Buck Institute For Research On Aging

REVERSIBLE MITOCHONDRIAL PROTEIN ACETYLATION AND METABOLIC REGULATION

CELLULAR SENESCENCE AND CELL FATE/INTERACTIONS AS DRIVERS OF ALZHEIMER'S AND AGE-RELATED DEMENTIAS - OVERALL PROJECT SUMMARY AGING IS BY FAR THE MOST IMPORTANT DRIVER AND RISK FACTOR FOR DEVELOPING A VARIETY OF NEURODEGENERATIVE DISEASES, INCLUDING ALZHEIMER’S DISEASE (AD) AND RELATED DEMENTIAS. THESE DEVASTATING DISEASES EXACT AN ENORMOUS EMOTIONAL, SOCIAL AND ECONOMIC TOLL ON PATIENTS AND THEIR FAMILIES, YET TO DATE THERE ARE NO EFFECTIVE TREATMENTS THAT DELAY, MUCH LESS REVERSE, THE ONSET OR PROGRESSION OF THESE DISEASES. CLEARLY, NEW APPROACHES TO UNDERSTANDING AND TREATING AGE-RELATED NEURODEGENERATION ARE NEEDED. THIS PROGRAM PROJECT GRANT (PPG) PROPOSAL AIMS TO FILL THIS SERIOUS GAP IN OUR KNOWLEDGE AND TREATMENT APPROACHES. THE PROPOSED PPG CONSISTS OF THREE RESEARCH PROJECTS, EACH FOCUSED ON AN ASPECT OF BRAIN AGING THAT IS KNOWN TO BE CRUCIAL FOR BRAIN FUNCTION: 1) CELL FATE DECISIONS, PARTICULARLY CELL DEATH AND CELLULAR SENESCENCE; 2) METABOLISM, PARTICULARLY RESPONSES LEADING TO METABOLIC REPROGRAMMING AND INFLAMMATION; AND 3) CELL-CELL INTERACTIONS, PARTICULARLY INTERACTIONS BETWEEN NEURONS AND NON-NEURONAL CELLS IN THE BRAIN. WE PROPOSE TO SUPPORT THE PROJECTS BY AN ADMINISTRATIVE CORE, WHICH WILL ALSO PROVIDE STATISTICAL AND BIOINFORMATICS SUPPORT, AND THREE SCIENTIFIC CORES: 1) AN IPSC/ORGANOID CORE; 2) A PROTEOMICS AND METABOLISM CORE; AND 3) A SINGLE CELL AND SPATIAL TRANSCRIPTOMICS CORE. THE PPG BENEFITS FROM THE EXCEPTIONALLY DIVERSE EXPERTISE OF THE PROJECT AND CORE LEADERS AND CO-LEADERS, ALL OF WHOM ARE ACKNOWLEDGED LEADERS IN CONTEMPORARY AGING RESEARCH. EACH OF THE PROJECTS IS A CLOSE COLLABORATION AMONG SEVERAL PPG MEMBERS, MANY OF WHOM HAVE A HISTORY OF PRODUCTIVE COLLABORATION. EACH OF THE SCIENTIFIC CORES WILL PROVIDE STATE-OF-THE ART SUPPORT TO THE PROJECTS, ENABLING CONCEPTUAL AND TECHNICAL ADVANCES THAT WOULD BE DIFFICULT TO ACHIEVE IN ISOLATION. TOGETHER, THE PROJECTS AND CORES HAVE THE POTENTIAL TO UNCOVER NEW MECHANISMS OF AD AND RELATED DEMENTIAS, WHICH WILL BE TESTED IN HUMAN CELLS AND ORGANOIDS AND MICE. IMPORTANTLY, THESE MECHANISMS CAN BE DEVELOPED INTO INTERVENTIONS THAT CAN BE USED TREAT HUMAN PATIENTS.

SENESCENT CELL MAPPING, IDENTIFICATION AND VALIDATION FOR HUMAN SOMATIC AND REPRODUCTIVE TISSUES - OVERALL TMC - PROJECT SUMMARY CELLULAR SENESCENCE IS A MULTI-FACETED CELL FATE THAT ARRESTS CELL PROLIFERATION AND ACTIVATES THE SYNTHESIS AND SECRETION OF NUMEROUS CYTOKINES, CHEMOKINES, GROWTH FACTORS, PROTEASES AND LIPIDS, TERMED THE SENESCENCE ASSOCIATED SECRETORY PHENOTYPE (SASP). THE SASP CAN INFLUENCE TISSUE MICROENVIRONMENTS, LOCALLY AND DISTALLY, AND THUS SENESCENT CELLS CAN STRONGLY AFFECT TISSUE FUNCTION. SENESCENT CELLS (SNCS) INCREASE WITH AGE IN MOST VERTEBRATE ORGANISMS, INCLUDING MICE AND HUMANS, AND IT IS INCREASINGLY CLEAR THROUGH BOTH GENETIC AND PHARMACOLOGICAL MANIPULATIONS THAT THEY CAN DRIVE A GROWING LIST OF AGE-RELATED PATHOLOGIES, RANGING FROM NEURODEGENERATION TO CANCER. AT PRESENT, THERE ARE NO INVARIANT BIOMARKERS OF SNCS AND THE MOLECULAR CHARACTERISTICS OF SNCS ARE REMARKABLY HETEROGENEOUS AND VARIABLE, DEPENDING ON CELL AND TISSUE TYPE, MICROENVIRONMENT, SENESCENCE INDUCER, AND TIMING. OUR OVERALL GOAL IS TO DETERMINE, MOLECULARLY AND SPATIALLY, WHEN AND WHERE SENESCENT CELLS OCCUR IN HUMANS, AND ALSO HOW THEIR PATTERNS OF GENE EXPRESSION AND SASPS VARY WITH TISSUE PHYSIOLOGY AND AGE. THESE GOALS ARE ALSO THE GOALS OF THE SENNET CONSORTIUM. WE THEREFORE PROPOSE TO ESTABLISH A SENNET TISSUE MAPPING CENTER (TMC) COMPRISED OF AN ADMINISTRATIVE CORE, BIOSPECIMEN CORE, BIOLOGICAL ANALYSIS CORE AND DATA ANALYSIS CORE. OUR PROPOSED TMC WILL FOCUS ON THREE HUMAN TISSUES (OVARY, BREAST AND SKELETAL MUSCLE) AND THREE BIOFLUIDS (FOLLICULAR FLUID, PLASMA AND URINE). THESE SAMPLES HAVE DISTINCT BIOLOGICAL CHARACTERISTICS AND CELL TYPES (SOMATIC AND REPRODUCTIVE; STROMAL, EPITHELIAL AND VASCULATURE) THAT SHOW SIGNIFICANT CHANGES WITH AGE. ALL MATERIALS ARE AVAILABLE THROUGH ESTABLISHED COLLABORATIONS, SUBCONTRACTORS AND/OR BIOBANKS (THROUGH THE BIOSPECIMEN CORE). THEIR ANALYSES WILL INCLUDE CUTTING-EDGE TRANSCRIPTOMIC AND PROTEOMIC TECHNIQUES THAT WILL TAKE ADVANTAGE OF THE EXPERTISE OF SEVERAL BUCK INSTITUTE INVESTIGATORS. OUR PRELIMINARY DATA SHOW THAT THESE TISSUES AND BIOFLUIDS ARE AMENABLE TO THE STATE-OF-THE-ART TECHNOLOGIES PROPOSED IN THE BIOLOGICAL ANALYSIS AND DATA ANALYSIS CORES, AS WELL AS NEW TECHNOLOGIES PROPOSED IN OUR ACCOMPANYING TECHNOLOGY DEVELOPMENT APPLICATION (RFA-RM- 21-009). IMPORTANTLY, OUR FINDINGS WILL BE APPLICABLE TO MANY OTHER HUMAN TISSUES AND BIOFLUIDS IN ORDER TO ENCOMPASS AND COMPLEMENT THE OVERALL GOALS OF THE LARGER SENNET CONSORTIUM. THE ADMINISTRATIVE CORE WILL COORDINATE ALL ASPECTS OF THE TMC, FROM SPECIMEN ACQUISITION TO ANALYSIS, AND WILL OVERSEE AND FACILITATE FREQUENT INTERACTIONS BETWEEN THE PROPOSED TMC AND OTHER INVESTIGATORS, CORES AND COMPONENTS OF THE SENNET CONSORTIUM.

TRAINING IN BASIC AGING RESEARCH AND AGE-RELATED DISEASE

DISCOVERING COMPOUNDS WITH ROBUST PRO-LONGEVITY ACTIVITIES

MITOCHONDRIAL DYSFUNCTION IN AGING AND DISEASE

MOLECULAR MECHANISMS OF HIV LATENCY

AUTOPHAGY AND DIETARY RESTRICTION MECHANISMS IN THE C. ELEGANS MODEL OF AGING

IDENTIFYING FACTORS REGULATING MEDIUM SPINY NEURON DIFFERENTIATION OR MAINTENANCE AS THERAPEUTIC TARGETS FOR HUNTINGTON'S DISEASE USING INDUCED PLURI

PHARMACOLOGY OF LIFESPAN EXTENSION

THE UNIVERSITY OF SOUTHERN CALIFORNIA AND BUCK INSTITUTE NATHAN SHOCK CENTER

ROLE OF CELLULAR SENESCENCE IN CARDIOVASCULAR AGING

RETROGRADE REGULATION OF SYNAPTIC STRENGTH BY TRANSLATIONAL MECHANISMS

CELLULAR SENESCENCE AND CONTROL OF GENE EXPRESSION

RESILIENCE PATHWAYS MODELING HUMAN LONGEVITY-PROMOTING APOE VARIANTS IN INDUCED PLURIPOTENT STEM CELLS

NOVEL MODEL FOR HIV LATENCY IN ORAL PRIMARY LYMPHOID TISSUES

GEROSCIENCE METABOLITES BETA-HYDROXYBUTYRATE AND NAD+ LINKING INFLAMMATION AND NEUROENERGETIC FAILURE IN DELIRIUM

INTERDISCIPLINARY RESEARCH CONSORTIUM IN GEROSCIENCE

MECHANISMS OF THE SIGNALING METABOLITE ?-HYDROXYBUTYRATE IN ALZHEIMER'S DISEASE AND THE AGING BRAIN

QUANTITATIVE PROTEOMICS TO DEVELOP ROBUST SENESCENCE-RELATED BIOMARKERS FOR AGING

IMPAIRED ACTIVITY-DEPENDENT PROTEIN SYNTHESIS IN DENDRITES AND PATHOPHYSIOLOGY IN TAUOPATHY - PROJECT SUMMARY/ABSTRACT PATHOGENIC TAU IN ALZHEIMER’S DISEASE AND RELATED DEMENTIAS INHIBITS THE PLASTICITY OF NEURONAL CONNECTIONS UNDERLYING PROGRESSIVE MEMORY LOSS. WHAT REMAINS UNDETERMINED, HOWEVER, ARE THE COMPLEX AND MULTIFACTORIAL MOLECULAR EVENTS THAT DISRUPT SYNAPTIC PLASTICITY IN NEURONS WITH PATHOGENIC TAU. THIS IS MAJOR GAP IN OUR KNOWLEDGE OF THE MOLECULAR AND PHYSIOLOGICAL PROCESSES UNDERLYING MEMORY LOSS IN ALZHEIMER’S DISEASE AND RELATED DEMENTIAS. THERE IS A CRITICAL NEED TO UNCOVER HOW PLASTICITY IS DYSREGULATED BY PATHOGENIC TAU TO LAY THE GROUNDWORK FOR NEW STRATEGIES TO TARGET PLASTICITY FOR THERAPEUTIC INTERVENTION IN THESE DISEASES. LONG-TERM POTENTIATION (LTP) IS A TYPE OF PLASTICITY THAT INVOLVES THE PERSISTENT STRENGTHENING OF SPECIFIC CONNECTIONS BETWEEN NEURONS IN RESPONSE TO INCREASED ACTIVITY. THERE IS A STRONG LINK BETWEEN MEMORY IMPAIRMENTS AND OBSTRUCTED LTP AT SYNAPSES IN TRANSGENIC MOUSE MODELS OF ALZHEIMER’S DISEASE AND RELATED DEMENTIAS, SUGGESTING THAT THE INABILITY OF SYNAPSES TO EXPRESS LTP IS A KEY FACTOR LEADING TO MEMORY DECLINE. OUR PRELIMINARY STUDIES SUGGEST THAT PATHOGENIC TAU BLOCKS THE ACTIVITY-DEPENDENT DE NOVO PROTEIN SYNTHESIS IN DENDRITES THAT IS REQUIRED FOR LTP EXPRESSION. MOREOVER, WE FOUND THAT PATHOGENIC TAU DOWNREGULATES A TRANSLATION INITIATION FACTOR THAT CONTROLS ACTIVITY-DEPENDENT MRNA TRANSLATION DURING PLASTICITY. THE CENTRAL HYPOTHESIS OF THIS PROPOSAL IS THAT PATHOGENIC TAU BLOCKS THE INITIATION OF LOCAL PROTEIN TRANSLATION IN DENDRITES AND THEREBY DISRUPTS THE DYNAMICS OF POSTSYNAPTIC PROTEINS THAT ESTABLISH LTP AND THE ENCODING OF NEW MEMORIES. TO DELINEATE THE MECHANISM BY WHICH PATHOGENIC TAU INHIBITS LTP EXPRESSION, WE PROPOSE TO USE BOTH HUMAN INDUCED PLURIPOTENT STEM CELL (IPSC)-DERIVED CULTURED NEURONS WITH FAMILIAL TAU MUTATIONS, V337M AND R406W, THAT CAUSE DEMENTIA AND THE PS19 TAUOPATHY MOUSE MODEL. WE WILL ESTABLISH THE EFFECT OF PATHOGENIC TAU ON THE ACTIVE TRANSLATION OF MRNAS INTO NEWLY SYNTHESIZED PROTEINS DURING LTP EXPRESSION. WE WILL USE A COMBINATION OF METHODS TO MONITOR THE EFFECT OF TAU ON THE RATE OF PROTEIN SYNTHESIS AND ON THE SUBSET OF MRNAS THAT ARE ACTIVELY TRANSLATED DURING LTP. WE WILL NEXT DETERMINE THE IMPACT OF PATHOGENIC TAU ON A CRITICAL STEP IN THE INITIATION OF PROTEIN TRANSLATION AND THE EXTENT TO WHICH RESTORING ACTIVITY-DEPENDENT TRANSLATION INITIATION CAN AMELIORATE LTP AND MEMORY IMPAIRMENTS IN PS19 MICE ASSESSED BY ELECTROPHYSIOLOGICAL RECORDINGS AND BEHAVIORAL TESTS OF LEARNING AND MEMORY. FINALLY, WE WILL USE AN UNBIASED AND INNOVATIVE APEX PROTEOMICS APPROACH TO ESTABLISH THE IMPACT OF PATHOGENIC TAU ON THE DYNAMIC CHANGES IN THE POSTSYNAPTIC PROTEIN COMPOSITION DURING LTP EXPRESSION THAT INVOLVE BOTH DE NOVO PROTEIN SYNTHESIS AND THE REORGANIZATION OF PROTEIN COMPOSITION TO ENHANCE SYNAPTIC STRENGTH. FROM THESE STUDIES, WE EXPECT TO ELUCIDATE THE EFFECT OF PATHOGENIC TAU ON DENDRITIC PROTEIN SYNTHESIS AND THE REGULATION OF POSTSYNAPTIC PROTEIN DYNAMICS THAT UNDERLIE THE COMPLEX AND MULTIFACTORIAL CAUSES OF MEMORY LOSS IN ALZHEIMER’S DISEASE AND RELATED DEMENTIAS.

NEW MECHANISTIC INSIGHTS INTO HOW THE GUT METABOLITE UROLITHIN A EXTENDS LIFESPAN AND PREVENTS AD

NUTRIENT SIGNALING AND STEM CELL MAINTENANCE IN AGING EPITHELIA

MASS SPECTROMETRY AND IMAGING TECHNOLOGIES IN GEROSCIENCE

IDENTIFICATION OF HIV LATENCY BIOMARKERS WITH A DUAL FLUORESCENCE REPORTER HIV

ADVANCED GLYCATION ENDPRODUCTS (AGES) AS METABOLIC BY-PRODUCTS THAT MEDIATE NEURODEGENERATION.

PROTEOSTASIS AND METABOLISM IN BRAIN AGING

NEURONAL FXR AS A POTENTIAL THERAPEUTIC TARGET FOR ALZHEIMER'S DISEASE

A TEMPORAL BIOENERGETIC, METABOLOMICS, AND PROTEOMIC MAP OF ALZHEIMER'S DISEASE IN INVERTEBRATE MODELS

SYSTEM APPROACHES TO DETERMINE MECHANISMS UNDERLYING YEAST REPLICATIVE AGING

TARGETING AGING TO PREVENT ALZHEIMER'S DISEASE: THE GEROSCIENCE APPROACH

THE B CELL INSULIN RECEPTOR IN HEALTH AND IN INSULIN RESISTANCE - PROJECT SUMMARY / ABSTRACT OBESITY IS A MAJOR GLOBAL HEALTH CONCERN. WHEN PEOPLE BECOME OBESE, THE BODY FAILS TO RESPOND WELL TO INSULIN, CALLED INSULIN RESISTANCE. THIS PROCESS CAN LEAD TO HIGH BLOOD SUGAR TRIGGERING TYPE 2 DIABETES, THOUGH THE UNDERLYING CAUSES ARE POORLY UNDERSTOOD. WE HAVE SHOWN THAT INFLAMMATION IN THE LIVER AND FAT ARE MAJOR CAUSES OF INSULIN RESISTANCE. FAT TISSUE IN MICE AND PEOPLE HAVE INCREASED IMMUNE CELLS, T AND B CELLS, THAT CAUSE INFLAMMATION. THIS NET INFLAMMATION IS ONE KEY LINK LEADING TO OBESITY RELATED INSULIN RESISTANCE. THE CURRENT RESEARCH PROPOSAL INVESTIGATES HOW THE B CELL BEHAVES DURING EARLY AND LATER STAGES OF OBESITY. INTERESTINGLY, HERE WE DESCRIBE INSULIN ITSELF AS A MAJOR FACTOR DICTATING THE BEHAVIOR OF B CELLS IN OBESITY. SPECIFICALLY, WE SHOW THAT INSULIN BINDING TO ITS RECEPTOR ON B CELLS CAUSES THE B CELLS TO PROLIFERATE, MAKE INFLAMMATORY PROTEINS AND ANTIBODIES. THIS PROCESS CONTRIBUTES TO ESTABLISHMENT OF INSULIN RESISTANCE SINCE WHEN MICE ARE GENETICALLY ENGINEERED TO CONTAIN B CELLS LACKING INSULIN RECEPTORS, THE MICE SHOW IMPROVED BLOOD SUGAR LEVELS WHEN FED A DIABETES INDUCING HIGH FAT DIET FOR A LIMITED TIME. HOWEVER, THIS SAME PATHWAY MAY ALSO LIMIT IMMUNE CELL FUNCTION DURING LONGSTANDING OBESITY. INDEED, WE SEE INSULIN RESISTANCE INSIDE IMMUNE CELLS WITH LONGER DURATION HIGH FAT DIET, AND COMPROMISED RESPONSE TO VIRAL LUNG INFECTION IN MICE WITH INSULIN RESISTANT IMMUNE SYSTEMS. THUS, WE BELIEVE THAT INSULIN IS ONE CRITICAL FACTOR WHICH PRIMES THE IMMUNE SYSTEM TO RESPOND TO DANGER SIGNALS IN THE ENVIRONMENT AND FUEL ITS FUNCTION. DURING ESTABLISHMENT OF OBESITY AND INSULIN RESISTANCE, INSULIN BOOST ACTIVATION AND METABOLISM OF IMMUNE CELLS TO HEIGHTEN INFLAMMATION WHEN RESPONDING TO DANGER SIGNALS; HOWEVER, AS THE PATHWAY BECOMES RESISTANT, THESE IMMUNE CELLS WITH HIGH BASAL INFLAMMATORY TONE ARE CRIPPLED TO RESPOND TO NEW CHALLENGE SUCH AS VIRUS. THIS MECHANISM ALSO LIKELY EXPLAINS IN PART WHY THE OBESE CANNOT FIGHT OFF VIRUSES LIKE INFLUENZA OR SARS-2 CORONAVIRUSES. IN THIS PROPOSAL WE WILL USE GENETICALLY ENGINEERED MOUSE MODELS TO MAP OUT HOW INSULIN CONTROLS B CELL IMMUNOLOGY DURING DIFFERENT DURATIONS OF OBESOGENIC DIET. FIRST, WE WILL LOOK AT INSULIN’S CAPACITY TO CONTROL B CELL INFLAMMATION, METABOLIC PROGRAMMING AND ANTIBODY PRODUCTION. NEXT, WE WILL UNDERSTAND HOW DIET INDUCED OBESITY COMMUNICATES VIA INSULIN ACTION ON B CELLS TO CONTROL BLOOD SUGAR. THIS AIM INCLUDES MAPPING OUT INSULIN RECEPTOR DOCKING SITES ON IMMUNOLOGICAL TARGET GENES. THEN WE WILL CHARACTERIZE THE IMMUNOLOGICAL CONSEQUENCES OF IMMUNE CELL INSULIN RESISTANCE DURING LUNG VIRUS INFECTION IN A MOUSE MODEL OF INFLUENZA. FINALLY, WE WILL DETERMINE THE RELATIVE MECHANISTIC ROLES FOR OBESITY RELATED DANGER PATTERN SIGNALING IN CONTRIBUTING TO THE INSULIN RESISTANT B CELL INFLAMMATORY STATE. THESE EXPERIMENTS WILL GIVE CRUCIAL NEW INSIGHTS INTO IMMUNE CELL INFLUENCE ON OBESITY, AND HOW OBESITY POTENTIALLY CRIPPLES IMMUNITY, WHICH HAS RELEVANCE TO MANY CONDITIONS, INCLUDING LETHAL VIRUSES SUCH AS OUR CURRENT PANDEMIC.

