Winifred Masterson Burke Medical Research

PHASE II RANDOMIZED CONTROLLED TRIAL OF BENFOTIAMINE IN EARLY ALZHEIMER'S DISEASE - WE PROPOSE A SEAMLESS PHASE 2A-2B TRIAL INVESTIGATING BENFOTIAMINE, A PRODRUG OF THIAMINE, AS A FIRST-IN- CLASS SMALL MOLECULE TREATMENT FOR EARLY ALZHEIMER'S DISEASE (AD). WE CALL THIS PROPOSED TRIAL `BENFOTIAMINE IN EARLY ALZHEIMER'S DISEASE (BEAD)”. BRAIN TISSUE THIAMINE DEFICIENCY CAUSES MEMORY DEFICITS THAT ARE REVERSIBLE WITH THIAMINE TREATMENT IN PRECLINICAL AD MODELS AND IN HUMAN CONDITIONS INCLUDING WERNICKE KORSAKOFF SYNDROME. IN ANIMAL MODELS OF MILD IMPAIRMENT OF OXIDATIVE METABOLISM (I.E., THIAMINE DEFICIENCY), NEURONAL LOSS IS ACCOMPANIED BY CHANGES IN NEUROFILAMENT LIGHT (NFL), BY INCREASED NEUROINFLAMMATION (GLIAL FIBRILLARY ACID PROTEIN GFAP), BY ELEVATION OF ADVANCED GLYCATION END PRODUCTS (AGE) AND BY INCREASED PLAQUE AND BY TANGLE PATHOLOGY, ALL OF WHICH OCCUR IN AD. BENFOTIAMINE DRAMATICALLY RAISES BLOOD AND BRAIN TISSUE THIAMINE IN THESE MODELS, CONFERRING BEHAVIORAL BENEFIT AND REDUCED PLAQUE AND TANGLE FORMATION. WE PREVIOUSLY CONDUCTED AN EARLY PHASE 2 PILOT SINGLE-SITE 12-MONTH DOUBLE BLIND PLACEBO CONTROLLED RCT OF 600 MG OF BENFOTIAMINE IN 71 PERSONS WITH EARLY AD. BENFOTIAMINE WAS WELL TOLERATED, HAD ENCOURAGING PHARMACOKINETIC (PK) AND PHARMACODYNAMIC (PD) RESPONSES AND SHOWED BENEFITS ON THE CLINICAL DEMENTIA RATING (CDR), THE ADAS-COG, AND MARKERS OF BRAIN METABOLISM. THERE IS NEW EVIDENCE IN MICE THAT A 1200 MG OF BENFOTIAMINE FURTHER INCREASES THIAMINE LEVELS WITH GREATER COGNITIVE BENEFITS. THUS, WE ARE PROPOSING AN 18-MONTH PHASE 2A/2B RANDOMIZED PLACEBO CONTROLLED RCT OF BENFOTIAMINE TESTING 600 MG/DAY AND 1200 MG/DAY IN 400 PARTICIPANTS EARLY AD, INCLUDING MILD COGNITIVE IMPAIRMENT (MCI) AND MILD DEMENTIA WITH PLASMA EVIDENCE OF AMYLOID POSITIVITY. OUR OVERARCHING HYPOTHESIS IS THAT SIGNIFICANT BENEFITS IN COGNITION AND GLOBAL FUNCTION WILL OCCUR WITH DOSES OF BENFOTIAMINE THAT ARE SAFE, WELL TOLERATED, AND ACHIEVE SUFFICIENT TARGET ENGAGEMENT. IF THIS PHASE 2 TRIAL IS SUCCESSFUL WE HAVE A CONSOLIDATED SEAMLESS PHASE 3 DEVELOPMENT PLAN TO EXPEDITE BENFOTIAMINE REACHING PATIENTS. WE WILL TEST OUR OVERARCHING HYPOTHESIS THROUGH THE FOLLOWING AIMS:(1) TO EFFICIENTLY DETERMINE THE HIGHEST SAFE AND WELL-TOLERATED DOSE OF BENFOTIAMINE IN PHASE 2A (600 MG OR 1200 MG) THAT CAN BE ADVANCED TO LONG TERM CLINICAL ENDPOINTS AT 72 WEEKS; (2) TO EVALUATE THE EFFICACY OF BENFOTIAMINE IN PHASE 2B, TO BENEFIT (A) GLOBAL FUNCTION MEASURED WITH THE CDR SUM OF BOXES (CDR-SB) AND (B) COGNITION MEASURED WITH ADAS-COG13 DURING A TREATMENT PERIOD OF 72 WEEKS IN EARLY AD; (3) TO EVALUATE THE PK (SERUM THIAMINE AND IT'S ESTERS) AND PD EFFECTS (THIAMINE PYROPHOSPHATE ACTIVATION OF TRANSKETOLASE AND ADVANCED GLYCATION END-PRODUCTS (AGES)) OF BENFOTIAMINE, AND THEIR RELATION TO THE PRIMARY OUTCOMES; (4) TO EVALUATE THE DOWNSTREAM BIOLOGICAL EFFECTS OF TREATMENT WITH BENFOTIAMINE IN EARLY AD ON MEASURES OF NEURODEGENERATION (CORTICAL THICKNESS ON MRI, PLASMA NEUROFILAMENT LIGHT AND TOTAL TAU), NEUROINFLAMMATION (GLIAL FIBRILLARY ACID PROTEIN) AND AD PATHOPHYSIOLOGY INCLUDING P-TAU 231, AND ASS 42/40 RATIO.

