Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2026 · 2024-02

Medically underserved populations are disproportionately affected by screen-preventable cancers, such as colorectal cancer (CRC). Entities such as the United States Preventive Services Task Force and the American Cancer Society recommend fecal immunochemical testing (FIT) as an effective home-based method for CRC screening. Annual FIT, when followed by a diagnostic colonoscopy for a positive result, can reduce CRC mortality on a population level. CRC screening is traditionally delivered as part of a medical visit. However, due to short visit times and competing demands among providers and patients alike, patients, even those who regularly attend medical visits, may miss out on screening, putting them at a higher risk of serious disease. FITs can be easily distributed in non-traditional healthcare settings and can help bridge the gap in care for underserved populations. Community pharmacies hold promise as a site for CRC screening because of their accessibility and pharmacists’ expanding role in delivering preventive services. Based on our formative work demonstrating the acceptability and feasibility, as well as pilot effectiveness, of FIT delivery in pharmacies, our objective is to evaluate the effectiveness and implementation of a pharmacy-based FIT intervention called PharmFIT. Through collaboration with community partners such as the North Carolina Network Consortium (NCNC), the Washington, Wyoming, Alaska, Montana, and Idaho Practice Research Network (WPRN) and the Community Pharmacy Enhanced Services Network (CPESN), our aims are to: 1) evaluate the impact, 2) assess the implementation, and 3) estimate the costs of the PharmFIT intervention. We will conduct a randomized controlled trial to assess the impact of PharmFIT on CRC screening rates in eligible primary care patients who are not up to date. Patients will be individually randomized to a usual care arm or a PharmFIT arm and we will determine whether there are statistically significant differences in CRC screening rates. Concurrently, we will use a mixed-method approach to assess a range of preliminary implementation outcomes, identify barriers and facilitators (i.e., determinants) of those outcomes, and identify implementation strategies to support future PharmFIT implementation to be explored in future research. Finally, we will calculate the cost of PharmFIT using collaborative process flow mapping to inform micro-costing and budget impact analysis. We hypothesize that the PharmFIT intervention will increase CRC screening uptake over usual care by providing better access and more opportunities for preventive services through a collaborative pharmacy/primary care approach.

NIH Research Projects · FY 2025 · 2024-02

Summary UNC-IMSD The long-term goal of the Initiative for Maximizing Student Development at the University of North Carolina at Chapel Hill (UNC IMSD) is to contribute to diversifying the leadership of biomedical science. We work towards this goal through an integrated suite of synergistic junior scientist development programs focused on biomedical PhD students from groups under-represented in the sciences due to historical exclusion. Our approach is a comprehensive, personalized, start-to-finish program led by a team of expert PhD-trained Directors and dedicated faculty who work to support the scientific, personal, and professional development of UNC biomedical PhD students from under-represented groups (URGs). We provide activities targeted to empower URG students and enhance both scientific identity and self-efficacy at key transition points in their graduate training to increase the likelihood that they persist to degree completion and in their chosen scientific careers. UNC IMSD is embedded in the UNC Office of Graduate Education, an institutionally-funded administrative unit that coordinates development, recruitment, and extensive support for 15 PhD programs. Leadership in this Office consists of multiple PhD-trained program Directors, including the two Directors of IMSD, whose full-time roles are to facilitate student success and to inspire continual improvement in graduate education practices in collaboration with the faculty PIs. The substantial resources devoted to graduate education at UNC through this Office provide numerous advantages to IMSD participants and members: i) ready access to expert advisors who have been through similar training , ii) a large and supportive community of peers from similar backgrounds and experiences, iii) individual academic support at key transitions (e.g. coursework, critical literature analysis, rotation and thesis lab selection, qualifying exams), and iv) customized professional development opportunities. Since 2006 the IMSD has financially supported 196 participants; 60 are still enrolled and on track to graduate, and 107 have graduated with a PhD (>85% retention). Participants published 308 peer-reviewed publications, 137 as first authors. Moreover, during this period an additional 64 graduate students were affiliated members of the IMSD community and benefitted from the program. As an R25-supported program since 1996, we institutionalized and broadly disseminated multiple successful innovations developed over many years. These elements benefit all graduate students – from under- represented and well-represented groups – and include faculty mentor training and a dedicated wellness counselor. We also engaged in scholarship of graduate admissions, graduate training, and workforce diversity; our studies have been published and have had impacts beyond UNC. In the proposed next phase of UNC IMSD as a T32, we have the following core Objectives: 1) develop and recruit high potential students from URGs, 2) facilitate academic and degree progress to PhD completion, 3) foster a community of thriving future scientific leaders, and 4) promote an inclusive training environment and cultural change.

