Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2022-09

This K01 career development proposal was developed to support Dr. Shakia Hardy, a tenure track Assistant Professor in the Department of Epidemiology at the University of North Carolina at Chapel Hill (UNC), in her path towards becoming an independent researcher. The Candidate earned her PhD in Epidemiology in 2017 from UNC and has a strong record of published research in the fields of cardiovascular and social epidemiology. Her goal is to integrate her epidemiology skills with implementation science to develop an independent research program at the intersection of implementation and population science. Dr. Hardy’s research aims to reduce health disparities in cardiovascular disease through the modification of lifestyle behaviors that contribute to increased blood pressure in early life. Training: To accomplish her overall goal, Dr. Hardy has proposed an intensive mentored research plan that includes didactic education, and experiential learning in 1) intervention development, 2) implementation science for intervention delivery and evaluation, 3) design and conduct of randomized controlled trials, and 4) leadership skills for academic scientists. Mentorship: An interdisciplinary team of renowned scholars will mentor Dr. Hardy and provide ongoing guidance as she transitions to independence. Her mentoring team includes a Primary Mentor, Dr. Andrea Cherrington, co-Mentors, Dr. Paul Muntner, Dr. Nathalie Moise and Dr. Daniel Feig, and two content advisors. Each of these mentors are fully committed to this project and to Dr. Hardy’s success. Research: The prevalence of high BP, defined as systolic BP (SBP) ≥120 mm Hg or diastolic BP (DBP) ≥80 mm Hg, is twotimes higher (20% versus 10%) among Black versus white adolescents 13-17 years of age in the US. The availability of lifestyle behavior counseling in a clinical setting to lower BP is limited in rural communities due to long travel distances to access healthcare, limited funds for copayments, and ineffective provider-patient communication. The proposed K01 addresses this gap by developing and testing the feasibility and acceptability of a peer support intervention to lower BP among rural Black adolescents with high BP. First, Dr. Hardy will identify aspects of the rural environment that present barriers to lifestyle behavior change. Next, using the behavior change wheel for intervention development, she will develop and pilot test a peer support intervention devised to address these barriers. The successful completion of this project will lead to an R01grant submission to test the effectiveness of this peer support intervention in a multi-site clusterrandomized trial and allow Dr. Hardy to become an independent investigator contributing to reducing health disparities among adolescents, where lifestyle behavior change could preempt the development of racial disparities in BP and cardiovascular morbidity and mortality.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/ Abstract: Lymphatic diseases are numerous and affects upwards of 200 million people worldwide. These chronic disorders have limited clinical management and are broadly characterized by aberrant lymphatic vessel development and/or dysfunction which results in painful accumulation of interstitial fluid. Shockingly, no FDA-approved pharmacological treatments targeting lymphangiogenesis, the process of lymphatic vessel formation, are available. Thus, there is an urgent need to characterize key therapeutically tractable proteins and signaling pathways that regulate lymphangiogenesis. The Caron laboratory studies one such molecule, the potent pro-lymphangiogenic peptide, adrenomedullin (AM). AM-induced lymphangiogenesis requires formation of well-controlled AM-chemotactic gradients to provide directionality to growing and migrating lymphatic vessel tips. The atypical chemokine receptor 3 (ACKR3) is critical for the establishment of these gradients through the internalization and degradation of AM. Recently, the Caron laboratory identified a novel interaction between ACKR3 and receptor-activity-modifying protein 3 (RAMP3). They showed in vitro that RAMP3 is required for the recycling of ACKR3 to the plasma membrane after AM-stimulated internalization and that loss of ACKR3 or RAMP3 in vivo results in impaired vascular development. However, the mechanism by which RAMP3 regulates ACKR3 activity and signaling in lymphatic endothelial cells (LECs) and the process of lymphangiogenesis remains unknown. RAMP3 is unique among the RAMPs in that it contains a C-terminal PDZ motif that mediates its function by promoting protein-protein interactions with PDZ domain-containing proteins. Therefore, the overarching hypothesis of this training proposal is that RAMP3 and its PDZ motif enhances ACKR3 activity and signaling within LECs to regulate lymphangiogenesis. Completion of this proposal will define the role of RAMP3 in the regulation of ACKR3 signaling and lymphangiogenesis, thereby advancing the current knowledge of the lymphatic biology field. In addition, this proposal will provide invaluable experience and training in the performance of ethical and rigorous research and effective scientific communication.

NIH Research Projects · FY 2025 · 2022-09

Abstract. The integral roles of histone post-translational modifications (PTMs), including lysine methylation and acylation, in chromatin remodeling is well established, but new roles of histone PTMs in disease are continually discovered. This rapidly evolving field necessitates the development of new molecular approaches to probe the biological impacts of these PTMs. Herein we focus on the reader proteins for lysine methylation and acylation as reader proteins are responsible for sensing the PTM and triggering the biological outcome, and both r have biomedical relevance. Our research will encompass development of new chemical biology tools to investigate binding mechanisms, improve detection of PTMs, and to generate designer nucleosomes to study the role of dual PTMs in biological pathways. This research is expected to advance the ability to develop selective inhibitors and molecular probes for these two classes of proteins. To develop new methods for creating new sensing methods and designer nucleosomes, we will exploit the inherent selectivity of reader proteins to develop engineered reader proteins (eReaders and eWriters) for sensing, labeling, and introduction of dual PTMs in a site selective manner. This combined effort will advance the field by providing new insight and new tools to study the biological pathways that depend on these PTMs and their role in disease.

