Rutgers Biomedical And Health Sciences

NIH Research Projects · FY 2025 · 2022-09

Most Alzheimer’s Disease (AD) patients experience severe motor impairment at the later stage of disease and 10 - 40% of AD patients exhibit signs of motor dysfunction at even earlier stages of AD. Furthermore, changes in motor function often precede other symptoms of AD as well as correlate with increased severity and mortality. Despite the frequent occurrence of motor dysfunction in AD patients, little is known about the mechanisms by which this behavior is altered. In several other neurological diseases, such as stroke and vascular parkinsonism (VP), cerebrovascular lesions underlie motor dysfunction during the progression of these diseases, especially in the basal ganglia. In addition, white matter lesions (WMLs), which are primarily considered a small vessel disease and characterized as focal abnormal myelination, are highly correlated with motor deficits in VP. Moreover, WMLs are strongly associated with the clinical risk of AD and may accelerate the clinical manifestation of the disease. Familial Danish Dementia (FDD) is another AD-like familial neurodegenerative disease associated with motor dysfunction, WML, and vascular impairment. However, it is unclear which pathogenic mechanisms produce vascular impairment, WML, motor dysfunction in AD and FDD. Since several clinical studies suggest a strong connection between vascular deficits in basal ganglia and motor dysfunction in several neurological diseases, we investigated these pathologic correlations in AD mouse model. We found a significant increase in fibrin deposits, demyelination, and axonal degeneration as well as a decrease in blood vessel density in the striatum of the aged AD mice which exhibited motor deficits. Furthermore, we found the depletion or destabilization of fibrin in AD mice improved their motor performance. Based on these findings, we hypothesize that fibrin deposits and vascular degeneration lead to Blood Brain Barrier (BBB) damage, aggravate inflammation and demyelination, as well as cause axonal degeneration, finally leading to motor dysfunction in AD and FDD. In this proposal we will analyze postmortem brain tissues of AD patients who clinically exhibited motor deficits in the early disease state and investigate the pathogenic mechanism of motor dysfunction in rodent models of AD and FDD using biochemical, histological, and genetic methods (expertise by MPI Ahn). We will also investigate how striatal fibrin deposits cause demyelination and motor dysfunction in AD by induction of resistant fibrin clots or depleting the coagulation factor FXIII. Furthermore, we will employ advanced Magnetic Resonance Imaging (MRI) techniques in a mouse model of AD, FXIII deficient AD mice and a knock-in rat model of FDD (expertise by MPI Dyke). Our techniques will interrogate the permeability of the BBB and assess cerebral blood flow, and WMLs seen in white matter hyperintensities as well as demyelination. Our long-term objective is to translate our findings in this proposal for the direct clinical MRI use in assessing permeability, demyelination and neurodegeneration in human subjects and developing therapeutics for AD and FDD.

NIH Research Projects · FY 2026 · 2022-09

Engaging in affiliative social interactions predicts health and longevity. The hormone oxytocin, which is released by specialized neurons in the hypothalamus, has been shown to play an important role in affiliative behaviors. However, the mechanisms that control activity of oxytocin neurons and by which these neurons might then control engagement in social interactions, have not yet been elucidated. Our long-term goal is to dissect the neuroendocrine mechanisms driving voluntary engagement in affiliative social interactions. Our findings will facilitate the design of therapeutic interventions aimed at addressing the widening loneliness pandemic, as well as ASD. The objective of this grant is to characterize the role of oxytocin neuron activity patterns in initiation and reinforcement of social interactions. The central hypothesis is that spiking activity in oxytocinergic neural circuits plays a critical role in spontaneous and stress-induced social engagement, and that activity in distinct oxytocinergic circuits specifically controls either initiation or reinforcement of social engagement. In our specific aims we will use automated behavioral analysis of video recordings of mice living together for days, behaviorally synchronized neuronal recordings, optogenetics and chemogenetics to determine if neuronal activity of identified oxytocin neurons predicts or tracks social interactions (AIM 1). We will use optogenetics and chemogenetics to manipulate neuronal activity of specific oxytocinergic circuits to determine if they are required for the initiation or reinforcement of affiliative interactions (AIM 2). We will test if stressful contexts increase activity in specific oxytocinergic neurons to promote engagement in affiliative social interactions, as a defensive mechanism (AIM 3). Our work will significantly contribute to the field, as it will establish mechanisms that can be therapeutically targeted to drive social engagement. The proposed research is innovative because we investigate activity patterns in neuroendocrine circuits that predict spontaneously initiated and reinforced social interactions, which has not been done before. Our results will lay the bases for novel behavioral interventions and medications that could be used to prevent the negative effects of social isolation.

NIH Research Projects · FY 2026 · 2022-09

PROJECT SUMMARY/ABSTRACT: This proposal is in application for a Pathway to Independence Award for Dr. Travis Lear at the Aging Institute at the University of Pittsburgh. Dr. Lear has extensive experience in the molecular biology of ubiquitination in aging, inflammation, autophagy, and lysosome biology. This K99/R00 would be a crucial step for Dr. Lear as part of his career goal of reaching research independence. The focus of Dr. Lear’s future lab will be to study the mechanisms of protein degradation and lysosomal activity in neurodegeneration in pursuit of uncovering new therapeutic avenues. Under this K99/R00 award, Dr. Lear would have protected time for additional training in neurobiology techniques and aging biology, and for career development to facilitate a successful transition to research independence. This mentorship team of experts in aging, lysosomal, and neurobiology combined with the scientific environment at the Aging Institute will be ideal for Dr. Lear’s training. The scientific expertise training will entail 1) in vitro primary tissue culture skill, 2) study of iPSC development, differentiation, and gene-editing, 3) generation and phenotypic characterization of mouse models of neurodegeneration. Dr. Lear’s leadership training will focus on 1) integration to scientific community through networking, 2) enhancing his mentoring skills, 3) improving grantsmanship. Successful completion of the proposed research and training plan will provide the knowledge and experience necessary to progress toward Dr. Lear’s career goal of becoming an independent investigator studying neuronal aging and Alzheimer’s Disease (AD). To accomplish this, Dr. Lear proposes to study a new mechanism of proteolytic control of lysosomal activation to augment processing of pathogenic tau protein aggregates in models of AD. Specifically, Dr. Lear has elaborated a model in which a key mTORC1 inhibitor protein, KPTN, is potently controlled by the E3 ubiquitin ligase PDZRN3, which ubiquitinates and fates KPTN for degradation. Also, unbiased screening yielded a small molecule KPTN activator which increases KPTN protein level, inhibits mTORC1 activity, and increases lysosomal number and activity. Excitingly, pharmacological augmentation of KPTN reduces tau protein aggregation in vitro, which has therapeutic implications for the treatment of Alzheimer’s Disease. The net effect of KPTN augmentation would therefore aid in clearance of toxic protein aggregates by activation of autophagy. This leads to the central hypothesis that PDZRN3 control of KPTN affects neuronal lysosomal activity and that genetic or pharmacological activation of KPTN may be an avenue to reduce tau protein aggregates. Two aims will interrogate this hypothesis: (1) To examine the mechanism and biologic effect of the PDZRN3-KPTN axis on tau-aggregation in vitro with inducible pluripotent stem cell (iPSC) and primary cell models, and (2) to examine this mechanism and effect using genetic and pharmacological approaches in animal models of tau spreading. Dr. Lear will also pursue a structured training plan including formal course work, conferences, and hands-on training of new techniques, under the guidance of primary mentor Dr. Toren Finkel, and co-mentors Dr. Stacey Rizzo and Dr. Bill Chen.

