Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2024-07

Zebrafish have greatly advanced toxicology and environmental studies as a valuable animal model because they are easy to manipulate, breed, and observe during development. However, the absence of universal standards significantly impedes scientific progress and therefore human and environmental health. Standard ontologies and data models are acutely needed to improve data compatibility within and across species and domains. We propose to address these needs by creating community-built standards for annotating zebrafish toxicological exposure and their phenotypic outcomes (toxicophenotypes). We will design and deploy a toxicophenotype data model that will advance ontologies such as the Zebrafish Phenotype Ontology (ZP). This data model will allow integration and interoperability across species and across toxicological studies. We will create a toxicophenotype annotation toolkit that will allow users to annotate their data conforming to the newly created toxicophenotype data model and ZP, and therefore create “born-interoperable” data. Finally, we will instantiate a zebrafish toxicophenotype atlas web application. This atlas will serve as a visual definition of the standards and their documentation for examining variations of specific phenotypes by laboratories in the community. Users will be able to explore and query exposures and phenotypes of interest and see example images demonstrating the phenotypes. This project will community-governed by diverse stakeholders in toxicology and environmental health sciences to ensure fit-for-purpose design and sustainability. Towards that end, we will create robust, transparent structures for deciding on standards content, structure, and versioning; attribution policies; and content management and access practices. We will coordinate requirements gathering, documenting best practices for using the standards and annotations, usability testing, and plans for sustainability. Realizing interoperable toxicophenotypic data is crucial to improve data integration across scale and granularity; thereby accelerating an understanding of environmental influences on health.

NIH Research Projects · FY 2025 · 2024-07

Calprotectin is the most abundant protein in neutrophils. Our surprising discovery that calprotectin can bind and hydrolyze multiple β-lactam antibiotics forms the basis of this proposal. In particular, calprotectin hydrolyzes and inactivates the penicillin sub-class that includes ampicillin, amoxicillin and oxacillin, which are among our most important and most widely used antibiotics. The identification of calprotectin as the first known host protein with β-lactamase activity has major implications for the treatment of infection. Indeed, our preliminary data demonstrate that human neutrophils hydrolyze β-lactam antibiotics in tissue culture and that oxacillin treatment is more effective in calprotectin-deficient mice than in wild-type mice. In Aim 1, we will determine the impact of calprotectin’s β-lactamase activity on antibiotic efficacy in vivo through three sub-Aims. We will first assess the ability of calprotectin to hydrolyze β-lactams during E. coli and S. aureus infection in mice. We will determine how the β-lactamase activity of calprotectin alters antibiotic- mediated growth inhibition and killing of pathogens during bacteremia and wound infection. We have shown that the serine β-lactamase inhibitor, sulbactam, blocks the β-lactamase activity of calprotectin. Therefore, we will next examine the capacity of various approved β-lactamase inhibitors to inhibit the β-lactamase activity of calprotectin and determine the capacity of inhibitors to improve β-lactam efficacy in mice, against penicillin sensitive bacteria, lacking any native β-lactamase production. Finally, we will assess whether β-lactam binding or inhibition of β-lactamase activity of calprotectin affects the innate immune functions of calprotectin, including metal sequestration and TLR4 activation. In Aim 2, we will elucidate the molecular mechanism(s) of calprotectin’s β-lactamase activity through two sub-Aims. Our preliminary evidence suggest calprotectin has multiple β-lactam binding sites and binds β- lactams with a high affinity. We will first fully characterize the β-lactamase activity of calprotectin, including dependence on parameters such as metal ions, salts, pH, temperature, and multimeric state. We will test hypotheses concerning reaction mechanism through enzymatic characterization of site-directed mutants. We will complement kinetic methods by determining the co-crystal structure of calprotectin bound to various inhibitors of β-lactamase activity. The completion of our Specific Aims will comprehensively characterize the newly discovered β- lactamase activity of calprotectin, determine its contribution to antibiotic efficacy in vivo and identify new avenues for drug development. Our proposal is significant due to potential impact on multiple aspects of treatments for bacterial infections. Our proposal is conceptually innovative in establishing a new paradigm for how calprotectin affects the outcome of bacterial infection.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT In sub-Saharan Africa, universal access to antiretroviral therapy (ART) for pregnant and breastfeeding persons, known as Option B+, has rapidly expanded ART access in low and middle-income countries, yielding declines in vertical HIV transmission and improvements in viral suppression. However, disengagement from perinatal HIV care in the perinatal period has emerged as a major challenge to sustaining the success of Option B+. Interventions to support sustained engagement in perinatal HIV care are urgently needed. Perinatal depression (PND) is common among WLHIV and is an important barrier to HIV care engagement. PND is defined as onset of depression in pregnancy and/or within the first 3-6 months postpartum. Among WLHIV in sub-Saharan Africa, 30-40% experience prenatal depression and 20%-30% experience postnatal depression. PND in WLHIV has been linked to increased HIV viral load, postpartum disengagement from HIV care, and reduced infant HIV testing, making PND an important barrier to reaching UNAIDS 2025 goals and ending the HIV epidemic. To address these gaps, NIMH (NOT-MH-21-270) has prioritized testing interventions to address PND among WLHIV in the perinatal period, including their impact on HIV-related outcomes among the mothers and their infants. However, few evidence-based interventions exist that address both perinatal mental health and HIV care outcomes for WLHIV, and even fewer are positioned for rapid translation into practice through incorporation of a rigorous implementation science framework at the effectiveness trial stage. Funded by NIMH (R34MH116806), our team enhanced The Friendship Bench, an evidence-based counseling intervention, to address PND and HIV care engagement for WLHIV. In a pilot individually randomized controlled trial, the Enhanced Friendship Bench demonstrated high levels of acceptability, feasibility, and fidelity to the intervention protocol, as well as preliminary efficacy to improve both PND and engagement in HIV care for WLHIV in Malawi. Building on this work, the objective of this proposal is to evaluate the efficacy of the Enhanced Friendship Bench, compared to usual care, in a fully powered randomized controlled trial to improve PND, HIV care engagement, and secondary infant health outcomes through 12 months postpartum; test hypothesized mediators and moderators that will help elucidate mechanisms of intervention action; and collect key implementation science metrics in a Hybrid Type I design to accelerate translation of findings into practice. Our team is exceptionally well qualified to address these aims given our development and enhancement of the Friendship Bench for perinatal WLHIV; our expertise in HIV, perinatal mental health, and implementation science; and our longstanding research infrastructure in Malawi. Completion of the proposed aims will shift clinical practice by providing a culturally appropriate, scalable counseling intervention that improves PND and engagement in HIV care outcomes for WLHIV that is poised for rapid translation into practice.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT The National Lung Screening Trial demonstrated the efficacy of lung cancer screening (LCS) with low-dose computed tomography in reducing 5-year lung cancer mortality by 20%, albeit with high rates of false-positive results. The US Preventive Services Task Force recommends annual LCS with LDCT for individuals ages 50- 80 years who currently or formerly (quit within 15 years) smoked with a 20-pack-year history. Despite the task force Grade B recommendation, a major concern of LCS is the potential harm associated with false-positive results which are defined as having one or more nodules requiring follow-up and no change or lung cancer diagnosis after one year. Specifically, false positives may burden the patient, clinician, and healthcare system due to the economic and psychological effects of extra imaging and invasive procedures, fear of missing cancer, and complications from procedures for benign lesions. There is a need to better estimate the burden and the factors that affect false positive results in real-world practice over multiple rounds of screening to inform patient-clinician conversations, guide the development of interventions to reduce false positive rates, and optimize the risk-benefit ratio of LCS. The primary objective of this proposal is to identify the burden of false-positive results at the patient, clinician, and facility levels and to understand the impact of false-positives across the screening continuum. Using rich sources of longitudinal LCS data for >40K LCS exams from 17 facilities linked with state-based cancer registry data, we will accomplish the objectives through the following specific aims (1) Identify attitudes, knowledge, and experiences with false-positive results from LCS among patients and clinicians; (2) Determine patient-, exam-, radiologist-, and facility-level factors that affect false-positive findings on lung cancer screening; (3) Assess the cumulative risk of a false-positive LCS exam over 5- and 10- rounds of annual screening across multilevel factors. An estimated 14.5 million US adults are eligible for LCS and are at risk of experiencing a false positive result. This study will generate in-depth knowledge about the potential harms of false-positive LCS results. These findings may inform patient-clinician shared decision-making conversations or may help to reduce false positive rates by identifying modifiable radiologist or facility-level factors.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY Rigidity is an important property of molecules that increases the likelihood of their success as drugs. Our goal with support of this MIRA grant is to develop reactions that enable molecular rigidification. We describe a research program that enables access to rigid small and medium sized-strained rings. Our innovative approach to obtaining rigid molecules is to use sulfone anions as carbene surrogates. We will also develop new methods to stabilize highly reactive alkylidene units using palladium catalysis. The knowledge gained from these studies using reactive carbenes will also be used to enable skeletal editing of heterocycles. We will apply the reactions developed in this proposal to synthesize complex natural products and their derivatives with tubulin binding properties and antifungal properties.

