University Of Pittsburgh At Pittsburgh

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY High grade serous ovarian cancer (HGSOC) is the most common histologic subtype of ovarian cancer and remains a deadly malignancy facing women. Late-stage diagnosis, development of chemoresistance and high rates of recurrence all contribute to the low five-year survival rate of advanced stage patients. Thus, identifying novel drivers of tumor progression and heterogeneity continue to be high research priorities. We find that enhanced mitochondrial fusion driven by a splice variant of the mitochondrial fission protein Drp1/DNM1L is a phenotype of HGSOC. Loss of exon 16 in the Exon16/17 variable domain, denoted as Drp1(-/17), leads to Drp1 redistribution from mitochondria to microtubules (MTs), and consequentially to dampened fission, a fused mitochondrial network and enhanced mitochondrial metabolism. Importantly, Drp1(-/17) expression drives proliferation, metastasis and chemoresistance, and this is of clinical significance as Drp1(-/17) high HGSOC tumors are associated with poor patient outcome. In the present proposal we will test the hypothesis that the extra-mitochondrial function of Drp1(-/17) contributes to HGSOC tumor progression and heterogeneity. Our preliminary data suggest that microtubules are not simply reservoirs for Drp1(-/17) to drive mitochondrial fusion, but that Drp1(-/17) has localized microtubule functions associated with its interaction and recruitment of the multi amino acyl tRNA synthase complex (MSC) to centrosomes. Based on observations that Drp1(-/17) high tumors display greater aneuploidy and copy number alterations leads us to predict that localized centrosome protein synthesis facilitated by the MSC interaction with Drp1(-/17) at centrosomes leads to centrosome amplification and chromosome segregation defects. Intriguingly, many of the amino acids that are required for tRNA charging by the MSC are derived from the TCA cycle. Coincidentally, we find that Drp1(-/17) cells display increased levels of TCA cycle metabolites and downstream amino acid. This leads us to predict that increased mitochondrial fusion due to Drp1(-/17) microtubule association drives TCA cycle flux, and that this works in concert with the MT-dependent function of Drp1(-/17) by supplying amino acids for the MSC. We further propose that de novo amino acid synthesis provides survival advantages to Drp1(-/17) under amino acid deprivation and that this contributes to tumor progression. Our work is conceptually innovative as it investigates the unique interplay between mitochondrial morphology, amino acid metabolism, and localized regulation of protein synthesis at the centrosome to chromosome instability, a common feature of HGSOC. The results from this work will have major impacts on our understanding of HGSOC tumor heterogeneity. Moreover, our research is the first demonstration that a splice variant of Drp1 has unique extra-mitochondrial functions in a pathophysiologic context.

NIH Research Projects · FY 2026 · 2026-06

Project Summary: . The sex-specific cardiotoxic effects of doxorubicin (DOX), an anti-chemotherapeutic agent, are very well known. Adult premenopausal women have reduced cardiotoxic effects compared to age-matched men. The mechanisms underlying DOX cardiotoxicity have also been extensively investigated over decades. It is, therefore, surprising that the mechanisms that promote sex dimorphism in DOX cardiotoxicity is unknown and it is a gap in knowledge that will be addressed by this proposal. Metabolic impairment is a major upstream contributor of DOX cardiotoxicity where fatty acid (FA) metabolism is impaired by DOX. FA metabolism is regulated by the nuclear receptor peroxisome proliferator- activated receptor alpha (PPARα). Recent studies have shown that PPARα activation has different effects in male and female stroke outcomes and hypertrophy. In this proposal, we will test the role of PPARα in promoting sex dimorphism in DOX cardiotoxicity. In Aim 1, we will test the role of PPARα activation and FA metabolism in promoting sex-specific cardiotoxicity. In Aim 2, we will test the mechanistic pathways by which estrogen signaling promotes sex-specific regulation of PPARα. In Aim 3, we will test these cardioprotective therapies in tumor bearing mice and a novel human cardio-oncology organotypic platform. We will test the overarching hypothesis that estrogen mediated PPARα activation prevents FA metabolic impairment and ameliorates DOX cardiotoxicity. Preliminary results indicate that attenuated DOX cardiotoxicity in female mice was associated with increased PPARα activation, transcriptional activity and FA metabolism related markers. Genetic ablation of PPARα in mice abolished sex differences and worsened DOX cardiotoxicity. On the other hand, pharmacological PPARα activation in human cardiac organotypic slices ameliorated DOX cardiotoxicity. Cardiotoxicity will be assessed by multi-parametric physiological assessments. Cardiac metabolism will be measured by seahorse analysis; cardiac mechanics will be determined by echocardiography in mice and slice contraction measurement in human slices; cardiac electrophysiology will be assessed by electrocardiography. The interrelationship between metabolism, excitation, and contraction will be determined by applying our novel quadruple-parametric optical mapping technique to measure electrical excitation, contraction, and metabolic markers simultaneously. Modulation of cardiac structure under each of the above conditions will be assessed by histological and transmission electron microscopic analyses. Finally, molecular and transcriptional changes induced by PPARα activation during chemotherapy will be determined by western blotting, RNA sequencing and metabolomic/lipidomic analysis.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract A robust host immune response to inductive bioscaffolds is essential for their biodegradation. Specifically, peripheral macrophages are thought to be the key effector of this process, which is essential for tissue regeneration. The trafficking of immune cells to the bioscaffold nevertheless remains poorly understood, especially in the brain. It is hypothesized here that immune cells accumulate in the peri-lesional tissues prior to their invasion into the bioscaffold. The spatio-temporal dynamics of this invasion at the tissue/bioscaffold interface is crucial to understand and potentially control this process. Histological time course studies are limited in their ability to visualize this process, especially in tissue cavities with a diverse topology, such as a stroke. We therefore here propose to use 19F Magnetic Resonance (MR) based imaging techniques to non- invasively visualize the distribution of ECM hydrogel using perfluorotertbutyl-cyclohexane (PFTBC), as well as perfluorocrownether (PFCE) invading immune cells and more specifically macrophages during different phases of the immune response. This multi-color 19F MRI is unique in its ability to distinctly visualize the bioscaffold and the acute peripheral immune cell invasion in the same animal over time. We aim to: 1) visualize non- invasively the spatio-temporal dynamics of immune cells accumulation and invasion into hydrogels implanted into brain tissue cavities formed by stroke; 2) to timestamp different phases of the peripheral immune cell invasion by 19F MRI and 3) to monitor in vivo the ablation of immune cells and investigate their contribution to hydrogel biodegradation and tissue regeneration. The goal of this work is to improve our mechanistic understanding of immune cell trafficking between the host and bioscaffold. We expect that these studies will lead to novel methods to monitor and manipulate the contribution of peripheral immune cells to biodegradation and tissue restoration.

