University Of Oklahoma Hlth Sciences Ctr

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT The neurotransmitter acetylcholine regulates a diverse range of physiological functions and it is known that immune cells can respond to and synthesize acetylcholine in response to viral infections. Within the influenza A virus (IAV) infected lung, T cells are recruited to combat the infection. Upon an IAV infection activated T cells induce the gene choline acetyltransferase (chat) which results in the production of acetylcholine. I have found that aged mice are less likely to survive IAV infection and have fewer chat+ IAV-specific T cells than young mice. However, it is unclear why this decrease happens in aging mice. Humans over the age of 65 become hyper susceptible to IAV infection, while this is traditionally attributed to the changes in the immune landscape and quiescence of the immune system, little is known if there are other contributing factors. Using a mouse model wherein chat is not expressed in T cells I found that young mice that cannot produce T cell-derived acetylcholine have decreased survival and defective recovery from IAV infection. However, these mice clear IAV normally and have no differences in T cell infiltration into the lungs. Instead, I found that inflammation in the lungs is sustained well after viral clearance when T cells cannot produce acetylcholine. While this inflammation could be driven by many immune cell types, the only significantly elevated immune cell type in the lungs are neutrophils. Neutrophil depletion in vivo in the mice unable to synthesize T cell-derived acetylcholine rescues recovery. Additionally, the in vitro presence of acetylcholine prevents neutrophil migration toward a chemoattractant. This observation indicates a direct effect of T cell-derived acetylcholine on neutrophils. Taken together, these results suggest that acetylcholine-producing T cells are a critical source of acetylcholine in the lung during recovery from influenza infection. This proposal seeks to understand if (1) epigenetic changes are responsible for the decrease in chat expression in aged mice, (2) inhibiting the breakdown of acetylcholine can rescue weight recovery, and (3) how neutrophils are being directly impacted by T cell-derived acetylcholine. Based on my preliminary studies I hypothesize that acetylcholine plays a central role in recovery from viral infection. This hypothesis will be tested in the following aims.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Metabolic-associated steatotic liver disease (MASLD) is the most prevalent chronic liver disease affecting adults and up to 40% of children with obesity. A growing body of evidence supports the role of early life stressors in the etiology of MASLD. In humans and animal models, exposure to maternal Western diet (mWD) consumption and obesity during gestation and lactation increases offspring risk for steatosis and more severe metabolic- associated steatohepatitis (MASH). MASH involves the recruitment of bone marrow derived monocytes to the liver, which give rise to macrophages (Mφ) that are unable to resolve inflammation or repair damage, accelerating liver fibrosis. Previous studies show mWD skews bone marrow (BM) derived Mφ (BMDMs) toward a proinflammatory phenotype by remodeling fetal hematopoietic stem and progenitor cells (HSPCs). Importantly, HSPC programming persists long-term, as BMDMs from juvenile offspring exposed to mWD in early life are similarly primed for inflammation by an as-yet characterized mechanism that drives inflammation long term and may contribute to ongoing fibrosis in the juvenile liver. Mitochondrial dysfunction, oxidative stress, and inflammation are hallmarks of Mφ trained immunity but crosstalk between these processes due to mWD and their impact on MASLD are not well understood. Our preliminary data shows that mononuclear cells (MNCs) from BM of 3-week-old mice exposed to mWD have decreased oxidative capacity relative to MNCs from BM of control offspring from chow-fed dams. Therefore, we hypothesize that mWD alters mitochondrial function in HSPCs during critical windows of early development to promote BMDM activation and hepatic inflammation, thereby increasing susceptibility to MASH. The overall goal of this project is to understand the mechanisms for how mitochondria are maladaptive to maternal WD during gestation and/or lactation and how specific pathways contribute to the pathogenesis of inflammation through HSPC remodeling of BMDMs. This fellowship has two aims: 1) determine the impact of mWD exposure during pregnancy and lactation on HSPC and BMDM mitochondrial metabolism in mice at weaning and 2) determine the long-term effect of mWD during gestation or lactation on hepatic Mφ populations, Mφ function, and liver fibrosis in adult offspring following WD challenge. Our approach utilizes metabolomics and proteomics, fluorometry, and respirometry to assess mitochondrial physiology and single cell RNA-sequencing to identify unique populations of hepatic Mφ, capacity for liver repair, and fibrosis in livers of adult mice from the same early life mWD exposures. Completion of this work will provide me with training in mouse research, immune cell metabolism, deeper analysis of mitochondrial physiology, bioinformatics, and use of ‘Omics technologies to use in my next steps in career development. Impact: the work from this fellowship will fill a gap in our understanding of maternal diet’s impact on the pathogenesis of pediatric MASLD.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY/ABSTRACT Adolescent nicotine vaping remains prevalent alongside increases in cannabis vaping, nicotine-cannabis co-vaping, and daily/near-daily nicotine and cannabis vaping rates since 2019. Nicotine and cannabis vaping are associated with nicotine and cannabis dependence, carcinogen exposure, and pulmonary, cognitive, and mental health symptoms, with co-vaping posing additive health risk. Despite the critical role of parents in preventing adolescents’ tobacco and cannabis smoking, adolescents’ vaping poses difficulties for parents in their ability to engage in vaping prevention parenting behaviors (VPPBs), as vape devices may be disguised as other products and modified to vape cannabis and higher levels of nicotine. Less than half of parents report knowledge of vape device characteristics, vaping health consequences, adolescent vaping norms, and youth-targeted vaping marketing. Social media messages about vaping and VPPBs are ideal prevention tools, as 90% of US parents use social media regularly including for health information and parenting advice. This proposal is informed by social cognitive theory and the health belief model and builds on our team’s expertise in longitudinal studies of parenting and adolescent substance use and experimental studies testing youth vaping messaging. The overall goal of this work is to understand how to empower parents to prevent adolescent nicotine and cannabis vaping. We will conduct a 6-wave longitudinal survey study among 1,000 US parent-adolescent (ages 12-17) dyads recruited via social media and a 5-week online experimental study among a subset of 300 parents with low parent vaping knowledge (PVK) to test messages designed to improve PVK and VPPBs. Parents will be randomly assigned to view Facebook posts in a 4 (within-persons) X 3 (between-persons) mixed design. Each parent will view: 1) 4 sets of messages aimed at promoting PVK (device characteristics, harms, norms, youth-targeted marketing) in a randomized order; and 2) one of 3 sets of messages aimed at promoting VPPBs (rules, monitoring, communication). Experimental outcomes include PVK (i.e., device characteristics, health consequences, vaping norms, youth-targeted marketing), use of VPPBs (i.e., rules, monitoring, communication), and adolescents’ vaping outcomes (i.e., any current nicotine/cannabis vaping, frequency of nicotine/cannabis vaping, co-vaping vs. nicotine-/cannabis-only vaping, susceptibility to future nicotine/cannabis vaping). We will address 2 specific aims: Aim 1 (longitudinal surveys) - Examine 1a) direct associations of PVK and VPPBs with adolescents’ nicotine and cannabis vaping trajectories; and 1b) indirect associations of PVK with adolescents’ nicotine and cannabis vaping trajectories via VPPBs as mediators; and Aim 2 (experimental study) - Examine: 2a) within-persons effects of PVK messaging themes on PVK outcomes and between-persons effects of VPPB messaging themes on VPPB outcomes; and 2b) within-persons effects of PVK messaging themes on adolescents’ vaping outcomes via increases in PVK and between-persons effects of VPPB messaging themes on adolescents’ vaping outcomes via increases in VPPB use. This proposal is responsive to NIDA’s NOT-DA-22-004: Epidemiology of Drug Abuse and will inform large-scale social media messaging campaigns empowering parents to prevent adolescent nicotine and cannabis vaping initiation and progression.

