Medical University Of South Carolina

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Nonsense or missense mutations of the CHAMP1 gene, which encodes for a zinc-finger protein involved in the maintenance of kinetochore-microtubule attachment during mitosis and regulation of chromosomal segregation, causes a neurodevelopmental syndrome characterized by intellectual disability (ID), ASD-like behaviors, microcephaly, hypotonia, and dysmorphic features. CHAMP1 interacts with POGZ, which encodes for a gene that causes the White-Sutton syndrome, and it is known that the interaction of these two proteins is lost in the presence of mutations. To date, 36 disease rare variants in CHAMP1 have been reported across studies comprising nonsense, missense, and frameshift mutations. Recently, a Champ1+/- mouse model showed relatively mild learning and memory impairment and depression-like behaviors and delayed neuronal differentiation. Despite the mouse data, there is yet no study that focused on the human brain leaving the role of CHAMP1 in human brain development totally unexplored. As such, our central hypothesis is that CHAMP1 loss of function alter neurogenesis and neuronal migration in human brain resulting in dysfunction of cortical local circuits. We propose a series of complementary approaches, including high-resolution imaging, single-cell multiomics, and electrophysiology tools to explore CHAMP1 loss of function in the following specific aims: Specific Aim 1. Determine the influence of CHAMP1 loss of function in the development of forebrain cortical organoids. Specific Aim 2. Profile single cell level transcriptomic and epigenomic landscapes of CHAMP1 loss of function organoids at different developmental stages. Specific Aim 3. Determine the effect of CHAMP1 loss of function in neuronal activity and local network.

NIH Research Projects · FY 2025 · 2024-09

Abstract Neonatal Intraventricular hemorrhage (IVH) originates from the underdeveloped germinal matrix, a site of cell rapid cell division adjacent to the lateral ventricles of the brain. IVH leads to brain injury and life-long neurological disability. Within the same time frame that neonatal IVH occurs, the brain is rapidly producing the cells needed for myelination. Oligodendrocytes, the myelin-forming cells of the brain, are derived from oligodendrocyte progenitor cells (OPCs). OPCs are fragile cells - exquisitely sensitive to many factors present across multiple neurological diseases such as excitotoxicity, inflammatory cytokines, and oxidative stress. Across a wide spectrum of neurological diseases, neuroinflammation disturbs OPC development. IVH both causes inflammation and leads to white matter pathology. Microglia, the endogenous macrophages of the brain, are the main effectors of neonatal neuroinflammation and are implicated in many forms of neonatal brain injury. However, microglia also produce insulin-like growth factor-1 (IGF-1), which is required for normal OPC development. IGF-1 is decreased by other forms of neonatal brain injury but has not been studied in IVH. Our preliminary data indicates that microglia increase production of inflammatory cytokines and reduce IGF-1 production after exposure to hemoglobin. Both of these effects could impair OPC development. Supplementing IGF-1 after IVH may be a powerful therapy because IGF-1 not only supports OPC health, but also modulates inflammation and reduces production of cytokines that are harmful to OPCs. These dual roles of IGF-1: supporting OPC development and altering the phenotype of immune cells (including microglia), makes it an excellent candidate for treating IVH in which both inflammation and white matter pathology occur. This project is designed to better understand how microglia respond to IVH, with a focus on how hemoglobin and IGF-1 affect inflammation and OPC development. It is also designed to trace OPC fate after IVH and determine if myelination failure is due to OPC loss versus maturation failure. We will trial IGF-1 as a therapy for IVH, and test its ability to modulate microglial activation and improve myelination.

NIH Research Projects · FY 2025 · 2024-09

Abstract – MUSC Gastroenterology (GI) and Hepatology Medical Student Training Program There is currently a crisis in physician led research in academic medicine. The 2021 NIDDK Strategic Plan for Research specifically recommended that there be an effort to “Bolster workforce development and training to increase and diversify the pipeline of clinical investigators.” The report further emphasized the critical importance of “Addressing the workforce pipeline at earlier career stages”. We believe that Medical Students are at the peak of their formative transition, and thus engaging them in GI and liver disease research is highly responsive to the NIDDK call for action. Training scientists in biomedical research requires specific scientific expertise, including a firm understanding of topic matter and the ability to generate novel and relevant research hypotheses. Through our expertise and experience, we are in an ideal position to provide the training necessary for Student's long term success. Further, current evidence suggests that mentorship is one of the most critical requirements for successful engagement of trainees at all levels, particularly in formative years of training, such in medical school. Multiple scientific organizations have called for quality mentoring opportunities for trainees. A needs assessment of MUSC Medical Students has identified 2 major areas of need related to GI and liver disease research on the Medical University of South Carolina (MUSC) campus. First, access to skilled and successful researchers is required. Secondly, a structure around training and development of projects, embedded in a matrix of quality mentorship, is required. Therefore, the overall objective of the MUSC Gastroenterology (GI) and Hepatology Medical Student (MUSC-GIHMS) Training Program is to enable Medical Students to participate in high quality research during their medical school training. Our vision is to provide access to quality research projects, and while doing so, engage, inspire, and excite a diverse group of medical Students in the field of GI and hepatology. In an effort to be highly responsive to the NIDDK Funding Opportunity announcement, PAR-21-034, the Specific Aims of this application propose to (1) provide both a focused research experience during the spring semester of second year for MUSC Medical Students, (2) to provide advanced curricular course work in digestive and liver disease research for Students, and (3) to continuously evaluate and improve the program so that it is sustainable over time. The current proposal will provide Students with an ideal setting to participate in GI and liver disease research and ultimately through recruitment of these committed and diverse Medical Students, will expand the pool of researchers engaged in GI and liver disease research.

