Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2022-09

The central goal of this R-25 Research Education Program is the creation of a pipeline of innovative and talented problem solvers who can (a) identify the unmet clinical needs of patients suffering from kidney disease, including current and future challenges, opportunities and gaps in kidney care (b) create innovative patient centered solutions (diagnostics, therapeutics, patient centered tools) for kidney disease patients (c) understand the importance of a holistic, multi-disciplinary approach to kidney disease innovation that goes all the way from ideation and investment, to experimental and clinical data, to regulation and reimbursement pathways and (d) leverage the current kidney innovation landscape to develop solutions that actually get to patients so that kidney disease patients will no longer have to look at the parade of therapies for other diseases and wonder “if not me then who and if not now then when…” We plan to achieve these goals by (a) creating a pipeline of multidisciplinary and talented problem solvers (b) developing a summer kidney immersion program (skills development) (c) forming multi-disciplinary problem-solving teams (coached by multi-disciplinary mentors) that do research projects to address unmet clinical needs and (d) nurturing an alumni and retention program for networking and career opportunities respectively. In summary, we will create a holistic, patient centered, multi-disciplinary, team-based approach to problem solving in the kidney technology area, using a robust pipeline of young, innovative, and creative trainees. Most importantly, we plan to invest in mentorship and professional development, that will hopefully keep these trainees in the kidney technology innovation area; and by doing so transform the way that we will care for patients with kidney disease in the future.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY The COVID-19 pandemic has disproportionally impacted rural communities, which are often health professional shortage areas that lack health care infrastructure, including hospitals1-3. When compared to urban populations, individuals living in rural areas are more vaccine hesitant8,9, have a higher prevalence of comorbid health conditions that increase their risk for severe COVID-19 complications12, and are at greater risk from SARS-CoV-2 variants due to lower vaccination rates and fewer vaccination mandates33,40. Thus, interventions to increase vaccine uptake in rural areas are greatly needed. As the most accessible healthcare professional in rural areas4,5 and one of the most trusted sources of medication information16, community pharmacists are well-positioned to address vaccine hesitancy with underserved, rural populations. Because vaccination conversations are sensitive and often politically charged, pharmacists need implementation support, including training and ongoing guidance to deliver evidence-based vaccine hesitancy counseling interventions18. Implementation facilitation, in which trained facilitators coach and troubleshoot problems with professionals as they implement new practices, increases adoption of practices with fidelity23-26. However, implementation facilitation generally, and virtual facilitation (e.g., video coaching) in particular, has not been systematically studied in community pharmacy settings. The need to examine the effectiveness of virtual facilitation as a means to increase vaccine hesitancy counseling and COVID-19 vaccination is of great importance given the large travel distances to rural areas and social distancing. Our goal is to test if virtual facilitation increases rural pharmacists’ ability to implement COVID-19 vaccine hesitancy counseling when compared to a “standard” implementation approach (e.g., training and dissemination of implementation support tools). Using a rural pharmacy practice-based research network (PBRN) that spans 5 southeastern states, we propose two aims. Aim 1 involves a stepped-wedge trial with 30 rural pharmacies to test whether virtual facilitation outperforms the standard approach in increasing: (a) the fidelity with which pharmacists implement the vaccine hesitancy counseling intervention and (b) the number of vaccine hesitant patients who agree to receive the vaccine. Using a project-sponsored data collection system, we will gather data on implementation outcomes, including fidelity and effectiveness. In Aim 2, we will conduct a cost assessment to explore the sustainability of virtual facilitation. We are uniquely positioned to test these competing implementation approaches and leverage rural community pharmacists to engage in vaccine hesitancy counseling in rural communities that have low vaccination rates.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT A Career Development Award in Global Postpartum Health This application is for a Fogarty International Center K01 International Research Scientist Development Award (IRSDA) for Dr. Bridget Spelke, MD, an obstetrician-gynecologist and research fellow in Global Women's Health at the University of North Carolina (UNC). Dr. Spelke is in the formative stages of her research career and aims to establish herself as an independent investigator and future leader in the area of global postpartum health, a high-risk period that has been neglected for too long. She has lived full-time in Lusaka, Zambia since August 2019 where she has established a solid career foundation for clinical research in obstetric outcomes for women in resource-constrained settings. Dr. Spelke's proposed K01 research will leverage a recent expansion of the Zambian Preterm Birth Prevention Study (ZAPPS), a long-standing pregnancy cohort that is following women and their infants through 12 months postpartum. Dr. Spelke will study a novel strategy to identify women at risk for severe maternal morbidity and mortality in the first week after delivery: continuous remote physiologic monitoring with easy-to-use, multiparameter, wearable sensors. To inform the design of a future hybrid effectiveness-implementation trial, Dr. Spelke proposes a mentored research program that will estimate the frequency of severe maternal morbidity in the ZAPPS cohort (Aim 1); implement a pilot randomized feasibility trial of postpartum physiologic monitoring (Aim 2); and evaluate barriers and facilitators to adoption of a digital health intervention in a resource-constrained setting through implementation research frameworks (Aim 3). These three research aims will provide preliminary data toward the design of a future trial. They are interrelated and independent of each other; study results will advance research on digital and postpartum health in resource- constrained settings regardless of outcomes. Each of these studies aligns with individualized career development goals in clinical trials design and analysis, implementation research for digital health technologies, and collaborative global health research partnerships. Dr. Spelke has assembled a highly accomplished and eminently qualified mentorship team comprising: her primary Zambia mentor, Prof. Bellington Vwalika, an expert in clinical trials implementation in sub-Saharan Africa; her primary US mentor, Prof. Elizabeth Stringer, an expert in maternal-fetal medicine and maternal health epidemiology; and her core mentor, Prof. Ben Chi, an expert in implementation science in sub-Saharan Africa. All three mentors have decades of experience in collaborative global health partnerships and extensive experience mentoring early career researchers. The IRSDA will advance Dr. Spelke's skills in each of these fields, supporting both didactic and practice-based learning opportunities, as she establishes her independent area of investigation and progresses towards research independence in global postpartum health.