NIH Research Projects · FY 2026 · 2026-05

PROJECT ABSTRACT. Prostate cancer (PCa) is the most commonly diagnosed cancer among Black men in Oklahoma and the U.S., with mortality rates 2 to 4 times higher than other racial groups. Despite evidence that prostate-specific antigen (PSA) screening reduces mortality and healthcare costs, screening rates remain low, particularly in Oklahoma (27% screened in the past two years). Barriers such as limited provider engagement, knowledge gaps, and healthcare access challenges contribute to lower screening rates, underscoring the need for innovative, scalable interventions to improve early detection. To address this, we developed the Prostate Cancer Genius App (PCGA), a smartphone-based intervention that provides PCa education and navigation for a user-friendly, CLIA-certified at-home PSA test. In a pilot RCT of 94 Black men aged 55–69, PCGA users demonstrated significantly greater PCa knowledge (p<0.05) and higher app usability (p<0.05) compared to the Prevention Taskforce App (PTFA), a general screening tool developed by the U.S. Preventive Services Task Force. PCGA users also showed higher PSA test activation (51.2% vs. 43.9%) and completion (83.3% vs. 50%), suggesting improved engagement and clearer navigation. This fully powered RCT will evaluate PCGA’s efficacy compared to PTFA and usual care (UC) among 684 Black men aged 55–69 in Oklahoma who are not up to date with PSA screening. Participants will be randomized 1:1:1 into PCGA (n=228), PTFA (n=228), or UC (n=228) and engage with their assigned intervention for 30 days, with follow-up for 6 months. The study will evaluate PSA screening completion (primary outcome) and screening readiness (i.e., knowledge, intentions, motivation, self-efficacy), diagnostic follow-up, treatment initiation, and satisfaction with care. Additionally, we will assess social needs as a moderator of screening and follow-up outcomes. Aligned with NOT-CA-24-033, this study will provide valuable insights into scalable, home-based PSA screening interventions that integrate telehealth-based shared decision-making. Findings will inform strategies to improve screening rates and follow-up care using tailored digital health tools. This research will also explore the role of clinic-based vs. home-based screening pathways to determine the most effective model for increasing PSA screening uptake. By rigorously assessing intervention engagement, screening readiness, and follow-up outcomes, this study aims to provide critical evidence for a sustainable and accessible approach to PCa early detection.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Candidate: Trained as a communication scientist with postdoctoral training in tobacco control, my career goal is to become a leading independent investigator in tobacco control by developing effective digital interventions to prevent and reduce tobacco use. Career development plan: I require essential training in three areas to complement my communication expertise, enabling me to implement the proposed study in the K01 and advance toward independence: 1) randomized clinical trial (RCT) design and implementation; 2) advanced biostatistical methods for clinical research; and 3) mHealth interventions for tobacco cessation. I have assembled a mentorship team of experts: Dr. Businelle (mHealth interventions for tobacco cessation), Dr. Ling (tobacco treatment on social media), Dr. Rigotti (clinical trials for tobacco treatment), and Dr. Frank-Pearce (advanced biostatistics). Environment: The University of Oklahoma Health Sciences Center (OUHSC) has a strong record of research on tobacco control and technology-based interventions, providing rich physical resources and an intellectually outstanding environment for the proposed training and research. Research Project: One commonly reported challenge that undermines the efficacy of digital interventions and impedes retention is significantly reduced participant engagement over time. Informed by theories and literature, the proposed study aims to address this issue by enhancing social support from peers in interventions. Building on my NCI-funded F32 study that developed a peer mentoring program (PEERS) to help young people quit vaping, the proposed study will test the feasibility, acceptability, and preliminary efficacy of a program named PEERS+QTH that incorporates PEERS into an existing social media-based vaping cessation intervention for young adults (QTH). It has three specific aims. Aim 1 is to test the feasibility and acceptability of the PEERS+QTH program by conducting a two-arm pilot RCT (PEERS+QTH vs. Smokefree.gov); Aim 2 is to gain qualitative insights from both the PEERS+QTH participants and peer mentors via post-intervention interviews to refine the program and study procedures for the next-phase RCT; Aim 3 is to assess the preliminary efficacy of PEERS+QTH by conducting a three-arm pilot RCT (PEERS+QTH vs. QTH vs. Smokefree.gov). Summary: The proposed study innovatively integrates peer mentoring with social media-based vaping interventions for young adults to enhance intervention efficacy and address a pressing health issue. It aligns with NIDA’s priority scientific area of "Develop and Test Novel Prevention, Treatment, Harm Reduction, and Recovery Support Strategies. My prior experience, along with the exceptional mentoring team and institutional environment, positions me well to pursue this K01. The training will fill the crucial gaps in my knowledge and skills, enabling me to successfully carry out the proposed research and establish a sustainable line of tobacco research. The K01 will also provide me with the necessary practical experience and preliminary data for a full-scale RCT in an R01 to rigorously test the efficacy of PEERS+QTH, facilitating my transition to independence.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Acute myeloid leukemia (AML) is a rare childhood malignancy that represents 15-20% of all leukemia diagnoses among children. Although survival rates among patients diagnosed with pediatric AML have improved over the past several decades (from <30% in the 1970s to approximately 70% today), overall survival still lags childhood leukemia overall (87%). Furthermore, contemporary curative therapy for pediatric AML is associated with profound acute and long-term toxicity, including an increased risk of cardiovascular events. In fact, nearly 50% of patients demonstrate evidence of cardiac dysfunction during treatment and an estimated 12% of survivors will develop premature heart failure. Our long-term goal is to improve outcomes among children and adolescents with AML through an improved understanding of the genetic etiology of adverse outcomes. While well- established prognostic factors for AML include both clinical and disease features, important gaps remain unanswered: 1) to what extent does inherited genetic variation contribute to variability in disease outcomes, and 2) does development of a polygenic risk score improve our ability to identify patients at risk of significant cardiotoxicity. This application pursues the central hypothesis that germline genetic variation in critical genetic pathways contributes significantly to individual susceptibility to unfavorable response to contemporary AML therapy. To evaluate this hypothesis, this study proposes two aims: 1) identify germline genetic variation in pharmacogenetics and cancer predisposition genes associated with AML overall and event-free survival, and 2) Develop and optimize a multi-ancestry polygenic risk score for treatment-associated cardiotoxicity in pediatric patients with AML. To accomplish these aims, this proposal leverages Gabriella Miller Kids First germline whole genome sequencing data generated on 1,126 AML patients and rich phenotype and outcomes data collected on these patients as part of the Children’s Oncology Group AAML1031 clinical trial. The investigative team is uniquely positioned to accomplish the proposed study. This application has the potential to advance our understanding of how germline genetic variation contributes to differences in AML survival and treatment- associated toxicity among children and adolescents. Ultimately, this work may guide clinical decision making regarding available treatment modalities, inform the delivery of targeted therapies, and assist with genetic counseling and screening strategies for patients and their families.

