MY NAME IS DIMA HAMMOUD. SENIOR INVESTIGATOR. TODAY WE HAVE A PROGRAM THAT INCLUDES A DISTINGUISHED GROUP OF SCIENTISTS AND PHYSICIANS FROM ALL OVER THE WORLD WHO HAVE AGREED TO SHARE WITH US THEIR CUTTING EDGE RESEARCH. AS YOU MIGHT KNOW INFECTIOUS DISEASE IMAGING IS A FAST GROWING FIELD. BUT THE INTEREST IS NOT NEW -- WE HAVE STRIVED AS A MEDICAL COMMUNITY TO PROVIDE FAST AND ACCURATE DIAGNOSIS OF INFECTIOUS DISEASES WHICH WAS BOLSTERED BY THE PROGRESS WE HAD WITH IMAGING WITHOUT THESE INCLUDING CTM/MRI. MOLECULAR APPLICATIONS STARTED TO ATTRACT MORE ATTENTION AND IF YOU ACTUALLY -- CHECK WE HAVE HAD A GREAT DEAL OF DISCOVERIES AND INCONVENIENCES. RELATED TO IMAGING AS YOU'LL SEE IN THE FIRST SESSION AS WELL AS TARGETING THE PATHOGENESIS SPECIFICALLY LIKE YOU'RE GOING TO SEE IN THE SECOND AND THIRD SESSION TODAY AND BETTER UNDERSTANDING THE PATHOGENESIS LIKE YOU'LL SEE IN OUR LAST SESSION. SO WHY IS IMAGING -- SO IMPORTANT. THE FRESHER MEMORY IS THAT OF THE COVID-19 PANDEMIC. YOU CAN EASILY NOTICED THE ACCELERATING RATE OF PANDEMICS AND EPIDEMICS OVER THE LAST 100 YEARS OR SO. ALL OF THIS CULMINATED IN THE DEVASTATING EVENTS WHERE A TINY VIRUS OFFENDED OUR LIVES AND KILLED MORE THAN 3.5 MILLION PEOPLE WORLDWIDE. SO THERE ARE NO GUARANTEES THAT NEW VIRUSES OR OTHER SUPER PATHOGENESIS WILL NOT BE AFFECTING US IN THE FUTURE AND WE ALL NEED TO BE READY FOR OUR MEXICO INDIVIDUAL. -- OUR NEXT COVID. MORE THAN 8 MILLION PEOPLE DIE AND ALMOST 400,000 YEARS OF LIFE ARE LOST DUE TO INFECTIONS ACROSS THE GLOBE. INFECTIONS ARE THIRD IN MORTALITY AND FIRST IN MORBIDITY AMONG ALL HUMAN DISEASES. IT MIGHT SURPRISE YOU TO KNOW THAT -- SEPSIS IS RESPONSIBLE FOR KILLING ONE OUT OF-FOUR PEOPLE GLOBALLY. A HUGE GLOBAL CONCERN IS ANTI ANTIMICRO RESISTANCE. AND HOW ABOUT CANCER PATIENTS? A LOT OF YOU DO CANCER RESEARCH. INFECTION IS THE SECOND LEADING CAUSE OF DEATH. AND THE IRONY WITH ALL OF THE NEW THERAPIES REVOLUTIONIZING CANCER THERAPIES -- THE RATES OF DEADLY INFECTIONS ARE INCREASING SIGNIFICANTLY AND SPEAKING OF FUNGAL INFECTIONS WHICH IS MY FAVORITE TOPIC IT TURNS OUT THOSE AFFECT OVER A BILLION PEOPLE GLOBALLY WITH ALMOST 1.5 MILLION DEATHS ANNUALLY. THOSE ARE JUST A FEW FACTS OF HOW INFECTIOUS AFFECTS US IN THE WORLD. AND BECAUSE OF ALL I JUST MENTIONED IT'S ABSOLUTELY IMPERATIVE THAT WE ADVANCE THE FIELD OF MOLECULAR IMAGING. PERHAPS THE GREATEST VALUE WE GET IS THE ABILITY TO GET MORE SPECIFIC AND FASTER DIAGNOSIS OF VARIOUS PATHOGENESIS. WE WOULD BE ABLE TO QUANTIFY DISEASE -- FOR EXAMPLE AND MAYBE DIAGNOSE INFECTIONS WITH OUT THE NEED FOR INVASIVE PROCEDURES WHICH ARE QUITE EXPENSIVE AS WELL. IT WOULD BE GREAT TO ASSESS TO DETERMINE THE OPTIMAL DURATION OF THE TREATMENT AND TRY TO HELP OVERCOME THE RESISTANCE AND IT WOULD BE REALLY MORE HELPFUL FOR US IN FIGHTING THE INFECTIONS NONINVASIVELY USING IMAGING. BUT WE CANNOT DO THIS ALONE. WHAT IS NEEDED IS A MULTI-DISCIPLINARY APPROACH. AND MOST IMPORTANTLY WE NEED YOUNG SCIENTISTS TO MOVE THE FIELD FORWARD. SO WITH CONFERENCES LIKE THE ONE TODAY WHAT WE'RE TRYING TO DO IS EXPOSE YOU TO EXAMPLES OF CUTTING EDGE RESEARCH TARGETING INFECTIONS AND INFLAMMATION AND HOPE THIS WILL INCREASE YOUR INTEREST IN THE FIELD AND HELP YOU UNDERSTAND THE EXCITING POTENTIAL AND JOIN US IN THIS CRUSADE. SO NOW TO THE PRACTICAL DETAILS. THIS IS A WHOLE DAY SYMPOSIUM. I HOPE YOU WILL STICK AROUND. THE SESSIONS ARE VERY, VERY INTERESTING. AND FEEL FREE TO TAKE A SNAPSHOT OF THE SCREEN IF YOU DON'T HAVE THE AGENDA HANDY. THE SECOND AND THIRD FOCUSES ON TRACKING THE PATHOGEN WHILE THE LAST IS IMAGING VIRAL INFECTIONS AND HIGH CONTAINMENT PATHOGENESIS AND SOME HOUSEKEEPING NOTES. IF YOU WOULD LIKE MORE INFORMATION ABOUT OUR SPEAKERS AND TODAY'S AGENDA YOU CAN USE THE LINK LISTED ON THE SLIDE. ALSO THROUGHOUT THE TALKS YOU CAN SUBMIT QUESTIONS USING THE OPTION ON THE LEFT SIDE OF THE VIDEO CAST SCREEN. WE'LL COLLECT YOUR QUESTIONS AND ANSWER THEM DURING THE LAST 10 MINUTES OF EACH SESSION AND IF WE DO NOT GET TO YOUR QUESTIONS FOR TIME LIMITATIONS WE'LL MAKE EVERY EFFORT TO PROVIDE YOU AN ANSWER BY E-MAIL. THE VIDEO CAST WILL BE CAPTIONED. AND WITHOUT ANY FURTHER DELAY I WOULD LIKE TO START OUR PROGRAM -- AND WELCOME OUR FIRST SPEAKER OF THE DAY DR. MA DR. MATTHIAS MIEDERER. >> SO DR HAMMOUD THANK YOU VERY MUCH FOR INVITING ME TO THIS TALK. AND THE PANDEMIC SITUATION HAS TAUGHT US HOW TO DEAL WITH VIDEO CONFERENCES AND THIS IS BECOMING EFFICIENTLY AND YOUR SYMPOSIUM IS STARTING IN THE MORNING AND I'M HERE IN GERMANY IN THE MIDDLE OF THE WORKING DAY. AND I WOULD LIKE TO GIVE AN OVERVIEW OF OUR WORK. I'M A NUCLEAR MEDICINE PHYSICIAN WORKING IN CLINICAL ROUTINE BUT WE'VE GOT -- AN ESTABLISHED PRECLINICAL LABORATORY WHICH SHIFTED RECENTLY -- IT'S FOCUSED ON IMAGING THE IMMUNE SYSTEM TRIGGERED BY THE ADVANCE IN IMMUNE ONCOLOGY WHERE IF THE IMMUNE SYSTEM IS TAILORED AND CHANGED TO FIGHT CANCER AND IN THIS FIELD THERE HAS BEEN AN INCREASED INTERESTING -- IN TECHNOLOGY TO IMAGE THE IMMUNE SYSTEM AND I SEE THERE IS A LOT OF OVERLAP WITH THE TOPIC OF YOUR SYMPOSIUM. WHAT WE SEE IN CLINICAL PRACTICE TYPICALLY AS NUCLEAR MEDICINE PHYSICIANS IS IMAGING OF THE GLUCOSE METABOLISM. YOU'LL SEE ON THE LEFT-HAND SIDE PATIENT WITH MALIGNANT -- WHICH SHOWS THE TYPICAL ENLARGED GLUCOSE METABOLISM WHICH IS THE BASES OF A LOT OF ONCOLOGY DIAGNOSTIC HOWEVER IT CAN BE INCREASED IN INFLAMMATION. YOU CAN SEE A SIMILAR PICTURE WITH A PATIENT THAT SUFFERS FROM ACTIVE TUBERCULOSIS WHERE SIMILAR MECHANISMS ARE TRUE WHERE THE GLUCOSE METABOLISM IS HIGHLY ENLARGED WHICH IS A FIRST START. AND RECENTLY DUE TO THE COVID-19 VACCINATION WE SEE IN MANY PATIENTS THESE ACTIVATED LYMPH NODES WHICH ARE DUE TO VACCINATION AND MUST NOT BE CONFUSED WITH MALIGNANT LYMPH NODES AND THIS RNA TECHNOLOGY IS EXTREMELY POTENT TECHNOLOGY THAT CAN BE USED FOR A VARIETY OF APPLICATIONS. AT OUR CENTER OF COURSE WITH PFIZER -- THE RNA TECHNOLOGY IS ALSO USED IN VACCINATION AGAINST CANCER AND THESE EFFECTS CAN BE SEEN ON ROUTINE PET. IF YOU SEE A PATIENT THAT HAS BEEN INTRAVENOUSLY INJECTED WITH mRNA VACCINE THE SPLEEN SHOWS AN ENHANCED INFLAMMATION. WHAT WE'VE LEARNED FROM THE PATHWAYS OF THE OF THE INFLAMMATION THAT THIS IS -- TRIGGERED. IT'S NOT THE T-CELL. HOWEVER THIS WIDELY AVAILABLE GLUCOSE -- HAS PROMPTED US TO WORK WITH IT AND TO INCUBATE IMMUNE CELLS WITH -- AND YOU CAN SEE THEY ARE NICELY TAKING UP FDG AND THEY ARE LABELED WITH THIS RADIOACTIVITY AND AFTER INJECTION INTO A MOUSE AS YOU SEE HERE ON THE LEFT HAND SLIDE THESE CELLS CAN BE IMAGED AND THE MIGRATION AT LEAST IN THE FIRST FEW HOURS CAN BE IMAGED BY SMALL ANIMAL PET. HOWEVER THE HALF LIFE OF THIS 18 IS TOO SHORT TO IMAGE THE LONG PROCESSES THAT ARE SEEN IN THE IMMUNE SYSTEM AND IT'S NOT OF GREAT VALUE TO IMAGE ONLY THE FIRST FEW HOURS. THEREFORE WE NEED LONGER LIFT RADIO ISO TOPES. AND THIS CAN BE LABELED TO THE ANTIBODY AND HALF LIFE OF THE RADIO ISOTOPES AND THE ANTIBODIES ARE FITTING WELL TOGETHER. WE USED THE ANTIBODY AS AN IMAGING A LITTLE THAT WAS LABELED AND IT CAN BE INCORPORATED AND YOU'VE GOT A TRACER THAT CAN BE SPECIFICALLY IMAGE CELLS AND IN THIS CASE WE AIM TO IMAGE THE T-CELLS TO GET AN OVERVIEW OF THE T-CELL POOL AND MIGRATION OF T CELLS AS SUCH AND WHAT ARE USED AS A MODEL SYSTEM WAS -- HUMANIZED MOUSE MODEL WHERE WE TRANSPLANTED HUMAN IMMUNE CELLS AND THIS MICE THAT NOT ONLY LACK T AND B CELLS BUT THEY ALSO LACK CERTAIN -- -- SITE KIND CASCADES. -- CYTOKINE CASCADES. IT IS SIMILAR TO THE GRAPH -- WHAT SEE IN LEUKEMIA PATIENTS AND SERVES AS A GOOD MODEL TO IMAGE THE HUMAN IMMUNE SYSTEM IN THIS ENVIRONMENT. THE METHODS AND TECHNICAL SET UP FOR THIS EXPERIMENT IS THAT HUMAN -- CELLS ARE PURIFIED AND THEN INJECTED INTO THE IMMUNE COMPROMISED MICE AND AFTER A COUPLE OF DAYS OF MIGRATION AND PROLIFERATION OF THIS IMMUNE CELLS THE RADIOACTIVE LABEL ANTIBODY IS INJECTED AND AFTER A CERTAIN TIME OF ONE TO SEVEN DAYS THE IMAGING CAN OBTAIN THE SITUATION OF THE HUMAN IMMUNE SYSTEM IN THIS MOUSE. IN A FIRST PILOT STUDY WE USED THE AFORE-MENTIONED TECHNOLOGY WHERE WE PRELABELED THE HUMAN IMMUNE CELLS FDG BY INCUBATING THEM AND WASHING THEM AND INJECTING THEM INTO THE CAVITY AND AS YOU CAN SEE HERE THE HUMAN IMMUNE CELLS THEY DISTRIBUTE VERY RAPIDLY IN THE CAVITY AND AT THE END OF THE IMAGING WHICH IS DUE TO THE LIMITED HALF LIFE OF THE F18 YOU CAN SEE THAT THESE IMMUNE CELLS GET A LITTLE BIT ATTACHED TO THE INTESTINE AND ALSO TO THE LIVER. AND WE CORRELATED THIS MODEL WITH THE IMMUNE CHEMISTRY AFTER THE INJECTION. YOU CAN SEEP THE CELLS HERE IN THE FORM OF CD45 IMMUNO CHEMISTRY NOT ATTACHED IN THE CAVITY ADJACENT TO FAT TISSUE BUT WITHIN A FEW HOURS YOU CAN SEE ALSO THE IN FILTRATION EITHER IN THE INTESTINE OR A LITTLE BIT LATER ALREADY INTO THE LIVER TISSUE WHICH IS THEN THE BASES FOR THE LATER DEVELOPING -- DISEASE. WHAT WE ALSO SEE IN IMMUNO CHEMICAL HISTORY THERE ARE A LOT OF T CELLS PR -- PROLIFERATED WHICH ARE SOMETIMES WHY THE T CELLS ATTACK THE LEVEL OF THE MOUSE AND CAUSE THIS DISEASE. HOWEVER, WE HAVE GOT A LOT OF IMMUNE CELLS. WE'VE GOT ALL LINEAGES OF THE IMMUNE CELLS. WE SEQUELS THAT HAVE THE CELL MARKERS BUT ALSO T HELPER CELLS AND SITE OH TOXIC CELLS. WE SEE MICRO FACIAS AND NATURAL KILLER CELLS. THAT REFLECTS THAT WE'VE GOT HERE A MODEL SYSTEM THAT REALLY REFLECTS THE WHOLE HUMAN IMMUNE SYSTEM IN THIS MODEL AND THEN WHEN WE IMAGE THE ANTIBODY WE SEE A SIMILAR IMAGE IN THE FIRST FEW HOURS OR IN THE FIRST DAYS WHERE YOU SEE THIS SPOTS OF IMMUNE CELLS MAINLY LIMITED TO PEAR TOE KNEEL CAVITY. TERRIFIC TOE KNEEL CAVITY. PERFECT TOE KNEEL CAVITY. -- AND YOU CAN SEE ALSO THAT THERE IS BACKGROUND THAT WE'VE GOT SOME ACCUMULATION IN THE LIVER AND HEART DUE TO A CIRCULATING ANTIBODIES. AND IF YOU COMPARE NOW THE IMAGING WITH THE TC IMAGING TO THE CD3 IMAGING WITH THE FDG IMAGING YOU CAN SEE THE CONTROL MOUSE WITH THE DISTRIBUTION BACKGROUND IN LIVER AND UNSPECIFIC ENRICHMENT IN THE SPLEEN YOU DON'T SEE ANY SPECIFIC UP TAKE ON FDG IN THE NORMAL MICE IS TYPICALLY SEEN MOSTLY IN THE HEART. IF YOU'VE GOT AN ANIMAL THAT HAS GOT A HUMAN TRANSPLANTED IMMUNE SYSTEM YOU SEE THIS HIGHLY ENLARGED -- ELEVATED UPTICK IN THE LIVER AND THIS MULTIPLE INFLAMMATION SPOTS BOTH IN LIVER AND IN THE PERITONEAL CAVITY IN. CONTRAST YOU SEE THE FDG IS A LITTLE BIT -- CHANGE IN COMPARISON TO CONTROL BUT THIS IS MORE OR LESS AN UNSPECIFIC EFFECT DUE TO THE DEVELOPING HOST DISEASE. AND THIS IS AGAIN ONE EXAMPLE WHERE WE SEE THIS INFLAMMATORY SPOTS AND HOW THIS CORRELATE WITH IMMUNO HISTORY YOU SEE THE BACKGROUND IMAGING FROM THE DISTRIBUTION AND YOU SEE NO CD3 TARGETS ON IMMUNO CHEMISTRY IMAGERY AND ON THE OTHER SIDE YOU SEE THESE FEW MILLIMETER SPOTS OF CLUSTERED T CELLS THAT REFLECT EXACTLY WHAT WE SEE IN THE SMALL ANIMAL. NOW WHAT WE DID NEXT AFTER WE WERE FEELING THAT THIS CAN BE NICELY IMAGED TO APPLY THIS METHOD TO IMAGE EFFECT THE IMMUNE SYSTEM AND IN THE FIRST APPLICATION WE TRIED TO SUPPRESS THE EFFECT OF IMMUNE REACTION AND TO SUPPRESS THE GRAPH HOST DISEASE BY PURIFYING REGULATORY CELLS FROM THE DONOR AND INJECTING THE T-CELLS WITHIN IMPLANTATION AND AS YOU CAN SEE ON THE DEVELOPMENT OF THE ANIMAL WEIGHTS THAT AFTER INJECTING WITH THE T REGULATORY T-CELLS TOGETHER WITH THE HUMAN IMMUNE SYSTEM THE ANIMALS GET MUCH BETTER AND THE WEIGHT CHANGES ARE NOT THAT SEVERE IN THE UNCHANGED GROUP. IT IS NOT TOTALLY SUPPRESSED SO WE'VE GOT SOME RESAID CALL DISEASE COMPARED TO THE NORMAL DISEASE AND YOU CAN SEE ALSO WHEN YOU LOOK AT THE MAIN TARGET -- THE LIVER ENZYME ARE STILL ELEVATED IN MICE THAT HAS BEEN TRANSLATED WITH THE HUMAN IMMUNE CELLS SYSTEM TOGETHER WITH PURIFIED REGULATORY T-CELLS. WE HAVE THIS CONTROL PET AGAIN WITH A VERY SLOW BACKGROUND AND YOU SEE THIS HIGHLY UP TAKE WITH THE PET GRAPH WITH THE HOST MANIFESTATION WHICH NICELY CORRELATES ALSO AGAIN WITH THE IMAGING WHERE YOU SEE MAINLY THIS INFLAMMATORY SPOTS IN VARIOUS AREAS IN THE LIVER AND IF YOU COMPARE IT THEN IN THE GROUP THAT HAS BEEN SUPPRESSED -- WITH THE REGULATORY T-CELLS YOU SEE MUCH FEWER SPOTS AND THE BACKGROUND OF THE T-CELLS ARE NICELY SUPPRESSED AND THIS CAN ALSO BE SEEN IN A MUCH LOWER UP TAKE IN THIS PET SCAN AND WE CAN SHOW THAT THIS APPLICATION OF THIS IMMUNE IMAGING CAN IMAGE SOMETHING WHICH IS MORE OR LESS ONLY SEEN ON HISTOPATH ALLERGY AND THE QUANTIFICATION OF THE CD3 CELLS TRANSLATE NICELY TO THE IMAGING WE SEE WITH THE SMALL ANIMAL PET. I CONCLUDE THAT WE'VE GOT METHOD THAT CAN BE EASILY APPLIED IN SMALL ANIMAL AND ALSO POTENTIAL FOR CLINICAL TRANSLATION AND I WILL THANK YOU IN ADVANCE FOR ALL DISCUSSION AND ALL QUESTIONS AND I WOULD LIKE ALSO TO ACKNOWLEDGE THE MAIN PEOPLE WHO DID THIS WORK IN OUR LABORATORY AND THEN I THANK YOU AND GIVE BACK TO DR. HAMMOUD FOR THE MODERATION. >> THANK YOU VERY MUCH. THIS WAS A WONDERFUL PRESENTATION. BEFORE I PROCEED I WANT TO REMIND EVERYBODY THAT YOU CAN SUBMIT QUESTIONS USING THE SEND LIVE FEEDBACK OPTION ON THE VIDEO CAST. SO OUR SECOND TALK TODAY WILL BE GIVEN BY DR. PATRICK HUGHES WHO'S AN ADJUNCT SENIOR FELLOW IN AUSTRALIA. >> OKAY. THANK YOU. FOR THE INVITATION TO GIVE THIS PRESENTATION AND YES, IT IS A LITTLE BIT LATE HERE BUT I HAVE HAD MY COFFEE SO WE'LL GET THROUGH. JUST BY WAY OF INTRODUCTION I LEFT MY POSITION AND I'M NOW AT ADELAIDE BIOLABS. SO MY INTEREST IN IMAGING STARTED FROM INFLAMMATORY BOWEL DISEASE AND MY BACKGROUND IS MORE RELATED TO LOOKING AT THE IMMUNOLOGY OF BOWEL DISEASE AND THEN I MOVED TO A NEW BUILDING AND STARTED TO GET MORE INVOLVED WITH THE TEAM THAT IS MAKING RADIO LABELS AND ALSO IMAGING. SO I'LL TAKE YOU THROUGH SOME OF THE BACKGROUND OF INFLAMMATORY BOWEL DISEASE AND MOVING INTO THE WORK THAT WE'VE DONE HERE. SO INFLAMMATORY BOWEL DISEASE IS CHARACTERIZED BY SYMPTOMS OF BLOODY DIARRHEA, PAIN AND WEIGHT LOSS AND OCCUR IN THE PRESENCE OF READILY AVAILABLE PATHOLOGY. THERE ARE TWO MAJOR TYPES. THERE IS ULCER COLITIS. THEY DON'T TYPICALLY GO INTO THE SMALL INTESTINE. THEY ARE MOVING IN A FLAT SHAPE AND THEY ARE TYPICALLY RESTRICT SO THEY ARE QUITE SHALLOW. IN CONTRAST WITH CROHN'S DISEASE IT CAN OCCUR ANYWHERE WITHIN THE GASTROINTESTINAL TRACT. BUT THEY ARE NOT CONTIGUOUS. PERIODS OF HEALTHY TISSUE BETWEEN THE LESIONS. ANTHE DIFFERENCES INDICATE THAT THE TYPE OF IMMUNE RESPONSE AND THEREFORE THE CAUSES LIKELY DIFFER. HOWEVER WE DON'T KNOW WHAT THESE CAUSES ARE. THE CURRENT THINKING IS THAT THEY OCCUR ON THE BACK OF A COMPLEX GENETIC PREDISPOSITION AND THIS LAYS TO A LOSS OF IMMUNE TOLERANCE AND UNKNOWN ENVIRONMENTAL FACTORS THAT PROVIDES A HYPERSENSITIVE IMMUNE RESPONSE TO NORMALLY TOLERATED ANTIGEN. IT'S FAIRLY PREVALENT. THIS IS AN OLD STUDY NOW BUT IT WAS OBSERVED TO OCCUR IN ABOUT ONE IN EVERY 250 PEOPLE. ABOUT THE SAME AS SCHIZOPHRENIA AND TYPE ONE DIABETES AND THE REVERENCE IPREVALENCE IS INCREASING WORLDWIDE. TREATMENT CAN WORK AND LEAD TO REMISSION BUT WE HAVE PROBLEMS WITH TREATMENTS. THERE IS A LACK OF EFFICACY. AND ALSO A TOLERANCE WHERE WE GET SECONDARY NONRESPONDERS OR NONRESPONSE AND THIS OCCURS WHEN ANTIBODIES ARE GENERATED AGAINST DRUGS. THIS LEADS TO RELAPSE AND REPEATED BOUTS OF REMISSION AND THEN LEADS TO FIBROSIS. SURGERY OCCURS IN ABOUT 15%. BUT IT'S NOT CURATIVE FOR CROHN'S. THE LESIONS MAY REAPPEAR. SO HOW ABOUT THE DIAGNOSIS OF IBD. IT'S HEAVILY RELIANT ON ENDOSCOPY WHICH IS INVASIVE. A CAMERA THAT IS PUSHED UP THROUGH TH WRECK TOMORROW OR DOWN THROUGH THE MOUTH. IT DOES NOT PROVIDE ANY REALTIME ASSESSMENT. IT'S A VISUAL AID. YOU'RE ONLY SEEING WHAT THE CAMERA CAN SEE. SO THAT LED US TO THINK BEFORE USING ANTIBODIES AS TOOLS AND immunoPET WHICH I'M SURE YOU'RE FAMILIAR WITH. IT RELIES ON THE DETECTION OF RADIO LABELED FDG. WE'RE LOOKING AT ACTIVITY. NOT SPECIFIC FOR MEDIATORS WHEREAS IMMUNO PETROLEUM CAN PROVIDE FOR SPECIFICITY. OUR FIRST STEADY WE WANTED TO SEE IF WE COULD IMAGE INFLAMMATION IN A MOUSE. WE HAVE A MODEL. IT'S BEEN AROUND FOR DECADES. AND BASICALLY 2% -- DSS IS ADDED TO DRINKING WATER AND THE MICE DRINK IT. AFTER FIVE DAYS WE SWITCHED THE DSS OUT FOR WATER AND YOU CAN SEE THE BODY WEIGHT CONTINUES TO DROP. SHORTENING OF THE COLON LENGTH IS A SIGN OF INFLAMMATION AND YOU CAN SEE HERE THE COLON IS NOT SHORTER IN THE DSS TREATED MICE AND WE'VE GOT SOME CHAMBERS WHERE WE CAN MEASURE THE PHYSIOLOGICAL -- INDICATING THAT WE HAVE INFLAMMATION. WE'VE GOT A MODEL. SO THE NEXT THING WE DECIDED TO LOOK AT IS WHETHER IT WAS BETTER TO IMAGE MEDIATORS OR IMMUNE CELLS. WE LOOKED AT IMMUNE MEDIATORS BECAUSE THERE IS A FEATURE OF ACTIVE INFLAMMATION. WE HAVE THE IMMUNE MEDIATORS MPO AND ALSO ONE BETA. ONE BETA IS KEY. BUT MORE IMPORTANTLY FOR A STUDY LIKE THIS IT'S REDUCIBLE. SO THERE IS NOT MUCH BATE PA FLOATING AROUND IN A NORMAL HEALTHY PERSON BUT IT GETS SWITCHED ON WHEN INFLAMMATION IS ACTIVE. WE ALSO LOOKED AT IMMUNE CELLS. WE'RE LABELING IT POSITIVE. AGAIN SORRY IT LABELS -- NEWT FILL. THE FIRST ONE IS THE IMMUNE MEDIATOR. AND INJECTED INTO THESE MICE. WE CAN SEE AS THE VIDEO SCANS THROUGH THE COLON SHOWS UP QUITE LIGHT AND WHEN WE COMPARE THAT AGAINST THE DSS MICE AS WE TRACK THROUGH WE CAN SEE A SIGNATURE -- AND WHEN WE DID VOLUME OF ANALYSIS VOLUME OF INTEREST ANALYSIS YOU CAN SEE THERE IS QUITE A LARGE INCREASE IN THE DSS TREATED MICE. AND WHAT WE ALSO TRIED TO DO IS CORRELATE THIS IMAGE WITH SYSTEM SEVERITY. THIS IS THE SICK MICE AND THE VOLUME OF INTEREST WITH A PERCENTAGE OF BODY WEIGHT LOSS AND WE GOT A FAIRLY STRONG TREND TOWARDS THE CORRELATION. SO NEXT WE LABELED UP THE BAY AND DID THE SAME THING. PUT THEM INTO THE DSS TREATED MICE AND HEALTHY CONTROLS AND WE CAN SEE THE SIGNATURE INDICATING THAT WE CAN SEE THE CD11B POSITIVE CELLS BUT NO CORRELATION. THERE IS NOTHING CLOSE THERE. AND THEN WHEN WE LOOKED AT VOLUME OF INTEREST STUDIES LOOKING AT ALL OF THE ORGANS IN THE MUSIC SEE THE BETA LABELED WITH ZIRCONIUM IS RESTRICT TO THE INTESTINAL TRACK. THERE IS NOTHING IN THE OTHER TISSUES WHEREAS THE CD11B THAT LABELS THE INNATE IMMUNE CELLS WAS ALSO PRESENT IN THE INTESTINE, COLON AND SMALL INTESTINE BUT REALLY WENT EVERYWHERE THROUGHOUT THE BODY. THAT TOLD US THAT THE ZIRCONIUM IS MUCH MORE RESTRICTED THAN THAT LABELED TO CD11B AND THIS IS IMPORTANT BECAUSE WE WANT THE -- GOING TO THE SOURCE OF INFLAMMATION AND NOT THROUGHOUT THE BODY. TO SUMMARIZE THAT STUDY IMMUNO PET -- DIRECTED AGAINST KEY INMATE IMMUNE MARKERS DETECTS INFLAMMATION IN THE COLON. AND THIS MAKES ACCEPTS. IT IS CONCENTRATED AT THE SITE. SO WHAT ABOUT FIBROSIS. IT'S THE MOST COMMON COMPLICATION. IT'S A CONSEQUENCE OF REPEATED INFLAMMATION AND HEALING. YOU HAVE A WHOLE CASCADE OF EVENTS GOING ON HERE. A HOST OF DIFFERENT IMMUNE CELLS CELLS. AND A HOST OF OTHER MEDIATORS ARE INVOLVED THERE AND I THINK IT'S WORTH NOTING THAT THIS IS BASICALLY A GENERAL OVERVIEW OF FIBROSIS IN ANY TISSUE OF THE BODY. NONE OF THESE PATHWAYS SEEM TO BE SPECIFIC ABOUT THE GUT AT ALL. AND I THINK WHAT THAT IS TELLING US IS THAT WE DON'T KNOW ENOUGH ABOUT FIBROSIS TO DISTINGUISH THESE PATHWAYS BETWEEN DIFFERENT TISSUES. BUT MOVING ON -- THERE IS NO TREATMENTS FOR FIBROSIS. SURGE REAP THE ONLY OPTION AND THERE IS POOR DETECTION METHODS ESPECIALLY FOR THE GUT AND ESPECIALLY FOR THE EARLY SIGNS OF FIBROSIS WHICH START BENEATH THE LAYER THAT THE ENDOSCOPE CANNOT REACH. SO THERE IS A ROLE FOR immunoPET HERE. SO THE FIRST THING IS ESTABLISH A MODEL. WE'VE GOT THE SIMILAR MODEL OF FIBROSIS. FOLLOWED BY THREE DAYS OF WATER. THESE ARE INFLAMED MICE. AND THEN IN OTHER GROUP OF MICE WE EXTEND THE HEALING PERIOD TREATED WITH WATER OUT FOR ANOTHER WEEK AND THEN TREAT AGAIN WITH DSS AGAIN A LONGER PERIOD OF WATER FOR THREE CYCLES. TRYING TO REMITTING AND RELAXING. AND AT THE END OF THIS -- THIS IS BASICALLY -- -- FOR COLLAGEN AND YOU CAN SEE THE COLON LENGTH RECOVERS BUT IT IS STILL MUCH SMALLER THAN THE HEALTHY MICE. HYDROXY PRO LEAN OR COLLAGEN SIGNIFICANTLY INCREASED. >> SO WE LOOKED AT CYTO KINDS. -- PEOPLE THAT ARE SUSPECTED OF HAVING INFLAMMATORY BOWEL DISEASE PASS A STOOL SAMPLE WHERE IT'S MEASURED. A FECAL SIGN OF CURRENT INFLAMMATION. SO WHAT YOU CAN SEE -- WITH ALL OF THESE CYTOKINES IS THAT THEY ARE ALL INCREASED IN INFLAMED MICE EXCEPT FOR R10 WHICH IS SUPPRESSIVE. THERE IS NO DIFFERENCE BETWEEN THESE MARKERS. THE SAME WITH THE FECAL MPA AND THEY ARE NOT SUITABLE FOR TARGETS FOR FIBROSIS WHICH FURTHER ESTABLISHED OUR MODEL BECAUSE INFLAMMATION SEEMS TO HAVE SUBSIDED. THE NEXT THING IS THE MMPs. YOU CAN SEE EACH OF THE MMPs THAT WE LOOKED AT -- THERE WAS NO ANTIBODY AVAILABLE FOR MOUSE MMP9. THAT IS A NEGATIVE REGULATOR. AND AS YOU CAN SEE THEY ARE ALL INCREASED IN INFLAMED MICE. WHEN WE LOOK AT THE FIBROIDIC MICE -- THAT REMAINED NOT SIGNIFICANTLY DIFFERENT TO THE INFLAMED MICE. SO WE LOOKED AT USING PRO MMP9 AS AN IMAGING TOOL IN PET. SO THE FIRST THING WE NEEDED TO DO IS MAKE SOME FAB FRAGMENTS BECAUSE IF YOU WANT TO GO INTO THE CLINIC THEY ARE VERY LARGE. THEY ARE GREAT DRUGS. BUT AS IMAGING AGENTS THEY HAVE LONG CLEARANCE TIME SO YOU'RE INCREASING THE RADIATION THAT THE BODY MAY SEE. ALSO THE FC REGION BINDS NONSPECIFICALLY SO YOU WANT TO GET RID OF THAT AND WE CAN DO THAT -- AND WHAT YOU'RE LEFT WITH THE FAB ANTIBODY FRAGMENTS SO THESE FRAGMENTS ARE THE -- WITH THE BINDING SPECIFICITY. THEY ARE MUCH SMALLER BECAUSE WE GOT RID OF THAT FC PORTION AND NONSPECIFIC BINDING AND THE CLEARANCE TIMES ARE REDUCED AND YOU DON'T HAVE RADIO LABELED -- ANTIBODY. SO WE MADE THE FAB FRAGMENTS AND PURIFIED THEM AND CONGREGATED THEM. AND STUCK THEM IN THE MICE. HERE WE HAVE THE CONTROL MICE AND THE FIBROIDIC MICE AND WE SEE AN INCREASE IN THE COLON IN THE FIBROIDIC MICE BUT WE ALSO SAW IT IN THE KIDNEY AND THAT WAS CAUGHT UNEXPECTED. SO WE VALIDATED THE COLON DATA WITH -- SPECTRA IMAGING AND ALSO LOOKED AT BIODISTRIBUTION AND WHAT YOU CAN SEE HERE IN THE FIBROIDIC MICE WE HAVE THE INCREASE IN THE COLON. ALSO INCREASES IN THE SPLEEN AND THE LIVER BUT IT'S THE KIDNEY THAT WAS HUGELY THE SIGNAL WAS HUGE. SO THIS WAS COMPLETELY UNEXPECTED. THERE IS NOTHING REALLY ABOUT KIDNEY DAMAGE IN THIS MODEL IN THE LITERATURE SO WE WENT BACK AND HAD A LOOK TO SEE WHETHER THE KIDNEY BECOMES FIBROIDIC IN THIS MODEL AS WELL AGAIN USING THE HSS. THE CONTENT IS INCREASED IN THE FOR BRIDEIC MICE AS IS IF THE PRO MMP -- PROTEIN. SO WE DON'T REALLY UNDERSTAND THE MECHANISMS. SOB FAIR WITH -- PATIENTS DON'T GENERALLY COMPLAIN TO KIDNEY SYMPTOMS AND IT MIGHT BE RELATED TO THE MODEL WHERE IT'S IN THE DRINKING WATER THEREFORE GOING THROUGH THE KIDNEY BUT WHAT I THINK IT REALLY SHOWS IS THAT THE BENEFIT AND IMAGING THE WHOLE BODY THAT PET PROVIDES. SO JUST TO CONCLUDE WE'VE SHOWN THAT IN TH -- IN THE FIBROSIS MODEL -- MMPs ARE INCREASED BUT DECREASED WITH RESPECT TO INFLAMMATION. WHEN WE GENERATED THE FAB FROGMEANTS WE SHOWED THAT THEY CAN DETECT FIBROSIS AND GENERALLY RESTRICT TO THE COLON BUT ALSO DETECTS KIDNEY FIBROSIS. WHAT THIS SHOWS IS THE -- TO DETERMINE THE INFLAMMATORY SITES TO DIAGNOSE PATIENTS WITH SYMPTOMS TO DETECT FIBROSIS AND ALSO TO TRANSLATE THIS ACROSS TO OTHER DISEASES BECAUSE AT THE MOMENT THE MECHANISMS OF FEW BRINGSES SEEM TO BE SHARED AND THAT LEAVES ME TO THANK MY LAB -- AND ALSO COLLABORATORS AND FUNDING SOURCES AS WELL. THANK YOU. >> THANK YOU VERY MUCH DR. HUGHES. THIS WAS REALLY EXCITING WORK. VERY INTERESTING AND THANK YOU AGAIN FOR DOING THIS FROM AUSTRALIA AT SUCH A WEIRD HOUR. >> YEAH. I'VE HAD MY COFFEE. >> THAT'S GOOD. OKAY IT'S MY PLEASURE TO INTRODUCE OUR THIRD SPEAKER OF THE DAY. DR. DORIANE McGavern. SENIOR INVESTIGATOR AND CHIEF -- AT NINDS, NIH. DORIANE. >> ALL RIGHT. I REALLY APPRECIATE THE OPPORTUNITY TO PRESENT HERE THIS MORNING ON ONE OF MY FAVORITE TOPICS WHICH IS IMAGING CNS INFECTIONS AND THE IMMUNOLOGY OF THOSE INFECTIONS SO TODAY I'M GOING TO HONE ON TWO BASIC MECHANISMS BY WHICH IMMUNE CELLS DESTROY OR DAMAGE CNA VASS VASCULATURE. THIS HAS SOME RELEVANCE TO WHAT IS PROBABLY HAPPENING IN COVID PATIENTS ALTHOUGH THAT IS NOT THE INFECTIOUS DISEASE THAT I WILL BE SHOWCASING. THE CNS COMPARTMENT IS NUANCED. THE VAS VASCULATURE VARIES. IT TENDS TO RESIDE IN THE LINING OF THE BRAIN