Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2026 · 2023-12

Abstract: Proposed is the development of readily available tools for radiochemistry based on tetrazine ligation— a bioorthogonal reaction with rapid kinetics that has been studied for over a decade as a uniquely powerful tool for radiochemical labeling of biomolecules. Limitations in accessing tetrazine and trans-cyclooctene (TCO) labeling precursors with appropriate kinetic and physicochemical properties have prevented the broader application of this technology. This proposal builds on several technological innovations in reaction chemistry that have been developed in the groups of PIs, including a methodology for introducing 18F into aromatic molecules via photocatalysis via C-H activation or aromatic substitution, a new methodology for the practical and scalable synthesis of trans-cyclooctenes with favorable kinetic and physiochemical properties, and new methodologies for the preparation of functionalized tetrazines. Through this collaborative technology proposal, our groups will leverage these methodologies for the development of new tools for the efficient radiolabeling of proteins in site- selective fashion and the application of these tools in vitro. In Aim 1, we will develop improved methods for the synthesis of 18F-labeled trans-cyclooctenes with balanced reactivity, hydrophilicity and stability. Late-stage photoisomerization will also be used to obtain highly reactive 18F-TCOs, and we will develop methodology for the site-selective attachment to the C-terminus of cancer-targeting proteins. In Aim 2, a rapid and mild method for the preparation of 18F-labeled tetrazine reporters will be developed using photoredox radiofluorination reactions for PET probe construction. In Aim 3, we will validate the newly developed 18F-TCOs and 18F-tetrazines using the hydrophilic FAPI ligand, the hydrophobic SR142948A, and proteins. The brain permeability of the 18F- TCO/tetrazine reporters will also be evaluated in normal rats. Because this application focuses solely on the development of 18F-labeling technologies, the studies in Aim 3 are primarily intended to provide important feedback to Aims 1-2, which will result in the design of 18F-TCOs and 18F-tetrazines of high utility to the broader radiolabeling community (in other words, comprehensive evaluation/characterization of specific probes will not be performed in this application). In summary, we are developing a general labeling toolbox to generate 18F labeled PET agents. We aim to develop labeling precursors that can be readily commercialized, so that the developed technologies will be widely used as efficient and simple labeling methods to modify biologically active small molecules/peptides/proteins/drugs, which could have a significant impact on medical imaging, drug discovery and development.

NIH Research Projects · FY 2025 · 2023-12

Abstract Individuals born prematurely are at increased risk for many chronic conditions, and respiratory disease represents a significant portion of the morbidity and mortality in this population. Better understanding of the etiology of chronic respiratory disease among those born preterm has the potential to improve diagnosis, treatment, and prevention. Asthma occurs more frequently and severely among those born preterm. The etiology of asthma is known to be multifactorial, involving innate inflammatory tendencies, potentiated by prenatal and early life exposures. Asthma exhibits differences in presentation, and pathophysiology, based on biologic sex, a phenomenon known as sexual dimorphism. Epigenetics represents a powerful tool for understanding early-life origins of chronic disease. Research from our group in the Extremely Low Gestational Age Newborn (ELGAN) cohort has illustrated the critical role of epigenetic and gene expression changes in the placenta as they relate to later development of a host of chronic conditions. As a temporary organ during gestation, the placenta forms a barrier between the developing fetus, the mother, and by extension, the outside world. It is a critical conduit through which the effects of environmental exposures are filtered. The placenta also plays critical roles in directing fetal growth and development, and establishing the neonatal immune system, while also protecting the fetus from rejection by the maternal immune response. The placenta is also highly sexually dimorphic, responding differently to prenatal exposures, and conferring risk differently for chronic disease, based on the biologic sex of the infant. Prior work from other groups has shown epigenetic changes in immune-related genes from cord blood cells in association with the later development of childhood asthma among children born at term. Preliminary analyses from EGLAN have shown differential epigenetic changes to immune-related genes in the placenta that are associated with asthma development in a largely sex-dependent manner. No other published studies have examined placental epigenetic factors as antecedents of asthma. We propose to further examine placental epigenetic mechanism and gene expression and their association with development of asthma at age 10 years. Our study would be the first to provide a comprehensive examination of epigenetic mechanisms and gene expression in the placenta, as well as epigenetic changes in neonatal blood on day of life 1 as they relate to later development of asthma among ELGANs. The examination of the role of differences based on biologic sex is also novel. These investigations will elucidate important general and sex-specific genetic pathways that form the basis of the developmental origins of asthma. Enhanced understanding of these will allow for improved diagnosis, treatment, and perhaps even prevention of asthma in a particularly vulnerable population.

NIH Research Projects · FY 2025 · 2023-12

Pertussis is a re-emerging public health concern, and its resurgence is correlated with a switch from whole-cell to acellular vaccines, which prevent disease, but not colonization or transmission of the causative agent Bordetella pertussis. In order to limit the mortality and morbidity associated with pertussis, particularly in infants, a combination of more effective vaccines and antibiotic therapies are needed. Their development requires a better understanding of the molecular basis for Bordetella pertussis pathogenicity. We are proposing experiments that will answer longstanding questions regarding the secretory mechanism of Filamentous hemagglutinin (FhaB), a critical virulence factor for Bordetella pertussis and to identify peptides which inhibit FhaB secretion. FhaB mediates adherence to mammalian respiratory epithelial cells, suppresses the inflammatory response and is required for Bordetella to persist in the murine respiratory tract. FhaB is a TpsA protein secreted by the TpsB protein FhaC. FhaBC are the model for two-partner secretion (TPS) systems. TPS systems are broadly distributed among Gram-negative bacteria and many have proven or postulated roles in virulence. TpsB proteins are part of the Omp85 superfamily which also includes outer membrane protein assembly factors such BamA. Due to the complex topology and large size of FhaB and other TpsA effectors, no structures of active TpsB transporters in complex with their effectors have been solved. The lack of structural information has limited our understanding of how TpsB proteins interact with TpsA passengers and complicates the design of effective anti-microbials targeting these systems. We propose to: 1) Determine the mechanism and structure of the early stages of FhaB secretion by FhaC using a simplified FhaB/FhaC system that we have developed in E. coli and 2) screen a large library (>1012) of cyclic peptides to identify those that bind and inhibit FhaC. This is a collaborative project that takes advantage of the expertise and resources of the Cotter and Doyle laboratories. The results of this work will lay the foundation for a substantial collaboration between the two groups investigating the molecular basis for the TPS mechanism and identification of antimicrobial compounds targeting these systems in Gram negative pathogens.