NIH Research Projects · FY 2025 · 2022-09

Project Summary The overall objective of this proposal is to elucidate specific crosstalk mechanisms between the integrated stress response (ISR) and circadian timekeeping, two fundamental biological processes in neurons. Circadian rhythm regulates neuronal differentiation, plasticity, and tissue regeneration and its disruption contributes to a variety of human health problems. Circadian clock genes are widely expressed in almost all cells. To function properly, the cellular clock must integrate and synchronize with cellular physiology and metabolism. ISR is a conserved intracellular signaling network for cells to respond to stressors and restore homeostasis. Little is known, however, on whether and how ISR integrates with the circadian clock, which forms a major gap in our understanding of homeostatic integration in neurons. Our recently published work indicates that ISR may be a conserved mechanism that couples cellular stress response to circadian timekeeping. Based on the published work and unpublished preliminary data, here we propose to test the overall hypothesis that ISR interacts with the mammalian circadian clock: ISR regulates fundamental clock properties including entrainment and circadian period, whereas the clock controls ISR response based on the time of day. We will leverage our expertise and unique mouse models to test the hypothesis using a combination of molecular, cellular, and behavioral approaches. Aim 1 will define a role for the ISR inhibitor IMPACT (imprinted and ancient gene protein) in regulating photic entrainment of the circadian clock. Aim 2 will identify a role for unfolded protein response and PERK (protein kinase R-like endoplasmic reticulum kinase) in circadian timekeeping. Aim 3 will elucidate eIF2 (eukaryotic translation initiation factor 2)-dependent translational control mechanisms in the circadian clock. The proposed work is innovative, because it utilizes new mouse genetic tools to address conceptually novel questions regarding the crosstalk mechanisms between ISR and the clock. The contributions are expected to be significant, because it is expected to uncover mechanistic links between the two fundamentally important cellular processes. Importantly, ISR frequently goes awry in complex brain disorders, which are often associated with disrupted daily rhythms in patients due to unknown mechanisms. As ISR can be targeted by FDA-approved drugs, understanding its role in circadian physiology may offer new opportunities to regulate the body clock function and to treat clock dysfunctions in these diseases.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The number of people impacted by Alzheimer’s Disease and Alzheimer’s Disease Related Dementias (ADRD) is 5 million in the US alone and is expected to grow, especially in Black communities. Black older adults have a 40-80% higher incidence of ADRD and approximately double the risk of underdiagnosis. Innovative use of survey-Medicare Claims linked data holds promise to advance our understanding of disparities in timely ADRD diagnosis and their contributing factors. However, no prior research has validated this approach. Also, prior research on risk factors for delayed ADRD diagnosis has focused on individual-level characteristics that are difficult to modify. I seek to fill this gap by validating the measure of timeliness of ADRD diagnosis with respect to the survival benefits and examining placed-based structural factors associated with disparities in timely ADRD diagnosis. Residential racial segregation and geographic features of health care access are important to consider together. While unequal distribution of health care resources in segregated areas may be the primary driver of the racial disparities in timely diagnosis, residential segregation may capture unobserved features of structural racism influencing health care utilization patterns by race. These factors can be mutually reinforcing drivers of poor diagnosis in the Black population. In the K99 phase, I will conduct a robust epidemiological analysis to examine the timeliness of ADRD diagnosis related to survival using the Health and Retirement Study and the National Health and Aging Trends Study. In the R00 phase, I will examine the association between residential racial segregation or health care access features and racial disparities in the timeliness of ADRD diagnosis while extending my inquiry to the cohort from the Reasons for Geographic and Racial Disparities in Stroke study. I will also examine how the magnitude of the association between health service characteristics and timely ADRD diagnosis differs by small-area characteristics (e.g., area deprivation) and individual characteristics (e.g., education). This research plan is complemented by a training plan that builds on my background in social work and sociology, and includes new training in (1) neurodegenerative diseases, clinical assessment, and diagnostic pathways; (2) application of algorithm-based dementia assessment in surveys; (3) utilization of Medicare claims data, and (4) gaining additional sociological knowledge in linking data in residential segregation, health care access, and racial disparities related to timely ADRD diagnosis. The combined research and training plans will prepare me for a successful independent research career focused on identifying modifiable determinants of Alzheimer’s disease and related dementias, and study of the effects of policies and interventions to reduce racial disparities in ADRD. Together, these have the potential to improve prevention and care in ADRD. My mentorship team is uniquely poised to assist me in achieving my training and research goals and ensure my successful transition to independent investigator status.