NIH Research Projects · FY 2025 · 2022-09

Abstract Accurate replication of the genome during cell proliferation is necessary for normal animal development and homeostasis. Disruption of the regulation or fidelity of replication contributes to many human pathologies, particularly cancer. Thus, a complete understanding of the mechanisms governing genome replication is paramount to human health. Replication of large genomes like that found in human cells requires the initiation of bi-directional DNA synthesis at thousands of individual locations on each chromosome. It also requires cell cycle-regulated synthesis of large amounts of histone proteins to package newly replicated DNA into chromatin. This project will focus on how chromatin assembly and organization influences genome replication during animal development. The basic building block of chromatin is the nucleosome, an octamer of histone proteins encompassed by ~147 base pairs of DNA. Most histone proteins within chromatin are synthesized each S phase of the cell cycle from replication-dependent histone genes, which encode the only eukaryotic mRNAs that end in a stem loop rather than a poly A tail. The genes encoding all five RD-histone proteins are clustered in metazoan genomes, and transcription and pre-mRNA processing factors required for histone mRNA biosynthesis are organized into a nuclear body (the Histone Locus Body or HLB) that assembles at these gene clusters. We will determine the requirements for the coordinate synthesis of the RD-histone mRNAs using both biochemical and genetic approaches in Drosophila, with a particular focus on the role that the HLB plays in histone transcription and pre-mRNA processing. Each core histone protein has an N-terminal tail that protrudes from the nucleosome core and is subject to a variety of chemical modifications (e.g. methylation, acetylation, and phosphorylation) that modulate chromatin organization and thus influence all aspects of genome function, including DNA replication. We have developed a method in Drosophila for engineering any desired histone tail mutation, providing us a means of preventing modification of specific histone residues and thus of manipulating chromatin organization in a way not available in any other animal. This genetic approach will be combined with cell biological and next generation DNA sequencing methods to determine how chromatin organization modulates DNA replication throughout the entire genome, thereby addressing a major question in the field. Our Drosophila experimental paradigm permits the in vivo interrogation of these fundamental processes in gene expression and DNA replication in ways that are unavailable in other experimental systems.

NIH Research Projects · FY 2025 · 2022-09

Unhealthy alcohol use is common among people using pre-exposure prophylaxis (PrEP). Unhealthy alcohol use is a spectrum from risky/hazardous drinking to alcohol use disorder. For people on PrEP, unhealthy alcohol use may reduce PrEP persistence and adherence, which are critical for HIV prevention. Reducing unhealthy alcohol use may improve PrEP outcomes. The brief alcohol intervention (BAI), which draws on Motivational Interviewing and Cognitive Behavioral Therapy, increases viral suppression among people living with HIV on antiretroviral therapy. This effect is mediated through increased adherence. The BAI may have a similar effect on PrEP outcomes among people with unhealthy alcohol use. We propose an effectiveness-implementation type 1 hybrid randomized controlled trial (RCT) to compare the BAI to standard of care among people with unhealthy alcohol use initiating or re-initiating PrEP in Vietnam. We hypothesize that the BAI will improve a) PrEP persistence, b) PrEP adherence, and c) risk-aligned PrEP use (PrEP use consistent with current risk, including event-driven PrEP). Our specific aims are to: 1) Assess effectiveness of the BAI for increasing PrEP persistence and adherence among people in Vietnam; 2) Assess the impact of the BAI on alcohol use among people using PrEP; 3) Estimate the cost-effectiveness, feasibility, and acceptability of scaling up the BAI in PrEP clinics throughout Vietnam. The trial and intervention will be conducted at the Hanoi Medical University PrEP clinic in Vietnam. Evaluation visits will be conducted at UNC Project-Vietnam. Eligible participants will include people (n=564) 16 years of age or older presenting to the PrEP clinic for PrEP initiation or re-initiation after at least 3 months from a missed PrEP appointment, based on local PrEP guidelines. The participants must also screen positive for unhealthy alcohol use with an AUDIT-C score ≥ 4 or have a CRAFFT score ≥ 2 (for those ≤21 years old). After baseline evaluations, participants will be randomized to either standard of care or the BAI, which comprises two in-person and two telephone visits. Each participant will be followed quarterly with interviews, including use of timeline followback to elicit alcohol use, and tests for HIV infection. Alcohol use and PrEP adherence will also be assessed with biomarkers. The primary effectiveness outcomes are PrEP persistence and heavy drinking days. Secondary outcomes include PrEP adherence, assessed by self-report, pharmacy records, and validated bioassays; risk-aligned PrEP use; and alcohol outcomes (number of drinking days; and drinks per drinking day). The primary implementation outcomes are the incremental cost-effectiveness ratio, expressed as the incremental cost per quality-adjusted life-year gained, and feasibility and acceptability. If successful, this study will be set to ensure rapid scale up of an intervention to improve lives by reducing unhealthy alcohol use and increasing PrEP use for HIV prevention.

NIH Research Projects · FY 2025 · 2022-09

This is an application for a cooperative agreement (U54), “Partnerships to Enhance Alcohol Research across NCCU and UNC (PEAR-NC),” between faculties of North Carolina Central University (NCCU), and the Bowles Center for Alcohol Studies (BCAS) at the University of North Carolina (UNC) School of Medicine. This proposal represents a true collaborative effort between the NCCU and UNC faculty with both groups contributing significant effort and being essential to the success of this Alcohol and chronic disease research program. Within this proposal, an NCCU Administrative Core and three Research Components will integrate with the UNC-BCAS Administrative Research Components, as well as other activities at the Bowles Center for Alcohol Studies. The UNC NIAAA Alcohol Research Center (ARC) and BCAS have many faculty studying mechanisms of alcohol pathology across the spectrum of behavioral, tissue, and cellular pathologies that occur with alcohol exposure. This proposal will focus on cellular pathologies that allow collaborative research partnerships on common areas of interest between UNC and NCCU faculty. These partnerships open UNC research laboratories, core facilities, and medical libraries to NCCU faculty, improving opportunities for developing productive research laboratories at NCCU. In addition, due to recent NCCU NIAAA R awards, large numbers of NCCU students will continue to become involved in research on mechanisms of alcohol pathology, providing education on alcohol-related health outcomes and comorbidities . The objective of this U54 partnership is to investigate molecular mechanisms of alcohol-induced cellular pathology. This proposal includes three NCCU research projects, involving six NCCU faculty, that will investigate the molecular mechanisms of chronic diseases including fetal alcohol pathology, alcohol-induced mammary stem cell toxicity in cancer, and chronic neuroimmune responses in adolescent and adult brain. A second objective is to provide research experiences and scholarly education on alcohol pathology for NCCU students. The educational efforts in this proposal will involve NCCU undergraduate and graduate students through the BCAS seminar series, through training in UNC laboratories and research development and mentor meetings, attendance and presentation of data at the annual RSA meeting, an alcohol seminar series hosted by NCCU, new course curricula on alcohol pathology developed with a focus on alcohol-related health outcomes, as well as specific undergraduate student summer internships for NCCU students to study and learn about alcohol pathology within laboratories in both institutions. Ultimately, this proposal will conduct, promote, support, and mentor research into mechanisms of alcohol pathology, creating an active and successful alcohol research program within NCCU that synergizes with UNC to advance education and discoveries.