NIH Research Projects · FY 2026 · 2026-05

The COBRE program in this application is designed to develop the field of sensory biology on the OUHSC campus by supporting innovative research focusing on the processes used by cells to sense their environment. The COBRE will support a wide spectrum of projects using a variety of model organisms, from worms and green algae to mice and human tissue. It will support studies on cell signaling across the plasma membrane mediated by sensory organelles such as cilia and flagella; specialized signaling domains on the cell surface such as dendrites and immunological synapses; and cell surface molecules such as receptors and ion channels. It will also support research projects investigating disease and syndromes associated with sensory defects. Initially, the Oklahoma Center for Biomedical Research Excellence in Sensory Biology (OCBRESB) will take advantage of the rapidly expanding talent and expertise in cilia biology and diseases associated with defects in primary cilia on campus and then will expand to all sensory modalities. The essential biological and biomedical importance of cilia biology and the growing nexus of cilia biology researchers at OUHSC makes us uniquely positioned to develop a multidisciplinary biomedical Center of Excellence. OCBRESB will consist of an Administrative Core led by Dr. Leonidas Tsiokas, Professor and Chair of the Department of Cell Biology with research expertise on cilia biology and ion channel signaling, four Research Projects exploring mechanisms of sensory signaling in diseases affecting vision (Bennett), neurodevelopment (Craft Van de Weghe), neuronal function (Zhang), and neuropsychiatric disorders (Paterno), and two Research Cores, which will provide state-of the art support in super-resolution imaging and cell and genetic engineering. Strategically, the OCBRESB integrates perfectly well with OUHSC’s ongoing research on obesity/diabetes, vision, cancer biology; builds on the strength of Geroscience/Healthy Brain Aging; and functions as the springboard for new research activities in Neuroscience. Critically, it supports cutting-edge infrastructure needed for basic science research, traditionally the engine for discovery at OUHSC, to excel and achieve our short- and long- term strategic goals. Finally, it is essential for the development of the next generation of independent, NIH-funded talented investigators who will become leaders in an emerging area of highly relevant biomedical research. Completion of the OCBRESB will have a significant impact in enhancing biomedical research in the state of Oklahoma.

NIH Research Projects · FY 2026 · 2026-04

Affecting over 12 million people worldwide, Autosomal Dominant Polycystic Kidney Disease (ADPKD) is one of the most common inherited kidney diseases and is caused primarily by mutations in two genes—PKD1 and PKD2. In the 30 years since the identification of these genes, extensive effort has been made to understand how their protein products function under normal conditions and further, how naturally occurring mutations alter this function in ADPKD. While significant progress has been made to characterize PKD2 as a Ca2+-permeable nonselective ion channel, the function of PKD1 has proven more elusive. PKD1 has been proposed to act as an atypical G protein-coupled receptor (GPCR), but studies to define this function are limited by the absence of a known activating stimulus and lack of a typical 7-transmembrane (7-TM) domain signature present in all known GPCRs. We have discovered secreted WNT proteins as the activating ligands of the PKD1/PKD2 complex, defining this complex as a distinct class of WNT receptors. We asked the question of whether PKD1 functions as a WNT-activated GPCR. Using super-resolution live cell imaging and bioluminescence resonance energy transfer-based G protein biosensors, we find that PKD1 is directly coupled to specific subsets of heterotrimeric G proteins, such as Gαi1-3 and Gαq, but not Gαs or Gα12/13 in a WNT ligand-dependent manner. Specifically, we show that WNT ligands induce: 1) PKD1-mediated dissociation of Gα from Gβγ subunits, 2) time-dependent recruitment of Gα subunits to PKD1, and 3) Gα-GDP to -GTP transition via PKD1. The WNT-induced coupling of PKD1 to Gαi1-3 inhibits basal and forskolin-induced cAMP accumulation. PKD2 has an essential role in enabling PKD1 to function as a GPCR by chaperoning it to the plasma membrane. Naturally occurring mutations in PKD1 or PKD2 that compromise cell surface expression, assembly of the PKD1/PKD2 complex, or disrupt G protein recruitment diminish WNT-induced PKD1-mediated GPCR signaling. These data, which form the premise of this application, define the molecular function of the Polycystin complex and provide a direct mechanistic link of the complex to cAMP metabolism. We propose complementary in vitro and in vivo genetic approaches to take these findings to the next level to determine whether the direct coupling of PKD1 to Gαi and cAMP signaling is critical for the initiation of cystogenesis. Successful completion of the proposed studies will remove previously impossible technical and conceptual roadblocks to determine how naturally occurring mutations alter the function of these complex molecules and lead towards the development of new and more effective treatments for ADPKD.

NIH Research Projects · FY 2026 · 2026-03

ABSTRACT The brain consumes nearly 20% of the body’s ATP despite accounting for only 2% of body weight, highlighting its disproportionate energy demand. As the cells of the brain also exhibit a limited regenerative capacity, sustained bioenergetic failure often results in a permanent loss of cognitive, sensorimotor, and autonomic function. While neurodegenerative and neurodevelopmental conditions are often multifactorial, one common theme observed across these diseases is altered metabolic homeostasis. To meet the tremendous energetic demands of intercellular communication, neurons rely on highly efficient ATP production via mitochondrial oxidative phosphorylation (OxPhos). In addition to ATP, OxPhos yields reactive oxygen species (ROS) shown to oxidize cellular lipids, proteins, and nucleic acids. While decades of research have investigated the consequences of ROS accumulation in various pathologies and aging, the role of ROS in healthy neuronal function is incompletely understood. Membrane-diffusible mitochondrial hydrogen peroxide (mtH2O2), a specific form of ROS, has become the focus of recent research due to its reported interactions with redox modifiable cysteine residues – coined redox switches. As mtH2O2 is produced in the mitochondrial matrix in a metabolic performance dependent manner, it likely exhibits direct influence on local mitochondrial dynamics to shape metabolism and maintain a healthy mitochondrial network. Our preliminary data in primary mouse cortical neurons suggests that mtH2O2 serves a regulatory role in mitochondrial fission/fusion balance. Following attenuation of mtH2O2 via depletion of superoxide dismutase 2 (SOD2), we observed a reduction in mitochondrial size within neuronal dendrites. To ascertain if this change in size is a result of impaired mitochondrial fission/fusion balance, we performed time lapse fluorescent microscopy on live primary neurons and found that while SOD2 ablation decreases the frequency of both fission and fusion events, it preferentially inhibits fusion. Thus, we hypothesize that mtH2O2 dictates local neuronal mitochondrial dynamics. We have devised two independent aims to test this hypothesis. In the first aim, we will determine the compartment- specific signaling capacity of mtH2O2 by visualizing H2O2 spatial abundance in the mitochondrial matrix and at the outer mitochondrial membrane in neuronal dendrites, somas and axons using a novel genetically encoded H2O2 reporter, HyPer7. In the second aim, we will quantify changes in mitochondrial fission and fusion following genetic manipulation of mitochondrial antioxidants – resulting in physiologically relevant changes in mtH2O2 levels – then employ targeted proteomics to identify differences in the oxidation profiles of known fission and fusion proteins. If successful, our work will characterize an essential role of mtH2O2 in mitochondrial health by its direct regulation of proteins involved in mitochondrial remodeling.