MULTIDIMENSIONAL MAPPING OF PROTEOME CHANGES AND MECHANISMS UNDERLYING YEAST REPLICATIVE AGING - PROJECT SUMMARY ONE OF THE MAJOR GOALS OF AGING RESEARCH IS TO UNDERSTAND HOW CELLS GRADUALLY DEGENERATE OVERTIME. DECADES OF WORK BY MOLECULAR AND CELL BIOLOGICALS HAVE DISCOVERED SEVERAL CONSERVED HALLMARKS OF CELLULAR AGING, INCLUDING MITOCHONDRIAL DYSFUNCTION AND VACUOLE/LYSOSOME DEFECTS. THESE AGING HALLMARKS ARE USUALLY STUDIED ONE AT A TIME AND LEFT THE CAUSES AND CONNECTIONS BETWEEN THEM LARGELY UNKNOWN. WHAT ARE THE TRIGGERS AND CONNECTIONS BETWEEN MOLECULAR EVENTS THAT STEP-BY-STEP CULMINATE AT THE DIFFERENT HALLMARKS OF AGING? WITHOUT THE BIG PICTURE OF HOW EACH PROTEIN CHANGES IN A PROTEOME, OFTEN THE BEST WE CAN DO IS GUESS AND TRY WHEN IT COMES TO THE CAUSES AND CONNECTIONS BETWEEN THESE HALLMARKS. TO FILL THE GAPS, WE HAVE DEVELOPED AND IMPLEMENTED A NEW HIGH-THROUGHPUT IMAGING METHOD TO SYSTEMATICALLY TRACK THE FATE OF EACH INDIVIDUAL PROTEIN, INCLUDING ITS EXPRES- SION, LOCALIZATION, AGGREGATION, AND TIMING OF THESE CHANGES (4D FATE MAP), DURING THE REPLICATIVE AGING OF BUDDING YEAST. THE ONGOING 4D FATE MAPPING EFFORT HAS DEMONSTRATED ITS VALUE IN REVEALING NOVEL AGE-RELATED MOLECULAR SIGNATURES AND THE PRIMARY CAUSES FOR THE WELL-KNOWN HALLMARKS OF AGING. A REPRESENTATIVE EXAMPLE FROM OUR PILOT FATE MAPPING EFFORT IS THAT THE AGE-ASSOCIATED REDUCTION OF TOM70 IS A KEY EVENT OF MITOCHONDRIAL AGING. OVEREXPRESSING TOM70 CAN PREVENT THE AGE-ASSOCIATED DEFECTS IN MITOCHONDRIAL BIOGENESIS AND VACUOLE ACIDI- FICATION--TWO HALLMARKS OF CELLULAR AGING. THE GOAL OF THIS PROPOSAL IS TO TAKE A DEEP DIVE INTO THE MOLECULAR MECH- ANISMS OF THESE NOVEL TOM70 FUNCTIONS, WHICH WILL SERVE AS AN EXAMPLE OF USING PROTEOME FATE MAPPING TO REVEAL UNKNOWN CAUSES AND CONNECTIONS OF AGING HALLMARKS (AIM 1 AND AIM 2). AS OUR PRELIMINARY STUDY HAS CLEARED THE TECHNICAL BARRIERS, WE WILL FINISH THE MAPPING OF ENTIRE YEAST PROTEOME WITHIN THIS GRANT CYCLE. THE COMPLETION OF FATE MAPPING (AIM 3) WILL NOT ONLY COMPLEMENT THE OTHER TWO AIMS BY UNFOLDING ADDITIONAL MECHANISMS THAT CONTRIBUTE TO THE AGING OF MITOCHONDRIA AND VACUOLE (AIM 1 AND AIM 2) BUT ALSO FILL THE GAPS BETWEEN ALL OTHER AGING HALLMARKS. THIS IS EXEMPLIFIED BY OUR PRELIMINARY RESULTS THAT THE ONGOING FATE MAPPING PROVIDED AN UNEX- PECTED MECHANISM OF TOM70 REDUCTION DURING AGING. WE EXPECT TO REVEAL ADDITIONAL MECHANISMS OF TOM70 REDUCTION AND MITOCHONDRIAL/VACUOLAR AGING WHEN FATE MAPPING COVERS MORE PROTEINS. TOGETHER, THIS PROJECT WILL ADVANCE BOTH THE DEPTH AND BREADTH OF OUR UNDERSTANDING OF AGING AND PROVIDE EXAMPLES OF HOW THE FATE MAP (AIM 3) CAN BE USED TO COMPREHENSIVELY UNDERSTAND THE MULTIFACTORIAL CAUSES OF AGING HALLMARKS (E.G., MITOCHON- DRIAL AND VACUOLE DEFECTS IN AIM 1 AND AIM 2). MINING THE 4D MAP OF PROTEOME AGING BY THE RESEARCH COMMUNITY WILL SYSTEMATICALLY UNVEIL PREVIOUS UNKNOWN MECHANISMS AND CONNECTIONS BETWEEN DIFFERENT AGING HALLMARKS. THE COMPLETION OF THIS PROJECT WILL BENEFIT RESEARCHERS SEEKING THE TRIGGERS AND CONNECTIONS BETWEEN DIFFERENT AGING HALLMARKS.

REGULATION OF UV-INDUCED APOPTOSIS

MRNA TRANSLATION, TOR AND GEROSCIENCE (2 OF 11)

CELLULAR AND CIRCUIT MECHANISMS OF NEUROPEPTIDE SIGNALING

CELLULAR SENESCENCE AS A MEDIATOR OF MITOCHONDRIAL DYSFUNCTION-INDUCED AGING

MOLECULAR MECHANISMS UNDERLYING THE PRESERVATION OF NEURAL STEM CELL QUIESCENCE DURING AGING

METHYLGLYOXAL DRIVES ASTROCYTE SENESCENCE TO MEDIATE NEURODEGENERATION IN ALZHEIMER'S DISEASE

MTORC1 SIGNALING IN AGING AND METABOLISM

ROLE OF SELECTIVE AUTOPHAGY IN ORGANISMAL HEALTH - PROJECT SUMMARY AUTOPHAGY IS A CELLULAR, HOMEOSTATIC PROCESS WITH IMPORTANT ROLES IN AGING AND AGE-RELATED DISEASES. DURING AUTOPHAGY, CYTOSOLIC MATERIAL OR CARGO IS SEQUESTERED INTO AUTOPHAGIC VESICLES CALLED AUTOPHAGOSOMES FOR SUBSEQUENT LYSOSOMAL DEGRADATION; HOWEVER, THE UNDERLYING MECHANISMS FOR HOW THE TUROVER OF SPECIFIC TYPES OF CARGO, E.G., PROTEIN AGGREGATES OR MITOCHONDRIA, COLLECTIVELY REFERRED TO AS SELECTIVE AUTOPHAGY, CONTRIBUTES TO CELLULAR PROTEOSTASIS AND ORGANISMAL HEALTH REMAIN ELUSIVE. WE AND OTHERS RECENTLY SHOWED A CONSERVED ROLE FOR THE FIRST IDENTIFIED AUTOPHAGY CARGO RECEPTOR IN METAZOANS, P62/SQSTM1 IN MEDIATING PROTEOSTASIS AND ORGANISMAL BENEFITS IN AN AUTOPHAGY-DEPENDENT MANNER. IN PARTICULAR, WE SHOWED THAT P62 OVEREXPRESSION IN THE SHORT-LIVED NEMATODE C. ELEGANS IS SUFFICIENT TO INDUCE AUTOPHAGY AND EXTEND LIFE- AND HEALTHSPAN, AND PROTECT AGAINST PROTEIN AGGREGATION PREDOMINANTLY IN NEURONS. MOREOVER, P62 IS SELECTIVELY REQUIRED FOR THE BENEFICIAL EFFECTS OF A HORMETIC HEAT SHOCK, A CONSERVED LONGEVITY REGIMEN THAT WE PREVIOUSLY SHOWED TO INDUCE AUTOPHAGY. COLLECTIVELY, THESE STUDIES SUGGEST THE HYPOTHESIS THAT P62 PLAYS AN INSTRUCTIVE ROLE IN INDUCING TISSUE-SPECIFIC, SELECTIVE AUTOPHAGY TO FACILITATE ORGANISMAL BENEFITS. THIS IS A NOVEL CONCEPT BECAUSE AUTOPHAGY RECEPTORS ARE TRADITIONALLY VIEWED AS FACTORS FACILITATING CARGO SEQUESTRATION, RATHER THAN BEING ACTIVE INDUCERS OF THIS COMPLEX, MULTI-STEP TURNOVER PROCESS. WE PROPOSE TO ADDRESS THIS HYPOTHESIS BY USING C. ELEGANS TO INVESTIGATE THE MOLECULAR MECHANISMS BY WHICH A HORMETIC HEAT SHOCK VIA P62 OR DIRECT P62 OVEREXPRESSION REGULATE AUTOPHAGY CELL AUTONOMOUSLY AND IMPROVE ORGANISMAL HEALTH VIA CELL NON-AUTONOMOUS EFFECTS. SPECIFICALLY, IN AIM 1, WE WILL USE GENETIC APPROACHES TO TEST HOW A HORMETIC HEAT SHOCK ENGAGES P62 TO REGULATE AUTOPHAGY ESPECIALLY IN NEURONS. IN AIM 2, WE WILL INVESTIGATE THE CELL-AUTONOMOUS AND CELL NON-AUTONOMOUS MECHANISMS BY WHICH P62 INDUCE AUTOPHAGY AND IMPROVE FITNESS. FINALLY, IN AIM 3, WE WILL USE GENETIC AND BIOCHEMICAL APPROACHES TO IDENTIFY NEW P62-RELEVANT CARGO AND P62-LIKE RECEPTORS ENGAGED BY HORMETIC HEAT SHOCK. THESE STUDIES ARE SIGNIFICANT BECAUSE THEY WILL ADVANCE OUR KNOWLEDGE OF HOW SELECTIVE AUTOPHAGY CONTRIBUTES TO ORGANISMAL HEALTHSPAN. THESE STUDIES ARE INNOVATIVE BECAUSE THEY USE A POWERFUL COMBINATION OF TECHNIQUES AND MODELS TO INVESTIGATE THE NOVEL REGULATORY CONCEPT IN THE AUTOPHAGY FIELD THAT AN AUTOPHAGY RECEPTOR IS SUFFICIENT TO INDUCE BENEFICIAL AUTOPHAGY, POTENTIALLY VIA CELL NON-AUTONOMOUS MECHANISMS. SINCE AUTOPHAGY PLAYS CRITICAL ROLES IN MANY HUMAN AGE-RELATED DISORDERS, UNDERSTANDING THE REGULATION OF AUTOPHAGY AND THE CONSERVED MECHANISMS BY WHICH AUTOPHAGY AFFECT AGING IN MULTICELLULAR ORGANISMS LIKE C. ELEGANS ARE LIKELY TO PROVIDE NEW IMPORTANT INSIGHTS RELEVANT TO AGING, WHICH ALSO MAY HELP DEVELOP TREATMENTS FOR AGE-RELATED DISEASES.

VEGF IN NEURPROTECTION AND NEUROGENESIS

PIONEER TRANSCRIPTION FACTORS IN AGING AND NEURODEGENERATION - PROJECT SUMMARY A NUMBER OF TRANSCRIPTIONAL REGULATORS HAVE BEEN FOUND TO REGULATE THE HALLMARKS OF AGING. IN PREVIOUS WORK, WE FOUND THAT THE FOXO FAMILY OF TRANSCRIPTIONAL REGULATORS, WHICH HAVE BEEN IMPLICATED IN HEALTHY AGING ACROSS SPECIES, DIRECTLY REGULATE A CONSERVED NETWORK OF TARGET GENES. IN MICE, FOXOS ARE CENTRAL REGULATORS OF STEM CELL HOMEOSTASIS DURING AGING AND ARE CRITICAL FOR TISSUE INTEGRITY. IN HUMANS, SNPS IN THE FOXO3 LOCUS HAVE BEEN LINKED TO LONGEVITY, AND THE UPSTREAM REGULATOR OF FOXO’S, INSULIN/IGF SIGNALING MUST BE TIGHTLY REGULATED TO PRESERVE HEALTHY AGING. YET, WE STILL LACK AN UNDERSTANDING OF HOW FOXOS FUNCTION AT THE CHROMATIN LEVEL, AND HOW THEIR ACTIVITY IS ALTERED DURING AGING AND IN THE CONTEXT OF NEURODEGENERATION. IN PRELIMINARY WORK, WE IDENTIFIED FOR THE FIRST TIME THE DIRECT NETWORK OF FOXO3 TARGETS IN HUMAN CELLS, AND FOUND THAT FOXOS FUNCTION AS PIONEER FACTORS TO DEPLOY A SECONDARY NETWORK OF TRANSCRIPTIONAL REGULATORS TO EXTEND THEIR TARGET GENE NETWORK. HERE, WE WILL ADDRESS THIS CRITICAL QUESTION OF THE MECHANISMS UNDERLYING THE PIONEER ACTIVITY, ITS HETEROGENEOUS NATURE, AND HOW IT CHANGES IN THE CONTEXT OF AGING AND ALZHEIMER’S DISEASE. COMPLETION OF THIS WORK WILL REVEAL THE CHROMATIN-LEVEL CHANGES THAT INFLUENCE THE ACTIVITY OF FACTORS THAT COUNTER AGING AND NEURODEGENERATION, WHICH MAY LEAD TO STRATEGIES TO IMPROVE ASSOCIATED PATHOLOGIES.

MECHANISM OF ORGANELLE DYSFUNCTION DURING AGING AND THE RELATED REJUVENATION PROCESS

A SMART CELL DRUG (SMACD) DELIVERY PLATFORM FOR MOBILE, TARGETABLE, AND SELF-REGULATED COMBINATION THERAPY: A MODEL PROJECT TO RESCUE ANTIBODIES FROM ALZHEIMER'S DISEASE (AD) CLINICAL TRIAL FAILURES - PROJECT SUMMARY / ABSTRACT AS WITH MANY OTHER DISEASES, ALZHEIMER’S DISEASE (AD) IS A MULTIFACTORIAL DISORDER CONSISTING OF MULTIPLE PATHOLOGIES THAT ARE EACH KNOWN TO BE DELETERIOUS IN THEIR OWN RIGHT. CLINICAL TRIALS ASSESSING DISEASE-MODIFYING DRUGS FOR ALZHEIMER’S DISEASE (AD) HAVE LARGELY BEEN FOCUSED ON AMYLOID BETA (ASS) PATHOLOGY RATHER THAN OTHER PATHOLOGICAL COMPONENTS INCLUDING TAU PATHOLOGY AND GLIOSIS. THIS IS IN PART DUE TO ASS PATHOLOGY BEING VIEWED AS THE INITIATING EVENT RESPONSIBLE FOR ALL OTHER PATHOLOGIES. THE INTERIM RESULT IS ADUCANUMAB (ADUHELMTM; BIOGEN) RECENTLY APPROVED BY THE FEDERAL DRUG ADMINISTRATION (FDA)L, LIKELY TO BE EFFECTIVE AT EARLY STAGES OF AD WHERE IT WOULD BE MOST EFFECTIVE IN HALTING DOWNSTREAM PATHOLOGIES. TREATMENTS FOR LATER STAGES OF DISEASES WILL REQUIRE THERAPEUTICS NOT ONLY FOR ASS, BUT ALSO TAU PATHOLOGY AND GLIOSIS NAMELY COMBINATION THERAPY (CT). CT HAS RECENTLY BEEN STRONGLY ENCOURAGED BY THE EUROPEAN UNION-NORTH AMERICAN CLINICAL TRIALS IN ALZHEIMER’S DISEASE TASK FORCE (EU/US CTAD TASK FORCE) AND THE FDA5. ADUCANUMAB ON ITS OWN IS ALREADY DOSE-LIMITED DUE TO ITS POTENTIAL FOR OFF-TARGET TOXICITY AND DEVELOPMENT OF A CT INCLUDING ITS USE WOULD RESULT IN ADDITIVE TOXICITY THAT THE OTHER DRUGS MAY INTRODUCE PLUS POSSIBLE UNKNOWN DRUG INTERACTIONS. THIS MAKES CT EXTREMELY COMPLEX TO RESEARCH AND TO DATE UNATTRACTIVE FOR BIG PHARMA. TO OVERCOME THESE CHALLENGES, WE HAVE DESIGNED A SMART CELL-BASED DRUG DELIVERY (SMACD) SYSTEM TO ENABLE IN VITRO RESEARCH HIGHLY TRANSLATABLE TO IN VIVO VALIDATION THAT HAS THE CAPABILITY OF SUPPORT A LARGE NUMBER OF DRUGS CURRENTLY BEING USED IN CLINICAL TRIALS AND TO DELIVER THEM IN COMBINATION TO THE BRAIN WITHOUT THE TOXICITY ASSOCIATED WITH SYSTEMIC OR OTHER FORMS OF BULK DRUG ADMINISTRATION. WE PROPOSE TO USE SMACD TO TARGET THE MOST AFFLICTED BRAIN REGIONS IN AD EQUIPPED WITH A CT BASED ON ADUCANUMAB AND ZAGOTENEMAB TO ELICIT MICROGLIA-MEDIATED CLEARANCE OF NEUROTOXIC OLIGOMERIC ASS AND DISEASE- SPREADING TAU, RESPECTIVELY, ACCOMPANIED BY AN IMMUNOMODULATOR TO ENHANCE THE PHAGOCYTIC ACTIVITY OF MICROGLIA WHILST SUPPRESSING INFLAMMATION. WE PLAN TO TEST THE TECHNOLOGY INITIALLY IN HUMAN CELLS AND MOUSE CELLS FOLLOWED BY IN VIVO VALIDATION IN A TRANSGENIC MOUSE MODEL OF AD THAT PRESENTS ASS AND TAU PATHOLOGY ACCOMPANIED BY GLIOSIS AT MID AND LATE STAGES OF DISEASE DEVELOPMENT. OUR RESULTS, IF SUCCESSFUL, WOULD DIRECTLY PROVIDE ‘PROOF-OF-CONCEPT’ FOR THE TRANSLATABILITY OF SMACD-DELIVERED CT DRUG THERAPY FOR USE IN THE INTERVENTION OF HUMAN DISEASE.

SYSTEMIC COORDINATION OF STRESS RESPONSES BY INSULIN SIGNALING

MOLECULAR MECHANISMS OF LIFESPAN EXTENSION BY DIETARY RESTRICTION IN DROSOPHILA

THE ROLE OF DIET AND AGING IN PHOTORECEPTOR HOMEOSTASIS AND VISUAL FUNCTION DECLINE - PROJECT SUMMARY/ABSTRACT MORE THAN HALF OF THE ELDERLY POPULATION SUFFER FROM AGE-RELATED EYE DISORDERS THAT COMPROMISE THEIR ABILITY TO PERFORM DAILY ACTIVITIES. AGING IS A SIGNIFICANT RISK FACTOR FOR SUCH DISORDERS, AND DIETARY RESTRICTION (DR) IS A ROBUST INTERVENTION THAT CAN SLOW AGING AND EXTEND HEALTHSPAN. NUTRIENT RESTRICTION INCREASES CIRCADIAN CLOCK AMPLITUDE, WHILE OVERNUTRITION AND OBESITY DAMPEN CIRCADIAN RHYTHMS, INCREASING THE RISK OF EYE DISEASES. HOWEVER, THE ROLE OF DR AND CIRCADIAN CLOCKS IN SLOWING AGE-RELATED DECLINE IN VISUAL FUNCTION IS NOT WELL UNDERSTOOD. THEREFORE, IT IS IMPERATIVE TO ADDRESS THIS GAP IN THE FIELD. THE GOAL OF THIS PROPOSAL IS TO DETERMINE THE MOLECULAR MECHANISMS THAT SLOW AGE-RELATED LOSS OF THE VISUAL SYSTEM, DEPENDING ON AGING AND DIET. WE HYPOTHESIZE THAT DR ENHANCES CIRCADIAN CLOCK GENE EXPRESSION, PROMOTING PHOTORECEPTOR HOMEOSTASIS, AND MODULATING LIFESPAN. OUR RECENT CIRCADIAN TRANSCRIPTOME PROFILING UPON DR SHOWED ENHANCEMENT OF GENES INVOLVED IN PHOTORECEPTOR HOMEOSTASIS AND DELAYED VISUAL SENESCENCE. PRIOR RESULTS SUGGEST THAT AGING LEADS TO A GRADUAL DECLINE IN CIRCADIAN RHYTHMS; HOWEVER, DR CAN SLOW THIS DECLINE. INHIBITION OF THE CIRCADIAN TRANSCRIPTION FACTOR, CLOCK (CLK), ACCELERATES THE AGE-RELATED DECLINE IN VISUAL FUNCTION, AND PHOTORECEPTOR DEGENERATION RESULTS IN SYSTEMIC INFLAMMATION AND SHORTENED LIFESPAN, WHICH DR PROTECTS AGAINST. WE HYPOTHESIZE THAT CLK PREVENTS LIGHT-INDUCED DAMAGE TO THE PHOTORECEPTORS, THUS SUPPORTING THE 'ESCAPE THE LIGHT HYPOTHESIS' FOR THE EVOLUTION OF CIRCADIAN CLOCKS. IN THIS STUDY, WE AIM TO DETERMINE THE CELLULAR PROCESSES IN PHOTORECEPTORS THAT PROTECT THEM FROM DAMAGE DUE TO AGE AND DIET, REVEALING NOVEL TARGETS FOR AGE-RELATED EYE DISEASES AND ASSOCIATED MORBIDITIES. WE PROPOSE THREE SPECIFIC AIMS TO ACHIEVE THIS GOAL. FIRST, WE WILL DETERMINE DOWNSTREAM EFFECTORS OF THE CORE CIRCADIAN CLOCK GENE, CLK, ALTERING THE AGE-RELATED DECLINE IN VISUAL SYSTEM FUNCTION UPON MODULATION OF DIET AND LIGHT. SECOND, WE WILL DETERMINE DIET-DEPENDENT TRANSCRIPTIONAL NETWORKS OF RHYTHMIC PHOTOTRANSDUCTION GENES AND THEIR IMPACT ON AGE-RELATED CHANGES IN VISUAL SYSTEM FUNCTION. THIRD, WE WILL STUDY FLY ORTHOLOGS OF GENES FROM OUR HUMAN GWAS STUDY THAT INFLUENCE RETINAL EYE AGING FOR THEIR INTERACTIONS WITH DIET, AGING, AND CIRCADIAN CLOCKS USING THE FLY. EXPECTED OUTCOMES OF THIS STUDY INCLUDE DELINEATING THE DIET AND AGING-DEPENDENT GENES INCLUDING CIRCADIAN TRANSCRIPTIONAL NETWORKS AND THEIR OUTPUTS INVOLVED IN PHOTOTRANSDUCTION HOMEOSTASIS TO MEDIATE CHANGES IN VISUAL SENESCENCE IN FLIES AND HUMANS. BY DETERMINING CELLULAR PROCESSES UNDER CIRCADIAN CONTROL IN THE PHOTORECEPTOR THAT PROTECT IT FROM DAMAGE DUE TO AGE, DIET, AND LIGHT, THIS STUDY WILL REVEAL NOVEL GENETIC TARGETS AND LIFESTYLE INTERVENTIONS MODULATING THE EFFECT OF DIET AND AGE ON EYE FUNCTION, PROVIDING POTENTIAL TREATMENT OPTIONS FOR AGE-RELATED EYE DISEASES AND ASSOCIATED MORBIDITIES.

