GOOD AFTERNOON, EVERYBODY. FOR THOSE OF YOU WHO DON'T KNOW ME, I'M LARRY TABAK, AND I'M THE PRINCIPAL DEPUTY DIRECTOR OF NIH, AND FOR THOSE OF YOU WHO DO KNOW ME, USUALLY WHEN I SHOW UP TO AN EVENT, PEOPLE'S SHOULDERS SLUMP BECAUSE THEY REALIZE THAT FRANCIS ISN'T GOING TO BE HERE. BECAUSE WE'RE NEVER IN THE SAME PLACE AT THE SAME TIME. BUT FRANCIS UNFORTUNATELY IS DOWNTOWN, BUT EVEN IF HE WERE HERE, I STILL WOULD HAVE FOUGHT HIM TO MAKE THIS INTRODUCTION TODAY. SO I'M REALLY DELIGHTED TO BE ABLE TO DO THAT. IT'S NOT OFTEN THAT A KID WHO GRADUATED FROM CCNY CAN INTRODUCE A GUEST SPEAKER AT A WALS LECTURE WHO IS A DEAN AT THEIR ALMA MATER, BUT THAT IS EXACTLY WHAT I'M GOING TO HAVE THE PRIVILEGE OF DOING TODAY. SO DR. BARBRI KNOW DEAN OF THE GROVE SCHOO L OF ENGINEERING AT THE CITY COLLEGE OF NEW YORK, CCNY. SHE HAS APPOINTMENTS IN BIOMEDICAL ENGINEERING, CHEMICAL ENGINEERING AND THE CUNY SCHOOL OF MEDICINE. PRIOR TO JOINING CCNY, SHE WAS THE ASSOCIATE CHAIR FOR GRADUATE STUDIES AND PROFESSOR IN THE WALLACE H.KALTER DEPARTMENT OF ENGINEERING IN GEORGIA TECH AND EMORY, AND PRIOR TO GEORGIA TECH AND EMORY, SHE ROSE THROUGH THE RANKS AS FULL PROFESSOR OF CHEMICAL ENGINEERING AND SERVED AS VICE PROVOST FOR UNDERGRADUATE EDUCATION AT NORTHEASTERN. DR. BARABINO IS A NOTED EXPERT AND INVESTIGATOR IN THE AREAS OF SICKLE CELL DISEASE, CELLULAR AND TISSUE ENGINEERING, AND RACE ETHNICITY AND GENDER IN SCIENCE AND ENGINEERING. SO SHE HAS THIS REMARKABLE HYBRID APPROACH AND ACCOMPLISHMENT. SHE HAS WAY TOO MANY HONORS FOR ME TO ENUMERATE, BUT LET ME JUST GIVE YOU THE TOP LINE. SHE IS AN ELECTED MEMBER OF THE NATIONAL ACADEMY OF ENGINEERING, OF THE NATIONAL ACADEMIES, AND IS AN ELECTED FELLOW OF THE AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE, AND SHE HAVE IS THE FOUNDER AND EXECUTIVE DIRECTOR OF THE NATIONAL INSTITUTE FOR FACULTY EQUITY. IT IS AN ENORMOUS PLEASURE TO WELCOME YOU TODAY TO PROVIDE THIS AFTERNOON LECTURE. [APPLAUSE] >> GOOD AFTERNOON. I AM REALLY PLEASED AND HONORED TO BE HERE WITH YOU TODAY, AND I'D LIKE TO THANK ALL OF YOU FOR BEING HERE. I'D LIKE TO THANK THE ORGANIZERS AND THANK DR. TABAK FOR THAT KIND INTRODUCTION. TODAY I'D LIKE TO TALK TO YOU ABOUT SOME OF MY WORK, AND I'M GOING TO TELL YOU A LITTLE BIT ABOUT WHY I PICKED THIS TITLE, TWO ACT PLAY, THE CHARACTER OF CELLS AND THE ROLE OF BIOMECHANICS. I WANT TO TALK TO YOU ABOUT THAT IN THE CONTEXT OF THE RED BLOOD CELL, RED BLOOD CELL BIOMECHANICS AND IN THE CONTEXT OF SICKLE CELL DISEASE. THE RED BLOOD CELL IS A MODEL IN ITS STRUCTURE AND FUNCTION. IT'S A VISCOELASTIC MEMBRANE IN THE NORMAL STATE. IT'S BICONCAVE DISOIDAL SHAPE PERMITS THE MEMBRANE OF THE RED BLOOD CELL TO EXTEND AND HAVE THE MEMBRANE DEFORM WHILE IT'S MAINTAINING ITS CONSTANT SURFACE AREA AND WITHOUT STRETCHING. IT'S ALSO BEEN REFERRED TO AS A DUMBBELL SHAPE. SO THIS UNIQUE SHAPE OF THE SICKLE CELL -- EXCUSE ME -- OF THE NORMAL RED BLOOD CELL ALLOWS TO BEND OVER ON ITSELF AND PASS THROUGH VESSELS WITH A DIAMETER THAT'S ACTUALLY HALF OF THE DIAMETER OF THE RED CELL ITSELF. SO THAT'S THE FASCINATION AND THE UNIQUENESS OF A NORMAL RED BLOOD CELL. I'M GOING TO TELL YOU MORE ABOUT THE MECHANICAL PROPERTIES AND THE PROPERTIES OF A SICKLE CELL, BUT I'D LIKE TO GIVE YOU SOME SENSE FIRST OF SICKLE CELL DISEASE. IT AFFECTS MILLIONS WORLDWIDE, 100,000 AMERICANS, AND IT IMPOSES SIGNIFICANT PHYSICAL AND ECONOMIC BURDENS. NOW THERE'S TWO FDA-APPROVED DRUGS FOR SICKLE CELL DISEASE, AND THAT'S HYDROXYUREA AND MORE RECENTLY GLUTAMINE. SICKLE CELL DISEASE IS GLOBAL. HERE, YOU SEE A MAP THAT ACTUALLY SHOWS THE NUMBER OF NEWBORNS WITH SICKLE CELL ANEMIA IN 2015, AND WE KNOW THAT THE BIRTH RATE IS EXPECTED TO INCREASE BY 30% BETWEEN 2010 AND 2050. AND IF YOU NOTICE THE COLORATION, YOU CAN SEE IN THE DARKER AREAS IS THE INCREASED NUMBER OF NEWBORNS WITH SICKLE CELL DISEASE. I MENTIONED IT'S GLOBAL. I'D LIKE TO DRAW YOUR ATTENTION TO A MEETING THAT I ATTENDED THAT WAS AN INTERNATIONAL MEETING THAT BROUGHT TOGETHER WORLD EXPERTS ON SICKLE CELL DISEASE. IT WAS A PARTNERSHIP BETWEEN THE HOSPITAL FOR SICK CHILDREN IN TORONTO AND THE NATIONAL SICKLE CELL DISEASE CENTER IN THE REPUBLIC OF BENIN, AFRICA. THIS BUILDING YOU SEE HERE WAS THEIR SICKLE CELL TREATMENT CENTER. THIS IS A CLINIC FOR THOSE INDIVIDUALS LIVING WITH THE DISEASE AND OTHERS WERE COMING TO THIS AREA FOR TREATMENT. WE WERE ALSO THERE AT THIS MEETING ON THE OCCASION OF A GRAND OPENING FOR A NEW BUILDING, SO THIS NEW BUILDING IS JUST ACROSS FROM THE OLD BUILDING THAT YOU JUST SAW. AND THIS GRAND OPENING WAS FOR THE NATIONAL SICKLE CELL CENTER, IN JANUARY OF 2009. ONE OF THE THINGS THAT REALLY STOOD OUT IN THIS MEETING, BRINGING WORLD LEADERS IN SICKLE CELL DISEASE TOGETHER TO SPEAK SPECIFICALLY ABOUT RESEARCH AND EDUCATION AND PATIENT CARE FOR THE DISEASE WAS THAT THERE WERE AREAS THAT NEEDED TO BE ADDRESSED, AND THE TYPES OF THINGS THAT WERE NEEDED FOR BOTH DIAGNOSIS AND TREATMENT THAT WERE UNIFORM WORLDWIDE. AND ONE OF THE THINGS THAT STOOD OUT TO ME MOST AS I WAS THERE, AN INVESTIGATOR FROM THE UNITED STATES WORKING WITH THOSE FROM GLOBAL REGIONS, WAS -- PARTICULARLY IN AFRICA WAS THE LEVEL OF INCIDENCE OF THE DISEASE, THE LACK OF ACCESS TO