NIH Research Projects · FY 2026 · 2024-02

SUMMARY / ABSTRACT Multiple clinical trials have shown that combining anti-estrogen therapy with CDK4/6 inhibitor therapy improves progression-free survival in ER+/HER2- breast cancer patients. However, many patients are resistant to CDK4/6 inhibitors or acquire resistance within the first few months of treatment. Interestingly, histological staining of treated tumors reveals a small but consistent population of cells in the proliferative phase of the cell cycle—a phenomenon we refer to as fractional resistance. It is critical to understand the mechanistic basis for fractional resistance because the precise fraction of proliferating tumor cells (e.g., 2% versus 15%) is a strong predictor of patient outcomes. Recent single-cell studies have shown that individual cells can take distinct trajectories, or “paths”, through the cell cycle that are defined by a unique combination of molecular states over time. We hypothesize that fractional resistance in breast tumors occurs because cells can take multiple paths through the cell cycle, but only some of these paths are sensitive to CDK4/6 inhibitors. To test this hypothesis, we have developed a powerful new method to profile >50 cell cycle regulator proteins in single cells. By linking single-cell states together, computationally, this approach has revealed how individual cells take alternate paths to evade cell cycle-targeted therapy. In Aim 1, we will induce fractional resistance in a panel of breast tumor models by applying increasing doses of CDK4/6 inhibitors and endocrine therapy to gradually eliminate subpopulations of proliferating tumor cells, thereby defining the range of molecular states cells can reach under drug treatment. Preliminary work suggests that resistant cells must take a particular set of paths at the G1/S transition characterized by high ratios of cyclins (e.g., cyclin D/E) to CDK inhibitors (e.g., p21/p27); altered CDT1 expression; and elevated E2F1 levels. We will validate these predictions through time-lapse microscopy. Aim 2 will take an unbiased approach to determine changes to the global protein landscape in response to increasing doses of endocrine/CDK4/6 inhibitor therapy. Using deep, quantitative proteomics that combines massive offline peptide fractionation with tandem mass tag labeling, we will determine changes to the protein landscape in response to CDK4/6 inhibition; validate these changes biochemically and by imaging; and determine the impact of the molecular state changes on sensitivity or resistance to endocrine/CDK4/6 inhibitor therapy. New preliminary results show multiple candidate regulators (e.g., CDKN3, UHRF1) that are cell cycle - dependent or expressed in fractionally resistant subpopulations. Aim 3 will identify resistant paths in resected human tumors—a technique developed by our surgical team. In addition, we will test the fractional resistance hypothesis in three ER+/HER2- PDX models to assess the extent of fractional resistance in long term, physiologically relevant model. Overall, this project will provide new knowledge in how heterogeneity in cell cycle behaviors leads to fractional drug resistance in ER+/HER2- tumors; identify novel resistance factors; and pave the way for the development of next-generation biomarkers that capture the unique cell cycle behaviors and drug sensitivities in other cancer types.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Innovative medical technologies can improve health and increase longevity for older adults with chronic disease and multimorbidity, yet new data are needed to promote their adoption and effective use in real-world settings. For example, nearly a quarter of all adults ≥65 years old in the US have diabetes. Technology-based approaches to diabetes management, such as continuous glucose monitoring (CGM), can improve clinical outcomes and quality of life in this age group, in addition to preventing dangerous episodes of hypoglycemia. Despite the potential benefits, CGM remains underutilized among older adults compared to younger adults. My career objective is to generate the scientific evidence needed to expand and improve the delivery of guidelines-aligned care for the expanding population of older adults with diabetes, and particularly with respect to rapidly emerging technology such as CGM. To this end, analyzing healthcare data (e.g., medical record data, insurance claims) can directly complement the existing evidence, and particularly randomized clinical trial (RCT) data, by revealing how therapies work in routine care, over longer durations of time, and in populations that are typically underrepresented in those RCTs (e.g., those ≥65 years and subgroups with multimorbidity, frailty, and cognitive impairment). Economic evaluation can further elucidate the broader implications of scaling uptake and sustained use of evidence-based, technologic therapies in older adult populations. My goal for this K01 award is thus to acquire rigorous training under interdisciplinary mentorship to develop (1) an understanding of the strengths and limitations of healthcare data sources and linkage solutions, (2) the capacity to apply advanced causal methods to analyze linked healthcare data, and (3) experience with economic evaluation. My proposal has exciting, high-impact training opportunities to address three fundamental gaps in the literature, including sparse research characterizing (1) patterns of CGM use among older adults and (2) the clinical effects of long-term use in such real-world settings, as well as a dearth of data on the (3) economic impacts of scaling CGM in this age group. I will (1) use a linked, real-world healthcare database to characterize subgroups of older adults with insulin-requiring diabetes who do and do not use CGM; (2) model the clinical effects of long-term CGM use on outcomes that are relevant for both endocrinology and geriatric medicine, and across key subgroups (e.g., ≥80 years, with cognitive impairment, multimorbidity, and recurrent hypoglycemia); and (3) evaluate the cost-effectiveness of scaling CGM use in this population. Completion of my aims will generate new evidence for CGM use in older adults that is relevant to patients, providers, and payers— a critical step to increase widespread uptake. By the end of the award, I will be positioned as a future leader in aging research with expertise to evaluate the real-world clinical and economic impacts of evidence-based technologies for older adults. I will use these skills to bridge innovations in diabetes care with the population of geriatric patients who stand to benefit most yet remain underrepresented in RCTs.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY/ABSTRACT Breast reconstruction is a critical component of breast cancer treatment because it restores quality of life and body image after mastectomy. It has become more common in the past 25 years, growing from 3 to 40% of mastectomy cases. However, Spanish-speaking Latina women who undergo mastectomy are the least likely to undergo reconstruction, meet with a reconstructive surgeon, or receive enough information about reconstruction. Those who do not have reconstruction experience the highest rates of dissatisfaction and regret about their decisions. Hispanic and Latino people are the largest minority population in the U.S. (19% of the population). Most (73%) primarily speak Spanish at home, and many (15%) feel uncomfortable with English. Persons diagnosed with cancer who primarily speak Spanish and identify as Latin American are less likely to receive guideline-concordant cancer treatments. Shared decision making and decision support can improve treatment decisions that are preference sensitive (the right choice depends on the person’s preferences). They result in higher patient knowledge and greater concordance between preferences and treatment. Unfortunately, most decision aids in the U.S. are in English and developed with few Latinx people. We have developed a decision aid for breast reconstruction called BREASTChoice, with proven efficacy and effectiveness in two randomized controlled trials. BREASTChoice was developed with extensive stakeholder engagement and includes a risk prediction model for personalized estimation of complication risk. Patients and clinicians are highly supportive of BREASTChoice and have asked for a Spanish language version that can be shared easily. Aim 1. Adapt BREASTChoice for Spanish-speaking Latina breast cancer survivors. We will interview Spanish-speaking Latina breast cancer survivors and other community-based key informants to culturally adapt and translate BREASTChoice. Our community partners, Living Beyond Breast Cancer and Latinas Contra Cancer, will help us identify participants. The adapted tool will be programmed as a mobile- friendly website. Aim 2. Assess usability and efficacy of the adapted BREASTChoice among Spanish-speaking Latina breast cancer survivors. We will conduct a pre-post evaluation among Spanish-speaking breast cancer survivors (N=50), who will use the tool and complete surveys before and after using it. Surveys will assess knowledge about breast reconstruction and the adapted tool’s usability. This study will result in the first culturally and linguistically relevant decision aid for breast reconstruction for Spanish speaking Latina breast cancer survivors. The decision aid’s format as a mobile friendly website will position it for wider dissemination and implementation because it will be portable and adaptable. The approach we use in this study will be widely applicable to other decision aids, so that decision support can reach Spanish-speaking Latinx populations.

NIH Research Projects · FY 2026 · 2024-02

Abstract We and others have shown that basal tumors, which comprise <20% of PDAC tumors, are resistant to the first- line chemotherapy regimen FOLFIRINOX. We show that basal tumors are significantly enriched in receptor tyrosine kinases, including EGFR. While EGFR inhibitors have been consistently disappointing in PDAC, our analysis of two clinical trials where patients were treated with gemcitabine in combination with an EGFR inhibitor, shows that the negative results are very likely driven by the previous lack of knowledge of subtypes. In both clinical trials, patients with basal tumors significantly benefit from EGFR inhibitor treatment. It is noteworthy that the survival of patients with basal subtype tumors is significantly worse than patients with classical tumors. We show that, for the very first time, basal patients who receive anti-EGFR therapy benefit from EGFR inhibitor therapy (patients with classical subtype tumors do not), and can achieve equivalent outcome to classical patients. Our findings strongly support that the positive subgroup analysis of basal patients is explained by the higher EGFR kinase expression found in basal tumors. These data strongly support our hypothesis that targeting subtype-specific kinases will be key to promising precision oncology approaches for kinase inhibitors in PDAC. We propose a multi-disciplinary team of a translational researcher (Yeh), kinome expert (Johnson), and medical oncologist (Somasundaram), that takes the next step in determining if EGFR inhibitors should be repurposed for patients with basal subtype tumors, and leverages a transformative high sensitivity method of defining the kinome in low input samples such as biopsies, to identify targeted approaches specifically for basal subtype tumors.

