Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2026 · 2025-03

SUMMARY Genetic mutations that alter UBE3A ubiquitin ligase activity are linked to several neurodevelopmental disorders. Angelman syndrome (AS) is a severe syndromic neurodevelopmental disorder that is caused by deletion or loss- of-function (LOF) mutation of the maternally inherited copy of UBE3A. In contrast, duplication of maternal or paternal UBE3A is associated with Dup15q syndrome, one of the most common forms of autism spectrum disorder. And gain-of-function (GOF) missense mutations in UBE3A cause neurodevelopmental phenotypes that are distinct from AS in humans and in mouse models. Therapeutics are being developed to increase or decrease UBE3A as treatments for AS and Dup15q, respectively. While reporter assays exist to evaluate UBE3A activity, these assays cannot be used to measure endogenous UBE3A activity in cells or in the brain non-invasively over time. This substantially limits the speed at which therapeutics that normalize UBE3A levels in vivo can be tested and advanced for these debilitating disorders. In preliminary studies, we developed a highly sensitive luciferase- based biosensor that can quantify endogenous UBE3A activity in cells and primary neurons. This biosensor can also quantify the spectrum of UBE3A activity associated with UBE3A LOF and GOF missense mutations. Here, we made additional modifications to the biosensor to improve its sensitivity to changes in UBE3A activity. We hypothesize that this biosensor with improved sensitivity can be used to quantify aberrant UBE3A activity in cells and in the brain of live animals that model various forms of autism. We propose to evaluate the efficacy of this UBE3A biosensor in primary cells (neurons, fibroblasts, blood cells) and in the brain of mouse lines that model AS (maternal Ube3a deficiency), Ube3a gene duplication, and an autism-linked Ube3a GOF mutation. We will also generate and validate a knock-in mouse harboring this UBE3A biosensor expressed from a neuron-specific promoter. This UBE3A biosensor knock-in mouse line could be used in future studies to non- invasively monitor the efficacy and durability of therapeutics that increase or decrease UBE3A activity, as respective treatments for AS and various forms of autism.

NIH Research Projects · FY 2026 · 2025-03

The first two years are an exceptionally dynamic and critical period of brain development, featuring significant growth in both cerebrum and cerebellum. The availability of large-scale, multi-site infant MRI datasets, e.g., the developing Human Connectome Project (dHCP), Baby Connectome Project (BCP), and National Database for Autism Research (NDAR), affords unprecedented opportunities for precise charting the dynamic early brain development, providing important insights into the origins and aberrant growth trajectories of neurodevelopmental disorders, such as autism. However, existing neuroimage analysis tools designed for adults are not suitable for infant neuroimages. In 2020, our team has successfully developed and released iBEAT V2.0 (infant Brain Extraction and Analysis Toolbox) with advanced deep learning techniques, which has successfully processed 31,000+ infant scans from 200+ institutions with diverse imaging protocols and scanners. iBEAT has directly contributed to 50+ journal publications, including Brain, Nature Methods, Neuron, Nature Communications, PNAS, Neuroimage, and Cell Reports. However, iBEAT still has two major limitations. 1) It focused on the infant cerebrum MRIs and thus is inapplicable for the more challenging cerebellum MRIs, which exhibit much thinner and more tightly folded cortex than the cerebrum, extremely low and dynamic tissue contrast, and suffer from large domain-shift issue across imaging sites. 2) Certain important functionalities for cerebrums are either missing, e.g., motion correction, subcortical segmentation, volumetric parcellation, and surface registration, or have degraded performance in certain scenarios. To address these issues, this project aims to significantly enrich iBEAT by 1) creating deep learning-based computational tools for cerebellar tissue segmentation, atlas building, surface reconstruction and parcellation, as all as 2) adding new cerebrum-related functionalities and significantly improving existing functionalities, to enable comprehensive, accurate, and integrative analysis of cerebrum and cerebellum and their interplay during infancy. Accordingly, we propose five specific aims. Specifically, we will develop a novel prior-guided cerebellum tissue segmentation method with self-verification (Aim 1). We will then construct the first 4D infant cerebellum atlases with longitudinally consistent, temporally continuous, and spatially detailed patterns, by developing a novel unsupervised learning-based anatomy-guided atlas construction framework (Aim 2). We will reconstruct topologically correct and geometrically accurate cerebellar cortical surfaces and further develop a novel Spherical Surface Transformer to precisely parcellate cerebellar cortical surfaces into anatomically meaningful regions (Aim 3). We will add new cerebrum-related modules for motion correction, subcortical segmentation, volumetric parcellation, and surface registration, and further improve existing modules in terms of robustness and accuracy with our new techniques (Aim 4). Finally, we will undertake a comprehensive upgrade for iBEAT, by improving usability, robustness, compatibility, code structure, and documentation (Aim 5).

NIH Research Projects · FY 2026 · 2025-03

ABSTRACT In 2023, there will be an estimated 81,180 new cases of bladder cancer in the United States, resulting in an estimated 17,100 deaths. Over 90% of bladder cancers are urothelial carcinomas (UC) and the mortality associated with metastatic disease remains particularly grim. APOBEC3 (apolipoprotein B mRNA editing enzyme catalytic polypeptide like) is a family of enzymes that catalyze the deamination of cytosine nucleotides and has a physiologic role in both B cell receptor diversification as well as antiviral defense by restricting viral replication through cytidine deamination of the viral genome. There are 7 APOBEC3 family members in human and 1 Apobec3 in mouse. Aberrant expression of APOBEC3A (A3A) and APOBEC3B (A3B) has been shown to be upregulated in many cancers, but the APOBEC mutational signature is most highly enriched in bladder cancer. Indeed, nearly 70% of single nucleotide variants (SNVs) in bladder cancer are attributable to APOBEC induced mutagenesis. To address the role of APOBEC3 enzymes on bladder cancer initiation and progression, we generated a novel mouse strain: Rosa26LSL-Apobec3 and crossed them with our previously reported UPP mice that inactivate Trp53 and Pten in Upk3a expressing intermediate/umbrella cells through Cre-mediated recombination. These UPPA mice develop tumors with shorter latency than control UPP mice and have a striking phenotype of squamous differentiation. Our preliminary work has shown that Apobec3 is sufficient to promote squamous differentiation and that IL1A is both necessary and sufficient for squamous transdifferentiation of bladder cancer cell lines and normal urothelial organoids. Analysis of primary bladder tumors demonstrates that human APOBEC3A and IL1A, but not APOBEC3B, is associated with squamous differentiation in human bladder tumors. Moreover, APOBEC3A is significantly upregulated in chemotherapy resistant tumors. Lineage plasticity is a known mechanism of treatment resistance and squamous transdifferentiation has been recognized as an important manifestation of lineage plasticity and as a documented mechanism of therapy resistance to kinase inhibitors in lung adenocarcinomas as well as KRAS G12C inhibitors. Successful completion of this proposal will give us a better understanding of APOBEC3’s role in bladder tumorigenesis and in particular an in depth understanding of how it promotes squamous differentiation, to what extent APOBEC3 mediates resistance to currently approved therapy for bladder cancer, and whether co-inhibition of the IL1A pathway can be leveraged in the future for reversal of APOBEC-induced treatment resistance.

