Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT: In the last 50 years, over 150 types of congenital disorders of glycosylation (CDG) have been described, while available treatment options have severely lagged. CDG are early onset childhood diseases that often present with severe symptoms that span most organ systems. Collectively, heterogeneity in the genetic causes and clinical presentations of CDG have made treatment challenging, and thus, there is a significant need to elucidate the mechanisms of individual CDG to provide available treatment options for patients. One CDG with no available treatment and often severe symptoms is RFT1-CDG, caused by mutations in the transmembrane protein RFT1. RFT1 is proposed to function in N-glycosylation and/or GPI-anchoring, however, its role in glycosylation remains undefined. Many RFT1-CDG symptoms are related to defects in growth and development. My preliminary work in C. elegans suggests that RFTH-1 (RFT1 homolog) may activate Hedgehog signaling, a key regulator of development and growth, and that downstream signaling occurs through the pro-growth mTOR Complex 2 (mTORC2) pathway. Glycosylation can dramatically alter protein function or activity, and interestingly, multiple Hedgehog signaling components are glycosylated. Thus, impaired developmental and pro-growth signaling may underlie RFT1-CDG pathophysiology. This proposal seeks to investigate how RFT1 governs development and growth via regulation of Hedgehog and downstream homeostatic signaling, as well as to deconvolute the role of RFT1 in the glycosylation of Hedgehog signaling factors and establish how these events alter organismal physiology. This work will be conducted using genetic, cell biological, and biochemical approaches in two complementary model systems: C. elegans, to gain a whole organism perspective, and mammalian cell culture, with the goal of translating our C. elegans work to understand the disease pathophysiology in humans. In doing so, I aim to elucidate the mechanisms by which RFT1 mutations cause CDG-associated symptoms, while establishing a framework to study other CDG types to inform the development of effective treatment options. This research will take place at the University of North Carolina at Chapel Hill (UNC-CH) in the Rob Dowen Lab as part of the Genetics and Molecular Biology (GMB) Curriculum. UNC-GMB upholds a collaborative, innovative, and rigorous research program with excellent resources, outstanding facilities, and a substantiated commitment to trainee success. In addition to building my scientific skillset through this proposed work, my training will include strengthening my scientific communication and leadership skills to prepare for a career as a principal investigator. Through GMB and its affiliated career development office, TIBBS, I will have abundant resources to hone these skills. Additionally, my dedicated, interdisciplinary team of mentors, including my advisor and thesis committee, will support both my scientific and career development training endeavors. Altogether, my scientific environment and training plan will equip me to execute this proposal and become a well-rounded, independent scientist.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Since its inception in 2012, the UJMT Fogarty Global Health Fellows Program—led by the University of North Carolina at Chapel Hill, Johns Hopkins University, Morehouse School of Medicine, and Tulane University—has provided a critical pathway to global health research careers. Our 252 trainees have collectively secured $88.5 million in grant funding, a nearly 600% return on the NIH’s investment. During the current grant cycle (2017- present), our 110 doctoral scholars and postdoctoral fellows have published a collective 287 manuscripts, including 109 as primary author. The vast majority remain involved in global health research, and many have already obtained independent grant funding. In the coming grant cycle (2022-2027), under the new LAUNCH initiative, we build upon these strengths to further improve and expand our program. First, we will provide enriching 12-month mentored training opportunities at affiliated research sites in 16 countries. We draw upon a highly qualified group of US and international researchers jointly recruited from 12 domestic and 17 international training programs funded by the NIH. Our expert mentors support experiential learning on site, while the program provides a central training framework that monitors progress through individual development plans, core skills checklists, and regular group check-ins. Over the course of the five-year award, we expect to support at least 60 positions via base funding; in line with the RFA, 60% will be US postdoctoral fellows. Second, we will continue to foster research in scientific areas identified by in-country stakeholders. This includes topics described in the RFA, as well as evolving federal and NIH priorities. Third, we will enhance our training curriculum. We have developed a new Research Leadership Academy for Global Health, building upon our team’s experience in global health and leadership, institutional resources, and collaborations with other US government programs. We also provide individually guided research methodology training and scientific writing support, using proven approaches. Fourth, we will monitor, evaluate, and learn from our activities to iteratively improve the quality of our trainee experience. We utilize a conceptual framework that considers both process and performance to ensure that the specific aims of the program are met. Finally, the UJMT LAUNCH initiative will be led by a team of committed global health researchers—and a broad network of engaged collaborators, both in the US and abroad—with the expertise and experience to carry out its ambitious aims. Through this program, we will train a group of productive, independently funded, research leaders who are prepared to tackle the most pressing global health issues in the years to come.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Cardiovascular disease (CVD) is one of the leading causes of death in the US that disproportionally affects minorities and low-income individuals. The risk for and course of CVD is affected by social factors such as daily stressors (i.e., interpersonal conflicts and demands). Daily stressors and strong reactivity to stressors (stronger affective responses to daily stress) are strong predictors of CVD and two risk factors for CVD: higher cardiometabolic risk and inflammation. One central source of daily stress during midlife is parenting, and many parents and their children report increased stress during adolescence. The daily stressors of adolescents can crossover to affect the health of their parents. Our preliminary studies have found that on days when adolescents experience stressors, their parents are more likely to experience negative affect, physical symptoms (i.e., headaches, flu-like symptoms), and unhealthy physiological responses to stress (i.e., high cortisol) that same day. However, little is known about how adolescent stress can crossover to affect their parents’ long-term CVD risk. Such knowledge is critical given that adolescent stress may be a driver of poor parent health and developing interventions to interrupt crossover processes may be an important avenue to reduce parents’ risk for CVD. In this study, we test the associations between adolescent daily stressors and two risk factors for their parents’ CVD: parent cardiometabolic risk and inflammation. We use three unique samples of parent-child dyads (n=718) with different demographic profiles to (a) test if our findings are replicable on different populations and (b) identify important socioeconomic (SES), racial, ethnic, and sex disparities in the crossover effects of adolescent stress on parents’ CVD risk. Using both secondary data and new primary data collection, in Aim 1 we identify the crossover effects of adolescent stress on parent health by testing whether (a) adolescent daily stressors (stressor exposure, severity, emotional responses) and (b) parent reactivity to adolescent stressors predict parent cardiometabolic risk and inflammation concurrently and longitudinally. In Aim 2 we identify the psycho-cognitive (psychological distress, worry, perseverative thinking), social (parent-child warmth and conflict, parent loneliness), and behavioral factors (sleep) that mediate the crossover effects of adolescent daily stressors on parent cardiometabolic risk and inflammation. In Aim 3 we identify health disparities by testing whether the associations tested in Aims 1 and 2 are moderated by parent socioeconomic status, race, ethnicity, and adolescent biological sex. Public Health Impact: This project will identify (a) how adolescent stress impacts parent cardiometabolic risk and inflammation, (b) the modifiable psycho-cognitive, social, and behavioral mechanisms underlying these associations, and (c) socioeconomic, racial, ethnic, and sex disparities in these processes. Findings will lead to the design of a future parenting intervention to help parents respond to adolescent stress in ways that promote parent health.

