Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2026 · 2025-06

Abstract Proper hedonic processing is vital to perform motivated behaviors, but it can be disrupted by different psychiatric illnesses (e.g., substance use disorder and depression). Such disruption can lead to the development of negative affective states prevalent in suicide victims and during the craving/withdrawal phase of SUDs, contributing to relapse. While neural mechanisms mediating negative affect in depression and SUDs overlap, the neural changes produced by one makes the brain more susceptible to the other illness, increasing the risk of comorbidity. Considering that neuropsychiatric disorders are associated with dysfunction within large-scale brain networks, the overall goal of this project is to understand the global impact of different kinds of learned negative affect in such network activity. To do so, I will use taste reactivity (TR) to measure affective states in rats, in which aversive and appetitive orofacial responses to an intraorally infused solution are analyzed. To study the development of learned negative affect, I will combine TR with conditioned taste aversion (CTA), in which pairing a sweet tastant (e.g., saccharin) with a malaise-producing agent such as Lithium Chloride (LiCl), can shift the affective response of the rat from appetitive to aversive. Interestingly, while LiCl CTA is developed exclusively due to the aversive properties of the drug, such conditioned aversion can also be developed to agents that are not a priori aversive, such as drugs of abuse (e.g., cocaine). Importantly, while LiCl CTA allows us to model negative affect associated to unequivocal aversive experiences, drug CTA allow us to model complex aspects of addiction, such as natural reward devaluation and cue-induced withdrawal. Additionally, it has been shown that affect in general, and negative affect in particular, are differentially encoded across sex. To study the underlying differences between LiCl and cocaine CTA in male and female rats, I propose 2 Aims in the mentored phase and 2 aims in the independent phase of this application. Aim 1 (K99 phase) of this proposal will use rodent functional magnetic resonance (fMRI) and TR to investigate the effects of LiCl CTA on resting state (RS) activity in large-scale brain networks, including the default mode network (DMN), the lateral cortical network (LCN) and the salience network (SN) in male and female rats. Aim 2 (K99 phase) will use a highly novel multi-spectral, four-channel fiber photometry method and TR to examine how key nodes within the DMN and SN respond to the taste infusion after LiCl CTA development in male and female rats. Aim 3 (R00 phase) will use fMRI and TR to study the impact of cocaine CTA on brain network RS activity. Aim 4 (R00 phase) will use TR and fiber photometry in key nodes of the DMN and the SN to examine their response to the taste cue after cocaine CTA development. Collectively, these studies will provide critical insight into how brain networks and key nodes within them are altered after the development of two distinct forms of negative affect (LiCl and cocaine CTA).

NIH Research Projects · FY 2026 · 2025-06

Porcine Related Approach to Multidisciplinary Collaborative Translational Model Research (PROMOTER) ABSTRACT Large animal models of human disease are essential to the development, testing, and translation of therapeutics, but fully characterized large animal models of digestive diseases are relatively lacking. Within this broad digestive disease category, esophageal diseases are an attractive use case, being increasingly prevalent but paradoxically understudied. There are no approved esophageal-specific drugs or delivery mechanisms, and fundamental questions remain about disease pathogenesis, natural history, outcomes, and individualization of care. There is a critical need for a large animal model “toolbox” for discovery work enabling rapid translation of knowledge to practice for esophageal diseases. Because small animal models do not recapitulate human esophageal micro- or macro-structures, are minute in size, and require sacrifice to obtain tissue for examination, progress in many areas of research is hampered. In contrast, the pig is an ideal esophageal model. The pig esophagus is analogous in structure and function to the human esophagus, with similar epithelial and sub-epithelial compartments, and motility. In addition, as the pig and human esophagus are of similar size, the pig model is amenable to endoscopic assessment, enabling both visual inspection and recovery of tissue samples without sacrificing the animal. This facilitates critical long-term modeling to study disease pathogenesis, progression, and outcomes that are of direct clinical interest. We have begun to establish the viability of the porcine esophageal approach as a novel resource by exploring injury/repair, the role of stem cells, and drug delivery (with both novel mucosal-targeted formulations and drug-eluting devices), and have created a porcine disease model for eosinophilic esophagitis which demonstrates clear translational potential to human disease. With our established multidisciplinary team, we propose to use the RC2 mechanism to accelerate discovery and translation to the clinic in esophageal diseases and ultimately in other organs. Our application has been designed to meet all objectives of the High Impact Specialized Innovation Program, as well as multiple CTSA program goals, with the following specific aims: 1) To generate a comprehensive transcriptional, immunologic, and proteomic profile of the normal porcine esophagus; 2) To develop and generate comprehensive transcriptional, immunologic, and proteomic profiles of porcine models of inflammatory/injury-induced and fibrotic esophageal diseases; and 3) To establish an integrated and self- sustaining center that shares resources with the scientific community to promote translational multidisciplinary research, fundamental hypothesis testing, and therapeutic drug and device development in esophageal diseases, as an initial platform for extension to other human diseases. This highly innovative and impactful discovery-based and hypothesis-generating research will overcome roadblocks in translational science and enhance activities of the UNC/NC State CTSA UM1 Hub, successfully address a major impediment to translational science, and advance translation of drugs and devices to the clinic.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY/ABSTRACT Malaria in pregnancy (MiP) carries substantial - but largely preventable - risks for both the pregnant woman and unborn child. Despite rigorous evidence supporting the efficacy of current interventions, such as long-lasting insecticidal nets (LLIN) and intermittent preventive treatment of malaria in pregnancy (IPTp), MiP is estimated to occur in more than 40% of livebirths in endemic areas of sub-Saharan Africa. Therefore, additional interventions that would complement existing strategies are urgently needed One intriguing, but largely unexplored strategy for preventing MiP would be to reduce the risk of malaria transmission in the home environment, essentially creating a “safe zone” around the pregnant woman. This strategy has many potential advantages, including (i) leveraging the spatiotemporal clustering of malaria risk, (ii) bridging gaps in the existing package of interventions (i.e., imperfect LLIN use and missed IPTp doses), and (iii) the established safety and effectiveness of mass drug administration (MDA) programs. Therefore, the scientific objective of this Exploratory/Developmental Grant (R21) application is to demonstrate the feasibility, acceptability, and preliminary effectiveness of a focal mass drug administration (fMDA) program for household members of pregnant women to protect against MiP. Our hypothesis is that eliminating the parasite reservoir within the household will provide a complementary layer of protection against MiP especially when access to care is limited and visits may be delayed or missed. To achieve this, we will conduct a pilot randomized controlled trial to: Aim 1: Determine the feasibility and acceptability of a fMDA program with dihydroartemisinin- piperaquine (DP) as a novel component of the MiP prevention package. We will conduct an open-label, randomized pilot study at a primary health center in rural western Uganda. Women presenting to their first antenatal clinic visit will be randomized 1:1:1: to (i) monthly fMDA, (ii) one-time fMDA, or (iii) usual care. Using an established implementation framework, we will assess process measures such as the proportion of household members reached, adherence to the course of treatment, and frequency of adverse events. Aim 2: Estimate the efficacy of fMDA to create a “safe zone” in the immediate home environment and ultimately prevent MiP. Using the study design outlined in Aim 1, we will follow participating pregnant women and associated households through delivery, including longitudinal assessments of P. falciparum infection. As a pilot study, the trial is deliberately not powered for statistical tests of significance, but we will measure the incidence of (i) clinical malaria, defined as the presence of typical symptoms (e.g., fever, lethargy) and a positive malaria rapid diagnostic test (RDT), (ii) asymptomatic P. falciparum parasitemia and placental malaria by PCR throughout pregnancy and (iii) the incidence of adverse birth outcomes (e.g., stillbirth, low birth weight). In addition, we will measure the prevalence of asymptomatic parasitemia in household members using RDTs at three time points in order to estimate the effectiveness of fMDA at maintaining a parasite free zone.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY/ABSTRACT Perimenopausal women face an increased risk of developing major depressive disorder (MDD), with anhedonia as a central symptom. Anhedonia during perimenopause is linked to fluctuations in estradiol levels (“estradiol flux”), potentially influencing reward responsiveness through dopamine modulation in the mesolimbic pathway. No research has yet explored this relationship using simultaneous PET-MR, a tool uniquely suited for its molecular specificity. This proposal aims to bridge this knowledge gap by examining the neurobiological mechanisms connecting estradiol flux and anhedonia during perimenopause. Our objectives are 1) to examine the influence of estradiol flux on striatal dopamine binding using [11C]raclopride PET, 2) to study the relationship between estradiol flux and neural connectivity during reward processing via fMRI, 3) to assess the influence of estradiol flux on self-reported anhedonia symptoms, and 4) to explore the mediating roles of dopamine binding and brain connectivity between estradiol flux and anhedonia. This study complements an ongoing NIMH project (MH128238) by focusing on estradiol variability's effects on dopamine metabolism, brain reward response, and anhedonia in perimenopausal women.