NIH Research Projects · FY 2025 · 2024-07

Multifaceted Characterization of Early Human Brain Development Abstract In the first few years of life, the human brain develops dynamically in both structure and function. Many neuro- developmental disorders are associated with aberrations from normative growth during this critical period of brain development. The longitudinal high-resolution MRI data of children from birth to 5 years of age, made available through the Baby Connectome Project (BCP), affords unprecedented opportunities for precise charting of early brain developmental trajectories in order to understand normative and aberrant growth. Dedicated computational tools have been developed at the University of North Carolina at Chapel Hill for accurate processing and anal- ysis of baby MR images, which typically exhibit dynamic heterogeneous changes across time. The goal of this secondary analysis project is to apply these tools to the data acquired via the BCP to investigate structural and functional connectomes, tissue macrostructure and microstructure, and their interplay during early brain devel- opment. In Aim 1, we will investigate the hierarchical organization of the cerebral cortex by analyzing areal differences in neuroanatomical characteristics involving cortical morphology and microstructure. We will utilize our infant-centric pipeline to delineate cortical geometry by constructing white matter and pial surfaces, based on which macroscopic features of cortical morphology, such as thickness and curvature, and microscopic features of myeloarchitecture and cytoarchitecture, such as neurite and soma densities, will be extracted and analyzed. For completeness, tissue macroscopic and microscopic measurements of subcortical structures will also be included for investigation. In Aim 2, we will study brain development in terms of dense vertex-wise cortical connectivity. We will use our infant-centric diffusion model and tractography algorithm to significantly improve the delineation of white matter pathways, particularly in superficial white matter with characteristically low diffusion anisotropy, and to reduce gyral bias in establishing dense connectivity of cortical surface vertices. Vertex-wise stationary and dynamic functional connectivity will also be analyzed. In Aim 3, we will investigate the interplay of multiple developmental traits during the first years of postnatal brain development. We will study brain subnetworks in association with motor, language, and visual development. We will assess the associations of these networks with psychological assessments such as the Mullen Scales of Early Learning (MSEL) with subdomains including gross/fine motor, receptive/expressive language, and visual reception. This project will involve the utilization of multimodal MRI, including structural, diffusion, and functional MRI, to provide a more complete picture of human brain development. Successful completion of this project will empower the neuroscience community with improved understanding of the development of structure and function of the human brain during its infancy.

NIH Research Projects · FY 2025 · 2024-07

Project Summary / Abstract Virtually fatal 60 years ago, pediatric cancers are now curable for 85% of children who have access to contemporary treatments and robust supportive care. However, for the 80% of children born in low and middle- income countries (LMIC), the picture is less promising. When viewed from a global perspective, only 30% of children in LMIC will survive their diagnosis of cancer, highlighting the stark contrast between high-income countries (HIC) and LMIC and representing one of the largest disparities in global health. While curable, cancers in children are not preventable. With no opportunities for prevention, but cost-effective malignancy-specific treatments increasingly available, an accurate diagnosis often represents the difference between potential survival and certain death. Modern methods to ascertain a correct diagnosis, such as flow- cytometry, cytogenetics, and molecular genomics, are standard of care in HIC, but unavailable for many LMIC treatment centers. Transformative diagnostic approaches utilizing cost-effective, scalable technology are urgently needed. As genomic sequencing platforms become more accessible and analytical capabilities expand, integrating sequencing into childhood cancer diagnostics represents an opportunity to overcome the global “pathology gap.” Classification of pediatric cancers using short-read RNA sequencing is established in research settings, but the capital and maintenance expenses of this approach are prohibitive for most LMIC. However, nanopore sequencing platforms require minimal capital investment and limited lab infrastructure, providing an opportunity for cost-effective, scalable molecular diagnostic technology in pediatric cancer treatment centers. We propose to establish and validate robust classification models for pediatric leukemias, extracranial solid tumors, and lymphomas using nanopore sequencing. We will leverage the close partnership between investigators at the University of North Carolina at Chapel Hill, St. Jude Children’s Research Hospital, and Indus Hospital and Healthcare Network in Pakistan to optimize technical and analytics methodologies for nanopore sequencing of challenging samples, specifically formalin-fixed, paraffin embedded specimens. We will harden existing protocols for deployment in LMIC and incorporate our analytic pipeline into integrated nanopore sequencing and base-calling hardware as a step toward affordable point-of-care cancer diagnostics for end users. Our objective is to develop and validate a cost-effective, sustainable, adaptable solution for the clinically relevant diagnosis and core genomic classification for childhood cancers in LMIC using a globally accessible platform.

