West Virginia University

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Gadolinium-based contrast agents (GBCA) have revolutionized MRI. They provide contrast and essential diagnostic information that could not be obtained otherwise. For about 20 years, these compounds were considered among the safest pharmaceuticals. However, in 2006 the causal relationship between the development of a devastating and potentially fatal condition named nephrogenic systemic fibrosis (NSF) and GBCAs in renocompromized patients was established. The FDA has since placed restrictions on the use of GBCAs for patients with impaired kidney function, virtually eliminating the risk of NSF; however leaving some patients without having access to potentially lifesaving contrast-enhanced MRI. The more recent reports that gadolinium deposits cumulatively in the brain, bones, and skin, even in patients with intact blood-brain-barrier and normal renal and hepatobiliary functions, with unknown long-term hazard significance, have raised many concerns among the scientific community and patients. The last few years have seen breakthroughs in the development of metal-free MRI contrast agents based on macromolecular templates loaded with sterically shielded nitroxide radicals. These metal-free MRI contrast agents have demonstrated high efficacy in vivo with the ability to provide contrast in disease tissues (e.g., tumors). However, the current designs are limited by the instability of nitroxides radicals in vivo, leading to reduced diamagnetic hydroxylamines. The PI's lab has recently reported the synthesis of a highly biocompatible triarylmethyl radical with unmatched in vivo stability, named OX063. This radical has the capability to solve the stability issues of nitroxide-based organic radical contrast agents (ORCA). Moreover, Ox063’s trivalent shape makes it a natural dendrimer building block. In this project, we propose to develop OX063-based dendrimers as the next generation of metal-free MRI contrast agents. In specific aim 1, we will synthesize and characterize OX063-based dendrimers as dual MRI and fluorescent agents. A set of ORCAs will be decorated with a tumor-associated macrophage-binding peptide for active tumor targeting. In specific aim 2, we will perform in vitro and in vivo distribution, metabolism, excretion, toxicology, and pharmacokinetic studies, and in vivo imaging validation. Finally, in specific aim 3, we will demonstrate the application of OX063-based dendrimers for in vivo MRI tumor imaging in a mouse model of breast cancer. Active and passive targeting of the new ORCAs will be compared. The completion of the project will provide highly biostable metal-free contrast agents which can significantly impact the field of biomedical imaging.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY Ischemia-reperfusion (IR) injury is a significant challenge in treating myocardial infarction (MI), the leading cause of death in the United States. Mitochondrial reactive oxygen species (mtROS) generated by electron transport chain (ETC) Complex-I are the principal mediators of IR injury. Excess mtROS generated during early IR triggers vicious cycles of free radical production promoting cardiomyocyte death. Therefore, understanding the early molecular events of reperfusion will provide new targets for developing novel interventions for limiting cardiac injury. Our published findings show that LonP1- a major mitochondrial stress response protease mitigates oxidative stress-induced damage during early IR; therefore, LonP1 could be a promising target for attenuating reperfusion injury. Our long-term goal is to leverage the mitochondrial protein quality control mechanisms of LonP1 as a pivotal point to develop therapeutic strategies for mitigating IR injury and post MI- heart failure. Our published findings show that increased LonP1 expression in the heart induced by ischemic preconditioning (IPC) or transgenic overexpression (LonTg) reduced IR injury and favors cardioprotection. Whereas, LonP1 downregulation (LONP1+/-) abrogated IPC-mediated cardioprotection. Importantly, LonTg hearts showed reduced levels of Complex-I subunits (but not Complex II-V subunit) and oxidative damage during early IR (within 30 min reperfusion) compared to NTg controls. Conversely, our additional findings show that LonP1 downregulation in cardiomyocytes upregulated Complex-I activity, increased superoxide levels, and showed early reperfusion-induced cell death activation. In addition, we have identified a small molecule activator of LonP1 that significantly reduced hypoxia-reoxygenation (H/R) induced myocyte death in a dose-dependent manner in vitro. With additional data on IR-induced acetylation of Complex-I matrix subunits and LonP1 dependent Complex-I remodeling during IR, we hypothesize that LonP1 mitigates myocardial injury by suppressing excess mtROS generation through tight regulation of Complex-I during early IR. We will test our hypothesis by the following specific aims: Aim 1 will delineate the mechanism(s) by which LonP1 modulates Complex-I levels, activity and reduces oxidative stress during IR. Aim 2 will test that LonP1 remodels Complex-I and its associated supercomplexes by degrading IR-induced post-translationally modified (PTM) Complex-I matrix subunits, thereby reduce mtROS during early IR. Aim 3 will determine the therapeutic potential of LonP1 activators in treating myocardial IR injury in vivo. By determining the molecular mechanisms of LonP1-mediated cardioprotection and the therapeutic potential of LonP1 activators, we will define the role of LonP1 in cardioprotection and develop novel therapeutic tools and strategies to mitigate IR injury.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY/ABSTRACT Clinically, nearly all breast cancer patients at the time of diagnosis and prior to treatment have some degree of muscle dysfunction resulting in fatigue that ranges from mild to debilitating and may worsen during and after chemotherapy, radiation, and/or surgery. Adverse systemic effects of tumor growth can often result in treatment cessation and greater mortality in late stages of disease. The long-term goal of my work is to identify potential therapeutic targets for fatigue and a mechanism linking BC with systemic muscle fatigue. The specific goal of this proposal is to utilize our murine model to characterize the molecular adaptations in muscle and identify targets to attenuate fatigue in patients with breast cancer. The central research hypothesis is that regulation of mitochondrial bioenergetics via a PPARγ-agonist will attenuate breast tumor-associated muscle fatigue. Three Specific Aims have been proposed to test this hypothesis, using murine models of breast cancer and novel in vitro models of PPAR-activity. In Specific Aim 1, we will test the working hypothesis that breast tumor growth impairs mitochondrial bioenergetics resulting in ATP deficiency and subsequent muscle fatigue through aberrant function of mitochondrial electron transport chain (ETC) complex V. In Specific Aim 2, we will test the working hypothesis that breast tumor-derived miR-27a-3p interacts with regulatory components of PPARγ within skeletal muscle to decrease its function as a transcription factor, thereby specifically inducing alterations in mitochondrial function. In Specific Aim 3, will test the working hypothesis that pioglitazone will attenuate breast tumor-associated fatigue by upregulating PPARγ transcriptional activity in skeletal muscle, thereby rescuing mitochondrial bioenergetics and ATP production. BC-PDOX mice and controls treated with and without pioglitazone will be evaluated for muscle fatigue, mitochondrial bioenergetics and ATP content. This project is conceptually innovative in its use of a preclinical mouse model that phenotypically and transcriptionally mimics BC-associated muscle fatigue in the absence of cachexia. Our approach is both unique and practical in that it seeks to lay the foundation for repurposing an existing FDA-approved PPARγ-agonist for treatment of fatigue in patients with BC, directly addressing a key knowledge gap in this field. The outcomes of this project will impact the treatment of cancer- related fatigue, with the potential to offer early-stage BC-patients a treatment strategy targeting this debilitating symptom before the onset of cachexia. The aims and objectives of this project reflect the goals of the NCI, as described in their mission statement, by specifically conducting research that will advance scientific knowledge and be applicable to a large population of patients as well as helping improve patients’ quality of life during and following completion of cancer-associated therapy.