NIH Research Projects · FY 2025 · 2022-09

Abstract Enzymes are powerful biocatalysts that may be repurposed for efficient and sustainable synthesis of industrial and pharmaceutical compounds. Identifying novel enzyme-catalyzed reactions and understanding their catalytic mechanisms is essential for expanding the biocatalytic toolbox. We have recently discovered a unique copper-containing antibiotic, fluopsin C, which is produced by Pseudomonas aeruginosa, a leading cause of multidrug-resistant nosocomial infections. By studying how P. aeruginosa makes fluopsin C, we identified an unusual biosynthetic pathway that involves novel enzyme chemistry. For instance, two iron-dependent enzymes of a new heme- oxygenase-like enzyme family catalyze a carbon excision and an oxidative decarboxylation and N-hydroxylation respectively. Two lyases of the adenylosuccinate lyase enzyme family exhibit novel activities of making and breaking carbon-sulfur bonds respectively. The chemistries we identified have expanded the catalytic capabilities of these enzyme families. We hypothesize that the fluopsin C biosynthetic enzymes use novel catalytic mechanisms and can be repurposed or engineered to generate new molecules. To test this hypothesis, we will use a combination of enzyme kinetics, structural biology, spectroscopy, and synthetic chemistry tools, with a focus on three enzymes in fluopsin C biosynthesis. Studies of these distinct enzymes will advance the fundamental understanding of how their respective enzyme family catalyzes diverse chemistry. Defining the mechanisms and substrate scope of the enzymes will allow them to be further explored for biocatalytic use.

NIH Research Projects · FY 2025 · 2022-09

Project Abstract Despite the high global prevalence of knee osteoarthritis (OA), treatment options have been limited to symptom management and total joint replacement in large part because the mechanisms driving knee OA remain poorly understood. Genome-wide association studies (GWAS) suggest that non-coding genetic variation is a major contributor to knee OA disease-risk; however, the LD structure of human genomes, the long-range nature of transcriptional regulation, and the lack of genome-editable and biologically accurate systems in which to study OA have created a bottleneck that hinders our ability to translate GWAS findings into new treatments. The overall objective of this proposal is to identify putative causal knee OA risk variants, map them to their target genes, and quantify their phenotypic impact in chondrocytes. We will identify regulatory regions activated in response to cartilage degradation (Aim 1), map regulatory loci and GWAS variants to their target genes (Aim 2), and quantify the phenotypic impacts of knee OA-associated variants using an ex vivo model of the chondrocyte OA phenotype (Aim 3). This work will break down existing barriers in knee OA genetics, improve our mechanistic understanding of knee OA, and provide new risk genes for further study and therapeutic development.

NIH Research Projects · FY 2025 · 2022-09

Human parainfluenza virus (PIV) and metapneumovirus (MPV) are common causes of lower respiratory tract infections in infants and young children, and major causes of respiratory illness in immune compromised adults and the elderly. Unfortunately, there are no therapy or vaccine for either viruses, and only supportive medical care is available. Interestingly, PIV and MPV share many pathological and clinical manifestations as Respiratory Syncytial Virus (RSV). Indeed, neither PIV nor MPV are known to cause viremia in the blood of infected patients, indicating both infections are strictly localized in the airways similar to RSV that shed progeny viruses exclusively from the apical surface of infected cells. Such unique pathophysiology implies progeny viruses must traverse airway mucus (AM) before spreading to neighboring cells. This in turn motivated us to develop a pathogen-specific antiviral that could physically limit the spread of the infections within the airways. We recently discovered a novel Ab effector function in mucus – trapping individual pathogens in mucus based on carefully-tuned affinity between IgG-Fc and mucins – and developed a platform for enhancing mAb function at mucosal surfaces. We hypothesize that “muco-trapping” mAb delivered to the airways can directly intervene with the viral life cycle by intercepting and trapping shed progeny viruses in AM, rapidly eliminate trapped viruses from the airways by natural mucociliary clearance, and enable effective therapy in vivo. In support of this strategy, we engineered mAbs that potently trap RSV in AM, and showed that nebulized delivery of “muco-trapping” mAb to RSV-infected neonatal lambs beginning on Day 3 post infection effectively reduced the infectious viral load in lung tissues to non-detectible levels within 3 days, with viral RNA in bronchial tissues reduced by 11-fold compared to vehicle control. These results motivated us to explore whether a similar approach may be effective in treating PIV and MPV infections. In support of this application, we have engineered antibodies with picomolar affinity to diverse strains of both viruses, and demonstrated that we could effectively trap viruses in human AM and limit spread of infection in vitro in well-differentiated human airway epithelium (WD-HAE) grown at the air-liquid interface. In this proposal, we will continue our work with affinity maturation using mammalian and yeast display to produce high affinity mAb that broadly bind and neutralize diverse strains of PIV and MPV (Aim 1). We will validate whether these mAbs can trap PIV and MPV in fresh undiluted human AM, and whether they can inhibit the spread of pre-established PIV and MPV infections in WD-HAE cultures (Aim 2). We will then advance the lead mAb for both viruses for evaluation in a hamster nasal infection model (Aim 3). By enabling enhanced mAb function in mucus secretions, we expect we will help pave the way for improved, molecularly-targeted therapies and prophylaxis against a broad spectrum of pathogens across all major mucosal surfaces, providing a powerful option addressing the current gap in pharmacological interventions for respiratory infections.