MITOCHONDRIAL DYSFUNCTION IN NEUROGEGENERATION OF AGING

NEURAL PREDICTORS OF HAND THERAPY EFFICACY IN CHILDREN WITH CEREBRAL PALSY

PLASTICITY IN THE AGING OLFACTORY SYSTEM

SYNAPSE ELIMINATION IN THE CENTRAL NERVOUS SYSTEM

ELECTROPHYSIOLOGICAL EVALUATION OF BRAIN REGIONS VULNERABLE TO ALZHEIMERS DISEASE

MECHANISTIC LINKS BETWEEN THE BENEFITS OF PHARMACOLOGICALLY HIGH THIAMINE (VITAMIN B1) IN ALZHEIMER'S DISEASE TO ADVANCED GLYCATION ENDPRODUCTS (AGE) - EXPERIMENTAL DATA LINKING THIAMINE (VITAMIN B1) DEFICIENCY TO ALZHEIMER’S DISEASE (AD) INSPIRED OUR CLINICAL TRIAL, WHICH GENERATED PRELIMINARY EVIDENCE THAT PHARMACOLOGICAL THIAMINE PRODUCED BY THE DRUG BENFOTIAMINE PROVIDES CLINICAL BENEFIT. WE HYPOTHESIZE THAT PHARMACOLOGICAL THIAMINE IS PROTECTIVE BY DIMINISHING THE FORMATION OF ADVANCED GLYCATION ENDPRODUCTS (AGE). AGE ARE PROTEINS AND LIPIDS THAT BECOME GLYCATED AND HARMFUL FOLLOWING EXPOSURE TO REDUCING SUGARS. AGE CAUSE IRREVERSIBLE DAMAGE TO BIOLOGICAL MACROMOLECULES BY ALTERING THEIR STRUCTURAL AND FUNCTIONAL INTEGRITY. ABUNDANT EVIDENCE LINKS AGE TO AD. IN AD ANIMAL MODELS, THIAMINE DEFICIENCY INCREASES AGE AND EXACERBATES PLAQUES AND TANGLE FORMATION, WHILE INCREASED THIAMINE DIMINISHES AGE AND PATHOLOGY. IN OUR PILOT CLINICAL TRIAL, PHARMACOLOGICAL THIAMINE LEVELS DIMINISHED GLOBAL PLASMA AGE LEVELS AND IMPROVED SYMPTOMS IN PATIENTS WITH AD. INTERESTINGLY, IN AD PATIENTS, THE EFFECTS OF HIGH THIAMINE ARE DIMINISHED IN PATIENTS CARRYING THE APOE4 GENOTYPE, THE MOST SIGNIFICANT GENETIC RISK FACTOR FOR SPORADIC AD. WE POSTULATE THAT THIS IS BECAUSE APOE4 INCREASES UNIQUE AGE AT EARLIER STAGES. OPTIMIZING THIS THERAPEUTIC APPROACH REQUIRES A BETTER UNDERSTANDING OF THE MECHANISM UNDERLYING THE ACTION OF BENFOTIAMINE. ALL PREVIOUS RELATED AD AND THIAMINE STUDIES HAVE UTILIZED AGE ANTIBODY SURVEYS. THIS DATA IS LIMITED TO A SMALL RANGE OF AGE AND PROVIDES NO DATA ON THE PROTEINS AND SPECIFIC SITES MODIFIED WITH GLYCATION. WE WILL PROVIDE THIS CRITICAL DATA BY USING MULTIPLE STATE-OF-THE-ART MASS SPECTROMETRIC MEASURES OF AGE. GLOBAL GLYCAPROTEOMICS WILL IDENTIFY GLYCATED PROTEINS AND SPECIFIC SITES OF AGE MODIFICATIONS AND AGE- OMICS WILL IDENTIFY A BROAD RANGE OF CROSSLINKING AND NON-CROSSLINKING AGE. A SECOND MAJOR GAP IS THE LACK OF OUR UNDERSTANDING ON HOW APOE4 MODIFIES THE RESPONSE TO THIAMINE. NOVEL APOE3 AND APOE4 HUMANIZED APP MOUSE MODELS WILL ALLOW US TO TEST THESE INTERACTIONS. WE WILL TEST OUR HYPOTHESES: (1) IN AD AUTOPSY BRAINS AT DIFFERENT STAGES OF THE DISEASE, AGE MODIFICATIONS ARE CRITICAL TO THE PATHOPHYSIOLOGY OF AD IN AN APOE-DEPENDENT MANNER. (2) IN MOUSE MODELS OF AD, THIAMINE DEFICIENCY DRIVES AD-LIKE PATHOLOGY AND MEMORY LOSS BY CAUSING SPECIFIC BRAIN AND BLOOD AGE MODIFICATIONS WHICH ARE MODIFIED BY APOE GENOTYPE. (3) IN MOUSE MODELS, BENFOTIAMINE IS BENEFICIAL BY DIMINISHING SPECIFIC AGE AND THE TREATMENT MUST BE INITIATED AT AN EARLIER STAGE OF DISEASE IN APOE4 MICE. THESE STUDIES WILL DRAMATICALLY IMPROVE OUR UNDERSTANDING OF THE ROLE OF AGE IN AD AND ITS LINK TO A TREATMENT OF PHARMACOLOGICAL THIAMINE LEVELS. DEFINING THE INTERACTION OF AGE AND THIAMINE IN THE ETIOLOGY AND PROGRESSION OF AD WILL ENABLE THE DEVELOPMENT OF SPECIFIC AGE SIGNATURES FOR TARGETS OF ENGAGEMENT FOR THERAPEUTIC TRIALS AND AS AD DIAGNOSTIC AND PROGNOSTIC BIOMARKERS. THESE STUDIES WILL DEFINE THE DIFFERENTIAL EFFECTIVENESS OF THIAMINE BY APOE GENOTYPE AND DEFINE THE MOST EFFECTIVE THERAPEUTIC APPROACH FOR APOE3 AND CONSISTENTLY HARD TO TREAT APOE4 CARRIERS. WE THEREFORE EXPECT THIS STUDY WILL HAVE A HIGH IMPACT ON TRANSLATIONAL AD MEDICINE.

THE ROLE OF CD36 IN ISCHEMIC INFLAMMATION AND INJURY

CORTICOSPINAL NEURON DYSFUNCTION AND DEGENERATION IN ALS: TESTING THE ROLE OF CORTICOMOTOR CONNECTIVITY IN MOTOR NEURON DISEASE - ABSTRACT (SUMMARY): IN PATIENTS WITH AMYOTROPHIC LATERAL SCLEROSIS (ALS) AND THE RELATED MOTOR NEURON DISEASE (MND) PRIMARY LATERAL SCLEROSIS (PLS), DEFICITS IN MOTOR CONTROL OCCUR AS A CONSEQUENCE OF THE DEGENERATION OF CORTICOSPINAL NEURONS (CSNS). ALS IS MORE COMMON THAN PLS, AND GENETICALLY MORE COMPLEX, WITH FAMILIAL FORMS ASSOCIATED WITH CAUSAL MUTATIONS IN OVER 30 ALS-RELATED GENES. IN THESE ALS MICE, HOWEVER, DYSFUNCTION AND DEGENERATION OF CSNS HAVE NOT BEEN CAREFULLY EXAMINED, AND DATA IMPLICATING CORTICOSPINAL (CS) CIRCUITS IN THESE MODEL SYSTEMS OF ALS IS SURPRISINGLY LIMITED. ONE REASON FOR THIS MAY BE THE VERY DIFFERENT PATTERN OF CONNECTIVITY BETWEEN CSNS AND SPINAL MNS IN HUMANS VS. MICE. IN HUMANS, CS AXONS LOCATED IN THE VENTRAL AND LATERAL FUNICULI FORM DIRECT CONNECTIONS WITH BOTH MNS (CORTICO-MOTONEURONAL (CM) CONNECTIONS) AND INTERNEURONS. IN CONTRAST, CS AXONS IN MICE ARE LOCATED MAINLY IN THE DORSAL FUNICULUS AND ONLY FORM INDIRECT CONNECTIONS WITH MNS THROUGH PRE-MOTOR INTERNEURONS. THEREFORE, WE WILL USE PLEXINA1 MUTANT MICE WHICH HAVE CM CONNECTIONS TOGETHER WITH ALS MOUSE MODELS TO ANALYZE CS CIRCUITS. OUR CENTRAL HYPOTHESIS IS THAT PROGRESSIVE DEFECTS IN CS CIRCUITRY IN ALS MICE WILL BE EXACERBATED BY THE ESTABLISHMENT OF CM CONNECTIONS. IN AIM 1, WE WILL DETERMINE FORMATION OF CS CIRCUITS IN ALS MOUSE MODELS WITH CM CONNECTIONS. IN AIM 2, WE WILL DETERMINE FUNCTION OF CS CIRCUITS IN ALS MOUSE MODELS WITH CM CONNECTIONS. IN AIM 3, WE WILL EXAMINE SKILLED MOVEMENTS IN ALS MOUSE MODELS WITH CM CONNECTIONS. THESE STUDIES WILL PROVIDE A MODEL SYSTEM TO STUDY MECHANISMS OF CS DEGENERATION IN ALS/PLS, AND TO TEST NOVEL THERAPEUTICS TARGETING UPPER MOTOR NEURON DYSFUNCTION IN THESE DISORDERS.