NIH Research Projects · FY 2025 · 2022-09

Due to the high stakes of healthcare, the primary barrier is the extremely low tolerance of errors in healthcare practice, which requires extremely high sensitivity and specificity of any modelling. However, nearly all Machine learning (ML) models focus on improving the accuracy. It cannot yet reach both extremely high sensitivity and specificity using healthcare data. Separate screening and confirmatory ML tools are proposed to achieve very high sensitivity and specificity. Moreover, many ML algorithms suffer from the lack of clear explanations, such as deep learning and neural networks, and would unlikely meet the FAIR criteria. Cancer is the second leading cause of death in the U.S. The number of cancer survivors continues to grow; unfortunately, so does the number of non-cancer deaths in cancer patients. However, nearly all `omic and large population studies focused on binary outcomes (cancer death or recurrence). Therefore, there is an urgent need to better understand and reduce non-cancer deaths in cancer patients, using `omic and population data. To address these problems, the project will develop screening and confirmatory ML to model cancer and noncancer deaths in breast, colorectal, prostate and lung cancer patients using `omic data and electronic health records (EHR). The proposed research will result in fundamental contribution to ML tools, workflows and methods to make novel use of `omic and EHR data for cancer care. It timely meets the urgent needs in precise reduction of non-cancer deaths. This project also uniquely addresses the Transformative Data Science research theme. The interdisciplinary collaboration in this project as outlined in the Collaboration Plan will offer a diverse basis for creative problem solving and validation. The proposal has 3 broader impacts: 1) The developed novel ML algorithms and technology will enable physicians to more precisely prognosticate and treat cancer patients based on their risk of multicategory deaths. 2) The research program will support and nurture undergraduate and graduate researchers. 3) The proposed research program will support high school and undergraduate students both in the conduct of research and in awareness of ML usefulness.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT Medications for opioid use disorder (MOUD) are a key tool in reducing harms of the opioid epidemic. Yet only a minority of those with OUD initiate treatment, early discontinuation is typical, and disparities are endemic. People with disabilities are at especially high risk and epitomize the challenges of OUD with multimorbidity. Preliminary analyses identified 45,035 fatal opioid overdoses among Medicare disability beneficiaries (MDBs) from 2008-2016, and continuing under-utilization and disparities in MOUD, including among overdose survivors. With its wide influence in the health care system, Medicare's role is vital; it is essential to examine the Medicare system's successes and failures in engaging and retaining MDBs in treatment. Several recent policy changes are promising, with important implications for other payers, but their impact across beneficiary subgroups, time and communities needs to be better understood to inform action to improve uptake and reduce disparities. This study, responding to RFA-DA-22-037, will use national Medicare data linked with the National Death Index, Medicaid claims, community resources, prescription drug plan (PDP) formulary policies, and other data sources to assess how policy, community, provider and patient factors interact to shape MOUD initiation and retention, and in turn overdose and other clinical outcomes. With annual updates through 2025, the project will provide a powerful framework for assessing evolving treatment patterns and outcomes in a rapidly evolving environment, as well as potential changes in policy impacts over time. We will assess the drivers of racial/ethnic and other disparities in access; MOUD changes following policy and formulary changes by Medicare and its PDPs; and how these policies interact with the evolving MOUD provider system, community resources and patient characteristics. We will analyze trends and disparities in MOUD treatment and overdoses among MDBs. In cohorts of beneficiaries with new OUD diagnoses or non-fatal overdoses, we will assess factors associated with treatment initiation and retention, and association of treatment with clinical outcomes including non-fatal and fatal overdose. We will assess MOUD uptake across community, provider, and patient subgroups; changes in MOUD treatment patterns associated with the shift to tele-health; and associated changes in the MOUD treatment network serving MDBs. We will examine the sequelae of changes in formulary policies across Medicare's more than 6000 PDPs, including prior authorization requirements for MOUD, across beneficiary subgroups. Expanded reimbursement for tele-MOUD and elimination of prior authorization have the potential to save many lives, but it is critical to better understand their impact on access and disparities. An active dissemination strategy supported by a Stakeholder Advisory Board, complementing peer-reviewed publication, will support translation into evidence-informed policy. Results of this innovative and comprehensive assessment of the multi-level factors shaping MOUD uptake and outcomes among MDBs will have important implications for policy and practice across patient subgroups, payers and health care systems.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Developing efficient cognitive training for cognitively intact older adults (OA) is a major public health goal, due to its potential for reducing age-related cognitive decline and Alzheimer’s disease/dementia risk. Executive Control (EC) is a relevant training target since it declines with aging and is critical for multi-tasking in daily life. EC training programs for OA have been shown to lead to cognitive improvements, but it remains unclear whether it enhances multi-tasking ability and transfer to different tasks and context, such as everyday life. Most EC training protocols fail to adopt ecological tasks, which limit its clinical relevance and generalization. A promising EC training approach is Emphasis Change (EmCh), which has shown to benefit both younger and OA. Similar approaches have shown training-transfer in OA, suggesting that EmCh may be an appropriate method to induce transfer in OA. To date, EmCh has not been applied in ecological tasks that simulate a daily life situation, nor has it been implemented remotely through a web-based interface. The web-based training is an advancement since it can be delivered at home and is easily scalable. Recently, remote interventions have been relevant to OA due to the COVID-19 pandemic, which limits in-person research participation. The current proposal seeks to bridge the above gaps by implementing EmCh through the Breakfast Game (B-Game), a web-based ecological training platform that simulates a daily life environment and is feasible in OA. The Aims 1 and 2 (K99 phase) of this proposal will be based on a pilot study with cognitively healthy OA in order to 1) investigate acceptability/usability and structure of a 12-session training protocol based on web B-Game, and 2) the effects of EmCh approach, in comparison to a control intervention. The K99 phase will provide pilot data regarding several intervention features and outcomes. These data will be critical to optimize the design for the randomized controlled clinical trial I plan to conduct in the R00 phase (Aim 3), in order to evaluate the efficacy of EmCh/B-Game in OA. The R00 trial will be enriched by Alzheimer`s disease blood-based biomarkers, which will improve diagnostic accuracy and allow me to explore intervention response as a function of pathology. To accomplish these aims, I will develop advanced skills to supplement my training in neuropsychology, as I will: 1) enrich my conceptual understanding of cognitive aging/EC; 2) broaden my knowledge of cognitive training, and its integration with technology and teleneuropsychology; 3) develop expertise in clinical trials; and 4) gain skills in statistical analysis. Additionally, I will 5) enrich my knowledge on Alzheimer`s pathology and 6) gain skills in designing fMRI experiments, relevant aims for my independence as a future trialist focused on aging. I have assembled an excellent and well-rounded mentorship team with expertise in cognitive aging, EC/EmCh, clinical trials, technological interventions, teleneuropsychology, and biomarkers. Overall, this K99/R00 proposal lays the foundation for an independent research career focused on technology-based intervention for OA, in order to reduce age-related cognitive decline and promote Alzheimer’s disease / dementia prevention.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Rapid escalations in substance use during adolescence confer risk for the development of substance and alcohol use disorders across the lifespan. Thus, identifying malleable risk factors and prevention targets that emerge before substance use onset is an important public health goal. Research demonstrates that delay discounting (DD), the tendency to perceive diminishing value in a reward as a function of the length of delay in its receipt, is an important and modifiable risk factor for substance misuse. However, the early developmental precursors of DD, including environmental adversity and genetic risk factors that are thought to be associated with increases in DD across adolescence are not well understood. The extant literature examining these vulnerabilities is limited by its reliance on poorly characterized markers of environmental adversity, lack of consideration of developmental effects, and the genetic contributions to the development of DD. The current application proposes to address these limitations utilizing data from the Adolescent Brian Cognitive Development (ABCD) Study, a national sample of youth (n = 11,875) with planned longitudinal assessments from age 9 to 17, spanning critical developmental periods for changes in DD and substance use. The proposed project will leverage rich and varied indicators of adverse childhood events and community environments (using the “Pair of ACEs” framework) and genetic data to create polygenic risk scores for DD (informed by findings from the largest available genome-wide association study of DD performed by project consultants). The project timeline will take advantage of both currently available and planned releases of data to test study aims and accelerate dissemination of findings from this landmark study. Specific aims of the study include: (Aim 1) to examine the structure of ACEs and its relation to early levels of DD; (Aim 2) to characterize the impact of ACEs on measured trajectories of DD over time; and (Aim 3) to investigate the joint associations between genetic and ACEs risk factors on changes in DD across adolescence. We also propose to explore whether developmental trajectories of DD mediate the relation between genetic and ACEs factors and subsequent changes in substance use frequency and severity. The research team brings together expertise in child development, delay discounting, genetics, and environmental indicators of risk, allowing for the most comprehensive study of the independent and joint contextual and genetic contributions to DD development. Findings from this project will provide mechanistic insights into the development of DD, in addition to specific and actionable advancements at the environmental level that may inform targeting of prevention approaches for reducing substance use among at-risk youth.