AND I'LL SHOW THAT IN THE NEXT SLIDE HERE. THERE IS ALSO A BLOOD BRAIN BARRIER THAT TRIES TO KEEP THINGS OUT OF THE CNS AND THE -- IT IS SURVEYED BY MICRO GLEE A. -- SORT OF THE OUTER MOST LATER THE DAUGHTER WHAT MATTER IS PART OF THE ME THEN GEESE. * IT HAS OPEN BLOOD VESSELS ELSE AND DRAINAGE AND A LOT OF IMMUNE CELLS. THIS OPEN STRUCTURE IS DEFENDED BY A FULL ARMAMENT OF IMMUNE CELLS. ONCE YOU GET BELOW THE ARACANOID MATTER THIS IS A MEMBRANE HERE AND BENEATH THAT THERE IS A VESSEL. THIS IS LAYING ON THE PIA MATTER. IT IS CLOSED UP AND THE IMMUNOLOGY IS MUCH LESS COMPLEX ONCE YOU ENTER THE SPACE HERE AND THEN AS VESSELS DESCEND THEY GET WRAPPED BY PROCESSES AS WELL AND THIS BECOMES THE PROTO TIPIC BLOOD BRAIN BARRIER. THERE IS NOT A NOT OF IMMUNOLOGY DOWN HERE. THIS IS TYPICALLY IN ACTED BY MICRO GLEE A. * AND THERE IS A A LITTLE LESS SO HERE IN THE SUB A RACK NOTICED SPACE. *. TYPICALLY WHEN CELLS ENTER YOUR CELLS SOMETHING HAS GONE WRONG. YOU HAVE AN INJURY, A SERIOUS INFECTION. MAYBE DAMAGE TO THAT PARTICULAR COMPARTMENT THAT WARRANTS THESE CELLS COMING IN. THERE IS A LOT OF INTERESTING IMMUNE CELLS UP IN THE DURHAM MATTER BECAUSE THEY NEED TO SURVEY THIS COMPARTMENT. TODAY I'M GOING TO HONE -- [ INDISCERNIBLE ] [ AUDIO DIFFICULTIES ] THAT IS THE IMMUNE CELLS PHYSICALLY CAUSING THE DAMAGE TO THE BLOOD VESSELS AND I'LL SHOW YOU HOW WE ACCOMPLISH THAT IN THE COMING SLIDES. SO THIS IS A SCAN FROM A PHOTON MICROSCOPE OF THIN SKULL BONE AND I'LL SHOW YOU HOW WE DO THAT PREP IN THE NEXT SLIDE. THIS IS DOWN TO ABOUT 20 MICRONS AND UNDER THAT -- IS THE MOUSE AND THEN THE -- AND THEN UNDERNEATH THAT WE HAVE THE BRAIN -- THERE IS A LOT OF EXCITING STUFF THAT HAPPENS IN THIS SPACE. AND AGAIN SORT OF THE WORSE CASE SCENARIO IS YOU HAVE INFECTIOUS AGENTS THAT TRAVERSE THIS AND ENDS UP IN THE BRAIN -- AT WHICH POINT YOU HAVE TO WORRY ABOUT THE PATHOLOGY AND THE DAMAGE THAT THAT MIGHT CAUSE TO NEURONS. THIS SPACE IS CHOCK FULL OF LOTS OF STUFF. YOU'LL FIND STROMA -- YOU'LL FIND TONS OF MACRO FACIAS AND * AND ACTIVE IMMUNE CELLS IN THAT SPACE AS WE. HOW WE GET ACCESS USING TWO PHOTON. ONE APPROACH IS TO THIN THE BONE AND THIS LITTLE CIRCLE HERE IS ABOUT ONE TO ONE AND A HALF MILLIMETERS IN DIAMETER SO WE'RE TALKING ABOUT THE TIP OF A PENCIL. AND AGAIN ON TOP OF THAT WE ADD SOME ARTIFICIAL SPINAL FLUID AND WE DIP THE LENS INTO THIS SOLUTION AND VISUALIZE THROUGH THE BONE INTO THE MENENGIS AND INTO THE BRAIN -- THIS IS A STANDARD APPROACH AND WE'RE CAREFUL WHEN THINNING THE BONE HERE AS TO NOT CAUSE DAMAGE UNDERNEATH. A DISEASE AGAIN THAT I WANT TO SHOWCASE IS ONE THAT IS CAUSED BY IN THIS CASE A VIRUS. THIS CAN BE CAUSED BY BACTERIA THAT GETS INTO THE LINING OF THE BRAIN AND THEN TRIGGER A RESPONSE. SO THIS IS MENTA MENINGITIS. SO WE INFECT WITH THIS VIRUS. THE VIRUS THEN SORT OF COVERS THE LINING -- IT LOVES THE MENENGIS. *. THIS IS A BRAIN OF ASYMPTOMATIC ANIMAL ON DAY SIX AND THIS IS THE VIRUS. LINING THE MATTER OF THE BRAIN TISSUE. AND THEN THE IMMUNE ATTACK FOLLOWS. IT ENDS UP BEING THE IMMUNE RESPONSE THAT CAUSES THE DESTRUCTION. SO WE BECAME EXCITED ABOUT THESE CELLS BECAUSE WE THOUGHT HEY THEY ARE GOING TO BE THE ONE THAT'S DRIVE THE WHOLE PATHOLOGY PROBABLY BY KILLING STUFF. SITE OH TOXIC -- IS KNOWN FOR KILLING THINGS. THIS WAS A MOUSE THAT WAS ASYMPTOMATIC ON THE LEFT ON THE FIFTH DAY AND WE'RE IMAGING THE ME THEN GIVES. WE'VE LABELED THAT * WITH QUANTUM -- THAT WE'VE PUT INTO THE BLOOD SUPPLY AND THE T CELLS ARE TAGGED WITH GREEN FLUORESCENT PROTEIN. THEY ARE CAPABLE TO REACTING TO -- AND SO THEY ARE SWIMMING AROUND HERE IN THE SPACE AND LITERALLY ONE DAY LATER WHEN THE MICE GET SYMPTOMATIC AND THEY DIE FROM THE DISEASE WE SEE THIS. WE SEE ALL OF THESE CAT CELLS AND OPENING UP THESE MEN GEIAL VESSELS. SO, ITS A BRAINSTEM DISEASE CAUSED BY A BUILD UP IN PRESSURE. THIS CELLS WERE NOT CAUSING THE DISEASE BY CALLING VIRUS INFECTED TARGETS. THIS BRINGS ME TO THE MECHANISM WHICH I'LL SHOWCASE. IT IS THE RECRUITMENT OF OTHER CELLS THAT ENDS UP DRIVING THIS DISEASE. SO WE TERMED THIS -- BECAUSE YOU'RE LOOKING AT A VESSEL AND YOU SEE MONO CITES AND THEY COME AND EXTRACT FROM IT SIN NOWLY. -- SYNCHRONOUSLY. AND THAT VESSEL OPENS UP AND STARTS LEAKING CONTENTS AND WE SEE WHEN YOU HAVE OVERLY EXUBERANT -- VERSES BREAK OPEN AND START TO LEAK INTO THE SURROUNDING ME ANYONE GEESE. THIS IS A REALLY GENERALIZABLE CONCEPT OF HOW CELLS CAN DAMAGE A VESSEL AND IT IS BY EXSTRAFF GETTING BY OPENING THE DOORS THAT THEY ACCOMPLISH THIS. SO THAT IS ONE DESTRUCTIVE PROCESS BY WHICH I NATE IMMUNE CELLS CAN CAUSE DAMAGE. T-CELLS CAN CAUSE THE SAME TYPE OF DISEASE BUT WITHOUT THE HELP OF THE -- CELLS. THIS IS RELEVANT TO THE COVID-19 BRAIN PATHOLOGY THAT IS OBSERVED. THE DISEASE -- IS CEREBRAL MALARIA. PRIMARILY AFFECTS CHILDREN. I'LL THROW A FEW FACTS HERE ON THE RIGHT-HAND SIDE BUT BASICALLY IN CHILDREN THAT GET MALARIA YOU CAN HAVE A COMPLICATION. AND IT'S CALLED CEREBRAL MALARIA. IT'S ALREADY BAD ENOUGH THAT YOU GOT MALARIA FROM THIS PARASITE -- YOU GET IT FROM A MOSQUITO BITE AND THEN YOU'VE GOT THE PROBLEM OF RED BLOOD CELLS BEING INFECTED BY THIS PARASITE BUT AT THE SAME TIME YOU HAVE SOMETHING HAPPENING IN YOUR BRAIN. AND SO YOU CAN SEE IN THIS BRAIN HERE THERE ARE THESE LITTLE PUNTA. THESE ARE HIM AGES. AND THAT IS A FOCA FOCAL HEMORRHAGE. A 20% MORTALITY RATE. YOU'RE GOING TO HAVE -- AND IT IS CHARACTERIZED BY SEE QUEST RACIAL OF RED BLOOD CELLS ALONG THE VESSEL WALL. YOU CAN HAVE * VASCULAR BREACH AND SWELLING AND THE SWELLING IS THE CAUSE OF DEATH. AS THE BRAIN FILLS UP WITH FLUID THIS PUTS PRESSURE ON THE BRAINSTEM AND PATIENTS AND MICE WILL SUCCUMB TO THIS DISEASE SO I'LL SHOW YOU WHAT THAT LOOKS LIKE IN A HUMAN. THIS IS A PAPER PUBLISHED SEVERAL YEARS AGO SHOWING IN CHILDREN THE PRIMARY NODE IS BRAIN SWELLING. THIS IS A CHILD ON THE LEFT THAT DOESN'T HAVE THE SWELLING AND ON THE RIGHT HERE YOU CAN SEE THAT THERE IS ALL OF THIS DIFFUSE PATTERN NOW TO THE MRI IMAGE. THERE IS A DISTINCTION OR SWELLING OF THE TISSUE AND NOW PRESSURE BEING PLACED ON THE BRAINSTEM WHICH IS SHOWN HERE. SO COMPARE A TO C AND YOU CAN SEE THERE IS A NOTABLE DIFFERENCE IN THE AMOUNT OF FLUID IN THIS COMPARTMENT SO WHEN THE BRAINSTEM GETS PRESSED ON. THIS WILL AGAIN CAUSE A FATALITY AND THAT IS CALLED HERNIATION. THE MODE OF DEATH. SO IN TERMS OF A MOUSE IT'S SIMILAR. THE DISEASE INSTEAD OF IT BEING CAUSED BY -- IT'S CAUSED BY CLASS MODEIAN -- IN RODENTS AND YOU CAN INFECT MICE WITH RED BLOOD CELLS AS SHOWN HERE ON THE BOTTOM AND SIX DAYS LATER THEY ARE GOING TO START TO SHOW EVIDENCE OF HELP. RAGE. THERE IS PROFOUND PATHOLOGY AS THE TWO RED THINGS HERE. AND BASICALLY THE MICE WILL LOSE THEIR SENSE OF SMELL AND THEY WILL ALSO DIE FROM BRAINSTEM HERNIATION. SO THE AMOUNT OF LEAK IS DISPLAYED HERE WITH THE SMURF BRAIN HERE ON THE LOWER RIGHT AND WHAT WE'VE DONE IS INJECTED EVANS BLUE INTO THE BLOOD SUPPLY AND IN A NORMAL MOUSE IT'S A CLEAN BRAIN AND IN CONTRAST HERE ON THE RIGHT YOU CAN SEE THAT THE BLUE IS LEAKING EXTENSIVELY THROUGHOUT THE BRAIN TISSUE. SO A MOUSE HERE WILL GO FROM NO EVANS BLUE TO ALL OF THE EVANS BLUE AND WATER CONTENT IN THE MOUSE'S BRAIN WILL INCREASE FROM ABOUT 78% WHICH IS OKAY UP TO ABOUT 79% AND THAT IS FATAL. THAT IS TOO MUCH WATER IN THE BRAIN AND IT CANNOT BE ACCOMMODATED BECAUSE OF THE SKULL BUILD. SO WHAT DOES THAT LOOK LIKE PATHOLOGICALLY. WE CAN SEE THAT HERE IN A HISTOLOGY SESSION. AND IN THIS EXPERIMENT WHAT WE DID IS WE INJECTED EVANS BLUE AS A TRACER. AND WE ALSO INJECTED -- IODINE TO LABEL DEAD CELLS AND YOU CAN SEE HERE THAT THE OLFACTORY BULL' AND THE BRAINSTEM ARE LIT UP WITH EVANS BLUE AND PRO PID YUM IODINE. WE THOUGHT BASED ON WHAT WE HAD SEEN IN OUR MODEL OF VIRAL MENINGITIS THAT THE -- THAT THESE CELLS WERE GOING TO BREAK OPEN BLOOD VESSELS. MUCH THE SAME WAY THAT WE SAW IN OUR MODEL OF VIRAL MENINGITIS. AND THIS THEORY PROVED TO BE INCORRECT. IT'S AN ENTIRELY DIFFERENT MODE OF PATHOGENESIS AND DRIVEN BY CD8 T-CELLS. SO I WANT TO SHOW YOU WHAT CD8T CELLS ARE DOING TO THE VASCULAR WALL DURING THE DEVELOPMENT OF THIS DISEASE. IF WE GET RID OF THE CELLS WE CAN TAKE ANIMALS THAT HAVE FULL PATHOLOGY, BRAIN LEAK. THIS IS EVANS BLUE AND CONVERT IT OVER TO A CLEAN ASYMPTOMATIC MOUSE. THERE IS FULL SURVIVAL WHEN YOU REMOVE THE CD8T CELLS FROM THE SYSTEM. SO WHAT ARE THEY DOING? I CAN SHOW YOU THE MOVIES -- BUT THEY WOULD NOT BE VERY EXCITING. THEY DON'T DO ANYTHING LIKE YOU SAW IN THE FIRST FILMS FOR VIRAL MENINGITIS. WHAT WE CAUGHT WAS THIS SMOKING GUN FILM. WE'RE AT THE PEAK OF THE DISEASE WHEN THE MOUSE IS FULLY SYMPTOMATIC AND WE'RE TRACKING PARASITE SPECIFIC T CELLS AND WHAT WE CAN SEE IS THOSE CD8T CELLS ARE LINED UP ALONG THE VESSEL WALL AND THEY ARE STOPPING. THEY ARE ENGAGING THE SURFACE OF -- VASCULATURE DURING THE MAKE OF THIS DECEASE AND I HAVE NEVER SEEN ANYTHING LIKE THIS UNTIL NOW. -- SO IF WE GO INTO HUMANS AND SAY DOES THIS MATCH IN HUMAN BEINGS. WE DID A STUDY WITH SUE PIERCE'S LAB HERE AND WE WERE ABLE TO FIND MUCH THE SAME THING. WHEN WE LOOK IN THE VESSEL WALL OF HUMANS. CHILDREN IN AFRICA WHO HAVE SUCCUMBED TO THIS DISEASE. WE FIND THE CELLS ALONG THE VASCULATURE OF THE CHILDREN. WE BELIEVE THE MODE OF PATHOGENESIS IS IDENTICAL. SO WHAT HAPPENS TO THE VESSEL WALL? IT BASICALLY GETS VERY DISTRESSED AND AGAIN IS BEING ENGAGED BY CD8T CELLS. WE PUT IN A CALCIUM REPORTER. AND SO IN OUR STUDY STATE THIS IS A BLOOD VESSEL WITH NO DISEASE. THIS IS WHAT THE VESSEL WALL LOOKS LIKE. IT'S PASSING CALCIUM FROM CELL TO CELL AND WHEN WE HAVE A MOUSE THAT HAS SIGNIFICANT DISEASE WE SEE THIS PATTERN. AND THIS IS REALLY A STARK DIFFERENCE. YOU CAN SEE THESE VESSELS ARE NOW ENGORGED WITH CALCIUM. YOU CAN SEE THE FLUXES LIKE THIS VESSEL THAT PULSES WITH CALCIUM. YOU CAN SEE THE LEVEL IS WAY UP COMPARED TO THE PREVIOUS IMAGE THAT I SHOWED YOU. WHEN WE MONITOR THE CALCIUM DYNAMICS IN THE WALL YOU CAN SEE AGAIN IT GOES FROM A VERY QUIET BASAL FLUX DYNAMIC HERE TO WHAT WE SEE ON THE RIGHT AND THIS IS JUST HUGE DYNAMIC RANGE IN THE CALCIUM DYNAMICS SO THE BASIC QUESTION BECAME WHAT IS DRIVING THAT? THIS IS THE COMPARISON ON ART VEES VERSUS VEINS. WE GO FROM THE BLACK UP TO THIS DELTA OR THIS INCREASE IN FLUX ACTIVITY THAT YOU SEE IN THE ARTERIES AND THE VEINS OF THE SYMPTOMATIC MICE BUT WHAT CAUSES THIS ENDS UP BEING DIRECT ENGAGEMENT OF THE VESSEL WALL THAT ARE SPECIFIC TO THE PARASITE. HERE YOU CAN SEE A VERY CHAOTIC MOVIE. THE GOAL HERE WAS TO DELETE THE PRESENTATION FROM THE VASCULAR WALL AND SEE HOW THAT IMPACTED THE DISEASE PROCESS SO WE USED A VECTOR THAT SPECIFICALLY TARGETS THE VASCULAR WALL OF THE CNS AND THIS JUST SHOWS YOU THAT THIS VECTOR CAN GET INTO THE WALL QUITE NICELY AND THEN WE HAVE A MOUSE IN WHICH WE HAVE FLOX BETA TWO -- WE APPLY THIS VECTOR AND IT BASICALLY DELETES BETA TWO AND IT ONLY HAPPENS IN CNS VASCULATURE. AND WHEN WE DO THAT WE CAN PROTECT NEARLY ALL OF THE MICE FROM THIS DISEASE SO WHAT THAT TOLD US WAS THAT THOSE CELLS THAT I SHOWED YOU THAT ARE ENGAGING THE WALL NEED TO RECOGNIZE PEPTIDE IMAGE C ON THE WALL TO CAUSE THIS PARTICULAR DISEASE SO THAT WAS QUITE EXCITING. THE LAST PIECE OF DATA IS A THERAPEUTIC INTERVENTION. SO HOW DO WE TREAT TO STOP WHAT I JUST SHOWED YOU. WE DECIDED TO DO SOMETHING SIMPLISTIC. LET'S KNOCK THE CELLS OFF THE WALLACE THEY ARE ENGAGING THEM SO WE TREATED MICE WHEN THEY ARE BECOMING SYMPTOMATIC. DAY FIVE AND A HALF WITH ANTIBODIES. AND SO THOSE ARE THE ADHESION MOLECULES TO TETHER TO THE WALL AND WE ARE ABLE TO SAVE ABOUT 80% OF THE MICE FROM THIS PARTICULAR DISEASE AND THE MECHANISM IS CLEAN AND NOT THAT SOPHISTICATED. WE INJECT THE ANTIBODIES INTO THE BLOOD SUPPLY AND YOU CAN SEE THE T CELLS FALL OFF THE VESSEL WALL SO WE'VE CONVERTED AN ANIMAL THAT IS BEING DIRECTLY ENGAGED ON THE VESSEL WALL TO AN ANIMAL THAT IS NOT. WE CAN DO THIS ALMOST IN REALTIME. AND SO THE QUANTIFICATION OF THAT IS SHOWN HERE. WE GO FROM HAVING A MICE SYMPTOMATIC TO HAVING A MICE THAT HAS VERY FEW T CELLS ON THE WALL AND WE SAVE THEM FROM THIS PARTICULAR DISEASE. ON THAT NOTE I WILL CONCLUDE FOR MY ACKNOWLEDGMENT SLIDE THE PEOPLE THAT DID THE WORK WE HAVE A VERY NICE COLLABORATION WITH SUE PIERCES LAB AND WE GOT THE AAV VECTOR TO TARGET THE WALL FROM JACOB IN HAMBURG AND MOST OF THE STUFF THAT I SHOWED YOU -- WERE FROM MONICA AND PHILLIP. AND CAN CONCLUDE AND TAKE QUESTIONS LATER. >> THANK YOU VERY MUCH DORIANE. WE APOLOGIZE FOR TECHNICAL PROBLEMS. HOPEFULLY WE'RE BACK ON-LINE AND SHOULD BE FINE. OUR FOURTH SPEAKER OF THE DAY IS DR. CAROLYN ANDERSON -- AT THE UNIVERSITY OF MISSOURI. HER TALK IS ENTITLED -- IMAGING OF IMMUNE CELLS. >> SO HOPEFULLY YOU CAN ALL SEE MY SLIDES. THANK YOU SO MUCH DIMA FOR THE KIND INVITATION TO GIVE THIS TALK AND I'M ENJOYING THE OTHER TALKS AS WELL. SO WHAT I HOPE TO DO TODAY IS SHOW YOU A PROJECT THAT I WAS INVOLVED WITH AT THE UNIVERSITY OF PITTSBURGH WITH JOANNE FLYNN'S LAB AND WHAT WE ARE DOING IS LOOKING AT IMMUNE CELL IMAGING IN A MACAQUE MODEL OF TUBERCULOSIS. I MOVED TO THE UNIVERSITY OF MISSOURI AND WE ARE GETTING READY TO OPEN UP A SMALL ANIMAL IMAGING SUITE AT THE END OF THE YEAR AND WE HOPE TO BE CONTINUING SOME INFECTIOUS DISEASE WORK IN THAT FACILITY. SO NOBODY HAS ACTUALLY TALKED TOO MUCH ABOUT TUBERCULOSIS YET SO THIS IS A GOOD INTRODUCTION TO WHAT TUBERCULOSIS -- WHAT TYPES THERE ARE AND WHAT THEY CONSIST OF. GRANULOMA. I'M GOING TO GET A POINTER HERE. SO HERE WE HAVE THE MOST COMMON TYPE IS CACIAS. WE HAVE THIS CHEESE LIKE SUBSTANCE IN THE CENTER WHICH IS CALLED THE CASE YUM. AND THESE IMMUNE CELL -- COMPOSITION CAN TELL US SOMETHING ABOUT HOW AGGRESSIVE IT IS AND HOW IT MAY PROGRESS OR RESOLVE. SO HIGH NUMBER OF NEUTROPHILS CAN INDICATE WORSENING DISEASE. BUT AGAIN I THINK THERE IS STILL MORE WORK THAT NEEDS TO GO INTO THIS TO REALLY DEFINE THAT BETTER. SO THE FLYNN LAB PUBLISHED BACK IN 2014 AND THEY HAVE A MACAQUE MODEL OF TUBERCULOSIS WHERE THEY ARE ABLE TO INJECT THE MTB BACTERIUM INTO ONE OR BOTH LOBES OF THE LUNG AND IN COLLABORATION WITH A GROUP IN SOUTH KOREA AND ALSO A GROUP AT NAIAD. THIS RECAPITULATES HUMAN DISEASE AND THIS IS SHOWN WITH AND WITHOUT TREATMENT. AND THIS WAS DONE WITH FDG IMAGING WHICH SUCCESSFULLY AND READILY IMAGES TB GRANULOMAS. IT'S NOT EASY TO DISTINGUISH BETWEEN IMMUNE CELL POPULATIONS SO WE EMBARKED ON IMAGING ANTIGEN FOUR. WE THOUGHT THIS MIGHT BE MORE SPECIFIC FOR IMMUNE CELLS BUT IT'S ALSO EXPRESSED IN SOME EXTENT TO NEUTROPHILS AND T CELLS AND THESE CAN VARY. ANTIGEN FOUR IS -- IN TWO DIFFERENT ACTIVATION STATES SO THERE IS A LOW AFFINITY STATE AS WELL AS A HIGH AFFINITY STATE AND WE'RE SPECIFICALLY IMAGING THE HIGH AFFINITY STATE. WE HAVE A PEG LINKER BUT THE KEY -- WHICH HAS A 12.7 HALF LIFE. WE'VE BEEN DOING LOTS OF STUDIES. IT'S HIGHLY VERSATILE FOR BOTH CANCER SITUATIONS AND WE'VE DONE A LOT OF IT IN THE TB MODEL. WE ALSO PUBLISHED A PAPER THIS PAST YEAR IN SICKLE-CELL DISEASE. THIS COMPOUND IS VERY SPECIFIC FOR THAT HIGH AFFINITY ACTIVE FORM. AND THIS IS A PREVIOUS PAPER THAT WE PUBLISHED IN THE TB MODEL TO SHOW PROOF OF PRINCIPLE FOR THIS IMAGING AND HERE ARE TWO MACAQUE MODELS. AND WE UPON NECROPSY COLLECTED TISSUES AND COUNTED THEM AND YOU CAN SEE THAT WE'RE GETTING HIGHER UP TAKE AS ADVIS VISUALIZED. WE'RE ALSO GETTING HIGH UP TAKE IN LYMPH NODES. AND SO THAT WOULD SHOW THE PROOF OF PRINCIPLE THAT THIS TRACER WORKS QUITE WELL. AND JOSH DOES THIS VERY PAINSTAKING WORK WHERE HE LOOKED AT GRANULOMAS THAT HAD HIGH INTAKE AND LOW INTAKE. HE DID THIS FOR VARIOUS MARKERS SHOWING THAT AND THEN HE COUNTED CELLS AND CORRELATED THIS WITH THE CPMs AND YOU CAN SEE HERE THAT THE MACRO FACIAS TEND TO TAKE UP MORE THAN THE T CELLS OR -- CELLS OR NEUTROPHILS. 8 SO IN THIS PARTICULAR STUDY *. WE WANTED TO LOOK AT COPPER -- VERSUS FDG AND IN MACAQUE MODEL WHERE WE WERE TREATING WITH -- WE HAVE A PET/CT SCANNER AND BSL3 AT THE UNIVERSITY OF PITTSBURGH AND WE IMAGED WITH FDG A FEW TIMES TO MONITOR WHETHER WE HAD INFECTION. WE STARTED A BASELINE SCAN WITH A COPPER -- AT WEEK 12 AND THEN WE FROM WEEKS 15-21 WE WERE TREATING WEEKLY OR DAILY WITH -- AND ALTERNATING -- IMAGES WEEKLY AND THEN WE MACAQUE MODELS WERE SACRIFICED AND NECROPSIES DONE AT WEEK 21. THIS IS HOW WE PRODUCE THE TRACER. IT'S VERY STRAIGHTFORWARD PRODUCTION OF THE TRACER. WE DID DO A TEST ON THAT FOR -- AND THEN WE INJECTED ABOUT 8-10 MILL CURRIES AND WE DID THE PET IMAGING AROUND FOUR HOURS POST INJECTION. WE ALSO HAD THE CT WHICH WAS VERY NICE TO HAVE. WE WERE ABLE TO THEN LOOK AT EACH INDIVIDUAL GRANULOMA AND WE COULD GET SIZE AND MEASUREMENTS FROM THE CT. WE DID A WHOLE PET HOT WHERE WE WENT SLICE BY SLICE THROUGH THE LUNG AND GET THE SUV THROUGHOUT EACH SLICE AND WE ALSO DID THE INDIVIDUAL GRANULOMA ANALYSIS THAT GAVE US SIZE AS WELL AS THE SUV DATA FROM THE PET WHICH IS THE STAND-UP TAKE VALUE. THIS IS A SLICE. HERE IS THE FDG IMAGING. THEY CAN PICK UP THE GRANULOMA BUT YOU DO GET SLIGHTLY DIFFERENT UP TAKE FOR EACH INDIVIDUAL TRACER. HERE IS THE TOTAL ANALYSIS AND THIS IS SORT OF A CONGLOMERATION OF THE CONTROLS -- HAD HIGHER PET HOT FOR EACH TRACER COMPARED. THE THIS IS WHEN YOU LOOK AT ALL THE SLICES TOGETHER COMPARED TO THE TREATED. WE HAD FOUR TREATED ANIMALS AND WE HAVE TWO UNTREATED IN THIS GROUP THAT I'M GOING TO PRESENT TO YOU TODAY. BUT I THINK WHAT I'M GOING TO SHOW YOU IS THAT WE MAY SEE SOME -- THIS ONE LOOKS LIKE A NONRESPONDER BUT THE PET HOT DOESN'T ALWAYS TELL THE WHOLE STORY. SO HERE IS THE INDIVIDUAL GRANULOMA ANALYSIS AND LOTS OF DATA ON THIS. THIS IS THE FIRST TWO SUBJECTS I'M GOING TO SHOW YOU ARE THE UNTREATED ONES. AND THESE EACH -- POINT HERE IS AN INDIVIDUAL GRANULOMA AND THE WAY THAT I DID THE DATA ANALYSIS WAS TO LOOK AT THE PERCENT CHANGE FROM THE BASELINE SCAN. SO YOU CAN SEE HERE THAT EACH INDIVIDUAL GRANULOMA EITHER THE SUV VALUE CHANGES AT WEEKS 2 AND WEEK 6. SAME FOR THE LPO2A AND THIS IS THE SIZE BASED ON THE SCANS AND AGAIN THEY COULD CHANGE SLIGHTLY ON WHEN THE IMAGING OCCURRED. BUT YOU CAN SEE THAT THE UP TAKE DOESN'T NECESSARILY CORRELATE WELL. YOU HAVE SOMETIMES YOU HAVE HIGH UP TAKE AND YOU DON'T SEE A BIG CHANGE IN THE SIZE OF THE GRANULOMA AND THERE ALSO IS SOME HETEROGENEITY. SO THESE ARE THE TWO UNTREATED SUBJECTS. HERE IS TWO OF THE TREATED SUBJECTS. AGAIN -- THE FDG AND LOP2A AND SUBJECT 217 DOES LOOK LIKE THE UP TAKE IS INCREASING. THIS ONE LOOKS MORE LIKE A RESPONDER. THIS IS SUBJECT 221. AND THEN THE CHANGE IN GRANULOMA SIZE TELLS SOMETHING A BIT DIFFERENT AND IT LOOKS LIKE 217 IS A MIXED RESPONDER AND NOT A NECESSARILY A NONRESPONDER. AND THEN THESE ARE THE OTHER TWO TREATED SUBJECTS AGAIN. YOU CAN SEE THERE IS ABSOLUTELY HETEROGENEITY BETWEEN EACH GRANULOMA AND WE DID NOT GET A TWO WEEK SCAN DUE TO TECHNICAL ISSUES BUT YOU CAN STILL SEE THE CHANGE FROM BASELINE OUT TO SIX WEEKS IN SUBJECT 224 AND IN 234 LOOKS LIKE A PRETTY GOOD RESPONDER. BUT WE KEY INCREASED UP TAKE. JUST TO SHOW YOU WHETHER IS THERE A CORRELATION AND THESE ARE AGAIN THE SAME GRANULOMAS AND SOMEWHAT LOSE CORRELATION BUT I WOULD NOT SAY ANYTHING GREAT AND SO THERE ARE LIKELY IMAGING IMMUNE CELL POPULATIONS. HERE ARE SOME EXAMPLES OF THE IMAGES THAT GO ALONG WITH THE DATA. THESE ARE 3-D PROJECTIONS AND WE DID NOT LOOK AT THE LYMPH NODES AT THIS POINTS BUT WE WILL BE ANALYZING THOSE DATA AS WELL. HERE IS SUBJECT 220 WHICH IS UNTREATED SUBJECT. AND YOU CAN SEE THAT THE -- THIS ONE HAS ACTIVE LUNG DISEASE THAT INCREASES OVER TIME AND FROM NECROPSY ONE OF THE GRANULOMAS THAT WAS REMOVED. HERE IS A TREATED RESPONDER. AGAIN THE GRANULOMAS DECREASED IN SIZE. AND IN THIS CASE ALMOST ALL OF THE LUNG LESIONS THEY DEFINITELY SHRUNK. AGAIN THE LLP2A SHOWS MORE SIGNAL THAN THE FDG IN THIS CASE AND HERE IS AN EXAMPLE OF EXTENSIVE SCARRING OF ONE OF THE GRANULOMAS SHOWING THAT IS RESOLVING AND THIS ONE WHICH AGAIN THE PET HOT SUGGESTS AND EVEN SOME OF THE DATA SHOWS THAT IT'S A NONRESPONDER BUT IF YOU LOOK AT THE SIZES IT'S A MIXED RESPONDER AND IN THIS PARTICULAR CASE THE NECROPSY SHOWED THAT THEY HAVE A LOT OF THE FOAMY MACRO FACIAS * PRESENT AND THAT MAY BE THE RESULT OF THE HIGHEST UP TAKE IN THIS PARTICULAR GRANULOMA AND YOU CAN SEE THAT SOME OF THE -- ARE SOME RESOLVING AND ARE SOME NOT. SO WE ALSO LOOKED AT CFUs AND THE TOTA TOTE -- THORACIC -- THIS IS A PROCEDURE THAT THE FLYNN LAB HAD DEVELOPED. JUST A ROUGH ESTIMATE. SO IF WE WANT TO COMPARE PRE AND POST WHAT WE DEFINED IS POST NECROPSY WAS THAT THE DRUG TREATED SUBJECTS HAD DECREASES IN FCF USA COMPARED TO THE CONTROLLED SUBJECTS. * SO SEEN THE 217 WAS DEFINITELY A RESPONDER. MOSTLY A RESPONDER. SO-CON CLUED WE CAN LOOK AT DISEASE CONTROL AND PROGRESSION. WE CAN MONITOR IT DYNAMICALLY. THERE IS HETEROGENEITY OBSERVED FOR BOTH PET TRACERS THAT DOES NOT NECESSARILY CORRELATE WITH TREATMENT RESPONSE. THE LLP2A AND FDG UP TAKE LOOSELY CORRELATE WITH EACH OTHER. THIS IS A STUDY STILL IN PROGRESS. WE'VE COLLECTED ALL OF THE DATA NOW. WE HAVE ALL OF THE NECROPSIES. WE'RE LOOKING AT CORRELATING INDIVIDUAL GRANULOMA ANALYSIS -- AND WE ALSO HAVE SIX MORMON KEYS SCANNED WITH DATA YET TO BE ANALYZED. SO WE HAVE 8 TREATED AND FOUR CONTROLLED. AND SO I WANT TO ACKNOWLEDGE THE COLLABORATION I HAD WITH THE FLYNN LAB SINCE WHEN I ARRIVED AT THE UNIVERSITY OF PETS IN 2011 UNTIL I LEFT JULY OF LAST YEAR AND WE'RE STILL WORKING CLOSELY TOGETHER. WE STILL HAVE PAPERS THAT WE'RE WORKING ON I'VE LEARNED A TREMENDOUS AMOUNT FROM -- THIS WAS NOT MY INITIAL AREA OF PET IMAGING AND DEVELOPING TRACERS BUT IT HAS BEEN VERY PRODUCTIVE AND REWARDING EXPERIENCE TO WORK WITH THIS GROUP OF PEOPLE. MIKE IS ONE OF THE GRAD STUDENTS FROM PIT WHO IN THE BIOENGINEERING PROGRAM I CONSIDER HIM STILL PART OF MY LAB AND I'M STILL ONE OF HIS MENTORS SO HE IS CONTINUING TO DO SOME DATA ANALYSIS WITH JOSH ON THIS PROJECT SO THAT WE CAN FINISH THIS UP AND MAYBE HOPEFULLY GET A BETTER PICTURE OF WHAT IS GOING ON THAN WHAT I SHOWED YOU TODAY SO WITH THAT I'LL -- THANK YOU ALL AND I LOOK FORWARD TO QUESTIONS DURING THE DISCUSSION. 7. >> THANK YOU VERY MUCH CAROLYN. THIS WAS A WONDERFUL TALK. I WOULD LIKE NOW TO ASK ALL OF THE SPEAKERS TO HAVE THEIR CAMERAS ON. DR. ANDERSON, MURMUR, -- McGavern AND HUGHES. >> -- THE FIRST QUESTION. -- COULD THESE BE POTENTIALLY USED -- -- >> THANK YOU VERY MUCH FOR THIS VERY GOOD QUESTION. 