NIH Research Projects · FY 2025 · 2025-02

PROJECT SUMMARY / ABSTRACT Microbiomes have been described as ‘ecosystems on a leash,’ because immunological control of the microbiome is critical for host health. Variation in the genes that produce the immune system has been linked to microbiome dysbiosis and to disease, and studying immune systems in the context of host-microbe coevolution has generated critical insights. However, we do not understand the evolutionary forces maintaining host genetic variation related to the microbiome or how immune systems coevolve with beneficial microbes. My research program takes advantage of several key features of an innovative animal model system: an insect called the pea aphid. This animal reproduces asexually, and we can precisely manipulate bacterial microbiomes across asexual lineages by adding or removing specific strains and species of microbes. The aphid microbiome includes species of Enterobacteriaceae that have clear benefits for their hosts including protection against pathogens and parasites. These aspects of our model system allow us to untangle the specific effects of host and bacterial genetic variation on the microbiome, and to study natural and ecologically- relevant host-microbe pairings. Importantly, my lab has generated key data on the patterns of association between microbes and host populations, and we have shown that genetic variation in the immune system governs the association with bacterial symbionts. Over the next five years, we will use techniques adapted from human innate immunology, functional and forward genetics, and dual-RNAseq of both host and microbes to understand the roles by which animal immune systems and beneficial microbes evolve in a bi-directional manner. Our preliminary data suggest that phagocytes control the density of bacterial infections, but that some strains of microbes have evolved to kill phagocytes to grow to high density. We will test a mechanistic model for the interaction between phagocytes and beneficial bacterial in the context of host and bacterial genetic variation. Simultaneously, we will use an F2 cross to determine what kinds of genes underlie host genetic variation in the microbiome, the patterns of molecular evolution at these loci, and importantly, whether these genes also govern resistance against often closely-related pathogens. Together, this work will address fundamental questions relevant to human health, including whether adaptation to beneficial microbes trades off with the ability to combat pathogens, and how control of the microbiome shapes microbial virulence. This work will establish a foundation for my lab’s future efforts to develop a comprehensive picture of how natural selection shapes host traits related to the microbiome in a controlled and tractable model system.

NIH Research Projects · FY 2026 · 2025-02

ABSTRACT Air leak after lung surgery, especially prolonged air leak (PAL) (≥ 6 days) is the bane of existence for thoracic surgeons and patients. Air leaks arise from 3 sources after lung surgery: through a staple line placed across lung tissue, as part of lung resection, air leaking from a lung surface opened to separate incomplete fissures to remove a lobe; and Injury to lung tissue unrelated to a lung resection. Air leak after lung surgery requires a chest tube, often prolongs hospitalization, increases cost and death rate, and decreases quality of life (QOL). There are strategies to decrease air leak after lung surgeries, an FDA-approved sealant, and strategies to discharge patients home with a chest tube. We propose a way to eliminate air leaks after lung resection. This would be the most cost-effective way to facilitate early ambulation, reduce pain, prevent chest tubes, and improve QOL after lung resection. We developed anisotropic and auxetic hydrogel patches (PAAx) that can be placed adherent to the lung surface and stretch with lung movement. Therapeutics can be applied to lung surfaces on both sides of these patches. We showed in rats that air leaks from ventilated perforated lung tissue can be sealed by application of fibrinogen to the side of the PAAx applied to the lung, then thrombin applied on the opposite side of the patch. This combination allows fibrin to seal the holes in the lung and helps hold the patch in place. This results in prevention of air leak under positive pressure ventilation, even if tidal volume (TV) is increased to 15 ml/kg. We showed similar results in porcine lungs in an ex-vivo model. Air leak after perforation with an 18-gauge needle was stopped by application of a PAAx treated with fibrinogen and thrombin (F&T), even with inflation to TV of 15 ml/kg. We will test the hypothesis that these PAAx when applied to the surface of lungs in rats with perforations and lung resections, and in pigs after multiple wedge resections and non-anatomic lung resections will have air leaks stopped so that a chest tube will not be required after lung surgery. This hypothesis will be tested by accomplishing these specific aims: Aim 1: To optimize patch degradation rate and elastic properties using in vitro studies. Aim 2: To determine if application of fibrinogen and thrombin (F&T) to PAAx patches will prevent pneumothorax (PTX) compared to F&T alone using survival rat lung injury models. PTX will be quantified by CT scan 3 and 10 days after lung injury, then injured lung areas will undergo histologic examination to assess inflammation from the patch. Aim 3: To determine if PAAx/F&T patches can prevent air leak in non-survival large animal (porcine) lung models from staple lines after wedge resection, by treating staple lines, and open lung surfaces in live anesthetized pigs. Many patients have air leaks after lung resection. Some are discharged home with chest drainage systems. If any air leak following lung surgery could be consistently eliminated, chest tubes would be unnecessary, complications and cost could be substantially reduced, and QOL improved for patients having lung surgery. These novel hydrogel patches with FDA-approved therapeutics (F&T) may provide an innovative solution to a vexing clinical problem.

