRUSES, FUNGI, BACTERIA. WE KNOW THAT ALMOST HALF OF U.S. ADULTS HAVE PERIDONTITIS AND THERE'S A RELATIONSHIP BETWEEN THAT AND CANCER. MAYBE THERE'S SOME EFFECT THAT VIRUSES HAVE ON BACTERIAL METABOLISM. OR PERHAPS, YEAH, THAT VIRUSES CAN ACTUALLY INDUCE EPIGENETIC CHANGES ON THE BACTERIA THAT MIGHT MAKE THINGS WORSE. OR, YOU KNOW, THIS INFLAMMATORY RESPONSE BY HAVING ALL THOSE ORGANISMS TOGETHER MAY DRIVE THINGS. BUT WE THOUGHT IT COULD BE THE OTHER WAY AROUND. AND HERE'S WHAT I SHOULD HAVE SHOWN FIRST. AND THAT'S ACROSS MANY STUDY TYPES, THERE ARE RETROSPECTIVE, LINKAGE, META-ANALYSIS, AND BOTTOM LINE UP TO FIVE-FOLD INCREASE IN HAVING CANCER IF YOU HAVE PERIDONTAL DISEASE. WHAT IS HAPPENING HERE? I APOLOGIZE THIS IS CUT OFF. WE DECIDED TO DO AN EPIDEMIOLOGIC ASSESSMENT AT UNC WITH ANN SANDERS, WHAT WE KNOW IS WE LOOKED IN THIS COHORT OF 6,000 PEOPLE FROM NHANES. OF THOSE, 7.4% OF THEM CARRIED ORAL HPV. WHEN WE STRATIFIED BY PERIOSTATUS, PEOPLE WHO DON'T HAVE PERIODONTITIS, A CLEAR RELATIONSHIP EPIDEMIOLOGICAL, ADJUST FOR AGE, RACE, NUMBER OF SEXUAL PARTNERS, SO THIS RELATIONSHIP LOOKS REAL. HERE'S THE OTHER THING WE SAW IF WE GO BACK TO OUR OWN COHORT THERE'S A MARKER THAT'S A PATHOGENIC BACTERIAL SENSOR, THE PEOPLE WHO ARE HPV POSITIVE THEY HAD HIGHER LEVELS OF THIS BACTERIAL SENSOR, AND WE SEE THERE'S A DISTINCT MICROBIOME ASSOCIATED WITH BEING HPV POSITIVE VERSUS NEGATIVE. SO WHAT IS GOING ON? WE ASKED THE QUESTION, CAN ORAL BACTERIA MODULATE THESE PROCESSES? WHAT HAPPENS WITH HPV IS THAT THERE'S OFTEN A MICRO ABRASION THAT OCCURS THAT ALLOWS THE VIRUS TO GET DOWN TO THE BASAL EPITHELIUM AND WHILE IN THIS STATE VIRAL EPISOMES CAN REPLICATE AND LOW RISK HPVs REPLICATE AND THEIR REPLICATION IS ASSOCIATED WITH EPITHELIAL DEVELOPMENT, OKAY? WE SEE ORAL WARTS DEVELOP. THEY ARE ASSOCIATED WITH LOW RISK HPVs. FOR HIGH RISK HPVs THEY CAN INTEGRATE VIRAL DNA OVEREXPRESSION AND CANCER-CAUSING DISEASE, WE ASKED THE QUESTION CAN ORAL BACTERIA MODULATE THESE PROCESSES? REPLICATION, ONCOGENE EXPRESSION, INTEGRATION? AND BASICALLY WHAT WE DID WAS TAKE SOME GOOD GUYS, LIKE KNOWN COMMENSALS AND BAD GUYS AT HIGH LEVELS, WE GREW THESE AND ISOLATED JUST THE END PRODUCTS, ADDED TO TUMOR VIRUS INFECTED DEVELOPS. HERE THE CELLS EITHER HAD EPISOMAL HPV IN THEM OR INTEGRATED HPV. ORAL BACTERIA, HERE LOOKING AT FN, HERE IT'S