DIAGNOSIS AND TREATMENT, AND HEARING SOME OF THE STORIES FROM INDIVIDUALS LIVING WITH THE DISEASE AND PARTICULARLY FROM MOTHERS WHO WERE IN THE VILLAGE AND OFTENTIMES WERE NOT HAVING ACCESS AND ABILITY TO BRING THEIR YOUNG CHILDREN IN, MANY TIMES THE CHILDREN WERE ACTUALLY PASSING AWAY BEFORE THEY HAD EVEN BEEN DIAGNOSED WITH THE DISEASE. THIS IS JUST ANOTHER FRONT OF THE BUILDING, THE NEW BUILDING WHERE THEY WERE STARTING TO HAVE THE NEW SICKLE CELL CENTER. THE LIFE EXPECTANCY IN SICKLE CELL DISEASE IS ROUGHLY ABOUT 30 YEARS LESS THAN THE NORMAL LIFE EXPECTANCY. WHAT I'D LIKE TO DRAW YOUR ATTENTION TO ON THIS PARTICULAR CHART, IF YOU LOOK AT THE INCREASES IN THE LIFE EXPECTANCY OVER TIME, YOU'LL SEE THAT THERE ARE A NUMBER OF ACTIVITIES OR INITIATIVES THAT TOOK PLACE THAT HAVE CONTRIBUTED TO THAT INCREASED LIFE EXPECTANCY. SO FOR EXAMPLE IN 1972, THERE WAS A NATIONAL SICKLE CELL ACT. PART OF THAT CONGRESSIONALLY MANDATED ACT WAS THE CREATION OF COMPREHENSIVE SICKLE CELL CENTERS TO HELP WITH THE TREATMENT AND THE EDUCATION AROUND THE DISEASE. THERE ARE ALSO THINGS THAT HAPPENED AROUND PREVENTIVE PENICILLIN, HYDROXYUREA BEING USED MORE UNIFORMLY FOR THE DISEASE, AND STRATEGIES AND THERAPIES LIKE HAVING TRANSFUSION FOR STROKE PREVENTION. THE DISEASE HAS A NUMBER OF CLINICAL MANIFESTATIONS, AND IF ONE IS LIVING WITH THE DISEASE, IT CAN BE EXTREMELY DEBILITATING. IT'S A MULTISYSTEM DISEASE WITH WIDESPREAD ORGAN DAMAGE. THE ORGAN DAMAGE CAN BECOME WORSE OVER TIME, AND YOU CAN SEE HERE THE DIFFERENT AREAS THAT ARE IMPACTED. FROM THE EYE TO THE LIVER AND THE GALLBLADDER AND SO ON. WE KNOW THAT THERE IS A GENETIC DEFECT, AND INTERESTINGLY, IT IS ONE AMINO ACID SUBSTITUTION IN THE BETA CHAIN, THE SIXTH POSITION IN THE CHAIN OF A HEMOGLOBIN MOLECULE THAT CAUSES THE HEMOGLOBIN TO PRELIMINARY RISE AND DEOXYGENATE IN CONDITIONS. BECAUSE I'M FOCUSING A BIT AND TRYING TO SHARE WITH YOU SOME OF THE INTERESTING PROPERTIES THAT ARE RELATED TO THE BIOMECHANICS, THE SICKLE CELL HEMOGLOBIN, PARTICULARLY WHEN IT PRELIMINARY RISES ACTUALLY DOES DESTROY THE MEMBRANE AND THE HEMOGLOBIN IN A PRELIMINARY RISED STATE CAN ACTUALLY PULL AT THE MEMBRANE, AND PULLING AT THE MEMBRANE, YOU CAN SEE HOW IT CAN BECOME DISTORTED AND MOVE TOWARDS THE SICKLE CELL SHAPE WHEN THE HEMOGLOBIN MOLECULES HAVE PRELIMINARY RISED. AND YOU CAN ALSO HAVE PROTEINS THAT ARE NORMALLY EMBEDDED IN THE MEMBRANE ABNORMALLY EXPOSED. I'D ALSO LIKE TO DRAW YOUR ATTENTION TO TWO PARTICULAR EXOAPTS OF SICKLE CELL DISEASE. ONE WE REFER TO AS A HEMOLYTIC COMPONENT, THE SICKLE CELLS ARE FRAGILE, THEY'RE EASILY DESTROYED, THEIR RAPID DESTRUCTION RESULTS IN HEE MOL CYST AND THUS THE HEMOLYTIC ANEMIA. THERE'S ALSO THE COMPONENT THAT'S VASOOCCLUSION WHICH CAN ALSO HAVE TWO COMPONENTS THAT'S RELATED TO A BLOCKAGE OF THE VESSELS RELATED TO PHYSICAL BLOCKAGE AND TRAPPING BECAUSE THE CELLS ARE STIFF AND CAN'T PASS THROUGH THE CIRCULATION, AS WELL AS AN ABNORMAL ADHESION OF THE SICKLE CELLS TO THE ENDOTHELIAL CELLS THAT LINE THE BLOOD VESSELS. SO DEPIBLGHTED DEPICTED HERE IS ONE COMPONENT WHERE YOU CAN HAVE CELLS THAT ARE MORE DEFORMABLE AND CAN ACTUALLY PASS THROUGH THE CIRCULATION A BIT AS SHOWN IN THE FIRST CASE, IF THERE'S STILL THE CAPACITY TO DEFORM. IF YOU HAVE A PARTIAL BLOCKAGE, IT COULD STILL PASS THROUGH THE CIRCULATION. BUT IN THE LATTER TWO CASES, THE OBSTRUCTION CAN ACTUALLY IMPEDE THE PASSAGE OF OTHER CELLS. AND ONE OF THE REASONS WHY WE WERE LOOKING AT ADHESION IS BECAUSE IF THE SICKLE CELLS ARE ABNORMALLY ADHERING TO THE BLOOD VESSELS, THEY CAN ACTUALLY IMPEDE THE PASSAGE OF OTHER CELLS THAT ARE NOT IN THE SICKLE SHAPE, AND CONTRIBUTE TO THE VASOOCCLUSIVE BLOCKAGE, AND THE VASOOCCLUSION, OF COURSE, IS A HALLMARK FOR THE DISEASE. SO IN LOOKING AT THE PATHOPHYSIOLOGY, WE JUST TALKED ABOUT THE HEMOGLOBIN MOLECULES CAN POLYMERIZE IN DEOXYGENATED CONDITIONS. IT'S ACTUALLY REVERSIBLE PROCESS. AND WHAT HAPPENS AFTER REPEATED SICKLING, YOU CAN HAVE PERMANENT DAMAGE, AND THE PERMANENT DAMAGE CAN LEAD TO WHAT WE REFER TO AS IRREVERSIBLY SICKLE CELLS, SO YOU CAN HAVE BLOCKAGE AGAIN RELATED TO THIS STIFFNESS OF THE CELLS, BUT WE ALSO HAVE THE COMPONENT VASO-OCCLUSION THAT'S CONTRIBUTED TO BY MULTICELL TYPES THAT CAN PARTICIPATE IN THE BLOCKAGE, ADHERING TO THE VESSEL WALLS, AS WELL AS CELLS ADHERING TO ONE ANOTHER, AND THE MECHANICAL TRAPPING OF CELLS AS WELL. SO YOU SEE HERE THAT YOU CAN HAVE PARTICIPATION BY BOTH THE WHITE CELLS AND THE RETICULOCYTES AND THE IRREVERSIBLY SICKLE CELLS INTERACTING WITH ENDOTHELIUM. SO NOW THAT YOU'VE GOTTEN A SENSE OF THE RED CELL AND ITS NORMAL PROPERTIES IN HEALTH, ITS UNIQUE SHAPE, THE BICONCAVE DISK, THE DEFORMABILITY AND ABILITY TO FOLD OVER ITSELF AND PASS OVER THE CIRCULATION, I'D LIKE TO PAY A BIT MORE ATTENTION TO THE ROLE IN DISEASE AND HERE YOU SEE THE HEMOGLOBIN MOLECULES THAT ARE AGGREGATING AND, HENCE, DISTORTING THE MORPHOLOGY AND SHAPE OF THE RED CELL, AND ITS ABILITY TO PASS THROUGH THE CIRCULATION. WE TIE WHAT WE KNOW ABOUT THESE MECHANICAL PROPERTIES SUCH AS THE STIFFNESS OF THE RED CELL, THE ABNORMAL ADHESIVENESS OF THE CELL, AND INCREASED VISCOSITY, WITH THE PATHOPHYSIOLOGY THAT WE KNOW, AND WE TRY TO SEE WHAT INSIGHTS WE CAN