NIH Research Projects · FY 2026 · 2025-02

Project Summary Critical questions remain regarding the recovery of lung structure and function following acute respiratory distress syndrome (ARDS) and pneumonia. A subset of immune cells, Foxp3+ regulatory T lymphocytes (Tregs), are essential in resolving inflammation for several experimental models of acute lung injury (ALI). Furthermore, several groups, including ours, have established that Tregs are present in the lungs of patients with ARDS, suggesting Tregs contribute to recovery during ARDS. While therapies are limited for ARDS, several trials have illustrated the mortality benefits of glucocorticoid (GC) therapy for ARDS and pneumonia. The mechanistic effects of GC are pleiotropic, and their influences on Treg phenotype and function are varied and remain ill- defined during ARDS. Furthermore, the heterogeneity of patient responses to GC and the contribution of the etiology and severity of the illnesses, the timing of therapy, and the dosage of GC to GC's efficacy all merit more investigation. Our published studies in murine ALI demonstrate that during the resolving phase of lung injury, Tregs expand in number and change their gene expression profiles compared to Tregs in uninjured control lungs. Treg transcriptome profiles suggest changes in gene expression that may contribute to Treg-driven lung repair. One transcript upregulated in Tregs isolated from resolving lung tissue is the glucocorticoid receptor (GR) (Nr3c1). Our Aims seek to test the hypothesis that glucocorticoid signaling occurs through Tregs, and this Treg- signaling is critical to GC effects. Aim 1 proposes to determine the function of Treg’s GR signaling and the direct and indirect impact of GCs on Treg-promoted resolution. We hypothesize that the administration of exogenous GC affects Treg responses during resolution. Additionally, we will test the hypothesis that GR-deficient Tregs have decreased immunomodulatory and tissue reparative effects during ALI. Aim 2 addresses the effects of host genetic variations on the functions of Tregs and the effects of GCs. Host genetic variation likely impacts on the immunologic response during ARDS and are determinants of outcomes. The Collaborative Cross (CC) is a multi- parental genetic reference population generated using eight founder mouse strains. These genetically diverse CC strains demonstrate phenotypic variability useful for genetic mapping disease trait-associated quantitative trait loci (QTL). Variation in Treg phenotypes in the CC has been reported. Aim 2 proposes leveraging the CC's genetic diversity to test the hypotheses that 1) host genetic variation impacts the response to exogenous GC administration after inducing ALI and 2) variation in the Treg number or effector phenotypes across the CC associates with different rates of resolution of ALI and identifies genetic loci that define genes of host variation. We predict that the proposed studies will identify genetic loci that modulate Treg responses critical in resolving ALI and will identify the role of GCs in promoting resolution. Identifying and elucidating these mechanisms of GC action and GR function may identify possible therapeutic approaches to lessen collateral tissue damage without negatively altering the response to injury.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Developing innovative solutions to prevent disease onset in individuals at high-risk for knee osteoarthritis (KOA) is paramount to advancing overall societal health and promoting the economic stability of our healthcare system. Aberrant limb-level loading during walking is hypothesized to be a determinant of disease onset in young individuals at high-risk of KOA following knee injury, thus positioning precision gait rehabilitation as a critical future intervention to reestablish limb-loading strategies to mitigate KOA onset. However, the current evidence linking aberrant gait biomechanics with KOA development is primarily observational and lacks the fundamental scientific rigor of an established mechanistic pathway to explain how limb-level loading alters mechanical, biophysical, and biological properties of tibiofemoral articular cartilage. Identifying the underlying mechanistic pathway linking limb-level loading to articular cartilage changes is the single most important scientific knowledge gap that needs to be overcome to advance precision gait retraining as a strategy for KOA prevention. Our overarching hypothesis is that a sustained compressive limb-level loading biomechanical gait phenotype, which is found in individuals at high risk for KOA following knee injury, leads to aberrant sustained tibiofemoral articular contact forces (i.e., mechanical), causing greater cartilage strain (i.e., biophysical) and more deleterious changes to cartilage composition and joint tissue metabolism (i.e., biological), thereby contributing to KOA development. We hypothesize that directly modifying the sustained limb-level loading profile with dynamic limb-level loading will reverse these deleterious mechanical, biophysical, and biological cartilage changes. The proposed R01 will overcome this scientific gap and test our overarching hypothesis: i) in vivo using real-time gait biofeedback to precisely adjust limb-level loading in participants with anterior cruciate ligament reconstruction (ACLR) and determine resultant acute biomechanical and biophysical tissue changes and the cumulative biological cartilage changes caused by sustained and dynamic limb-level loading conditions; and ii) ex vivo using human articular cartilage explants to directly apply phenotypic loading profiles to the tissue, establishing a critical cause-and- effect understanding of sustained and dynamic loading on tissue mechanics, biology, and histology. Completion of our three specific aims will: 1) determine the acute in vivo mechanistic link between sustained and dynamic limb-level loading on mechanical and biophysical tibiofemoral articular cartilage outcomes; 2) determine the cumulative in vivo mechanistic link between sustained and dynamic limb-level loading and biological tibiofemoral articular cartilage outcomes; and 3) use an ex vivo experiment in human cadaveric tissue to establish the cause- and-effect relation between phenotypic (sustained vs. dynamic) loading and biological, biophysical, and histological changes in articular cartilage explants. Our R01 is innovative, as it is the first to study a mechanistic pathway defining the link between sustained limb-level loading and outcomes associated with KOA development and significant as the knowledge gained will directly inform development of future precision gait rehabilitation.

NIH Research Projects · FY 2026 · 2025-01

ABSTRACT Lipoprotein(a) [Lp(a)], a highly atherogenic, prothrombotic, and proinflammatory lipoprotein, is a causal, independent, and highly heritable (h2=70-90%) atherosclerotic cardiovascular disease (ASCVD) risk factor that is elevated in 1.5 billion people globally. Despite significant stakes and emerging therapies that can reduce Lp(a) by >80%, few studies have comprehensively identified and characterized the potentially broad phenotypic effects of Lp(a). This major research gap overlooks opportunities to anticipate adverse effects of therapeutic Lp(a) lowering, illuminate mechanisms of action, and identify novel treatment indications. A second major research gap is the Eurocentric evidence base of Lp(a), which has persisted despite Lp(a) being recognized as one of the most variable ASCVD risk factors across populations. This research gap constrains the generalizability, relevance, and reach of evidence that informs Lp(a) clinical, regulatory, and public health decision making. A common factor underlying both research gaps is the absence of validated Lp(a) measures in broad studies with dense phenotypic data. We propose to address this obstacle by assembling a large and broad consortium with validated Lp(a) measures and genotypic data. To expand our consortium, we will measure Lp(a) in African, Polynesian, and South American cohorts using validated assays. Next, to facilitate causal inference in studies without measured Lp(a) but with dense phenotypic data, we will leverage our consortium and statistical genetics advances to construct highly accurate Lp(a) polygenic risk scores (PRS) in all populations. These PRS will then be projected into biobanks with dense genotypic and phenotypic data, but no Lp(a) measures. Finally, we propose a suite of causal inference studies that substitute Lp(a) PRS for measured Lp(a) and examine broad phenotypes. These studies enable well-powered (n=1,137,708), comprehensive, and generalizable causal inference investigations that are robust to confounding and reverse causation and examine broad phenotype classes. By identifying and characterizing both the anticipated and unanticipated effects of Lp(a), the proposed study will provide an essential foundation for future efforts that aim to maximize the benefits and minimize the risks of therapeutic Lp(a) lowering for everyone.

NIH Research Projects · FY 2026 · 2025-01

Project Summary The proposed research seeks to validate and extend a platform technology for the delivery of protein-based drugs, which includes application to current and next generation therapeutics. The bioengineering strategy employs the designed assembly of endogenous biomolecules, namely thrombolytic enzymes and vitamin B12, as well as cell-based transport and delivery to create a biocompatible drug delivery platform. The drug delivery technology uses the circulatory system as a natural drug depot, is drug agnostic, releases the protein therapeutic at the diseased site in a photon-targeted fashion, and is 25-fold more effective than the standard of care in our preliminary in vivo studies. The wavelength of drug release is readily assigned at the molecular level. The following issues will be addressed: (i) accommodation of an array of thrombolytic proteins that operate by distinct mechanisms; (ii) multi-drug synergism of thrombolysis (iii) responsiveness to deep tissue penetrating photons; (iv) delivery efficacy as a function of irradiation parameters, melanin and body mass index, and sex; and (v) therapeutic efficacy and safety under conditions in which therapy is contraindicated.