NIH Research Projects · FY 2025 · 2022-09

SToP CANCER SPORE ABSTRACT Pancreatic cancer remains a lethal disease with limited therapeutic options. Options have increased over the last 5 years, with large genomic analyses and preclinical efforts. However, despite advances, pancreatic cancer remains a lethal disease with a five-year survival rate of 10%, and deaths from pancreatic cancer are expected to surpass deaths from breast, prostate and colorectal cancer by 2030, to become the second leading cause of cancer deaths. The incidence is rapidly increasing, with, a 23% increase since 2010 and 57% increase since 2006. The etiology and reason for the recent rise remain poorly understood with a complex interplay of somatic genetic, genomic, epigenetic and environmental factors in the context of an aging population. Unlike many solid tumors, mutations alone have not been sufficient to yield curative targeted therapies, clinically useful biomarkers or consensus subtypes. Challenges that remain include: · Low tumor cellularity hampers both genetic and genomic studies, including the inability to identify biomarkers/subtypes to tailor therapies · Chemotherapy and targeted therapy resistant tumor cell populations · Desmoplastic stroma that may be both tumor promoting and therapy inhibiting · Immunosuppressive environment due to suppressive myeloid cells and a paucity of T effector cells · Precision oncology approaches are still limited · Clinical trials are limited to only 1-2 therapies at a time We have assembled three projects that directly address these issues; each has an embedded early phase clinical trial that will yield patient samples with which to test our SPORE’s hypotheses. Robust Development Research and Career Enhancement Programs are included based on the highly successful models at UNC Lineberger. These will be backed by a substantial institutional commitment. A highly accomplished multidisciplinary team of investigators with collaborations across several institutions have been brought together that includes those who have made innovative and high impact contributions, delivered clinical care and performed clinical and translational research germane to pancreatic cancer. Recognizing the need for pancreatic cancer rapid translation to the clinic, our Tissue Procurement, Pathology, and Genomics Core and Integrative Quantitative Sciences Core work seamlessly with the projects to process and analyze data. Our SToP Cancer SPORE goal is to establish a new paradigm for clinical trial design that is not limited to a single therapy or biomarker.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY More than 25% of people with HIV (PWH) globally report unhealthy alcohol use, defined as a spectrum of use from risky/hazardous (drinking more than the recommended daily, weekly or per-occasion amounts resulting in increased risk for health consequences) to alcohol use disorder. PWH with unhealthy alcohol use may not adhere to their antiretroviral therapy (ART), leading to unsuppressed virus and increasing the potential for HIV transmission to sexual and injection partners. Interventions that address alcohol use among PWH are needed to prevent HIV transmission. We previously showed that the brief alcohol intervention (BAI) is effective at reducing alcohol use and increasing viral suppression in Vietnam. The BAI is ready for scale-up but one barrier to scale-up is attitudes towards alcohol interventions among clinical staff, especially in regions with normative unhealthy alcohol use. Addressing clinical staff attitudes may be essential for scale-up. We propose a hybrid type 3, cluster randomized implementation trial to examine effective strategies to scale up the BAI in ART clinics in Vietnam. One arm will receive only facilitation for BAI implementation. Facilitation is a flexible strategy that helps clinics to address common barriers, such as counselor skills, competing priorities, and resource deficits. In the other arm, in addition to facilitation, clinic staff, irrespective of their own alcohol use, will be offered the BAI themselves as experiential learning to address their own alcohol-related attitudes and behaviors. We hypothesize that EBAI, added to facilitation, will increase BAI fidelity, acceptability, and penetration at the clinic level, and improve viral suppression among PWH with unhealthy alcohol use. Our specific aims are to: 1) Compare BAI implementation using facilitation (FAC) only to experiential BAI plus facilitation (EBAI+FAC) in ART clinics in Vietnam; 2) Explore the mechanisms of successful BAI scale up in both the FAC and EBAI+FAC arms; and 3) Measure the impact of EBAI on clinic staff. ART clinics (n=30) across Vietnam will be randomized to receive FAC or EBAI+FAC. PWH in the clinics will be screened for unhealthy alcohol use with the AUDIT-C; if positive, they will be offered the BAI. The primary implementation outcomes are clinic-level BAI fidelity (primary), with secondary outcomes of acceptability, penetration, and cost. The effectiveness outcomes are viral suppression (primary) and alcohol use (secondary), measured among a cohort of PWH recruited in each clinic. Outcomes will be measured at 3 months (implementation outcomes only) and 12 months (all outcomes). In parallel with the trial, we will use mixed methods to examine the organizational and clinic staff characteristics that underly successful BAI scale-up. Given the importance of the clinic staff in the BAI implementation, we will also explore the impact of the BAI on their own alcohol-related attitudes and use and considering whether they received the BAI themselves or not. This trial will present critical information for worldwide HIV treatment as prevention efforts, providing strategies for effective scale-up of the BAI among PWH with unhealthy alcohol use.

NIH Research Projects · FY 2025 · 2022-09

Risk for major depressive disorder (MDD) onset dramatically increases during adolescence, particularly for girls. There is a vital need to elucidate the underlying mechanisms of MDD risk to improve early risk detection and targeted preventions. The stress generation model posits interpersonal stress generation (I-StressGen) as a mechanism of risk and proposes that youth with preexisting MDD vulnerabilities actively contribute to the occurrence of interpersonal stressful life events in their lives (i.e., I-StressGen), which in turn exacerbates their risk for MDD. Although the impact of I-StressGen on prospective MDD risk in adolescents is well-established, the precise mechanisms contributing to I-StressGen remain unclear. Self-report research indicates increased affective reactivity during negative social interactions as a promising mechanism contributing to I-StressGen, though this has not been directly tested. The underlying neural mechanisms of I-StressGen also remain unexamined, despite work suggesting that greater social stress is associated with aberrant activation in corticolimbic regions during tasks probing response to negative social interactions. Therefore, the overarching goal of the proposed study is to obtain a fine-grained, mechanistic understanding of the role of brain and behavioral affective reactivity during negative social interactions as it relates to I-StressGen and subsequent MDD risk in female adolescents. The proposed study will enroll a risk-enhanced sample of 90 female adolescents (ages 13-15) and will utilize neuroimaging (fMRI) and ecological momentary assessment (EMA) to examine brain and behavior indices of socio-affective reactivity as predictors of I-StressGen and depression symptoms across a multi-wave one year follow-up. In doing so, the current study will test whether affective reactivity during negative social interactions at the neural (Aim 1) and real-world behavioral (Aim 2) level prospectively predicts I- StressGen, and whether I-StressGen mediates the relation between this socio-affective reactivity and prospective increases in female adolescents’ depression symptoms (Aim 3). The proposed study and parallel training plan will allow the Candidate to build upon her current expertise in adolescent stress and MDD risk and develop new skills in four key areas: (1) fMRI, (2) EMA, (3) advanced statistical multilevel modeling, and (4) professional development. The rich academic environment at the University of Illinois at Chicago coupled with the collective expertise of the Candidate’s mentorship team in fMRI methodology, developmental affective neuroscience, EMA, and advanced statistical techniques will facilitate successful implementation of the proposed study and training plan. The acquisition of these skills will be integral to launching the Candidate’s independent career as a translational scientist focused on delineating brain-behavioral mechanisms of adolescent MDD risk within the context of real-world interpersonal stress. Results from this study will inform the development of future grants focused on characterizing socio-affective brain and behavior mechanisms and aberrant developmental trajectories contributing to the emergence of maladaptive stress processes and MDD onset in adolescence.