NIH Research Projects · FY 2026 · 2024-02

Project Summary/Abstract Seizures, a reflection of extreme excitatory-inhibitory imbalance, are a common neurologic comorbidity in individuals with intellectual and developmental disabilities (IDDs) and are even more prevalent in syndromic IDDs. Individuals with Angelman syndrome (AS), a severe neurodevelopmental disorder, are particularly impacted, as over 90% of AS individuals experience seizures, in addition to other symptoms such as severe intellectual disability, motor deficits, lack of speech, and sleep disruption. The recurrent seizures affecting AS individuals are usually difficult to treat and sometimes deadly, yet clinical efforts to alleviate seizure burden have been stymied by a lack of insight into ictogenic and epileptogenic mechanisms. I hypothesize that improved understanding of the circuitry underlying seizures in AS model mice can be leveraged toward the development of improved anti-epileptic therapies to address this unmet clinical need. This project builds upon previous work from our laboratory demonstrating enhanced epileptogenesis in AS model mice that have undergone seizure kindling, a process whereby repeated seizure inductions alter neural circuitry to increase seizure susceptibility. Prominent neuropathology arises post-kindling in AS mice in the dentate gyrus of the hippocampus, making this region a key mechanistic candidate. Moreover, we have found that deletion of Ube3a, the causative gene in AS, from GABAergic neurons expressing parvalbumin (PV), but not somatostatin or vasoactive intestinal peptide, drives seizure phenotypes. In the proposed project, I aim to elucidate the role of PV+ neurons in the seizure susceptibility of AS model mice. Accordingly, in Aim 1, I will perform whole-cell slice electrophysiology to determine how the intrinsic firing properties of PV+ neurons in the dentate gyrus differ between AS and wild-type mice, and whether these properties favor hyperexcitability in AS model mice following seizure kindling. In Aim 2, I will selectively reinstate Ube3a in PV+ cells to assess whether this confers resilience to kindling and prevents hippocampal histopathology. These experiments will provide important information at the cellular and circuit levels regarding how loss of Ube3a promotes seizure, potentially inspiring new treatments for AS and other epilepsy disorders. In addition to providing novel insights into circuit mechanisms of hyperexcitability, this project will provide the applicant with excellent training in slice electrophysiology (Philpot and Manis), confocal microscopy and image analysis (Itano), and the care of pediatric epilepsy patients (Yang), that will prepare him well for a career as a physician-scientist.

NIH Research Projects · FY 2026 · 2024-02

Title: Computational Neuroimaging MRI for Studying Early Brain Development with Autism Due to the absence of early biomarkers of autism, diagnosis must rely on behavioral observations long after birth, leading to missed opportunities for early intervention. Thus, it is of great importance to detect autism earlier in life for better intervention. The increasing availability of large-scale multimodal infant neuroimaging data (structural, functional and diffusion MRIs), e.g., Multi-visit Advanced Pediatric (MAP) Brain Imaging Study, Baby Connectome Project (BCP), and National Database for Autism Research (NDAR), affords unprecedented opportunities for precise charting of dynamic early brain developmental trajectories of autism, potentially providing important clues relevant to early detection of autism. Our hypothesis is that accurate characterization (segmentation, parcellation, multimodal neuroimaging measurements) of infant brain MRIs acquired from multiple centers will provide important insights into the origins and aberrant growth trajectories of autism and help identify potential imaging-based biomarkers for the early diagnosis of autism. To fully benefit from the large-scale multi-center infant neuroimaging data, a major barrier is the critical lack of computational characterization tools for accurate and robust processing of the challenging infant MRIs, which typically exhibit dynamic and extremely low tissue contrast, and large data heterogeneity. Building upon our existing research, this project aims to create and disseminate novel computational characterization tools that will enable accurate segmentation of cerebrum, cerebellum, and subcortical structures, parcellation, and measurements of infant brain structural, functional and diffusion MRIs from multiple imaging centers, and to further identify imaging-based biomarkers for early diagnosis of at-risk infants. To achieve our goal, we propose 4 specific aims. We will develop a novel contrast-enhancement network to increase the tissue contrast and a novel prior-guided transformer for enhanced and harmonized cortical and subcortical segmentation (Aim 1). We will propose a novel joint super resolution and tissue segmentation for cerebellum, aiming to achieve fine-grained segmentation results in an isotropic 0.4mm (or higher) space (Aim 2). Based on essential semantic features from segmentation maps in Aims 1 and 2, as well as contextual folding features from cortical surfaces, we will further develop a novel hybrid volume-surface parcellation framework trained on an innovatively augmented large-scale and diverse dataset (Aim 3). Finally, we develop a joint clinical scores regression and diagnosis model with attention mechanisms using multimodal features, including volumetric features from T1w and T2w scans, segmentation and parcellation maps (Aims 1-3), surface features from subcortical structures, cerebral and cerebellar cortex, and connectivity features from fMRI and dMRI, aiming to achieve high accuracy and interpretability in autism diagnosis (Aim 4). We will freely release our tools and all our processed data, to NDAR.

NIH Research Projects · FY 2026 · 2024-02

ABSTRACT Antibody-based therapeutics have grown rapidly over the past two decades. However, many challenges limiting the success of this drug class persist. First, the attrition rate remains high in antibody drug development. Second, many antibody therapeutics exhibit only transient pharmacologic action. As a result, antibody therapeutics benefit only a subset of patients. A key driver of these issues is our poor understanding of antibody pharmacology in vivo across biological contexts. Our research has revealed that antibody pharmacology is highly dependent on the physical and biological contexts of the biological environments where their actions occur. Methods and tools that integrate tissue physiology, disease pathology, and immune status into the characterization of antibody pharmacology are sorely needed. Leveraging multi-dimensional spatiotemporal data, we have developed quantitative systems pharmacology (QSP) models and experimental tools that allow us to investigate many facets of antibody context-dependent pharmacology, including tissue-specific pharmacokinetics (PK) and pharmacodynamics (PD), disease pathology, antibody-mediated immune dynamics, and cell-cell interactions. Our work has yielded insights for antibody drug development and therapeutic application. In this MIRA, we propose to apply and refine these experimental and mathematical tools to continue our long-standing efforts to address critical issues surrounding antibody context-specific pharmacology. Specifically, we will continue our work in 1) measuring and modeling antibody tissue PK and target engagement in living animals; 2) modeling antibody-mediated cell-cell contact and interactions across biological contexts; and 3) determining the roles of the constant region (Fc) and effector function in soluble target neutralization. In project 1, our lab has developed a proximity-based bioluminescence resonance energy transfer (BRET) imaging approach. This BRET imaging technology can longitudinally measure antibody exposure and target engagement within tissues of living animals, elucidating the physiological and biophysical factors that govern and restrict antibody−target binding processes in vivo. In project 2, we will apply our proximity-based cell-cell imaging tools, coupled with a multidimensional QSP model for cell-cell interactions, to uncover the biophysical principles of antibody-mediated cell-cell communication. In project 3, we will investigate the multifactorial aspects of antibody pharmacology for soluble target neutralization and highlight the critical but neglected roles of Fc variants and effector function. This project could uncover overlooked mechanisms that have restricted antibody therapeutic development against soluble targets and expand the field’s design space. Overall, our research will continue to address the fundamental challenges to efficient antibody development and optimal therapeutic application through the development of experimental tools and QSP models.