TRANSCRANIAL DIRECT CURRENT STIMULATION AND ROBOTIC TRAINING IN CHRONIC STROKE

ENGAGING NEURON-INTRINSIC SIGNALING FOR AXON GROWTH AFTER SPINAL CORD INJURY

GRADING VISUAL IMPAIRMENT IN CHILDREN WITH BRAIN INJURY

DISSECTING SPINAL INTERNEURON CIRCUITS TO CONTROL SKILLED MOVEMENTS

MODULATION OF CORTICAL NETWORKS, A NEW APPROACH TO SPINAL CORD INJURY REHABILITATION

IMMUNE-MEDIATED MECHANISMS UNDERLYING CONDITIONING-INDUCED STROKE RECOVERY

VASCULAR CONTROL IN DIABETIC RETINOPATHY

B-RAF DRIVES REGENERATIVE AXON GROWTH IN THE OPTIC NERVE IN VIVO

RETINAL NEURAL PROCESSING DURING RETINAL DEGENERATIVE DISEASES

OPTICAL DISSECTION OF INTRACORTICAL CIRCUITS SUPPORTING MOTOR RECOVERY AFTER SPINAL CORD INJURY

TRANSCRIPTIONAL REGULATION OVER NEUROGENESIS OF CORTICAL OUTPUT NEURON SEGMENTAL IDENTITY AND DIVERSITY - SUBCEREBRAL PROJECTION NEURONS (SCPN) RESIDE IN THE NEOCORTEX, AND EXTEND AXONS TO SUBCEREBRAL TARGETS IN THE BRAINSTEM AND SPINAL CORD. CORTICOSPINAL NEURONS (CSN), A SUBCLASS OF SCPN, PROJECT TO THE SPINAL CORD AND THEIR AXONS FORM THE CORTICOSPINAL TRACT (CST), A CRITICAL CIRCUIT FOR VOLUNTARY MOTOR CONTROL. IN ADDITION, PROJECTIONS FROM THE NEOCORTEX TO BRAINSTEM TARGETS FUNCTION IN PARALLEL WITH CSN TO EXERT MOTOR CONTROL. DEGENERATION OF THESE PROJECTION NEURONS IN AMYOTROPHIC LATERAL SCLEROSIS (ALS), ALONG WITH DEGENERATION OF SPINAL MOTOR NEURONS, CAUSES SPASTICITY AND PARALYSIS. IN HUMANS, DAMAGE TO THE CST AFTER SPINAL CORD INJURY IS A PRINCIPAL CAUSE OF LOSS OF VOLUNTARY MOTOR CONTROL. FURTHER, INTEGRITY OF CORTICOSPINAL CONNECTIVITY IS CENTRALLY LINKED TO RECOVERY FROM STROKE AND CEREBRAL PALSY. THERE HAVE BEEN MULTIPLE INVESTIGATIONS DETAILING THE ROLE OF CORTICO-BRAINSTEM VS. CORTICOSPINAL PROJECTIONS IN BOTH MOTOR CONTROL, AS WELL AS THEIR DISTINCT CONTRIBUTIONS TO FUNCTIONAL RECOVERY IN THESE DISTINCT CAUSES OF PARALYSIS. HOWEVER, IT REMAINS UNCLEAR WHEN AND HOW THESE DISTINCT PROJECTIONS ARE ESTABLISHED DURING DEVELOPMENT. UNDERSTANDING THE MOLECULAR BASIS OF THIS SPECIFICATION AND DIFFERENTIATION DURING DEVELOPMENT THEREFORE HOLDS SIGNIFICANT PROMISE IN ESTABLISHING APPROACHES THAT ARE TAILORED TO ENHANCING PLASTICITY OF THESE RELATED, YET DISTINCT CIRCUITS. A NECESSARY FIRST STEP TOWARD THIS ULTIMATE GOAL IS TO IDENTIFY THE MOLECULAR MECHANISMS DIRECTING SCPN AXONS TO BRAINSTEM (CORTICO-BRAINSTEM NEURONS) VERSUS SPINAL CORD (CSN). ONGOING WORK IN OUR LAB HAS IDENTIFIED THAT SUCH PROJECTIONS ARE INITIALLY SPECIFIED DURING THE PROCESS OF AXON EXTENSION DURING DEVELOPMENT. WE HAVE IDENTIFIED THAT CORTICO-BRAINSTEM AND CORTICOSPINAL NEURONS EXPRESS DISTINCT GENES AND CAN BE MOLECULARLY DISTINGUISHED IN MICE BY BIRTH. FURTHER OUR DATA SUGGEST THAT THE TRANSCRIPTIONAL REGULATOR SATB2 ACTS, IN PART, TO SPECIFY CORTICO-BRAINSTEM NEURONS. THIS PROPOSAL INVESTIGATES THE HYPOTHESIS THAT TRANSCRIPTIONAL REGULATION CONTROLS THE DEVELOPMENT OF CORTICO-BRAINSTEM VS. CORTICOSPINAL PROJECTIONS BY LATE EMBRYONIC DEVELOPMENT IN MICE. BUILDING ON THIS FOUNDATION, WE WILL FIRST IDENTIFY THE TIME POINT IN DEVELOPMENT WHEN THESE PROJECTIONS ARE SPECIFIED USING KNOCK-IN CRE REPORTER MICE (AIM 1). THIS WILL BE TESTED USING INTRACEREBRAL INJECTIONS OF AAV-REPORTERS AT DISTINCT DEVELOPMENTAL TIMES IN UTERO WITH ADULT ANALYSIS OF AXONAL PROJECTIONS. IN AIM 2, WE WILL INVESTIGATE THE TRANSCRIPTIONAL TARGETS OF SATB2 IN SCPN AT LATER DEVELOPMENTAL TIMES BY PROFILING ALL SCPN IN SATB2 WT AND CONDITIONAL KO MICE, AS WELL AS BY SATB2 OVEREXPRESSION, AT SINGLE CELL RESOLUTION. FINALLY, IN AIM 3 USING SUBPOPULATION-SPECIFIC TRANSGENIC CRE LINES, WE WILL INVESTIGATE SCPN AXON TARGETING IN BOTH SATB2 LOSS- AND GAIN-OF-FUNCTION. IN ADDITION, WE WILL INVESTIGATE WHETHER MISEXPRESSION OF SATB2 TARGET GENES CAN ALTER SCPN TARGETING TO THE BRAINSTEM VS. SPINAL TARGETS. TOGETHER, OUR WORK WILL DISCERN IN-DEPTH, THE MECHANISMS OF WHEN AND HOW TRANSCRIPTIONAL REGULATION CONTROLS SCPN SEGMENTAL “IDENTITY” THEREBY PROVIDING A MECHANISTIC FRAMEWORK FOR SUBSEQUENT IDENTIFICATION OF MOLECULES CONTROLLING SEGMENTALLY APPROPRIATE SCPN CONNECTIVITY WITH SUBCEREBRAL TARGETS.

ROLE OF CD36 IN FUNCTIONAL RECOVERY IN CHRONIC STROKE

THE ROLE OF NEURONAL HYPEREXCITABILITY AND PROTEOSTASIS IN ALZHEIMER'S DISEASE - KEY PRODROMAL EVENTS IN ALZHEIMER’S DISEASE (AD) REVOLVE ON ALTERED ELECTRICAL SIGNALS AND BUILDUP OF GARBAGE PROTEINS IN VULNERABLE AREAS BEFORE SIGNS OF AD (LOCUS COERULEUS, LC)) AND AS DISEASE PROGRESSES TO BRAIN REGIONS THAT GOVERN MEMORY (HIPPOCAMPUS, HPC) AS DISEASE SEVERITY INCREASES. THIS STUDY BRIDGES THE POTENTIAL LIFESPAN OF DISEASE PROGRESSION USING A MOUSE MODEL OF AD PATHOLOGY TO EXAMINE HOW MODIFICATIONS IN BRAIN ACTIVITY CAN LEAD TO DISEASE. THE LC, WHILE NOT TRADITIONALLY ASSOCIATED WITH AD, IS THE SITE OF SOME OF THE EARLIEST PATHOLOGY IN AD, AS EARLY AS PEOPLE IN THEIR 20S. IT IS ALSO AN AREA REGULATING FLIGHT OR FIGHT RESPONSE AND AROUSAL. IN AD, THE HYPEREXCITATION OR UNREGULATED AROUSAL (HYPERAROUSAL) MAY BE A KEY EVENT IN MOVING THE DISEASE FROM AREAS LIKE THE LC, OR ANOTHER AREA WITH EARLY PATHOLOGY, THE ENTORHINAL CORTEX (EC) TO THE HIPPOCAMPUS OR OTHER CORTICAL REGIONS THAT ARE MORE COMMONLY ASSOCIATED WITH ALZHEIMER’S. IN AD, THE ACCUMULATION OF AGGREGATED PROTEINS DUE TO ALTERED CELLULAR PROCESSES, SPECIFICALLY, THE REGULATION OF PROTEIN LIFE CYCLE (PROTEOSTASIS) AND DYSFUNCTION OF AUTOPHAGIC-LYSOSOMAL AND UBIQUITIN-PROTEASOMAL SYSTEMS IS A KEY FEATURE OF NEUROPATHOLOGY. THESE PROCESSES ARE RESPONSIBLE FOR CLEARING THE GARBAGE IN CELLS AND DECLINES WITH AGE AND IS ACCELERATED IN DISEASE. WHILE WE KNOW LOSS OF PROTEOSTASIS CAN IMPAIR CELLULAR FUNCTION, HOW IT CAN IMPEDE NEURONAL ACTIVITY HAS NOT BEEN WELL ADVANCED. IN THIS APPLICATION, WE PROPOSE THAT A PRIMARY EVENT EARLY ON IS THE ALTERATION OF NEURAL NETWORKS IN THE LC (AND EC), LEADS TO THE PATHOLOGICAL HALLMARKS OF AD INCLUDING HIPPOCAMPAL PATHOLOGY. DEMONSTRATING HYPERACTIVATION AND PROTEOSTASIS DEFICITS IN THE LC AS INSTIGATORS OF HIPPOCAMPAL PATHOLOGY, PARTICULARLY, SELECTIVE NEURONAL LOSS PROVIDES MECHANISTIC INSIGHT AS TO WHY THESE ARE KEY NEURAL NETWORK CHANGES IN DISEASE. TO MODEL ALZHEIMER’S PATHOLOGY, WE FOCUS ON THE LC AND HPC TO IDENTIFY HOW THESE REGIONS ARE DISRUPTED WHEN PROTEOSTASIS SLOWS DOWN AND HOW HYPEREXCITATION IMPACTS THESE FUNCTIONS. WE WILL TRACK EARLY ELECTROPHYSIOLOGICAL CHANGES IN THE LC AND HPC WHEN PROTEINS LIKE BETA-AMYLOID (A) AND TAU START ACCUMULATING AND ASSESS HYPERAROUSAL/EXCITATION USING ELECTROPHYSIOLOGICAL MEASUREMENTS. OVER TIME, HYPEREXCITATION REDUCES CLEARANCE OF ABERRANT PROTEINS RESULTING IN A POSITIVE FEEDBACK LOOP OF PROTEOSTASIS LOSS AND HYPERAROUSAL, AND CASCADE TO HIPPOCAMPUS AND MEMORY LOSS. WE IDENTIFY THE TYPE OF NEURONS THAT ARE MOST VULNERABLE TO HYPEREXCITATION/AROUSAL-PROTEOSTASIS CHANGES IN THIS NETWORK, DESTABILIZING EXCITATORY-INHIBITORY HOMEOSTASIS. FINALLY, WE TEST IF DAMPENING HYPEREXCITATION OR PROTEOSTASIS RESTORATION IMPROVES COGNITIVE FUNCTION AND REVERSES PATHOLOGICAL CHANGES IN OUR MODEL. OUR GOAL IS TO USE OBSERVATIONS FROM ALL THE PARADIGMS TO IDENTIFY IF THESE BIOLOGICAL CHANGES AND PATHOLOGICAL SPREAD OF DISEASE CAN BE ANALYZED USING COMPUTATIONAL TOOLS TO PREDICT THE PATTERNS AND EVENTS LEADING TO AD AND TO TEST IF WE CAN USE AS A DISEASE RISK SCORE.