NIH Research Projects · FY 2026 · 2025-01

Vibrio cholerae causes the waterborne diarrheal disease, cholera, with annual fatality rates reaching 120,000 worldwide. Cholera outbreaks are common after natural disasters including earthquakes, storms, and floods where a reservoir of V. cholerae persists as aquatic biofilms (surface-attached microbial communities that are composed of microorganisms and a matrix composed of extra-polymeric substances, such as exopolysaccharides, proteins, and nucleic acids) found in sewage contaminated waters. Biofilms are complex structures consisting of cells which have distinct functions. The goal of this project is to identify the cellular states occupied by different V. cholerae cell populations within biofilms. We will determine how different cell types organize in 3-dimentional space during biofilm structure formation and how specific cells bring about the dispersal of biofilm in response to host signals present at the onset of infection. We will first measure the single cell transcriptome of thousands of single V. cholerae cells grown in laboratory culture media in either planktonic state or a biofilm using a microfluidic bacterial single- cell RNAseq assay we recently developed. We will compare the level of cell-to-cell heterogeneity in biofilm vs planktonic state at different times of growth. This data will identify the physiological states in which bacteria exist within the biofilm. After examination of the inherent cell-cell heterogeneity within biofilms we will use a fluorescence microscopy approach to determine the spatial arrangement of each cell-type within the 3-dimentional structure of biofilms. During V. cholerae colonization of the human GI tract ingested biofilms disperse into single cells upon contact with host-secreted bile acids. After we characterize the early formation, organization and maturation of biofilms we will next determine which cellular changes occur during biofilm dispersal in a model that utilizes specific bile-acids and temperature for biofilm dispersal. Our data will provide the first transcriptome-wide measurements of the transcriptional plasticity of V. cholerae in biofilms, their natural infective reservoir, and will be useful in identifying states associated with the onset of infection.

NIH Research Projects · FY 2025 · 2025-01

Abstract The global burden of Alzheimer’s Disease (AD) and related dementias (ADRD) is projected to rise dramatically in the coming years. Memory is a key marker of cognitive deficits but its measurement in global studies of population aging is difficult. Word-based measurements of memory are challenging to interpret in populations where literacy is low, literacy inequality is high or multiple languages are in common use. These concerns are likely less important for olfactory-based assessments based on neurobiology. The entorhinal cortex is a key center for olfactory memory, linking the hippocampus with the olfactory bulb, and is among the first areas affected by AD neuropathological hallmarks, particularly neurofibrillary tangles. This project will rigorously evaluate the value-added of an olfactory-based approach to measuring memory function and adapt the protocol to facilitate its inclusion in population-based surveys across the globe, which will support production of comparable measurements across contexts. The Percepts of Odor Episodic Memory (POEM) is a recently developed protocol that measures olfactory memory and has been shown to be predictive of ADRD among older adults. We are fielding POEM in the Study of the Tsunami Aftermath and Recovery (STAR), a population- representative longitudinal survey of respondents who were living along the coast of Aceh, Indonesia, at the time of the 2004 Indian Ocean tsunami. We also collect a comprehensive battery of word- and drawing-based memory assessments as part of the Harmonized Cognitive Assessment Protocol (HCAP). Leveraging the design of STAR, and the fact that among those living in similar coastal communities, exposure to the tsunami can be treated as random, this project will provide new evidence on the long-term effects of exposure to the stresses of the disaster on memory function and cognitive aging. We will investigate the relationships among the array of memory measures and each of their relationships with age, sex and education as well as with exposure to the stressors of the tsunami. To place these results in context, we will analyze HCAP data from the U.S., England, India and Mexico to contrast the relationships with socioeconomic and demographic characteristics across places with different levels of literacy and language heterogeneity in an effort to identify generalizable findings within and across settings. Finally, we will field an Evaluation and Adaption Study (EASy) of POEM to develop a protocol that can be fielded at low cost in any field setting and provide measures of memory that are comparable across settings. EASy will evaluate how variation in assessment length and the number, order, type, range and familiarity of scents affects measurement in order to reduce the time-burden of the assessments and build a well-documented, ready-to-adopt protocol that is simple to implement in other field settings. Together, these data will afford the development of a robust olfactory memory test that can deployed across variable global populations at risk for ADRD.

NIH Research Projects · FY 2026 · 2025-01

ABSTRACT Over the next decade, the number of lung cancer survivors is expected to rise considerably in the United States, due to an aging and growing population and a shift toward more early-stage lung cancers detected from increased lung cancer screening uptake. After treatment with curative-intent surgery, however, many survivors of early-stage non-small cell lung cancer (NSCLC) remain at high risk for lung cancer recurrence or second primary lung cancer (SPLC). To detect recurrence and SPLC at their earliest stages, postoperative imaging surveillance with chest computed tomography (CT) is generally recommended every six months for the first two years and annually thereafter. Yet, this clinical recommendation is based on limited evidence of benefits and harms, with no consideration for individual differences in risk of recurrence or SPLC. Routine chest CT images are a potentially useful, but largely untapped, data source for predicting risk of recurrence or SPLC to tailor surveillance among lung cancer survivors. Chest CT scans contain specific imaging biomarkers of body composition and cardiopulmonary health, as well as non-specific imaging data that are amenable to analysis using deep learning, all ascertainable without additional intervention, risks, or costs to survivors. Studies including ours suggest that skeletal muscle and adipose tissue measured from CT scans can predict outcomes after NSCLC resection. In the context of lung cancer screening, we have also developed Sybil, a validated deep- learning model that uses information from a single low-dose chest CT scan, to accurately predict future incident lung cancer risk. Our overarching goal is to optimize survivorship of early-stage NSCLC following curative-intent surgery by incorporating a risk-based surveillance strategy that leverages routinely available imaging data. Our multidisciplinary team proposes to examine 12,000 individuals treated surgically for stage I or II NSCLC from 2015 to 2025, with follow-up for outcomes through 2027, using longitudinal electronic health records and serial chest CT scans from three healthcare systems that serve distinct and diverse populations. First, we will determine real-world practice patterns and effectiveness of imaging surveillance, overall and by individual characteristics. Second, we will develop and validate risk prediction models for lung cancer recurrence and SPLC that incorporate imaging biomarkers of body composition and of vascular and pulmonary health derived from preoperative and postoperative surveillance CT scans. Lastly, we will assess and then optimize Sybil’s performance in predicting lung cancer recurrence or SPLC using postoperative surveillance CT scans. Overall, the proposed study will advance knowledge about the promising potential for personalized risk-stratified surveillance of recurrent or new disease using novel imaging-driven methods among the growing population of lung cancer survivors.