IDENTIFYING ORGAN-SPECIFIC BLOOD BIOMARKERS OF AGING AND ASSESSING THEIR EFFECTS ON CARDIOVASCULAR HEALTH - PROJECT SUMMARY AS WE AGE, OUR ORGANS EXPERIENCE MOLECULAR AND PHYSIOLOGICAL DAMAGE THAT PREVENTS THEM FROM FUNCTIONING PROPERLY, WHICH ULTIMATELY LEADS TO DISEASE. IN THE CASE OF THE HEART, AGE-ASSOCIATED DYSFUNCTION CAUSES THE TOP KILLING AND MOST EXPENSIVE DISEASES TO TREAT WORLDWIDE. OVER THE YEARS, IT HAS BEEN ESTABLISHED THAT CARDIAC DYSFUNCTION IS LARGELY DEPENDENT ON THE INTERACTION BETWEEN THE EXPOSOME AND GENETICS; HOWEVER, ACCUMULATING EVIDENCE INDICATES THAT AGING IN DISTANT ORGANS SUCH AS THE LUNG, MUSCLE, AND KIDNEYS CAN ALSO ACCELERATE HEART AGE. IN THIS PROPOSAL, WE WILL USE DEEP LEARNING AND DEEP PHENOTYPING DATA FROM LARGE LONGITUDINAL COHORTS (BLSA, UK BIOBANK) TO GENERATE SEX-SPECIFIC CLINICAL-BASED ESTIMATORS OF THE BIOLOGICAL AGE OF DIFFERENT ORGANS AND DEFINE THE COMPLETE NETWORK OF ORGANS THAT AFFECT EACH OTHER’S AGING ACROSS POPULATIONS IN DIFFERENT SEXES. A MECHANISM THAT IS KNOWN TO DRIVE AGE-RELATED DYSFUNCTION IN THE HEART IS THE ACCUMULATION OF SENESCENT CELLS, WHICH ARE GROWTH-ARRESTED CELLS THAT SECRETE PRO-INFLAMMATORY CYTOKINES, CHEMOKINES, GROWTH FACTORS, AND PROTEASES KNOWN AS THE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP). SASP PROTEINS CAN INDUCE SENESCENCE IN HEALTHY CELLS, AND THEIR COMPONENTS HAVE BEEN SHOWN TO INCREASE WITH AGE IN THE BLOOD, SUGGESTING THAT THEY COULD MEDIATE THE PROPAGATION OF AGING BETWEEN ORGANS. IN FACT, EXPERIMENTS IN ANIMAL MODELS INDICATE THAT INDUCTION OF CELLULAR SENESCENCE IN AN ORGAN, VIA TRANSPLANTATION OR GENETICS, INDUCES CELLULAR SENESCENCE IN DISTANT ORGANS. USING BLOOD MULTI-OMICS DATA FROM INDIVIDUALS WITH CLINICAL-BASED ESTIMATIONS OF AGE IN DIFFERENT ORGANS AND NON-LINEAR MACHINE LEARNING METHODS, WE WILL GENERATE AN ATLAS OF THE SASP-LINKED BLOOD BIOMARKERS ASSOCIATED WITH BIOLOGICAL AGE IN DIFFERENT ORGANS. BY PERFORMING INTERVENTIONAL STUDIES IN HUMAN-DERIVED HEART ORGANOIDS COMBINED WITH HIGH-THROUGHPUT FUNCTIONAL ELECTROPHYSIOLOGICAL AND IMAGING ANALYSIS AND MULTI-OMICS, WE WILL STUDY THE CAUSAL EFFECTS OF SASP-ASSOCIATED PROTEINS ON CARDIAC FUNCTION. THIS WILL INDICATE THE SASP-ASSOCIATED MOLECULES SECRETED BY EACH ORGAN THAT IMPACT CARDIAC HEALTH AND ENABLE THE DISCOVERY OF NEW TARGETS TO AMELIORATE HEART-RELATED DISEASES. FINALLY, WE WILL APPLY OUR NOVEL SIGNATURE-BASED DRUG REPURPOSING METHOD (GCEA) TO IDENTIFY COMPOUNDS THAT REVERSE THE MOLECULAR CHANGES INDUCED BY MULTIPLE SASP-ASSOCIATED PROTEINS IN THE HEART. WE WILL STUDY IN MICE THE EFFECT OF THESE COMPOUNDS ON MAINTAINING CARDIAC FUNCTION DURING AGING AND THE MOLECULAR MECHANISMS INVOLVED USING MULTI-OMICS. THE EXPERIMENTALLY VALIDATED COMPOUNDS WILL HOLD GREAT POTENTIAL TO SLOW DOWN AND REVERSE THE AGING PROCESS IN THE HEART AND DELAY THE ONSET OF CARDIOVASCULAR DISEASES.

GENETIC CORRECTION OF MUTANT HUNTINGTIN IN VIVO

MOLECULAR MECHANISMS UNDERLYING THE PRESERVATION OF NEURAL STEM CELL QUIESCENCE DURING AGING - PROJECT SUMMARY COGNITIVE DECLINE IS A MAJOR FEATURE OF AGING AND NEURODEGENERATIVE DISEASES SUCH AS ALZHEIMER’S DISEASE. NEUROGENESIS, THE FORMATION OF NEW NEURONS, SUPPORTS LEARNING, MEMORY, MOOD REGULATION, AND SENSORY FUNCTIONS IN THE HEALTHY MAMMALIAN BRAIN. DURING AGING, NEUROGENESIS IS DRAMATICALLY REDUCED DUE TO IMPAIRMENTS IN NEURAL STEM CELL (NSC) POOLS. THE MECHANISMS RESPONSIBLE FOR DYSFUNCTION OF THE NSC RESERVOIR ARE NOT FULLY UNDERSTOOD, BUT SIGNIFICANT EVIDENCE IMPLICATES CHANGES TO THE QUIESCENT NSCS, WHICH ARE THE SOURCE OF THE NEUROGENIC LINEAGE. THS WORK INVESTIGATES THE MECHANISMS RESPONSIBLE FOR FAILED ACTIVATION OF QUIESCENT NSCS WITH AGE. THIS PROJECT IS SIGNIFICANT BECAUSE IDENTIFICATION OF THE MECHANISMS UNDERLYING DYSFUNCTION OF QUIESCENT NSCS WITH AGE WILL REVEAL STRATEGIES TO RESTORE NEW NEURON FORMATION IN AGING AND NEURODEGENERATION. PUBLISHED AND PRELIMINARY WORK SHOWS THAT QUIESCENT NSCS, THOUGH RELATIVELY DORMANT, ARE DEFECTIVE AT THE CHROMATIN, GENE EXPRESSION, AND METABOLIC LEVELS WITH AGE. THIS STUDY TAKES AN INTEGRATED APPROACH TO REVEAL THE SPECIFIC TARGET GENES AND PROCESSES RESPONSIBLE FOR DEFECTS IN NSC QUIESCENCE WITH AGE. SPECIFIC AIM 1 WILL INVESTIGATE HOW MITOPHAGY REGULATES METABOLIC HEALTH OF NSCS DURING AGING. AIM 2 EMPLOYS AN INTEGRATED MULTI-OMICS STRATEGY TO FULLY ELUCIDATE THE EPIGENOMIC AND METABOLIC CHANGES THAT OCCUR THE NEUROGENIC LINEAGE WITH AGE. FINALLY, AIM 3 WILL ADDRESS THE MECHANISMS BY WHICH NSCS RETURN TO QUIESCENCE, AND THE EXTENT TO WHICH FAILURE IN THIS PROCESS RESULTS IN DEPLETION OF THE FUNCTIONAL NSC POOL WITH AGE. COLLECTIVELY, THIS WORK WILL RESULT IN A COMPREHENSIVE UNDERSTANDING OF HOW DORMANT NSCS ARE AFFECTED BY AGING IN THE MAMMALIAN BRAIN. IN THE LONG TERM, THESE STUDIES HAVE THE POTENTIAL TO LEAD TO THE DEVELOPMENT OF INTERVENTIONS TO IMPROVE HEALTHY AGING AND RESTORE COGNITION IN THE CONTEXT OF NEURODEGENERATION.

HUNTINGTON'S DISEASE AND NEUROGENESIS

NEUROGLOBIN IN NEURONAL HYPOXI AND CEREBRAL ISCHEMIA

ROLE OF CIRCADIAN CLOCKS IN AGING USING DROSOPHILA

CELLULAR SENESCENCE AND ASSOCIATED LYSOSOMAL DYSFUNCTION IN IMMUNE AGING - PROJECT SUMMARY/ABSTRACT CELLULAR SENESCENCE IS AN AGE-RELATED PROCESS, AFFECTING CELLS THAT HAVE EITHER DIVIDED TOO MUCH OR SUFFERED SIGNIFICANT DAMAGE TO THEIR DNA, WHICH INVOLVES BOTH A HALT TO FURTHER PROLIFERATION AND THE ELABORATION OF A SECRETORY PROGRAM THAT ACTIVATES THE IMMUNE SYSTEM, PRESUMABLY TO FLAG THESE CELLS FOR IMMUNE CLEARANCE. ALTHOUGH THIS IS LIKELY AN IMPORTANT MECHANISM TO PREVENT CANCER, IT IS ALSO A TWO-EDGED SWORD. GROWING EVIDENCE INDICATES THAT THE ACCUMULATION OF UNCLEARED SENESCENT CELLS DRIVES A STEADY, AGE-DEPENDENT INCREASE IN BASAL INFLAMMATION, TERMED “INFLAMMAGING”, THAT CAUSES CUMULATIVE DAMAGE IN MANY TISSUES, CONTRIBUTING TO THEIR AGE-RELATED DEGENERATION. IMPORTANTLY, SENESCENT CELLS CAN INDUCE NEW SENESCENCE IN ADJACENT NORMAL CELLS VIA THEIR SECRETED FACTORS, SUGGESTING THAT THE ACCUMULATION OF SENESCENT CELLS IS A SELF- AMPLIFYING, EXPONENTIAL PROCESS. THE ROLE OF THE IMMUNE SYSTEM IN THIS VICIOUS CYCLE IS COMPLEX AND PARTICULARLY IMPORTANT, AS BASED ON AVAILABLE EVIDENCE IT BOTH MEDIATES THE INFLAMMATION-RELATED DESTRUCTION AND MAY CONTRIBUTE TO THE VICIOUS CYCLE VIA SENESCENCE WITHIN THE IMMUNE SYSTEM ITSELF. T CELLS IN THE HUMAN IMMUNE SYSTEM CAN BE VERY LONG-LIVED, AND HAVE THE ABILITY TO INFILTRATE ALMOST ANY TISSUE AND CAUSE DESTRUCTION IN VARIOUS WAYS, BUT THE ROLE OF SENESCENCE WITHIN THE T CELL REPERTOIRE IS STILL POORLY UNDERSTOOD. SEVERAL RECENTLY DEVELOPED FLOW CYTOMETRY-BASED ASSAYS CAN READILY MEASURE IN BLOOD- DERIVED IMMUNE CELLS THE LYSOSOMAL DYSFUNCTION KNOWN TO BE STRONGLY ASSOCIATED WITH CELLULAR SENESCENCE IN OTHER CELL TYPES, BUT IT IS NOT YET CLEAR IF THAT MEANS THESE CELLS ARE TRULY SENESCENT. NONETHELESS, SUCH LYSOSOMAL STRESS MAY STILL BE INDICATIVE OF POTENTIALLY DESTRUCTIVE CELLULAR PHENOTYPES, BASED ON OUR OWN PRELIMINARY DATA AND OTHER PUBLISHED STUDIES. THIS MATTERS BECAUSE THE LYSOSOMAL DYSFUNCTION, IF IT IS CAUSATIVE OF THESE DESTRUCTIVE PROPERTIES, CAN BE TARGETED BY A VARIETY OF AVAILABLE INTERVENTIONS THAT MAY PROVIDE BENEFIT REGARDLESS OF WHETHER OR NOT THE PATHOGENIC CELLS AFFECTED ARE CANONICALLY SENESCENT. CONVERSELY, IF THE CELLS IN QUESTION ARE TRULY SENESCENT, OTHER CLASSES OF DRUGS (I.E. SENOLYTICS, SENOMORPHICS) MAY INDUCE THEIR CLEARANCE OR REVERSE THEIR PATHOGENICITY. UNDERSTANDING HOW THESE CELLULAR PHENOTYPES ARISE IN THE T CELL REPERTOIRE, HOW THEY CAN BE DETECTED IN BLOOD SAMPLES, AND HOW THEY MAY BE RELATED TO IMMUNE DYSFUNCTION IN OLDER INDIVIDUALS, IS THEREFORE AN IMPORTANT UNDERTAKING FOR PUBLIC HEALTH. THIS PROPOSAL SEEKS TO ADVANCE THESE AIMS BY BRINGING TO BEAR A SUITE OF STATE-OF-THE-ART FLOW CYTOMETRY, PROTEOMIC, AND GENE EXPRESSION PROFILING-BASED TECHNIQUES, COMBINED WITH FUNCTIONAL READOUTS, TO DEFINITIVELY IDENTIFY WHICH LYSOSOMALLY STRESSED T CELLS MAY HAVE PATHOGENIC POTENTIAL, AND TO MAP THEIR OCCURRENCE ACROSS A COHORT OF 120 YOUNG AND OLD DONORS OF BOTH SEXES. THIS WORK WHEN COMPLETED WILL HAVE GENERATED A NUMBER OF NEW DIAGNOSTIC TOOLS TO DETECT SPECIFIC AGING-RELATED POPULATIONS OF T CELLS WITH POTENTIALLY HARMFUL PROPERTIES, AS WELL AS HAVING DEVELOPED SOME EARLY PRECLINICAL LEADS AS TO WHAT CLASSES OF DRUGS MAY CLEAR THEM OR MITIGATE THEIR PATHOGENICITY.

TAKEOFF: TARGETING AGING WITH KETONE ESTER IN OLDER ADULTS FOR FUNCTION IN FRAILTY - PROJECT SUMMARY/ABSTRACT THE GEROSCIENCE APPROACH OF MODULATING FUNDAMENTAL AGING MECHANISMS HOLDS GREAT PROMISE FOR GENERATING EFFECTIVE NEW THERAPEUTICS FOR COMPLEX, MULTIFACTORIAL CONDITIONS OF AGING SUCH AS FRAILTY WHICH CONTRIBUTE SO MUCH TO FUNCTIONAL DECLINE, DISABILITY, AND LOSS OF INDEPENDENCE IN OLDER ADULTS. THE FRAILTY SYNDROME IS CONCEPTUALIZED AS A CONDITION OF PROGRESSIVE FUNCTIONAL DECLINE AND INCREASED VULNERABILITY TO STRESSORS RESULTING FROM DECREASED FUNCTIONAL RESERVE. THE PATHOPHYSIOLOGY OF FRAILTY IS COMPLEX AND NOT WELL UNDERSTOOD, BUT IS GENERALLY THOUGHT TO PROMINENTLY INCLUDE CELLULAR ENERGY PRODUCTION DEFICITS ALONG WITH CHRONIC INFLAMMATION AND IMMUNE DYSFUNCTION. DIETARY RESTRICTION AND FASTING, INTERVENTIONS CLOSELY LINKED TO FUNDAMENTAL MECHANISMS OF AGING, PROMOTE THE ENDOGENOUS PRODUCTION OF KETONE BODIES AS A MEANS OF SUPPLYING FAT-DERIVED ENERGY TO TISSUES. NEW ADVANCES IN UNDERSTANDING THE BIOLOGICAL ACTIVITIES OF THE KETONE BODY BETA-HYDROXYBUTYRATE (BHB) SUGGEST THAT BOTH ENERGY PRODUCTION AND SIGNALING ACTIVITIES OF BHB MAY HAVE A MECHANISTIC ROLE IN MODULATING AGING, AND MAY BE PARTICULARLY RELEVANT TO THE PATHOPHYSIOLOGY OF FRAILTY. KETONE ESTERS HAVE RECENTLY EMERGED AS A PHARMACOLOGICAL MEANS OF DELIVERING KETONE BODIES. WE HAVE ASSEMBLED A MULTIDISCIPLINARY TEAM OF EXPERTS IN GEROSCIENCE, GERIATRICS AND FRAILTY, KETONE BODY BIOLOGY, IMMUNOSENESCENCE, AND KETONE BODY-RELATED CLINICAL TRIALS TO CARRY OUT A MULTI-SITE, PROOF-OF-CONCEPT PHASE 2A CLINICAL TRIAL OF A KETONE ESTER (KE) TARGETING FRAILTY IN PREFRAIL AND MILDLY FRAIL OLDER ADULTS. TAKEOFF IS A PROSPECTIVE, RANDOMIZED, DOUBLE-BLIND, 12-WEEK STUDY OF KETONE ESTER VS. MATCHED PLACEBO. AIM 1 WILL TEST IF MUSCLE STRENGTH AND OTHER CLINICAL FRAILTY- AND AGING-RELATED OUTCOMES IMPROVE WITH KE. AIM 2 WILL DETERMINE THE SAFETY AND TOLERABILITY OF DAILY KE IN BROADLY REPRESENTATIVE PRE-FRAIL AND FRAIL OLDER ADULTS. AIM 3 WILL TEST THE HYPOTHESIS THAT KETONE ESTER IMPACTS THE AGING IMMUNE SYSTEM, SKELETAL MUSCLE, AND BIOMARKERS OF AGING VIA DETAILED MECHANISTIC IMMUNOPHENOTYPING, MUSCLE METABOLISM, AND BIOMARKER STUDIES THAT LEVERAGE THE STRONG SPECIALIST EXPERTISE AT THE PARTICIPATING INSTITUTIONS. ALTOGETHER, THIS TRANSLATIONAL EFFORT WILL PROVIDE CRUCIAL INFORMATION ON THE SAFETY AND EFFICACY OF INTERVENTIONS INCREASINGLY BEING USED BY THE PUBLIC DESPITE A PAUCITY OF DATA, AND WITH POTENTIAL TO IMPACT IMPORTANT AGING-RELATED CONDITIONS TO IMPROVE THE HEALTH AND INDEPENDENCE OF OLDER ADULTS.