NIH Research Projects · FY 2025 · 2022-05

ABSTRACT Women have more rapid cardiovascular disease (CVD) risk development compared to men during young adulthood; yet, little research has studied factors that could curtail CVD risk development during this critical period for young women. The Nulliparous Pregnancy Outcomes Study Monitoring Mothers-to-Be Heart Health Study (nuMoM2b HHS) is a unique, multi-center, longitudinal cohort, originating during the first pregnancy, that is now studying associations between adverse pregnancy outcomes (APO) and modifiable CVD risk in women. This ancillary application to nuMoM2b HHS aims to test our hypotheses that sedentary behavior (SED) is a key, modifiable risk factor for CVD risk development in young women, including those with APO. SED is low-intensity behavior in a seated, reclining, or lying posture and has recently been identified as a CVD risk factor that is distinct from insufficient moderate-to-vigorous intensity activity (MVPA). Acute prolonged sitting results in marked, adverse responses such as increased blood pressure (BP), glucose, and lipids. Yet, major research gaps preclude the development and testing of specific SED-reduction interventions, especially for young women. Most studies measure SED poorly (e.g., by self-report) and do not statistically consider that SED is part of an all-day activity pattern (or composition). Further, mechanisms of how SED leads to CVD are unclear, further limiting intervention development. Last, almost no studies focus on young women. Our preliminary data suggest that SED is strongly associated with APOs and reduced cardiovascular health (e.g., BP) in young women post-pregnancy. Further, our laboratory has recently demonstrated novel associations between SED and mechanistic CVD measures, including greater arterial stiffness (higher pulse wave velocity [PWV]) and worse autonomic function (lower heart rate variability [HRV]). Collectively, SED is a novel, understudied risk factor for reduced cardiovascular health that our data suggest is highly relevant for young women. We have a unique, efficient, and time-sensitive opportunity to address these gaps by adding gold standard SED assessment via activPAL thigh-mounted accelerometer to the upcoming HHS2 exam. In this contemporary and diverse female cohort (n=4,050, age=36±6 years), we aim to quantify cross-sectional and longitudinal associations of SED with the clinical components of idea cardiovascular health (BP, BMI, total cholesterol, fasting glucose) by using state-of-the-art statistical methods that consider the compositional nature of SED and 24-hr activity and can correct existing, longitudinal self-reported SED via regression calibration. Also, we add HRV and PWV in Pittsburgh and Indiana (n=950) to efficiently study associations between SED and subclinical, mechanistic CVD outcomes in young women. Aim 3 will study interrelationships of SED, APO history, and ideal cardiovascular health to identify novel risk reductions strategies in this high-risk group with limited treatment options. This research will provide critical data to rigorously link SED, CVD risk, and contributing mechanisms and will inform age- and sex-specific SED interventions to test in young women, including those with APO.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY/ABSTRACT The overarching goal of the Visual Sciences CoBRE (VS-CoBRE) is to optimize infrastructure and mentoring to support a sustainable, multidisciplinary, and collaborative research environment at WVU around visual sciences. The establishment of the VS-CoBRE will strengthen the scientific community focused on understanding processes that are essential to visual health with the long-term goal of contributing to novel approaches for prevention and treatment of blinding diseases that disproportionately affect Appalachia's population. West Virginia has the 2nd highest rate of visual disability in the nation, and the incidence is projected to double by 2050. We will achieve our goals by completing the following aims: Aim1, Support Project Leaders (PLs) to achieve independent extramural funding; Aim 2, Promote the fundamental understanding of the visual system through optimizing relevant technical infrastructure; Aim 3, Promote a comprehensive, multidisciplinary approach to impactful visual sciences research; and Aim 4: Implement a rigorous strategy to evaluate the effectiveness of the VS-CoBRE towards promoting a robust center of excellence in visual sciences. With our existing strengths and research infrastructure network, coupled with the initiatives proposed in this VS-CoBRE, including mentoring of our four-promising junior PLs, we are well-positioned to achieve our goal. The creation of a center of excellence provides a venue for integrating efforts across the campus, including activities supported by NIGMS including the WV CTSI and other CoBREs, the clinical enterprise in the Department of Ophthalmology and Visual Sciences, and investigations led by basic biologists in the Departments of Biochemistry, Neuroscience, Biology, and the program in Biomedical Engineering. The VS-CoBRE will coalesce vision scientists around a common theme, enhance the intellectual environment, and cultivate cross collaborations. When the investigators proposed for support by this grant achieve independence by obtaining an R01-equivalent award, we will leverage the VS-CoBRE to recruit additional junior scientists whose work pertains to visual sciences. The core facilities and research infrastructure supported by the CoBRE will also allow our investigators to produce research results that will translate to patients, moving us toward our ultimate goal of reducing the burden of visual impairment and its comorbidities in the Appalachian region, throughout the nation, and worldwide.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT: There are significant differences between men and women in the incidence and severity of late- onset Alzheimer’s Disease (LOAD). After menopause, women are more likely to develop LOAD, and symptoms of the disease including cognitive impairment are more severe. These symptoms are exacerbated by high cholesterol which, at midlife, is a major risk factor for LOAD. There is a substantial gap in our knowledge of how estrogen and cholesterol interact. We propose to examine the role of estrogen and cholesterol in LOAD sex differences by studying male and female cholesterol-fed rabbits – an unconventional but promising model of LOAD. These rabbits show significant sex differences in AD-like pathology, estrogen receptor transcriptional activity and protein expression, and differences in cognition. Cholesterol-fed female rabbits develop beta amyloid (Aβ) deposits more slowly than cholesterol-fed males and eliminating peripheral estrogen by ovariectomy more than doubles Aβ levels, suggesting a protective role for estrogen. We have evidence that a cholesterol diet alters estrogen receptors, significantly increases serum and hippocampal levels of the cholesterol metabolite, 27-hydroxycholesterol (27-OHC), and female cholesterol-fed rabbits remember hippocampally-dependent learning better than cholesterol-fed males. 27-OHC is a well-documented endogenous selective estrogen receptor modulator that may play a role in learning and memory because patients with mild cognitive impairment (MCI) and AD show elevated 27-OHC levels and we have evidence that cholesterol-fed rabbits have elevated 27-OHC and memory deficits. We also have data showing there are sex differences in the transcriptional activity of estrogen receptors and expression of proteins in the presynaptic active zone and postsynaptic density that are higher in female cholesterol-fed rabbits than in males. Our research focus on cholesterol-induced increases in 27-OHC has direct clinical relevance because midlife hypercholesterolemia is a significant risk factor for LOAD and, as noted, 27-OHC is elevated in MCI and LOAD. In three specific aims, we will manipulate estrogen (Aim 1), 27-OHC (Aim 2), and estrogen receptors (Aim 3) in cholesterol-fed rabbits to test the hypothesis that sex differences in AD-like cognitive impairment and pathology are a function of estrogen and can be rescued with estrogen receptor modulation. Using behavioral, electrophysiological, histochemical, and molecular biological techniques, we will determine the mechanisms by which estrogen receptor modulation affects memory, neural function, markers of cholesterol and Aβ processing, and Aβ and tau levels in intact and castrated male and in intact and ovariectomized female cholesterol-fed rabbits. Our expertise in and track record of behavioral, histochemical, electrophysiological, and molecular biological research in cholesterol-fed rabbits makes us a particularly well-suited team to conduct these experiments, further validate this non-transgenic model of LOAD, and positions us to help understand the impact of sex differences on the molecular determinants of LOAD risk and responsiveness to treatment.