NIH Research Projects · FY 2026 · 2026-01

PROJECT SUMMARY/ABSTRACT Chronic kidney disease (CKD) is an urgent health problem in the U.S., afflicting ~15% of Americans and costing the healthcare system $120 billion/year. Despite the availability of glucose-normalizing drugs, diabetes remains the leading cause of renal failure. Current treatments depend on glucose and hypertension control that cannot completely prevent diabetic nephropathy and progressive renal dysfunction. A major feature of diabetic nephropathy is inflammation, driven by the upregulation of adhesion proteins in renal endothelium and subsequent recruitment of inflammatory immune cells into renal tissue. Previous work has shown that knockout of the adhesion protein ICAM1 attenuates diabetic nephropathy in mouse models of type 1 and 2 diabetes. However, there is a lack of clinically viable technologies that can act on these findings. An FDA-approved lipid nanoparticle (LNP) siRNA drug, patisiran, is capable of safely inducing efficient (>80%) and durable (up to one month) gene knockdown in humans. As LNPs typically accumulate in the liver, this approach has not been broadly applicable to other tissues. This application seeks to develop new LNPs for efficient siRNA delivery to renal endothelium, to test the hypothesis that knockdown of ICAM1 is protective against type 1 and 2 diabetic nephropathy. The goal of Aim 1 (K99 phase) is to identify LNPs capable of efficient siRNA delivery to renal endothelium through high-throughput in vivo screening. A panel of DNA-barcoded LNPs with varying compositions will be screened for functional siRNA delivery to the renal endothelium using a new workflow. Isolated barcodes will be analyzed by deep sequencing to deduce LNP parameters that mediate renal endothelial delivery and inform subsequent, refined screens. Lead LNPs will be individually validated and assessed for knockdown efficiency, duration, and safety. During the R00 phase, LNP efficacy will be tested in multiple models of type 1 and 2 diabetic nephropathy. These models capture human diabetic nephropathy features of ICAM1 upregulation, albuminuria, renal fibrosis, and macrophage infiltration into renal tissue. LNPs carrying ICAM1 siRNA will be tested for efficacy in prophylactic (before disease onset) and therapeutic (after disease onset) models. Successful completion of this work could enable new, precision medicine approaches that target inflammatory drivers of diabetic nephropathy. The PI, Gary Liu, aims to lead a lab that develops new renal therapeutics and arrest disease progression. A major focus will be gene therapies for kidney diseases. To prepare him for this role, this K99/R00 application includes training and coursework in DNA barcoding, deep sequencing, and bioinformatics; nephrology and diabetic nephropathy; LNP technology; and mentoring practices for undergraduate, graduate, and postdoctoral trainees interested in medicine, academia, or industry. Moreover, this application will enable the PI to disseminate work at conferences, network, and apply for faculty positions. Training will occur at the Koch Institute of MIT in the laboratories of Profs. Robert Langer and Giovanni Traverso, which will provide the resources and intellectual expertise necessary to carry out the work.

NIH Research Projects · FY 2026 · 2025-12

PROJECT SUMMARY/ABSTRACT Despite decades of progress, coronary artery disease (CAD) remains the top global cause of death. While hundreds of CAD-associated genes have been identified, the translation into new treatments has been hampered by a lack of understanding of the molecular mechanisms tying these genes to disease risk. Although smooth muscle cells (SMCs) harbor much of the attributable risk for CAD and comprise the main cellular component of diseased vessels, no currently available treatments target these cells directly. As CAD progresses, SMCs undergo a complex process of phenotypic modulation, where SMCs migrate, proliferate, de- differentiate, and transition into other cellular states, including protective “fibromyocytes” (FMC). The candidate, Dr. Brian Palmisano, has developed strong preliminary data that support the CAD- associated gene, PR domain containing 16 (PRDM16), is a critical regulator of SMC phenotypic modulation. PRDM16 is an epigenetic modifier involved in cell state transitions in several cell types, and is highly expressed in vascular tissues, but its role in SMC phenotypic modulation and atherosclerosis is currently unknown. The candidate’s preliminary data indicate that SMC-specific loss of Prdm16 in an animal model of atherosclerosis leads to an increase in FMC development. In human coronary SMCs, overexpression of PRDM16 regulates SMC phenotypic modulation. As an epigenetic modifier with histone methyltransferase activity, PRDM16 also altered the enrichment of several histone methylation marks at genes related to SMC phenotypic modulation. This led to the candidate’s central hypothesis that PRDM16 regulates SMC phenotypic modulation and the transition to FMCs through its epigenetic modification of chromatin substrates, which will be tested in three specific aims. Aim 1 will determine the regulatory mechanisms by which PRDM16 suppresses SMC phenotypic modulation in vitro. Aim 2 will determine the gene regulatory networks underlying Prdm16-mediated phenotypic modulation of SMCs in vivo. Aim 3 will determine the downstream factors governing PRDM16-mediated phenotypic modulation of SMCs in vitro. The enclosed training plan encompasses research, mentoring and career development activities that have been specifically formulated to develop the scientific and professional skills necessary for Dr. Palmisano to transition to an independent physician scientist studying novel molecular mechanisms of atherosclerosis. Dr. Palmisano will develop his skills in bioinformatic integration of multi-omic datasets and smooth muscle cell function under the mentorship of his primary mentor, Dr. Tom Quertermous, co-mentor, Dr. Josh Knowles and his expert advisory committee, each with expertise in multi-omic data analysis and mentorship of physician scientists. Collectively, these studies will identify the molecular mechanisms by which the CAD-associated gene PRDM16 regulates SMC phenotypic modulation, a critical process related to CAD disease risk, allowing Dr. Palmisano to launch his independent career as a physician scientist.