NOVEL COMPONENTS OF MITOCHONDRIAL REGULATION OF INSULIN SECRETION IN TYPE 2 DIABETES - PROJECT SUMMARY THE SUCCESS OF PREVENTIVE AND SUPPORTIVE APPROACHES IN TYPE 2 DIABETES MELLITUS (T2D) DEPENDS ON THE SPONTANEOUS RECOVERY OF Β-CELL FUNCTION. A CENTRAL COMPONENT OF Β-CELL FUNCTION, GLUCOSE METABOLISM, IS IMPAIRED IN T2D. ITS CAUSES, AND HOW THIS IMPAIRMENT MIGHT LEAD TO FUNCTIONAL FAILURE OF Β-CELLS, REMAIN UNKNOWN. UNTIL DELINEATED, TARGETING OF THE DYSFUNCTIONAL PATHWAYS AND ADDRESSING THE CAUSE OF THE AB- NORMALITY ARE UNLIKELY AND THERAPIES REMAIN SUPPORTIVE RATHER THAN DISEASE-MODIFYING. THE OVERALL OBJEC- TIVE IS TO IDENTIFY COMPONENTS OF HUMAN Β-CELL METABOLISM THAT ARE CRITICAL FOR INSULIN SECRETION IN HEALTHY INDIVIDUALS AND FAIL TO SUPPORT PHYSIOLOGICAL SECRETION IN T2D, AND TO IDENTIFY COMPONENTS MEDIATING Β-CELL COMPENSATION. WE HAVE IDENTIFIED A NOVEL INTRAMITOCHONDRIAL METABOLIC ACTIVATION PHENOMENON AS SUCH A COMPONENT, AND INTEND TO DETERMINE ITS MECHANISM AND ROLE. THE CENTRAL HYPOTHESIS OF THE PROJECT IS THAT IN T2D Β-CELLS A METABOLIC REARRANGEMENT IMPAIRS THE RESPONSE OF Β-CELLS TO GLUCOSE BY LIMITING THE ACTI- VATION OF SUCCINATE DEHYDROGENASE (SDH), AND TRIGGERS AN UPREGULATION OF AMPLIFICATION PATHWAYS, ALTER- ING THE PHYSIOLOGICAL DYNAMICS OF INSULIN SECRETION IN T2D. USING HUMAN ORGAN DONOR ISLETS AND MICRO- SCALE BIOENERGETIC AND CELL PHYSIOLOGY ASSAYING TOGETHER WITH GENETIC (CRISPR) AND PHARMACOLOGICAL MODULATION OF METABOLISM, 1) THE MECHANISM OF BIOENERGETIC ACTIVATION OF NORMAL HUMAN Β-CELLS DURING GLUCOSE-STIMULATED INSULIN SECRETION (GSIS) WILL BE IDENTIFIED. WE HYPOTHESIZE THAT THE ACTIVATION OF SDH IS REQUIRED FOR GSIS, AND THIS IS CONVEYED BY METABOLIC COUPLING FACTOR PATHWAYS THROUGH REGULATING MA- TRIX OXALOACETATE, MALONATE OR THE FLUX CONTROL BY SDH. 2) THE MECHANISM OF T2D Β-CELL COMPENSATION AND ITS EFFECTS IN THE CONTEXT OF IMPAIRED BIOENERGETIC ACTIVATION WILL BE DETERMINED. HERE, OUR HYPOTHESIS IS THAT T2D Β-CELLS COMPENSATE FOR THE LOSS OF BIOENERGETIC CONTROL BY UPREGULATING AMPLIFICATION PATH- WAYS, AND THIS ALTERS CHARACTERISTICS OF SECRETION. 3) THE BETA-CELL-SPECIFIC GENE EXPRESSION PATTERN RE- QUIRED FOR THE BIOENERGETIC ACTIVATION AND COMPENSATION WILL BE DETERMINED. OUR HYPOTHESIS IS THAT THE MITOCHONDRIAL ACTIVATION MECHANISM IS A NETWORK-LEVEL PHENOMENON, REQUIRING A PARTICULAR GENE EXPRES- SION PATTERN. HARNESSING CELL-TO-CELL AND INDIVIDUAL-TO-INDIVIDUAL HETEROGENEITY, MITOCHONDRIAL FUNCTION AND NUCLEAR GENE EXPRESSION WILL BE CORRELATED ON THE SINGLE-CELL LEVEL. THE PROPOSED RESEARCH REPRESENTS A SUBSTANTIAL DEPARTURE FROM THE STATUS QUO BY RECOGNIZING A NOVEL MITOCHONDRIAL REGULATION MECHANISM AS THE CONTROLLING ENTITY OF GLUCOSE METABOLISM AND INSULIN SECRETION. FURTHERMORE, IT DEPARTS FROM THE TRADI- TIONAL DICHOTOMY OF TRIGGERING AND AMPLIFICATION PATHWAYS CONTROLLING GSIS BY DETERMINING HOW THESE PATHWAYS CONVERGE IN BIOENERGETIC REGULATION. THIS WORK WILL BE SIGNIFICANT BECAUSE COMBINING THE IDENTI- FICATION OF ABNORMALITIES IN T2D Β-CELL ENERGY METABOLISM WITH KNOWLEDGE OF RELATED COMPENSATORY MECH- ANISMS, WILL ALLOW TARGETING OF THESE PATHWAYS TO SUPPORT Β-CELL FUNCTION.

NUCLEAR LAMIN FUNCTIONS AND HUMAN DISEASE

NOVEL MITOCHONDRIA-TO-LYSOSOME CROSSTALK CONTRIBUTES TO LYSOSOMAL DYSFUNCTION DURING AGING - PROJECT SUMMARY/ABSTRACT AGING IS ACCOMPANIED BY THE GRADUAL DETERIORATION WITHIN EACH INDIVIDUAL CELLULAR COMPARTMENTS/ORGANELLES AS WELL AS THE COLLAPSE OF THE INTERCONNECTIONS AMONG THEM, LEADING TO THE ESTABLISHMENT OF DIFFERENT HALLMARKS OF AGING. HOWEVER, THESE HALLMARKS OF AGING, INCLUDING LOSS OF PROTEOSTASIS, MITOCHONDRIAL DYSFUNCTION, AND CELLULAR SENESCENCE, ARE TRADITIONALLY STUDIED SEPARATELY AND LEAVE THE CONNECTIONS AMONG THESE HALLMARKS LARGELY UN- KNOWN. WITH A GROWING UNDERSTANDING OF THESE INTER-ORGANELLE INTERACTIONS WITHIN YOUNG CELLS, IT HAS BECOME OF INTEREST TO EXPLORE HOW LOSSES IN COMPARTMENTAL CROSSTALK CONTRIBUTE TO THE AGE-ASSOCIATED DEGENERATION OF OR- GANELLE FUNCTION AND THE ESTABLISHMENT/CONNECTION OF HALLMARKS OF AGING. WE RECENTLY DISCOVERED AN UNEXPECTED CROSSTALK BETWEEN MITOCHONDRIA AND THE LYSOSOME (VACUOLE IN YEAST) THAT EXPLAINS AGE-ASSOCIATED VACUOLE/LYSOSOME DE-ACIDIFICATION, A PROCESS CONSERVED FROM YEAST TO HUMAN CELLS AND WHICH CONTRIBUTES TO THE LOSS OF PROTEOSTASIS DURING AGING AND IN MANY AGE-RELATED DISEASES. WE FOUND THAT MITOCHONDRIA CONTRIBUTE TO THE ACIDIFICATION OF LYSOSOME/VACUOLE VIA MEMBRANE CONTACT SITES. MOREOVER, OUR PRE- LIMINARY STUDIES SUGGEST THAT THE COLLAPSE OF MITOCHONDRIA-VACUOLE CONTACT DURING THE REPLICATIVE AGING OF YEAST CAUSES AGE-DEPENDENT DE-ACIDIFICATION OF VACUOLE. SIMILARLY, WE OBSERVED A CLOSE CORRELATION BETWEEN MITOCHON- DRIA AND LYSOSOME ACIDIFICATION IN CULTURED HUMAN CELLS AND IN C. ELEGANS, SUGGESTING EVOLUTIONARY CONSERVATION OF THIS PROCESS. TO ADDRESS HOW AGING AFFECTS THE MITOCHONDRIA-LYSOSOME/VACUOLE CONTACT AND WHETHER THIS MI- TOCHONDRIA-TO-LYSOSOME AXIS PLAYS A CRITICAL ROLE IN LYSOSOME ACIDIFICATION IN CELLULAR SENESCENCE AND ANIMALS, WE HERE PROPOSE TO INVESTIGATE THE MITOCHONDRIA-LYSOSOME/VACUOLE CONTACT AND LYSOSOME/VACUOLE ACIDIFICATION DUR- ING AGING AND IN DIFFERENT LONGEVITY PARADIGMS. SPECIFICALLY, WE WILL COMBINE THE EXPERTISE OF OUR DIFFERENT MODEL SYSTEMS TO (1) DISSECT THE MECHANISM(S) UNDERLYING THE AGE-RELATED LOSS OF MITOCHONDRIA-VACUOLE CONNECTION IN YEAST; (2) DETERMINE MITOCHONDRIAL CONTRIBUTION TO LYSOSOME ACIDIFICATION IN HUMAN CELLS; AND (3) DISSECT THE ROLES OF MITOCHONDRIA-LYSOSOME CONNECTION IN LYSOSOME ACIDIFICATION IN C. ELEGANS. OUR STUDIES WILL INVESTIGATE A NOVEL, POSSIBLY EVOLUTIONARILY CONSERVED INTER-ORGANELLE COMMUNICATION CONNECT- ING MITOCHONDRIAL DYSFUNCTION AND LYSOSOME/VACUOLE DE-ACIDIFICATION ACROSS DIFFERENT ORGANISMS AND LONGEVITY PARADIGMS. ALTHOUGH TRADITIONAL THINKING HAS HELD THAT AGE-ASSOCIATED LYSOSOME/VACUOLE DYSFUNCTION IS RESPON- SIBLE FOR INDUCING MITOCHONDRIAL DYSFUNCTION IN A UNI-DIRECTIONAL MANNER, OUR STUDY WILL DETERMINE WHETHER MITO- CHONDRIAL DYSFUNCTION MAY CONVERSELY IMPACT ON LYSOSOME/VACUOLE DYSFUNCTION DURING AGING REVEALING A HERE- TO-FORE UNAPPRECIATED TWO-WAY CROSSTALK BETWEEN THESE TIGHTLY CONNECTED ORGANELLES WHICH MAY TOGETHER CON- TRIBUTE TO SEVERAL IMPORTANT HALLMARKS OF AGING. AS SUCH, THE LONG-TERM IMPLICATIONS OF THIS STUDY COULD BE THE IDENTIFICATION OF NOVEL CONSERVED THERAPEUTIC TARGETS FOR AGE-RELATED DISEASES WITH LYSOSOME DEFECTS.

EVALUATING DIVERSE TECHNOLOGIES FOR DETECTING AND VALIDATING SENESCENT CELLS IN VIVO - PROJECT SUMMARY SINGLE CELL PROFILING HAS RECENTLY EXPLODED INTO MAINSTREAM BIOLOGY. SINGLE CELL PROFILING HAS BEEN EMPLOYED IN MYRIAD APPLICATIONS IN BIOLOGY, INCLUDING MULTIPLE DISEASES, EMBRYONIC DEVELOPMENT, COMPARATIVE EVOLUTIONARY STUDIES AND AGING. MOST RECENTLY, SINGLE CELL PROFILING HAS INFORMED THE PHENOMENA OF CELLULAR SENESCENCE. THIS PROPOSAL HAS TWO PHASES, UG3 (MODEL SYSTEMS), AND UH3 (HUMAN VALIDATION). CELLULAR SENESCENCE IS A MULTI-FACETED CELL FATE THAT ARRESTS CELL PROLIFERATION, ESSENTIALLY IRREVERSIBLY, AND ACTIVATES THE PRODUCTION AND SECRETION OF PRO-INFLAMMATORY CYTOKINES, CHEMOKINES, GROWTH FACTORS, PROTEASES AND LIPIDS, TERMED THE SENESCENCE ASSOCIATED SECRETORY PHENOTYPE (SASP). THE SASP CAN INFLUENCE TISSUE MICROENVIRONMENTS, AND THUS SENESCENT CELLS CAN STRONGLY AFFECT TISSUE FUNCTION AND LIKELY THE SYSTEMIC MILIEU. SENESCENT CELLS INCREASE WITH AGE AND CAN DRIVE A GROWING LIST OF AGE-RELATED PATHOLOGIES, RANGING FROM NEURODEGENERATION TO CANCER, IN PART THROUGH THE SASP. THERE IS INCREASING EVIDENCE THAT THERE ARE NO UNIVERSAL MARKERS FOR SENESCENT CELLS. INSTEAD, SENESCENT CELLS, WHILE SHARING CERTAIN CHARACTERISTICS AND BIOMARKERS, ARE REMARKABLY HETEROGENEOUS, VARYING IN CHARACTERISTICS WITH GENOTYPE, CELL AND TISSUE TYPE, SENESCENCE INDUCER, TISSUE (AND CELL CULTURE) MICROENVIRONMENT, AND CHRONOLOGY (TIME AFTER INITIAL SENESCENCE INDUCTION). WHILE SOME OF THE MORE COMMONLY EMPLOYED SENESCENCE MARKERS HAVE UTILITY IN SUPERFICIALLY IDENTIFYING SENESCENT CELLS DE NOVO, THE ONUS REMAINS ON THE INVESTIGATOR TO DEMONSTRATE WHY A CELL SHOULD BE CONSIDERED SENESCENT, RATHER THAN RELYING ON HISTORICAL MARKERS SUCH AS P16INK4A OR P21CIP1. THUS, NEW TECHNOLOGIES DESIGNED TO IDENTIFY NOVEL SENESCENT CELLS AND PHENOTYPES ARE NECESSARY THAT WILL REQUIRE VALIDATION BOTH IN CULTURE AND IN TISSUE. THE ULTIMATE GOAL OF THIS PROPOSAL IS TO DEVELOP NEW TECHNOLOGIES TO MAP SENESCENT SIGNATURES BACK TO INTACT HUMAN TISSUE. THIS GOAL WILL ENABLE US TO IDENTIFY AND SPATIALLY CHARACTERIZE SENESCENT CELLS IN EACH TISSUE, UNCOVERING UNIQUE MARKERS DEPENDING ON TISSUE AND CELL TYPE. PILOT 1: IDENTIFY SENESCENT CELLS IN THE MOUSE VASCULATURE, AND DETERMINE IF THEY CAN BE DETECTED BY ULTRASOUND, AND VERIFIED IN HUMAN TISSUE; PILOT 2: DEVELOP A MICROPHYSIOLOGIC EX VIVO TISSUE-ON-A-CHIP TO MODEL OVARIAN SENESCENCE, AND HUMAN TISSUE-TISSUE INTERACTIONS VIA THE SASP. THESE TWO PILOTS WILL USE A COMBINATION OF CELL SURFACE MARKERS IDENTIFIED IN OUR INITIAL PROFILING, AND DIGITAL SPATIAL PROFILING (DSP, NANOSTRING) OR VISIUM (10X) TO LOCALIZE SENESCENT SIGNATURES TO MORPHOLOGICAL STRUCTURES AND CELLS IN TISSUE SECTIONS. OUR PROPOSAL WILL DEVELOP VALIDATED MARKERS OF SENESCENCE IN MULTIPLE TISSUES AND CELL TYPES NOT PREVIOUSLY CHARACTERIZED TO DEPLOY THESE TECHNOLOGIES TO THE BROADER COMMUNITY.

CELLULAR SENESCENCE AND ALZHEIMER'S DISEASE

PROTEIN INTERACTIONS AND PROTEIN CONFORMATION IN AGING AND DISEASE (6 OF 11)

HDACS IN NEURODEGENERATION AND AGING

DEFECTIVE ESTROGEN RECEPTORS IN BREAST TUMORS

TIMSTOF FLEX MALDI-2 FOR SPATIAL MOLECULAR IMAGING - PROJECT SUMMARY/ABSTRACT THIS SHARED INSTRUMENTATION PROPOSAL SEEKS FUNDING FOR THE PURCHASE OF A BRUKER ‘TIMSTOF FLEX MALDI-2 FOR SPATIAL MOLECULAR IMAGING’ INCLUDING A BUNDLED HTX M3+ SPRAYER. THIS NEW MASS SPECTROMETRY (MS) PLATFORM WILL FACILITATE ONGOING AND FUTURE SCIENTIFIC RESEARCH TO SPATIALLY RESOLVE MOLECULAR SIGNATURES, EVEN IN A MULTI-OMIC WORKFLOW MONITORING PROTEINS, GLYCANS, LIPIDS, AND METABOLITES ALL WITHIN THE SAME TISSUE DOWN TO A SPATIAL RESOLUTION OF ~5-10 ΜM. ALL COSTS WILL BE ACCOUNTED FOR WITH THIS PROPOSAL. WE PROPOSE TO OPERATE THIS INTEGRATED TIMSTOF FLEX MALDI-2 MASS SPECTROMETRY SYSTEM EXCLUSIVELY FOR SPATIALLY RESOLVED MOLECULAR IMAGING OF HUMAN AND ANIMAL TISSUES AND ORGANOIDS AS PART OF LARGE GROUP OF 18 INDEPENDENT PRINCIPAL INVESTIGATORS FROM THE BUCK INSTITUTE AND SEVEN OTHER INSTITUTIONS OR UNIVERSITIES. THESE RESEARCHERS BRING TOGETHER 19 NIH-FUNDED GRANTS (INCLUDING P01, U54, U01, R01, P30, UG3 ACTIVITY CODES) AND ONE R01 SUPPLEMENT, ALL OF WHICH HAVE SIGNIFICANT NEEDS FOR STATE-OF-THE-ART MASS SPECTROMETRY FOR SPATIALLY RESOLVED PROTEOMICS, METABOLOMICS, LIPIDOMICS AND GLYCAN ANALYSIS – EVEN IN A MULTI-OMIC SYSTEMS APPROACH. THE MOST CRITICAL FEATURES OF THIS NEW INSTRUMENTATION WILL BE THE 1) SECOND POST-IONIZATION LASER (MALDI-2) TO GREATLY REDUCE NOISE AND TO IMPROVE IONIZATION AND SENSITIVITY SUBSTANTIALLY, 2) FULL MS IMAGING WORKFLOW SOLUTIONS WITH WORKFLOWS USING THE HTX M3+ SPRAYER BUNDLE FOR SAMPLE PREPARATION AND MATRIX APPLICATION REQUIRED FOR MS IMAGING, 3) HIGH SPATIAL RESOLUTION TO REVEAL SMALL DISTINCT ARCHITECTURAL FEATURES IN TISSUES, 4) DUAL TIMS ION MOBILITY FOR REDUCTION OF COMPLEXITY, SEPARATION OF ISOBARIC FEATURES AND INCREASED SENSITIVITY AND SELECTIVITY FOR IMPROVED MS IMAGING, 5) LIPID AND METABOLITE MS IMAGING USING ION MOBILITY CROSS SECTION (CCS) MEASUREMENTS AS ADDITIONAL IDENTIFIER (CCS) USING THE METABOSCAPE DATABASES AND SCILS (COMBINATION OF M/Z AND CCS IN SPECTRAL LIBRARIES AND DATABASES), AND 6) TRUE MULTI-OMIC APPROACH FOR MS IMAGING. OVERALL, THE ACQUISITION OF THIS INSTRUMENT TO THE MASS SPECTROMETRY CORE AT THE BUCK INSTITUTE FOR RESEARCH ON AGING WILL PROVIDE UNIQUE AND NOVEL FEATURES THAT WILL BRING INNOVATION AND TOTALLY NOVEL CAPABILITIES (SPATIAL MOLECULAR MS IMAGING) FOR OUR 18 PROPOSED USER PROJECTS THAT ARE EXAMINING KEY ISSUES OF HUMAN HEALTH AND HEALTH SPAN, INCLUDING THE THIS INNOVATIVE MASS SPECTROMETRY MOLECULAR IMAGING TECHNOLOGY TO STUDY THE BIOLOGY OF AGING INCLUDING DIFFERENTIAL AGING OF SPECIFIC ORGANS AND TO INVESTIGATE AGE-RELATED DISEASES, INCLUDING ALZHEIMER’S AND HUNTINGTON’S DISEASE, A PARKINSONISM, OSTEOARTHRITIS, KIDNEY DISEASES AND METABOLIC DISORDERS.

TOR, TRANSLATION AND AGING

INVESTIGATION OF IMPAIRED NEURAL STEM CELL ACTIVATION IN ALZHEIMER'S DISEASE - PROJECT SUMMARY ALZHEIMER’S DISEASE (AD) IS A DEVASTATING NEURODEGENERATIVE DISEASE THAT CAUSES SEVERE ATROPHY IN THE HIPPOCAMPAL AREA. THE DENTATE GYRUS OF THE MAMMALIAN HIPPOCAMPUS HARBORS POPULATIONS OF QUIESCENT NEURAL STEM CELLS (NSCS) THAT CAN RE-ENTER THE CELL CYCLE AND DIFFERENTIATE INTO FUNCTIONAL EXCITATORY NEURONS. THIS PROCESS OF NEUROGENESIS SUPPORTS LEARNING AND MEMORY FUNCTIONS, AS WELL AS HEALTHY MOOD REGULATION. HOWEVER, IN AGING AND AD, LEVELS OF NEUROGENESIS SHARPLY DECLINE, AND WORK IN RODENTS HAS DEMONSTRATED A FUNCTIONAL LINK BETWEEN LEVELS OF NEUROGENESIS AND COGNITIVE PERFORMANCE. ALTHOUGH QUIESCENT NSCS ARE THE SOURCE OF NEW NEURONS, THE CURRENT UNDERSTANDING OF HOW THESE CELLS ARE IMPACTED IN AD IS EXTREMELY LIMITED. PRELIMINARY WORK SHOWS THAT QUIESCENT NSCS ARE DEFECTIVE IN A MOUSE MODEL OF AD. MOREOVER, THESE CELLS ARE PARTICULARLY VULNERABLE TO DAMAGE DURING AGING, WHICH IS THE GREATEST RISK FACTOR FOR AD. THIS PROPOSAL TAKES AN INTEGRATED, MULTIOMICS APPROACH TO ADDRESS THE CRITICAL QUESTION OF HOW THE QUIESCENT POOL IS AFFECTED IN AD. USING A WELL-ESTABLISHED MOUSE MODEL AND HUMAN TISSUE, SPECIFIC AIM 1 WILL DEFINE THE TRANSCRIPTOMIC AND EPIGENOMIC CHANGES AT THE SINGLE CELL LEVEL IN AD. WORK IN AIM 2 WILL EMPLOY METABOLOMICS TO FULLY ELUCIDATE THE METABOLIC CHANGES THAT OCCUR IN AD. LASTLY, AIM 3 WILL USE A FUNCTIONAL TRANSCRIPTOMICS SCREEN TARGETING CENTRAL PATHWAYS IN QUIESCENT NSCS TO IDENTIFY HOW THESE CELLS ARE SELECTIVELY VULNERABLE IN AD. TOGETHER THIS WORK WILL RESULT IN A COMPREHENSIVE UNDERSTANDING OF HOW DORMANT NSCS ARE AFFECTED IN A MOUSE MODEL OF AD AS WELL AS IN HUMAN TISSUE. THIS STUDY WILL PROVIDE KEY DATA TO THE NEURODEGENERATION COMMUNITY THAT CAN BE LEVERAGED FOR FUNCTIONAL STUDIES AND THE DEVELOPMENT OF INTERVENTIONS TO PREVENT, TREAT AND EVEN CURE NEURODEGENERATION IN THE LONG TERM.