NIH Research Projects · FY 2024 · 2021-09

Abstract: Vascular contributions to cognitive impairment and dementia (VCID) is the second leading cause of dementia behind Alzheimer's disease (AD), and is a frequent co-morbidity with AD. Furthermore, the deleterious effect of vascular pathologies combined with AD pathology leads to increased likelihood of dementia. Despite the importance of VCID, little is known about its molecular mechanisms underlying vascular and cognitive dysfunction. This has led the NIH to prioritize studies examining vascular contributions to dementia, and its interplay with AD. Chronic psychosocial stress is a risk factor of VCID. Our preliminary data showing that chronic stress leads to considerable cerebrovascular and neuroinflammatory changes that have similar fundamental changes evident in the progression of AD has led us to focus on this process. Endothelial dysfunction is a critical determinant of vascular disease and predictor of clinical events. Xanthine oxidoreductase (XOR) is a major source of oxidative products (hydrogen peroxide and superoxide) and uric acid. The liver is the site of greatest XOR activity and the main source of circulating XOR activity. As such, XOR can negatively affect the vasculature. Our preliminary data suggest that chronic stress increases XOR activity resulting in cerebrovascular dysfunction and increased inflammation leading to cognitive impairment. Our central hypothesis is that chronic stress elevates hepatic XOR, which is released into the circulation directly causing cerebrovascular dysfunction and the activation of inflammation via a TLR4 pathway resulting in cognitive decline which accelerates dementia/AD pathology. Aim 1 uses a liver (hepatocyte)-and endothelial-specific XOR conditional KO (HXdh-/-) mouse and a liver-specific XOR overexpression tool to manipulate the XOR pathway during 8 weeks of chronic stress and to examine the pathology of VCID. In Aim 2, we will use the global-and- endothelial-specific TLR4-/- mouse, along with physiological approaches to manipulate the TLR4/NF-κB pathway, again in the context of chronic stress and determine VCID pathology. In Aim 3, we will use the increase in XOR activity with chronic stress and switch its bad oxidative products to nitric oxide by supplementing with nitrite and determine its actions on the pathology of VCID. Complementary experiments will also examine the interaction of VCID and AD, by manipulating the XOR pathway (using febuxostat) and determining if we can delay the pathological progression of AD (3xTg-AD mice). Pilot data support this hypothesis. Thus, the overall goal of these studies is to determine the etiology of the stress-related XOR and pro-inflammatory changes in mediating VCID, and its progression to AD pathology. The studies will fill gaps identified by the NIH regarding the need for understanding of vascular contributions to cognitive impairment and dementia.