NIH Research Projects · FY 2024 · 2022-09

ABSTRACT The failure of traditional mental health services to significantly reduce the disproportionate involvement of people with serious mental illnesses (SMI) in the criminal justice system highlights a basic but overlooked fact: mental illness is not a primary driver of criminal justice involvement. Research has consistently shown that people with mental illness face the same risk factors for justice involvement (i.e., “criminogenic” risk factors) as those without mental illnesses. Research has also found that justice-involved people with SMI have high levels of criminogenic risk factors and that these risk factors mediate their risk of recidivism. Yet, most interventions for justice-involved people with SMI do not target criminogenic risk factors as a goal of treatment. This gap between research and service provision represents untapped potential to reduce rates of justice involvement among people with SMI, by expanding their continuum of services to include interventions that directly target criminogenic risk factors. Our recent NIMH-funded randomized controlled trial (1R34MH111855-01; PI: Wilson) demonstrated that when adapted for use with people with SMI, criminogenic-focused interventions can successfully engage key treatment targets and outcomes associated with criminogenic risk factors among prisoners with SMI. Given that up to 50% of people with SMI receiving treatment in the community-based mental health system have had some criminal justice system involvement, developing criminogenic-focused interventions for delivery in community mental health settings has great potential to optimize their potential impact, both in terms of the numbers of people with SMI who can benefit and in terms of potential reductions in future criminal justice involvement. This R34 proposal brings together a team of nationally recognized experts to engage a deployment-focused approach to the development and preliminary testing of a new, scalable, and sustainable group-based, criminogenic-focused cognitive-behavioral therapy intervention developed specifically for use among people with SMI in community-based mental health settings. This proposed study includes three aims. The first aim will engage user-centered design methodologies to optimize the development of the new intervention. The second aim will conduct a small pilot randomized controlled trial to test the preliminary effectiveness of the new intervention. This pilot trial will engage an experimental therapeutics approach to examine how the new intervention engages the intended treatment targets and outcomes. The third aim will use qualitative research methods with key stakeholders to identify implementation strategies that both maximize and expediate the scalability and sustainability of the new intervention in community based mental health settings.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT CAREER GOAL: My long-term career goal is to lead an independent program of research that will leverage advancements in neuroimaging, discoveries from translational animal models, and robust clinical assessments to identify early pathology in brain development during critical periods in infancy when intervention can make the greatest impact. Ultimately, I aim to improve outcomes in children with neurodevelopmental disorders (NDDs), with a focus on Down Syndrome (DS), by contributing to work that will expand our understanding of early brain development and inform personalized, targeted interventions and non-invasive markers for treatment efficacy. RESEARCH PROJECT: Recent discoveries of the glymphatic system and meningeal lymphatic drainage have highlighted that cerebrospinal fluid (CSF) is critically important for maintaining brain health by clearing neuroinflammatory proteins (e.g., amyloid-beta, Aβ), whereas impaired CSF flow slows the clearance of Aβ and has been implicated in the pathogenesis of Alzheimer’s disease. Approximately 50% of children with DS will develop early-onset Alzheimer’s, occurring decades earlier than the general population. However, our understanding of CSF dynamics has been limited to studies in aging adults, whereas clinical studies in infants with NDDs (such as DS) are lacking. During infancy, the brain undergoes rapid growth and may be particularly vulnerable to CSF abnormalities, but there is a critical gap in understanding CSF physiology during this sensitive period and how it relates to the early brain development of NDDs. Given that children with DS are at a substantially greater risk for Alzheimer’s, there is an urgent need to study CSF dynamics in infants with DS to identify early, non-invasive biomarkers of disorder severity and progression and to guide personalized, targeted treatments. We aim to utilize non-invasive MRI methods in infants to evaluate three measures of CSF physiology (extra-axial CSF volume, perivascular space size, and CSF flow); their relationships to clinical manifestations of DS; and compared to related NDDs (autism and Fragile X syndrome). Specific Aims: (1) Elucidate trajectories of CSF dynamics in infants with DS and contrast with other NDDs to determine specificity; and (2) determine the relationships between CSF dynamics and (2a) neural and (2b) clinical features of DS and related NDDs. CAREER DEVELOPMENT: This K01 award will provide me with the necessary cross-disciplinary training in CSF imaging, CSF pathophysiology, and early neural and behavioral features of DS and NDDs to launch my independent career. Mentorship team includes clinical and preclinical experts in CSF abnormalities in NDDs, glymphatic system, and CSF and brain imaging in infants: Co-mentors: Drs. Mark Shen (Dept. of Psychiatry and Neuroscience, UNC) and Jeffrey lliff (Dept. of Neurology, University of Washington). Advisors: Drs. Joseph Piven (Psychiatry, UNC), Robert McKinstry (Radiology, Washington University), Juan Piantino (Pediatrics, Oregon Health and Science University), and Dr. Martin Styner (Computer Science, UNC).

NIH Research Projects · FY 2025 · 2022-09

Project summary One bottleneck to achieving therapeutically relevant cell concentrations at the infarct site after myocardial infarction is the poor cell engraftment and retention of vehicles at the target site. Here we propose the first-of-its- kind, tunable, replenishable scaffold of cells that allows for multiple drug delivery "waves" to address this gap. We hypothesize that cardioregenerative cell accumulation and retention in the infarcted myocardium will be enhanced by surface decorating them with proteins that cross-link the cells via layer-by-layer assembly into a scaffold. Unlike conventional delivery strategies, which do not allow the subsequent accumulation of cells or carriers after saturation of the target infarct, each dose of cells in our platform will serve as a capturing surface for the next dose of cells. This will dramatically amplify the targetable surface area for additional waves of cell attachment and will also allow the total therapeutic concentration to be adjusted based on the number of doses administered. To prevent premature cross-linking, we will use engineered proteins that form heterodimers but not homodimers. These studies are expected to result in a new class of carrier-linked network that will not only substantially enhance cellular accumulation, retention, and local drug release at the infarct site to maximize therapeutic efficacy but also allow for several cycles of drug replenishment or personalized dosing in a non- invasive manner. Here, we aim to evaluate the safety and therapeutic effectiveness of this approach and assess the effect of surface modification on stem cell function. Successful completion of the proposed studies will transform the treatment of patients suffering from myocardial infarction.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT / PROJECT SUMMARY Kinases, a class of proteins with more than 500 members in the human proteome, are important regulators of biological processes in health and disease. Kinases have proven to be excellent drug targets with more than 70 FDA approved medicines that target kinases. Despite this success, most kinases are understudied, and details of their functions are poorly understood. The NEK family of 11 kinases (NEK1 through NEK11) is a particularly understudied set of kinases that play roles in key biological processes like the cell cycle, ciliogenesis, and the DNA damage response (DDR), all with relevance to cancer and human health. These kinases have emerging links to numerous cancers, diabetes, inflammatory bowel disease, ciliopathies, and ALS. In this project we will use an efficient kinase systems-based approach to create an enabling suite of chemical probes, assays, reagents, and molecular tools to identify NEK family members that have key roles in cancer. These high-quality compounds and reagents we generate, which we will freely share, will allow scientists to build a deep understanding of the physiological and pathological roles members of the NEK family play. In Aim 1 we will create potent and selective inhibitors of each NEK using iterative medicinal chemistry and state of the art in cell target engagement assays. In a complementary effort for this aim we will also create inducible NEK knockdown cell lines. In Aim 2, using compounds and the NEK knockdown lines, we will evaluate the role and importance of each NEK in a suite of NEK and oncology-relevant cell health and cell biology signaling assays, measuring effects on proliferation, migration, the cell-cycle, DNA replication, and ciliogenesis. In Aim 3 we will experimentally determine the substrates of each NEK, locate the NEKs in broader kinase-dependent signaling pathways, and develop genetically targetable kinase activity reporters for tracking NEK activity within the endogenous cellular environment. Output from this project will include potent and selective NEK inhibitors, NEK family-wide assays, details on the impact of NEK inhibition and knockdown on key cancer processes, molecular tools, and NEK substrate and pathway information. Successful completion will provide a framework and the resources needed to validate individual NEKs as high quality, druggable targets for the treatment of cancer.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract During meiosis, crossing-over between homologs facilitates accurate chromosome segregation and prevents aneuploidy, which in turn forestalls miscarriages and chromosomal disorders such as Down syndrome. The regulation and placement of meiotic crossovers acts as a vital safeguard against age-associated meiotic defects and infertility, as the risk of non-disjunction (NDJ) increases with increasing maternal age. Meiotic crossovers (COs) are formed when programmed double-strand breaks (DSBs) are repaired through homologous recombination. However, only a subset of DSBs are repaired to form COs; the rest are repaired as non-crossovers (NCOs). Despite meiotic DSBs being distributed throughout the chromosome, CO placement is intricately regulated by three types of patterning phenomena. One of these, the centromere effect (CE), ensures the exclusion of COs in centromere-proximal regions and is crucial to the meiotic cell as centromere-proximal COs increase the risk of NDJ. Furthermore, increasing maternal age has been shown to weaken the CE, potentially explaining why NDJ incidence increases in older women. Although first observed in Drosophila in 1932, the mechanisms behind the CE remain unknown even today. The experiments proposed here aim to address this gap in knowledge regarding a vital cellular process that prevents mis-segregation events, especially in those with advanced maternal age. Recently, our lab showed that the CE is differentially established in the two classes of heterochromatin found at the Drosophila pericentromere. In the highly repetitive alpha heterochromatin immediately adjacent to the centromere, a complete exclusion of COs is observed, while the less repetitive beta heterochromatin adjacent to proximal euchromatin shows a distance dependent CO suppression. I will build on these results by investigating the mechanisms of how the CE is established in these two classes of pericentric heterochromatin. A prominent question regarding CE mechanisms centers around how pericentromeric heterochromatin and the centromere itself contribute to the CE. The few studies that have addressed pericentric crossing-over in the past century have attempted to establish one as more important than the other in Drosophila but failed to arrive at a consensus. Thus, pericentric heterochromatin has been considered everything from an active participant in CO reduction in adjacent intervals to nothing more than a passive spacer between euchromatin and the centromere. Through the experiments outlined in this proposal, I will ask how pericentric heterochromatin and the centromere contribute to the CE independently of each other, particularly focusing on highly repetitive alpha heterochromatin. Investigating the role of alpha heterochromatin as separate from that of pericentric heterochromatin as a whole in manifesting the CE is a novel area of research within the broader question of how the CE is established. In summary, this proposal will increase our understanding of the mechanisms that safeguard against age- related aneuploidy and infertility through shedding light on meiotic crossover patterning.