NIH Research Projects · FY 2025 · 2024-09

PROJECT ABSTRACT The prevalence of cigarette smoking among individuals with substance use disorders (SUD) is nearly three times higher than the prevalence of smoking among the general population. Individuals with SUD are more likely to die from smoking-related illness than from complications associated with substance use. In addition to the direct deleterious health effects of tobacco, continued smoking during SUD treatment is associated with worse substance-related outcomes and greater odds of relapse. However, few SUD programs offer tobacco cessation services and even when available, quit rates for individuals with SUD remain lower than the general population. Tobacco cessation interventions that have been shown to be effective in general SUD treatment settings have been multicomponent interventions that are time- and resource-intensive and expensive to implement. Thus, there is a strong need to implement low-intensity, appealing interventions that can reduce the harms from combustible tobacco among individuals in SUD treatment. Tobacco harm reduction strategies to reduce tobacco- related morbidity and mortality among an SUD treatment population are an appealing and pragmatic approach that may lead to greater uptake among clients, as well as program staff and leadership and may provide a viable alternative to better serve clients (when deemed appropriate, but not as a replacement to cessation). One harm reduction strategy is to ask smokers to switch completely to a non-combusted, electronic nicotine delivery system (ENDS). A handful of rigorous RCTs have now shown that switching to ENDS has the potential to promote abstinence from cigarettes better than traditional forms of pharmacotherapy in a general population of those who smoke and are motivated to quit. However, switching to ENDS as a harm reduction strategy has yet to be compared to pharmacotherapy among individuals with SUD (efficacy) and evaluated for acceptability and appropriateness among clients, program staff and leadership (implementation). Further, smoking cessation during SUD treatment has been shown to improve SUD outcomes and therefore it is critical to evaluate the impact of tobacco harm reduction interventions on SUD outcomes and engagement. This study proposes a Hybrid Type 1 effectiveness-implementation randomized controlled trial to evaluate the impact of switching to ENDS compared to quitting tobacco with an FDA-approved pharmacotherapy for smoking cessation (nicotine replacement therapy [NRT]). This trial will enroll 240 clients across 3 South Carolina outpatient SUD programs. Participants will be randomized 1:1 to either switch to an ENDS product or to quit smoking with NRT. We will 1) assess whether switching to ENDS better promotes biochemically-verified point prevalence abstinence from cigarettes at the 12-week end of treatment visit compared to NRT (Aim 1), 2) evaluate patient, provider, and organizational-level implementation outcomes using mixed methods according to the Proctor framework (Aim 2), and 3) assess SUD treatment outcomes and engagement between groups to test for non-inferiority (Aim 3).

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY/ABSTRACT Intimate partner violence (IPV), posttraumatic stress disorder (PTSD), and opioid use disorder (OUD) are highly co-occurring conditions. Up to 94% of individuals using opioids report current or past IPV, and 64% of individuals with IPV meet diagnostic criteria for PTSD. There are increasing calls for all people to be screened for IPV across healthcare settings. However, to date, there are no well-established evidence-based screening, assessment, and referral programs for IPV or its consequences, such as PTSD, in opioid treatment programs (OTPs). Validated assessments and evidenced-based care programs—such as violence shelters and PTSD treatments—exist; however, current data show valid assessment tools may not be routinely implemented in OTP settings and that treatment providers in these settings may not feel confident in their ability to assess and refer for IPV and related issues. The proposed study directly addresses this major clinical gap by testing whether the development and implementation of a digital, decision-support toolkit for IPV and co-occurring mental health conditions will increase standard screening for IPV, uptake of referrals, improve clinic workflows, and enhance clinical outcomes. The proposed study includes three separate aims, two in the R61 phase and a stepped-wedge randomized trial in the R33 phase. In Aim 1 (R61), informed by the Consolidated Framework for Implementation Research, we will conduct stakeholder interviews with providers, IPV advocacy specialists, and IPV survivors to inform the development of the digital toolkit to assess for IPV, PTSD, and associated conditions in OTPs. An iterative process of design for the digital toolkit will be employed. The decision-support algorithms will be developed to enhance automation of referrals following assessments. In Aim 2 (R61), we will beta test the digital toolkit with 12 provider-patient dyads in each participating OTP and further refine the digital toolkit. Go/No-Go criteria for the R33 phase will include quantitative and qualitative thresholds of acceptability, feasibility, appropriateness, and usability of the toolkit based on data provided from beta testers. If criteria are met, in Aim 3 (R33), we will apply the Reach, Effectiveness, Adoption, Implementation, and Maintenance (RE-AIM) framework to evaluate the digital toolkit in a randomized stepped-wedge, hybrid implementation-effectiveness type II design among N=780 individuals with OUD seeking care at one of the three OTPs in South Carolina. Drs. Jarnecke and Saraiya (MPIs) have assembled a team of renowned investigators with expertise in OUD, IPV, PTSD, clinical trials, digital interventions, and dissemination and implementation science. Findings could transform clinical practice by enhancing standard screening, assessment, and referral of IPV and associated conditions, and reduce IPV, PTSD, and OUD in the state of South Carolina, which has one of the highest rates of IPV in the U.S.

NIH Research Projects · FY 2025 · 2024-09

The burden of colorectal, cervical, and breast cancer cases and deaths is disproportionately greater among individuals living in impoverished areas. Early detection of these cancers through screenings and timely treatment among individuals diagnosed with precancers or cancers can reduce the excess cancer burden in these population subgroups. Actionable data to guide outreach efforts and implement programs and policies are urgently needed to reduce colorectal, cervical, and breast cancers among low-income communities. The National Cancer Institute (NCI) and the American Cancer Society (ACS) recently emphasized the promise of big data in improving knowledge and providing more accurate data for informing practice and policies. This project draws from the NCI’s and ACS’s vision of the strategic use of big data and integrates it within an interactive data visualization framework. We will utilize South Carolina’s Integrated Database System (covering health information of over 90% of low-income individuals) to determine colorectal, cervical, and breast cancer incidence and mortality rates among low-income individuals and develop a user-centered cancer data visualization tool through stakeholder engagement. Two end products (a web-based interactive tool and a toolkit) will be disseminated to community organizations and stakeholders working with low-income populations.