NIH Research Projects · FY 2026 · 2024-07

Multifaceted Characterization of Early Human Brain Development Abstract In the first few years of life, the human brain develops dynamically in both structure and function. Many neuro- developmental disorders are associated with aberrations from normative growth during this critical period of brain development. The longitudinal high-resolution MRI data of children from birth to 5 years of age, made available through the Baby Connectome Project (BCP), affords unprecedented opportunities for precise charting of early brain developmental trajectories in order to understand normative and aberrant growth. Dedicated computational tools have been developed at the University of North Carolina at Chapel Hill for accurate processing and anal- ysis of baby MR images, which typically exhibit dynamic heterogeneous changes across time. The goal of this secondary analysis project is to apply these tools to the data acquired via the BCP to investigate structural and functional connectomes, tissue macrostructure and microstructure, and their interplay during early brain devel- opment. In Aim 1, we will investigate the hierarchical organization of the cerebral cortex by analyzing areal differences in neuroanatomical characteristics involving cortical morphology and microstructure. We will utilize our infant-centric pipeline to delineate cortical geometry by constructing white matter and pial surfaces, based on which macroscopic features of cortical morphology, such as thickness and curvature, and microscopic features of myeloarchitecture and cytoarchitecture, such as neurite and soma densities, will be extracted and analyzed. For completeness, tissue macroscopic and microscopic measurements of subcortical structures will also be included for investigation. In Aim 2, we will study brain development in terms of dense vertex-wise cortical connectivity. We will use our infant-centric diffusion model and tractography algorithm to significantly improve the delineation of white matter pathways, particularly in superficial white matter with characteristically low diffusion anisotropy, and to reduce gyral bias in establishing dense connectivity of cortical surface vertices. Vertex-wise stationary and dynamic functional connectivity will also be analyzed. In Aim 3, we will investigate the interplay of multiple developmental traits during the first years of postnatal brain development. We will study brain subnetworks in association with motor, language, and visual development. We will assess the associations of these networks with psychological assessments such as the Mullen Scales of Early Learning (MSEL) with subdomains including gross/fine motor, receptive/expressive language, and visual reception. This project will involve the utilization of multimodal MRI, including structural, diffusion, and functional MRI, to provide a more complete picture of human brain development. Successful completion of this project will empower the neuroscience community with improved understanding of the development of structure and function of the human brain during its infancy.

NIH Research Projects · FY 2025 · 2024-07

Project Summary This exploratory grant seeks to evaluate linkages between social reward processing in autism spectrum disorder (ASD) and systemic inflammation. The social motivation theory of autism posits that reduced motivation to interact with people and decreased pleasure derived from social interactions may derail typical social development and contribute to the emergence of social communication deficits in ASD. This framework highlights the centrality of impaired brain reward circuitry functioning to the etiology of ASD and suggests that when young children with ASD lack the motivation to participate in activities where social skills are typically forged, the resulting impoverished social environment contributes to the emergence of social communication impairments in the disorder. There is increasing evidence that inflammatory processes contribute to ASD risk and pathogenesis, and that neuroinflammation, in turn, interferes with social reward processing. However, no research to date has examined relations between immune function and neural responses to social rewards in ASD. The objective of this proposal is to investigate relations between (i) neural responses to social rewards, measured via electroencephalography (EEG), (ii) a blood-derived composite marker of systemic inflammation, and (iii) ASD symptoms and quality of life. Additionally, half of participants will be recruited from a companion study collecting positron emission tomography (PET) data using a translocator protein 18 kDa (TSPO) tracer to measure neuroinflammation centrally, and an exploratory goal of this proposal is to investigate relations between EEG-based responses to social rewards, PET-derived (i.e., central) measures of neuroinflammation, and a blood-derived (i.e., peripheral) measure of systemic inflammation from the portion of autistic participants who completed TSPO PET scans. This project will provide a deeper understanding of relations between neural responses to social rewards, immune functioning, and symptom expression in ASD. It also has the potential to contribute to the development of EEG-based measures that may be suitable endpoints in future mechanistic trials investigating novel ASD interventions targeting inflammatory processes.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY - ABSTRACT The overall goal of this proposal is to characterize Factor XII (FXII) functions in sickle cell disease (SCD) and their contribution to venous thrombosis (VT), vaso-occlusive crisis (VOC) and end-organ injury. SCD is a hematologic disorder caused by a single nucleotide mutation of the β-globin gene. Hypoxia-induced sickling of red blood cells (RBCs) results in painful recurrent VOC, hemolytic anemia and cumulatively, in multi-organ damage. A chronic hypercoagulable state with increased risk of venous thrombosis contributes to increased morbidity and mortality among patients with SCD. New targeted anticoagulant therapies are still associated with increased rates of bleeding. We previously demonstrated that FXII contributes to the development of VT through both its enzymatic (FXIIa) functions as well as the thromboinflammatory effects of zymogen FXII-mediated activation of urokinase plasminogen activator receptor (uPAR) on neutrophils. Recently, we found that patients with SCD exhibit chronically enhanced FXII activation compared to healthy controls. Moreover, we showed that FXII expression is elevated in SCD neutrophils, and that FXII contributes to neutrophil activation and adhesion in SCD. Importantly, treatment with an anti-FXII antibody that blocks both zymogen and enzymatic functions of FXII(a) significantly attenuated experimental VT and microvascular stasis in a murine model of SCD. These findings support the central hypothesis that FXII contributes to vaso-occlusion and VT in SCD through distinct cellular and molecular mechanisms. Using clinical samples and well-established murine model of SCD together with novel pharmacologic and genetic approaches, our goals in this application are to: i) identify disease-specific triggers of FXII activation; ii) characterize FXII effects on neutrophil functions; iii) investigate the relative contributions of different cellular sources, and compare the zymogen versus protease functions of FXII in vascular stasis and VT; iv) determine the effect of long-term FXII deficiency on disease progression and early mortality associated with SCD. Our goal is to delineate the mechanisms by which FXII and its downstream effectors drive SCD pathologies, which will lay the foundation for future therapeutic approaches to inhibit FXII.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Necrotizing enterocolitis (NEC) is a devastating, rapidly progressive gastrointestinal disease of prematurity that affects ~10% of neonates born at < 1500 grams. The risk of NEC is highest for the most preterm neonates, and one infant dies from NEC in the United States each day. The mortality rate for NEC approaches 50% if surgical resection of necrotic bowel is required. Intestinal injury in NEC results from a dysregulated cycle of unrestrained inflammation and intestinal epithelial cell damage. Despite four decades of research, outcomes have not improved for neonates with NEC due to critical knowledge gaps in our understanding of mechanisms underlying disease pathogenesis. The studies outlined in this proposal will characterize new signaling mechanisms regulating intestinal epithelial injury in NEC, determine how these pathways are modulated by microbial metabolites, and use this knowledge to identify new dietary strategies or therapeutic targets. Our laboratory found that activation of the aryl hydrocarbon receptor (AhR) by the dietary ligand indole-3-carbinol (I3C) downregulates the expression of inflammatory cytokines and reduces intestinal injury during experimental NEC. We will now advance our understanding of the anti-inflammatory signaling pathways in the neonatal intestine by examining a pathway closely related to AhR characterized by activation of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2). In adult murine colitis models, Nrf2 agonists attenuate intestinal injury. Similarly, our preliminary data reveal that dietary Nrf2 agonist administration reduces inflammation in our murine model of NEC and our unique and innovative microfluidic model incorporating patient-derived intestinal epithelial cells and microbiome. Based on our preliminary data and the published literature, we hypothesize that intestinal epithelial injury will be (1) exaggerated in the absence of Nrf2 signaling in intestinal epithelial cells, (2) downregulated by Nrf2 agonist production by intestinal microbes, and (3) significantly attenuated by therapeutic agents activating the Nrf2 pathway. For these studies, we will utilize specialized genetically modified strains of mice, a humanized NEC model, and our microfluidic model incorporating human neonatal intestinal epithelium. The K08 candidate is a neonatologist-scientist who has transitioned her research focus to intestinal epithelial cell biology and host-pathogen interactions in the developing intestinal tract. Her mentor, Dr. Misty Good, is an international expert in this field and is completely dedicated to the candidate’s success. The extensive training plan outlined in his proposal will facilitate the achievement of her goal of leading an R01- funded laboratory dedicated to improving the health of vulnerable neonates through research focused on intestinal biology and the pathophysiology of NEC.