NIH Research Projects · FY 2026 · 2026-06

SUMMARY Chimeric Antigen Receptor (CAR) T cell therapy has made a significant impact on the treatment of hematological malignancies and is poised to be directed toward solid tumors, autoimmune diseases, and chronic viral infections. CAR T cells recognize target antigens on cell surfaces, activating immune responses that lead to target cell destruction. Recently, we developed a programmable universal CAR, termed SNAP-CAR, that can be directed to any cell surface antigen of interest via covalent binding with benzylguanine (BG)-modified adaptor antibodies. Universal CARs offer significant therapeutic advantages by allowing a single receptor to target multiple antigens through interchangeable adaptors, addressing a key challenge of resistance due to target loss or heterogeneity in cancer treatment. However, antibody adaptors present limitations, including high production costs, intravenous administration, and difficulty in deactivation if adverse effects arise, especially in multi-antigen targeting scenarios. Additionally, chemical modifications required to convert antibodies into homogenous, tumorspecific adaptors are complex. To overcome these limitations, we propose the development of cyclic peptidebased adaptors for universal CARs. Cyclic peptides offer several significant advantages over protein-based adaptors, such as ease of development and production, synthetic versatility, and potential for oral administration. Given their successful use in clinical imaging, cyclic peptides may also provide diagnostic benefits. We aim to design and synthesize several cyclic peptide adaptors to program SNAP-CAR T cells to recognize prominent cancer antigens. We will test these adaptors, and their combinatorial use, in vitro and in vivo using human tumor xenograft models. The proposed NSG mouse xenograft models are necessary to demonstrate anti-tumor efficacy in a physiologically relevant system that captures the in vivo interaction between SNAP-CAR T cells and cyclic peptide adaptors. This system encompasses critical parameters such as adaptor biodistribution, bioavailability, and stability, CAR T cell trafficking, tumor infiltration, and persistence, and potential off-tumor ontarget toxicity, which cannot be adequately recapitulated in in vitro systems. This approach represents a novel and versatile strategy for advancing universal CAR T cell therapy, with broad applicability in targeting multiple cancer antigens.

NIH Research Projects · FY 2026 · 2026-06

The Birch reduction remains largely non-chemoselective, limiting synthetic applications. Electrophilic additions to radical anion and anion intermediates of electron-rich monoarenes have also been found to be unfeasible. These shortcomings have persisted because the traditional Birch reduction conditions are not tunable. In 2021, we reported our Birch-type method using Li(0), ethylenediamine (EDA), and tBuOH in THF at 0–25 °C. We plan to develop unprecedented chemoselectivity for this Birch-type method to reduce one arene group over other arenes and even alkynes. Our preliminary work has indicated that the reactivity is tunable with various proton sources and amines. We will determine the structure-activity relationships of amines, substrates, and proton sources in order to predict optimal conditions for desired chemoselectivity. Moreover, this project will allow us to develop a silyl Birch-type reduction method to intercept the radical anion and anion intermediates, affording 3,6- disilylated 1,4-cyclohexadienes. The oxidative rearomatization of these products will generate anti-Friedel-Crafts products. The long-term goal of this study is to develop unprecedented chemodivergence and complexity- building strategies through Birch-type reductions to synthesize therapeutic agents. Aim 1 is to determine the roles of amines and proton sources in Birch-type reduction. The traditional Birch reduction proceeds through an electron transfer (ET) step to form a radical anion, followed by a proton transfer (PT) step to generate a dearomatized radical (ET-PT mechanism). We hypothesize that a proton-coupled electron transfer (PCET) involving π-hydrogen bonding, under our reaction conditions, is a viable alternative mechanism as the PCET process can bypass the higher-energy radical anion intermediate. We will investigate the mechanism experimentally and computationally to study the structures of Li-ligand complexes and the roles of proton donors. A mechanistic model from this aim will be used to rationally develop new chemoselective Birch- type reductions. Aim 2 is to develop chemodivergent Birch-type reductions. While chemoselective Birch(-type) reduction has not yet been developed, our preliminary studies have indicated that the judicious choice of proton sources in combination with the use of diamines besides EDA can achieve new chemoselectivity. Specifically, this aim will explore the chemodivergent Birch-type reduction of an arene in the presence of other arenes and alkynes. This aim will also inform Aim 1, which seeks to determine whether more acidic proton sources promote the PCET mechanism, selectively accelerating the reduction of more electron-rich arenes. Aim 3 is to develop Birch-type reduction-silylation with electron-rich monoarenes, previously impossible. This aim will overcome this barrier through the discovery of Li(0)-resistant electrophiles and non-nucleophilic amines to form solvated electrons. We recently determined that N-trimethylsilyl-imidazole met these requirements and plan to develop new synthetic methods, turning the Birch-type reaction into a complexity-building transformation, including a synthetic platform for anti-Friedel-Crafts products.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Available treatments for opioid use disorder (OUD) do not adequately address cravings and affective symptoms – factors that contribute to 60-90% of individuals with OUD experiencing relapse. Nicotine dependence is a major confounding factor limiting the translation of novel OUD treatments. Between 80-90% of individuals with OUD smoke. Unfortunately, smoking cessation treatments are not commonly provided during substance use, despite the fact that smoking cessation has positive outcomes for individuals with OUD. Despite the high prevalence of nicotine use in OUD, extremely few preclinical studies have assessed how nicotine exposure interacts with opioids’ long-term physiological effects. The studies outlined herein will examine effects of nicotine and opioids on the prefrontal cortex (PFC), an area involved in persistent cravings and anhedonia in OUD. In mice and humans, the mu opioid receptor is expressed by somatostatin-expressing inhibitory neurons (SST-INs), which receive inhibitory input from nicotinic receptor-expressing vasoactive intestinal peptide inhibitory neurons (VIP- INs). Our preliminary studies and the literature indicate that nicotine depolarizes VIP-INs and excites pyramidal cells through the VIP-IN ® SST-IN ® pyramidal cell disinhibitory motif. Thus, due to expression of MOR within SST-INs, and the expression of nicotinic receptors on presynaptic VIP-INs, PFC SST-INs are uniquely situated to undergo synergistic molecular and synaptic adaptations following combined exposure to nicotine and opioids. Furthermore, we present preliminary data that nicotine increases opioid use and decreases social approach during opioid withdrawal in mice. Based on this, we hypothesize that nicotine primes PFC SST-INs to promote opioid use and social anhedonia following opioid withdrawal. We will test this hypothesis through a series of controlled preclinical experiments and postmortem human brain studies. Aim 1 will determine whether SST cells in human PFC undergo pronounced transcriptional changes in individuals with OUD who use nicotine. Aim 2 will involve mechanistic studies in mice to determine the persistent adaptations to SST-INs elicited by combined exposure to nicotine and opioids. Aim 3 will determine whether modulating SST-IN activity can reduce polysubstance use and affective disturbances in withdrawal. These studies provide essential information on how polysubstance use of opioids and nicotine impacts the PFC. Results from these studies may have important ramifications for treatment development and will be critical for interpreting the extant preclinical literature that has largely excluded nicotine from study design.