NIH Research Projects · FY 2025 · 2025-01

PROJECT SUMMARY Sexual and gender minority (SGM) youth are at higher risk for problematic substance use compared to non- SGM youth. This disparity persists into adulthood, when SGM adults experience a disproportionately higher rate of substance use and disorder. Though such disparities are well established, few studies have examined social determinants that may explain variation in substance use risk within SGM youth. Minority Stress Theory suggests that variation in risk for substance use is due to individual differences in exposure to minority stress (e.g., discrimination). Social safety (feelings of social inclusion and acceptance) may be a key mechanism underlying the link between minority stress with substance use. Extant theory and research suggest that heightened experiences of minority stress have a negative impact on social safety, indicated by chronic threat vigilance. The proposed study seeks to investigate social safety as a mechanism explaining the heightened rates of problematic substance use among SGM youth. Furthermore, this study investigates the impact of protective factors that may disrupt this pathway and reduce substance use risk among SGM youth. Identification of social safety mechanisms, as well as associated protective factors, can inform the development of school-based programs, social services, and policies designed to specifically address social safety as a protective factor against risk for substance use among SGM youth. With this NRSA F31 proposal, the applicant seeks training to pursue a career as an independent clinical research scientist focused on prevention of problematic substance use among SGM youth. Training goals address gaps in her current doctoral program and include acquiring knowledge about theories, mechanisms, and methods for studying developmental pathways for youth substance use and SGM disparities more specifically; expertise in advanced longitudinal and integrative data analysis; and skills related to scientific writing and dissemination. These skills will be applied to conduct a study examining within-SGM population heterogeneity and timing of risk for substance use during youth development. Three aims will be investigated through integrated data analysis of two seven-wave studies of SGM youth using an accelerated longitudinal cohort design to capture development from ages 16 to 24. Data will be harmonized and pooled to meet psychometric standards before analyses. Specific aims are to examine whether: 1) SGM youth with greater minority stress in adolescence show greater decrements in social safety and, in turn, greater increments in problematic substance use into young adulthood as compared to those with less minority stress, 2) for any given SGM youth, individual periods of increased problematic substance use coincide with periods of greater minority stress and decrements in social safety, and, 3) protective factors reduce risk of problematic substance use associated with decrements in social safety across time and in period-specific elevations among SGM youth.

NIH Research Projects · FY 2025 · 2025-01

Abstract Summary A wealth of research shows that teams comprised of members with a diversity of backgrounds, identities and perspectives are more successful and creative in multiple domains. To recruit and retain such a biomedical workforce, the NIH created the F99/K00 mechanism to support talented graduate students through the transition into postdoctoral training and prepare them for careers in academia and non-academic sectors. The UNC Advancing Research Careers (ARC) Training Hub (UNC-ARC) is designed to provide cohort-based and individualized skills and career development and mentor training to assigned F99/K00 scholars, preparing them to be the next generation of leaders in biomedical science. UNC-ARC is a partnership between the UNC Office of Postdoctoral Affairs (OPA) and the UNC School of Medicine Office of Graduate Education (OGE) and will provide career-focused support to predoctoral (F99) and postdoctoral (K00) scholars. The training plan builds on the expertise of the OGE and OPA to deliver evidence-based professional development and mentor training to trainees and mentors. Both offices are also experienced in program evaluation, data tracking and education scholarship. We propose a hybrid program of virtual workshops and cohort meetings throughout the year, culminating in an annual, in person Summer Institute on the UNC campus. Key aspects of the program include: 1) the use of individual development plans to tailor professional development training and career preparation to each scholar’s career goals; 2) a core professional development curriculum delivered in a cohort-based format; 3) the assignment of external career mentors from work sectors aligned with the scholar’s career goals; 4) a robust suite of skills and career exploration training opportunities tailored to individual scholar’s career stage and goals; and 5) a range of networking activities designed to build strong mentor networks – including peer mentors – for each trainee. Trainee experiences and outcomes will be rigorously evaluated and disseminated to discover and promote best practices in graduate and postgraduate biomedical training. As a national leader in biomedical science education, UNC is well positioned to provide robust career support to F99/K00 scholars from across the United States through the UNC-ARC.

NIH Research Projects · FY 2026 · 2025-01

PROJECT ABSTRACT Studies of people infected with HIV-1 from the subtype B lineage of virus have led to important insights on the nature of the transmitted virus, early dispersal of the virus throughout the body, evolution of macrophage-tropic virus late in the course of infection (especially in the CNS), and the phenomenon of latency (especially within CD4+ T cells in the lymphoid system). However, these questions have been studied to only a limited extent in people who have HIV-1 infections from the most common subtype, i.e. subtype C. In addition, the nature of viral evolution within the CNS to form macrophage-tropic HIV-1, and the cell types that may contribute to a latent reservoir within the CNS are poorly understood. The proposed work is built on the availability of a unique autopsy tissue repository and a novel model cell culture system to explore these questions in the context of infection with HIV-1 from subtype C lineage. The tissue repository for autopsy tissue from recently deceased individuals infected with subtype C lineage has been established at the University of the Witwatersrand by Drs. Papathanasopoulos and Wagner, and additional autopsy tissue samples are being collected for this repository. In addition, Drs. Symons and Nijhuis have access to viable human brain tissue (from non-HIV donors) collected during rapid autopsy which they have used to develop in vitro infection models for primary microglia and single nuclei analysis. These tools, coupled with Drs. Swanstrom and Joseph expertise in viral evolution and genetics are the basis of an international collaboration that will bring new insights into the nature of HIV-1 latency in the brain and do so in the setting of the most common lineage of HIV-1 infection. With these aims we will: i) determine the frequency that macrophage-tropic virus evolves within the immune-privileged confines of the CNS; ii) identify which cell types within the CNS are expressing virus; iii) determine how early macrophage-tropic virus evolve in the disease course; iv) compare long-lived viral DNA populations in the CNS to those in lymphoid tissue to determine if there are compartmentalized reservoirs; v) sort nuclei based on cell origin to determine which CNS cell types contain long-lived viral DNA; vi) use sorted nuclei to search for virus-expressing cells; vii) use M-tropic subtype C viral clones to establish a validated model of viral latency in primary microglia cells. Finally, we will explore these questions in samples collected from both men and women to examine if sex is a significant biological determinant of the nature of the CNS reservoir.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Challenging or restrictive eating behaviors are well-documented in autistic individuals. Restrictive eating behaviors, such as food selectivity, refusal and neophobia occur at a greatly increased rate in autism. Eating disorders, such as anorexia nervosa, avoidant restrictive food intake disorder, and binge eating disorder, are also diagnosed at highly elevated rates in autism. However, the associations between eating behavior profiles and later clinical outcomes, such as eating disorder diagnoses, are poorly understood, with a lack of longitudinal data to meaningfully capture these trajectories. This is an urgent research need given the clinical significance of restrictive eating at a nutritional, behavioral, and social level. Given the historic sex difference in autism diagnoses, most studies fail to fully consider the role of sex on eating behaviors. However, in neurotypical populations, there are variations in challenging eating behaviors and rates of eating disorders by sex. In more recent studies, these trends have been mirrored in autistic samples The overarching goal of this NIMH R03 is to leverage data across four existing cohorts of autistic and non-autistic individuals, enriched for females and spanning a wide age range (4 to 39 years), to identify profiles of eating behaviors in autism and determine how these vary by age, and sex. Importantly, these studies all employed harmonized measures of eating behaviors to achieve a total sample size of approximately 1,200 individuals. This R03 has two aims: (1) Using latent profile analysis (LPA), characterize profiles of eating behaviors in a large sample of autistic and non-autistic individuals; and (2) Determine how profiles of eating behaviors vary by diagnosis, age, and sex. This R03 directly addresses NIMH Goal 2 to examine mental illness trajectories across the lifespan studying eating behaviors across a wide age range, spanning early childhood through to adulthood, and pan-NIH goals, including studying sex as a biological variable, and leverages samples that over-sampled for autistic females. This data driven approach will enable us to identify age-related trends in challenging eating behaviors in autism. Such data will allow us to generate hypotheses that can be tested prospectively within our ongoing NIH-funded longitudinal samples, spanning early childhood through to young adulthood, to understand the mechanisms by which challenging eating behaviors escalate to an eating disorder.

