George Washington University

NIH Research Projects · FY 2026 · 2024-03

ABSTRACT Tumor antigen-triggered T cell receptor (TCR) signaling in the cytoplasm and transcriptional/epigenetic control in the nucleus are central to CD8+ T cell-dependent antitumor immunity. However, strong and persistent antigen stimulation leads to immune exhaustion. Compared to tumor drainage lymph nodes, the tumor microenvironment (TME) contains higher amounts of tumor antigens and their prolonged presentation to tumor- infiltrating CD8+ T cells can exacerbate T cell exhaustion and immune evasion. Except for co-inhibitory receptors, it is largely unknown how T cells adapt to the antigen-rich milieu of TME. Better understanding of the mechanisms underlying TME-unique regulation of T cell functions could inform novel therapies aimed at reinvigorating the antitumor potential of exhausted CD8+ T cells. The overarching scientific objective of our proposed work is to determine how negative elongation factor (NELF), known for its function in RNA polymerase II (Pol II) pausing, regulates CD8+ T cell activities in a TME- unique manner. Our preliminary work suggests that NELF is both necessary and rate-limiting in promoting the antitumor activity of CD8+ T cells. Of note, this NELF function is unique to CD8+ T cells within the TME, but not in tumor drainage lymph nodes. Furthermore, our published and unpublished data point to potential dual actions of distinct subcellular pools of NELF: while nuclear NELF participates in transcriptional and epigenetic programs, cytoplasmic NELF attenuates TCR signaling. We hypothesize that the combined actions of NELF likely contribute to stronger T cell memory and less T cell exhaustion. Built upon rigorous scientific premises and multidisciplinary expertise, we will validate this novel hypothesis by deciphering the TME-unique function of NELF (Aim 1), interrogating its dual molecular actions in the nucleus and cytoplasm (Aim 2), and exploring the utility of NELF overexpression in boosting efficacy of anticancer immunotherapies (Aim 3). The concept that NELF has a TME-unique function in antitumor immunity is novel. Furthermore, the proposed dual actions of NELF clearly deviate from the Pol II pausing-exclusive paradigm of NELF biology. The proposed work in this multi-PI R01 proposal promises to shed light on a previously unappreciated regulator of TME-unique functions of tumor-infiltrating T cells, with potential for a sustained impact on the understanding of antitumor immunity. From a translational perspective, discoveries from this project could inform more effective approaches to boost T cell memory and overcome T cell exhaustion, thus helping address a significant barrier and unmet clinical need in anticancer immunotherapies.

NIH Research Projects · FY 2026 · 2024-03

ABSTRACT Increasing evidence demonstrates that suboptimal conditions in the womb, including poor maternal nutrition, contributes to obesity and cardiometabolic disease risk. One potential contributor to poor maternal nutrition is chronic consumption of non-nutritive sweeteners (NNS). Studies indicate that approximately 30% of pregnant women and 44% of lactating women in the United States regularly consume NNS. However, a growing body of evidence demonstrates that NNS consumption is paradoxically associated with obesity and metabolic risk factors. While effects of NNS consumption during pregnancy and lactation on maternal and infant metabolic health are not well understood, the available evidence suggests cause for concern. For example, NNS consumption during pregnancy increases maternal blood sugar and induces fetal hypoglycemia and growth restriction in mice – both of which are risk factors for offspring obesity. Further, three prospective cohort studies in humans have reported an association between maternal NNS consumption and childhood weight or adiposity, although another found no relationship. Our own pilot data indicate that NNS consumption during pregnancy increases infant fat mass and disrupts the infant gut microbiome at 1 month of age. However, the limited, contradictory, and observational nature of the human evidence collected to date has made it difficult to establish guidelines around NNS consumption during pregnancy. We therefore propose the first randomized controlled trial (RCT) to examine the effects of NNS during pregnancy and lactation. We will recruit 324 pregnant women who frequently consume NNS (≥7 servings/week) and examine the effects of NNS restriction on maternal and infant metabolic outcomes. Pregnant women will be randomly assigned to one of three groups: 1) no NNS restriction (control); 2) NNS restriction during lactation only; and 3) NNS restriction during pregnancy and lactation. The intervention will consist of monthly consultations with a dietitian to provide counseling on replacement of NNS consumption with unsweetened food and beverage alternatives and home delivery of such alternatives. Adherence will be assessed via 24-hour dietary recalls and by objectively measuring NNS concentrations in urine and breast milk. Our primary aims will be to determine the effects of NNS-restriction on: 1) infant body composition at 1 and 6 months of age; 2) maternal glucose tolerance at 24-28 weeks’ gestation; and, 3) infant gut microbiome and fecal metabolites at 1 and 6 months of age. We hypothesize that NNS restriction among pregnant women who habitually consume NNS during pregnancy and/or lactation will reduce infant fat mass (compared to a no NNS restriction (usual NNS intake) control group), via improvements to maternal glycemia and the infant gut microbiome and metabolome, with the greatest benefit in the pregnancy and lactation group. This research could shape recommendations around NNS consumption during pregnancy and lactation, thereby potentially reducing the global burden of obesity and cardiometabolic disease.