NIH Research Projects · FY 2025 · 2025-09

NIH Research Projects · FY 2025 · 2025-09

Project Summary Epidemiological data show increased incidence and severity of asthma in women, which likely reflects both inherent sex differences and importantly the effects of sex steroids, especially estrogens. Our previous studies show upregulation of estrogen receptors (ERs), ERα and ERβ in human airway smooth muscle (ASM) with differential ERα vs. ERβ signaling in regulating ASM [Ca2+]i/remodeling and airway hyperresponsiveness (AHR). However, it is unclear whether it is estrogen that is responsible or its metabolites, locally produced in ASM. Data from other systems suggest differential physiological effects of cytochromeP450 (CYP’s) mediated estrogen metabolites, particularly 16αHE2 (via CYP3A4), promoting inflammation and remodeling. In this regard, some unresolved questions are A) What are the expression profiles and activities of ASM CYP’s and how do they influence local estrogen metabolism in asthma? B) What roles do specific estrogen metabolites play in regulating AHR and/or remodeling? Preliminary studies show: 1) Functional CYP’s are expressed in human ASM with higher CYP3A4 expression and activity in female asthmatics; 2) E2 with CYP3A4 inhibition showed reduced [Ca2+]i in asthmatic ASM; 3) Inhibition of CYP3A4 decreases E2 mediated effect on ASM proliferation and ECM deposition; and 4) In a mixed allergen (MA) mouse model of asthma, 16αHE2 exacerbates AHR and remodeling while 2-HE is alleviating. Thus, our overall hypothesis is that the differential role of estrogen- metabolizing enzymes (particularly CYP3A4 and its product 16αHE2) contributes to detrimental estrogen effects on airway contractility and remodeling in asthma. This will be tested via the following Specific Aims 1. To determine the expression and function of estrogen-metabolizing enzymes and their influence on [Ca2+]i and contractility in non-asthmatic vs. asthmatic human ASM. 2. To identify mechanisms by which estrogen metabolites influence remodeling in non-asthmatic vs. asthmatic human ASM. 3. To establish the effects of estrogen metabolites on AHR and remodeling in a mouse model of asthma. Using human ASM from non- asthmatic vs. asthmatic, females and males, Aim 1 explores the effect of inflammation and asthma on the expression and activity of CYPs relevant to functional estrogen metabolites and their effects on ASM [Ca2+]i/contractility. Aim 2 explores the role of estrogen metabolites in ASM proliferation, remodeling and downstream signaling intermediates. Aim 3 explores how estrogen metabolites affect airway structure (histology, laser capture microdissection) and function (flexiVent) in vivo. Endogenous hormone effects will be assessed using ovariectomy (OVX), and the role of ASM per se using smooth muscle-specific ERαfl/fl/smMHCCre/0 and ERβfl/fl/smMHCCre/0 knockout mice. Clinical significance lies in understanding the contribution of estrogen and its metabolites on ASM structure and function in chronic asthma in women. This will provide us with the basic fundamental knowledge to understand the greater prevalence of asthma in women and will help to develop a novel therapeutic approach targeting estrogen derivatives vs. estrogen signaling pathways.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Lipids constitute ~50% of the brain’s dry weight, yet very little is known about how brain lipid metabolism is regulated in both physiological and pathological states. Lipid droplets (LDs) are lipid-storing organelles that play a significant role in regulating cellular lipid metabolism. Dysregulated lipid homeostasis, resulting in excessive accumulation of LDs in brain cells, has been implicated in neuroinflammation associated with aging and other neurodegenerative diseases. While most studies have focused on lipotoxicity in glial cells, our single-cell RNA-sequencing (scRNA-seq) analysis reveals that brain endothelial cells are equally vulnerable to age-related LD accumulation. Endothelial cells accumulating LDs in the aged brain displayed a transcriptomic signature associated with reduced LD turnover, increased neuroinflammation, and impaired blood-brain barrier (BBB) function. However, the mechanisms by which age-related lipid dysregulation in brain vascular cells, specifically endothelial cells, contribute to cognitive impairment remain poorly understood. In this novel pilot study, we build on our exciting preliminary data and propose that decreased lipid turnover induces microvascular endothelial dysfunction, which impairs cerebral blood flow responses, promotes BBB disruption, and triggers neuroinflammation, ultimately leading to cognitive decline in aging. To test this hypothesis, we will knock down the critical triglyceride hydrolytic enzyme, adipose triglyceride lipase (ATGL), specifically in brain endothelial cells using the recently developed AAV serotype BI30. We will utilize this model to evaluate how LD accumulation in endothelial cells drives microvascular aging and cognitive impairment. In Aim 1, we will assess the effects of LD accumulation on endothelial structure and function. In Aim 2, we will evaluate how endothelial LD accumulation impacts endothelial bioenergetics and cognitive function in aging. The successful completion of these studies will pave the way for novel therapeutic strategies targeting endothelial lipid metabolism to delay or prevent cognitive decline associated with aging and neurodegenerative diseases.

NIH Research Projects · FY 2025 · 2025-09

Acute kidney injury (AKI) is defined as an abrupt, and often reversible, decline in kidney function that is characterized by a decrease of urine output, an increase in creatinine, or both. The ischemia reperfusion injury (IRI) model most effectively recapitulates the pathophysiological conditions observed following AKI. Mechanistically, post-ischemic tubular epithelial cells (TECs) undergo necrosis and secrete damage associated molecular patterns (DAMPs), initiating proinflammatory responses by recruiting monocytes, neutrophils, dendritic cells and other immune cells. Prolonged inflammation results in further damage to tubular cells, and eventually CKD. Data from our lab and others has identified macrophages as a key contributor to the injury and repair process after IRI; however, a detailed description of macrophage phenotype at a cell-by-cell level after IRI has not been performed and represents a significant gap in knowledge. To address this gap in knowledge, we generated a comprehensive single-cell RNA (scRNA) sequencing dataset of macrophages at early, intermediate, and late stages post IRI. Using this dataset, we identified two injury-specific macrophage clusters: one predominant during the early inflammatory phases (injury-specific macrophage 1 or ISM1), and the other emerging during the reparative phase of IRI (injury-specific macrophage 2 or ISM2). A detailed analysis revealed that ISM2 highly expressed Triggering receptor expressed on myeloid cells 2 (Trem2), a phagocytic receptor involved in anti-inflammatory signaling. Temporal analysis revealed that Trem2 expression begins 2 days post injury and persists until day 28. CellChat analysis predicted that ISM2 promote tubular repair after injury by producing reparative ligands including IGF1 that bind to cognate receptors on injured TECs. Igf1 has previously been associated with improved kidney function after injury by activating cell survival and cell cycle in an ERK/MAPK and Akt/Rb dependent manner. Based on these data, I hypothesize that the TREM2 receptor is required for transition of macrophages from a pro-inflammatory to pro-reparative state, and that ISM2 derived Igf1 promotes tubular epithelial cell repair after injury. In this application, I will test these hypotheses through a mixture of in vivo and in vitro techniques. There are currently no FDA approved therapies to treat AKI, or to prevent the AKI to CKD transition. Treatments for patients developing CKD are palliative in nature and limited to dialysis and transplant, both of which pose a significant health care burden. Thus, there is an urgent need to develop new therapies to treat AKI and CKD. An understanding at the molecular level of the AKI-to-CKD transition will aid in the development of effective treatments. Further, the comprehensive training plan will provide me with the skills necessary to become an independent investigator study the function of macrophages after AKI.