HPV 16 EPISOMES TREATED WITH MEDIA OR AS A CONTROL, WE CAN SEE, YES, CHECK THE BOX, FUSO BACTERIUM CAN PROMOTE. DANIELLE ASKED HOW IT CAN IMPACT IN THE CONTEXT OF CANCER. HERE LOOKING AT A CELL LINE THAT HAS AN INTEGRATED HPV IN IT ALREADY WHERE THE HPV LONG CONTROL REGION IS UPSTREAM OF THE E6 AND E7 ONCOPROTEINS. WHAT WE FOUND WAS WHEN DANIELLE TREATED THESE CELLS WITH P GINGIVALIS MEDIA WE SAW GLOBAL K9 ACETYLATION INCREASE. YOU CAN SEE INCREASED EXPRESSION OF GLOBAL K9, WE ADD INHIBITOR FOR P38 IT GOES AWAY. GOES DOWN. SO LOOKS LIKE THESE ARE P38 DEPENDENT. THE OTHER THING SHE DID WAS CHIP ASSAYS TO LOOK AT OCCUPANCY OF THESE EPIGENETIC MODIFICATIONS K9, K27, ACETYLATED HISTONE 3. PG DOES A GOOD JOB PROVIDING OCCUPANCY, WHEN YOU TREAT WITH AN INHIBITOR FOR P38 IT DROPS RIGHT DOWN. WE THINK THAT PG ACTIVATES P38 PATHWAY AND DOWNSTREAM OF THAT ARE ACTIVATING EPIGENETIC MODIFICATIONS. AND THEN WHAT HAPPENS AFTER THAT IS THOSE ACTIVATING EPIGENETIC MODIFICATIONS ALLOW FOR TRANSCRIPTION TO OCCUR, FOR THE BAD BY BUT NOT THE GOOD GUY. HERE IS PG, HERE IS STREP SANGUINEOUS, THREE-FOLD INCREASE IN E67 TRANSCRIPTS WITH GINGIVALIS, THIS LOOKS MORE LIKE THE MEDIA ALONE. WHAT WE DID, WHAT DANIELLE DID, WAS TO TREAT WITH THE P38 INHIBITOR AND LOOK DOWNSTREAM AT E6, E7 TRANSCRIPTS AND WE SEE WITH INHIBITOR A DECREASE, IT'S CLEAR P38 SIGNALING CAN BE MODULATED, P GINGIVALIS CAN INCREASE E67 KNOWN TO BE IMPORTANT DRIVER FOR ORAL CANCER SO THE ANSWER IS YES, ORAL BACTERIA CAN MODULATE HPV GENE EXPRESSION. NOW, I DON'T KNOW WHAT HAPPENED WITH ALL THESE WORDS BUT I'M GOING TO TELL YOU THAT JANET DOOLITTLE IN OUR GROUP WAS A BIOINFORMATICS PERSON AND SHE LOOKED ACROSS ABOUT 600 HPV CANCERS, IN THE PAPER LOOKED AT INTEGRATION SITES FROM, LET'S SEE, ABOUT 40,000 INTEGRATION SITES FOR HIV, ALSO LOOKED AT HBV, HCV, VIRUSES THAT INTEGRATE AND HPV BUT I'M SHARING HPV DATA NOW. WHAT SHE FOUND WERE THAT REPEATS IN OPEN CHROMATIN WERE CRITICAL. WE DIDN'T KNOW WHAT THINGS MODULATE INTEGRATION. BUT REPEATS ARE CRITICAL. OPEN CHROMATIN IS CRITICAL. IT SEEMS LIKE THIS IS NOT OCCURRING IN A RANDOM WAY. BUT THE OTHER THING YOU SEE IS THAT THERE ARE SPECIFIC EPIGENETIC CHANGES THAT OCCUR AT HEAD AND NECK SITES, WE'RE COMPARING HEAD AND NECK TO CERVICAL, CERVICAL IS BLUE, HEAD AND NECK INTEGRATION SITES ARE RED. EPIGENETIC CHANGES ARE