USING TRANSCRIPTION FACTORS TO ENHANCE TRANSPLANTED CELL SURVIVAL FOR SCI REPAIR

TARGETING STROKE-INDUCED BRAIN SWELLING IN OBESE SUBJECTS: ROLE OF VEGF

IMPACT OF SENSORY IMPAIRMENTS ON MOVEMENT IN CHILDREN WITH CEREBRAL PALSY

IMPACT OF BDNF SNP ON STROKE-INDUCED PLASTICITY AND MOTOR FUNCTION

ALLELIC CHOICE IN RETT SYNDROME

HDAC6: A TARGET FOR REGENERATION FOLLOWING INJURY IN THE NERVOUS SYSTEM

A NOVEL COMBINATORIAL APPROACH TO RESTORE MOTOR FUNCTION AFTER SPINAL CORD INJURY

MOTOR CORTEX ELECTRICAL STIMULATION TO AUGMENT SPONTANEOUS RECOVERY AFTER CHRONIC SUBCORTICAL STROKE

BENFOTIAMINE IN ALZHEIMER'S DISEASE: A PILOT STUDY

MOLECULAR REGULATION OVER THE DECLINE IN LONG-DISTANCE CORTICOSPINAL AXON REGENERATIVE ABILITY DURING DEVELOPMENT - CORTICOSPINAL NEURONS (CSN) RESIDE IN THE NEOCORTEX, AND EXTEND AXONS TO SPECIFIC SEGMENTAL TARGETS IN THE SPINAL CORD FORMING THE CORTICOSPINAL TRACT (CST). CSN CRITICALLY CONTROL VOLUNTARY MOVEMENT AND ARE CENTRALLY INVOLVED IN RECOVERY FROM PARALYSIS ORIGINATING FROM MULTIPLE CAUSES, E.G., STROKE, SPINAL CORD INJURY (SCI), CEREBRAL PALSY, ETC.. CSN DEGENERATION IN AMYOTROPHIC LATERAL SCLEROSIS (ALS), ALONG WITH DEGENERATION OF SPINAL MOTOR NEURONS, CAUSES SPASTICITY AND PARALYSIS. RECOVERY IN ALL THESE DISTINCT CAUSES OF PARALYSIS WOULD ULTIMATELY REQUIRE LONG-DISTANCE CST REGENERATION, WHICH REMAINS AN UNATTAINED GOAL IN REGENERATIVE NEUROSCIENCE. PREVIOUS WORK USING NEONATAL LESIONS HAS ESTABLISHED CST REGENERATIVE ABILITY DECLINES FROM DEVELOPMENT INTO ADULTHOOD. WHEN THE CST IS DAMAGED IN EARLY LIFE, THERE IS GREATER PLASTICITY AND REGENERATION AS COMPARED TO SIMILAR LESIONS IN THE ADULT. HOWEVER, DESPITE THIS WORK, WE STILL DO NOT KNOW WHEN LONG-DISTANCE REGENERATIVE ABILITY IS LOST DURING DEVELOPMENT. THIS IS BECAUSE A KEY LIMITATION OF THESE ESTABLISHED NEONATAL LESION MODELS IS THAT THEY MASSIVELY DISRUPT THE SPINAL ENVIRONMENT AND THEREBY INTERFERE WITH THE NORMAL PROCESS OF LONG- DISTANCE CSN AXON EXTENSION DURING DEVELOPMENT. THIS LIMITS THE ABILITY OF THESE LESION PARADIGMS TO ASSESS THE ABILITY OF THE CNS TO SUPPORT LONG-DISTANCE REGENERATION. WE RECENTLY ESTABLISHED A NOVEL MICROSURGICAL APPROACH TO AXOTOMIZE THE CST DURING DEVELOPMENT, WHILE LEAVING THE SPINAL ENVIRONMENT RELATIVELY INTACT. WE IDENTIFIED THAT LONG-DISTANCE CST REGENERATIVE ABILITY IS LOST AT DIFFERENT TIMES AT DISTINCT SPINAL LEVELS– AT POSTNATAL DAY 4 (P4) THE CST CAN FULLY REGENERATE WHEN LESIONED AT THORACIC T11, BUT FAILS TO DO SO WHEN LESIONED AT CERVICAL C2. OUR RESULTS INDICATE THAT THE LOSS OF LONG-DISTANCE CST REGENERATIVE ABILITY CLOSELY PARALLELS THE DEVELOPMENTAL TIMELINE OF NORMAL CST GROWTH INTO THE SPINAL CORD, WHICH SUGGESTS THAT THE NORMAL PROCESS OF DIFFERENTIATION, BOTH IN CSN AND IN THE SPINAL CORD, RESULTS IN THE LOSS OF LONG-DISTANCE REGENERATIVE ABILITY. THIS PROPOSAL INVESTIGATES THE HYPOTHESIS THAT INHIBITION OF DIFFERENTIATION TO PROLONG THE IMMATURE DEVELOPMENTAL STATE WILL EXTEND THE TIME WINDOW WHEN LONG-DISTANCE REGENERATION IS POSSIBLE. SPECIFICALLY, WE WILL MANIPULATE THE FUNCTION OF RE1 SILENCING TRANSCRIPTION FACTOR (REST), A GLOBAL REPRESSOR OF NEURAL DIFFERENTIATION, TO TEST THIS HYPOTHESIS. BUILDING ON THIS FOUNDATION, WE WILL FIRST MANIPULATE REST FUNCTION (BOTH GAIN- AND LOSS-OF FUNCTION) AT DISTINCT SPINAL LEVELS TO INVESTIGATE WHETHER THIS AFFECTS THE ABILITY OF THESE SPINAL SEGMENTS TO SUPPORT LONG-DISTANCE CST REGENERATION (AIM1). WE WILL MANIPULATE REST FUNCTION IN CSN TO SIMILARLY INVESTIGATE LONG-DISTANCE CST REGENERATION (AIM 2). FINALLY, WE WILL USE SINGLE CELL PROFILING TO INVESTIGATE POTENTIALLY DISTINCT MOLECULAR EFFECTS OF MICROLESIONS AT DISTINCT SPINAL LEVELS ON DISTINCT CSN SUBSETS DEPENDING ON THEIR DEVELOPMENTAL STATE (AIM 3). TOGETHER, OUR WORK WILL DISCERN NOVEL MOLECULAR MECHANISMS OF HOW LONG-DISTANCE CST REGENERATION IS REGULATED DURING DEVELOPMENT, THEREBY PROVIDING A MECHANISTIC FRAMEWORK FOR SUBSEQUENT IDENTIFICATION OF MOLECULES THAT CAN BE USED TO EFFECT LONG-DISTANCE CST REGENERATION IN THE ADULT CNS.