NIH Research Projects · FY 2026 · 2024-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Many medications are modified by intestinal bacteria, positioning them as an emerging and signif- icant driver of overall drug response (efficacy and toxicity) beyond host factors. Considering the importance of bacterial drug metabolism during preclinical development will uncover drug-induced toxicities in relevant tissues, prompt lead retooling and reduce adverse events in human trials. My career goal is to develop pharmacomicrobiomics, the emerging study of drug-microbiota interac- tions that drive variable drug responses. My research goal is to develop a mechanistic under- standing of how bacterial enzymes mediate toxicity and adverse effects of drugs, and to devise strategies to reduce prevent, or treat microbe-mediated drug toxicities in organs involved in drug disposition. Over the next five years, the MIRA will support my efforts to utilize the bacterial en- zyme β-glucuronidase (GUS) as a tool to elucidate mechanisms by which bacterial drug metabo- lism alters host physiology. GUSs hydrolyze and reactivate glucuronide-conjugated drugs formed by Phase II metabolism of host UGT enzymes. Deconjugated aglycones are active and can be toxic to tissues involved in drug disposition, e.g. the intestines. Molecular and structural features driving GUS’ substrate preferences are known, genetic manipulation of many GUS encoding bac- teria is feasible, and selective GUS inhibitors are in preclinical development. Combining this foundational postdoctoral work with high-throughput primary cell culture platforms, I propose two interrelated research programs: Program 1 will test the hypothesis that gut microbiota contribute to interindividual variability in bacterial reactivation of drug-glucuronide conjugates. Program 2 will test the hypothesis that drug disposition-associated tissues derived from different people vary in their susceptibility to bacterial drug metabolism. This proposal is the first systematic analysis of bacterial metabolism of host Phase II conjugates, and their contribution to drug-induced gut inju- ry. This foundational work will facilitate addition of bacterial drug metabolism into future physiolo- gy-based pharmacokinetic modeling to aid accurate prediction of drug side effects and efficacy. The proposed research is significant as it will systematically interrogate the contribution of bac- terial drug metabolism to interindividual variability in drug responses. It will develop phenotypic biomarkers of bacterial drug reactivation and resultant drug-induced injury and serve as a primer for future pharmacomicrobiomic studies to improve precision medicine.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Anhedonia (ANH), the reduced ability to anticipate or experience pleasure, is a major transdiagnostic contributor to disability, inferior quality of life, and a core characteristic of multiple severe psychopathologies. ANH emerges in adolescence prior to the onset of these disorders and is highly prevalent (up to 20%) in youth 13-15 years. However, ANH in adolescence is poorly understood and interventions remain few. A growing body of evidence suggests that stress exacerbates ANH and dysregulated acute stress response mechanisms may critically contribute to ANH. Yet, little is known about the role of stress neurobiology in the emergence, course, and severity of ANH in adolescence, a critical developmental period marked by significant pubertal changes and steep maturation of neural circuits implicated in the recalibration of the stress-response systems and mood regulation. The objective of the current proposal is to characterize how biological acute stress response mechanisms influence the severity and trajectory of ANH. ANH research to date has almost exclusively focused on single-system physiological responses to acute stress, such as autonomic nervous system (ANS - parasympathetic and sympathetic nervous system branches - PNS-SNS) or endocrine (HPA - Hypothalamic Pituitary Adrenal) system responses. New evidence supports the need for multi-system approaches to detect clinically meaningful patterns of stress responses associated with ANH. Recent research from our labs (PI Belger and Co-PI Roubinov) has shown that (1) discrete physiological latent stress- response profiles (Buffered, Vigilant, Sensitive) that integrate unique patterns of responses across HPA and ANS systems have additional utility in predicting both clinical and functional development outcomes beyond single-system biomarkers. In this proposal, we will examine how a novel constellation of complex neurobiological stress response mechanisms interactively contributes to ANH in adolescence. Using a multimodal approach including functional MRI (Magnetic Resonance Imaging), physiological HPA and ANS stress reactivity, and clinical and cognitive assessments, the proposed study will be the first to integrate physiological and neural acute stress response mechanisms to derive multisystem stress response biotypes (MSRB) and explore their contributions to ANH severity and trajectory in adolescence. We will study 192 adolescents ages 13-15 years experiencing a range of ANH at baseline, 10 months, and 20 months across 2 aims. In Aim 1 we will identify multisystem stress response biotypes (MSRBs) integrating physiological and neural responses to acute stress in adolescents and examine their longitudinal stability. In Aim 2, we will determine the characteristics of multisystem latent stress profiles that predict the severity and trajectory of ANH in adolescence. Across both aims, we will examine age, sex, and pubertal stage as covariates. IMPACT: This project will contribute a novel framework for understanding and modeling the complex multisystemic biological mechanisms governing stress responsivity during adolescence. Understanding the individual and interactive contributions of physiological and neural systems to ANH presents a unique opportunity to develop specific interventions that target malleable stress systems in adolescents with ANH.