7 SO I WOULD LIKE TO ANSWER THIS GENERALLY. BECAUSE WITH IMAGING YOU HAVE GOT ALWAYS THE PROBLEM THAT YOU REALLY DON'T KNOW ARE YOU IMAGING A GOOD ANTIGEN OR A LOT OF CELLS AND TO DISCRIMINATE THE ANTIGEN EXPRESSION AND THE NUMBER OF CELLS YOU PROBABLY NEED VERY CHALLENGING STUDIES THAT COMPARE THESE DIFFERENT STRATEGIES TO IMAGE IMMUNE CELLS AND THIS IS CHALLENGING BUT IT'S A PROMISING OUTLOOK TO IMAGE THE FUNCTIONS OF DIFFERENT LINES OF T CELLS AND IMAGING THE LEVEL FOR EXAMPLE -- EXPRESSION WHICH ARE MUCH MORE VIABLE THAN THE CD3 EXPRESSION THAT WE USED BUT THIS IS DEFINITELY THE PERSPECTIVE YOU WANT TO GO TO. >> THANK YOU VERY MUCH. SECOND QUESTION IS FOR DR. HUGHES. HOW FAR EASY IS LABELING AND WHAT KIND OF REACTIVE IS BEING USED AND WILL YOU BE WILLING TO SHARE WITH OTHER INVESTIGATORS ON A COLLABORATIVE BASES. WE JUST GET PURIFIED ANTIBODIES. YOU NEED QUITE A LOT. SO IT CAN GET COSTLY. SORRY. IT'S LATE. I STARTED WITH THE LAST ONE FIRST. WHAT TYPE OF RADIOACTIVITY IS USED. >> HOW EASY IS LABELING. >> HOW EASY IS IT TO LABEL THE COLON? >> ZIRCONIUM. >> I DIDN'T DO IT. YOU WOULD HAVE TO ASK PEOPLE IN THE LAB -- WE MAKE IT IN HOUSE. IT WAS JUST A STRAIGHT CONGREGATION AND I THINK IT WAS EASY ENOUGH THAT NICOLE WAS DOING A PhD AND SHE MANAGED TO GET THAT DONE. SO I THINK IT'S FAIRLY STRAIGHTFORWARD. BUT I'M NOT A CHEMIST. >> WE HAVE SIMILAR EXPERIENCES. VERY RELIABLE LABELING METHODS THAT CAN BE LEARNED BY ANYBODY. >> YEAH. >> GREAT AND I THINK I FORGOT THE LAST ONE AS WELL. WAS THERE ANOTHER ONE? >> WHAT KIND OF RADIOACTIVITY IS BEING USED? AGAIN. >> THERE WAS ANOTHER QUESTION ACTUALLY -- ABOUT SO THIS IS A QUESTION FOR DORIANE. -- LABELING TECHNIQUES ARE VERY ENJOYABLE TO WATCH. I LIKE YOUR FASHION WHEN YOU SPEAK. GREAT LECTURE. EXCELLENT TALK. HOW LONG CAN YOU IMAGE THE ANIMALS. ALSO DOES IT AFFECT BLOOD LEVEL PHYSIOLOGY. >> IN TERMS OF IMAGING TIME THAT DEPENDS ON THE STATE OF THE ANIMAL IN TERMS OF DISEASE. IF YOU TAKE A NAIVE ANIMAL AND SET UP AN IMAGING EXPERIMENT YOU COULD PROBABLY IMAGE FOR 10-12 HOURS STRAIGHT. THAT IS A LONG IMAGING EXPERIMENT. ON AVERAGE WE PROBABLY IMAGE FOR ONE TO TWO HOURS. WHEN YOU IMAGE MICE THAT ARE SICK -- I WOULD SAY IT'S HARD PRESSED TO GET AN ANIMAL IMAGED FOR MORE THAN ONE HOUR BECAUSE THEY ARE SYMPTOMATIC AND COMPLICATIONS. AND THEY TEND TO HAVE ISSUES JUST HANDLING ANESTHESIA AND STAYING UNDER WHILE YOU'RE IMAGING THEM SO THE STATE OF THE MOUSE MAKES A HUGE DIFFERENCE. THE OTHER QUESTION WAS ABOUT THE ANESTHESIA. IT CERTAINLY INFLUENCES BLOOD VESSEL AND FLOW. THE USE OF CAT MEAN WILL SLOW THE HEART RATE DOWN * SO YOU'LL HAVE A DECREASE IN BLOOD PRESSURE. BY CONTRACT ISO FLORINE WOULD NOT HAVE THAT SAME IMPACT SO LIKELY SOME INFLUENCE ON BLOOD FLOW BASED ON WHETHER YOU SAY KEPT MINOR * ISO FOR YOUR ANESTHESIA. >> THANK YOU. QUESTION FOR DR. ANDERSON. -- -- >> ABSOLUTELY AND IN FACT MY COLLABORATOR AT THE UNIVERSITY OF PITTSBURGH JUST PUBLISHED A PAPER IN ACUTE LUNG INJURY. NOT SURE IT'S IN THE JOURNAL BUT DEFINITELY ON -- IT WAS ACTUALLY PUBLISHED IN FEBRUARY OR MARCH. SO I THINK THAT CERTAINLY INFLAMMATORY BOWEL DISEASE WOULD BE VERY INTERESTING. I REALLY ENJOYED THE TALK AND NOW I'M HAVING A TOUGH TIME REMEMBERING WHO GAVE IT. I THINK IT WAS PATRICK ABOUT LOOKING AT THE MOD OOOH, LATORS VERSUS THE IMMUNE * CELLS AND ALSO LOOKING AT FIBROSIS VERSUS INFLAMMATION AND I THINK -- BETA -- IS PROBABLY NOT GOING TO BE AS GOOD FOR FIBROSIS. FOR CHRONIC CONDITIONS DEFINITELY NOT BUT FOR MORE ACUTE I THINK IT WOULD BE GREAT. >> MAY I ADD TO THAT. >> GO AHEAD. >> TARGETING WITH -- IS ONE OF THE BETTER DRUGS TO COME OUT. WHAT WE SEE IS ABOUT 20% TO 40% IS PRIMARY RESPONDERS. WE DON'T KNOW WHY THERE IS SUCH A BAD UP TAKE AND THAT IS WHERE THIS IMAGING CAN HELP. IF I STAYED IN ACADEMIA THAT IS WHAT I WANTED TO DO. >> SO AS LONG AS WE HAVE A MINUTE HERE IS THAT THE ALPHA 4 BETA 7 IS THAT DRUG AN ANTIBODY OR A SMALL DRUG. >> ANTIBODY. IT'S OUT ON THE MARKET. SO YOU CAN GET SOME. >> ANOTHER QUESTION FOR PATRICK AND NEIGHBOR EVERYBODY ELSE AS WELL. WITH THE INCREASED -- INTESTINAL INFLAMMATION -- AND INTERPRETATION OF THE RESULTS. >> POSSIBLY. POSSIBLY. YOU'VE GOT LEAKY BLOOD VESSELS. THE TRACER COULD BE LEAKING OUT OF BLOOD VESSELS RATHER THAN ACCUMULATING. YEAH. THAT IS A TRICKY ONE TO ANSWER BECAUSE THE LEAKY BLOOD VESSELS ARE ALSO THERE WITH INFLATION AND IMMUNE CELLS AS WELL. THAT IS HARD. >> JUST GOT ANOTHER ONE. ZIRCONIUM ANTIBODIES OFTEN HAVE NONSPECIFIC BOWEL UP TAKE. >> BOWEL UP TAKE. THAT IS WHY WE DID THE UP TAKE SO YOU CAN SEE IT'S CLEARLY CHANGED FROM THE INFLAMMATORY AND ALSO IN THE FIBROSIS MODEL AS WELL. >> AS A CONTROL. OKAY. WE DON'T HAVE ANY MORE QUESTIONS FROM VIDEO CAST. DO YOU HAVE ANY QUESTIONS -- FOR EACH OTHER OR ANYBODY ELSE? >> OKAY. PERFECT. >> I HAVE A QUICK QUESTION -- SINCE MOSTLY THE GROUP IS DOING A LOT OF LABELING WITH RADIO METALS AND ANTIBODIES THE FIRST QUESTION IS ABOUT THE ANTIBODIES. WHEN YOU DO THE REPEAT IMAGING WITH THE ANTIBODIES IS IT POSSIBLE THAT ALL THE ANTIBODIES DEVELOP AND HOW FREQUENTLY ARE PEEING E PEOPLE SEEING THIS? BECAUSE THAT IS ONE OF THE LIMITATIONS THAT PEOPLE BRING UP WHEN YOU'RE USING ANTIBODY IMAGING THAT THE BODY WILL FORM ANTIBODIES AGAINST IT AND IT WILL NOT BE AS EFFECTIVE IN THE FUTURE OR MAY CAUSE REACTIONS. IS THAT TRUE OR ARE PEOPLE OBSERVING THAT IN REPEAT IMAGING? >> SO MAYBE I'M ANSWERING -- WE DID NOT OBSERVE IT IN OUR MODELS BUT THIS CANNOT BE BECAUSE THE MOUSE ARE NOT HAVING THEIR OWN IMMUNE SYSTEM. THE ANNUAL BODY WE USED WAS A COMMERCIALLY APPROVED DRUG WHICH WAS ALSO APPLIED IN PATIENTS TO SUPPRESS IMMUNE RESPONSE AND TRANSPORTATION AND IT WAS OBSERVED IN SOME PATIENTS THERE ARE ANTIBODIES AGAINST THIS MOUSE ANTIBODIES AND THIS COULD BE TYPICALLY OVERCOME WITH -- ANTIBODIES THAT HAS GOT A HUMAN BACKBONE AND IT'S NOT REALLY A PROBLEM WITH TYPICALLY PROVED ANTIBODIES BECAUSE THEY ARE FROM HUMAN OR AGAIN. -- ORIGIN. >> THANK YOU. >> ONE MORE QUESTION FOR CAROLYN. COUNTY GROUP DISCUSS RADIO METAL KEY LACKS. * MAYBE DR. ANDERSON WOULD BE BEST TO ANSWER THIS. >> SURE. I THINK WITH INTACT ANTIBODIES WHICH THERE WAS PRESENTATIONS TODAY ZIRCONIUM IS QUITE SIMPLE. IT'S IN THE LITERATURE ALL OVER THE PLACE. THERE ARE NEW KEY LATORS COMING OUT * THAT PEOPLE ARE LOOKING AT TO GIVE SOME MORE VERSATILITY. FOR THE SMALL MOLECULE THAT I PRESENTED THE LPA2 -- GAVE BETTER IMAGES AND THE 12 HOUR SLICE IS NICE. IT'S NOT THERE -- WHEN WE DO WEEKLY THERE IS NO CROSS -- BUT GETTING A GOOD COMPOUND WOULD BE GREAT. IT'S BEEN A LITTLE BIT ELUSIVE. AND AS FAR AS KEY LATORS GO * THERE IS A LOT OUT THERE SO I THINK THAT IS A LONG ANSWER SO I WON'T GO INTO IT BUT THERE ARE DEFINITELY GOOD KEY LATORS FOR ALL OF THESE RADIO METALS PRESENTED IN THE LITERATURE AND COMMERCIALLY AVAILABLE. >> THANKS A LOT, CAROLYN. WE HAVE TIME TO TAKE ONE MORE QUESTION. THIS IS FOR DORIANE. ONCE YOU STOP THE CELLS IN MALARIA HOW IS THE INFECTION CLEARED? >> THAT'S AN IMPORTANT QUESTION AND I DIDN'T HAVE A CHANCE TO ADDRESS THAT. THE GOOD NEWS IS THAT THE PATHOLOGY I SHOWED YOU WITH THE BLOOD BRAIN BARRIER IS NOT ACTUAL CLEARANCE OF THE INFECTION. IT'S AN ACCIDENT. SO WHAT'S IS THE INFECTION IS IN THE RED BLOOD CELL AND THERE IS SOME TRANSFERENCE OF MATERIAL TO THE VESSEL WALL AND THE -- CROSS PRESENT THAT ANTIGEN AND CAUSE THE T-CELLS TO ATTACK IT BUT THERE IS NO ACTUAL PARASITE IN THE -- ENDO FILLIAN -- *. THEY ARE NOT NEEDED TO CLEAR THIS PARTICULAR PARASITE. YOU CAN CLEAR IT MUCH MORE EFFECTIVELY WITH ANTIBODY AND THE ANTIBODY WOULD ATTACK THE RED BLOOD CELL WHICH IS WHERE THE PARASITE IS. SO THE ANTIBODY WOULD BE NECESSARY TO REMOVE THE INFECTION. THE CELL -- WE'RE INHIBITING AS THIS ACCIDENT AND SAVING THIS PATIENT FROM THE FATAL DISEASE SO ANTIBODIES ARE THE KEY. >> THANK YOU VERY MUCH EVERYONE. THOSE WERE WONDERFUL TALKS AND WE'RE GOING TO CONCLUDE OUR SESSION. WE'RE GOING TO TAKE A BREAK FOR 10 MINUTES AND WE SHOULD BE BACK AT 10:20 FOR OUR SECOND SESSION OF THE DAY. WELCOME BACK. I WILL BE MODERATING THIS SESSION. WE NOW HAVE FOUR WONDERFUL SPEAKERS TALKING WITH US SHORTLY. I WANT TO MAKE A QUICK REMINDER THAT THESE SPEAKERS WILL PRESENT THEIR WORK CONTINUOUSLY WITHOUT BREAKS. SO IF YOU HAVE ANY QUESTIONS USE THE FEEDBACK OPTION ON YOUR VIDEO CAST SCREEN. AND AT THE END OF THE SESSION WE WILL HAVE SPEAKERS BACK TO ANSWER ALL OF YOUR QUESTIONS. WITH THAT IT IS MY GREAT PLEASURE TO INTRODUCE OUR FIRST SPEAKER DR. SANJAY JAIN. >> THANK YOU VERY MUCH. IT'S WONDERFUL TO BE TALKING AT THIS MEETING. MANY THANKS TO EVERYBODY FOR ORGANIZING AND SPEARHEADING THIS. I'M GOING TO CHAT A LITTLE BIT ABOUT OUR EXPERIENCE WITH DEVELOPING BACTERIA SPECIFIC IMAGING OR PATHOGEN SPECIFIC IMAGING AND I WANT TO ECHO WHAT WAS SAID THIS MORNING. IT'S EASY TO TALK WITH THE AUDIENCE HERE THAT INFECTIONS ARE IMPORTANT. EVEN IN 2017 INFECTIONS RANKED THIRD IN MORTALITY BUT FIRST IN MORBIDITY. AND THERE ARE A FEW IMPORTANT PATHOGENESIS INCLUDING BACTERIAL PATHOGENESIS BUT ALSO -- THAT ARE CAUSING DISEASE GLOBALLY. AND THIS IS JUST THE COST IN THE U.S. -- THE ANNUAL COST DIRECT MEDICAL COST THIS DOES NOT INCLUDE THE IN DIRECT COST BECAUSE OF MISSED TIME AT WORK ETC. THIS IS THE COST OF HEALTH CARE ASSOCIATED INFECTIONS. AND AS WAS POINTED OUT -- RESISTANT INFECTIONS WILL BECOME THE LEADING CAUSE OF DEATH IN 2050. $100 TRILLION DUE TO DRUG-RESISTANT INFECTIONS AND I DON'T KNOW HAHNEMANY TRILLIONS OF HAVE BEEN LOST DUE TO COVID. IT'S IMPORTANT TO THINK ABOUT ARE WE USING THE BEST TECHNOLOGIES WE HAVE IN INFECTIOUS DISEASES. I BRING UP THE EXAMPLE OF TB. THAT IS HOW WE STARTED IN INFECTIOUS DISEASE IMAGING AND THE FUNDAMENTAL DIAGNOSTICS IS THE MICROSCOPE AND THAT REMAINS SO. BUT WE HAVE FANCIER METHODS BUT WE STILL NEED A CLINICAL ISOLATOR. AND THAT MEANS THAT WE NEED TO GO AND ISOLATE THE BUG WHICH OBVIOUSLY CAN BE INVASIVE SO THE PROBLEM IS WE NEED AN INVASIVE BIOPSY OR WE HAVE TO GO IN AND GET THEM WITH A NEEDLE OR SURGERY. OFTEN WHEN WE'RE TRYING TO GET AT A LESION THAT WE CAN SEE -- AND THEN WE SCHEDULED A BIOPSY AND SOMETIMES WE MISS IT BECAUSE IT'S A SMALL LESION AND NOT ACCESSIBLE. THERE IS ALSO PET AGAIN NAME TEE. -- HETEROGENEITY. * IT COULD BE LIKE CANCER -- INFECTIONS CAN METASTASIZE. IT'S DANGEROUS AND GENERALLY LIMITED TO A SINGLE TIME POINT. WE CANNOT DO BIOPSIES AGAIN AND AGAIN. YOU CAN BUT IT'S QUITE DIFFICULT. I'M A BIG FAN OF STAR TREK. IT CAN IMAGE WHAT IS HAPPENING AND ALSO FIX SOME NICKELS. BLOOD VESSELS. WE HAVE IMAGING MACHINES AND YOU ALL KNOW THE BASIC PRINCIPLES OF THE RADIO PHARMACEUTICAL IMAGING AND THAT IS WHAT WE FOCUS ON AND THERE IS A LOT OF INVESTMENT AND THE QUESTION IS CAN WE ACTUALLY USE THESE TECHNOLOGIES FOR INFECTIOUS DISEASES. I DO WANT TO POINT OUT THIS GRAPH AND THIS IS THE NUMBER OF PUBLICATIONS NOT ASYMPTOMATIC ANALYSIS BUT A QUICK ANALYSIS ON PubMed. AND IN THE LAST COUPLE OF YEARS THERE HAS BEEN A SPIKE IN THIS AND THAT IS THANKS TO ALL OF YOU AND I THINK IT'S VERY IMPORTANT TO MAINTAIN THE MOMENTUM. I DON'T THINK EVERY TRACER THAT WE DEVELOP IS GOING TO GO INTO THE CLINIC OR BE CLINICALLY USED BUT EVEN IF A FEW OF THEM ARE THAT WILL BE BRILLIANT. SO QUICKLY I WANT TO DESCRIBE A LITTLE BIT OF THE PROGRAM THAT WAS DEVELOPED IN OUR LAB ON DEVELOPING PATHOGEN SPECIFIC IMAGING FOR BACTERIA WITH THE HYPOTHESIS THAT SMALL MOLECULES -- COULD BE UTILIZED AS BACTERIA SPECIFIC IMAGING. I'M GOING TO SHOW YOU SOME DATA AND SHOW YOU HOW WE TRANSLATE SOME OF THE STUFF FROM THE PRECLINICAL DISCOVERY -- AND THE CLINIC. AND THIS IS THE WORK OF A LOT OF PEOPLE. I WANT TO TALK BRIEFLY -- THIS IS PUBLISHED IT WAS A PUBLICATION BY SEVERAL MEMBERS FROM THE SUB GROUP THAT YOU HAVE TO BE CAREFUL ABOUT HOW YOU CHOOSE YOUR TARGET AND TRACER BECAUSE REALLY TO BE ABLE TO DIFFERENTIATE A BACTERIAL INFECTION FROM THE SURROUNDING TISSUE YOU REALLY NEED A LOT OF TRACER TO BE EITHER CUMULATED OR BIND TO THE TARGET WHICH IS THE BACTERIA BECAUSE THEY ARE MUCH SMALLER COMPARED TO THE -- CELLS WHICH IS THE HUMAN CELLS. SO YOU EITHER NEED TO HAVE MULTIPLE BINDING OR SOME APPLICATION. SO THE TRACER -- WORKED THIS IS A COUPLE OF YEARS AGO. THIS STARTED IN 2011-12. AND WE SCREENED ABOUT 1,000 AND WE ASKED THREE QUESTIONS. WE WANTED TO LOOK FOR SPECIFICITY AND ACCUMULATION AND WE WANTED TO KNOW THE -- METABOLISM. I'M GOING TO SHOW YOU A TABLE. WHAT WE FOUND WAS THE MOLECULES THAT SCREENED THE HIGHEST PLUS 3 WHICH MEANS MOST LIKELY CANDIDATES THAT WOULD BE DEVELOPED -- WE BROUGHT THOSE TRACERS AGAINST VARIOUS SPECIES TO SEE IF IT WOULD BE TAKEN UP BY THEM. ONE WAS TAKEN UP BY ALL BACTERIUM AND THERE WERE CERTAIN OTHERS THAT WERE TAKEN UP SELECTIVELY BY GROUPS OR CLASSES OF BACTERIA AND THAT IS IMPORTANT AND I'LL DISCUSS A LITTLE BIT ABOUT THAT IN THE NEXT FEW MINUTES. WE'RE LUCKY WITH ONE CALLED SORBITOL. -- WE'VE DEVELOPED TWO FLOOR OH PABA AND I'M GOING TO TALK ABOUT A TRACER THAT WE'VE TAKEN INTO THE CLINIC. BRIEFLY ABOUT PABA THE PABA WORK HAS NOW GONE INTO ANIMALS AND WE HAVE MORE EXTENSIVE DATA. THIS IS SOME WORK THAT -- HAS BEEN DOING WHERE THERE IS A JOINT INFECTION MODEL IN RABBITS AND YOU CAN SEE NICELY THAT PABA -- ACID CAN LIGHT IT UP. WE ALSO HAVE DONE -- IMAGING UNTIL HEALTHY VOLUNTEERS SHOWING THERE IS LOW BACKGROUND ESPECIALLY IN THE LUNGS. SO FIVE FEMALES AND WE'VE GOTTEN RADIATION -- AND IT'S VERY FAVORABLE. SO I WANT TO TALK A LITTLE BIT ABOUT THE FDS STORY. AND I THINK THE BEST THING IS IT CAN BE MADE EASY. THIS IS A MOUSE THAT HAS AN INFECTION. YOU CAN SEE THE YELLOW ARROW AND DEAD BACTERIA ON THE OTHER SIDE. AND YOU CAN SEE THAT FDS WAS INTERJECTED AND YOU CAN SEE THE PIE THAT HAS LIVE BACTERIA LIGHTS UP NICELY BUT THE DEAD BACTERIA DOES NOT AND THERE IS VERY LOW BACKGROUND OTHERWISE. THIS GI UPDATE -- AND THE URINE BLADDER HAS BEEN CATH RISED. * AND THIS IS TWO HOURS LATER SO THE KIDNEYS ARE NOT LIGHTING UP AS MUCH. THIS IS THE FDG -- AND YOU CAN SEE THAT FDG CANNOT DISTINGUISH BETWEEN LIVE AND DEAD BACTERIA. WE ALSO WANT TO POINT OUT THAT YOU CAN DIFFERENTIATE AND QUICKLY FIGURE OUT IF INFECTION IS BEING TREATED APPROPRIATELY. THIS IS A DRUG SUSCEPTIBLE E. COLI. ON THE LEFT SIDE THIS IS PRETREATMENT. ON THE RIGHT SIDE AFTER 24 HOURS OF ANTIBIOTICS. THE CT LOOKS ALMOST THE SAME AND THAT IS ACOD BY A CHANGE IN THE BACTERIA BURDEN. BUT IF YOU GIVE THEM AN ANTIBIOTIC THAT DOES NOT WORK THAT DOES NOT HAPPEN. THAT IS THE BASES FOR A CLINICAL STUDY. AND I'M NOT GOING TO GO THROUGH A LOT OF DETAILS EXCEPT THAT IT WAS EXTENSIVE WORK THAT TOOK A LOT OF YEARS. BECAUSE WE USED STRINGENT CRITERIA TO DO THIS CLINICAL STUDY. SO IN THIS STUDY WE WERE TRYING TO UNDERSTAND IF FDS PET WOULD BE ABLE TO PICK UP INFECTIONS BECAUSE OF A GROUP OF BACTERIA THAT IS TARGETED. AVENUE ALSO TO BE ABLE TO FIND OUT IF IT IS SPECIFIC ENOUGH OR NOT WHICH MEANS THAT OTHER ONCOLOGIES LIKE CANCER OR INFLAMMATORY DISEASES THAT DON'T HAVE INFECTIONS WOULD THEY BE PICKED UP OR NOT. THE CLINICAL DEFINITIONS WERE STRINGENT. -- SO OBVIOUSLY WE WOULD NOT DO THIS IF IT WAS NOT CLINICALLY INDICATED SO ONLY A SELECT FEW PATIENTS WERE FULFILLING THIS CRITERIA AND THAT INFECTIONS PEOPLE WITHOUT INFECTION -- PEOPLE WITH SUSPECTED INFECTION BUT WHERE THE CONFIRMATION WAS NOT THERE THEY WERE EXCLUDED FROM THE STUDY AND THEN WE HAD CONTROLS. I WANT TO SHOW YOU SOME DATA FROM THE STUDY. THIS WORK WAS DONE IN COLLABORATION WITH A HOSPITAL IN COLUMBIA AND THIS IS ACTUALLY A PATIENT WHERE THERE IS A 60-YEAR-OLD MAN WITH LUNG DISEASE WITHOUT INFECTION. YOU CAN SEE SOME UP TAKE. THERE IS SOME DISEASE IN THE LUNG BUT YOU DON'T SEE ANY FDS UP TAKE. THIS IS A PATIENT THAT HAS -- ALSO PNEUMONIA AND YOU CAN SEE A LOT OF SIGNAL IN THE LUNG. AND THIS WAS OBVIOUSLY CONFIRMED -- BY -- AND IF YOU LOOK AT TWO-HOUR IMAGES NOW IF YOU SWITCH BACK TO THE LEFT SIDE OF THE PANEL YOU CAN SEE THAT THE LIVER UP TAKE IS LOWER BUT WE DON'T SEE ANY UP TAKE IN THE LUNG WHERE THERE IS SOME DISEASE AND IN THIS PATIENT ALSO YOU CAN SEE SOME MAX IN THE LUNG BUT AND YOU CAN SEE SOME SIGNAL ON THE LEFT SIDE OF THE PATIENT. SO TUNE IN TO THIS PATIENT BECAUSE I WANT TO SHOW YOU THAT THEY ARE PICKING UP DIFFERENT --. I HOPE TO SHOW YOU A SLIDE. THESE ARE SOME OTHER PATIENTS. THAT ALSO WAS BECAUSE OF E. COLI AND YOU CAN SEE THOSE. THE OTHER THING IS THAT IN THIS MODEL IN THIS STUDY THE PEOPLE WHO HAD BACTERIAL INFECTIONS HAD A MUCH HIGHER -- -- AND WE SET UP A CUT OFF OF THREE. WE HAD ABOUT 82% WITH 100% -- WE SET A CUT OFF OF THE RATIO. SO DECENT CHARACTERISTICS AND WE NEED ADDITIONAL LARGER STUDIES. THIS PAPER IS PUBLISHED. THERE ARE SOME LIMITATIONS BUT YOU SHOULD ALSO LOOK AT. THIS IS THE PATIENT THAT I WAS TALKING ABOUT. THIS PATIENT GOT AN FDG AND THE FDS FOR RESEARCH REASONS. AND YOU CAN SEE THE PANEL ON THE RIGHT SIDE YOU CAN SEE MASS IN THE LUNG ON THE RIGHT LUNG WHICH WE THINK AR ARE MEDS. THE LAST THING THAT I WANT TO SHOW YOU IS TREATMENT RESPONSE. SO WHEN THESE PEOPLE WERE TREATED AND THE STUDY TEAM WAS NOT MAKING THE DECISIONS ABOUT WHO IS RESPONDING TO TREATMENT OR NOT. THIS WAS A CLINICAL DECISION BUT IN THE PATIENTS WHO RESPONDED TO TREATMENT THERE WAS A DECREASE A SUBSTANTIAL DECREASE IN THE TARGET TO NONTAR INDICATE RATIO FOR F DS. SO JUST SOME DATA HERE. TO SHOW THAT. ONE QUICK THING I WANT TO ADD -- PHILLIP WHO'S A RADIO CHEMIST WHO HAS DEVELOPED A VERY INTERESTING METHOD WHERE YOU CAN MAKE FDS -- FROM FDG WITHIN ABOUT 10 MINUTES AT ROOM TEMPERATURE SO WE THINK THAT THIS WOULD REALLY HELP EXPAND THE USE OF FDS BECAUSE ANY PLACE THAT HAS FDG CAN MAKE FDS AT ROOM TEMPERATURE. AND OBVIOUSLY PHILLIP CAN TALK MORE ABOUT THAT. ALSO SOME INTERESTING EXPERIMENTS WHEN HE TRIED TO SHOW -- PNEUMONIA -- IN PATIENTS WHO ARE ADMITTED WITH COVID -- THAT THIS WAS IN A HAMSTER MODEL THAT FDS COULD DIFFERENTIATE THIS. ON THE TOP YOU'RE SEEING FDG PET AND YOU CAN SEE THAT THE MOUSE. BUT ON THE RIGHT PANEL YOU SEE FDS PET AND YOU SEE NO SIGNAL EVEN THOUGH THERE IS A LOT OF DISEASE IN THE LUNG BECAUSE OF SARS CO-V2. AND THE FDS PET WITH THE KNEW MAIN. >> INFECTION IS SIGNIFICANTLY HIGHER THAN THE F DS PET BECAUSE OF COVID 2. YOU CAN READ ABOUT IT. I WANT TO TELL YOU ANOTHER INTERESTING THING. THEY'VE UTILIZED THE ABILITY OF ALL OF THESE TRACERS TO GET OUT OF THE KIDNEYS RAPIDLY BECAUSE THEY ARE NOT ME THABOLIZED BY THE HUMAN BODY. *. SO I KIND OF WANT TO TELL YOU -- THIS IS A SIDE PROJECT BUT REALLY NICE TO SHOW HOW IT WORKS WELL. JUST TO SUMMARIZE BACTERIAL INFECTIONS OR IN GENERAL REMAIN A THREAT TO HUMAN HEALTH AND WE NEED BETTER TOOLS TO BE ABLE TO MONITOR AND DIAGNOSE THEM AND WE NEED TO BE ABLE TO DIFFERENTIATE INFECTION FROM NONINFECTIOUS PROCESSES. AND THEN WE NEED INFORMATION ON SPECIFIC MONITORING OF RESPONSE TO TREATMENT AND THERE ARE SEVERAL TRACERS THAT ARE BEING DEVELOPED. PABA IS BUT YOU'LL HEAR ABOUT OTHER TRACES THAT ARE BEING DEVELOPED. I TALKED ABOUT THE FDS WHERE WE'VE DONE A CLINICAL STUDY FOR DETECTION OF -- AN OLDER TERM. AND THERE ARE A COUPLE OF GOOD THINGS ABOUT FDS. EASILY MADE AND THE FIRST -- LOOKS PROMISING. ULTIMATELY WE CAN ALSO USE A LOT OF THIS FOR DISEASE PATHOGENESIS. I WANT TO THANK ALL OF THE PEOPLE IN MY LAB WHO HAVE DONE THIS WORK. PLEASE WHISPER THAT ALVARO IS LOOKING FOR A RESIDENCY. BUT ALL OF THE COLLABORATORS OVER THE LAST FEW YEARS THAT HAVE REALLY DONE AMAZING WORK WITH US, I ALSO WANT TO THANK THE NIH THAT HAS FUNDED ME. SO THANK YOU VERY MUCH AND I GUESS WE PROBABLY WILL GO TO THE NEXT SPEAKER. 7 >> THANK YOU, DR. JAIN. WITHOUT FURTHER DELAY I WOULD LIKE TO INTRODUCE OUR NEXT SPEAKER DR. DAVID WILSON. HIS TOPIC IS IMAGING INFECTION USING BACTERIA SPECIFIC METABOLIC PATHWAYS. >> GREAT. I HOPE EVERYBODY CAN HEAR ME THAT I'M NOT MUTED OR OTHERWISE COMPROMISE ON THIS LECTURE. THANKS FOR THAT GREAT INTRODUCTION SANJAY. I TOOK MY CARBON 11 PABA STAR TREK SLIDE OUT OF THIS SLIDE DECK BECAUSE SEVERAL OF THE THINGS THAT WE'RE TRYING TO DO WITH PABA -- * THEY BELONG IN THE REALM OF STAR WARS AT LEAST FOR NOW. THE AREAS ARE REALLY DEVELOPMENT IN MR BASED. WE SEEM TO HAVE BETTER LUCK GETTING FUNDED -- BUT THEY ARE BOTH OF HIGH INTEREST. SO I'M GOING TO TALK ABOUT MY MOTIVATION. WHICH HAS TO DO WITH A CHALLENGING CLINICAL AND DISCITIS-OSTEOMYELITIS. THIS IS A DIAGNOSIS THAT IS INCREASINGLY IMPORTANT. PEOPLE ARE USING MORE IV DRUGS AND ARE GETTING MORE DISKITIS. THIS IS A SLAM DUNK DIAGNOSIS THAT WE SEE FREQUENTLY UNFORTUNATELY. BUT THIS IS WHAT WE SEE ALSO FREQUENTLY IS CASES THAT ARE DJD MIMIC. THEY CAN REALLY BE THE SAME WHEN RUN OF THE MILL DEGENERATIVE DISEASE WHICH WE ALL HAVE VERSUS ACUTE INFECTION. IT WOULD BE GOOD TO HAVE SENSITIVE TRACERS IN THIS AREA. THE NUCLEAR IMAGING TOOLS THAT HAVE BEEN APPLIED TO THESE. ACTIVATED IMMUNE CELLS AND TO A LESSER EXTENT BACTERIA LIKE TO TAKE UP GLUCOSE. ACT LIKE CANCER AND THAT IS WHY WE'RE REASONABLY SENSITIVE TO PICKING THEM UP WITH FDG ALSO LESS SPECIFIC. THE MAIN ISSUE IS LACK OF SPECIFICITY. SO WHAT HAVE WE DONE TO LOOK AT THIS? SANJAY INTRODUCED THIS FANTASTIC WORK. THIS IS ONE REASON WHY I'M WORKING IN THIS FIELD. TO REPURPOSE IT FOR DRAMATIC RESULTS THAT ARE PUBLISHED NOW TWICE MOST RECENTLY WITH AN OUTSTANDING CLINICAL DATA. WE'VE BEEN THINKING ABOUT THE APPLICATION FOR SEVERAL YEARS. IF WE'RE GOING OVER DISKITIS. IT'S AT LEAST 90% OF OUR BACTERIA -- I'M RELATIVELY OBSESSED WITH THIS. WOULD LOVE TO SEE WHAT THE BEST F18 TRACER WE HAVE TO DO THIS. BUT WE STARTED WITH THIS IDEA THAT AMINO ACIDS WERE AN INTERESTING WAY OF ACCOMPLISHING THIS BECAUSE THEY HAVE SUCH A LARGE PERCENTAGE OF PEPTO -- THIS MIGHT BE A GOOD WAY OF DETECTING THEM AND THERE IS A COUPLE OF WAYS OF DOING THIS SO WE THOUGHT ABOUT -- THIS SUPREME JANNA MEAN OH ACID. AND WE * WE THOUGHT ABOUT OTHER D AMINO ACIDS THAT WILL ACTIVELY ACTIVE ACCUMULATED. SO YOU CAN PUT IN D AMINO ACIDS THAT WILL BE SWAPPED. AND SO THERE ARE TWO POTENTIAL MECHANISMS OF IN CORPORATION THAT WERE IN POSITION TO INVESTIGATE MORE FULLY. BUT WE STARTED OUT SANJAY MENTIONED THIS ARTICLE. KIND OF PROBLEMATIC BECAUSE YOU CANNOT SCREEN YOUR NEW COMPOUNDS AND YOU'RE STUCK WITH WHAT IS COMMERCIALLY AVAILABLE. BUT WE STARTED WITH THIS IDEA THAT THEY COULD DETECT BACTERIA AND GOT SOME BIG HITS. AND DID SOME INVITRO WORK. AND ENDED UP BY A PRETTY BIG RATIO. AND THIS SHOWED A PROMISING INITIAL RESULTS. BACKGROUND IS HIGHER THAN SOME OF THE DATA. YOU COULD BE BOTH CO-LIE AND STAPH ORIENTED. WE STARTED TO LOOK AT THIS MORE CAREFULLY. SO DEVELOPED A RAPID PROTOCOL FOR SCREENING ALL SORTS OF STUFF SIMULTANEOUSLY. NOW THROWING THESE TRACERS INTO 12 ORGANISMS IN PARALLEL TO LOOK AT THEIR RELATIVE UP TAKE AND FINALLY DEVELOPING AN AUTOMATED SYNTHESIS THAT IS BASED ON THE LINEAR -- AS OPPOSED TO A CYCLICAL -- SO WE'VE BEEN PUTTING THIS INTO PATIENTS. WE'VE STUDIED THREE MEN HEALTHY -- THREE WOMEN HEALTHY AND THREE POTENTIALLY INFECTED PATIENTS. THIS IS DIFFICULT BECAUSE WE'RE WORKING WITH CARBON 11 AND OUR PET/CT SCANNERS -- IN AN OUTPATIENT FACILITY. IT'S EXTREMELY HARD TO GET THESE PATIENTS THAT YOU WANT TO IMAGE SO WE'VE BEEN IMAGING THESE POTENTIALLY INFECTED PATIENTS THAT WE'VE OBTAINED FROM OUR ORTHOPEDIC SURGERY COLLABORATIONS AND THE DATA CERTAINLY LOOKS INTERESTING. WE HAVE TO DIGEST IT. WE'VE BEEN SEEING ASYMMETRIC UP TAKE IN SOME OF THESE PATIENTS THAT ARE INFECTED OR HAVE PROSTHETIC JOINT INFECTIONS SO WE HAVE TO LOOK AT PATIENTS THAT ARE OBVIOUSLY INFECTED AND FIGURE OUT WHAT IS TO DISSEMINATE THIS TECHNOLOGY. PART IS LOOKING FOR SOMETHING THAT IS F18 SO WE CAN TRANSPORT IT AND ACCESS THE PATIENT SCANNERS. THERE IS GENERALLY THIS PROBLEM IN PET WHICH IS FOR ACTIVELY INFECTED PATIENTS OF COURSE YOU WANT THEM TO BE SCANNED AS INPATIENT BUT HAVING THE SCANNER IS GENERALLY VERY INCONVENIENT FOR ONCOLOGY APPLICATION. SO HOW ABOUT CARBON D. THIS IS MOST PRESENT D AMINO ACID. FROM THIS LINEAR -- GLUE SEEN PRECURB CERTIFY. AND BOTH OF THEM WORKED WITH JUST THE -- SHOWING HIGHER SENSITIVITY AND PROBABLY THE Y AXIS IS THE MOST OBVIOUS THING TO TAKE AWAY. THEY TEND TO BE FIVE OR TENFOLD. THEY ARE NOT -- THE D VERSUS L DIFFERENCE IS DRAMATIC BETWEEN THE WAY THEY TAKE THESE UP AND THESE SHOW WHEN YOU PUT THIS TRACER INTO AN -- WITH BACTERIUM IN IT AND LET IT INCUBATE IT THERE WILL BE MORE RADIOACTIVITY IN THE PELLET -- SO EXTREMELY SENSITIVE. THIS IS SHOWN THAT WE COULD DO THIS VERY SENSITIVE AND SPECIFIC DETECTION OF BACTERIA IN THIS DUAL INFECTION MODEL. DATA RECAPITULATES THAT FDG CAN'T DO THIS WHEREAS ALOMINE IS VERY GOOD AT DOING THIS. 7 SO WE'VE SEEN THAT RESULT MANY TIMES. OF COURSE THIS CAN DETECT ANTIBIOTIC. STARTED WITH RESISTANCE VERSUS SENSITIVE. AND THIS WAS REALLY COOL -- WAS LOOKING AT ADVANCED MODELS. WE'VE DONE THIS -- AND NOW THROWN ABOUT 7 TRACERS INTO THIS MODEL SO INTO A STAPH ORIOUS RAT. AND OPTICAL IMAGING BUT LIGHT UP INCREDIBLY WITH D -- ALOMINE. THE BEST IS PROBABLY PABA. BUT ANY WAY I THINK IT'S EXTREMELY PROMISING. A LOT OF THESE TRACERS FOR BETTER ELUCIDATION. WE'VE SEEN SOMETHING SIMILAR AND ARE GOING TO FULLY INVESTIGATE THE EFFECT. JUST A PIECE OF DATA. I DON'T KNOW IF OTHER PEOPLE WORKING IN THE FIELD HAVE THE SAME PROBLEM BUT I DON'T THINK I'VE HAD A GRANT COME BACK WITH FAILURE TO DISCUSS AND GIVE US A HARD TIME ABOUT THE MICROBIOME AND IS THE BACKGROUND YOU SOAKED DUE TO BACKGROUND METABOLISM OR THE MICROBIOME. SO WE'VE PRESENTED FROM GOOD EVIDENCE BY SHOWING THESE THAT DON'T HAVE A MICROBIOME. SO THIS WAS MAYBE A GOOD WAY TO ANSWER IS THAT QUESTION ABOUT WHAT THE NORMAL MICROBIOME IS SHOWING. WHAT ARE WE GOING TO DO WITH THIS? WE'RE IN THE BROTHER SESSION. WE PERFORMED AN AUTOMATION TO GET THAT TRACER INTO PEOPLE. WE WANT TO LOOK AT BIOFILM MODELS. F18 DERIVATIVES -- FROM THE PUBLISHED WORK THAT THEY ARE PROMISCUOUS WITH DETECTION OF BACTERIA. HOWEVER SOME MODIFICATIONS WILL STILL A BLAKE THE ABILITY TO ENCORP NATE THE ACID DERIVATIVES. THIS