NIH Research Projects · FY 2025 · 2022-08

Project summary/Abstract SARS-CoV, MERS-CoV and SARS-CoV-2 belong to the genus Betacoronavirus and encode sets of specific accessory proteins. Accessory proteins encoded by coronaviruses are not essential for the viral life cycle but are important regulators that mediate immune evasion for optimal virus replication and propagation. One unique feature of Betacoronavirus that is not seen in other genera of the family Coronaviridae is the presence of a small accessory protein (I) encoded by the +1 open reading frame (ORF) relative to and within the ORF encoding the nucleocapsid (N) gene. The internal (I) proteins of SARS-CoV (ORF9b), MERS-CoV (ORF8b) and SARS-CoV- 2 (ORF9b) have not been extensively characterized. However, in vitro experiments suggest that the I proteins of these viruses have a role in suppressing IFN-I expression, which could potentially contribute to pathogenesis. In this application, we hypothesize that the I protein is a virulence factor with functions specific to each virus. The goal of this project is to study the roles of I proteins in pathogenesis and determine if I proteins possess functions specific to viruses within the genus Betacoronavirus. We generated mutant MERS-CoV and SARS-CoV-2 with deletions of I protein expression without altering the coding sequence of the N protein by reverse genetics and found that MERS-CoV lacking I protein expression showed increased virulence in mice, while the absence of the SARS-CoV-2 I protein resulted in attenuation. It is intriguing that the absence of the I protein resulted in such disparate changes in the virulence of the two related CoVs. In Aim 1, we will investigate the virus-specifc functions of the I proteins by inserting the I protein of MERS- and SARS-CoV-2 into mouse hepatitis virus (MHV), another betacoronavirus. In addition, we will interrogate the role of I proteins involved in regulating virus production by interfering with the virus/host machinery required for virus replication. This will be performed by analyzing the I protein interactome by mass spectrometry, detecting the presence of I protein in virions, which would be consistent with a role in virion assembly and release, and comparing the viral life cycle in mutant vs. parental virus-infected mice. In Aim 2, we will assess the role of the I protein in regulating immune responses by comparing mice infected with mutant viruses or parental viruses. We will interrogate the role of IFN-I signaling in the altered virulence of mutant viruses, as I proteins have been shown to suppress IFN-I induction in vitro. In addition, changes in immune responses will be investigated by measuring inflammatory cytokines in the blood, bronchoalveolar lavage (BAL) and the lungs of mice infected with mutant or parental viruses. Immune profiling of mice infected with mutant viruses will be performed and compared to mice infected with parental viruses by scRNA-seq. The training and experiments proposed in this career development award will not only offer invaluable opportunities for me to acquire new skills and techniques required for developing my own independent research in viral immunology and pathogenesis but also address important questions regarding how specific viral proteins act as virulence factors in coronavirus-infected cells.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Dementia with Lewy body (DLB) has been historically under-investigated relative to its prevalence as most studies of synucleinopathies focus on PD and PDD. Accumulating evidence indicates, however, DLB is a distinct age-associated neurodegenerative dementia. Like PD, various types of cells including neurons, microglia, astrocytes, oligodendrocytes, endothelial cells, and peripheral lymphocytes might contribute to DLB pathogenesis. To understand complexities of DLB pathogenesis, more comprehensive approaches to investigating different cell types and multiple brain regions over the course of disease progression are necessary. Our lab has put extensive effort into single-nuclei analysis of postmortem brain tissues using a recently released Chromium Single-Cell Multiome ATAC plus Gene Expression platform (10X Genomics) and successfully established all key techniques and a data analysis pipeline. To track the progression of disease using both RNA- and ATAC-seq data in the same cell, we have developed a novel strategy: “correlated pseudo-pathogenesis (cPP)” trajectory analysis. We also established “gene-peak” analysis allowing us to analyze relationships between gene expression and chromatin accessibility in a single cell. Our technical advancements and innovative data analysis skills will satisfy this FOA requesting “projects to identify cellular changes in ADRD post- mortem brain tissue across disease progression.” To achieve the goal set by the current FOA, we will pursue the following aims: Aim 1. Neuropathological staging of DLB and validation of control postmortem samples. Postmortem brain tissues from the HBTRC will be further validated for neuropathological staging by H&E and α- SYN immunohistochemical staining according to Unified Staging System for Lewy Body Disorders (USSLB). To eliminate incidental Lewy body disease (ILBD), control tissues will also be investigated for α-SYN pathology. Aim 2. Isolate nuclei from each sample and perform snRNA-seq and snATAC-seq analysis. 4 brain regions from three groups: 1) control; 2) stage II (limbic or brainstem predominant)/III (both limbic and brainstem); 3) and stage IV (neocortical), will be subject to single-nuclei multiomic analysis. Aim 3. Spatial transcriptomic analysis for topological domain mapping. Differential gene expression in the specific microdomains of each brain region and relationship to α-SYN pathology will be investigated using a spatially barcoded gene expression profiling array.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Human papillomavirus (HPV) causes over 34,800 cancers in the United States each year that disproportionately affect Black women. The HPV vaccine is highly effective, but less than half of adolescents aged 13-15 years have received recommended doses, with Blacks having the greatest disparity in completion. HPV vaccine hesitancy is associated with under-vaccination and refusal. An estimated 23% of parents with adolescents are hesitant about the HPV vaccine, but parents’ concerns vary due to a myriad of individual, interpersonal, and sociocultural factors. Effective strategies to overcome parents’ concerns are needed, especially among Black families, who have higher prevalence of general vaccine hesitancy and medical mistrust due to racial inequities. The purpose of this NCI Transition Career Development Award (K22) is to support me through specific training and research experiences as I become an independent investigator specializing in multilevel cancer prevention interventions to reduce cervical cancer disparities. Building on my strong health services research and behavioral science background in HPV screening, I will develop expertise in HPV vaccination communication, community- engaged intervention planning, and intervention trials. The objectives of this rigorous mixed methods proposal are to: (1) use a stakeholder-engaged approach to develop and refine an interactive, tailored text messaging intervention to address Black parents’ HPV vaccine hesitancy determinants and vaccination barriers; and (2) conduct a two-arm pilot RCT to determine feasibility, acceptability, appropriateness, and preliminary efficacy of the tailored messages compared to untailored messages. Following a provider’s initial recommendation, the intervention will allow parents of young adolescents to process HPV vaccination information on their preferred timeline, answer lingering questions and concerns, provide links to additional information from trusted sources, and support connections to local resources to help overcome barriers. By accomplishing these aims, I will address current gaps in strategies to increase vaccine confidence and motivation among high-risk Black families. Rutgers Cancer Institute with its well-funded behavioral research program, robust research infrastructure, and deep community connections is an exceptional environment to conduct high-impact, community-engaged cancer disparities research. As a logical next step in my career development, this K22 will give me the skills required to design, implement, and evaluate multilevel interventions to achieve HPV health equity. Findings and preliminary efficacy estimates will inform an R01 application of a multi-site trial to test a multiple component intervention addressing the complex, context-specific determinants of HPV vaccine hesitancy to motivate vaccination and change behaviors. Ultimately, this K22 proposal will facilitate my long-term goal to build an independent research program investigating community-driven solutions to reduce HPV disparities and advance the elimination of cervical cancer.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Despite dramatic improvements in Breast Cancer (BC) prognosis over the past two decades, major survival differences after diagnosis persist based on a number of clinical factors and tumor characteristics. However, even within similar molecular BC subtypes there are differences in survival, supporting that additional factors should be considered to better predict outcomes after diagnosis. Surprisingly, unlike risk models for first incident BC, current models for prediction of survival after a BC diagnosis and treatment do not incorporate host germline genetic variation. In addition, Black women experience a 40% higher mortality rate due to BC compared to their White counterparts. Further, they have been greatly underrepresented in genomic studies; so future clinical implementations of new precision medicine solutions based on germline genetic variation may exacerbate existing health disparities. This proposal aims to produce empirical evidence that will be an essential first step to improve BC prognosis based on appropriate clinical recommendations targeted to those with the highest risk of poor outcomes. In Aim 1a, I will investigate if a polygenic risk score (PRS) improves risk prediction of BC prognosis, over and beyond standard clinical markers and tumor characteristics in the Breast Cancer Family Registry (BCFR). Aim 1b will utilize the BCFR to examine the impact of adding a PRS to existing BC prognostic tools such as the Nottingham Prognostic Index (NPI), which incorporates information on tumor size, tumor grade, and lymph node involvement. In Aim 2a, I will examine if the PRS improves risk prediction for BC prognosis in the Women’s Circle of Health Follow-Up Study (WCHFS), a longitudinal study of Black BC survivors. Aim 2b will examine the impact of adding PRS to NPI using data from the WCHFS. My long-term goal is to translate epidemiologic findings into clinical care through more accurate risk assessment and risk-reducing strategies for outcomes after a cancer diagnosis. This K22 award will provide me with the necessary training and support to accomplish the following short-term goals: (1) obtain advanced skills in statistical genetics; (2) training in the translation of scientific research findings in the clinical context; and (3) professional development including learning the skills necessary to be a successful independent investigator. To achieve these goals, I have proposed a detailed career development plan, including taking short courses and workshops, attending national conferences, meetings with my advisory committee, and obtaining research experience by completing the proposed research aims. This K22 research will address critical knowledge and clinical translation gaps in identifying women who are the highest risk for poor BC prognosis. Given the increasing number of BC survivors and persisting BC survival differences for certain subgroups, this is a timely and important proposal.