NIH Research Projects · FY 2025 · 2022-09

Abstract Congenital heart disease (CHD) remains the most common congenital malformation. Therefore, attaining a mechanistic understanding of cardiomyocyte formation is crucial for improving outcomes to structural heart disease. Post-translational modifications of histones act to regulate cardiac chromatin structure and hence, the temporal and spatial program of gene regulation during cardiac development. We have found that SMYD1, a cardiomyocyte essential histone methyltransferase interacts with the chromatin remodeling MLL4 class of Complex of Proteins ASsociated with Set1 (COMPASS) complex. SMYD1 has been shown to be essential for cardiac development and as we show here, causes CHD. Like SMYD1, two of the core components of the MLL4-COMPASS complex, KMT2D/MLL4 and KDM6a are essential for cardiac development and cause CHD. The goal of the current application is to test the central hypothesis that SMYD1 acts within the MLL4-COMPASS complex to clear histones at cardiac enhancers prior to gene activation. This will be achieved by: 1) 2) Determining the function of SMYD1 in the assembly and function of the cardiac MLL4- COMPASS complex and, 2) Establishing the requirement for KDM6a in MLL4- COMPASS activity. Collectively, these studies will provide a detailed mechanistic understanding of the tissue specific role for SMYD1 and MLL4-COMPASS complex in cardiac development and heart disease.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY This proposal will address the critical need for new and modifiable targets to enhance mobility and restore independence to those in our rapidly aging population. Due to reduced ankle push-off power, older adults walk slower and with higher metabolic energy cost than younger adults. As our central premise, we contend that hallmark age-associated deficits in push-off intensity during walking have been far too often mistakenly attributed solely to the plantarflexor muscles, and instead originate interdependently with those in the active, passive, and structural regulation of foot mechanical power. This premise paves the way for translational opportunities to augment foot structure and function to enhance independence and quality of life. This study combines the research agendas of two highly productive investigators and leverages the research infrastructure of two peer institutions. Aim 1 will be the first to study mechanical power interactions between the human foot and ankle in governing reduced push-off intensity and walking economy in older adults across a wide variety of everyday walking tasks. By combining metabolic measurements with state-of-the-art biomechanical and bioenergetic modeling, we will test the hypothesis that older adults exhibit higher mechanical energy losses via foot structures than young adults – aging effects that: (i) are larger for walking tasks that increase foot demand, (ii) misappropriate ankle moment and power during push-off, and thereby (iii) correlate with shorter 6 min walk distance and increased metabolic energy cost compared to young adults. Aim 2 will provide mechanistic insight into aging effects on the active, passive, and structural regulation of foot-ankle mechanical power interactions during walking. Using a series of controlled loading paradigms on a dynamometer combined with advanced in vivo ultrasound imaging and novel electromyographic biofeedback, we will test the hypotheses that older adults exhibit: (i) reduced foot and plantarflexor muscle strength and (ii) lower structural stiffness of and (iii) reduced structural connectivity between series elastic tissues spanning the foot and ankle – changes that require elevated plantar intrinsic muscle activation to maintain requisite foot stiffness and associate with reduced ankle moment and power output during push-off in walking. Finally, as a translational benchmark, Aim 3 will show that shoe- stiffness modifications that act in parallel with the plantar aponeurosis and intrinsic muscles can mitigate age- associated deficits in push-off function during walking. Supported by promising pilot data, we will test the hypotheses that older adults walking with increased shoe insole stiffness will exhibit: (i) smaller mechanical energy losses at the foot, (ii) more favorable plantarflexor muscle contractile dynamics, (iii) greater peak ankle moment and power output, and thus (iv) longer 6 min walk distance and reduced whole-body metabolic energy cost. Ultimately, this work will establish a paradigm shift in our biomechanical understanding and clinical management of age-related mobility impairment toward feasible and cost-effective devices to modify foot-ankle function – an outcome with significant potential to enhance independence and quality of life for millions.

NIH Research Projects · FY 2025 · 2022-09

Acute and chronic kidney diseases (CKDs) are often interrelated, and each can predispose to the other. Both are common and costly yet biomarkers and treatments are incomplete. The Kidney Precision Medicine Project (KPMP) addresses this problem via acquisition of research protocol kidney biopsies for pathologic and biomolecular interrogation. This application responds to RFA-DK-20-026, proposing addition of the Universities of North Carolina (UNC) and Arizona (UA) as highly resourced recruitment sites enriched with rural, Black, LatinX and indigenous populations and with renowned centers in health equity and environmental health research. Inclusion of geographically diverse populations is critical to KPMP, as socioeconomic determinants of health represented by neighborhood context and environmental pollutants explain much of the geographic variability of CKD prevalence. ▪ AIM 1: To adhere to the highest safety and ethical standards, while successfully recruiting and retaining KPMP participants from geographically, racially, and ethnically diverse populations with AKI and/or CKDs. We have included experts in bioethics and health equity to inform and guide our approaches and overcome barriers to recruitment and retention. To provide the highest safety for our KPMP participants, we have partnered with interventional radiology to perform all research kidney biopsies. ▪ AIM 2: To fully engage and collaborate with investigators of the KPMP to maximize its efficiency, productivity, and discovery with the goal of reducing the human and economic toll of CKDs and AKI. We will adhere to existing protocols of KPMP, actively participate in committees and writing groups, and provide creative strategies to troubleshoot barriers and optimize workflow within KPMP. ▪ AIM 3: To enhance the breadth of KPMP scientific productivity through inclusion of fully diverse populations and acquisition of detailed environmental exposure data that are currently not captured. There is geographic heterogeneity in the prevalence and severity of CKDs. Using new tools, our site will provide new information about environmental exposures for CKDs and AKI that will broaden the interpretation of KPMP findings. In summary, our application proposes to further strengthen KPMP through demographic and geographic diversification using tools to assess neighborhood context and identify environmental exposures to expand research opportunities in these fields critical to the pathogenesis of AKI and CKDs.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Genomic sequencing offers an unprecedented opportunity to identify clinically relevant genetic variants, yet there are many challenges to overcome before this technology can be applied routinely in the healthy population to identify individuals with actionable disorders in time to prevent or ameliorate symptoms. This project explores an innovative age-based genomic screening (ABGS) paradigm that aims to provide targeted and highly actionable genetic information to parents via their child’s routine wellness visits, thereby avoiding some of the more difficult aspects of genome-scale sequencing while retaining most of the benefits. We will utilize an established metric that evaluates parameters relevant to “clinical actionability” and defines the age of onset and/or the age of intervention, to identify conditions that would be applicable for screening. In collaboration with a diverse Expert Deliberative Group, we will define a consensus framework for carrying out the ABGS program using targeted sequencing panels at specified time-points during infancy and childhood. We will engage community stakeholders to raise awareness of the ABGS program and obtain critical feedback to inform the development of accessible study materials. We will apply the Genetic Medicine Implementation Research framework and utilize rigorous methods and measures to identify potential barriers and facilitators and develop strategies to address them. Finally, we will conduct a pilot project in a small number of primary care pediatrics clinics assessing preliminary outcomes, including perspectives of parents and providers, and the feasibility, acceptability, and utility of ABGS. The expected result of this proposal is a validated, stakeholder-informed, and practical ABGS program that includes hundreds of conditions that are actionable throughout the lifespan, setting the stage for a future longitudinal study in a larger number of practices that can assess clinical and health economic outcomes. Throughout this work, we will employ a community-based participatory research approach to seek out perspectives from diverse stakeholders and emphasize the importance of studying ABGS in a wide range of settings. The long-term goal of this research program is to create a broadly applicable genomic screening program that extends well beyond newborn screening and can be incorporated into routine well child care. We envision that this program will also prepare individuals to eventually make informed decisions about the potential benefits and risks of screening for adult-onset conditions during adulthood, thus creating a connection between genomic screening efforts in all age groups.