NIH Research Projects · FY 2025 · 2025-06

The inflammation-resolving, analgesic, and anxiolytic properties of lipid derivatives of marine omega-3 (n-3) fatty acids offer a novel therapeutic approach for painful temporomandibular disorder (TMD). We used ultra- high performance liquid chromatography (UHPLC) tandem mass spectrometry methods to quantify fatty acids and their lipid derivatives in 605 adults. We reported that low levels of n-3 fatty acids were associated with greater odds of chronic TMD pain, hyperalgesia, psychological distress, sleep disturbance, and greater pain intensity. While promising, these observational study findings lack the rigor of a randomized controlled trial. Design: This phase 2b, proof-of-concept, triple-masked, randomized, placebo-controlled, parallel-group clinical trial will evaluate the safety and analgesic efficacy of a marine oil supplement in adults with chronic TMD myalgia and/or arthralgia. The intervention is a dietary supplement of active fractionated n-3 marine lipid concentrate standardized to 18-HEPE, 14-HDHA and 17-HDHA, which are precursors to specialized proresolving mediators (SPMs) that regulate multiple pain-related biochemical pathways. The primary endpoint is postbaseline change in pain intensity recorded on a 0-100 numeric rating scale. The sample size of 100 provides 80% power to detect a “moderately important” decrease of 34% in pain intensity in the intervention group compared to a “minimally important” decrease of 11% in the placebo group. UG3 planning phase: We will obtain regulatory approvals and develop study documents and data systems needed to implement the trail, optimize recruitment with CTSA resources, and refine UHPLC tandem mass spectrometry analysis for targeted profiling of 67 bioactive oxylipins and SPMs in erythrocytes and plasma. UH3 implementation phase: UH3 Aim 1 will quantify safety by comparing the rate of adverse events between study arms. UH3 Aim 2 will compute change in pain intensity in each treatment arm and estimate efficacy using two approaches in intention-to-treat analyses: (i) a linear mixed model will estimate treatment-group difference in mean post-baseline change in pain intensity; and (ii) a log binomial model will estimate treatment- group difference in the proportion of subjects whose mean pain intensity reduces by at least 30%. UH3 Aim 3 will evaluate reductions in hyperalgesia, psychosocial distress, sleep disturbance, and biochemical changes. We anticipate that the intervention will increase concentrations of proresolving, antinociceptive and anxiolytic oxylipins, and decrease omega-6 derivatives, and that these changes will correlate with changes in the clinical endpoints. UH4 Aim 4 will use causal mediation methods to determine if a therapeutic effect (or non-effect) is due to change (or lack of change) in nociception, psychological distress, sleep disturbance, or headache. Impact: Ten million U.S. adults have chronic TMD pain. This clinical trial may lead to a novel therapeutic approach for chronic TMD pain that resolves inflammation and reduces pain, sleep disturbance and psychological distress while overcoming the efficacy and tolerability limitations of current therapies.

NIH Research Projects · FY 2025 · 2025-06

PROJECT ABSTRACT Polysubstance-involved overdose rates have risen across the United States. Gabapentin, an anticonvulsive medication with FDA-approved uses for postherpetic neuralgia and partial seizures, is a co-substance of growing concern. Despite its limited on-label indications, gabapentin prescriptions have steadily increased, likely driven by off-label use. Reports of recreational abuse and misuse, particularly among individuals with opioid use disorder, are of particular concern, given that concurrent gabapentin and opioid misuse may result in death. Gabapentin has been increasingly detected in fatal overdose toxicology, especially in the Southeast United States. These harms have led to calls for increased monitoring or scheduling of gabapentin. In June 2018, West Virginia (WV) became the second of eight states to classify gabapentin as a schedule V controlled substance; however, the net effectiveness of the policy is unknown. Policy changes concerning controlled substances, which often result in altered prescribing patterns, have the potential to address concerns like over-prescribing and misuse but may also endanger users with dependence. Limiting access to prescribed medications may result in a shift to alternative substances obtained from nonmedical sources with unknown potency and potentially high levels of contaminants. This study will utilize a controlled interrupted time series design to assess the impact of the policy in WV, using North Carolina (NC) as a control state to account for secular changes given the lack of controlled substance designation in that state. Using WV’s Forensic Drug Database and NC’s State Unintentional Drug Overdose Reporting System, Aim 1 will assess whether gabapentin’s schedule V controlled substance classification in WV was associated with a change in 1A) the rate of gabapentin involved fatal overdoses, and 1B) the overall fatal overdose rate. Using Merative MarketScan Commercial Claims and Encounters Databases, Aim 2 will assess whether gabapentin’s schedule V classification in WV was associated with a change in prescription utilization among 2A) all privately-insured adults and 2B) adults with diabetic neuropathy for whom gabapentin is clinically indicated. Using Medicaid’s State Drug Utilization Data, Aim 3 will assess whether gabapentin’s schedule V classification in WV was associated with a change in outpatient gabapentin prescriptions paid for by Medicaid. Our longitudinal data analysis approach will allow us to identify both immediate effects and trend changes resulting from the scheduling policy’s implementation. As more states, as well as national policymakers, consider classifying gabapentin as a schedule V controlled substance, it is imperative to understand the impacts of WV’s implementation, both intended and potentially unintended. Results from this study will enhance the evidence base of gabapentin’s schedule V classification and inform future policy discussions.