NIH Research Projects · FY 2025 · 2022-08

Project Summary / Abstract Recent discoveries implicate specific genetic variants that confer extremely high risk for schizophrenia (SZ), a devastating psychiatric syndrome. Alongside these genetic discoveries there have been parallel advances in molecular neuroscience, including induced pluripotent stem (iPS) cell technology; high-throughput cellular technologies such as high content imaging and single cell genomics; and multiplex “cell village” approaches. These techniques allow for rigorous yet efficient interrogation of complex biological processes in previously inaccessible human neuronal cell types. The combination of genetic findings and technological advances are powerful tools for addressing what has become the “great white whale” of modern psychiatry: What is the underlying pathophysiology that gives rise to a SZ phenotype? We propose that high penetrance of rare SZ mutations derive from large effects at the molecular and cellular levels. We will identify downstream targets and pathways impacted by five rare SZ-associated variants with large effect sizes: deletions at chromosomal locations 2p16 (localized to the NRXN1 gene), 3q29, 15q13.3, 22q11.2, and duplication at 16p11. A key strength of this proposal is our access to the Genomic Psychiatry Cohort (GPC). Importantly, the GPC is a large cohort that is broadly representative of the US population. We will select for study previously-banked cryopreserved lymphocytes from individuals with SZ who carry one of these five defined variants (n=20 each genotype) to generate iPS cell lines. A clear advantage of the GPC is its large control sample, allowing us to select controls (n=40) that are matched by genomic background to the SZ cases, increasing the rigor of our study. A consistent but surprising observation about these SZ-associated rare variants is their similarity in both effect size and phenotypic characteristics, giving rise to the hypothesis that these variants converge on downstream molecular targets and/or cellular pathways. We will test the hypothesis that SZ-associated rare mutations cause molecular perturbations in neurons at the level of chromatin accessibility and gene expression and that genes or pathways impacted by two or more of these SZ-associated variants converge, with more overlap than expected by chance. Finally, we will validate molecular pathways using multimodal cellular phenotypic levels of analysis. Identifying the specific biological processes that are disrupted by SZ-associated loci will open a window into the complex molecular biology of this disorder. The substrate for our mechanistic studies will include subjects draw from the US population at large, ensuring that our results are generalizable to those suffering from adverse mental health outcomes. The tools and data generated herein will support the mental health field aligning with NIMH priorities, lead to transformative insights into the neurobiology of SZ, and uncover novel targets that may be a launch point for therapeutic discovery.

NIH Research Projects · FY 2026 · 2022-07

Project Summary/Abstract Central nervous system (CNS) control of metabolism plays a pivotal role in maintaining energy homeostasis. Glucagon-like peptide 1 (GLP-1, encoded by Gcg), secreted by a distinct population of neurons located within the Nucleus Tractus Solitarius, suppresses feeding. Central and peripheral GLP-1 work independently to suppress feeding . However, the cellular and circuit mechanisms mediating endogenous GLP-1 action in the CNS are still poorly understood. This is mainly due to the presence of diverse neuronal subtypes, complex central neuronal connectivity, and the lack of molecular tools that can directly detect GLP-1 release in the brain. Addressing the CNS mechanism of GLP-1 will help develop more tailored treatment for intervention of obesity. Our overarching goal is to gain a mechanistic understanding of endogenous GLP-1 release and its functions in the CNS in a cell type- and circuit-defined manner. In a previous study, we found that NTS GLP-1 projection to the paraventricular hypothalamic nucleus (PVN) enhances glutamatergic synaptic transmission, which is sufficient to suppress food intake, and ablation of PVN GLP-1R causes overeating and obesity. These results highlight the potential role of central GLP-1 in regulating energy homeostasis. However, GLP-1 signaling is complex due to the heterogeneity of PVN region GLP-1R neurons which form synapses with the dorsal motor nucleus of the vagus nerve (DMV) neurons and release glutamate, while also releasing g-aminobutyric-acid in the bed nucleus of stria terminalis (BNST). DMV and BNST may mediate food intake behavior differentially, i.e. homeostatic vs. hedonic feedings, but the roles that the PVN GLP-1R neurons-to-DMV and BNST projections play remains unexplored. Moreover, using our recently developed optical sensors for GLP-1, termed Reporter for Transmission mediated by G protein-coupled Receptor, we found the timing of GLP-1 release into the PVN is inversely related to eating bouts. We thus hypothesize that circuit and neuronal subtype-dependent endogenous GLP-1 signaling in the PVN regulates eating patterns (e.g. meal timing and sizes), energy expenditure, and food rewards. To test this hypothesis, we will determine the temporal dynamics of GLP-1 release and neuronal activity in the PVN during feeding episodes; and we will test the hypothesis that GLP-1 signaling regulates homeostatic and motivational feeding via different neuronal pathways. The results of this study will advance our conceptual understanding of the regulatory effects of endogenous GLP-1, facilitating the development of neuropeptide-targeting clinical interventions for eating disorders and obesity.