REALLY IMPORTANT. WHAT I'M NOT SHOWING IS THAT WE COULD SHOW THAT THE SAME CHANGES OF THE BACTERIA PROMOTED WERE CHANGES THAT ARE IMPORTANT FOR INTEGRATION TO OCCUR. SO THERE'S THE OPPORTUNITY AND CHANCE THAT ORAL BACTERIA CAN MODULATE A DIFFERENT PART OF THE HPV LIFE CYCLE, REPLICATION, ONCOGENE EXPRESSION, HPV INTEGRATION ALL IMPORTANT TO CANCER. SO, THAT'S HPV. WE'LL TALK ABOUT -- ENDING WITH EBV, A MAJOR VIRAL PERIDONTAL PATHOGEN, MORE IN THE EAST THAN THE WEST. I'M CALLING IT A PERIDONTAL PATHOGEN, WE THINK OF A BACTERIA-DRIVEN DISEASE. AS YOU CAN SEE IN THIS -- THIS WAS AN ANALYSIS LOOKING ACROSS MULTIPLE STUDIES AND MULTIPLE COUNTRIES, JAMAICA, TURKEY, TAIWAN, JAPAN, WHERE HE LOOKED AT DIFFERENT LEVELS OF PERIODONTAL STATUS AND CONSISTENTLY YOU DETECT THESE VIRUSES. THIS FAMILY OF VIRUSES. HERPES SIMPLEX 1, EPSTEIN-BARR, WE SAW CMV WHEN WE HAD EBV SO WE'RE GOING TO FOCUS ON EBV. THIS IS A HUMAN GAMMAHERPES VIRUS, BIGGER THAN HPV, IT'S FOREVER, AND IMPORTANTLY IT'S ASSOCIATED WITH MULTIPLE HEAD AND NECK CANCERS INCLUDING BERKET'S LYMPHOMA, ORAL CAVITY, HODGKIN'S LYMPHOMA, AND EPITHELIAL MALIGNANCIES LIKE NASOPHARYNGEAL CARCINOMA, AND NOT IN THE HEAD AND NECK ARE GASTRIC CANCERS AND 10% OF GASTRIC CANCERS ARE CAUSED BY EPSTEIN BARR VIRUS. SO WE KNOW THAT THIS VIRUS WHILE IT'S SUPPOSED TO BE DORMANT AND USUALLY IS DORMANT IN OUR B CELLS, IT CAN BE REACTIVATED, REACTIVATION INITIATED BY EARLY PROTEINS, Z AND R. YOU CAN HAVE REACTIVATION OR LYTIC GENE EXPRESSION IN RARE B CELLS BUT IT OCCURS PRETTY SIGNIFICANTLY IN OROPHARYNGEAL EPITHELIAL CELLS, IMPORTANT BECAUSE THE HIGHER VIRAL LOAD FOR EBV, IT'S BEEN ASSOCIATED WITH HIGHER VIRAL LOAD SET POINTS DURING PERSISTENT INFECTION, ASSOCIATED WITH INCREASED MALIGNANCY RISK. NOW, JUST AS WE GO BACK TO THE FIRST GROUP OF PEOPLE, I'M GOING TO SHOW YOU THAT WHEN WE TREAT PERIDONTAL DISEASE IT MAKES A DIFFERENCE FOR THE VIRUS. THIS IS ONE PERSON'S MOUTH, A DISEASED SITE IN TAN, RED IS HEALTHY SITE. SO THIS PERSON HAD PERIODONTAL DISEASE, THIS WAS A HEALTHY SITE, LOOK HOW MUCH HIGHER THE COPY NUMBER IS THERE. WITH TREATMENT INTERVENTION, IT GOES DOWN. YOU ALREADY SAW THIS BEFORE. OUR TREATMENT WAS EFFECTIVE, PERIDONTAL IMPROVED LOOKING AT POCKET DEPTH AND BLEEDING ON PROBING OVER TIME BUT LOOK AT EBV, DROPS BY UP TO TWO LOGS. THE OVER PIECE OF EVIDENCE I LIKE TO