NIH Research Projects · FY 2026 · 2026-06

Project Abstract The Doctor of Physical Therapy-PhD in Bioengineering Training Program at the University of Pittsburgh integrates outstanding evidence-based physical therapy education with innovative bioengineering research training to develop independent clinician-scientists who can identify clinically relevant questions that can be addressed by rigorous solutions grounded in bioengineering principles and who will be leaders in rehabilitation research. An established history of interdisciplinary collaboration between the departments provides the foundation for this program, which is structured to provide a synergistic environment between the clinical and research training so that the students have early engagement in research while they are studying physical therapy, as well as continued physical therapy skill development while they are immersed in their research training. The program will support up to 6 predoctoral trainees per year, and each trainee will be eligible for up to 3 years of support. They will be engaged in learning the fundamental principles of physical therapy, bioengineering, biostatistics, evidence-based practice, and applying the principles through closely-mentored laboratory training, manuscript preparation, and grant-writing. Training program specific activities that augment the individual DPT and PhD curricula include providing the trainees with research experiences during DPT phase, clinical experiences during the PhD phase, exposure to different career pathways, and regular training in the responsible conduct of research, methods to enhance rigor and reproducibility in research, and mentorship. The training program features clinical rehabilitation research faculty with expertise in varied populations (neurologic, orthopaedic, sports, pediatrics, geriatrics) and bioengineering research faculty with backgrounds in biomechanics, signal and image processing, neural engineering, machine learning) to provide a unique training program at the cutting edge of physical rehabilitation. The training program benefits from the institutional support of the University of Pittsburgh and its strong infrastructure of facilities, mentorship training, and resources for faculty and graduate student development.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Osmotic homeostasis is the most aggressively defended physiological setpoint in biology. This is because disruptions in cell volume alter essential physiological parameters, such as macromolecular crowding, ion concentrations, and membrane integrity, all of which are required for cellular function. Disruptions in cell volume are an important pathological feature of many acute and chronic human diseases, such as stroke, diabetes, hypertension, and kidney disease. Much of our understanding of the molecular mechanisms of osmoregulation is derived from cellular models, such as yeast and cultured mammalian cells. A major challenge to the field has been the difficulty in studying this process in a multicellular in vivo setting, where complex neuronal, endocrine, and extracellular matrix interactions, that are not present in cellular models, are preserved. Such questions can be addressed using a simple animal model system. C. elegans exhibits robust behavioral and physiological responses to osmotic stress. Understanding mechanisms of organismal osmoregulation in C. elegans will inform our understanding of human physiology and disease pathophysiology and could reveal novel methods to control pathogenic nematodes. The important and broad questions that we will address include: 1) How do lysosomes detect osmotic stress and signal specific gene expression programs? 2) What is the genetic architecture of pathways controlling organismal osmoregulatory physiology? 3) What are the interoceptive mechanisms that link changes in osmoregulatory physiological state to alterations in nervous system function? Over the last, 20 years, my lab has pioneered the study of osmotic homeostasis in C. elegans. Thanks to advances in CRISPR genome modification, whole-genome resequencing, automated behavioral analysis, and neuronal imaging and optogenetic stimulation, along with new molecular insights gained in the previous funding period, we are uniquely poised to define the integrative physiology of metazoan osmoregulation in unprecedented detail.

NIH Research Projects · FY 2026 · 2026-06

Uveitis is responsible for up to 10% of blindness in the US and is a leading cause of vision loss worldwide. Associated with diverse mechanisms of inflammation driven by a range of causes, diagnosis is challenging because it often relies on serological testing or systemic assessments rather than direct ocular measurements. Microglia and other retinal cells and circulating immune cells, as well as the cytokines they produce such as IFN- γ, TNF-α, IL-1, IL-6, and IL-12 influence uveitis pathophysiology as they shape the immune environment within the eye. However, there is a major gap in the understanding of how these key players of the immune response differ across the myriad forms of uveitis. Most of our knowledge comes from animal models due to a lack of tools for real time assessment of immune responses in the eye. We recently showed, for the first time, how adaptive optics ophthalmoscopy (AOO) methods can assess the immune response in posterior uveitis (PU), which affects the retina and/or choroid, in real time in the living eye. Our non-invasive, label-free approach offers promise to revolutionize the diagnostic toolkit for PU and may provide rigorous new biomarkers for evaluating treatment efficacy in the clinic and in clinical trials. It is known that the immune environment differs in infectious and non- infectious uveitis, so we will begin by defining these immunological differences quantitatively using conventional immunological assays including flow cytometry and cytokine analysis in a case-control study of well characterized patients (Aim 1). Next, we will test the hypothesis that these differences allow the use of AOO to detect morphological and dynamic changes of immune cells associated with the retina. Our preliminary findings suggest that AOO can also detect certain pathogens, and we will define the types of microbes that are causing infectious uveitis. We will also determine the other inflammatory biomarkers accessible to AOO in PU, such as vascular changes. Finally, in longitudinal study of a subset of patients, we will track the changes of these biomarkers over time in response to treatment. We predict that imaging will reveal distinct and significant differences between infectious and non-infectious uveitis, specifically: 1) the morphology and dynamic activity of immune cells moving in and through the retina, 2) the profiles of specific microbes related to infectious uveitis pathology, and 3) vascular and other microscopic retinal changes. To realize our goal of transforming the clinical toolkit for real-time monitoring of inflammation, we also need rigorous tools to generate quantitative metrics from the inflammatory biomarkers. We will develop these quantitative tools and generate new imaging protocols for uveitis (Aim 2). This will allow us to establish a novel analytical pipeline to analyze the retina in real-time to increase diagnostic efficiency and improve treatment monitoring. Together, the knowledge and tools developed here will lay the foundation needed to use these new technologies to improve patient care, including rapid diagnosis and real-time treatment monitoring.