NIH Research Projects · FY 2024 · 2024-09

The use of exogenous nucleic acids has great potential for therapeutic applications in numerous diseases, including cancer. However, the delivery of nucleic acids is challenging, as they face numerous biological barriers, and thus require sophisticated delivery platform technologies. Despite the development of many promising non- viral carrier technologies, such as lipids, polymers, and peptides, achieving a clearer understanding of the mechanisms they employ to facilitate cell entry of nucleic acid-based drug cargos is, in-part, critical to advancing their clinical translation. As such, in our quest to further the development of a cell-penetrating peptide (CPP) carrier design for siRNA-based human oral and oropharyngeal cancer (OOC) therapy, we recently reported that a particular peptide variant, RD3AD, which exhibited enhanced siRNA uptake and gene silencing compared to the parent CPP, was found to direct siRNAs to specific cell-surface protrusions, identified as filopodia. Intriguingly, filopodia are highly dynamic, elongated, and thin cellular processes that have been reported to facilitate the highly efficient cellular entry of viruses, bacteria, exosomes, and other biological macromolecules/complexes. Moreover, because filopodia function as sensory antennae in probing the cellular environment, which is important for cell motility through complex 3D microenvironments, they contain cell-surface receptors, such as integrins, that can interact with and interpret extracellular cues. Interestingly, integrins have been reported to assemble along filopodia in similar localization patterns as RD3AD-siRNA complexes, thus implying that the RD3AD peptide carrier potentially targets integrins on filopodia as a means of cell entry. With filopodia having also been implicated in driving cancer cell migration and invasion, with their densities and lengths correlated with cancer progression and increased invasiveness, and increased levels of integrins also being associated with OOC and metastasis, the goal of the current proposal is to elucidate the mechanisms underlying filopodia-mediated intracellular uptake of the RD3AD-siRNA complex, as well as determine its ability to target and deliver siRNAs to migrating/invading cancer cells. To accomplish this goal, we will use biochemical and molecular biology approaches to: (1) uncover the mechanisms of filopodia-directed cell entry of the RD3AD- siRNA complex in oral cancer cells; and (2) demonstrate the ability of RD3AD to target and deliver complexed siRNAs to migrating/invading cancer cells and 3D tumor spheroids in vitro, as well as malignant tumors in vivo. The outcomes of the proposed high-risk, high-reward research are expected to uncover the mechanisms governing the intracellular delivery of the RD3AD-siRNA complex via its associations with filopodia and/or integrins, as well as its propensity to target migrating/invading cancer cells. The significance being that the knowledge gained will aid in future and more informed development of a CPP-based molecular targeted nucleic acid therapy for human OOC intervention.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT The objective of the proposed Mentored Patient-Oriented Research Career Development Award (K23) is to support Dr. Tanya Saraiya in acquiring the skills necessary to become an independent clinical researcher with a program of research focused on the modification and implementation of interventions for opioid use disorder (OUD) and comorbid mental health conditions, such as posttraumatic stress disorder (PTSD). Up to half of individuals with OUD have PTSD, but to date, there are no established evidence-based behavioral interventions that concurrently address OUD and PTSD symptoms. The proposed study directly addresses this major clinical gap by testing whether augmenting medications for OUD with an adapted, trauma-focused, integrated behavioral treatment for substance use disorders and co-occurring PTSD (i.e., Concurrent Treatment of PTSD and Substance Use Disorders Using Prolonged Exposure; COPE) will enhance clinical outcomes. In Aim 1, we will modify the existing 12-week, COPE intervention to individuals with OUD and PTSD using an iterative process informed by provider and patient feedback. In Aim 2, we will conduct an open-label trial (n = 5) to further refine and finalize the treatment protocol. In Aim 3, we will test COPE combined with medications for OUD (MOUD) versus MOUD-only in a randomized clinical trial among 76 individuals with OUD and PTSD to evaluate feasibility and preliminary efficacy in reducing opioid use and PTSD severity. During Aim 3, both groups will complete ecological momentary assessments (EMA) to assess for daily opioid use, craving, and PTSD symptoms. EMA will allow us to assess for the associations between daily PTSD symptoms and opioid craving and use, which may then inform treatment development. Dr. Saraiya has assembled a team of renowned mentors with expertise in OUD, PTSD, clinical trials, medications for OUD, and EMA. On-site mentors (Drs. Sudie Back and Kathleen Brady) have extensive knowledge in conducting clinical trials of behavioral and pharmacological interventions for co-occurring substance use disorders and PTSD. Off-site mentors (Dr. Katherine Mills at the University of Sydney and Dr. Kenzie Preston at NIDA) will provide guidance on how to modify the integrated intervention and leverage EMA methods to an OUD population. The mentorship and hands-on training afforded by the proposed K23 award will ensure that Dr. Saraiya achieves the following career goals: (1) a solid knowledge base in OUD and PTSD treatment; (2) proficiency in the design, evaluation, and implementation of clinical trials for OUD and PTSD; (3) experience with the implementation of EMA and skills in longitudinal analysis; (4) advanced training in the responsible conduct of research; and (5) stronger skills in manuscript and grant writing. The candidate will complete the proposed K23 activities at the Medical University of South Carolina, which has a strong track record of commitment to the advancement of early stage opioid use investigators. The proposed K23 activities will ultimately prepare Dr. Saraiya to lead a program of rigorously designed treatment research on OUD.

NIH Research Projects · FY 2024 · 2024-09

SUMMARY This proposal requests funds to acquire a high-performance computing (HPC) cluster with extensive data storage and Graphics Co-Processor (GCP) capabilities for the Medical University of South Carolina (MUSC) to support artificial intelligence (AI) based research integrating institutional laboratory, clinical, imaging and genomic data resources. MUSC is dramatically expanding its “raw materials'' for AI research: electronic health records (EHR) data from an expanding (now statewide) healthcare system; clinical and research-based imaging data stored in a common format in a new PACS (picture archiving and communications system); and deep genomic data (~100,000 over the next four years). These data create a unique opportunity along with a need for advanced HPC capabilities to expand existing research projects using these data resources. AI and specifically machine learning has led to groundbreaking biomedical studies, but requires HPC, robust graphics processing units (GPUs) and large accessible storage. Currently, a number of NIH-funded investigators at MUSC are performing research utilizing big data and machine learning, capturing massive amounts of data. Unfortunately, these highly valuable data sets are disjointed and siloed due to a lack of an adequate, unified data storage solution that can facilitate translational linking of these data sets. Growing unmet computational need is also a common theme among these NIH-funded investigators, particularly in areas of research involving high-volume, high-resolution image acquisition. Currently, inadequate access to high performance computing and the necessary AI infrastructure limit their ability to apply techniques such as deep learning to unlock a more complete value from these data sets. These data sets are often of a size and complexity such that it is impractical to transmit, store and compute upon them using cloud resources, requiring local HPC “instruments” to perform the computational tasks required. Building upon an existing MUSC institutional initiatives, proposal adds AI computational infrastructure to the socio-technical resources available at MUSC and to pilot level computational resources. The proposed HPC cluster will unify these efforts by providing common data storage with ample and accessible computing power for sophisticated AI techniques (e.g. deep learning) to exploit the potential synergies among data sets and facilitate translation efforts. The HPC cluster, while dedicated to research, will reside in the MUSC Data Center which serves the combined MUSC health and research enterprise.

NIH Research Projects · FY 2024 · 2024-09

The proposed scientific conference is organized by the International Society for Research in Human Milk and Lactation (ISRHML) Conference and entitled, “22nd ISRHML International Conference on Human Milk and Lactation Research: Advancing Science, Fostering Inclusivity, Shaping the Future” will take place on December 9-13, 2024, in Charleston, South Carolina. This conference will bring together researchers, clinicians, and healthcare professionals from diverse backgrounds and geographical regions around the globe, creating a dynamic platform for scientific exchange, education, and collaboration. The conference’s primary focus is to promote scientific excellence in the field of human milk and lactation research. Specifically, the Aims of the Conference are threefold: (1) Foster an inclusive exchange of scientific knowledge on the biology of human milk and lactation; (2) Advance cutting-edge research on the health effects of human milk and lactation; and (3) Translate basic science research into clinical practice and refine dissemination strategies. Distinguished from other conferences, this event is specifically designed to emphasize specialized research in human milk biology and composition and translating scientific discoveries from laboratory research to clinical practice. The conference will adhere to WHO code guidelines and offer CME/CE accreditation. Through this R13 application, we will offer trainee travel awards, virtual participation options, and accommodations for individuals with disabilities, ensuring that the conference is accessible to a broad audience. Beyond the conference, we are committed to evaluating the success of our inclusion, diversity, equity, and access (IDEA) efforts. Metrics include representation, attendee feedback, post-conference collaborations, longitudinal assessments, and comparisons to established benchmarks. These evaluations will inform our strategies for future conferences and contribute to creating a lasting impact on the scientific community. We anticipate sharing of breakthroughs, fostering new collaborations, and shared discoveries that will shape the trajectory of research in this vital field. Our Aims collectively underscore our commitment as a Society to advancing the scientific exploration of human milk and lactation, while emphasizing the global reach and inclusivity of our conference. By aligning our objectives with the goals of the NIH, through this planned conference, we aim to advance scientific knowledge, promote health equity, and ensure that research in human milk and lactation contributes significantly to the well-being of people worldwide.