GET BY BETTER UNDERSTANDING THOSE BIOMECHANICS AND WHAT INSIGHTS IT MIGHT GIVE US TOWARDS THERAPIES, BOTH THERAPIES THAT EXIST AND THOSE THAT WE MAY WANT TO LOOK AT IN TERMS OF NEW STRATEGIES. THERAPIES THAT ARE TYPICALLY USED FOR SICKLE CELL DISEASE WOULD INCLUDE TRANSFUSION, I MENTIONED EARLIER HYDROXYUREA AND GLUTAMINE, WE'RE ALSO MOVING TOWARDS STRATEGIES THAT ARE CURATIVE SUCH AS GENE EDITING AND GENE THERAPY, AND OF COURSE THERE IS THE CURATIVE THERAPY OF BONE MARROW TRANSPLANTS. THERE'S SOME EARLY WORK THAT WAS DONE IN MY LAB WHERE WE WERE SPECIFICALLY LOOKING AT THE ABNORMAL ADHESION OF SICKLE RED BLOOD CELLS TO ENDOTHELIAL CELLS LINING THE BLOOD VESSELS, AND WE USED AN IN VITRO SYSTEM TO DO THAT ANALYSIS WITH A MICROSCOPY SYSTEM. SO THE FIGURE ON THE LEFT ACTUALLY SHOWS YOU WHAT HAPPENS IN TERMS OF ADHESIONS FOR INDIVIDUALS BEFORE THEY WERE ON HYDROXYUREA TREATMENT AND THEN AFTER. AND WE FOUND THAT WITHIN TWO WEEKS OF TREATMENT OF HYDROXYUREA, ADHESION WENT DOWN TO LEVELS OF NORMAL. WHAT YOU SEE ON THE RIGHT IN THE VIDEOS IS AN EXAMPLE OF A BLOOD SUSPENSION WITH SICKLE CELLS PASSING THROUGH ENDOTHELIALIZED MICRO CHANNELS WHERE THE LEFT IS AN INDIVIDUAL WHO IS ON HYDROXYUREA, OOND YOU'LL AND YOU'LL NOTIC E THE FLOW IS MUCH MORE SMOOTH AND FASTER, BUT IN THE CASE OF NOT BEING ON HYDROXYUREA, NOTE THE SLUGGISHNESS AND THE NATURE OF THE FLOW WHICH IS RELATED TO BOTH ABNORMAL ADHESION AND BLOCKAGE BECAUSE OF THE STIFFNESS OF THE CELLS. FURTHER LOOKING INTO EXPLOITING PROPERTIES THAT ARE RELATED TO MECHANICAL PROPERTIES OF THE CELL, WE LOOK AT THINGS LIKE THE RED CELL DENSITY AND SEPARATE THEM IN DIFFERENT POPULATIONS. THE POPULATIONS ARE LOOSELY RELATED TO THEIR DENSITY, AND THE DENSITY IS LOOSELY CORRELATED WITH AGE. SO THAT THE YOUNG CELLS THAT ARE LESS DENSE ARE THE RETICULOCYTES, AND THEN THE DISCOCYTES AND THE DENSE DISKOCYTES HAVE MORE DENSITY BUT TEND TO BE OLDER. THE ONES THAT ARE IRREVERSIBLY DAMAGED SICKLE CELLS TURN OUT TO BE THE MOST DENSE. THE REASON WHY WE'RE INTERESTED IN THESE PROPERTIES, WE'D LIKE TO KNOW IF WE CAN EXPLOIT THEM IN WAYS THAT WE COULD LEARN MORE ABOUT THE ABNORMAL FLOW IN SICKLE CELL DISEASE. IN THIS SIMULATION, THE RED CELLS ARE REPRESENTING CELLS THAT ARE NORMAL AND DEFORMABLE, AND LESS STIFF. THE BLUE CELLS ARE REPRESENTING THE STIFF CELLS THAT WILL REPRESENT SICKLE CELLS. AND WHAT WE KNOW ABOUT THE FLOW, THE NORMAL ONES THAT ARE MORE DEFORMABLE WILL TEND TO FLOW IN THE CENTER AND THE ONES THAT ARE LESS SO WILL MARGINATE OUT TO THE CHANNEL WALLS. THIS HAPPENS WHEN WE LOOK AT A MIXTURE THAT WE MIGHT BE ABLE TO EXPLOIT THAT DIFFERENCE IN THE STIFF CELLS ARE MARGIN NATEING AND THE MORE NORMAL MORE DEFORMABLE CELLS ARE NOT. SO THE WAY WE WOULD EMPLOY THAT IS TO SEPARATE CELLS BY THEIR STIFFNESS THAT WOULD ALSO BE CORRELATED WITH THINGS LIKE ARE THESE RI TICK LOW SITES OR ARE THESE MORE FERM NANTLY DAMAGED CELLS, AND THIS IS A DEVICE THAT'S SET UP SO IT COULD SEPARATE CELLS BY THE STIFFNESS. AND THE IDEA BEING IF YOU CAN HAVE THIS TYPE OF DEVICE, SEPARATE OUT BASED ON MECHANICAL PROPERTIES, CORRELATE THOSE MECHANICAL PROPERTIES WITH CELLS OF A PARTICULAR AGE OR DENSITY, WE MAY BE ABLE TO USE THAT AS A PROGNOSTIC DEVICE IF YOU TAKE A SAMPLE FROM AN INDIVIDUAL LIVING WITH THE DISEASE, CHARACTERIZE THEIR CELLS AND MAYBE USE THAT OVER TIME AS A WAY TO MONITOR THE PROGNOSIS OF THE DISEASE. SO I GAVE YOU SOME BACKGROUND AND INFORMATION ON JUST LOOKING AT THE RED CELL PROPERTIES AND HOW DO WE EXPLOIT THOSE PROPERTIES TO HAVE INSIGHTS INTO THE DISEASE. NOW I'D LIKE TO TURN YOUR ATTENTION TO LOOKING AT BONE OR TISSUE INVOLVEMENT THAT STILL IS RELATED TO THE DEFECT OF THE RED BLOOD CELLS. SO THE NEXT SET OF STUDIES THAT I'M GOING TO SHARE WITH YOU WERE ALL DONE WHERE WE WERE LOOKING TO CHARACTERIZE THE BONE PHENOTYPE IN SICKLE CELL DISEASE BASED ON MECHANICS. WE KNOW THAT THE PATHOLOGY IN SICKLE CELL BONE IS NOT WELL UNDERSTOOD. WE KNOW THAT THINGS LIKE JUST LOOKING AT BONE MINERAL DENSITY DOES NOT TELL THE WHOLE STORY. WE KNOW THAT IN SICKLE CELL DISEASE, THERE ARE SOME COMMONALITIES WITH OSTEOPOROSIS, THERE'S REDUCED MINERAL CONTENT AND REDUCED CORTICAL BONE THICKNESS, AND WE ALSO KNOW THAT THERE'S EXPANDED BONE MARROW CAVITY. AND PART OF THAT, WE BELIEVE, IS DUE TO THE FACT THAT YOU'RE IN A HEMO LYTIC STATE. THE HEMATOPOIETIC PROCESS IS IN OVERDRIVE. SICKLE CELL INDIVIDUALS ARE CONSTANTLY PRODUCING YOUNG RED BLOOD CELLS, THEIR PERCENTAGE OF RETICULOCYTES ARE MUCH HIGHER THAN NORMAL. WE ALSO KNOW FROM RADIOGRAPHS THAT THERE ARE THINGS LIKE IN SICKLE CELL A COMMON PHENOMENON THAT YOU SEE IN THE VERTEBRAE, THE COMPRESSION TAKES ON MORE OF AN "H" SHAPE. SO WE ARE, AS I MENTIONED, VERY MUCH INTERESTED IN GETTING A BETTER UNDERSTANDING OF WHAT MIGHT BE GOING ON. HOW TO YOU TIE IN THE PHYSIOLOGY OF THE HEMOLYSIS, THE -- THAT MIGHT BE PROMOTING REACTIVE OXYGEN SPECIES AND THE OCCLUSION THAT TAKES PLACE AS WELL. AND HOW THESE PROCESSES MIGHT RELATE TO THINGS LIKE AVASCULAR NECROSIS, NECROSIS OF THE BONE AND OTHER IMPAIRMENTS. SO IN THIS MODEL AND THE REST OF THE DATA THAT I'LL SHOW YOU, WE USE A TRANSGENIC MOUSE MODEL OF SICKLE CELL DISEASE, WHICH EXPRESSES HUMAN HEMOGLOBIN AND IT DEVELOPS A HEMOLYTIC