NIH Research Projects · FY 2025 · 2025-09

Abstract Alzheimer’s disease (AD) is characterized by the propagation of tau aggregates throughout the brain. Tremendous efforts have been made to the development of tau biomarker, aimed at advancing our current understanding of AD etiology and facilitating early diagnosis in routine clinical practice. Since current pathology imaging technologies only provide a spatial mapping of tau pathology, many biomarker studies have to link stereotypical patterns of tau accumulation to the complex pathophysiological mechanism of disease progression. Recently, we have made significant progress to characterize dynamic process of tau spreading in a principled potential energy transport model that allows us to uncover the region-to-region spreading pathways of tau aggregates from the longitudinal tau-PET scans. However, our current method is limited to a single-layer scenario to the extent that the spreading pathways follow either the wiring topology of white matter fiber bundles (referred to as "air-based") or the folding patterns on the brain cortex (referred to as "road-based"), despite the fact that air-based and road-based transport might collaboratively contribute to the whole-brain tau propagation. To address this limitation, we will develop a multi-layer neural transport equation to jointly model the spreading of tau aggregates throughout the brain network and along the brain cortex in Aim 1. Furthermore, we introduce the notion of optimal control to characterize the interaction between air-based and road-based tau propagation, which allows us to uncover a system-level understanding of the selective mechanism in multi-layer tau propagation in Aim 2. By capitalizing on large scale of existing neuroimaging data, we will explore a set of open neuroscience questions regarding tau propagation in Aim 2, including dynamic coupling patterns, region vulnerability, and system criticality. Meanwhile, we will evaluate the clinic value of our multi-layer model in predicting the future tau accumulation and the risk of developing AD. The success of this project has the potential to offer valuable insights into AD etiology, thereby enhancing our understanding of AD progression and refining treatment strategies. Moreover, the computational tools can be readily applied to other biomarker studies in neurodegenerative diseases. This initiative lays the groundwork for future research into advanced diagnosis and tailored personalized treatment for AD.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Our research develops new microscopes that transform our ability to observe biological specimens and applies these microscopes to solve fundamental problems in cell biology. Light microscopes are a cornerstone of biological research. Recent optical, technological, and computational advances have greatly increased the resolution and speed at which we can observe living samples. However, in many cases, the ability of instruments to generate data has greatly outpaced our ability to analyze, interpret, and act upon this information. This is especially true for microscopes that can image across spatiotemporal scales, observing single molecules to whole organisms. Over the next five years, we will combine new optical principles together with computer vision to build autonomous microscopes that can not only capture data more rapidly, but also more efficiently. These instruments will be able to process data in real time and use this information to navigate through and interact with biological specimens based on experimental goals. In parallel projects, we will leverage recent advances in light sheet and single molecule imaging to study a key biological question, namely, how thousands of unique proteins navigate within and accomplish distinct biological functions in the cell nucleus. The overall vision for this research is to develop new microscope capabilities that enable new studies of biological processes and to test key hypotheses about how functional environments are organized in the nucleus of individual cells.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Sickle cell disease (SCD) is a complex and highly morbid inherited blood disorder that affects at least 100,000 people in the US. It is associated with a 20-year reduction in life expectancy and substantial economic costs to the health care system. Research and clinical care for SCD has faced challenges from a lack of population- level registry data and a historic paucity of specialist care relative to comparator inherited diseases like hemophilia or cystic fibrosis. Recent years have seen an increased focus on the development of comprehensive sickle cell centers (CSCCs); however, we do not know how many people with SCD have access to CSCCs at a population level, nor do we know the impact of receiving care at these centers on health-related outcomes. To address these gaps in knowledge, we propose two analytic aims using national insurance claims data from Medicaid, which insures the majority of people with SCD in the US. In the first aim, we will identify three groups of people with SCD: those who A) do not receive care from a hematology provider, B) receive care from a hematology provider but not at a CSCC, and C) receive health care at a CSCC, and we will identify sociodemographic characteristics associated with not receiving care at a CSCC. The second aim will use an econometric approach called instrumental variable (IV) analysis, which is a quasi-experimental design that accounts for bias in observational data to produce estimates of causal effect. We will use IV analysis to estimate the causal effect of receiving care at a CSCC on mortality and health care utilization, using differential distance to a CSCC as an instrument. The proposed research will provide the first national description of access to comprehensive care for people with SCD and the most robust estimate of the impact of receiving comprehensive care to date. These results will help health care providers consider the role of CSCCs for people with SCD, and they will inform policymakers’ discussions about allocation of resources toward comprehensive care in SCD. This work will also provide critical training to the applicant in the form of mentorship in SCD and advanced research methodology as he pursues his goal of becoming an independent health services researcher in SCD.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Metabolic programming is a critical regulator of inflammatory responses. Prior work has shown that changes in metabolism drive changes in macrophage activity. These metabolic changes are not only a consequence of polarization but also drive macrophage function. Downregulation of fatty acid oxidation within pro-inflammatory macrophages leads to enhanced lipid synthesis, including production of pro-inflammatory lipid mediators. The majority of these studies have been performed in bone marrow-derived macrophages (BMDM), and questions remain as to how metabolism regulates the activity of lung macrophages during influenza A virus (IAV) infection. Within the lungs, macrophages play a critical role in host defense and mediate lung inflammation and resolution. Lung macrophages can be classified as tissue resident alveolar macrophages (AM), interstitial macrophages, and monocyte-derived macrophages. Recent work has demonstrated significant differences in the metabolism of AM and BMDM. Unlike BMDM, pro-inflammatory AM do not upregulate glycolysis, and in fact upregulation of glycolysis is associated with a pro-repair or pro-fibrotic phenotype. Our preliminary data indicate that there are significant differences between AM and BMDM not only in glycolysis but also in lipid metabolism. AM have a higher concentration of metabolites involved in fatty acid oxidation and have higher mRNA expression of the enzymes Cpt1a and Cpt2, which are rate limiting enzymes for fatty acid oxidation that act sequentially to transport fatty acids from the cytosol into the mitochondrial matrix where fatty acid oxidation occurs. In addition to serving as a source for ATP, lipid metabolism also generates lipid mediators that have important roles in cell signaling, cytokine production, and apoptosis. Thus, our proposal focuses on understanding how changes in lipid metabolism alter the activity and function of lung macrophages during homeostasis and injury. Our hypothesis is that fatty acid oxidation is critical for resolution of lung injury by modulating the activity of lung macrophages and reducing production of pro-inflammatory lipid mediators. In Aim 1, we will determine the role of fatty acid oxidation on lung macrophage activity during IAV infection. We will utilize LysM-Cre Cpt1afl/fl mice, which have a macrophage-specific deficiency in fatty acid oxidation. After inoculation with IAV, the clinical and inflammatory responses and severity of lung injury will be assessed at multiple time points. Lung macrophage subsets will be isolated and their transcriptomes profiled by RNAseq. In Aim 2, we will determine how fatty acid oxidation regulates the metabolic activity of lung macrophages by performing unbiased metabolomics on isolated lung macrophage subsets during IAV infection. The extracellular microenvironment will be assessed by determining the metabolome of the bronchoalveolar lavage fluid. These studies build upon work done in my K08 award and expand our focus to fatty acid oxidation and lipid metabolism. These data will serve as a foundation for an R01 proposal aimed at understanding how changes in metabolism regulate lung macrophage activity during injury.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Four rare bronchiectatic diseases (RBDs), including primary ciliary dyskinesia, alpha-1 antitrypsin, humoral immune deficiency, and STAT3-HIES, manifest bronchiectasis (BE) as a severe pulmonary complication. BE, defined by airways dilation/ectasia on chest CT imaging, is associated with cough, sputum production, acute exacerbations (AE), loss of lung function, and increased mortality. Progress in therapeutics for the most prevalent form of BE (non-cystic fibrosis bronchiectasis, NCFB), and by implication, the 4 RBDs selected for study, has been slow. Part of this “tardiness” in therapeutics development has been the incomplete nature of the “Vicious Vortex” hypothesis, a paradigm for investigating BE pathogenesis which we posit is missing key elements that initiate and perpetuate the “Vortex”. Presently, there is an urgent need to accelerate BE therapeutics development. Key missing elements include data describing the clinical trajectory of each RBD and disease- informative BE biomarkers. Using new insights and biomarkers derived from a novel “Vicious Vortex 2.0” hypothesis, combined with new observational studies, this U54 proposes to fill in the gaps to accelerate BE drug discovery, build a pipeline of rare disease investigators, and conduct ancillary studies, some in partnership with industry. This work will be conducted in partnership with the Patient Advocacy Groups (PAGs) associated with the 4 RBDs. To accomplish these goals, we propose 2 projects: Project 1, “Longitudinal Study of Clinical Outcome Measures and Biomarkers in Rare Bronchiectatic Diseases” (K. Olivier, MD, MPH, PI), will define clinical trajectories of the 4 RBDs over a 3-year longitudinal observational period as quantitated by pulmonary function tests, patient-reported outcomes, AEs, and novel CT analyses. Ancillary goals are to collect sputum, BAL, oral, and blood samples for biomarker testing. Emphasis for biomarker development is focused on novel mucus parameters, anaerobic microbiomes, and metabolomics. Airway epithelial and microbiome samples will be collected for collaborative studies with Project 2. Project 2, “Rare Bronchiectatic Disease Biomarkers and Novel Treatable Traits” (R. Boucher, MD, PI), provides complementary laboratory-based studies of patient- derived materials obtained from Project 1, supplemented by studies of excised lung specimens from the 4 RBDs. Goals are to utilize high-content RNA/protein technologies, coupled with studies of cultured airway epithelia from RBD patients designed to measure responses to BE provocants (aspiration, viral infections, and oral microbiome challenge) to identify/validate novel biomarkers and treatable traits. Cores focused on Career Enhancement, Pilot and Feasibility Study Development, and Administration have been assembled to achieve our goals. Deliverables will be: 1) high-quality/high-content clinical data describing the 3-year clinical trajectory of the 4 RBDs, coupled to identification/validation of a suite of novel biomarkers vetted by in vitro and in vivo studies; 2) identification of novel RBD common and specific “treatable traits”; 3) training the next generation of rare disease investigators; and 4) conducting ancillary studies that will lead to clinical trials.