NIH Research Projects · FY 2025 · 2024-07

Mental health problems are a leading cause of learning and behavioral difficulties among children worldwide and in Tanzania where research has documented high rates of youth depression, anxiety, and suicide, 43% of the population is under the age of 15, and access to treatment services is limited. One theoretically important driver of youth mental health is the primary school environment. Schools have the potential to foster cognitive, emotional, and social functioning that, in turn, promotes mental wellbeing. Further, school experiences are thought to play a critical role in shaping the coping systems that support positive adjustment in youth who have experienced early life stressors. As such, intervening to improve school determinants holds promise as a potentially effective “upstream” prevention approach. Our understanding of what to target for intervention is limited, however, because has almost no rigorous longitudinal research has been conducted to identify specific malleable determinants of the school environment that shape mental health. Our long-term goal is to develop practical, scalable set of strategies that leverage positive aspects of schools to promote the mental health of youth in Tanzania and in other resource-limited settings. Specific aims of the study are to: (1) develop a contextually meaningful set of measurement tools to comprehensively assess the primary school environment in Tanzania; (2) examine how different dimensions of the school environment work to impact mental health trajectories of Tanzanian youth; and (3) determine how features of the school environment can be leveraged to buffer the impacts of exposure to early life stressors (violence exposure and deprivation). To accomplish these aims, we will collect data from teachers, caregivers, and students in 60 randomly selected primary schools in the Mwanza region of Tanzania. For aim 1, we will develop a robust set of observational and survey measures of the school environment, using qualitative research to investigate contextual relevance and a psychometric study to examine the validity and reliability of the adapted measures. To accomplish Aims 2 and 3, a random sample of 50 5th grade students in each of the 60 selected schools (total n=3,000) will be surveyed at 6-month intervals over 3 years for a total of 6 assessment waves. School observations and caregiver and teacher surveys will enable multi-informant assessment of key constructs. Longitudinal mediation models will assess the influence of school environment factors on mental health and examine emotional, cognitive, and social processes as explanatory mechanisms (Aim 2). Moderated mediation models will determine whether the negative mental health impacts of stressors are buffered for children in supportive school environments. Qualitative interviews with parent/child dyads will provide contextualized information about these processes (Aim 3). We expect the study will have a positive impact by identifying malleable factors in the school environment that can be targeted by whole school interventions to promote youth mental health in resource-limited settings.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT The success of PD-1-targeted immune checkpoint inhibitor (ICI) therapy has revolutionized cancer treatment, becoming a new standard of care for various types of cancer. However, the treatment outcomes for triple negative breast cancer are still limited. To enhance the efficacy of checkpoint inhibitor therapy, it is important to gain a deeper understanding of PD-1 regulation in the tumor microenvironment. B cells in the tumor immune microenvironment have been understudied, despite their growing recognition as key players in shaping the tumor microenvironment. The presence of tumor-infiltrated B cells indicates the prognosis of many types of cancer. Interestingly, our previous research in pre-clinical models has shown that the sensitivity to dual ICI therapy, including the PD-1-targeted treatment, is dependent on B cells, and to a lesser extent, CD8+ T cells. These findings emphasize the importance of studying the PD-1 signaling in B cells to improve the clinical outcomes of checkpoint inhibitor therapy. The goal of this study is to understand how PD-1 signaling pathway regulates B cells in the tumor microenvironment. One of the main mechanisms for PD-1 to modulate T cell function is through reprogramming T cell metabolism, allowing them to meet the energy requirements needed for differentiation into various subpopulations. We hypothesize that the PD-1 activation on tumor-infiltrated B cells alters the metabolic program and B cell fate decision similar to T cells. PD-1-targeted therapy potentially impacts B cells in proliferation, survival, differentiation, and function through metabolic reprogramming, which serve as a main mechanism for PD-1 to regulate the antitumor function of B cells. Aim 1. To evaluate our hypothesis, we will dissect how PD- 1-targeted therapy modulates B cell differentiation through metabolic switch ex vivo. We will combine primary murine B cell culture with state-of-the-art metabolic profiling tools to determine if and how PD-1 blockage directly altered B cell differentiation and metabolism. This approach allows the evaluation of the glycolysis and fatty acid metabolic machinery in B cell to control these cells differentiation into germinal center B cells, antibody-secreting cells, and other functional subsets of B cells. Aim 2. We will use PD-1 fate mapping mice (PD- 1Cre/tdTomato.Rosa26floxp-YFP) and transgenic breast cancer models to examine how PD-1-blockage guides B cell fate decisions and function in vivo. Elucidating the interaction between B cells and the PD-1 signaling pathway, which has so far escaped the attention of the cancer immunology field, would strongly enhance the clinical activity of ICI therapy in tumors where B cells are critical to the anti-tumor response.

NIH Research Projects · FY 2026 · 2024-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Alternative splicing is an RNA processing mechanism that explains how single genes can produce more than one transcript. Alternative splicing is a rule rather than an exception: in humans, more than 90% of genes undergo alternative splicing, consistent with the increased cellular and functional complexity of higher eukaryotes. Genome wide studies keep identifying multiple splice variants for thousands of genes but how they differentially or similarly function in the cells is an arena that only few groups get into. The splicing field is heavily focused on how alternative splicing is regulated for example by RBPs, transcription, or epigenetics components. In contrast, how alternative splicing impacts protein function and physiology is a much less investigated area. After our current NIGMS R01 ends, this MIRA/R35 will greatly help us keep building our research program in this niche. Our lab is interested on how alternative splicing influences key cellular process such as membrane trafficking, cytoskeleton dynamics, transcription, local translation, and phase separation in large, highly specialized cells such as cardiomyocytes and myofibers. We are curious of how this interplay impacts organ physiology, and how similar or different this is in other type of large, highly specialized cells such as neurons that exert very different roles in our bodies. Our MIRA proposal will tackle this broad interest from three Angles, that are independent from each other but at the same time will synergize our discoveries. Angle 1 will study how RNA processing regulates cytoskeleton dynamics in skeletal muscle cells which are also highly mechanosensitive. This is significant because muscle diseases caused by aberrant RNA processing show intracellular architecture and mechanical defects. Angle 2 will examine the contribution of alternative splicing to phase separation and local translation. This is significant because skeletal muscle cells are syncytial tubes with numerous nuclei that need to coordinate transcription within nuclei and translation in their shared cytoplasm. Angle 3 will define the role of splicing on regulating transcription. This is significant because transcription factors and chromatin regulators control thousands of downstream programs that in turn drive cellular differentiation, cell fate, and tissue function. Overall, our MIRA research will build mechanistic and physiological models of how RNA processing drives organ development and tissue identity acquisition and maintenance.