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Chronic infection by hepatitis B virus (HBV) is the leading cause of hepatocellular carcinoma (HCC). HBV alone is responsible for more than 3% of worldwide cancer deaths of all cancer sites combined. How HBV promotes carcinogenesis is not fully understood, leading to limited therapeutic options for advanced diseases. We previously revealed that 10% of HBV-associated HCC tumors carry clonal integrations of HBV in the KMT2B gene locus in cancer cell genomes. All these integrations are located between exons 3 and 6 of the KMT2B gene. KMT2B is the only close homolog of KMT2A (also known as MLL1). It has been well established that the chromosomal translocations affecting KMT2A are the primary drivers of infant mixed lineage leukemia. HBV integrations in KMT2B in HCC may mirror KMT2A rearrangements in leukemia, but its function has not been studied. We have identified patient-derived HCC cell lines carrying HBV integrations in KMT2B. Using these cell lines and RNA-seq data of HCC tumors with HBV integrations in KMT2B, we found that these integrations lead to expression of truncated KMT2B from the N-terminus to 697 to 906 amino acid residues. Knockout of the KMT2B truncation in an integration-carrying cell line suppressed cell proliferation in vitro. Overexpression of a KMT2B truncation transformed normal human hepatocytes in vitro and induced tumor growth in vivo in hydrodynamic injection-based mouse models. Mechanistically, we found the KMT2B truncation sequestered the tumor suppressor MENIN from the KMT2A/B histone methyltransferase complex and chromosome. A KMT2B truncation carrying a mutation that blocks MENIN binding failed to promote tumor formation in vivo. Based on these preliminary data, our central hypothesis is that HBV integrations in KMT2B produce truncations, which sequester MENIN from KMT2B complex and promote hepatocellular carcinoma. To test this hypothesis, we will determine the oncogenic function of KMT2B truncations, using various in vitro and in vivo models (Aim 1), and determine the mechanism of KMT2B truncation-triggered tumorigenesis (Aim2). Specifically, a new genetic engineered mouse model of HCC will be generated and a potential targeted agent for HCC with HBV integrations in KMT2B will be tested. The proposed studies aim to establish the oncogenic driving function and mechanism of HBV integrations in KMT2B in HCC, which cause 35,000 HCC cases every year. It is among the most common cancer-causing genetic alterations but it is significantly understudied. The completion of the proposed studies will discover a novel oncovirus-driving, epigenetic mechanism of HCC development and provide information for developing prevention approaches, early detection assays, and targeted therapeutics for affected individuals.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT The Food and Drug Administration (FDA) is considering a regulation that would reduce nicotine content in cigarettes to non-addictive levels. This could have profound implications for the health of the US population as cigarette smoking remains a leading cause of morbidity and mortality, and incurs nearly $170 billion in annual health care costs. An FDA analysis shows that a reduced nicotine cigarette policy could avert over 8 million premature deaths through 2100, but no study has evaluated its potential economic impact. It is also unknown how this policy could affect vulnerable populations, defined as groups at greater risk for poor health due to social factors or illness and disability. Clinical trials suggest that reduced nicotine cigarettes decrease smoking among vulnerable populations, but no study has evaluated their potential long-term implications. I plan to use simulation modeling methods to conduct health and economic impact analysis of a reduced nicotine cigarette policy for two vulnerable populations: 1) people with major depression, who continue to have higher smoking rates and less success with quitting compared to the general population; and 2) socioeconomically disadvantaged women of reproductive age, whose smoking increases risk for adverse maternal and infant health outcomes. I will extend a simulation model of smoking and depression that I previously developed to incorporate morbidity and economic outcomes, including direct medical costs and productivity losses. I will also develop a smoking model specific to women of reproductive age and level of educational attainment. This model will simulate smoking during pregnancy and its adverse effects on maternal and infant health, including premature delivery, low-birth-weight, small for gestational age, and sudden infant death syndrome. For both vulnerable populations, I will simulate the potential effects of e-cigarette use and an illicit market for normal nicotine cigarettes under a reduced nicotine cigarette policy. To be successful with this study and accurately assess the impact of the proposed regulation on costs to society, I need training in economic evaluation. I also need to deepen my understanding of the science of addiction and acquire background in reproductive health. To that end, I will enroll in formal coursework in cost- effectiveness analysis, addiction neurobiology, and reproductive health. I will also receive training from my team of mentors, who consist of NIH-funded investigators in tobacco regulatory science, health and economic modeling, behavioral pharmacology, and reproductive and mental health. At Yale, I will access a rich research environment through regular opportunities offered by the Tobacco Center of Regulatory Science, the Public Health Modeling Unit, and the Program in Addiction Medicine. This award will provide me with the skills and training I need to become a leading expert in health and economic modeling of addiction in vulnerable populations, and secure my path to career independence in tobacco regulatory science.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT There are significant knowledge gaps in understanding the mechanisms and biological predictors of low drug exposures and treatment failure in patients with tuberculosis (TB) and type 2 diabetes mellitus (DM). To address the impact of DM on poor TB outcomes, we propose an interdisciplinary mentored research and training plan to investigate gut microbiome-mediated variation of anti-TB drug pharmacokinetics (PK) in DM and non-DM TB patients in an ongoing prospective, observational PK trial investigating isoniazid, rifampin, and pyrazinamide in TB patients. Specific Aim 1 will quantify the effect of DM and gut microbiota alpha diversity on the bioavailability of oral anti-TB drugs in patients treated for active TB, by linking pharmacometric modeling with DM and measures of the gut dysbiosis in active TB patients with and without DM. Specific Aim 2 will characterize the relationship of gut microbiota alpha diversity and diabetes in patients with active TB, by conducting a comprehensive prospective analysis of the human gut microbiome from clinical stool specimens. Upon successful completion of the proposed K23 research, we expect our contribution to 1) establish the previously undescribed impact of the human gut microbiome as a significant covariate to explain the heterogeneity of drug PK in patients receiving active TB treatment and, 2) demonstrate the distinctive relationship between DM and gut microbiome diversity and composition among patients with TB. These contributions will be significant because they are expected to provide strong scientific justification for a novel mechanism for the previously unexplained variability of anti-TB drug PK and TB treatment response among patients with TB/DM. The proposed research is innovative because it aims to identify the gut microbiome as a novel mechanism underlying the heterogeneity of anti-TB drug PK. The overall goal of this K23 proposal is to train Navaneeth Narayanan, PharmD, MPH for a career as an independent investigator in pharmacomicrobiomics, the study of the effect of microbiome variation on therapeutic response by regulating drug PK and pharmacodynamics (PD), with a specific career emphasis on the treatment and outcomes of TB and other infectious diseases. The career development plan includes training in human microbiome research, under the mentorship of Dr. Martin Blaser, and pharmacometrics, an interdisciplinary science of quantitative clinical pharmacology and systems biology that involves expertise in mathematical modeling to characterize and predict drug PK and PD. Dr. Narayanan will also be mentored by Dr. Scott Heysell, an international expert in anti-TB pharmacology and TB clinical research. The proposed K23 project will provide an integrated plan of mentored patient-oriented research, career development activities, and formal training in microbiome research and pharmacometrics. Guided by expert mentors and collaborators, the research and training activities outlined in this application will enable Dr. Narayanan to mature into an independent clinical/translational researcher. These opportunities will equip this investigator with a larger set of skills to answer important and novel questions about global infectious diseases.