SHARE, THERE'S A DISEASE THAT EBV IS ASSOCIATED WITH CALLED HAIRY LEUKOPLAKIA, THE ONLY PERMISSIVE -- ONLY PATHOLOGIC MANIFESTATION OF PERMISSIVE EBV INFECTION. WHAT YOU CAN SEE WHEN PEOPLE ARE IN CARE, WE SAW LESS HAIRY LEUKOPLAKIA THAN PEOPLE OUT OF CARE. WITH TREATMENT, WE'RE TREATING FOR PERIO, WE SEE THE INCIDENCE OR PREVALENCE OF HAIRY LEUKOPLAKIA DROP DOWN. THIS EBV-DRIVEN DISEASE GOES AWAY AS THE PERIODONTAL DISEASE IMPROVES. AND WHEN WE LOOK AT WHAT THE MICROBIOME LOOKS LIKE, IN PEOPLE WHO HAVE LOW EBV VERSUS HIGH EBV, WE CAN SEE THAT IT'S DISTINCT WHERE YOU'VE GOT THE BAD GUYS RIGHT UP TOP, AND PEOPLE HAVE HIGH EBV LOADS, OKAY, AND STREPTOCOCCUS IN THE LOW. YOU GOT TWICE AS MUCH BACTEROIDES, TWICE AS OF FUSO IN HEME WITH HIGH EBV COMPARED TO LOW, AND HERE THE HIGH EBV, NONE IN THE LOW. SO WHAT DO WE THINK IS HAPPENING HERE? NOW, I PROBABLY SHOULD HAVE DESCRIBED THIS EARLIER BUT WE KNOW THERE ARE PIONEER BACTERIA, LIKE THE GOOD GUY, STREP SANGUINEOUS WE'RE USING HERE, EARLY COLONIZERS, THESE ALLOW ACQUISITION OF BRIDGING BACTERIA AND THE BIOFILM CAN ACCUMULATE KEYSTONE PATHOGENS LIKE P GINGIVALIS. THERE'S GOT TO BE A HOST RESPONSE. SO CELLS ARE GOING TO COME IN, MACROPHAGES, LYMPHOCYTES COME IN TO FIGHT BACTERIAL INFECTIONS. SOME OF THESE GUYS MAY BE INFECTED THEMSELVES, CMV CAN BE CARRIED IN MACROPHAGES, LYMPHOID CELLS CAN CARRY SARCOMA VIRUS, WHAT HAPPENS WHEN CELLS WHO COME IN TO TAKE CARE OF THE PERIO DISEASE AND HAPPENING TO CARRYING A VIRUS, WHAT HAPPENS? SO OUR HYPOTHESIS HERE IS THAT THESE CELLS WHEN THEY MIGRATE INTO THE AREA WHERE THERE'S A SIGNIFICANT POLYMICROBIAL BACTERIAL INFECTION, THEY COME THERE AND THEY HAVE LATENT VIRUS, OKAY? WHEN THEY GET INTO CONTACT WITH THE PRODUCTS OF THE BAD GUYS, WE HIGH HYPOTHESIZE THAT ALLOWS FOR ACTIVATION AND ENHANCED ONCOGENIC PHENOTYPE. BASICALLY LIKE WE DESCRIBED FOR HPV WE DID THE SAME THING. BUT WHAT I CAN SHOW YOU IS WHEN WE PROVIDE THESE END PRODUCTS HERE, TO EBV-INFECTED CELLS, WE CAN SEE THAT THE GOOD GUY DID NOT ALLOW FOR EXPRESSION OF THE LYTIC PROTEIN, R, BUT THE BAD GUYS FN AND PG BOTH IN COME BIN DID A GOOD JOB ALLOWING FOR LYTIC ACTIVATION AND AS DETERMINED BY DETECTION OF R. SO, WHAT IS HAPPENING? YOU KNOW, IS WHAT WE'RE THINKING TRUE? THE FIRST THING THAT WE DID WAS WE TREATED LYMPHOID CELLS, B CELLS, OKAY, WITH THE SPENT MEDIA TO SEE