NIH Research Projects · FY 2025 · 2024-07

Cancer is a significant cause of morbidity and mortality in the United States and across the world. Approximately 2 million new cases are diagnosed annually, and about 600,000 people die from the disease in any given year. The Cancer Undergraduate Research Education Program (CUREP) aims to address cancer by expanding the pool of future cancer researchers within the biomedical workforce, with a long-term goal of enhancing cancer prevention, treatment, and clinical outcomes. We propose to achieve this by recruiting outstanding undergraduate students pursuing a STEM major, such as Biology, Biochemistry, Chemistry, Life Sciences, Public Health, Nursing, and other biomedical sciences, and providing them with hands-on research experience, career development, and professional coaching through a summer undergraduate research program in basic science and public health cancer research. The program has three specific aims: 1) Provide undergraduate students with hands-on experience in cancer research during a ten-week summer internship. 2) Increase student interest in pursuing careers in cancer research by offering professional development activities essential for gaining admission to graduate and professional schools. 3) Provide year-long group coaching (covering summer, fall, and spring) to help students integrate academic and life skills and increase their chances of successfully completing their undergraduate degrees. CUREP offers participants opportunities to expand their knowledge, explore various aspects of cancer research, and develop the skills needed for successful careers in cancer research. The program's key components include direct research experience under the guidance of principal investigators conducting cancer research. These faculty mentors possess expertise in basic cancer biology, population science, translational cancer research, and cancer health outcomes. Our program's structure builds upon previous experiences in undergraduate summer programs. Our innovative approach integrates academic study with life skills coaching, creating a comprehensive program to prepare young scientists for future challenges. Students will conduct independent research under the supervision of PI mentors, graduate students, and postdocs in laboratories or research teams specializing in population science. Their research experiences will be complemented by training in ethics and various professional and career development activities, culminating in research presentations at the end of the summer. A distinguishing feature of CUREP is its yearlong coaching component, encompassing study skills, academic writing, time management, self-efficacy, career planning, and wellness. This holistic approach enables participants to integrate academic and social/life skills effectively. Ultimately, CUREP aims to increase the participation of undergraduate students pursuing careers in cancer research, thereby enhancing the biomedical workforce.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY Despite unprecedented clinical success of chimeric antigen receptor (CAR)-T cell therapy against tumors, widespread application is limited by lengthy and labor-intensive ex vivo manufacturing procedures that result in: (i) very high costs of therapy of up to half of a million dollars; (ii) delays of weeks or months to infuse CAR-T cells to patients with rapidly progressing disease; and (iii) heterogeneous composition and terminal differentiation of infused CAR-T cells as a result of ex vivo culture that limit CAR-T cell engraftment and persistence. Effort to overcome these limitations have focused on closed and automatic manufacturing devices to contain the labor needed to manufacture CAR-T cells ex vivo, and allogeneic off-the-shelf CAR-T cells have been proposed to overcome the need of CAR-T cell manufacturing for each single patient. Despite significant achievements in this space, reducing the time, costs and regulatory burden remains a deep unmet need in CAR-T cell therapy and significant reducing or eliminating ex vivo procedures remains a critical unmet need. In vivo generation of CAR-T cells would eliminate the need for ex vivo procedures, prevent the terminal differentiation of ex vivo expanded CAR-T cells and ensure the potency and longevity of autologous T cells as compared to allogeneic CAR-T cell products that are extensively manipulated to prevent rejection and graft- versus-host disease The research outlined in this proposal develops new biomaterials approaches to reduce the time and effort to produce CAR-T cells in vitro, to enhance CAR-T cell efficacy and persistence in vivo and, finally, to eliminate ex vivo manipulation entirely by generating CAR-T cells entirely within the patient. We propose that biomaterial scaffolds displaying anti-CD3/CD28 antibodies and releasing pro-proliferative interleukins will mediate simultaneous activation and viral transduction of T cells without centrifugation (spinoculation) or transduction agents (retronectin, polybrene) and will facilitate ex vivo genetic reprogramming of T cells by reducing the time and expense of activating naive T-cells and transducing them with viral vectors. We next propose that directly implanting scaffolds seeded with peripheral blood mononuclear cells and CAR- encoding viral vectors will promote release of CAR-T cells into circulation, eliminating ex vivo CAR-T isolation and proliferation protocols to promote a less differentiated cell phenotype associated with longer in vivo persistence. Finally, we propose that, through the inclusion of encapsulated T-cell attracting cytokines, implanted biomaterial scaffolds will generate CAR-T cells entirely in situ through recruitment of host T cells to the scaffold, in-scaffold reprogramming of recruited T cells with resident CAR-encoding viral vectors, and release of reprogrammed CAR-T cells. We expect that our results will provide a basis for a general cellular therapeutic strategy and promote widespread patient access. In addition to the obvious applications in blood cancers, this rational materials-based approach for cellular manufacturing will be adopted to program therapeutic lymphocytes in solid tumors and for other diseases.

NIH Research Projects · FY 2025 · 2024-07

SUMMARY ABSTRACT Clostridioides difficile is among the most common causes of nosocomial infections with disease ranging from antibiotic-associated diarrhea to pseudomembranous colitis. Secreted toxins are largely responsible for disease development, yet many aspects of C. difficile physiology and virulence remain poorly understood. Recent work has revealed that C. difficile produces two colony morphotypes—a rough colony variant and a smooth colony variant—and can reversibly switch them. This phenomenon is conserved among diverse C. difficile strains. The two morphological variants differ in several ways. Bacteria from rough colonies are longer and often found in chains, exhibit greater surface motility and diminished swimming motility, produce less biofilm biomass, and show greater pathogenicity in animal models compared to the smooth colony counterpart. Our prior work linked colony morphology and the correlated phenotypes to the expression of genes encoding a signal transduction system, CmrRST, but the molecular mechanisms by which C. difficile develops rough and smooth colony morphologies have yet to be defined. The objective of this study is to identify genes required for formation of each colony morphotype and to determine the roles of these genes in cell morphology, motility, biofilm formation, and virulence. In Aim 1, we propose three complementary yet independent mutagenesis and screening strategies to identify genes required for rough and smooth colony development. This work takes advantage of mutants designed to yield only one colony morphotype. The in vitro phenotypes of the mutants obtained through the genetic screens will be evaluated to determine the broader impact of the identified genes on C. difficile physiology and virulence traits. In Aim 2, we will examine how one identified mutation results in strictly rough colonies and assess its ability to colonize and cause disease in a mouse model of C. difficile infection. Similar strategies will be used to characterize additional mutants obtained in Aim 1. Pilot studies with the proposed approaches have identified multiple candidate genes predicted to affect cell division. As such, these gene products may serve as new targets for therapeutic development, yet almost none of these genes have been previously studied. The proposed research will identify factors contributing to disease relevant phenotypes including cell division proteins, facilitating efforts to combat C. difficile infection.