NIH Research Projects · FY 2025 · 2022-08

Modified Project Summary/Abstract Section This K24 Midcareer Investigator Award in Patient Oriented Research (POR) will leverage my existing POR program, enhance my professional development and that of my trainees, and support my ongoing efforts to provide essential mentoring to early stage clinical investigators in POR in the rheumatic and musculoskeletal diseases (RMDs), with a focus on osteoarthritis (OA). Despite being a highly prevalent and serious disease, OA lacks effective therapies and remains a large burden on individuals and the health care system. We have published extensively on social determinants of health in various aspects of OA based on our experience with two population-based cohorts, the Johnston County OA Project (JoCoOA) and the Johnston County Health Study (JoCoHS). My POR career has focused on aspects of OA epidemiology, imaging, and biomarkers, with a more recent emphasis on identifying phenotypes using machine learning and precision medicine to improve studies and management of this chronic disease. It is essential to understand the role of artificial intelligence, including machine learning, in either improving or perpetuating barriers to care such as those known to exist in OA. Such advanced approaches, if applied appropriately, will likely provide new insights on phenotypes of OA, and inform therapeutics across populations with the objective of improving health for all. Further, the intersection of POR, population health, and machine learning is of great interest to trainees and an area of critical need for training future clinician scientists in RMDs, which is the goal of the K24 mechanism. We will leverage existing data from the JoCoOA, a 30-year longitudinal study of over 4000 Black and White men and women aged 45 and older, and the new JoCoHS, an actively enrolling cohort (2019-, n~1500) that includes individuals who are 35-70 years of age and who identify as Hispanic, Black, or White. We will utilize these rich data sources and our extensive institutional resources to address two specific aims around subgroups of pain and symptoms and their associations with structure and function, with a focus on differences in outcomes. In Aim 1, we will work to validate and optimize a deep learning algorithm in these representative cohorts to better define the association between radiographic features and the pain experience in knee OA. In Aim 2, we will characterize symptomatic phenotypes in the JoCoHS cohort (which has more extensive available symptomatic data), and associate these with the radiographic information gleaned in Aim 1 as well as validated assessments of function and physical activity, to identify subgroups that could most benefit from targeted interventions. This K24 project will provide crucial protected time and resources to promote my professional development and enhance my mentoring capabilities while simultaneously providing a springboard for trainees to 1) develop their research skills in the context of individualized projects, 2) develop critical skills in big data analytics and machine learning-based methodology, and 3) answer key POR questions in RMDs and OA.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT By the age of 5, children in low and middle-income countries (LMICs) are exposed to nearly five times more antibiotics than high-income country children. Although improved access to antibiotics has been a major driver of mortality declines, most antibiotics administered to children are clinically unnecessary. Excessive use can lead to adverse events, drug toxicity, and harm the gut microbiota and immune system. It also contributes to antimicrobial resistance (AMR), the costs of which are disproportionately borne by children in LMICs. Although widespread clinically unnecessary use of antibiotics in LMICs is well-documented, substantial knowledge gaps remain regarding the drivers of overuse among children and how these are linked to the dynamics of resistance and disease. This knowledge is required to design policies and interventions that appropriately balance access and overuse. This K01 Award proposal focuses on identifying incentives that caregivers and providers face to treat children with antibiotics and how these are related to the development of resistance. My career goal is to become independent scholar working at the intersection of economics and infectious disease epidemiology with a focus on research to inform AMR policies in LMICs. The proposed training activities build on my background as an economist and experience conducting population-based experimental research with further training in infectious disease epidemiology, the biomedical underpinnings of antimicrobial resistance, machine learning techniques, and agent-based modeling of infectious disease and social systems. Aligned with my training goals, my research program aims to integrate concepts from economics and infectious disease epidemiology and to use state-of-the-art machine learning approaches to examine the complex relationship between factors driving demand for antibiotics, disease, and the development of resistance. To do so, I will draw on existing micro-level data from a survey of clinicians and households across 360 rural villages in southwest China as well as new experimental data on the prescription practices of clinicians and pharmacists in the same area. My specific research aims are 1) to experimentally evaluate the prescribing practices of clinicians and pharmacists for pediatric diarrhea cases; 2) to estimate the influence of clinician advice on antibiotic use in children, and how this varies with patient, clinician, and community characteristics; and 3) to develop an agent-based model of health-seeking behavior, antibiotic use, and bacterial resistance for pediatric diarrhea cases in rural China and use this model to conduct counterfactual simulations to prioritize interventions for future study. My mentoring team has specialized training in infectious disease epidemiology, the epidemiology of antimicrobial resistance, machine learning, and agent-based modeling as well as experience leading interdisciplinary teams. This research will generate new insights that can inform policies to better balance access to antibiotics and overuse. The training and research proposed in this K01 award will support the development of future R-level proposals to study the design of AMR policies in LMICs.