NIH Research Projects · FY 2025 · 2024-09

Freezing of gait (FOG) is a common and debilitating manifestation of advanced Parkinson’s disease (PD) for which there are limited treatment options. Deep brain stimulation (DBS) of the subthalamic nucleus (STN) is effective in approximately half of patients with FOG (PwF), and its effects wane over time. We propose a multimodal neuroimaging study which uses diffusion MRI to understand the structural connections of the individual stimulation area, as well as the microstructural integrity of key nodes in the network. This study will also study the blood oxygen level dependent (BOLD) response to STN-DBS in PwF. The long-term goal of this research is to optimize FOG response to STN-DBS by identifying contributing modifiable factors. We propose to do so by: 1) studying differences in BOLD response to STN-DBS between responders and non-responders, 2) studying how the site of stimulation affects structural connectivity in responders compared to non-responders, and 3) studying differences in microstructural integrity of regions directly affected by STN-DBS (STN, GPi, PPN). Specifically, we aim to: 1) identify differences in BOLD activation based on FOG response to STN-DBS, 2) identify differences in structural connectivity to the stimulation site based on FOG response to STN-DBS and 3) identify differences in microstructural integrity of key network nodes. We will recruit PwF selected to undergo DBS surgery and perform structural imaging and behavioral assessments at baseline followed by combined DBS/fMRI studies and further behavioral assessments postoperatively and longitudinally. By achieving these aims we will have evaluated the contribution of lead placement, stimulation parameters, structural connectivity, and BOLD activation to FOG response which will be integrated to generate a mechanistic model of FOG response to STN-DBS. The proposed study is innovative in two major ways: 1) we propose a novel conceptual framework incorporating intrinsic and extrinsic factors that may affect FOG response to STN-DBS, and 2) we propose a novel approach which integrates structural connectivity with microstructural integrity along the circuit and to identify in-vivo functional network effects of STN-DBS activation in PwF. By developing a comprehensive integrated mechanistic model of STN-DBS response we can begin to develop optimization strategies to enhance engagement of the network. This approach will also further our understanding of the long-term therapeutic effects of STN-DBS by capturing longitudinal changes in functional network activation.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Myocarditis is a leading cause of heart failure and sudden cardiac death, but many cases of myocarditis follow a benign course. It is unknown what factors control this variable susceptibility to severe disease. Without this knowledge, the ability to develop targeted therapies to prevent disease progression is greatly limited. Viral infection is the most common known cause of myocarditis, and much research has focused on the pathways involved in the heart’s response to viral infection. Many such studies have identified type I interferon signaling as critical for viral pathogenicity and susceptibility to myocarditis. Quantitative trait locus analysis suggested the cardiomyocyte-specific protein cardiac troponin i3 kinase (TNNI3K) as a potential candidate. In an initial study, I infected homozygous Tnni3k knockout and control wild-type mice with coxsackievirus B3 (CVB3) for 10 days and demonstrated a direct role of Tnni3k in viral myocarditis. I additionally showed that Tnni3k’s kinase activity is necessary for mounting an effective response. Preliminary cell-based transfection of Tnni3k with an interferon signaling response element (ISRE)-linked reporter showed that presence of Tnni3k led to increased ISRE expression after treatment with interferon-α. Therefore, I hypothesize that TNNI3K decreases susceptibility to viral myocarditis in a kinase-dependent manner by enhancing the interferon response in cardiomyocytes. I will address this hypothesis by testing the role of Tnni3k in vivo (Aim 1) and in vitro (Aim 2). Aim 1 of this proposal expands on the initial study by infecting Tnni3k knockout and control wild-type mice with CVB3 and assessing measures of myocarditis after 4 days and after 35 days. Additional mouse models modifying the kinase activity of Tnni3k have been generated and will also be tested with viral infection. One of these models is of particular interest, as it recapitulates a common SNP found in millions of humans. In Aim 2, I will confirm the influence of TNNI3K in the interferon pathway by treating wild-type and TNNI3K knockout induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) with CVB3 and evaluating gene expression of proteins in the interferon pathway. I will then probe for differentially phosphorylated proteins in wild-type and TNNI3K knockout iPSC-CMs following CVB3 infection. Differentially phosphorylated proteins will be silenced in iPSC-CMs to define their role in the interferon pathway specifically in cardiomyocytes. Results from this study will document a mechanism by which TNNI3K controls susceptibility to viral myocarditis. This model will enable future studies aimed at developing novel therapies targeting myocarditis progression at the molecular level. This fellowship award will support my development as an exceptional cardiologist-scientist.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Adolescent alcohol misuse is a significant risk factor for alcohol use disorder in adulthood. Particularly concerning are the long-term effects on brain maturation and associated cognitive function, including the ability to appropriately process and respond to pain. A growing body of research indicates that adolescent alcohol misuse enhances pain sensitivity and anxiety in adulthood, and increased pain sensitivity and heightened anxiety may increase the risk for alcohol misuse later in life. Recently, a role for a circuit involving the basolateral amygdala (BLA), prelimbic (PrL) cortex, and ventrolateral periaqueductal gray (vlPAG) in pain processing has been described. Within this circuit, activation of the BLA by nociceptive stimuli results in increased feedforward inhibition of PrL neurons projecting to the vlPAG (PrLPAG). This feedforward inhibition is mediated by parvalbumin interneurons (PVINs) in the PrL. Within this circuit, PVINs modulate pain sensitivity and pain-related affect by regulating the activity of PrLPAG neurons. Adolescence is a critical period for development. Environmental insults occurring during adolescence, such as those caused by repeated episodes of binge-like alcohol consumption, disrupt normal development of PrL circuitry. These changes include reduced intrinsic excitability of and evoked AMPA and NMDA mediated currents onto PVINs. In addition, preliminary data we have collected indicates that following adolescent intermittent alcohol exposure the excitatory/inhibitory (E/I) balance evoked by optogenetic stimulation of BLA terminals in the PrL cortex is reduced at PrL PVINs and increased at PrLPAG neurons. Further, following alcohol exposure during adolescence, the density of perineuronal nets (PNNs) which enwrap PVINs and restrict plasticity is increased in the PrL. The overarching hypothesis of this proposal is that persistent changes in synaptic function following AIE are associated with altered activation patterns of PrL PVINs and PrLPAG neurons that are correlated with altered pain sensitivity. We further hypothesize that digestion of PNNs using chondroitinase ABC will reverse changes in the E/I balance at PrL PVINs and PrLPAG neurons. The proposed studies will use a combination of cell-type and projection specific chemogenetic manipulations to determine the effect of changes in the activity of PrL PVINs and PrLPAG neurons on nociception and pain-related affect following adolescent alcohol exposure. Changes in the activation patterns of PrL PVINs and PrLPAG neurons in response to nociceptive stimuli will also be assessed using fiber photometry. Finally, to determine whether PNNs stabilize altered synaptic function following adolescent alcohol exposure, PNNs will be digested and the E/I balance at BLA inputs onto PVINs and PrLPAG neurons will be measured using acute slice electrophysiology. These experiments will provide valuable insights into how changes in PrL PVIN and PrLPAG synaptic function resulting from adolescent alcohol exposure affect pain threshold in adulthood.