NIH Research Projects · FY 2025 · 2022-07

Abstract Cryptococcus neoformans and its sibling species C. gattii cause Cryptococcosis, a deadly fungal disease that accounts for over 15% of HIV/AIDS related deaths. Treatment options for cryptococcosis remain limited to two drug classes that are either highly toxic (polyenes) or exert a fungistatic effect (triazoles) that necessitate long treatment regimens and can induce drug resistance. The third antifungal drug class, echinocandins, shows low toxicity and is fungicidal against some prevalent fungal pathogens. However, Cryptococcus species are resistant to echinocandins through an unknown resistance mechanism. We found that loss of Cdc50, the regulatory subunit of lipid flippase, an enzyme that maintains asymmetry of the membrane lipid bilayers and regulates intracellular vesicle trafficking, sensitizes C. neoformans to the echinocandin drug caspofungin and several triazoles. We further showed that the cdc50∆ mutant abolishes lipid flippase activity. We also found that this Cdc50-mediated echinocandin resistance requires a mechanosensitive calcium channel protein, Crm1, which modulates intracellular calcium homeostasis. Strikingly, we discovered that lipid flippase function is essential for virulence in a murine model of cryptococcosis, suggesting that lipid flippase may be a novel antifungal drug target. In this project, our goals are to determine how lipid flippase mediates cryptococcal echinocandin resistance, and to conduct proof-of-principle studies of antibody-based inhibitors targeting flippase function as novel therapeutics for Cryptococcus infections. We hypothesize that C. neoformans has a unique plasma membrane structure and that loss of lipid flippase alters that structure to promote the interaction of caspofungin with its target and compromises fungal drug resistance mechanisms. We propose three Aims to test our hypothesis. In Aim 1, we will elucidate how loss of Cdc50 changes membrane structure to promote the interaction of caspofungin with its membrane target β-1,3-D-glucan synthase (Fks1). Aim 2 will identify the downstream drug resistance pathways that are compromised by the absence of Cdc50, which disrupts intracellular calcium homeostasis and promotes cell death. In Aim 3, we will develop an antibody Fab fragment and a stable peptide against the exoplasmic loop of Cdc50, which is essential for flippase function. We will validate how inhibitors sensitize C. neoformans to antifungal drugs and macrophage killing in vitro and in vivo in animal models. The region of Cdc50 targeted by this antibody-based approach has low sequence homology to its human counterpart, and our preliminary studies showed that an antibody raised against this region is fungal- specific, reducing the chance of off-target effects. The impact of this study to elucidate the mechanisms underlying lipid flippase mediated drug resistance in C. neoformans will be developing strategies for exploiting echinocandin drugs to effectively treat Cryptococci and other resistant fungal pathogens. Our successful development of antibody-based inhibitors will establish a new avenue of research and drug development against other membrane proteins in fungi and bacteria.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Preventing epilepsy and its progression (epileptogenesis) remains the ultimate goal for epilepsy research and therapy development. Although this has been identified as an urgent need by the NINDS Epilepsy Research Benchmarks, there is still no therapy available that interferes with the epileptogenic process. The development of a therapy to prevent epilepsy and its progression would be paradigm-shifting in the way epilepsy, one of the most frequent neurological conditions worldwide, would be treated. This application is designed to test new interventional therapies to prevent epilepsy in rodent models of acquired epilepsy utilizing existing FDA- approved drugs. The rationale for our approach is based on more than 25 years of research into maladaptive processes in adenosine metabolism, which drive, and contribute to, the epileptogenic process that turns a healthy brain into an epileptic brain. Specifically, an acute injury-associated adenosine surge in the brain drives glial activation and dysregulation of glutamate homeostasis through increased activation of adenosine A2A receptors, which couple to the astroglial glutamate transporter GLT-1. Hence, the use of the FDA approved A2A receptor blocker istradefylline, or the FDA approved GLT-1 activator ceftriaxone are rational therapeutic interventions to interfere with key mechanisms during the onset of the epileptogenic cascade. A delayed response of the epileptogenic cascade is pathological overexpression of the major adenosine metabolizing enzyme adenosine kinase (ADK) resulting in chronic adenosine deficiency in the epileptic brain. We have shown that overexpression of ADK, and specifically an isoform expressed in the cell nucleus (ADK-L), drives the epileptogenic process through an epigenetic mechanism (increased DNA methylation). We have shown that therapeutic adenosine augmentation effectively prevents epilepsy and its progression in 4 different rodent models of epileptogenesis. Hence ADK inhibitors and DNA methylation blockers (e.g. the FDA approved drug - 5-azacytidine) hold promise for the prevention of epilepsy and its progression. To this end we recently launched a drug discovery program, which yielded a candidate ADK-L inhibitor (MRS-4203) with antiepileptogenic activity. Collectively, our preliminary data provide a solid rationale that the adenosine system and its downstream mediators offer several possible antiepileptogenic therapeutic targets. The CENTRAL GOAL of this application is to identify and test therapeutic strategies for epilepsy prevention based on repurposing of FDA approved drugs and the use of novel small molecule compounds that target components of the adenosinergic system. Our therapeutic approaches will be tested and optimized in the mouse intrahippocampal kainic acid model of temporal lobe epilepsy and validated in a traumatic brain injury induced model of posttraumatic epilepsy. Our hypothesis will be addressed in three Specific Aims: (1) Targeting adenosine receptor dependent mechanisms for epilepsy prevention (2) Targeting adenosine receptor independent mechanisms for epilepsy prevention (3) Test therapeutic efficacy of antiepileptogenic combination therapies

NIH Research Projects · FY 2025 · 2022-06

Cancer cachexia (CC) is a systemic, metabolic wasting syndrome featuring body weight loss due to skeletal muscle and adipose tissue wasting. CC is suffered by ~80% of cancer patients that causes reduced performance status, intolerance to chemotherapy, and increased mortality. This debilitating condition is poorly understood and has no effective treatment. If CC therapy existed, it would improve treatment responses, increase quality of life, and prolong survival. With 50 years of study, the field has focused on defining pathways that promote atrophy in the end-organs most affected my cachexia. While this work has been fruitful, it has not led to identification of the upstream mediators of CC, nor has it generated effective therapies. There is an urgent need for high-quality discovery science and more detailed clinical phenotyping.We have created a virtual institute comprised of diverse, international, multidisciplinary scientists and clinicians with expertise in cancer, metabolism, neuroendocrine function, immunology, human metabolic diseases, preclinical models, and clinical phenotyping. We hypothesize that CC is driven by tumor-intrinsic factors that activate neurohormonal sickness pathways, which then induce anorexia, metabolic dysfunction, and tissue atrophy.