NIH Research Projects · FY 2025 · 2025-06

Abstract This is an application for 2025-2026 bridge funding for the Intellectual and Developmental Disabilities Research Center (IDDRC) at the University of North Carolina (UNC). The UNC IDDRC is an interdisciplinary program with the overarching goal of supporting and promoting research relevant to understanding the pathogenesis and treatment of neurodevelopmental disorders. The UNC IDDRC is a critical component of the Carolina Institute for Developmental Disabilities (CIDD), which is the focal point for clinical services, research, and training relevant to intellectual and developmental disabilities (IDDs) on the UNC campus. The UNC IDDRC currently supports 55 investigators from 13 University departments, and includes 110 externally-funded research projects. The broad-based research program of this IDDRC is integrated around three themes: (1) Brain and Behavior Development; (2) Autism and Related Neurodevelopmental Disorders; and, (3) Early Detection/Intervention (including Clinical Trials), and the cross-cutting themes of Genetics/Genomics and Environmental Influences on IDDs. The UNC IDDRC is a highly collaborative program highlighted by the presence of several NIH-funded, interdisciplinary research centers/programs. This application seeks support for the same activities as the currently funded center: (1) an Administrative Core; (2) three research cores – the Clinical Translational Core, which includes the Research Participant Registry and Brain-Behavior Measurement Laboratory; the PreClinical Core, which includes the Preclinical Microscopy Core, Preclinical MRI Core, and the Preclinical Behavioral Core; the Data Science Core Core; and (3) a signature research project (a continuation of the project in the currently funded center) to study the pathophysiology of CSF and the meningeal lymphatic system in idiopathic autism, Fragile X syndrome, and Angelman syndrome by examining the mechanisms and clinical links to increased EA-CSF in etiologically and phenotypically distinct IDDs, and their mouse-model counterparts. This project has the potential to elucidate novel directions for targeted treatment development and to identify novel predictive biomarkers. The three proposed research cores provide cutting-edge, high-quality and cost effective support for this integrated, multidisciplinary, translational program of IDD-relevant research. Overall, this Center has had a major impact on IDD research and scientific training at the University of North Carolina, and renewal would allow us to continue our support of the truly exceptional program of IDD research conducted at UNC.

NIH Research Projects · FY 2026 · 2025-06

Prenatal alcohol exposure (PAE) alters gene expression and brain development and causes cognitive and behavioral impairment in humans and in animal models, and there is great interest in interventions that ameliorate these. One intervention showing efficacy is supplementation with the nutrient choline. Higher maternal choline status during pregnancy is positively associated with behavioral and cognitive improvements that persist throughout childhood, and both prenatal and postnatal choline treatment mitigate behavioral deficits in animal models of PAE and improve outcomes in people with Fetal Alcohol Spectrum Disorders. It also protects against a PAE-induced reduction in brain weight in mouse PAE models, but it is not known how choline improves brain development under PAE or if it affects crucial biological processes including cell proliferation and apoptosis. To gain insights into the potential mechanisms, we performed transcriptomics and recently reported that PAE alters the expression of multiple genes in the Hippo signaling pathway, which is crucial for brain development and growth due to its transcriptional regulation of cell proliferation. Complementary to this, in vitro studies in our lab show that PAE reduces proliferation and increases apoptosis of neural progenitor cells (NPCs), which can be regulated by the Hippo signaling pathway. Others have shown that this pathway is also influenced by maternal choline status, causing us to further investigate our transcriptomics analyses, to discover that choline supplementation during PAE reversed the impact of PAE on several Hippo components. In this proposal I hypothesize that the reductions in fetal brain size following PAE are due, in part, to increases in Hippo signaling that reduce the expansion of the neural progenitor cell population, and that choline improves brain development, in part, by normalizing those signals. These studies will generate novel insights into choline’s mechanism of action. I will test this using in vivo and in vitro techniques. I will use an established PAE mouse model where pregnant C57BL/6J mouse dams are exposed to alcohol and treated with choline, then fetal brains are collected at embryonic day (E)14.5. I will also manipulate Hippo signaling in a primary NPC culture model, followed by alcohol treatment in media having different choline concentrations. Experiments in Aim 1 use E14.5 fetal brains for anatomical analysis of cell proliferation and apoptosis and Hippo signaling components, then targeted qPCR and western blot to quantify the Hippo components that are dysregulated by PAE and mitigated by prenatal choline supplementation. Aim 2 uses transfected NPCs to induce gain- and loss-of-function of Hippo components to identify the role of Hippo pathway in alcohol’s effects on cell proliferation apoptosis and how choline remediates. This project fills a critical gap in understanding how prenatal choline supplementation mitigates the deficits in brain development caused by PAE and will help clarify the role of choline in normal brain development as well as the need for increased choline consumption during pregnancy to minimize the impact of stressors such as alcohol.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY/ABSTRACT Nearly 6 million people worldwide are infected with the parasite Trypanosoma cruzi, the agent of Chagas disease, and vertical transmission accounts for 22% of all new infections. Between 5-10% of T. cruzi-infected women will transmit the infection to their children, leading to a spectrum of disease in the infant: the majority of infected infants are asymptomatic but up to 40% have systemic disease involving the heart, brain, or liver. Congenital infection can also result in premature birth, or, in rare cases, neonatal death. Nearly a third of infants, even those with no symptoms, will develop chronic cardiac and gastrointestinal problems later in life. Unfortunately, up to 75% of affected infants do not receive timely diagnosis or treatment. Most T. cruzi infections occur in resource-limited rural areas of Latin America where advanced diagnostic tools are unavailable. Because treatment is safer and more effective during infancy than in older children and adults, early diagnosis is critical. Infected infants will continue to miss their opportunity for lifesaving treatment until we develop better algorithms to identify high-risk infants and accurately diagnose congenital Chagas disease in the perinatal period. Our long-term goal is to accelerate the elimination of congenital Chagas disease by developing tools to predict and diagnose T. cruzi infections in infants. In this project, we propose to (1) identify maternal clinical and epidemiological risk factors for vertical transmission; (2) investigate the parasite’s mechanisms of invading through the placenta using parasite genotyping and identifying gene expression signatures associated with transmission; and (3) optimize and validate a new test for congenital T. cruzi infection using recombinase polymerase amplification (RPA), a DNA detection method that can be performed without advanced laboratory equipment or expertise and is adaptable to point-of-care platforms. To accomplish these goals, we propose a cohort study of pregnant women and their infants in Santa Cruz, Bolivia, a highly endemic area for T. cruzi. We hypothesize that by combining traditional epidemiological analyses with cutting- edge genomic techniques, we can better predict which infants born to women infected with T. cruzi are at highest risk of congenital infection. We also hypothesize that a new RPA assay will allow us to diagnose these infants more frequently and earlier than current standard diagnostics, which perform poorly to detect congenital infection in neonates. In Aim 1, we will identify risk factors for vertical transmission in a cohort of T. cruzi- infected women to better identify high-risk infants. In Aim 2, we will characterize parasite genetic factors associated with vertical transmission of T. cruzi using genotyping and transcriptomics. Finally, in Aim 3, we will optimize our RPA assay and test it in the clinical setting to evaluate its ability to diagnose neonates with congenital Chagas disease. Our innovative approach will integrate field epidemiology, cutting-edge sequencing techniques, and new diagnostics to identify infected infants in the first weeks of life. These advances have the potential to lead to a paradigm shift to accelerate cure of Chagas disease’s youngest victims.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY Dr. Joseph Burclaff, PhD is mentored by Dr. Scott Magness, PhD at the University of North Carolina at Chapel Hill. UNC receives the 6th highest federal research funding of any American university and is home to the NIDDK-sponsored Center for Gastrointestinal Biology and Disease with a vibrant research community. The CGIBD and Dr. Burclaff’s home department of Biomedical Engineering provide supportive environments for young investigators. Dr. Burclaff proposes an innovative cross-disciplinary research plan bridging human in vitro genetic engineering, in vivo mouse work, functional metabolic studies, inflammatory signals, and transcription factor analysis, all within the frameworks of homeostasis and inflammatory bowel disease (IBD). Dynamic regulation of mitochondrial metabolism is essential for healthy intestinal stem cell (ISC) activity, with mitochondrial dysfunction known to drive inflammatory signaling and aberrant cell fates able to initiate experimental colitis. IBD is marked by decreased mitochondria, increased proinflammatory signaling, and altered ISC cycling, and mitochondrial and pro-inflammatory signaling are even seen in non-inflamed tissue from IBD patients, implicating mitochondrial dysfunction as an early event in IBD development or even predisposing the intestinal epithelium for inflammation. Few transcription factors (TFs) are known to regulate mitochondrial dynamics in ISCs. My work with primary human ISCs genetically engineered to knockout or induce overexpression of the TF SOX9 show that SOX9 strongly represses mitochondrial activity, slows cell cycle, and increases proinflammatory gene expression in ISCs. The specific effects of SOX9 on mitochondria and whether SOX9 regulates inflammatory signaling and ISC activity directly or by acting through mitochondria are unknown. This proposal will determine how SOX9 regulates mitochondria and proinflammatory signaling in human ISCs and in mice. As SOX9 often acts through other TFs, and SOX9 appears to regulate mitochondria without affecting previously implicated mitochondrial-related TFs in these ISCs, candidate TFs identified using this primary human intestinal platform will be tested for their ability to regulate mitochondria and ISC activity independently or as a TF network. Cutting-edge cellular engineering strategies will further test direct downstream targets of these TFs to determine mechanisms by which this novel network regulates mitochondria in the human intestinal epithelium. To establish an independent research area in this field, Dr. Burclaff requires mentored time to become fluent in metabolism and metabolomics, TF interactions with each other and downstream genomic targets, and IBD development. To achieve this, Dr. Burclaff will be co-mentored by Dr. Costas Lyssiotis at the University of Michigan, a field expert in metabolism and metabolomic analysis in gastrointestinal tissues and disease states. Dr. Burclaff also assembled a committee of field experts in metabolism, chromatin and TF analysis, and IBD to help guide him through this project and in his transition to establish an independent research program to train the next generation of researchers and to study regulatory mechanisms of ISCs that affect human health.