NIH Research Projects · FY 2025 · 2025-09

Abstract/Summary Lung cancer mortality in Oklahoma is higher than the national average with age adjusted rate of 46.5 compared to 35 per 100,000 population. I am a thoracic medical oncologist with high interest in clinical and translational research. I joined Stephenson Cancer Center of the University of Oklahoma Health Sciences Center in 2014 with the goal of improving lung cancer care in the state. In 2015, I joined the lung committee of SWOG Cancer Research Network, and I have been an active member of that group since then. I regularly participate in monthly virtual meetings and bi-annual in-person meetings of that group to explore and evaluate new ideas to improve care of lung cancer patients. I worked with my colleagues in that committee to develop a study to improve the care of stage 2 and 3 non-small cell lung cancer (NSCLC) patients with borderline performance status. That study, SWOG 1933, is currently open to enrollment and I am the national PI on it. I also, proposed a research concept to study perioperative versus adjuvant chemo-immunotherapy in early-stage NSCLC and we submitted that study through SWOG. I worked with my colleagues in the Alliance Cancer Research Network, and we jointly received the first Clinical Trial Innovation Unit (CTIU) grant to run that study as CITU 2317. That study would likely define the new standard of care in early-stage NSCLC. I joined the NRG Oncology Research Network in 2019, and I have been an active member of the lung core committee of that group since then and I regularly attend their meetings. I am also the medical director of the clinical trial office in our institution and a local PI on phase one and later phase thoracic oncology studies. The R50 Research Specialist Award will allow me to continue to be an active member of the NCI cooperative groups, to develop innovative clinical trials to improve lung cancer care in Oklahoma and mentor junior faculty and fellows in our institution in the field of clinical and translational research.

NIH Research Projects · FY 2025 · 2025-09

Abstract Systemic lupus erythematosus (SLE) is a heterogeneous disease affecting multiple organ systems. Current treatments result in toxicity and impaired immune responses to pathogens. Identification of new therapeutic targets is complicated by a lack of understanding of the underlying mechanisms that result in SLE and in other autoimmune diseases. We found that ARID3a (A+T-rich interaction domain protein 3a) over-expression in transgenic mouse B lymphocytes was sufficient to cause autoantibody production and immune complex formation in kidney glomeruli. Further, numbers of ARID3a+ B cells are increased in SLE patients compared to healthy controls and those numbers are highly associated with increased disease activity in patients as determined by disease activity indices (SLEDAI scores). Although ARID3a is a DNA-binding protein involved in gene regulation, mechanisms by which ARID3a contributes to gene dysregulation in autoimmunity are unknown. Naïve B cells represent a tolerance check-point and we observed that a subset of naïve B cells from SLE patients express ARID3a, while ARID3a was absent from healthy control naïve B cells. We recently used single cell RNA-seq to identify genes differentially expressed in naïve ARID3a+ versus ARID3a- B cells from SLE patients, identifying new markers to enable additional functional studies of those cells. We also found that complex stimuli developed by others to generate “activated” naïve B cells and pathogenic double--negative B cell subsets similar to those increased in SLE patients, also induced ARID3a expression in the “activated” naïve B cell subset. Therefore, we hypothesize that ARID3a is a mediator of dysregulated gene expression in SLE naïve B cells that contributes to tolerance breaches. The proposed aims will 1) define genes and epigenetic regions that bind ARID3a and may be directly regulated by ARID3a expression; 2) determine how ARID3a contributes to functions associated with breaches in B cell tolerance; and 3) determine if inhibition of ARID3a-regulated pathways in autoimmune mouse models ameliorates disease symptoms and functional attributes. The proposed studies will fill gaps in our knowledge of how ARID3a functions in naïve B cells and will define genes and intragenic regions regulated by ARID3a that may contribute to autoimmune characteristics in SLE. These experiments will provide new mechanistic insights into ARID3a functions and may direct future design of novel therapeutics.

NIH Research Projects · FY 2025 · 2025-09

I am the Stevenson Chair and Associate Professor of Urologic Oncology, as well as Adjunct Associate Professor of Medical Oncology, at the Stephenson Cancer Center (SCC), University of Oklahoma College of Medicine, where I am deeply involved in advancing NCI-sponsored clinical research. I hold leadership roles within the National Clinical Trials Network (NCTN) as a SWOG Genitourinary (GU) Committee Board Member, GU Liaison to the Surgery Committee, and Site PI for SCC GU clinical trials. At the Oklahoma City Veterans Administration Medical Center (OKC-VAMC), I am the SWOG Site PI and have established a program offering NCI-sponsored trials to veterans who previously had limited options. As both a surgeon and clinical trialist, I have developed investigator-initiated trials that uniquely integrate surgical interventions with emerging therapeutics. Additionally, I mentor junior investigators in clinical trial development, supporting them as local PIs of NCTN trials and investigator-initiated trials (IITs). At SCC, I chair the Data and Safety Monitoring Committee (DSMC), overseeing clinical trial data integrity and patient safety, and serve as Disease Site Chair for GU oncology, where I lead a multidisciplinary team and manage SCC’s clinical trial portfolio. Under my leadership, SCC has maintained top-tier enrollment rates among NCTN Lead Academic Participating Sites (LAPS). My work is essential to SCC’s growth as an NCI-designated Cancer Center. Through this application, I aim to 1) leverage novel mHealth applications to boost NCI trial accrual, 2) promote clinical trial enrollment representative of our catchment area population, 3) expand veteran access to NCI-sponsored trials, and 4) create leadership opportunities for SCC investigators in NCI-sponsored trials. An R50 Clinical Research Specialist award would allow SCC to protect my time for these impactful research activities. Achieving these goals will significantly enhance SCC's clinical research enterprise and contribute meaningfully to the NCI’s mission.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT Bacterial eye infections cause a significant number of cases of blindness worldwide. Bacteria secrete an armamentarium of virulence factors that protect them from killing by immune cells. Most pathogens that infect the eye produce a polysaccharide capsule that covers the bacterial surface. This “cloaking” renders them invisible to receptors and pathways that would otherwise incite an immune response, protecting them from killing by immune cells. Unhindered growth results in virulence factor production. In the eye, this can manifest as an irreversibly damaged retina and significant vision loss. To make matters worse, many of these pathogens belong to the ESKAPE group (Enterococcus, Staphylococcus, Klebsiella, Acinetobacter, Pseudomonas, Enterobacter) and are at the top of the CDC’s list of Antibiotic Resistance Threats in the US. Multidrug resistance is common in ocular isolates, elevating the risk of infections that are difficult to treat. Our program investigates the pathogenic mechanisms underlying ocular bacterial infections. For bacteria, animal infection models and ocular cell lines are used to investigate the areas noted above. We have a very good idea of the virulence factors that directly damage the retina and the innate pathways involved in inflammation. This proposal is focused on a common virulence factor which may be involved in cloaking bacteria during the earliest stages of endophthalmitis, events that, to date, have drawn minimal attention. This new R01 proposal is based on the overarching hypothesis that ocular pathogens are cloaked from the intraocular immune response by capsule expression in the eye during infection. The scientific premise is based on data demonstrating that: 1) capsule genes are expresssed in intraocular environments and in infected eyes of mice, 2) capsule-deficient mutants are not as virulent as bacteria that express capsule, 3) capsules are potential PAMPs and may interact with TLRs, and 4) anti-capsule antibody improves the clinical outcome of endophthalmitis caused by streptococci and non-ocular infections caused by encapsulated pathogens. These gaps are investigated in three separate but related aims focused on capsule-mediated cloaking of ocular pathogens in endophthalmitis. We will use well-characterized capsule-deficient mutants in in vitro and in vivo models in rigorous and straightforward experiments designed to define the role of capsule in endophthalmitis and the potential value of therapeutically targeting capsule to improve clinical outcome. For patients with eye infections, ineffective treatment often equates with vision loss. Our approach to addressing these gaps in our field are innovative, translationally relevant, and will move the ocular infectious disease field forward by focusing on a common bacterial virulence factor that is targeted in FDA-approved vaccines which have improved the health of millions. These studies are a logical extension of our ocular infection research program, and we are well positioned to contribute new and important information that will improve options for preventing infection and preserving vision.