NIH Research Projects · FY 2026 · 2023-12

Smooth muscle cells (SMCs) play a critical role in atherosclerosis and coronary artery disease (CAD) risk. During disease, a small subset of SMCs proliferate extensively, undergo cell fate change(s) and migrate into the lesion in a process termed phenotypic modulation. Using single cell RNA sequencing (scRNAseq), we have shown that this cell state transition is a continuum in which these SMCs first assume a fibroblast-like phenotype and ultimately transition to a calcific phenotype. We have also shown that top CAD-associated genes modify disease risk by altering the process of SMC phenotypic modulation. Our subsequent preliminary scRNAseq data suggest that different modulated SMC phenotypes might arise as the result of discrete cell fate decisions resulting in different transcriptional paths during disease. This suggests that causal CAD genes function by altering the clonal proliferation and/or cell fate determination of SMCs during disease, but these processes cannot be directly observed with traditional scRNAseq approaches. We have recently shown that the transcription factor TWIST1 is the causal gene at 7p22.1, a genomic locus in human GWAS that is associated with multiple vascular diseases including CAD, stroke, peripheral artery disease and Moyamoya disease. Our preliminary data show that SMC-specific Twist1 knockdown: i) decreases lesion size in the ApoE- /- atherosclerotic mouse model, consistent with the directionality predicted by human GWAS, ii) results in a >60% reduction in the number of SMC-derived cells within the lesion and iii) specifically results in depletion of the calcific modulated SMC phenotype in scRNAseq data. In addition, siTwist1 knockdown in human coronary artery SMCs (HCASMCs) reveals a strong reduction in cellular proliferation/viability and migration. Taken together, these data lead to our central hypothesis that the 7p22.1 locus acts through TWIST1 to drive vascular disease risk by promoting SMC proliferation and altering cell fate decisions during phenotypic modulation, which will be explored through three Specific Aims. Aim 1 will use a novel scRNAseq-compatible cell barcoding mouse model to track individual SMCs as they proliferate and undergo cell fate determination during atherosclerosis, with and without Twist1 knockout. In Aim 2, we will use a SMC-specific myc-tagged Twist1 overexpression mouse model to simulate the human risk allele, and determine whether increased Twist1 expression in SMCs is sufficient to worsen atherosclerosis. We will also leverage the expression of myc-Twist1 to query genome-wide Twist1 DNA binding in SMCs, identifying the direct molecular targets of Twist1 in vivo during atherosclerosis. Aim 3 will determine the cellular pathways and transcription factors that act on the causal SNP at 7p21.1 to regulate TWIST1 expression. Completion of these studies will result in a detailed map of the SMC proliferation and lineage determination landscape during atherosclerosis, which will be a major resource for the field. We will determine how Twist1 alters this landscape, and elucidate the detailed molecular mechanisms of how 7p21.1 and TWIST1 affects the risk for multiple vascular diseases in humans.

NIH Research Projects · FY 2025 · 2023-11

ABSTRACT Obesity is associated with chronic inflammation and an impaired immune response to infection from select viruses, including influenza and SARS-CoV-2, leading to increased morbidity and mortality. Many studies have demonstrated a critical role for T cells in this setting, with primary and memory T cell responses to viral infection impaired in mice and humans with obesity. Given the high prevalence of obesity and viral infections with influenza and coronavirus worldwide, it is critically important to understand T cell dysfunction in obesity and identify novel strategies to improve immune response to infection in this high-risk population. T cell function and metabolism are closely linked, and many studies have demonstrated that changes to T cell metabolism influence T cell fate and function. We have found that activated T cells from obese animals have an altered metabolic profile characterized by increased glucose uptake, increased conversion of glucose to pyruvate, and increased mitochondrial oxidation. This represents a unique cellular metabolic phenotype of glucose oxidation that is not utilized by naive, memory, or activated T cells from lean animals and may mechanistically explain obesity- associated T cell dysfunction. Interestingly, we found that weight loss achieved by continuous low-fat diet was unable to improve obesity-associated inflammation, normalize T cell metabolism, or improve survival to influenza infection in obese mice. Thus, alternative approaches to decrease obesity-associated inflammation and/or restore T cell metabolism may be needed to improve T cell responses to viral infection in individuals with obesity. Alternative dietary approaches using variations of time-restricted feeding such as intermittent calorie restriction, intermittent fasting, or alternate day fasting have been shown to be beneficial to both weight loss and multiple indices of health, including metabolic disease, inflammation, and immune response to bacterial infection. Recent publications suggest that the immunomodulation seen following time-restricted feeding is at least partially mediated by changes in the gut microbiome. Indeed, it is now well established that the gut microbiome of mice and humans with obesity is different than the gut microbiome of lean animals, and it is the prevailing view that these changes in gut microbiome in obesity drive adipose and system inflammation and thereby influence immune cell responses. Therefore, the overall objective of this R21 proposal is to determine the effects of time- restricted feeding on gut microbiome diversity, T cell function and metabolism, and survival to influenza infection. We hypothesize that weight loss achieved through time-restricted feeding will alter the gut microbiome and reduce obesity-induced inflammation, thereby restoring T cell metabolism and function, resulting in decreased morbidity and increased survival to influenza infection. Successful completion of these studies will identify metabolic and microbiome-related mechanisms underlying T cell dysfunction in obesity and test the impact of alternative dietary approaches on T cell response to viral infection.

NIH Research Projects · FY 2026 · 2023-11

Population aging suggests the need for an increased workforce of community advocates, academic scholars, health care providers, and other professionals with training in gerontology/geriatric medicine or a related discipline. Established in the early 19th century, Historically Black Colleges and Universities (HBCUs) were created to educate Black students when they were not allowed to attend certain public and/or private predominately white institutions. With increasing enrollment, HBCUs are now being recognized for their progressive academic presence in the Black community educating distinguished scientists, inventors, politicians, and world leaders. These institutions have similarly become increasingly integrated into the aging network by educating (undergraduate/graduate) some of the most influential gerontologists and geriatric practitioners. Despite the role of HBCUs in educating and preparing established and emerging scientists, there remains limited professional development opportunities, a lack of scholarly resources, and inadequate mentoring and networking supports for HBCU scientists by established gerontologists. To address these issues, scholars will collaborate to organize and host conferences for emerging and early career scientists from HBCUs. The aims of the HBCU aging conference series are to: host five multidisciplinary annual conferences; address knowledge gaps and identify future priorities in gerontology related to aging; prepare scientists for leadership positions/roles in gerontology; facilitate professional development, mentoring, and networking opportunities within and between HBCUs, and other community-based and academic institutions; and sponsor a select number of emerging and early career scientists from HBCUs to attend and participate in the annual conference through the HBCU Aging Ambassadors Program (HAAP). Each conference will be an interactive two-day pre-conference workshop during the GSA’s annual scientific meeting. All attendees will be engaged with panel discussions, professional development breakout sessions, and facilitated mentoring and networking opportunities. This conference series will serve as an essential hub for knowledge transfer and exchange, professional skills and practice improvement, sustainable best practices in gerontology, proficiency in established competencies, and transformation in the quality of aging scholarship. The novelty of these conferences is that, to our knowledge, it is the first to focus exclusively on scientists engaged in gerontology or a related discipline from HBCUs.