IA LOT OF THESE THINGS -- I THINK LIKE I PRESENTED THIS SLIDE WITH A LITTLE -- CRYING EMOJI HERE. SO FAR OUR WORK HAS NOT RECAPITULATES THE SENSITIVITY OF THE CARBON TRACERS. HOW TO MAKE CF3. YOU CAN DO THIS POTENTIALLY WITH -- GLUE SEEN WITH A CF3 TYPE GROUP. -- THIS IS PRETTY INTERESTING. SO I THINK LIKE THERE IS THIS PROBLEM WITH THESE PROBE THAT'S WE'RE IDENTIFYING WHICH IS THE MORE SENSITIVE THE RADIO TRACER IS FOR BACTERIA THE MORE BACKGROUND WE GET. MY STRONG SUSPENSION IS THAT DAL LEAN IN MAMMALS -- IT'S PROBABLY * BEING OXIDIZED AND MAKING PIROVATE AND THEN YOU HAVE SOME BACKGROUND AT THE CROSSROADS OF BIOCHEMISTRY. WE'VE BEEN THINKING ABOUT USING DIFFERENT ICE TOPPERS * TO ABLATE THAT BACKGROUND SIGNAL SO, THAT IS SOMETHING THAT WE'RE TRYING VERY HARD TO DO CURRENTLY. AND THEN MOVING ON TO MR BRIEFLY. THIS IS A REALLY FASCINATING IDEA. WE WOULD LOVE TO DO MRI IN PEDIATRIC POPULATIONS WITH NONIONIZING TOOLS. WE WOULD LOVE TO HAVE THE SPECIFICITY. WE HAVE THIS TECHNOLOGY THAT WE'VE BEEN WORKING ON OCTAVELY THAT YOU INTRO INTRODUCE HYPERPOLARIZED SUBSTRATE A AND IT MAKES SOMETHING IN REALTIME F. WE GET PATIENTS WITH PROSTATE CANCER WHERE THE SIGNAL IS RAMPED UP IT MAKES LACTATE SO ALL YOU SEE IS LACTATE IN TUMORS. IF YOU GIVE THIS TO THE NORMAL HEART IT GETS DEBOX LATE. AND ACTUALLY YOU SEE BUY CARBONATE THAT IS RELATED TO CO2. IF YOU GIVE IT TO BACTERIA WE SAW THIS SIGNAL IN AS SAME. WE'VE GIVEN A -- THERE ARE TWO PARADIGMS. SOME METABOLIZES BY BACTERIA ONLY. SOME THAT HAVE BEEN DEVELOPED DO THIS AND THE SECOND IS SUB STRAIGHT GENERATES BACTERIA SPECIFIC METABOLITE. THE FIRST INVOLVES PET AND THE SECOND CONSIDERING PIPER POLARIZED C13 SO WE HAVE THESE BACTERIA GROWING IN THIS TUBE THAT ARE PROFUSED AND ENCAPSULATED AND THEY MAKE A TON OF ACETATE AS WE'VE SHOWN AND WE'RE THINKING ABOUT HOW TO MAKE AN AS STATE SPECIFIC IMAGE OF THOSE DATA SO IF A PATIENT HAS A BACTERIAL INFECTION AND YOU GIVE THEM PIRATE THE ONLY THING MAKING ACETATE WILL BE THE BACTERIA. SO CAN WE ACTUALLY IMAGE THIS? THERE ARE A LOT OF OBSTACLES BUT AN INTERESTING AND PROMISING IDEA. WE HAVE THIS MRI IDEA. THIS HA A LOT OF IDEAS. YOU STILL HAVE THE SPECIFICITY OF CHEMICAL SHIFT. IT'S CERTAINLY TRUE THAT YOU HAVE THIS CONVERGE END -- TRANSFORMATION OF SORE TO THE MAN TO THE. YOU MAKE SOME MORE BUT YOU CAN SENSE BOTH SO IF YOU'RE MAKING LACTATE OR ETHANOL YOU'RE SEEING THE BACTERIA. SO WE'RE WORKING ON METHODS TO DETECT THIS. THIS IS PUBLISHED IN 1988. YOU PUT IN GLUCOSE INTO BACTERIA AND THIS IS EVERY MINUTE UP TO A COUPLE OF HOURS. IF YOU PUT IN THIS SIX POSITION LABELED GLUCOSE YOU GET OUT THIS LACTATE AND ETHANOL QUANTITATIVELY FROM THE BACTERIA THAT IS GROWING IN CULTURE. WHAT IS A COOL RESULT. SO JUST TO CONCLUDE THESE TECHNOLOGIES ARE JUST GREAT FOR METABOLIC IMAGING. WE LOVE TO WORK IN CARBON 11. IT'S KIND OF LIKE THIS ADDICTION WHERE OUR PROJECTS DON'T WORK AS WELL WHEN WE TRY TO MOVE TOWARD F18 BUT WE'RE GETTING THERE SLOWLY. THIS STATEMENT IS KIND OF TRUE. THE MOST POWERFUL -- -- I WOULD LOVE TO HEAR ABOUT AND I'M NOT SURE WHO HAS THIS TRACER. MAYBE IT'S STANFORD. BUT I WOULD LIKE TO SEE A TRANSLATAL PET TRACER THAT IS F18 LABELED AND ACCUMULATED BY ALL GRAM NEGATIVES AND POSITIVES I THINK WOULD BE A GOOD STARTING POINT FOR ADDITIONAL TRANSLATION OF THIS TECHNOLOGY AND OF COURSE THE SPECIFICITY IS AN INTERESTING QUESTION AS WELL. WITH THAT I'LL THANK YOU FOR YOUR ATTENTION. THANK ALL OF THESE PEOPLE IN MY GREENED AND FUNDING THAT NEEDS TO BE UPDATED. AND I THINK THAT WE'RE HOLDING COLDS SO PERHAPS LATER. SO THANK YOU VERY MUCH. >> THANK YOU, DR. WILSON. CERTAINLY A VERY INFORMATIVE AND SET OF INFORMATION THAT YOU HAD THERE. OUR NEXT IS DR. XIANKA SUN. >> CAN YOU SEE MY SLIDES AND HEAR MY VOICE? >> YES. >> VERY GOOD. THANK YOU. THANK YOU FOR THE INTRODUCTION AND I ALSO I WOULD LIKE TO GIVE MY THANKS TO -- FOR THEIR INVITATION AND ALSO THE ORGANIZER OF THIS SYMPOSIUM FOR SUCH THE WONDERFUL OPPORTUNITY. I THINK IT'S -- FOR US ESPECIALLY AS A NEW COMER TO THE FIELD. TO I'M GOING TO TALK ABOUT OUR WORK ON COMPARATIVE EVALUATION OF D VERSUS L GLUTAMINE. IMAGING OF BACTERIAL INFECTIONS. SO SANJAY HAD PRESENTED THE BACKGROUND INFORMATION. A BACKGROUND OF THE IMAGING OF BACTERIAL INFECTION -- AND -- [ INDISCERNIBLE ] AND WE KNOW IT'S PROJECTED BY 2050 THAT THE DRUG RESISTANT INFECTIONS ARE GOING TO OVERTAKE CANCER TO BECOME THE LEADING CAUSE OF DEATHS GLOBALLY. SO CLEARLY -- MANAGE A PATIENT WITH INFECTIONS. >> DR. SUN I'M SORRY TO INTERRUPT BUT CAN YOU GET CLOSER TO THE MIC. >> IS THAT BETTER? >> YEAH. I THINK THIS IS BETTER. >> OKAY. I'M SORRY. SO TRADITIONALLY -- WHEN INFECTION IS AFFECTED AND -- THE FIRST STEP AND THE PATIENT'S OF SYMPTOMS AND THEN WHEN THAT TASK IS PERFORMED -- -- FROM INFECTION SITE. SO -- IN ADDITION TO THE BIOPSY RISK AS WELL -- -- -- -- SOME TIMES ESPECIALLY IN CASE LIKE THE INFECTION SITE IS UNKNOWN. AND TRADITIONAL DIAGNOSTIC METHODS CAN BE CHALLENGING OR DIFFICULT AND HARD TO PERFORM. -- IT HAS THE POTENTIAL TO REVOLUTIONIZE THE DIAGNOSIS AND ALSO THE TREATMENT RESPONSE -- FOR INFECTIOUS DISEASES. THIS IS BECAUSE OF MOLECULAR IMAGING. CAN BE PERFORMED OVER THE WHOLE BODY OF THE PATIENT AND -- IT CAN BE PERFORMED NONINVASIVELY AND AS WELL AS -- HOWEVER TO DO SUCH SCANS ESPECIALLY FOR PET WE NEED TO HAVE BACTERIAL SPECIFIC AGENTS. AND TODAY THANKS TO THE PIONEER WORK AND ACTUALLY MANY OF THE WORK THAT WAS REPORTED BY THE TEAM OF TODAY'S SPEAKERS IN SO MANY HAVE MADE POSSIBLE TO -- AND SOME HAS BEEN CLINICAL TRIALS AND SANJAY PRESENTED -- -- IN CLINICAL TRIALS. AND OUR WORK -- IN THE CATEGORY OF -- SO GENERALLY SPEAKING TO DESIGN OR BACTERIA SPECIFIC IMAGING AGENTS WOULD HAVE TO FOCUS ON THE INTRINSIC DIFFERENCE OF THE BACTERIA. THE GUIDING PRINCIPLE IS TO IDENTIFY TARGET FROM BACTERIA SPECIFIC -- BIOCLINICAL MARKERS OR -- PATHWAY. SO -- BACTERIAL INFECTION -- IN DR. WILSON JUST SHOWED WAS SAYING -- FROM HIS PAPER -- USING -- IT'S ACTUALLY -- THIS IS BECAUSE WHEN AND THE ALGORITHM PRESENT ONLY BACTERIA USED AS D AMINO ACIDS AND ALSO BUILDING MATERIALS TO CONSTRUCT -- SO AS YOU CAN SEE FROM HERE AND THE MANY -- [ INDISCERNIBLE ] DR. WILSON JUST MENTIONED -- FOR BACTERIA INFECTION IMAGING IT'S VERY PROMISING. WHAT I WANT TO POINT OUT HERE IS IN THIS CARTOON -- THIS IS -- GLUTAMINE -- AND SOME PUBLISHED WORK IT WAS REPORTED AS YOU CAN SEE HERE THE PATIENT -- IN THE -- BACTERIAL CELL SYNTHESIS WAS IN THE FORM OF ISO -- ACID BUT FOR US IT DOESN'T MATTER BECAUSE WE INCORPORATE IT -- AND. [ INDISCERNIBLE ] WE SEE COME FROM THE BACTERIAL WALL. THIS SUMMARIZES THE BACTERIAL GROWTH -- ONE THING I WANTED TO POINT OUT IS -- THEY CONDITION CONVERTED IN VIVO JUST BY -- RESPECTFULLY. AND IF. [ INDISCERNIBLE ] TO TRACE THIS METABOLIC PATHWAYS. HOWEVER -- THERE IS NO SUCH -- [ INDISCERNIBLE ] OR RESHOULDN'T SEE SOM -- [ INDISCERNIBLE ] -- BASED ON THIS RATIONAL WE THINK -- CAN BE USED OR CAN BE DEVELOPED TO DEVELOP FOR SPECIFIC BACTERIAL IMAGING. SO FIRST -- LET'S TALK ABOUT RADIO SYNTHESIS OF GLUTAMINE. AND ACTUALLY * WE HAVE IN THE CLINICAL PRODUCTION OF -- WE JUST NEED TO SWAP THE -- PRECURSOR AND THE REST -- IS IDENTICAL. AND SO FAR -- AUTOMATE THE -- [ INDISCERNIBLE ] BOTH D AND L TRACERS. OVER 35% -- AND OVER 90%. SO. [ INDISCERNIBLE ] WE BASI BASICALLY HAVE MADE D -- AVAILABLE FOR STUDY. WE FIND OUT OUR ENTIRE METHODS AND WE SAW -- A FEW MINUTES DIFFERENCE BETWEEN D AND L GLUTAMINE. SO THIS IS BASICALLY INDICATES THAT WE DON'T HAVE TO USE THE INITIAL -- FOR THE RADIO SYNTHESIS. TO MAKE D AND L GLUTAMINE -- [ INDISCERNIBLE ] TO SEPARATE D FROM THE L GLUTAMINE BETA TRACERS. AND WE USE POSITIVE BACTERIA -- [ INDISCERNIBLE ] -- TO EVALUATE THE TIME DEPENDENT UP TAKE WE -- BACTERIA WITH THE RADIO TRACERS AND GLUTAMINE DEFICIENT. AND ESPECIALLY -- [ INDISCERNIBLE ] HERE I'M SHOWING YOU THE -- RESULTS AND AS EXPECTED -- SHOWING A MUCH HIGHER UP TAKE OF L GLUTAMINE AND THEN D GLUTAMINE GLUTAMINE. AND WHEN WE LOOK CLOSER AND ACTUALLY THE UP TAKE OF RATE OF E. COLI IS MUCH HIGHER AS COMPARED TO -- SO THIS IS ACTUALLY A VERY NICE FEATURE WHEREAS WE CAN SEE THE IMAGING -- [ INDISCERNIBLE ] AND THE INTERESTING -- WE FOUND OUT DIFFERENT BACTERIA STRING HAD DIFFERENT USE OF DIFFERENT DEGREE OF USE OF D AND L GLUTAMINES SO THIS MAKES US WONDER IF WE CAN USE -- PET IMAGING PERFORMED -- WITH D AND L GLUTAMINE FOR SPECIFIC BACTERIA STRING IDENTIFICATION. AND HERE IS THE COMPARING FOR THE UP TAKE -- -- [ INDISCERNIBLE ] INCREASED UP TAKE OF L GLUTAMINE AND ACTUALLY FROM 30 MINUTES TO -- 90 MINUTES WE SEE 8 FOLD INCREASE. HOWEVER FOR D GLUTAMINE -- WE SEE SUCH -- OF UP TAKE. SO BECAUSE OF THE RESULT AND -- [ INDISCERNIBLE ] AND WE PERFORMED COMPARATIVE IMAGE STUDY USING D AND L -- GLUTAMINE AND WE DID A STUDY IN MICE TO GET A STEADY WE HAVE A BASELINE SCAN. AND AFTER THAT WE TREATED THE MICE WITH TWO DOSES OF -- TO MAKE THE ANIMALS -- SO WE CAN ESTABLISH -- [ INDISCERNIBLE ] INFECTIONS IN THE MICE. [ INDISCERNIBLE ] AND SHOWED -- INJECTION FOR THE CONTROL. -- WAS AFTER THE INFECTION SO WE DID -- A 20 MINUTE SCAN WITH BOTH D AND L. AFTER IMAGING WE HAVE SOME TISSUE -- [ INDISCERNIBLE ] AND ALSO THE TISSUE FOR HISTOLOGY. SO I'M SHOWING YOU A COMPARATIVE -- AND AS YOU CAN SEE FROM THE -- A MUCHER HIGHER INTAKE OF THE -- TISSUES AND ACTUALLY EXCEPT FOR KIDNEY OTHER ORGANS SHOWED A HIGH UP TAKE -- ACTUALLY IT'S WHAT WE EXPECT. BECAUSE ANIMALS THE HOST -- THE ATTRIBUTES OF -- AND WE SEE A -- BACKGROUND AND THE KEY UP TAKE OF GLUTAMINE IS MUCH HIGHER. THIS IS INDICATED -- [ INDISCERNIBLE ] FOR KIDNEY AND EXCHRETIEN. AND AS A RESULT WE COULD SEE A HIGHER CONTRAST. [ INDISCERNIBLE ] THE CONTRAST IS SIMPLY WAS OF THE BACKGROUND. SO THE CONTRAST IS BASICALLY -- ROUGHLY ABOUT ONE. AND FOR D THE CONTRAST THE BACKGROUND IS -- SO WE SAY HIGH CONTRAST. SO, THAT IS THE MAIN REASON -- WE SEE -- [ INDISCERNIBLE ] D GLUTAMINE. AND TO FULLY CONFIRM THE SPECIFICITY OF BACTERIA -- WE PERFORMED A COMPARATIVE STUDY -- -- THE MOUSE MODEL AS YOU KNOW -- [ INDISCERNIBLE ] -- BACTERIAL INFECTION. AS YOU CAN SEE HERE -- FDG SHOWED A HIGH CONTRAST IN THE -- AND GLUTAMINE BASICALLY NOTHING SO THIS IS A FULL INDICATOR THAT GLUTAMINE CAN BE USED FOR BACTERIAL SPECIMEN IMAGE. SO HERE IS -- HISTOLOGY AND UNCOVERED -- [ INDISCERNIBLE ] AND CLEARLY THE RESULTS CONFIRMED -- OUR IMAGE RESULTS SO IN -- INDICATED -- -- BOTH GRAM POSITIVE AND GRAM NEGATIVE. AND BACTERIAL -- IMAGING IS -- HAS A HIGHER CONTRAST AND SPECIFICITY THAN L GLUTAMINE AND IN CONCLUSION -- PET IMAGING -- [ INDISCERNIBLE ] IS CAPABLE AND DIFFERENTIATING ACTIVE BACTERIAL INFECTION FROM STERILE INFLAMMATION IN MAMMALIAN HOSTESS. 7 -- A COLLABORATION BETWEEN SOUTHWESTERN AND UT ARLINGTON AND THE GRADUATED STUDY SUPPORTED BY UT SOUTHWESTERN -- -- AND GRANTS FROM -- CANCER PREVENTION AND RESEARCH INSTITUTE OF TEXAS AND THIS HAS NOT BEEN FUNDED BY EXTENDED FUNDING SOURCE AND -- WE'RE WORKING HARD TO GET A GRANT TO SUSTAIN OUR FUTURE WORK. I WOULD LIKE TO THANK EVERYBODY FOR YOUR TIME AND ATTENTION. >> THANK YOU, DR. SUN. OUR FINAL SPEAKER IS DR. YONG LI. A POSTDOC RESEARCHER AT STONEY BROOK UNIVERSITY AND HIS TALK IS ENTITLED IMAGING BACTERIAL INFECTION WITH PRODUCTION OF F18F-PABA. >> THANK YOU FOR THE NICE INTRODUCTION. SO REALLY TODAY MY TALK I WILL TALK ABOUT SOME OF THE WORK AND EFFORTS OF IMAGING BACTERIAL INFECTIONS WITH METABOLITE TRACERS. OUR WORK IS BASED ON THE SIGNIFICANT WORK OF DR. JAY'S LAB IN TERMS OF ACTIVITY AND METABOLIZED SCREENING EFFORTS THAT LED TO PABA BASED TRACERS AND THEN BASED ON THAT WE STARTED WITH FLORINE ENABLED PABA BUT I WILL TALK BRIEFLY ABOUT THAT AND THEN I'LL MOVE MOST OF MY TALK. >>> -- THE EFFORTS THAT WE DID TO IMPROVE THE BEHAVIOR ASSOCIATED WITH THIS FIRST GENERATION F PABA. AS WE KNOW THE -- ESPECIALLY INDUCED IS A CLEARLY AMONG MEDICAL LEADS. AS YOU CAN SEE IN THE PICTURES -- THOSE INFECTIONS CAN CAUSE VERY HIGH MORTALITY AND BASED ON THE STATISTICS CLEARLY THESE ARE VERY DANGEROUS INFECTIONS IN THE CLINIC AND PART OF THE REASONS WHY SO HIGH MORTALITY ASSOCIATED WITH THESE INFECTIONS IS BECAUSE WE'RE LACKING OF A SIGNIFICANT DIAGNOSIS METHOD. -- THE METHOD THAT WE'RE USING IS THE ULTRASOUND AND THAT IS BASED ON THE STRUCTURE CHANGES ASSOCIATED WITH INFECTIONS WHICH ALWAYS HAPPENS AT LATER STAGES OF THE INFECTION SO THIS IS ONE OF THE REASON WHY SOME OF THE -- CANNOT CAPTURE THE EARLY INFECTION EVENT. AVENUE FOR THE GOAL TO -- DETECTION WE HEAVILY RELY ON SOME OF THE TISSUE SPECIMENS WHICH IS SOMETIMES VERY DIFFICULT TO OBTAIN. 8 AND LAST BUT NOT LEAST WE'RE LOOKING A METHOD THAT CAN PROVIDE EARLY PHARMACODYNAMIC BIOMARKER FOR THE CHELATION TO ASSESS IF THE TREATMENT IS EFFECTIVE AT THE EARLY STAGES. BASED ON THIS WE'RE KEEN TO LOOKING FOR TECHNOLOGIES THAT CAN BRIDGE THE GAP OF DETECTING THESE CLINICAL RELEVANT DISEASE INFECTIONS. SO ONE -- WHICH IS VERY PROMISING IS THE -- MOLECULAR IMAGING. IT DETECTS THE PHYSIOLOGICAL OR PATHOLOGICAL MOLECULAR EVENTS OF THE PROCESSES IN THE HUMAN. SO IN PRINCIPLE IT CAN DETECT SOME OF THE -- EARLIER THAN STRUCTURAL CHANGES THAT ARE CAPTURED BY THE STRUCTURE BASED METHODS. SO FOR THE PET TO WORK WHAT YOU REALLY NEED IS A MONIKER THAT IS LABELED WITH A -- -- AND YOU CAN USE THIS TO FOLLOW HOW THOSE MOLECULES ARE DISTRIBUTED AND SPACED IN THE BODIES SO THIS IS USED IN THE CLINIC FOR THE TUMOR DIAGNOSES. NOT IN THE NORMAL CELLS. AS TIME GOES BY YOU CAN VIEW THE SIGNAL THAT IS SPECIFIC FOR THE TUMOR AS THE BACKGROUND CLEARS OUT. SO WITH THAT CONCEPT THE QUESTION IS CAN WE USE THE SIMILAR CONCEPTS AND APPLY TO THE BACTERIAL INFECTION FIELD. WE NEED A TRACER THAT IS SPECIFIC FOR THE BACTERIAL INFECTIONS AND THEN WE CAN USE THIS TECHNOLOGY TO TELL US WHERE THE INFECTION IS. IT'S QUANTITATIVE. SO BASED ON ALL OF THE WORK THE FIELD HAS BEEN DONE -- WE COME UP WITH SOME OF IDEAS ABOUT WHAT TYPE OF TRACERS WE'RE LOOKING FOR IN THIS PROCESS. WHAT WE WANT IS A -- THAT CAN DIFFERENTIATE BETWEEN INFECTION -- -- INFLAMMATION AND CANCER. WHAT WE WANT TO KNOW WHAT TYPE OF BACTERIA THAT CAUSES THIS INFECTION THAT HELPS US TAYLOR THE TREATMENT REGIMEN MORE SPECIFICALLY FOR THESE PATIENTS AND WE HAVE ALSO WANT THIS -- CAPABLE OF QUANTIFYING AND THE EFFICACY AND LAST BUT NOT LEAST WE WANT -- THIS COMPOUND IS EASY TO PREPARE AND CHEAP FOR EASY TRANSLATION TO THE CLINIC. AND WITH THAT AND ALSO THE FIELD ALSO COME UP WITH NICE IDEAS THAT CAN WE UTILIZE THESE MONIKERS THAT ARE USED BY THE PATH BEGINS. -- * PUT SOME LABELS ON THEM WITHOUT MODIFYING THEM TOO MUCH AND WITH GOOD -- FARM OH KINETIC -- WE CAN *. [ INDISCERNIBLE ] ONE OF THE PATHWAYS THAT WE'RE INTERESTED IN -- IS FOLATE PATHWAY. AND WHILE THE SUBSTRATE TO MAKE THIS IS CALLED PAB PUBLIC -- AND THE INTERESTING THING IS THERE IS -- WELL-KNOWN DRUG THE FIRST COMPOUND PASSED -- -- PABA THIS COMPOUND IS USED TO TREAT TB INFECTIONS AND STUDIES HAVE SHOWN THAT IT IS NOT A INHIBITOR. IT ACTS AS AN ALTERNATIVE SUBSTRATE. AND THEN FOLATE ANALOGUES CANNOT BE UTILIZED BY THE TB PATHOGENESIS. SO THESE ACTUALLY -- PLUS SOME OF THE EARLY DISCOVERIES -- WHERE A LOT OF SCREENING OF -- TO SWING OUT SOME OF THE PROMISE MONIKERS ONE IS -- TREATING -- LABEL FOR PABA AND SHOWS PROMISING PROPERTIES IN TERMS OF ACCUMULATING BOTH -- WHILE MANY MORE ACCUMULATION FOR THE MEMORY CELLS. SO FOR THOSE CELLULAR BEHAVIOR PROMPTED US TO DEVELOP -- OF PABA TO TEST THE EFFICACY OF THIS COME BOUND. -- COMPOUND. SO WE HAVE THE COLLABORATION OF THE -- LABOR RADIO LABELED THIS WITH FLORA 18. THAT IS SOMETHING OF THE RADIO SENSORY AND ADDITIONAL TO THAT WE ALSO DEVELOPED A BIOCHEMICAL ACIDS TO ANSWER THE QUESTION AFTER THE FLOOR AFTER 19 -- -- IT HAS A SIMILAR -- VALUE IN TERMS OF COMPARED TO THE NATURAL SUBSTRATE. WHICH PROVIDES EXTREMELY VALID BIOCHEMICAL EVIDENCE FOR US TO MOVE INTO IN VIVO. SO WE PREPARED -- WE HAVE VALIDATED -- [ INDISCERNIBLE ] WHICH IS USED IN THE FIELD TO TESTING AS A FIRST STAGE OF TRACING THE TRACING EFFICACY. AND THEN WE MONITOR THE DISTRIBUTION AND LOCATION OF THE TRACERS IN THESE MODELS. AS YOU CAN SEE FROM THE TIME -- AT TWO HOURS THERE IS ALREADY A VERY NICE CONTRAST BUILDING BETWEEN THE INFECTION VERSUS INFLAMMATION AND THE IMAGES ALSO SHOW US A VERY NICE -- IN THE AFFECTED SIDE. [ INDISCERNIBLE ] INFLAMED SIDE. SO THAT IS VERY NICE BUT THE RATIO OF THIS COMPOUND -- BETWEEN INFECTED VERSUS INFLAMED WERE HIGH HOWEVER THE ABSOLUTE UP TAKE OF THIS COMPOUND IN THE INFECTED SIDE IS NOT AS GOOD AS WE EXPECT. AS WE KNOW THE ABSOLUTE UP TAKE INSIDE THE INFECTION OF INTEREST IS DUE TO MANY PARAMETERS. SOME ARE DUE TO THE BACTERIA ITSELF AND DIFFERENT METABOLIC STATE OF THE BACTERIA BUT THE OTHER IS THE FARM OH KINETIC COMPOUND. * SO WE'RE FOCUSING ON IMPROVING THE STRUCTURES OF THE COMPOUND. AND WE PROPOSE THAT -- THOSE TWO FUNCTION GROUPS NEED TO KNOW ABSOLUTE UP TAKE. -- [ INDISCERNIBLE ] AND SO AS WE KNOW THIS GROUP IS ESSENTIAL FOR THE DSPS REACTION TO HAPPEN IN COLLABORATION. TO TACKLE THESE QUESTIONS BASED ON THE LITERATURE THAT HAS SHOWN THAT -- -- [ INDISCERNIBLE ] AND WE'RE THINKING CAN WE MODIFY OR CAN WE MODIFY THE ACID GROUP -- AND USING THE PROJECT -- WITH REPLACING WITH THE NATURAL GROUP. ONCE THIS COMPOUND GOES TO THE BACTERIA AND THE -- [ INDISCERNIBLE ] TO THE ACID AND -- [ INDISCERNIBLE ] EFFICIENTLY REDUCE TO RELEASE THE INTACT -- TRACER AND THIS TRACER WILL BE INCORPORATED INTO THE FOLATE. THIS IS OUR HYPOTHESIS AND TO TEST THEM FIRST WE ESTABLISHED -- NATURAL LAKE THAT'S REDUCING THIS COMPOUND TO THE GROUP AND IT TURNS OUT THIS COMPOUND IS A SUB STRAIGHT FOR THE MIGHT THROW -- -- WE FIRST TOOK A SIMPLE STEP OF HAVING THIS -- PRECURSORS AND THEN WE DO REACTIONS AND THIS WORKED AND THIS PROVIDES REASONABLE AMOUNT FOR OUR PRELIMINARY STUDIES AND WE HAVE PURIFIED AND CONFIRMED THAT WITH THE CO CO-INJECTIONS AND ONCE THE REACTION BECAUSE OF THE STERILE HINDRANCE WE DID NOT GET ENOUGH RADIO CHEMICAL YIELD AND SO TO PARTICIPATE -- IN THE MEANWHILE WE ALSO ESTABLISHED THREE STEP RADIO SYNTHESIS WHICH IS ADAPTED FROM OUR FIRST GENERATION PABA. IT IMPROVED DRAMATICALLY AROUND 90%. SO WITH THIS ESTABLISHED AND WE TOOK THIS COMPOUND -- Z SO THE SAME MODELS AS WE USED OFTEN AND AS YOU CAN SEE -- AFTER THREE HOURS VERY NICE SIGNALS ACCUMULATED IN THE AFFECTED SITE WHERE THERE IS MINIMAL SIGNAL IN THE -- SIDE. THERE IS AROUND LIKE 15 FOLD UP TAKE IN THE INFECTED SIDE VERSUS THE INFLAMED AND THE CHANGES ASSOCIATED WITH THE TRACE UP TAKE AND ALSO -- [ INDISCERNIBLE ] CONFIRMED OUR STUDIES. AND THEN WE VALIDATED THESE MODELS TO SHOW THAT WE DID INDUCE ENOUGH INFLAMMATION. AND THEN WE FURTHER VALIDATED OUR RESULTS UTILIZING LUMINESCENCE OF MATERIAL. BETWEEN THE PET SIGNAL AND ALSO THE -- SIGNAL AND THIS IS WHAT WE OBSERVED. AND THEN BECAUSE -- WE ALSO WANT TO KNOW HOW THIS COMPOUND METABOLIZED BECAUSE THESE NATURAL GROUPS SHOULD BE STABLE IN THE PLASMA BEFORE IT METABOLIZED -- ALTHOUGH WE KNOW THERE IS SOME -- IN THE LIVER. SO WE PREPARED ALL OF THE CODE OF STANDARD FOR THE POSSIBLE METABOLIZED. SO WE SHOWED THAT ACTUALLY -- TELLS US THAT IT QUICKLY HIDELIZED IT TO THE ACID. AND THIS FURTHER CONFIRMED THAT OUR IN VIVO STUDIES SHOWS US THAT THE NIGHT ROW IS INTACT -- TO THE ACID. SO THIS LED TO OUR SECOND HYPOTHESIS THAT MAYBE THE ACTIVE TRACER IN THE BODY IS THE NATURAL ACID. [ INDISCERNIBLE ] WE ALSO USED OUR PREVIOUS -- TO FURTHER CONFIRM AND -- [ INDISCERNIBLE ] MODELS OF UTILIZING THE LUMINESCENCE -- WE DO SEE THE -- BETWEEN INFECTED VERSUS INFLAMED. AND ALSO AS YOU CAN SEE -- SHOWS THAT THERE IS AN INCREASING RATIO OF INFECTED VERSUS INFLAMED. AND LAST BUT NOT LEAST WE ARE REALLY CURIOUS OF ASSESSING WHETHER THE FOLATE IS THE MECHANISM OF MITIGATING THOSE COMPOUNDS SO TO TEST THAT WE ACTUALLY DEVELOPED A -- SIMILAR STUDY WHERE WE KNOW THAT THE SMX WHICH IS THE INHIBITOR AND THE PABA IS A SUB STRAIGHT. SO WE CAN USE THIS TO ASSESS ANALOGUES OF ABLE TO BE A FOLATE PATHWAY. AS YOU CAN SEE THOSE ARE MIC CURVES SO AS YOU ADD MORE AND MORE PABA. THE MIC SHIFTED TO THE RIGHT WHICH INDICATES PABA CAN RESCUE THE BACTERIA AND THIS ALSO HAPPENS FOR THE F PABA. SO WE STRONGLY SUGGEST THAT F PABA IS ABLE TO BE INCORPORATED INTO THE FOLATE AND MAKE THE BACTERIA SURVIVE IN THE PRESENCE OF SMX AND THIS ALSO HAPPENS FOR OUR NATURAL ACID COMPOUNDS AS A RIGHT SHIFT OF THOSE RESPONSIBLE CURVE. SO WITH THAT I WOULD LIKE TO THANK ALL OF THE MEMBERS IN THE LAB WHO CONTRIBUTED A LOT OF WORK IN TERMS OF BIOCHEMICAL CELLULAR AND IN VIVO AND OTHER WORK THAT ARE HAS BEEN DONE AND ALSO HAVE VERY NICE COLLABORATORS ACROSS THE STUDY GROUP AND ALSO THE JOHNS HOPKINS. AND I WOULD LIKE TO THANK ALL OF THE NIH FUNDING AGENCIES TO SUPPORT OUR IMAGING WORK TO TEST OUR EFFORTS OF DEVELOPING BACTERIAL TRACERS TO THIS FIELD W THAT THANK YOU SO MUCH. I WOULD LIKE TO ANSWER ANY QUESTIONS THAT YOU MAY HAVE. THANK YOU. >> THANK YOU DR. LI. THAT WAS AN INTERESTING TALK. I ENJOYED IT VERY MUCH. AT THIS TIME WE CAN START THE DISCUSSION SESSION SO I WOULD REQUEST ALL OF THE SPEAKERS TO TURN ON THE CAMERAS PLEASE. >> THANK YOU ALL VERY MUCH. SO I WOULD LIKE TO BEGIN WITH I THINK THE FIRST QUESTION IS FOR DR. WILSON. HOW WOULD BACTERIAL INFECTION DETECTION REFLECT METABOLIC INFECTION RELATED TO DISEASE AGGRESSIVENESS. >> SO, THAT IS A REALLY GOOD QUESTION. WE THINK ABOUT THIS -- WE DO A FAIR AMOUNT OF THINKING ABOUT CANCER AND THE IMAGING OF CANCER IN MY LAB AND THE REAL TRICK IS TO IDENTIFY BAD ACTORS WITH RESPECT TO HOW CANCER PROGRESSES AND WHETHER OR NOT IT NEEDS TO BE TAKEN OUT. THAT QUESTION IS PARTIALLY ANSWERED IN A COUPLE OF WAYS. BY THIS IDEA OF BEING ABLE TO METABOLICALLILY READ OUT THE EFFECTIVE THERAPY. I THINK THAT NO MATTER WHAT IMAGING TECHNIQUE THAT WE USE WE REALLY ARE STUCK WITH TISSUE IN A LOT OF WAYS. ONCE YOU'VE IDENTIFIED AN INFECTION YOU'RE STUCK WITH DOING A BIOPSY. GETTING THE CELLS AND CULTURING THEM AND LOOKING AT THEIR SENSITIVITY TO DIFFERENT ANTIBIOTICS. ALSO POTENTIALLY DRESSED BY HOW DO BACTERIA BEHAVE IN BIOFILMS AND WE HAVE NOT WORKED IN THOSE MODELS YET BUT SOME GOOD PRECLINICAL MODELS ARE KIND OF LIKE AN AREA OF EXPANSION IN THIS FIELD. WE'RE NOT SPECIFICALLY LOOKING AT SOMETHING DIRECTLY RELATED TO BACTERIAL PATHOGENESIS. WHAT IS A SENSOR THAT WILL TELL YOU THIS BACTERIA IS -- THIS PARTICULAR STRAIN IS GOING TO BE AGGRESSIVE. WE'RE NOT LOOKING AT THAT RIGHT NOW BUT IT'S ANSWERED IN A LOT OF THE OTHER WAYS. CAN YOU DETECT IT AND DO THEY RESPOND TO ANTIBIOTICS WOULD BE SOMETHING THAT WE'VE DRESSED. -- * AND I SUPPOSE LOCALIZATION OF THE BACTERIAS IMPORTANT. >> YEAH. *. >> AND THEIR RESPONSE TO ANTIBIOTICS IS LIKE THE KEY. A LOT OF THESE PUBLICATIONS AND A LOT OF CLINICAL WORK PRESENTED BY SANJAY HAVE ADDRESSED THAT DIRECTLY AND THAT IS THE KEY OUTCOME OF WHETHER OR NOT THERE ARE BAD ACTORS AND WHETHER OR NOT THEY ARE GOING TO RESPOND TO THERAPY. >> RIGHT. THE NEXT QUESTION IS ALSO FOR YOU. THE QUESTION -- OF ORTHOPEDIC INFECTION POST HIP OR KNEE REPLACEMENT IS A MAJOR CURRENT CLINICAL ISSUE. DO YOU HAVE ANY PROGRESS HERE AND SORRY IF THIS WAS ADDRESSED BEFORE. >> YEAH. I THINK SANJAY PERSONALLY ADDRESSED THAT. GREAT QUESTION AND AS FAR AS ACCESS TO SPECIMENS AND CULTURING THEM THE ORTHOPEDIC HARDWARE REMOVAL AND CULTURING IS GREAT FROM A GOLD STANDARD PERSPECTIVE TO IDENTIFYING THIS. A LOT OF THE INVESTIGATORS THAT I'VE TALKED TO -- THEY DON'T WORK SUPER CLOSELY WITH PEOPLE IN PRECLINICAL MODELS OF BIOFILM. THE IN VIVO MODELS ARE EXCELLENT BUT THERE IS ROOM FOR OTHER SCREENING TO WORK IN THIS SPACE SO HOPEFULLY IN THE FUTURE. >> SO THIS IS A QUESTION TO ACTUALLY ALL OF THE SPEAKERS. WHAT ARE SOME OF THE CHALLENGES YOU FACE WHEN USING YOUR -- BIOFILM IMAGING? >> I CAN TAKE THAT QUESTION. I THINK THIS DISCUSSION AND THE OTHER QUESTIONS THAT WERE BEING ASKED ARE VERY IMPORTANT ONES AND I WOULD LIKE TO ACKNOWLEDGE AND I HOPE MY CO-SPEAKERS AGREE THAT METABOLIC IMAGING IS ONE OF THE LIMITATIONS IS THAT YOU NEED METABOLISM BY BACTERIA AND DEPENDING ON THE MICRO ENVIRONMENT WITHIN THE BODY I GUESS AT THAT POINT IN TIME INCLUDING BIOFILMS YOU MAY OR MAY NOT SEE METABOLISM FOR A SPECIFIC PATHWAY. AND I SAY THIS BECAUSE WHEN WE'RE TREATING INFECTIONS ANTIBIOTICS ARE ALSO TARGETING SPECIFIC METABOLIC PATHWAYS IN BACTERIA SOME WORK REALLY WELL FOR KILLING BACTERIA IN BIOFILMS AND SOME DON'T. SO DEPENDING ON THE MECHANISM OF UPTICK OF THE TRACER A TRACER MAY OR MAY NOT WORK VERY WELL IN SLOWLY DIVIDED BACTERIA. AND I'M TALKING MOSTLY ABOUT METABOLIC BASED TRACERS. THERE ARE OTHER METHODS OF BEING ABLE TO IMAGE AND WE HEARD ABOUT ANTIBODY IMAGING ETC. ETC. THOSE ARE INDEPENDENT OF IT BUT THEY HAVE OTHER LIMITATIONS. THEY MAY NOT BE ABLE TO DETECT LIVE BACTERIA. SO I THINK A LOT OF PEOPLE ARE FOCUSING ON BIOFILMS BUT I WANT TO SAY A FEW THINGS ABOUT THAT IN SPECIFICALLY TO ORTHOPEDIC INFECTIONS. THIS IS MY OPINION THAT THE METABOLIC TRACERS ARE GOING TO BE GREAT FOR DETECTING ACUTE INFECTIONS. WHEN YOU START GOING TO VERY FEW NUMBERS OF BACTERIA THEN YOU WOULD NEED CERTAIN MOLECULES THAT ARE TAKEN UP BY THESE SLOWLY DIVIDED BACTERIA AND ARE SUPER SENSITIVE. AND I WOULD LOVE TO HEAR WHAT OTHER PEOPLE THINK. THE ONLY TRACER THAT HAS BEEN TESTED AND SHOWN THAT IT IS TAKEN UP IS -- BENZONE -- ACID. THE POINT I'M TRYING TO MAKE IS WE SHOULD BE CAUTIOUS ABOUT THINKING ABOUT THE -- WHAT THE METABOLIC TRACERS CAN AND CANNOT DO AND THEIR LIMITATION AND I REMAIN OPTIMISTIC AND PEOPLE WILL FIND PATHWAYS TO BE USEFUL. >> I THINK THAT IS A REALLY GOOD POINT SANJAY. SO THE QUESTION WAS SLOWLY DIVIDING BACTERIA IS CAN WE DO BETTER. THERE IS A LOT OF ADVANTAGES TO METABOLIC TURN OVER ACCUMULATING TRACER IN THE CONTEXT OF CATALYTIC VERSUS DIRECT -- WE HAVE A SENSITIVE ADVANTAGE SO I'M NOT SURE HOW WE CAN GET THAT BETTER FOR EVALUATING BACTERIA IN BIOFILMS. THERE IS POTENTIAL ARGUMENTS THAT I'VE READ FOR TRYING TO IMAGE THE UNIQUE COMPONENTS. -- >> THANKS DR. WILSON AND JAIN. THIS NEXT QUESTION IS FOR DR. YONG LI. -- DO YOU SEE ANY PARA F18 FLOOR NATION AND CAN YOU SEPARATE THEM BY HBLC? >> YES. THIS IS A GOOD QUESTION. THIS IS EXACTLY WHAT WE OBSERVED WITH THIS OUR FIRST -- METHOD OF UTILIZING THIS. AS I SAID BEFORE THE LOW RATE OF CHEMICAL YIELD -- LARGELY DUE TO -- WHAT WE OBSERVED FOR INCORPORATING FLORA 18. 