NIH Research Projects · FY 2025 · 2025-09

Cardiovascular disease is not equally distributed across ethnic and racial groups. Studies have shown that between and within-group disparities exist due to differences in social and economic positioning and related experiences. However, one underexplored area in social positioning is examining disparities based on demographic characteristics. For example, research across the Americas publish that different social categories result in a varied cardiovascular outcomes and wellbeing than others. Although all individuals embody both ethnicity and a race, for instance, research often flattens these varied experiences in favor of a single self-identification. How individuals self-identify is neither a linear process nor just influenced by individual-level factors. Different socialization experiences can produce differential exposures to risk thus producing distinct cardiovascular outcomes. For different communities, having robust measures of demographic variables remains as a gap. For instance, different communities such as Latinos, have high levels of missingness when using standard self-reported demographic questions, thus hindering our ability to investigate health disparities. This R03 study is informed by socialization and self-identity theories and uses qualitative and quantitative methods to begin addressing this gap in the literature and relatedly, our processes in data collection. This R03 proposal leverages results from an existing K01 study exploring self-identity among multiracial individuals and gaps on existing measurement of demographic variables to: (1) develop a multi-item demographic variable survey module, (2) evaluate the association between each demographic variable and cardiovascular risk factors, and (3) establish guidance on measuring demographics for different subgroups participating in research. For Aim 1, the study team will develop a demographic variable survey module comprised of existing items (i.e., questions) and novel items informed by the K01 study. The team will use cognitive interviews to assess the content validity of demographic variable survey module. For Aim 2, the study team will integrate the demographic variable survey module into a cross-sectional survey of cardiovascular risk factors for distribution to adults in the United States to assesses differences in cardiovascular risk across subgroups. For Aim 3, the study team will convene an advisory board to utilize R03 study results and existent literature to formulate guidance on best practices for using demographic questions in public health and medical research. The goals of this R03 study are consistent with the NHLBI’s strategic goals: (1) to investigate factors that account for differences in health among populations and (2) optimize clinical and implementation research to improve health and reduce disease. This proof-of-principle study will produce preliminary data on CVD disparities affecting US adults through the testing of a novel demographic variable survey module, which will inform tailored research studies to reduce health disparities between and within subgroups.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Significance: Access to insulin, a biologic drug, has become a burden for many patients due to high prices. Biosimilars offer a solution to overcome this burden. They are biologics that are highly similar to the originator- biologic but not an exact copy. They are designed to match the structure and clinical effects of the originator- biologic. In July 2021, insulin glargine-yfgn (Semglee) became the first interchangeable biosimilar insulin approved. However, the randomized trials used to support the approval of Semglee were small in sample size, short in duration, and lacked diversity. Thus, there is an urgent need for real-world evidence on the use and safety of Semglee compare the originator-biologic insulin glargine (Lantus). Specific Aims: The proposed project will provide valuable information to patients and stakeholders on the use and safety of Semglee in a real-world population and demonstrate novel pharmacoepidemiology methods to evaluate the safety and interchangeability of biosimilar insulins post-market approval. In Aim 1, we will quantify and characterize the switching patterns (e.g., Lantus-Semglee-Lantus) of patients who received a prescription for Semglee or Lantus and assess their characteristics to understand how patient characteristics influence biologic and biosimilar insulin use. In Aims 2 and 3, we will use novel pharmacoepidemiology methods to replicate and emulate the randomized trials used to support the approval of Semglee with real-world data and compare the rate of serious hypoglycemia between Semglee and Lantus. Approach: This study will compare the use and safety of Semglee to Lantus with real-world data from the Merative MarketScan database, a longitudinal administrative claims database of more than 250 million private insurance beneficiaries. In Aim 1, we will identify switching patterns (e.g., Lantus-Semglee-Lantus) among patients who received a prescription for Semglee or Lantus and assess their demographics and clinical characteristics at baseline, `time of first switch', and `time of first switchback'. In Aim 2, we will compare the rate of serious hypoglycemia between Semglee and Lantus over a 1-year follow-up period with the prevalent new- user study design. In Aim 3, we will emulate a randomized switching trial to compare the interchangeability (effect of switching) between Semglee and Lantus to continuous (non-switching) Lantus on the rate of serious hypoglycemia over a 9-month follow-up period with the clone-censor-weight approach. Fellowship Information: The principal investigator is a PhD student in Epidemiology at UNC. Dr. Her proposes a training plan that will equip him with the knowledge and skills to launch a career as an independently funded principal investigator in diabetes. His training plan consists of mentor research, coursework and seminar, and professional development that will take place in a thriving collaborative research environment, guided by an internationally recognized team of mentors.

