Medical University Of South Carolina

NIH Research Projects · FY 2026 · 2024-02

Abstract The Ehlers Danlos Syndromes encompass a group of heritable connective tissue disorders with a complex phenotype affecting multiple body systems. Among these, the hypermobile subtype of EDS (hEDS), is the most common type of EDS affecting 1 in 500 individuals. The disease phenotype is characterized by hypermobile joints that lead to musculoskeletal defects, skin manifestations, tissue fragility, and multiple comorbidities. The healthcare burden associated with hEDS is substantial, with patients often requiring care from a variety of medical specialists, frequent surgeries and hospitalizations, management of debilitating chronic pain, and consistent physical and occupational therapy. Despite well-documented inheritance, the genetic causes of hEDS remain unclear, and diagnosis relies on clinical criteria, resulting in an average 14-year delay in diagnosis. There are currently no direct treatment options for hEDS other than symptom management. Recent findings from our group have highlighted the crucial role of a gene family of trypsin-like serine proteases in hEDS. This proposal will build on these discoveries by exploring the molecular, cellular, and additional genetic mechanisms that contribute to hEDS pathogenesis and connective tissue homeostasis. We will explore how proteins encoded by hEDS genes regulate collagen fibrillogenesis (Aim 1), investigate their contribution of mast cells to altered extracellular matrix (Aim 2), and perform genome wide association studies (GWAS) to reveal common variants associated with hEDS and connective tissue biology (Aim 3). By employing our unique and novel mouse models and leveraging our extensive clinical genetic registry, we aim to gain novel insights into the pathogenesis of hEDS and its related comorbidities. From these discoveries, we aim to accelerate the development of targeted therapies for hEDS, create more accurate molecular diagnostic tools and ultimately alleviate the burden on patients and the healthcare system.

NIH Research Projects · FY 2026 · 2024-02

The primary objective of the Raise Awareness In Students Early (RAISE) in Oral Biology Program is to provide short-term research experiences and training for talented undergraduate students in the area of oral biology. The Medical University of South Carolina (MUSC) RAISE Program will 1) recruit and engage undergraduate students who demonstrate ambition and determination to pursue an oral biology-related career; 2)provide the knowledge and skills that will prepare students for advanced degree admission and a biomedical career in oral biology, and 3) enhance the research workforce by nurturing the academic and professional growth of students in the field of oral and systemic health. The 10-week program will recruit nine students per year. Special consideration will be given to students from non-research-intensive South Carolina higher education institutions. Students will conduct a biomedical hypothesis-driven project under the guidance of a faculty mentor whose research program is aligned with the National Institute of Dental and Craniofacial Research (NIDCR) scientific priorities. The program will leverage the expertise of faculty, many of whom are established mentors and investigators on NIDCR funded fellowship/training and research grants. Dr. Yilmaz, DDS, PhD and Dr. Westwater, PhD with their complementary backgrounds in academic mentoring bring forth a successfully integrated undergraduate oral biology program encompassing basic science, clinical knowledge, and team-based leadership. In addition to hands-on research training in oral biology, students will receive responsible conduct of research training, professional development, and networking opportunities. Activities will include an authentic research experience; a specialized lecture series; formal and informal discussions on the core principles of rigor and reproducibility; and a journal club focused on the connection between oral health and systemic conditions. Trainees will gain an appreciation for the translational relevance of their studies through inter professional experiences in a clinical setting. The program will culminate with formal presentation at the MUSC Summer Undergraduate Research Symposium, a one-day conference in which students present their research to their peers, primary mentors, and research team advisors. Students will be encouraged to build both social and academic networks. Each student will be paired via the RAISE Alumni Network with a student, who will provide professional and social support while on the MUSC campus. After graduating from the RAISE Program, students will be enrolled in the RAISE Alumni Network. Inclusion in this network will facilitate post-program career planning and will increase the likelihood that students will select long-term careers in oral biology-related fields. In turn, we expect our alumni to become part of the research workforce who will focus on improving oral health and well-being for all individuals across the lifespan.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY Transposons, also known as transposable elements (TEs), are parasitic mobile DNA elements that, when re- activated, can introduce regulatory or protein coding changes at a new site in the genome. Previous studies showed TE activation was primarily in the germline and was thought to be overall inactive in differentiated somatic cells and tissues. However, transposon re-activation has been observed to occur frequently in the gastrointestinal tract, where it has been associated with diseases such as Crohn’s, colitis, and cancer. The reasons for increased TE activity in the gastrointestinal epithelium are still poorly understood, however. Interestingly, we have evidence of a novel interaction linking epithelial adherens junctions with transposon regulation. The adherens junctions, comprised of E-cadherin and p120 catenin as the core adhesion complex, act to stabilize cell-cell contacts and are frequently compromised in diseases of the colon epithelium. Preliminary studies in the lab have uncovered an interaction of the adherens junctions with PIWIL2 (PIWI-like mediated gene silencing 2). PIWIL2 is an endoribonuclease that is responsible for transposon silencing and germline maintenance by interacting with piRNAs (PIWI-interacting RNAs) to target TE RNA transcripts for degradation. We found co-localization of PIWIL2 with E-cadherin, p120 catenin, and PLEKHA7 which is an adherens junction specific protein marker in well-differentiated colon epithelial cells and tissues. Furthermore, our data show that disruption of the adherens junctions through depletion of E-cadherin and PLEKHA7 results in mis-localization of PIWIL2 to the cytoplasm or nucleus. E-cadherin depletion also increases the levels of the LINE1 transposon, which is the only autonomously active transposon in humans. Additionally, through RNA-sequencing, we have shown a specific subset of piRNAs are depleted when PLEKHA7 is depleted. From these data, we hypothesize that PLEKHA7 recruits PIWIL2 to the adherens junctions to suppress transposon activity in differentiated cells to maintain the integrity of the epithelium by preventing DNA damage caused by transposons. In Aim 1 we will examine how PLEKHA7 regulates the formation of a functional PIWIL2-piRNA complex, and use shRNA knockdown and CRISPR/Cas9 to deplete components of the complex and examine effects on its functionality by using retrotransposition and genomic integrity assays. The experiments proposed in Aim 2 will dissect the mechanism of PIWIL2 recruitment specifically to the mature apical adherens junctions in well-differentiated colon epithelial cells and examine whether disruption of this mechanism is critical in diseases such as inflammatory bowel disease (IBD), including Crohn’s and colitis, as well as colon cancer using patient tissues and an organoid system. Completion of the proposed study and training plan will further develop my technical skills, research independence, and clinical knowledge, and will position me to become a successful independent investigator.