NIH Research Projects · FY 2024 · 2022-08

ABSTRACT Throughout the SARS-CoV-2 pandemic, explosive transmission of infection has occurred in jails and prisons. The spillover of infection between jails, prisons, and surrounding communities is fueled by high churn of the incarcerated population and daily cycling of correctional staff between facility and community. Failure to control transmission in correctional settings has resulted in significant COVID-19 morbidity and mortality and contributed to continuing racial health disparities. Despite emerging evidence of disproportionate spread of SARS-CoV-2 infection arising from jails and prisons, few studies have examined transmission dynamics in correctional settings or evaluated interventions to reduce COVID-19 morbidity and mortality. The objective of this Mentored Research Scientist Development Award (K01) is to equip Dr. Levintow with essential training in criminal justice research and infectious disease modeling to investigate the dynamic processes that drive SARS-CoV-2 transmission and advance intervention efforts to prevent COVID-19 morbidity and mortality, focusing on jails in North Carolina (NC). Specifically, a K01 award will provide substantive knowledge of the criminal justice system and infectious diseases in that context (Training Aim 1), methodological expertise in mathematical modeling of infectious disease dynamics (Training Aim 2), and new skills in linking system-level policy and interventions to epidemiological outcomes (Training Aim 3). Leveraging this training, Dr. Levintow will apply new expertise about infectious diseases in the criminal justice system to assess the extent to which population movement, COVID-19 testing, and COVID-19 cases in NC jails are predicted by jail characteristics (Research Aim 1). Guided by intensive training in the development, calibration, and analysis of mathematical models, Dr. Levintow will estimate SARS-CoV-2 incidence in NC jails and corresponding excess infection relative to surrounding communities (Research Aim 2). With new knowledge of existing and proposed interventions in jails – coupled with new skills to rigorously assess their effects – Dr. Levintow will evaluate the impact of those interventions on COVID-19 morbidity and mortality (Research Aim 3). Study findings will inform prioritization of interventions with greatest impact for the highly vulnerable populations in NC jails and their surrounding communities across the state. This study will benefit from the institutional resources of the University of North Carolina at Chapel Hill and will be guided by a mentoring team of leading experts with extensive experience examining infectious disease in the criminal justice system and conducting modeling analyses to inform interventions on spread of infection. This K01 award will provide both the training needed to accomplish the proposed research and the support necessary to launch Dr. Levintow's independence as an infectious disease epidemiologist, laying the groundwork for R01-level research expanded to the national level and to other infectious diseases with potential for explosive outbreaks in jails and prisons.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Primary cilium, a microtubule based antennae-like organelle, is present in every cortical neuron, and when defective, leads to ciliopathies. A distinguishing feature of ciliopathies is defective neural circuit formation and function. Disrupted construction and function of neural circuits in ciliopathies lead to disorders such as autism, intellectual disabilities, mood disorders, obesity, and epilepsy, thus implying a role for primary cilia in neuronal function and circuit dynamics. Primary cilia signaling may serve as a non­synaptic signaling mechanism through which environmental signals can shape and refine neuronal circuits in health and disease. Nonetheless, how primary cilia signaling sculpts neuronal circuit dynamics and whether primary cilia can be co-opted as a therapeutic conduit to mend neural circuit malfunctions remain enigmatic. We aim to resolve this challenge by defining the signaling mechanisms that are utilized by primary cilia to enable appropriate neuronal functions necessary for the emergence and maintenance of functional neural circuits in the brain. We will make this goal attainable by leveraging the latest advances in optogenetic and chemogenetic interrogation of signaling emanating from primary cilia, mapping neuronal ciliary receptome, profiling ciliary connectome within human cerebral cortical circuitry, live imaging cilia driven neuronal activity and transcriptional changes during neural circuit plasticity in living animals, and modeling human ciliopathies in brain organoids with the aim of rescuing circuit malfunctions using primary cilia as a tool. Collectively, this work will reveal how primary cilia activity is transformed into changes in neuronal circuit function and the pathways that must be successfully engaged to harness this insight in the service of ameliorating neural circuit disorders. These outcomes will offer a transformative opportunity to define new cellular principles of neural circuit formation and function and will open up new therapeutic avenues of neural circuit correction.