NIH Research Projects · FY 2025 · 2023-09

incidence and longer persistence of high-risk PROJECT SUMMARY/ABSTRACT (WLH) bear a disproportionate risk of invasive cervical cancer (CC) due to greater human papillomavirus (HPV) infection. Despite the availability of effective Women living with diagnosed HIV CC screening methods, there is low CC screening uptake among WLH. Reasons for suboptimal CC screening uptake among WLH are complex and multifactorial. Multilevel interventions that simultaneously address modifiable barriers to screening at different levels of influences are therefore needed to effectively reduce CC disparities among WLH. To achieve the World Health Organization’s recommended global target to eliminate CC by 2030, accelerated progress in complex health problems may also likely come from the integration of social innovations and technological advances. The overall objective of this proposed K01 project is to conduct formative research and pilot test the provider-level and patient-level components of the My Self-Sampling for HPV Awareness, Results, and Empowerment (MySHARE+) intervention. MySHARE+ aims to harness the power of technology and apply a multilevel approach to promote the adoption of CC screening (HPV self-sampling; Pap triage adherence) among under/never-screened WLH, a vulnerable population. The specific aims are to: 1) identify facilitators and barriers to implementing a healthcare provider prompt in a primary care setting and 2) conduct a pilot RCT to examine the feasibility, acceptability, and preliminary efficacy of a mHealth educational intervention in promoting CC awareness and HPV self-sampling among WLH. These aims align with the NCI’s mission to advance scientific knowledge to reduce the unequal burden of cancer and purposely addresses their areas of research emphasis in biobehavioral research and in improving health disparities by targeting multiple levels of influences. Study findings will generate implementation feasibility and acceptability data from the provider and patient perspectives, and inform a subsequent R01 that will test the impact of the MySHARE+ intervention in a large cluster randomized trial where the independent and overlapping effects of the different components can be evaluated. Strongly supported by mentors and advisors who are leading experts in the methodological and content areas specific to her long-term goals, Dr. Le’s proposed K01 includes a comprehensive training plan that will help her acquire new knowledge and advanced skills in: 1) quasi-experimental studies and cluster-RCTs using multilevel approaches, 2) digital health/technology-based interventions to promote patient-centered care, and 3) implementation science methodologies and frameworks. Over the 5-year award period, she will participate in mentored research, one-on-one and advisory/team meetings, didactic instruction, directed readings, training seminars, and scientific meetings. The aforesaid research and training activities will undoubtedly help Dr. Le achieve her overall academic career objective of leveraging academic- community-government partnerships to successfully execute and sustain community-driven and clinically-embedded intervention research supported by health information technology and the remote delivery of primary health care services to enhance patient-centered care. GW is well poised to support Dr. Le in undertaking these research endeavors, especially due to the expertise on faculty, its location and reputation in the city, and the institution’s high quality research setting.