NIH Research Projects · FY 2026 · 2025-01

Project Summary Long-term exposure to ambient air pollutant ozone (O3) is associated with decreased lung function and the development and/or progression of asthma and chronic obstructive pulmonary disease (COPD). The underlying mechanisms remain obscure, however. We seek to identify new mechanisms by which O3 exposure causes these common, chronic diseases by focusing on a paradoxical yet highly reproducible finding in human controlled O3 exposure studies: while a single O3 exposure causes airway inflammation, injury, and decreased lung function, responses decrease–rather than increase–after repeated O3 exposures, a process referred to as adaptation. Intriguingly, not all individuals adapt, leading to our overarching hypothesis that failure to adapt (FtA) may render individuals susceptible to the development of asthma or COPD after long-term O3 exposure. Using a new mouse model of O3 adaptation involving four consecutive O3 exposures (“4X O3”), we ruled out previous hypotheses for mechanisms of adaptation, most prominently upregulation of antioxidants. Instead, our new data point to alveolar macrophages (AMs) as key determinants of adaptation. We found that 4X O3 exposures lead to an altered transcriptome in AMs, rendering them hypo-responsive. Further, using genetically diverse mice from the Collaborative Cross (CC) mouse genetics reference population, we identified mouse strains that adapt (A) or fail to adapt (FtA) after 4X O3 exposure, mimicking the diversity of responses observed in human studies. Comparing the genomes, epigenomes, and transcriptomes of AMs from these phenotypically divergent strains will enable us to identify novel genes and pathways (i.e., new mechanisms) that influence adaptation to O3. In Aim 1, we will test the hypothesis that genetically-encoded differences O3-induced gene expression in AMs underlie differences in adaptation. We will profile the transcriptomes and epigenomes of AMs in phenotypically divergent CC strains, revealing genes and pathways associated with adaptation. Through subsequent bioinformatic analyses, we will identify putative regulators of differential response, and then experimentally validate their roles in vivo using a lentivirus-based gene targeting approach. In Aim 2, we will identify genetic loci that that drive adaptation through the classical genetic approach of quantitative trait locus (QTL) mapping. We will identify candidate genes at QTL by integrating omic data generated in Aim 1, then functionally test these genes in vivo. Finally, in Aim 3, we will explicitly test the hypotheses that failure to adapt is a predictor of susceptibility to long-term, O3-induced chronic lung disease and that AMs are a critical to this process. We will compare the incidence and severity of chronic lung disease phenotypes caused by 5-week O3 exposure between CC-A vs. CC-FtA strains, and test whether modifying AM gene expression using a long-lived lentivirus approach modifies outcomes after 5-week O3 exposure. Completion of our aims will lead to the identification of genes and pathways that underlie adaptation and improve our understanding of susceptibility to air pollution-induced lung disease.

NIH Research Projects · FY 2026 · 2025-01

Low Oxygen Environments (LOEs) are a common characteristic of numerous physiologically relevant conditions in many different locations in the body. Accordingly, understanding how LOEs impact the cellular response to technologies that can investigate and manipulate cells is an essential area of fundamental research that can lay the foundation for advancing health. Recently, lipid nanoparticles (LNPs) have emerged as promising technologies that can investigate and manipulate cells using messenger RNA (mRNA). Studies investigating the impact of LOE on mRNA LNP are currently limited, and addressing this key gap in our knowledge will be essential for understanding how to best investigate and manipulate cells in LOE using mRNA LNPs. Here, we will address this key gap in knowledge by studying two overarching Research Areas – Mechanism (Research Area 1) and Tool Development (Research Area 2). In Mechanism, we will perform comparative analyses exploring how various states of LOE impact the cellular uptake mechanisms of mRNA LNPs across different cell types, mRNA sequences, and LNP types. In Tool Development, we will develop Low Oxygen Environment Nanoparticles (LOENRs), a novel class of tools for better investigating and manipulating cells under LOE. In undertaking this approach, we hope our research will provide key fundamental insights about how cells respond to LOE, a common characteristic of physiologically relevant conditions that arise in many different locations in the body.

NIH Research Projects · FY 2026 · 2025-01

Cardiomyocytes (CMs), the most prevalent cells in the adult heart, are responsible for driving cardiac contraction. During the neonatal stage, CMs undergo a constellation of molecular, structural, and functional changes known collectively as CM maturation to enhance their ability to generate efficient and forceful contraction throughout postnatal life1. CM maturation has received increased attention recently due to the relative immaturity of pluripotent stem cell-derived CM, which limits its application in regenerative medicine. The importance of understanding postnatal CM maturation is also highly relevant to understanding human heart diseases. Defining key factors and signaling pathways that regulate CM maturation will offer valuable guidance for promoting CM maturation and future diagnosis and treatment of heart disease patients. Protein arginine methyltransferases (PRMTs) are a family of enzymes that catalyze arginine methylation on targeted protein substrates. As one of the most abundant post-translational modifications, protein arginine methylation has been linked to the regulation of a wide range of biological processes. Yet, the roles of PRMTs in CM maturation remain largely unexplored. In this research program, we focus on coactivator-associated arginine methyltransferase 1 (CARM1), the founding member of PRMTs also known as PRMT4. We found that Carm1 mutant CMs exhibit multiple maturation defects, including reduced cell and myofibril size, perturbed mitochondrial fusion, disrupted T-tubule formation, and compromised electrophysiological maturation. Mechanistically, CARM1 regulates genes that underlie CM structural and electrophysiological maturation at both transcriptional and posttranscriptional levels, demonstrating a critical and multifaceted role of CARM1 in controlling CM maturation. We thus hypothesize that CARM1-mediated transcriptional and posttranscriptional regulation controls multiple aspects of CM maturation. Our study will fill critical gaps in our understanding of the transcriptional, epigenetic, and post-transcriptional regulation of CM maturation. This new knowledge, in turn, will undoubtedly pave the way for developing new strategies to augment CM maturation for regenerative medicine.