NIH Research Projects · FY 2025 · 2025-09

Injury-related fear after anterior cruciate ligament (ACL) injury significantly contributes to decreased return to sport, decreased physical activity engagement, and increased secondary ACL injury in previously high functioning, physically active individuals. Injury-related fear is also associated with poor jump-landing movement patterns and worse self-reported knee function in patients after ACL reconstruction (ACLR). While injury-related fear can be modified using psychological interventions, we have not comprehensively characterized how, when, and what types of injury-related fears (e.g., fear of movement, reinjury, avoidance) impede recovery. Furthermore, there is limited evidence regarding patient perceptions of injury-related fear and their perceived consequences on recovery outcomes. Failure to identify what types and when these fears are present throughout ACLR rehabilitation, and how patients perceive these fears to impact their recovery, will significantly inhibit our ability to effectively intervene with robust and effective psychological interventions aimed to mitigate secondary ACL injury risk. We hypothesize that patients follow three separate trajectories, including favorable (i.e., decreases in fear that are maintained), unfavorable (i.e., unresolved fear at clearance for return for activity), and variable trajectories (i.e., decreases in fear likely due to direct intervention prior to return to activity). Identifying these trajectories will enhance intervention effectiveness for future clinical trials using psychological interventions to mitigate secondary ACL injury risk and improve long-term knee joint health. Consequently, there is a critical need for high-quality longitudinal studies to characterize injury-related fear, both quantitatively and qualitatively, in patients after ACLR. Therefore, the objective of this study is to characterize injury-related fear in patients post-ACLR. A total of 50 patients will complete surveys to assess injury-related fear at important clinical timepoints, including pre-operatively, and at 2-, 4-, 6-, and 9-months post-ACLR. A subset of 15 patients at 9- months post-ACLR will complete semi-structured interviews to identify patient perceived impacts of injury-related fear on recovery outcomes and discuss interventions that could be used to reduce injury-related fear after ACLR. The central hypothesis is that multiple trajectories will exist for injury-related fears (i.e., fear of movement, fear of reinjury, and fear-avoidance beliefs) which will be supported qualitatively through patient interviews. The findings from this proposal will address a gap in knowledge by providing a clear understanding of how injury- related fear changes over time in patients after ACLR. This proposal will also position an aspiring independent clinician scientist with pertinent data that is needed to support future NIH grant applications and the trajectory of her independent research career.

NIH Research Projects · FY 2025 · 2025-09

Abstract Alzheimer’s Disease (AD) has been identified as one of the highest priority neurobiological diseases in need of research advancement, due to the progressive memory and emotional impairments inflicted on patients, and the excessive burden for caregivers and families. Due to the continued lack of effective preventative treatments for AD, it is imperative to identify ways to remediate the cognitive and affective impairments associated with the disease, and it appears that a more synaptic and circuity focused approach may be necessary. The hippocampus is one of the earliest regions in human and animal models to present with AD pathology and synaptic disfunction, and it has established roles in both memory and emotional regulation. The dentate gyrus (DG), in particular, undergoes many important circuitry and excitability changes in human cases of AD. As the first stop in the tri- synaptic loop, DG has significant control over downstream activation of the CA3 and CA1 regions of the hippocampus. Furthermore, since CA1 and subiculum project to many other structures throughout the brain, important for memory consolidation and animal behavior, activity balance in the DG may impart brain-wide functional connectivity changes under pathological conditions. Our previous research found that glutamatergic Mossy Cells (MCs) in the DG have the unique capability to recruit either excitatory or inhibitory neurons depending on the extent of MC activation. More recently, it has been discovered that MCs play a much more significant role in hippocampal circuitry than previously thought, as these cells have unique anatomical projection patterns that not only cross to the contralateral hemisphere, but also project longitudinally, along the entire dorsal-ventral axis of the hippocampus. Our preliminary data acquired from the 5xFAD rodent model of AD has identified a specific decrease in activity states of the dorsal population of MCs by 4.5 months of age, and a loss in overall DG granule cell activity. Because of the unique anatomical characteristics of hippocampal MCs, and due to the potential of dorsal MCs to regulate activity of DG granule cells throughout the hippocampus, we postulate that loss of dorsal MC signaling is one of the key circuit deficiencies contributing to cognitive and affective deficiencies in AD. By utilizing activity-dependent stimulation of dorsally targeted populations of MCs, we aim to characterize the breakdown of excitatory and inhibitory control throughout the different regions of the diseased hippocampus. We will use a combination of in vivo calcium imaging, chemogenetic stimulation of dorsal MCs, small animal behavioral analysis, and awake small-animal fMRI scanning in mouse models of AD to examine the effect MCs have on hippocampal activity and functional connectivity throughout the brain. Identification of discrete network components with the potential to compensate for improper excitatory/inhibitory balance and connectivity will greatly inform on the potential for circuity-based interventions to address the cognitive and affective impairments of AD.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Cardiovascular disease (CVD) is the leading cause of death in the United States, yet drivers of disease and risk markers remain poorly characterized. Although CVD is more prevalent among non-Hispanic Blacks and Hispanics compared to non-Hispanic Whites, these populations are underrepresented in existing genetic and epidemiological studies of disease. New affinity-based high-throughput proteomics platforms offer unprecedented opportunities to study disease mechanisms across diverse populations, and to search for new clinical biomarkers of disease. However, initial comparisons of the leading proteomics platforms suggest that at least one-third of protein measures correlate poorly between platforms, resulting in platform-specific or platform-discordant genetic and epidemiological associations, which may confound biological interpretation of results across studies. Genetic variants associated with circulating protein abundance measures may highlight assays with on-target affinity-probe binding, as well as assays which may be non-specific or prone to interference from protein-altering variants, which may affect affinity-probe binding and thereby alter protein measure. As frequencies of these protein-altering variants may vary between populations, they may drive differences in assay performance across populations. Here, we propose the first examination of protein measure agreement in diverse populations, using newly generated affinity-based proteomics data across four large studies of participants within the Trans-Omics for Precision Medicine (TOPMed) consortium. After quantifying the correlation of protein measures for each platform within each study, and the heterogeneity of these correlations between ancestral populations, I will identify technical and biological predictors of platform correlation. Next, I will analyze whole genome sequences for contributing individuals to identify genetic predictors of circulating protein abundances, as measured by each platform. I will characterize signals across platforms to identify protein-altering genetic variants which may drive assay interference, and synthesize these genetic associations with platform correlation coefficients to predict assay efficacy. Finally, I will model protein measures from each platform against relevant cardiometabolic phenotypes to evaluate agreement of epidemiological associations across platforms, and explore whether adjustment for protein-altering genetic variants in epidemiological models may improve the concordance of associations. The results of this analysis will disentangle reproducible protein measures and association results from those likely reflecting genetically- driven assay interference, within and across populations, and present new strategies to harmonize downstream associations for the impacted proteins across platforms.