NIH Research Projects · FY 2025 · 2022-09

Tobacco use remains the leading cause of preventable chronic diseases and death in the United States. Almost 50% of youth tobacco users now concurrently use two or more tobacco products—defined as multiple tobacco product (MTP) use. Youth MTP use is concerning because it is associated with a two-fold increase in nicotine dependence symptoms, decreased likelihood of successful quitting, and increased other substance use. Many communication campaign ads exist to warn youth of the harms of specific tobacco products, like e-cigarettes or cigarettes. However, almost all ads focus on one tobacco product at a time, which could lead MTP users to consume the tobacco product that is not the target of the ads. The overarching goal of the proposed K01 project is to develop and evaluate messages about the harms of MTP use. The primary hypothesis is that MTP messages will increase intentions to quit using all tobacco products more than existing messages that focus only on single tobacco products and control messages. In Aim 1, I will identify effective themes for youth-oriented messages that communicate the harms of MTP use. In Aim 2, I will develop a set of high impact messages that discourages MTP use among youth and determine whether other message elements (e.g., testimonials, quitting approach, self-efficacy cue) increase tobacco quit intentions using a discrete choice experiment with youth MTP users. Finally, in Aim 3, I will conduct a three-arm pilot randomized controlled trial (RCT) with youth MTP users to see if the MTP messages delivered as text messages over a two-week period increase tobacco quit intentions more than existing messages that focus only on single tobacco products and control messages. This research would make significant and timely contributions to the development of tobacco control strategies in the United States and is responsive to FDA’s interest in communications research. My long-term career goal is to conduct independent research that focuses on preventing and reducing youth tobacco use through health communication interventions. This career development award will help me reach this goal by providing new training in a) health communication theory and message development, b) experimental design for health communication research, including design of discrete choice experiments, and c) eHealth interventions for youth. My training plan includes apprenticeships with my mentorship team, formal coursework, hands-on research activities, and workshops. This award will culminate in an R01 application for a full-scale RCT testing the effectiveness of MTP messages on quit attempts and other behavioral outcomes among youth MTP users.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT People with serious mental illnesses (SMI) are disproportionately represented throughout the criminal justice system, the vast majority of whom are on probation and parole. Implementing evidence-based practices to address the needs of justice-involved people with SMI requires an understanding of the multilevel factors that impact intervention implementation in both the mental health and criminal justice systems and the expertise to address these factors. Dr. Tonya Van Deinse’s proposed K01 training plan and research study, under the expert guidance of an NIH-funded mentoring team, will provide her with intensive substantive and methodological training and research experience to become a leading expert in implementing complex, cross- agency interventions to address the needs of justice-involved people with SMI. Specialty mental health probation (SMHP) is an evidence-informed intervention that aims to engage people with SMI in treatment and divert them from the criminal justice system. Although high-fidelity SMHP can increase mental health and substance use service engagement, improve mental health symptoms, and reduce rates of rearrest, jail days and probation violations, effective implementation of SMHP is context-dependent and relies on inter- organizational relationships between SMHP officers and mental health service providers. Specifically, SMHP requires officers to increase treatment linkage and resource coordination and serve as boundary spanners who bridge the interagency coordination and collaboration gap between probation officers and mental health service providers. Given the importance of inter-organizational relationships in implementing SMHP, it is necessary to assess the multilevel factors that impact its cross-agency implementation context and to develop implementation strategies to enhance inter-organizational relationships. This proposal adapts an existing implementation approach to enhance probation officers’ capacity for coordination, collaboration, and linking probationers with SMI to treatment. This adapted implementation approach, to be called Networking to Engage in Treatment: Working within Officer-Resource Collaboratives (NETWORC), will be developed and tested via the following aims: (1) identify multi-level factors that inhibit SMHP officers’ capacity to link individuals with SMI to treatment providers; (2) conduct a process evaluation of the NETWORC implementation approach; and (3) evaluate the efficacy, acceptability, appropriateness, and feasibility of the collaborative implementation toolkit. Findings from this study will provide preliminary data about the feasibility and efficacy of the adapted implementation strategy and will be used to develop Dr. Van Deinse’s subsequent R01, a pragmatic cluster randomized trial using multilevel analyses to examine the impact of the implementation strategy on probationer, officer, and inter-organizational levels.