NIH Research Projects · FY 2026 · 2025-05

Bacterial pneumonias remain important clinical problems with major morbidity and mortality. S. pneumoniae is the leading cause of community-acquired pneumonias and an important cause of ARDS and post-viral bacterial pneumonias. The lung's immune response is critical to clearance of S. pneumoniae, repair of the lung tissue, and return to homeostasis. Our studies have addressed many aspects of neutrophil kinetics and function and document the numerous ways in which neutrophils vary and can be categorized, based on cytokine production, surface markers, transcriptomes, age, or many other criteria, each of which describes the range of functions that neutrophils can perform. This proposal focuses on the mechanisms underlying the process through which neutrophils function to identify and repair the infection and injury and/or to further injure the lungs. Our overall working hypothesis is that during the immune response, neutrophils undergo changes in their transcriptomes that regulate their function and that the unique structure of the lungs creates microenvironments within the parenchyma and the alveoli/small airways that regulate the phenotype of neutrophils. We further postulate that spatial and temporal features of the innate immune response contribute to this phenotype. Aim 1 will determine the changes that neutrophils undergo as they are recruited and carry out their functions during S. pneumoniae pneumonia and the biological importance of these changes. We will determine the changes in the transcriptome of neutrophils in the systemic blood, pulmonary capillary blood, lung parenchyma and alveoli/airways at 6h, 18h, 2d and 4d after inoculation, identifying both spatial and temporal changes in neutrophil transcriptomes using scRNAseq/CITE-seq. We postulate that heterogeneity in endothelial and alveolar Type 1 and 2 cell transcriptomes will describe niches of migration. Protein expression of identified neutrophil proteins will be assessed using mass cytometry. The mechanisms underlying the phenotypes of neutrophils expressing IFNγ, IL-1α and/or TNF and of SiglecF+ neutrophils will be studied. Aim 2 will determine the changes in the lung microenvironment during the development and resolution of S. pneumoniae-induced inflammation and injury. The spatial transcriptome of alveoli and bronchioles/bronchi will be determined 6h, 18h, 2d and 4d after inoculation of S. pneumoniae. The effect of signaling induced by Type I and II IFNs, IL-1α/β and TNF on neutrophil transcriptomes and function will be identified. The hypothesis that the oxidative microenvironment experienced by myeloid cells contributes to transcriptomic heterogeneity and function will be tested. These studies will determine the changes in neutrophils as they function during pneumonia, will compare neutrophils defined by particular criteria, and will shed light on the changes and contributions of the lung microenvironment as inflammation develops and resolves. They will reveal novel context-dependent relationships and interactomes, providing insight into neutrophil function in the innate immune response and potential therapeutic opportunities for bacterial pneumonia.

NIH Research Projects · FY 2026 · 2025-05

Research has demonstrated that exposure to ambient extreme heat is associated with increased morbidity and mortality. Extended periods of exposure to high temperatures are associated with causing conditions such as heat exhaustion, heat cramps, and heat stroke as well as exacerbating existing conditions including those related to the respiratory, cerebral, and cardiovascular systems. There is little research, however, on the relationship between heat exposure and cardiovascular health (CVH), especially among children. Further, research has not investigated how the vulnerability of neighborhood built and social environments to ambient heat may attenuate such a relationship. This study will address the gaps present in the literature by examining how extreme heat exposure is associated with blood pressure, a key indicator of CVH, during early childhood. It will also explore how neighborhood heat vulnerability acts as both an exposure impacting CVH and a modifier on the pathway between heat exposure and CVH. To do so, this study will leverage data from a regional birth cohort with follow-up through early childhood which will be linked to validated, national data for ambient, built, and social environmental exposures. In Aim 1, I will describe trends of how gestational extreme heat event (EHE) exposure and neighborhood vulnerability are distributed across the study area and by sample sociodemographic characteristics. In Aim 2, I will investigate associations between gestational EHE and blood pressure outcomes throughout early childhood. Finally, in Aim 3, I will investigate the role of gestational heat vulnerability with EHE and blood pressure. This study will advance our understanding of the mechanisms by which environment can affect CVH. This will inform future interventions and policy to reduce CVH morbidity from a growing public health threat during a sensitive period of life when prevention may be most effective.