NIH Research Projects · FY 2026 · 2024-02

Abstract This goal of this project is to develop computer simulations of combination therapy for aggressive lymphomas, and apply them to understand past positive and negative trials, and to enable model-guided design of new regimens. Theoretical models of cancer drug response and resistance evolution have provided many conceptual insights, but models of a `representative' tumor have not been able to predict response distributions in heterogeneous human populations. We recently developed simulations which use clinically observed distributions of single-drug responses to predict distributions of multi-drug responses. These models have been validated by accurately predicting many trial results in solid cancers (9 FDA approvals), but our prior models were too simple to describe curative treatments because they lacked intra-tumor heterogeneity and kinetics. Here we propose conceptual and technical advances to model the complexity of curative therapies, using Diffuse Large B-Cell Lymphoma (DLBCL) as a case study. Our simulations of multi-drug response will consider inter-patient and intra-tumor heterogeneity, tolerability and dosage, treatment schedule and response kinetics, and drug interactions. We adopt the conceptual approach of population-pharmacokinetics, where each parameter has a distribution describing its variance among patients. We will apply this approach to tumor drug response, considering both intra-tumor and inter-patient variation. Parameter distributions are informed by our experimental data from clone tracing and liquid biopsies to quantify clonal heterogeneity and response kinetics, and digitization of decades of trial data. Aim 1 will analyze past and current trials of drug combinations in first- line DLBCL to test whether the clinical efficacy of drug combinations is predictable from single drug efficacy. Preliminary data shows the past 20 years of novel combination trials in first-line DLBCL confirm model accuracy, and we prospectively predicted the first success in 2 decades. This aim will produce predictive models that can help design future drug combinations. Aim 2 will investigate explanations for the negative result of trial that added a targeted therapy, ibrutinib, to standard chemotherapy. We will model the influence of tolerability and dose reductions, enrollment bias, and treatment schedule, comparing model outputs with real- world analyses of how these factors affect outcome. By understanding causes of trial failures this aim can help future trial designs to overcome these problems. In Aim 3 we will collaborate with the ECOG-ACRIN trial group to apply model-guided design to a trial of precision combination therapy in first-line DLBCL. The LymphoMatch trial aims to match 5 subtypes of DLBCL to 5 targeted therapies, combined with standard chemotherapy. We will use clinical data on single drug efficacies, combination tolerability, subtypes' prognoses, and accuracy of subtypes as biomarkers of drug sensitivities, to forecast trial results and so optimize design of regimens and endpoints. This research will deliver innovative multi-scale models of tumor heterogeneity, to solve challenges in the design of novel combination therapies and clinical trials that aim to cure more cases of lymphoma.

NIH Research Projects · FY 2025 · 2024-01

The longstanding paradigm in vaccine development has been the introduction of immunogens that culminates in generation of pathogen-binding antibodies (Ab). While the precise format of the antigen presentation differs, all vaccines rely on somatic hypermutation of immunoglobulin genes in B cells to generate Ab with greater affinity, avidity or anti-pathogen activity. Unfortunately, the natural variations in our immune system leads to highly variable Ab responses, both in magnitude of the induced Ab response and the breadth/potencies of the specific Ab clones generated, resulting in variable efficacy. Here, we seek to overcome these shortcomings by directly reprogramming circulating B cells to secrete specific potent Ab of interest, including stable integration of Ab-encoding transgenes into circulating B-cells. The resulting chimeric antibody reprogrammed (CAR) B cells can produce Ab with magnitude, affinity and effector functions that can all be controlled with molecular/genetic specificity. We refer to this as in vivo engineering of CAR-B cells. The key to realizing this vision is a delivery platform that can facilitate highly specific transduction of B- cells in vivo. We have engineered lentivirus (LV) vectors that effectively transduce circulating immune cells. Our first-generation system achieved highly specific transduction of circulating T-cells even at very low virus:cell ratio, and generated substantial CAR-T cells in the circulation and in tumor of immune-deficient NSG mice with highly aggressive BV173 lymphoma, leading to effective suppression of the tumor and prolonged survival. We have since improved upon our first-generation system with a second-generation LV system incorporating the Nipah virus fusion protein. Nipah LV (NLV), coupled with a proprietary combo of transduction enhancers (TE), efficiently and specifically transduced non-activated B-cells in human PBMCs. In pilot studies, a single injection of NLV+TE into NSG mice with circulating human PBMCs lead to effective transduction of >0.5% of all circulating B-cells. We have also demonstrated we can harness CRISPR/Cas9 to insert Ab transgene into a well conserved site in B cells, leading to functional B cells that can react to antigens. In this proposal, we seek to perform key enabling studies that substantiate our proposed CAR-B strategy. In Aim 1, we will engineer NLV that targets different B-cell populations, and identify the combination of NLV and TE that maximizes transduction of key human B-cells. We will then assess delivery of transgene encoding for a potent RSV neutralizing Ab (nAb) either by non-specific integration or site-specific integration. We will then advance into animal studies in Aim 2, where we will assess transduction efficiency and specificity in NSG mice infused with circulating human PBMCs. We will quantify for the levels of the nAb in serum over time, and assess protection against infection by respiratory syncytial virus (RSV). If successful, our work will advance an alternative strategy for achieving highly tunable immune protection in cases where specificity of the variable domain or Ab isotype is essential, or in individuals who do not respond to traditional vaccination.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT Artemisinin-based combination therapies (ACT) have been the bedrock of malaria treatment in Africa for more than a decade, spurring remarkable gains in malaria control, even as parasites tolerant to commonly used ACT partner drugs continue to circulate. In East Africa, parasites harboring mutations associated with resistance to the partner drug lumefantrine are selected through treatment and responsible for the majority of recurrent infections that arise starting 3-4 weeks following ACT. These post-treatment parasitemias likely contribute to the onward transmission of drug-resistant parasites to mosquitoes and further spread of antimalarial resistance. Now, parasites with partial artemisinin resistance that have emerged in Rwanda are spilling over into northwest Tanzania, a high malaria burden country, where 4% of the world’s malaria deaths occur. Mitigating the spread of ACT-resistant parasites across sub-Saharan Africa will require a better understanding of the transmission reservoir (who is transmitting drug-resistant parasites to mosquitoes) and ways to intervene to interrupt this transmission. While ACT and single low dose primaquine effectively clear those presenting with gametocytes, the parasite stages essential for transmission to mosquitoes, no current strategies target gametocytes that arise in the weeks following treatment. We hypothesize that gametocytes associated with post-ACT parasitemia are transmissible to mosquitoes and contribute to the spread of antimalarial resistance, even as most go undetected. This R21 proposal will determine the risk of transmission from PCR-positive individuals one month following ACT (Aim 1) and find whether drug resistant alleles that are selected during treatment are maintained through transmission (Aim 2). To accomplish these aims, we will leverage the capacity we have built over the last 5 years to characterize the infectious reservoir in Bagamoyo, Tanzania, where we have performed >600 mosquito feeding assays in asymptomatic parasite carriers, most with subpatent infection only detectable by PCR. Our team will screen >300 persons participating in a large triple ACT trial for recurrent parasitemia during weeks 3-8 of follow-up and conduct mosquito membrane feeding assays to measure their transmissibility to Anopheles gambiae and A. funestus, the primary malaria vectors in East Africa. We will sequence transmitted parasites that survive to the salivary gland sporozoite stage within mosquitoes and compare the frequency of key drug resistance alleles in these mosquitoes to blood collected from persons pre and post-treatment. We expect that the post-treatment period, 4-8 weeks after first-line ACT therapy, is indeed a vulnerable time for gametocytemia to arise without symptoms, and lead to transmission of P. falciparum parasites bearing kelch 13 and mdr1 mutations selected through treatment. However, we also expect that triple ACT therapies that reduce the rate of post-treatment parasitemia will be effective in preventing onward transmission of drug-resistant parasites. Findings from our study will inform the design of strategies that mitigate the spread of ACT resistance, buying time for new antimalarials to be rolled out.