NIH Research Projects · FY 2026 · 2024-01

Abstract: Approximately 53,000 people in the United States are diagnosed with oral cancer yearly, and the 5- year survival rate remains poor due to late diagnosis and intervention. Recent evidence has suggested periodontal disease as a potential risk factor, and it is also associated with poor outcomes. Interestingly, periodontal disease and oral cancer share common phenomenon: disruption of epithelial barrier integrity and inflammation. More specifically, the periodontal pathogen Fusobacterium nucleatum has been associated with both barrier disruption and with oral cancer. Still, it is unclear whether disruption of the oral epithelial barrier by Fusobacterium nucleatum is a mechanistic promoter of pro-tumorigenic oral transformation, or just a consequence of it. To answer this question, we interrogate here a cellular complex that is essential for epithelial barrier integrity, called the adherens junction. More specifically, we have found that the adherens junction component PLEKHA7, which is an E-cadherin and p120 binding partner, recruits core components of the RNA interference machinery including the microprocessor, the RNA induced silencing complex, and sets of mRNAs and miRNAs to maintain epithelial homeostasis. PLEKHA7 depletion from cultured epithelial cells results in mis- localization of the RNAi machinery, decreased miRNA silencing activity, increased oncogene expression, and pro-tumorigenic cell transformation. However, such disruption of the RNAi interference machinery, particularly associated with barrier integrity, has not been studied to date in oral cancer, exposing a significant gap in knowledge. We hypothesize that PLEKHA7 recruits and regulates the RNAi machinery at oral epithelial adherens junctions to maintain homeostasis, and that this mechanism is disrupted in the presence of oral pathogens, which in turn promote expression of pro-tumorigenic and inflammatory markers. We will test our hypothesis through two Specific Aims: 1) determine whether oral epithelial adherens junctions recruit the RNAi machinery to regulate function and levels of miRNAs and of their target mRNAs, using two dimensional and three-dimensional epithelial cell models and by examining tissues from patients; 2) determine if the oral periodontal pathogen Fusobacterium nucleatum promotes upregulation of oncogenic and pro-inflammatory markers through miRNA dysregulation, using co-cultures with Fusobacterium nucleatum, immunofluorescence, protein, and RNA analyses. This project is significant, since it will identify a novel molecular mechanism of epithelial pro-tumorigenic transformation in the oral mucosa, advancing our understanding of the disease. The study is innovative, as it links pathogens, oral epithelial barrier function, and localized RNAi and miRNA regulation with oral cancer. We anticipate that the study will be impactful and contribute to future development of effective biomarkers oral cancer, which are critically needed for timely and successful intervention, as well as to the potential development of RNA-based therapeutics. Altogether, this fellowship will provide foundational training in oral epithelial biology to foster the development of the DMD/PhD trainee into a unique and critically needed academic dental scientist.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT/PROJECT SUMMARY Ischemic heart disease presents a substantial burden of disease, partially due to cardiomyocytes’ low turnover rate. Neonatal mice exhibit a transient cardiac regenerative capacity and serve as a valuable model to study cardiac regeneration. Many in the field theorize that the driver of the regenerative to non-regenerative shift is the reactive oxygen species (ROS) increase that occurs after birth as pups shift from a low to a relatively high- oxygen environment. These ROS are thought to lead to DNA damage, resulting in cell cycle arrest. Although it is well established that high levels of ROS can drive pathology in a variety of tissue types, ROS serve as critical signaling molecules in a wide range of biological processes. One critical developmental process that occurs during the early neonatal period is the metabolic shift to fatty acid oxidation, which is a hallmark of cardiomyocyte maturation. The primary goal of this F31 research proposal is to determine the necessity of neonatal ROS increase in establishing oxidative tolerance in cardiomyocytes prior to ischemic injury (Aim 1) and in driving a metabolic shift to fatty acid oxidation that is characteristic of mature cardiomyocytes (Aim 2). This study includes an AAV9- delivered constitutively active Nrf2 as well as a high-dose antioxidant model to suppress ROS during the early neonatal period. Preliminary RNA sequencing data showed a decrease in genes associated with fatty acid oxidation in ROS-depleted cardiomyocytes compared to controls, suggesting that ROS are required for cardiomyocyte metabolic maturation. To further study the impact of abnormal metabolic shifts on cardiomyocyte regeneration, a Cd36 knockout and AAV9-delivered overexpression model will be utilized in combination with human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (iCMs) studies to manipulate cell metabolism. The proposed study focuses on mitigation of neonatal development ROS to understand their role in cardiomyocyte maturation and establishing oxidative tolerance, in contrast to studies that have aimed to lower injury-related ROS. While cardiac pathology has mainly been attributed to oxidative stress, recent findings suggest that reductive stress may also have pathological implications. Elucidating the temporal regulation of ROS signaling and its influence on cardiomyocyte injury response and metabolism will help inform future therapeutics for the treatment of heart failure.