NIH Research Projects · FY 2025 · 2022-08

Project Summary/Abstract More than 75% of persons living with Alzheimer's disease and related dementias who reach 80 years of age require residential long-term care, which is increasingly provided in assisted living (AL). Across the country, almost 29,000 AL communities with more than 996,000 beds have become the primary residential care provider for persons with dementia: 90% of AL residents have cognitive impairment and 42% have recorded moderate or severe dementia, with actual rates being higher. AL provides supportive but not nursing services; consequently, virtually all care is provided by direct care workers (nursing assistants and personal care aides). Unfortunately, direct care workers are undervalued and undertrained, leading to poor care, workplace injury, dissatisfaction, and high turnover. AL is state-regulated, and only 17 states stipulate minimum training hours (some being as low as one hour), meaning that two-thirds of states are silent on training. Fewer than 40% of staff have education beyond high school, and so it is not surprising that a minority report sufficient knowledge to care for persons with dementia. In striving to fill this gap, training for direct care workers must be accessible and have efficacy in benefitting the staff, organization, and persons with dementia. Online training is an especially promising option due to its low cost, wide availability, and potential for self- pacing, automated skills tests, and certification. The Alzheimer's Association is the national leader in dementia care training, and in 2021 developed essentiALZTM (pronounced “essentials”), an online program teaching evidence-based, person-centered care, which can be accessed from a computer, tablet, or mobile device. Already more than 1,500 staff have essentiALZ certification, but as is true of the majority of training programs, evidence as to its ability to improve care and outcomes is lacking. It is possible that essentiALZ is effective in changing care and outcomes, but it may also be that additional supports are necessary to do so. A timely model of support is Project ECHO, which has flooded the field of long-term care as a proven way to provide expert guidance and peer support via a remote, online approach. Adding ECHO to online dementia training might provide a necessary boost to achieve care change and improved outcomes. The proposed project responds to the NIA Notice of Special Interest that calls for strengthening the workforce through enhancing and supporting skills training. It will conduct a hybrid implementation/effectiveness cluster- randomized trial in 126 AL communities across six states, comparing essentiALZ alone, essentiALZ + ECHO enhancement, and a waitlist control. Outcomes grounded in the RE-AIM model and the Kirkpatrick training effectiveness model will be examined over six months, comparing the arms in terms of (1) implementation and (2) effectiveness, and (3) examining the extent to which implementation and effectiveness differ based on characteristics of the AL community, staff, residents, and family members. Results will inform next steps in dementia care training for the AL and broader long-term care workforce.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY/ABSTRACT Cardiovascular disease (CVD) disparities are increasingly attributed to the distinct social conditions in which various social groups age. Socioeconomic and psychosocial factors are known to cause chronic stress that may increase risk of CVD, but the mechanisms are not fully clear. It is therefore important to identify factors that mediate or moderate the effects of social stressors on CVD. Geroscientific theory reconceptualizes chronic disease as a product of sustained dysfunctional aging, offering additional avenues for understanding how the social environment is internalized to influence health. Work emerging from this literature has identified epigenetic dimensions of aging such as DNA methylation which appear particularly responsive to psychosocial stressors. Epigenetic “clocks”, or measures of DNA methylation-based predictors of chronological age, health, and mortality may serve as useful markers that can provide important missing information on the relationships between social stressors, aging, and CVD. Furthermore, psychological factors may also play an integral role in these pathways. Self-concept is a determinant of a number of identity-based psychosocial characteristics associated with both epigenetic changes and CVD. These include dispositional traits such as optimism and negative affect, as well as beliefs about self-worth and subjective social status. Importantly, these factors are also known to influence the way individuals perceive and cope with stress. Antecedent to the psychosocial traits which influence stressor appraisal and stress-responsive behavior, self-concept may therefore serve as a viable point of intervening on the subjective identity characteristics which catalyze premature and dysfunctional aging in the context of social disadvantage and predispose individuals to CVD. Incorporating a geroscientific perspective, the applicant’s Identity Vitality-Pathology (IVP) model theorizes that certain self-concepts render individuals particularly vulnerable or resilient to the health effects of social stressors by promoting or preventing premature aging. The proposed IVP scale (IVPS) aims to measure these novel identity characteristics hypothesized to predict aging-related psychosocial factors and cardiovascular health. The specific objectives of the proposed study are to: 1) examine whether psychosocial risk and resilience factors predict biological age as measured by epigenetic clocks using data combined from the Jackson Heart Study and the Atherosclerosis Risk In Communities cohorts; and 2) use preliminary qualitative data to develop and finalize items for use in the IVPS. R00 phase aims include examining the psychometric properties of the IVPS and assessing whether self-concept as measured by the IVPS predicts CVD-related psychosocial traits and biological aging. Ultimately, this work has the potential to identify additional targets for addressing racial and gender disparities in healthy aging and CVD, particularly ways of favorably influencing epigenetic changes associated with healthy aging.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Chromatin remodeling complexes play a critical role in regulating gene expression, differentiation, and development. The SWI/SNF chromatin remodeling complex is mutated in ~25% of all human tumors. The 12-15 positions within SWI/SNF are filled from ~30 unique subunits. The combinatorial assembly of these genes can yield 1000-2000 biochemically distinct complexes . While often studied as a single complex, these biochemically distinct forms of SWI/SNF have different functions in gene regulation.Understanding the mechanisms that regulate SWI/SNF function, and that are therefore disrupted in SWI/SNF mutant tumors, is of critical importance for developing new therapies. We propose that post-translational modifications of SWI/SNF and interactions between SWI/SNF and RNA modulate function of the complex. Understanding how these mechanisms contribute to how different SWI/SNF complexes are defined is the focus of my lab. The key questions we will address in this project are 1.) Do variant SWI/SNF complexes have different functions? 2.) What regulates different activities of biochemically distinct complexes? 3.) How are changes to the composition and the targeting of complexes regulated? Our lab combines computational, molecular, and biochemical analyses to answer these questions and develop deeper insight into the mechanisms of SWI/SNF activity. Over the next five years, we will investigate mechanisms that combine to regulate the composition and function of SWI/SNF and determine how different functional outcomes are mediated by distinct SWI/SNF complexes. Given the critical role of SWI/SNF in development and disease, understanding how the complex is regulated to mediate different outcomes is critical for developing new approaches to targeting cancer and other diseases.

NIH Research Projects · FY 2025 · 2022-08

OVERALL ABSTRACT The mucus clearance system constitutes the primary airway host defense system against inhaled infectious agents and toxins. However, despite more than two centuries of research into the nature of the mucus clearance system, surprising gaps in our knowledge of fundamental aspects of this system persist. Filling in these gaps is important for improving public health strategies to combat respiratory infectious diseases. Filling in these gaps is also important for elucidating the pathogenesis of and developing therapies for chronic pulmonary diseases, including COPD, asthma, NCFB, and rare genetic diseases (CF, PCD), which are by definition characterized by mucus accumulation in the lung. This PPG proposes to investigate fundamental, but poorly understood, aspects of the mucus clearance system that must be quantitated to understand mucus function in health and dysfunction in disease. Each PPG project has two specific aims focused on basic mucin function and one focused on translational aspects of mucin pathobiology. Project 1 (“Mucin Structure and Associations in Respiratory Mucus”, Michael Rubinstein, PhD, PI) will investigate fundamental aspects of the organization of mucins in solution and within the mucus layer. These studies will be complemented by studies of the addition of “abnormal polymers”, e.g., DNA, to mucus solutions. Project 2 (“Why are mucins so gigantic and is it safe/effective to sever them therapeutically?”, Richard C. Boucher, MD, PI) will focus on the fundamental question as to why human airway mucin polymers are of such enormous size (300 MDa, Rg 250 nm) and characterize the ratio of efficacy (chain length reduction) vs risk (off-target chain unwinding) required for the development of mucolytics for lung disease. Project 3 (“Membrane-bound mucins on the airway surface ensure efficient mucus clearance and lung health”, Brian Button, PhD, PI) will study the relationships between cilia, PCL, and the mucus layer required for transport, focusing on a novel hydraulic “pushing” vs classic “clawing” mechanisms. In addition, barrier functions of PCL and regulation thereof will be studied. Project 4 (“Biophysical and structural characterization of airway submucosal gland mucus in health and cystic fibrosis”, Ronit Freeman, PhD, PI) will focus on a novel attribute of submucosal gland (SMG) mucus, a strand/bundle insoluble component, and how strands/bundles contribute to SMG mucus function in health and disease. Three cores support the PPG: 1) Core A, the Administrative/Biostatistical Core, Multi-PIs Richard C. Boucher, MD, and Michael Rubinstein, PhD, supplies project management and statistical support for the PPG; 2) Core B, the Mucus/Mucin Analytics Core, PI Mehmet Kesimer, PhD, provides quality control of all mucin reagents for the PPG and novel biochemical/biophysical measurements; and 3) Core C, the Imaging Core, PI Camille Ehre, PhD, provides electron microscopic, molecular, and morphologic analyses to the projects. The overall goals of the PPG are to elucidate the structure and function of mucus in health, how these characteristics are degraded in disease, and identify strategies for development of novel therapeutic agents to treat muco-obstructive diseases.