NIH Research Projects · FY 2026 · 2024-09

The initiators and/or drivers of Age-related Macular Degeneration (AMD) remain elusive. Current knowledge suggests that interventions in late disease stages may not be as effective as earlier in the disease due to a “domino effect”, wherein there are many inflammatory factors acting later in the disease. We posit that there is value in doing proof of mechanism studies to identify biological effects at the early stage of disease. The major genetic risk factors include CFH on chromosome1 and ARMS2/HTRA1 on chromosome 10. Despite genetic heterogeneity, most subjects with AMD show common early pathology that include drusen formation and complement activation. Based on these common features, the core structures affected in AMD that require further investigation are the RPE, Bruch’s membrane (BrM) and the choroid. The recent discovery of induced pluripotent stem cell (iPSC) tissue models to study disease mechanisms and the ability to assemble 3D tissues permit the study of cell-cell interactions. We have designed a 3D tissue chip which includes an RPE monolayer sitting on a choroid containing endothelial cells, fibroblasts and pericytes which eventually form a BrM to complete the structure. Since the endothelial cells undergo anastomosis the final choroidal vasculature can be perfused to more closely mimic the in vivo environment. Our current goal is to utilize this model to characterize the responses of the RPE-choroid to various environmental stressors under the influence of the CFH and ARMS2/HTRA1 risk genotypes. Please note, not only can we assemble the 3D oBRB from different donors, but we can subsequently analyze four different parts; the supernatant in the top compartment (i.e., sub-retinal space), the flowthrough in the bottom compartment (i.,e, blood supply of the choroid), as well as the two tissue compartments, RPE/BrM and CC. Aim 1 will characterize the perfused 3D RPE/choroid chip and provide baseline data for our readouts (gene expression patterns based on scRNA-Seq; extracellular and intracellular complement activation; BrM formation over time; and extracellular vesicle analysis). In Aim 2, we ask how these parameters are affected genetic risk factors, assembling the RPE/choroid complex from iPSC cells using a combination of RPE-risk with choroid-non-risk or vice versa. And finally, in Aim 3, we characterize the response of the perfused 3D RPE/choroid chip to AMD relevant stressors, smoke, fatty acid exposure and alteration in flow rate. Overall, our data will provide unique knowledge about the influence of genetic variation on complement secretion, expression and synthesis of intracellular and extracellular complement from pre- drusen to post-drusen development, and the proposal’s outcomes will fill a critical gap in our understanding of genetic variants on RPE complement activity and identify new potential therapeutic targets directed at the RPE in the early stages of AMD.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Depression is the most common neuropsychiatric manifestation following stroke and current treatments are largely ineffective. Depression has both direct and indirect effects on response to rehabilitation treatment, thus subjects with post-stroke depression (PSD) are routinely excluded from rehabilitation trials and treatment options for these individuals are extremely limited. We propose to determine the efficacy of combining two known anti-depressant treatments shown to be effective in non-stroke depression, aerobic exercise (AEx) and repetitive transcranial magnetic stimulation (rTMS), on post-stroke depressive symptoms. Further, we will determine 1) the potential for AEx to enhance the effects of rTMS to treat depression; and 2) whether successful reduction in depressive symptoms is associated with improved recovery. This project is based on the scientific premise that depression negatively affects the potential to adapt in response to treatment such that rehabilitation may not produce the same changes that it does in non- depressed individuals. We hypothesize that effective treatment for PSD will result in a virtuous cycle whereby reducing depression enhances response to rehabilitation, thereby facilitating functional gains. That is, effectively treating depression will enable individuals to better recover from stroke. Furthermore, in addition to its beneficial effects on depression, AEx is known to improve post-stroke motor recovery, thus providing an attractive option for treating depression as well as an established vehicle to study the effects of PSD on response to rehabilitation. The experiments proposed as part of this project are designed to address critical questions related to: 1) the combined effects of AEx and rTMS (versus AEx or rTMS alone) on depressive symptoms; 2) the potential for PSD to limit response to rehabilitation; and 3) the relationship between baseline subject attributes and response to treatment.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Alcohol use disorder (AUD) is a devastating condition that affects 29.5 million people aged 12 and older. AUD is classically characterized by a loss of behavioral control, particularly in the face of negative consequences, resulting in a loss of inhibitory control, or disinhibition, that may normally suppress alcohol seeking. Considering this, it is critical to investigate mechanisms associated with disinhibition. One major barrier to investigating AUD progression is understanding specific cell populations that may normally serve to suppress alcohol seeking but become disinhibited following dependence. While our knowledge of neuronal ensembles in alcohol dependence remains limited, studies on the paraventricular thalamus (PVT) projection to the nucleus accumbens (NAc) in regulating sucrose seeking have revealed distinct neuronal ensembles that emerge across the course of learning. These studies indicate that PVTàNAc encodes ensembles of inhibited neurons that function as a “brake” on reward-seeking. Extending these findings to examine how alcohol influences thalamo-striatal neuronal ensembles is imperative for targeting inhibitory ensembles that may attenuate alcohol-seeking behaviors. My preliminary data suggest that broad optogenetic activation of PVTàNAc neurons reduces alcohol-seeking, evidenced by decreased active lever presses and alcohol deliveries during an operant head-fixed task in nondependent mice. Conversely, optogenetic inhibition of PVTàNAc neurons releases this “brake” and promotes active lever pressing, even in the face of behavioral suppressors. Importantly, after inducing alcohol dependence via chronic intermittent ethanol (CIE) exposure, presentation of behavioral suppressors, including direct activation of PVTàNAc neurons, no longer reduces alcohol seeking, indicative of a functional uncoupling of this circuit leading to behavioral disinhibition. These current data suggest that PVTàNAc neurons may serve as an exciting new target for studying how chronic alcohol induces disinhibition at the level of neuronal ensemble activity. To date, no work has explored single-cell manipulation or two-photon calcium imaging of PVTàNAc neuronal ensembles in regulating alcohol seeking before and after alcohol dependence. The focus of this proposal therefore, is to identify and manipulate specific PVTàNAc neuronal ensembles that may be altered after dependence. My overarching hypothesis is that inhibitory neuronal ensemble activity in PVTàNAc will best decode alcohol-seeking behaviors and become persistently inactivated following ethanol dependence reducing feedforward inhibition onto PV-INs. Aim 1 will measure calcium dynamics of PVTàNAc neurons in vivo to identify unique neuronal ensembles that emerge during alcohol seeking. I will then manipulate specific PVTàNAc neurons that may be altered before and after alcohol dependence. Aim 2 will assess how inputs from PVT onto parvalbumin neurons in the nucleus accumbens may be altered following dependence ex vivo. Collectively, this research will provide valuable and novel information on the role of thalamo-striatal ensembles to enhance our understanding of underlying mechanisms of disinhibition in AUD.