NIH Research Projects · FY 2026 · 2025-05

ABSTRACT The molecular and cellular processes that control axonal degeneration in response to genetic mutations are poorly understood, hindering the advancement of regenerative strategies to improve patient outcomes. In the pediatric neurodegenerative disease giant axonal neuropathy (GAN), axons degenerate due to loss-of-function mutations in the gene KLHL16. Current understanding is that gigaxonin, the product of KLHL16, facilitates the proteasomal turnover of cytoskeletal intermediate filament (IF) proteins through its major function as an adaptor between an E3 ubiquitin ligase and the IF substrate. This mechanism explains the progressive accumulation of IF proteins in many cell types of GAN patients, but the reasons behind the selective and profound susceptibility of neurons to KLHL16 mutations are unknown. Seeking to fill this knowledge gap, we made the novel discovery that retinoic acid (RA) signaling is dramatically impaired in GAN patient neurons. RA is a critical molecule involved in nervous system development and regeneration, but its high regenerative potential has not yet been realized in humans. Our team has a unique opportunity to advance RA biology in the context of GAN using a focused approach centered on two disease targets we identified: the cellular retinoid acid binding protein-1 (CRABP1) and the RA receptor β (RARβ). Using a robust and clinically relevant experimental pipeline of GAN patient iPSC-motor neurons and a new humanized GAN mouse models that we developed, this project will examine the novel concept that targeted modulation of RA signaling will enhance axonal regeneration and functional improvement in GAN.

NIH Research Projects · FY 2026 · 2025-05

Human microbiomes hold promise as therapeutic targets, yet they remain under-exploited because of limited tools available to dissect the mechanisms underlying the complex microbiome-disease relationships, as well as tools that can precisely modify specific commensal bacteria. To fully realize the promise of microbiome- targeted therapies, methods that enable safe, efficient and highly-targeted transduction of specific bacterial species within complex microbial communities in situ are sorely needed. Due to their target cell specificity, bacteriophages (phages) carrying transgenes represent ideal vectors for imparting precise genetic control over bacterial consortia, similar to viral gene vectors currently used for human gene therapy. However, current phage vectors, which can be categorized as either replicative or non-replicative, suffer from a number of shortcomings that preclude their use in vivo. Replicative vectors, which packed both transgenic DNA and genomic DNA essential DNA for replication, are limited by a small transgene packing capacity, toxicity, and major risks for horizontal gene transfer. In contrast, non-replicative phage vectors (NRPV) are packed exclusively with transgenic DNA, which greatly increases transgene packing capacity while eliminating the risk of horizontal gene transfer. Unfortunately, NRPVs suffer from very poor delivery efficiency (<0.2%) that have precluded their use to date. To overcome these limitations of NRPVs, we have combined cutting edge synthetic biology with phage engineering to develop a new category of NRPVs that can achieve efficient self- recircularization upon the injection of its linear cargo DNA into bacteria host. As a result, the DNA delivered by our self-circularizing NRPV (scNRPV) is (a) not at risk of exonucleases degradation, and (b) can replicate with the target bacterial host as they divide, leading to sustained retention and expression of the transgene. The end result is orders of magnitude improvement in the delivery efficiency, capable of transducing ~80% of E. coli at just 1:1 ratio of scNRPV:bacteria in vitro, and achieving transduction efficiencies in vivo (>108 CFU/g) that rivals replicative phage vectors. As a novel delivery platform, we seek to perform key proof-of-concept data in vitro and in vivo. In Aim 1, we will produce and characterize various scNRPVs, including both T7- and P1- based scNRPVs. In Aim 2, we will benchmark the delivery efficiencies of (i) T7 scNRPVs in vivo, comparing them against (ii) replicative phages, (iii) conventional NRPV, and (iv) live engineered bacteria for delivery of mScarlette/luciferase as well as short-chain fatty acids, an important molecule whose absence is directly correlated to an increase in intestinal permeability and inflammation in inflammatory bowel disease. The proposed work will provide key proof-of-concept motivating further explorations using scNRPVs for in situ microbiome engineering. We expect our work will open the door for investigators to utilize these engineered phages as tools to perform targeted genetic manipulations with large transgene cassettes of specific species in complex microbiomes both in vitro and in vivo for the first time, without the risk of horizontal gene transfer.

NIH Research Projects · FY 2024 · 2025-05

PROJECT SUMMARY / ABSTRACT Coordinated specialty care (CSC) provided in the early course of psychosis can reduce the long-term chronicity and dysfunction associated with schizophrenia-spectrum disorders. However, few young adults with early psychosis (EP) are engaged with these programs. Young adults with EP are particularly susceptible to stigma and negative beliefs about psychiatric treatment, and thus are often apprehensive to present to in- person care. Despite their reluctance to seek services, young adults with EP often engage with mental health resources through online and mobile technologies. Several digital health interventions have been developed as adjuncts to in-person services for this population and have demonstrated feasibility, acceptability and preliminary efficacy. However, there are few available digital health tools designed for young adults with EP who are not connected to in-person care. In the absence of such tools, individuals often turn to social media or forums that often reify stigmatizing beliefs or discourage help seeking. Mobile health interventions appear particularly well suited to this population given the fact that mobile devices are the primary media source for most young adults and can provide individualized, real-time, real-place support to introduce users to psychosocial interventions involved in CSC. It remains unclear at present, however, whether and how remotely delivered mHealth can be leveraged to increase engagement in treatment for young adults with EP. The proposed research project proposes to develop and test an mHealth intervention designed to impact help-seeking beliefs through psychoeducation, stigma-reducing content, stress management exercises, and locally relevant treatment information delivered through audio, video, and automated motivational text interactions, and personalized through ecological momentary assessment (EMA). This project will take a user- centered design approach with individuals who are both engaged and unengaged in treatment, including (1) a contextual inquiry to understand barriers, interests and preferences related to mHealth and in-person treatment, (2) development, refinement and usability testing, and (3) a pilot randomized controlled trial assessing feasibility, acceptability and preliminary efficacy compared to an active control mHealth intervention providing only stress management. This research project will provide a central piece of the candidate's overall training plan. This K23 career development award provides training to support an independent research program developing interventions that reduce barriers to care for young adults with EP. This includes three training areas, including (1) mobile health targeting increasing mental health care engagement, (2) user-centered design, and (3) advanced longitudinal models and quantitative methods for digital health trials. These goals will be met through expert mentorship, didactic training, coursework, and the successful completion of the research project.

NIH Research Projects · FY 2026 · 2025-05

Oral squamous cell carcinoma (OSCC) is a significant public health problem, with approximately 400,000 new cases and 200,000 deaths per year worldwide. First-line treatments for OSCC typically include surgery and radiation, with chemotherapy added to enhance the efficacy of radiation (chemoradiation), and to prevent cancer recurrence. Radiation and oral cancer chemotherapeutics eliminate cancer cells largely by disrupting DNA replication to induce replication stress and DNA damage, suggesting the therapeutic benefit of conjunctive targeting of the DNA replication machinery. However, replication functions are essential for normal cell proliferation, leading to unavoidable toxicity that limits the true clinical potential. In this project, we explore a novel idea that the stress-specific function of an essential DNA replication factor can be targeted to effectively and selectively confer toxicity in OSCC. The heterotrimeric replication protein A (RPA) complex binds and stabilizes single strand DNA (ssDNA) via multiple oligonucleotide binding (OB) motifs. RPA-ssDNA not only mediates DNA replication, but also orchestrates replication fork stabilization, DNA repair and activation of the replication checkpoint. Interestingly, a unique OB motif at the N-terminus of the RPA70 subunit (RPA70N) mediates RPA’s binding to, not ssDNA, but multiple proteins involved in the replication stress responses. We hypothesize that this RPA motif is targetable to enhance OSCC treatment responses. RPA upregulation in OSCC has been associated with cancer resistance, and replication stress is a well-established cancer hallmark, supporting the potential selectivity of the treatment. We will develop new transgenic mouse models to study the impact of RPA upregulation and RPA70N mutations on OSCC progression and treatment responses. Combining with these mouse models, we will carry out proof-of-principle, pre-clinical evaluation of a novel small molecule inhibitor of the RPA70N OB motif in OSCC treatment. Our multi-PI team are poised to explore the proposed therapeutic idea, with complementary expertise in RPA, transgenic mouse models, and OSCC.