NIH Research Projects · FY 2025 · 2025-09

The increasing frequency of weather extremes such as flooding on the incidence of malaria and other vector-borne diseases is an issue of substantial public health importance. Therefore, there is an urgent need to develop effective mitigation and control strategies. In May 2020, we piloted a post-flood malaria chemoprevention intervention in rural western Uganda. When compared to two neighboring villages where no intervention was deployed, we estimated a 53% reduction (aRR 0.47, 95% CI 0.34 – 0.62, p<.01) in malaria incidence in the six months post-intervention. Building on these results, the scientific aim of this proposal is to evaluate the effectiveness of a targeted, time-limited malaria chemoprevention intervention with and without larval source management (LSM) to reduce excess disease burden in a perennial, high-transmission setting following severe flooding. Our premise is that the intervention will reduce the parasite reservoir during the critical “rebound” period, when vectors are re-establishing habitats thereby maintaining relatively low levels of transmission until conditions have returned to the pre-flood baseline. Specifically, we will: Aim 1: Determine the effectiveness of chemoprevention with or without peri-domestic larvicide application to reduce the incidence of P. falciparum malaria after severe flooding. We will conduct a cluster randomized trial in 50 villages (144 clusters) in rural western Uganda, where flooding relatively predictable, to compare: (i) LLIN distribution (control), (ii) LLIN distribution plus three, monthly rounds of DP provided to children ≤12 years of age (DP), and (iii) LLIN+DP with three months of bi-weekly application of larvicide in the peri-domestic space (DP+Bti). Hypothesis: Chemoprevention will achieve a sustained (≥6 months) reduction in malaria incidence of ≥30% compared to the control arm without the emergence of artemisinin resistance. Aim 2: Elucidate the evolution of vector populations, feeding behaviors, and sporozoite rate in affected villages up to one year after flooding. We will perform weekly surveillance for adult and juvenile Anopheles mosquitoes in sentinel households and within the peri-domestic space. Collected mosquitoes will be counted, and identified to species and tested for the presence of P. falciparum sporozoites. Hypothesis: Flooding will create innumerable small pools favored by An. gambiae complex resulting in predominance of An. gambiae s.s. with potential for emergence of An. arabiensis as important vector in setting of widespread LLIN use.23 Aim 3: Evaluate the relative cost-effectiveness of chemoprevention (Aim 3A) and assess social and economic impacts on households (Aim 3B). We will conduct baseline and longitudinal quantitative surveys in sample clusters to evaluate impacts of the interventions on (i) economic activities and income, (ii) health seeking and expenditures, and (iii) mental health and well-being. These data will inform a cost effectiveness analysis of chemoprevention compared to the control (i.e., relative cost per case averted). Hypothesis: Chemoprevention will mitigate negative effects of malaria in the aftermath of a flood.