NIH Research Projects · FY 2026 · 2024-01

NINDS supports summer research experiences (SREs) to attract and prepare undergraduate students for careers in neuroscience. For nearly 40 years, the Carolina Summer Fellows Program (CSFP) in the UNC Department of Pharmacology has offered SREs to many undergraduates, successfully helping retain these students in academic and STEM careers. About six years ago, we shifted the focus of the CSFP to provide research experiences, establish lifelong mentorship, and build support networks. Since then, we have trained over 50 students through the CSFP, with many going on to enroll in master's, doctoral, and medical programs nationwide. We have built a steady pipeline of talented fellows mainly through institutional partnerships. During this five-year period, the Department of Pharmacology has also expanded its faculty specializing in neuropharmacology and neurobiological diseases. Additionally, we see high interest in neuroscience research among our student applicants (currently >50 per year). However, since the CSFP has been a self-funded program, we have been unable to fully meet the demand in this area. Therefore, we propose establishing the CSFP-Neuro program in this application, which takes advantage of our faculty's expertise in neurological diseases and neuropharmacology at UNC, as well as our established partnerships with collaborating institutions. The program aims to provide undergraduates with high-quality SREs focusing on the neurobiology of disease, involving faculty from the Department of Pharmacology and the UNC Neuroscience Center. Each faculty and bench mentor will receive training in effective mentoring practices to improve communication, align expectations, and foster independence. Besides research, we plan to include scientific and career mentoring, career development activities, networking opportunities, and professional growth initiatives designed to attract students to neuroscience careers and prepare them for graduate school. Our program has a proven track record of implementing these career development activities to benefit students. To further support retention in neuroscience research careers, we also propose ongoing mentoring, professional development, and assistance with graduate school applications after the SRE. This comprehensive approach is intended to equip students with the tools, experience, and support necessary to succeed in neuroscience research careers.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT Chronic suppurative respiratory disease is characterized by persistent and recurrent otitis media, sinusitis, bronchitis as well as bronchiectasis. Genetic etiologies include inborn errors of immunity (IEI) with defective cellular and antibody-mediated responses and disorders of impaired mucociliary clearance like primary ciliary clearance (PCD). Early and accurate diagnosis of these genetic etiologies can lead to more effective prevention of recurring infections, functional decline, and structural respiratory tract damage. Prior studies conducted by the NIH-funded Genetic Disorders of Mucociliary Clearance Consortium (GDMCC) focused on characterizing patients presenting with suspected PCD. These led to better understanding of the clinical phenotype and contributed to the discovery and characterization of over 50 genes causing this disease. Those studies also showed that some patients presenting with signs and symptoms of PCD actually had milder or later onset manifestations of IEI identified genetically. This project seeks to complement the extensive PCD clinical, genetic and management experience of the GDMCC with the corresponding IEI expertise of the NIAID Sequencing and Primary Immune Deficiency Programs to test our central hypothesis that tiered clinical and genetic evaluation of patients with suppurative respiratory disease will identify people whose confirmed genetic diagnosis of PCD or IEI requires whole genome sequencing and expertise in variant significance resolution. Utilizing patients identified from a multicenter GDMCC suppurative respiratory disease protocol, we propose the following aims: Specific Aim 1: Assess the potential of Whole genome sequencing (WGS) for diagnosis of IEI or PCD in patients who have undergone a systematic evaluation for suppurative respiratory tract disease and have negative commercial IEI and PCD gene testing panels. Expertise in variant curation and significance resolution for gene variants not currently categorized as pathogenic or likely pathogenic will be utilized to confirm the genetic diagnoses. Specific Aim 2: Referral of GDMCC patients to the NIH Clinical Center for NIAID protocol directed clinical, laboratory, or genetic diagnostic evaluation will facilitate confirmation of genetic IEI diagnosis. Patients will have the opportunity to participate in NIAID protocols designed to more precisely define immune disorders at the NIH Clinical Center. Specific Aim 3: Referral of GDMCC IEI patients to the NIH Clinical Center for NIAID protocol directed biologic, gene correctional or transplant treatment will enhance therapeutic options. Patients diagnosed with a genetic IEI will have access to potential lifesaving therapeutic advances. The collaborative efforts between the multi-site GDMCC based at the University of North Carolina at Chapel Hill and the NIAID Centralized Sequencing Program, and Primary Immune Deficiency Program based at the NIH Clinical Center will lead to improved genetic diagnostic capabilities and optimization of management strategies for patients presenting with chronic suppurative respiratory disease.