NIH Research Projects · FY 2025 · 2023-11

ABSTRACT Depressive and anxiety disorders are among the leading contributors to the global disease burden and are more common in women compared to men. Among women in low-resource contexts, such as low- and middle- income countries (LMICs), exposure to adversities (e.g., poverty, intimate partner violence, negative life events such as illness or loss) is a powerful risk factor for developing internalizing symptoms (i.e., depressive and anxiety symptoms). Dysregulated activity in the hypothalamic pituitary adrenal (HPA) axis may be a mechanism underlying the onset of internalizing disorders and a novel adjunct target for mental health interventions in addition to psychological symptoms. However, there is little evidence of the long-term effects of mental health interventions on either psychological symptoms or HPA axis dysregulation. In addition, more research is needed to characterize the dynamic role of adverse experiences in shaping women’s internalizing symptoms and HPA axis activity over time, especially in South Asia, where women may have unique familial social contexts compared to western counterparts. This work is essential for establishing and improving the long-term effectiveness of mental health interventions globally. In this R21, we propose leveraging multi-year experimental and observational data from the Bachpan study, an ongoing longitudinal birth cohort with an embedded cluster randomized control trial of a maternal depression intervention delivered in rural Pakistan. Specifically, we will test the effects the intervention on HPA axis regulation and internalizing symptoms 6 years after intervention onset, as well as whether HPA axis activity mediates the effect of the intervention on internalizing symptoms. We will also examine how adversity exposure, including chronic adversity experienced over a six-year period, retrospectively reported childhood adversity, and recent adversity, relate to HPA axis activity and internalizing symptoms. As part of this analysis, we will also examine how within-person changes in adversity relate to within-person changes in HPA axis activity and internalizing symptoms over time. The rich, multi-year dataset includes information on multiple domains of women’s mental health and adverse experiences, as well as hair-derived HPA axis hormones (cortisol and DHEA). The major public health contribution of this project is that it will generate knowledge on the longitudinal predictors of stress system dysregulation and internalizing symptoms among women in a low resource context, as well as the potential ameliorating role of psychosocial intervention.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Inflammatory bowel diseases (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), are chronic, relapsing and debilitating disorders without a cure. IBD affects more than 3 million individuals in the United States, including people of all ages, sex, race and socioeconomic status. There is a critical need to better define the role of social determinants of health on the course of IBD and to identify ways to mitigate health disparities. The overarching goals of this research program are to improve quality of life and reduce health disparities through: 1) rigorous epidemiological research to evaluate impact of social risk factors on the natural history of IBD, 2) testing the effectiveness of innovative IBD management strategies to improve health equity and outcomes, and 3) developing and evaluating stakeholder-informed patient and provider education and awareness programs. The project will consist of a 52-week, pragmatic, multi-center, open-label, randomized clinical trial with a prospective cohort study embedded within the trial. We will include 1200 children (13-17y) and adults (>17y) with IBD at 6 pediatric and 5 adult sites. Aim 1 is a prospective cohort study (n=800) that will be comprised of participants assigned to the usual care arm of the pragmatic trial. The aim is to evaluate the association between social risk and natural history and outcomes of IBD in children and adults, Aim 2 will involve the development of a multi-stakeholder informed, tailored digital health intervention to improve IBD management and patient activation followed by a pragmatic trial to compare the effectiveness of the intervention versus usual care. Aim 3 will address health disparities and increase IBD awareness through provider and patient education and the dissemination of study findings and other evidence-based practices to researchers, clinicians, and patients. This work will bring together a coalition of professional organizations, medical education events, disease- associated non-profits and grassroot patient advocacy groups representing diverse patients with IBD who are to create, conduct and evaluate live, in-person and digital education and awareness programs.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY/ABSTRACT Legalization of cannabidiol (CBD) in 2018 opened the door to intentional chemical conversion of CBD to Δ8- tetrahydrocannabinol (Δ8-THC), an unregulated psychoactive cannabinoid. Vaping products containing Δ8-THC have since rapidly gained popularity with consumers; however, little is known about their respiratory health effects. There is an urgent need to evaluate the effects of vaping Δ8-THC as the recent outbreak of e-cigarette and vaping product associated lung injury (EVALI) included Δ8-THC vaping product users. Additionally, vaping- induced thermal oxidation processes can convert Δ8-THC into reactive electrophiles, including Δ8-THC quinone and CBD quinone (Δ8-THCq and CBDq), which may cause lung injury. Our long-term goal is to enhance the health of all individuals by providing the public with accurate safety information about vaping Δ8-THC and guiding regulatory decision making on Δ8-THC and other cannabinoids. The objective of this proposal is to understand how vaping Δ8-THC alters normal lung cell function and the mechanism mediating these effects. The central hypothesis is that aerosolized Δ8-THC vaping liquids containing pro-oxidant impurities generate reactive Δ8- THCq and CBDq, which adduct protein thiol residues and impair normal biological pathways in human airway epithelial cells. Our specific aims will determine (aim 1) how aerosolized Δ8-THC vaping products alter normal airway epithelial cell function, (aim 2.1) how impurities including metals and flavoring chemicals affect the oxidation of Δ8-THC to quinones Δ8-THCq and CBDq, and (aim 2.2) airway epithelial cell protein adducts formed from vaping Δ8-THC. The results of this proposal will be significant as they will identify for the first time the effects of aerosolized Δ8-THC on normal airway function and the vaping-induced Δ8-THC “adductome”. This project is innovative in its use of (1) a novel in vitro Vaping Product Exposure System (VaPES) to assess the effects of Δ8- THC aerosols on primary human bronchial epithelial cells grown at air-liquid interface and (2) alkynyl-tagged cannabinoids and click chemistry methodologies to uncover the “adductome” of proteins covalently modified by Δ8-THC oxidation products. This proposal will provide training opportunities in technical skills including chemical analysis of vaped condensates, click chemistry, and proteomic analysis, which will be critical for my development as an independent scientist. Finally, I will enhance my training in science communication and the impact of this work on the health of our community by translating the findings of this proposal into a lesson plan that I will implement at local middle and high schools.