NIH Research Projects · FY 2026 · 2022-06

Project Summary The heart is a metabolically demanding organ, and derangements in metabolic processes lead to energetic deficits, generation of toxic metabolites, and redox imbalance, which drive pathogenesis to heart failure. However, current therapies for heart failure do not address this fundamental issue, and there remains an unmet clinical need for effective mechanism-based treatments. Altered transcriptional programs are thought to contribute significantly to impaired mitochondrial oxidative phosphorylation and insufficient energy production in heart failure. The estrogen-related receptors (ERRa, b, and g) are key regulators of mitochondrial respiration that are downregulated in the failing heart, yet the molecular mechanisms regulating their expression and activity remain largely unknown. In preliminary studies we have identified a novel molecular pathway that promotes the expression of ERRb, and ERRg in cardiomyocytes, and define a new heart failure pathway linking chronic stress to impairment of mitochondrial oxidative respiration. Our unexpected results demonstrate that the tumor suppressor protein Neurofibromin 2 (NF2) promotes proper metabolic function, and that cardiac deletion of NF2 predisposes the heart to pathological remodeling and failure in response to LV pressure overload stress. Transcriptome profiling of cardiac deficient NF2 cKO hearts indicated downregulation of metabolic pathways and decreased expression/activity of ERRb and ERRg. Using a proteomics-based approach, we identified the transcription factor Zscan21 as an interacting partner of NF2 and a novel positive regulator of metabolic gene expression and mitochondrial oxidative respiration in cardiomyocytes. Therefore, we hypothesize that endogenous NF2 engages the transcription factor Zscan21 to positively regulate expression of ERRb and ERRg and promote energy production during pressure overload stress in the heart. The objectives of the current application are to further define the clinical role of this pathway, and to elucidate the molecular mechanisms by which NF2 regulates expression of myocardial ERRb and ERRg and prevents energy deficit. These objectives will be accomplished in 3 aims. In Aim1, we will establish evidence of NF2 as an important and novel mediator of cardiac metabolic coupling and energy production during the initial and late phases of pressure overload stress. In Aim2, we will investigate in detail the molecular interaction between NF2 and Zscan21 and determine the ability of Zscan21 to regulate expression of ERRb and ERRg, mitochondrial oxidative respiration, and energy production in cardiomyocytes. In Aim3, we will determine the therapeutic potential of normalizing cardiac NF2 for treatment in the pressure overload model of HFrEF. The long-term objective of this project is to define mechanistic events that mediate mitochondrial metabolic dysfunction in heart failure and identify potential candidates for new therapeutic strategies targeting early stages of heart failure.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY Spiral ganglion neurons (SGNs), the primary afferent neurons in the cochlea, play vital functions in normal hearing by transmitting auditory information from the mechanosensory hair cells to the brain, and in restoration of hearing via cochlear implants in deaf individuals. However, exposure to traumatic and/or prolonged noise causes degeneration and subsequent loss of SGNs and their synaptic connections with hair cells in varied degrees, leading to degradation of auditory information, and impeding the performance of cochlear implants or future hair cell or synapse regeneration strategies. The reasons for such SGN degeneration remain unclear. To inform the development of novel therapies to preserve or regrow functional SGNs, it is critical to understand the biological mechanisms of SGN degeneration and survival in the injured cochlea. We have recently identified fractalkine signaling (CX3CL1-CX3CR1) between SGNs (which express chemokine CX3CL1 ligand) and innate- immune cells such as macrophages and monocytes (which express cognate CX3CR1 receptor) as a key neuroprotective signaling that promotes SGN survival and synapse repair in the injured cochlea. Here, we seek to examine the cellular and molecular mechanisms by which fractalkine signaling mediates neuroprotection in mouse cochleae following graded noise trauma. Specifically, Aim 1 will determine the precise roles of CX3CR1- expressing cochlear resident and blood-derived recruited macrophages in SGN survival or degeneration after noise trauma. Using fate mapping to distinguish and selectively deplete cochlear resident and recruited macrophages, we will test the hypothesis that CX3CR1-expressing recruited macrophages promote SGN survival after noise trauma. Aim 2 will determine whether CX3CR1 regulates macrophage responses after noise trauma such that absence of CX3CR1 results in an increased and sustained production of pro-inflammatory cytokines and reactive oxidative factors that is detrimental for SGN viability. Effector pro- and anti-inflammatory cytokines, and reactive oxygen and nitrogen species will be detected in both cochleae and macrophages with intact fractalkine signaling and those that lack CX3CR1 after noise trauma. Aim 3 will examine the relationship between human CX3CR1 polymorphisms and noise-induced hearing loss. Approximately 25-30% humans carry two single nucleotide polymorphisms (SNPs) in the CX3CR1 locus (hCX3CR1-I249/M280) that show defective binding to CX3CL1 ligand and loss of chemotactic function in macrophages. Using a novel humanized mouse model expressing the aforementioned human CX3CR1 SNPs, we will test the hypothesis that dysregulated macrophage responses due to impaired CX3CR1 signaling in these variants accelerates synapse and neuron loss and worsens hearing following noise trauma. Together, these studies will test fundamentally new hypotheses proposing specific elements of the innate immune system, macrophages and fractalkine signaling as critical targets for neuroprotective immunotherapies to promote synapse repair and SGN survival in an injured cochlea.

NIH Research Projects · FY 2026 · 2022-06

This is an application for an interdisciplinary project to develop a novel approach for personalized chemotherapy of gynecologic cancers. Estimates from the National Cancer Institute indicate that more than 116,000 women in the United States will be diagnosed with a gynecologic cancer and about 34,000 die from these types of cancer in 2021. Despite advances in surgical and radiation treatments, chemotherapy continues to be an important treatment option for gynecologic malignancies, especially for locally advanced and metastatic tumors. However, the efficacy of chemotherapy is substantially limited by the intrinsic and acquired resistance of cancer cells to cytotoxic drugs. We are proposing to develop and validate a nanotechnology-based approach of personalized treatment of ovarian carcinoma (most lethal type of gynecological cancers) constructed on the individual genetic profile of the patient’s tumor. Based on the results of the present translational research the following treatment protocol will be proposed for future clinical trials after the completion of the present project. Samples of a patient’s tumor and normal surrounding tissues will be obtained during the tumor debulking surgery and tumor profile data (the expression of predefined genes and proteins) will be obtained and analyzed. Based on this analysis, several molecular targets and the most effective anticancer drug(s) will be selected. Finally, a mixture of complex nanocarrier-based targeted delivery systems (TDS) containing drug(s)/siRNA(s)/targeted peptide will be selected from the pre-synthesized bank and the patient will be treated with the chosen cocktail of TDS designed specifically for their individual tumor. The selected systems will include the lipid-based carrier, the tumor targeting moiety, the most effective drug(s) and siRNA(s) selected for each individual patient based on a genetic profile of the patient’s tumor. It is expected that such personalized therapy will effectively suppress drug resistance and tumor growth, inhibit the development of metastases and limit adverse side effects of therapy in the particular patient. The main goals of the proposed research are to identify profiles of gene/protein expression in tissue samples isolated from patients with ovarian cancer that predict tumor response and resistance to anticancer drugs with different mechanisms of action. We also will develop a set of TDSs containing anticancer drug(s) or siRNA(s) targeted to different mRNAs overexpressed in the tumor of the patient. Finally, a genetic profile and protein expression phenotype will be performed on samples of tumor tissues and malignant ascites from patients with ovarian carcinoma. Cancer cells will be isolated from fresh samples obtained during surgery. Based on the results of the genetic profiling, a mixture of TDS will be created and tested in vitro (on cell culture model) and in vivo (on subcutaneous murine cancer model) using cancer cells isolated from each individual patient and recommendations for the personalized treatment of ovarian cancer will be developed.

NIH Research Projects · FY 2024 · 2022-06

Project Summary Perceiving multisensory information and responding with appropriate, real-time behaviors is critical for normal communication and interaction with the environment. Past studies have investigated general brain regions as well as specific cells that fire in response to more than one type of sensory cue, yet have not pinpointed their presynaptic unimodal partners (inputs), meaning that the studied cells may have not been the direct points of multisensory convergence. Additionally, these neurons themselves did not then drive responsive and innate motor behavior. This has left gaps in understanding (1) how multisensory neurons acquire their multi-modal feature detection properties directly from unimodal inputs at the cellular and circuit levels, (2) how they integrate multimodal signals over time, and (3) how they then transform those signals into dynamic motor responses, all during ethologically relevant and innate interactions. Neural circuits that govern Drosophila melanogaster courtship serve as a well-developed model for sensory processing and real-time behavioral responses: during fly courtship, males sing to females, and females perceive and respond to song, which in turn alters males’ own courting behavior. This forms a complex “conversation” that emulates properties of many animals’ social interactions. Within the courtship circuit, I have discovered two direct unisensory convergence points onto multicellular cells and circuits, which have themselves been shown to be necessary and sufficient to drive and modulate robust, measurable, and innate behavior in Drosophila females, providing an unprecedented opportunity to address the gaps described above. The convergence points were identified via analysis of novel whole-brain and half-brain electron microscopy datasets at synaptic resolution. This proposal will directly elucidate principles of multisensory integration at the cellular, circuit, and behavioral levels. Through high resolution behavioral tracking assays, Aim 1 will determine how, by integrating natural combinations of audiovisual information, two multisensory neurons drive an ethologically relevant behavior in Drosophila females. Aim 2 will determine, using calcium imaging in behaving flies in a courtship virtual reality, how those neurons integrate auditory and visual signals from three of their unisensory inputs. Taken together, this study will significantly expand understanding of how brain cells and circuits process multisensory signals and transform them into dynamic motor responses, contributing to a foundation for long-term understanding of normal and disordered sensorimotor function. Additionally, this scientific proposal, along with the outstanding training environment in the Murthy lab and at the joint MD/PhD program at Princeton and Rutgers, will provide exceptional foundational training in preparation for my career as an independent physician scientist with my own laboratory.