NIH Research Projects · FY 2026 · 2026-06

Branched chain amino acids (BCAA) and their metabolites are critical molecules in modulating intermediary metabolism, serving as sensors to determine the switch between anabolic and catabolic processes. Almost two dozen BCAA inborn errors of metabolism (IEMs) have been identified and are the most common of the organic acidemias. The central hypothesis of this project is that the pathophysiology of disorders of BCAA metabolism is related to both BCAA metabolite imbalance and disruption of structural integrity of mitochondria related to the enzymes of BCAA metabolism. Our long-term goal is to characterize BCAA metabolism and its deficiencies and to develop novel therapies for them. Our group has previously demonstrated that led to profound derangements cellular homeostasis that exceeds the current limits of understanding of these disorders. This application has 3 specific aims. Specific Aim 1 is to demonstrate the interactions of the enzymes of BCAA metabolism in situ. We have developed a high resolution in situ cryo-electron tomography (cryo-ET) workflow to visualize the impacts of mitochondrial energy dysfunction on mitochondrial ultrastructure and electron transport chain structure. We propose extending this technology to visualize the in situ organization of BCAA metabolism enzymes in fibroblasts from patients with IEMs of BCAA. We hypothesize that proximal enzymes of BCAA metabolism functionally and structurally interact with respiratory chain supercomplexes, and that this interaction is disrupted in patient-derived cells. Specific Aim 2 is the development of a new therapy for propionic acidemia (PA) and methylmalonic acidemia (MMA). We have identified a novel compound (PMA010) that reduces pathogenic lysine propionylation in patient-derived cells from patients with propionic acidemia (PA) and normalizes protein succinylation, thereby improving cellular bioenergetics. We will use targeted metabolomics and high-resolution cryo-EM to determine changes induced by PMA010 on cells from PA and methylmalonic acidemia (MMA) patients, identify effects of PMA010 on mitochondrial protein lysine acylation and mitochondrial structure, and evaluate efficacy of PMA010 and/or analogs in vivo in a mouse model of PA. We hypothesize that these experiments will identify PMA010 as candidate for clinical trials for these disorders. Specific Aim 3 is to understand the physiology of the mitochondrial citrate transporter and its deficiency. The end products of BCAA catabolism enter the TCA cycle to make citrate. Human SLC25A1 encodes the mitochondrial citrate carrier (mCiC), which facilitates transport of citrate between the cytosol and mitochondrial matrix. Genetic defects in SLC25A1 lead to the organic acidemia combined D, L, 2-hydroxyclutaric acidemia (c2HGA). We will express and characterize the function 4 CiC isoforms, characterize their interactions with BCAA metabolic enzymes via cryo-EM, and use a multi-omics approach to assess the effects of SLC25A1 mutations on: (i) pathophysiology in c2HGA patient-derived fibroblasts. We will test phenylbutyrate (PB), a drug shown in our preliminary data to improve cellular bioenergetics and metabolite profile, as a therapy for this disease. We hypothesize that PB will demonstrate a therapeutic effect in cells and an SLC25A1 deficiency mouse model as pre-clinical proof-of-principle for patient clinical trials.

NIH Research Projects · FY 2026 · 2026-06

Individuals with chronic kidney disease (CKD) are often burdened with complex medication regimens, polypharmacy, and comorbidities, and thus commonly experience medication therapy problems (MTPs). MTPs are undesirable events experienced by patients that involve medication therapy and may result in suboptimal disease management, harm, or risk of harm; this includes omission of clinically indicated medications. For example, people with CKD often do not receive important guideline-recommended cardio-kidney protective medications. Up to 30-40% of US adults with CKD lack ACEi/ARB or statin therapy. Further, newer cardiokidney protective medications, such as SGL T2i or GLP1 RA, are widely underutilized in people with CKD, and nephrology-based prescribing of these therapies is particularly low. Collaboration with pharmacists in the nephrology clinic setting is a potential way to improve use of guideline-recommended cardio-kidney protective medications and address MTPs for individuals with CKD. The objectives of this proposal are to discover ways to optimize medication therapy in nephrology clinics, design and build a pharmacist-led population health management (PHM)-based intervention with end-user stakeholders, and test the intervention in a pilot trial. The specific aims are to: 1) Identify needs that would facilitate nephrology-based prescribing of guidelinerecommended cardio-kidney protective medications through stakeholder interviews; 2) Develop a stakeholderinformed intervention to address nephrology-based prescribing of guideline-recommended cardio-kidney protective medications for CKD; and 3) Test the feasibility and acceptability of the intervention in a pilot trial. Preliminary data generated from this research will support a future R-level application to evaluate pharmacistled interventions in a randomized clinical trial. This K23 award will support the training and career development of Melanie Weitman, PharmD. Dr. Weltman's career goal is to become an independent investigator who leverages pharmacy care practice to enhance CKD care, increase access to guideline-recommended medication therapy, and improve medication-related outcomes for individuals with CKD. Dr. Weitman will achieve this goal through support of her experienced mentoring team, which includes experts in population health management, pragmatic clinical trial design and implementation, and multidisciplinary care models. Specifically, this proposal will enable Dr. Weitman to achieve the following career development objectives: develop skills in 1) qualitative methodology and 2) human-centered design and intervention development; and 3) refine expertise in clinical trial design and data analysis. Dr. Weitman will also pursue overarching career development activities to strengthen leadership and academic contributions. Completion of these aims and objectives will facilitate Dr. Weltman's transition to becoming an independent investigator and leader in medication-related outcomes research in CKD.

NIH Research Projects · FY 2026 · 2026-05

Project Summary: Pancreatic ductal adenocarcinoma is notoriously resistant to chemotherapy and radiation therapy (RT). Lipid metabolism has been implicated in tumor cell intrinsic resistance to oxidative stress by protecting against ferroptosis, as well as cell extrinsic immunosuppression. Genes of de novo lipid production through lipogenesis are upregulated in many cancers including PDAC. This project proposes to characterize de novo tumor lipid production following cytotoxic therapy, as a means of escaping cell intrinsic cell death by evasion of ferroptosis, and immune mediated cell killing by promoting M2 macrophage polarization. We propose to test whether genetic and pharmacologic inhibition of lipogenesis alters the cellular and microenvironmental lipid makeup, and whether this alteration enhances the sensitivity to oxidative stress through ferroptosis, and immune evasion by ApoE mediated suppressive macrophage polarization. We believe this could represent a novel mechanism of therapeutic resistance that could be targeted with inhibitors of lipogenesis in combination with cytotoxic therapies already in clinical use. We propose to use in vitro and in vivo tumor studies, utilizing mass spectrometry, RNA seq, and flow cytometry to characterize the effects of cytotoxic therapy and inhibitors of lipogenesis on lipid makeup, immune infiltration and polarization, and tumor sensitivity to oxidative stress. Aim 1: Determine the impact of PDAC lipogenesis on tumor resistance to oxidative stress and cytotoxic therapy. We hypothesize that oncogenic KRAS upregulates lipogenesis proteins ACC and FASN in PDAC cells through SREBP1, leading to increased lipid turnover and ability to resist oxidative stress induced ferroptosis, driving therapeutic resistance. We will characterize the changes PDAC cellular lipid expression following oxidative stress by RT by mass spectrometry, and determine if inhibition of lipogenesis and Lands cycle abrogates these changes. We will then test if pharmacologic inhibition of lipogenesis and Lands cycle can sensitize PDAC cells to RT. Aim 2: Demonstrate the effect of PDAC lipogenesis on tumor ApoE expression, macrophage polarization and immune mediated therapeutic resistance to cytotoxic therapy in vivo. We hypothesize that RT increases PDAC lipid production and lipid oxidation by stimulating expression of SREBP1, leading to tumor immune suppression through ApoE secretion and M2 polarization, preventing anti-tumor immunity and immune clearance in the presence of cytotoxic therapy. We intend to test whether genetic and pharmacologic inhibition of lipogenesis lipid makeup of the tumor microenvironment after RT, altering ApoE expression and tumor macrophage polarization, and resistance to RT.