NIH Research Projects · FY 2026 · 2025-09

Assessment of Twin Gestation with AI-Assisted Ultrasound ABSTRACT This research aims to address the significant challenge of diagnosing and managing twin pregnancies in resource-constrained settings, where access to obstetric ultrasound services is often limited. Despite the clear benefit to both mothers and babies, many low- and middle-income countries and rural areas of the United States face substantial barriers to routine obstetric ultrasound, owing to the high costs of equipment and the need for trained sonographers to perform the scans. Consequently, twin pregnancies frequently go undetected until late in gestation, or even delivery, limiting the opportunities for effective intervention and management. In this proposal, we outline an innovative solution to twin assessment in resource-constrained settings. Point-of- care ultrasound (POCUS) devices offer a cost-effective alternative to traditional ultrasound machines. A "blind sweep" protocol enables providers without formal sonography training to conduct basic scans. And deep learning AI models analyze blind sweeps to make diagnoses. The overarching goal of this R01 proposal is to produce a suite of five essential tools that will allow obstetric providers working in resource-constrained settings to better diagnose and manage twin gestations. These tools include (a) diagnosis of twins, (b) estimating twin gestational age, (c) assessing chorionicity, (d) monitoring individual growth and growth concordance, and (e) establishing fetal presentation. The twin-specific AI models that underly tools a and b have already been developed and are ready to be tested in prospective diagnostic accuracy studies (Aims 1 and 2). Tools c, d, and e require additional data collection and model development (Aim 3). Our objectives will be supported by a common protocol in Chapel Hill, North Carolina, and Lusaka, Zambia, where our team has long-standing clinical research sites. We will enroll 700 women (350 twin and 350 singleton pregnancies) in a shared clinical cohort serving our diagnostic studies (Aims 1 and 2) and an additional 100 twin pregnancies (complementing 350 from our shared clinical cohort) to aid in new model development (Aim 3). The successful execution of our project holds significant potential for expanding care access to an exceedingly vulnerable population.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT During brain development, neurons undergo dramatic growth and branching to establish the intricate morphology required for neural network connectivity and function. This cellular expansion involves a substantial and rapid increase in neuronal surface area and therefore requires extensive insertion of material into the plasma membrane. Defects in neuronal morphogenesis can result in improper synaptic connectivity, neurodevelopmental disorders, and psychiatric syndromes. Insertion of membrane material is facilitated by SNARE-mediated exocytosis. Two SNARE proteins, Vesicle-associated membrane protein (VAMP) 2 and VAMP7, are enriched in the embryonic brain and are associated with distinct vesicle populations even prior to synapse formation. While knockout studies in mice have shown that both VAMP2 and VAMP7-mediated vesicle fusion are required for proper neuronal morphogenesis, the specific mechanisms regulating the trafficking and fusion of these vesicles during development is not known. My preliminary data reveal that VAMP2 and VAMP7 mediate non-synaptic exocytic events that cluster in different areas of the developing neuron, suggesting the existence of distinct regulatory pathways governing their distribution and fusion. Additionally, my preliminary data indicate that VAMP2-mediated exocytosis is sensitive to Ca2+ chelation, whereas VAMP7-mediated exocytosis is not, mirroring the differential Ca2+ sensitivity of VAMP2 and VAMP7 vesicle pools observed at the synapse of mature neurons. This suggests a potential role for Ca2+ signaling in the regulation of VAMP2 and VAMP7- mediated exocytosis during neuronal morphogenesis. Endoplasmic reticulum (ER)-PM membrane contact sites, key regulators of Ca2+ signaling, may also be involved in this regulatory process. The goal of this proposal is to investigate the regulation of VAMP2 and VAMP7-mediated exocytosis during neuronal morphogenesis using hypothesis driven science coupled with unbiased proteomic analysis. I will utilize total internal reflection fluorescence (TIRF) microscopy and advanced image analysis techniques to quantitatively define the interplay between SNARE-mediated exocytosis, Ca2+ signaling, and ER-PM membrane contact sites during neurite outgrowth in stage 2 developing neurons. Using a proximity biotinylation approach I will define the interactome of VAMP2 and VAMP7 during neuronal morphogenesis, providing insights into the molecular machinery regulating their trafficking and fusion. These findings will advance our understanding of the complex regulation of exocytic membrane trafficking during neuronal development. Successful completion of this proposed research will provide advanced training in live cell microscopy, the development of advanced image analysis pipelines, proteomics, and bioinformatics.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Mycobacterium tuberculosis (Mtb) is one of the most persistent pathogens that has been transmitted within the human population for millennia. The convention of this field has largely focused on understanding the mechanisms of the virulence genes that make Mtb pathogenic. However, the bacterial genetic basis underlying tuberculosis (TB) transmission—a critical aspect of TB epidemiology—remains understudied and poorly understood. This knowledge gap exists because transmission is a population-level phenotype and effective measurement tools have only recently become available. Whole-genome sequencing of Mtb strains from population-based cohorts has enabled the assessment of transmissibility of different Mtb strains. My research, alongside others, has shown that in various high-burden settings, the majority of ongoing TB transmissions are driven by a small subset of Mtb strains have evolved to be highly transmissible. The identification of highly transmissible strains and their phylogenetically neighboring low-transmissible strains suggests transmissibility of Mtb is an evolving trait, offering a new research opportunity to study the bacterial genetic basis underlying TB transmission. In this proposal, I plan to apply a novel research framework that integrates population genomics and cutting- edge experimental tools to investigate the bacterial genetic basis of high transmissibility. Specifically, (1) I aim to identify key mutations driving high transmissibility in Mtb by leveraging a global collection of 120,000 Mtb genomes from 72 countries. Using population genomics approaches, I seek to identify convergent mutations and common pathways evolved by highly transmissible strains from different countries. (2) I will develop a novel high-throughput bacterial physiological profiling system to characterize the altered bacterial functions in highly transmissible strains under various host-related conditions. (3) Building on my previous findings, I will investigate the role of redox mutations (sseA and trxB2), evolved by highly transmissible strains in Tibet and Peru, in promoting transmission by increasing resistance to reductive stress and facilitating granuloma necrosis. The primary goal is to identify common mechanisms evolved by highly transmissible strains in different countries to enhance transmission. If successful, these insights can guide the development of new preventive strategies tailored to halt the spread of these strains. My cross-disciplinary expertise in population genomics, high-throughput bacterial phenotyping, and classical bacterial genetics uniquely positions me to execute the planned research successfully. Additionally, the approaches developed in this project can be applied to investigate other important bacterial pathogens. This project will also provide a fully genotyped and phenotyped panel of highly transmissible strains as an open resource for other research groups to test new interventions, such as antibiotics and vaccines.

NIH Research Projects · FY 2025 · 2025-09

Aggressive B-cell lymphomas are increasingly common but underdiagnosed in people with HIV (PWH) in Malawi. In 2013, we established the Kamuzu Central Hospital (KCH) Lymphoma Study, a prospective observation cohort of pathologically confirmed lymphomas in Malawi that includes uniform diagnosis, standardof-care therapy, and five-year clinical and laboratory follow-up for participants. We have shown that treating lymphoma in PWH is safe and effective, and the Lymphoma Study has become a leading regional resource for clinical and translational research studies of HIV-associated lymphomas. However, current diagnostic strategies are prohibitively expensive which limits the reach of effective lymphoma care, and the pivotal genomic studies from our group are primarily being performed outside of Malawi. Herein, we propose longread nanopore sequencing approaches that require minimal capital investment, and limited lab infrastructure to molecularly diagnose and subclassify common aggressive B-cell lymphomas in PWH. The proposal builds on our work in HIV-associated lymphomas, cutting-edge expertise in long-read nanopore sequencing, and, most critically, and research infrastructure at the UNC-Project Malawi research site. We aim to develop effective, resource-appropriate diagnostic tools and subsequently investigate implementation of nanopore sequencing across a network of sites while performing ongoing clinical, computational, and economic assessments. This project builds capacity to perform advanced molecular characterization of HIV-associated lymphomas in Malawi and leverages ongoing clinical, research, and training programs.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Despite intensive prevention efforts from federal and state agencies, suicide remains a leading cause of mortality in the United States (US), increasing by 36% between 2000-2022. In 2023 alone, 49,303 Americans lost their lives to suicide. The impact of suicide is particularly high among vulnerable populations such as adolescents and formerly incarcerated individuals. Although much research has focused on causes and predictors of death from suicide, progress in suicide prevention has been hampered by data linkage and methodological challenges. While national mortality rates are known, entities with the potential to implement large suicide prevention initiatives – insurers, health systems, and departments of corrections – lack the linked data to monitor suicide mortality in their populations, establish benchmarks, and create an evaluation framework for prevention efforts. Many individuals who die from suicide have had recent contact with a health insurance, health care, or correctional system, representing critical missed opportunities to implement suicide prevention measures. Funded from 2020-2025, our original “INSPIRE” grant (R01MH124752) established a living suicide mortality surveillance system in North Carolina (NC) by securing all necessary data approvals and completing individual-level linkages between state death records and large datasets on four key populations covering 10-22 years depending on the data source: the publicly insured (NC Medicaid), privately insured (large single private insurer), a large integrated health system (UNC Health), and releasees from the Department of Adult Corrections. We have updated these linkages annually and have used this surveillance system to define benchmarks of suicide mortality in each population; evaluate policy impacts; identify risk factors for suicide mortality, attempts, self-harm, and suicidal ideation; and predict short and long term risk of suicide outcomes . Having invested the expertise, resources, and time to establish the INSPIRE living surveillance system and demonstrate its potential, RFA-MH-25-135 now provides an opportunity to untap the full potential of this surveillance infrastructure and further advance suicide prevention efforts. Over the next five years, we will enhance the surveillance system by continuing annual updates, including additional important populations (older adults), and performing enhanced cross-population linkages. We will utilize the established system to evaluate the impact of several recent suicide prevention efforts in NC and build risk assessment tools tailored to specific sub-populations and suicide mortality subtypes. Finally, we will move this research beyond Big Data as we engage end users (clinicians and health administrators) to plan implementation of the risk assessment tools into health care settings where they can be best utilized to connect people in mental health crisis with the help they need to prevent suicide-related outcomes.