NIH Research Projects · FY 2024 · 2023-09

Abstract Cannabis (CB) use is prevalent amongst people with HIV (PWH) but its impact on HIV infection and the latent viral reservoir has not been fully examined. In particular, the impact of CB on the HIV reservoir in solid tissues such as the brain is completely unknown. As such, animal models in which dosing can be carefully controlled and tissue samples can be analyzed will be important to fully understand how CB impacts HIV reservoir characteristics in tissues. Our central hypothesis is that CB induces an immunosuppressive cellular state that impacts viral transcription and maintenance of the HIV reservoir in tissues. The overall goal of this proposal is to develop a humanized mouse model of CB exposure during HIV infection and to use this model to learn how CB use affects characteristics of the tissue resident HIV reservoir. In the R61 phase, HIV infected humanized mice expressing hIL-34 to promote microglial CNS reconstitution will be exposed to a range of THC doses during infection, both before and after viral suppression with antiretroviral therapy (ART). The impact of THC exposure on viremia, viral suppression, immune markers and cytokines in peripheral blood will be examined. In the R33 phase, we will use this model to address three major questions. First, viral DNA (vDNA) and viral RNA (vRNA) levels in the brains and spleens of infected mice that have been dosed with THC will be quantified to determine the impact of THC on the size and transcriptional activity of the viral reservoir in these tissues. Second, we will examine whether THC exposure affects HIV latency reversal in vivo. We hypothesize that cross-talk between inflammatory signaling pathways activated by potent latency reversing agents (LRAs) such as the non-canonical NF-kB agonist AZD5582 and anti-inflammatory pathways activated by CB use could significantly alter the outcome of LRA dosing in vivo. To test this hypothesis, THC or control exposed HIV infected mice will be dosed with AZD5582, and the impact of THC on viremia and viral reactivation in tissues will be examined. Finally, we will use this model to perform a deep single cell characterization of infected cells in the brains and spleens of ART-suppressed humanized mice in the presence and absence of THC exposure. Spleen and brain tissues will be profiled using a combined single cell ATACseq/RNAseq method to identify and characterize HIV vDNA+ or vRNA+ cells and to compare the abundance and phenotype of these cells between CB dosed mice and controls. If successful, this proposal will reveal new insights into how CB affects HIV infection and the characteristics of the tissue-resident HIV reservoir. This in turn will guide novel HIV cure strategies that are optimized for CB-using PWH.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT: Violent deaths are responsible for the most person-years of life lost among people 18 to 50 years old in the United States. Further, violence inflicts a disproportionate burden on racial and ethnic minorities in the US, thereby exacerbating existing health inequities, including those amplified by COVID-19. Although the physical distancing policies put in place to address the COVID-19 pandemic reduced social interactions, violence-related injury and homicides increased in 2020 in the US. Structural and policy interventions that target the social-ecological framework can attain sustainable reductions in multiple forms of violence. Given that poverty is one of the drivers of violence, increasing minimum wage may be a potent policy intervention for violence prevention. However, minimum wage increase laws have not been evaluated as a community violence prevention method. Further, several states increased minimum wage during 2020 and 2021, providing a unique opportunity to examine whether such increases can prevent community violence during the pandemic. Our study will address three important questions: 1) what role do minimum wage increases play in community violence prevention? 2) Does minimum wage increase or lack thereof differentially affect community violence among different demographic groups (race/ethnicity, sex, age, and rurality)? and 3) how has the COVID-19 pandemic impacted community violence and if state minimum wage (SMW) increases and economic impact payments (EIP) played a role in mitigating pandemic effects? To address these research gaps, our team will employ rigorous quasi-experimental designs (synthetic controls and controlled interrupted time series) with quantitative bias analyses, on six national datasets from 2000-2021 to capture community violence victimizations, assault injuries, and homicide deaths in all 50 states. We will then examine the interaction of minimum wage with income inequality measured using living wage. We will estimate the impacts of the COVID-19 pandemic itself, and the impact of state level increases in minimum wage during the pandemic on community violence outcomes. Through all these analyses we will examine disparities by race, sex, age, and rurality. This study is responsive to the RFA-CE-23-004's funding objective three to evaluate approaches that improve social or structural contributions that contribute to community violence and racial/ethnic inequities. This study will be the first to comprehensively assess the impact of minimum wage increases on violent victimizations, injuries, and homicide deaths and the racial, sex- and age-based, and geographical disparities therein. Further, this study will also be the first to examine how SMW increases and EIPs may have moderated the impact of COVID-19 pandemic on community violence in the US. We will employ state-of-the-art epidemiologic methods to build a strong evidence base to control and reduce the burden of community violence in the US. Understanding the impact of SMW increases, which greatly affect racialized minorities and women, will also provide guidance to reduce systemic inequities.

NIH Research Projects · FY 2024 · 2023-09

Project Summary/Abstract: Violence is a common, costly, and preventable cause of morbidity and mortality for children in the United States. Violence against children occurs more frequently for children who grow up in poverty. The primary method of addressing poverty in the U.S. is the provision of social services. Yet, these safety net services underserve the communities that are the most in need of them. This mismatch between safety net need and accessibility is a structural condition that may be a social determinant of health related to violence against children. Rigorous research, such as causal effects study designs, is needed to identify modifiable community- level factors for targeted prevention. The proposed Career Development Award (K01) research goal is to evaluate the relationship between the community safety net accessibility, the primary method of alleviating poverty, and violence against children. The proposed study leverages population-based, longitudinal data from multiple sources, including novel linked administrative data and detailed census-tract-level data on social service availability. The proposed research activities will address the following specific aims: 1) determine, at the community level, if neighborhood safety net accessibility is related to violence against children indicators (child protection reports, child protection placements, violence-related medical encounters, and violence- related deaths); and 2) quantify, at the child-level, the causal impact of community safety net accessibility on violence-related mortality through the duration of childhood. To pursue these aims, the PI will receive training in rigorous causal effects study design and cross-cutting violence prevention research under the mentorship of national leaders in these areas. The proposed research study addresses the important research priority for the CDC’s National Center for Injury Prevention and Control of the relationship between community conditions and violence against children using an innovative approach with a strong scientific premise. Completion of the training and research activities will allow the PI to become an independent researcher who produces high- impact research that can be used in the development of prevention programming and to better explain the structural conditions that contribute to violence against children.