NIH Research Projects · FY 2024 · 2021-08

Abstract: Despite the significant contributions of transgenic mouse models to our understanding of Alzheimer’s disease, NIA has concluded that these models may be “of poor predictive value in human clinical trials” [RFA- AG-21-003]. As a result, there is a need for new, innovative, non-rodent, mammalian models that better recapitulate the cellular, neuropathological, and cognitive hallmarks of late-onset Alzheimer’s Disease (LOAD). These should include models in which risk factors for LOAD can be systematically induced, and in which cognitive deficits that are the earliest hallmarks of LOAD can be assessed. There is convincing epidemiological evidence that diet and lifestyle are important determinants of cognitive function, but it is unclear how factors such as high cholesterol, obesity, and diabetes increase the likelihood of cognitive deficits. The purpose of the current proposal is to develop, characterize, and validate unconventional, mammalian models that recapitulate the cellular, neuropathological, and cognitive hallmarks of LOAD. The strategy is to feed male and female rabbits a long-term, low-dose cholesterol diet in Aim 1, a high-fat diet in Aim 2, and a diet high in both sugar and fat in Aim 3 to recreate LOAD-like pathology and study the effects of hypercholesterolemia, obesity, and hyperglycemia on learning and memory using a range of increasingly complex tasks – well-understood paradigms that are altered in LOAD patients and we have shown to be affected by dietary manipulations in rabbits. We will also assess the cellular and neuropathological effects of hypercholesterolemia, obesity, and hyperglycemia including their impact on the neurobiology of learning and memory. Compelling preliminary data provide evidence that a short-term, high-dose cholesterol diet, a high-fat diet, and chemically-induced diabetes have deleterious effects on a range of learning and memory tasks. Preliminary electrophysiological evidence shows that feeding a diet high in cholesterol or high in fat can impair a well-known form of synaptic plasticity thought to underlie learning and memory – long-term potentiation. Published and preliminary pathophysiological data show significant diet- induced changes in cellular and neuropathological markers of LOAD. Taken together, these data provide proof of concept, but the levels of hypercholesterolemia and hyperglycemia were high and, although they recapitulated LOAD-like pathologies including beta amyloid accumulation, they also produced off-target pathology. It is therefore important to establish, characterize, and validate the cognitive and pathophysiological effects of milder, more long-term dietary manipulations that induce the types and levels of hypercholesterolemia, obesity, and hyperglycemia that are more clinically relevant. Our expertise in and track record of inducing significant behavioral, electrophysiological, and pathophysiological changes by manipulating diets in adult rabbits makes us well-suited to develop, characterize, and validate these unconventional models of LOAD – models that may represent improved translational potential by better replicating the cellular, neuropathological, and cognitive features of LOAD than current rodent models.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY/ABSTRACT Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly. A common feature in AMD is the early damage of retinal pigment epithelium (RPE), a monolayer of cells between photoreceptors and choroid. RPE controls nutrient transport from choroid blood supply to the outer retina and multiple lines of evidence show that the altered metabolism in RPE is the underlying mechanism for AMD. The long-term goal of this project is to identify key metabolic features of RPE and the roles that they play in AMD. We recently found that besides glucose, RPE preferentially uses proline, an amino acid, to fuel its metabolism. Mutations of enzymes in proline metabolism in humans could cause chorioretinal atrophy or retinal degeneration. The observation that a proline transporter is highly enriched in RPE but not in photoreceptors, further supporting our finding. The proline transporter is a novel risk gene linked to AMD. In our preliminary data, we show that knockout of the proline transporter blocks proine utilization and impairs visual function. The objective of this proposal is to study proline metabolism in RPE and its role in retinal degeneration. We plan to rigorously investigate the role of proline in healthy and diseased RPE using advanced tracer methodology, mass spectrometry, in vivo infusion, live imaging, and proline transporter knockout mouse models. The outcome of this research will be to establish a conceptual framework for RPE metabolism that describes how RPE uses nutrients to maintain metabolic homeostasis with the outer retina. This new knowledge will provide the basis for understanding the mechanisms of AMD and lay the foundation for developing new treatments.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY Microbial infections are a major cause of infant mortality worldwide. For particularly vulnerable populations such as pre-term and low birthweight babies, the risk of invasive infections further escalates. The neonatal period is defined by a distinct, often described as immature, immune system. Many features of a protective host response to infection are deficient as compared with older children and adults. Our laboratory has identified that expression of the immune suppressive cytokine interleukin (IL)-27 is elevated in human and murine neonates. Other recent studies have shown IL-27 to be a biomarker for early onset neonatal sepsis. This suggests that elevated IL-27 may represent a risk factor and when further increased during bacterial challenge, compromise the host immune response. The overall premise of the current proposal is that IL-27 is a host molecule that represents a target for immune intervention to improve the host response and reduce susceptibility to bacterial infection early in life. We present strong evidence in a mouse model that the absence of IL-27 signaling translates to increased survival, improved weight gain, and enhanced clearance of bacteria during neonatal sepsis. To advance our knowledge of how IL-27 regulates the immune response during neonatal sepsis, we need to identify the complete repertoire of cell types responsible for IL-27 production, understand how these population may change over the course of infection, and further define their functionality. We will address this gap in understanding using an IL-27 reporter mouse that expresses a fluorescent protein under control of the IL-27p28 promoter. Using whole-animal imaging of the reporter mouse coupled with luminescent bacteria, this will allow us to identify IL-27 producers, sort them for further functional analysis, and correlate their presence in infected tissues with the bacterial burden. We also seek to understand cellular signaling pathways required for IL-27-mediated suppressive activity and compromised control of the bacterial burden. We hypothesize that signal transducer and activator of transcription (Stat)-3 signals downstream of IL-27 receptor binding to interfere with lysosomal trafficking and acidification. The net result is compromised bacterial clearance. Lastly, a primary objective is to investigate the outcomes of antagonizing IL- 27 during neonatal sepsis with the aim of establishing an immunotherapeutic approach for an infectious disease for which we can currently only offer antibiotics and supportive care. Antibiotic resistance confounds our reliance on this approach. Administration of a neutralizing antibody conjugated to a fluorescent tag will allow for visualization of tissue penetration in real time and directly correlate the presence of the antagonist with control of bacterial growth. At the completion of this project, we expect to have performed preclinical validation of a promising immunotherapeutic approach to improve immunological responses and susceptibility to infection disease in newborns, as well as provided an enhanced understanding of how IL-27 regulates host immunity and interactions with bacterial pathogens during neonatal sepsis.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY The objective of this application is to determine how tissue nonspecific alkaline phosphatase (TNAP) enzymatic activity maintains cerebrovascular function within the neurovascular unit (NVU). Brain microvascular endothelial cells (BMECs) comprise the cerebral microvasculature and serve as the structural foundation of the blood-brain barrier (BBB) and. Increased permeability and diminished integrity of BMECs are two common mechanisms through which cerebrovascular function is compromised in human disease. TNAP is a highly enriched enzyme in cerebral microvessels whose function in brain BMECs is poorly understood. Our preliminary data demonstrate that TNAP activity stimulates a novel signaling mechanism which protects against the loss of cerebral microvascular integrity and permeability. These intriguing findings led us to propose the central hypothesis that TNAP maintains NVU homeostasis during cerebral ischemia by preserving BBB integrity. We will utilize mice with a VE-cadherin-Cre driven conditional deletion of TNAP in endothelial cells (VEcKO) and its wild type littermates to interrogate the role of TNAP in BMECs. We will compare responses in young (4-5 months) and aged (18-20 months) mice to assess the age-dependent effects of brain endothelial TNAP on indices of brain endothelial barrier permeability combined with vascular network analysis and functional behavioral outcomes. Aim 1 will elucidate the contribution of brain endothelial cell TNAP to NVU dysfunction in ischemic stroke in young mice. We will employ the transient middle artery occlusion model to assess quantitative differences in cerebrovascular outcomes and behavioral indices. Aim 2 will determine the impact of brain endothelial cell TNAP on age-dependent impairment of the NVU. This aim will assess the impact TNAP on BMEC function in aging and the putative age-dependent interactions in ischemic stroke. Aim 3 will determine how the TNAP-Rho associated kinase (ROCK) pathway regulates the barrier function and whether pharmacological inhibition of the ROCK pathway protects against the loss of barrier integrity and functional deficits associated ischemic stroke in both young and aged mice. Taken together, the studies in this proposal will delineate a novel mechanism through which brain endothelial cell TNAP enzyme activity preserves NVU function in ischemic stroke and improves functional recovery post-stroke. The overall results will contribute to our limited understanding of the basic biology of TNAP’s role at the BBB and its contribution to NVU homeostasis in human health and disease.