NIH Research Projects · FY 2025 · 2025-05

PROJECT SUMMARY Significance. Sleep problems, including short sleep duration and sleep disorders, elevate risk of a numerous chronic diseases, particularly cardiometabolic diseases (CMDs), and contribute to racial and ethnic health disparities. One possible mechanism linking sleep problems to CMDs is chronic, low-grade inflammation. However, existing evidence on sleep, chronic low-grade inflammation, and CMDs has primarily focused on non- specific inflammatory biomarkers that are less likely to be causal. Because upstream regulation of inflammation is mainly dictated by polyunsaturated fatty acids and their oxylipin derivatives (“oxylipins”), these molecules afford novel insights into the role of inflammation in sleep and CMDs. Oxylipins are also amenable to treatment. However, very few studies have examined the relationship between sleep problems, oxylipins, and CMDs. The handful of studies that are available are small or did not measure a comprehensive set of oxylipins. While research on oxylipins and CMDs is more abundant, these studies have lacked diversity, did not prioritize which of the highly correlated oxylipins are most likely causal, and are limited by confounding and reverse causation. Specific aims. The proposed research aims to triangulate causal and mediating effects of ~130 oxylipins with sleep problems, including insomnia, sleep apnea, and short sleep duration, and CMDs to identify shared and distinct inflammatory pathways in the well-characterized Hispanic Community Health Study/Study of Latinos (n=16,415). Aim 1 will identify oxylipins associated with insomnia symptoms, a common (prevalence=10-30%) threat to sleep health using tailored statistical methods that accommodate highly correlated data and protect against reverse causation and residual confounding. Aim 2 will extend aim 1 to include sleep apnea and short sleep duration, enabling the applicant to examine the consistency of oxylipins across sleep problems. Finally, integration of CMDs (hypertension, type 2 diabetes, and obesity), again using models that accommodate highly correlated data and protect against residual confounding and reverse causation, will provide some of the first insights into inflammatory pathways linking sleep problems and CMDs. Fellowship information. The applicant, Ms. Sarah Koenigsberg, is an epidemiology PhD student at the University of North Carolina at Chapel Hill with a background in biochemistry, cardiometabolic disease, and population-level research. The proposed research and training plan would occur under the guidance of Ms. Koenigsberg's mentorship team, which consists of experts in molecular, cardiometabolic, and sleep epidemiology, biostatistics, and large-scale computation. Under this award, Ms. Koenigsberg would receive training on sleep epidemiology and evidence triangulation methods to complement her existing skill set, as well as engage in professional development activities that would equip her to achieve her long-term goal of becoming an independent investigator at the intersection of cardiometabolic, molecular, and sleep epidemiology.

NIH Research Projects · FY 2026 · 2025-05

Project Summary Opioid use disorders have an enormous societal impact in the United States and opioid overdose deaths are now the leading cause of accidental death in the United States. The impacts of opioid use disorder have worsened over the past decade due to prevalence of high potency synthetic opioids, like fentanyl, that pose unique problems for clinical treatment. Opioid withdrawal is a key contributor to opioid use disorder because withdrawal acts as a negative reinforcer to drive continued opioid consumption. Opioid withdrawal consists of both somatic and negative affective behaviors, and long-term opioid use is associated with the development of anxiety and stress. Therefore, it is critical to understand how opioids alter circuits and neurotransmitters that regulate addiction-like behaviors and negative affect. The basolateral amygdala (BLA) is a key component of the stress response and encodes aversive memories. The BLA receives noradrenergic innervation from A2-NE neurons located in the nucleus of the solitary tract, and this cell population is activated by opioid withdrawal. Notably, chronic stress disrupts noradrenergic plasticity in the BLA, and altered NE signaling in the BLA contributes to negative affect and disruptions in conditioned fear extinction. Despite these links, this pathway has been relatively underexplored in the context of opioid use disorder. My preliminary data demonstrate that A2-NE neurons are hyperactive and hyperexcitable following morphine withdrawal. I developed a mouse model of chronic oral fentanyl consumption, where mice consume escalating concentrations of fentanyl for 5 weeks in a drinking in the dark (DiD) model. I observed sex differences in withdrawal behaviors following naloxone- precipitated withdrawal. Following fentanyl consumption, BLA principal neurons had reduced excitatory/inhibitory balance, driven primarily by reduced inhibitory inputs, and increased excitability. Finally, I found that conditioned fear extinction is reduced following fentanyl consumption, while fear learning itself is intact. These data inform the central hypothesis of my proposal that will be investigated in 3 aims: 1. Patterns of fentanyl consumption and withdrawal induce changes in brainstem A2-NE neurons to drive increased excitability and tonic activity (Aim 1). 2. Increased excitability of A2-NE neurons drives increased NE release in the BLA, which disrupts BLA NE signaling (Aim 2). Finally, we hypothesize that increased NE neurotransmission and altered BLA NE signaling drives extinction learning deficits following fentanyl consumption (Aim 3).

NIH Research Projects · FY 2026 · 2025-05

Abstract: Sickle cell disease (SCD) is an inherited chronic hemolytic anemia characterized by vaso-occlusive episodes and end organ damage. Up to 80% of patients with SCD will develop chronic kidney disease (CKD), with 30% mortality ascribed to kidney failure. This devastating health outcome exists because there are numerous gaps in our understanding of complex mechanisms underlying SCD nephropathy. In patients and murine SCD, an increased renal iron deposition occurs primarily in proximal tubules (PT), and correlates with albuminuria, an early marker of CKD. Kidney-specific etiology of iron accumulation is purely a consequence of intravascular hemolysis, not chronic blood transfusion. Current interventional strategies are limited to the chelation of transfusion-mediated iron overload only. Classic iron chelators are nephrotoxic, and there are no available strategies targeting kidney iron overload, thus there is an unmet need for novel iron overload-modifying reno-protective strategies. However, molecular mechanisms of renal-specific iron handling remain to be elucidated. My preliminary studies indicate that ET-1 promotes PT iron accumulation. I observed a strong positive correlation between plasma ET-1 levels and renal iron accumulation in both mice and patients with SCD. Furthermore, selective ETA receptor antagonist reduces iron deposition in the kidney, at the same time increasing urinary iron excretion and serum iron levels in SCD mice. Third, my in vitro data show that ET-1 directly increases the expression of transferrin-1 receptor (iron uptake transporter) and decreases the expression of ferroportin-1 (FPN-1, iron exporter) in mouse PT cells. Together, these observations point to ET-1/ETA signaling as a novel regulator of renal iron handling in SCD. Based on these novel preliminary findings, we hypothesize that ET- 1/ETA activation promotes renal iron overload via direct increase in heme/iron uptake and/or inhibition of iron export in SCD. Using primary mouse and human PT cells, we will determine whether ET-1/ETA receptor activation increases heme/iron uptake via direct and/or HIF-1a-mediated activation of NF-kB signaling pathway. Using novel humanized SCD mice lacking PT ETA expression, we will determine the effect of ETA receptors deletion on renal iron burden and kidney health. Second, we will define specific ET-1/ETA-dependent control of FPN-1 and iron export in vitro, and in vivo mechanisms contributing to ETA-mediated renal iron recycling using acute hemolysis model. Lastly, we will determine the effect of ETA receptor antagonism on iron recycling and/or urinary excretion in SCD patients. We aim to unveil these molecular mechanisms with the following aims: Specific Aim 1: Determine the mechanism(s) by which ET-1/ETA signaling promotes iron uptake by PT and its subsequent contribution to SCD nephropathy. Specific Aim 2: Determine the mechanism(s) by which ET-1/ETA signaling reduces PT iron export, subsequently leading to iron accumulation in SCD. Specific Aim 3: Assess the relationship between renal iron handling and ETA antagonism in SCD patients.