NIH Research Projects · FY 2026 · 2025-09

Project Summary Tengfei Li, Ph.D, is a statistician committed to developing advanced statistical methods to better understand the human brain function and structure, the aging process, cognition and brain disorders using multi-modal neuroimaging and genetic data. The research he proposes, entitled “Tracking brain structural trajectories for early detection and prognosis of Alzheimer's Disease”, will unify multi-center multi-modal datasets using cutting-edge statistical and machine learning techniques. The result will be a comprehensive set of life-span brain structural reference standards and a centile-based explainable artificial intelligence (AI) system, which facilitate timely and precise detection of brain structural changes in the early stages of Alzheimer's Disease (AD) and AD related dementias (ADRD) and aid in the diagnosis and prognosis of the early stages of AD. Candidate: Dr. Li is a Research Assistant Professor in the department of Radiology, University of North Carolina at Chapel Hill. He completed his Ph. D in probability and mathematical statistics prior to his postdoctoral fellowship in biostatistics at the University of Texas MD Anderson Cancer Center. Dr. Li's previous work, which has been published in high-impact journals such as Science and Nature Genetics, focuses on imaging-genetic analysis and big data integration. The proposed career development plan will build upon his previous training with four training goals to enhance his trajectory toward becoming an independent investigator: 1) Build a solid understanding of the neurodevelopmental basis of AD/ADRD; 2) Develop novel methodologies and strengthen in-depth understanding of AI in neuroimaging and AD/ ADRD analyses; 3) Develop professional skills to analyze multicenter big data; and 4) Establish leadership, expand collaborative networks, and translate diagnostic tools into clinical practice. Dr. Li and his primary mentor, Hongtu Zhu, Ph.D, have assembled a strong team of co-mentors to guide Dr. Li through the proposed training and research activities. Research: Despite an increased availability of neuroimaging datasets, the analysis of brain structure change by AD is limited by narrow age ranges, small sample sizes, and a lack of critical covariates. Besides, there is no reference standard against which to anchor individual differences in brain morphology from multisite images that may or may not align with growth charts. Dr. Li proposes to overcome these limitations by establishing life-span brain structural developmental reference standards that can be used to identify abnormal brain aging/developmental patterns in the early AD phases, including cortical/subcortical structures and white matter tracts. In Aim 1, he will develop a Functional Harmonization Regression Modeling (FHRM) framework to integrate multi-site data; in Aim 2, he will build a brain Local Growth Curve Modeling (LGCM) system to build the normal brain structural aging trajectories and track their variations associated with early phases or high-risk AD groups; and in Aim 3, he will develop a computationally efficient, easily interpretable and state-of-the-art Centile-based Explainable AI (CEAI) system in delineating the preclinical AD, MCI and AD transitions. This award will provide Dr. Li with the training and research needed to be successful for future, high- impact multi-center multimodal studies and behavioral/cognitive performance related to brain aging and AD. This will significantly contribute to the field and support Dr. Li's transition towards scientific independence.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Antibiotic resistance is a growing global crisis exacerbated by a dearth of novel antibiotic classes and the rapid emergence of resistance to novel antibiotics of existing classes. In the United States, nearly 1 in 7 deaths attributed to bacterial pathogens are caused by Enterobacter or Klebsiella pneumoniae, members of the order Enterobacterales with among the highest rates of cephalosporin-and carbapenem-resistance. One form of antibiotic resistance is heteroresistance, a type of phenotypic heterogeneity in which an isolate harbor a minority resistant subpopulation within a majority susceptible population. The resistant subpopulation is dynamic: unlike other resistant subpopulations which include persisted and tolerant cells, the heteroresistant subpopulation grows during antibiotic exposure and becomes the majority population as the susceptible cells are killed but will return to baseline frequency when antibiotic exposure ends. Thus, this transient, low frequency resistant subpopulation is challenging to detect clinically and may be a cause of unexplained treatment failure. Experiments aimed to understand the mechanistic basis for heteroresistance to the novel cephalosporin cefiderocol revealed that many Gram-negative isolates harbor resistant subpopulations that encode many copies of an extended-spectrum beta-lactamase gene. The primary focus of preliminary experiments, Enterobacter isolate RS, encodes the blaSHV-5 beta-lactamase in a region flanked by identical sequences. This region undergoes homology-mediated gene amplification, creating the resistant subpopulation. While the subpopulation is resistant to cefiderocol and other cephalosporins because of the multiple copies of blaSHV-5, the underlying principles which result in multiple copies are unknown. To characterize these principles of amplification, experiments proposed here will in Aim 1 define the factors governing homology-mediated amplification in the Enterobacter chromosome, focused on understanding how the size and content of the amplified region, and the homologous sequences, influence gene amplification and subpopulation dynamics in vitro and during murine models of antibiotic therapy. Additional experiments identified that the activity of the SHV-5 enzyme dictates the extent of amplification. Chemical or genetic inhibition of SHV-5 activity resulted in greater gene copy number in the resistant subpopulation, establishing a connection between enzyme activity and subpopulation dynamics. Thus, experiments in Aim 2 will determine the effects of beta- lactamase activity on gene amplification and subpopulation dynamics in Enterobacter and K. pneumoniae in vitro and in vivo. Findings from this work will describe the factors governing gene amplification and provide insight into diverse forms of phenotypic heterogeneity that result from gene amplification, as well as enhance our understanding of the dynamics between resistant subpopulations and antibiotic treatment.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Emergency first responders are at significant risk for injury, illness, and death during disaster response and recovery operations, particularly with the growing frequency and severity of disaster events such as Hurricane Helene that impacted Western North Carolina in the Fall of 2024. This disaster continues to draw first responders from different areas of the state and regionally, presenting a unique opportunity to assess environmental hazards’ impact on first responders. The National Institute for Environmental Health Sciences’ (NIEHS) Disaster Research Response Program’s (DR2) crosscutting strategy to address the consequences of disasters notes that critical opportunities like this are often missed to better understand physical and mental health impacts of these events. It is clear that we must determine if first responders are able to recognize and be fully prepared for the unique hazards they may encounter. An important gap exists in our understanding of current and emerging safety, health, and psychosocial hazards placing first responders at risk, particularly as they occur in areas where the complexity of response is increased (e.g., landslide risk). A critical need exists to study first responders who have recently participated in response and recovery operations to best determine hazards and how personal, organizational, and other factors of the job place them at risk. The objective of this project is to better identify the range of environmental hazards encountered by first responders during response and recovery operations, and how these hazards impact perception of risk, particularly psychosocial hazards, and then identify key areas to improve training for responder preparedness. The proposed study is innovative due to: (1) the use of the National Institute for Occupational Safety and Health's Burden, Needs, and Impact method, a well-validated tool that aids identification of environmental impacts and research priorities; (2) a Total Worker Health® (TWH) approach to assess environmental factors impacting responder well-being; and (3) the extraordinary opportunity to aid first responders per the time sensitive nature of the event. In Aim 1, we will conduct one-on-one interviews and focus groups with a variety of first responders (e.g., firefighters, EMS, law enforcement) to identify the current and emerging personal hazards encountered in the response which increase their risk of injury, illness, and mental distress. In Aim2, we will apply the TWH framework to inform responder preparedness through identification of broader workplace and organizational factors impacting responder safety, health, and well-being. In Aim 3, we utilize a participatory approach working with first responders to develop training recommendations to better protect their safety, health, and well-being. The expected outputs and outcomes of this project will inform the NIEHS DR2 goal of “identifying both immediate and long-term potential physical and mental health effects of disaster events” utilizing insights obtained directly from first responders actively engaged in the Hurricane Helene response.