NIH Research Projects · FY 2026 · 2024-01

Abstract Cardiomyocyte mitochondria generate ATP that fuels contraction and normal or reparative cardiomyocyte growth. The preferred metabolic substrates of cardiomyocyte mitochondria evolve during cardiac development from a fetal preference for carbohydrates to the normal adult preference for fatty acids, with reversion to fetal-like utilization of carbohydrates in adult cardiomyopathy. Much as gasoline and electric versions of the same automobile are not interconvertible by software “reprogramming”, we discovered that myocardial metabolic transitions require mitophagic elimination and biogenic replacement of carbohydrate-processing by fatty acid-processing mitochondria. We identified mitofusin (MFN) 2, which in other tissues is a mitochondrial fusion protein, as the key nodal regulator of mitophagic mitochondrial replacement in perinatal myocardial mitochondria, i.e. the hub of a mitochondrial dynamics-mitophagy interactome. However, our understanding of specific mechanisms that direct cardiac substrate utilization in adult hearts is incomplete, and forced biogenic production of cardiomyocyte mitochondria has not proven therapeutic in experimental models of heart disease. Although there is much work to be done before our findings can be translated into effective treatments for human heart disease, our research over the past several years has engendered a solid foundation for this goal. The conceptual breakthrough for this research was our discovery that MFN2 orchestration of mitochondrial fusion and mitophagy is the consequence of different MFN2 protein pairing events directed by specific PINK1-kinase phosphorylation events. A consequence of this mechanism is that mitochondrial fusion (MFN-MFN pairing) and mitophagy (MFN-Parkin pairing) are mutually exclusive. The biophysical process linking MFN2 phosphorylation to differential protein pairing is MFN2 conformational switching from a closed state favoring mitophagy to an open state favoring mitochondrial fusion. Research products generated from this work include the first mitofusin activating small molecules and an expanding catalog of MFN2 mutants available in adenoviral vectors and knock-in mice. Translationally, our work is beginning to identify clinical applications for pharmacological mitofusin activation. Here, we propose to translate what we have learned in basic mechanistic studies of mitochondrial dynamics factors to a preclinical delineation of disease mechanisms and evaluation of potential therapeutic approaches. Accordingly, we propose two goals: A basic research goal to determine how MFN2 multifunctionality relates to differential protein-partnering evoked by phosphorylation-induced changes in MFN2 conformation that expose or hide specific MFN2 protein binding domains. We posit that the unusually broad spectrum of mutational MFN2 dysfunction reflects, at least in part, differences in MFN2 protein partnering.Our translational research goal will determine if tissue-specific disease phenotypes caused by different human MFN2 mutations accrue from distinct patterns of MFN2 dysfunction due to mutational perturbation of specific protein pairing events. We predict that impaired MFN2-Parkin mediated mitophagy preferentially affects cardiac myocytes, while impaired MFN2-Miro regulated mitochondrial motility preferentially affects neurons. In pursuing these goals we will employ new concepts and reagents that we developed to dissect molecular mechanisms that drive metabolic remodeling in normal and diseased hearts, and to develop translatable means of optimizing myocardial metabolism by fine-tuning mitochondrial quality and quantity via precision manipulations within the mitochondrial fusion/motility/mitophagy interactome.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT Advanced neovascular age-related macular degeneration (AMD) is characterized by choroidal neovascularization (CNV), in which neovessels originating from the choroid invade the macula. CNV causes permanent central blindness if left untreated and is the most sight-threatening pathology of AMD. The present application will investigate the efficacy and regulatory mechanisms for a new class of CNV therapeutics. We have identified in our preliminary data that ketone bodies suppress CNV and hypothesize that this effect is regulated by macrophage and/or microglia reprogramming, which results in protective inflammation and/or suppression of pathological inflammation. We will refute or validate this hypothesis in three Specific Aims. In Aim 1, we will determine the therapeutic effects of ketone metabolites and their potential synergistic effect with anti-VEGF therapies, the current standard of care for CNV. We will also determine whether the anti-angiogenic effects of ketone metabolites are regulated by direct effects on endothelial cells or ketone-induced immune cell phenotypic changes using co-culture models. In Aim 2, we will identify whether ketone bodies suppress CNV via microglia, macrophages recruited from the circulation, or both. In Aim 3, we will determine the role of the ketone body receptor, G Protein-coupled receptor 109a (Gpr109a) in phenotypic reprogramming of specific immune cell subpopulations.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT Retained Foreign Objects (RFOs) are one of the most well recognized and persistent patient safety problems, affecting between 3000 and 50,000 patients per year, with an 80% morbidity rate, The primary defense is the manual counting of items that enter and leave the surgical field (the “surgical count”), which can be unreliable, while technologies, policies and training still leave many potential opportunities for failure. We propose a Patient Safety Learning Lab that will explore the causes of and solutions to RFOs using a systems engineering approach. Our initial systems models will drive improved understanding of factors that influence successful and unsuccessful counting. Laboratory experiments in count grouping, including time pressures, distractors, and the role of technology will define and build an evidence for optimal count strategies. It will also be possible to identify multiple opportunities to enhance the perioperative count process prior to the commencement of the procedure; and the discovery of lost items in the case of a discrepant count. Finally, as RFO performance is affected by organizational work contexts, we will explore opportunities to enhance teamwork, and organizational learning processes post-event. This work will initiate the most fundamental reappraisal of the causes of and solutions to RFOs for more than 20 years.

NIH Research Projects · FY 2026 · 2023-09

PROJECT SUMMARY Maladaptive alcohol seeking, a hallmark of alcohol use disorder (AUD), is thought to be driven not only by increased function of neural circuitry that drives natural reward seeking, but also by loss of control of circuitry that serves to suppress behaviors incompatible with safety and survival. The posterior paraventricular thalamus, (pPVT), and its projections to the nucleus accumbens (NAc), provide feedforward inhibition onto dopamine receptor-expressing medium spiny neurons (MSNs) which are largely responsible for promoting reward- motivated behaviors. Stimuli and situations that naturally serve to limit maladaptive behaviors, such as behavioral threats, have been shown to activate these pPVT®NAc projection neurons and suppress behavior through the activation of downstream parvalbumin inhibitory interneurons (PV-INs). Our labs have shown that an acute stressor (TMT predator odor), quinine-adulteration of alcohol and optogenetic activation of pPVT®NAc circuitry reduces reward- and, of particular importance to this proposal, alcohol-seeking in non-dependent mice. Furthermore, we show that the ability of this circuit to provoke behavioral inhibition is lost after the induction of alcohol dependence. These behavioral adaptations parallel reduced synaptic efficacy at pPVT®NAc glutamatergic synapses onto downstream PV-INs in alcohol-dependent mice. Our data therefore suggest, for the first time, that pPVT®NAc projection neurons are responsible for the suppression of alcohol-seeking behavior but are dysregulated by chronic alcohol exposure. We formally test independent components of this hypothesis in three independent Aims. In Aim 1, using two-photon calcium imaging we will measure and longitudinally track neuronal ensemble dynamics in both pPVT®NAc projections and NAc PV-INs during alcohol self-administration, consumption, and tests of behavioral inhibition in non-dependent and dependent mice. Aim 2 will explore the effects of chronic alcohol exposure on intrinsic and synaptic adaptations in pPVT synaptic inputs to both PV-INs and MSNs in the NAc using slice electrophysiology. Lastly, in Aim 3 we will determine whether activation of pPVT®NAc inputs and/or activation of PV-INs in the NAc are sufficient to restore feedforward inhibition and control alcohol-seeking in non-dependent vs. dependent mice. This project will identify how activity in a principal – but understudied – reward circuit changes from the onset of alcohol use to dependence and will further determine how this activity influences the expression and suppression of alcohol seeking.