NIH Research Projects · FY 2026 · 2024-07

Deciphering the Stepwise Regulatory Mechanisms of Genome Folding In multicellular organisms, the precise regulation of gene expression across the different cell types and developmental stages is achieved through the orchestration of cis-regulatory elements, notably enhancers. These elements are positioned distantly from gene promoters and exert their function by establishing physical interactions with them. Crucially, these interactions depend on the three-dimensional folding of the genome. Any disruptions to this highly orchestrated genome folding, which can give rise to dysregulated gene expression, have been implicated in the pathogenesis of a myriad of diseases, including cancer and developmental disorders. Hence, unraveling the mechanisms that govern genome folding becomes a paramount endeavor in order to gain a comprehensive understanding of the precise control of gene expression, both in healthy physiological contexts and in the aberrant states associated with disease. Over the last two decades, significant advancements in genomic studies employing proximity-ligation and sequencing techniques have revealed the cohesin complex as a key driver of genome folding. Recent groundbreaking single-molecule imaging experiments have demonstrated cohesin's motor activity, directly engaging in loop extrusion to fold linear DNA in vitro. However, it has been largely elusive how cohesin-mediated genome folding is regulated in the living cells. To answer this question, we will focus on three research topics: 1. How do nuclear bodies influence cohesin function? 2. What are the post-translational modifications (PTMs) on cohesin-associated factors and their impact on genome folding? 3. What are the cytosolic signals that regulate genome folding through cohesin and how do they do so? Cohesin-mediated genome folding is achieved through highly dynamic processes of cohesin loading/loop extrusion, extrusion stalling, and cohesin unloading. We will dissect and pinpoint the specific process within the genome folding that are regulated by nuclear bodies, PTMs and cytosolic signals. Collectively, our proposed studies aim to unveil novel molecular mechanisms governing genome folding, providing a comprehensive understanding of how genome folding is regulated in living cells and its contribution to disease states.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Male breast cancer is critically understudied due to both the rarity of the disease and the exclusion of men from breast cancer trials leading to a significant knowledge gap about how male breast tumors respond to therapy. The efficacy of existing endocrine therapies in men with ER+/HER2- breast cancer have never been compared prospectively, and this is the goal of the ongoing ETHAN trial. In this trial, 60 patients are randomized to 3 endocrine regimens for a 3-week window phase, followed by biopsy, then a 4-month neoadjuvant phase +/- the CDK4/6 inhibitor abemaciclib, followed by surgical resection. The trial is well powered to compare differences in response rates, measured in the window phase by a 50% reduction in Ki-67, and in the neoadjuvant phase by residual cancer burden index. This design creates an unprecedented opportunity for translational studies using samples from pre and post treatment timepoints annotated with clinically relevant response endpoints. The goal of this proposal is to leverage patient samples from the ETHAN trial to understand and predict how male breast tumors respond to therapy. We hypothesize that multigene response signatures will predict response to ET in men with ER+ breast tumors. To test this hypothesis, in Aim1 we will determine endocrine therapy response signatures using whole transcriptome sequencing of pre and post treatment specimens. We will determine how each drug treatment results in changes in gene expression and compare these to female predictors and response signatures. We will perform stratified analysis of responders and non-responders to discover biomarkers which will be validated using large cohorts of patients with ER+/HER2- breast cancer treated with endocrine therapy. In Aim2, we will utilize our single cell processing pipeline which we used to show distinct responses in malignant cell subpopulations within female ER+/HER2- breast tumors, to dissect heterogeneity in endocrine therapy response within male breast tumors. This analysis will define features of low abundance cell populations that may be driving the resistance phenotype. Finally, in Aim3, we will address a critical lack of experimental models of male breast cancer by using our organoid infrastructure to generate male patient derived organoids. We will use organoids to test estrogen receptor gene regulatory function and assess the preclinical efficacy of targeted therapeutic strategies. Cumulatively, these studies will address a significant knowledge gap of why therapies are effective or i neffective in men with breast cancer. These findings will generate predictors of response, identify pathways driving resistance, and generate and utilize experimental models to uncover the intrinsic biology of male breast tumors. If successful, these insights will lead to more precise treatment regimens and improve outcomes for men with breast cancer.

NIH Research Projects · FY 2024 · 2024-07

ABSTRACT Adolescent dating violence (ADV) is highly prevalent among girls in the US and can have devastating consequences for mental health, including increased risk for depressive symptoms and suicidal ideation. As such, there is a critical need for effective depression and suicide prevention strategies for ADV girl survivors. Restorative justice (RJ) is one policy-based justice and accountability response to ADV that has the potential to improve youth mental health disparities among ADV girl survivors. RJ allows survivors to narrate their ADV stories and offer perpetrators an opportunity to acknowledge and remedy the harm, and engage in rehabilitation, which supports survivor preferences and fosters healing. To our knowledge, no study has examined whether state-level RJ policies and their implementation impact depressive symptoms or suicidal ideation among ADV girl survivors over time and whether impacts differ by race or ethnicity. The proposed fellowship builds on the applicant's prior background to prepare her for a career as an independent mixed methods researcher and behavioral scientist with expertise in structural- and community-level prevention intervention efforts for gender-based violence and its mental health consequences. The specific fellowship goals are to: 1) deepen health policy methods and advanced multilevel and causal quantitative analysis skills, 2) develop and strengthen intersectional mixed methods skills in public health, and 3) refine academic writing and communication skills. These goals will be accomplished through coursework and workshops, mentorship from the sponsor team, and execution of the proposed research project including dissemination of study findings. The specific aims of this proposed research are to: 1) examine associations between state RJ policies with depressive symptoms and suicidal ideation among ADV girl survivors, and test the effect modification of race/ethnicity (N=82,400), 2) among states with a RJ policy, examine associations between policy implementation with depressive symptoms and suicidal ideation among ADV girl survivors, and test effect modification of race/ethnicity (N=41,200), and 3) qualitatively describe perceptions of mental health and preferences for justice and accountability among ADV girl survivors (N=30). The proposed research uses a sequential explanatory mixed methods (QUANT -> QUAL) approach in a two-part research project. Aims 1 and 2 leverage 4 waves of population-representative data in 48 states from the 2013, 2015, 2017, and 2019 waves of the Youth Risk Behavior Surveillance System (YRBS) and data on RJ policies specific to adolescents from a RJ legislative database. Aim 3 uses semi-structured interviews conducted with 10 white, 10 Black and 10 Hispanic ADV girl survivors to contextualize quantitative findings and identify ways to improve and tailor the justice response for ADV girl survivors. This proposal aligns with NIMH's youth mental health disparities call by examining how state policies impact mental health outcomes using longitudinal data, which has the potential to inform the mental health equity and enhancement of justice practices for ADV girl survivors.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY & ABSTRACT Abstract As the most common malignancy of the urinary tract among men and women, bladder cancer is estimated to surpass 80,000 new cases and 17,000 deaths is 2023. Accounting for most bladder cancers cases, urothelial carcinoma has a dismal survival rate of approximately 5% in the metastatic setting. APOBEC3A (A3A) and APOBEC3B (A3B) are members of a family of cytidine deaminase enzymes that catalyze the removal of an amino group from cytosine nucleotides generating a uracil in its place that serve as a source of mutations. A3A and A3B enzymes are commonly overexpressed in numerous cancer types, including urothelial carcinoma, with the APOBEC mutation signature seen in most cases of this cancer type. Due to their mutagenic activity, A3A and A3B have been implicated in altering the genomic landscape of urothelial carcinoma tumors over time. These alterations have the potential to augment tumor cell clonality that can lead to an enrichment of cells that are refractory to treatment and drive disease progression. Currently, it is unclear if both A3A and A3B drive APOBEC-induced mutagenesis, or if one enzyme plays a larger mutagenic role than the other. This proposal seeks the address the hypothesis that A3A and A3B enzymes differ in mutagenic activity leading to differences in promotion of intratumoral heterogeneity and response to anti-PD-1 immune checkpoint blockade inhibition. To address this hypothesis, two mouse bladder cancer cell lines entitled BBN963 and BBN976, that are representative of human muscle invasive bladder cancer, will be developed to express either HA-epitope tagged A3A or A3B, in a doxycycline inducible manner. Utilization of this model will allow for a direct comparison of A3A and A3B induced mutagenic activity, tumor cell clonality diversification, and modulation of response to treatment. Experiments detailed in Aim 1 will elucidate the driver of the APOBEC mutational signatures in urothelial carcinoma using in-vitro assays to analyze the predicted target sequences of APOBEC3 enzymes. Subsequently, Aim 2 will use DNA barcoding technology to compare the promotion of intratumoral heterogeneity by A3A and A3B. Lastly, Aim 3 will determine the influence of A3A and A3B on treatment response to anti-PD-1 therapy and the role these enzymes play in altering the tumor-immune microenvironment utilizing an in-vivo approach. Completion of the work outlined in these aims will result in the identification of the driver of APOBEC3 mutagenesis in urothelial carcinoma and highlight a potential target for inhibition for combination therapy with immune checkpoint blockade in patients with elevated APOBEC3 mutagenic activity.