ANEMIA AND THIS MODEL ACTUALLY DOES HAVE ORGAN DAMAGE THAT REPLICATES THE DAMAGE IN SICKLE CELL INDIVIDUALS. WE USE 10 WEEK OLD MICE TO REPRESENT ADOLESCENCE IN SICKLE CELL DISEASE, AND 21 WEEK OLD MICE TO REPRESENT THE ADULTS. WE USE THE FEMUR. THE FEMUR IN MOUSE MODELS HAS BEN USED TO APPROXIMATE HUMAN ACTIVITY. WE'VE CONCENTRATED ON LOOKING TETRABECK LAR AT THE TRABECHULAR BONE AND CORTICAL BONE. THIS IS CALLED A FOUR-POINT BENDING TEST, WHERE YOU PUT PRESSURE ON THE BONE UNTIL YOU GET BREAKAGE. AND AGAIN, THIS WAS WORK WE WERE LOOKING TO CHARACTERIZE THE BONE PHENOTYPE IN SICKLE CELL DISEASE. SO WHAT YOU SEE HERE IS, WE'RE LOOKING AT THE FEMORAL TRABECULAR BONE, THE BONE IS REDUCED IN TERMS OF ITS TRABECULAR THICKNESS AND THINNESS. SO 10 WEEKS HERE, 21 WEEKS, WE HAVE NORMAL, WE HAVE THE TRAIT AND WE HAVE SICKLE. THIS 3D MAPPING IS COLOR-CODED SO THAT RED ACTUALLY REPRESENTS THICKNESS. THICKEST. WHAT'S MOST NOTABLE, IF YOU LOOK AT SICKLE AND OLDER, YOU SEE THE MOST DAMAGED CELLS WHERE THIS BONE IS NOT NEARLY AS STRONG AS IT WOULD BE NORMALLY. SO THIS ONE, SAME TYPE OF MAP, BUT THIS ONE SHOWS YOU NORMAL TRAIT AND SICKLE FOR THE CORTICAL BONE. AND AGAIN, WE SEE THAT FOR SICKLE, YOU WOULD HAVE DIMINISHED THICKNESS AND IT CORRELATES WITH HAVING LESS STRENGTH. WE WERE ALSO INTERESTED IN KNOWING A BIT MORE ABOUT THESE MECHANICAL PROPERTIES THAT WE COULD LEARN BY BETTER UNDERSTANDING THE MICRO ARCHITECTURE. SO IN THIS CASE, WE WERE LOOKING AT MEASUREMENTS SPECIFICALLY LIKE STIFFNESS. THIS TOP IS 10 WEEKS AND THE LOWER ONE IS 21 WEEKS. SICKLE IS LESS STIFF. SICKLE IS LESS STRONG. BUT WHEN WE LOOKED AT A MEASUREMENT THAT SORT OF LOOKS AT PLIABILITY, IT'S CALLED POST YIELD DEFLECTION, SICKLE WAS A BIT HIGHER. AND THE IDEA IS THAT THERE MAY BE, EVEN THOUGH THERE'S MORE SPACES IN THE TRABECULAR BONE FOR SICKLE, FOR THE BONE THAT'S THERE, IN THOSE -- IN BETWEEN THE VOID SPACES, IT MAY ACTUALLY BE A BIT MORE PLIABLE. THAT'S ONE OF THE THINGS THAT WE THINK MAY BE HAPPENING BECAUSE IN INDIVIDUALS LIVING WITH THE DISEASE, THERE ARE A NUMBER OF COMPENSATORY MECHANISMS THAT TAKE PLACE. WE ALSO LOOKED AT SOME HISTOLOGY SPECIFICALLY IN THE BONE MARROW, LOOKING FOR ADDITIONAL CLUES AS TO WHAT MIGHT BE HAPPENING. AND LOOKING FOR OSTEONE CRATIC FEATURES. IN THIS CASE, THIS AMORPHOUS REGION FOR -- IN THE SICKLE MARROW WAS SEEN CONSISTENTLY FOR THE SICKLE MICE, BUT IT WAS NOT SEEN IN THE NORMAL CASES. SO WE'RE WORKING TO HAVE A BETTER UNDERSTANDING OF WHAT MIGHT ACCOUNT FOR THOSE DIFFERENCES IN THE MARROW FOR SICKLE VERSUS NORMAL. SO JUST TO SUM UP BEFORE MOVING ON TO THE NEXT SECTION, THE MAIN POINT WAS THAT THE SICKLE BONE QUALITY IS DECREASED VERSUS NORMAL, AND IT'S EXACERBATED WITH AGE SO THE SICKLE BONES ARE SIGNIFICANTLY DETERIORATED AND THERE'S LESS TRABECULAE AND WHAT'S THERE IS THINNER. I MENTIONED THAT THERE ARE TWO DRUGS THAT HAVE BEEN USED, GLUTAMINE MORE RECENTLY, SO GLUTAMINE WAS APPROVED FOR SICKLE CELL USE IN 2017. WE WANTED TO EXAMINE IN THE LABORATORY HOW EFFECTIVE MIGHT GLUTAMINE BE AS A THERAPY BY USING THE MODEL THAT WE JUST STUDIED FOR BONE TO SHOW THE BONE PHENOTYPE IN SICKLE CELL DISEASE, TO SEE IF THAT MIGHT ACTUALLY BE AN APPROPRIATE MODEL TO EXAMINE A THERAPY. AND IN THIS CASE, WE'RE SPECIFICALLY LOOKING AT THE INFLUENCE OF GLUTAMINE ON SICKLE BONE MECHANICS. WE HAD SICKLE MICE, THEY WERE MALE MICE, WE HAD CONTROLS AND THOSE SAME TOWNS MICE WHO HAD SICKLE CELL DISEASE, THE GLUTAMINE WAS ADDED TO THE DRINKING WATER, AND WE DID SIMILAR TESTS WITH THE TRABECULAR BONE AND THE CORTICAL BONE TO LOOK AT MECHANICAL PROPERTIES. INTERESTINGLY, WE SAW THAT GLUTAMINE WAS HAVING AB AN EFFECT, AND ONE OF THE THINGS WE THOUGHT WE'D LOOK AT IN PARTICULAR WAS BECAUSE OF THE END ORGAN DAMAGE THAT YOU SEE IN SICKLE CELL DISEASE, AND BECAUSE THE SPLEEN IS ACTUALLY ONE OF THE FIRST ORGANS TO INDICATE THE DAMAGE RELATED TO SICKLING, WE WANTED TO LOOK AT SPLEEN SIZE. SO WHAT YOU SEE HERE ARE THE SPLEENS FOR THE SICKLE MICE, WHEN THERE'S TREATMENT WITH GLUTAMINE, THE SPLEEN SIZE ACTUALLY IS DECREASED. AND WE WANTED TO HAVE A BETTER SENSE OF WHAT IS THE IMPACT OF GLUTAMINE. WE ALSO NOTICE, AND WHAT WE SHOW AS GLUTAMINE PLUS OR GLUTAMINE MINUS, IN SOME OF THE MICE HAVING THE GLUTAMINE ACTUALLY CAUSED THE SPLEEN TO DECREASE IN SIZE AND IMPROVE BONE PROPERTIES, AND WE REFER TO THOSE AS GLUTAMINE PLUS, BUT IN SOME MICE, THERE WASN'T NECESSARILY A POSITIVE EFFECT, EVEN THOUGH THEY HAD GLUTAMINE TREATMENT VERSUS NOT TREATMENT, AND WE REFER TO THOSE AS GLUTAMINE MINUS. SO JUST LOOKING AT WHAT HAPPENED TO THE TRABECULAR BONE, AT THE DIFFERENT AGES, THIS DIAGRAM IS ILLUSTRATIVE OF THE FACT THAT WE'VE SEEN, THIS IS WILD TYPE CONTROL, THIS IS SICKLE. THE GLUTAMINE WITH THE POSITIVE EFFECT, YOU CAN SEE THAT THERE'S AN INCREASE IN THE BONE VOLUME. NOTING THAT, WE WERE INTERESTED TO BETTER UNDERSTAND WHAT MIGHT BE CONTRIBUTING TO THIS IMPROVEMENT IN BONE FOR SICKLE, SO WE LOOKED AT OSTEOGENIC MARKERS, SO A MARKER FOR OSTEOGENIC BEHAVIOR WHERE THE OSTEOBLASTS -- OSTEOBLAST WITH BONE FORMATION AND THEN WE LOOKED AT OSTEOCLAST FOR RESORPTION. SO USING THESE MARKERS, IT'S CLEAR THAT IN RESPONSE TO GLUTAMINE, YOU CAN HAVE INCREASES IN OSTEOGENIC