NIH Research Projects · FY 2025 · 2024-09

Project Abstract: The human gastrointestinal tract houses a diverse community of commensal gut bacteria that actively participate in maintaining host health. Among the intricate and dynamic relationships within this ecosystem, the interaction of bacteria with mucus, a crucial protective barrier lining the gut, emerges as a vital and multifaceted process. Mucus, primarily composed of mucin glycoproteins, acts as a nutrient source and signaling platform, playing pivotal roles in microbe- host interactions. While it is well known that gut bacteria can degrade mucus and cross-feed non- mucus degrading microbes, the intricacies of this interaction remain largely unknown. Additionally, the ability of intestinal mucus to regulate bacterial communication is not clear and the reciprocal interaction of how bacteria regulate mucus production is not well understood. In many ways, we are only just beginning to understand how mucus influences bacterial ecology and bacterial-host interactions. This proposal addresses these challenges using the expertise of the Engevik lab, resources and reagents we have generated to study bacterial-host interactions, and the expertise of our collaborators. The overall goal of the research program is to unravel the intricate mechanisms by which commensal gut microbiota respond to mucus and stimulate mucus production by the host. The Engevik Lab opened at MUSC during the beginning of the pandemic. The Engevik lab pursues key questions: What bacterial compounds are a direct result of mucus degradation? How do bacteria grow in communities in the setting of intestinal mucus? What aspect of bacterial signaling does mucus regulate? What metabolites do bacteria produce to increase mucus production? Our multidisciplinary approach encompasses cutting-edge techniques in microbiology, metabolomics, transcriptomics, and host-microbe interaction studies to comprehensively address these questions and delve deeper into the bacterial ecology in the gut. Insights gleaned from this research have the potential to pave the way for innovative approaches to enhance gut health and mitigate diseases linked to disruptions in the gut microbiome. Ultimately, this investigation aligns with the NIH's mission to foster groundbreaking discoveries that promote human health and well-being.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Age-related speech recognition difficulty and its association with poorer memory, increased fatigue, social isolation, and cognitive decline are major public health concerns with the current aging population. Older adults consistently show increased listening difficulties compared to younger adults, including poorer understanding and memory for speech in noise. The long-term objective of this research is to investigate age-related brain differences that affect (1) how quickly and (2) how much information is used during the perceptual recognition of speech in noise. We will use a perceptual decision-making framework to characterize differences in the speed of evidence accumulation and height of decision criteria, which can affect the accuracy and latency of speech recognition. Older adults appear to accumulate evidence more slowly and use more cautious decision criteria than younger adults. Brain imaging studies have linked evidence accumulation and decision criteria- setting to distinct brain systems, which also decline with older age. However, the contribution of perceptual decision-making to age-related listening difficulties is not yet known. The interrelated Aims 1-2 for the current research will test the overarching hypothesis that age-related declines in perceptual decision-making processes limit understanding and memory for speech in noise, commonly observed for older adults. The Aim 1 neuroimaging experiments manipulate how quickly evidence accumulates, to characterize the degree to which older adults accumulate evidence more slowly than younger adults based on uncertain and noisy sensory information. We predict that slower evidence accumulation measures are associated with poorer understanding and delayed recognition memory for older adults compared to younger adults. Structural diffusion imaging data will be collected for white matter tracts to determine whether declines in the connections between brain regions are associated with slower evidence accumulation. The Aim 2 neuroimaging experiments manipulate decision criteria, to characterize age-related differences in how much evidence is collected in challenging listening conditions. Older adults often respond more slowly and cautiously in noisy conditions compared to younger adults, regardless of task difficulty. Older adults also demonstrate poorer memory for targets and better recognition of distractors than younger adults. We predict that decision criteria variation will be associated with poorer understanding and memory for speech in noise. Structural gray matter measures will be collected to examine neural declines accounting for age-related differences in decision criteria, given that frontal brain regions linked to criteria show lower volume for many older adults. Perceptual decision-making can explain differences in speech recognition performance outside of the neuroimaging environment, so this research is a critical first step to developing measures to characterize an individual's listening difficulties and guiding effective rehabilitation strategies.