NIH Research Projects · FY 2026 · 2026-05

Abstract Oropouche virus (OROV) is an emerging Orthobunyavirus of concern to human health throughout the Americas. To date, OROV is the only known human arthropod-borne virus primarily transmitted by Culicoides midges. The recent upsurge in OROV infections driven by a novel viral reassortant, coupled with the geographical spread and the detection of more severe outcomes makes OROV an important threat to global health. Currently, there are significant limitations in our understanding of the epidemiology, ecology, and pathogenesis of OROV and its reassortants. This knowledge gap is particularly pronounced regarding vector-virus-host interactions that enable OROV to infect and disseminate in human skin, the critical organ for virus transmission. To address this gap, we will leverage a robust human skin model of arbovirus infection to define the cutaneous immunologic events during Culicoides probing and its impact on OROV replication and spread in human skin. This research proposal is in direct response to NIH Notice of Special Interest NOT-AI-24-048: “Immune Responses to Arthropod Feeding on Vertebrate Hosts”. Aim 1 will define the physiological effects of Culicoides probing in human skin immune response. Aim 2 will provide a comparative analysis between historical and contemporary OROV strains to pinpoint the contribution of Culicoides probing to cellular tropism, cutaneous immune responses, and viral dissemination. Our transdisciplinary and highly collaborative research team brings together unique strengths in cutaneous immunity of arboviruses (Priscila Da Silva Castanha), biology and ecology of Culicoides (Stacey Scroggs and Bethany McGregor), and bunyavirology (Cynthia McMillen). Our studies will address fundamental questions about the intricate interplay between the early host response to Culicoides probing and viral infection that are central to successful OROV transmission. This knowledge is pivotal for the development of therapeutic strategies and interventions against this neglected pathogen.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Spontaneous brain activity (i.e., resting state [RS]) is highly structured in both space and time. This salient feature of brain activity is universal across species. Nevertheless, we do not know how or why RS is structured. Despite this major knowledge gap, RS is routinely acquired with functional magnetic resonance imaging (fMRI) and the data is used for insights into brain organization in health and disease. Inferring human brain organization from rs-fMRI is a practice that has grown tremendously over the past 10 years; a trend bound to continue with the proliferation of MR scanners in the United States. But unless we improve our understanding of the neurobiological properties that confer structure onto RS, inferences from RS will not achieve their potential as a window into brain organization. This would jeopardize the impact of hundreds of studies per year and the massive investments from the NIH towards brain mapping (e.g., Human Connectome Project). Here, we propose a series of experiments to close critical knowledge gaps about the neurobiological properties of RS. Our objective is to shed light on the relationship between RS, cortical architecture, and cortical neurophysiology. We use a non- human primate model so that we can deploy high-resolution tools for recording RS, tracing cortical connections, and measuring neurophysiology. We conduct a large portion of our study in motor and somatosensory regions where we can use microelectrode mapping to accurately partition cortex according to function (e.g., hand control vs face control) and cortical area borders (e.g., motor vs premotor cortex). Towards our objective, we propose three Specific Aims. Aim 1 benchmarks cortical parcellation inferred from RS against ground truth cortical divisions. We record RS with fMRI and intrinsic signal optical imaging (rs-ISOI), which is operationally like fMRI but provides higher contrast and spatial resolution. We leverage the statistical dependencies in the recorded time series to generate high resolution maps of cortical networks. The spatial organization of those maps is then quantified by measuring their overlap with anatomical and functional divisions of cortex. Aim 2 benchmarks functional connectivity (rsFC) inferred from rs-ISOI and rs-fMRI against ground truth neuroanatomical connections. rsFC is mapped for sites throughout sensorimotor cortex. We directly compare those connectivity maps to the anatomical maps that we reveal from the same sites using tracers, microstimulation, and fiber tractography. Aim 3 investigates the neurophysiological basis of RS. We place electrode arrays throughout cortical networks and record time series of neurophysiology, rs-ISOI, and rs-fMRI. We then measure the extent of co-fluctuations between the neurophysiology and imaging time series. Our proposed multi-modal approach will shed light on the neurobiology of RS. We will therefore serve vast segments of the neuroscience community that leverage RS. Knowledge gained here will set the stage for next generation connectome projects, which will annotate cortical architecture at the level of cell types, receptors, and genes. Such a resource would transform how our field approaches the functional organization and adaptive rewiring of cortical networks.