THE ROLE OF CORTICOSPINAL NEURONS IN THE RECOVERY OF DEXTEROUS FORELIMB FUNCTION AFTER SPINAL CORD INJURY - PROJECT SUMMARY: THE ADVERSE EFFECTS OF SPINAL CORD INJURY (SCI) ON CORTICOSPINAL FUNCTION ARE NOT RESTRICTED TO THE DAMAGED SPINAL CORD, BUT ALSO DISRUPT MOTOR REPRESENTATIONS WITHIN THE CORTEX. SCI RESULTS IN ALTERED CORTICAL MAPS THAT REPRESENT MOTOR OUTPUT, WITH REPRESENTATIONS ABOVE THE LEVEL OF INJURY EXPANDING INTO AFFECTED CORTICAL AREAS. REHABILITATION IS NECESSARY FOR BOTH THE RECOVERY OF CORTICOSPINAL-DEPENDENT FORELIMB FUNCTION AND THE COMMENSURATE REORGANIZATION OF DISRUPTED CORTICAL MOTOR MAPS. BOTH THE UNDERLYING CIRCUIT MECHANISMS THAT SUPPORT CORTICAL REORGANIZATION AFTER SCI AS WELL AS THE NECESSITY FOR THE REORGANIZED CIRCUITRY TO SUPPORT FUNCTIONAL RECOVERY, REMAIN UNKNOWN. FOR INJURED CORTICOSPINAL NEURONS TO CONTRIBUTE TO FUNCTIONAL RECOVERY, THEY MUST BE INTEGRATED INTO CORTICAL MOTOR NETWORKS. THE LONG-TERM GOAL IS TO DEVELOP THERAPEUTIC INTERVENTIONS FOR SUPPORTING FUNCTIONAL RECOVERY AFTER SCL THE OVERALL OBIECTIVE FOR THIS PROPOSAL IS TO DETERMINE HOW SPECIFIC REHABILITATION AFTER SCI PROMOTES REMODELING OF CORTICOSPINAL CIRCUITS AND THE CONTRIBUTION OF INJURED CORTICOSPINAL NEURONS TO MOTOR RECOVERY. THE CENTRAL HYPOTHESIS IS THAT CORTICOSPINAL-DEPENDENT REHABILITATION AFTER SCI DIRECTS THE STRUCTURAL REMODELING OF INJURED CORTICOSPINAL NEURONS RESULTING IN THEIR INCORPORATION INTO FUNCTIONAL MOTOR ENSEMBLES. THE RATIONALE FOR THE PROPOSED RESEARCH IS THAT DETERMINING THE PROPERTIES OF REHABILITATIVE TRAINING THAT PROMOTE SUCCESSFUL CORTICOSPINAL CIRCUIT INCORPORATION INTO CORTICAL MOTOR NETWORKS AFTER SCI WILL BE CRUCIAL FOR DEVELOPING EFFECTIVE REHABILITATIVE STRATEGIES. THE FOLLOWING THREE SPECIFIC AIMS ARE PROPOSED: 1) IDENTIFY THE NATURE OF STRUCTURAL AND CONNECTIVITY CHANGES THAT OCCUR IN INJURED CORTICOSPINAL NEURONS DURING REHABILITATION-MEDIATED RECOVERY FROM SCI; 2) IDENTIFY THE CHANGES IN THE FUNCTIONAL CONNECTIVITY OF INJURED CORTICOSPINAL NEURONS DURING REHABILITATION-MEDIATED RECOVERY FROM SCI; AND 3) IDENTIFY THE CONTRIBUTION OF INJURED CORTICOSPINAL NEURONS TO MOTOR RECOVERY AFTER SCL FOR THE FIRST AIM, THE APPROACH WILL BE TO IMAGE STRUCTURAL CHANGES OF INJURED CORTICOSPINAL DENDRITIC ARBORS IN RESPONSE TO REHABILITATION. IN THE SECOND AIM, THE APPROACHES WILL BE TO USE 2-PHOTON IMAGING TO RECORD THE ACTIVITY OF INJURED CORTICOSPINAL NEURONS DURING REHABILITATION AND TO USE RETROGRADE TRANSSYNAPTIC TRACING TO IDENTIFY PRESYNAPTIC INPUTS. IN THE THIRD AIM, THE APPROACH WILL BE TO OPTOGENETICALLY CONTROL INJURED CORTICOSPINAL NEURONS IN AWAKE, BEHAVING MICE TO DETERMINE THEIR CONTRIBUTION TO RECOVERY. THE PROPOSED STUDIES ARE INNOVATIVE IN THAT THEY SHIFT THE FOCUS OF SPINAL CORD REHABILITATION ONTO THE CIRCUIT MECHANISMS OF CORTICAL NETWORK PLASTICITY. THE PROPOSED STUDIES ARE SIGNIFICANT BECAUSE THEY WILL ELUCIDATE THE MECHANISMS BY WHICH CIRCUIT REMODELING INFLUENCES RECOVERY AND WILL INFORM COMBINATORIAL STRATEGIES THAT TARGET CORTICAL PLASTICITY TO FULLY REALIZE THE EFFECTS OF AXONAL SPROUTING AND REGENERATION. THE EXPECTATION IS THAT COMPLETION OF THE PROPOSED RESEARCH WILL DETERMINE THE ROLE FOR INJURED CORTICOSPINAL NEURONS IN THE RECOVERY OF FUNCTIONAL MOTOR NETWORKS AFTER SCI_ THESE FINDINGS WILL ESTABLISH A FOUNDATION TO GUIDE THE DEVELOPMENT OF THERAPEUTIC STRATEGIES TARGETING MOVEMENT RECOVERY AFTER CNS INJURY.

INJURY AND ADAPTATION IN THE DEVELOPING RAT CORTICOSPINAL AND RUBROSPINAL TRACTS

AXONAL TRANSPORT AND LOCAL TRANSLATION IN NEUROPATHIC PAIN

CROSS-TALK BETWEEN SARMOPTOSIS AND FERROPTOSIS IN HEMORRHAGIC STROKE - PROJECT SUMMARY/ABSTRACT BRAIN BLEEDING IN THE PARENCHYMA, ALSO KNOWN AS INTRACEREBRAL HEMORRHAGE (ICH), REPRESENTS 10 TO 30 PERCENT OF STROKES IN DISTINCT COUNTRIES AROUND THE WORLD. DESPITE ITS LOWER INCIDENCE THAN ISCHEMIC STROKE, ICH CARRIES HIGHER RATES OF MORTALITY AND MORBIDITY. WORSE OUTCOMES IN HUMANS WITH ICH HAVE BEEN CORRELATED WITH DAMAGE TO AXONS IN AREAS OF THE BRAIN LIKE THE THALAMUS OR THE STRIATUM WHERE BLEEDING FROM SMALL, PENETRATING ARTERIES (OFTEN AFFECTED BY LONGSTANDING HYPERTENSION) TENDS TO OCCUR. RED BLOOD CELLS ACCUMULATE IN ICH AND LYSE AND RELEASE TOXINS. THESE INCLUDE HEMOGLOBIN (HGB) AND ITS BREAKDOWN PRODUCT HEMIN . ADDITIONAL UNDERSTANDING OF HOW HGB/HEMIN LEADS AXONS TO DEGENERATE IN ICH IS A STRATEGY TO FULLY ASSESS AND OVERCOME DISABILITY FROM THIS CONDITION. EXCITINGLY, A MOLECULAR PATH FOR PROGRAMMED AXONAL DESTRUCTION HAS BEEN DEFINED INVOLVING THE PROTEIN, STERILE ALPHA AND TOLL/INTERLEUKIN RECEPTOR-1 (TIR) MOTIF CONTAINING 1 (SARM1). SARM1'S DEGENERATIVE ACTIVITIES IN THE AXON EMERGE FROM ITS NADASE (NAD+ DEGRADING) ACTIVITY IN THE TIR DOMAIN. THIS DOMAIN'S IMPORTANCE IS HIGHLIGHTED BY ITS CONSERVATION FROM PLANTS TO HUMANS. IN PRELIMINARY STUDIES, WE FOUND THAT GERMLINE DELETION OF FULL LENGTH SARM1 PREVENTS AXONAL DAMAGE IN THE STRIATUM FOLLOWING ICH. SINCE PREVIOUS STUDIES FROM OUR LAB HAVE SHOWN THAT HEMIN INDUCES FERROPTOSIS, AN IRON-DEPENDENT, CASPASE-INDEPENDENT FORM OF PROGRAMMED NECROSIS, FOLLOWING EXPERIMENTAL ICH, WE ARE SEEKING TO UNDERSTAND THE CROSS TALK BETWEEN FERROPTOSIS AND SARM1-MEDIATED AXONAL DESTRUCTION (SARMOPTOSIS). OF NOTE, WE FOUND THAT CORTICAL NEURONS CULTURED FROM THE SARM1 KNOCKOUT MICE ARE PARTIALLY RESISTANT TO FERROPTOTIC STIMULI (HEMIN AND ERASTIN). MOREOVER, AGENTS SUCH AS P7C3 THAT AUGMENT THE CELL'S ABILITY TO MAKE NAD+ PROTECT AGAINST FERROPTOSIS; CONVERSELY, FK866, WHICH IMPEDES THE SAME NAD+ SYNTHESIZING PATHWAY MAKES FERROPTOSIS WORSE. FROM THESE PRELIMINARY STUDIES WE PROPOSE THREE AIMS DESIGNED TO UNDERSTAND THE INTERPLAY BETWEEN FERROPTOSIS AND SARMOPTOSIS IN ICH. IN THE FIRST AIM, WE WILL LEVERAGE MICE THAT POSSESS A GERMLINE, HOMOZYGOUS, SINGLE AMINO ACID MUTATION (E642A) IN THE NADASE DOMAIN OF SARM1. LIKE THE SARM1-/- MICE, THE NEURONS OF THESE MICE ARE RESISTANT TO AXOTOMY IN VITRO AND IN VIVO. THEY WILL BE USED TO PROBE WHETHER THE NADASE ACTIVITY IS CRITICAL FOR FERROPTOSIS IN VITRO AND ICH IN VIVO. IN THE SECOND AIM, WE WILL ADDRESS THE HYPOTHESIS THAT HEMIN-INDUCES AXONAL DEGENERATION VIA LOCAL FERROPTOTIC PATHWAYS THAT TRIGGER SARM1 ACTIVATION. WE WILL USE SENSORY NEURONS CULTURED IN CAMPENOT CHAMBERS TO ISOLATE CELL BODIES FROM AXONS AND QUERY THEIR POSSIBLY DISTINCT RESPONSES TO LETHAL CONCENTRATIONS OF HEMIN. WE WILL ALSO PROBE THE ROLE OF FERROPTOSIS AND SARMOPTOSIS IN THE DESTRUCTION OF CORTICOSPINAL TRACT AXONS IN VIVO. IN THE LAST AIM, WE WILL SEEK TO UNDERSTAND WHETHER INTERVENTIONS THAT TARGET FERROPTOSIS BY ACTING IN THE NUCLEUS (CELL BODY) CAN BE COMBINED WITH SARM1 DELETION (THAT ACTS PRIMARILY IN AXONS) IN MICE TO REDUCE DAMAGE AND IMPROVE BEHAVIORAL RECOVERY OVER EACH AGENT TESTED ALONE. AT THE CULMINATION OF THESE STUDIES, NEW MECHANISTIC INFORMATION WILL EMERGE REGARDING AXONAL AND CELL BODY LOSS FOLLOWING ICH WITH CLEAR THERAPEUTIC IMPLICATIONS.