NIH Research Projects · FY 2025 · 2024-08

The COVID-19 pandemic has greatly exacerbated rising patterns of excessive alcohol consumption, particularly in women. Binge alcohol drinking is the most common pattern of excessive alcohol drinking and is associated with increased risk of developing mood disorders and Alcohol Use Disorder (AUD). Social support strongly buffers against alcohol craving and relapse, yet many individuals display reduced valuation of social rewards and/or deficits in the processing of social stimuli following alcohol exposure and subsequent abstinence. This may compound interpersonal problems in people with AUD and limit their capacity to seek out or receive social support. Therefore, understanding the neurobiological mechanisms mediating alcohol’s effects on social behavior is required to inform future treatments aimed at enhancing social functioning and reducing relapse in patients with AUD. Drinking in the Dark (DiD) is a robust paradigm for investigating the circuit and molecular mechanisms of binge-like alcohol consumption on physiology and behavior in rodents. In addition, the 3-chamber sociability model permits the interrogation of both social reward and social recognition behaviors following alcohol consumption. Serotonin (5-Hydroxytryptamine, 5-HT) receptor signaling in the lateral habenula (LHb) has been implicated in the development of negative emotional states associated with abstinence from alcohol, but the LHb remains highly understudied in the context of AUD. I recently found that DiD reduced social recognition selectively in female mice during abstinence, and that genetic deletion of the LHb Gq-protein coupled serotonin receptor 5HT2c partially prevented this effect. My preliminary data suggests that binge alcohol consumption enhances the intrinsic excitability of LHb 5HT2c-containing neurons (LHb5HT2c) in the medial sub- region, that 5-HT is released onto LHb5HT2c during social interaction, and that acute engagement of Gi signaling in LHb5HT2c can normalize social deficits induced by alcohol. Moreover, DiD appears to modulate the expression of multiple 5-HT receptor subtypes co-expressed in LHb5HT2c. Together, these data suggest that excessive activation of LHb5HT2c via dysregulation of 5-HT receptor signaling may underlie social deficits induced by alcohol. Using these preliminary findings as a foundation for the current proposal, I will 1) characterize how DiD alters the translational and physiological landscape of LHb5HT2c projecting to the DRN (5HT2cLHb-DRN) and 2) investigate the functional impact of genetic manipulation of 5-HT receptor sub-types in 5HT2cLHb-DRN on neuronal physiology and DiD-induced dysregulation of social behavior. Together, these experiments will determine the effects of binge alcohol drinking on molecular and physiological processes in 5HT2cLHb-DRN and identify novel mechanisms by which these neurons promote social dysfunction during abstinence. Furthermore, this award will provide me with valuable technical and professional training that will facilitate my transition to independence.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Fibroproliferative illnesses leading to organ fibrosis and failure are responsible for approximately 45% of deaths in developed countries. Whether idiopathic, triggered by environmental factors, infections, or genetics, organ fibrosis results in significant morbidity and mortality. Organ fibrosis is responsible for health care costs exceeding $10 billion/year. It is estimated that the number of deaths due to fibrosis is double the number of deaths due to cancer, and that organ fibrosis results in significant physical, emotional, and financial burdens. Specifically, lung fibrosis can be idiopathic, associated with connective tissue diseases, or triggered by environmental and occupational exposures such as radiotherapy. There are currently no effective therapies to treat existing lung fibrosis as recently approved drugs merely reduce disease progression and result in significant side effects. Thus, the only curative option for patients is organ transplantation, which is impossible at the scale needed. We have identified peptides derived from collagen XVIII which exert anti-fibrotic effects in murine and human pre- clinical models of lung fibrosis. The beneficial effects include reducing fibrosis in lung tissues of patients with pulmonary fibrosis who underwent lung transplantation and thus have end-stage severe fibrosis, an effect not seen with other drugs that are approved or being evaluated for these illnesses. We propose to simultaneous test the identified peptides in our pre-clinical models of fibrosis and identify a lead candidate. We also propose to further characterize the efficacy of the lead candidate, conduct dose escalation studies, and optimize the dosing regimen. We have assembled a unique team with the expertise to develop the lead peptide. Our team includes an accelerator partner. Successful completion of this project will support our long-term goal of translating our findings to the clinic and provide patients with pulmonary fibrosis with an effective therapy. Our approach is likely to have broad impact and relevance for fibrosis in different organs.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Research on the microbiome-brain axis indicates strong bidirectional communication systems between the microbiome (including the oral microbiota) and the brain. In adults, the microbiome-brain axis is implicated in the development and maintenance of alcohol use disorder (AUD) and has been examined as a potential AUD treatment target; however, similar research is lacking during adolescence. In our recent review of the adolescent oral and gut microbiota, we found that the adolescent microbiome may be vulnerable to alcohol use. It is important to investigate if the microbiome-brain axis findings from the adult literature can be replicated in adolescents who use alcohol, or if the developmental aspect of adolescence incurs differential effects. The genus Lactobacillus is of particular interest in the adult AUD field and provides an ideal initial target for extending microbiome-brain research into adolescence. Lactobacillus shows a lower relative abundance in relation to adult alcohol use; has been proposed as a treatment option for adult AUD; and administration in a preclinical model resulted in brain metabolite alterations (glutamate, GABA, and N-acetylaspartate or NAA) measured with proton magnetic resonance spectroscopy (MRS). Combined, these findings indicate that alcohol related alterations to Lactobacillus may led to brain metabolite level alterations through the microbiome-brain axis. Investigating the microbiome-brain axis during adolescence will provide insights into a critical piece of the AUD trajectory, as well as potential prevention (e.g., vulnerability markers) and intervention (e.g., pre/probiotics) targets. The overarching hypothesis is that the adolescent microbiome-brain axis is impacted by alcohol use. This application proposes an initial assessment of this hypothesis through novel research that will collect (1) salivary samples to assess the oral microbiome, with Lactobacillus as the primary genus of interest, and (2) Glu, GABA, and NAA brain metabolite levels within the dorsal anterior cingulate cortex via MRS. The oral microbiome was selected for this age range due to increased feasibility and conserved scientific integrity (e.g., resembles the upper gastrointestinal tract and sensitive to alcohol). We will recruit 126 adolescents (ages 14-18) with no/low alcohol use (n=42), recent moderate alcohol use (n=42), or recent heavy alcohol use (n=42). The proposed research plan in this NIAAA K01 application will serve as essential hands-on training to promote Dr. Kirkland’s career development in adolescent alcohol use through five distinct career objectives: (1) become proficient in oral microbiome data collection, analysis, and interpretation; (2) develop foundational knowledge of the adolescent microbiome, specifically its development, the potential effects of alcohol, and its connection to the central nervous system through the microbiome-brain axis; (3) gain experience in lab management skills to foster independence; (4) continue training in responsible conduct of research; and (5) enhance scientific writing, grant writing skills, and science communication skills. The mentorship and research skills gained through this application will set Dr. Kirkland up to be a successful and prolific NIH-supported independent researcher.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Respiratory data from five existing human subject datasets will be merged in order to comprehensively examine the dynamic and coordinated movements of the chest wall system (thorax and abdomen) in persons with Parkinson’s disease (PD). The precisely coordinated movements of the chest wall are critical for optimizing speech production and minimizing the work of breathing. One of the most salient speech and voice symptoms associated with PD is a reduction in speaking volume, a condition known as hypophonia. Hypophonia is estimated to occur in 80-89% of persons with PD and leads to social isolation and depression due to a significant adverse impact on spoken communication. Hypophonia has been attributed to several physiological factors including reduced ribcage compliance, decreased abdominal support, and reduced vocal fold closure. Asynchronous movement of the chest wall system, however, may be another contributing factor, which is presently not accounted for in our voice intervention protocols. It is widely reported that persons with PD exhibit a myriad of motor coordination issues, stemming from basal ganglia and cerebellar dysfunction, including impaired fine motor dexterity, postural instability, and abnormal gait characteristics. An empirical study of the dynamic and coordinated movements of the chest wall system during breathing, however, is lacking in persons with PD. To date, studies have described pulmonary restriction in persons with PD using static measures of respiratory function (e.g. respiratory muscle strength and vital capacity). There is a critical need to study the dynamic movement of the respiratory system in order to develop effective and efficacious approaches to voice intervention. The present application seeks to address this paucity of data through a novel exploration of thoracoabdominal asynchrony (TAA) and paradoxical motion in a large sample of persons with PD (n=110). Small-scale studies on TAA and paradoxical motion suggest a high prevalence in persons with PD during rest breathing. The present application will examine the dynamic movement of the thoraco- abdominal system across the inspiratory and expiratory cycles of rest and speech breathing (Specific Aim 1) using standardized measures from the literature. Targeting TAA during speech breathing is innovative as this has not been studied in any population yet. Further, this study will examine how current voice intervention protocols, which directly or indirectly target respiration, impact the coordination of the thoraco-abdominal system during speech production (Specific Aim 2). Our application addresses NHBLI Strategic Objective 5: “To develop and optimize novel diagnostic and therapeutic strategies to prevent, treat, and cure National Heart, Lung, and Blood (NHLB) diseases.” The knowledge gained from this study has the power to transform our approach to voice and respiratory interventions for persons with PD and lead to efficacious and potentially new treatment protocols.