NIH Research Projects · FY 2025 · 2022-08

Project Summary The central hypothesis of this work is that increased intestinal permeability (IP), either directly, or via related comorbidities, promotes the development and worsening of multi-joint osteoarthritis (MJOA). Multiple joint osteoarthritis (MJOA; referring to OA in more than one joint site within an individual) is common but understudied. MJOA is progressive, and as whole-body burden of OA increases, associated pain and disability increases, and treatments are less successful. Despite the significant societal impact of MJOA, most OA research remains focused on individual joints. There is an urgent need to understand the factors that promote progression and worsening of MJOA. To address our hypothesis, our group has access to a large, longitudinal cohort of human patients, and, uniquely, access to the naturally occurring MJOA model in pet dogs. There are no rodent models of MJOA, but dogs with naturally occurring MJOA have similar disease manifestations with more rapid progression compared with humans, making pet dogs an ideal model in which to explore underlying mechanisms of MJOA and potential therapies. We have shown that inflammatory mediators are related to overall burden of OA; these and other risk factors may at least partly derive from the gut microbiome via increased intestinal permeability (IP; “leaky gut”). We have evidence that lipopolysaccharide (LPS) and LPS-binding protein (LBP, reflecting increased IP and increased exposure to microbial products), promote OA. Additionally, serum LPS in humans (and serum LBP in dogs) is positively associated with the number of joints affected by MJOA. To further elucidate the role of IP as a mechanism in MJOA, the proposed work will leverage human and dog studies: The JoCoOA, a longitudinal cohort of over 4000 Black and White men and women aged 45 and older; The Johnston County Health Study (JoCoHS), an actively enrolling cohort (2019-, n~2000) including younger (35-70 years) and Hispanic individuals; and a large cohort of readily accessible naturally occurring MJOA in pet dogs. Data from all three cohorts will be used to address the following three aims. In Aim 1, we will determine cross-sectional associations between altered IP, systemic inflammation, and radiographic and symptomatic MJOA in humans and pet dogs. Aim 2 will allow identification of biomarkers predictive of development and worsening of MJOA and determine longitudinal associations with markers of systemic inflammation and IP among JoCoOA participants and dogs. In Aim 3, we will test the effects of a prebiotic on IP, the microbiome and MJOA symptoms by randomizing 70 dogs with MJOA (from Aim 1) to receive either a fructooligosaccharide supplement or placebo for 3 months followed by re-characterization of biomarkers of inflammation and IP. These studies will both verify the association between increased IP and MJOA and robustly define biomarkers predictive of development and worsening MJOA, laying the groundwork for mechanistic studies to understand how increased IP promotes MJOA and to identify therapeutic targets, as well as provide means to identify at-risk individuals for preemptive management.

NIH Research Projects · FY 2024 · 2022-08

Project Summary/Abstract: Transgender (trans) women across the globe experience a disproportionate burden of HIV with an estimated global HIV prevalence of 19%. Trans women living with HIV also have worse HIV care and treatment outcomes than cisgender adults, including lower rates of viral suppression. Multilevel responses are needed to improve HIV outcomes among trans women by simultaneously and synergistically addressing individual, provider, and community dynamics. Trans women experience intersectional stigma due to the frequent association between trans identity, HIV, and sex work. These intersecting forms of stigma include internalized stigma in the form of applying negative beliefs and enacted stigma in the form of exclusion and discrimination. Developing social cohesion can be a critical first step in community processes to address stigma. We adapted a multilevel intervention, Abriendo Puertas (Opening Doors), including individual counseling, peer navigation, and community mobilization for trans women living with HIV in the Dominican Republic using an iterative consultation process. We assessed feasibility and initial effects on HIV care and treatment behaviors with 30 trans women living with HIV (no control group) and documented positive trends in ART use (70% to 85%, p=0.03), missed care appointments (35% to 20%, p=0.39) and ART adherence (86% to 96%, p=0.50). Participants emphasized that trusting intervention staff and being treated with respect in individual sessions allowed them to improve self-esteem. Limited trust and cohesion among trans women, however, inhibited more extensive engagement with peer navigation and community activities. In response, we identified two key modifications to strengthen and further tailor the intervention for trans women living with HIV: 1) integrate more gender affirming content, including with providers and 2) focus on building trust among trans women through sequential implementation of individual and then community components. The purpose of the proposed study is to conduct a pilot randomized trial of the GAP model, or Gender-affirming Abriendo Puertas. In Aim 1, we will assess preliminary efficacy of the GAP intervention on viral suppression among trans women randomized to the intervention compared to those randomized to control. We will randomly assign trans women living with HIV to the GAP intervention (n=60) (individual counseling, peer navigation, provider training, and community support building) or control group (n=60). We will conduct baseline, 6, and 12-month surveys and viral load assessments to assess differences across study arms. In Aim 2, we will examine pathways of influence (e.g. decreased stigma, increased cohesion) and experiences with the intervention to identify specific areas for improvement and scale up. We will conduct longitudinal qualitative interviews at baseline, 6, and 12 months with 20 intervention participants and, together with surveys, assess how GAP participation affects pathways between stigma, cohesion, and HIV outcomes. We will also elicit experiences and recommendations from providers and intervention staff in focus groups at 6 (n=2) and 12 months (n=2).