NIH Research Projects · FY 2026 · 2023-09

PROJECT SUMMARY Metabolism of alcohol (i.e., ethanol) to acetaldehyde within different brain regions and in different subcellular compartments, and how that metabolism changes with chronic ethanol use, is not well-understood. The long- term goal is to identify adaptive changes that occur in chronic ethanol abuse and to identify therapeutic strategies to prevent or reverse neurological damage from ethanol. The objective of this proposal is to determine subcellular regulation of ethanol metabolism in mitochondria and endoplasmic reticulum (ER) by CYP2E1 during acute and chronic ethanol use, and to determine the consequences of the differential targeting. The central hypothesis is that targeting of CYP2E1 to mitochondria will be increased during chronic alcohol use in some regions of the brain that are sensitive to ethanol-induced damage, and that high mitochondrial targeting will drive high mitochondrial acetaldehyde production and resulting mitochondrial dysfunction and oxidative stress. The rationale underlying this hypothesis is that CYP2E1 expression overall is increased in brain regions that are sensitive to ethanol, including the prefrontal cortex, hippocampus, and cerebellum, and these regions also develop mitochondrial dysfunction and oxidative stress during ethanol use. The central hypothesis will be tested by pursuing three specific aims: 1) Evaluate subcellular specificity of induction of CYP2E1 by acute and chronic ethanol in the brain; 2) Determine the role of mitochondria- and ER-targeted CYP2E1 in mitochondrial effects of chronic ethanol use; and 3) Measure contribution of mitochondria- and ER- localized CYP2E1 to ethanol-induced oxidative stress. We will pursue these aims using an innovative strategy of three complementary systems: mice, C. elegans, and cultured cells. In each system we have a null background lacking CYP2E1, a wild-type CYP2E1 gene targeted to both mitochondria and ER, an ER-targeted CYP2E1, and a mitochondrial-targeted CYP2E1. The proposed research is significant because it will elucidate how ethanol metabolism by CYP2E1 changes over a chronic ethanol use paradigm, and could reveal mitochondrial CYP2E1 as a liability for ethanol toxicity. It is also significant because it generates useful platforms for studying subcellular localization-dependent effects of CYP2E1. The work will develop foundational resources that will be used by other researchers. The proximate expected outcome of this work is an understanding of how CYP2E1 contributes to the effects of ethanol in the brain during chronic and acute binge drinking. The results will have an important positive impact immediately because they will establish better understanding of the relationship between ethanol metabolism in brain regions with toxicity, and in the long-term because they lay the groundwork for identifying therapeutic opportunities.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY Alcohol use disorder (AUD) is characterized by the progression from recreational drinking to uncontrollable and excessive consumption resulting in a myriad of social and neurobiological complications. The mechanisms underlying the dependence-induced escalation in drinking are not completely understood. However, a key brain region disrupted in individuals with AUD is the orbitofrontal cortex (OFC). Studies from the Woodward laboratory show that acute ethanol inhibits action potential firing of lateral orbitofrontal (lOFC) cortex pyramidal neurons. This occurs via an astrocyte-dependent process involving activation of astrocytic D1/D5 dopamine receptors and the release of glycine via reversal of the GlyT1 glycine transporter. Following chronic intermittent exposure (CIE) to alcohol, lOFC neurons become hyperexcitable and are tolerant to acute ethanol. However, the effects of CIE exposure on lOFC astrocytes and how this affects lOFC neuronal excitability are not completely understood. The overarching hypothesis of this proposal is that CIE exposure impairs lOFC astrocyte function that contributes to hyperexcitability of lOFC pyramidal neurons and the resulting dependence-induced escalation in drinking. This hypothesis will be tested with two complementary aims. Aim 1 will test the hypothesis that CIE-induced increases in lOFC neuronal excitability and loss of acute ethanol inhibition involves astrocytic calcium signaling. To test this, male and female C57BL/6J mice will receive an intra-OFC infusion of an astrocyte-selective AAV encoding either a plasma membrane calcium ATPase (PMCA) or a reporter construct (tdTomato). Following repeated cycles of CIE exposure, slice electrophysiology will be used to measure current-evoked spiking of lOFC neurons and the membrane potential of lOFC astrocytes. Training in viral infusion surgeries and astrocyte and neuron slice electrophysiology will be achieved under this aim. Aim 2 will test the hypothesis that expressing PMCA in lOFC astrocytes prevents the increases in drinking following CIE exposure. In the first study, male and female C57BL/6J mice expressing either PMCA or tdTomato localized in lOFC astrocytes will undergo baseline sessions of two-bottle choice ethanol drinking. Weekly sessions of CIE exposure will then be interleaved with test weeks of drinking. The second study will follow the same CIE paradigm with the absence of homecage drinking and will use mice expressing GCaMP6f in lOFC astrocytes and the red-shifted opsin ChrimsonR in lOFC neurons. Training in astrocyte fiber photometry and optogenetics will be achieved under this aim. The proposed research studies will be complemented by career development activities including manuscript preparation, data presentation, networking, and training in the responsible conduct of research. These studies will provide novel insight into the role of lOFC astrocytes in AUD and position me to pursue a productive career in alcohol research.