NIH Research Projects · FY 2026 · 2026-05

We are requesting funds to purchase a single integrated system. A 4 color confocal microscope with resonant scanning capabilities on a motorized inverted stand with four GaSP detectors, two with customizable spectral tuning. This instrument will be housed within the Center for Biologic Imaging (CBI) at the University of Pittsburgh School of Medicine. The mandate of this core facility is to provide access to a full range of light and electron optical, image analysis, and morphometric methods to all research groups within the University of Pittsburgh School of Medicine. Point scanning confocal microscopy is an essential service provided by the center, and until last fall the core provided access to 6 different systems to our user base (>300 research teams and @1000 individual users). The average usage time for each of these systems is between 2,500 and 3000 hours a year and almost all the use is by groups supported by NIH grants. In September of last year the oldest of these systems, a 19 year old FV1000 (1S10RR022637-01) had multiple failures in the laser combiner which could not be repaired due to a lack of parts (see letter) and the microscope was retired. This January we were told that the oldest of our Nikon A1 confocals (16 years old) had multiple components which would no longer be covered by Nikon service due to a lack of parts (gas lasers and tubes and other hardware) (see letter) which means that future failure may make the system non-repairable and it should be replaced. The NIH funded research base in our institution continues to grow, and given the current use levels we feel our confocal capabilities are at saturation in fact the lack of available instrument time has become a major factor limiting the utility of the center, which in turn which has lead to considerable frustration amongst users. As pointed out by major users the requested instrument offers multiple technical advances over our current systems which motivate the specific instrument choice. However, beyond improved technology the absolute primary reason for submitting this application is to provide critically needed contemporary instrument time to a growing body of NIH supported users of the imaging core here at the University of Pittsburgh.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY This R21 application is in response to the NIH Notice of Special Interest (NOSI, NOT-DA-24-012) to understand xylazine misuse and consequences. Co-use of xylazine with opioids (frequently fentanyl) has become a major threat to human health in the United States. No FDA-approved medications, including the opioid overdose reversal medication naloxone, have efficacy in reversing xylazine’s effects. Although xylazine is a known non- selective agonist of α2-adrenergic receptors (α2-ARs), none of the α2-AR antagonists can effectively reverse all deleterious effects induced by xylazine, from central nervous system depression to severe skin lesions and infection, suggesting that most harmful manifestations of xylazine probably result from xylazine’s action on receptors other than α2-AR. The goal of this application is to fill a critical knowledge gap in understanding the pharmacological and pathological impact of xylazine and identify effective compounds that are proven "safe for human use" and can offset xylazine-caused damage. On which major receptors other than α2-AR does xylazine act? What are the functional consequences and immune responses due to xylazine’s actions? What compounds can effectively reverse damage induced by xylazine? We strive to answer these questions. Our preliminary studies show that xylazine inhibits α7 nicotinic acetylcholine receptors (α7nAChRs) and the inhibition can be offset by a class of natural products. Inhibiting Ca2+-conducting α7nAChRs is known to produce a broad spectrum of negative impacts because of the widespread expression of α7nAChRs across various cells in the human body. α7nAChRs play an important role in regulating the central and peripheral nervous systems as well as the cholinergic anti-inflammatory pathway. Here, we will investigate xylazine-induced functional changes of α7nAChR in different types of human cells and identify natural products that can effectively reverse the xylazine- induced functional consequences. We will also investigate xylazine-triggered cell death and inflammatory responses and determine how effectively the identified natural products reverse xylazine-caused damage. The influence of fentanyl on xylazine’s effects will be investigated to provide much needed insight into complications from xylazine-opioid misuse. Cell types to be investigated include human neural and skin cells that natively express α7nAChRs and opioid receptors and are clinically relevant to the organs and tissues linked to the deleterious effects of xylazine. The outcomes from the proposed studies will broaden the current understanding of xylazine misuse and lead to potential treatment strategies for rapid therapeutic deployment to xylazine/opioid users.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Reproductive-age youth with epilepsy (RAYE) are at risk for adverse reproductive health outcomes. Certain antiseizure medications (ASMs) reduce the efficacy of some contraceptives and some ASMs adversely affect fetal development. Unplanned compared to planned pregnancy among people with epilepsy has been associated with increased rates of pre-term birth, congenital malformations, and seizures during pregnancy. Information about planning pregnancy, including use of contraception, is especially crucial for youth, who are at higher risk of unplanned pregnancy than adults. RAYE are also more than twice as likely to be prescribed teratogenic ASMs as adults over 24 years old. As reproductive health education is known to lead to better reproductive health outcomes for adolescents, informing RAYE about reproductive health and epilepsy is imperative. An important strategy to educate RAYE about epilepsy and reproductive health is through child neurology clinicians. My prior work found that RAYE and caregivers want epilepsy-related reproductive health counseling from child neurology clinicians, as is recommended by the American Academy of Neurology and Child Neurology Foundation. Yet my research has indicated that child neurology clinicians often omit this counseling or miscommunicate key messages. To address these challenges, I have developed a prototype of a multi-level intervention: Counseling Adolescents about Reproductive Health and Epilepsy (CARE). CARE combines 1) child neurology clinician online counseling skills training, 2) follow- up Short Messaging Service (SMS) messages for RAYE, and 3) SMS messages for caregivers to empower them to reinforce healthy behaviors. Through this proposed project, I will 1. optimize CARE with input from key stakeholders (clinicians, RAYE, and caregivers), 2. test the feasibility of CARE through a pilot trial, and 3. assess determinants of implementation of CARE through follow-up qualitative interviews with trial participants (clinicians, RAYE, and caregivers). These aims will provide the foundation for a future R01 application for a multi- center cluster-randomized controlled trial to test the clinical efficacy of CARE. This K23 proposal will support career development objectives in engaging adolescents in research, intervention development, conducting clinical trials, and implementation science. This grant will prepare me to become an independent investigator focused on optimizing the reproductive health of RAYE.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY Human metapneumovirus (HMPV) is a major cause of acute respiratory disease worldwide and the second most common cause of lower respiratory infection in US children. There is no targeted treatment for HMPV, and so it is not routinely tested for in clinical settings. HMPV mutates rapidly, so it can evolve variants that can become the predominant circulating strain in a population. However, HMPV is not typically under genomic surveillance, so the genomic factors that propel certain strains to become dominant remain undefined. Understanding which mutations are associated with dominance would greatly help the development of preventive and therapeutic treatments. My research project will be the largest prospective population-based HMPV genomic epidemiologic study. Using a large and robust collection of HMPV-positive clinical samples collected with corresponding patient data from 7 US cities, this study aims to decipher the genomic factors that contribute to strain dominance and to determine the extent to which viruses in different HMPV genomic subgroups differentially circulate in the population and are associated with disease severity. My long-term career goal is to become an independent investigator in infectious diseases with an initial focus on understanding the genetic mutations that enable dominance of a particular HMPV strain. My research project and career development activities will provide critical training for me to gain expertise in (1) viral genomic sequencing and genomic analysis tools to study the evolution of HMPV and (2) epidemiological methods to study patient and viral factors associated with infection and clinical outcomes.