8 HOWEVER LUCKILY WE CAN SUCCESSFULLY SEPARATE THEM AS YOU CAN SEE. THEY ARE CLOSE TO EACH BUT WE SACRIFICE A LITTLE BIT TO GET A VERY PURE TRACE FOR OUR USE. AND ALSO BECAUSE OF THIS THERE IS -- ACTUALLY WE ESTABLISHED A SECOND -- BY UTILIZING A DIFFERENDIFFER-- DIFFERENT PRECURSOR. >> AND THERE IS -- THANK YOU. THERE IS ONE MORE FOLLOW-UP QUESTION. WHAT ACCOUNTS FOR THE DIFFERENTIAL UP TAKE IN DIFFERENT STRAINS OF STAPH -- ORIONIS. >> DID WE JUST LOSE HIM? I SUPPOSE SO. AND. >> HE IS CONNECTING AGAIN. >> I'M SORRY. >> WERE YOU ABLE TO HEAR THE QUESTION. >> YES. YES. I THINK THE UP TAKE REALLY IF YOU THINK ABOUT FROM THE BIOCHEMICAL PERSPECTIVE WE HAVE A DIFFERENT STRAIN. A DIFFERENT STRAIN -- BECAUSE -- IS MEDIATED BY THE DHPS. MAYBE THEY ARE -- HOW EFFICIENT ONCE THE COMPOUNDS IS INSIDE THE CELLS AND PLUS IN OUR COMPOUND HAVE ADDITIONAL LAYER OF MECHANISM UTILIZING THE NATURAL LAKLACTATES. SO. [ INDISCERNIBLE ] DEFINITELY YOU CAN GUARANTEE THERE IS SOME -- UP TAKE -- [ INDISCERNIBLE ] OF THE COMPOUND SO THIS IS MY CURRENT THOUGHT. THANK YOU. WE HAVE TIME FOR ONE FINAL QUESTION WHICH IS ACTUALLY TO DR. SUN. DID YOU EVER LOOK FOR -- GLUTAMINE TRACER UP TAKE IN FUNCTION GAL PATHOGENESIS BY ANY CHANCE BECAUSE THAT WOULD BE AN IMPORTANT DIFFERENTIAL DIAGNOSIS? >> WELL ACTUALLY THAT IS A VERY GOOD QUESTION AND THAT IS ONE -- OF WORK AND YES WE DO. >> AND DO YOU SEE -- IS THERE ANY UP TAKE? >> YES. WE DO SEE A LIMITED UP TAKE IN FUNGAL INFECTION. >> HAVE YOU LOOKED AT -- >> -- YES. >> ALL RIGHT. UNFORTUNATELY THIS IS ALL THE TIME WE HAVE FOR QUESTIONS RIGHT NOW AND MY APOLOGIES FOR ALL OF THE ATTENDEES WHOSE QUESTIONS WE DID NOT GET TO. WE WILL MAKE AN EFFORT TO FORWARD THEM TO THE SPEAKERS AND HAVE YOUR ANSWERS TO YOU BY E-MAIL. SO JUST ON A QUICK FINAL NOTE I WOULD AGAIN LIKE TO THANK ALL OF THE SPEAKERS FOR TAKING THE TIME OUT AND DISCUSSING THEIR RESEARCH AND FOR THIS VERY ENGAGING DISCUSSION SESSIONAL WELL. A BIG THANK YOU TO ALL OF THOSE LISTENING IN. I HOPE YOU ENJOYED THIS SESSION. WE WILL BE TAKING A SHORT LUNCH BREAK AND RECONVENE AT 12:30. AND THE NEXT SESSION WILL BE MODERATED BY DR. JAIN. THERE HAVE BEEN MANY WHO HAVE ARGUED THAT INFECTION IMAGING IS THE HOLY GRAIL FOR NUCLEAR MEDICINE. AND ALTHOUGH IT'S BEEN A WHILE SINCE THIS ARTICLE WAS PUBLISHED, I QUOTE THE WORK OF PROFESSOR STEVEN LARSEN FROM SLOAN-KETTERING, LABELED CANCER INFLAMMATION. THERE IS NOTHING WRONG WITH CHOOSING EITHER OF THEM OR BOTH FOR YOUR CAREER. IT'S A LOT WE NEED TO DO BECAUSE HERE IS WHERE WE ARE CLINICALLY. DIAGNOSING INFECTION IS OF COURSE TECHNIQUES, CLINICAL DESIGN, BLOCK TESTS AND OTHER RADIOLOGICAL STUDIES AND TISSUE CONFIRMATION, ALSO NUCLEAR MEDICINE TECHNIQUES THAT CAN BE NON SPECIFIC SUCH AS BRAIN SCANS, OR SPECIFIC. IN THE CATEGORY OF SPECIFIC AVAILABLE TECHNIQUES WE MENTIONED -- WHITE BLOOD CELLS ARE MOST COMMONLY USED IN SUSPECTED INFECTION. ANTIBODY IMAGING AND WE SEE APPROVAL BY THE CMS FOR REIMBURSEMENT AGAIN COMING BACK INTO THE ARENA. WHILE THE CLINICAL -- CAN BE USED FOR EVEN IF OF UNKNOWN ORIGIN, WE'LL COME BACK TO THAT, NON SPECIFIC IS RELATED TO INFLAMMATION. HISTORICALLY IT HAS BEEN USED FOR AIDS, SPECIFICALLY FOR PCP AND NAI NOTORIOUSLY NEGATIVE IN CAPOSIES SARCOMA AND THE IT DOES CLEAR THROUGH THE BOWEL. SO INFECTIONS 13-40%, DEPENDING ON WHAT LITERATURE YOU READ. AUTOIMMUNE, COLLAGEN VASCULAR DECEASE UP TO 54%. THIS IS WHERE SIGH TRAIT HAS BEEN HISTORICALLY USEFUL AND HAS BEEN LARGELY REPLACED WITH LOWER RADIATION SCORE AND HIGHER IMAGE QUALITY AND OF COURSE THE ADDITION OF CT INCREASES YOUR SPECIFICITY AS WELL AS LOCALIZATION. FOR HISTORICAL PURPOSES, AN EXAMPLE OF A PATIENT -- YOU CAN SEE THERE SHOULD BE NOTHING THERE AND OF COURSE FOR OSTEOMYELITIS WE USE THIS IN CONJUNCTION WITH A PAPER FROM THE 80s WHERE YOU CAN SEE THE ALGORITHM. IT'S USEFUL WHEN THE RADIOGRAPHS ARE NEGATIVE AND THEN THE BONE SCAN IS POSITIVE. AND MOST COMMONLY ASSOCIATED WITH INFECTION WHEN THE UPTAKE IS IN CONGRANT AS COMPARED TO THE MPP THIS HAS BEEN THE MAINSTREAM OF IMAGING BONE INFECTION FOR A WHILE. WHITE BLOOD CELL COUNTS HAS VALUES IT'S NOT CLEARED BY THE KIDNEYS OR IN THE GUT SO IF WE LOOK FOR UNKNOWN ORIGIN, THIS IS THE BETTER CHOICE. IN NORMAL BODY SOLUTION INCLUDE THE SPLEEN AS MOST INTENSE AND BONE MARROW. IF WE IMAGE LATER, THE STANDARD PROCEDURE, 24 HOURS LOW ACTIVITY. IF WE IMAGE EARLIER, THERE IS MORE IN THE LUNGS. THESE ARE SOME EXAMPLES. IN THIS CASE THERE IS FOCAL UPTAKE IN THE MID ABDOMEN ON THE PROJECTION METER ON THE LEFT AND THEN ON THE AXIAL AND CORONAL IMAGES, THIS CORRELATES TO AN ABSCESS IN THE PARTINIUM. WE NOW HAVE SPECIFIC HIGH QUALITY CT, VERY GOOD IMAGE QUALITY IN THE AXIAL AS WELL AS CORONAL VIEW DEMONSTRATING BILATERAL INFECTION FOCAL INTENSE. SO WHITE BLOOD CELLS CAN BE USEFUL IN A VARIETY OF ETIOLOGIES AND CAN SERVE WELL HIGH QUALITY INFECTIOUS DISEASE. THERE IS ALSO WORK THAT HAS BEEN DONE WITH FDG LABELED WHITE BLOOD CELLS. ON THE RIGHT IS A NORMAL LABEL AFTER INJECTION INTO HUMAN. HERE FOR ILLUSTRATION PURPOSES ON THE LEFT, WE HAVE FOCAL UPTAKE IN THE LUNGS AS WELL AS IN THE STERN UM. THIS WAS NOT BLOOD CANCER. THIS IS A PATIENT WITH -- AFTER SIX MONTHS OF TREATMENT AGAIN THE IT WORKS WELL IN THIS INSTANCE AND DOCUMENTS THE EXTENT OF INFECTION AND ALSO RESPONSE TO TREATMENT. MY COLLEAGUES AND I LOOKED AT OUR POSITIVITY RATES FOR WHITE BLOOD CELLS ORDERED FOR OCCULT INFECTION BECAUSE WE WANTED TO SEE WHERE TO PLACE THE FIRST-IN-HUMAN STUDIES IN TRIALS. FIRST LET'S LOOK AT WHAT WE FOUND OVER THE SPAN OF LESS THAN A YEAR. THERE WERE 50 PATIENTS WHO HAD WHITE BLOOD CELL SCANS WHO WERE ANEMIC AND WE DID THE TYPICAL PROCEDURE AND WE USED PLAIN IMAGING AS WELL AS SPECIFIC LOCALIZATION OF THE WHITE BLOOD CELLS. OVERALL, THE POSITIVITY WAS 42%. SO 21 OF THE 50 PATIENTS HAD POSITIVE WHITE BLOOD CELL SCANS AND INTERESTINGLY, AND SOMEHOW UNEXPECTEDLY, IN PATIENTS RECEIVING BROAD SPECTRUM ANTIBIOTICS, IT WAS HIGHER AT 53.8% VERSUS THOSE NOT ON ANTIBIOTICS WHO HAD POSITIVITY RATE OF 27.3%. THE HIGHEST POSACTIVITY RATE OVERALL 9 OF THE 12 PATIENTS WERE PATIENTS WITH KNOWN SEPSIS OR BACTERIUM. HOWEVER, THE POSITIVITY RATES WERE MUCH LOWER WHEN WE LOOKED AT SUSPECTED AUTOGRAPH INFECTION AND ENDOCARDITIS, 19%, AND: [ READING ] WHEN WE ADD SPECIFICITY, LOCALIZATION IN CASES WHERE PLAIN IMAGING WAS UNABLE TO PROVIDE. SO IF YOU HAD ACCESS TO HIGH QUALITY, DO NOT HESITATE TO USE IT. IT WILL INCREASE SPECIFICITY. WITH THIS RESULT WE WENT BACK TO OUR COLLEAGUES IN INFECTIOUS DISEASE AND THIS IS FROM OUR COLLEAGUE DR. DORA HAUL. WE SHOWED HER THE RESULTS AND THESE ARE HER COMMENTS. THERE ARE TWO SCENARIOS: [ READING ] THIS IS ONE INDICATION WHERE THE POSACTIVITY RATE WAS LOWER AND THEN THE SECOND SCENARIO IS: [ READING ] WITH THIS IN MIND, OUR RESULTS MAKE SENSE. THOSE ANTIBIOTICS EXPLAINS THE HIGHER POSITIVITY RATE OF 58% AND ALSO THE HIGHEST POSITIVITY RATE OF 75% OF PATIENTS WITH SEPSIS OR BACTERIUM. SO NOW LET'S TALK ABOUT -- THIS WAS DEVELOPED BY ONE OF MY COLLEAGUES WORKING IN SAM'S LAB AND THIS IS HER AND SAM AND ONE OF THE GROUP PICTURES TO DOCUMENT HOW LARGE THE LAB THAT SAM WAS IN A LOT OF INTERESTING WORK COMING FROM THIS LAB. SO WHAT IS THE TARGET HERE? IT IS THE PULTE TRANSPORT SYSTEM. THIS TRANSPORTER TAKES MALLITOSE AS WELL AS OTHER COMPOUNDS INSIDE THE BACTERIAL CELL AND THEN UNDER ACTIVITY IT TURNED TO GLUCOSE AND GLYCOLYSIS AND FROM HERE ON WE KNOW WHAT HAPPENS. THERE WERE OTHER PROBES DEVELOPED TARGETING THIS TRANSPORTER SYSTEM AND THESE ARE EXAMPLES FROM THE PRE-CLINICAL STUDIES THAT USES THIS COMPOUND. SO HERE ARE SOME THAT WERE HAVE BEEN TRIED. -- [ READING ] AND UNDER THE SUPERVISE OF A FANTASTIC CHEMIST, THE TRIALS WERE DEVELOPED. THIS WAS FOUND TO BE THE MOST PROMISING OTHER TESTED COMPOUNDS AND IS TAKEN FORWARD FOR CLINICAL TRANSMISSION. THE UPTAKE IS SPECIFIC TO BACTERIA. AS YOU CAN SEE IN THIS GRAPHS AND YOU CAN SEE BROAD SPECTRUM OF BACTERIA WHERE THIS IS POSITIVE. SO HOW DOES IT COMPARE? HERE ARE FOR COMPARISON ON THE LEFT, AND I THINK YOU CAN APPRECIATE THERE IS ACTUALLY UPTAKE IN THE IMPLANTED AREA OF INFECTION. THERE IS NOT CLEARLY IDENTIFIED. SO VERY PROMISING PRE-CLINICAL RESULTS AND WE ARE LOOKING FORWARD TO MORE WORK. THIS IS AGAIN EXCELLENT PHARMACOKINETIC. YOU CAN APPRECIATE THE COMPARISON WITH THE WHITE BAR VERSUS THE BLACK BAR WITH SIMILAR CLEARANCE THROUGH THE KIDNEYS BUT MUCH LOWER CLEARANCE THROUGH THE LIVER AND OVERALL DECREASE BACKGROUND IN NORMAL TISSUE. AND TO ILLUSTRATE IT CAN DISTINGUISH FROM HEAT AND ACTIVATED BACTERIA AS POINTED BY THE ARROWS ON THE LEFT, BACTERIA STRONG SIGNAL AND ON THE RIGHT ALSO WITH LOWER CONCENTRATION AND ALSO FOCAL. OVER THE PAST YEAR PLUS, MY COLLEAGUES IN PRE-CLINICAL BIOCHEMISTRY HAVE WORKED ON IMPROVING THE RADIOLABELING, PARTICULARLY YOU CAN APPRECIATE ON THESE GRAPHS THE NEW LABEL VERSUS THE OLD LABELING THAT THERE IS IMPROVED QUALITY IN THE RADIO CHEMISTRY, RADIOLABEL PROCESS. THERE IS ALSO SERUMS ABILITY THAT YOU CAN APPRECIATE PARTICULARLY IN HUMAN SERUM AT INCUBATION. THESE ARE VERY ENCOURAGING RESULTS FROM THE PRE-CLINICAL ARENA TO CLINICAL USE. SO WHERE WE ARE NOW AND WHAT ARE THE FUTURE DIRECTIONS? WE DID A TOX STUDY TO ENSURE THE COMPOUND WAS SAFE IN HUMANS. MY COLLEAGUES IN THE RADIO CHEMISTRY ARE NOW WORKING ON OPTIMIZING SYNTHESIS AND PREPARING CM C-SECTION. WE PLANNED TO SUBMIT IND FOR APPROVAL IN THIRD QUARTER OF THIS YEAR AND IT WILL START WITH VOLUNTEERS TOEST MATE SYMMETRY. AND AFTER WE DO ALL THIS CALCULATION, WE WILL SCAN 10 PATIENTS REFERRED FOR WHITE BLOOD CELL SCANS AND THEN IT WILL TAKE 5 POSITIVE SCAN AND 5 WITH NEGATIVE TO COMPARE WITH THE PET CT. FROM THERE, ASSUMING SUCCESSFUL EXPERIENCE, WE WILL SPANNED TO OTHER INDICATIONS. SO I REALLY HOPE BY NEXT YEAR, WE WILL BE ABLE TO PRESENT CLINICAL DATA FROM THE USE OF THESE PHARMACEUTICALS TO US IS VERY, VERY EXCITING. SO I WANT TO END ACKNOWLEDGING OUR STAFF IN THE RADIO CHEMISTRY FACILITY AND WANT TO HIGHLIGHT TOM AND BEN WHO ARE GREAT. TOM HOSPITAL US TREMENDOUSLY TO DO THE TOX -- TOM HELPED US TREMENDOUSLY AND HE IS ALSO WORKING TO OPTIMIZE THE RADIO CHEMISTRY AND THE CMS AND THE FANTASTIC WORK AND CARDIOVASCULAR SPACE. WITH THAT, I THANK YOU FOR THE ATTENTION AND I LOOK FORWARD TO LEARNING MORE FROM THE OTHER PRESENTERS AND LOOK FORWARD TO BEING BACK IN THIS OFFICE. THANK YOU FOR INVITATION. >> MANY THANKS FOR THIS PRESENTATION AND I'M SORRY THAT HE'S NOT HERE IN PERSON -- OR LIVE. BUT WE ACTUALLY FINISHED A FEW MINUTES EARLY. I'M GOING TO GO AHEAD AND INVITE OUR NEXT SPEAKER, DR. MILOS FROM THE CZECH REPUBLIC. DR. PETRIK HAS DONE AMAZING WORK ON LABELED IMAGING AND HE WILL TALK ABOUT BACTERIAL AND FUNGAL IMAGING USING THIS METHOD. SO WITHOUT LONG INTRODUCTION, I'D LOVE TO HAVE DR. PETRIK TAKE OVER. SO THANK YOU AGAIN FOR JOINING US FROM THE CZECH REPUBLIC. I KNOW IT'S GETTING LATE THERE SO WE REALLY APPRECIATE YOU DOING THIS. >> CAN YOU HEAR ME AND SEE ME? >> YES. >> GOOD AFTERNOON, LADIES AND GENTLEMEN. THANK YOU FOR KIND INTRODUCTION SANJAY. IT'S A GREAT HONOR TO BE A PART OF THIS GREAT SYMPOSIUM AND I WOULD LIKE TO THANK THE ORGANIZE ORGANIZERS TO PRESENT OUR WORK-RELATED TO IMAGING BACTERIAL AND FUNGAL INFECTIONS USING LABELED SIDER FORES. THEY ARE COMPOUNDS OF DIVERSE BACKGROUNDS AND THE MAIN GROUPS ARE BASED ON MIXED TYPES. THEY ARE PRODUCED BY PLANTS, BACTERIA AND FUNKY. HAVE VERY HIGH AFFINITY TO -- TRANSPORT MICROORGANISMS AND HAVE NO FUNCTION IN -- CELLS -- MAMMALIAN CELLS. IN INFECTION, IRON PLAYS A MAJOR ROLE. IT HAS LOW BIOAVAILABILITY AND PATHOGENICITY, IT IS ESSENTIAL FOR BOTH PATHOGEN AND ITS HOST AND INFECTION IS BATTLE FOR IRON. IMPORTANT FACET OF THE INNATE IMMUNE SYSTEM IS TO LIMIT IRON AVAILABILITY TO INVADING MICROBES. [ READING ] ON THIS EXAMPLE I WOULD LIKE TO BRIEFLY EXPLAIN AND CONSIDER FOR UPTAKE OF IRON INTO THE FUNGAL -- IN THIS CASE -- IS PRODUCED AND RELEADS AS PER GILL US FUME GATE US BY SPECIFIC TRANSPORTER AND IT IS THEREFORE COMPLEX IRON COMPLEX, THEN IT IS TAKEN UP BY OTHER SPECIFIC TRANSPORTER, IN THIS CASE IN ASPERGILLUS FUME GATE US TRANSPORTER WHICH IS -- IRON EFFICIENCY. AND THEN THE IRON IS COMPLEX AND TRANSPORTED INTO THE FUNGAL -- IT IS CLEAVED AND THEN THE IRON IS USED FOR METABOLIC PROCESSES. THE PRINCIPLE EXISTS ALSO IN BACTERIA. THE IDEA OF RADIOLABELS FOR INFECTION IMAGING IS MAINLY BASED ON THE CHEMISTRY BETWEEN IRON AND GALLIUM. THEY HAVE IDENTICAL COMPLEX CHEMISTRY. IRON RADIUS AND OTHER PHYSICAL PROPERTIES, HOWEVER THEY ALSO DIFFER OF COURSE. GAL YUM -- ACID AND [ READING ] OUR MAIN GOAL IN THIS PROJECT WAS TO INVESTIGATE IF THE IRON CAN BE REPLACED BY GALLIUM 68 TO CREATE STABLE GALLIUM 68 LABELED FOR IMAGING OF MICROBIOME INFECTIONS. OUR APPROACH WITH GALLIUM 68 IS BASED ON TROJAN HORSE STRATEGY. THE UPTAKE TO THE -- IS BASICALLY THE SAME OR IDENTICAL AS EXPLAINED ON THE PREVIOUS SLIDE. THE ONLY DIFFERENCE IS THAT THE FUNGAL R. US COLLECTS GALLIUM 68 INSTEAD OF IRON. ON THE NEXT SLIDES I WOULD LIKE TO SHOW YOU OUR RESULTS WITH GALLIUM IMAGING OF FUNGAL AND BACTERIAL INFECTIONS AND I WILL START WITH FUNGAL INFECTIONS. THE PROOF-OF-CONCEPT STUDY WAS DONE WITH -- AND PRODUCED BY -- WE WERE ABLE TO RADIO LABEL -- WITH GAT YUM 68 RADIO CHEMICAL HIGHER THAN 98%. WE HAVE SHOWN HIGH UPTAKE OF -- IRON DEFICIENT CULTURES WILL BE BLOCKED WITH IRON OR SODIUM -- AND THE UPTAKE WAS INCREASING IN TIME. THESE ARE ALSO SUCCESSFUL IMAGING WITH INFECTION AND PULMONARY INFECTION MODEL AS YOU CAN SEE ON THE -- IN CONTINUING RESEARCH, WE FOCUSED ON INDIVIDUAL AND IN-VIVO CHARACTERIZATION OF GALLIUM 68 LABELED SERUM WHICH COULD BE POTENTIALLY TAKEN UP BY AS PER GILL US FUME GATE US INFECTION IMAGING. WE HAVE SHOWN THAT ESPECIALLY GALLIUM TAFC AND GALLIUM FOX E ARE PROMISING CANDIDATES SHOWING EXCELLENT PHARMACOKINETIC PROPERTIES IN HEALTHY MICE. WE HAVE STUDIED THE SPECIFICS OF BOTH COMPOUNDS IN DIFFERENT MICROBIOME CULTURES AND LUNG CANCER CELLS. WE HAVE OBSERVED HIGH INDIVIDUAL UPTAKE OF BOTH COMPOUNDS IN AS PER GILL US FUME GATE US AS YOU CAN SEE ON THE FIRST LINE OF THE TABLE AND WHILE GALLIUM TAFC SHOWED QUITE SPECIFIC UPTAKE, IT WAS TAKEN UP MORE THAN -- GALLIUM FOX E WAS TAKEN UP BY LEGAL ALL FUNGAL SPECIES UNDERSTUDY AND MOREOVER IT WAS ALSO TAKEN UP BY -- BOTH COMPOUNDS SHOWED CLEAR FOCAL UPTAKE IN INFECTED LUNG TISSUE INFECTED RATS AS YOU CAN SEE ON THE IMAGES. WE WERE ALSO ABLE TO MONITOR THE INFECTION IN ANIMALS WITH DIFFERENT LOAD OF SPORES. THE MODEL IMAGES FOLLOWED INFECTION UP TO THREE DAYS POST-INFECTION WHILE THE MODEL MODEL -- UP TO 10 DAYS AFTER INFECTION. NOW I'M COMING TO THE BACTERIAL INFECTIONS. THIS IS A LIST OF COMMON BACTERIALS INCLUDING QUITE SPECIFIC CIDER FORCE AND -- BUT ALSO SIDO FORCE RANGE OF BACTERIA. TYPICAL EXAMPLES OF SUCH ARE: [ READING ] ON THE NEXT SLIDES I WOULD LIKE TO SHOW YOU OUR RESULTS WITH ONE REPRESENTATIVE OF HIGHLY HIGHLY-SPECIFIC SIGH DERFORES. FIRST ONE HIGHLY SPECIFIC EXAMPLE IS GALLIUM LABELED -- LABEL THIS WITH GALLIUM 68 WITH VERY HIGH RADIO CHEMICAL PURITY, HIGHER THAN LIKELY, HIGH UPTAKE OF THIS COMPOUND IN CULTURES, INCREASING RATE TIME AND WE BLOCKED WITH IRON -- THE SPECIFICITY WAS TESTED IN 15 DIFFERENT MICROCULTURES SHOWING REALLY GREAT SPECIFICITY JUST FOR THE -- [ INAUDIBLE ] WE HAVE ALSO TESTED OTHERS IN THE -- AND WE COULD AGAIN SEE JUST VERY HIGH UPTAKE IN THIS STRAIN. AND THEN WE SHOWED HIGH ACCUMULATION OF GALLIUM IN -- INFECTED LUNG TISSUE SHOWING CLEARLY DIFFERENT WHEN COMPARED WITH HEALTHY ANIMALS EX-VIVO AND ALSO ON IMAGES. YOU CAN SEE THE HEALTHY IN LUNG AREA WHILE ON THE RIGHT SIDE IT IS CLEAR ACCUMULATION -- WE HAVE ALSO STUDIED SPECIFICITY OF GALLIUM IN-VIVO IN MOUSE ALTHOUGH THE LEFT LEGS OF EXPERIMENTAL ANIMALS INJECTED AND THE RIGHT LEGS -- TO CAUSE INFLAMMATION. WE HAVE INJECTED SUCH ANIMALS WITH GALLIUM IN ANIMAL A AND B AND COMPARED THE RESULTS BETWEEN RADIOPHARMACEUTICALS FOR INFECTION IMAGING SUCH AS GALLIUM SIGH TRAIT. AS YOU CAN SEE FROM THE IMAGES A&B GALLIUM SHOWED SPECIFIC INFLAMMATION IN THE LEFT LEGS, MEANING MUCH BETTER DISTRIBUTION COMPARED TO CLINICAL USE RADIOPHARMACEUTICALS. THE LAST EXAMPLE FOLLOWED WORK-RELATED TO GALLIUM 68. DFO IS USE FOR MEDICAL USE BY FDA ALREADY IN 1968 UNDER BRAND NAME -- BASED ON THIS COINCIDENCE, WE DECIDED TO STUDY IF THE GALLIUM 68 LABEL DFO WOULD WORK FOR INFECTION BECAUSE IF IT WOULD WORK, IT COULD REALLY HAVE QUITE GOOD POTENTIAL FOR EASY CLINICAL TRANSLATION. WE COULD LABEL AGAIN THE DFO GALLIUM 68 WITH CLINICAL PURITY. WE SHOWED IN VITRO IT COULD BE TAKEN OUT BY DIFFERENT STRAINS OF STAPHYLOCOCAUS AND STREPTOCOCCUS. IT CAN BE BROUGHT TO UPTAKE WITH IRON DFO. IN HEALTHY MINDS, IT SHOWED EXCELLENT PHARMACOKINETICS BY KIDNEYS. NOTHING IN THE ORGAN. AS YOU CAN SEE EX-VIVO IN DISTRIBUTION DATA. IN-VIVO UPTAKE WAS STUDIED IN THIS MOUSE MODEL. THE LEFT BEING LEGS OF EXPERIMENTAL ANIMALS INJECTED -- [ INAUDIBLE ] THE RIGHT LEGS WERE INJECTED -- GALLIUM DFO SHOWED CLEAR SPECIFIC UPTAKE INES PIN OWESSA AND INFECTED MUSCLES. YOU CAN SEE ON THE IMAGES. I AM HAPPY THAT I CAN FINALLY SHOW YOU SOME FIRST PATIENT DATA DATA. GALLIUM CURRENTLY UNDERGOING A CLINICAL TRIAL AND HERE ARE THE FIRST PROMISING IMAGES OF PATIENT WITH STAPHYLOCOCAUS LEFT IN INFECTION COMPARED WITH SCAN. NOW RELATED TO -- I ALSO PREPARED SIDER FORES FOR IMAGING AND ANTIFUNGAL APPLICATIONS AND BASED ON ADDITIONAL BRANCH OF OUR RESEARCH WE BELIEVE THAT SIDER FORES -- ALSO USING ANALYTICAL TECHNIQUES SUCH AS LCMS AND ESPECIALLY THE COMBINATION WITH IMAGING WITH INTERESTING TOOL, PROBABLY. TO CONCLUDE, MANY DIFFERENT SIDER FORES CAN BE LABELED GALLIUM 68 FOR PET IMAGING. [ READING ] I WOULD LIKE TO THANK ALL COLLEAGUES WHO PARTICIPATED ON THIS PROJECT. SPECIAL THANKS GOES TO MY GREAT COLLEAGUE AND COLLABORATOR PROFESSOR CLEMENS FROM MEDICAL UNIVERSITY IN AUSTRIA AND THANK YOU FOR YOUR ATTENTION. >> THANK YOU SO MUCH FOR THAT EXCELLENT TALK. I'M SO EXCITED ABOUT BOTH THE SIDER FORES TO IMAGE BUT ALSO THE FACT THAT YOU PUT THIS INTO THE CLINIC. WE REALLY LOOK FORWARD TO THE RESULTS FROM THE CLINICAL STUDIES. INTRODUCE THE LAST SPEAKER FOR THIS SESSION ALSO FROM EUROPE. SO I REALLY APPRECIATE ALL THE FOLKS WHO HAVE BEEN TALKING. IT'S CHRISTOPHER THORNTON FROM THE UNIVERSITY OF EXETER TALKING ABOUT FUNGAL INFECTIONS USING LABELED ANTIBODIES. >> THANK YOU VERY MUCH. I HOPE EVERYTHING IS WORKING AND YOU CAN SEE EVERYTHING OKAY? >> YES, WE DO. >> THANK YOU. SO THANK YOU VERY MUCH INDEED FOR THE INVITATION TO SPEAK TO YOU TODAY. SO WHAT I'M GOING TO DO IS TALK ABOUT IMAGING FROM INFECTIONS USING LABELED ANTIBODIES AND SPECIFICALLY TALKING ABOUT ONE PARTICULAR MOLD PATHOGEN OF HUMANS, ASPERGILLUS, INVASIVE FORM. SO I'M AT THE UNIVERSITY OF EXETER IN THE SOUTH WEST OF EN LAND. WE HAD JOE BIDEN FOR THE G7 DOWN THE ROAD AND ALREADY SAID HE LOVES IT SO MUCH HE DOESN'T WANT TO COME HOME. SO YOU MIGHT NOT BE GETTING YOUR PRESIDENT BACK. SO WE ARE DOWN IN THE SOUTHWEST. I'M AT THE UNIVERSITY ACADEMIC AND A MEMBER OF THE MEDICAL RESEARCH COUNCIL CENTER FOR MEDICAL MYCOLOGY LARGEST GROUPINGS OF LIKE-MINDED INDIVIDUALS LOOKING AT MEDICAL MYCOLOGY IN THE WORLD. I'M ALSO, HAVE TO DECLARE DIRECTOR OF THE UNIVERSITY OF -- FOR DIAGNOSTICS. SO ASPERGILLUS DISEASE. THERE IS A NUMBER OF SPECIES THAT CAUSE DISEASE IN HUMANS. THE MOST IMPORTANT IS ASPERGILLUS FUNG AT US THAT CAUSES 18% PLUS INFECTIONS IN HUMANS. SO IT'S THE MOST IMPORTANT OPPORTUNISTIC MOLD PATHOGEN IN IMMUNOCOMPROMISED HUMANS ESPECIALLY THOSE WITH HEME TA LOGICAL MAGLIGNANCIES AND BONE MARROW TRANSPLANT PATIENTS. SO THE DISEASE IN IS ONE OF A NUMBER OF DISEASES CAUSED BY ASPERGILLUS. SO RANGING FROM ACUTE TO CHRONIC INFECTIONS. SO INVASIVE PULMONARY PAS PER GILOWSIS -- ROUGHLY 300 CASES PER YEAR -- 300,000 CASES PER YEAR WORLDWIDE. IT RANGES FROM 30-95% IN SOME SENSES SO THE MORTALITY ASSOCIATED WITH THAT DISEASE THAT IS REALLY IMPORTANT. YOU THEN HAVE CHRONIC PULMONARY AS PER JILLOSEIS, ROUGHLY 3 MILLION CASES WORLDWIDE. PARTICULARLY IN PATIENTS WITH UNDERLYING LUNG DISEASES INCLUDING ASTHMA AND THEN WE HAVE ALLERGIC -- ADPA, ROUGHLY 4 MILLION CASES WORLDWIDE. THE PATHOGEN IS ALSO EMERGED AS A SUPER PATHOGEN CAUSING SUPER INFECTIONS IN COVID-19 PATIENTS AND WE HAVE THIS DISEASE NOW COVID-19 ASSOCIATED PULMONARY AS PER JILLOSEIS, OR CAPA AND 30% OF ICU PATIENTS COVID HAVE DEVELOPED SUPER INFECTIONS CAUSED BY ASPERGILLUS SPECIES. IT'S BEEN KNOWN FOR A WHILE IT IS ALSO INVOLVED IN THIS OTHER DISEASE INFLUENZA-ASSOCIATED PULMONARY AS PER JILLOSEIS. SO THE IMPORTANT THING IS IT'S OPPORTUNISTIC. I'M GOING TO BE CONCENTRATING ON THE BASIC FORM OF THE DISEASE ON THE RIGHT-HAND SIDE HERE. WE HAVE A BIOPSY OF THE LUNG AND YOU CAN SEE THE ELEMENTS THAT ARE GROWING THROUGH THE LUNG TISSUE AND ULTIMATELY THE PATHOGEN WILL DISSEMINATE. SO IT CAUSES WIDESPREAD DAMAGE WITHIN LUNG TISSUES. ON THE LEFT-HAND SIDE HERE WE HAVE A TYPICAL AS PER JILLOMA, A GROWTH OF FUNGAL BALL WITHIN THE LUNG AND TYPICALLY GROWS OVER A NUMBER OF YEARS I'M GOING TO BE CONGES TRAITING ON INVASIVE ASPERGILLOSIS. SO DIAGNOSIS OF THIS DISEASE IS REALLY DIFFICULT AS IS DIAGNOSIS OF ANY INVASIVE FUNGAL DISEASE IN HUMANS. YOU HAVE TO RELY ON A WHOLE LOAD OF DATA FROM DIFFERENT SOURCES TO TRY AND IDENTIFY WHETHER A SUSCEPTIBLE PATIENT HAS AN INVASIVE FUNGAL DISEASE. SO YOU'RE RELYING ON PATIENT HISTORY, WHAT IS THEIR UNDERLYING DISEASE, ARE THEY LEUKEMIC? WHICH RAISES SUSPICION OF A FUNGAL INFECTION. AND PARTICULARLY RELY ON BIOMARKERS IN INVASIVE BANKO ALPH EOLA FLUID OR CIRCULATING IN THE BLOODSTREAM. AND EACH NOW, IT IS REALLY CULTURE OF THE PATHOGEN FROM INVASIVE BIOPSY WITH ASSOCIATED NECROSIS WHICH IS THE GOLD STANDARD FOR FUNGAL DISEASE DIAGNOSIS. SO WE GOT A LOT OF IMPROVEMENTS THAT NEED TO BE MADE. SO THE NAME WE HAVE A BIT OF AN ISSUE IN THE TREATMENT ARE OFTEN FEVER DRIVEN. WE HAVE A PATIENT THAT IS AT HIGH RISK. MAY