ACTIVITY, BUT FOSH FOR THE OSTEOCLAST RESORPTION ACTIVITY, WE'RE ACTUALLY SEEING DECREASES WITH RELATION TO THE GLUTAMINE TREATMENT. WE WERE ALSO INTERESTED IN BETTER UNDERSTANDING WHAT HAPPENS AND WHAT MIGHT BE CONTRIBUTING TO WHAT WE'VE SEEN WITH THE SPLEEN AND THE BONE. SO INSULIN GROWTH FACTOR 1, IGF-1, IS KNOWN TO ENHANCE BONE TISSUE DEVELOPMENT AND GROWTH AND IT'S ALSO KNOWN TO BE PRODUCED IN THE LIVER. SO WE'VE WANTED TO GET A BETTER SENSE OF WHAT HAPPENS IN RESPONSE TO THE IGF LEVELS, AND YOU CAN SEE THAT IGF DOES MAKE A DIFFERENCE IN THE BONE WITH THE -- WE WERE LOOKING FOR THE ONES THAT -- THE MICE THAT ACTUALLY HAD SMALL SPLEENS, AND THERE SEEMS TO BE INCREASED IGF-1 EXPRESSION. SO WE ALSO LOOKED AT ANOTHER LIVER PROTEIN TO SEE IF IT WAS INCREASED IN RESPONSE TO GLUTAMINE, AND WE LOOKED AT HEMOPEXIN PARTICULARLY BECAUSE IT'S PRODUCED IN THE LIVER AND IT SERS AS SERVES AS A SCAVENGER WHEN THERE'S FREE HEME, AND AGAIN, WE CAN SEE THAT THE GLUTAMINE THERAPY HAD AN EFFECT. AND AGAIN, WE'RE LOOKING AT THE MICE FROM 4 TO 16 WEEKS OF AGE, AND WE ALSO NOTICE THAT WHEN THE MICE WERE AT 16 WEEKS OF AGE, WHICH WE ARE CALLING SKELETAL MATURITY, YOU MAY NOT SEE AS DRAMATIC OF AN EFFECT. AND FINALLY, WE WERE LOOKING AT WHAT IS THE IMPACT OF HAVING GLUTAMINE THERAPY ON THE BONE MARROW AND WE LOOKED FOR IRON DEPOSITS. SO THIS PRUSSIAN BLUE STAINING WAS FOR IRON DEPOSITS, AND IT SEEMS THAT GLUTAMINE THERAPY CAN ACTUALLY INCREASE THE IRON DEPOSITS AND ONE OF THE REASONS WHY WE WERE INTERESTED IN THIS IS BECAUSE OF IRON DEPOSITS, AND IN SICKLE CELL DISEASE AS IT RELATES TO ANEMIA. THE HISTOLOGY THAT WE WERE USING FOR BONE MARROW ACTUALLY GAVE US MORE INDICATIONS ABOUT WHAT MIGHT BE HAPPENING FOR SICKLE VERSUS NORMAL. AND INTERESTINGLY, WE SAW THAT CONSISTENTLY FOR THE SICKLE MICE WITH GLUTAMINE TREATMENT, WE'D SEE THESE OPEN SPACES THAT ARE VERY HARD TO DESCRIBE WHY THEY ARE THERE, BUT THEY WERE CONSISTENT. WE SAW THESE IN ALL OF THE MICE, WE DON'T HAVE AN IMMEDIATE EXPLANATION FOR THOSE BUT THAT'S ONE OF THE THINGS THAT WE'LL BE LOOKING INTO. BUT THE CLEAR IMPLICATIONS HERE, IS THAT HAVING SICKLE CELL DISEASE IMPACTS BONE, BONE PROPERTIES, AND THERE'S ALSO SOME INFLUENCE AS TO WHAT'S GOING ON IN THE BONE MARROW. SO I WANTED TO SUM UP A LITTLE BIT ABOUT SOME OF THE FINDINGS THAT WE HAD THAT WERE SPECIFICALLY RELATED TO THE INSIGHTS THAT WE WERE ABLE TO GET BY USING GLUTAMINE TREATMENT AND LOOKING AT BONE. WE SAW SOME PROTECTIVE EFFECTS OF THE GLUTAMINE, THOUGH THERE WAS A POPULATION WHERE THE GLUTAMINE DID NOT HAVE AN EFFECT AND IT SEEMED TO HAVE SOME RELATIONSHIP WITH THE SKELETAL MATURITY OF THE BONE. WE ALSO SAW THAT GLUTAMINE COULD INCREASE THE LIVER PROTEIN EXPRESSION OF CERTAIN PROTEINS AND HORMONES LIKE IGF-1, AND THAT GLUTAMINE DOES APPEAR TO TO INFLUENCE BONE CELL PHYSIOLOGY IN FAVOR OF BONE FORMATION AS OPPOSED TO BONE RESORPTION. IT'S CLEAR THAT WE'RE GOING TO NEED TO DO ADDITIONAL WORK TO GET A BETTER UNDERSTANDING OF WHAT ARE THE MECHANISMS THAT CONTRIBUTE TO THIS AB ABNORMAL PHENOTYPE OF BONE IN SICKLE CELL DISEASE, AND WE'RE HOPING THAT FURTHER STUDIES WITH A TREATMENT LIKE GLUTAMINE COULD HELP US GAIN ADDITIONAL INSIGHTS AS TO WHAT MIGHT BE HAPPENING IN THIS MOUSE MODEL THAT'S TRANSLATABLE THAT WOULD BE USEFUL FOR TREATING HUMANS. BEFORE I END, I'D LIKE TO TELL YOU BIT ABOUT SOME FUTURE ACTIVITIES AND DIRECTIONS THAT ARE HAPPENING NOT JUST IN THE RESEARCH ARENA ARENA BUT AROUND SICKLE CELL DISEASE. NIH HAS AN INITIATIVE ON SICKLE CELL DISEASE, "CURE SICKLE CELL." IT'S A VERY AMBITIOUS NIRTIVE, BUT INITIATIVE, BUT IMPORTANTLY IT MAKES CONNECTIONS BETWEEN PROFESSIONAL SOCIETIES AND COMPANIES AND ACADEMIC INSTITUTIONS, PATIENTS AND ADVOCACY GROUPS AND PROVIDERS AND RESEARCHERS AND OTHER AGENCIES. IT IS ALSO PARTNERED IN PARTICULAR WITH THE AMERICAN SOCIETY OF HEMATOLOGY. WHAT'S IMPORTANT ABOUT THIS TYPE OF COMPREHENSIVE INITIATIVE IS THAT WE'RE DEALING WITH A VERY COMPLEX DISORDER. THERE ARE IMPLICATIONS THAT ARE PUBLIC HEALTH IMPLICATIONS, THERE ARE IMPLICATIONS AROUND PSYCHOSOCIAL, SOCIOECONOMIC. YOU NEED A VERY LARGE MULTIPRONGED, MULTIFACETED APPROACH. ASH, AMERICAN SOCIETY OF HEMATOLOGY, IN ADDITION TO COLLABORATEING WITH NIH AND HOARS OTHERS HAS ITS OWN INITIATIVE LIKE CREATING ACCESS TO CARE, CREATING COLICKSES, COALITIONS, TRAINING F OR INDIVIDUALS LIVING WITH THE DISEASE, THEIR FAMILIES AND SUPPORTERS AND PROVIDERS AS WELL, LOOKING INTO ISSUES LIKE RESEARCH IN CLINICAL TRIALS AND WHO HAS ACCESS TO THEM, AND LOOKING AT THE GLOBAL FOOTPRNT, AND THE GLOBAL IMPACT OF HOW DO WE TREAT AND MANAGE THE DISEASE. THE NATIONAL ACADEMIES ACTUALLY HAS HAD ITS OWN STUDY. I'M A MEMBER OF THE STUDY COMMITTEE WHERE OUR CHARGE WAS ADDRESSING SICKLE CELL DISEASE, A STRATEGIC PLAN AND BLUEPRINT FOR ACTION. WE FINISHED OUR WORK IN SEPTEMBER AND A REPORT ON THAT WORK IS FORTHCOMING. ONE OF THE THINGS THAT WAS VERY IMPORTANT ABOUT THIS WORK AS WELL, IT WAS ALSO A VERY COMPREHENSIVE APPROACH TO BRING TOGETHER THE INFORMATION THAT COULD BE GAINED AND GLEANED FROM INDIVIDUALS LIVING WITH THE DISEASE, ADVOCATES, COMMUNITY GROUPS, AGENCIES, PROVIDERS, THE HEALTHCARE INDUSTRY, THOSE WHO ARE FUNDING OR NOT FUNDING WHEN YOU ARE BEING