NIH Research Projects · FY 2025 · 2023-09

Abstract: Chronic pancreatitis (CP) is a debilitating disease characterized by irreversible morphological changes (fibrosis) and persistent inflammation in the pancreas. Abdominal pain is one of the predominant symptoms and presents in up to 90% of CP patients and is associated with the heavy use of opioids. Chronic pain can cause opioid dependence, anxiety, depression and reduced quality of life. Although opioids are effective for acute pain, they are not effective as a long-term treatment strategy. There is a critical need for more effective, safer, and non- addictive therapeutic options for people who suffer from chronic pain associated with CP or other diseases. The use of mesenchymal stromal cells (MSCs) in treating pain represents a promising novel intervention as increasing evidence demonstrates that MSC therapy can effectively target several injury pathways in a variety of fibroinflammatory diseases while reducing pain, something that most pharmacological interventions cannot accomplish. MSCs exert protective effects through the release of pro-mitotic, antiapoptotic, anti-inflammatory, and immunomodulatory soluble factors, to mitigate metabolomic and oxidative stress imbalance. Data from animal models and clinical trials support the outstanding and durable effects of MSC infusion in the suppression of chronic inflammation and neurological pain associated with various diseases. Our own animal study demonstrated that MSC infusion significantly reduces pain and improves fibrosis and pancreatic volume in mouse models of CP. However, whether the infusion of MSCs can be used as a novel approach to relieving chronic pain and improve pancreatic function in CP patients has not been tested and will be the focus of this study. Based on compelling data, our hypothesis is that treatment with MSCs reduces chronic pain and improves pancreatic fibrosis and function by their ability to target multiple injury pathways. The objective of this study is two folds: (i) prepare for the clinical trial (UG3 phase, year 1), and determine the safety and efficacy of MSC therapy in patients with painful CP (UH3 phase, year 2-5). Another critical part of the clinical trial phase will be to define the mechanisms by which MSCs relieve multiple injuries in the pancreas by measuring changes of peripheral blood mononuclear cells (PBMCs), especially monocyte subsets and serum proinflammatory cytokine profiles of treated patients, with the goal of identifying biochemical pathways and serum biomarkers of response to MSC therapy. This study may help develop novel non-addictive cellular therapy for chronic pancreatitis and pain.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT BRCA2, as part of the DNA repair pathway, is a key regulator in homology-directed repair (HDR) and fork protection mechanism, which ensures genome instability. For full activity, BRCA2 must be transported into the nucleus to repair DNA double-strand breaks (DSBs). In normal cells, loss of nuclear BRCA2 can lead to genome instability and cancer but, in tumor cells, BRCA2 cytosolic mislocalization can lead to sensitivity to targeted therapies. I identified a paradoxical relationship between BRCA2 nuclear import and treatment. Pathogenic missense mutations in the DNA binding domain of BRCA2 direct the protein to the cytosol which increase sensitivity to PARP inhibitors (PARPi) and platinum drugs. Therefore, although BRCA2 mislocalization might be a cause of cancer, keeping it out of the nucleus allows for much better treatment outcomes. This has opened an avenue of research that I am perfectly aligned to study. In this K22 proposal I will address the nuclear transport of BRCA2, how cancer-driver mutations lead to cytosolic mislocalization and how BRCA2 localization can be used as a diagnostic tool. To test this, I will define the molecular mechanism(s) regulating nuclear/cytoplasmic shuttling of BRCA2 pathogenic missense variants (Aim 1), determine the impact of BRCA2 cellular localization on HDR, fork protection and cytosolic processes (Aim 2) and exploit BRCA2 localization as a cancer diagnostic tool (Aim 3). My priority will be to focus on missense variants identified during my postdoctoral career to decipher the underlying molecular mechanism of nuclear/cytosolic BRCA2 trafficking. At the successful completion of this K22 proposal, I will reveal novel pathways and factors that ensure proper localization of BRCA2 and how pathogenic BRCA2 missense variants have altered localization and functionality. This knowledge will give us a better understanding of the pathogenicity of BRCA2 missense variants and how we can modulate the functionality of BRCA2. My career goal is to obtain an independent position at a leading institution where I will dissect the functionality of missense variants in DNA repair proteins and how their cellular localization is important for genome stability as a tool to predict cancer risk and to treat cancer patients. My successful transition will be supported by advancing my expertise in cell biology, biochemistry, mass spectrometry and human cellular models. I will use these acquired skills to define why and how certain pathogenic BRCA2 missense variants are mislocalized to the cytosol. Importantly, the protected time that this award provides me will allow me to elucidate the factors and pathways by which BRCA2 is transported from the cytosol to the nucleus, how this might be altered in BRCA2 missense variants and how this could be used for targeted therapies. Furthermore, the success of this project will be greatly enhanced by the outstanding advisors and collaborators that advise me through the K22 period. The receipt of this award will allow me to expand my research plan and establish myself as a principal investigator in the field of cancer biology.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT After a stroke, walking ability can be compromised, which can lead to reduced quality of life and decreased ability to perform activities of daily living. Post-stroke walking recovery is mediated by nervous system reorganization (e.g., neuroplasticity), however our understanding of these processes related to improvements in walking function are limited due to the neurophysiological complexity of walking itself. Additionally, current practices of assessing stroke- impacted neuroplasticity are heavily focused on the motor system. Of keen interest is the integration of sensory and motor systems (e.g., sensorimotor integration-SMI), which are necessary for initiating, sustaining, and coordinating walking while providing continuous feedback on body state and actions. Additionally, impaired sensation post-stroke can lead to lower extremity dysfunction, balance problems, and falls, highlighting the need to investigate the effects of stroke on SMI in relation to walking function and recovery. Given the importance of SMI in walking, the capacity for these sensorimotor networks to exibit plastic changes may be a crucial mediator for improvements in walking funciton, thus necessitating relevant methods of assaying sensorimotor plasticity. To address this gap in knowledge, in the F99 Phase (Aim 1) I will characterize lower extremity sensorimotor plasticity in individuals post-stroke. I will specifically target the connections between the primary sensory cortex (S1) and and the primary motor cortex (M1) using paired associative stimulation (PAS), a novel method of assaying sensorimotor plasticity. PAS is based on the Hebbian principle of associative plasticity, in that repetitive stimulation of pre- and post-synaptic neurons within S1 and M1 leads to increased synaptic efficacy, evidenced by rapid and long-lasting increases in transcranial magnetic stimulation-induced muscle responses, or motor evoked potential (MEP) amplitude. We intend to account for individual differences in SMI by adjusting the timing between stimuli, and measuring the time it takes the electrical stimulation to reach S1, via somatosensory evoked potentials. Changes in MEP amplitude following PAS are thought to reflect the induced associative plasticity between S1 and M1, and are shown to be reflective of upper extremity function, with limited work done in the lower extremities. Therefore, we hypothesize that individualized PAS protocols, will facilitate increases in MEP amplitude, and changes will be associated with clinical measures of walking function. In the K00 Phase (Aim 2), I will pursue advanced training in post-stroke sensory and motor reorganization via multi- modal techniques to understand walking and balance recovery. I will seek out postdoctoral mentorship that will allow me to build on my predoctoral work by acquiring advanced electrophysiological and neuroimaging skills. I will also focus on individualized assessments and the identification of mechanisms underlying the response to treatment between individuals over the course of rehabilitation. The F99/K00 will help facilitate my path to independence through research training and career development, enroute to my long-term goal of becoming an independent researcher at an academic medical center. The proposed work and career direction aim to inform lower extremity rehabilitation efforts by providing functional and structural neural correlates of recovery.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT The impact of heart transplantation (HTx) remains limited by donor shortages, with an estimated 250,000 adults who may benefit from transplant despite only 3,500 being performed each year in the US. Unfortunately, donor discard rates remain high at 70-80%, with substantial unexplained variability in donor evaluation and acceptance practices between centers. Recent data also demonstrate that higher risk recipients are being transplanted under the new 2018 allocation policy with worse post-transplant survival rates nationally. These trends collectively underscore current limitations in allocation policy and the ability for individual programs to assess donor quality and to pair suitable donors with appropriately selected recipients. The latter stems from a suboptimal process whereby clinicians have to make time-sensitive decisions relying solely upon experience and judgement without data-driven tools that can analyze numerous donor and recipient data and their complex interactions to provide rapid and accurate outcome projections. Existing risk models have failed to garner widespread utilization due to major limitations, including 1) narrow focus on only one of a set of relevant outcomes, 2) simplistic approach with only modest discriminatory capability (c-statistics <0.70), 3) failure to account for complex interactions between donor and recipient variables, and 4) use of only static, cross-sectional data. Our proposal seeks to advance the field by leveraging a novel, comprehensive dataset and machine learning (ML) to develop robust models that can maximize predictive performance for relevant outcomes and to better align a candidate's clinical trajectory and anticipated transplant outcome. These models will better account for complex interrelationships between donor and recipient variables, and will also account for dynamic changes in candidate and donor parameters. Optimized models will then be incorporated into a decision support system guided by key stakeholders. In addition, a previously developed artificial intelligence (AI) framework will be used to optimize heart allocation policy. We have these specific aims: 1) Establish the feasibility and usability of a stakeholder-guided, ML-derived decision support system for adult HTx; 2) Demonstrate the adaptability of a previously developed AI-based policy-optimization framework to heart allocation; and 3) Inform and evaluate the processes and outputs of Specific Aims 1 and 2 using stakeholder engagement and implementation science to refine and optimize working prototypes and promote the understanding, adoption, and use of data-driven decision support tools created for HTx. This work will optimize the allocation of scarce resources and ultimately improve outcomes of HTx.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Approximately 29.5 million people over the age of 12 in the US had alcohol use disorder (AUD) according to the 2021 National Survey on Drug Use and Health. Chronic alcohol use can lead to the emergence of cognitive deficits, particularly working memory, which persist during alcohol withdrawal and abstinence increasing the odds of relapse. Approved treatments for alcohol use disorder are not designed to treat alcohol-induced cognitive dysfunction, so understanding potential mechanisms is necessary to develop new treatments. The nucleus reuniens of the thalamus (RE) is a ventral midline thalamic nucleus that plays a key role in cognitive function, such as spatial working memory, attention, and behavioral flexibility. The RE is also bidirectionally connected to the medial prefrontal cortex (mPFC) and the hippocampus, which have been highly studied in AUD and cognition. Despite its interconnectedness, the RE is understudied in the alcohol field. In my preliminary studies, I induced alcohol dependence using chronic intermittent ethanol (CIE) vapor exposure (4 cycles) in male and female C57BL/6J mice. Following CIE, mice were either tested on a T-maze delayed alternation spatial working memory task or sacrificed for whole cell patch clamp electrophysiology to measure intrinsic excitability in the RE. Mice used in the T-maze task were trained prior to CIE and tested following 2 and 4 cycles, to track cognitive decline across the development of dependence. Both male and female mice exhibited significant deficits in performance after CIE. Despite this similar behavioral phenotype, CIE had divergent effects on intrinsic excitability, causing an increase in firing in females and a decrease in males. I also found that firing in the RE in the absence of alcohol exposure appeared to be sex-dependent, where males have significantly greater excitability than females. These results indicated that the RE could be an exciting new target for the study of chronic alcohol- induced cognitive deficits. The overall hypothesis of this research proposal is that chronic alcohol exposure causes sex-dependent functional adaptations to the RE and that regulating RE activation will improve cognitive performance. To achieve this, the proposal uses a multi-technical approach to study the RE in vivo and in slice physiology. Aim 1 of this proposal will determine if chronic alcohol exposure alters the function of the RE using fiber photometry during performance on a spatial working memory task and whether normalizing aberrant activity using optogenetics will improve cognitive performance in CIE-exposed mice. Aim 2 of this proposal will characterize the effect of CIE exposure on the physiology of RE neurons involved in mPFC and hippocampal circuitry by recording intrinsic excitability and synaptic transmission from neurons that project to the mPFC, the hippocampus, or both. The results of these experiments will serve as landmark studies for the role of the RE in alcohol-induced cognitive dysfunction and allow us to further understand the underlying mechanisms of cognitive deficits following chronic alcohol.