HAVE HAD A BONE MARROW TRANSPLANT. THEY ARE FEEBAL AND NON RESPONSIVE TO ANTIBIOTICS. SO THEN WE START TO SUSPECT THEY MIGHT HAVE A FUNGAL INFECTION. THE PATIENT SENT FOR CHEST CT AND THE RADIOLOGIST SAYS YES, THEY HAVE AN ABNORMALITY IN THE CHEST CT. WE HAVE NO IDEA WHAT IT IS. BUT YOU HAVE GOT A PATIENT THERE THAT IS IN A SUSCEPTIBLE PATIENT GROUP TREATMENT WITH ANTIFUNGALS AND THE TREATMENT MAY WELL HAVE STARTED BEFORE THE CHEST CT DRIVEN BY FEVER AND UNRESPONSIVE TO ANTIBIOTICS. AND THE PROBLEM IS, LEADING TO PATIENTS RECEIVING INAPPROPRIATE TREATMENT WITH COSTLY IN TOXIC ANTIFUNGAL DRUGS AND IT IS ALSO BELIEVED AS YOU MIGHT EXPECT, TO EMERGENCE OF DRUG RESISTANCE AND CLINICAL STRAINS OF ASPERGILLUS. THAT'S A PROBLEM BECAUSE WE ARE RUNNING OUT OF OPTIONS FOR TREATING INVASIVE FUNGAL DISEASES OF HUMANS. VERY FEW DRUGS AND VERY FEW TARGETS THAT TARGET THE FUNGUS SPECIFICALLY. SO IMPERATIVE THE DIAGNOSIS IS MADE AND PROGNOSIS AND RAPIDLY OVER TIME AND LEADING TO HIGH MORTALITY RATES AND THAT IS EXACTLY EXACERBATED BY POOR DIAGNOSIS. WE SET OUT SOME TIME AGO TO IMPROVE DIAGNOSE BASED ON BIOMARKERS. WE ARE TRYING TO GO TO A SITUATION WHERE WE HAVE DIAGNOSTIC-DRIVEN DRUG DELIVERY RATHER THAN PROPHYLACTIC IN MANY CASES. SO WHAT I DID WAY BACK IN 2005 IS DEVELOP MONOCLONAL ANTIBODIES SPECIFIC TO ASPERGILLUS SPECIES AND THE NICE THING ABOUT THIS MONOCLONAL ANTIBODIES, IT BINDS TO AN ANTIGEN, GLYCOPROTEIN THAT IS AN INDICATOR OF ADDED GROWTH. SO ON THE TOP PANEL HERE, WE HAVE BRIGHT FILLED IMAGES OF VARIOUS STAGES OF SPA GERM NATION AND DEVELOPMENTS FROM ASPERGILLUS. SO NUMBER 1 WE HAVE AN INERT SPORE. THESE ARE SPORES TAKEN IN DURING OUR NORMAL BREATHING DURING THE DAY. ANYTHING UP TO 2-300 SPORES PER PERSON PER DAY WITH THIS PATHOGEN ALONE. AND AFTER 12 HOURS IN THIS ENVIRONMENT, THE SPORES WILL BIND AND THEY WILL SWELL AND THEN GO TO POLAR GROWTH AND START TO PRODUCE A GERM SHOOT. THAT GERM SHOOT WILL GROW INTO A HIVER AND THEN YOU END UP WITH A MASSIVE HIGHER CAUSING INFECTION. SO IF YOU THEN PROBE THOSE SAMPLES WITH THE JF ANTIBODY CONJUGATED TO FITC AND OBSERVE THOSE TEMPERATURES UNDER FLUORESCENCE, YOU CAN SEE ONLY WHERE THE FUNG AS STARTS TO GO INTO POLARIZED GROWTH THAT WE GET PRODUCTION OF THIS GLYCOPROTEIN. AND AS THE FUNG AS DEVELOPED, YOU GET COPIOUS PRODUCTION FROM THE DEVELOPING. SO WE HAVE A REALLY NICE TARGET AND ANTIBODY ANTIGEN SYSTEM FOR DIFFERENTIATING INACTIVITY, SPORES THAT ARE NORMALLY INHALED DURING OUR BREATHING AND THOSE THAT ARE COMING IN ACTIVE AND POTENTIALLY CAUSING INFECTION. THE BOTTOM HERE WE HAVE AN INVASIVE HIGHER, PROBED WITH JF5 AND EPIFLUORESCENCE AND MASSIVE PRODUCTION OF THAT GLYCOPROTEIN. SO IT'S A REALLY GLYCOPROTEIN TARGET AND INDICATOR OF DRUG. SO WE TURNED THAT INTO A LATERAL FLOAT DEVICE AND EVERYONE IS FAMILIAR WITH THAT DEVICE NOW. TWO LINES POSITIVE, ONE LINE NEGATIVE WORKS IN EXACTLY THE SAME WAY AS COVID-19 LATERAL FLOW TEST. I'M SURE NOW EVERYONE IS NOW DOING. IN THIS CASE, WE ARE PICKING UP THE GLYCOPROTEIN USING THE JFI CONJUGATED. WHAT I DID IS INTRODUCED THE SAMPLE CONTAINING THE GLYCOPROTEIN AND WE HAVE A SAMPLE WITH THE GLYCOPROTEIN AND ONE THAT IS NEGATIVE. SO WE HAVE A SAMPLING AND CONTROL. SO THAT PARTICULAR TEST NOW MARKED AND USED AROUND THE WORLD AS A TEST FOR INVASIVE ASPERGILLOSIS USING THAT MONOCLONAL ANTIBODY. THERE IT IS: AND SO WE HAVE TAKEN DIAGNOSIS NOW TO MINUTES RATHER THAN HOURS AND DAYS OR EVEN WEEKS AND SOMETIMES EVEN MONTHS IN SOME SENSES. SO THAT'S ALL WELL AND GOOD. THAT'S A POINT OF CARE LAT ROW FLOW DEVICE. WE WANTED TO TRY TO IMPROVE USING THE SAME MONOCLONAL ANTIBODY RADIOLOGICAL DETECTION IN-SITU, IN-VIVO DETECTION IN INFECTED HUMANS. SO TYPICALLY WHAT HAPPENS ON A CT IS YOU GET SOME AB NORMALITY. SO SOME INFILTRATES OR LESION BUT IT'S COMPLETELY NON SPECIFIC. HERE WE HAVE CT OF A PULMONARY LYMPHOMA, ADEANA CARCINOMA, CRYPTOICOSEIS, AMPLEIO INVASIVE ASPERGILLOSIS AND PRIMARY PULMONARY TUBERCULOSIS AND THIS IS CAUSED BY PATHOGENS CAUSING SERIOUS PROBLEMS IN COVID-19 PARENTS AT THE MOMENT. SO 11,000 CASES OF BLACK FUNGUS INFECTION IN COVID-19 PATIENTS WHEREAS THEY NORMALLY JUST SIT 50 PATIENTS IN THE ENTIRE COUNTRY A YEAR, THEY NOW HAVE 11,000 WITH COVID-19. SO WE HAVE TO BE ABLE TO IDENTIFY THAT THE PATHOGEN IS INVASIVE ASPERGILLOSIS AND NOT SOME OTHER DISEASE WITHIN THE LUNG ENVIRONMENT. SO WHAT WE HAVE DONE IS WE USED THAT SAME ANTIBODY IMMUNOPET. SO WHAT WE DO, ALL OUR PRE-CLINICAL WORKS DONE IN MICE. WE IMMUNODEPLETE THOSE MICE AND MAKE THEME NEUTRO PENIC, THAT'S WHERE WE SEE THE DISEASE IN THE PATIENT. SO WE RENDER THEM NEUTRO PENIC, 24-HOURS LATER WE INJECT WITH SPEARS OF THE FUNGUS AND THE TRACER INTO THE TAIL. SO THE TRACER IS THE JF5 FULL-LENGTH ANTIBODY CONJUGATED WITH NEDAGGA TO COPPER 64 (?) SO THEN WE IMAGE 3 HOURS, 24 HOURS AND 48 HOURS POST-INJECTION AND CHALLENGE AND AT THE END OF 48 HOURS, EUTHANIZE THE ANIMALS AND MEASURE THE UPTAKE OF THE RADIO TRACER AND ORGANS EX-VIVO. SO WE ARE IMAGING UP TO THAT POINT IN-VIVO USING PET MRI. SO THE FIRST THING WE NEED TO DO WAS TO CHECK DOES IT ACTUALLY WORK? AND IT DOES. SO HERE WE HAVE MICE, NEUTRO PENIC MICE CHALLENGED WITH THIS AND YOU CAN SEE REALLY INTENSE UPTAKE OF THE TRACER WITHIN THE LUNG TISSUES. HERE WE HAVE MICE THAT HAVE A STERILE INFLAMMATORY TRIGGER. WE HAVE GOT NO UPTAKE WITHIN THE LUNG. STREPTOCOCCUS PNEUMONIA INFECTION WITHIN THE LUNG. NO TRACER UPTAKE AND ALSO INFECTION OF THE LUNG AGAIN MINIMAL UPTAKE COMPARED TO THE ASPERGILLUS. HERE WE HAVE QUANTITATIVE DATA. INJECTED DOSES OF LUNG AND SIGNIFICANT UPTAKE WITHIN THE LUNGS OF ASPERGILLUS INFECTED ANIMALS COMPARED TO THE OTHER ANIMALS. AND THIS IS EX-VIVO BY DISTRIBUTION WITH A GRAM OF TISSUE IN ORGANS AND YOU CAN SEE VERY NICE SPECIFIC SIGNIFICANT UPTAKE WITHIN THE LUNG TISSUE. SO THAT IS USING THE MOUSE JF5 ANTIBODY. WE WANTED THRONE TRANSLATE IT INTO THE CLINICAL SETTING FOR USE IN HUMANS AND TO DO THAT, WE HAVE TO DISGUISE THE FACT IT'S A MOUSE ANTIBODY. WHAT WE DID IS TO HUMANIZE THE MOUSE ANTIBODY BY GRAFTING THE CDR, HYPERVARIABLE LOOPS INTO AN IDG1 HUMAN FRAMEWORK. SO WE HAVE A HUMANIZE THE JF5 ANTIBODY THAT CONTAINS PREDOMINANTLY HUMAN PROTEIN AMINO ACIDS BUT THE HYPERVARIABLE LOOPS OF THE MOUSE BITS DO THE BONDING TO THE TARGET ANTIGEN. AND TO BE HONEST, WE GOT LUCKY. IT WORKED THE FIRST TIME WE DID IT. SOMETIMES WHEN YOU DO THIS, YOU CAN LOSE YOUR FUNCTIONALITY OF THE MOUSE ANTIBODY BY PUTTING IT INTO A HUMAN IGG1 FRAMEWORK. SO HERE WE HAVE PROOF OF THAT. WE HAVE OUR DEVELOPING SPORES OF ASPERGILLUS, PROBED WITH FLUORESCENT -- LABELED MOUSE ANTIBODY. YOU CAN SEE AGAIN WE HAVE GOT NICE DETECTION OF THE ACTIVE PART OF THE PATHOGEN AND HERE WE HAVE THE HUMANIZED GF5 ANTIBODY FICC LABELED. AGAIN WE RETAINED THE FUNCTION ATE AND DETECTION OF THE PARTS OF THE FUNGUS BUT NO DETECTION OF INACTIVE SPORES. THIS IS SHOW ARE PURITY OF THE HUMANIZED ANTIBODY AND THE ATTENTION OF BINDING IN A WESTERN BLOT TO THE GLYCOPROTEIN WHICH TYPICALLY IN FUNGI GIVE A HORRIBLE PROTEIN SMEAR. SO WE HAVE OUR HUMANIZED ANTIBODY. IN ORDER TO GET INTO HUMANS, WE HAVE TO DO REALLY EXTENSIVE RAT TOXICITY TESTING, PROBING OF HUMAN TISSUES TO MAKE SURE WE GOT NO NON SPECIFIC BINDING AND ALSO WE HAD TO IDENTIFY THE EPITOPE OF THE ANTIBODY. SO WHAT WE DID IS, WE DEVELOPED MUTANTS OF THE PATHOGEN AND WE HAD TARGETED BY HOMOLOGOUS COMBINATION, THE GENE AND ENCODING AND ENZYME AND WHAT THAT DOES IS ADDS ON RESIDUES TO FUNGAL GLYCOPROTEINS. WE HAD A SUSPICION THAT IT WAS RECOGNIZING GRAB TOES, THEY ARE VERY IMMUNOGENIC WITHIN FUNGI. SO WE DEVELOPED VARIOUS DIFFERENT STRAINS, KNOCKOUT STRAINS WHICH ARE DEFICIENT IN THE EPITOPE B215. SO HERE WE HAVE THE GENE KNOCKED OUT IN THE SOUTHERN BLOT SHOWING LOSS OF GENE WITHIN TWO INDEPENDENT MUTANT STRANDS. AND IN IMMUNOFLUORESCENCE WITH THE MOUSE GFI ANTIBODY, IN THE MUTANT STRAINS, WE LOST BINDING TO THE TARGET ANNUAL JEN. THE REALLY, REALLY WESTERN BLOTS AND ALSO WE DON'T GET ANY BINDING. SO WHAT WE WERE ABLE TO SHOW IS THAT THE ANTIBODY WAS RECOGNIZING THIS UPTAKE. THE IMPORTANT THING IS GALLIV IS ABSENT IN THE MAMMALIAN CARBOHYDRATES. SO WE WERE CONFIDENT WE WERE NOT GOING TO GET OFF SITE BINDING WHEN WE INJECTED INTO HUMANS. THERE IS A LOT OF IMAGES HERE. BASICALLY WHAT THIS IS SHOWING IS THE HUMANIZED JF564 COMPARED TO ALL SORTS OF DIFFERENT CONTROLS INCLUDING A HUMAN ISOTYPE CONTROL. AND IN INFECTED ANIMALS, WE GET REALLY NICE UPTAKE TRACER IN THE LUNGS OF INFECTED ANIMALS. AGAIN A HEAT MAP THERE IS SHOWING REALLY INTENSE UPTAKE WITHIN LUNGS. AND THE QUANTITATIVE DATA HERE SHOWING AGAIN SIGNIFICANT UPTAKE OF THE HUMANIZED ANTIBODY TRACER WITHIN THE LUGS OF ANIMALS INFECTED WITH THE PATHOGEN. SO THIS IS THE DON INSTRUCT WE HAVE TAKEN FORWARD FIRST-IN-HUMAN STUDIES THAT STARTED IF 2018. MORE RECENTLY, AND I'LL COME ON TO THAT IN A SECOND. MORE RECENTLY WHAT WE HAVE DONE IS SHOWN THAT YOU CAN USE THIS HJF5 TRACER TO MONITOR RESPONSE OF THE PATHOGEN IN-VIVO TO ANTIFUNGAL TREATMENT. SO WHAT WE HAVE HERE IS OUR ANIMAL MODEL, NEUTROPHIL DEPLETED ON DAY 0 INJECTED WITH A TRACER AND ALSO WITH A PATHOGEN INTO THE -- AND TREATED WITH A FRONT LINE ANTIFUNGAL TRY ANGAL DRUG USED IN THE CLINIC FOR CONTROLLING ASPERGILLOSIS AND THEN AGAIN INJECTED AT 24 HOURS AND THEN THE IN-VIVO STARTED. AND IT'S BEEN REPEATED 48 HOURS THE IN-VIVO IMAGING AND THEN THE ANIMALS ARE USED IN THE EX-VIVO BY DISTRIBUTION. WHAT THIS WORK HAS SHOWN IS BEING FOCUSED AND YOU NEED TO GET TREATMENT IN REALLY, REALLY EARLY WITH THE ANTIFUNGAL DRUG DRIVEN BY DIAGNOSIS. SO HERE WE HAVE ANIMALS ON ANIMAL LEFT-HAND SIDE HERE TREATED WITH -- AT 24 HOURS AND AT 24 HOURS YOU CAN SEE THE FUNGUS STARTING TO GROW WITHIN THE LUNG TISSUE AND THEN AT 48 HOURS IT'S COLONIZED THE LUNG. IF YOU TREAT IT FOR THREE HOURS AND THEN 24 HOURS, YOU CAN KNOCKOUT GROWTH AT THE PATHOGEN SIGNIFICANTLY REDUCING GROWTH OF THE PATHOGEN WITHIN THE LUNGS. AND HERE IS JUST THE QUANTITATIVE DATA. SO IT IS SHOWING PATH OF DIAGNOSTICS SHOWING IN-SITU AND IN-VIVO DIAGNOSIS AND RESPONSE TO TREATMENT WITH A FRONT LINE ANTIFUNGAL DRUG REALLY EXCITING. SO IN TERMS OF HUMANS, AS I SAID, WE STARTED A SMALL SCALE CLINICAL TRIAL. THE TRACE BEING ADMINISTERED TO PATIENTS ON COMPASSIONATE GROUNDS BUT OBVIOUSLY USING THEIR CONSENT. SO WE HAVE THREE PATIENTS. THEY ALSO HAVE ACUTE MYELOID LEUKEMIA. SO THEY ARE VERY HIGH RISK FOR FUNGAL INFECTIONS AS WELL AS OTHER LUNG INFECTIONS AND HERE WE HAVE A PATIENT DIAGNOSED WITH NO INVASIVE FORM OF ASPERGILLOSIS BASED ON CURRENT CLINICAL CONSENSUS DEFINITIONS OF DISEASE. AND THE RADIOLOGIST IDENTIFIED ABNORMALITY ON THE MRI. THE PATIENT WAS INJECTED AND THERE WAS NO UPTAKE OF THE TRACER WITHIN THE LESION IDENTIFIED FIRSTLY BY CT AND THEN MRI. WE HAVE A PATIENT WITH ACUTE INVASIVE PULMONARY ASPERGILLOSIS. A VERY DISTINCT LESION. WHEN INJECT WAD A TRACER, WE GET VERY INTENSE UPTAKE WITH THAT LESION. HERE WE HAVE A PATIENT WHOSE DISEASE DEVELOPED OVER SEVEN MONTHS SO THIS IS KNOWN AS SUBACUTE IPA. NOT LIKE THE VERY ACUTE THAT HAPPENS VERY, VERY QUICKLY. SO THIS PATIENT WAS IN FACT IMMUNOCOMPETENT. THEY WERE A TRANSPLANT PATIENT BUT WHAT HAPPENED IS THEY DEVELOPED THIS LESION ABNORMALITY OVERSEVEN MONTHS DESPITE ANTIFUNGAL TREATMENT AND THEN IT WAS DECIDED TO INJECT THE TRACER TO SEE WHETHER IT IS A INVASIVE FORM OF ASPERGILLOSIS. THE DIAGNOSIS WAS VERY, VERY DIFFICULT. IT WAS BASICALLY BASED ON SEROLOGY AND IDENTIFICATION OF ANTI-ASPERGILLUS IGG CIRCULATING WITHIN THE BLOODSTREAM. AND WHAT WE HAVE GOT IS AGAIN NICE UPTAKE WITHIN AN AREA WITHIN THAT ABNORMALITY. SO WE WERE ABLE TO CONFIRM THAT YES, THAT PATIENT HAD SUBACUTE IPA. WE IMAGED THREE PATIENTS. IT'S NOT AN EASY THING TO SET UP. YOU HAVE GOT TO HAVE SUSCEPTIBLE PATIENTS THAT ARE DIAGNOSED WITH THE DISEASE USING CONSENSUS DEFINITIONS AND HAVE EVERYTHING PRESENT WITHIN THE HOSPITAL IN ORDER TO DO THAT. BUT WE HAVE ACHIEVED IT, WHICH IS REALLY FANTASTIC. SO JUST IN SUMMARY, THE AS MERJILL US HAS BEEN TESTED FOR MANY YEARS AND SHOWED THAT THE BIOMARKER, GLYCOPROTEIN IS REALLY ABUNDANT WITHIN THE LUNG ENVIRONMENT T GAVE US CONFIDENCE WE COULD USE THAT SAME ANTIBODY PERHAPS IN RADIOLOGICAL DETECTION USING IMMUNOPETS AND MRI. SO WHAT WE HAVE DONE IS ALL THE PRE-CLINICAL IMAGING. WE HAVE NOW SUCCESSFULLY DETECTED DIFFERENT FORMS OF IN IPA IN PATIENTS WITH ACUTE MYELOID LEUKEMIA AND YOU CAN DO IMMUNOPET AND MRI ON FUNGAL INVASIVE DISEASES PROVIDED YOU HAVE A PATHOGENESIS MONOCLONAL ANTIBODY THAT BINDS TO ANTIGENS. AND AS PROOF OF THAT, WE HAVE DONE THAT WITH INVASIVE CANNED DID ISIS MANIFESTING AS KIDNEY DAMAGE, KIDNEY DISEASE WITH MULTIPLE LESIONS. WE HAVE A SYSTEM FOR MONITORING INVASIVE CANDIDIASIS USING A ANTIBODY AND WE GET NICE DETECTION WITHIN THE KIDNEYS OF INFECTED ANIMALS THAT HAVE A BLOODSTREAM INFECTION. SO THEY ARE IMMUNOCOMPETENT BUT HAVE A BLOODSTREAM INFECTION. SO I JUST LIKE TO FINISH NOW BY THANKING LOADS AND LOADS OF PEOPLE ON ALL THE IMAGING WORK DONE IN GERMANY AT THE UNIVERSITY OF -- AND MANY, MANY PEOPLE WHO HAVE BEEN INVOLVED IN THIS WORK ACROSS THE YEARS AND MANY DIFFERENT FUNDING AGENCIES. THE NIH FUNDED US DURING EARLY DEVELOPMENT STAGES OF THE FLOW DEVICE TO HAVE IT TESTED IN THEIR GUINEA PIG MODEL OF INVASIVE ASPERGILLOSIS AND I'D REALLY LIKE TO THANK THE EUROPEAN UNION WHO FUNDED US ALL ON A CONSORTIUM GRANT FOR FIVE YEARS TO TAKE THAT JF5 ANTIBODY AND GET INTO THE CLINIC WHICH WE HAVE DONE SUCCESSFULLY. SO THANK YOU FOR LISTENING AND THANK YOU TO ALL THE COLLABORATORS AND ALSO TO THE PATIENTS. WE WOULDN'T HAVE BEEN ABLE TO DO OUR WORK WITHOUT THEM. SO WE ARE EXTREMELY GRATEFUL TO THEM. THANK YOU. >> THAT WAS A GREAT TALK. WE ARE A FEW MINUTES OVER SO WE HAVE SOME QUESTIONS. I'D APPRECIATE IF ALL THE SPEAKERS FOR THE SESSION CAN COME ON VIDEO AND I'M GOING TO START ASKING SOME QUESTIONS. WE ARE GOING TO TRY TO TAKE AS MANY QUESTIONS AS WE CAN. WE HAVE ABOUT 5 MINUTES. THE FIRST QUESTION IS FOR DR. PETRIK. ARE THEY IS DERO FORES PART HOW DOES ONE SOLLUALIZE THEM AND THEN ONE OTHER QUESTION FOR YOU, THAT IS GALLIUM DFO TAKEN UP BY ACTIVATED IMMUNE CELLS? >> SO ACTUALLY I DIDN'T MENTION IN MY PRESENTATION BUT ALL WHICH WE USED ARE HIGHLY HYDROPHILIC SO NO PROBLEM TO CAN SOLUBLE IN WATER. AND REGARDING THE DFO AND THE IMMUNE CELLS, TO BE HONEST, WE NEVER TRIED SUCH AN ASSAY SO I SHOULD SAY, I DON'T KNOW. BUT BASED ON OUR RESULTS, ANIMAL RESULTS MAINLY AND MECHANISM HOW IT IS TAKEN UP BY BACTERIA AND FUNKY, I WOULD NOT EXPECT IT. >> THANK YOU VERY MUCH. I THINK THE ISSUE ABOUT SOLUBLIZATION IS PROBABLY ABOUT THE ONE THAT IS BOUND OR NOT BOUND TO THE METAL. I THINK THE UNBOUND STORM -- OR VICE VERSA. BUT WE CAN TAKE THAT QUESTION LATER. THANK YOU VERY MUCH DR. PETRIK. THE OTHER QUESTION IS FOR DR.-- WOULD THE SIDES OF THE VANCOMYCIN CONSTRUCT PROVIDE ACCESS IN CONNICK INFECTIONS WHEN THE ENDOTHELIAL BARRIERS ARE MORE OR LESS BACK TO NORMAL? ALSO IS THERE ANY NON-SPECIFIC ACCUMULATION OF AN VO MICE IN? >> THE FIRST QUESTION IS DIFFICULT FOR ME TO ANSWER TO BE HONEST. SO I DON'T KNOW. I GUESS WE JUST HAVE TO LOOK AT IT. THE VAN CO7 CAN BE USED TO TARGET BACTERIA INSIDE MACROPHAGES, SO IT DOESN'T SAY THAT MUCH. SO I WOULDN'T KNOW. BUT THE ACCUMULATION AT UNSPECIFIC SITES, WE DID OBSERVE SOME. BUT OF COURSE I SHOWED THE IMAGE WHERE THE VAN CO800 IN THE PLAIDER SO IT IS EXCRETED BY THE KIDNEY AND THE BLADDER YOU WILL SEE SIGNAL FROM THAT. AND IN ONE SET OF EXPERIMENTS WITH THE SPINE INFECTION THAT WE DID TOGETHER WITH ANOTHER LAB, WE ALSO SAW SOME OF THE MATERIAL LEAKING INTO THE ABDOMEN BUT WE DON'T KNOW WHY THAT IS THE CASE. IT'S A BIT ATYPICAL BUT IT HAPPENED. >> THANK YOU VERY MUCH. I HAVE A FEW MORE QUESTIONS BUT LET'S GO TO DR. THORNTON. GIVEN THAT SOME PATIENTS DEVELOP ANTI-ASPERGILLUS IGG, IS THERE POTENTIAL THESE BLOCK BINDING OF YOUR ANTIBODY BY COMPETING FOR THE ANTIGEN SITE? >> I GUESS IT'S A POSSIBILITY, YES. I GUESS IT'S ALL DOWN TO THE AFFINITY TO THE ANTIBODIES FOR THE TARGET AND THE HUMANIZED AND MOUSE ANTIBODY HAVE EXTREMELY HIGH AFFINITY SO THE POSSIBILITY -- I CAN'T SAY. BUT YOU WOULD HOPE THAT THE HIGH AFFINITY WOULD KNOCK OFF ANY ANTIBODY THAT IS ALREADY PRESENT. BUT IT'S IMPOSSIBLE TO SAY, TO BE HONEST WITH YOU. >> THANK YOU. AND I'M GOING TO ASK ONE MORE QUESTION WHICH IS OPEN TO EVERYBODY DOING ANTIBODY IMAGING. AND THE QUESTION IS ABOUT BOTH HOW WELL DO THEY PENETRATE POTENTIALLY NECROTIC LESIONS THAT WOULD HAPPEN WITH STAFF AUREUS AURAS PER JILL US? AND HOW DO YOU SEE -- I THINK IT'S AMAZING WHAT HAS BEEN SHOWN HERE WHERE STUFF HAS BEEN TRANSLATED TO THE CLINIC. HOW DO YOU SEE ANTIBODY-BASED TECHNIQUE BEING TRANSLATED TO THE CLINIC? WHAT ARE THE PITFALLS AND MAYBE SOME OF THE ADVANTAGES OF DOING THAT? THE PROCESS OF CLINICAL TRANSLATION? SO I DON'T KNOW IF DR. THORNTON YOU WANT TO TAKE IT OR DR. VAN JILL. >> SURE. TO BE HONEST WITH YOU, IT IS NOT TECHNICAL ISSUES THAT ARE THE PROBLEM IT'S MONEY AT THE END OF THE DAY. AND WHAT WE REALLY NEED IS A COMPANY WHO IS INVOLVED IN THAT KIND OF AREA TAKING THE TECHNOLOGY AND LICENSING AND USING THAT TECHNOLOGY. THAT'S ALL IT COMES DOWN TO. I MEAN, IT'S NOT TRIVIAL TASK PUTTING A HUMANIDES ANTIBODY INTO HUMANS. IT COSTS A LOT OF MONEY AND IT HAS TO BE PRODUCED UNDER CONDITIONS AND SO HAS THE COX 64 AND IT'S ALL GOT TO BE PUT TOGETHER UNDER THE CONDITIONS BEFORE YOU CAN PUT IT INTO A HUMAN. HAVING SAID THAT, GIVEN THE AMOUNT OF MONEY THAT IS SPENT ON HUMANS WITH INVASIVE FUNGAL DISEASES, THE POTENTIAL SAVING IS HUGE COMPARED TO GIVING THEM THE DRUG. SO IF YOU CAN SHOW THAT DON'T HAVE THE DISEASE THEN YOU CAN WITH HOLD THE DRUG RATHER THAN LENGTHY HOSPITAL STAYS WITH UNNECESSARY TREATMENT. SO IT'S JUST DOWN TO MONEY TO BE HONEST. >> THANK YOU. I THINK PEOPLE WOULD -- OR AT LEAST THE PERSON ASKING THE QUESTION IS HOPING THAT PEOPLE CAN SHARE THEIR EXPERIENCES SO THAT OTHERS WOULD DEVELOP THESE PATHWAYS. SO BASICALLY I'M GOING TO ASK THEM TO CONTACT YOU DIRECTLY SO THAT THEY CAN UNDERSTAND THE PATHWAY. DR. VAN JILL DO YOU WANT TO QUICKLY TRY TO ANSWER THE QUESTION ABOUT THE PENETRATION OF THE ANTIBODY INTO POTENTIALLY NECROTIC PLACE? >> SO WE HAVE ONLY EXAMPLES OF MOUSE EXPERIMENTS BUT THE ANTIBODIES WERE HIGHLY SPECIFIC AND AGAIN REFERRING TO THE SPINAL IMPLANT INFECTION WHEN WE SAW IS THAT OVER LONG PERIODS OF IMAGING INFECTION FOR SPECIFICITY AND IT WOULD LEAK AWAY A LITTLE BIT AND MONOCLONAL ANTIBODY WOULD STAY FOR SEVERAL DAYS. SO WHAT WE DID, WE HAD A SLIGHTLY DIFFERENT APPROACH SO WE CLONED THE ANTIBODY DIRECTLY FROM THE B CELLS SO WE DIDN'T HAVE TO HUMANIZE. WE HAD SEVERAL AND THEN THEY WERE TAKEN FROM PATIENTS WHO HAVE -- GENETIC DISEASE THAT CAUSES CHRONIC WOUNDS AND THEY -- NOT ALL THE PATIENTS BUT MANY HAVE LONG TERM STAFF LA CAULKAL COLINNIZATION WITH MULTIPLE -- THEY WERE KIND ENOUGH TO MAKE B CELLS AND I THINK ANTIBODIES ARE GREAT BUT THEY ARE BIGGER THAN THE VANCOMYCIN. I DON'T KNOW. >> EXCELLENT. I THINK TIME WILL TELL. SO FANTASTIC PRESENTATION AGAIN. I WANT TO THANK ALL THE SPEAKERS. WE ARE GOING TO TAKE A BREAK I'M A RESEARCH FELLOW AT JOHNS HOPKINS. WE HAVE FOUR SPEAKERS. THE TALKS WILL BE CONTINUOUSLY PRESENTED AND THERE WILL BE A DECISION SESSION AT THE END. PLEASE REMEMBER TO WRITE YOUR QUESTIONS IN THE CHAT. OUR FIRST SPEAKER IS DR. HENRY VANBROCKLIN A PROFESSOR OF RADIOLOGY AT THE UNIVERSITY OF CALIFORNIA SAN FRANCISCO. THE TITLE OF HIS TALK IS IMAGING HIV INFECTION SEARCHING FOR THE RESERVOIR. >> CAN YOU SEE MY SLIDES AND HEAR ME OKAY? >> YES. WOULD YOU MIND SWAPPING TO DISPLAY? >> HOUSE THAT? >> PERFECT. >> GREAT. THANK YOU VERY MUCH FOR THE KIND INTRODUCTION AND I'D ALSO OF COURSE LIKE TO THANK YOU FOR INVITING ME TO SPEAK TODAY. I HAVE WATCHED MOST OF THE PRESENTATIONS. I GOT UP AT 5:30 THIS MORNING TO JOIN THE FIRST SESSION AND FOUND THESE TO BE VERY INTERESTING SET OF PRESENTATIONS AND I THANK THE ORGANIZERS FOR PULLING THIS TOGETHER. YOU CAN SEE THE BACKDROP ON MY SLIDE IS A LITTLE BIT OF PLAY ON WORDS, SEARCHING FOR THE WES VOYEUR. LIVING HERE IN CALIFORNIA, WE ARE UNDERGOING A SIGNIFICANT DROUGHT AND THE PICTURE ON THE RIGHT-HAND SIDE OF THE SLIDE IS A REAL PICTURE TAKEN THIS YEAR OF SOME OF OUR RESERVOIRS AND LAKES HERE IN CALIFORNIA. SO WE ARE ALSO SEARCHING FOR OUR OWN RESERVOIRS OF WATER SO WE CAN MAINTAIN OUR QUALITY OF LIFE. LET'S SHIFT TO DISCUSS THE TOPIC AND THAT IS, HIV AND INFECTION, IMAGING THE INFECTION. LAST WEEK, WE COMMEMORATED THE 40th ANNIVERSARY OF THE FIRST PATIENTS THAT WERE SEEN IN THE HOSPITALS AND DOCTOR'S OFFICES WHO HAD CONTRACTED WHAT WE KNOW TODAY AS HIV AND AIDS. AND MUCH LIKE WHAT HAPPENED OVER THE LAST YEAR AND A HALF WITH COVID, THIS STARTED, OF COURSE, A EFFORT AMONG A LARGE GROUP OF PEOPLE ACROSS THE GLOBE TO TRY AND FIND CURATIVES FOR HIV OR FIND AT LEAST TREATMENTS THAT WOULD SLOW THE PROGRESSION OF THE DISEASE AND WOULD PRESERVE THE QUALITY OF LIFE FOR THOSE PATIENTS THAT CONTRACTED HIV OR AIDS. OF COURSE AS WE ALL KNOW NOW, 40 YEARS LATER, WE ARE NOT ABLE TO CURE HIV TODAY. BUT WE ARE ABLE TO STOP THE SPREAD OF THE DISEASE TO AN EXTENT AND THIS DISEASE, I SHOULD MENTION, CLAIMED OVER 30 MILLION LIVES WORLDWIDE AND IN THE U.S., CLOSE TO -- A LITTLE MORE THAN 700,000 LIVES. SO THIS IS AN ATTRACTABLE DISEASE BUT FORTUNATELY, DUE TO THE HARD WORK AND EFFORT OF FOLKS IN THE EARLY 80s, THEY WERE ABLE TO FIND AN ANTI-RETROVIRAL THERAPY A REMARKABLE TREATMENT, WHICH IS ABLE TO KNOCK THE VIRUS DOWN, HELP TO PREVENT THE SPREAD OF THE VIRUS FROM ONE INDIVIDUAL TO THE NEXT AND TO REALLY PROVIDE A HIGH-QUALITY OF LIFE FOR THOSE THAT HAVE CONTRACTED AIDS. AND THERE ARE SEVERAL PATIENTS FROM THE MID TO LATE 80s AFTER AZT WAS FIRST CAME ON THE MARKET THAT ARE STILL AROUND TODAY AND IT IS -- OR SEVERAL PATIENTS FROM THE LATE 80s OR EARLY 90s STILL AROUND TODAY. SO IT'S REMARKABLE WE HAVE THIS KIND OF TREATMENT. BUT WE NEED TO BE ABLE TO HELP TO DEVELOP A CURE. THAT'S REALLY WHERE WE ARE HEADED. SO WE KNOW THAT INFECTED CELLS PERSIST DESPITE ANTI-RETROVIRAL TREATMENT SO IT'S A LIFELONG ANTI-RETROVIRAL THERAPY FOR MOST. THERE ARE SOME CONTROLLERS, PEOPLE THAT CAN ACTUALLY CONTROL THEIR DISEASE, SOMEWHAT NATURALLY. SO THEY DON'T REQUIRE ANTI-RETROVIRALS. BUT FOR A MAJORITY OF THE PATIENTS THEY WILL EXPERIENCE A MANIFESTATION OF THE DISEASE AND HAVE TO BE ON ART LIFELONG. SO THE HIV RESERVOIR IS A MAJOR CURATIVE HURDLE AND ONE OF MY COLLEAGUES WHO I WORK WITH, DENNIS BECK, WHO DID A LOT OF THE WORK ON THIS DEVELOPMENT, LIKENED THIS TO TRY TO FIND WALL DOE IN THIS. WALL DOE IS WEARING A STRIPED SHIRT. WE CAN IDENTIFY HIM BY THAT STRIPED SHIRT BUT TRYING TO FIND THE RESERVOIR IS LIKE TRYING TO FIND WALL DOE IN THIS PICTURE. OVER THE LAST DECADE OR SO, THERE HAS BEEN A RENEWED EFFORT TO TRY FOR A CURE. THERE ARE OVER 30 DRUGS NOW THAT ARE USED TO SORT OF TAME THE PATIENTS IN HAVE THE DISEASE, HIV, BUT THE PERSISTENCE IN THE SEARCH FOR A CURE IS REALLY CRITICAL. SHOWN HERE ARE TWO PAPERS FROM THE LAST DECADE, WHICH SAY THAT ONE OF THE WAYS THAT WE ARE GOING TO BE ABLE TO COME UP WITH THIS CURE IS GOING TO BE INVOLVED, UNDERSTANDING WHERE THE RESERVOIR IS AND WHERE THE PERSISTENT HIV RESIDES, WHERE THE VIRUS RESIDES IN THE BODY. SHOWN HERE THAT WE NEED TO HAVE MODELS AND ALSO TO BE ABLE TO TRANSLATE THIS INTO HUMANS SO WE CAN FIND THE CELLULAR AND TISSUE SOURCES OF THE PERSISTENT HIV, EVEN ON THOSE INDIVIDUALS THAT ARE ON LONG TERM ANTIRETROVIRAL THERAPY. TO DEVELOP THERAPEUTICS AND TO BE ABLE TO FOLLOW WHAT IS HAPPENING TO THE RESERVOIR AS THESE THERAPEUTICS ARE BEING GIVEN. AND SO IT WAS WORKING WITH MY COLLEAGUES IN THE LATE THOUSANDS AND FUNDING BY THE AMFA, AMERICAN FOUNDATION FOR AIDS RESEARCH, THAT SPONSORED A BIG PUSH TO TRY AND FIND A CURE, THAT WE EMBARKED ON A DEVELOPMENT OF IMAGING AGENTS THAT COULD HELP US TO PERHAPS UNDERSTAND WHERE THE RESERVOIR WAS. WE WERE INSPIRED BY WORK THAT WAS DONE IN THE MID 2010s BY SANITAGE LOW AND HIS GROUP, WHERE HE TOOK A RADIOLABELED ANTIBODY, AND INJECTED THIS INTO A SIMIAN IMMUNOVIRUS MODEL, A PRIMATE THAT HAS THE PRIMATE VERSION OF