NIH Research Projects · FY 2025 · 2024-07

The primary mission of the UNC MSTP is to train an outstanding group of students committed to becoming physician scientists, capable of bridging the gap between science and clinical medicine. We strive to achieve this goal by recruiting candidates with a variety of academic and research interests. We project a steady state of 96 students (12 new students per year) whose academic and extracurricular experiences are stellar, including substantial research experience and proven commitment to service. We plan to appoint MSTP trainees to the T32 during their first two years in the program. We will also appoint students returning to medical school after completing their PhD who do not hold F30 awards. Through our UNC MSTP, students will be able to pursue their graduate training in 15 individual departments and 5 curricula representing the Schools of Medicine, Public Health, Pharmacy, and the College of Arts and Sciences. Our leadership team includes Dr. Toni Darville and Dr. Mohanish Deshmukh, who serve as Co-Directors, Dr. Gaorav Gupta, who is the Associate Director, and Alison Regan, who is the Assistant Director. Our program emphasizes strong education in clinical medicine that is well integrated with superb research opportunities. Throughout their training, we promote a framework that compels students to define clinical implications of their research, and how research affects clinical care. Our program leverages resources from the UNC Clinical Translational Science Award (CTSA), the Biological and Biomedical Sciences Program (BBSP), links students with physician scientist role models, and structurally integrates research and clinical work in the thesis process, and in research-in-progress and clinical case conferences. During the PhD phase of training, students are engaged in a longitudinal clinical clerkship to maintain clinical skills. Graduating PhD students participate in the Extra Practice to Transition to Application Phase course to promote clinical confidence upon returning to medical school. These mechanisms help ensure that our students learn essential skills including conducting rigorous and reproducible research, oral presentation, and manuscript writing. Critical grant writing skills are taught through a rigorous F30 bootcamp and mock review sessions. Additionally, we meet extensively with each student throughout their training to ensure that they have a complete support system for research, clinical, and career mentorship via peer colleagues and faculty. Finally, we expose them to leadership and career development opportunities to prepare them for careers as Clinician Scientist leaders. Training outcomes include receiving a high percentage of honors for clinical rotations, obtaining first author publications, successfully competing for a variety of awards and independent funding (e.g., F30 awards from the NIH), completing the dual degree program with the average time of 8.1 years, matching at outstanding academic medical centers for residency, and continuing on to have impactful careers as physician scientists.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT In SSA, deaths occur in 5-10% of PLWH during the first year after initiating antiretroviral therapy (ART) and causes for these early deaths are not well elucidated in most patients. Many patients present with lymphadenopathy and are empirically treated for tuberculosis (TB), even when TB testing is negative, and may have undiagnosed lymphoproliferative disorders (LPDs). Access to lymph node biopsy and high-quality pathologic evaluation are limited and often lead to missed or delayed diagnosis which may contribute to early preventable deaths after ART initiation. The barriers to pathologic diagnosis of lymphadenopathy in Malawi have not been extensively studied. LPDs, including lymphomas and multicentric Castleman disease (MCD) are a common causes of lymphadenopathy in PLWH, but are likely delayed and/or underdiagnosed. In the KCH Lymphoma Study, we have enrolled all patients with newly diagnosed LPDs since 2013; 123/245 (50%) PLWH with lymphoma and 19/35 (54%) with MCD are from Lilongwe district which makes up only 10% of the catchment area. Given this marked overrepresentation of participants from Lilongwe, we hypothesize that there many undiagnosed LPDs, though a gap exists in understanding the burden of LPDs in this population. Critically, when diagnosed appropriately, LPDs are treatable and curable in SSA. To overcome barriers to diagnosis, innovative and rapid diagnostic technologies are urgently needed to improve the speed and accuracy of LPD diagnoses at the point of care. There a number of methods for EBV and KSHV measurement including traditional PCR and a point of care device known as TINY. TINY is a loop-mediated isothermal amplification (LAMP)-based device that allows amplification and quantification of DNA at point-of- care with operability even in settings where electricity is not available. Our collaborators have shown that TINY is sensitive and specific for diagnosing Kaposi sarcoma from skin biopsies and can potentially diagnose Burkitt lymphoma using EBV measurement though its utility at point of care for LPDs has not been studied. This proposal addresses current gaps in knowledge by 1) assessing multifactorial causes of diagnostic delay for LPDs in Malawi, 2) comprehensively describing clinical characteristics and diagnoses in PLWH presenting with lymphadenopathy from HIV clinics, and 3) assessing utility of EBV and KSHV measurement from FNA and blood for expedited diagnosis using standard PCR as well as TINY. In summary, this proposal will open a research focus that is distinct from my previous work as it will be my first time studying diagnostics and implementation science and the first time working in peripheral health centers. This multidimensional proposal will build towards my long-term goal of improving outcomes for PLWH and LPDs by providing critical information about prevalence of undiagnosed LPDs, key implementation challenges toward accurate and timely LPD diagnosis, and innovative diagnostic methods to overcome these challenges.