SUBCELLULAR LOCALIZATION OF ALPHA-SYNUCLEIN AND ITS IMPACT ON NEURODEGENERATION

NEURONAL STEM CELLS AND AGING

MATRIX METALLOPROTEINASES: THERAPEUTIC TARGETS FOR HUNTINGTON'S DISEASE

HUNTINGTIN INTERACTING PROTEINS AS MODIFIERS OF HUNTINGTON'S DISEASE

BASIC MECHANISMS IN AGING AND AGE RELATED DISEASE

CELLULAR AND CIRCUIT MECHANISMS OF NEUROPEPTIDE SIGNALING - PROJECT SUMMARY THE OVERALL GOAL OF NIGMS-FUNDED RESEARCH IN MY LAB IS TO UNDERSTAND THE CELLULAR AND CIRCUIT MECHANISMS BY WHICH NEUROPEPTIDE SIGNALING INFLUENCES THE NEURAL CIRCUITS THAT CONTROL BEHAVIOR. NEUROPEPTIDES ARE THE MOST COMMON SIGNALING MOLECULE IN THE CENTRAL NERVOUS SYSTEM AND AMONG THE BEST MARKERS FOR CELL TYPES IN THE BRAIN. THESE ENDOGENOUS PEPTIDES TRANSMIT MESSAGES WITHIN THE BRAIN AND ACROSS THE BODY TO CONTROL VITAL PHYSIOLOGIC PROCESSES LIKE ENERGY HOMEOSTASIS, AS WELL AS MOTIVATIONAL AND EMOTIONAL STATES INCLUDING SLEEP, AROUSAL, PAIN, STRESS, AND MOOD. DYSREGULATION OF PEPTIDERGIC SIGNALING IS IMPLICATED IN MEDICAL CONDITIONS RANGING FROM OBESITY TO PSYCHIATRIC DISORDERS. A VAST SCIENTIFIC LITERATURE HAS INVESTIGATED THE ROLE OF NEUROPEPTIDES IN PHYSIOLOGY AND BEHAVIOR OVER DECADES, YET EVEN FOR MANY WELL-STUDIED CIRCUITS, THEIR FUNCTIONAL SIGNIFICANCE IS STILL AN OPEN QUESTION. BECAUSE THEY ARE NOT RESTRICTED SPATIALLY BY THE ANATOMICAL WIRING DIAGRAM NOR TEMPORALLY BY RAPID RE-UPTAKE OR DEGRADATION, DELINEATING THE PRECISE CONNECTIVITY OF NEUROPEPTIDE CIRCUITS IN WHOLE ANIMALS HAS PROVED CHALLENGING. MY LAB SEEKS TO ADDRESS THIS GAP BY DEVELOPING NOVEL METHODS TO MONITOR AND SELECTIVELY MANIPULATE NEUROPEPTIDE SIGNALING IN LIVING ANIMALS, AND TO IDENTIFY THE FUNDAMENTAL ENZYMES THAT REGULATE NEUROPEPTIDE COMMUNICATION. WE EMPLOY ADVANCED IMAGING, GENETIC, AND BIOCHEMICAL APPROACHES TO INVESTIGATE THESE QUESTIONS USING BOTH WORM AND MOUSE MODEL SYSTEMS FROM THE SUBCELLULAR LEVEL ALL THE WAY TO BEHAVIORAL OUTPUT. OUR GOAL IS TO UNDERSTAND THE BIOCHEMICAL LOGIC OF CIRCUIT NEUROMODULATION BY NEUROPEPTIDES WITH THE SAME CLARITY THAT WE UNDERSTAND FAST NEUROTRANSMISSION AT SYNAPSES. THIS RESEARCH WILL PROVIDE FUNDAMENTAL NEW INSIGHT INTO LONGSTANDING QUESTIONS ABOUT THE SPATIAL AND TEMPORAL ORGANIZATION OF NEUROPEPTIDE SIGNALING AND LEAD TO AN UNDERSTANDING OF HOW LONG-TERM CHANGES ARE AFFECTED IN THE NERVOUS SYSTEM THAT RESULT IN DIFFERENT BEHAVIORS. AS NEUROPEPTIDES AND THEIR RECEPTORS ARE DRUGGABLE TARGETS IMPLICATED IN A WIDE-RANGE OF DISEASES, WE ANTICIPATE THAT MECHANISTIC INSIGHT INTO THEIR SIGNALING IS LIKELY TO HAVE BROAD CLINICAL RELEVANCE FOR DISEASES CHARACTERIZED BY BEHAVIORAL DYSFUNCTION.

CHECKPOINT FUNCTIONS, LIFESPAN DETERMINATION AND NEURODEGENERATION (3 OF 11)

CA2+ SIGNALING AND STEM CELL DYNAMICS

NOVEL PRIONIC MECHANISM UNDERLYING ALZHEIMER?S DISEASE

ROLE OF MRNA TRANSLATION IN THE EFFECTS OF DIETARY RESTRICTION ON LIFESPAN

TAKEOFF: TARGETING AGING WITH KETONE ESTER IN OLDER ADULTS FOR FUNCTION IN FRAILTY - PROJECT SUMMARY/ABSTRACT THE GEROSCIENCE APPROACH OF MODULATING FUNDAMENTAL AGING MECHANISMS HOLDS GREAT PROMISE FOR GENERATING EFFECTIVE NEW THERAPEUTICS FOR COMPLEX, MULTIFACTORIAL CONDITIONS OF AGING SUCH AS FRAILTY WHICH CONTRIBUTE SO MUCH TO FUNCTIONAL DECLINE, DISABILITY, AND LOSS OF INDEPENDENCE IN OLDER ADULTS. THE FRAILTY SYNDROME IS CONCEPTUALIZED AS A CONDITION OF PROGRESSIVE FUNCTIONAL DECLINE AND INCREASED VULNERABILITY TO STRESSORS RESULTING FROM DECREASED FUNCTIONAL RESERVE. THE PATHOPHYSIOLOGY OF FRAILTY IS COMPLEX AND NOT WELL UNDERSTOOD, BUT IS GENERALLY THOUGHT TO PROMINENTLY INCLUDE CELLULAR ENERGY PRODUCTION DEFICITS ALONG WITH CHRONIC INFLAMMATION AND IMMUNE DYSFUNCTION. DIETARY RESTRICTION AND FASTING, INTERVENTIONS CLOSELY LINKED TO FUNDAMENTAL MECHANISMS OF AGING, PROMOTE THE ENDOGENOUS PRODUCTION OF KETONE BODIES AS A MEANS OF SUPPLYING FAT-DERIVED ENERGY TO TISSUES. NEW ADVANCES IN UNDERSTANDING THE BIOLOGICAL ACTIVITIES OF THE KETONE BODY BETA-HYDROXYBUTYRATE (BHB) SUGGEST THAT BOTH ENERGY PRODUCTION AND SIGNALING ACTIVITIES OF BHB MAY HAVE A MECHANISTIC ROLE IN MODULATING AGING, AND MAY BE PARTICULARLY RELEVANT TO THE PATHOPHYSIOLOGY OF FRAILTY. KETONE ESTERS HAVE RECENTLY EMERGED AS A PHARMACOLOGICAL MEANS OF DELIVERING KETONE BODIES. WE HAVE ASSEMBLED A MULTIDISCIPLINARY TEAM OF EXPERTS IN GEROSCIENCE, GERIATRICS AND FRAILTY, KETONE BODY BIOLOGY, IMMUNOSENESCENCE, AND KETONE BODY-RELATED CLINICAL TRIALS TO CARRY OUT A MULTI-SITE, PROOF-OF-CONCEPT PHASE 2A CLINICAL TRIAL OF A KETONE ESTER (KE) TARGETING FRAILTY IN PREFRAIL AND MILDLY FRAIL OLDER ADULTS. TAKEOFF IS A PROSPECTIVE, RANDOMIZED, DOUBLE-BLIND, 12-WEEK STUDY OF KETONE ESTER VS. MATCHED PLACEBO. AIM 1 WILL TEST IF MUSCLE STRENGTH AND OTHER CLINICAL FRAILTY- AND AGING-RELATED OUTCOMES IMPROVE WITH KE. AIM 2 WILL DETERMINE THE SAFETY AND TOLERABILITY OF DAILY KE IN BROADLY REPRESENTATIVE PRE-FRAIL AND FRAIL OLDER ADULTS. AIM 3 WILL TEST THE HYPOTHESIS THAT KETONE ESTER IMPACTS THE AGING IMMUNE SYSTEM, SKELETAL MUSCLE, AND BIOMARKERS OF AGING VIA DETAILED MECHANISTIC IMMUNOPHENOTYPING, MUSCLE METABOLISM, AND BIOMARKER STUDIES THAT LEVERAGE THE STRONG SPECIALIST EXPERTISE AT THE PARTICIPATING INSTITUTIONS. ALTOGETHER, THIS TRANSLATIONAL EFFORT WILL PROVIDE CRUCIAL INFORMATION ON THE SAFETY AND EFFICACY OF INTERVENTIONS INCREASINGLY BEING USED BY THE PUBLIC DESPITE A PAUCITY OF DATA, AND WITH POTENTIAL TO IMPACT IMPORTANT AGING-RELATED CONDITIONS TO IMPROVE THE HEALTH AND INDEPENDENCE OF OLDER ADULTS.

SUPPORT AND PREPARATION FOR AN ALZHEIMER'S RESEARCH CAREER - PROJECT SUMMARY/ABSTRACT ALZHEIMER'S DISEASE AND RELATED DEMENTIAS (ADRD) ARE THE LEADING CAUSE OF DEMENTIA IN THOSE OVER THE AGE OF 65, LEADING TO PROGRESSIVE MEMORY LOSS AND COGNITIVE DECLINE. ALTHOUGH THE NATIONAL PLAN TO ADDRESS AD SEEKS TO ACCELERATE RESEARCH TOWARD TREATMENTS, THERE IS A SHORTAGE OF SCIENTISTS NECESSARY TO CONDUCT A WIDE VARIETY OF INTERDISCIPLINARY RESEARCH PROJECTS. THE BUCK INSTITUTE'S SUPPORT AND PREPARATION FOR AN ALZHEIMER'S RESEARCH CAREER (SPARC) PROGRAM WILL ADDRESS THIS NEED BY RECRUITING A DIVERSE GROUP OF POSTBACCALAUREATES AND TRAINING THEM IN A GEROSCIENCE-INSPIRED APPROACH TO ADRD RESEARCH. THIS INTERDISCIPLINARY APPROACH WILL INVESTIGATE THE CONTRIBUTION OF COMMON PROCESSES GOVERNING AGING ON THE DEVELOPMENT OF ADRD. THE DIVERSE RESEARCH EXPERTISE AND COLLABORATIVE CULTURE OF THE BUCK INSTITUTE PROVIDE A UNIQUE TRAINING ENVIRONMENT TO ENSURE THE NEXT GENERATION OF RESEARCHERS IN THE FIELD ARE PROVIDED THE KNOWLEDGE AND SKILLS TO REDUCE THE BURDEN OF ADRD. THE SPARC PROGRAM WILL BE DESIGNED TO INCREASE PARTICIPATION OF STUDENTS FROM UNDERREPRESENTED GROUPS IN ADRD RESEARCH AND PREPARE THEM FOR ADDITIONAL TRAINING IN ADVANCED DEGREE PROGRAMS OR PRIVATE SECTOR RESEARCH CAREERS IN AD-RELATED DISCIPLINES. RECENT COLLEGE GRADUATES WILL BE RECRUITED USING COLLEGE AND UNIVERSITY ONLINE PROGRAMS, SUCH AS HANDSHAKE AND SIMPLICITY. PARTNERSHIPS WITH NON-PROFIT ORGANIZATIONS THAT SUPPORT UNDERREPRESENTED STUDENTS, SUCH AS STUDENTS RISING ABOVE AND BIOTECH PARTNERS, WILL ENHANCE RECRUITING EFFORTS. A ONE-MONTH INTRODUCTORY TRAINING PERIOD WILL FAMILIARIZE POSTBACCALAUREATES TO CORE RESEARCH CONCEPTS, AND SHADOWING OPPORTUNITIES IN PROSPECTIVE HOST LABS WILL ENSURE STRONG MENTOR-MENTEE MATCHES DURING THE ELEVEN-MONTH MENTORED RESEARCH PROJECT, WHICH WILL CONTINUE TO BE BOLSTERED BY TRAINING ACTIVITIES. FOREMOST, A MOCK GRANT PROPOSAL WRITING ASSIGNMENT AND MOCK STUDY SECTION WILL PROVIDE FRAMEWORK FOR SPARC RESEARCHERS TO DIGEST LITERATURE ASSOCIATED WITH THEIR PROJECT AS WELL AS DEVELOP THEIR SCIENTIFIC WRITING SKILLS. ORAL PRESENTATION SKILLS WILL BE PRACTICED WITH PRESENTATIONS TO BOTH A LAY AND SCIENTIFIC AUDIENCE. IN ADDITION TO SERVING AS A TRAINING OPPORTUNITY FOR SPARC RESEARCHERS, THE LAY AUDIENCE PRESENTATION ALSO WILL PROVIDE AN OUTREACH OPPORTUNITY TO INTRODUCE LOCAL HIGH SCHOOL AND COLLEGE STUDENTS TO ADRD RESEARCH AND CAREER OPPORTUNITIES. THE SCIENTIFIC WRITING AND PRESENTATION ASSIGNMENTS NOT ONLY WILL HELP SPARC RESEARCHERS UNDERSTAND THEIR PROJECTS BETTER BUT ALSO PREPARE THEM FOR DISCUSSION OF THEIR RESEARCH IN FUTURE APPLICATIONS AND INTERVIEWS. MENTORS WILL PROVIDE ADDITIONAL CAREER SUPPORT BOTH DURING AND AFTER COMPLETION OF THE PROGRAM THROUGH APPLICATION FEEDBACK, INTERVIEW PRACTICE, AND NETWORKING INTRODUCTIONS. TRAINING ALUMNI CAN PROVIDE ADDITIONAL PEER SUPPORT THROUGH JOB OR ADVANCED DEGREE PROGRAM ADVICE, APPLICATION OR INTERVIEW SUGGESTIONS, AND ALTERNATIVE CAREER OPTIONS. IN SUMMARY, THE TRAINING PROVIDED BY THE BUCK INSTITUTE'S SPARC PROGRAM WILL RECRUIT AND PREPARE A COHORT OF STUDENTS FROM UNDERREPRESENTED GROUPS FOR FURTHER ADVANCED STUDIES OR A CAREER IN GEROSCIENCE-INSPIRED ADRD RESEARCH.

INTESTINAL HOMEOSTASIS AND AGING

DEVELOPMENT OF THE OLEAGINOUS YEAST RHODOSPORIDIUM TORULOIDES AS A NEW MODEL ORGANISM FOR A SYSTEMS-LEVEL ANALYSIS OF LIPID PRODUCTIVITY

ORBITRAP FUSION LUMOS ETD WITH FAIMS PRO

S6 KINASE, AGING AND AGE-RELATED DISEASE

EPIGENETIC REGULATION OF HEALTHSPAN AND LONGEVITY BY KETONE BODIES

FUNCTIONAL CONTRIBUTION OF NEURAL STEM CELLS IN STROKE

TOR, MRNA TRANSLATION AND DIETARY RESTRICTION IN DROSOPHILA

IMPROVING ER PROTEOSTASIS AS A THERAPEUTIC STRATEGY TO TREAT ALS

MITOCHONDRIAL SITES OF REACTIVE OXYGEN SPECIES GENERATION IN CELLS.

PERIPHERAL PROTEOSTASIS AND PROGRESSION OF AD RELATED NEURODEGENERATION: A GUT-MUSCLE-BRAIN TRIANGLE

DETERMINING THE ROLE OF OXR1 IN AGING AND ALZHEIMER'S DISEASE - PROJECT SUMMARY/ABSTRACT DESPITE IDENTIFYING SOME OF THE GENETIC RISK FACTORS FOR AD, THE PRECISE ETIOLOGY OF MOST CASES OF LATE-ONSET ALZHEIMER'S DISEASE (LOAD) IS UNKNOWN. SUBSEQUENTLY, THERAPIES TO TREAT AD HAVE BEEN LARGELY UNSUCCESSFUL. TWO IMPORTANT RISK FACTORS FOR AD, AGING, AND DIET, WITH AGING BEING THE GREATEST RISK FACTOR FOR AD, REMAIN LARGELY IGNORED. DIETARY RESTRICTION (DR), ONE OF THE MOST ROBUST INTERVENTIONS TO SLOW AGING, ALSO DELAYS THE ONSET OF ALZHEIMER’S DISEASE (AD) IN MULTIPLE MODELS ACROSS SPECIES. WE EXPLOITED THE SHORT LIFESPAN AND POWERFUL GENETIC TOOLS IN D. MELANOGASTER TO IDENTIFY THAT MUSTARD (MTD)/OXIDATIVE STRESS RESISTANCE PROTEIN 1 (OXR1) IN NEURONS IS REQUIRED FOR THE PROTECTIVE EFFECTS OF DR ON LIFESPAN. IMPORTANTLY, WE HAVE OBSERVED THAT OXR1 PROTECTS AGAINST AGE-RELATED NEURODEGENERATION IN FLY AND HUMAN-INDUCED PLURIPOTENT STEM CELL (IPSC) DERIVED MODELS OF NEURODEGENERATIVE DISEASES. THE MECHANISMS BY WHICH OXR1 PROTECTS AGAINST NEURODEGENERATION REMAINS UNCLEAR. WE OBSERVED THAT INHIBITING OXR1 REDUCES RETROMER PROTEINS WHILE ENHANCING RETROMER FUNCTION, RESCUES THE DELETERIOUS EFFECTS OF INHIBITING OXR1. FURTHERMORE, WE FOUND THAT ALTERATIONS IN OXR1 AND SEVERAL RETROMER PROTEINS ARE ASSOCIATED WITH AN INCREASED RISK OF AD IN HUMANS USING PROTEOMICS DATA FROM OVER 1000 AD PATIENTS FROM THE ACCELERATING MEDICINES PROGRAM-ALZHEIMER’S DISEASE (AMP-AD) NETWORK. HERE, WE PROPOSE TO TEST THE HYPOTHESIS THAT OXR1 ENHANCES RETROMER FUNCTION TO SLOW AGING AND NEURODEGENERATION USING FLY AND HUMAN IPSC MODELS OF AD. IN THE FIRST AIM, WE WILL DETERMINE THE MECHANISMS OF REGULATION OF OXR1 AND HOW THAT INFLUENCES AGING AND AGE-RELATED NEURONAL DAMAGE. IN THE SECOND AIM, WE WILL DETERMINE THE DR-DEPENDENT ROLE OF OXR1 IN ENHANCING RETROMER FUNCTION AND THE ROLE OF RETROMER IN MEDIATING THE PROTECTIVE EFFECTS OF DR. TO DETERMINE THE MECHANISM BY WHICH OXR1 ENHANCES RETROMER FUNCTION UPON DR; WE WILL USE PROTEOMICS TO DETERMINE AND CHARACTERIZE THE PROTEIN BINDING PARTNERS OF OXR1. IN THE THIRD AIM, WE WILL TEST THE ROLE OF OXR1 IN PROTECTING AGAINST NEURODEGENERATION IN MODELS OF AD, AND BY ENHANCING RETROMER FUNCTION. WE WILL TEST WHETHER OXR1 MODULATES AD PATHOLOGY IN FLY MODELS THAT OVEREXPRESS HUMAN TAU OR AMYLOID SS (ASS). WE WILL OVEREXPRESS OXR1 AND RETROMER PROTEINS IN FOREBRAIN CHOLINERGIC AND CORTICAL NEURON DERIVED AD IPSCS AND MEASURE AD ENDPOINTS: ASS42/40 ACCUMULATION, CELL DEATH, AND ELECTROPHYSIOLOGY. BECAUSE OXR1 REGULATES RETROMER FUNCTION IN THE FLY, WE WILL EVALUATE WHETHER THIS REGULATION IS CONSERVED IN HUMAN AD-DERIVED IPSCS AND CARRY OUT OMICS APPROACHES TO IDENTIFY KEY SIGNALING PATHWAYS MEDIATING OXR1’S PROTECTIVE EFFECTS. BY CHARACTERIZING RETROMER FUNCTION AND PROTEIN NETWORKS REGULATED BY OXR1 AND THEIR ROLE IN AGING AND AGE-DEPENDENT NEURODEGENERATION, WE WILL PROVIDE NOVEL TARGETS FOR DEVELOPING THERAPEUTICS TO SLOW AD-RELATED PATHOLOGIES AND EXTEND HEALTHSPAN. FURTHERMORE, WE WILL DETERMINE WHETHER REUSE OF PROTEINS THROUGH RETROMER UNDER NUTRIENT LIMITING CONDITIONS IS NEUROPROTECTIVE AND SLOWS AGING.