NIH Research Projects · FY 2026 · 2024-01

Abstract HIV Pharmacology Data Repository (HIV PDR) Clinical pharmacology is paramount to developing new therapeutic strategies for HIV treatment, prevention, and cure that are needed to end the epidemic. Since its inception in 2001, the UNC Center for AIDS Research (CFAR) Clinical Pharmacology and Analytical Chemistry (CPAC) Core has led the field in the provision of high- quality clinical pharmacology services to HIV/AIDS researchers. The CPAC Core has consolidated >75,000 unique concentration vs time (CvT) datapoints resulting from >29,000 samples collected across 482 unique studies quantifying 72 drug molecules in 10 species and 14 anatomical compartments into a single searchable data portal - the HIV Pharmacology Data Repository (HIV PDR). Pooled CvT data can be easily extracted from the HIV PDR in machine readable output, making this data portal a valuable informatic tool for sharing and exploring antiretroviral pharmacokinetic data to accelerate biomedical research discovery. We seek to maximize the HIV PDR’s scientific impact by expanding and disseminating this resource through the following aims: 1) creating new tools to catalyze data expansion, 2) demonstrating utility by promoting scientific exploration, and 3) propagating use by engaging users and characterizing their Target Product Profile (TPP). To this end we propose to apply FAIR (Findable, Accessible, Interoperable, Reusable) principles to expand CvT data archives, promote exploration, and empirically inform downstream development of the HIV PDR. We will ensure the long- term sustainability of this valuable resource by creating advisory boards to oversee the development of a robust governance plan designed to ensure regulatory compliance while meeting user needs. Finally, in performing the proposed aims, we will pioneer a data-sharing service to assist HIV/AIDS researchers in meeting the recommendations provided within the 2023 NIH Policy for Data Management and Sharing.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY Breast reconstruction after mastectomy is critical to comprehensive breast cancer treatment because it can restore quality of life and body image, which are often impaired after mastectomy. However, many patients are not offered reconstruction, especially those who identify as Black or Hispanic/Latina. Many patients lack knowledge about breast reconstruction risks, and their choices about reconstruction often misalign with their preferences. We have developed a breast reconstruction decision aid called BREASTChoice and demonstrated its efficacy and effectiveness in two randomized controlled trials. Patients and clinicians found BREASTChoice highly usable but requested a version they could use on a mobile device and easily share with family and friends. Like most decision aids, BREASTChoice’s implementation has been limited to clinical settings and had limited uptake. For dissemination of decision support to be successful, it must make decision support available how, when, and where people want to use it. Dissemination through social media within online communities could broaden the reach of BREASTChoice and other decision tools. Most Americans use social media (81%) and use the internet for health information (72%). Even most older adults (60%) use the internet. Breast cancer survivors are especially engaged in social media and online communities. Our preliminary data suggest that breast cancer survivors would engage with BREASTChoice through social media. Guided by the Designing for Dissemination paradigm, and in partnership with 3 cancer survivor and two clinician organizations, we propose to adapt BREASTChoice and disseminate it through social media within online communities. Aim 1. Adapt BREASTChoice for mobile use and online dissemination. Guided by the ADAPT-ITT model, we will conduct iterative phases of testing in breast cancer survivors (n=45), qualitative interviews with topical experts (n=10), and website programming changes. Adaptations will be tracked using FRAME. Aim 2. Design and test social media messages for dissemination of BREASTChoice. Guided by Diffusion of Innovations Theory, we will engage key community partners regarding potential adoption and readiness for change; assess the social media dissemination context, including digital opinion leaders and platforms; and develop and test social media messages. Aim 3. Disseminate BREASTChoice to breast cancer survivors by leveraging social media. Using the adapted tool and tested messages, we will roll out a social media campaign over 12 months. We will test the campaign’s exposure, reach, and engagement with interrupted time series analysis, and BREASTChoice effectiveness (surgery choice, decision quality, decisional conflict, satisfaction with decisions) through a longitudinal user survey. Upon completion of these aims, we will have overcome a major scientific gap, the limited adoption of decision aids, and have developed a widely applicable approach to disseminating decision aids outside of clinical contexts.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT Following the global roll-out of rotavirus vaccines, attention is now focusing on vaccination strategies against norovirus, the next leading cause of viral gastroenteritis. Several norovirus vaccine candidates are in the pipeline, including an oral GI.1/GII.4 vaccine that will soon be tested in lactating women. The main goal of this vaccination strategy is to elicit norovirus-specific antibodies in breastmilk. However, very little is currently known about norovirus immunity in breastmilk. Epidemiologic studies do not consistently find breastfeeding to protect against norovirus. Also, no prior studies have fully characterized norovirus-specific antibodies in lactating women with natural norovirus infections to understand what might be attainable with postpartum vaccines. A better understanding of maternal immunity and how it protects infants against norovirus infections could guide novel postpartum vaccination strategies. In addition, pediatric vaccines are being developed. A parenteral GI.1/GII.4 norovirus vaccine is undergoing testing in children after encouraging Phase IIb results in adults. However, vaccine-elicited immunity in naïve infants may differ from that in adults, who have experienced multiple prior infections. For example, our group demonstrated that the dominant response to the parenteral GI.1/GII.4 vaccine in adults was boosting from a previous GII.4 infection. Further, we have demonstrated that natural norovirus infections in young children elicit a narrow antibody response—typically to the infecting genotype only— suggesting that an effective pediatric vaccine would need to include multiple norovirus genotypes. These differences in the immune response in adults vs. children suggest that understanding the unique immune stage of the infant is necessary to develop an effective pediatric vaccine. This project connects a robust field site with state-of-the-art immunological approaches to inform norovirus vaccination strategies to benefit children. We will enroll 120 Guatemalan breastfed infants with acute norovirus gastroenteritis and their mothers to determine if there is an association between norovirus-specific antibodies in breastmilk and the duration of norovirus infections in infants. Next, we will characterize and compare humoral immunity to norovirus in infants and mothers infected with the same norovirus strain. Finally, we will characterize the kinetics and breadth of norovirus-specific antibodies in breastmilk in lactating women with norovirus infections. We hypothesize that a) infants receiving breastmilk with higher levels of genotype-specific norovirus antibodies will have shorter norovirus infections as compared to infants receiving lower levels of these antibodies, b) infants will mount narrow antibody responses to norovirus as compared to (previously exposed) adults, and c) in lactating women with norovirus infections, norovirus-specific antibody responses in breastmilk will be broad and short-lived, similar to antibody responses to norovirus in another mucosal fluid, saliva. This project responds to the pressing need to inform norovirus vaccination strategies to benefit children, while adding to our basic understanding of immune protection at the maternal-child interface.

NIH Research Projects · FY 2026 · 2024-01

Chlamydia trachomatis is a well-recognized PID pathogen for which no vaccine is available. Women are at risk for reproductive sequelae when infection ascends into the upper reproductive tract, driving inflammatory processes that trigger immune pathology. However, the bacterial and/or host factors that govern ascension and subsequent pathology remain unknown. Our long-term goal is to identify biomarkers that identify women at risk for reproductive morbidity or correlate with protective immunity. We and others have established that susceptibility to human chlamydial genital infection is mediated by pathogen abundance, co-infection, oral contraceptives and cervicovaginal microbiota. The overall objective of this proposal is to identify cervical discriminators of asymptomatic, ascending chlamydial infection and endometritis in women. The central hypothesis is that overall risk for ascending infection is modulated by local host inflammatory responses, pathogen fitness and environmental factors mediated by the cervicovaginal microbiome. Our rationale is that identifying these factors will accelerate rational vaccine design and testing. In a pilot study of women at high risk for STIs, we demonstrated the potential for unbiased characterization of host, pathogen and microbiome interactions using transcriptomics and determined that a single cervical specimen was sufficient to detect clustered host transcriptional profiles reflecting microbiome differences and STI infection. Guided by strong preliminary data and leveraging ongoing NIH funded research, the following four specific aims will test our hypothesis: 1) Profile cervical immune response with respect to ascending infection and susceptibility to reinfection; 2) Profile chlamydial transcriptional activity associated with ascending infection; 3) Profile transcriptionally active microflora in Chlamydia-infected women to determine how these complex microbial communities modulate ascending infection; and 4) Integrate host, pathogen and microbiome responses to determine key pathways controlling outcome. The first aim will profile cervical inflammatory responses and immune cell populations from women with local infection or upper tract involvement in a cohort of highly-exposed women (TRAC2). We will transcriptionally profile the chlamydial strains causing their infections and use metatranscriptomics to characterize the contribution of the cervicovaginal microbiome. Finally, we will use robust biostatistical approaches to improve and expand our understanding of key chlamydia–host interactions that modulate infection outcomes, drive disease and establish protective immunity. The research proposed is innovative, in our opinion, because it will implement a comprehensive, non-biased approach to the identification of molecular biomarkers in a highly disease-relevant clinical population. The proposed research is significant because it is expected to translate directly to anti-chlamydial vaccine evaluation in humans and to have broad importance for diagnosis and for evaluation of novel therapeutics.