NIH Research Projects · FY 2025 · 2025-09

Vascular inflammation underlines common pathophysiology of various cardiovascular diseases and determines the progression of infections and autoimmune diseases. Several tetraspanin proteins are highly expressed in endothelial cells (ECs). Some of these tetraspanns inhibit vascular inflammatory responses including i) endothelial leakage, ii) endothelial recruitment of leukocytes, and iii) endothelial release of cytokines. Hence, we call them anti-inflammatory tetraspanins. It is unclear how anti-inflammatory tetraspanins inhibit these vascular inflammatory responses. The goal of this study is to identify the mechanisms by which anti-inflammatory tetraspanins restrain vascular inflammation at the molecular, cellular, and organism levels. We hypothesize that anti-inflammatory tetraspanins restrain the endosome-mediated i) presentation of inflammation- related cell adhesion proteins on the EC surface and ii) release of pro-inflammatory cytokines from ECs, to prevent and inhibit vascular inflammation. In this project, we will first assess the inhibitory roles of anti-inflammatory tetraspanins in inflammation-induced i) leakage and ii) damage of endothelia. Secondly, we will assess how anti- inflammatory tetraspanins prevent endothelial recruitments of leukocytes and platelets in inflammation by revealing anti-inflammatory tetraspanin effects on the i) levels, ii) activities, and iii) trafficking of inflammation-related cell adhesion proteins in ECs in inflammation. Finally, we will determine how anti-inflammatory tetraspanins regulate EC secretion of cytokines to prevent inflammation exacerbation by i) assessing in vivo and in vitro effects of anti-inflammatory tetraspanins on cytokine secretomes, ii) revealing anti-inflammatory tetraspanin roles in Weibel- Palade body formation and maturation, and iii) identifying the molecular machinery crucial for anti- inflammatory tetraspanin-regulated exocytosis. This project will fill important knowledge gaps regarding the molecular and cellular mechanisms that regulate vascular inflammation in cardiovascular and other diseases. This project will also establish tetraspanins as novel therapeutic candidates against vascular inflammation.

NIH Research Projects · FY 2025 · 2025-09

SUMMARY/ABSTRACT: Three receptor activity-modifying proteins (RAMP1-3) in humans form heterodimers with the class B G protein-coupled receptors (GPCRs) calcitonin receptor (CTR) and calcitonin-like receptor (CLR) and modulate their responses to six CGRP family peptide hormones. This complex system of seven receptors and six ligands is a model for understanding class B GPCR signaling mechanisms and modulation of GPCR function by accessory membrane proteins. CGRP family peptide signaling through CTR, CTR- RAMP1/2/3, or CLR-RAMP1/2/3, is involved in diverse processes including calcium homeostasis, blood glucose regulation, pain signaling, neuroimmune communication, and development and maintenance of the cardiovascular and lymphatic systems. Consequently, the receptors are drug targets for osteoporosis, diabetes, obesity, migraine headache, and cardiovascular and lymphatic disorders. We have used biochemistry, pharmacology, and structural biology approaches to study how the peptides bind and activate the receptors, how the RAMPs determine ligand selectivity, and how the peptide-bound receptors couple to G protein transducer to promote cAMP signaling. Our studies advanced our understanding of each of these areas and enabled us to engineer novel peptide antagonists and agonists with useful properties like increased receptor affinity, altered receptor selectivity, or long-acting signaling. These have been powerful tools for studying the receptors and may have promise as therapeutic leads. More recently, we discovered temporal differences in the signaling properties of the CGRP family peptides that likely contribute to their diverse biology. Some of the peptides promote cAMP signaling of short duration, whereas others elicit long duration signaling. Despite these advances, several areas need more study. How RAMPs determine ligand selectivity is only partially understood. Our understanding of the kinetics of peptide binding and cAMP signaling is incomplete, and the molecular basis of long duration signaling is unclear. Our understanding of the interactions of the receptors with the two other families of transducer proteins that mediate GPCR signaling, the GPCR kinases (GRKs) and b-arrestins, is rudimentary. Moreover, there is still a need for better pharmacological tools for studying the receptors. Accordingly, our goals are to further define the mechanisms by which RAMPs determine CTR and CLR ligand selectivity, further characterize the kinetics of the molecular events leading to cAMP signaling, and define the basis for long duration cAMP signaling. In addition, we will continue to engineer novel peptide ligands and we will study the receptor interactions with GRK and b-arrestin transducers.

NIH Research Projects · FY 2025 · 2025-08

Escalating geopolitical instability, nuclear proliferation, and vulnerability of medical, radiological and nuclear power infrastructure are real and constant threats raising the likelihood of catastrophic radiation exposure. Hematopoietic stem cells (HSCs) and hematopoiesis are among the most sensitive tissues/organs to radiation injury with multiple organ systems also affected. Since incidents are likely to be unanticipated, therapeutic strategies need to be effective even if commenced hours or days post-exposure, and administration routes should be compatible with treatment in the field. Currently, there are very few - if any - agents that can be used to rescue injury from lethal radiation doses after 24 hours. This gap represents a critical unmet need for both public health and the armed forces. The human stress-protectant protein thioredoxin (TRX) has shown exciting potential as a broad-acting radiation protective agent but does not have suitable pharmacological properties for use as a drug. Preliminary rodent and non-human primate (NHP) studies by the applicant with ORP100S, an engineered pharmacologically optimized version of TRX, have demonstrated remarkable radiation survival when ORP100S was administered subcutaneously 24 hours after irradiation. Long-term goal: develop ORP100S into a “deliverable” agent for treatment of lethal radiation-related injury. Overall objectives: further define the efficacy of ORP100S in mitigating radiation-induced injury and determine the molecular mechanisms through which ORP100S regulates and protects HSCs from radiation injury. Central hypothesis: ORP100S has broad, pan-cytoprotective effects on multi-organ systems and can mitigate lethal dose radiation. These hypotheses have been formulated based on data produced in the applicant’s laboratory. The rationale for the proposed research is that, once it is known how ORP100S protects HSCs and its effectiveness is confirmed in both mice and non-human primates, we will be able to move ORP100S forward into clinical use as a new and innovative therapeutic for radiation-related injury. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: Aim 1: determine the dose modifying factor of ORP100S and ability to rescue hPSC lung organoids from radiation injury. Efficacy of ORP100S in mitigating lethal radiation will be compared with the FDA approved agent (G-CSF) and any synergistic effects will be identified. Aim 2: define the molecular mechanisms through which ORP100S protects HSCs from radiation injury. Effects of ORP100S on the p53-ferroptosis pathway will be defined. Aim 3: determine efficacy, safety, PK, PD, and cytokine profiling of ORP100S in mitigating radiation-induced injury in non-human primates. The approach is innovative, because it focuses on a novel protein that is effective in mitigating toxic effects of radiation when given after 24 hr of radiation exposure. Humanized mouse model and hPSC induced lung organoids will be used and novel molecular pathway will be investigated. The proposed research is significant, because it will bring a new therapeutic agent to the national stockpile for the treatment of radiation injury.