OXMRM: A TECHNIQUE TO QUANTIFY OXIDATION OF ENDOGENOUS REDOX-SENSITIVE CYSTEINES

THE ROLE OF NATURAL GENETIC VARIANTS IN NUTRIENT-DEPENDENT CHANGES IN HEALTHSPAN USING D. MELANOGASTER

ROLE OF OXR1 AND THE RETROMER IN BRAIN AGING AND ALZHEIMER'S DISEASE AND RELATED DEMENTIAS - PROJECT SUMMARY/ABSTRACT ALTHOUGH SOME GENETIC RISK FACTORS FOR AD ARE KNOWN, THE PRECISE ETIOLOGY OF MOST CASES OF LATE-ONSET ALZHEIMER'S DISEASE (AD) IS UNKNOWN. UNFORTUNATELY, AD THERAPIES HAVE BEEN LARGELY UNSUCCESSFUL. TWO IMPORTANT RISK FACTORS FOR AD, AGING, AND DIET, REMAIN POORLY UNDERSTOOD. DIETARY RESTRICTION (DR), ONE OF THE MOST ROBUST INTERVENTIONS TO SLOW AGING, ALSO DELAYS THE ONSET OF AD IN MULTIPLE MODELS ACROSS SPECIES. WE EXPLOITED THE SHORT LIFESPAN AND POWERFUL GENETIC TOOLS IN D. MELANOGASTER TO SHOW THAT MUSTARD (MTD)/OXIDATION RESISTANCE 1 (OXR1) IS ESSENTIAL FOR DR-MEDIATED LIFESPAN EXTENSION. IMPORTANTLY, MTD/OXR1 PROTECTS AGAINST AGE-RELATED NEURODEGENERATION IN FLY AND HUMAN-INDUCED PLURIPOTENT STEM CELL (IPSC)-DERIVED MODELS OF NEURODEGENERATIVE DISEASES. THE MECHANISMS BY WHICH MTD/OXR1 PROTECTS AGAINST NEURODEGENERATION REMAIN UNCLEAR. HOWEVER, WE OBSERVED THAT MTD/OXR1 DIRECTLY BINDS RETROMER PROTEINS, AND INHIBITING MTD REDUCES RETROMER PROTEINS. IMPORTANTLY, ENHANCING RETROMER FUNCTION MITIGATES THE HARMFUL EFFECTS OF MTD/OXR1 INHIBITION, REINFORCING THE HYPOTHESIS THAT RETROMER ACTS DOWNSTREAM OF OXR1. WE OBSREVED THAT OXR1 AND OTHER RETROMER PROTEINS DECLINE WITH AGE IN HUMANS AND ARE ASSOCIATED WITH AN INCREASED RISK OF AD IN HUMANS, USING PROTEOMICS DATA FROM OVER 2000 AD PATIENTS FROM THE ACCELERATING MEDICINES PROGRAM–ALZHEIMER’S DISEASE NETWORK. HERE, WE WILL TEST THE HYPOTHESIS THAT MTD/OXR1 ENHANCES RETROMER FUNCTION TO SLOW AGING AND NEURODEGENERATION IN FLY AND HUMAN IPSC MODELS OF AD. IN AIM 1, WE WILL DETERMINE THE ROLE OF SPECIFIC RETROMER COMPONENTS IN MEDIATING THE PROTECTIVE EFFECTS OF MTD/OXR1. WE WILL DETERMINE THE DOWNSTREAM MECHANISMS (E.G., ENDOCYTIC RECYCLING, SYNAPTIC HOMEOSTASIS, SPHINGOPLIPID, AND ALDEHYDE METABOLISM) BY WHICH MTD/OXR1 AND RETROMER INFLUENCE AGE-RELATED NEURODEGENERATION AND AGING. IN AIM 2, WE WILL DETERMINE THE ROLE OF SPECIFIC RETROMER COMPONENTS AND THE DOWNSTREAM TARGETS OF OXR1 IN FLY MODELS OF TAUOPATHY. THIS WILL TEST HOW MTD/OXR1 MODULATES AD PATHOLOGY IN FLY MODELS THAT OVEREXPRESS HUMAN TAU. IN AIM 3, WE WILL OVEREXPRESS OXR1 AND RETROMER PROTEINS IN FOREBRAIN CHOLINERGIC AND CORTICAL NEURON-DERIVED AD IPSCS AND MEASURE AD ENDPOINTS. WE WILL ALSO DETERMINE IF THE RETROMER MEDIATES THE PROTECTIVE EFFECTS OF MTD/OXR1 AND IF THIS REGULATION IS CONSERVED IN HUMAN MODELS OF AD. BY CHARACTERIZING RETROMER FUNCTION, PROTEIN NETWORKS REGULATED BY MTD/OXR1, AND THEIR ROLE IN AGING AND AGE-DEPENDENT NEURODEGENERATION, WE WILL PROVIDE NOVEL TARGETS FOR DEVELOPING THERAPEUTICS TO SLOW AD-RELATED PATHOLOGIES AND EXTEND HEALTHSPAN. WITH THE PROPOSED STUDY, WE WILL IDENTIFY THE CONSERVED MECHANISMS BY WHICH MTD/OXR1 MODULATES RETROMER AND SYNAPTIC HOMEOSTASIS TO INFLUENCE LIFESPAN AND ITS NEUROPROTECTIVE EFFECTS IN BOTH FLY AND HUMAN IPSC-DERIVED MODELS OF AD. THESE STUDIES WILL HAVE A SIGNIFICANT IMPACT ON UNDERSTANDING HOW LONGEVITY-RELATED PATHWAYS CAN BE TARGETED TO DEVELOP THERAPEUTICS FOR AD AND RELATED DEMENTIAS.

ENHANCING INDERDISCIPLINARY RESEARCH IN AGING AND AGE-RELATED DISEASE.

EVALUATING DIVERSE TECHNOLOGIES FOR DETECTING AND VALIDATING SENESCENT CELLS IN VIVO - PROJECT SUMMARY SINGLE CELL PROFILING HAS RECENTLY EXPLODED INTO MAINSTREAM BIOLOGY. SINGLE CELL PROFILING HAS BEEN EMPLOYED IN MYRIAD APPLICATIONS IN BIOLOGY, INCLUDING MULTIPLE DISEASES, EMBRYONIC DEVELOPMENT, COMPARATIVE EVOLUTIONARY STUDIES AND AGING. MOST RECENTLY, SINGLE CELL PROFILING HAS INFORMED THE PHENOMENA OF CELLULAR SENESCENCE. THIS PROPOSAL HAS TWO PHASES, UG3 (MODEL SYSTEMS), AND UH3 (HUMAN VALIDATION). CELLULAR SENESCENCE IS A MULTI-FACETED CELL FATE THAT ARRESTS CELL PROLIFERATION, ESSENTIALLY IRREVERSIBLY, AND ACTIVATES THE PRODUCTION AND SECRETION OF PRO-INFLAMMATORY CYTOKINES, CHEMOKINES, GROWTH FACTORS, PROTEASES AND LIPIDS, TERMED THE SENESCENCE ASSOCIATED SECRETORY PHENOTYPE (SASP). THE SASP CAN INFLUENCE TISSUE MICROENVIRONMENTS, AND THUS SENESCENT CELLS CAN STRONGLY AFFECT TISSUE FUNCTION AND LIKELY THE SYSTEMIC MILIEU. SENESCENT CELLS INCREASE WITH AGE AND CAN DRIVE A GROWING LIST OF AGE-RELATED PATHOLOGIES, RANGING FROM NEURODEGENERATION TO CANCER, IN PART THROUGH THE SASP. THERE IS INCREASING EVIDENCE THAT THERE ARE NO UNIVERSAL MARKERS FOR SENESCENT CELLS. INSTEAD, SENESCENT CELLS, WHILE SHARING CERTAIN CHARACTERISTICS AND BIOMARKERS, ARE REMARKABLY HETEROGENEOUS, VARYING IN CHARACTERISTICS WITH GENOTYPE, CELL AND TISSUE TYPE, SENESCENCE INDUCER, TISSUE (AND CELL CULTURE) MICROENVIRONMENT, AND CHRONOLOGY (TIME AFTER INITIAL SENESCENCE INDUCTION). WHILE SOME OF THE MORE COMMONLY EMPLOYED SENESCENCE MARKERS HAVE UTILITY IN SUPERFICIALLY IDENTIFYING SENESCENT CELLS DE NOVO, THE ONUS REMAINS ON THE INVESTIGATOR TO DEMONSTRATE WHY A CELL SHOULD BE CONSIDERED SENESCENT, RATHER THAN RELYING ON HISTORICAL MARKERS SUCH AS P16INK4A OR P21CIP1. THUS, NEW TECHNOLOGIES DESIGNED TO IDENTIFY NOVEL SENESCENT CELLS AND PHENOTYPES ARE NECESSARY THAT WILL REQUIRE VALIDATION BOTH IN CULTURE AND IN TISSUE. THE ULTIMATE GOAL OF THIS PROPOSAL IS TO DEVELOP NEW TECHNOLOGIES TO MAP SENESCENT SIGNATURES BACK TO INTACT HUMAN TISSUE. THIS GOAL WILL ENABLE US TO IDENTIFY AND SPATIALLY CHARACTERIZE SENESCENT CELLS IN EACH TISSUE, UNCOVERING UNIQUE MARKERS DEPENDING ON TISSUE AND CELL TYPE. PILOT 1: IDENTIFY SENESCENT CELLS IN THE MOUSE VASCULATURE, AND DETERMINE IF THEY CAN BE DETECTED BY ULTRASOUND, AND VERIFIED IN HUMAN TISSUE; PILOT 2: DEVELOP A MICROPHYSIOLOGIC EX VIVO TISSUE-ON-A-CHIP TO MODEL OVARIAN SENESCENCE, AND HUMAN TISSUE-TISSUE INTERACTIONS VIA THE SASP. THESE TWO PILOTS WILL USE A COMBINATION OF CELL SURFACE MARKERS IDENTIFIED IN OUR INITIAL PROFILING, AND DIGITAL SPATIAL PROFILING (DSP, NANOSTRING) OR VISIUM (10X) TO LOCALIZE SENESCENT SIGNATURES TO MORPHOLOGICAL STRUCTURES AND CELLS IN TISSUE SECTIONS. OUR PROPOSAL WILL DEVELOP VALIDATED MARKERS OF SENESCENCE IN MULTIPLE TISSUES AND CELL TYPES NOT PREVIOUSLY CHARACTERIZED TO DEPLOY THESE TECHNOLOGIES TO THE BROADER COMMUNITY.

IRON DYSREGULATION AND PARKINSON'S DISEASE

NEUROPEPTIDE MODULATION OF BEHAVIOR IN C. ELEGANS

EXOGENOUS KETONES TARGET GEROSCIENCE PATHWAYS AND AMELIORATE GERIATRIC SYNDROMES - THIS APPLICATION FOR THE NIA RESEARCH AND ENTREPRENEURIAL DEVELOPMENT IMMERSION AWARD (K01) DESCRIBES THE FIVE-YEAR CAREER DEVELOPMENT PLAN OF DR. BRIANNA STUBBS, A TRANSLATIONAL SCIENTIST AT THE BUCK INSTITUTE FOR RESEARCH ON AGING, NOVATO, CA. DR. STUBBS’ LONG-TERM CAREER GOAL IS TO BE AN INDEPENDENT CLINICAL RESEARCHER AND ENTREPRENEUR, AND TO BUILD A TRACK RECORD OF TRANSLATING ACADEMIC CONCEPTS INTO THERAPEUTICS TARGETING AGING AND AGE-RELATED DISEASE. THE SPECIFIC CAREER DEVELOPMENT GOALS OUTLINED IN THIS APPLICATION INCLUDE DEVELOPING EXPERTISE IN THE STUDY OF GEROSCIENCE BIOMARKERS; CLINICAL WORK WITH OLDER ADULTS, IND ENABLING STUDIES FOR DRUG DEVELOPMENT, AND TRANSLATIONAL ENTREPRENEURSHIP. DR. STUBBS WILL BE SUPPORTED BY A TEAM OF ACADEMIC, INDUSTRY AND BUSINESS DEVELOPMENT MENTORS TO ACCOMPLISH THESE CAREER DEVELOPMENT GOALS. THE CAREER DEVELOPMENT PLAN AND RESEARCH PLAN OF DR. STUBBS INCLUDES INDIVIDUALIZED MENTORSHIP WITH HER MENTORSHIP TEAM, FORMAL COURSEWORK, AND MEETINGS, AND IS BUILT AROUND IMMERSION IN THE BUCK BUSINESS DEVELOPMENT DEPARTMENT, EXPERIENCE IN AN ACADEMIC CLINICAL TRIAL OF OLDER ADULTS AND IN AN INDUSTRY DRUG DEVELOPMENT PROGRAM. DR. STUBBS WILL USE BIOSPECIMENS FROM THESE STUDIES IN HER RESEARCH PLAN. THE PLAN WILL COMPLEMENT AND EXTEND DR. STUBBS’ EXTENSIVE EXPERIENCE IN CLINICAL RESEARCH WITH YOUNG ADULTS, HUMAN PHYSIOLOGY, AND DEVELOPMENT OF KETONE-BASED CONSUMER FOOD PRODUCTS, TO EQUIP HER TO BE AN INDEPENDENT ENTREPRENEUR-SCIENTIST. THE OVERALL OBJECTIVE OF THE RESEARCH PLAN IS TO IDENTIFY GEROSCIENCE BIOMARKERS FOR USE IN FUTURE KETONE-BASED DRUG DEVELOPMENT. KETONES ARE PRODUCED FROM STORED FAT DURING FASTING OR STRENUOUS EXERCISE AND STRONG PRECLINICAL DATA DEMONSTRATES THAT THEY CAN IMPACT LIFESPAN AND HEALTHSPAN BY ACTING AS AN OXIDATIVE FUEL AND A SIGNALING METABOLITE. THE CENTRAL HYPOTHESIS OF THIS PROJECT IS THAT KETONE BODIES MODULATE GEROSCIENCE PATHWAYS COMMON TO GERIATRIC SYNDROMES INCLUDING INFLAMMATION, SENESCENCE, AND MITOCHONDRIAL HEALTH. THE SPECIFIC AIMS OF THE RESEARCH PLAN ARE TO INVESTIGATE THE EFFECTS OF EXOGENOUS KETONES ON THE SPECIFIC GEROSCIENCE MECHANISMS OF INFLAMMATION, SENESCENCE, AND MITOCHONDRIAL HEALTH IN A RODENT MODEL OF HEART FAILURE AND IN PRE-FRAIL AND FRAIL OLDER ADULTS, AND TO CORRELATE CHANGES IN THESE PATHWAYS TO FUNCTIONAL OUTCOMES. THE APPLICATION IS RELEVANT TO NIH AND NIA BECAUSE DR. STUBBS’ CAREER GOAL IS TO LEVERAGE AN UNDERSTANDING OF THE MULTIFACTORIAL PATHWAYS THAT REGULATE AGING AND LONGEVITY TO PROVIDE TRANSLATIONAL THERAPIES FOR THE MULTIFACTORIAL GERIATRIC SYNDROMES. THE ANTICIPATED RESULTS OF THE RESEARCH PLAN WILL ADVANCE THE PROGRESS OF KETONE-BASED TREATMENTS FOR HEART FAILURE. IF SUCCESSFUL, DR. STUBBS’ CAREER WILL RESULT IN THE DEVELOPMENT OF MULTIPLE INTERVENTIONS FOR GERIATRIC SYNDROMES, WHICH ULTIMATELY LEADS TO GREATER HEALTH AND INDEPENDENCE IN OLDER ADULTS.

REGULATION OF SYNAPTIC ACTIVITY BY MRNA METHYLATION - PROJECT SUMMARY NEUROTRANSMITTER RELEASE AT SYNAPTIC TERMINALS IS TIGHTLY CONTROLLED, AND ITS DYSREGULATION HAS PROFOUND CONSEQUENCES FOR NEURAL CIRCUIT ACTIVITY AND DISEASE. HERE, WE PROPOSE TO EXAMINE THE IN VIVO IMPACT OF N6- METHYLADENOSINE (M6A), THE MOST COMMON INTERNAL MODIFICATION OF MRNA, ON SYNAPTIC BIOLOGY. OUR ONGOING EFFORTS UNCOVER HOW DROSOPHILA MUTANTS OF M6A FACTORS ALTER NORMAL SYNAPTIC FUNCTION AT THE NEUROMUSCULAR JUNCTION (NMJ) AND SUGGEST NEW MECHANISMS FOR M6A REGULATION. WE WILL BUILD ON THIS KNOWLEDGE TO DISSECT HOW M6A CONTROLS THE ABILITY OF NEURONS TO ADJUST THEIR SET POINT FOR NEUROTRANSMITTER RELEASE AT THE SYNAPSE. THIS PROPOSAL WILL (1) ESTABLISH CRITICAL ROLES FOR THE EPITRANSCRIPTOME IN NORMAL SYNAPTIC FUNCTION, (2) PROVIDE COMPREHENSIVE GENOMIC DATA ON M6A TARGETS AND THE REGULATORY IMPACTS OF THESE MODIFICATIONS, AND (3) DEFINE NEW REGULATORY MECHANISMS BY WHICH M6A OPERATES AND INTERSECTS WITH OTHER NEURONAL REGULATORY PATHWAYS. THIS WORK WILL PROVIDE CONCEPTUAL ADVANCES TO OUR KNOWLEDGE OF THE REGULATION OF NEURONAL ACTIVITY UNDER PHYSIOLOGICAL CONDITIONS AND IN DISEASE.

SUMMER TRAINING COURSE IN EXPERIMENTAL AGING RESEARCH

MOLECULAR MECHANISMS OF LIFESPAN EXTENSION BY DIETARY RESTRICTION IN DROSOPHILA

LEICA TCS SP5 MP CONFOCAL AND TWO-PHOTON MICROSCOPE

TRIPLETOF 5600 HIGH RESOLUTION MASS SPECTROMETER

SYNAPTIC MECHANISMS OF TAU-MEDIATED PATHOGENESIS IN HUMAN IPSC-DERIVED NEURONS

CELLULAR SENESCENCE AND CONTROL OF GENE EXPRESSION

MTORC1 SIGNALING IN AGING AND METABOLISM

COMPREHENSIVE SYNAPTOME PROTEOMICS TARGETING PROTEIN EXPRESSION AND PTMS IN HD

SCREENING FOR APPNEO INHIBITORS

POTENTIAL ROLE OF LYSOSOMAL ATP13A2 IN CELLULAR IRON HOMEOSTASIS

ENVIRONMENTAL EXPOSURE AND ASTROCYTIC SENESCENCE: NOVEL LINK TO PD

ALTERNATIVE FOLDING OF SOLUBLE PROTEINS INDUCED BY A?42 AND TAU IN AGED CELLS - ABSTRACT ONE OF THE MAJOR GOALS OF ALZHEIMER’S DISEASE (AD) RESEARCH IS TO UNDERSTAND HOW AGING AND THE AGGREGATION OF ASS AND/OR HYPERPHOSPHORYLATED TAU DISRUPT THE PROTEOSTASIS AND CAUSE PROGRESSIVE NEURONAL DEATH. THE CURRENT PARADIGM OF AD RESEARCH FOCUSES ON THE AMYLOID AGGREGATION OF ASS/TAU AND THE PROTEINS SEQUESTRATED BY THESE AGGREGATES. A KEY UNTOUCHED QUESTION OF PROTEOSTASIS IS WHETHER THE FOLDING AND CONFORMATION OF SOLUBLE PROTEINS, WHICH REPRESENT >99% OF THE PROTEOME, ARE ALSO CHANGED DURING AGING AND AD. AS PROTEIN CONFORMATION DETERMINES THE SPECIFIC CELLULAR FUNCTION(S) OF A GIVEN PROTEIN, THE CHANGES IN PROTEIN FOLDING AND CONFORMATION WILL DISRUPT THE ORIGINAL PROTEIN FUNCTIONS OR ALLOW SOME POLYPEPTIDES TO ACQUIRE AGE-/AD-SPECIFIC FUNCTIONS. NASCENT POLYPEPTIDES TAKE KINETICALLY AND THERMODYNAMICALLY FAVORABLE ROUTES DURING DE NOVO FOLDING. THE ENERGY LANDSCAPE THAT DETERMINES THE FOLDING PATHS AND FINAL NATIVE CONFORMATIONS OF A PROTEIN IS SHAPED BY THE PHYSIOCHEMICAL ENVIRONMENT SURROUNDING THE NASCENT PEPTIDE. USING SACCHAROMYCES CEREVISIAE (BUDDING YEAST) AS A MODEL ORGANISM AND A NOVEL PROTEOMICS-STRUCTURE PIPELINE, OUR PRELIMINARY STUDIES REVEALED THAT CHANGING THE CELLULAR FOLDING ENVIRONMENT INDUCES THE ALTERNATIVE FOLDING OF MANY PROTEINS. AS CHANGES IN THE PROTEOME COMPOSITION AND METABOLIC PROFILE, TWO KEY COMPONENTS OF THE CELLULAR FOLDING ENVIRONMENT, ARE CONSERVED HALLMARKS OF AGING AND AD, THE GOAL OF THIS PROPOSAL IS TO UNDERSTAND HOW AGING AND AD AFFECT THE FOLDING AND CONFORMATION OF THE SOLUBLE PROTEINS. WE PROPOSE TO USE BUDDING YEAST FOR THIS PILOT PROJECT AS IT IS A COMMON MODEL ORGANISM USED TO STUDY CELLULAR AND MOLECULAR MECHANISMS OF AGING. YEAST SHARES NUMEROUS BIOLOGICAL PROCESSES WITH ANIMALS AND DEVELOPS AGE-RELATED CELLULAR DYSFUNCTIONS SIMILAR TO HUMAN AGING. IN ADDITION, THE YEAST MODEL OF AD HAS BEEN DEVELOPED TO REVEAL CONSERVED MECHANISMS OF AMYLOID AGGREGATION AND THE CELLULAR TOXICITY OF ASS AND TAU. AGING IS THE MAJOR RISK FACTOR OF AD AND OTHER AGE-RELATED DEMENTIAS. HOWEVER, PREVIOUS STUDIES LOOKED AT THE YOUNG YEAST CELLS EXPRESSING HUMAN ASS/TAU PROTEINS. IN THIS PROPOSED PROJECT, WE WANT TO COMBINE THE STRENGTH OF AGING AND AD RESEARCH IN YEAST TO FIND ALTERNATIVE PROTEIN FOLDING INDUCED BY ASS42/TAU IN AGED CELLS. WE INTEND TO ACCOMPLISH OUR GOAL BY 1) DETERMINING THE PREVALENCE OF PROTEIN ALTERNATIVE FOLDING INDUCED BY AGING, AND 2) DISSECTING THE EFFECT OF ASS42/TAU ON PROTEIN FOLDING IN AGED CELLS. THE HITS FROM OUR PROJECT WILL BE A RESOURCE FOR RESEARCHERS IN THE FIELD TO DETERMINE THE GAIN OR LOSS OF PROTEIN FUNCTIONS ASSOCIATED WITH THESE ALTERNATIVE FOLDING OF PROTEINS. THIS WORK IS EXPECTED TO REVEAL THE FIRST 3D VIEW OF THE PROTEOME AFFECTED BY AGING AND ASS42/TAU.