NIH Research Projects · FY 2025 · 2024-01

More than one in five adult Latinos in the United States has arthritis and Latinos experience greater joint pain and limitations than non-Latino Whites. Physical activity (PA) improves arthritis symptoms, yet Latinos are less likely to engage in any PA (50%) than non-Latino Whites (65%). Camine con Gusto (CCG), the Arthritis Foundation’s evidence-based 6-week Spanish-language version of Walk With Ease, is one of few PA interventions tested in Latinos. CCG was shown in a pre-post study to improve outcomes in 233 Latinos with arthritis. The Osteoarthritis Action Alliance developed an online portal for CCG with enhancements including weekly emails/texts, tracking/goal setting, and video/audio content based on feedback from Latinos and community-based organizations (CBOs) that offer CCG. Participants expressed a need for additional, more resource-intensive components to CCG that have yet to be implemented or tested. The overall goal of this R34 is to plan an assessor blinded community-based randomized optimization trial to test recommended additional components for CCG PA initiation and maintenance. To evaluate which additional components contribute meaningfully to increasing PA, we will use the Multiphase Optimization Strategy (MOST), an approach to developing highly efficient and scalable behavioral interventions that include only the most potent and cost- effective components. Using MOST, we will: 1) Determine the set of components (pedometer, CCG promotora, and maintenance promotora) when added to CCG that meaningfully increase daily steps (primary outcome) immediately post-CCG and 6- and 12-months post-CCG; 2) Evaluate the set of components, when added to CCG, that meaningfully improve secondary outcomes (i.e., arthritis symptoms, sleep quality) post-CCG and at 6- and 12-months; and 3) Evaluate the cost-effectiveness of the intervention components for scalability in CBOs serving Latinos with arthritis. We have worked closely with our Community Advisory Board (CAB) on this R34 proposal and will work with them on all aspects of the planning grant and proposed trial. Results from each aim will inform the decision on which set of components has the greatest impact on PA initiation and maintenance for the best cost. In this planning period, we will: 1) Use CAB input and behavioral theory to finalize the scientific details for the study; this will include refining CCG portal enhancements and additional intervention components (i.e., pedometer and promotoras), finalizing outcome measures, collection methods, the research and cost-effectiveness protocols and statistical analysis plan. 2) Use CAB input to develop the materials and application for the proposed trial; this will include recruitment, retention, and adherence materials and approaches; manual of operating procedures; data management plan; training manuals, and fidelity assessment plans; data safety and monitoring plan; and a detailed project timeline and budget for the future clinical trial. This proposed R34 will plan and prepare a well-powered randomized optimization trial aimed at reducing PA disparities among Latinos with arthritis, an understudied population in arthritis research.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY Our rapidly aging population remains at an exceptionally high risk of debilitating falls. This is especially concerning given the compounding effects of dementia. In particular, Dementia with Lewy Bodies (DLB) is uniquely associated with an increased prevalence of falls. Developing an improved mechanistic framework to understand balance impairment due to age aging and DLB to mitigate falls risk is at the heart of this proposal. The neuromechanics of standing and walking balance control, and thus the origins of balance impairment due to aging and DLB, are incredibly complex. Proactive responses precede the onset of a balance challenge via feedforward control to prepare the body to accommodate instability. Conversely, reactive responses follow the onset of a balance challenge and are compensatory, requiring rapid corrections to mitigate instability. The overarching scientific premise of this fellowship proposal is that aging negatively affects reactive balance responses, while the compounding effects of age and cognitive decline negatively affect proactive balance responses in people with DLB, thus increasing vulnerability to real-world balance challenges that can precipitate falls. This cross- sectional fellowship study will enroll 25 young adults, 25 older adults with clinically probable DLB, and 25 age- and sex- matched older adults. I will strategically combine quantitative motion capture, electromyography, dynamic in vivo ultrasound imaging, and wearable sensors with an innovative suite of standing and walking perturbation paradigms designed to emulate real-world balance challenges. Aim 1 will determine the effects of age and LBD on whole-body vulnerability to anticipated and unanticipated balance challenges during standing and walking. Aim 2 will characterize the local muscle neuromechanical determinants of reactive and proactive balance responses across these cohorts. Finally, with community- based translation as the ultimate goal of my scientific and professional development, Aim 3 will quantify the viability of wearable sensors to monitor and detect between-group differences in vulnerability to perturbations. This study will be the first to objectively quantify between-group differences in multi-scale characteristics of proactive and reactive balance responses using laboratory-based and wearable-sensor based outcomes designed to move my discoveries from the laboratory to the clinic and the community. This area of mechanistic and hypothesis-driven research has been severely understudied, but has significant and immediate potential to inform novel advances in diagnostics, rehabilitation, mobile monitoring, and wearable assistive technologies to mitigate falls.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT: Alcohol is one of the most widely used drugs by adolescents and is often consumed in a binge drinking pattern. Binge drinking alcohol during this critical developmental period can lead to lasting cognitive impairments for which there are few effective treatments. One domain that is disrupted following adolescent binge alcohol use is behavioral flexibility, defined as the ability to adapt behavior to a changing environment. The neurocircuitry underlying behavioral flexibility is complex but includes the medial prefrontal cortex (mPFC), the orbitofrontal cortex (OFC), and the anterior insula cortex (aIC), all regions that are among those perturbed by adolescent alcohol use. To better understand the neural mechanisms underlying adolescent alcohol induced cognitive impairments and to test novel treatment paradigms, our lab utilizes a rat model of adolescent binge alcohol exposure (adolescent intermittent exposure, or AIE). We have identified deficits in behavioral flexibility as measured by an attentional set shifting task in AIE-exposed rats compared to controls. Furthermore, we have recently identified AIE-induced changes in perineuronal nets (PNNs), a component of the extracellular matrix that preferentially forms around parvalbumin (PV) interneurons and modulates their activity. AIE was shown to increase PNN density in the mPFC and OFC, and a higher proportion of PV+ interneurons were surrounded by PNNs. Additionally, we have collected preliminary data suggesting that AIE leads to a higher proportion of PV+ interneurons surrounded by PNNs in the aIC. As these altered neurochemical markers are found in the mPFC, OFC, and aIC, they may underpin the observed deficits in behavioral flexibility following AIE. One potential strategy to improve deficits in behavioral flexibility is by using transcranial alternating current stimulation (tACS), a noninvasive neuromodulatory technique used in humans. tACS has been reverse translated and has been shown to restore flexible behavior in rats that have drug-induced impairments. Further, tACS is known to alter gamma oscillations, which are regulated by PV interneurons. However, it is not known whether tACS treatment will restore adolescent alcohol-induced deficits in behavioral flexibility. Additionally, it is currently unknown whether tACS can alter neurochemical markers. This F32 proposal will: 1) determine if tACS treatment normalizes PNN density in adult AIE-exposed rats; and 2) determine if tACS treatment restores alcohol-induced deficits in behavioral flexibility. Clinically, these studies will be impactful because they test the ability of tACS to ameliorate alcohol-induced deficits in behavioral flexibility, something known to be disrupted in humans following adolescent binge-alcohol consumption. The results generated from these Aims will lay the necessary groundwork for future investigations and grant proposals, such as a K99/R00 application. Completion of the proposed study will allow me to be trained in a highly clinically relevant technique (tACS), as well as expand my expertise in immunohistochemistry and behavior, and by doing so, prepare me to address important questions in the field of alcohol research as I progress towards my goal of becoming an independent investigator.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY Knowledge of how three-dimensional (3D) genome organization is linked to gene expression and chromatin state is paramount to understanding human health and disease. The cohesin complex is a major regulator of genome organization that dynamically extrudes DNA loops to bring together enhancers, promoters, and insulators. While the insulator protein CTCF is known to capture and stabilize an extruding cohesin complex, it is not clear how cohesin is stabilized at enhancers and promoters that lack CTCF. Furthermore, the causal role of cohesin-mediated loops in regulating gene expression is not understood. We recently discovered that cohesin variant complexes, composed of either STAG1 or STAG2 subunits and either PDS5A or PDS5B subunits, colocalize across the genome, yet have surprisingly distinct effects on gene expression. In this proposed study, we will answer the next set of questions about cohesin biology. We hypothesize that cohesin complexes at specific genomic sites have distinct biochemical properties and binding partners that mediate effects on transcription and chromatin state defined by histone modifications. To test these hypotheses, we are using an integrative approach that combines genetic, biochemical, genomic, and proteomic assays in embryonic stem cells and differentiated cells following genome editing, protein depletion, acute degradation, or inhibition with small molecules. These studies will 1) identify novel cohesin-interacting proteins using proteomic approaches, 2) investigate the properties of cohesin complexes at different genomic sites and with different binding partners, and 3) elucidate the role of cohesin-mediated DNA loops in regulation of transcription and chromatin state at genomic sites lacking CTCF. The proposed multi-disciplinary approach leverages our extensive experience in functional genomics and expands on our recent findings that demonstrated roles for cohesin subunits and cohesin regulators in DNA loop formation and transcription. This will contribute to our long-term goal of applying cutting-edge technologies to uncover the molecular mechanisms that define how 3D genome organization influences chromatin state and transcriptional control to govern cell identity. Completion of this project will fill critical gaps in knowledge about gene regulation that inform our understanding of human developmental disorders and cancers linked to epigenetic defects.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY Focused Ultrasound (FUS) is a powerful technology now FDA approved for several ablative brain therapies, and has demonstrated an impressive safety and efficacy profile for treating diseases such as essential tremor. Beyond ablation, early human studies demonstrate efficacy and safety of noninvasive microbubble-mediated blood-brain barrier (BBB) disruption with FUS for advanced therapeutic applications. In this high-risk, high- reward biomedical engineering project, we have assembled a uniquely qualified team to explore novel applications of FUS to improve Chimeric Antigen Receptor T cell (CAR-T) therapy for glioblastoma (GBM) treatment. CAR-Ts are a promising new treatment platform that allows directed targeting and killing of tumor cells. Locally administered CAR-Ts have demonstrated promising efficacy against GBM. However, like chemotherapeutic agents, systemically administered CAR-Ts suffer from limited access to the brain. We and others have demonstrated applications of focused ultrasound (FUS) with FDA approved microbubbles to amplify the deposition of ultrasound energy into tissue, inducing a variety of biological effects. Under certain ultrasound conditions, FUS can temporarily and reversibly alter the permeability of the BBB to deliver exogenous compounds into the brain, which has been demonstrated in preclinical models and early-stage clinical trials in humans. We have developed an ultrasound image-guided FUS therapeutic platform specific for rodents, capable of consistent and accurate targeting of BBB disruption and delivery of therapeutics to the brain, a necessary requirement for the proposed studies. Furthermore, we have identified the immunoregulatory protein B7-H3 as an attractive CAR-T target which is highly expressed in clinical GBM isolates but not in normal tissue. We have developed both murine B7-H3.CAR-Ts, which demonstrate efficacy in xenograft models, and human B7- H3.CAR-Ts, which are currently being evaluated in phase 1 clinical trials for safety and antitumor activity at our facility. Our team is therefore in a unique position to study the potential application of FUS to improve CAR-T therapy for GBM. Specifically, we hypothesize that FUS-mediated BBB disruption will result in increased trafficking of systemically administered B7-H3.CAR-Ts to GBM tumors; and FUS therapy will improve overall anti-tumor efficacy of B7-H3.CAR-Ts by modulating the immunosuppressive tumor microenvironment. These hypotheses will be addressed by 1) first establishing the effects of FUS with varying parameters to increase trafficking of B7-H3.CAR-Ts to GBM tumors, 2) determining B7-H3.CAR-T biodistribution, proliferation and persistence over time following systemic administration with FUS therapy, and finally 3) a pilot study to determine the impact of FUS therapy on the tumor immune microenvironment and how those changes augment B7- H3.CAR-T efficacy. Success of this biomedical engineering project would pave the way for further technology development as well as exploration and clinical translation of the synergy of these two relatively new and powerful technologies to make a substantial impact in the lives of GBM patients.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY Autistic females are diagnosed at a significantly lower rate and at later ages than males. As a result, much less is known about the female autism phenotype. Studies have indicated that diagnostic tools may be biased towards males and that findings characterizing the autism phenotype may fail to capture females due to little or no female representation in their sample. Emerging research has identified unique sex differences in the autistic population across several modalities, including language. The handful of studies that have characterized language use in autistic females have done so through examining word use, identifying nuanced ways that autistic females use discourse markers, pronouns, and emotional words. These differences may contribute to autistic females’ heightened social motivation, using language to compensate for social difficulties, although no study has directly explored this possibility. We propose to bridge this gap through a focus on speech prosody, an understudied area of pragmatic language that allows individuals to convey meaning and emotion through speech. Prosody plays an important role in social-communication and in the early detection of autism, yet few studies have included females in their sample and no study has closely examined potential sex differences. In Aim 1, we will characterize differences in key prosodic markers of pitch and tempo in young children (4-8 years) by sex, diagnosis, and conversational content using a novel adaptation of natural language sampling methodology. In Aim 2, we will use multi-modal data collected through an ongoing study (R01-HD103895-01A1) to explore the relationship between speech prosody, social traits, and formal language. We hypothesize that the autistic (ASD) group will significantly differ from the non-autistic (NA) group in multiple measures of pitch and tempo, and that these differences will be exacerbated during conversation around a “special interest” topic. We predict that while ASD females will display more “typical” prosody in common prosodic measurements of pitch, they will be differentiated from ASD males and NA females in more socially complex measurements of intonation, speech rate, and pause. We predict overall negative relationships between social measurements and speech prosody markers associated with ASD: These relationships will be stronger for ASD females than males, while relationships between speech prosody and language scores will be stronger for ASD males. Training aims include advancing Ms. Putnam’s skills in multi-modal methodologies, eye tracking, language processing, and statistical analyses. This project will enhance current understanding of speech prosody and how it relates to other important aspects of the autism phenotype. By further characterizing the female autism phenotype, this work will contribute to more timely diagnoses and comprehensive care of this understudied population.