NIH Research Projects · FY 2026 · 2024-08

ABSTRACT Hearing impairment in children with autism spectrum disorders (ASDs) may contribute to core ASD symptoms by interfering with language development and communication. The number of individuals with ASD or other neurodevelopmental disorders is growing rapidly, thus studies are urgently needed to address how and to what extent auditory system deficits are associated with ASD-like behaviors. Mounting evidence supports the hypothesis that peripheral auditory system deficits reflective in auditory nerve dysfunction may contribute to the pathophysiological changes observed at the cortical level (e.g., hyperacusis or other auditory processing deficiencies). However, the lack of mechanistic studies using translational animal models of ASD with well- characterized auditory functional impairments is a critical barrier to uncovering potential relationships and causal links between structural and functional auditory deficits and ASD-related behaviors. Mutations or deletions in the Myocyte-specific Enhancer Factor 2C (MEF2C) gene have been linked to ASD and other neurodevelopmental disorders in humans. Our recent studies using a mouse model of human MEF2C haploinsufficiency syndrome (MCHS) revealed that Mef2c deficiency leads to altered neural activity in the cortex, increased microglial activation, and deficits in communication, social interaction, and other MCHS symptoms that are characteristic of ASD. Using this mouse model, our studies also show (1) that Mef2c may play a role in the development and function of spiral ganglion neurons of the auditory nerve and regulation of cochlear macrophage activity, (2) an important association between Mef2c deficiency and auditory nerve dysfunction, and (3) that Mef2c deficiency leads to cortical neural dysfunction in response to auditory stimuli. Together these exciting data support the hypothesis that ASD risk-gene MEF2C deficiency in spiral ganglion neurons and macrophages can lead to auditory nerve dysfunction and increased macrophage activation, which in turn contributes to auditory cortical dysfunction and core symptoms of ASD. This multiple PI proposal will test this novel hypothesis using a multidisciplinary approach. The proposed experiments will characterize unique temporal and spatial expression patterns of Mef2c in both spiral ganglion neurons and cochlear macrophages that are associated with auditory nerve development and maintenance (Aim 1), determine the causal links between Mef2c deficiency-induced auditory nerve dysfunction and auditory cortical dysfunction, and core symptoms of ASD (Aim 2), and test the extent to which inhibition of abnormal macrophage activation ameliorates auditory nerve dysfunction, auditory cortical dysfunction, and core ASD symptoms (Aim 3). These investigations will promote a greater understanding of the important role of immune cells and auditory system deficits in ASD and possible other neurodevelopmental disorders and may reveal a neuro-immune-based therapeutic strategy beneficial for this increasingly common disorder.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY Whole-genome duplication (WGD) is increasingly appreciated as a defining moment for enabling chromosome instability in solid tumors. This proposal investigates chemotherapy-induced WGD that promotes the formation of polyploid cancer cells that appear senescent but in the absence of functional p53 are able to escape the senescent state and generate heterogenous daughter cells. The focus on high-grade serous ovarian carcinoma (HGSOC) derives from the urgent clinical need to improve therapy outcomes and the fact that p53 mutations are the most prevalent genetic abnormality (≥90%) associated with aneuploidy in this malignancy. Polyploid cancer cells exhibit a high level of plasticity and are thought to be a driving force in repopulating a tumor with highly aggressive, therapy-resistant, metastasis-prone progeny. Currently, there is a gap in knowledge that connects clinical descriptions of polyploid cells in tumor specimen with laboratory observations on the formation of polyploid cancer cells. We hypothesize to close this gap through a CRISPR-Cas9 screen to identify genes that are functionally necessary for formation of polyploid cancer cells and their daughter cells. We further hypothesize that polyploid cancer cells in clinical specimen can be identified by their large polynuclear morphology and one or more putative markers (cytoplasmic p21, acid ceramidase, VEGF-R2) resulting from an unbiased RNAseq analysis. Our specific aims are to (1) Define essential genes for the polyploid lifecycle critical transition stages and (2) to determine expression of putative markers of polyploid cancer cells in archived HGSOC samples. The proposed studies will serve as the foundation for future larger mechanistic, drug development, or prognostic studies with the potential to improve outcomes in ovarian cancer patients. An immediate impact will include prioritizing patients for the angiogenesis inhibitor Bevacizumab, a common HGSOC option, which may work better in VEGF-R2 expressing PGCC tumors. Of particular interest will also be therapeutic targeting of the polyploid population through re-purposing of existing drugs that would be combined with neo-adjuvant chemotherapy or used as a maintenance strategy following surgical debulking. Results obtained in HGSOC likely have broader relevance, since polyploid cells are observed in many other solid malignancies.

NIH Research Projects · FY 2026 · 2024-08

Abstract Cellular therapies for the treatment of autoimmune diseases offers a novel treatment option with possible increased efficacy and decreased toxicity. We are performing the first multi-center double blind placebo controlled dose escalation trial of mesenchymal stromal cells (MSCs) for standard of care refractory lupus. This Phase II trial is based on promising data from open label trials in China and our own Phase I trial. Response rates very from 65-75% in reports from China while we saw an SRI-4 response in 5/6 patients we treated with umbilical cord derived MSCs. There are nine US centers participating in the MiSLE trial. Due to impact of the COVID-19 pandemic, we were unable to reach our goal of 81 patients during a previously funded NIAID U01 grant that expired in March, 2023. Sixty two of the 81 patients are either randomized or treated as of now. We were meeting our benchmarks of 1.98 patients per month until a COVID induced shutdown the trial for months. We are recruiting patients at this rate again. In the trial to this point, we have had no safety signals, no issues with infrastructure and patients meeting the primary study endpoint of the SRI-4. This is a critical trial to finish to definitely show if there is a treatment effect of MSCs in lupus and to delineate the mechanisms by which MSCs impact disease. In the Phase I trial we found marked impacts on extra follicular derived B cells. We are seeing similar trends in preliminary analysis of the Phase II trial, though we are blinded as to treatment group. We have introduced novel methods of multi-parameter statistical analysis to enhance detecting a treatment effect. In mechanistic studies we will complete the analyses of B and T cell subsets, receptor diversity and epigenetics over the course of the trial. Based on the development of new technology, we will perform CyTOF on mononuclear cell populations, Cite Seq single cell analysis and plasma microbiome assays. These assays will be performed at weeks 0, 4, 8 and 24. We have sufficient stored samples to perform the proposed assays. This highly impactful trial will provide important insight into changes over time of key immune mediators of lupus, perhaps allowing pre-treatment identification of patients most likely to respond to MSC treatment.