WHAT WOULD HAPPEN. AND WHAT WE CAN SEE HERE IS ALL BAD GUYS ARE NOT CREATED EQUAL. HERE'S PREVOTELLA, IT'S NOT DOING ANYTHING. PG DOES A PRETTY GOOD JOB OF ALLOWING THIS EARLY PROTEIN Z TO BE MADE AND FN DOES AS WELL. BUT THE GOOD GUY DOESN'T. SO TO TEST THE HYPOTHESES TREATED LYMPHOID CELLS, GOT REACTIVATION. THESE LYMPHOID CELLS HAVE A GFP, EBV WITH GFP IN THE GENOME. SO YOU CAN ACTUALLY SEE THEM HERE. AND WE CO-CULTURED THEM WITH NORMAL ORAL KERATINOCYTES AND LEFT THEM THERE FOR 24 HOURS. WASHED THEM TO REMOVE ANYTHING THAT WAS UNATTACHED. AND WE ACTUALLY SEE THAT WE HAVE PROMOTION OF ADHERENCE OF THESE B CELLS WHEN THE BUG IS THERE, WHEN FN WAS THERE, OKAY? THEN WE SEE THE VIRUS THAT THESE ORAL PATHOGENS ARE ACTUALLY UPREGULATING, FUSION PROTEINS FROM THE VIRUS, GH AND GL, THESE TRANSCRIPTS ARE SIGNIFICANTLY INCREASED WITH PG AND FN, AND NOT SO MUCH WITH PREVOTELLA INTERMEDIA OR FS. AFTER 24 HOURS OF CO-CULTURE WE CAN SEE THAT THE VIRUS INITIALLY WAS IN THE B CELLS, AND THEN WITH THE PRESENCE OF THAT BACTERIA THOSE B CELLS WERE STUCK. WE LET IT CO-CULTURE FOR 72 HOURS AND START TO SEE THE VIRUS MOVE FROM B CELLS TO THE EPITHELIAL CELLS. WE HAVE HYGRAMICIN, WE SEE TRANSFER TO THE EPITHELIAL CELLS. SO, WE KNEW THAT THIS IS A PATHOGEN THAT REPLICATED IN EPITHELIAL CELLS, WE KNEW THAT IT WAS LATENT IN B CELLS. WE DID NOT KNOW THAT BACTERIA COULD ACTUALLY HELP MOVE THE INFECTION FROM THE B CELL INTO THE EPITHELIAL CELLS. WELL, WHAT HAPPENS ONCE IT GETS THERE? WE SEE BACTERIAL PATHOGENS CAN ACTUALLY DRIVE A NUMBER OF PATHWAYS, SO HERE WE LOOK AT THE PHOSPHO-PROTEOME, WE CAN SEE THERE'S A VERY DISTINCT PHOSPHO-PROTEOME COMPARED TO THOSE ASSOCIATED WITH MEDIA ALONE. WITH RNAseq WE SEE THE SAME STORY AS WE SEE WITH PHOSPHO-CHI N OME ANALYSIS. CYTOKINE SIGNALING, NF-kappaB. I CALL THIS THE FIREWORKS PLOT FROM THE PHOSPHO-KINOME ANALYSIS, SIGNAL TRANSDUCTION AND MODULATION OF CELL CYCLE ARE SIGNIFICANTLY MODULATED. WE EXPECT THE CELL CYCLE MIGHT BE MODULATED, THAT'S GOOD FOR THE VIRUS, THE VIRUS GETS GOING, THE VIRUS IN ORDER TO REPLICATE NEEDS TO TAKE ADVANTAGE OF THE CELL CYCLE. WE FOCUS ON SIGNAL TRANSDUCTION, LOOKS LIKE GPCR SIGNALING IS INITIATED BY PG, AND THIS PREDICTED BACTERIA MEDIATED ACTIVATION OF THE NF-kappaB PATHWAY AND ERK LIKE WITH PHOSPHO-KYNOME. WE DID ADDITIONAL WORK I WON'T SHOW, TO PROVE