NIH Research Projects · FY 2025 · 2021-01

PROJECT SUMMARY Pertussis is a respiratory disease caused by the obligate human pathogen Bordetella pertussis. Two generations of pertussis vaccines have been developed and licensed: whole cell pertussis (DTP) and acellular pertussis (DTaP/Tdap). Pertussis was thought to be a disease of the past but has recently re-emerged. The number of cases of pertussis in 2012 was 48-fold over the lowest year on record (1976), which was also a 50-year high. While the increase of pertussis has multiple potential reasons, epidemiological studies clearly suggest that the duration of immunity of both DTaP and Tdap wanes quickly each year after a booster, and regresses to non- protective levels in humans. Each dose of whole cell vaccine contains hundreds of antigens, of which numerous are immunodominant. Whole cell vaccines also induce T helper 1 and 17 (Th1/Th17) cellular immune responses. On the other hand, acellular vaccines focus Th2-mediated humoral responses exclusively to pertussis toxin, filamentous hemagglutinin, pertactin, and fimbriae. We aim to develop a vaccine that would induce Th1 responses and include a greater number of antigens than acellular vaccines. mRNA vaccines provide a platform that can be easily modified for the targeted antigen/pathogen and they induce Th1 responses. mRNA vaccines encode the antigen, which once expressed, results in immunity mediated by Tfh responses. We used an mRNA platform to screen antigens of B. pertussis and identified a protective multivalent formulation (mRNA-DTP10; 8 pertussis antigens with diphtheria and tetanus antigens) in a murine challenge model. In this project, we will further extend our studies and investigate the correlates of protection of the mRNA-pertussis vaccine in the murine model with longevity studies and examine DTaP prime / mRNA-boost effects (SA1). Next, we will utilize the coughing rat model of pertussis with whole body plethysmography to compare mRNA, whole cell, and acellular immunity for protection against cough (SA2). We will study the mRNA pertussis vaccine in immunogenicity and challenge experiments using the baboon model of pertussis (SA3). Lastly, we will aim to develop a suite of assays to phenotype antibodies produced by mRNA pertussis vaccines and bridge each of the pertussis models, which will facilitate clinical development. Through these studies, we aim to characterize the mRNA-pertussis immunity and develop a clearer understanding of how this vaccine platform can be used to overcome “complex or difficult” pathogens that employ numerous virulence factors. We expect the data acquired in each aim will result in a deeper understanding of pertussis immunity as well as illuminate the mRNA platform for bacterial vaccines.

NIH Research Projects · FY 2024 · 2020-09

Open-angle glaucoma (OAG) is managed primarily with topical medical therapy, with laser and surgical interventions reserved for those in whom medical therapy is ineffective, intolerable, or inappropriate. The effectiveness of medical therapy is limited by patient adherence with therapy; inadequate adherence has been extensively documented. Selective laser trabeculoplasty (SLT) has comparable efficacy to preferred first-line prostaglandin analogues, is very safe, and obviates the need for daily medical therapy in most patients when applied as primary therapy. A recent randomized trial demonstrated superior glaucoma outcomes (less progression, fewer surgeries required) in newly diagnosed OAG patients receiving primary SLT vs. medical therapy, providing the evidentiary basis for a paradigm shift that is already underway in which SLT supplants medical therapy as the preferred first-line treatment for OAG. SLT is largely performed as first described by its inventor; little exploration of dose-response has been undertaken. An intriguing data set from Italy (with significant weaknesses and limitations) suggests that low energy SLT repeated annually is far more effective than standard SLT repeated as needed when its effect wanes, in delaying or preventing the need for topical medical therapy. This finding is consistent with the limited data exploring the dose-response relationship between SLT and intraocular pressure (IOP) reduction. Further, it is biologically plausible that ongoing health maintenance of trabecular meshwork (TM) function with proactive annual low energy SLT would better preserve the TM's long-term health and function than a repeated cycle of SLT, progressive TM re-impairment by the glaucoma process, loss of IOP control, and repeat SLT. Our proposal describes a multi-center clinical trial to answer two key questions: 1) Is primary low energy SLT as effective as primary standard energy SLT in newly diagnosed and treatment-naïve patients with mild-moderate OAH or high-risk ocular hypertension?; and 2) Does annual low energy repeat SLT more effectively delay or prevent the need for medical therapy compared to standard SLT repeated as needed when its effect wanes and IOP rises? Participants will be randomized to initial standard SLT or initial low energy SLT with the possibility of a single repeat SLT as needed in the first year of the study. The first primary outcome will be 12-month survival where failure represents the need for repeat SLT to achieve/maintain protocol-specified target IOP. At Month 12, all participants who remain medication-free will be re-randomized to undergo repeat SLT either as needed when IOP exceeds target IOP (at initially randomized energy) or to annual low energy SLT irrespective of IOP. The second primary outcome will be 42-month medication-free survival in subjects who were medication-free at Month 12. Our study seeks to clarify the optimal way to utilize SLT with the ultimate goal of maximizing long- term SLT responsiveness and medication-free survival. If successful, our results will shape the course of treatment for most newly diagnosed and treatment-naïve patients with mild-moderate OAG or high-risk OHT.

NIH Research Projects · FY 2024 · 2020-09

SUMMARY: Over 60% of children experience severe stress and are exposed to traumatic events including interpersonal violence, sexual abuse, accidents and injuries – adverse childhood experiences – but there is a mismatch between their exposure to these experiences and the prevalence of subsequent psychopathology. This mismatch, in which most children who experience traumatic events do not show psychopathology, may result from resilience to the events, a lack of diagnosis, or forgetting about the experiences. Resilience to adverse events involves responding with minimal distress or an early and effective return to normal levels of function. Forgetting about traumatic events in the very young – referred to as infantile amnesia – has been associated with critical periods in development involving the formation and strengthening of perineuronal nets surrounding neurons in specific areas of the brain related to memory formation for highly stressful events. Disrupting these perineuronal nets may extend or renew critical periods and help allow the memory of adverse experiences to be erased. Using a new model of hyperarousal in young rats to model adverse childhood experiences, we will determine the ontogeny, mechanisms, and treatment of hyperarousal. Our overarching goal is to understand the hyperarousal that results from stressful events. We will test the hypothesis that resilience to and forgetting about learning-induced hyperarousal is a function of perineuronal nets that form and strengthen during development around neurons in the circuits underlying associative learning. To test this hypothesis, we focus on three specific aims: (1) Characterize the ontogeny of hyperarousal and determine the underlying neural mechanisms, (2) Determine behavioral strategies to “treat” or mitigate hyperarousal in young rats and delineate the neural mechanisms involved, and (3) Determine the role of perineuronal nets in hyperarousal and its treatment. We will conduct a series of experiments in which we characterize hyperarousal in young rats, determine treatments, and then manipulate perineuronal nets before acquisition or extinction of aversive associative learning to determine whether we can manipulate critical periods to impair the development or facilitate the forgetting of hyperarousal as well as the conditioned emotional responding to cues associated with adverse events. The proposed experiments constitute a concerted effort to fill an important gap in our understanding of the developmental trajectory of hyperarousal that occurs in children following adverse events – an area of growing concern as the incidence of interpersonal violence, accidents and injuries to children continues to escalate both in the United States and abroad. We will focus on mechanistic studies that reveal the underlying neural processes, the role of perineuronal nets, and elucidate age-specific behavioral and pharmacological treatment strategies.