NIH Research Projects · FY 2025 · 2022-09

The proliferation of high-throughput technologies has led to previously unimaginable growth in biomedical research data sets and knowledgebases. Nearly all these data and knowledge sources address specialized areas of biomedical research, leading to natural diversity but also growing disintegration between individual knowledgebases. This trend generates downstream inefficiencies when applying analytics to enable actionable knowledge discovery from databases. Growing efforts, both in academia and industry, are focused on the development of methods and tools to enable semantic integration and concurrent exploration of disparate biomedical knowledge sources. Recent innovations include the development of biomedical `knowledge graphs' (KGs) that support knowledge discovery through the application of querying and reasoning algorithms and tools. Our team has contributed to these efforts by developing a KG-based question-answering system termed Reasoning Over Biomedical Objects linked in Knowledge-Oriented Pathways (ROBOKOP). Herein, we propose synergistic research and development efforts that aim to significantly advance the ROBOKOP graph knowledgebase capabilities to contribute to high-impact applications across diverse biomedical research domains. Our overarching goal is to equip users with a unique and comprehensive knowledgebase system that supports the rapid generation of mechanistic hypotheses that can explain, validate, or predict biomedical phenomena. We will achieve our objectives by executing studies planned under the following Specific Aims: Aim 1. Enrich and Enhance the ROBOKOP graph knowledgebase. We will enhance the data and infrastructure of the ROBOKOP KB. Aim 2. Provide tools to explore the ROBOKOP graph knowledgebase. We will enhance the ROBOKOP KG by developing and employing novel reasoning tools for KG mining and edge inference. Aim 3. Prove utility and promote use of the ROBOKOP graph knowledgebase through impactful use cases. We will conduct several collaborative proof-of-concept research applications in diverse biomedical domains and diseases. We will actively promote community engagement, user acceptance, and broader impact of ROBOKOP. We expect that our diverse, cutting-edge approach to research, development, and community engagement, coupled with our high-impact biomedical applications, will lead to the formation of a core group of regular users, promote long-term sustainability, and generate impactful new scientific knowledge and mechanistic hypotheses for subsequent testing.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT The Director-General of the World Health Organization has called for invasive cervical cancer (ICC) elimination by 2030. Women living with HIV (WLWH) have a ~10-fold higher risk of ICC. There is an urgent need for high quality evidence on optimal strategies for ICC prevention directly applicable to WLWH who face a disparate burden of cervical precancer and ICC. We will establish a CASCADE Network UG1 Research Base (RB) at the University of North Carolina-Chapel Hill (UNC) to prevent ICC among WLWH. The overarching goal of this UNC RB proposal is to develop, test, and implement effective, scalable, and cost-effective screening and treatment strategies to reduce ICC among WLWH. We will develop, implement, and analyze innovative hybrid effectiveness-implementation trials in the CASCADE Network focus areas of “Improving the Management of Screen Positives” and “Optimizing Precancer Treatment”. The UNC RB will be composed of clinicians, epidemiologists, biostatisticians, and training experts with decades of research experience on ICC prevention and HIV in both LMIC and the US. Our multi-disciplinary team has expertise in implementation science, screening with automated visual evaluation (AVE), pathology, and cost-effectiveness analyses. In Aim 1, we will Develop CASCADE Network trials of evidence-based interventions to optimize the screening for and management of cervical precancer among WLWH by 1) providing scientific, clinical and statistical expertise to develop innovative concepts and protocols for novel, yet pragmatic, hybrid effectiveness- implementation trials, and 2) creating organized processes to efficiently develop concepts and protocols by leveraging diverse input from the UNC RB Scientific, Statistical, Clinical Sites Advisory, Patient Advocacy Advisory, and Scientific Advisory Committees. As an illustrative example of a potential clinical trial for “Improving Management of Screen-Positives”, we propose Protocol 1, a stepped wedge cluster randomized trial to Evaluate AVE triage versus HPV-and-treat among HPV-positive WLWH for CIN2+ prevention. For “Optimizing Precancer Treatment”, we propose Protocol 2, a Hybrid Type 1 trial to evaluate the effectiveness of adjuvant, self- administered intravaginal topical therapy following primary 2/3 treatment to prevent CIN2+ recurrence among WLWH. Outcomes for both trials will include clinical effectiveness for precancer, implementation outcomes, and cost effectiveness analyses. In Aim 2, we will ensure successful Implementation and Analysis of CASCADE Network trials by providing scientific, clinical and statistical expertise, operational support, and regulatory oversight. In Aim 3, we will provide Rigorous Foundational Clinical and Research Training for UG1 Clinical Sites and Early-Stage Investigators to design and implement clinical effectiveness research. If successful, CASCADE Trial findings will advance clinical screening and treatment guidelines for WLWH in LMIC and United States.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract Almost one million newborns die each year from failure to breathe at birth. Nearly all of these deaths occur in low and lower-middle income countries (LMICs). These deaths result when life-saving bag mask ventilation (BMV) is delayed or interrupted. Simulation-based training is commonly used to improve BMV, but gaps in performance remain. There is strong scientific premise for improving BMV with feedback strategies. In randomized simulation trials, feedback during BMV (real-time guidance) and after BMV (debriefing) improves performance. Feedback during bedside resuscitations may reduce delayed and interrupted BMV, but requires expert clinician-educators. Mobile health (mHealth) technology could enable implementation and evaluation of feedback strategies at the bedside in LMICs. The overall goal of this study is to reduce newborn mortality by improving BMV in LMICs through bedside feedback using an innovative mHealth application called LIVEBORN. The specific aims of this study are to 1) develop LIVEBORN, an mHealth application to improve BMV, 2) design and evaluate feasibility of feedback strategies for LIVEBORN, and 3) evaluate effectiveness of LIVEBORN in a hybrid, randomized trial. This proposal will take place in 10 health facilities in Kinshasa, Democratic Republic of the Congo (DRC) with midwives. LIVEBORN will identify depressed newborns using heart rate from a new heart rate monitor and data on provider action’s entered by an observer. After comparing actions to recommended care, LIVEBORN will deliver real-time guidance and support debriefing. LIVEBORN will be developed through a scientifically rigorous process involving formative research, technical development and usability testing. Integrated mHealth strategies for feedback with LIVEBORN (one for real-time guidance and one for debriefing) will be designed in collaboration with Congolese midwives from two facilities using trials of improved practices. The final strategies will be evaluated in a 3-month feasibility test in preparation for a hybrid, randomized trial. In a hybrid, randomized trial, eight facilities will be cluster randomized to real-time guidance or debriefing with LIVEBORN. After a period of baseline data collection, midwives will implement their assigned feedback strategy with LIVEBORN. The effectiveness of feedback with LIVEBORN on BMV will be evaluated comparing baseline and intervention data. If feedback with LIVEBORN is effective, the relative effectiveness of real-time guidance versus debriefing will be evaluated. The primary outcome will be the time to initiation of BMV. Secondary outcomes will be interrupted BMV and 24-hour newborn mortality. Feasibility and acceptability of feedback with LIVEBORN will be evaluated using a mixed methods approach. This study will be executed by a strong collaboration of five institutions: the University of North Carolina at Chapel Hill (UNC), the Kinshasa School of Public Health (KSPH) in the DRC, Laerdal Global Health, RTI International and Jhpiego. KSPH’s capacity to conduct mHealth research will be strengthened through the development of an mHealth Implementation Science course and establishment of a KSPH-UNC Implementation Science Core.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Through novel deconvolution approaches for bulk RNA sequencing analysis, we identified two tumor-intrinsic subtypes of PDAC (basal and classical) that we have confirmed, are robust, replicable, prognostic and predictive of treatment response. We found that the basal subtype is consistently associated with poor outcome and have shown, through analysis of two clinical trials, that patients with basal subtype tumors do not respond to the 1st-line therapy FOLFIRINOX. These results strongly support the idea that molecular subtypes may be used to select treatment. Given the impact of our tumor-intrinsic subtypes on therapy response, we developed a single sample classifier, PurIST, that is now a CLIA certified assay and being evaluated as an integral marker for treatment selection in a clinical trial. In parallel, we developed a de novo approach, DECODER to deconvolve bulk tumors into compartments that allows us to determine tumor and TME specific characteristics in patients. Using the deconvolution approaches that led us to identify tumor-intrinsic subtypes, we have found two types of PDAC stroma: activated, and normal where patients with activated stroma have shorter survival. We have shown that CAFs are the contributory cells in activated stroma. Patients can be found to have a combination of tumor/stroma subtypes and the combinations have different impacts on outcome, suggesting that it is critically important to understand tumor-stroma interactions and how they affect treatment response. Similarly, we find that i/myCAF may differentially educate basal vs. classical subtype lines Our findings provide strong support for our central hypothesis that CAFs and tumor cells have interactions that together may alter tumor progression and response, making it critical that we understand the heterogeneity of the stroma, and specifically CAFs, their interaction with the tumor, for tumor-stroma context specific treatment response. Our team is uniquely positioned to comprehensively characterize CAF/NAF-tumor heterogeneity and interactions, response to treatment, and develop an integrative CAF-tumor subtype classifier to predict treatment response of patients in standard of care, stroma and immune modulating trials.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Vascular anastomosis is an important surgical technique whereby closely spaced stitches are used to connect blood vessels. This process is used frequently in organ transplantation, trauma repairs, and cardiovascular surgery. However, vascular anastomosis is time consuming and associated with serious complications and long recovery times. Use of fibrin glue in anastomosis has led to improved surgical outcomes and shorter operating times, but these glues are frequently impractical as their concentrated formulations create high-density gels with short working times, low cellular infiltration, and cold storage limitations. We have developed fibrin-based nanoparticles (FBNs) which we have used to deliver growth factors and promote healing in vivo. Unlike traditional fibrin glues, FBNs are pre-polymerized and use physiologically relevant fibrin/thrombin concentrations. Owing to their colloidal structure and the aforementioned properties, FBNs exhibit tunable gelation, increased cellular infiltration, room temperature storage, and enhanced drug delivery capabilities – including of fibroblast growth factor 2 (FGF2), a known stimulant of vascular repair. The objective of this proposal is the optimization, characterization and in vivo analysis of paintable and patch formulations of an FBN surgical sealant with tunable growth factor release. It is expected that these FBN sealants will demonstrate better functionality than current fibrin glues, with the benefit of longer work time, extended stability at room temperature, targeted growth factor delivery, and improved healing outcomes. Aim 1 will optimize the formulation of patch and flowable FBN glues. This will occur through characterization of the effects of changes in FBN concentration, thrombin concentration, and cofactor used, on the mechanics and functionality of the glues. Testing will probe polymerization and degradation dynamics, structure, mechanical properties, sealant ability, and safety profile. Modalities used will include rheometry, microscopy, mechanical testing, and novel ex vivo and microfluidic assays. Aim 2 will optimize the loading/release of FGF2 by FBNs and characterize the effects of FGF2-FBN sealants on vascular wound healing in vitro. FGF2 loading efficiency and release characteristics of FBN sealants will be determined and compared to high-density bulk fibrin glues. Endothelial and fibroblast scratch tests and wound closure assays will be used to assess healing outcomes in vitro. Aim 3 will compare FBN formulations (gel and patch; unloaded and loaded with FGF2) to current fibrin glues using an in vivo leporine model of carotid artery anastomosis. Angiography will be use to characterize vascular morphology and histology of sampled tissues will be used to evaluate signs of healing, restenosis, and hyperplasia. This proposal’s use of FBNs will lead to a novel surgical sealant with improved work time and tunable drug delivery profiles that boasts superior wound healing – allowing for faster operating times, fewer complications, and improved recovery. This technology will also increase the accessibility of surgical glues by removing cold-chain requirements, opening up their use in varied global health applications. Training in the associated research techniques and clinical skills required for this project will contribute to the success of the applicant during the next phases of her career on the path to becoming an independent physician scientist.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY Platelets are a blood component critical to hemostasis, i.e. the prevention of blood loss at sites of vascular injury. For hemostasis, platelets must activate their primary integrin (aIIbb3) through the small GTPases Rap1 and downstream protein Talin. Integrin activation is essential to platelet adhesion and aggregation. Activated platelets can also release their granule contents and undergo phospholipid scrambling to support the formation of a stable hemostatic plug. The mechanisms of platelet activation must be tightly controlled as hypoactive platelets cause prolonged bleeding while hyperactive platelets are associated with thrombotic disease. Platelets which undergo phospholipid scrambling expose a negatively charged phospholipid, phosphatidylserine (PtdSer), to their outer membrane surface. These so called procoagulant platelets recruit and activate coagulation factors resulting in the generation of thrombin and the formation of fibrin, an integral component of stable hemostatic plugs. Loss of PtdSer exposure or coagulation factors results in bleeding complications; conversely, excessive PtdSer exposure is associated with increased thrombotic risk. Despite the negative outcomes associated with dysregulation of PtdSer, there exists a gap in knowledge on the mechanisms regulating PtdSer exposure following platelet activation. The small GTPase Rap1 is a well-established regulator of platelet integrin activation and aggregation. Loss of Rap1 in vitro also results in decreased PtdSer exposure; however, the mechanism and in vivo significance of Rap1-dependent PtdSer exposure are unknown. Interestingly, Rho family GTPases (RhoA/Rac/Cdc42) also modulate PtdSer exposure in platelets, and studies in multiple cell types, including platelets, have demonstrated crosstalk between Rap1 and Rho GTPases. Therefore, I hypothesize that Rap1-dependent PtdSer exposure occurs through crosstalk with Rho family small GTPases, and that Rap1-dependent procoagulant activity is critical during hemostatic plug formation. Preliminary data supports my hypothesis as inhibition of RhoA signaling leads to increased PtdSer exposure in platelets lacking both Rap1 isoforms, Rap1A and Rap1B. I have also established a novel imaging platform which allows for the study of platelet-dependent procoagulant response during hemostasis in vivo. In aim 1 I will characterize the crosstalk between Rap1 and Rho Family GTPases during platelet activation leading to PtdSer exposure. In aim 2 I will use my novel 4-D imaging model of hemostasis to characterize how Rap1 signaling affects platelet- coagulation interplay in vivo. This work will expand our understanding of the regulation of Rap1 mediated platelet PtdSer exposure and its consequences in vivo.