PATHWAYS WERE BEING USED. WHAT WAS INTERESTING WAS WHEN WE TREATED WITH INHIBITORS WE SAW THAT EPIGENETIC MARKS WERE DIMINISHED AS WELL. SO WE THEN ASKED, WELL, DO THESE PATHOGENS ACTUALLY MODIFY CHROMATIN? WE GOT TOGETHER WITH NATE HATHAWAY OVER AT THE CANCER CENTER AT UNC AND HE HAD A CHROMATIN ASSAY WHERE HE CAN ADD RAPAMYCIN AND PUSH A SYSTEM TO HETEROCHROMATIN, SO WHEN YOU DON'T HAVE RAPAMYCIN YOU'RE IN THE ACTIVE STATE, WE CAN SEE PEAKS IN ACTIVE STATE AND COMPARE DIFFERENT SITUATIONS HERE, EITHER TREATING WITH COMMENSAL OR PATHOGENS, SO WHEN WE ADD RAPAMYCIN WHAT WE SEE HERE IS THAT WE MOVE WITH PG AND FN INTO AN ACTIVE STATE, FROM THE HETEROCHROMATIN. SO THE CLOSED CHROMATIN STATE REMAINS WITH COMMENSAL IN GREEN MORE MEDIA ALONE BUT FN AND PG MOVIE TO OPEN CHROMATIN STATE. WE CAN SEE BASED ON INHIBITION FN, PG AND COMBINATION DO A PRETTY GOOD JOB THERE. AND SO HERE I'M SHOWING YOU A COUPLE, WE'RE LOOKING AT K 27 ACETYLATION, K 4 TRIMETHYLATION, A CHIP ASSAY, LOOKING AT FOLD INCREASE IN K 27 ACETYLATION OCCUPANCY. AND WHAT WE SEE HERE, ON THE PROMOTER OF Z OR R FOR EBV, NO CHANGES WITH MEDIA ALONE OR COMMENSAL, A LITTLE WITH FN BUT LOOK AT PG. PG SIGNIFICANTLY INCREASING OCCUPANCY OF K 27, IT'S NOT JUST THE VIRUS. THERE ARE A BUNCH OF CELLULAR GENES WHERE WE CAN SEE INCREASED INCREASED OCCUPANCY OF K 27. PREVOTELLA DOESN'T DO SUCH A GOOD JOB, S S, NO AT ALL, BUT FN AND PG DO A GOOD JOB. IN ORDER FOR K4 TO GET TO TRIMETHYL STATE FROM DIMETHYL STATE, THE MLL COMPLEX REQUIRES ASH 2L TO MOVE TO TRIMETHYLATION. WE MADE shRNA, TARGETED ASH 2L, AND WHAT YOU CAN SEE IS WHEN WE TREAT WITH THE MEDIA ALONE NOTHING IS HAPPENING, WHETHER SCRAMMED BELLED OR SR RNA, TREAT WITH PG WE CAN SEE GOOD H3K4 TRIMETHYLATION WITH THE SCRAMMED BELL BUT NOT shRNA. AND WHAT'S REALLY INTERESTING OR COOL TO ME IS NOT ONLY IS THAT shRNA PULLING DOWN OR DIMINISHES THE K4 TRIMETHYL, Z GOES DOWN AS WELL. CRITICAL AND IMPORTANT TO THESE IMMEDIATE EARLY GENES. WE LOOK AS CROSS EBV TRANSCRIPTOME IN LYMPHOID AND EPITHELIAL CELLS, AND WE TREAT WITH EITHER PATHOGENS OR COMMENSAL. ORANGE MEANS HIGH LEVELS OF GENE EXPRESSION, PURPLE IS DOWNREGULATED. IN LYMPHOID CELLS PG AND FN DO A GOOD JOB AT UPREGULATION ACROSS THE GENOME. IT'S MORE CONSERVATIVE IN THE EPITHELIAL CELLS. OKAY. THOSE GENES DOWNREGULATED INTERESTINGLY ARE GENES IMPORTANT