NIH Research Projects · FY 2025 · 2025-08

SUMMARY Fungi can adapt and grow in a wide range of quickly changing environments, especially under harsh conditions inside a host. However, the fundamental mechanisms underlying fungal fitness and survival inside a host remain poorly understood. Stress responses during host colonization are relevant to human health, since pathogenic fungi with activated stress-responses are better colonizers and less susceptible to antimicrobial treatments. This results in more than 1 million deaths associated with fungal diseases each year globally. Along with antifungal compounds, these species have also evolved and adapted to elevated temperature (37oC), low O2 level, elevated CO2 level, and other stresses derived from the host immune system. This phenomenon raises several fundamental questions. How do fungi adapt and grow within a host’s environment during colonization? What are effects of the host immune system on fungal fitness, and how do fungi respond against these stimuli? Finally, how do shifts in growth environment, and negative effects of the immune system and antifungal compounds, have an impact on fungal plasma membrane biology? The proposed research program tackles these questions by taking advantage of the fungus Nakaseomyces glabratus as a tractable model organism. With the uses of fungal genetics, molecular biology techniques, next-generation sequencing, proteomics, lipidomics, and an in vivo mouse model, these studies will yield a new and more mechanistic understanding of how fungal cells sense and adapt to environmental stress during host colonization. The results will have broad implications for understanding the basic principle of stress response across many biological systems. They will also bring forth foundational knowledge that will be useful for human health improvement, in terms of future antifungal development and diagnostic strategies targeting environmental stress sensing and response.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Vascular occlusive disease remains a critical cardiovascular health issue and results in about 500,000 percutaneous-coronary angioplasties annually in the US. Restenosis due to neointimal hyperplasia occurs after 10-30% of angioplasties and remains a critical problem in cardiovascular medicine. No new approaches to improve the standard treatment, the insertion of mTOR inhibitor-or taxol-eluting stents, have been developed in a decade. Addressing this critical problem will require novel strategies to inhibit proliferation of vascular smooth muscle cells (VSMCs), the main driver of neointimal hyperplasia. Our long-term goal is to determine how mitochondrial function modulates cytosolic signaling events in VSMCs and endothelial cells in vascular disease. The overall objective of the proposed research is to determine how the highly conserved GTPase MIRO1, which resides in the outer mitochondrial membrane, regulates VSMC proliferation and neointima formation and test new mechanistic therapies. MIRO1 is known to control intracellular trafficking of mitochondria. Nascent data suggest that MIRO1 may have additional functions beyond mitochondrial trafficking. MIRO1 associates with the mitochondria-ER contact sites (MERCS) that facilitate mitochondrial Ca2+ entry and maintains the organization of mitochondrial cristae that is pivotal for electron transport chain (ETC) activity. Yet, the functional consequences of MIRO1 binding are not fully understood, and the implications for human disease, in particular in vascular disease, have remained unknown. We previously reported that mitochondrial Ca2+ uptake and energy production are required for cell- cycle progression in VSMCs in G1/S transition and beyond. Our pilot studies reveal that proliferation in VSMCs with MIRO1 deletion is abolished starting at G1/S transition. Thus, we speculate that MIRO1 controls VSMC proliferation via two hitherto unrecognized processes. Thus, our central hypothesis is that MIRO1 is required for VSMC proliferation, and thus for neointimal hyperplasia, by controlling the mitochondrial-ER Ca2+ transit and ETC activity. This is supported by our strong pilot data demonstrating that MIRO1 deletion blocks neointimal hyperplasia, VSMC proliferation and ATP production. Our novel tools and assays put us in a perfect position to test our hypothesis, including VSMC lines that lack MIRO1 or express MIRO1 mutants, and innovative RNA-aptamer-based tools for targeting of VSMCs in vivo after vascular injury. Our specific aims are 1. to determine the extent to which MIRO1 regulation of Ca2+ transport at MERCS controls cell-cycle progression, 2. to establish the extent to which the control of cristae organization by MIRO1 regulates ETC activity and proliferation, and 3. to test whether VSMC-specific aptamers that deliver siMIRO1 effectively reduce neointima hyperplasia. The anticipated outcomes of the proposed study are knowledge of the mechanisms by which MIRO1 promotes VSMC proliferation, and of the potential of RNA-based aptamers with siMIRO1 activity in preventing restenosis after vascular injury.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY/ABSTRACT This proposal is responsive to NIDA strategic plan initiatives focused on applying addiction science to improve public health and policy. In April 2022, the Food and Drug Administration (FDA) announced proposed rulemaking to ban all characterizing flavors other than tobacco (like Chery, Chocolate) from cigars.1 Cigarillos, the most widely sold cigar product, are predominantly used by young adults (YAs, defined here as ages 18-24), as well as Black and Hispanic individuals,2-5 and flavors are cited as a primary reason for their use.6-8 The emergence of “concept flavors” – vague ambiguous flavor descriptors with no explicit flavor name – is a product characteristic used by tobacco companies to evade local flavor bans9 and have been shown to appeal to YAs.10-12 Notably, of all other tobacco products, cigarillos show the greatest increase in concept flavor sales in recent years, compared to sales of tobacco and other characterizing flavors.6 Existing research, including our own, shows that explicit characterizing flavor descriptors on cigar products are associated with greater perceptions of product appeal (e.g., subjective effects like taste, smell, and enjoyment to smoke)13, 14 and use behavior14-16 relative to products labeled with tobacco flavor descriptors. Despite the rapid increase in sales of concept flavored cigarillos6 and early evidence of the appeal of e-cigarettes with concept descriptors17, the addiction potential of concept descriptors (e.g., subjective reinforcing effects, use behavior, motivation to consume/purchase), especially among younger cigarillo users, remains unknown. These indices are important indicators of the neurobiological systems that underlie addiction and motivate subsequent tobacco use.18, 19 Limited research exists, but this is all done in adults, or via online experiments or cross-sectional designs20-23, indicating the need for more studies. Only one laboratory study has investigated the impact of flavor on different indices of cigarillo use and appeal,20 but it did not include concept flavors and recruited only cigar-naïve cigarette smokers. There is a critical need to isolate the unique effects of concept descriptors on increased cigarillo use and appeal among experienced YA cigarillo users. Without this information, regulatory actions will only target characterizing, but not concept flavors, unintentionally shifting the cigar market to concept-flavored cigarillos. This would undermine the intended public health benefits of a cigar flavor ban. To address this need, this multi-method study will determine the addiction potential of concept flavored cigarillos in YA cigarillo users (N=300) by examining associations of cigarillo flavor type (concept, characterizing, tobacco) and their flavoring additives to complementary measures of product appeal: subjective effects (e.g., satisfaction, reward, taste), actual smoking behavior (e.g., number of puffs), and cigarillo purchasing, product substitution, or quitting under different flavor ban scenarios via an Experimental Tobacco Marketplace (ETM).15, 24, 25 This study will contribute to the evidence-base that informs the implementation of the cigar flavor ban, can fortify FDA’s claims in potential legal challenges to the ban, and inform the addiction potential of tobacco products with concept flavor descriptors.