NIH Research Projects · FY 2024 · 2023-09

NIH Research Projects · FY 2025 · 2023-09

The US has made decades of progress in reducing cigarette smoking, but there are notable differences in menthol cigarette smoking and use of non-cigarette tobacco products including little cigars and e-cigarettes. We propose the U54 Program Project, “UNC Center of Tobacco Regulatory Science” to address this gap. The Integrative Theme of our UNC TCORS is building the science for effective public health decision-making about flavored tobacco products. Our unified set of four Research Projects seek to understand the impact of tobacco-control and messages on tobacco use. Our goal is to further FDA and NIH efforts to protect public health through tobacco control decision making. UNC TCORS’ research will provide essential evidence to guide the FDA as it implements restrictions on menthol cigarettes and flavored cigars, develops messages, and addresses the youth vaping epidemic. We propose four Specific Aims: 1) Develop and evaluate the impact of messages to discourage tobacco product use (Projects 1, 2, 4); 2) Understand the public health impact of flavored tobacco product sales restrictions and the conditions under which they are most likely to be most effective (Projects 2 and 3); 3) Build trainees’ capacity to contribute to the field of tobacco regulatory science through training, mentorship, and pilot funding for a multidisciplinary group of pre- and post-doctoral fellows and early-stage investigators (Projects 1-4, Career Enhancement Core); 4) Inform FDA tobacco-related decision making of menthol cigarette and flavored cigar sales restrictions and messages through active dissemination of TCORS scientific findings (Projects 1-4, Administrative Core). Project 1 will develop novel digital messages to discourage young adult little cigar and cigarillo use. Project 2 will evaluate whether ending the sale of menthol cigarettes could be amplified by a quit smoking campaign to help menthol smokers quit. Project 3 will build a microsimulation model to estimate the public health impact of a federal flavored cigar sales restriction on tobacco use, mortality, and health. Project 4 will test the ability of vaping prevention video ads with promising features to reduce susceptibility to vaping among youth and young adults. The Research Projects will receive support from two Cores: Administrative and Career Enhancement (which includes Pilot Research Projects). Our multidisciplinary group of seasoned regulatory science researchers will provide actionable information to inform FDA tobacco control decision making and messaging strategies.

NIH Research Projects · FY 2025 · 2023-09

Abstract Cannabis (CB) use is prevalent amongst people with HIV (PWH) but its impact on HIV infection and the latent viral reservoir has not been fully examined. In particular, the impact of CB on the HIV reservoir in solid tissues such as the brain is completely unknown. As such, animal models in which dosing can be carefully controlled and tissue samples can be analyzed will be important to fully understand how CB impacts HIV reservoir characteristics in tissues. Our central hypothesis is that CB induces an immunosuppressive cellular state that impacts viral transcription and maintenance of the HIV reservoir in tissues. The overall goal of this proposal is to develop a humanized mouse model of CB exposure during HIV infection and to use this model to learn how CB use affects characteristics of the tissue resident HIV reservoir. In the R61 phase, HIV infected humanized mice expressing hIL-34 to promote microglial CNS reconstitution will be exposed to a range of THC doses during infection, both before and after viral suppression with antiretroviral therapy (ART). The impact of THC exposure on viremia, viral suppression, immune markers and cytokines in peripheral blood will be examined. In the R33 phase, we will use this model to address three major questions. First, viral DNA (vDNA) and viral RNA (vRNA) levels in the brains and spleens of infected mice that have been dosed with THC will be quantified to determine the impact of THC on the size and transcriptional activity of the viral reservoir in these tissues. Second, we will examine whether THC exposure affects HIV latency reversal in vivo. We hypothesize that cross-talk between inflammatory signaling pathways activated by potent latency reversing agents (LRAs) such as the non-canonical NF-kB agonist AZD5582 and anti-inflammatory pathways activated by CB use could significantly alter the outcome of LRA dosing in vivo. To test this hypothesis, THC or control exposed HIV infected mice will be dosed with AZD5582, and the impact of THC on viremia and viral reactivation in tissues will be examined. Finally, we will use this model to perform a deep single cell characterization of infected cells in the brains and spleens of ART-suppressed humanized mice in the presence and absence of THC exposure. Spleen and brain tissues will be profiled using a combined single cell ATACseq/RNAseq method to identify and characterize HIV vDNA+ or vRNA+ cells and to compare the abundance and phenotype of these cells between CB dosed mice and controls. If successful, this proposal will reveal new insights into how CB affects HIV infection and the characteristics of the tissue-resident HIV reservoir. This in turn will guide novel HIV cure strategies that are optimized for CB-using PWH.

NIH Research Projects · FY 2026 · 2023-09

PROJECT SUMMARY A key priority for the NIH is to limit disability caused by osteoarthritis (OA) and other chronic diseases that emerge with age. Senescent cells within joint tissues contribute to OA, but there is a knowledge gap regarding the triggers by which decades of aging initiate cellular senescence. One key mediator of senescence in other contexts is persistent DNA damage and the subsequent activation of a set of signaling pathways known as the DNA damage response (DDR). The DDR can drive the production of inflammatory and matrix-degrading molecules collectively known as the senescence-associated secretory phenotype (SASP), which has strong overlap with catabolic molecules known to contribute to OA. As demonstrated through the use of a single-cell gel electrophoresis “comet” assay, chondrocytes accumulate significant levels of DNA damage throughout aging and during OA. This damage is mostly in the form of single-strand breaks (SSBs) but a subset of cells also harbor double-strand breaks (DSBs). These distinct forms of damage can be initiated in cells from young cadaveric donors and mice to mimic the levels found in older donors/mice, with methyl methanesulfonate (MMS) for SSBs, ellipticine for DSBs, and irradiation to generate both SSBs and DSBs. Conversely, the burden of DNA damage in older cadaveric donors and older mice can be reduced by boosting DNA repair with activation of Sirtuin 6 (SIRT6) using the small molecule MDL-800. The long-term goal of this work is to catalyze more effective treatments for OA by determining the mechanisms by which joint cells become senescent. The central hypothesis is that the accumulation of DNA damage in joint tissues plays a causal role in driving senescence, the SASP, and subsequent OA. The first aim is to establish the contribution of SSBs and DSBs to senescence by applying distinct forms of DNA damage (irradiation, MMS, ellipticine) to cadaveric human chondrocytes and synovial cells. The second aim is to determine the extent to which DNA damage drives senescence and OA in mice. We also use intra-articular injection of agents to increase damage (MMS or ellipticine) or decrease damage (MDL-800) in the joints of p16tdTom reporter mice to assess senescence and functional/histologic OA. The third aim is to define the protein signatures that contribute to progression towards the SASP using a multiplex antibody staining method known as iterative indirect immunofluorescence imaging (4i) to track the signaling pathways that are activated in response to DNA damage. The expected outcomes of this work include a better understanding of the types of DNA damage that lead to senescence in joint tissues and the signaling pathways that link the DDR to SASP. This work is innovative in that tailored interventions are employed to alter the levels of DNA damage, with sophisticated readouts of senescence, including 4i for assessing protein signatures and senescence reporter mice. These contributions are expected to have a positive impact on society by stimulating more effective strategies to target senescent cells for the prevention and treatment of OA.