NIH Research Projects · FY 2026 · 2024-12

Abstract The NCATS Biomedical Data Translator (“Translator”) aims to augment human reasoning and accelerate scientific discovery through a federated system that integrates a broad range of biomedical data and knowledge, and reasons over them to answer translational science questions. During the Development phase (Phase II), the Translator program successfully implemented a system capable of answering certain types of clinical and translational questions. We propose advancements to make Translator an even more effective and compelling resource that will attract a broad and deep community of biomedical researchers. To achieve this transformation, we propose DOGSLED (Data, Ontologies, and Graphs to Support Learning and Enhance Discovery). DOGSLED will build on the best elements of the Phase II system—many of which were developed by members of our team—while improving breadth, integration, efficiency, explainability, usability, and sustainability. During Phase II of Translator, as members of the Ranking Agent, Exposures Provider, and Standards and Reference Implementation (SRI) teams, we worked with the Translator Consortium to build and integrate the ARAGORN Reasoning Agent, the ICEES Knowledge Provider, the Node Normalizer, and the Biolink Model. Building on that work, the DOGSLED team will collaborate with other proposed teams such as DOGSURF and ARAX-MGKG2, should they be awarded funding, to advance Translator to the next level, catalyzing user uptake and satisfaction. Our planned improvements center around performance, functionality, and transparency. Aim 1 (Create a Performant, Scalable, Reproducible Translator) involves improving reliability and performance by centralizing and unifying data ingest, data processing, and deployment in an integrated infrastructure component called BioPack. In addition to improving the efficiency of the system itself, this work will streamline and standardize the development process, reducing demands on future developers and making Translator more sustainable and extensible. To realize Aim 2 (Expand the Functionality of Translator), we will support new query types, leverage underutilized KPs, ingest or make better use of new and existing biomedical and clinical knowledge sources, and improve reasoning approaches. We will leverage large language models to enable users to add their own data in the form of publications and other text-based information as well as to query Translator using natural language. To achieve Aim 3 (Make Translator Fully Transparent to Users), we will track provenance at every stage, from initial data ingest all the way to ranked, evidence-supported answers to user queries. This will feed into improvements in answer scoring and will enable the system to provide better explanations to users. These advances will significantly expand the range of queries that users will be able to ask of the system, build confidence in the answers, improve system performance, and position Translator to keep pace with future developments in biomedical science. In concert with a multi-pronged user engagement and outreach strategy inspired by other successful consortia, the DOGSLED team will greatly expand Translator’s user base and help the program move toward its vision of Translator as a transformative scientific discovery tool used by a growing number of researchers.

NIH Research Projects · FY 2026 · 2024-12

ABSTRACT/SUMMARY Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are now thought to lie within a shared umbrella of neurodegenerative disorders, all characterized by neuronal loss, behavioral deficits, and ultimately death with no truly effective treatments. It is now generally well recognized that TDP-43 dysfunction (both loss and gain of function) represents the major disease hallmark that is linked the onset and progression of these devastating diseases. However, modeling ALS/FTD has been particularly challenging, in part since slightly too much or too little TDP-43 is toxic to neurons. Our goal for this R61/R33 proposal is to develop and optimize a new ALS/FTD animal model that more accurately recapitulates the human disease spectrum. This model is based on the unanticipated finding that TDP-43 can undergo acetylation on its lysine residues, and that this specific post-translational modification (PTM) has a dramatic effect on TDP-43; it disengages TDP-43 from its target RNAs, promotes its aggregation, and leads to the loss of normal nuclear TDP-43 function. Based on this mechanism, we developed a new CRISPR-integrated mutant mouse line that incorporated an acetylation- mimicking substitution (KàQ) at position K145 in the endogenous mouse TARDBP locus, thereby generating TDP-43K145Q mutant mice, and we observed many striking hallmarks of disease including aggregated TDP-43, widespread transcriptomic and splicing alterations, and cognitive decline. These mice should provide the ideal platform for biomarker therapeutic development opportunities, not just for sporadic ALS/FTD, but potentially any neurodegenerative disease within the TDP-43 umbrella characterized by abnormal TDP-43 deposition. The goal of this R33/R61 proposal is to extend and optimize the readouts to demonstrate a progressive ALS/FTD-like phenotype using aged mouse cohorts (R33 phase) and subsequently externally validate our animal findings with those in human models, including postmortem ALS/FTD tissues and iPSC-derived neurons (R61 phase). The use of iPSC-derived sporadic ALS “trio” lines, in which the lysine residue of interest (K145) is targeted, should provide insight into causality of TDP-43 acetylation as a driver of sporadic ALS/FTD. This study is ideal for the R61/R33 mechanism, since it will highlight TDP-43K145Q mice and newly developed iPSC lines as next-generation sporadic ALS/FTD models that could radically transform the field and provide new platforms for therapeutic testing of drug candidates.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY 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 CD4+ and CD8+ 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 T cell responses to infection in this high-risk population. T cell function and metabolism are closely linked, and many studies have demonstrated that changes to CD4+ and CD8+ T cell metabolism influence T cell fate and function. We have found that activated CD4+ T cells from obese mice have an altered metabolic profile characterized by increased glucose uptake and increased mitochondrial oxidation. This represents a unique cellular metabolic phenotype that may mechanistically explain obesity- associated T cell dysfunction. Interestingly, weight loss was unable to normalize adipose inflammation, reverse altered T cell metabolism, or improve the impaired immune response to influenza in obese mice. In contrast; systemic treatment of obese mice with metformin reversed CD4+ T cell metabolic dysfunction and improved survival following influenza infection. The clinical relevance and importance of this finding are supported by multiple observational and retrospective studies over the last few years showing that patients taking metformin have reduced disease severity and mortality to both influenza and COVID. Multiple lines of evidence point to a key role for metformin in regulating T cell immune responses. First, metformin has been found to alter the gut microbiome, which we know to influence both tissue-specific and systemic inflammation, and thereby influence T cells indirectly. Second, metformin has been shown to attenuate several inflammatory diseases by regulating the balance of regulatory and effector T cells. Third, we have generated preliminary data in our lab showing that metformin decreases oxidative metabolism, as well as the production of inflammatory cytokines in activated CD4+ and CD8+ T cells and differentiated Th1 and Th17 cells in vitro, indicating a direct effect of metformin on T cells. Therefore, the overall objective of this proposal is to elucidate the mechanisms by which metformin regulates T cell metabolism and function, directly and indirectly, using mouse models and human samples. To do so, we will perform the following aims: (1) Identify changes in gut microbiome and inflammatory cytokines in obese mice treated with metformin and determine if these changes drive T cell responses; (2) Determine the molecular mechanisms by which metformin directly affects T cell metabolism and function; and (3) Test if treatment with metformin can reverse obesity-associated dysfunction in human T cells. Successful completion of these aims will identify mechanisms by which metformin regulates T cell metabolism and function and reveal novel targets to improve treatment to viral infection in patients with obesity.