AIDS. YOU CAN SEE HERE IN THE VIREMIC IN PANEL A, THERE WAS SIGNIFICANT UPTAKE IN DEBTBUTION OF THE LABELED ANTIBODY THROUGH AN ENVELOPE COAT PROTEIN. YOU CAN SEE UPTAKE IN THE NASAL AREA. YOU CAN SEE UPTAKE IN THE GUT AND LYMPH NODES THAT THEY WERE ABLE TO TEASE OUT FROM THIS IMAGING, AS COMPARED TO A NORMAL HEALTHY CONTROL WHERE YOU SEE VERY LIMITED DISTRIBUTION OF THIS MATERIAL. AND THEY WERE EVEN ABLE TO IDENTIFY THE REDUCTION OF THE DISEASE BY TREATING THESE ANIMALS WITH ANTI-RETROVIRALS. SO THIS WAS SORT OF A STIMULUS FOR US TO TRY AND DO THIS AND DO THIS IN HUMANS. IT'S VERY CHALLENGING TO DO HIV WORK BECAUSE IT DOES INVOLVE VERY HIGH-CLASS LABORATORIES TO CONTAIN THIS WORK SO IT'S VERY CHALLENGING TO DO STUDIES IN ANIMALS THAT HAVE THE DISEASE SO THERE IS ONLY CERTAIN LABORATORIES IN THE STATES AND ALSO IN THE WORLD THAT CAN HANDLE THIS TYPE OF RESEARCH. AND NORMALLY, OF COURSE, WE WOULD USE DISEASE MODELS AS YOU HAVE SEEN TODAY THROUGHOUT MANY OF THE TALKS IN MICE AND OTHER ANIMAL MODELS, TO STUDY AND EVALUATE OUR TRACERS. FORTUNATELY FOR US, THE VACCINE RESEARCH CENTER AT THE NIH HAD DEVELOPED SOME BROADLY NEUTRALIZING ANTIBODIES THAT BOUND TO THE CD4 BINDING SITE PROTEIN GP120 ON THE VIRAL COAT FOR THE -- OR THE VIRAL COAT FOR THE ANTIBODIES. THESE NEUTRALIZING ANTIBODIES, AND WE HEARD SOME OF THIS DISCUSSION FOR COVID, IS A WAY OF BINDING THE ANTIBODY TO THE ANTIGEN AND TO STOP AND PREVENT THE DISEASE FROM PROLIFERATING. WHAT I SUMMARIZED ON THIS SLIDE IS ABOUT A YEAR AND A HALF'S WORTH OF WORK WHERE WE RECEIVED THE GMP ANTIBODY FROM VACCINE RESEARCH CENTER. THE GMP ANTIBODY HAD ALREADY BEEN STUDYING IN HUMANS AS A POTENTIAL TO GO AFTER HIV AND WE WERE ABLE TO TAKE THIS, CONJUGATE THE DFO, LABEL WITH ZIRCON YUM 89 AND SHOW WE MAINTAINED THE IMMUNOREACTIVITY OF THE MATERIAL AND DID IMAGING IN SMALL ANIMALS, NOT WITH HIV. THESE WERE NORMAL ANIMAL. I WILL SHOW AN IMAGE IN A FEW MINUTES. WE DID A PRIMATE BUT NOT HIV PRIMATE. AND THEN WE WERE ABLE TO TRANSLATE THIS, GET THE REGULATORY PAPERWORK AND GET PERMISSION FROM THE FDA TO GO AHEAD AND MOVE THIS INTO HUMANS AND WERE ABLE TO DO HUMAN IMAGING. AT THE SAME TIME, AT UC DAVIS, THEY WERE DEVELOPING THE TOTAL BODY PET AND THIS IS THE SMALL ANIMAL VERSION OF THE TOTAL BODY PET. WE ACTUALLY WERE ABLE TO DO SOME OF THE FIRST IMMUNOPET IMAGING STUDIES USING THE TOTAL BODY PET INSTRUMENT AT UC DAVIS IN THE PRIMATES. THESE ARE NORMAL PRIMATES. THIS IS INJECTION OF ABOUT A MILL CURRY OF THE ZIRCON YUN 89 AND UP TO SIX DAYS LATER, TWO HALF LIVES, YOU CAN STILL SEE THE DISTRIBUTION. THIS IS NORMAL DISTRIBUTION, NORMAL PHARMACOKINETICS, A LOT OF BLOOD POOL HERE IN THE EARLY IMAGE ON THE FIRST DAY AND BY SIX DAYS YOU SEE MOST OF IT IN THE LIVER. YOU SEE A LITTLE BIT OF KIDNEYS. STILL SOME BLOOD POOL AND YOU NOTICE THERE IS AGENT TAKEN UP IN THE JOINTS WHICH MAY SOMEBODY FREE ZIRCON YUM IN THIS PARTICULAR MODEL. THIS IS NOT SOMETHING WE SEE -- SO WE TRANSLATED THIS NOW INTO OUR FIRST-IN-HUMAN SUBJECTS USING PET MR. SHORTLY AFTER WE HAD GOTTEN APPROVAL FROM THE FDA TO DO THIS, AND YOU CAN SEE THIS IS SORT OF OUR FIRST COUPLE OF IMAGES, ONE ON THE LEFT FROM HIV NEGATIVE CONTROL AND ONE OF THE RIGHT FROM HIV POSITIVE OR VIREMIC PATIENT. THIS PATIENT IS ACTUALLY ONE THAT HAD ACTIVE DISEASE, WAS NOT ON ANTI-RETROVIRALS AT THE TIME THAT WE TOOK THESE IMAGES. AGAIN YOU CAN SEE VERY SIMILAR TO WHAT WE SAW IN THE EARLY TIME POINT FOR THE PRIMATES THAT YOU CAN SEE THE BLOOD POOL AND JUST NORMAL DISTRIBUTION HERE. YOU DO SEE A LITTLE BIT OF UPTAKE IN THE HIV NEGATIVE CONTROLS. YOU SEE CONSIDERABLE AMOUNT IN THE BOWEL OF THE HIV SUBJECTS. BUT THIS ISN'T THE ENTIRE STORY. YOU REALLY HAVE TO DIG DOWN INTO THE IMAGES TO BE ABLE TO SEE WHERE THIS UPTAKE IS RESIDING. SO I SHOWED HERE THREE IMAGES, ONE FROM A VIREMIC PATIENT ON THE LEFT. THE MIDDLE ART SUPPRESSED PATIENT, UNINFECTED. THIS IS LYMPH NODES IN THE GUT RIGHT AROUND THE PELVIS AREA AND YOU CAN SEE HERE QUANTITATIVELY IN THE FIVE SUBJECTS WE STUDIED, FIVE OF EACH, VIREMIC, ART SUPPRESSED AND UNINFECTED CONTROLS. EVEN AS EARLY AS THE FIRST DAY WE ARE STARTING TO SEE DIFFERENTIATION IN UPTAKE BETWEEN THE CONTROLS AND THE VIREMIC PATIENTS AND WHAT IS EVEN WONDERFUL TO SEE IS HERE AT DAY 3, WE CAN SEE A DIFFERENCE BETWEEN THE CONTROLS AND ART AND YOU CAN SEE THERE IS A DECREASE STARTING TO FORM. WE JUST HAVEN'T DONE ENOUGH PATIENTS TO MAKE THAT SIGNIFICANT BUT THERE IS A DECREASE IN THE ART SUBJECTS. SO THIS IS A SIMILAR PATTERN AND WE LOOKED AT THEM TO DAY SIX AND NOW WE ARE STARTING TO SEE THE PERHAPS SIGNIFICANT DIFFERENCES BETWEEN OUR SIGNALS, CONTROL PATIENTS VERSUS VIREMICS AND OUR NORMAL PATIENTS. JUST HAVEN'T DONE MUCH SUBJECTS TO GET STATISTICAL SIGNIFICANCE. WE ALSO NOTED UPTAKE IN OTHER AREAS WHERE HAD NOT NECESSARILY BEEN IDENTIFIED BEFORE IN PATIENTS, POTENTIAL PLACES WHERE THE RESERVOIR MAY BE. THE BIGGEST ONE OF THOSE WAS IN BONE MARROW. HERE YOU CAN SEE THIS IS IN SPINE. YOU CAN SEE UPTAKE IN THE VIREMIC, ART SUPPRESSED AND LITTLE IN THE UNINFECTED. HERE IS PELVIC MARROW. WE CAN SEE A DIFFERENCE HERE BETWEEN THE CONTROLS AND THE VIREMICS AND THE PELVIC MARROW. NASAL TERMINATES, LYMPHOID TISSUES IN THE NASAL AREA UP IN THE HEAD AND WE ARE ALSO SEEING SOME DIFFERENCES IN THE UPTAKE FROM CONTROLS TO ART AND VIREMIC PATIENTS IN THE GUT WOO DID INITIAL STUDIES TAKING LYMPH NODE BOPSITIES TRY TO START TO VALIDATE THESE DATA AND SHOWN HERE IS THE UPTAKE LEVELS IN THE LYMPH NODE WITH THE MEASUREMENT OF P24 PLUS IN THE LYMPH NODE TISSUES. THIS IS A MARKER OF THE VIRUS IN THESE TISSUES AND YOU CAN SEE HERE THERE IS A GOOD CORRELATION EVEN WITH THE FIRST FEW PATIENTS THAT WE STUDIED AND WERE ABLE TO BIOPSY. SO WE WILL BE ABLE TO DO SOME WORK IN TERMS OF VALIDATING THAT WHAT WE ARE SEEING IN TERMS OF UPTAKE WITH THESE AGENTS IS ACTUALLY CONSISTENT WITH THE VIRAL RESERVOIR. WHAT IS ALSO INTERESTING IN THE FIRST FEW ART SUPPRESSED PATIENTS, WE LOOKED TO THE UPTAKE VERSUS THE TIME THEY HAD BEEN ON ART AND YOU CAN SEE OUT TO 10,000 DAYS, OVER 27 YEARS, AND YOU SEE THIS NICE DECREASE SO FAR A PERIOD OF TIME. AGAIN THIS IS ANECDOTAL. WE NEED MORE PATIENTS MAKE THIS SIGNIFICANT BUT WHAT IS ALSO INTERESTING TO NOTE IS THAT THIS VALUE HERE FOR THE LYMPH NODE IS STILL ABOVE WHAT WE SEE FOR THE CONTROL SUBJECTS. WE REALIZE THAT IN THE HUMAN CASE N THIS PARTICULAR CASE, WE DON'T HAVE ANY ENDOGENOUS GP120 ANYWHERE IN OUR BODY. THE ONLY PLACE WHERE IT IS COMING IN HAS TO BE FROM THE VIRUS ITSELF. AND SO, THIS IS ANOTHER REASON THITHIS IS A GREAT DISEASE TO STUDY BECAUSE THEY HAVE A VERY LOW BACKGROUND IN TERMS OF THE MARKERS, THE VIRAL MARKERS THAT WE ARE ABLE TO SEE INSIDE OF THESE HUMAN SUBJECTS. AGAIN TAKING ADVANTAGE OF THE FACT THAT THE TOTAL BODY PET WAS INSTALLED AT UC DAVIS IN OR THE MIDDLE OF 2019, WE WERE ACTUALLY ABLE TO USE THIS HIGH SENSITIVITY IMAGING SCANNER TO LOOK AT A COUPLE OF THE PATIENTS THAT WE HAD INJECTED WITH THE ZIRCON YUM 89-LABELED ANTIBODY. AND SHOWN HERE IS THE PARADIGM THAT WE USED. WE INJECTED THE PATIENT AT UCSF. WE IMAGED THEM IN THE MORNING ON THE PET MR AND THEN DROVE THEM IN A CAR, THE 70 MILES BETWEEN UCSF AND UC DAVIS MEDICAL CENTER, WHICH IS ACTUALLY LOCATED IN SACRAMENTO. YOU CAN SEE HERE ONE HOUR AND 6 MINUTES, THAT'S A VERY LIGHT TRAFFIC DAY. THIS NORMALLY WOULD TAKE ABOUT 90 MINUTES TO TWO HOURS TO GET BETWEEN THE TWO PLACES. AND THEN WE WERE ABLE TO STUDY THE PATIENT ON THE EXPLORER. AND HERE ARE THE FIRST IMAGES WE RECEIVED. YOU CAN SEE HERE ON THE LEFT THE UPTAKE FROM THE PET MR. THIS IS A MULTIPLE-BED POSITION AT 5 MINUTES FOR EACH BED POSITION, SO 6 BED POSITIONS T TOOK US 30 MINUTES TO GET THIS IMAGE ON THE LEFT T TOOK 20 MINUTES FOR US TO GET THE HEAD-TO-TOE IMAGE FROM THIS PATIENT AND THIS IS 20 MINUTES AT EVERY POINT HERE. NOT JUST 5 MINUTES AT EACH POINT AS YOU CAN SEE ON THE LEFT. HERE IS AN HIV PATIENT. THIS WAS DONE 72 HOURS POST-INJECTION AND WE HAD A SECOND PATIENTS CAME IN A DAY LATER AND GOT INJECTED WITH VALID YUM, ANTIBODY, AND THIS WAS A PATIENT WHO WAS ON ART. AND YOU CAN SEE VERY SIMILAR UPTAKE BETWEEN THE TWO AND OF COURSE YOU CAN'T SEE ALL THE DETAILS HERE ON THESE PROJECTIONS BUT WE DID EVALUATE THESE QUITE CLOSELY. YOU CAN SEE UPTAKE IN THE GUT WHICH IS PRETTY CONSISTENT AND ALSO UPTAKE IN THE MARROW, WHICH WE ARE SEEING HERE IN THIS. AGAIN, LOOKING CLOSELY AT THE DISEASE, WE SAW UPTAKE IN THE LYMPH NODES AND IN THE MARROW AND WE WERE ABLE TO CORROBORATE THE DATA BETWEEN THE PET MR AND THE EXPLOREY PET, EXPLORER BEING MORE SENTTIVES THAN THE PET MR. I KNOW I'M RUNNING OUT OF TIME. I'D LIKE TO TRANSITION QUICKLY TO TALK ABOUT IMAGING T CELL ACTIVATION. WE HAVE BEEN WORKING FOR SEVERAL YEARS TO HELP THEM TRANSLATE A MOLECULE THAT WAS DEVELOPED BY ANOTHER GROUP TAKING THIS INTO HUMAN IMAGING. WE DID THE FIRST HUMAN IMAGING SUBJECTS IN ABOUT 2015 AND THEY WERE ABLE TO SHOW THAT FARAG GETS TAKEN UP INTO ACTIVATED T CELLS AND THEY HAVE DONE THIS IN HUMANS AND ANIMALS. WHAT WE HAVE BEEN ABLE TO DO WITH PET MR IMAGING IS SHOW WE ARE SEEING CONSISTENT UPTAKE IN AREAS WHERE WE SEE THE HIV INFECTION HERE IN THE NASAL TERMINATES. AND ALSO HERE IN THE LYMPH NODES AND ALSO IN THE MARROW WE SEE. SO AGAIN JUST LOOKING ACROSS BETWEEN THE HIV PATIENTS, BOTH ON ART. ALL PATIENTS TAKEN TOGETHER SOME ON PART AND SOME ARE VIREMIC, WE CAN SEE INCREASE IN UPTAKE AND VERY LOW UPTAKE IN THE CONTROL OR HEALTHY CONTROL. SO WHERE DO WE GO FROM HERE? WE PLAN ON INTRODUCING THIS AND USING THIS ON STUDIES WITH TREATMENT INTERRUPTION AND ALSO ON EXISTING TRIALS TO LOOK AT NEW CURES. AND FINALLY, TO JUST SUMMARIZE WE DEFINITELY IN THIS LABELING HAVE BEEN ABLE TO SUCCESSFULLY LABEL THE BRCL1 -- VRC01. STRONG CORRELATION OF THE LYMPH NODE UPTAKE BETWEEN THE P24 MEASURES OF HIV INFECTION CORRELATION BETWEEN THE LYMPH NODE AND YEARS ON ART IS VERY INTERESTING AND CORROBORATED SOME OF THE PRE-CLINICAL STUDIES THAT WE ARE SEEING IN THE PRIMATES. AND IN WORK THAT IS STILL ONGOING, AND WILL CONTINUE NOW TO ACCRUE MORE PATIENTS IN BOTH ARMS HERE, BUT WE SEE INCREASED UPTAKE IN GLANDULAR LYMPHOID TISSUES WHERE THE T CELL ACTIVATING AGENT AND INCREASED UPTAKE IN THE BONE MARROW AND WE ARE STARTING TO SEE CORRELATION BETWEEN SOME OF THESE EMERGING CORRELATIONS BETWEEN SOME OF THESE TISSUES. AND THE IMAGING OF THESE TWO AGENTS RIGHT NOW MAY HELP US TO CHARACTERIZE THE RESERVOIR AND BE INCORPORATED IN CLINICAL TRIALS TO HELP US ASSESS FUTURE THERAPEUTIC INTERVENTIONS AND HOPEFULLY MOVING TOWARDS THE CURE. IT DOES TAKE A VILLAGE AND I'D LIKE TO ACKNOWLEDGE THE SUPPORT FROM THE AMERICAN FOUNDATION FOR AIDS RESEARCH, NIH, THROUGH A DARE COLLABORATORY HELD BY STEVE DEACS SHOWN HERE AND ALSO A RECENT RO1 WE GOT TO CONTINUE THESE STUDIES. THIS IS TIM HEN RICH IN THE LOWER RIGHT WHO IS A PHYSICIAN AND TREATS HIV PATIENTS AND IS A WONDERFUL COLLABORATOR AND HAS BEEN WORKING WITH HIM FOR SEVERAL YEARS TO MAKE THIS HAPPEN. AND THE REST OF THE TEAM THAT MAKES THIS HAPPEN AND CELL SITE TECHNOLOGIES FOR THEIR VRC01 VACCINE RESEARCH. WITH THAT, I THANK YOU. >> THANK YOU VERY MUCH THAT WAS A REALLY IMPRESSIVE TALK. IT IS NOW WITH GREAT PLEASURE I INTRODUCE OUR NEXT SPEAKER, DR. DIMA HAMMOUD IS A SENIOR INVESTIGATOR AND DEPUTY DIRECTOR FOR THE CENTER OF DISEASES IMAGING AT THE NIH. HER TALK IS MOLECULAR IMAGING OF HIGH CONSEQUENCE VIRAL INFECTION INFECTION. >> THANK YOU VERY MUCH FOR THAT INTRODUCTION AND THANK YOU FOR -- YOU WILL JUST -- I THINK YOU'RE SHARING THE WRONG SCREEN. >> THANK YOU VERY MUCH FOR THE INTRODUCTION AND THANK YOU ALL FOR STICKING AROUND WITH US THROUGH THIS LONG DAY OF TALKS. I HAVE TO SAY IT'S BEEN AMAZING, VERY IMPRESSIVE. SO, OVER THE NEXT 20 MINUTES OR SO I'LL BE TALKING ABOUT MOLECULAR IMAGING OF HIGH CONSEQUENCE VIRAL INFECTIONS WITH A SPECIFIC FOCUS ON EBOLA VIRUS. AS YOU ALL KNOW, HIGH CONSEQUENCE VIRAL INFECTION ARE DEVASTATING AND WE KNOW THEY ARE RESPONSIBLE FOR MULTIPLE EPIDEMICS. THE ONES ARE THE 2014 EPIDEMIC OF EBOLA IN WEST AFRICA AND COVID-19 WHICH WE ARE STILL RECOVERING FROM. AS WELL AS SMALLER ONES LIKE ZIKA, AND WREAK HAVOC. THE PROBLEM WITH MANY OF THOSE IS THAT THEY OCCUR IN UNDER DEVELOPED COUNTRIES AND BECAUSE OF THAT, THERE IS NO INFRASTRUCTURE TO TRY TO BETTER UNDERSTAND THE PATHOPHYSIOLOGY OF DISEASE IN THOSE SITUATIONS. SO THE BEST OR NEXT BEST THING TO DO IS TO USE ANIMAL MODELS AND WITH IMAGING, WE CAN DEVELOP NON-INVASIVE BIOMARKERS OF DISEASE. THE GOALS WOULD BE TO BETTER UNDERSTAND THE PATHOPHYSIOLOGY OF THE DISEASE AS WELL AS THE DYNAMICS AND TO EVALUATE THE EFFICACY OF VARIOUS TREATMENTS AND VACCINES. SO TODAY I WILL BE TALKING ABOUT TWO MAIN PROJECTS WE HAVE DONE, THE FIRST ONE WE USE A LIGAND ALONG WITH HISTOPATHOLOGY IN A MONKEY MODEL -- [ READING ] WE ALSO HAVE ANOTHER PROJECT WHERE WE ARE LOOKING MORE CAREFULLY AT TNF INVOLVEMENT IN THE BRAIN AND EBOLA FOR WHICH WE USE FDG PET AND MRT1. SO ALL OF THIS WORK HAS BEEN DONE BY THE INTEGRATED RESEARCH FACILITY IN FREDRICK, MARYLAND. THIS THE RESEARCH FACILITY YOU WILL HEAR MORE ABOUT IN THE NEXT TALK. IT'S AN EXCELLENT FACILITY WHICH WAS BUILT SPECIFICALLY TO MANAGE, COORDINATE AND FACILITATE THE CONDUCT OF RESEARCH ON THE EMERGING INFECTIOUS DISEASES AND BIODEFENSE PATHOGENS AS NEEDED. IT'S A COLLABORATIVE FACILITY WITH FOCUS ON ADVANCED IMAGING AND THE BEST PART IS THAT IT'S AVAILABLE TO THE SCIENTIFIC COMMUNITY FOR COLLABORATIVE PROJECTS. SO THE ANIMAL MODEL WE WORK ON IS ACTUALLY HAS BEEN WORKED ON AND DEVELOPED AND THERE ARE LOTS OF STUDIES DONE AT IRF ON THIS MODEL. IT'S VERY GOOD SIMULATION OF THE DISEASE. IT'S VERY SIMILAR PROGRESSION TO HUMAN DISEASE BUT THE DISEASE FORCE IS A LITTLE BIT SHORTER. GENERALLY FOLLOWING KNOCKALATION WITH THE VIRUS EBOLA VIRUS TARGETS THE CELLS AND MONOCYTES AND MACROPHAGES AND EVENTUALLY SPREAD TO LYMPH NODES AND THEN TO BLOODSTREAM AROUND DAY 3 OR 4 AND EVENTUALLY REACHING OTHER ORGANS. THE TYPICAL MANIFESTATIONS INCLUDE MONOCYTIC AND LYMPHOCYTIC APOPTOSIS AND THE LAST PART OF THE DISEASE INCLUDES A CYTOKINE STORM FOLLOWED BY MULTIORGAN FAILURE, SHOCK AND DEATH. THERE HAS BEEN A LOT OF WORK DONE ON EBEALE ON VIRUS AS WELL AS OTHER VIRUSES AT IR. AND THIS IS A STUDY THAT WAS DONE BY DR. JOHNSON AND THE GROUP AT IRF WHERE ANIMALS WERE EVALUATED WITH FDG PET. THESE ARE IMAGES OF THE WHOLE ANIMAL AND THEN MULTIPLE TIME POINTS AFTER KNOCKALATION. THERE WAS EVIDENCE OF INCREASED UPTAKE IN THE SPLEEN AS WELL AS IN THE BONE MARROW AND MULTIPLE LYMPH NODES AS WELL AS THE GI TRACT. SO BASED ON ALL THE PREVIOUS WORK THAT HAS BEEN DONE, WE DECIDED THAT WE WOULD LIKE TO LOOK MORE CAREFULLY AT THE IMMUNE CELL CHANGES THAT ARE OCCURRING IN EBOLA VIRUS AT THE ORGAN LEVEL. FOR THIS WE CHOSE THE TRANSLOCATOR PROTEIN T IS AN OUTER MITOCHONDRIAL RECEPTOR. SO TYPICALLY USED TO IMAGE NEUROINFLAMMATION. WE KNOW THAT IT IS EXPRESSED EXTRA CRANIALLY AND WE KNEW IT IS EXPRESSED IN CERTAIN BLOOD CELLS BUT WE WEREN'T SURE EXACTLY WHICH IMMUNE CELLS OR BLOOD CELLS ARE THE ONES THAT ARE EXPRESSING. SO WE DID A FLOW CYTOMETRY STUDY WHERE WE FOUND THAT BOTH MOVEMENTS AND REESEES MA CAKES HAVE THE HIGHEST EXPRESSION OF -- RHESUS MACAQUES -- FOLLOWED BY THE NEUTROPHILS AND THE LEAST AMOUNT OF EXPRESSION IN T CELLS AND B CELLS. WE DID SEE SOME EXPRESSION IN THOSE. SO WE THOUGHT WE POTENTIALLY DETECT THE CHANGES THAT ARE OCCURRING AT THE ORGAN LEVEL AND LONGITUDINALLY. THE STUDY DESIGN WAS SIMILAR TO THE ONE DONE BY DR. JOHNSON WHERE THE RHESUS ANIMALS ARE FIRST IMAGED AT BASELINE AND THEN AT MULTIPLE TIME POINTS UNTIL THE TERMINAL TIME POINT. WE HAD A APPROACH SO WE CAN IMAGE ANIMALS OR OBTAIN HISTOLOGY AT DIFFERENT TIME POINTS. VI REAM YUM AND CYTOKINE LEVELS WERE ASSESSED AS WELL AS -- VIREMIA -- WE HAD THE TIM CALL PROGRESSION OF DISEASE. THERE IS NEUTROPHILIA AS WELL AS MONOCYTEO PENIA, LOSS OF PLATELETS AND LOSS OF LYMPHOCYTES WHICH CORRESPOND TO INCREASES IN PLASMA VIRAL LOADS AND THE TISSUE VIRAL LOADS INCREASE BETWEEN DAY 4 AND DAY 6. AND THERE WAS A LOT OF INFLAMMATORY CHANGES OCCURRING ESPECIALLY IN THE LATE STAGE OF THE DISEASE. INTERLEUKIN 10 AND 15MCPT1 AND INTERLEUKIN 18. SO WE LOOKED AT THE IMAGES AND WE CONCENTRATED ON A FEW ORGANS, MAINLY SPLEEN, LUNGS AND BONE MARROW. SO WHEN YOU LOOK AT THE SPLEEN AND THESE IMAGES, THESE ARE IMAGES OVERLAID ON CT SCANS AND THE SPLEEN IS IN GREEN AT THE BASELINE. YOU CAN SEE THAT ON DAY 7 THE SPLEEN INCREASED IN SIZE. BUT THEN THE DISTRIBUTION VOLUME DECREASED. AND THAT DECREASE WAS PROGRESSIVE AROUND THE COURSE OF THE DISEASE CULMINATING NEAR THE TERMINAL ENDPOINTS. THE LUNGS ALSO YOU CAN SEE AT BASELINE THERE IS CERTAIN UPTAKE OF TPA714 AND THAT AGAIN ACTUALLY DECREASED ON THE FOLLOW-UP IMAGES. AND YOU CAN SEE IT HERE IN THIS GRAPH. IT'S DECREASING SIGNIFICANTLY AND BOTH WORKS SIGNIFICANTLY. WHAT BEHAVED DIFFERENTLY IN THIS CASE YOU CAN SEE THAT THE BONE MARROW AND THE SPINE HERE YOU CAN TELL THAT IT'S INCREASING IN SIGNAL AND THE DISTRIBUTION OF VOLUME ALSO INCREASED SIGNIFICANTLY. SO WE WENT BACK AND TRIED TO CORRELATE WITH VARIOUS BLOOD COUNT AND WITH PLASMA VIREMIA. THE SKIN AND LUNG DISTRIBUTION VOLUME VALUES CORRELATED POSITIVELY WITH LYMPHOCYTES AND PLATELETS BECAUSE THEY WERE DECREASING AT THE SAME TIME AS DECREASE IN SPLEEN VOLUME AND ALSO NEGATIVE CORRELATION WITH PLASMA VIRAL LOAD AND NEUTROPHILS AND THE SAME CAN BE SEEN IN THE LUNGS AS WELL. SO WE CONCLUDED THAT BASED ON OUR IMAGING, THERE IS PROBABLY A CERTAIN AMOUNT OF CELLULAR DEPLETION THAT IS HAPPENING IN BOTH ORGANS. IN THE SPLEEN WE THOUGHT THAT THERE IS PROBABLY MONOCYTES AND LYMPHOCYTES THAT ARE BEING DISPATCHED TO THE POLICE OFFICERY AS WELL AS APOPTOSIS THEMSELVES AND IN THE LUNGS WE ASSUME THERE IS APOPTOSIS OF MACROPHAGES BECAUSE WE KNOW THAT MACROPHAGES GET INFECTED WITH THE VIRUS. AND WE THOUGHT THAT THERE WASN'T ENOUGH NEUTROPHILIC ACTIVATION TO COMPENSATE FOR THIS LOSS. THE HISTOLOGY CONFIRMED OUR FINDINGS. ESPECIALLY IN THE SPLEEN WHERE WE FIRST STARTED SEEING LYMPHOIDS AROUND BE DAY 4 AND THEN THAT PROGRESSED TO NECROSIS ON DAY 7. THE IMMUNOFLUORESCENT IMAGING SHOWED US MORE DETAIL PICTURE OF WHAT IS HAPPENING AT THE LEVEL OF DIFFERENT CELL POPULATIONS. SO ON TOP HERE YOU HAVE THE CONTROL ANIMAL AND THIS IS THE INFECTED ANIMAL AND YOU CAN SEE THAT THERE IS DEFINITE DECREASE IN T CELLS. DEFINITE DECREASE IN MONOCYTES WHICH WAS CONSISTENT WITH OUR IMAGING FINDINGS. AND YOU CAN TELL THAT THERE IS MORE LOCALIZING WITH THE MONOCYTES THAN WITH THE LYMPHOCYTES. AT THE SAME TIME, THERE WAS EVIDENCE OF INFECTION SO A EBOLA MATRIX, THE MAJORITY OF THE CELLS ARE COINFECTED AND WERE MONOCYTES. AND THEN THERE WAS A LOT OF APOPTOSIS INVOLVED BOTH LYMPHOCYTES AND MONOCYTES AND ALL THOSE REPORTS WERE CONFIRMED QUANTITATIVELY. OUR IMAGING FINDINGS WERE CONFIRMED BY HISTOLOGY. SUPPORTING THE IDEA THERE IS ELEMENT OF INFECTED APOPTOTIC CELLS AS WELL AS DISPATCH OF VARIOUS CELLS TO THE CIRCULATION. SO IT'S AN ORG-LEVEL DEPLETION RATHER THAN INFLAMMATION. IN THE LUNGS, WE DID SEE SOME NEUTROPHILIA. THERE WAS SOME ATTEMPT BY THE BODY TO FIGHT THE INFECTION. THIS IS INFECTED ANIMAL ON DAY 4. AND YOU CAN STILL SEE SOME MONOCYTES AT THAT LEVEL. AND THEN MACROPHAGES AND THEN IN THE TERMINAL TIME POINT, YOU CAN SEE THERE WAS FURTHER DEPLETION OF THOSE MACROPHAGES AND THE NEUTROPHILIC REACTION DIDN'T REALLY CHANGE MUCH. IN THOSE FINDINGS, CONFIRMED QUANTITATIVELY AND SUPPORTED OUR IMAGING FINDINGS WHERE THERE WAS EVIDENCE OF LOSS OF MACROPHAGES DUE TO INFECTION AND ALTHOUGH THERE WAS INCREASE IN COUNTS IN THE LUNGS, THAT WAS NOT ENOUGH TO COMPENSATE FOR THE LOSS OFAL BOOM MACROPHAGES. AS I MENTIONED EARLIER, THE ONLY ORGAN THAT WENT IN THE OPPOSITE DIRECTION WAS ACTUALLY THE BONE MARROW. AND THE BONE MARROW DISTRIBUTION OF VOLUME CORRELATED WITH THE WBC COUNTS INCREASING AS WELL AS NEUTROPHILS AND VIRAL LOAD AND CORRELATED NEGATIVELY WITH PLATELETS, MONOCYTES AND LYMPHOCYTES. SO WE DIDN'T HAVE HISTOLOGY FROM THE BONE MARROW BUT WE BELIEVE THAT BASED ON ALL THESE CORRELATIONS AND CORRELATIONS WITH PLASMA CYTOKINES, THESE CHANGES IN THE BONE MARROW PROBABLY REFLECT EXPRESSION IN HEMATOPOIETIC STEM CELLS AND WE BELIEVE THOSE ARE ACTIVATED BIOPOSING MECHANISMS. FIRST ONE IS SUPPORT FOR LIKEO. [ SIGH ] TOSIS -- LEUKOCYTOSIS AND THE SECOND ONE IS COMPENSATION FOR DEPLETION OF MONOCYTES AND PLATELETS. AN ELEMENT OF INFLAMMATION EFFECT ON HEMAPOIESIS THAT IS INCREASING THE ACT OR INCREASED EXPRESSION OF BONE MARROW. WE BELIEVE THAT BONE MARROW DISTRIBUTION OF VOLUMES COULD POTENTIALLY PROVIDE PROGNOSTIC INFORMATION ON THE RECOVERY ATTEMPTS AND MIGHT INDICATE WHAT THE ODDS FOR SURVIVAL ARE. SO IN CONCLUSION FOR THIS PART, USING THE PET IMAGING, WE WERE ABLE TO VISUALIZE ORGAN LEVEL EBOLA PATHOGENESIS. [ READING ] WE BELIEVE HAVING ESTABLISHED BIOMARKER OF DISEASE PROGRESSION, IT CAN USE TO ASSESS THE EFFECTIVENESS OF VARIOUS INTERVENTIONS LIKE TREATMENTS AND VACCINES. WE ALSO THINK THIS IS A SIMILAR APPROACH WITH THE OTHER LIGANDS COULD BE USED TO UNDERSTAND THE INTERPLAY BETWEEN THE PATHOGEN AND THE HOST IMMUNE REACTIONS IN OTHER SYSTEM INFECTIONS POSSIBLY LIKE COVID-19. SO THE SECOND PART OF MY STUDY IS ABOUT USING MOLECULAR IMAGING MODALITIES TO BETTER UNDERSTAND MANIFESTATIONS OF EBOLA VIRUS DISEASE. AFTER THE EBOLA EPIDEMIC WAS CONTROLLED, WE STARTED SEEING A NEW SYMPTOM IN SURVIVORS AND THIS WAS NAMED POST EBOLA DISEASE SYNDROME. IF YOU LOOK AT THE SYMPTOMS, THEY ARE SIMILAR TO WHAT WE ARE SEEING NOWADAYS IN PATIENTS WITH COVID. SO EXTREME FATIGUE, DIFFICULTY SLEEPING, CHRONIC HEADACHES, MEMORY LOSS, SOME PATIENTS HAD MOOD DISORDERS AND A FEW COMPLAINED OF VISUAL AND AUDITORY PROBLEMS. SO THE QUESTION WAS, WHAT IS HAPPENING IN THE WHY. NS OF THOSE PATIENTS AND -- IN THE CNS OF THOSE PATIENTS AND WHY ARE THEY BEING LEFT WITH ABNORMALITIES OR NEUROLOGIC PROBLEMS? AND AT THAT POINT, WE DIDN'T REALLY KNOW WHAT WAS HAPPENING IN THE BRAIN. THE THEORY WAS THAT THERE IS AN ELEMENT OF NEUROINFLAMMATION SINCE THIS IS INFECTIOUS PROCESS AND THERE WERE PREVIOUS STUDIES WHERE RHESUS MACAQUE EBOLA SURVIVORS SHOWED INIA FLORIDAIMATION. WE ALSO -- INFLAMMATION. AND THE VIRUS WAS DETECTED IN THE CSF IN MULTIPLE REPORTED CASES. WE ALSO THOUGHT THERE IS AN ELEMENT OF BLOOD-BRAIN BARRIER DISRUPTION. MOSTLY BASED ON THE FACT THAT THERE IS ENDOTHELIAL INVOLVEMENT IN THE BRAIN THAT HAS BEEN SHOWN PREVIOUSLY SIMILAR TO WHAT HAPPENS IN THE PERIPHERY. SO THAT COULD POTENTIALLY BE CAUSING BLOOD-BRAIN BARRIER DISRUPTION AS WELL AS SYSTEMIC INFLAMMATION WHICH IS KNOWN TO CAUSE A CERTAIN AMOUNT OF DISRUPTION IN THE BLOOD BRAIN BARRIER. SO WE DID A VERY SIMILAR STUDY TO THE PREVIOUS ONE WHERE WE HAD RHESUS MACAQUE ANIMALS IMAGED AT BASELINE INFECTED WITH EBOLA AND ASSESSED LONGITUDINALLY UNTIL TERMINAL TIME POINTS. WE HAD PATHOLOGY FROM DAY 4 AS WELL AS TERMINAL TIME POINTS. WE HAD VIREMIA AND CYTOKINE LEVELS IN THE PLASMA AS WELL AS IN THE GROUP THAT UNDERWENT MRI AND HAD CSF VALUES AS WELL. AND THE IMAGING INCLUDED EASTERN FFDG PET TO TRACK CHANGES IN GLUCOSE METABOLISM TO REFLECT NEURAL FUNCTION AND NEUROINFLAMMATORY CHANGES AND ALSO PERFORMED MRI IMAGING WITH T1 MAPPING TO SEE IF THERE ARE ANY STRUCTURAL ABNORMALITIES AND ASSESS FOR BLOOD-BRAIN BARRIER INTEGRITY. AND THERE IS ALSO A VERY INTERESTING, WHEN WE LOOKED AT THE BRAIN AS A WHOLE, THERE WERE NOTHING CHANGES IN THE WHOLE BRAIN BETWEEN BASELINE AND LAST TIME POINTS OR CROSS TIME. BUT THEN WHEN WE LOOKED AT THE RELATIVE UPDATE IN VARIOUS REGIONS WE NOTICED REDISTRIBUTION AND CERTAIN REGIONS SHOWED INCREASED UPTAKE OVER TIME AND MOST OF THE THAT WERE STATISTICALLY SIGNIFICANT INCLUDED PONDS, MID BRAIN AND THALAMUS. YOU CAN SEE IT HERE IN THE MID RAIN, INCREASED FDG UPTAKE AND IN THE CEREBELLUM. SO WE WENT BACK AND TRIED TO CORRELATE WITH CYTOKINE VALUES AND AS EXPECTED, BECAUSE WE KNEW THE PATIENTS WERE HAVING A LOT OF INFLAMMATORY CHANGES, THERE WERE DEFINITE CORRELATIONS BETWEEN THE INCREASING UPTAKE AND VALUES OF CYTOKINES IN THE PERIPHERY ESPECIALLY WITH INTERLEUKIN 8 AND 6. AND THEY CORRELATED WITH