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Flaviviruses such as Powassan virus (POWV), West Nile virus (WNV), and Japanese encephalitis virus (JEV) are transmitted by ticks and mosquitoes. The outcomes of flavivirus infection are heterogeneous, with only a subset progressing to neuroinvasive disease (e.g. encephalitis, meningitis, or paralysis). We hypothesize that host genetic variation, particularly in antiviral response genes, contributes to differential disease outcome following flavivirus infection. The Collaborative Cross (CC) panel of recombinant inbred mice provides an ideal system to discover novel mechanisms of immune-mediated control of flavivirus pathogenesis because these mice exhibit an expanded range of immune phenotypes on reproducible genetic backgrounds. We infected 17 CC lines with POWV and identified multiple highly susceptible lines, including CC071 and CC015, and a single resistant line, CC045. Most phenotypes were concordant among POWV, WNV, and JEV, but some lines exhibited virus-specific resistance, implying that there are both virus-specific and pan-flavivirus mechanisms that control resistance to neuroinvasive flaviviruses. We propose to use the CC to determine the viral and immunologic features of POWV pathogenesis and to identify host genes that contribute to POWV resistance. Aim 1: Define viral and immunologic features of POWV pathogenesis in CC mice. We found that CC045 mice exhibited equivalent viremia but lower brain viral loads compared to CC071 mice, suggesting that POWV resistance may result from reduced neuroinvasion. We will assess brain viral loads and infiltrating leukocytes in susceptible and resistant CC lines following POWV infection. We will assess blood-brain barrier permeability at baseline and in response to viral infection and inflammatory stimuli. We will generate primary cells from susceptible and resistant CC lines and measure replication of POWV and other flaviviruses. Aim 2: Map quantitative trait loci and evaluate antiviral activity of host factors associated with POWV resistance. To identify polymorphic host genes that determine the outcome of POWV infection, we generated two F2 crosses of susceptible and resistant lines (CC071 x CC045 and CC015 x CC045) and evaluated lethality in ~300 F2 mice per cross following POWV infection, as well as CNS viral loads in ~120 F2 mice. We will map QTL associated with POWV resistance in both crosses and investigate candidate genes under significant QTL. Aim 3: Distinguish pan-flavivirus and virus-specific restriction factors in CC mice. We will infect additional CC lines with JEV to identify lines that are differentially resistant to JEV compared to POWV or WNV. We will evaluate brain viral loads and infiltrating leukocytes following JEV infection. We will generate F2 progeny of susceptible and resistant lines and map QTL associated with JEV resistance. The proposed studies will provide insight into the pathogenic mechanisms of POWV and reveal polymorphic host immune mechanisms that impact susceptibility to flavivirus neuroinvasive disease. This work will provide the foundation for future investigations of novel immune factors that control flavivirus pathogenesis.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Obesity is associated with increased risk of developing and succumbing to endometrial cancer (EC), with ~60% of EC patients being obese. Since programmed death protein-1 (PD-1) and its ligand, programmed death-ligand 1 (PD-L1), are highly expressed in ECs, immuno-oncologic inhibitors for these two targets hold great promise for the treatment of obesity-driven EC, especially as ~25% of ECs have microsatellite instability, a known biomarker for PD-1/PD-L1 inhibitor response. Atezolizumab is one such anti-PD-L1 monoclonal antibody (mAb). Our studies suggest the obesity-triggered pro-inflammatory uterine tumor milieu increases PD-L1/2, culminating in enhanced susceptibility to atezolizumab. The efficacy of atezolizumab in EC may be enhanced via ONC201, a small molecular selective dopamine receptor 2 antagonist that upregulates Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand and activates T and natural killer (NK) cells. Thus, ONC201 is a logical therapeutic partner for atezolizumab. However, the use of atezolizumab + ONC201 in EC is complicated by obesity. Compared to small molecule drugs, mAbs are cleared via cells of the mononuclear phagocyte system (MPS; monocytes) which is part of the innate immune system. MPS cells serve as a natural mechanism of uptake and clearance for mAbs via their Fc- gamma-receptors (FcɣRs). Our studies show MPS mediators, number of FcɣRs and function in blood are highly variable and associated with the high and clinically relevant variability in the pharmacokinetics/pharmacodynamics (PK/PD) of mAbs. Obese patients have higher and more variable MPS mediators, FcɣRs and function, resulting in lower exposures of mAbs. Thus, evaluating effects of obesity on the PK/PD of atezolizumab and ONC201 in EC patients is clinically relevant and critically important. Our hypothesis is that atezolizumab + ONC201 will be safe in EC; however, despite obese EC patients having higher expression of PD-L1 and a pro-inflammatory tumor milieu, they will have lower atezolizumab plasma levels, than the non- obese at the same dose due to increased phagocytic clearance, necessitating higher dosing in the obese to achieve serum levels comparable to those of the non-obese patients. We propose a phase 1 clinical trial of the novel combination of atezolizumab + ONC201 using an innovative design of parallel cohorts of obese and non-obese patients with metastatic and recurrent EC. Our primary objective is to evaluate the safety and tolerability of the dual regimen of atezolizumab and ONC201 in obese and non-obese EC patients to find the maximal tolerable dose combination to then advance for both cohorts in a future phase 2 study. In order to comprehensively delineate the impact of obesity on the combination of atezolizumab + ONC201, we will compare between the obese and non-obese EC groups the PK/PD of both agents, biomarkers reflective of the immune-oncology and of the MPS clearance of both agents, and their inflammatory metabolic signatures.

NIH Research Projects · FY 2025 · 2022-08

1. Summary Atherosclerosis is a leading cause of morbidity and mortality world-wide. It is clear that vascular smooth muscle cells (SMCs) play a critical role in plaque progression and stability, but many questions remain in regard to the source, fate, and function of the phenotypically modulated SMCs within the protective fibrous cap and necrotic core. Defects in SMC function that lead to hypertension (HTN) and increased vascular stiffness also affect plaque formation and progression by altering mechanical signaling within the vessel wall. Although these physiologic parameters are major independent cardiovascular risk factors, we know surprisingly little about their development, their inter-relationship, or the mechanisms by which they promote atherosclerosis. We have previously shown that the SMC-selective Rho-specific GAP, GRAF3, reduces blood pressure in mice and humans by limiting RhoA-dependent SMC contractility in resistance arterioles and went on to identify rs604723 as the causal variant within the BP-associated locus in this gene. Our more recent data indicate that GRAF signaling is atheroprotective and inhibits the expression of the contractile and extracellular matrix genes that drive vascular stiffness, and the pro-inflammatory and pro-calcification gene programs that contribute to atherosclerotic plaque development and rupture. The overall goals of this proposal are to assess the role of GRAF3 genotype on cardiovascular outcomes in a large and diverse cardiovascular disease patient population, to directly measure the contribution of GRAF signaling to atherosclerosis and vascular stiffening, and to use our understanding of GRAF intra-molecular interactions to identify GRAF-activating compounds that could be useful for treating HTN and atherosclerotic disease.

NIH Research Projects · FY 2025 · 2022-08

K. Gordon, UNC R35 Abstract The specification and robust maintenance of the germ line stem cells and their differentiated descendants that form gametes are essential to the success of an organismal lineage. Stem cell niches are primary regulators of stem cell fate, which is characterized by continuous self-renewal with the capability of differentiation. Therefore the structure of stem cell niches—which often have extensive cellular contacts with their supported stem cell populations that must be broken by daughter cells that go on to differentiate—appear crucial for the regulation of the cell fate decision. Such contacts are shared in a variety of stem cell niches, but studies relating niche structure to function lag behind genetic analysis of stem cell regulation. Stem cell niches lie deep in tissues, making them difficult to observe, and stem cell divisions are stochastic. This proposal avoids these challenges by studying the germ line stem cell niche of C. elegans, an animal that is suited to multiplex in vivo imaging. The overall goal for the research program is to discover how undifferentiated germ stem cells receive cues from their micro-environment to stimulate proliferation and the cell fate switch to gamete differentiation, and how the dynamic cell structures supporting germ stem cells form. Conserved proliferation and cell growth pathways are relevant to human health. Pursuit of these questions will lead to insights that, like other findings made in this canonical stem cell niche model, may be broadly applicable to other stem cell systems. Overview of research and goals for the next five years: The Gordon Lab opened at UNC just before the pandemic began. It pursues key questions like: How do the stem cell niche and gonad sheath cells grow and internally partition themselves to interact appropriately with germ cells at different steps of the differentiation program? Both the niche cell and sheath undergo substantial growth and transformation during development. Both somatic cells will be genetically manipulated to determine if effects on germ cell proliferation are mediated by growth and partitioning of these regulatory cells. Cell-specific RNAi strains will be used to screen for genes resulting in diminished Sh1 growth and germ cell proliferation. What molecular mechanisms underlie oriented cell divisions at the niche boundary and asymmetrical cell fate acquisition? Spindle orientation has been studied in great detail in early C. elegans embryos, but it was not known that divisions in proliferating germ cells were oriented. Adhesion between the niche and germ cells was a neglected area of inquiry until recent findings implicated cadherin complexes—which orient spindles in the embryo—in the dramatic wrapping behavior of the niche around the stem cells. How does the stem cell niche mature from larvae to adults? RNA- seq of niche cells across the larva/adult transition will identify candidate genes that are correlated with the transition from a migratory niche during gonad growth to a stationary, mature adult niche. These and known genes will be investigated by tagging and loss of function analysis in live-imaging studies of the transition. Together this work will address arising and long-standing questions about a classical stem cell model system.