NIH Research Projects · FY 2026 · 2024-12

Abstract Heart, lung, blood, and sleep (HLBS) disorders affect millions of Americans and result in significant costs to the US healthcare system. These disorders also increase the risk of developing other detrimental health con- sequences, such as diabetes, depression, and metabolic disorder. Although a large amount of data has been generated, limited progress has been made in combating HLBS diseases. Proper analysis of these invaluable data is crucial for in-depth understanding of causal clinical and physiological bases regarding complex diseases, leading to more effective interventions and sustainable disease management and prevention strategies. However, reliable analysis of practical biomedical data can be highly challenging due to observational study designs, which often leads to complex confounding structures. Also, HLBS disease etiologies are intricate, with various clinical, metabolic, neurochemical, and immune-inflammatory factors entangling to impact disease phe- notypes. Additionally, patient population, disease, and drug response heterogeneity constitute another critical challenge. These complexities could result in biased, inconsistent or contradictory conclusions when conven- tional analytical tools are adopted to analyze practical biomedical data. To address these critical challenges, this proposal aims to adapt a set of robust data analytic and modeling tools based on novel machine learning methods. Specifically, this study will 1) propose novel machine learning methods for adjusting complex con- founding structures to reveal deep causal relationships between clinical features and health/disease outcomes; 2) build original deep learning hidden subgroup analysis frameworks to deal with the complicated heterogeneity for creating optimal health management regimens tailored to individual subject’s needs; and 3) establish a flexi- ble and scalable feature importance test framework to identify important biomarkers for improving disease/health outcome prediction performance. These methods will be applied to the analysis and modeling of data from the Hispanic Community Health Study/Study of Latinos, the most comprehensive study of Hispanic/Latino health and disease in the United States. Ultimately, this project will elucidate an array of clinical bases of HLBS disorders at the population and individual levels, leading to improved human health.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY The solid tumor microenvironment (TME) imprints a compromised metabolic state in which tumor infiltrating lymphocytes (TILs) are unable to maintain effective energy synthesis for antitumor function and survival. CD8+ T cells in the TME must catabolize lipids via mitochondrial fatty acid oxidation (FAO) to supply energy in nutrient stress, with T cells enriched in FAO being adept at cancer control. However, endogenous CD8+ TILs and unmodified cellular therapy products fail to sustain bioenergetics in tumors, and the direct molecular mechanism that underlies this failure is unknown. Discovery of a molecular target that enables TILs to utilize effective antitumor metabolism could be implemented immediately to improve clinical immunotherapies. Using RNA- sequencing, lipidomics, confocal imaging, and spectral flow cytometry, we identified that abnormal lipid accumulation was associated with CD8+ TIL metabolic failure across multiple solid tumor types. Acetyl-CoA carboxylase (ACC) is an enzymatic switch that drives lipid accumulation in nutrient-replete states. Under nutrient limitation, ACC is inhibited to enable lipid catabolism through mitochondrial FAO. Paradoxically, we observed that the TME imposes perpetual ACC activity in CD8+ TILs, enforcing lipid storage that directly opposes FAO. Moreover, elevated ACC1 gene expression in tumor samples from melanoma and sarcoma patients was associated with poor survival outcomes. Strikingly, we found that restricting ACC wholly rewired T cell lipid utilization and metabolism, producing a T cell pool enriched in mono and polyunsaturated fatty acids that could be rapidly metabolized for energy, supporting T cell control tumor control. This research program will use genetic mouse models, tumor tissue from cancer patients with advanced disease, pre-clinical CAR-T cell mouse models, and CAR-T products infused into cancer patients to formally test that ACC is a crucial enzymatic switch that profoundly limits antitumor immunity, restricting cancer control. Aim 1 of this proposal will use WT littermate and CD8creACC1f/f mice paired with tumor growth studies, lipidomics, metabolomics, and free fatty acid (FFA) analysis to test that the solid TME engages ACC1 to induce steatosis and loss of antitumor function in endogenous CD8+ TILs. We will validate the clinical relevance of our hypothesis using primary tissue from cancer patients and stored samples from metastatic melanoma patients treated with PD-1 inhibitors. Aim 2 of this proposal will use pre-clinical human and immune-competent CAR-T cell models paired with lipidomics, metabolomics, FFA analysis, and tumor growth studies to test that limiting ACC optimizes CAR-T cell performance in solid tumors. We will validate the clinical relevance of our hypothesis using CAR-T cell products previously infused into cancer patients. Results generated from this proposal will formally establish that the solid TME enforces ACC expression in CD8+ TILs, undermining bioenergetic plasticity by enforcing lipid storage. The data will fill a long-standing gap in knowledge surrounding T cell metabolism in tumors, providing a revolutionary molecular switch able to enhance endogenous and cell-based immunotherapeutic potency.

NIH Research Projects · FY 2026 · 2024-12

Project Summary This application is for Juliet S. King to pursue a Predoctoral Fellowship from the Ruth L. Kirschstein National Research Service Award (NRSA, F31). Ms. King is currently investigating tissue specific signaling modules in the oral epithelia during mammalian palatogenesis. This award will allow Ms. King to hone her cell and developmental biology skills and apply that expertise to her interests in craniofacial and oral biology. This proposal will advance our understanding of 1) periderm specific signaling programs and 2) the developmental function of heterotypic nectin interactions at the periderm-basal cell interface. Tissue specific and mechanistic insights into palatogenesis will provide clarity on cleft palate (CP) pathology and inform better genetic therapy strategies. With this opportunity, Ms. King will receive the training necessary for a career as an independent researcher with a focus on mentorship and education. Cleft palate (CP) is a common birth defect, affecting 1:1700 live births globally1. CP results from the failure to fuse the palatal shelves to separate the oral and nasal cavities2. Palate closure is a complex morphogenetic process that occurs during embryogenesis in both mice and humans. Like other morphogenetic processes, palate closure relies on cell-cell adhesions to generate proper tissue shape3. NECTIN1 and ECAD are two types of cell-cell contact receptors that when mutated have been causally associated with CP in humans4–7. Recent work from our lab has shown that loss of AFDN, the NECTIN1 downstream partner, in the oral epithelia drives CP. Importantly, the oral epithelium is made up of two compartments: the basal keratinocytes and periderm. Until recently, these two compartments have been experimentally treated as one, obscuring the tissue specific roles of either in palatogenesis. Thus, the goal of my project is to elucidate the requirement of periderm-specific signaling and the role of receptor signaling at the periderm-basal cell interface. I plan to test the hypothesized model that NECTIN1 in the basal layer interacts with NECTIN4 in the periderm to mesenchymal fusion and periderm-basal cell attachment in palate closure. To do this, I will 1) perform periderm-specific gene deletion studies and determine its effect on MES dissolution during palate fusion. In addition, I will 2) disrupt the NECTIN1-NECTIN4 heterotypic boundary between periderm and basal cells to define the role of heterotypic NECTIN interactions in palatogenesis. Our experiments take advantage of specialized cell biological and microscopy skills to address knowledge gaps in oral biology.