NIH Research Projects · FY 2025 · 2023-12

PROJECT SUMMARY This proposed study will comprehensively stage individual disease burden and mortality risk at initial diagnosis and across the course of care for patients with chronic limb threatening ischemia (CLTI). This debilitating, progressive condition is the most severe form of peripheral artery disease, and successful treatment requires ongoing combinations of endovascular or open revascularization, medical and wound/podiatric care. Because CLTI manifests in the extremities, limb-based classification systems like the existing Wound, Ischemia, and foot Infection (WIfI) staging system are important tools; however, the goals for CLTI treatment are often broader than limb salvage and include freedom from recurrent disease and survival. Despite the availability of multiple life- and limb-preserving treatments, success in improving health outcomes of patients afflicted with CLTI has been limited. The recently completed BEST- CLI randomized controlled trial with 1830 adult participants determined that surgical vein bypass is the most effective initial revascularization approach for patients with CLTI, but almost 10% required re-intervention to maintain blood flow to the limb, 10% underwent major limb amputation, and 33% died within two years. The need for frequent re-intervention for patients in the BEST-CLI study demonstrates that long-term CLTI and survival outcomes are dependent on the sequence of treatments beyond the initial revascularization strategy. Yet, how post-primary revascularization treatment impacts outcomes among different types of patients is unknown. Our proposed study addresses these limitations. Using precision medicine analytics and machine learning, we will leverage the rich BEST-CLI trial data to identify clusters of limb characteristics, anatomic patterns of atherosclerosis, and comorbidities that associate with differing levels of CLTI-free survival. Moreover, with the BEST-CLI longitudinal patient assessments, we will quantify the impact of different post-primary revascularization treatments on CLTI-free survival. This study will provide a deeper understanding of the limb- and life- preserving effect of different combinations of medical and surgical treatments over time, especially among patients who do not respond to first-line surgical interventions. Together, our results will help clinicians set expectations and, if warranted, change practice based on likely CLTI-free survival at initial diagnosis and over the lifespan of each patient. In the future, the evidence generated from our study will be incorporated into precision medicine clinical trials such sequential multiple assignment randomized trials (SMARTs) for discovering and testing optimal, individually tailored treatment regimens for CLTI.