CELL COMPETITION AS A NOVEL ASPECT OF CELLULAR SENESCENCE DURING AGING

GENOME-WIDE ANALYSIS OF NON-CODING RNA FROM RETROTRANSPOSONS IN HUMAN ADULT STEM

TARGETING CONSERVED DIET-RESPONSIVE TRANSCRIPTIONAL NETWORKS IN NEURONS TO SLOW NEURODEGENERATION IN ALZHEIMER'S DISEASE - PROJECT SUMMARY/ ABSTRACT ALZHEIMER’S DISEASE (AD) IS A CHRONIC NEURODEGENERATIVE DISORDER AND THE MOST COMMON CAUSE OF DEMENTIA AMONGST SUBJECTS OVER THE AGE OF 65. DESPITE IDENTIFYING SOME OF THE GENETIC RISK FACTORS FOR AD, THERAPEUTICS TO TREAT AD HAVE BEEN UNSUCCESSFUL. A POTENTIAL REASON FOR THIS IS THAT AGING, THE MOST SIGNIFICANT RISK FACTOR FOR AD, HAS NOT BEEN CONSIDERED. DIETARY RESTRICTION (DR) IS ONE OF THE MOST ROBUST INTERVENTIONS TO SLOW AGING AND THE ONSET OF AGE-RELATED DISEASES, INCLUDING AD. HOWEVER, THE UNDERLYING MECHANISMS BY WHICH DR PROTECTS AGAINST AD ARE UNKNOWN. WE PROPOSE TO USE D. MELANOGASTER TO INVESTIGATE THE CONSERVED LINKS BETWEEN DIET AND AD FOR THE FOLLOWING REASONS: 1) THEIR EXCELLENT TRACK RECORD FOR ELUCIDATING THE BIOLOGY OF AGING, NEURODEGENRATION AND DR, 2) THEIR AMENABILITY TO GENETIC MANIPULATION, 3) THE AVAILABILITY OF ESTABLISHED GENETIC MODELS FOR UNDERSTANDING AGING AND AD PATHOLOGY, 4) THEIR FAST GENERATION TIME AND SHORT LIFESPAN, AND 5) THEY SHARE MANY BIOLOGICAL PROCESSES AND SIGNALING PATHWAYS WITH MAMMALS. THE OVERALL GOALS OF THIS PROPOSAL ARE TO UNDERSTAND THE MECHANISMS BY WHICH DR INFLUENCES AD PATHOLOGY AND NEURODEGENERATION. WE HAVE OBSERVED THAT DR SIGNIFICANTLY IMPROVES SURVIVAL AND REDUCES THE FUNCTIONAL DECLINE IN TAUOPATHY MODELS OF AD IN FLIES. IMPORTANTLY, OUR PROTEOMIC ANALYSIS SUGGESTS THAT MUTANT TAU AFFECTS THE PROTEOME OF FLIES IN A FASHION THAT IS SIMILAR TO THE EFFECT OF A HIGH-NUTRIENT DIET. A COMPARISON OF OUR PROTEOMIC ANALYSIS FROM A FLY TAUOPATHY MODEL WITH PROTEOMIC DATA FROM HUMAN AD BRAINS IDENTIFIED 47 COMMON GENES. WE ALSO DEMONSTRATE THAT MODULATING TWO OF THE TRANSCRIPTIONAL REGULATORS AND THEIR DOWNSTREAM GENES MODULATE NEURODEGENERATION IN TAU MUTANT FLIES. BASED ON OUR PRELIMINARY DATA, OUR CENTRAL HYPOTHESIS IS THAT NUTRIENT-DEPENDENT TRANSCRIPTIONAL NETWORKS IN THE BRAIN INFLUENCE NEURODEGENERATION IN PATHOGENIC TAU AND AD MODELS. WE WILL TEST OUR HYPOTHESIS BY PURSUING THE FOLLOWING SPECIFIC AIMS. IN AIM 1, WE CHARACTERIZE THE TRANSCRIPTIONAL TARGETS ALTERED IN BOTH FLY TAUOPATHY MODEL AND HUMAN AD FOR CHANGES IN NEURODEGENERATION AND LIFESPAN IN TAU AND ASS FLY MODELS OF AD. IN AIM 2, WE CHARACTERIZE CANDIDATE TRANSCRIPTIONAL REGULATORS FOR CHANGES IN NEURODEGENERATION AND LIFESPAN IN FLY MODELS OF AD. IN AIM 3, WE IDENTIFY THE MECHANISMS BY WHICH NUTRIENT-RESPONSIVE TRANSCRIPTIONAL NETWORKS INHIBIT NEURODEGENERATION. WE FOCUS ON DIET'S IMPACT ON METABOLISM, OXIDATIVE STRESS, HETEROCHROMATIN LOSS, AND ABORTIVE NEURONAL CELL-CYCLE ACTIVATION, GIVEN THAT THESE PROCESSES ARE KNOWN TO AFFECT AD AND AGING. THIS RESEARCH IS SIGNIFICANT, AS WE EXPECT IT TO REVEAL COMMON GENETIC MECHANISMS ACROSS SPECIES, NOVEL TARGETS, AND LIFESTYLE CHANGES THAT SLOW THE ONSET AND PROGRESSION OF AD AND RELATED TAUOPATHY.

THE ROLE OF ADVANCED GLYCATION END PRODUCTS IN MODULATING HEALTHSPAN USING C. ELEGANS

A MODEL OF KIDNEY STONE DISEASE USING D. MELANOGASTER

VITAMIN D METABOLISM AND LIFESPAN DETERMINATION

METAL HOMEOSTASIS AND AGING

APP SIGNALING NETWORK

LYSINE MALONYLATION AND SIRT5 IN EPIGENETIC REGULATION

AB SCIEX QTRAP 5500 LC MS/MS SYSTEM

ADVANCED GLYCATION END PRODUCTS AS NON-AUTONOMOUS FACTORS THAT DRIVE AGING

COPAS FP-PRO 500 FLOW CYTOMETER

MECHANISMS OF RRAS REGULATION OF HUNTINGTIN AND TURNOVER

EFFECT OF APOE ISOFORMS ON ALZHEIMER'S DISEASE AND AGING USING ISOGENIC HUMAN APOE IPSC MODEL

CELLULAR SENESCENCE AND ALZHEIMER'S DISEASE - PROJECT SUMMARY / ABSTRACT SENESCENT CELLS DEVELOP A SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP) INVOLVING PRO-INFLAMMATORY AND PRO-OXIDATIVE FACTORS THAT CAN ELICIT DELETERIOUS PARACRINE-LIKE EFFECTS ON NEIGHBORING CELLS. INDEPENDENTLY OF THE ORIGINAL STRESSOR, SENESCENCE CAN ALSO SPREAD FROM SENESCENT TO NON-SENESCENT BYSTANDER CELLS IN A PROCESS KNOWN AS SENESCENCE-INDUCED SENESCENCE (SIS). NEURONS WERE HISTORICALLY CONSIDERED TO BE UNABLE TO UNDERGO CELLULAR SENESCENCE. RECENT RESEARCH HAS HOWEVER PROVIDED EVIDENCE THAT NEURONS MAY BE ABLE TO UNDERGO SENESCENCE DURING NORMAL AGING AND DISEASE. SENOLYTIC DRUGS, WHICH SELECTIVELY KILL SENESCENT CELLS, HAVE FOR EXAMPLE BEEN SHOWN TO IMPROVE FUNCTIONS IN THE BRAINS OF ANIMAL MODELS OF ALZHEIMER’S DISEASE (AD) AND RELATED DEMENTIAS. THIS NEW APPROACH IS OF GREAT IMPORTANCE—CLINICAL TRIALS INVESTING EFFICACY OF AMYLOID BETA (AΒ) ANTIBODIES HAVE LED TO DISAPPOINTING FAILURES AND NO KNOWN DISEASE-MODIFYING TREATMENTS HAVE TO DATE BEEN IDENTIFIED. THERE IS HOWEVER A CAVEAT TO THE USE OF SENOLYTICS FOR AD AND RELATED DISORDERS. IT IS NOT KNOWN WHETHER SENOLYTICS KILL SENESCENT GLIAL CELLS AND SPARE NEURONS BECAUSE THEY CANNOT SENESCE, OR IF SENOLYTICS NOT ONLY KILL SENESCENT GLIAL CELLS BUT ALSO SENESCENT NEURONS. SENESCENT CELLS IN THE PERIPHERY ARE NORMALLY REMOVED BY IMMUNE CELLS INCLUDING NATURAL KILLER (NK) CELLS. DUE TO ITS UNIQUE IMMUNE PRIVILEGE, SENESCENT CELLS IN THE BRAIN MAY INITIALLY BE ABLE TO EVADE REMOVAL BY NK CELLS, ALLOWING FOR AN INCREASE AND SPREAD IN THE OVERALL SENESCENT CELL LOAD. WHEN IMMUNE PRIVILEGE BECOMES COMPROMISED IN LATER STAGES OF THE DISEASE, THIS COULD RESULT IN A LARGE-SCALE REMOVAL OF SENESCENT CELLS INCLUDING NEURONS. IT IS UNCLEAR WHETHER THIS WOULD BE BENEFICIAL OR DETRIMENTAL TO GLOBAL BRAIN HEALTH. WE WILL INITIALLY ASSESS NEURONAL SENESCENCE IN MIXED PRIMARY HUMAN NEURONAL-ASTROCYTIC 2D AND 3D CULTURES RECENTLY GENERATED BY OUR LABORATORY IN RESPONSE TO ABETA-MEDIATED STRESS. WE WILL THEN DETERMINE THE ROLE OF SENESCENCE IN VIVO UTILIZING A WELL-CHARACTERIZED AD MOUSE MODEL, THE 3XTG AD LINE. THIS LINE HAS BEEN CROSSED WITH A P16-3MR TRANSGENIC MOUSE MODEL THAT ALLOWS THE IDENTIFICATION AND INDUCIBLE ABLATION OF SENESCENT CELLS. USE OF THESE MODELS WILL ALLOW US TO DETERMINE: (1) THE ABILITY OF NEURONS, ASTROCYTES, AND OTHER CNS CELL TYPES TO UNDERGO STRESS-MEDIATED SENESCENCE AND A SASP, (2) WHETHER THIS IS ACCOMPANIED BY AN ABETA-INDEPENDENT SIS, (3) WHETHER LATE-STAGE LOSSES IN IMMUNE PRIVILEGE ENABLES CLEARANCE OF SENESCENT CELLS VIA INFILTRATION OF PERIPHERAL IMMUNE CELLS, AND (4) WHETHER SENESCENT CELL REMOVAL IS BENEFICIAL OR DETRIMENTAL AND AT WHAT STAGE OF DISEASE PROGRESSION.

SENESCENT CELLS AS A SOURCE OF PRO-GERONIC FACTORS

POLYRIBOSOME TARGETS MEDIATING MRNA DECAY FOR CANCER PREDICTION AND THERAPY

ANALYSIS OF GENE EXPRESSION AND CELL FUNCTION IN SINGLE CELL CORTICAL OSTEOBLASTS

SPATIAL PROTEOMICS OF OSTEOARTHRITIS IN BONES TO UNCOVER DISEASE PROGRESSION - PROJECT SUMMARY / ABSTRACT THERE IS AN URGENT NEED TO DEVELOP NOVEL AND INNOVATIVE TECHNOLOGIES TO ADVANCE HEALTH AND THERAPEUTIC INTERVENTIONS AND TO DEVELOP TOOLS FOR MONITORING OF DISEASE PROGRESSION IN THE MUSCULOSKELETAL SYSTEM. MOLECULAR DETAILS OF DISEASE AND INJURY OF THE MUSCULOSKELETAL SYSTEM ARE PARTICULARLY NEEDED FOR NOVEL PROTEOMIC ASSAYS TO MONITOR DISEASE PROGRESSION IN OSTEOARTHRITIS OR MENISCAL INJURY. OUR INNOVATIVE APPROACH INCLUDES DEVELOPMENT OF MASS SPECTROMETRY IMAGING (MSI) TECHNOLOGIES FOR JOINTS TO SPATIALLY VISUALIZE PROTEINS OF THE EXTRACELLULAR MATRIX (ECM) AND GLYCANS, AND TO APPLY DEEP PROTEOMIC PROFILING OF BONES. THESE TECHNOLOGICALLY NOVEL APPROACHES WILL ALSO ENABLE US TO DEVELOP URGENTLY NEEDED MOLECULAR ASSAYS TO ASSESS THE EFFICACY OF INTERVENTIONS. MOLECULAR CHANGES RELATED TO AGE, INJURY OR OSTEOARTHRITIS (OA) IN MUSCULOSKELETAL TISSUES, PARTICULARLY BONE AND CARTILAGE, ARE TYPICALLY SPATIALLY DISTINCT. THIS IS EVEN MORE EVIDENT WHEN CONSIDERING TEMPORAL AND DYNAMIC CHANGES DURING EMERGING FUNCTIONAL DECLINE. OUR NATIONAL TEAM PROVIDES A UNIQUE COMBINATION OF EXPERTISE TO DEVELOP NEW, INNOVATIVE AND PARADIGM-CHANGING TOOLS TO INVESTIGATE BONE DISEASES, SUCH AS OA, AGE-RELATED CHANGES IN BONE, AND CHANGES INDUCED BY INJURY TO MENISCUS. DR. SCHILLING (BUCK INSTITUTE) IS AN EXPERT IN PROTEOMICS, GLOBAL BONE ANALYSIS, SENESCENCE AND AGING, DR. ALLISTON (UC SAN FRANCISCO) IS AN EXPERT IN ORTHOPEDICS, BONE MECHANO-BIOLOGY, AND OSTEOARTHRITIS DISEASE BIOLOGY, AND DR. ANGEL (MEDICAL UNIVERSITY OF SOUTH CAROLINA, MUSC) IS AN EXPERT IN MSI, SPATIAL ECM PROTEOMICS, AND GLYCAN BIOLOGY. WE WILL EMPLOY AND OPTIMIZE STATE-OF-THE-ART MASS SPECTROMETRY IMAGING (MSI) TECHNOLOGIES TO SPATIALLY RESOLVE MOLECULAR CHANGES IN TISSUES (USING A MOLECULAR READOUT MASS/CHARGE: M/Z) IN HUMAN AND MOUSE TISSUE SLICES MONITORING HUNDREDS OF ANALYTES SIMULTANEOUSLY. THESE INNOVATIVE TECHNOLOGIES INDICATE PREMISE TO DEVELOP INTO GROUNDBREAKING TOOLS FOR THE ORTHOPEDIC AND MUSCULOSKELETAL COMMUNITY TO INVESTIGATE BONE AND CARTILAGE DECLINE, AND TO MONITOR INTERVENTIVE TREATMENTS. WITH THIS PARADIGM CHANGE, WE WILL BE ABLE TO IMAGE HUNDREDS OR EVEN THOUSANDS OF MOLECULES SIMULTANEOUSLY AT 15 TO 40 ΜM SPATIAL RESOLUTION. PROTEIN SIGNATURES FROM THE EXTRACELLULAR MATRIX IN HUMAN OA KNEE TISSUES REVEALED VERY ADVANCED DISEASE PROGRESSION IN CARTILAGE AND BONE AT THE INNER (MEDIAL) SIDE OF THE KNEE (WITH HEALTHIER TISSUE AT THE OUTER/LATERAL SIDE) DISCOVERING DISTINCT NOVEL OA PROTEIN TARGETS. THESE RESULTS AND NOVEL SPATIAL ASSAYS, IN COMBINATION WITH DEEP PROTEOMIC PROFILING OF THE BONE PROTEOME (>2,000 PROTEIN GROUPS), WILL PROVIDE INSIGHTS INTO DISEASE MECHANISMS AND ALLOW FOR UNBIASED PROTEOMIC MONITORING DURING FUNCTIONAL DECLINE OF JOINTS AND BONES. ROBUST STATISTICAL PROCESSING IS IMPLEMENTED. AS INDIVIDUAL LIFESPANS INCREASE ACROSS THE WORLD, AGE-RELATED COMPLICATIONS ARE INCREASING AND DEVELOP INTO A MAJOR PUBLIC HEALTH CHALLENGE. THE UNIQUE MS IMAGING AND PROTEOME PROFILING CAPABILITIES OF OUR TEAM PROVIDE MAJOR OPPORTUNITIES FOR THE DISCOVERY OF NOVEL DISEASE TARGETS FOR THERAPEUTIC INTERVENTIONS.

CELLULAR SENESCENCE, AGING AND CANCER DEVELOPMENT (PQ7)

CELLULAR SENESCENCE AS A MEDIATOR OF MITOCHONDRIAL DYSFUNCTION-INDUCED AGING

INVESTIGATING THE INTERPLAY BETWEEN HALLMARKS OF AGING; PROTEIN GLYCATION, NUTRIENT SENSING, AND SENESCENCE - PROJECT SUMMARY CELLULAR SENESCENCE AND DEREGULATED NUTRIENT SENSING (INSULIN SIGNALING AND/OR INSULIN RESISTANCE), TWO HALLMARKS OF AGING, ARE IMPLICATED IN THE LOSS OF PHYSIOLOGICAL FUNCTION AND ETIOLOGY OF AGE-RELATED DISEASES. ADVANCED GLYCATION END PRODUCTS (AGES) ARE A SOURCE OF INCREASED PROTEIN GLYCATION BURDEN, A MANIFESTATION OF ANOTHER HALLMARK OF AGING, LOSS OF PROTEOSTASIS. METHYLGLYOXAL (MGO), A REACTIVE PRECURSOR TO AGES, INFLUENCES THE PATHOPHYSIOLOGY OF INSULIN RESISTANCE (IR) AND CELLULAR SENESCENCE. A RECENT STUDY INDICATES THAT IR ACCELERATES SS-CELL SENESCENCE LEADING TO FUNCTIONAL DECLINE IN PANCREATIC TISSUE AND WORSENING METABOLIC PROFILE. EMERGING EVIDENCE ALSO SUGGESTS THAT THE ACCUMULATION OF SENESCENT CELLS IN ADIPOSE TISSUE DIRECTLY REGULATES THE IR PHENOTYPES. CURRENT INVESTIGATIONS ESTABLISHED THE CO-OCCURRENCE OF THESE TWO AGING HALLMARKS. HOWEVER, A MAJOR GAP IN OUR KNOWLEDGE EXISTS IN UNDERSTANDING THE INTERACTIONS BETWEEN BOTH THE HALLMARKS AND THE IMPACT OF NORMAL AND PATHOLOGICAL AGING ON THESE INTERACTIONS. OUR LONG-TERM GOAL IS TO FURTHER INVESTIGATE THIS EMERGING AND EXCITING AREA OF RESEARCH IN THE CONTEXT OF TISSUE-SPECIFIC COMMUNICATION THAT DICTATES INTERACTIONS AMONG AGING HALLMARKS. THE PRESENT PROPOSAL WILL SPECIFICALLY ASK THE FOLLOWING CRITICAL QUESTIONS: (A) HOW DO THESE THREE HALLMARKS OF AGING INTERACT WITH EACH OTHER; (B) IS THERE ANY TISSUE-SPECIFIC HIERARCHY AND INTERDEPENDENCY BETWEEN THE THREE HALLMARKS IN DIFFERENT TISSUES; (C) CAN MULTIPLE INTERVENTIONS WHICH OVERCOME THESE HALLMARKS OF AGING SHOW ADDITIVE PROTECTION TO SLOW AGING AND AGE-RELATED DISEASES. OUR CENTRAL HYPOTHESIS IS THAT MGO- INDUCED PROTEIN GLYCATION NOT ONLY ACTS AS AN ENDOGENOUS DRIVER FOR IR AND CELLULAR SENESCENCE BUT ALSO PLAYS A KEY ROLE IN THE INTERPLAY BETWEEN THESE TWO HALLMARKS IN THE ADIPOSE TISSUE AND PANCREAS. WE WILL TEST THE HYPOTHESIS BY PURSUING THE FOLLOWING SPECIFIC AIMS: 1) THE ROLE OF METHYLGLYOXAL IN INDUCING SENESCENCE IN THE PANCREATIC SS-CELLS AND ADIPOSE TISSUE THROUGH PROTEIN GLYCATION; 2) THE ROLE OF METHYLGLYOXAL IN INDUCING IR IN ADIPOSE TISSUE OR INSULIN RELEASE FROM PANCREATIC SS-CELLS THROUGH PROTEIN GLYCATION; AND 3) INVESTIGATE THE INTERPLAY BETWEEN MGO-INDUCED PROTEIN GLYCATION IR AND CELLULAR SENESCENCE. PREADIPOCYTES AND PRIMARY SS-CELLS WILL BE USED TO STUDY THE ROLE OF MGO IN INDUCING TISSUE-SPECIFIC SENESCENCE AND IR. WE WILL ALSO UTILIZE SENOLYTICS TO DETERMINE THE IMPORTANCE OF SENESCENT CELLS IN MGO-INDUCED IR. WE WILL ALSO DETERMINE THE THERAPEUTIC POTENTIAL OF COMBING A NOVEL GLYCATION-LOWERING COCKTAIL, GLY-LOW, WITH OR WITHOUT A SENOLYTIC FOR THEIR IMPACT ON AMELIORATING IR, SENESCENCE, AND LIFESPAN IN A PHYSIOLOGICAL (HIGH-FAT DIET) MODEL. A KEY SIGNIFICANCE OF THIS WORK IS HELPING US UNDERSTAND THE INTERCONNECTIVITY AMONG MGO-INDUCED PROTEIN GLYCATION BURDEN, IR, AND CELLULAR SENESCENCE, THEREBY SHEDDING LIGHT ON HOW THESE HALLMARKS INTERACT WITH EACH OTHER UNDER STRESS CONDITIONS AND HOW THIS KNOWLEDGE CAN BE USED TO SLOW AGING AND AMELIORATE AGE-RELATED DISEASES.