NIH Research Projects · FY 2024 · 2023-09

The deep-brain imaging approach (microendoscopy) is a high throughput method that identifies the temporal and spatial pattern of activity in specific phenotypes of neurons. We have hypothesized that activity in small networks can identify normal versus diseased brains. To test this hypothesis, we will use the two-wavelength miniscope to image activity of a subset of neurons in 5xFAD mice, a validated animal model of Alzheimer’s disease (normal versus the homozygous littermates). We have created promoter- driven calcium sensor that drives the red-shifted calcium indicator, jRGECO, into orexin-arousal neurons. A separate calcium sensor (GCaMP6m) in GABA sleep neurons will be used as comparison. The advantage of imaging neurons with two different wavelengths is that it allows for comparison of arousal versus sleep neurons as the brain transitions between wake and sleep. We will image activity of neurons that contain jRGECO and GCaMP during normal wake-sleep bouts and after 6h sleep loss. We hypothesize that there is an abnormal temporal pattern of fluorescence during the transition from wake to sleep in the brains of 5xFAD mice, and it worsens with progression of disease. This may explain the cause of the disrupted sleep-wake patterns in Alzheimer’s disease. We hypothesize that abnormal activity in deep brain circuits regulating sleep is a harbinger of disease. The overall impact of this small budget project is that it will identify activity of two different juxtapositioned neurons during wake, NREM and REM sleep in wildtype versus a disease model, which will aid in understanding how small clusters of neurons behave as the waking brain falls asleep.

NIH Research Projects · FY 2025 · 2023-09

Abstract The majority of smokers try to quit each year, and the majority of quit attempts fail, even when the most effective FDA-approved pharmacotherapies are used. Non-combustible tobacco products emit fewer harmful chemicals than cigarettes, and thus for smokers who cannot quit smoking, switching completely to a less harmful product is likely to improve their health. For smokers who have failed to quit with traditional methods, trying to switch to a less harmful product may be more likely to help them stop smoking than trying to quit using tobacco altogether repeatedly with pharmacotherapy. The proposed trial evaluates the potential of non-cigarette tobacco products to serve as harm reduction tools for current smokers who have already tried, and failed, to quit with traditional methods. Current smokers who failed to quit with FDA-approved pharmacotherapy within the past year (N=225) will be randomly assigned to either 1) switch completely to a non-cigarette tobacco product (Switch Group, n=150), or 2) try to quit again using pharmacotherapy (Meds Group, n=75). Participants will choose among a limited menu of products or medications they would like to receive (maximizing external validity) and select a Target Switch / Quit Date on which they will stop smoking. Participants in the Switch Group will choose between two non-cigarette products that deliver sufficient nicotine and are appealing to adult smokers: e-cigarettes, the most commonly used non-cigarette tobacco product, or heated tobacco products, a novel tobacco product recently approved for sale by FDA as modified risk exposure compared to cigarettes. Both product classes, while not safe, emit fewer harmful chemicals compared to cigarettes. Participants in the Meds Group will choose between the two most effective pharmacotherapies available: varenicline or combination transdermal nicotine replacement therapy and short-acting nicotine lozenge. Participants in the Switch or Meds groups will receive a 9-week supply; broken down as: 1-week to use ad libitum while continuing to smoke, and 8-weeks to use as instructed following a Target Switch or Quit Date. The primary outcome is biochemically-confirmed abstinence from cigarette smoking at the 6-month follow-up, and secondary outcomes include smoking reduction > 50%, longest quit attempt duration, biomarkers of nicotine, smoke, and toxicant exposure, and changes in respiratory symptoms. The proposed trial addresses a highly significant research question using a rigorous design and is supported by a strong investigative team. The trial is in line with priorities within NOT-OD-22-023 including “Harm reduction studies that involve switching from combustible cigarettes to ENDS [or heated tobacco products].”

NIH Research Projects · FY 2025 · 2023-09

Abstract – Mentorship in digestive disease research areas is lacking. The reasons for this are complex, and have been attributed to a variety of factors. However, we have recognized that a major issue is the lack of effective mentorship. The overall objective of the Medical University of South Carolina (MUSC) contribution to the Helping to Accelerate Research Potential (HARP) Program in NIDDK related disease areas is to provide ready access to quality mentoring and training - thereby enabling outstanding research in NIDDK related disease areas. Our vision is to establish the HARP program as a sustainable resource for NIDDK related disease areas throughout the U.S. Through the HARP Program in Digestive and Liver Disease, we have hypothesized that a mentoring network to supplement that which is present in the mentee's university will provide additional critical and valuable mentorship. Therefore, we propose to implement innovative and transformative researcher mentorship and training to ensure development and sustainability of the digestive and liver disease workforce. We have proposed the 3 Specific Aims as follows: Aim 1 - to establish durable mentoring relationships and to effectively mentor a cadre of talented early career scientists to become independently funded in NIDDK related research areas through an integrated and dynamic mentorship program. Aim 2 - Increase scholars' scientific knowledge, professional skills, motivation, and credentials to pursue independent careers in NIDDK-related research areas via a rigorous educational and didactic program. Aim 3 – to promote the long-term viability and success of the HARP Program in NIDDK related research areas through rigorous evaluation, ongoing improvement strategies, and mission alignment with other HARP mentors and centers. The MUSC HARP program will be supported not only by MUSC's CTSA, but also by its rich and experienced institutional training programs. In summary, through expansion of mentorship of junior investigators and promotion of interdisciplinary exchange among mentors, Scholar's, and NIDDK leadership, the MUSC HARP Program will substantially augment thematic NIDDK related disease research on a national level. The founding efforts proposed here and in collaboration with the entire HARP group are expected to establish an effective, high quality, sustainable, and multidisciplinary career development program.