NIH Research Projects · FY 2025 · 2022-09

Project Summary In this new and unfunded study, we will capitalize on the lessons from the past 15 years of psychiatric genomic. Based on these lessons, we propose an exceptionally novel and important set of aims to further knowledge of the genetic architecture of mental illness. We propose to perform whole-exome sequencing and SNP-array genotyping on >150,000 cases with severe psychiatric disorders along with a similar number of controls. It will be large, transdiagnostic, based on patients seen in clinical psychiatry, and comprehensively analyze ultra-rare exonic, rare copy number, and common variation. Because assay costs are prohibitive (on the order of $US 80 million), we are partnering with Regeneron Genomics Center (RGC) that will conduct all genomic assays. NIMH funding is within the $500K direct cost cap at each site. We will: (1) Acquire samples with clinically severe psychiatric disorders. Cases will have lifetime diagnoses of schizophrenia (SCZ), schizoaffective disorder (SAD), bipolar I disorder (BD1), or severe major depressive disorder (sevMDD). Roles: UNC is responsible for data coordination; the sampling sites are ISMMS (the Americas and East Asia) and Cardiff (Europe, Africa, and South Asia) and each will collate samples (i.e., MTAs, ethical approvals, individual consent, harmonize phenotypes, QC DNA). Phase 1 (Years 1-2) will focus on existing samples (N=100K cases). Phase 2 (Years 1-4) will focus on obtaining new samples (N=50K cases), and will enable colleagues from low-income countries to obtain genetic data that would otherwise be impossible. This will help those investigators and greatly increase diversity in genomics research. 2) Genomic assays (Years 1-4). Samples will be sent to RGC in batches from ISMMS and Cardiff. RGC will generate whole exome sequencing and SNP array data. UNC and RGC will jointly conduct alignment, QC, variant calling (SNVs, indels, SVs), and array processing (common SNPs, imputation and CNVs). QC includes assessment of multiple biases and comparison to independent datasets. Deliverable: analysis-ready data frames for rare exonic, rare CNV, and common genetic variation. 3) Analysis for substantive scientific aims. Briefly, the main analytical themes are to identify genetic variation associated with: (a) severe mental illness, (b) specific disorders, and (c) cross-cutting clinical features (e.g., psychosis, treatment resistance, mania, ID). All analyses will be conducted using robust methods/bias control, formally compared to relevant prior studies, and evaluate the impact of all types of measured genetic variation across diverse genetic ancestries. 4) Data sharing will align with NIMH policies via the NIMH Data Archive. Successful completion of the proposed work will markedly increase the number of genes pinpointed by burdens of rare coding variation, rare CNVs, as well as less specific GWAS associations–we will markedly increase knowledge of the genetic architectures of these critically important and burdensome disorders.