NIH Research Projects · FY 2023 · 2023-09

Autosomal dominant polycystic kidney disease (ADPKD) is the most common potentially lethal genetic disease. ADPKD is caused mainly by mutations in the PKD1 gene, which encodes the polycystin-1 (PC1) protein. Therapeutic treatment of ADPKD that targets the proximal signaling functions of PC1 has yet to be discovered. PC1 is an important unusual G-protein-coupled receptor (GPCR) with 11 transmembrane (TM) domains. PC1 shares multiple characteristics with Adhesion GPCRs. These include a GPCR proteolysis site that autocatalytically divides these proteins into extracellular, N-terminal and membrane-embedded, C-terminal (CTF) fragments. A tethered peptide agonist (TA) within the N-terminal stalk of the CTF has been suggested to activate signaling of PC1. Using the cryo-EM structure of PC1, we have recently revealed a novel allosteric TA/stalk-mediated signaling mechanism of PC1 by combining complementary all-atom Gaussian accelerated molecular dynamics (GaMD) simulations and biochemical and cellular assay experiments. Moreover, we have uncovered unique features of activation and allosteric modulation in the A and B classes of GPCRs from sequence coevolutionary “Potts” models and structural contact analysis. We have shown how “Potts” models fit to homologous sequences can be used to generate and detect cryptic functionality of multiresidue sequence motifs involved in allosteric binding and signaling. In addition, we have developed the GaMD, Deep Learning and free energy prOfiling Workflow (GLOW) to predict molecular determinants and map free energy landscapes of functional biomolecules. Building upon these advances, we will design and test novel peptide modulators to probe mechanisms of PC1 signaling regulation by combining state-of-the-art computational techniques (including sequence coevolutionary Potts models, GaMD, GLOW and peptide docking) and complementary cellular signaling experiments. Our specific aims include: (1) Characterize the binding mechanisms of known TA/stalk-derived peptide modulators of PC1 through sequence coevolution analysis, peptide docking, and AI modeling; and (2) Predict and validate new peptide modulators of PC1 through Potts modeling, peptide virtual screening, and cellular signaling assays. Therefore, we will implement a unique computational sequence- and structure-based learning approach coupled with relevant in vitro experimental analyses to develop novel peptide modulators of PC1. Our long-term goals are (1) to develop robust computational and experimental methodologies to characterize protein-peptide interactions, (2) to understand mechanisms of signaling in the wildtype and ADPKD disease mutants of PC1, and (3) to lay the foundation for the future design of effective therapeutics for treatment of ADPKD.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT Extreme weather events pose a growing threat to the health and development of vulnerable populations in the United States. Psychological distress from extreme storms often persists for years, air pollution from wildfires increases the risk of adverse birth outcomes, and floods spread contaminants in land and water. While the threats from chronic extreme weather to wellbeing are clear, research has predominantly focused on the effects of extreme events, rather than how the “new normal” under chronic extreme weather and its effects broadly can cause frequent disruptions and long-term stress for households and communities. For many coastal communities, sea level rise is creating a new normal with routine flooding from monthly high tides and rain. While those floods are not as catastrophic as hurricanes, their near-certain occurrence causes different problems – and potentially leads to different responses – than low probability hurricane strikes. Understanding the impacts of these chronic stresses, alone and in combination with acute disasters, is necessary for enabling effective adaptation. This K01 Award proposal will advance our understanding of the impacts of chronic and acute coastal flooding on migration and stress. By building on my expertise in exposures and impacts, it will accelerate my progress towards my career goal of becoming an independent researcher at the intersection of extreme weather and population science. I will train in demographic theory and methods, conduct guided readings with my mentors on measurement of psychosocial and physical health, and develop survey design and analysis skills. Then, I will apply those skills to study the interactions between flooding, migration, and health in coastal communities in the US. My research aims are: 1) to characterize the relationship between flood experience and in- and out-migration, 2) to analyze how health, economic resources, and flood experience shape migration intentions, and 3) to assess the health and economic wellbeing of movers and stayers. To achieve these aims, I will draw on two primary datasets: an administrative dataset consisting of individual address histories for most adults in eight Atlantic coast states, and a detailed household survey currently underway in four coastal North Carolina counties. My mentor team, with expertise in environmental migration, survey methods, and health measurement, are ideally suited to help me achieve my research and training goals. The outcomes of the research and data collected in this study will form the basis for future R-level proposals to expand the geographic scope of the household survey on the impacts of coastal flooding and to examine the health and development impacts of sea level rise on children and young adults.