NIH Research Projects · FY 2025 · 2020-09

West Virginia (WV) bears long-standing physical, psychosocial and fiscal characteristics, limited resources and a restricted economy, that have significantly impacted the health of its residents. The West Virginia Pediatric Clinical Trials Network (WVPCTN), as a Clinical Site of the ECHO IDeA States PCTN (ISPCTN), has catalyzed emerging solutions to address some of these burdens by fulfilling its mission. Specifically, the mission of the WVPCTN has been two-fold - bringing clinical trials to our at risk population, and building professional capacity for clinical research. Our WVU site has been and continues to be well positioned to be a successful Clinical Site due to a combination of structural and human resources that were already in place when the ISPCTN was initiated in 2016, and the background and expertise of the then three PIs – Drs. Saul, Pyles and Cottrell. The addition of the funds and resources provided by ISPCTN led to an acceleration of WVU’s capacity to actively participate in clinical trials by allowing us to create a dedicated Pediatric Research Unit (PRU)/Children’s Hospital Research Consortium (CHRC) and introduce a group of junior, less experienced clinical trial investigators, into both ISPCTN and non-ISPCTN trials. The effectiveness of this approach has been demonstrated through an increase in faculty participation in ISPCTN activities, a strong recruitment record, and an increase in the number of non-ISPCTN Pediatric trials at WVU. The WVPCTN is now actively growing an experienced investigator pool and a group of mentee investigators, all enhanced by the resources and expertise of the West Virginia Clinical Translational Science Institute (WVCTSI). These features and resources, together with strong and sustained partnerships throughout the state, and the multiple underlying conditions of our citizens, continue to uniquely position us to be successful as an ISPCTN Clinical Site and work to propose new trials for the network. This proposal for WVPCTN3 will further add to a long-standing commitment to Pediatric-focused initiatives across the state. When funded, it will continue to position us to fully leverage additional funds that support Pediatric research infrastructure, investigator development, and community input to provide access to cutting edge trials which will positively impact the health of the children of WV.

NIH Research Projects · FY 2025 · 2020-08

Project Abstract Many regions of the brain including the cortex, hippocampus, basal ganglia, and limbic structures are highly enriched with synaptic zinc. Synaptic zinc (as Zn2+) is loaded into presynaptic vesicles by zinc transporter 3 (ZnT3), where it is coreleased with glutamate during synaptic transmission. Synaptic zinc can modulate many fundamental aspects of synaptic signaling. ZnT3 KO mice (which lack synaptic zinc) display a range of cognitive and sensory impairments and demonstrate behavioral deficits associated with autism and schizophrenia. Mounting evidence from human populations shows that mutations in certain zinc transporters are linked with major neurological disorders such as schizophrenia. Together, these findings strongly suggest that synaptic zinc signaling is important for neuronal processing. Our laboratory is focused on understanding how brain-specific zinc transporter proteins control zinc release and clearance at synaptic terminals, how different types of brain cells utilize zinc during synaptic transmission, and how the effects of zinc regulate the activity and structure of individual synaptic connections in the brain. Together these approaches will allow us to answer fundamental questions concerning the role of synaptic zinc in brain function and provide new mechanistic insights into endogenous mechanisms that shape synaptic and neural processing. The overall vision for project over the next five years is to understand the dynamics of endogenous synaptic zinc in the brain: how it is released, how it is cleared, how it interacts with pre- and postsynaptic receptors, how it supports synaptic structure, and how it is regulated by zinc transporter proteins. Because zinc is a fundamental aspect of cellular function, understanding the mechanisms underlying its actions at synapses will not only improve our understanding of cellular processes important in the brain, but will also pave the way toward new targets in the treatment of pathological conditions.

NIH Research Projects · FY 2024 · 2020-07

PROJECT SUMMARY/ABSTRACT The overall goal of this project is to elucidate the role neutrophils, both peripheral and meningeal, play in CNS homeostasis. Specifically, how neutrophil activity and location may influence the molecular landscape of neurons in learning and memory. During the mentored phase of this project, I will focus on characterizing the neutrophil population within the meninges, the location of these cells in the meninges, and determine whether the meninges act as a non-medullary source of neutrophils (Aim 1). During this period, I will also determine the specific role these cells play in modulating learning and memory in mice (Aim 2). The completion of these aims will help me develop skills in bioinformatics, establishing slice cultures and gain a rudimentary working knowledge of electrophysiological methods. These techniques will be of further use to me in the independent phase of this grant. During the independence phase, I will investigate elucidate the role of peripheral and meningeal neutrophils in different learning paradigms (Aim 2). Upon completion, this project will elucidate new neuromodulatory function of neutrophils and identify the meninges as a source of these cells within the CNS. These results will greatly inform our understanding of the role these cells play in injury and neurodegeneration.