THE ROLE OF CANNABINOID SIGNALING IN THE NEMATODE CAENORHABDITIS ELEGANS

ROLE OF GSH AND MAO-B IN PD-RELATED MITO DYSFUNCTION

POSTDOCTORAL REEARCH TRAINING AND EDUCATION IN GEROSCIENCE (10 OF 11) TL1

TOXICITY MECHANISMS OF A?42 AND TAU IN AGED CELLS - ABSTRACT ALZHEIMER'S DISEASE (AD) IS CHARACTERIZED BY A PROGRESSIVE LOSS OF NEURONAL STRUCTURES AND FUNCTIONS THAT UNDERLIE COGNITIVE DEFECTS AND DEMENTIA. THE NEURONAL DAMAGE HAS BEEN LINKED TO THE AGGREGATION OF ASS PEPTIDES AND HYPERPHOSPHORYLATED TAU PROTEINS. HOWEVER, CLINICAL RESULTS OF REDUCING THESE NEUROTOXIC PROTEINS HAVE BEEN DISAPPOINTING. THESE FAILURES SUGGEST THAT AD IS A MULTIFACTORIAL PROCESS AND THAT MANY CELLULAR DEFECTS WERE LEFT UNCORRECTED AFTER REMOVING THE ASS/TAU DEPOSITIONS. FINDING THESE RESISTANT CELLULAR DEFECTS IS A CRITICAL STEP TOWARDS THE SUCCESSFUL INTERVENTION OF AD. STUDIES IN HUMANS HAVE SHOWN THAT AGING IS THE BIGGEST RISK FACTOR FOR AD. HOWEVER, MOST CELLULAR MECHANISMS OF ASS/TAU TOXICITY ARE GENERATED BY RESEARCH IN YOUNG CELLS. AS AGED CELLS HAVE DIFFERENT PROTEOMES AND PHYSIOLOGY THAN YOUNG CELLS, STUDYING THE CELLULAR TOXICITY OF ASS42/TAU IN AGED CELLS IS A BETTER MODEL FOR THEIR TOXICITY IN OLDER AD PATIENTS AND IS REQUIRED TO REVEAL THE RESISTANT CELLULAR DEFECTS THAT ARE INDUCED BY ASS42/TAU DURING AGING. THE GOAL OF THIS PROPOSAL IS TO DETERMINE HOW ASS/TAU, TOGETHER WITH AGING, CHANGES THE CELLULAR COMPARTMENTALIZATION AND WHETHER REJUVENATING SOME OF THESE DEFECTS CAN PROTECT CELLS FROM THE TOXICITY OF ASS/TAU. IDENTIFYING THESE AGE-SPECIFIC ASS/TAU SENSITIVE TARGETS WILL REVEAL NOVEL MECHANISMS OF ASS/TAU TOXICITY AND ENABLE STRATEGIES TO RESCUE CELLULAR DEFECTS THAT ARE LEFT UNCORRECTED AFTER REMOVING THE ASS/TAU DEPOSITIONS. WE PROPOSE TO USE SACCHAROMYCES CEREVISIAE (YEAST) AND DROSOPHILA MELANOGASTER (FLY) FOR THIS PILOT PROJECT AS THESE TWO MODEL ORGANISMS ARE WIDELY USED TO STUDY MOLECULAR MECHANISMS OF AGING AND AD. YEAST AND FLY SHARE MANY BIOLOGICAL PROCESSES WITH HUMANS AND DEVELOP AGE- RELATED CELLULAR DYSFUNCTIONS SIMILAR TO HUMAN AGING. THE YEAST MODEL OF AD HAS BEEN USED TO REVEAL CONSERVED MECHANISMS OF AMYLOID AGGREGATION AND THE CELLULAR TOXICITY OF ASS AND TAU, AS WELL AS THE GENETIC AND CHEMICAL TOXICITY MODIFIERS. HOWEVER, THESE STUDIES LOOKED AT THE YOUNG YEAST CELLS EXPRESSING THE HUMAN ASS/TAU PROTEINS. THEREFORE, WE WANT TO COMBINE THE STRENGTH OF AGING AND AD RESEARCH IN YEAST TO SCREEN FOR THE DEFECTS OF PROTEIN EXPRESSION AND LOCALIZATION CAUSED BY ASS42/TAU IN AGED CELLS. OUR STRATEGY IS TO USE AGED YEAST TO SCREEN FOR HITS AND TEST THE CONSERVED ONES IN FLY MODELS OF AD. WE INTEND TO ACCOMPLISH OUR GOAL BY 1) DETERMINING HOW ASS AND TAU PROTEINS INTERFERE WITH PROTEIN EXPRESSION AND LOCALIZATION IN REPLICATIVELY OLD YEAST CELLS, AND 2) EVALUATING THE BENEFICIAL EFFECT OF NEWLY IDENTIFIED LONGEVITY FACTORS IN FLY MODELS OF AD. THE YEAST SCREEN WILL BE ACCOMPLISHED BY USING A NEW HIGH-THROUGHPUT MICROSCOPIC SCREENING METHOD DEVELOPED IN OUR LAB FOR AGED CELLS. THE INSIGHTS GAINED FROM THE YEAST SCREEN WILL ALSO BE TESTED IN THE FLY MODELS OF AD. THIS WORK WILL ADVANCE OUR UNDERSTANDING OF THE INTERACTIONS BETWEEN AGING AND AD-RELATED PROTEINS THAT TOGETHER CAUSE THE AGE-RELATED NEURODEGENERATIVE DISEASES.

IMPACT OF KIBRA SIGNALING ON MEMORY IN TAUOPATHY

IDENTIFICATION OF BLOOD BIOMARKERS PREDICTIVE OF ORGAN AGING - PROJECT SUMMARY AS WE AGE, OUR TISSUES AND ORGANS EXPERIENCE MOLECULAR AND PHYSIOLOGICAL DAMAGE THAT PREVENTS THEM FROM FUNCTIONING PROPERLY AND THIS ULTIMATELY LEADS TO DISEASE STATES. THESE CHANGES ARE NOT ONLY DUE TO THE AGING PROCESS ITSELF BUT ARE LARGELY INFLUENCED BY THE EXPOSOME WHICH INCLUDES ALL NON-GENETIC EXPOSURES (ENVIRONMENTAL AND BEHAVIORAL). DEPENDING ON THE COMPLEX INTERACTION BETWEEN THE EXPOSOME OF AN INDIVIDUAL AND THEIR GENETICS, DIFFERENT ORGANS DETERIORATE OVER TIME AT A DIFFERENT PACE, RESULTING IN TISSUES WITH DIFFERENT BIOLOGICAL AGES WITHIN THE SAME INDIVIDUAL. AS THE BIOLOGICAL AGE OF A GIVEN ORGAN REFLECTS ITS OVERALL HEALTH AND FUNCTIONAL CAPACITY, BIOLOGICALLY OLDER ORGANS ARE MORE LIKELY TO CAUSE HEALTH PROBLEMS INCREASING THE RISK OF DISEASES. AGING “CLOCKS” POWERED BY OMICS TECHNOLOGIES (TRANSCRIPTOMICS, PROTEOMICS, EPIGENOMICS, ETC.) AND MACHINE LEARNING METHODS HAVE BEEN USED TO APPROXIMATE THE BIOLOGICAL AGE OF SPECIFIC TISSUES. HOWEVER, TISSUE-SPECIFIC CLOCKS REQUIRE OMICS DATA FROM A BIOPSY, MAKING CLINICAL ADOPTION IMPRACTICAL. THEREFORE, THERE IS A CRITICAL NEED TO DEVELOP SIMPLE DIAGNOSTIC TOOLS USING READILY ACCESSIBLE BIOLOGICAL MATERIAL TO MEASURE ORGAN- SPECIFIC AGING RATES IN AN INDIVIDUAL WHICH CAN BE TRANSLATED INTO PERSONALIZED ACTIONABILITIES AND ENABLE ACCURATE EVALUATION OF THE EFFICACY OF HEALTH-PROMOTING INTERVENTIONS. USING BLOOD, THE PIPELINE OF THE IMMUNE SYSTEM, FROM AGING COHORTS WE AND OTHERS HAVE DEMONSTRATED THAT ACCELERATED AGING, AS EVIDENCED BY AGE-RELATED CHRONIC INFLAMMATION (INFLAMMAGING) AND DYSFUNCTIONAL IMMUNE SYSTEMS, RESULTS IN ORGAN DYSFUNCTION AND AN ELEVATED RISK OF DISEASE IN OLDER SUBJECTS. THIS IS NOT SURPRISING SINCE INFLAMMAGING HAS BEEN PROPOSED TO BE A COMMON DENOMINATOR OF MOST, IF NOT ALL, DISEASES OF AGING. IN THIS PROPOSAL, WE HYPOTHESIZE THAT THE BIOLOGICAL INFORMATION TO INVESTIGATE THE AGING RATES OF A GIVEN ORGAN IS CONTAINED IN THE BLOOD OF THE SAME INDIVIDUAL AND THUS, CAN BE ESTIMATED USING A COLLECTION OF TISSUE-SPECIFIC GENE EXPRESSION SIGNATURES MATCHED WITH THOSE FROM BLOOD SAMPLES. HERE, WE WILL ASSEMBLE MULTIPLE PUBLIC DOMAIN DATASETS WITHIN AND OUTSIDE OF THE NIH COMMON FUND TO CREATE BLOOD-BASED ORGAN-SPECIFIC CLOCKS AND ENABLE RAPID DIAGNOSTICS OF AGING RATES FOR A GIVEN ORGAN IN AN INDIVIDUAL. TO DO SO, WE WILL USE TRANSCRIPTOMIC DATA ACROSS MULTIPLE TISSUES AND MATCHED BLOOD FROM THE GENOTYPE-TISSUE EXPRESSION (GTEX) DATABASE TO CONSTRUCT A COMPUTATIONAL FRAMEWORK THAT CALCULATES THE RATE OF AGING OF 45 TISSUES IN AN INDIVIDUAL USING BLOOD GENE EXPRESSION. WE WILL VALIDATE THE RESULTING MODELS TO PREDICT ORGAN-SPECIFIC AGING IN DISEASE STATES SPECIFIC TO THE ORGAN OF INTEREST, AND WE WILL ASSESS THE INFLUENCE OF LIFESTYLE FACTORS INCLUDING DIET, EXERCISE AND SMOKING ON THE AGING OF DIFFERENT ORGANS USING DATA FROM THE FRAMINGHAM HEART STUDY. FINALLY, WE WILL USE THE LIBRARY OF INTEGRATED NETWORK-BASED CELLULAR SIGNATURES (LINCS) TO IDENTIFY CANDIDATE COMPOUNDS THAT CAN RESTORE THE GENE EXPRESSION CHANGES IN THE BLOOD ASSOCIATED WITH TISSUE AGING TO OPTIMAL LEVELS.

POSTDOCTORAL REEARCH TRAINING AND EDUCATION IN GEROSCIENCE (10 OF 11) RL9

TARGETING INFLAMMAGING WITH MESENCEPHALIC ASTROCYTE-DERIVED NEUROTROPHIC FACTOR (MANF).

ROLE OF S6 KINASE 2 IN AGING

SCREENING POTASSIUM AND PHOSPHATE BINDER DRUGS FOR LIFESPAN AND HEALTHSPAN EFFECTS IN INVERTEBRATES

ROLE OF MITOCHONDRIA IN ESTROGEN ACTION AND CELLULAR SENESCENCE

LIFESPAN EXTENSION BY DIFFERENTIAL TRANSLATION MEDIATED BY EIF-4G IN C. ELEGANS

ROLE OF CELLULAR SENESCENCE IN CARCINOGENESIS

4E-BP DEPENDENT SECRETED PROTEINS FROM HUMAN AND FLY SKELETAL MUSCLE

GENETIC DISSECTION OF NATURAL VARIATION IN CALORIC RESTRICTION-INDUCED CELLULAR L

ROLE OF MTOR PATHWAY IN CELLULAR SENESCENCE

FUNCTIONAL MAPPING OF THIRST CIRCUITS IN THE AGING MOUSE BRAIN

QUANTITATING, TARGETING, AND PHENOTYPING IN VIVO CELLULAR SENESCENCE IN HUMANS

THE REPRODUCTIVE AGING CONFERENCE - PROJECT SUMMARY WE ARE REQUESTING FUNDS TO SUPPORT THE REPRODUCTIVE AGING CONFERENCE ANNUALLY FOR 3 YEARS, 2023 - 2025. THE 2023 MEETING WILL BE HELD AT THE OMNI RANCHO LAS PALMAS RESORT (PALM SPRINGS, CALIFORNIA, USA) ON MAY 2 - 5, 2023. THIS WILL BE THE SECOND MEETING FOLLOWING THE INAUGURAL CONFERENCE IN JUNE 2022. AGING IN THE GONAD DRAMATICALLY AFFECTS AGING IN SOMATIC TISSUES, YET WE KNOW LITTLE ABOUT THE MECHANISMS REGULATING HEALTHY AGING IN REPRODUCTIVE ORGANS, NOR HOW THIS CROSSTALK IS ACHIEVED. THE REPRODUCTIVE AGING CONFERENCE WILL FOCUS ON UNDERSTANDING MECHANISMS THAT REGULATE AGING IN REPRODUCTIVE TISSUES AND THEIR RELATIONSHIP TO OVERALL ORGANISMAL HEALTHSPAN AND LONGEVITY. FUNDS ARE REQUESTED EACH YEAR TO HELP COVER EXPENSES OF 1) INVITED SPEAKERS AND 2) MERITORIOUS JUNIOR SCIENTISTS (STUDENTS, POSTDOCTORAL FELLOWS AND JUNIOR FACULTY) AND MINORITY TRAINEES WHO WILL BENEFIT FROM FORMAL AND INFORMAL INTERACTIONS WITH SENIOR SCIENTISTS, TO PRESENT THEIR RESEARCH FINDINGS, RECEIVE FEEDBACK FROM EXPERTS IN THE FIELD, AND EXCHANGE IDEAS FOR SHAPING THE FIELD. TO OUR KNOWLEDGE, THIS IS THE ONLY INTERNATIONAL MEETING DEDICATED ENTIRELY TO THE TOPIC OF REPRODUCTIVE AGING. THE GOAL OF THE REPRODUCTIVE AGING CONFERENCE IS TO SHOWCASE RESEARCH AND STIMULATE COLLABORATIONS THAT BRIDGE THE DISCIPLINES OF REPRODUCTIVE BIOLOGY AND AGING RESEARCH TO HELP DEFINE NEW PARADIGMS TO ACCELERATE PROGRESS. OUR PRIMARY OBJECTIVE IS TO BECOME THE PREMIER FORUM FOR COALESCING THE FIELD AND FOSTERING THE DISCUSSION OF CONCEPTS AND PRESENTATION OF RESEARCH AT THE FOREFRONT OF DISCOVERY RELATED TO AGING IN THE REPRODUCTIVE SYSTEM AND ITS CONNECTION TO AGING IN THE REST OF THE BODY. REPRODUCTIVE AGING RESEARCH ENCOMPASSES MULTIPLE FRONTS OF INTEREST: IDENTIFICATION OF THE MOLECULAR AND CELLULAR MECHANISMS THAT REGULATE AGING IN REPRODUCTIVE ORGANS THAT FACILITATE OVERALL HEALTHSPAN AND LONGEVITY; MECHANISMS OF GERM CELL AND NICHE AGING AND HOW THESE MIGHT REGULATE SYSTEMIC AGING; AND DEVELOPING NEW MODELS TO STUDY THESE INTERACTIONS. THE INTERSECTION OF THESE HISTORICALLY SEPARATE AREAS, THE BASIC BIOLOGY OF AGING AND AGING IN THE REPRODUCTIVE TRACT, HAVE BEGUN TO MERGE AROUND COMMON MOLECULAR PRINCIPLES. INVITED SPEAKERS INCLUDE ESTABLISHED AS WELL AS OUTSTANDING EARLY CAREER SCIENTISTS SELECTED FOR THE QUALITY OF THEIR SCIENCE AND ABILITY TO STIMULATE DEBATE. THIS CONFERENCE WILL PROVIDE A FORUM FOR BOTH FORMAL AND INFORMAL INTERACTIONS BETWEEN TRAINEES AND FACULTY AT ALL STAGES TO PROMOTE THE EXCHANGE OF IDEAS AND COLLABORATIVE SYNERGIES AROUND REPRODUCTIVE AGING. POSTER AND ORAL PRESENTATIONS WILL EXAMINE REPRODUCTIVE AGING AT THE MOLECULAR, CELLULAR AND ORGANISMAL LEVEL, INCLUDING ITS MOLECULAR DRIVERS, AND THE EFFECTS OF GENETIC AND ENVIRONMENTAL FACTORS ON RELEVANT CELLULAR/SYSTEM PROPERTIES AND INTERACTIONS ACROSS DIVERSE MODEL SYSTEMS RANGING FROM WORMS TO HUMANS. THIS UNIQUE CONFERENCE WILL BRING TOGETHER SCIENTISTS TACKLING THIS PROBLEM FROM FUNDAMENTAL TO CLINICAL ANGLES TO BRAINSTORM CREATIVE WAYS TO BUILD AND STRENGTHEN THE FIELD.

MODULATING ALZHEIMER'S DISEASE PROGRESSION BY PRESERVING INTESTINAL HEALTH.

CONTROL OF STEM CELL FUNCTION AND TISSUE REGENERATION BY THE CRL4 COMPLEX

ALTERED MTOR SIGNALING ASSOCIATED WITH LMNA MUTATIONS.

CHAPERONE-MEDIATED AUTOPHAGY IN SPINOCEREBELLAR ATAXIA TYPE 7 (SCA7).

HUNTINGTON'S DISEASE: ANALYSIS OF PROTEOLYSIS

INVESTIGATING THE ROLE OF LAMINB IN INFLAMMATION-ASSOCIATED STEM CELL DYSFUNCTION DURING AGING

CONSERVED ROLE OF URIC ACID AND PURINE METABOLITES IN LONGEVITY AND HEALTHSPAN

IDENTIFYING THE PATHWAYS BY WHICH NATURALLY-OCCURRING GENETIC VARIANTS ASSOCIATED WITH NUTRIENT RESPONSE REGULATE LONGEVITY AND HEALTH.

A NEW PROTEOSTASIS CHECKPOINT IN DROSOPHILA INTESTINAL STEM CELLS: IMPLICATIONS FOR AGE-RELATED TISSUE DYSFUNCTION

THE ROLE OF SODIUM AND ZINC IN A DROSOPHILA MODEL OF NEPHROLITHIASIS

AGE-DEPENDENT MANGANESE TOXICITY IN PARKIN MUTANTS

CELL FATE DIVERSITY IN AGING

INVESTIGATING FATTY ACID OXIDATION IN PRIMARY AND METASTATIC MYC-OVEREXPRESSING TRIPLE-NEGATIVE BREAST CANCER

US HUPO: NOVEL PROTEOMIC PERSPECTIVES ON CANCER, AGING AND DISEASE

ELUCIDATING THE ROLE OF TGF-BETA SIGNALING IN HUNTINGTON'S DISEASE