INTERLEUKIN 10 AND PLASMA VIRAL LOAD. SO, WE BELIEVE THAT THIS INCREASED UPTAKE AT LEAST PARTIALLY IS DUE TO THESE NEW INFLAMMATORY CHANGES AND ON HISTOLOGY WE SAW THERE WERE ENLARGED CELL BODIES OF THE MICROGLIA WHICH WERE CONSISTENT WITH EARLY DNA ACTIVATION. WE BELIEVE WE DIDN'T SEE THE FULL ACTIVATION THAT YOU USUALLY SEE IN SLIGHTLY MORE CHRONIC DISEASES MAINLY BECAUSE THE ANIMALS DIED QUICKLY SO THERE WASN'T ENOUGH TIME TO DEVELOP THOSE PHENOTYPIC CHANGES OF NEUROINFLAMMATION. BUT WE WERE ALSO -- WE NOTICED THAT THERE WERE CERTAIN GROUPS OF NEURONS THAT WERE STAINING. THIS IS A CONTROL ANIMAL AND THIS IS EBOLA VIRUS INFECTED ANIMAL AND SOME OF THOSE NEURONS WHICH STAINED POSITIVELY ALSO STAINED FOR VP40. AND ALMOST ALL WERE SHOWING APOPTOTIC CHANGES T SEEMED LIKE CLUSTERS OF NEURONS ARE GETTING INFECTED AND THEY WERE ACTUALLY UNDERGOING APOPTOSIS. AND THAT'S NOT VERY UNEXPECTED BECAUSE WE KNOW THAT THE RECEST OB IS ACTUALLY EXPRESSED ON NEURONS AND THAT IS WHAT THE EBOLA VIRUS USES TO GAIN ENTRY INTO CELLS. BUT WE ALSO WERE SURPRISED BECAUSE WE DID STAIN FOR GLUCOSE TRANSPORTERS AND FOUND THAT THERE WAS AN UPREGULATION OF GLUCOSE TRANSPORTEE TYPE III, THE NEURONAL TYPE OF GLUCOTRANSPORTER, ESPECIALLY IN THE APOPTOTIC CELLS AS YOU CAN SEE HERE. AND THIS IS NOT SOMETHING THAT HASN'T BEEN DESCRIBED BEFORE. IT HAS BEEN DESCRIBED IN A FEW OTHER ENTITIES LIKE BRAIN DAMAGE AND OTHER EXAMPLES OF GLUTAMATE EXCITOTOXICITY. WE BELIEVE THAT WHAT HAPPENED HERE IS ALSO AN UPREGULATION IN THOSE NEURONS AND THE APOPTOTIC NEURONS WHICH COULD ACCOUNT FOR THE INCREASE IN UPTAKE WE WERE SEEING ON FDG PET AND MOST OF THOSE NEURONS WERE SEEN IN THE BRAINSTEM STRUCTURES. SO THE SECOND PART WAS TO EVALUATE THE BLOOD-BRAIN BARRIER AND FOR THIS WE PERFORMED -- ESSENTIALLY WHAT WE ARE DOING TO SIMPLIFY IT -- [ READING ] SO IF YOU HAVE ACCUMULATION OF FLUID THE T1 VALUES ARE EXPECTED TO INCREASE. SIMILARLY, IF YOU ADMINISTER CONTRAST DUE TO THE INHERENT EFFECTS OF GADOLINIUM, YOU WILL END UP WITH FURTHER FACILITATION OF LONGITUDINAL RELAXATION WHICH MEANS THE T1 VALUE WILL DROP MORE SIGNIFICANTLY AFTER CONTRAST IF YOU HAVE CONTRAST INTO THE EXTRACELLULAR SPACE. SO WE DID THOSE EXPERIMENTS ON A SEPARATE GROUP OF ANIMALS AND IF YOU LOOK HERE AT THIS ANIMAL, THIS IS ONE OF THE ONES WHO WERE IMAGED AT DAY 5 POST KNOCKALATION AND THESE ARE T1 MAPS. AND YOU CAN SEE EVEN VISUALLY THESE ANIMALS SLIGHT INCREASE IN T1 VALUES OVER TIME AND ALTHOUGH WE SAW THIS ALMOST THROUGHOUT THE WHOLE BRAIN, THE REGIONS WITH MOST SIGNIFICANT WERE THE WHITE MATTER. AND AGAIN AFTER ADMINISTRATION, YOU CAN SEE THESE MAPS WERE ACTUALLY MORE DECREASED IN T1 VALUES AFTER KNOCKALATION COMPARED TO BASELINE. THIS SHORTENING OF T1 VALUES WAS SEEN ALSO ACROSS THE BRAIN BUT WAS MOSTLY IN THE THALAMUS. BOTH VALUES T1 AND T1 SHORTENING CORRELATED WITH VIREMIA AND CSF CYSTOKINES. IMMUNOFLOURESENCE SUPPORTED OUR FINDINGS. YOU CAN SEE THIS IS ANIMAL THAT WAS SACRIFICED ON DAY 4. THIS WAS SACRIFICED ON DAY 6 AND YOU CAN SEE THAT THERE IS PROGRESSION OF ALBUMIN LEAKAGE HERE STAINED IN GREEN. SO THERE WAS DEFINITE LEAKAGE OF ALBUMIN IN THOSE ANIMALS WHICH WORSENED OVER TIME AND THAT CONFIRMED THERE IS A ELEMENT OF BLOOD-BRAIN BARRIER DISRUPTION. AND THE PROBLEM WITH THE DISRUPTION IS IT EXPOSINGS NEURONS FROM ALL KINDS OF TOXINS FROM THE PERIPHERY AND ONE OF THOSE -- ONE OF THE PRODUCKS THAT COULD CAUSE NEURONAL DEATH IS ALBUMIN. SO THAT COULD POTENTIALLY EXPLAIN WHY SOME OF OUR NEURONS WERE STAINING FOR ALBUMIN AND WERE ALSO BEING APOPTOTIC. SO IN CONCLUSION WHAT I WAS TRYING TO SHOW HERE IS THAT IMAGING CAN BE INSTRUMENTAL IN HIGH CONTAINMENT VIRAL INFECTIONS BECAUSE THEY COULD HELP ELUCIDATE DISEASE PATHOPHYSIOLOGY AND BECAUSE YOU COULD DEVELOP NON-INVASIVE BIOMARKERS TO ASSESS EFFECTIVENESS OF PREVENTIVE AND THERAPEUTIC INTERVENTIONS. THE GOOD THING ABOUT IT IS THAT THE SAME APPROACHES AS WELL AS OTHER MOLECULAR TECHNIQUES CAN ALSO BE APPLIED TO OTHER INFECTIOUS AGENTS AS WELL AS CHEMICAL AND OTHER BIOLOGICAL WARFARE. AND THAT WILL PROBABLY IMPROVE OUR UNDERSTANDING OF THE PATHOPHYSIOLOGY OF DISEASE ASSOCIATED WITH THOSE INFECTIONS AND IMPROVE ABILITY TO TEST VACCINES AND TREATMENT. WITH THAT ISLAND LIKE TO THANK THE IRF TEAM. IT'S A HUGE AMOUNT OF WORK AND I REALLY GRATEFUL FOR THIS COLLABORATION BECAUSE NONE OF THIS WOULD HAVE BEEN DONE WITHOUT THEM. ALSO THE CLINICAL CENTER, OBVIOUSLY MY GROUP AND THE PHYSICIAN AS WELL AS THE CHEMISTRY AND SYNTHESIS CENTER AND NINDS AND NIAID. THANK YOU FOR YOUR ATTENTION. >> THANK YOU. THAT WAS SUCH A WONDERFUL TALK. OUR LAST SPEAKER IS A RESEARCH FELLOW AT JOHNS HOPKINS. HE WILL SHARE HIS WORK ON PET IMAGING IN SARS-COV-2. >> THANK YOU. CAN YOU HEAR ME AND SEE THE SCREEN? >> WE CAN HEAR YOU AND NEED TO SWAP YOUR SCREEN. PERFECT. >> SO THANK YOU FOR HAVING ME HERE. I'M REALLY EXCITED TO PRESENT OUR WORK AND SARS-COV-2 AND I KNOW IT HAS BEEN A LONG DAY BUT IT'S BEEN REALLY EXCITING TO HEAR ABOUT ALL THE WORK THAT HAS BEEN DONE IN BACTERIAL IMAGING AND IMMUNOIMAGING AND EXCITING MODELS WE HAVE BEEN TRYING AND ALSO PUSHING ALL OF THIS TECHNOLOGY INTO THE CLINIC. I'M GOING TO TRY TO BE BRIEF. SO I KNOW YOU MIGHT BE TIRED SO I'M GOING TO TRY TO GO THROUGH THIS WHOLE RESULTS AND IDEAS QUITE FAST. SO, BASICALLY WE HAVE GONE THROUGH THIS BUT ONE OF THE THINGS THAT WE START TO SEE WHEN THE PANDEMIC STARTED IS THAT WE STARTED TO GET SOME RESULTS AND SOME IDEA WHAT WAS THE IMMUNE LANDSCAPE ESPECIALLY IN ONE COMPARTMENT OF PLASMA. WITH TIME WE START GETTING MORE IDEA WHAT WAS HAPPENING IN OTHER COMPARTMENTS AS WELL AS IN THE COMPARTMENTS AS WELL AS IN THE TISSUE. AND WE ARE STARTING TO LEARN THAT THERE IS SPECIFIC SALESCOULD BE UP OR DOWN REGULATED THROUGHOUT THE PROCESS OF INFECTION. WE WERE THAT IN THE PLASMA COMPARTMENTS CELLS ARE LIKE LYMPHOCYTES AND COULD DECREASE THROUGHOUT THE INFECTION BUT IN THE TISSUE FOR EXAMPLE WE ALSO SEE THAT THE LYMPHOCYTE, CD4 AND CD8 AND POSSIBLY INCREASING AS WELL AS MYELOID CELLS SPECIFICALLY MONOCYTES AND MACROPHAGES. AND NOW THAT WE ALSO SEE SPECIFIC TISSUES AND ESPECIALLY BY IMAGING CYTOMETRY, WE HAVE BEEN ABLE TO SEE THESE CELLS, ESPECIALLY THE MYELOID AND MACROPHAGE CELLS AND MONOCYTES ARE QUITE IMPORTANT IMMUNE CELLS RELATED TO THE ACTIVATION AND RESPONSE TO THIS. SO ALSO ANOTHER THING WE HAVE BEEN LEARNING IS THAT IN GENERAL WORLDWIDE THERE HAS BEEN A DISCREPANCY BETWEEN WHAT IS HAPPENING BETWEEN FEMALES AND MALES AND WE ARE SEEING THAT ESPECIALLY IN MALES SUFFERING FROM -- MORE FATAL AND MORTALITIES INCREASED BETWEEN THEM AND THERE ARE MULTIPLE THEORIES BEHIND WHAT COULD BE HAPPENING AND WHY THE DISCREPANCY ARE HAPPENING RIGHT NOW. SO THEY ARE ALSO IN BETWEEN THOSE WE ARE SEEING IT COULD BE RELATED TO THE VIRUS ENTRY, RELATED TO THE IMMUNE RESPONSE, ALSO TO THE LACK OF IMMUNE RESPONSE AND HUMAN RESPONSE AND COMPARISONS BETWEEN FEMALES AND MALES ESPECIALLY FROM WHAT WE HAVE BEEN SEEING IN OTHER VIRAL INFECTIONS. RELATED TO THIS THERE IS ALSO A THEORY BEHIND THERE COULD BE A HUMAN RESPONSE ASSOCIATED WITH HORMONAL RESPONSES. SOME OF THE CLINICAL TRIALS ASSOCIATED WITH THIS ARE LISTED HERE. TWO OF THOSE WE ARE TRYING TO LOOK AT SPECIFIC CONCENTRATIONS OF ESTROGEN AND HOW THEY CAN CHANGE POTENTIALLY THE MORBIDITY AND MORTALITY OF MALES THROUGHOUT THE DISEASE. AND I THINK THAT IT IS NOT FULLY NECESSARY TO EXPLAIN DETAILS BECAUSE I THINK WE HAVE BEEN HEARING A LOT OF WONDERFUL TALKS ABOUT THE IMPORTANCE OF MOLECULAR IMAGING, ESPECIALLY TO GO THROUGH MOLECULAR IMAGING TO FIND PATHOGENS AS WELL AS TO UNDERSTAND OTHER MICRO-ENVIRONMENTAL VARIABLES SUCH AS INFLAMMATION AND HYPOXIA. RIGHT NOW WHAT I'M GOING TO SHOW YOU IS ONE OF THE TRACERS THAT CAN BE USED TO DETECT SPECIFICALLY SOME OF THE IMMUNE CELLS AND TARGET TO UNDERSTAND THE PATHOPHYSIOLOGY OF SARS. IN THIS CASE, WE ARE LOOKING TO DPA13 AND IS USED AS PART OF THE TRACER TO LOOK AT MACROPHAGES OR MONOCYTE CELLS IN SEVERAL MODELS OF PUMINARY ADT. AND WE HAVE SEEN THEY QUITE ACCUMULATE IN CELLS WITH POSITIVE CELLS AS WELL AS THAT ARE CENTERED WITH MACROPHAGES WITHIN THE TISSUE AND ALSO CORRELATES WITH WHAT IS HAPPENING THROUGHOUT TREATMENT WITH FOR EXAMPLE IN THE CASE OF PULMONARY ADT CORRELATED WITH THE BACTERIAL BURDEN. AND ALSO IT'S BEEN ALREADY BEEN USED IN HUMANS, HEALTHY VOLUNTEERS AND SEEMS TO BE FAVORABLE. WE COULD BE POTENTIALLY USING THIS AS A TRANSLATABLE TOOL IN THE FUTURE. IN GENERAL TERMS WE DID QUITE SIMPLE EXPERIMENT WHERE WE WANTED TO SEE WHAT WAS HAPPENING WITH THE DPA AND THE DISTRIBUTION OF DPA IN HAMPSTERS. AND BASICALLY WE USED DP7, THE SEVERITY IS HIGHER AT DAY 7 AND THEN START DECREASING THROUGH ALL-TIME. THE HAMPSTER IS CONSIDERED A MODEL WITH MILD DISEASE BUT THEY ACTUALLY SHOW SOME INTERESTING FEATURES IN PULMONARY DISEASE AS IN THE PATHOPHYSIOLOGY AND IMMUNE RESPONSE AND THEN WE INTRODUCED ANIMALS AT DAY 7 AND AFTER THAT, WE COLLECTED TISSUE TO PERFORM SOME READ OUTS ON THE TISSUE FOR HISTOPATHOLOGY, FLOW CYTOMETRY AND IMMUNOHISTOCHEMISTRY. WE FOUND AND REPORTED BEFORE THAT WE ARE SEE SOMETHING FISSURES THAT CAN BE CORRELATED TO THE HUMANS, SOME OF THEM WITH GROUND GLASS CAPACITIES AND SOME OF THEM HAMPSTERS SEEN IN HUMAN PATIENCE AND ALSO REPORTED BUT WE ARE SEEING HERE MORE AND ALSO SOME AREAS OF CONSOLIDATION IN THE RIGHT LUNG AND IN THE UPPER LOBES IN THIS CASE IN HAMPSTERS. SO AS WE HAVE BEEN DISCUSSING AS WELL, WE WANTED TO FIND A WAY TO QUANTIFY THE DISEASE IN THE LUNGS AND WE ARE ABLE TO DEVELOP A AUTOMATED TOOL TO TRY TO FIND THE AREAS OF PNEUMONIA OR HYPERTENSE AREAS THAT WERE ASSOCIATED WITH THE DISEASE IN A WAY WE CAN LOOK AT THE THREE-DIMENSIONAL AND THE WHOLE LUNG BY CREATING A WHOLE LUNG AND THEN SPECIFIC AREA FOR THE PNEUMONIC AREAS AND THROUGHOUT DISEASE. AND THEN WE QUANTIFY THIS DISEASE, ESPECIALLY WE WANTED TO SEE NOW THAT WE KNEW THE DISEASE WAS HIGH, WE WANTED TO SEE WHAT WAS DIFFERENT BETWEEN MALES AND FEMALES. INTERESTINGLY WE FOUND THAT DESPITE THE FA ACCOUNT THAT THE WHOLE VOLUME AND THE ROI FOR THE WHOLE VOLUME WAS SIMILAR BETWEEN THEM, THE VOLUME OF CONSOLIDATION AND THE PERSONAL CONSOLIDATION WAS SIGNIFICANTLY HIGHER IN MALES THAN IN FEMALES. WHEN WE LOOK AT THE DPA SIGNAL THAT WAS ASSOCIATED TRYING TO LOOK AT WHAT WAS HATCHING WITH THE MYELOID CELLS CLOSER TO THE MONOCYTE AND MACROPHAGE RESPONSE, WE SEE THAT THERE WAS A LOCALIZED ACCUMULATION OF THE SIGNAL ON THE AREAS OF GROUND GLASS CAPACITIES AND PNEUMONIA CONSOLIDATION AND WHEN WE SEGREGATED ANALYSIS FOR MALES AND FEMALES FOR HIGHER ACTIVITY OR UPTAKE IN THE MALES COMPARED TO FEMALES. SO ALSO WHEN WE LOOK AT THE DISTRIBUTION FOR THE HAMPSTERS WE SAW SIMILAR DISTRIBUTION IN LITERATURE FOR THIS TRACER WITH LOW PENETRATION OF THE BRAIN AND THE GI TRACT SITUATION ALSO AND SIMULATION ON THE ADIPOSE TISSUE AND AREAS WITH THE MACROPHAGES. SO WE ALSO TRY TO VALIDATE SEEING ESPECIALLY FOR THE LOCALIZATION OF ACTIVITY TO SEE WHAT WAS HAPPENING EXACTLY ON THE LUNGS OF THE HAMPSTER AND ALSO BEING ABLE TO SEE SOME UPTAKE IN LOCALIZED AREAS OF DISEASE DAY 7 POST-INFECTION. FURTHER MORE WE TRIED TO LOOK AT WHAT WAS HAPPENING BELOW THE TISSUE ALL THE TECHNIQUES HAVE SPECIFIC LIMITATION. THIS CASE WE SEE THE ACTIVITY OR THE EXPRESSION ON THE INFILTRATION OF IBA HASSATIVE CELLS WITHIN THE TISSUE. AND BY FLOW CYTOMETRY, WE DON'T HAVE -- RIGHT NOW WE HAVE A LOT OF REAGENTS FOR FLOW CYTOMETRY. WE LOOK AT GRANULARITY AS WELL AS SIZE WHERE WE ARE ABLE TO DIFFERENTIATE SPECIFICALLY BETWEEN LYMPHOCYTE AND MAILOIDS AND THE C11B POSITIVE CELLS AND WE FOUND THAT WHILE THE CD11 POSITIVE CELLS WERE HIGHER AS WELL AS THE MYELOID IN GENERAL IN MALES THE LYMPHOCYTES ON THE OTHER HAND IN HUMANS WAS HIGHER WITHIN THE TISSUE OF THE HAMPSTERS AT DAY 7. SO INTERESTINGLY WE WERE LOOKING FURTHER MORE INTO THIS MODEL AND TRYING TO SEE IF WE WERE ABLE TO, APART FROM THE IMMUNE RESPONSE, IT HAS BEEN ASSOCIATED WITH RESPONSE IN HAMPSTER FEMALES AND WE WERE ALSO SEEING THAT COMPARING BETWEEN PLASMA LUNGS EARLY ANTIBODIES AS IGM. THE TITERS OF IGM WERE HIGHER AS AN AREA OF RESPONSE IN FEMALES AT DAY 7 IN LUNGS COMPARED TO PLASMA. IT WAS ALSO HIGHER IN PLASMA ESPECIALLY AT DAY 21. AND ALSO INTERESTING SHOWING ENTRAPPANCY BETWEEN SEX IN THIS MODEL. SO PART OF THE QUESTION WAS, SO WHAT WE COULD DO TO UNDERSTAND POTENTIALLY THE FACTOR WHO COULD BE ASSOCIATED WITH THIS OUTCOME. SO WE TRIED TO LOOK TO THE EFFECT IN THE DISEASE AND PATHOLOGY WHEN WE USED ESTRADIOL, E2 AT DAY 7 TO SEE IF THE MORBIDITY WILL DECREASE IN MALES. SO WE DID A SIMPLE EXPERIMENT WHERE WE TOOK A PLACEBO MALE AND THEN COMPARED THEM WITH MALES WHO WERE INOCULATED THAT WAS GOING TO HAVE RELEASE OF ESTROGEN UNTIL DAY 7 AT THE SAME TIME THEY START THE INFECTION. AND THEN WE LOOK AT THE WEIGHT AND THE CT SCORE, THE CONCENTRATION OF PLASMA SO HISTOPATHOLOGY. INTERESTINGLY, WE SAW THAT WHILE THE CONCENTRATION OF ESTROGEN WAS SIMILAR TO WHAT HAS BEEN REPORTED AS CYTO-- HAMPSTERS, THE WEIGHT WAS NOT SIGNIFICANTLY DIFFERENT BETWEEN THE GROUPS AND FURTHERMORE THERE WERE NO DIFFERENCE IN THE CT SCORE BETWEEN THE PLACEBO MALES AND THE MALES WITH ESTROGEN. AND IF WE LOOK AT THE CTs, WE ALSO SEE THERE IS OBVIOUS&VISIBLE DISEASE IN BOTH GROUPS WITH SOME AREAS OF PNEUMONIA AS WELL AS LUNG CAPACITIES FOR BOTH GROUPS AND ALL THE ANIMALS, 13 ANIMAL PER GROUP. SO WE FOUND THIS REALLY INTERESTING. WE TOOK THE SECTIONS, LUNGS, AND DID SOME TIME TO UNDERSTAND WHAT WAS THE CELLS INFILTRATING AND THE CHARACTERISTICS OF THE DISEASE WAS SIMILAR TO BETWEEN GROUPS, WE FIND SIMILAR -- MULTINUCLEATED CELLS REPORTED BEFORE AND WERE SIMILAR TO WHAT IS BEING OR HAS BEEN SHOWN FOR MALES AND THE DISEASE WAS ALSO SIMILAR IN HISTOLOGY. IF WE LOOK ALSO AT SOME IBA1 CELLS, WE STILL SEE A LOT OF CELLS WITHIN THE LUNG AND NOT A HUGE DIFFERENCE BETWEEN GROUPS WHICH WAS AN INTERESTING FINDING AS WELL. SO I GUESS WITH THIS, I WOULD LIKE TO CONCLUDE THAT WHAT WE WERE SEEING IS THAT MALES GENERALLY IN THIS MODEL, WE ARE SEEING THAT MALES ARE EXPERIENCING GREAT MORBIDITY THAN FEMALES AND WE WERE NOT ABLE TO SEE REVERSE OF THIS DISEASE WITH ESTROGEN. AT LEAST AT DAY 7. THE FEMALES WERE DEVELOPING GREATER ANTIBODY RESPONSE IN IGM. THERE IS MORE DATA WITH I Q.E.W. AND NEUTRALIZING ANTIBODIES. AND THE DISTRIBUTION OF DPA WAS SIMILAR TO WHAT WE WERE SEEING BEFORE AND ALSO THE ACTIVITY OF THE UPTAKE OF DPA WAS LOCALIZED IN THE AREAS OF INFECTION AND ALSO WHEN WE SURVEYED ANALYSIS IT WAS HIGHER ACTIVITY UPTAKE OF MALES COMPARED TO FEMALES. AND IN FLOW CYTOMETRY, WE WILL NEED TO ANALYZE THIS AND SEEING THAT MYELOID 11B POSITIVE WERE HIGHER COMPARED TO MALES AND FEMALES AND CONVERSELY, FEMALES HAD HIGHER LYMPHOCYTE INFILTRATION IN THE LUNGS. AND WITH THIS, I JUST WANT TO THANK MY MENTORS AND ALSO ALL MY GROUP WHO HAS BEEN WORKING EXTREMELY IN THE HARD IN THE LAB AND TRYING TO WORK WITH SARS AND TRYING TO LEARN AND TRYING TO FORWARD THINGS AND A HUGE EFFORT FROM ANDREW'S LAB, JOSEPH'S LAB AND KATHRYN WHO HAS BEEN THE ONE. AND OF COURSE THANK YOU FOR THE FUNDING AND WITH THIS, I THINK I WILL HAVE TO -- WE ARE READY FOR QUESTIONS. THANK YOU VERY MUCH. >> THANK YOU VERY MUCH CAMILLO AND TO ALL OF OUR SPEAKERS FROM THIS SESSION. I NOW INVITE YOU ALL TO TURN YOUR CAMERAS BACK ON SO WE CAN BEGIN OUR DISCUSSION. IN THE INTEREST OF TIME, WE WILL ONLY BE TAKING ONE QUESTION PER SPEAKER. SO I'M GOING TO START WITH A QUESTION FOR DR. HENRY VANBROCKLIN. SEVERAL MONOCLONAL ANTIBODIES HAVE VARIABLE LOCALIZATION. THE MECHANISM IS NOT COMPLETELY CLEAR AND PROBABLY NOT RELATED TO SECRETION. HOW CAN YOU BE SURE THAT UPTAKE IS NOT NON SPECIFIC RATHER THAN TARGETING YOUR ANTIGEN? >> SO THAT'S A GREAT QUESTION. AND IT'S ONE THAT WE HAVEN'T THOUGHT HOW WE WOULD TACKLE. I MEAN I WANT TO SAY THIS AND I DIDN'T SAY IT DURING THE PRESENTATION, THAT THE PATIENTS THAT WE HAVE IMAGED ARE INCREDIBLY COOPERATIVE. THE GROUP OF PATIENTS THEY ARE HUGE ADVOCATES FOR PARTICIPATING IN CLINICAL TRIALS AND STUDIES. AND AS I NOTED THERE ARE SEVERAL OF THEM SUBJECTED THEMSELVES TO BIOPSY OF LYMPH NODES SO THAT WE COULD GET THE P24 DATA. I DON'T KNOW -- IF ANY WILL -- AND WHAT OTHER BIOPSIES THAT WE MIGHT HAVE AVAILABLE RELATIVE TO THE BOWEL. I HAVE TO TALK TO TIM ABOUT THAT THAT. BUT IF THERE ARE NONE, WE'LL HAVE TO COME UP WITH SOME OTHER WAYS TO DETERMINE WHAT SIGNAL THERE IS REAL AND WHAT IS MAYBE NON SPECIFIC BINDING. BUT I AGREE THAT WE DID SEE SOME UPTABLING IN THE BOWEL IN THE NORMAL HEALTHY CONTROL VOLUNTEERS. AND SO THERE IS PROBABLY SOME NON SPECIFIC NATURE TO THAT UPTAKE. >> THANK YOU VERY MUCH FOR YOUR ANSWER. QUESTION FOR DIMA HAMMOUD. IS THE BLOOD-BRAIN BARRIER DISRUPTION LOCALIZED WITH SPECIFIC AREA OF THE BRAIN? REALLY INTERESTING DATA. >> THANK YOU. SO ACTUALLY WE SEE ALMOST ACROSS THE WHOLE BRAIN BUT IT WAS MORE PATCHY AND I THINK THERE WAS A LITTLE BIT MORE LIKE THE AREAS THAT WERE STATISTICALLY SIGNIFICANT FOR MOSTLY THE WHITE MATTER CONTAINED IN THE THALAMUS. WE DON'T KNOW WHAT THAT MEANS. WHY WOULD THAT BLOOD-BRAIN BARRIER BE SPECIFIC IN THOSE LOCATIONS? IT COULD BE ONE OF THOSE WEIRD VIRAL PREDICTIONS OR COMPLETELY BY CHANCE. BUT THOSE WERE THE THREE MAIN REGIONS THAT WERE INVOLVED. >> THANK YOU. QUESTION FOR CAMILLO. USING FLOW, DID YOU DO AN INTRACELLULAR PROTOCOL TO LOOK AT LEVELS AND INVESTIGATE WHAT CELLS ARE EXPRESSING TSPO DURING SARS-COV-2 EXPRESSION AS WELL AS A FUNCTION OF SEX? >> GREAT QUESTION. SO WE DIDN'T LOOK AT TSPO. WE DID LOOK AT THE CELLULAR -- THE CELLS THAT WERE DPA713 POSITIVE THAT WERE ALSO 11B POSITIVE. WE FOUND SIMILAR FINDINGS TO WHAT IS BEING REPORTED FOR OUR MODELS IN MICE AND SO BUT WE DIDN'T LOOK AT SPECIFICALLY FOR TSPO. WE KNOW THAT BASED ON OUR PROTOCOLS AND BASED ON PREVIOUS DATA, 24 HOURS PLUS INJECTION COLOCALIZES THE -- WE KIND OF VEERED FROMMED THAT, THAT COULD BE ASSOCIATED WITH THE SAME CELLS THAT ARE TSPO POSITIVE. AND WE HAVEN'T DONE THIS DATA ANALYSIS BETWEEN MALES AND FEMALES FOR THAT. >> THANK YOU. AND ONE LAST QUESTION JUST IN INTEREST OF TIME TO DR. IAN CROZIER. COULD YOU COMMENT -- SO THERE WAS A LOT OF -- NOT A LOT BUT SOME VERY INTERESTING OVERLAP AND SIMILARITIES BETWEEN THE MONKEY AND THE HAMPSTER MOD ELSE FOR THE LAST TWO TALKS. SO COULD YOU COMMENT WHAT YOU THINK WOULD BE ADVANTAGES OF USING THE MONKEY MODEL OVER THE HAMPSTER? YOU'RE MUTED. I'M SORRY. >> CERTAINLY WE DO A -- MOST OF OUR BANDWIDTH HEADS BEEN OCCUPIED BY HAMPSTER WORK IN THE LAST YEAR SO CERTAINLY THE HAMPSTER APPEARS TO BE MORE USEFUL MODEL OF SEVERE DISEASE. AND WE HAVE BEEN INTERESTED IN THE HIGHER RESOLUTION LOOKS AT THE LUNG AND QUANTIFYING THAT. AS WELL AS I SUPPOSE, THE DIRTH. IT'S GROWING BUT THE DIRTH OF TOOLS WITH WHICH TO INTERROGATE THE HAMP STIR. I THINK THEY ARE SERVING INCREDIBLE PURPOSES AND THE HAMPSTER IS SERVING A MORE -- IN SOME REGARDS, WE HAVE BEEN DOING A LOT OF EVALUATIONS, THERAPEUTICS IN THE HAMPSTER MODEL AND LESS OF THE INTERROGATION OF THE IMAGING PHENOTYPE ALTHOUGH WE HAVE DONE SOME OF THAT SO I WAS REALLY INTERESTED IN WHAT CAMILLO PRESENTED. >> THANK YOU. SO THERE IS NO MORE TIME FOR QUESTIONS. THANK YOU TO ALL OF OUR SPEAKERS FOR THE LAST SESSION. I REALLY ENJOYED IT. TERRIFIC PRESENTATIONS. NOW TO WRAP UP OUR SYMPOSIUM, I INVITE DR. JANE TO GIVE US FINAL REMARKS. >> THANK YOU. DEMA DO YOU WANT TO SAY SOMETHING? I CAN JUST START BY SAYING. >> I WILL JUST SAY A COUPLE OF SENSES AT THE END. >> SO SHE ASKED ME TO TALK ABOUT -- A WRAP UP SESSION AT THE END SO I JUST WANT TO TELL EVERYBODY AGAIN, I'M JUST GOING TO SHARE MY SLIDES. WHAT I REALLY WANTED TO REMIND EVERYBODY IS THAT WE ARE TALKING TODAY ABOUT A LOT OF DATA WHICH HAS LED TO INFECTIOUS DISEASES IMAGING AND I SAW 6 DOTS WHERE THERE WAS HUMAN DATA INFECTION SPECIFIC. AND DEMA HAD ORGANIZED A MEETING AT THE NIH MORE THAN A DECADE AGO AND THEN WE ORGANIZED SEVERAL MEETINGS. WHAT I WANT TO POINT OUT IS THAT THERE IS A SUBSTANTIAL INCREASE IN INTEREST IN IMAGING OF INFECTIONS AND WE WANT TO MAINTAIN. I'LL SHOW MUTATA ON THAT AND THEN I'M GOING TO TALK ABOUT WHAT I THINK WOULD BE THE NEXT STEPS THAT WE NEED TO FOLLOW IN THIS FIELD. SO, AS EVERYBODY KNOWS, THERE IS IMFECKS OF IMAGING SUBGROUP -- INFECTION. DEMA LEADS THE IMAGING GROUP. AND WE HAVE BEEN ABLE TO DO A LOT OF THINGS UNDER THIS AND I REALLY WANT TO THANK THEM FOR THEIR SUPPORT. AND ONE OF THE THINGS I WANT TO POINT OUT IS THAT WE SHOULD ALL GET-TOGETHER. WE ARE A PRETTY SMALL GROUP. WE DON'T STILL HAVE CRITICAL MASS AND I THINK WE SHOULD SHARE AND HELP EACH OTHER IN DEVELOPING THIS FIELD. I SHOWED THIS SLIDE EARLIER BUT I WANTED TO HIGHLIGHT IT AGAIN AND I HAVEN'T DONE THIS WITH 2021 BUT THE NUMBER OF PUBLICATIONS AND INFECTIOUS DISEASE IMAGING SUBSTANTIALLY INCREASED AND I THINK WE SHOULD PAT OUR BACKS A BIT ABOUT THIS. I'M NOT TRYING TO SAY THAT EVERY TRACE OF DEVELOPING OR EVERY TRACE THAT IS GOING INTO HUMANS WILL BECOME A TRACER. BUT EVEN IF 1-10 BECOMES A TRACER WORKSY WILL HAVE SOME SOMETHING. AT THIS POINT WE HAVE ZERO. THERE IS NO PATHOGEN SPECIFIC IMAGING AT ALL AND I THINK THAT SOME OF THE WORK THAT WAS SHOWN TODAY ON BACTERIAL IMAGING AND THE VIRAL IMAGING WAS JUST AMAZING. IT'S BRILLIANT. NOT JUST ONLY IN HEALTHY DIAGNOSED AND MONITORED PATIENTS BUT IN PATHOGENESIS, HIV VIRAL RESERVOIRS, UNDERSTANDING THE BUG IS TAKEN CARE -- BACTERIA ARE TAKEN CARE OF BY THE ANTIBODIES. I WANT TO POINT OUT TWO QUICK THINGS AND I SAY THIS AT EVERY TALK. I THINK THAT THERE IS A PROBLEM IN OUR FIELD WHEN IT COMES TO RADIATION EXPOSURE. IN ON COLLIE, PEOPLE USE IMAGING RIGHT LEFT AND CENTER -- ONCOLOGY. SO IF YOU HAVE LYMPHOMA, WE ARE HAPPY TO DO A CT SCAN OR PET. BUT WE DON'T DO THE SAME THING WITH INFECTIONS. THIS IS DATA FROM A LOT OF SOURCES BUT WHAT I WANT TO SHOW YOU HERE IS THAT THE 5-YEAR MORTALITY RISK FOR A LOT OF DRUG RESISTANT INFECTIONS IS HIGHER THAN THE 5-YEAR MORTALITY RATE FOR A LOT OF CANCERS. WE HAVE TO BE PRAGMATIC. NOT EVERYBODY SHOULD BE SCANNED BUT WE SHOULD BE PRAGMATIC ABOUT IT AND THIS IS MORE RELEVANT TO INFECTIOUS DISEASES, PEOPLE WHO DON'T REALIZE WHAT THE RISKS ARE OF RADIATION. A RISK OF FLYING IN AN AIRPLANE AND WITH NEW TECHNOLOGIES AND TOTAL BODY PET ALSO SHOWN THIS RADIATION. THE OTHER THING I WANT TO POINT OUT IS THERE IS A LOT OF INTEREST IN INFECTIOUS DISEASES IMAGING AND DEVELOPING COUNTRIES COUNTRIES. AND WE REALLY NEED TO ENGAGE EVERYBODY. IT'S NOT JUST YOU IN AUSTRALIA, WE NEED TO GET ALL THE PEOPLE IN DEVELOPING COUNTRIES INTERESTED IN THIS. SOME OF THESE STUDIES WERE DONE IN THOSE COUNTRIES, HUMAN STUDIES. AND SO THAT IS SOMETHING ELSE. SO I THINK ONE OF THE THINGS THAT -- I THINK I FEEL VERY IMPORTANT IS AS WE TAKE THESE TRACERS INTO THE CLINIC, WE THEN NEED TO START ENGAGING THE STAKEHOLDERS AND I THINK THAT THAT IS GOING TO BE AN IMPORTANT THING. THIS HAS TO BE PICKED UP BY INDUSTRY OTHERWISE WE'LL PUBLISH NICE PAPERS BUT THAT IS WHERE IT IS GOING TO END. WE NEED INDUSTRY PEOPLE TO GET INTERESTED. THERE HAS TO BE PROFIT. WE HAVE TO SHOW HOW IMAGING CHANGES PATIENT MANAGEMENT AND SHOW HOW THIS IS A PRODUCT THAT IS WORTH FUNDING. SO I JUST WANT TO STOP HERE. I REALLY WANT TO THANK DEMA AGAIN FOR ORGANIZING THIS MEETING AND GETTING ME INTEREST. IF DEMA HAD NOT IN VETED ME TO THAT MEETING AT THE NIH, I WOULDN'T BE DOING ANYTHING. SO I'M GOING TO STOP AND FANTASTIC WORK BY EVERYBODY. WE ARE VERY PROUD AND WE WOULD LOVE TO SHARE ALL OUR KNOWLEDGE AND LOVE TO SHARE AND ENCOURAGE EVERYONE TO SHARE KNOWLEDGE WITH EACH OTHER. >> THANK YOU VERY MUCH, SANJAY. THAT WAS VERY NICE SUMMARY OF WHERE WE ARE AND WHERE WE WANT TO BE. I JUST WANT TO SAY THAT ON BEHALF OF ALL MY COLLEAGUES, COLLABORATORS AND ALL THE GUEST SPEAKERS, I WANT TO THANK YOU ALL FOR YOUR INTEREST AND PARTICIPATION TODAY. I KNOW IT WAS A VERY LONG DAY AND I REALLY HOPE YOU FOUND IT INTERESTING AND INFORMATIVE. AND MORE IMPORTANTLY, I REALLY HOPE IT INSPIRED SOME OF YOU TO JOIN US IN THIS GROWING FIELD BECAUSE THIS FIELD HAS SO MUCH TO EXPLORE AND DISCOVER. AND ALL TOGETHER WE CAN MOVE IT FORWARD. SO WITH THAT, I WOULD LIKE TO THANK YOU AGAIN AND HAVE A GREAT REST OF THE DAY.