TO INFLAMMATION. BUT WHAT'S REALLY INTERESTING TO ME IS THE COMMENSAL IS DOWN-MODULATING THESE THINGS. WHAT DOES THIS MEAN? A HEALTHY MICROBIOME THAT IS ATTAINED BY GOOD ORAL HYGIENE CAN SHIFT WHAT'S HAPPENING NOT ONLY WITH THE BACTERIA BUT WITH THE VIRUSES. AND SO NOT SO WE SAW -- WE SAW CHANGES AT THE LEVEL OF THE GENOME IN TERMS OF TRANSCRIPTOME, WE CAN ALSO SEE THIS INCREASE IN COPY NUMBER THAT'S PRETTY SIGNIFICANT BUT NOT WITH THE COMMENSAL. BUT ALSO MAJOR PROCESSES IMPORTANT TO CANCER. WE SEE INCREASE IN HIF-1 ALPHA, BCL-2, ANTI-APOPTOTIC GENE, WHEN WE LOOK ACROSS EMT, MESENCHYMAL, SHOOT, PATHWAYS, WE ACTUALLY SEE THEY ARE HIGHLY UP, OKAY, WITH PG. OKAY. EPITHELIAL MESENCHYMAL TRANSITION, OKAY? AND WE CAN SEE THAT BACTERIA-DRIVEN EBV CAN LEAD TO INCREASED INFLAMMATION. THE WORSE PERIODONTAL DISEASE, THE MORE LIKELY THE VIRUS WILL BE THERE. THESE ARE LYMPHOID CELLS WITH EBV IN THEM, AND WE'VE GOT PARENTAL CELLS WITH NO EBV AND THEN CELLS WITH EBV. ONES WITHOUT EBV IN ORANGE, WITH EBV IN BLUE. WE TREAT AND SO WHEN WE TREAT CELLS THAT DON'T HAVE EBV IN THEM, WITH LTA, THREE OR FOUR INCREASE IN AL BUT BUT WITH EBV UP TO 60-FIELD, INTERFERON BETA TWO OR THREE FOLD, GOES UP TO 120-FOLD INCREASE. THAT VIRUS MAKES A DIFFERENCE. WHEN WE LOOK ACROSS DISEASE AND FUNCTION WE SEE CHANGES BUT OTHER PROCESSES VERY IMPORTANT TO CANCER, ORGANIZATIONS OF CYTOPLASM, MICRO TUBULES, INVASION, MEDIATED BY ORAL BACTERIA. KEY FINDINGS FOR EBV THESE BAD GUY BACTERIA CAN PROMOTE ACTIVATING EPIGENETIC MARKS, ACTIVATE POWERFUL PROLIFERATIVE SIGNALING PATHWAYS CENTRAL TO ONCOGENESIS AND NOT ONLY INCREASE IN VIRUS BUT CANCER-ASSOCIATED CELLULAR GENES PROMOTING EBV-ASSOCIATED MALIGNANCY, THE GOOD GUY NOTHING HAPPENS, IT GOES ON ABOUT ITS BUSINESS, YOU DON'T HAVE THESE PROBLEMS. SO JUST TO CONCLUDE, FOCUSING ON THE DIFFERENCE THAT ORAL CARE MAKES, CARE IS ASSOCIATED WITH A LOT OF THINGS THAT MAKE A DIFFERENCE TO A PATIENT. LOWER ORAL AND SYSTEMIC INFLAMMATION, CONTEXT OF HIV, HIGHER CD4 COUNTS, LOWER VIRAL LOADS, LOWER OPPORTUNISTIC INFECTION, MORE COMMENSAL BASED MICROBIOME. HAVING A PATHOGENIC MICROBIOME NOT ONLY DRIVES HPV BUT CAN ALSO DRIVE EBV BY ACTIVATING A NUMBER OF DIFFERENT CELLULAR PATHWAYS, ACTIVATING DOWNSTREAM EPIGENETIC MARKS, AND BY PROMOTING THE TRANSCRIPTION AND VIRAL REPLICATION THAT MOVES THINGS FORWARD TOWARDS