NIH Research Projects · FY 2025 · 2020-05

ABSTRACT/SUMMARY Head and neck carcinomas (HNC) are some of the most challenging cancers to effectively treat (five-year survival rates for some HNCs are as low as 25%); recurrence rates range from 8% to 43%. The number of new cases is rising due to the current, near epidemic of HPV-associated HNCs. Unfortunately, most treatments are associated with significant morbidity due to damage of sensitive structures in the region (spinal cord, salivary glands, parotid gland, esophagus, carotid arteries and thyroid gland). HNCs are treated with surgical excision, radiation therapy, or a combination of methods. Selection of therapy is based upon assessment of tumor location, size, proximity to bone, amount of infiltration into surrounding tissues and spread to regional lymph nodes performed with advanced imaging methods. Perhaps the most promising of these methods is metabolic-based imaging, specifically PET, often utilizing FDG. The non-optimal spatial resolution and fixed geometry of whole body scanners, however, limits PET in fulfilling its promise in this role by inhibiting the ability to accurately detect small tumor masses in lymph nodes, and in quantifying the size and nature of the primary tumor. Thus, there is an unmet need for improved PET/CT scanner technology to enhance treatment planning of HNCs. To address this opportunity, we propose the creation and testing of a lower-cost, flexible geometry, high-resolution PET/CT system (approaching the spatial resolution of pre-clinical PET scanners), called HNPET/CT, designed specifically for the imaging of the head and neck region. It will consist of a novel pair of large area, immersion-cooled, PET detectors and a cone beam CT (CBCT) scanner mounted on a rotating gantry whose geometry can be tailored to patient size and anatomy to be scanned. To capitalize on HNPET/CT’s high-resolution images, an image segmentation method that utilizes both CBCT and PET images will be developed and tested. We plan to explore the potential utility of HNPET/CT for enhancing therapy planning (surgical and radiation). This assessment will first be performed with anthropomorphic phantoms and then in a limited human trial. In addition to the novel detector design and adjustable geometry, HNPET/CT will introduce a new capability to the treatment planning of HNCs not yet broadly employed by end users in the clinical. It could enhance current planning techniques (reduce treatment margin size, for example), enable the effective application of advanced methods (dose painting, for example), and perhaps inspire development of new, more effective personalized treatment strategies that require high resolution, multi-modality imaging. The lower cost of the system promises the introduction of cutting-edge, image-guided treatments to under-served populations in areas treated by small or private clinics that often cannot afford such technology. This project will be performed by a multi-disciplinary academic industrial partnership joining the Departments of Radiology, Head and Neck Surgery and Radiation Oncology at West Virginia University, and Xoran Technologies, LLC.

NIH Research Projects · FY 2024 · 2020-01

Abstract Congenital heart disease (CHD) affects approximately 1.2% of children and is the leading cause of birth defect- related deaths. Single ventricle heart disease (SVHD) is a severe form of CHD, with high morbidity and mortality. These patients require multiple palliative surgeries, culminating with a total cavopulmonary anastomosis. Despite considerable improvement in the survival of patients with SVHD, there is increasing morbidity and mortality over time. It remains unclear why some SVHD patients fail their surgical repairs while others remain relatively well. Clinicians often rely on 2-dimensional (2D) images acquired from echocardiograms, catheterizations, or cardiovascular magnetic resonance (CMR) exams to assess SVHD patients and qualitatively choose the optimal surgical repair. The 2D images, however, lead to a suboptimal understanding of the complex 3D spatial relationships and hemodynamics, and limit efficient decision making. To address this deficiency, a free-breathing sequence is developed to acquire 3D cine CMR images of the heart and great vessels in 10 minutes. The 3D block of data will be used to generate a patient-specific pulsatile heart model. This heart model will be used to simulate the patient cardiovascular system with a lumped- parameter model. The lumped-parameter model will be used to simulate different surgical repairs and quantitatively choose the most optimal repair for each patient. We expect that this platform rationalizes surgeons' decisions for the best surgical approach and improves the survival rate of patients with SVHD.

NIH Research Projects · FY 2025 · 2019-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The Cell & Molecular Biology and Biomedical Engineering Training Program (CBTP) at West Virginia University is designed to provide advanced research training in cell biology, molecular biology and biomedical engineering to PhD students interested in pursuing careers as independent scientists. CBTP draws upon the existing high level training environments in the biomedical sciences PhD programs in the WVU Health Sciences Center and the PhD program in the Department of Chemical and Biomedical Engineering in the Statler College of Engineering and Mineral Resources, which recently developed a PhD program in Biomedical Engineering. The CBTP program aims to develop and sustain interdisciplinary graduate training in the biomedical sciences and biomedical engineering to prepare a biomedical workforce to address complex problems associated with human health and disease. The goals are to enhance scientific training, prepare trainees to tackle problems using multidisciplinary approaches and work in teams. The program plan has science-related objectives to enhance breadth and depth of scientific knowledge and increase the exchange of ideas between disciplines. The program plan aims to enhance professional and career development by expanding career awareness, providing training in professional skills and promoting experiential learning. The program has additional objectives to promote the ethical conduct of research, and rigor and reproducibility in science. Key activities to promote career awareness include Exploring Careers Symposia, a Non-academic Careers seminar series, and discussions between preceptor panels and trainees to provide transparency about the current biomedical workforce and training outcomes. Professional skills training will occur in stand-alone workshops and in workshops incorporated into other activities, e.g., program retreats or symposia. The program also devotes time to responsible conduct of research discussions between trainees and preceptor panels. The program plans to support six trainees per year and to make appointments for two years, with review at the end of the first year, prior to reappointment for a second year. The intended outcome of training is the development of students with an appreciation for multidisciplinary science, broader approaches to problems, and the ability to work collaboratively. Training will enhance professional skills and prepare students to promote the ethical pursuit of science.

NIH Research Projects · FY 2025 · 2019-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The Research Training Program in Behavioral and Biomedical Sciences (BBS) at West Virginia University is designed to provide advanced research training at the interface of behavioral and biomedical sciences to PhD students planning careers as independent investigators. The BBS Program draws on existing, high-level training in the Biomedical, Psychology, Biology, and Public Health PhD Programs and provides considerable added value through curricular offerings (including behavioral neuroscience methods, biostatistics, bioinformatics, and epidemiology), a highly interactive journal club and an extensive seminar series. Specific interdisciplinary training and experiential activities are provided to foster scientific leadership, communication skills, responsible conduct of research, professional development, advanced training in methodological rigor with attention to reproducible research, and cognizance of issues related to human health, physiology, and disease. Structuring the BBS Program to leverage the strengths of the Biomedical, Psychology, Biology, and Public Health PhD Programs ensures an experienced and productive preceptor group, a wide range of research training opportunities, and a highly qualified applicant pool. The BBS Program provides a proven vehicle for training the next generation of scientists with the intellect, knowledge, experience, resilience, and courage to address complex research problems with significant

NIH Research Projects · FY 2024 · 2018-09

Abstract: As the national toll from the opioid epidemic continues to worsen, West Virginia leads the nation in overdose deaths, with 52 deaths per 100,000 population, and the economic impact of the opioid epidemic on West Virginia is estimated at around $1 billion. The impact of the opioid epidemic is not limited to West Virginia; nationally, more than 63,000 overdose deaths occurred in 2016. Unfortunately, despite the growing necessity of treatment for Opioid Use Disorder (OUD), most patients do not respond to the current standard of care. The success rate of patients initially stabilized on the gold standard of treatment, Medication Assisted Therapy (MAT) in conjunction with psychosocial interventions, is currently less than 50%. Those who fail the gold standard have a high risk of overdose or other complications. Due to the growing number of OUD patients and the limitations of current treatment, it is clear that new treatment options are needed to confront this problem. Deep Brain Stimulation (DBS) has been used for many years to successfully treat movement disorders such as Parkinsons, and has shown promise in investigations for other mental disorders. We hypothesize that implanting the DBS device in the nucleus accumbens region of the brain will modulate the reward and behavior self- regulation networks, which will decrease opioid cravings. This proposal aims to test safety, feasibility, and tolerability in an initial cohort of 4 participants during the UG3 phase. This will allow for an evaluation of safety and tolerability of DBS in participants with treatment-refractory OUD and a history of life-threatening complications secondary to opioid use. If the UG3 phase is successful following intensive monitoring of the 4 participants, the UH3 phase will consist of a randomized, controlled proof-of-concept study with 16 participants. These participants again will have treatment-refractory OUD and a history of life-threatening complications secondary to opioid use. During the UH3 phase, the mechanism of DBS impact on OUD will be investigated through both neuroimaging and measurement of executive function.