NIH Research Projects · FY 2025 · 2023-09

Animal and epidemiological studies show that prenatal experiences (e.g., elevated environmental pollutants, pregnancy complications, preterm birth, stress) support the ‘developmental origins of health and disease’ model in relating adverse maternal experiences to compromised fetal and child development. Hypotheses suggest that the developmental window of plasticity for obesity programming initiates in utero and extends through the first two years of life, underscoring the large impact that gestational and early life exposures and corresponding targeted interventions may have on lifetime obesity risk. Current obesity interventions may be ineffective because they target children or adults past the critical age when developmental programming occurs. The ECHO pregnancy cohort has the potential to address this critical gap in knowledge. Moreover, because previous research has often been conducted in urban areas, rural communities are often understudied. Hence, as a rural state with poor birth outcomes, where few studies have been conducted, our contribution to ECHO will have a major impact. Therefore, we will recruit over 500 pregnant women and their resulting offspring into ECHO from the Medical University of South Carolina (MUSC) and implement the ECHO-wide protocol including specialized components on Physical & Chemical exposures and child Obesity outcomes. MUSC has a large obstetrical/delivery population (i.e., over 3,000 annually) that facilitates meeting our recruitment goals. Our aims pertaining to ECHO-wide analyses are (1) to determine the joint impact of prenatal exposure to chemical mixtures and early gestational and life exposures (i.e., hypertensive disorders of pregnancy, preterm birth) on child obesity and metabolic health; (2) to identify differences in prenatal chemical exposures and their sources (dietary choices/opportunities, use of consumer goods and personal care products, occupation, air/water) among ECHO cohort participants; and (3) to determine the association of mixtures of pre-conceptual and peri-conceptual chemical exposures measured in mothers and fathers with hypertensive disorders of pregnancy, preterm birth, and child obesity. In summary, our proposed aims are innovative because they address: (1) the impact of joint exposure to chemical and non-chemical stressors; (2) potential rural-urban differences in sources of chemical exposure and resulting complex chemical mixtures; and (3) determining critical windows of exposure to complex chemical mixtures.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY/ABSTRACT Age-related hearing loss (ARHL) is a highly prevalent chronic condition and is associated with poorer psychosocial well-being, communication, and cognitive function. Evidence that hearing aid (HA) use mitigates negative consequences of ARHL is inconclusive because, in part, longitudinal population-based studies of hearing with sufficient follow-up times are rare and few have considered the extent to which earlier intervention with HA improves long-term HA outcomes and reduces the wide-ranging consequences of ARHL. The long- term objective is to reduce impacts of ARHL by maximizing overall HA benefit. The specific objective is to holistically assess benefits of HA use on psychosocial well-being, communication, and cognitive outcomes that capture general and hearing-specific health and determine the extent to which earlier intervention for ARHL is a determinant of HA benefit. To achieve this objective, two specific aims are proposed. Aim 1 will use longitudinal data from the Medical University of South Carolina (MUSC) Longitudinal Cohort Study of ARHL to determine benefits of HA use and assess contributions of early intervention and test hypotheses that a) use of HA will be associated with better general health aspects of psychosocial well-being, cognition, and communication, and b) earlier intervention with HA will be associated with greater HA benefit as determined by these outcomes. Aim 2 will use cross-temporal data to determine benefits of HA use and assess contributions of early intervention using historic and newly collected data from participants in the MUSC Longitudinal Cohort Study of ARHL to test hypotheses that a) use of HA will be associated with better hearing-specific psychosocial well-being and communication outcomes, and b) earlier intervention with HA will be associated with greater HA benefit as determined by these outcomes. Results from this project will positively impact public health research and clinical practice. Specifically, results can inform future epidemiological research and/or clinical trials aiming to determine effects of early intervention on HA benefit, optimal timing of HA interventions, and the appropriate age to begin screening for ARHL. This project and line of research directly addresses the National Academies of Sciences, Engineering, and Medicine (2016) recommendation to improve prospective population-based evidence on impacts of ARHL and its treatment and aligns with NIDCD’s strategic plan on secondary and tertiary prevention and health promotion for individuals with ARHL. That is, this proposal aims to determine importance of early intervention for ARHL (secondary prevention) and benefits of HA use (tertiary prevention) on general and hearing-related measures of psychosocial well-being, communication, and cognition. This project and training plan will provide rigorous mentored training in new methods focused on epidemiological and public health research in audiology, grantsmanship, teaching, and career/professional development, and will support the applicant’s long-term goals of being an independent, NIH-funded clinician-researcher.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT The co-use of nicotine/tobacco products and cannabis among young adults is prevalent and varies widely in terms of patterns and products of use. Nicotine-cannabis co-use among this age group may adversely affect treatment and other clinical outcomes, yet little is known regarding the underlying relationship between nicotine and cannabis and the resulting treatment implications, particularly during an attempt to quit or reduce use of one or both substances. Prior literature on the treatment implications of co-use, including results from our group, suggest that cannabis serves as an obstacle to nicotine cessation for a sub-set who use both products, though the literature is mixed. Past studies also suffer from important limitations, resulting in critical gaps in our understanding, with only 2 secondary analyses published from youth tobacco trials on the impact of co-use. To date, there are no prospective studies that have examined the treatment implications of nicotine-cannabis co- use or the underlying relationship between substances among young adults when engaged in a quit attempt. [[[Therefore, the overall goal of this project is to characterize and evaluate the underlying relationship between nicotine/tobacco and cannabis and its impact on nicotine cessation through a behavioral economics framework. This study will determine how impactful cannabis co-use may be on nicotine cessation and which individuals who co-use experience greater difficulty achieving cessation. To accomplish this goal, we propose a completely remote, prospective, 12-week nicotine cessation trial among young adults (ages 18-25; N=350) from across the United States who use nicotine (vaping, cigarettes or both) and cannabis products regularly. We will leverage our ongoing collaboration with DynamiCare Health to administer remote contingency management (CM) as the nicotine/tobacco treatment intervention, while cannabis use will not be addressed. Biochemical verification (through oral fluid samples) and self-reports (mobile daily diaries) of substance use will be collected. The aims of this proposed study are to: 1) evaluate the impact of behavioral economically derived measures of substance substitutability on end of treatment nicotine abstinence (Aim 1); 2) determine if treatment-induced nicotine abstinence, reduction, and/or withdrawal (a) is associated with co-occurring changes in cannabis demand and use and (b) if substance substitutability modifies this relationship (Aim 2); and 3) assess the reciprocal prospective relationship between patterns of nicotine and cannabis use during nicotine treatment (Aim 3). Evaluating nicotine-cannabis co-use as part of a prospective treatment study through a behavioral economics framework has never been conducted and will be important to informing the literature regarding individual co- use pattern differences, compensatory cannabis use, and treatment success.]]] Nicotine cessation treatment in young adults is a growing body of work and the role of cannabis co-use, and results from the proposed study, will inform clinical care and guide tailored treatment recommendations to promote successful and sustained abstinence.