DISEASE. BUT WE KNOW THE COMMENSALS CAN ACTUALLY DOWNREGULATE AND SUPPRESS THESE PROCESSES. SO THE BOTTOM LINE, ORAL HEALTH CAN MAKE A DIFFERENCE. OKAY. SO I JUST WANT TO ACKNOWLEDGE MY GROUP. THIS IS MY GROUP. AND OUR GROUP IS CALLED VOICE, VIRAL ORAL INFECTIONS AND IMMUNE SUPPRESSION IN CANCER, AND THESE ARE THE PEOPLE IN MY GROUP, COLLABORATORS, AND OUR SUPPORT FROM NIDCR AND HRSA. AND I'D LIKE TO THANK ALL OF YOU AND HAPPY TO TAKE ANY QUESTIONS. [APPLAUSE] >> REALLY GREAT TALK. ONE QUESTION ABOUT THE DIFFERENT BACTERIA. HAVE YOU THOUGHT OF LOOKING AT METABOLOMICS, WHAT IS INSIDE MEDIA? >> WE'VE STARTED THOSE STUDIES AND DONE MASS SPEC, A BUNCH OF FRACTIONATION, IT LOOKS LIKE THERE ARE IMPORTANT COMBINATIONS BUT IT'S NOT COMING DOWN TO A SPECIFIC THING. WE TESTED A BUNCH OF BIOPHYSICAL AND BIOCHEMICAL PROPERTIES, WE KNOW IT'S HEAT TOLERANT, WE KNOW IT'S LESS THAN 3,000 KILODALTONS, AND THE MASS SPEC TOLD US ABOUT SOME OF THE THINGS THAT ARE POTENTIAL -- THE POTENTIAL MEDIATORS BUT WE'RE WORKING ON THAT. >> THANK YOU. >> YEP. >> THANK YOU FOR SHARING THAT BEAUTIFUL COMPELLING STORY ABOUT THE POWER OF ORAL HEALTH AND HYGIENE ON PHYSICAL HEALTH. I HAVE A QUESTION FROM VIDEOCAST FROM MATT HOFFMAN. GREAT TALK. DO YOU KNOW HOW PG INFLUENCES HPV EPIGENETIC TRANSCRIPTION? IS IT SOMETHING THE PG SECRETES OR THE CARBOHYDRATE CODE? >> IT'S SOMETHING THAT'S SECRETED INTO THE MEDIA. SOMETHING THAT'S SECRETED AND CERTAINLY DOES ALLOW FOR ACTIVATING EPIGENETIC MARKS THROUGH P38 PATHWAY. >> VERY GOOD. A FOLLOW-UP, I'M DYING TO KNOW AFTER YOU TOLD THIS BEAUTIFUL STORY ABOUT EBV, IF YOU THINK IT'S A PHYSICAL INTERACTION WITH THE PG AND FUSO BACTERIA MAKING CHROMATIN MARKS OR IN THE PATHWAY OF SECRETED PRODUCT. >> WHEN WE TREAT WITH INHIBITORS TO SIGNALING PATHWAYS, IT GOES AWAY. SO IT LOOKS LIKE IT'S AN ORPHAN GPCR RECEPTOR, THAT'S BEING ENGAGED, FOR EBV NF-kappaB AND ERK 1 WITH PATHWAYS ENGAGED, TREAT WITH INHIBITORS, CANONICAL OR NON-CANONICAL NF-kappaB OR ERK 1, ACTIVATING MARKS DON'T OCCUR ANYMORE. >> COMPELLING EVERYDAYENS. THANK YOU. >> ANOTHER QUESTION FROM THE VIDEOCAST. FANTASTIC PRESENTATION. I HOPE THIS PRESENTATION IS ON VIDEOCAST AND CAN BE MADE AVAILABLE TO THE PUBLIC. YES, IT WILL BE, SHOULD BE POSTED WITHIN THE NEXT COUPLE DAYS, AVAILABLE TO THE PUBLIC. >> THANK YOU. THANK YOU ALL FOR COMING. [APPLAUSE]