NIH Research Projects · FY 2025 · 2018-08

PROJECT SUMMARY/ABSTRACT The efforts described in this second phase CoBRE application will support the Center of Excellence within the West Virginia University Health Sciences Center (WVU HSC) that focuses on studies of the tumor microenvironment (TME), designated the TME CoBRE. The overarching critical need for continued emphasis in this area is driven, in part, by that fact that cancer mortality is a significant health disparity in the Appalachian region, specifically in West Virginia. We are building on a successful Phase I, where we had two graduates with R01s, a major national foundation grant, a NSF grant, an R21 and our cores were awarded two S10 awards. In total we had 11 Project Leader (PL) awards as PI, and 8 as a Co-I. The Center has grown to over 20 faculty, including a number of faculty who are potential PLs, with their teams publishing over 140 papers since initiation of Phase I. Laboratories supported, in part, by the Phase I TME CoBRE, have engaged in the training of ~60 students further amplifying the impact of the NIGMS investment on workforce development in parallel to CoBRE PL success. Herein, we provide details for the strategy to continue to develop careers of promising junior scientists and recruit additional investigators to study the biology of, and novel therapeutic approaches that will benefit from, a mechanistic understanding of the diverse TME. The five highly translational projects in Phase II focus on the microenvironment of different tumor types, including cancers initiating in the bone marrow, gastric system, breast, and brain. The administrative core will manage the overall budget and provide assistance in annual reporting and submission of extramural applications of all CoBRE Project Leaders. In addition, this core will provide oversight of mentoring, which includes two primary advisors for each investigator as well as an external advisor, and a network of previous CoBRE graduates and the Director of Core Resources. The investigators will be supported by two research cores that leverage past and current CoBRE and IDeA support. Single cell analysis capability in the existing flow cytometry core will aid to it investigations into genomics of single cells. The Imaging Core will support PLs with cutting edge technologies including microbeam irradiation, microCT, pre-clinical MRI, and real time microenvironment imaging of pO2 and pH. Lastly, we will continue to administer a pilot project program to recruit new junior faculty to the TME CoBRE, as we saw five of six pilot grant awardees eventually become PLs, and we currently have a robust number of junior investigators (#5) may become PLs as well. The investigators are well integrated into established programmatic areas in the West Virginia University Cancer Institute that meet every other month for focused discussion on therapies that leverage, and the biology of, the TME. The mentoring atmosphere, core facilities, and significant institutional support that are central to this effort will continue to provide a rich environment to nurture investigator independence and success around the critical scientific area of TME.

NIH Research Projects · FY 2025 · 2017-05

Project Summary Stroke - like many brain diseases - is clearly associated with aging and a plethora of age-related co-morbid conditions, including cognitive decline, Alzheimer’s disease (AD), AD related dementias, multi-infarct dementia, cardiovascular disease, cerebrovascular disease, hypertension, hypercholesterolemia, immune suppression, metabolic syndrome and obesity, sleep deprivation, and depression. However, age-related co-morbid conditions, as a variable in stroke occurrence, severity and long-term recovery is seldom studied experimentally, but is one of the identified factors in the lack of progress in the discovery of new therapies for acute stroke brain damage. To optimize the impact of our research on the discovery of new preventatives, acute treatments and rehabilitation methods for stroke, we need to train the next generation of stroke researchers to assess stroke in the context of the affected patent population: those who are elderly and have multiple co-morbid conditions. The Stroke and Alzheimer’s Disease Related Dementias Predoctoral Training Program will formalize and standardize our already strong training programs in the biomedical sciences and focus on stroke research. A number of innovative aspects of this training program are semester-long didactic courses in “Stroke” and the “Neurobiology of Aging”, a clinical immersion in our WVU Stroke Center, a in Stroke Journal Club, an Associate Scholars Program, and experience in community engagement. The proposed training program will select the best PhD students from the participating Biomedical Sciences Training programs at the West Virginia University (WVU) Health Science Center (WVU HSC) and will prepare them with the skills, knowledge and acumen needed for a successful career in stroke research. The specific training for each of 8 mentee will be tailored based on their annually updated Individualized Development Plan (IDP), and an “Exploring Career Paths” Program will help prepare them for their chosen career. Program training is expected to last 2-to-3 years. This pre-doctoral training program will create a new generation of young scholars who can address the need for innovative stroke research for the citizens of West Virginia and the nation.

NIH Research Projects · FY 2025 · 2017-04

PROJECT SUMMARY/ABSTRACT The long-term goal of our research is to identify mechanisms that contribute to the maintenance of photoreceptor outer segments and vision. In retinal degenerative blinding diseases, loss of ciliated photoreceptor outer segments precedes the death of photoreceptor neurons. Therefore, a thorough understanding of the mechanisms behind the stability of the cilia is needed to uncover molecules required for photoreceptor survival. The specific goal of this proposal is to identify the importance of post-translational modifications of tubulins in the maintenance of cilia and photoreceptor function. Altered tubulin modifications are a known cause of human blindness. We will investigate the need for a small GTPase that belongs to the ARF-like family of proteins, ARL13B, and its role in photoreceptor ciliary maintenance, tubulin modifications, and function. Our studies will investigate whether altering tubulin modifications will protect photoreceptors and rescue visual response in various genetic models for retinal degenerative diseases. We will use a combination of unique animal models, cell culture models, and in vitro biochemical analyses to comprehensively address the requirement for tubulin modification and their regulation in ciliated photoreceptors. Our proposed studies are aligned with the Retinal Diseases Program of the NEI to “Elucidate the molecular mechanisms that lead to photoreceptor degeneration, including signal transduction pathways, defects in protein folding, ciliogenesis, functional compartmentalization, or trafficking, and translate these molecular footholds into therapies for Mendelian and complex diseases.” The findings from the proposed studies have clinical implications, such as therapy for inherited retinal diseases that lead to blindness.