NIH Research Projects · FY 2025 · 2023-09

Abstract The primary route of internalization from the plasma membrane is clathrin-mediated endocytosis (CME), which is required for vital cellular processes such as signaling, nutrient uptake, and development. While the molecular mechanisms underlying the late stages of vesicle formation are well studied, the mechanisms underpinning the earliest stages of endocytosis, including initiation and cargo selection, are poorly understood. Initiation and cargo binding are largely controlled by the AP2 clathrin adaptor complex, a core component of the vesicle coat that serves as a bridge between the rigid clathrin lattice and membrane-embedded cargo. However, it is unclear how AP2 discriminates between hundreds of potential cargo in a complex membrane environment, while also responding to spatial and temporal regulatory cues. While >50 proteins are proposed to regulate or be required for CME, and many physically interact with AP2, we have little mechanistic and structural data for how they are regulated during the earliest stages of endocytosis. We propose that multiple unique AP2 conformations, driven by association with regulatory factors, control higher-order AP2 functions such as cargo selection. Understanding the nature of the regulatory mechanisms controlling endocytosis is critical, as CME largely controls the localization of many medically-relevant proteins such as RTKs and GPCRs. This proposal therefore seeks to reconstitute and define the molecular mechanisms of AP2-mediated endocytic initiation and cargo sorting. Current models of endocytosis largely rely on biochemical experiments performed with soluble components and live cell imaging. Importantly, our methodology is focused on modifying all experimental approaches — cryo-EM, biochemical reconstitution, and single molecule fluorescence microscopy — to include a membrane, thereby addressing a critical need to develop mechanistic models in a near-native membrane environment. This approach is poised to provide an understanding of the role of membrane-induced allostery in driving regulatory decision-making during endocytic initiation. This proposal will focus on two broad areas of endocytic regulation — cargo selection mediated by the conserved Muniscin family proteins, and a quality control checkpoint controlled by a single phosphorylation mark on the μ2 subunit of AP2. As diseases associated with endocytic defects are likely caused by missorting of important trans-membrane cargo, our insights into the mechanisms of endocytic initiation and cargo selection will enable hypothesis-driven research into disease model systems and drug development.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT While the principal function of tumor suppressor p53 is a nuclear transcription regulator that transcribes a diverse set of genes involved in apoptosis, cell cycle regulation, and numerous other processes, compelling evidences have also unraveled extra-nuclear, transcription-independent activities of p53 in the cytoplasm, where it induces apoptosis and inhibits autophagy. However, significant gaps exist in the knowledge of p53 in the cytoplasm. The lack of knowledge is due at least in part to the fact that approaches investigating cytoplasmic p53 have been primarily in vitro studies in artificial settings. Dissection of the mechanistic aspects of cytoplasmic p53 in an in vivo system is necessary to clarify the biological events by which cytoplasmic p53 exerts its extra-nuclear, transcription-independent functions. Recent genome sequencing analyses of bats discovered many unexpected variances between bats and other mammals. Among them are the bat-specific alterations in the nuclear localization signal (NLS) sequences in p53. The alterations are predicted to hamper bat p53 nuclear import, resulting in increased cytoplasmic accumulation. To explore the significance of cytoplasmic p53, we generated mice expressing a mutant p53 that has an altered NLS sequence to mimic bat p53 NLS. We identified that p53 interacts with lactate dehydrogenase B (LDHB), which plays a critical role in lipid metabolism. We also found that cytoplasmic p53 promotes autophagy through disrupting Beclin 1-Bcl-2/Bcl-xL interaction and activating Beclin 1. IP-mass spectrometry experiments using cytoplasm-sequestered p53 and identified CUL9, a cytoplasmic cullin RING E3 ubiquitin ligase, as a major p53 binding partner. Biochemical experiments revealed that CUL9 catalyzes K63-linked, non-proteolytic polyubiquitination of p53. In this proposal, we test the hypothesis that cytoplasmic p53 exerts tumor suppression and anti-aging functions through inhibiting glycolysis, increasing lipogenesis, and promoting autophagy. Such functions of p53 are regulated by CUL9 catalyzed, K63-linked p53 polyubiquitination.

NIH Research Projects · FY 2025 · 2023-09

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Chronic kidney disease affects one in seven U.S. adults and is a leading cause of death and disability. Chronic kidney disease has a substantial impact on U.S. populations. Research on the epidemiology and molecular risk factors contributing to chronic kidney disease in US populations is much needed to guide public health prevention and interventions, and to develop therapeutic tools for clinicians to effectively treat the disease. This K26 application will support protected time for Nora Franceschini, MD MPH, to mentor junior investigators on molecular epidemiology research of chronic kidney disease. Dr. Franceschini is a nephrologist and an epidemiologist with a 20-year experience conducting multidisciplinary studies in chronic kidney disease and mentoring graduate students and early-stage investigators, including physicians and PhD candidates. Dr. Franceschini has developed a strong National Institutes of Health supported research program on chronic kidney disease at the University of North Carolina focused on global populations. This research program offers multiple opportunities for trainees to learn rigorous research skills in state-of-art approaches for research in the epidemiology, molecular epidemiology and biomarkers of chronic kidney disease. Proposed activities and enhanced mentoring skills support the goals to increase high-quality mentoring for the next generation of scientists in chronic kidney disease research within her research program.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY This goal of this K23 Mentored Patient-Oriented Research Career Development Award is for the candidate to gain expertise in childhood threat exposure, social motivation (SM), and risk for suicidal thoughts and behaviors (STBs). Childhood threat exposure increases risk for STBs by affecting brain functioning associated interpersonal stress. Adolescence is filled with interpersonal stress; yet, not all adolescents with childhood threat exposure experience STBs, indicating that additional factors exacerbate risk. Identifying additional factors that interact with threat to increase STBs is imperative for identifying novel suicide prevention target mechanisms. This project will identify whether SM represents an additional factor that moderates the link between childhood threat exposure and STBs. Training objectives for this award will include skill acquisition in: (1) Developmental psychopathology models of childhood threat exposure and SM, (2) Neurobiological and behavioral mechanisms of risk for STBs, and (3) Advanced intensive time series modeling of brain connectivity. Training will facilitate the ability to conduct a study characterizing self-report, brain, and behavioral measures of SM in adolescents (Aim 1) to examine whether maladaptive social motivation strengthens the link between threat exposure and STBs (Aim 2). A final exploratory Aim 3 will allow for a preliminary investigation into how puberty, biological sex, and age further moderate the interaction between threat exposure and SM on STBs. This study will leverage two R01 projects to test Aims in two Cohorts; Cohort 1 (n = 220) includes early adolescent females aged 9 – 16, while Cohort 2 (n = 275) consists of adolescent male and females aged 15- 18. Experts from the University of North Carolina at Chapel Hill, Washington University St. Louis, and the University of Illinois Urbana-Champaign will mentor and consult with the candidate on the 3 training goals of this project: Margaret Sheridan Ph.D. & Adam Miller Ph.D. (Co-primary mentors), along with Karen Rudolph Ph.D., Deanna Barch Ph.D., Katie Gates, Ph.D. (Scientific Advisors), and Eva Telzer, Ph.D. and Joan Luby, M.D. (Consultants). This award provides the necessary training to become a unique, transdisciplinary researcher at the intersection of clinical, social, and developmental neuroscience. In consultation with the mentors, the candidate will use the training and the results of the research to inform the next steps in the candidate’s program of research on childhood adversity, social motivation, and risk for STBs during adolescence. Training and results from this award will also inform future R01 submission.