NIH Research Projects · FY 2024 · 2022-08

Project Summary/Abstract Acute Myeloid Leukemia (AML) is a prevalent and deadly cancer, with a predicted 20,000 new cases and 11,000 deaths from AML this year in the US. A common treatment for AML is allogeneic stem cell transplantation (alloSCT). Though potentially curative, approximately half of all patients that receive alloSCT eventually relapse and die of their disease. In alloSCT, a patient’s immune system is suppressed and bone marrow hematopoiesis ablated then reconstituted with donor hematopoietic stem cells and leukocytes. Donor T cells are able to recognize peptides derived from recipient genetic polymorphisms as foreign and destroy the cells presenting them. These peptides that differ between donor and recipient are called minor histocompatibility antigens (mHAs). If mHAs are presented on AML cells, donor T cells will kill the AML cells in what is called the graft versus leukemia (GvL) effect. However, donor T cells also target peptides presented on healthy recipient tissues causing a life-threatening side effect called graft versus host disease (GvHD). Separating GvL from GvHD is a pivotal problem in alloSCT biology. T cell targeting of mHAs that are derived from proteins only expressed in the blood is viewed as a way to augment GvL without boosting GvHD. We hypothesize that mHAs mismatched between donor and recipient are a key determinant of alloSCT outcome for AML. In this work, I investigate mHA targeting in alloSCT for AML from two perspectives: mHA and T cell. My work will elucidate the role of mHAs in alloSCT, enrich clinical assessment of AML prognosis, and identify new mHAs for future therapeutic targeting. The training in computational and wet lab immunology included here will forward my goal of becoming an independently funded physician-scientist leading a research lab in leukemia immunobiology and caring for leukemia patients. We have computationally predicted mHAs using a large dataset of over 3000 alloSCT patients. In our mHA- focused Aim 1, I will validate our predicted mHAs using mass spectrometry to identify whether they are presented on the cell surface of AML cell lines and therefore can serve as targets for T cells. Using the mHAs I validate, I will statistically define peptide features that predict presentation on the cell surface by HLA, informing future mHA identification work. I will also assess associations between validated mHAs and clinical outcome after alloSCT. I will analyze mHAs by population frequency in all ethnic groups within the US, with the goal of identifying a minimal set of mHAs that cover the majority of AML patients of all ethnicities. In our T cell-focused Aim 2, we will assess whether T cells specific for mHAs are exhausted in patients experiencing relapse after alloSCT. We predict that efficacy of these T cells predict success of alloSCT, and that exhaustion of these cells will accompany relapse. We will investigate presentation of exhaustion markers and release of proinflammatory cytokines from mHA-specific T cells from alloSCT patients before and after relapse.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Psychiatric disorders are highly prevalent following kidney transplantation (KT) and significantly reduce the survival and quality of life of KT recipients. KT is a severely limited resource, and extensive efforts are made by transplant programs to anticipate post-KT co-morbidities. However, major depressive disorder (MDD) and post- traumatic stress disorder (PTSD) both have ~20% point prevalence in KT recipients. The overarching aim of this proposal is to systematically investigate genomic, clinical, and psychosocial contributors to MDD and PTSD following KT. The specific aims are: 1) generate a retrospective electronic health record-based cohort of KT recipients to characterize biopsychosocial risk factors for psychiatric disorders post-KT; 2) establish a prospective cohort of KT candidates to follow trajectories of psychiatric symptoms and risk factors; and 3) develop a predictive model for psychiatric disorders in KT recipients. Kidney disease disproportionately impacts traditional minorities, and most KT recipients are not of European ancestry. When calculating genetic risk score for KT recipients, I will leverage new and diverse training sets and a growing toolkit of genomic methods to appropriately estimate genetic risk across ancestry. I anticipate that this work will improve our ability to understand and mitigate risk of psychiatric disorders for kidney disease patients more broadly and will build on efforts to apply genomic data to patients with non-European ancestry. Dr. Nash is a physician-scientist deeply committed to her primary career goal of improving the mental health of severely ill patients, with an emphasis on those awaiting or living with a solid organ transplant. Her long-term research goal is to build clinical registries with genomic data to elucidate risk factors and etiology of psychiatric disorders in solid organ transplant recipients that will inform the future ideal of personalized, targeted, and pre- emptive treatment for those at risk for psychiatric co-morbidities. Dr. Nash has a PhD in biological chemistry with a strong scientific background, is a fellowship-trained Psychiatrist, and now requires training in clinical research and psychiatric genetics to accomplish her goals as an independently funded investigator. The research outlined in this proposal will provide the necessary training in 1) the design, management, and statistical analysis of clinical databases; 2) survey methodology to capture psychological traits during prospective cohort studies; and 3) GWAS data interpretation and calculation of genetic risk scores. Dr. Nash has built a core mentorship team consisting of Dr. Samantha Meltzer-Brody (co-lead mentor), a physician-scientist and internationally recognized expert in mood disorders; Dr. Patrick Sullivan (co-lead mentor), a world-renowned psychiatric geneticist; Dr. Yun Li (co-mentor), a statistical geneticist who developed core methods for genomics, including imputation and genetic association; and Dr. David Gerber (co-mentor), Chief of Abdominal Transplant at UNC who has published widely in the field including work examining survival benefits and long-term outcomes for transplant patients. Their combined mentorship will allow Dr. Nash to succeed as an independent investigator.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY (See instructions): This project proposes augmented reality eyeglasses that can be worn throughout the day by a person with Parkinson's disease (PD) undergoing neurorehabilitation. The system enables (1) monitoring of and feedback to the patient during clinic visits, with provider-supervised physical therapy (PT) exercises, (2) display of immersive guidance and feedback to the patient during PT exercises at home that reference exercises supervised in the clinic, and (3) monitoring, analysis, and summarization of the patient’s activities of daily life (ADLs) between clinic visits, through cameras integrated into the eyeglasses and inertial sensors in shoes and wristbands. To develop such a system, we will advance the state-of-the-art in four research thrusts - egocentric tracking and reconstruction, egocentric visual perception, personalized reasoning models, and immersive analytics. In the first thrust, we will investigate the use of deep neural network architectures for highly accurate, clinically useful egocentric tracking and 3D reconstruction, enabling us to meaningfully record, playback, and evaluate rehabilitation activities. In the second thrust, we will develop a rich semantic understanding of contextualized activities using multimodal deep learning models that take into account short-term and long-term data from wearable inertial sensors and multiple egocentric cameras. In the third thrust, we will develop novel reasoning systems based on graphical models that leverage techniques such as partially observable Markov decision processes (POMDPs) to track and predict aspects of user performance during rehabilitation activities. In the fourth thrust, we will design an immersive analytics front-end interface that will integrate with our reasoning systems to provide customized user feedback and clinician summaries, enabling improved patient care. Developments across each of these thrusts will be integrated into an eyeglasses-based augmented reality system that will be iteratively evaluated and refined, first with able-bodied participants in a controlled lab setting, next with Parkinson’s patients in a clinical setting, then with able-bodied participants in home settings, and finally with Parkinson’s patients in their home environment.