REIMBURSED FOR HEALTHCARE AND SO ON. SO THIS IS A VERY IMPORTANT ACTIVITY, AND I URGE YOU ALL TO LOOK AT THIS REPORT WHEN IT COMES OUT. I'M SURE YOU'VE SEEN OTHER WAYS THAT SICKLE CELL DISEASE HAS GOTTEN ATTENTION. THIS SET OF PICTURES IS FROM A 60 MINUTES SPECIAL THAT WAS DONE, AND THIS 60 MINUTES SPECIAL WAS SPECIFICALLY TALKING ABOUT THE CLINICAL TRIALS THAT WERE HAPPENING HERE AT NIH. MOST RECENTLY, THIS IS TAKEN FROM AN ARTICLE FROM "THE NEW YORK TIMES," JANUARY 11TH. THIS IS TALKING ABOUT THE YOUNGEST INDIVIDUAL LIVING WITH SICKLE CELL DISEASE TO HAVE THE GENE THERAPY. AND IN THIS PARTICULAR ARTICLE, THEY CHRONICLE HER STORY. SO THIS CURATIVE APPROACH HAS GREAT PROMISE, BUT IT ALSO HAS LOTS OF UNKNOWNS, AND THERE'S LOTS OF TREPIDATION AROUND HOW DO WE MOVE FORWARD. BUT WE'RE CERTAINLY MOVING FORTH GREAT SPEED AND ATTENTION TO A DISEASE AS DEBILITATING AS SICKLE CELL DISEASE, AND I THINK IT'S VERY IMPORTANT THAT AN AGENCY LIKE NIH IS PLAYING A MAJOR ROLE IN LEADING THAT EFFORT. SO I'D LIKE TO CLOSE BY DRAWING YOUR ATTENTION BACK TO WHERE WE STARTED. THAT IF WE BETTER UNDERSTAND CELL AND TISSUE BIOMECHANICS, THEY ACTUALLY DO PROVIDE TELLTALE SIGNS OF DISEASE, IT CAN SERVE AS A DISKRIM NAY TORE DISCRIMINA TE R BETWEEN WHAT'S HEALTH AND DISEASE AND WE CAN ALSO GET A ACCEPTS OF THE INDICATORS OF THE EFFECTIVENESS OF THE THERAPIES, AS I SHOWED YOU WITH THE WORK WE DID WITH THE SICKLE MOUSE MODEL AND LOOKING AT THE INFLUENCE OF GLUTAMINE. AND LASTLY AS I THANK YOU FOR YOUR ATTENTION, I'D LIKE TO DRAW YOUR ATTENTION BACK TO SICKLE CELL DISEASE, THE IMPORTANCE OF UNDERSTANDING IT, AND INDIVIDUALS LIVING WITH THE DISEASE AND THE COMPLICATIONS. SO THIS DRAWING IS BY A HAITIAN-BORN ARTIST WHO'S LIVING WITH SICKLE CELL DISEASE AND HAS USED ART AS A WAY OF EXPRESSING AND FOR HIM, COPING WITH LIVING WITH THE DISEASE. IN THIS PARTICULAR PICTURE, HE WAS DEPICTING A PAINFUL VASO-OCCLUSIVE CRISIS. I'D LIKE TO JUST END WITH THE IMPORTANCE OF THE WORK THAT WE ALL DO WHERE WE BRING DIFFERENT PERSPECTIVES TO UNDERSTANDING A MEDICAL PROBLEM, AND LOOK AT THINGS LIKE SOMETHING AS NUANCED AS A RED CELL AND ITS BIOMECHANICS PROPERTIES, ITS CHARACTER, AND HOW THAT MIGHT TELL US SOMETHING ABOUT A DISEASE AND HOW TO BETTER UNDERSTAND IT, AND PUT TOGETHER EFFECTIVE STRATEGIES TOWARDS TREATMENT AND TOWARDS CURES. SO THANK YOU VERY MUCH FOR YOUR ATTENTION. [APPLAUSE] >> THANK YOU VERY MUCH. WE HAVE TIME FOR SOME QUESTIONS. I WOULD ASK THAT PEOPLE COME TO ONE OF THE TWO MICROSCOPE -- MICROPHONES SO THAT PEOPLE WHO ARE WATCHING BY TELECONFERENCE ARE ABE TO HEAR WHAT THE QUESTIONS ARE, AND I'LL GET THINGS STARTED BY ASKING THE QUESTION. YOU DID SOME REALLY ELEGANT EXPERIMENTS TO LOOK AT THE DEFORMITIES OF THE RED CELLS IN THIS PARTICULAR DISEASE CONDITION. CAN YOU TELL US HOW THAT MIGHT END UP BEING DIAGNOSTIC FOR THOSE PEOPLE WHO ARE GETTING GENE THERAPY? DID IT SEEM -- HOW WELL THAT'S IMPACTED THEM OR WHEN THEY HAVE A RELAPSE, AND WHAT OTHER SORTS OF DISEASE CONDITIONS MIGHT THIS ALSO BE APPLIED TO? >> SURE. SO THE KINDS OF ASSAYS OR THE LOOKING AT MECHANICAL PROPERTIES THAT WE HAVE, THE IDEA IS THAT, SO YOU CAN LOOK AT WHAT THE CELLS LOOK LIKE NORMALLY. THEN IN THANTHEN IN A DISEASE STATE, THEN IN RESPONSE TO THERAPY. SO WE'VE CHARACTERIZED THIS AND DONE THIS IN THESE DIFFERENT LEVELS. ONCE YOU GET TO A STATE WHERE THE INDIVIDUAL IS CURED, WHAT DOES THAT CURE LOOK LIKE? WE CAN DO THESE SAME KINDS OF ANALYSES AND ASSAYS WITH THEIR BLOOD OVER A PERIOD OF TIME TO SEE HOW THAT COMPARES TO A NORMAL STATE, FOR EXAMPLE. SO THAT'S THE IDEA THAT WE WOULD STILL DO THESE SAME KINDS OF CHARACTERIZATIONS, AND USE MECHANICS IN THAT WAY. SO THAT'S WHERE WE'RE HEADED WITH IT. IN TERMS OF OTHER THINGS THAT YOU CAN USE IT FOR, THERE ARE OTHER CONDITIONS WHERE A CELL, PARTICULARLY A RED CELL, MIGHT BE -- MIGHT HAVE PROBLEMS OR MIGHT BE COMPROMISED MECHANICALLY, AND SO YOU CAN STILL USE WHAT YOU'VE LEARNED IN A SITUATION LIKE SICKLE CELL DISEASE, FOR EXAMPLE, IN ANOTHER DISEASE STATE, WHERE YOU MIGHT HAVE SOME STIFFNESS OF A CELL. SO THERE'S SOME THINGS THAT WE'VE BEEN DOING IN COMPARING LIKE SICKLE CELL -- A SICKLED RED BLOOD CELL VERSUS A CELL THAT'S A MALARIA CELL. ANYTHING WHERE THE DISEASE OR THE CONDITION IMPACTS THE STIFFNESS, FOR EXAMPLE, THAT'S ANOTHER WAY YOU CAN APPLY THESE TECHNIQUES AND TECHNOLOGIES FOR ANOTHER DISEASE STATE. >> HELLO! >> THANK YOU. AND HAVE YOU RUN THESE STUDIES COMPARING THE DIFFERENT GENOTYPES LIKE SICKLE BETA 0THAL AND OTHER ANEMIAS VERSUS SC DISEASE? HAVE YOU SEEN A DIFFERENCE IN THE DEFORMABILITY? >> WE HAVE NOT IN OUR MOUSE STUDIES. BUT IN SOME OF OUR EARLIER WORK WHERE WE WERE JUST LOOKING -- USING FLOW CHANNELS AND LOOKING AT ADHESION FOR SICKLE, WE HAVE HAD CASES WHERE WE WERE LOOKING AT HOMOZYGOUS OR NON-HOMOZYGOUS AND MAYBE LIKE SOMEONE THAT HAD THALASSEMIA ALONG WITH IT. USUALLY, THE PRESENCE OF SOMETHING ELSE BEYOND JUST HOMOZYGOUS SICKLE, YOU TEND TO SEE LESS ADHESION, AND SO WE ALSO DID THINGS LIKE WE LOOKED AT SICKLE TRAIT, SO SICKLE TRAIT WOULD TEND TO BE SOMEWHERE IN THE MIDDLE, LIKE YOU WOULD THINK IT WOULD BE TOTALLY NORMAL BUT NOT NECESSARILY NORMAL BECAUSE THERE ARE SOME ASPECTS OF SICKLE TRAIT, AND ONE OF