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Antibiotic treatment failure imposes a significant global health and economic burden. Even in the absence of resistance mutations, the recurrence of infections can be attributed to a resilient subset of genetically identical bacteria. This phenomenon, recognized as antibiotic-tolerant persister formation, also facilitates the development of resistance, underscoring a critical challenge that demands attention. Mounting evidence suggests that the intracellular environment plays a pivotal role in the formation of persister cells, serving as a crucial reservoir for these bacterial populations. Among the most difficult-to-treat bacterial pathogens, Staphylococcus aureus can survive antibiotic exposure within host cells, including macrophages, and subsequently resume growth when conditions become favorable again. However, the intracellular niche of S. aureus in vivo and the underlying mechanisms driving persister formation remain largely enigmatic. This knowledge gap can be attributed to the challenge of distinguishing viable persisters from non-viable bacteria and the low frequency of cells harboring live bacteria (less than 0.01% of overall cells extracted from mouse organs). To address this hurdle, I have developed inducible reporter strains that will enable the identification of intracellular reservoirs in vivo. Using single-cell approaches, I will identify the host cells containing S. aureus and the antibiotic-tolerant persisters (Aim 1.1). Both the host and intracellular bacterial transcriptomes will be probed using single-cell RNA-sequencing (scRNA-seq) and a novel single-cell bacterial RNA-seq technology called proBac-seq (Aim 1.2 & 1.3). I have also established a high-throughput screening platform aimed at identifying energy modulators of intracellular S. aureus, as the low energy state is the central characteristic underlying persister cells’ insensitivity to antibiotics. My preliminary screen of >4,700 drug-like compounds has identified a compound (KL1) that increased the energy level of intracellular S. aureus as well as its susceptibility to antibiotics. Further understanding of its mechanism of action using the chemoproteomic method thermal proteome profiling (TPP) will unravel novel therapeutic target(s) for devising antibiotic adjuvants (Aim 2.1). Lastly, I intend to explore potential host-directed energy modulators from a select library of established mammalian kinase inhibitors targeting intracellular immune responses (Aim 2.2). These studies will provide a comprehensive understanding of the interplay between host responses and antibiotic tolerance within the intracellular milieu.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY Psychological trauma is often unavoidable and an established contributor to persistent neuropsychiatric sequelae (PNS). Yet only a proportion of trauma-exposed persons are vulnerable to PNS, and the mechanisms underlying this vulnerability are largely unknown. Trauma and related PNS have been associated with advanced biological aging, measured with both molecular markers and brain imaging. Among molecular markers, epigenomic markers (so-called “epigenetic aging”) combine DNA methylation (DNAm) at multiple age-related genomic sites to predict disease outcomes. Complementarily, telomere shortening is another molecular marker independently associated with aging and disease. The overall objective of this application is to determine the extent to which molecular aging markers right after trauma and their changes over time predict and co-occur with PNS trajectories and age-related brain alterations. The central hypothesis is that advanced molecular age contributes to high risk for PNS and concomitant age-related alterations in brain function and structure after trauma. The rationale for this application is that combining integrative assessments of molecular age with longitudinal deep phenotyping and brain imaging data after trauma can yield novel predictors and insights to guide interventions for promoting brain health and alleviating PNS across the age spectrum. The central hypothesis will be tested with three specific aims: 1) Determine the extent to which integrative molecular aging markers at the time of trauma predict PNS; 2) Uncover the temporal and genomic site-specific dynamics of molecular age along PNS trajectories; 3) Define the functional and structural brain alterations dynamically associated with molecular age after trauma. To this end, the application leverages established multi-ancestry discovery and replication cohorts of women and men presenting to the emergency department (ED) right after psychological trauma exposure. Participants have provided whole blood DNA at ED presentation and then were followed with deep neuropsychiatric phenotyping, functional and structural brain imaging, and additional DNA sampling over the ensuing 12 months. Leveraging this unique study design and already available DNA in both cohorts, integrative molecular age will be longitudinally determined by measuring epigenetic aging and telomere length. Moreover, longitudinal whole methylome sequencing (WMS) data will be generated in a cohort subset to develop unbiased, novel epigenomic markers of aging in association with PNS. Lastly, key findings will be generalized to non-ED settings using larger-scale DNAm data available in neuropsychiatric cases and trauma- exposed controls from diverse military and civilian cohorts of the Psychiatric Genomics Consortium. This research is innovative as it will assess molecular age at the time of trauma as a predictor of PNS, interrogate the entire methylome with WMS, and integrate longitudinal molecular and phenotypic measures with brain imaging. This work is significant as it can establish molecular age measured at ED presentation and other settings as a novel predictor and potential treatment candidate for promoting brain health and alleviating trauma-related PNS.

NIH Research Projects · FY 2025 · 2025-08

The 29th United States Conference on HIV/AIDS (USCHA) 2025, hosted by NMAC in collaboration with the University of North Carolina (UNC) Center for AIDS Research (CFAR), will focus on “Aging with HIV: Challenges and Opportunities for People Over 50”. Advances in HIV treatment have greatly increased the life expectancy of persons with HIV. Recent data suggests that of approximately 1.1 million people with HIV in the United States over 53% were aged 50 and older. This age demographic shift presents unique challenges and opportunities requiring focused attention and tailored strategies for intervention. The specific aims of the conference are: 1) To advance scientific knowledge on the intersection of aging and HIV. 2) To enhance clinical practice and care for older adults living with HIV. 3) To promote policy and advocacy efforts that address the needs of older adults with HIV. 4) To foster community engagement and support networks for older adults living with HIV. NMAC has a 30 year history of successfully drawing healthcare providers, state and local health department leaders, researchers, and people with HIV to participate in the annual USCHA. The average attendance at USCHA between 2022-24 was 3,000 persons. Given the critical and timely need to address the needs of people with HIV over 50, we anticipate similar or slightly higher attendance for the 2025 USCHA.