NIH Research Projects · FY 2026 · 2025-05

Project Summary Trachoma is the leading infectious cause of blindness globally, and it disproportionally affects the world’s most marginalized populations. Trachomatous trichiasis (TT) is a condition resulting from years of repeated ocular chlamydia trachomatous infections that cause the eyelid to turn in and the eyelashes to rub the eye, eventually leading to blindness. While surgery is available to correct trichiasis, more than 20% of patients experience post-operative trichiasis (PTT) needing further surgical management. Yet, current management procedures for post-operative trichiasis are suboptimal. A recently completed clinical trial, BESRAT, compared an existing surgery procedure (posterior lamellar tarsal rotation) against a newly developed procedure, Bevel-Rotate Advancement Procedure (B-RAP). The trial used the best surgeons available. The trial suggested that B-RAP performed better than the existing procedure, particularly for patients with severe PTT or an eyelid contour abnormality at the time of repeat surgery. The next step is to determine whether a larger group of TT surgeons can perform B-RAP well and to compare it against the other standard TT surgery procedure, bilamellar tarsal rotation (BLTR). The primary objective of this randomized clinical trial is to determine whether repeat trichiasis surgery performed with B-RAP improves surgical success compared to BLTR among a group of 8-10 district TT surgeons in Tanzania. The study aims to enroll 1,000 individuals with PTT. The primary outcome is repeat PTT within one year after surgery. Additionally, the study will assess eyelid contour abnormalities and how they change over a two-year period as well as patient reported outcomes. If this project is successful in improving surgical outcomes, it could change the approach to treating PTT globally. Individuals with trichiasis have a significantly reduced quality of life; correcting their trichiasis long-term has the potential to improve their quality of life and their family members’ quality of life considerably.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY/ ABSTRACT Significance: Compared to peer countries, pregnant and laboring women in the US are more likely to experience unnecessary interventions during childbirth, making childbirth more expensive while putting patients at an increased risk of adverse pregnancy complications and mortality. A growing body of work estimates that between 16-20% of women in the US experienced a form of mistreatment during pregnancy care, and mistreatment may be associated with unnecessary clinical interventions during childbirth, birth trauma, and mortality. A person-centered approach may be an antidote to disrespectful care, over-intervention, and poor health outcomes during the perinatal period. TeamBirth is a person-centered care intervention during childbirth that puts the pregnant woman at the center of a team-based decision-making process. TeamBirth is currently being implemented across 55 hospitals in Washington state. While pilot studies have shown that TeamBirth is feasible and acceptable to implement among clinicians, no studies have tested the association of TeamBirth on clinical outcomes or pregnant women’s perceptions of care. Specific Aims: This study leverages an ongoing intervention across 55 hospitals in Washington state to estimate the impact of TeamBirth on (1) pregnant women’s perceived autonomy in decision-making during childbirth and (2) Cesarean delivery among low-risk pregnant women before and after implementation of TeamBirth. Approach: The proposed analysis will utilize data from two data sources: (1) patient surveys administered during in-patient postpartum care and (2) chart abstracted data from the Obstetrical Care Outcomes Assessment Program (OB COAP) and the Washington State Hospital Association Maternal Data Center (WSHA-MDC). Using a pre-post design and multiple linear regression, aim 1 will assess the change in pregnant women’s perceived autonomy in decision-making before and after implementation of TeamBirth. Aim 2 will use a retrospective birth cohort and log-risk regression to estimate the risk ratio and 95% confidence interval of Cesarean section given exposure to TeamBirth among low-risk patients. Fellowship Information: The applicant is a PhD student in Maternal and Child Health and has completed a minor in Epidemiology at The University of North Carolina in Chapel Hill. Ms. Spigel’s application is supported by a multidisciplinary team of mentors with expertise in perinatal nursing, person-centered care, perinatal epidemiology, applied microeconomics, evaluation of maternal health interventions, and implementation of TeamBirth. Through coursework in maternal health, epidemiology, quasi-experimental methods, coupled with support from an exemplary team of mentors, the proposed training plan will assist Ms. Spigel in successfully completing her dissertation work, and achieving her long-term goal of becoming a successful independent researcher with expertise in the quality and experience of care during the perinatal period.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY: The NRF2 transcription factor is mutated and activated in more than 30% of lung and upper aerodigestive squamous cell carcinomas. NRF2 drives a gene expression program that mitigates oxidative and electrophilic stress, reprograms and enables cancer cell metabolism, and suppresses lymphocyte infiltration. In lung squamous cell carcinoma (LUSC) patients, NRF2 activity portends a poor prognosis and resistance to radiation therapy, chemotherapy, and immune checkpoint inhibitors (ICI). Though frequently active in human cancers, mouse models suggest that NRF2 mutational activation is not sufficient for oncogenesis—the temporal and contextual underpinnings of how NRF2 contributes to cancer progression remain ill-defined. This research proposal focuses on a major research objective of our laboratories: to understand the impact of NRF2 activation in LUSC initiation and progression. To meet this goal, we have developed a novel genetically- engineered mouse model (GEMM) to test the impact of an activated NRF2E79Q on LUSC incidence, and on tumor histology, gene expression and metabolism. Because NRF2 activity correlates with resistance to ICI, our proposed studies will also assess how NRF2 alters the tumor microenvironment (TIME) in LUSC. Thus, we meet a research objective of PAR-22-216, “Development and early validation of in vivo or in vitro models for the assessment of immunoprevention or immunotherapy efficacy”. Based upon our previous studies along with other published reports, we hypothesize that NRF2 activation will increase LUSC incidence and progression, while decreasing mouse survival and altering the TIME landscape. To test this hypothesis, we propose two Specific Aims. In the first specific aim, we will define and understand NRF2’s oncogenic functions as a ‘second hit’ in LUSC initiation and progression in our novel Rosa26LSL-Sox2-IRES-GFP;Nkx2-1fl/fl, LSL-Nrf2E79Q (SNN) GEMM. In the second specific aim, we will assess the impact of NRF2 activation on TIME of LUSC developed in our GEMM. We will characterize the incidence and pathology of LUSC arising in our GEMM along with overall survival. We will apply state-of-the-art proteomics and mass spectrometry imaging to molecularly characterize NRF2-wildtype and NRF2-mutant tumors. We will also comprehensively define changes in the composition of pro- and anti-tumorigenic immune cell types and cell-intrinsic immunomodulatory molecules of the TIME using high dimensional spectral flow cytometry, OIS-PRM proteomics, IHC, and transcriptomics. We will use the data from this grant to support a subsequent R01 application identifying underlying mechanisms and therapeutic targets for NRF2-active cancers and determine the efficacy of pharmacological agents to suppress NRF2 as well as their abilities to sensitize LUSC to ICI therapy.