THE DISEASE STATES I LOOKED AT EARLY ORN WAS HEREDITARY SPHERE OWE CYTOSIS. THE REASON WHY I WAS LOOKING AT THAT ONE IS THEY TEND TO HAVE A HIGH PERCENTAGE OF RETICULOCYTES AS SICKLE CELL PATIENTS DO, AND THEY ALSO HAD HIGHER ADHESION THAN NORMAL, BUT THEN YOU WOULD ASK, WHY DON'T THEY HAVE THE SAME KINDS OF COMPLICATIONS THAT YOU WOULD SEE IN SICKLE? AND IT'S BECAUSE THEY DON'T HAVE -- FOR HEREDITARY SPHEROCYTOSIS, THEY DON'T HAVE HEMOGLOBIN THAT HAS ALL THOSE COMPLICATIONS. SO IT WASN'T POSING THE SAME KINDS OF PROBLEMS. BUT I THINK IT IS VERY IMPORTANT TO LOOK AT THE OTHER PHENOTYPES AND NOT JUST THE HOMOZYGOUS. >> THERE'S A SIMILAR PROBLEM WITH RESPECT TO METASTASES, WHEN YOU GET WHAT'S CALLED THE EMT, THE EPITHELIAL MESENCHYMAL TRANSITION. IT'S THOUGHT WHEN YOU HAVE A CANCER AND IT'S AN EP PEEL YAL CELL SO THE QUESTION IS, HOW DOES IT GET OUT OF THE ORGAN AND GO TO THE BLOODSTREAM. ONE OF THE HYPOTHESIS THERAPIES ARE BASED ON NOW IS IT TURNS INTO A ME SYNC MALL CELL WHICH IS KIND OF THE SAME DEAL YOU'RE TALKING ABOUT HERE AND THEN KIND OF SQUEEZES THROUGH INTO THE CIRCULATION BECAUSE NOW IT'S GOT A MALLEABLE SHAPE, BUT WHAT I'M DRAWING THERE, YOUR ENGINEERING CONCEPT ALSO APPLIES TO THE FIELD OF METASTASES, WHICH IS MORE COMPLICATED THAN MOST PEOPLE IMAGINE, THE EMT TRANSITION. DO YOU HAVE ANY EXPERIENCE WITH THAT? >> I DON'T. BUT WHAT YOU'RE SAYING, WHAT WE'RE SEEING IS TOTALLY ANALOGOUS TO THE BEHAVIOR THAT YOU JUST DESCRIBED. BECAUSE WHEN A CELL IS MORE DEFORMABLE, IT CAN SQUEEZE, AND WHEN IT'S MORE STIFF, IT'S GOING TO DO ALL SORTS OF THINGS THAT IT CAN'T DO JUST BECAUSE OF THE STIFFNESS, SO THAT'S SOMETHING THAT WE SHOULD LOOK AT AS WELL. >> AND ALSO A GENERAL CONCEPT THAT I'VE SEEN, I'M A PA PATHOLOGIST AND MANY YEARS AGO I REMEMBER LOOKING AT A HEART AND SEEING -- AT THE HISTOLOGY AND SEEING ACTUALLY BONE FORMATION, AND THIS BRINGS UP THE CONCEPT OF THE HUMAN TISSUES IN TERMS OF META MALACIA, IS WHAT WE PLASIA, MEANING O NE CELL CAN TRANSITION INTO EACH OTHER. YOU CAN GET IT ANYWHERE, CELLS HAVE THAT CAPACITY. SO THIS ALSO, I'M SURE, HAS IMPLICATIONS FOR THE TYPE OF ENGINEERING -- IN OTHER WORDS, IF YOU SMACK A CELL AROUND ENOUGH JUST RANDOMLY, IT CONVERTS TO A DIFFERENT TISSUE. SO I DON'T KNOW HOW THAT WOULD INCORPORATE DIRECTLY INTO THIS, BUT I JUST BROUGHT THAT UP AS A POINT OF INTEREST. >> BUT THAT'S FASCINATING, AND I WOULD LOVE TO FOLLOW UP ON SOME OF THAT BECAUSE PART OF WHAT WE DO IN MY LAB IS LOOK AT THE IMPACT/INFLUENCE OF ANY TYPE OF FORCE. SO THE IDEA THAT YOU CAN HAVE A PHYSICAL FORCE, A FLUID FORCE, THAT WOULD IMPACT THE CELLS' BEHAVIOR IS REALLY IMPORTANT. SO THAT CONCEPT IS VERY MUCH WORTH PURSUING. >> YOU SHOULD REALLY LOOK INTO THE EMT. IT'S BIG IN THE FIELD OF METASTASIS. THANK YOU VERY MUCH. >> I WILL LOOK INTO THAT. THANK YOU. >> HI, MY NAME IS JANET JOSEPH. SO I SEE THAT A LOT OF THE INTERVENTIONS THAT YOU HAVE THAT IS OUT THERE RIGHT NOW ARE FOR ADOLESCENTS. SO I WAS JUST THINKING, YOU TALKED ABOUT BONE AND DEGENERATION. HAVE YOU BEEN LOOKING INTO HALTING THE DEGENERATION PROCESS IN SOME WAY OR REVERSING OR MAYBE THINKING OF WAYS TO REVERSE THE DEGENERATION PROCESS? BECAUSE I'M THINKING THAT INDIVIDUALS WHO ARE A LITTLE OLDER, YOU KNOW, LIKE HOW CAN THEY SURVIVE, OR WHAT ARE THE INTERVENTIONS THAT ARE OUT THERE FOR THEM. >> SO WE HAVE NOT TO DATE, BUT THAT'S EXACTLY WHERE WE'RE HEADED. SO THE FIRST THING WE THOUGHT WAS THERE WAS SO LITTLE CHARACTERIZED ABOUT THE BONE, ESPECIALLY FOR MECHANICAL PROPERTIES, THAT WE FIRST HAD TO DEMONSTRATE A BONE PHENOTYPE THAT THERE REALLY WAS A DIFFERENCE IN SICKLE VERSUS NORMAL. AND THEN WE SAID, OKAY, WE DEMONSTRATED THIS, HOW DO WE ACCOUNT FOR IT, LIKE WHAT'S THE MECHANISM BEHIND IT. SO NOW WE ACTUALLY NEED TO DO MORE MECHANISTIC STUDIES, AND HAVE A BETTER SENSE OF THE BONE REMODELING PROCESS, WHAT CELLS ARE PARTICIPATING WHEN, BECAUSE IN THE BODY, THERE'S HOMEOSTASIS AROUND REMODELING, THE BONE IS CONSTANTLY REMODELING. WITH ALL OF THE DEFECTS IN SICKLE CELL DISEASE, AND THIS IS WHY I WAS THINKING WE WOULD HAVE -- THERE'S NO WAY WE WOULD BE ABLE TO DEMONSTRATE SOME KIND OF PHENOTYPE FOR SICKLE BONE, BECAUSE FOR ALL OF WHAT'S GOING ON IN SICKLE CELL DISEASE, YOU CAN'T TELL ME THAT THE BONE REMODELING PROCESS IS NOT AFFECTED AS WELL. AND IF WE CAN BETTER UNDERSTAND THAT, I THINK WE COULD GET TO SOME OF THE THINGS THAT YOU'RE TALKING ABOUT, BECAUSE ONCE YOU UNDERSTAND THAT PROCESS, HOW DO YOU THEN INTERRUPT IT. ONE OF THE REASONS WHY WE WERE VERY INTERESTED IN STUDYING BONE MORE EXPLICITLY, INDIVIDUALS LIVING WITH THE DISEASE, EVEN THE YOUNG ONES, TALK A LOT ABOUT BONE PAIN, LIKE THIS DEEP BONE -- IT'S IN YOUR BONES. AND WE'RE LIKE, HOW CAN WE NOT BETTER UNDERSTAND WHAT'S GOING ON WITH THE BONE. AND I ALSO THINK A WIDE OPEN AREA IS WHAT IS GOING ON IN THAT BONE MARROW. PARTICULARLY FOR INDIVIDUALS LIVING WITH THE DISEASE. AND THE IDEA THAT IF THE HEMATOPOIETIC PROCESS IS IN OVERDRIVE, THE CAVITY IS ONLY SO BIG, AND THE KIND OF PRESSURE THAT MIGHT BE THERE, OR NERVE ENDINGS. WHEN YOU GET INFARCTS IN THE BONE AND THE BONE MARROW, SO I THINK THAT'S WHERE WE'RE HEADED, TO HAVE A BRETTER BETTER UNDERSTANDING ME MECHANISTICALLY SO WE CAN FIGURE OUT