NIH Research Projects · FY 2026 · 2026-05

Project Summary / Abstract Type 2 diabetes (T2D) affects nearly half a billion people worldwide and caused 6.7 million deaths in 2021 alone. While high blood sugar is a hallmark of the disease, researchers are increasingly focused on identifying what causes the disease to develop in the first place—especially at earlier, more treatable stages. In our recent study, we discovered that problems with a type of fat called sphingolipids may be one of the earliest warning signs of T2D. These fats are important for maintaining healthy cell membranes and signaling. Using advanced tools to analyze blood samples, we found that people who went on to develop T2D had significantly lower levels of sphingolipids. We then developed a new software tool, called metGWAS 1.0, which helped us link these changes to a specific gene called CERS2. CERS2 plays a key role in producing sphingolipids. One particular genetic variant of this gene, known as rs267738, is found in about 7% of people and causes a 33% reduction in CERS2 function. Although this gene variant is common and strongly linked to T2D in large genetic studies, no one had proven whether it actually causes the disease—until now. Our research shows that loss of CERS2 function can directly lead to T2D-like symptoms, even in the absence of obesity or other typical risk factors. In both mice and human cell models, we observed that when CERS2 was not working properly, the pancreas secreted less insulin (the hormone that lowers blood sugar), and the liver showed signs of metabolic dysfunction, such as high cholesterol and liver damage. We also found that this dysfunction is connected to two key molecular pathways—called mTORC1 and PPAR—which are known to be important in maintaining normal metabolism in the pancreas and liver. Ultimately, we hope this work will uncover new ways to treat or prevent T2D—by targeting the root causes of the disease, not just its symptoms.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Hearing loss is the most common sensory deficit and often arises from dysfunction of cochlear hair cells. Hair cells express precisely localized protein complexes, including the mechanotransduction (MET) channel, which converts mechanical sound waves into electrical signals and is essential for hearing. Although this heteromeric protein complex is critical for auditory function, significant knowledge gaps remain regarding how the MET channel components are assembled, trafficked, and localized within hair cells. Proper trafficking of MET proteins—from the endoplasmic reticulum (ER) to the stereocilia via the apical cell body—is essential for hearing. Many forms of hereditary hearing loss are linked to deficits in MET protein localization, underscoring the importance of understanding the underlying molecular mechanisms. TMC1 and TMC2 are the pore-forming subunits of the MET channel, with TMC1 being the predominant form during most of life. Our recent work identified Transmembrane O-methyltransferase (TOMT) as a critical regulator of TMC1/2 localization, MET channel activity, and auditory function. TOMT is the first known molecule that regulates MET protein localization without being part of the MET complex itself. While TOMT binds TMC1, it is excluded from the stereocilia and instead localizes to the ER and cytoplasm within the hair cell body. Importantly, TOMT alone is insufficient to promote TMC1 ER exit in non-hair cells, suggesting the need for additional, hair-cell-specific factors. We identified novel molecular chaperones—PEX3, PEX16, and PEX19 (collectively, PEX proteins)—that are expressed in hair cells and interact with TOMT to enhance TMC1 stability. Mutations in PEX genes cause hearing loss and peroxisomal biogenesis disorders in humans and mice. Based on these findings, our central hypothesis is that trafficking and localization of TMC1 is regulated by a specialized, hair-cell-specific machinery involving TOMT and PEX proteins. To test this hypothesis, we will 1) define how TOMT regulates TMC1 trafficking and stereocilia localization by mapping TOMT-TMC1 interaction domains and examining the effects of deafness-linked mutations on subcellular localization and auditory function; and 2) generate and employ knock-in and conditional knockout mouse models to characterize expression patterns and manipulate inner ear gene function of PEX proteins, assessing their individual and synergistic roles with TOMT in regulating TMC1 stability, trafficking, and auditory function. This project will establish a new conceptual framework for understanding how hair cells coordinate the trafficking and localization of essential auditory proteins including TMC1. Our findings will reveal molecular checkpoints and potential therapeutic targets for hearing loss associated with defects in protein localization, including mutations in TOMT and PEX genes.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT This project aims to develop a generalizable computational platform that integrates systemic omics, imaging, and genetic data to model early, anatomically localized tissue vulnerability across major organ systems. While traditional biomarkers are widely used in clinical care, they lack regional specificity and mechanistic interpretability, limiting their ability to forecast subclinical dysfunction or guide targeted interventions. This research will deliver a flexible platform that unites early spatial risk prediction, individualized molecular mechanism inference, and therapeutic discovery to support precision health across the brain, heart, lungs, liver, kidneys, pancreas, muscle, and beyond. The platform will link systemic molecular profiles, proteomics, metabolomics, and transcriptomics where available, to spatially resolved imaging-derived phenotypes (IDPs), such as brain connectivity, cardiac strain, or organ-specific tissue composition. It will model genotype–biomarker–IDP interactions to stratify individual-level susceptibility, leveraging polygenic risk scores and common variants that act through intermediate molecular traits. The framework will support therapeutic discovery through a deep learning–based drug repurposing tool that identifies candidate interventions capable of reversing early molecular dysfunction, prioritizing targets linked to inflammation, metabolism, fibrosis, and mitochondrial stress. To ensure broad applicability, the project will harmonize multimodal data across cohorts and organ systems and disseminate standardized pipelines and prediction tools.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Celiac disease (CeD) is a chronic immune-mediated disorder triggered by gluten ingestion in genetically susceptible individuals and currently lacks a curative treatment. The only available therapy is a lifelong gluten- free diet (GFD), which is burdensome and frequently ineffective, leaving 40-60% of patients symptomatic, underscoring the urgent need for alternative therapeutic strategies. CeD is marked by a loss of oral tolerance (LOT) to gluten, which precedes disease pathology. In genetically susceptible hosts, LOT is driven by proinflammatory dendritic cells (DCs) that promote proinflammatory T helper 1 (Th1) responses to dietary gluten, rather than tolerogenic regulatory T cell (Treg) responses. While HLA-DQ2 or DQ8 alleles are necessary for disease development, they are not sufficient, pointing to the role of environmental factors such as viruses and commensal gut microbes in shaping oral tolerance. We have gained significant insights into the mechanisms of how microbes’ impact oral tolerance. Our prior work showed that enteric infection with reovirus promote LOT to gluten, while colonization with a gut commensal protist protects against this effect. We also make use of MHC-II-restricted gluten tetramers that allow us to track gluten-specific T cell responses under physiologically relevant conditions. Recently, several independent laboratories provided strong evidence that DCs expressing the transcription factor RORgt (RORgt+ DCs) are essential for promoting oral tolerance by promoting Treg responses to dietary model antigen ovalbumin. This suggests that the effects of viruses and protists on oral tolerance may, at least in part, be modulated through RORgt+ DCs. Using genetic models to selectively and inducible deplete RORgt+ antigen-presenting cells and cutting edge tools to track gluten-specific T cell responses, we will: (1) test the requirement for RORgt+ DCs in regulating virus- induced LOT, restoring oral tolerance, and mediating protist dependent protection against virus-induced LOT and (2) determine how virus infection and protist colonization transcriptionally and functionally modulate RORgt+ DCs. Understanding how microbes modulate tolerogenic DCs will illuminate novel mechanisms of CeD pathogenesis and identify therapeutic targets for restoring oral tolerance to gluten in patients with CeD.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Across species, reproduction and immunity compete for limited physiological resources—a trade-off that becomes increasingly consequential with age, as both reproductive capacity and immune competence decline. In mammals, proteins that mediate these relationships to coordinate reproductive investment, immune regulation, and somatic maintenance are scarcely defined. This presents a critical gap in our understanding of reproductive aging and age-related disease susceptibility. This proposal investigates whether the C. elegans protein TCER-1, an established regulator of the fertility–immunity–longevity axis, has a conserved functional homolog in mammals. We previously identified TCER-1 as a pro-longevity factor that maintains fertility with age by suppressing immune and stress responses. Its homolog in Arabidopsis also promotes fertility and represses immunity, and our collaborative studies reveal a similar role for the Drosophila TCER-1 in repressing stress resistance, highlighting conserved trade-off regulation across kingdoms. TCER-1 is homologous to the mammalian transcription elongation and splicing factor, TCERG1, which has been linked to human disease in GWAS studies, though it’s in vivo function remains uncharacterized. This proposal is based on our preliminary data showing that (i) worm and mammalian TCERG1 both regulate alternative splicing of immune-related genes, and importantly (ii) mouse and human TCERG1 are enriched in germ-cells and exhibit a major age-related decline in female oocytes. In this exploratory proposal, we aim to assess whether human or mouse TCERG1 can functionally replace TCER-1 in C. elegans (Aim 1) and to define the role of mouse TCERG1 in fertility, ovarian reserve and reproductive aging by using existing and newly generated knockout models (Aim 2). By uncovering a potentially conserved molecular regulator of physiological trade-offs, this project aims to illuminate how reproductive aging shapes immune function and broader somatic aging, and to lay the groundwork for future interventions that support reproductive health in aging individuals.