NIH Research Projects · FY 2026 · 2025-04

ABSTRACT The vaping landscape has significantly evolved since they were introduced, continuously gaining popularity among adolescents and young adults. Originally marketed as a smoking alternative or cessation aid, vaping devices now attract youth with various flavorings and addictive substances, including different forms of nicotine. Despite regulatory efforts, youth vaping remains a significant issue, especially with the rise of disposable devices like cool mint- and fruit-flavored vapes. Concerns about the safety of these devices compared to traditional cigarettes persist, as harmful compounds have been identified in vape emissions. The emergence of alternative nicotine formulations, such as 6-methyl nicotine, further complicates the situation. Research indicates that 6- methyl nicotine has similar physiological effects to nicotine but raises more significant safety concerns, necessitating further research and regulatory attention. Specifically, initial studies indicate that 6-methyl nicotine exhibits higher toxicity and greater activation of nicotinic receptors than nicotine, presenting a public health challenge due to gaps in understanding its long-term effects and potentially higher addictive attributes. To address these risks, this project hypothesizes that vaping-induced aerosolization of 6-methyl nicotine vaping products modifies the chemical composition of these e-liquids and generates toxic aerosol mixtures. The project aims to evaluate the chemical composition of 6-methyl nicotine and nicotinamide vapes before and after aerosolization and investigate their toxicity in human bronchial epithelial cells (hBECs), an initial target for inhaled e-cigarette aerosols. By integrating chemical characterization with in vitro exposures in our newly developed Vaping Product Exposure System (VaPES), these goals will be accomplished through two specific aims. Specific Aim 1 will analyze the chemical composition and metabolism of 6-methyl nicotine and nicotinamide vaping devices before and after vaping using targeted and nontargeted mass spectrometry. This will identify active ingredients, contaminants, and novel oxidation products formed during aerosolization. Specific Aim 2 will assess the toxicity of 6-methyl nicotine and nicotinamide compared to nicotine e-cigarettes in hBECs. This will involve evaluating cytotoxicity and gene expression, untargeted metabolomic and proteomic profiles in hBECs, and determination of chemical biomarkers of exposure. Computational modeling tools will be used to integrate chemical/transcriptomic/metabolic signatures associated with dysregulated cellular pathways in hBECs. The project's overarching goals are to provide comprehensive chemical characterization and comparative toxicity analysis of nicotine analog vaping products, inform regulatory decision-making, and lay the foundation for further toxicity testing. Utilizing advanced chemical and biological methods, this research aims to identify biomarkers of exposure and elucidate the potential health impacts of inhaling these new vaping products.

NIH Research Projects · FY 2026 · 2025-04

Project Summary/Abstract The health of female gametes (oocytes) decreases exponentially as maternal age increases causing infertility and miscarriage risks. Infertility affects at least 1 in 4 women of childbearing age in North America and more worldwide. While medical interventions including in vitro fertilization (IVF) have improved reproductive success, this is strongly dependent on gamete health. In a human female, oocyte production (oogenesis) begins before birth where two-thirds of precursor oocytes are normally eliminated by programmed cell death (apoptosis). The few germline cells that escape apoptosis mature into oocytes. Apoptosis affects oocyte quality through unknown mechanisms. Studies on the contribution of apoptosis to maternal fertility are limited due to challenges with performing experiments on humans and even mice, and an underrepresentation of women's health studies in science. Identifying mechanisms that promote oocyte health will enhance our understanding of factors leading to infertility and aid fertility health research. Organ structure is tightly coupled to function, and the ovaries where oocytes are made are no exception. Throughout animal phylogeny, oogenic germlines are syncytial; they comprise of large, elaborate cells wherein many nuclei reside in connected compartments. My preliminary work with the oogenic germline of the model animal Caenorhabditis elegans revealed that disrupting germline structure prevented apoptosis and decreased fertility. How apoptosis removes certain precursor oocytes at the expense of others is unknown. The goal of this study is to determine the role of apoptosis in oogenesis and oocyte health as a conserved mechanism like apoptosis may explain ovary autonomous mechanisms in place to control oocyte quality before ovulation. The experiments proposed will be performed using the oogenic germline of C. elegans, due to similarities in proportional time scales of age-related oocyte quality decline and syncytial nature of precursor oocytes within the ovary. C. elegans has profound experimental advantages including a simple regulation of apoptosis. The proposed work will be conducted under the mentorship of Dr. Amy Shaub Maddox, with valued collaborators and a vibrant scientific community. Dr. Maddox has extensive experience with C. elegans, contractility, cell shape regulation, advanced imaging methods, and theoretical modeling. The lab is well-equipped with all the tools necessary to define the interplay between apoptosis in germline architecture and function. This project provides a comprehensive training opportunity for me by teaching me animal physiology and cell biology, microscopy and data analysis, and mathematical modeling. I will obtain professional development through mentoring undergraduates, attending networking events, writing, and publishing my work, and presenting at cell biology conferences like the Triangle Cytoskeleton and the American Society for Cell Biology meetings, with the ultimate goal of becoming an independent scientist faculty member at a research university in the US.

NIH Research Projects · FY 2025 · 2025-04

Project Summary/Abstract The mental health of children in the United States is a national emergency, with notable and accelerating rates of anxiety, depression, and ADHD. Recent research suggests exposure to natural environments (green spaces) reduces risk for these disorders, alleviating stress, restoring emotional and physiologic resources, providing opportunities to build regulatory skills through risky play and physical activity, and reducing harm from environmental stressors, such as heat. The benefits of green spaces may be even greater in low-income areas, buffering or mitigating the impact of physical and social stressors, and serving to reduce mental health disparities. Green spaces offer the potential for novel, community-level support to offset risk for anxiety, depression, and ADHD, yet evidence-based guidelines on the timing and types of exposures that are beneficial are unclear, as well as the benefits of green spaces for children outside of urban areas. The paradox of high greenness but poor mental health in rural, low-income communities highlights the need to determine the factors that alter the relative impact of green spaces. The current proposal seeks to address these gaps, establishing 1.) the buffering role of green spaces on anxiety, depression, and ADHD risk in low-income, rural communities, 2) whether the relative benefits of green spaces are consistent across childhood, 3.) whether different types of green spaces (general vegetation density, proximity to public parks or residential exposure to woodlands, grasslands, or croplands) convey similar benefits, and 4.) whether green spaces buffer children from poverty-related adversity in these communities. Drawing from 18 years of observational, survey, and medical record data gathered from The Family Life Project, a population-based study of 1,292 children born in low-income, rural communities, the proposed project will derive new, remotely-sensed, geospatial measures of types of greenspaces around children’s residences, and integrate these measures with extensive child, family, and home data from 2 months to 16-18 years of age to address critical questions about the types and timing of green space exposures that offset risk for anxiety, depression, and ADHD. Findings could inform national policies and feasible interventions to protect and promote green space exposures in ways that maximize benefits for population-level mental health, and direct future research on the mechanisms through which exposure to nature offsets future mental health risk.