AUTOMATED ASSESSMENT OF VISUOMOTOR FUNCTION IN CHILDREN WITH BRAIN INJURY

ADVANCED, MULTI-FUNCTION, INVERTED CONFOCAL MICROSCOPE FOR BNI STRUCTURAL AND FUNCTIONAL IMAGING CORE

NON-INVASIVE STIMULATION FOR IMPROVING MOTOR FUNCTION IN SPINAL CORD INJURY

ADVANCED MATERIALS FOR SAFE AND EFFECTIVE STIMULATION OF THE RAT CERVICAL SPINAL CORD

ROLE OF SPARC IN WOUND HEALING AFTER SPINAL CORD INJURY - PROJECT SUMMARY: SPINAL CORD INJURY (SCI) TRIGGERS A CELLULAR AND EXTRACELLULAR MATRIX (ECM) REMODELING RESPONSE TO CLOSE THE WOUND. THIS RESPONSE IS DRIVEN LARGELY BY ASTROCYTES FORMING A PROTECTIVE BORDER AROUND INFILTRATING CELLS. THIS PROCESS IS INCOMPLETE IN MOST MAMMALS, FAILING TO RESTORE GROWTH PROMOTING SUBSTRATES, AND LEADING TO EXCESSIVE FIBROSIS. FIBROSIS AFTER SCI IS CHARACTERIZED BY THE SECRETION AND DEPOSITION OF ECM COMPONENTS (EG. COLLAGEN) AND THE ESTABLISHMENT OF A CHRONIC SCAR, IN WHICH CONNECTIVE TISSUE REPLACES NORMAL PARENCHYMA. CONSERVED ROLES FOR NON-NEURAL WOUND HEALING MECHANISMS HAVE BEEN IDENTIFIED IN THE ASTROCYTE RESPONSE TO SCI, BUT THE MOLECULAR CUES THAT DRIVE FIBROSIS AFTER SCI ARE NOT FULLY CHARACTERIZED AND REPRESENT A CRITICAL GAP IMPEDING THE DEVELOPMENT OF STRATEGIES TO IMPROVE SPINAL CORD WOUND HEALING. THE LONG-TERM GOAL IS TO DEVELOP THERAPIES THAT PROMOTE WOUND CLOSURE AND LIMIT FIBROTIC PROCESSES THAT LEAD TO PERSISTENT SCARRING. SPARC (SECRETED PROTEIN ACIDIC AND CYSTEINE RICH) IS A CENTRAL MODULATOR OF BOTH WOUND HEALING AND FIBROSIS. THERE IS A ROBUST AND PERSISTENT INCREASE IN SPARC PROTEIN EXPRESSION IN BORDER-ASSOCIATED ASTROCYTES AFTER SCI AND CONSTITUTIVE KNOCKOUT OF SPARC IMPAIRS WOUND HEALING AND IMPACTS BOTH CELLULAR AND EXTRACELLULAR REMODELING. THE OVERALL OBJECTIVE OF THIS PROPOSAL IS TO DETERMINE THE ROLE OF ASTROCYTIC SPARC SIGNALING IN THE STRUCTURAL AND FUNCTIONAL RESPONSES TO SCI. THE CENTRAL HYPOTHESIS IS THAT ASTROCYTIC SPARC HAS AN ACUTE, EARLY, PRO-WOUND HEALING ROLE AND A SUB-ACUTE, LATE, PRO-FIBROTIC ROLE FOLLOWING SCI. THE FOLLOWING TWO SPECIFIC AIMS ARE PROPOSED: 1) CHARACTERIZE THE TEMPORAL ROLES OF ASTROCYTIC SPARC IN WOUND REMODELING FOLLOWING SCI; 2) DETERMINE THE EFFECT OF ACUTE AND SUB-ACUTE SPARC DELETION IN ASTROCYTES ON FUNCTIONAL RECOVERY AFTER SCI. THE APPROACH IN AIM 1 WILL BE TO TARGET ACUTE AND SUB-ACUTE SPARC EXPRESSION IN ASTROCYTES USING TEMPORAL CONTROL IN A CONDITIONAL KNOCKOUT MOUSE MODEL TO EVALUATE CELLULAR RESPONSES AND ECM COMPOSITION FOLLOWING CONTUSIVE SCI. IN AIM 2, THE APPROACH WILL BE TO ASSESS SENSORY AND MOTOR RECOVERY IN CONTROL AND ASTROCYTE-SPECIFIC CONDITIONAL KNOCKOUT MICE WITH ACUTE OR SUB-ACUTE SPARC DELETION FOLLOWING CONTUSIVE SCI. THE PROPOSED RESEARCH IS INNOVATIVE IN THAT IT CENTERS ON DEFINING THE TEMPORAL AND CELL-TYPE SPECIFICITY FOR A KEY DRIVER OF EPITHELIAL WOUND HEALING AND FIBROSIS IN THE CONTEXT OF SCI, CONNECTING THE FIELDS OF NON-NEURAL AND NEURAL WOUND HEALING. THE PROPOSED STUDIES ARE SIGNIFICANT AS AN UNDERSTANDING OF CONSERVED ROLES FOR NON- NEURAL WOUND HEALING PATHWAYS AFTER SCI, MAY LEAD TO THE DEVELOPMENT OF NEW THERAPEUTIC APPROACHES TO ALTER SCI PROGRESSION, ENHANCE REMODELING, AND LIMIT PATHOLOGICAL SCARRING. THE EXPECTATION IS THAT COMPLETION OF THE PROPOSED STUDIES WILL DETERMINE THE ROLE OF ASTROCYTE-DERIVED SPARC IN WOUND HEALING AND FIBROSIS AFTER SCI. THE IDENTIFICATION OF NEW REGULATORS OF WOUND HEALING AND FIBROSIS WILL ALLOW FOR THE SUBSEQUENT DEVELOPMENT OF THERAPIES TO INCREASE REPAIR BY REDUCING THE GROWTH-RESTRICTIVE ENVIRONMENT AFTER SCI.