NIH Research Projects · FY 2026 · 2026-05

Patients with diabetes mellitus (type 1 or 2) have a total lifetime risk of a diabetic foot ulcer (DFU) complication as high as 25%; 14-24% of them suffer from amputation. People with T1D develop DFU at a younger age and are at a much greater risk of amputation and hospitalization secondary to a DFU compared to T2D. The difference between pathophysiology and outcomes for individuals with T1D versus T2D is poorly understood and understudied. Family and twin-based studies have identified significant genetic components especially single nucleotide variations (SNV) in T1D as compared to T2D subjects. However, systematic patient-based genetic studies of T1D DFU are scanty, and the proposed work is aimed at seeding a novel paradigm in wound healing research. The originality and strength of our study stems from the genome-wide genotyping feasibility studies on robust quality controlled and parametrically qualified genotyped data of 149 chronic wound patients with diabetes status. This study identified 20576 SNV significantly associated with human chronic wounds (p- value<0.01, CR>97%, MAF>0.01). Majority (>60%) of these SNP were predicted to be causative for truncated or nonfunctional proteins using Variant Effector Prediction analysis were identified. To investigate the clinical significance of wound associated SNV, a meta-analysis against the phenotypes annotated in GWAS catalog was conducted as reported. These SNVs were intersected with manually curated >270,000 GWAS SNPs annotated with ~900 GWAS phenotypes collected from ~2500 studies. Enrichment analysis of the above intersected SNVs was performed against these GWAS phenotypes and respective odds ratio, and level of significance were calculated using Fisher’s exact test. These analyses identified “obesity” as the most significantly enriched GWAS-phenotype (log2 odds ratio = 4.06, p-value= 4.94E-12) for wound associated SNPs predominantly present in fat mass and obesity-associated (FTO) gene. This proposal is responsive to RFA-DK-26-009 for the New Investigator Gateway Award for collaborative type 1 diabetes (T1D) Research through Diabetic Foot Consortium (DFC). The objective of the proposed work is to determine SNV T1D and T2D that contribute to diabetic wound closure. This study will investigate the wound tissue already collected from patients with open DFU (N=50 with T1D and n=100 T2D) enrolled in the DFC Master Protocol. The following specific aims are proposed: 1.0 Aim 1. Identify SNV uniquely associated with T1D non-healing phenotype. T1D vs T2D will identify T1D-specific SNV (SNVT1D). Healing vs non-healing will identify SNVT1D-NH. SNVT1D-NH will be shortlisted to obtain candidate SNVT1D- NH (cSNVT1D-NH) based on overlap with obesity-associated SNP. 2.0 Aim 2. Test the functional significance of cSNVT1D-NH in wound healing mechanisms in vitro. Gene editing to specifically induce risk to non-risk alleles of specific cSNVT1D-NH using CRISPR/Cas9 genome editing improves: 2.1 epidermal keratinocyte migration in an in vitro scratch model; 2.2 formation of well-perfused and non-leaky vessels by microvascular endothelial cells using 3D-angiogenesis assay; and 2.3 augmentation of collagen deposition and maturation by dermal fibroblasts.

NIH Research Projects · FY 2026 · 2026-05

Koob and colleagues have postulated that an imbalance between neurotransmitters in the brain stress and anti-stress systems drives negative reinforcement and compulsive alcohol use in alcohol use disorder (AUD). Nociceptin (N/OFQ), which binds to the nociceptive opioid peptide receptors (NOP), is one such neurotransmitter in the brain that regulates stress and resilience in animal models of addiction. N/OFQ, when infused in the brain, increases corticosterone, adrenocorticotropic hormone, and corticotrophin- releasing factor (CRF), all components of the hypothalamic-pituitary-adrenal axis that regulate stress responses. CRF infusions have also been shown to upregulate NOP, presumably to enhance N/OFQ signaling, in brain regions that regulate stress. Surprisingly, both NOP agonists and antagonists show therapeutic effects in rodent models of AUD. However, rodent models do not clarify which, if any, subtype of AUD subjects (e.g., heavy drinking) will benefit from treatment with NOP agonists Vs. antagonists. Previous [11C]NOP-1A PET studies conducted by our group in humans with AUD have demonstrated lower binding (VT) to NOP receptors in heavy relative to nonheavy drinking AUD subjects. In these AUD PET studies, lower NOP receptors also predicted relapse to alcohol in a 12-week contingency management protocol that incentivized subjects with money to abstain. Assuming lower NOP receptors reflect higher N/OFQ levels; these results suggest that increased N/OFQ signaling in heavy drinkers contributes to their inability to abstain during treatment. Blocking excessive N/OFQ signaling with a NOP antagonist drug (LY2940094) has also been shown to decrease heavy drinking days and increase abstinent days in AUD subjects in clinical trials. An emerging view in the field is that a hyperactive N/OFQ-NOP receptor system, in response to increases in CRF transmission, induces hyperkatifeia, negative reinforcement, and relapse in AUD. To investigate CRF X NOP interactions in humans, we designed a PET experiment to measure [11C]NOP-1A VT before and after an intravenous (IV) hydrocortisone challenge in healthy controls (HC). Hydrocortisone administration led to a 10 to 15% increase in [11C]NOP-1A VT in brain regions, including the amygdala. Increased NOP measured in response to cortisol, and by extension, CRF, in this paradigm reflects an individual’s ability to enhance N/OFQ transmission during stress. Here, we propose to use this novel imaging paradigm to compare hydrocortisone-induced increases in [11C]NOP-1A binding (DVT) in the amygdala (and secondary reward regions) in heavy drinking AUD subjects Vs. HC. We hypothesize that hydrocortisone-induced increases in [11C]NOP-1A binding (DVT) will be larger in heavy drinking AUD relative to HC (aim 1), and this will predict relapse to alcohol (aim 2). Such a result will support the presence of a hyperactive NOP receptor system in response to increases in cortisol/CRF during conditions such as stress, chronic pain, etc., promoting relapse in heavy drinking AUD subjects.