HOW TO BETTER INTERRUPT. >> SO HOW DOES THIS TIE IN TO THE SUSCEPTIBILITY TO DISEASES? BECAUSE INDIVIDUALS WHO HAVE SICKLE CELL DISEASE ARE VERY SUSCEPTIBLE TO LITERALLY ANYTHING AND EVERYTHING, SO HOW CAN THIS KIND OF HELP IN A WAY? >> SO HOW CAN WE -- YOU KNOW, HAVING THIS BETTER SENGS OF WHAT IS GOING WRONG, BUT YOU'RE ABSOLUTELY RIGHT, IN INDIVIDUALS LIVING WITH THE DISEASE, EVEN THOUGH IT'S A RED CELL DEFECT, IT'S AN INFLAMMATORY STATE, THE WHITE CELLS SEEM TO BE TICKLED ALL THE TIME, CYTOKINES ARE INCREASED AND PRODUCE -- LIKE IF YOU LOOK AT SOME OF THESE MARKERS IN THE BLOOD OF SICKLE CELL INDIVIDUALS, THEY'RE UP. AND SO TYING IN WHAT'S CAUSING WHAT, EVEN IN SOME OF THE AREAS THAT WE'RE LOOKING AT FROM AN ENGINEERING POINT OF VIEW, WE'RE SAYING WHAT IS THE PRIMARY FACTOR UNDER WHAT CONDITIONS. SO EVEN BEING ABLE TO TEASE THAT OUT, I THINK IS A BIG ISSUE. SO I DON'T KNOW THAT WE HAVE AN EASY ANSWER TO THAT RIGHT NOW, BUT THAT'S SOMETHING THAT WE SHOULD BE WORKING ON AS WELL. AND ANOTHER THING THAT I'LL JUST SAY QUICKLY THAT WE'RE TRYING TO DO FROM AN ENGINEERING POINT OF VIEW, TO TAKE A SYSTEMS APPROACH TO SAY THAT YOU HAVE MULTI-ORGAN DAMAGE, YOU HAVE MULTIPLE CELL TYPES INVOLVED, DIFFERENT -- VARIABILITY AMONGST PATIENTS, VARIABILITY IN PATIENTS, IF WE COULD JUST START GRABBING ALL THAT DATA, PUT IT TOGETHER AND THEN MAYBE MINE IT IN A SYSTEMATIC WAY, THAT MIGHT ALSO GIVE US SOME POINTERS ON DIRECTIONS TO MOVE INTO TO MAKE SOME CHANGES. >> THANK YOU. >> THANK YOU FOR YOUR INSIGHTS. >> -- NNIDDK. THANK YOU VERY MUCH FOR YOUR PRESENTATION. MY QUESTION IS, DID YOU SEE A REVERSAL OF THE FLOW WITHIN THE MICRO -- THE SIMULATED -- THE VIDEO YOU SHOWED WITH THE MICRO VESSELS, WAS IT WITH IRREVERSIBLE SICKLE CELLS THAT HYDROXYUREA HAD THAT EFFECT? IF YES, DOES IT SUPPOSE THAT IT AFFECTS THE SHAPE IN SOME WAY AND NOT JUST THE POLYMERIZATION OF THE HEMOGLOBIN? >> SO IN OUR ASSAYS, THEY WEREN'T DONE UNDER DEOXYGENATED CONDITIONS, SO WE HAD THE SAMPLE AND IT IS WHAT IT IS. SO THE SHAPES ARE -- THE CELLS ARE IN WHATEVER SHAPE THAT HE ARE AT THAT TIME. AND THEN WE'RE FLOWING THEM IN A ONE-WAY FLOW SYSTEM. SO WE'RE NOT -- IN THAT SYSTEM, WE'RE NOT SEEING REVERSIBILITY BECAUSE WE WEREN'T CONTROLLING FOR THE OXYGENATION. BUT IF YOU HAVE A SYSTEM WHERE YOU ARE CONTROLLING FOR THE OXYGENATION, IT'S POSSIBLE THAT YOU MIGHT OBSERVE SOME REVERSIBLE SHAPE CHANGE IN THAT OXYGENATED VERSUS DEOXYGENATED ENVIRONMENT. THE PROBLEM WE HAVE WAS THAT IF YOU'RE TRYING TO HAVE ENDOTHELIAL CELLS AND THE RED CELLS PRESENT AND MICROSCOPY SO YOU CAN SEE IT ALL, IT'S HARD TO DO IT ALL IN ONE SYSTEM, SO THE PEOPLE WHO TEND TO HAVE DONE SOME DEOXYGENATED/OXYGENATED STUDIES HAD CHANNELS THAT WERE NOT END ENDOTHEELIZED FLOWING THROUGH THIS CHANNEL THAT YOU HAVE OXYGEN BUT YOU DON'T HAVE ENDOTHELIAL CELLS PRESENT. IN OURS, WE HAD ENDOTHELIAL CELLS BUT WE WEREN'T -- YOU'RE ENDOTHELIAL CELLS ARE NOT REMAINING VIABLE, SO THAT'S WHAT I MEAN TOO BY HAVING MORE OF A SYSTEMIC APPROACH OR SYSTEMATIC APPROACH WHERE YOU COULD ACTUALLY SAY I HAVE A SET OF DATA FROM THIS TYPE OF SYSTEM, A SET OF DATA FROM THIS TYPE OF SYSTEM, AND ONE OF THE REASONS WHY WE'RE COMBINING IN VITRO WITH THE ANIMAL STUDIES IS BECAUSE ANIMAL STUDIES AT LEAST GIVE YOU INTACT VESSELS AND YOU HAVE THE OXYGENATED SYSTEM, IT'S ALL INTACT. BUM THEY'RE ANIMALS, THEY'RE NOT HUMANS. AND THE IN VITRO SYSTEM WILL ALLOW US TO USE HUMAN CELLS, BUT IT'S NOT INTACT. SO IF WE CAN COMBINE THE DATA AND HAVE THAT ONE SET CORROBORATE ON ANOTHER SET OR GIVE INSIGHTS INTO WHAT YOU FOUND IN ONE ASSAY SYSTEM, HOPEFULLY PUTTING THAT ALL TOGETHER WILL GIVE US A BETTER PICTURE. >> ALL RIGHT. THANK YOU VERY MUCH. MY SECOND SMALL OBSERVATION WHICH IS A BIT SCARY AS WELL BECAUSE IN THE SICKLE CELL TRAIT, WE SEE THE HUGE BONE CHANGES, BUT CLINICALLY, THEY DON'T SEEM TO HAVE THE SAME TYPE OF BONE PAINS THAT SICKLE CELL PATIENTS HAVE. WHY DO YOU THINK WE HAVE THAT KIND OF BONE CHANGE IN THE TRAIT? >> SO HAVING ANY SICKLE HEMOGLOBIN PRESENT AT ALL CAN ACTUALLY CAUSE SOME KINDS OF IMPLICATIONS. BUT -- BECAUSE ALSO -- BUT HAVING LARGE PERCENTAGES OF NON-SICKLED HEMOGLOBIN IS PROTECTIVE. SO IN THE TRAIT, THAT'S PRETTY MUCH FOR THE MOST PART PROTECTIVE AGAINST THE KINDS OF DAMAGE THAT YOU WOULD SEE IF YOU'RE HOMOZYGOUS. SO THAT'S PART OF THE EXPLANATION. BECAUSE ALSO, IF YOU'RE HOMOZYGOUS AND YOU'RE HAVING THESE CONSTANT OCCLUSIONS AND CRISES AND CONSTANT INFLAMMATION, YOU'RE GOING TO SEE A LOT MORE THAN YOU WOULD SEE OTHERWISE. SO THAT'S PART OF THE REASON WHY WE WERE SAYING WE NEED TO DO THINGS TO LOOK MORE AT THE MICRO ARCHITECTURE, BECAUSE MAYBE ON THE SURFACE, ON MACRO MEASUREMENTS, THE TRAIT SEEMS LIKE IT'S OKAY, BUT THERE MIGHT BE SOME MICRO-TYPE BEHAVIOR THAT YOU WOULDN'T PICK UP. BUT OVERALL, INDIVIDUALS WITH TRAIT DO SEEM TO BE LARGELY ASYMPTOMATIC EXCEPT UNDER CERTAIN CONDITIONS. >> THANK YOU VERY MUCH. >> SURE. >> NOT SEEING ADDITIONAL QUESTIONS, I WAS ASKED BY DR. TABAK -- OBVIOUSLY I'M NOT DR. TABAK -- TO REBLIND YOU THAT REMIND YO U THAT THERE'S GOING TO BE A RECEPTION SPONSORED BY FAES RIGHT OUTSIDE OF THE AUDITORIUM HERE, AN WE'RE GOING TO ASK YOU TO AGAIN THANK DR. BARABINO FOR A LOVELY TALK. >> THANK YOU.