USING NATURAL MOUSE MOVEMENT TO ESTABLISH A DEVELOPMENTAL "BIOMARKER" FOR CORTICOSPINAL DAMAGE - THE CORTICOSPINAL TRACT (CST) IS A CRITICAL CIRCUIT UNDERLYING SKILLED VOLUNTARY MOVEMENTS. DAMAGE TO THIS CIRCUIT DURING DEVELOPMENT CAN CAUSE PERMANENT, LONG-TERM MOVEMENT DISABILITY IN HUMANS. RECOGNIZING AND TREATING SUCH DEVELOPMENTAL CST DAMAGE IS CHALLENGING, LARGELY BECAUSE IMMEDIATELY AFTER THE LESION, THERE ARE LIMITED OR ALMOST NO FUNCTIONAL DEFICITS. HOWEVER, EARLY RECOGNITION AND INTERVENTION WITH THE APPROPRIATE TREATMENT MEASURES IS KEY TO REDUCING LONG-TERM DISABILITY. THE USE OF PRECLINICAL MOUSE MODELS, WHICH HAVE OTHERWISE PROVEN TO BE HIGHLY USEFUL IN FUNCTIONAL INVESTIGATIONS OF NERVOUS SYSTEM DEVELOPMENT, HAS BEEN LIMITED IN THIS REGARD. SINCE THE CST IS KNOWN TO CONTROL SKILLED MOVEMENTS, ESTABLISHED BEHAVIORAL TESTS IN MICE THAT INVESTIGATE CST FUNCTION REQUIRE TRAINING MICE IN SKILLED TASKS. THIS PRECLUDES THEIR APPLICATION IN NEONATAL MICE. FURTHER, MOUSE MODELS USED TO INVESTIGATE DEVELOPMENTAL CST DAMAGE, E.G. NEONATAL HYPOXIA OR SPINAL INJURIES, DO NOT ONLY DAMAGE THE CST; RATHER THEY DISRUPT MULTIPLE NEURAL PATHWAYS. IT THEREFORE REMAINS COMPLETELY UNKNOWN WHETHER THE CST CONTRIBUTES ONLY TO SKILLED MOVEMENTS IN ADULT MICE, OR WHETHER IT ALSO CONTRIBUTES TO THE DEVELOPMENT OF NATURAL, INNATE MOTOR ABILITY DURING DEVELOPMENT, BEGINNING IN NEONATAL MICE. THIS LATTER POSSIBILITY WOULD SUGGEST THAT THERE ARE EARLY, ALBEIT SUBTLE, BEHAVIORAL CORRELATES OF DEVELOPMENTAL CST INJURY IN MICE. WE RECENTLY DEVELOPED A NEW MICROSURGICAL APPROACH TO SPECIFICALLY DISRUPT THE DEVELOPING CST IN NEONATAL MICE. IN ADDITION, WE HAVE ALSO ESTABLISHED THE USE OF MOTION SEQUENCING (MOSEQ), A NEW MACHINE LEARNING AND ARTIFICIAL INTELLIGENCE PLATFORM, TO LONGITUDINALLY INVESTIGATE THE DEVELOPMENT OF NATURAL MOVEMENTS IN NEONATAL MICE. OUR PRELIMINARY RESULTS USING MOSEQ SUGGEST THAT DEVELOPMENTAL DAMAGE TO THE CST RESULTS IN SPECIFIC CHANGES TO MOVEMENT STRUCTURES IN MICE, AS EARLY AS P12; THESE EXTEND INTO MATURITY AT P35. FURTHER, ANALYSIS OF THESE P35 MICE USING CONVENTIONAL METRICS OF LOCOMOTION SUCH AS THE CATWALK, DID NOT IDENTIFY ANY DEFICITS, HIGHLIGHTING THE SENSITIVITY OF MOSEQ IN IDENTIFYING CHANGES IN MOUSE MOVEMENTS. THIS PROPOSAL INVESTIGATES THE HYPOTHESIS THAT THE CST CONTROLS DEVELOPMENT OF NATURAL MOUSE MOVEMENTS, AND NOT ONLY SKILLED MOVEMENTS AT MATURITY. WE WILL USE MOSEQ TO ANALYZE FEZF2 KNOCK OUT (FEZF2 KO) MICE IN WHICH THE CST IS NEVER ESTABLISHED DURING DEVELOPMENT (AIM1), AS WELL AS MICE THAT UNDERGO MICROSURGICAL LESIONS TO DISRUPT SPINAL CONNECTIVITY OF THE CST AT DISTINCT DEVELOPMENTAL TIMES (AIM2). TOGETHER, OUR WORK WILL IDENTIFY NOVEL FUNCTIONAL READOUTS OF DEVELOPMENTAL DAMAGE TO THE CST USING NATURAL MOUSE MOVEMENTS. THIS NEW UNBIASED QUANTITATIVE APPROACH TOWARD INVESTIGATING THE EARLIEST BEHAVIORAL SIGNS OF CORTICOSPINAL DYSFUNCTION IN MICE WHICH WILL HAVE EVENTUAL APPLICATION IN INVESTIGATIONS OF DESCENDING CIRCUITS OF MOTOR CONTROL, AS WELL AS MULTIPLE PRECLINICAL MODELS OF DEVELOPMENTAL DAMAGE SUCH AS NEONATAL HYPOXIA OR SPINAL INJURIES.

LOSS OF A COHORT OF AXONAL MRNAS AS A RESULT OF REDUCED SMN LEVELS.

TRANSCRANIAL DIRECT CURRENT STIMULATION AND MOTOR TRAINING IN CHRONIC STROKE

AN ADVANCED, HIGH-THROUGHPUT IMAGING SYSTEM FOR THE BNI STRUCTURAL AND FUNCTIONAL IMAGING CORE - THIS APPLICATION IS A REQUEST FOR FUNDS TO ACQUIRE A ZEISS AXIOSCAN 7 SLIDE SCANNER FOR THE STRUCTURAL AND FUNCTIONAL IMAGING CORE AT THE BURKE NEUROLOGICAL INSTITUTE. THE INSTITUTE, DEDICATED TO FINDING CURES FOR CHRONIC NEUROLOGICAL DISABILITIES, REQUIRES AN AUTOMATED IMAGE ACQUISITION SYSTEM TO ADVANCE THEIR NIH-FUNDED RESEARCH. THE MANDATE OF THE IMAGING CORE FACILITY IS TO PROVIDE ACCESS TO A RANGE OF LIGHT MICROSCOPY SYSTEMS AND PROVIDE TECHNICAL ASSISTANCE WITH IMAGE ACQUISITION, PROCESSING, AND ANALYSIS TO ALL RESEARCH GROUPS WITHIN BNI. WHILE THE INSTITUTE HAS INVESTED IN HIGH-RESOLUTION CONFOCAL AND MULTIPHOTON MICROSCOPY SYSTEMS, IT LACKS MODERN EPIFLUORESCENCE AND BRIGHTFIELD IMAGING SYSTEMS. EXISTING MICROSCOPY SOLUTIONS ARE AGING AND LACK CRITICAL FEATURES FOUND IN THE PROPOSED AXIOSCAN 7 SYSTEM. THIS SCARCITY OF RESOURCES POSES A SIGNIFICANT BOTTLENECK FOR THE DIVERSE NIH-FUNDED RESEARCH PROGRAMS AT THE INSTITUTE. THE AXIOSCAN 7 SLIDE SCANNER, WITH ITS AUTOMATED FULL SLIDE WORKFLOW, UNLIMITED Z-STACK ACQUISITION PARAMETERS, AND ADVANCED DECONVOLUTION ALGORITHMS, OFFERS AN OPTIMAL COMBINATION OF FEATURES FOR LARGE AREA TISSUE SCANNING AND STITCHING. THE PROPOSED SYSTEM WILL GREATLY EXPEDITE RESEARCH PROGRESS BY REDUCING ACTIVE USER TIME DURING IMAGE ACQUISITION. FURTHERMORE, THE SURROUNDING AREA LACKS OTHER AUTOMATED, HIGH-THROUGHPUT MICROSCOPY RESOURCES ACCESSIBLE TO BNI INVESTIGATORS, MAKING THE AXIOSCAN 7 AN INDISPENSABLE ASSET. ITS IMPLEMENTATION WILL NOT ONLY BENEFIT CURRENTLY NIH-FUNDED RESEARCH PROGRAMS BUT ALSO CATALYZE THE PROGRESS OF STUDIES THAT ARE YET TO REACH THE MATURITY REQUIRED FOR NIH FUNDING. THE PROPOSED RESEARCH PROJECTS UNDERSCORE THE SHARED NEED FOR AN ADVANCED SLIDE SCANNING SYSTEM. IN CONCLUSION, THE EXTENSIVE CAPABILITIES AND EFFICIENT WORKFLOW OF THE AXIOSCAN 7 WILL EXERT A SUSTAINED, POWERFUL INFLUENCE ON THE CONDUCT OF NIH-FUNDED RESEARCH PROGRAMS AT THE BURKE NEUROLOGICAL INSTITUTE AND SUPPORT BREAKTHROUGHS IN THE UNDERSTANDING AND TREATMENT OF CHRONIC NEUROLOGICAL DISABILITIES.