NIH Research Projects · FY 2025 · 2023-09

Project Summary We used a human iPSC-derived hepatocyte platform to identify a family of structurally related small molecules that can reduce the production of APOB by the liver. These small molecules are highly effective, do not cause abnormal lipid accumulation, and have a chemical structure that is distinct from any known cholesterol lowering drug. Such compounds have the potential to treat hypercholesterolemia and steatosis. In the current application we propose to define the molecular mechanisms through which the compounds act. In preliminary data we show that the compounds inhibit human carboxylesterase 1 (CES1). In the three proposed aims we will i) identify the binding characteristics of the compounds to CES1 and define their specificity using crystallography and biochemical assays, ii) use iPSC-hepatocytes to definitively establish the role of CES1 in mediating (V)LDL-cholesterol production by the compounds, and iii) determine the efficacy of the compounds in lowering cholesterol and steatosis humanized mouse models.

NIH Research Projects · FY 2024 · 2023-08

Major barriers to chronic graft-versus-host disease (cGVHD) research are the inability to predict the likelihood of response to cGVHD therapy and subsequent patient survival. This absence of predictive biomarkers is partly due to the complex pathology of cGVHD, which involves both soluble and cellular factors but mostly due to the paucity, so far, of samples collected when specific cGVHD treatments are initiated, particularly as novel and more targeted treatments for cGVHD are now available. Thus, major questions remaining in the field are: Can we target more than one cGVHD pathway with a single drug? Can we detect biomarkers to predict future resistance to treatment using already validated cGVHD biomarkers? Can we discover more specific markers through a high throughput proteomics pipeline? Can we validate and utilize these predictive biomarkers to provide strong support for FDA approval of these biomarkers? For this project, we will use samples (plasma and PBMCs) collected during Dr. Pavletic trial testing Pacritanib, a multi-kinase inhibitor with specificity for JAK2 and IRAK1, in patients with steroid-refractory/steroid-dependent (SR/D) cGVHD to analyze proteomic signatures associated with prediction of cGVHD therapy nonresponse, and with prognosis of nonrelapse mortality (NRM). Proposed markers are based on previous studies and will include other novel or hypothesized factors. We will test the hypothesis that plasma proteomic panels measured prior to the start of cGVHD treatment with pacritinib (PAC) will stratify HCT patients for prediction of resistance, and NRM at 1 year. We will determine the thresholds of different biomarkers and panels that provide best sensitivity and specificity. Our overarching hypothesis addresses gaps remaining by addressing two specific aims (SA): SA1: Are five previously identified plasma cGVHD biomarkers [chemokine (C-X-C motif) ligand 9 (CXCL9), CXCL10, matrix metalloproteinase 3 (MMP3), Dickkopf-related protein 3 (DKK3), and Stimulation 2 (ST2; the interleukin (IL)-33 receptor)] predictive of resistance to PAC therapy? We will measure them using ELISA from fresh blood samples collected during this award: ~35 samples pre-treatment and 3- and 6-months post-treatment. SA2: Can additional plasma biomarkers that are key biologic drivers of cGVHD resistance be discovered through our proteomics pipeline comparing responders vs. non responders? We will address this question using our well- established proteomic workflow that can identify and quantify more than 2000 proteins. Upon completion, these studies will result in biomarker panels that may facilitate prediction of response and resistance to therapy and identify candidates for new therapeutic approaches.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY/ABSTRACT Compared to either disorder alone, co-occurring alcohol use disorder (AUD) and posttraumatic stress disorder (PTSD) has a more complex clinical and treatment course that imposes significant health burden on veterans. Identifying novel mechanisms to improve integrated AUD/PTSD treatment outcomes is a public health priority that aligns with NIAAA’s strategic goal to advance treatment of alcohol-related conditions through the refinement of behavioral and pharmacological treatments. Self-compassion is a cognitive skill that may facilitate more effective integrated AUD/PTSD treatment. Self-compassion is proposed to function by tapping into an innate care-giving system that could interact with oxytocin’s prosocial effects such as feelings of safety and trust. The current study will leverage an ongoing NIAAA-sponsored clinical trial that is examining the efficacy of oxytocin compared to placebo to improve outcomes in an integrated behavioral treatment among veterans with AUD/PTSD. The primary goal of the proposed study is to examine the bivariate longitudinal associations between self-compassion and treatment outcomes. The secondary goal is to examine whether veterans higher in self- compassion and administered oxytocin will outperform veterans in the placebo condition. Finally, we will utilize ecological momentary assessment (EMA) to capture the associations between self-compassion, alcohol consumption and problem severity, and PTSD symptoms in a real-world context. We hypothesize that self- compassion will enable more adaptive responding to AUD and PTSD symptoms (e.g., managing craving, lapses, maladaptive thinking patterns) thereby promoting a more efficient treatment course and reducing susceptibility to risky alcohol use or relapse. This is an important and feasible project that will provide novel insights into the role of self-compassion in AUD recovery among veterans with co-occurring PTSD. This project will also provide critical training experiences necessary to facilitate the candidate’s long-term goal to be an independent alcohol researcher. Under the proposed award, the candidate will have the opportunity to 1) master the extant literature on behavioral and pharmacological interventions for AUD/PTSD; 2) acquire new skills in the design and implementation of clinical trials; 3) hone skills in naturalistic assessment methods such as EMA; 4) advance longitudinal data analytic capabilities; and 5) increase scholarly productivity through grant and manuscript development. She will accomplish these deliberately selected training goals by leading the proposed research project, engaging in a comprehensive mentorship plan with highly skilled sponsors, and participating in a carefully selected program of didactic training opportunities. The training will be carried out at the Medical University of South Carolina (MUSC) in the Addiction Sciences Division. MUSC is a renowned academic medical center known for its innovative and excellence in alcohol research. Thus, the candidate will have access to an institution and sponsors with a strong history and commitment to clinical alcohol research.