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract Alcohol (ethanol) exposure during pregnancy is the leading environmental cause of birth defects and central nervous system dysfunction. While the effects of ethanol on the brain and face have been explored quite extensively, there is considerable variation in the consequences of developmental ethanol exposure. Some of this variation is due to differences in timing and amount of exposure or nutritional factors. However, even when controlling for these factors, it is still clear that not everyone exposed to ethanol during development is affected equally and it has become increasingly clear that genetic factors play a very significant role in fetal alcohol spectrum disorders (FASD). Historically, elucidating these genetic factors that mediate risk or resilience has been relatively slow and characterized by human association studies or quantitative trait loci mapping in animal models. More recently, we have applied next generation sequencing technologies and forward genetic screens to more rapidly identify genes and pathways that alter susceptibility to prenatal ethanol exposure. These studies have taken advantage of varied mouse strains with differential alcohol susceptibility, numerous transgenic mouse lines, and high throughput zebrafish genetic analyses and CRISPR/Cas9 gene editing techniques. In this current proposal, we will further these approaches with the combination of the powerful genomic analyses capabilities of the Collaborative Cross mouse genetics tools. In Aim 1, we will explore mechanisms underlying ethanol sensitivity using complementary mouse and fish transgenic lines, while identifying further candidate genes via comparisons of the highly ethanol susceptible mouse strain, C57BL/6J vs. the highly ethanol resistant strain 129S1/Svlmj. This comparison will be aided by embryonic transcriptomic (RNA-Seq) analyses, selective crossbreeding to induce susceptibility in a resistant line (129) with extensive genome sequencing analyses. Aim 2 will significantly expand our genomic analyses by examining CC founder strains for their susceptibility to ethanol followed by transcriptomic profiling of susceptible and resistant strains. Conserved candidate genes and pathways will be further tested and characterized in our high throughput zebrafish phenotyping analyses. Aim 3 will take a complementary bioinformatic approach by identifying chemical modifiers of gene-ethanol interactions in a high throughput zebrafish screen with further confirmation in our mouse model of FASD. This proposal brings together experts on mouse and fish genetics, embryology and alcohol teratology. Together, these experiments will greatly enhance our understanding of the genetic etiology of ethanol-induced brain and craniofacial malformations during early embryonic development, as well as aiding in the identification of the pathogenic mechanisms involved in FASD.

NIH Research Projects · FY 2025 · 2023-09

Project Summary Exosomes, also referred to as small extracellular vesicles, play important roles in cellular communication under physiological and pathophysiological conditions. Exosomes contain a wide range of both short and long non- coding RNAs that regulate many aspects of gene expression including epigenetic processes that modulate cellular fate, phenotype, polarization, and morphogenesis. Despite the important functional roles played by exosomal RNAs, there are currently no methods that allow live exosomal RNA tracking. This is because RNA is by nature non-fluorescent and difficult to label while maintaining its intended biological function. Access to exosomal RNA is further complicated by the fact that each RNA species is present at extremely low copy numbers in exosomes. This emphasizes both the need for a novel marker capable of tracking the intercellular movement of exosomal RNA, and the need to enhance loading of RNAs into exosomes. Given the central importance of exosomal RNAs in dictating cellular behavior, there is a need and demand for exosomal RNA imaging methods to determine how (a) cells use exosomes and their cargoes to communicate with each other and (b) how exosomes modulate their microenvironment and travel to distant organs and tissues. Existing methods focus on tracking exosomes by labeling the lipid membrane via a lipid-based fluorophore or exosomal protein labeling. None of these methods allow the tracking of exosomal RNA via genetic encoding or barcoding without exogenously modifying the exosomes after extensive collection and alteration steps. The overall goal of this proposal is to develop genetically encodable RNA EXO-Code probes that allow multimodal tracking and imaging of exosomal RNAs. The EXO-Code probe will allow multimodal tracking of exosomal RNA via (1) genetic encoding, (2) non-destructive labeling with fluorescent dyes, and (3) unique identification and quantification based on barcoding. This combination is powerful as it allows tracking of exosomal RNA via multiple modes for high content biodistribution mapping. Because EXO-Code barcodes are composed of unique nucleotide sequences, they can be accurately decoded using sequencing with sensitivity in the attomolar range. The combination with a fluorogenic RNA aptamer allows for complementary tracking of exosomal RNA via simple incubation with dyes. The fluorescent exosome toolkit will be developed for investigators to detect disruptions in membrane stability, exosomal fusion events, and endocytic processes as a result of exosome biogenesis, distribution, and uptake. This will enable researchers to track exosomal RNAs through organisms, cells, and their ultimate destinations within subcellular compartments.

NIH Research Projects · FY 2025 · 2023-09

This is a training program from the University of North Carolina at Chapel Hill (UNC-CH) to train postdoctoral fellows in basic, translational or clinical research focusing on tumor immunotherapy, tumor immunology or stem cell transplantation immunology. The UNC Immunotherapy Training Grant (IM-TAG) is led by an exceptional mentor, Jonathan Serody MD, who is assisted by two superb Associate Directors in Barbara Savoldo MD PhD and Benjamin Vincent MD. This training proposal is supported by a nationally renowned mentorship group and exceptional facilities for translation of tumor and transplantation immunology to the clinic. Additionally, there are world-class investigators that make up the Internal Advisory Committee and External Advisory Board. The focus of IM-TAG is significantly underrepresented in training programs leading to a paucity of investigators working in these areas. IM-TAG supports the training of two postdoctoral fellows yearly. There are 11 Professors, and eight Associate Professors, with outstanding qualifications who serve as mentors. We have identified two exceptional Assistant Professors, who serve as co-mentors. These investigators’ interests include 1) tumor vaccine design and vector generation, 2) adoptive cellular therapy, 3) the role of immune cells in the tumor microenvironment (TME), 4) immunogenomic assessment of TME, 5) enhancing our understanding of the biology of the graft-versus-tumor response and 6) approaches to manage acute and chronic graft-versus-host disease. There are innovative mechanisms for training that include 1) didactic coursework in statistical design, 2) immunogenomic coursework and data evaluation overseen by world experts in tumor immunogenomics , and 3) participation in Immunotherapy Working Groups, which are translational teams that assist in the development of novel immunotherapies, allowing trainees to be involved in the development of INDs and clinical trials. We have a comprehensive approach to train highly meritorious individuals pursuing postdoctoral studies, in addition to a summer course for undergraduate students to provide them with hands-on experience in immunotherapy or transplantation immunology. Strengths of the program include 1) an extremely well-funded group of mentors with substantial experience training postdoctoral fellows, 2) an outstanding leadership group with a long history of leading collaborative research proposals, 3) and the presence of a world-class vaccine and transplantation infrastructure that trainees can access. This program combines the mentors, foundation, and resources of a nationally recognized tumor immunology, cancer immunotherapy and transplantation immunology program with novel approaches to enhance vertical training and enhance the bench to bedside development of therapeutics by individuals who are trained in this program.