DISSECTING MOTOR CORTEX MODULATION OF NOCICEPTION DURING CHRONIC PAIN - PROJECT SUMMARY (SEE INSTRUCTIONS): THE HEAVY BURDEN OF CHRONIC PAIN AND THE OPIOID EPIDEMIC HAS PROMPTED AN URGENT, WORLDWIDE SEARCH FOR ALTERNATIVE, NON-ADDICTIVE METHODS OF ANALGESIA. ONE PROMISING ALTERNATIVE IS NON-INVASIVE ELECTRICAL OR MAGNETIC STIMULATION OF THE MOTOR CORTEX (MC). WHILE MC STIMULATION (MCS) HAS REPEATEDLY BEEN FOUND TO REDUCE CHRONIC PAIN IN HUMAN SUBJECTS AND DECREASE NOCICEPTIVE BEHAVIORS IN RODENT MODELS, MAJOR QUESTIONS REMAIN ABOUT ITS MECHANISM OF ACTION AND HOW TO IMPROVE MCS EFFICACY. EVIDENCE SUGGESTS THAT MCS ANTINOCICEPTIVE EFFICACY INCREASES WHEN THE STIMULATION TARGETS THE REGION IN MOTOR CORTEX THAT CORRESPONDS TO THE BODY PART FROM WHICH PAIN ORIGINATES (SOMATOTOPICALLY MATCHING MCS) AND THAT MCS ANALGESIA REQUIRES ENDOGENOUS OPIOID ACTIVITY. HERE, I PROPOSE TO USE A RODENT MODEL OF TRIGEMINAL NEUROPATHIC PAIN TO ELUCIDATE THE UNDERLYING MECHANISM OF MCS ANTINOCICEPTION AND TO DEFINE KEY MCS FEATURES THAT PAIN CLINICIANS CAN USE TO IMPROVE MCS EFFICACY. TO CHARACTERIZE THE MCS MECHANISM, I WILL 1) QUANTIFY THE EFFICACY OF SOMATOTOPICALLY MATCHED MCS (SSMCS), 2) DETERMINE WHAT OPIOID RECEPTOR SUBTYPES ARE REQUIRED FOR MCS ANTINOCICEPTION, AND 3) IDENTIFY HOW ENDOGENOUS OPIOIDS MODULATE AN OPIOID-SENSITIVE MC DESCENDING CIRCUIT TO THE DESCENDING PAIN CONTROL PATHWAYS DURING SSMCS. TO DETERMINE THE EFFICACY OF MCS BETWEEN MATCHED AND OFF-TARGET MCS IN TWO DIFFERENT NERVE CONSTRICTION MODELS, I WILL USE CLASSIC BEHAVIORAL PARADIGMS ALONG WITH CUTTING-EDGE MACHINE LEARNING ALGORITHMS TO ANALYZE MOUSE BEHAVIORAL RESPONSES. I WILL THEN USE COMPLEMENTARY PHARMACOLOGICAL AND GENETIC MOUSE LINES TO DETERMINE HOW SSMCS IS IMPACTED BY DISTINCT OPIOID RECEPTOR TYPES. TO INTERROGATE THE COMBINED IMPACT OF MC SOMATOTOPY AND ENDOGENOUS OPIOID SIGNALING IN MCS ANALGESIA, I WILL FOCUS ON THE DESCENDING PROJECTION FROM MC TO DESCENDING PAIN CONTROL REGIONS, ROSTRAL VENTROMEDIAL MEDULLA (RVM) AND SPINAL TRIGEMINAL NUCLEUS CAUDALIS (SPVC). I WILL DETERMINE THE POSITIONS OF OPIOID RECEPTOR TYPES AND ENDOGENOUS OPIOID PEPTIDES ALONG THIS CIRCUIT AND THEN ASSESS THE IMPACT OF SSMCS WITH AND WITHOUT OPIOID SIGNALING ON NEURAL ACTIVITY IN MC, RVM, AND SPVC USING CUTTING-EDGE HIGH-DENSITY ELECTROPHYSIOLOGICAL TECHNIQUES. ALTOGETHER, THIS PROJECT WILL DETERMINE HOW MCS MODULATES NOCICEPTION THROUGH ENDOGENOUS OPIOID SIGNALING IN SOMATOTOPICALLY ALIGNED CIRCUITS. THE RESULTS WILL PROVIDE THE FIRST REPORT OF NEURAL ACTIVITY DURING AND AFTER SSMCS AT BOTH THE TARGET AND IN AN MC OUTPUT.

TRANSCRANIAL DIRECT CURRENT STIMULATION AND ROBOTIC TRAINING IN ADULTS WITH CEREBRAL PALSY

IMPACT OF MOTOR CONNECTIVITY ON EFFICACY OF HAND THERAPY IN CONGENITAL HEMIPLEGIA

THE KNOB SUPINATION TASK: A SENSITIVE TEST OF CORTICOSPINAL FUNCTION IN THE RAT

A SKILLED, AUTOMATED BEHAVIORAL TASK TO ASSESS MOTOR NETWORK FUNCTION DURING IN VIVO IMAGING IN MICE.

ELUCIDATING THE MECHANISMS OF NEUROPROTECTION OF HISTONE DEACETYLASE INHIBITION IN ISCHEMIC STROKE

CORTICOSPINAL NEURON PLASTICITY IN RESPONSE TO DEVELOPMENTAL SPINAL CORD AXOTOMY. - PROJECT ABSTRACT SPINAL CORD INJURY (SCI) IS A DEVASTATING CONDITION WITH LIMITED TREATMENT OPTIONS DUE TO THE RESTRICTED REGENERATIVE CAPACITY OF THE MATURE CENTRAL NERVOUS SYSTEM. DUE TO THE HETEROGENEITY OF SCI ALONG THE NEURAXIS, IT IMPAIRS VARIOUS CORTICOSPINAL CIRCUITRY THAT AFFECTS MOTOR CONTROL. THE CORTICOSPINAL TRACT (CST), ESSENTIAL FOR VOLUNTARY MOVEMENT, CONSISTS OF DIVERSE CORTICOSPINAL NEURON (CSN) SUBTYPES, YET THE ESTABLISHMENT OF THESE DISTINCT SPINAL-PROJECTING NEURONS REMAINS UNCLEAR. RECENT FINDINGS FROM THE SAHNI LAB HAVE IDENTIFIED EARLY MOLECULAR DIFFERENCES IN CSN SUBPOPULATIONS PROJECTING TO THE CERVICAL OR THORACOLUMBAR SPINAL CORD FROM CORTICAL REGIONS OUTSIDE THE CLASSICAL MOTOR CORTEX; HOWEVER, THIS MOLECULAR MAP IS INCOMPLETE. ADDITIONALLY, THE LAB HAS DEVELOPED A NOVEL MICROLESION TECHNIQUE TO AXOTOMIZE THE DEVELOPING CST, ENABLING THE STUDY OF CSN PLASTICITY AND CIRCUIT FORMATION DURING DEVELOPMENT. UNDERSTANDING HOW CSN PROJECTION TYPES AND THEIR POSTSYNAPTIC SPINAL CONNECTIVITY ARE ESTABLISHED IS CRUCIAL FOR DEVELOPING STRATEGIES FOR FUNCTIONAL RECOVERY AFTER SCI. THIS PROPOSAL AIMS TO INVESTIGATE THE MOLECULAR MECHANISMS UNDERLYING CSN DEVELOPMENT AND THEIR LONG-RANGE CORTICOSPINAL CIRCUITS. MY CENTRAL HYPOTHESIS IS THAT INTRINSIC REGULATORS GOVERN SEGMENTALLY DISTINCT CSN IDENTITIES AND SPINAL SYNAPTIC CIRCUITRY. SPECIFIC AIM 1 WILL IDENTIFY MOLECULAR REGULATORS OF CSN DIVERSITY AND DETERMINE WHETHER SEGMENTAL SPINAL TARGETING IS GOVERNED BY INTRINSIC MECHANISMS OR TARGET- DERIVED RETROGRADE SIGNALING. SPECIFIC AIM 2 WILL EXPLORE HOW ROUTE OF AXON EXTENSION INFLUENCES SYNAPTIC SPECIFICITY AND THEIR IMPLICATIONS FOR THE EVOLUTION OF CORTICAL-MOTONEURAL CONNECTIONS AND DEXTERITY. TO ACHIEVE THESE AIMS, WE WILL UTILIZE MOUSE MODELS AND EMPLOY TECHNIQUES SUCH AS VIRAL TRACING AND MICROLESION APPROACHES. THE FINDINGS WILL ENHANCE OUR UNDERSTANDING OF NEURODEVELOPMENTAL PROCESSES AND MAY LEAD TO NEW THERAPEUTIC STRATEGIES FOR SCI RECOVERY. WITH THE SUPPORT OF THIS F32 FELLOWSHIP, I WILL STRENGTHEN MY EXPERTISE IN VIRAL TRACING, MOLECULAR PROFILING, AND BIOINFORMATICS, PREPARING ME TO LEAD AN INDEPENDENT RESEARCH PROGRAM FOCUSED ON LEVERAGING DEVELOPMENTAL MECHANISMS TO REPAIR THE INJURED NERVOUS SYSTEM.