NIH Research Projects · FY 2024 · 2023-09

Hookworms are serious parasites of humans and animals, causing anemia and even death in heavy infections and vulnerable populations. Nearly 500 million people are infected with hookworms, primarily in developing countries lacking sanitation infrastructure. Canine hookworms are the most important gastrointestinal parasite of dogs. Hookworms can be controlled using several anthelmintics, which in the absence of a vaccine, remain the only available control for hookworm infections. However, heavy use of anthelmintics has led to the development of genetic resistance to all the major anthelmintics used to treat parasitic nematodes in livestock. Increased use of anthelmintics to treat humans is predicted to lead to anthelmintic resistance (AR) in hookworms as well. Indeed, heavy use of anthelmintics in racing greyhounds has led to the emergence of multidrug resistant (MDR) canine hookworms (Ancylostoma caninum). These MDR isolates are resistant to all classes of anthelmintics approved for use in dogs in the US. Recently, MDR hookworms have spread from greyhounds into the general pet dog population and are predicted to continue this spread in the future. Not only will MDR hookworms negatively affect the ability to treated infected dogs, they will also severely limit treatment options for zoonotic hookworm infections in humans such as cutaneous larva migrans, eosinophilic enteritis and diffuse unilateral subacute neuroretinitis. Therefore, there is an urgent need to develop new drugs to treat infections with MDR hookworms. Development of anthelmintics requires adult parasites to test potential compounds for activity in vitro, and infected hosts to test the efficacy of potential drugs. This is hampered by the lack of a small animal model for hookworm infections in which to test new drugs. Ancylostoma caninum can currently only be maintained in dogs, making anthelmintic development and testing costly and ethically problematic. Therefore, we propose a two-pronged approach to developing a rodent model for canine hookworm infection. In Aim 1, we will use drugs to immunosuppress potential rodent hosts before and during infection to determine if the animals can be permissive to MDR hookworm infection. We will also test immunodeficient rodents for their ability to host A. caninum. In Aim 2, we will use the rodent model to test current anthelmintics as a proof of principle. A small animal model of canine hookworm infection would greatly facilitate the development of anthelmintics effective against MDR worms and would provide insights into the development of AR in hookworms that are applicable to hookworm infections of humans.

NIH Research Projects · FY 2025 · 2023-09

Project Summary The goal of this R01 proposal is to develop an ultra-soft, fully implantable, wireless label-free cardiac mapping and modulation system and apply it to identify chronic electrophysiological and metabolic changes and their links during heart failure (HF) development, progression, and pacing treatment in unrestrained conscious animals at cellular and whole heart levels. To achieve this, a miniaturized, mechanically compliant platform that integrates high-density, high-resolution sensing and modulation channels with wireless energy harvesting, storage, control, and data communication module is proposed. The resulting systems will greatly reduce motion artifacts and allow bidirectional high-content electrical and metabolic mapping and pacing in live animals. Those devices are innovative because they directly address the current limitations in chronically quantifying the individual roles and interplay between vital cardiac biophysical parameters during heart disease pathogenesis and will be used to fundamentally investigate the complex disease mechanisms involved in pathophysiological conditions leading to lethal HF and its therapeutic treatment. Once realized, this technology will be highly valuable to the cardiac research community. In the long term this work will enable closed-loop multiparametric cardiac mapping and pacing systems and offer new approaches to study the precise mechanisms and optimize the diagnostic and therapeutic strategies of other life-threatening heart diseases beyond HF. The three specific aims are: Aim 1 will establish ultra-soft multimodal cardiac systems for label-free cellular-resolution mapping of the excitation-contraction-metabolic waves and cardiac pacing. The mechanically compliant highly stretchable systems consist of high-density arrays (~300 channels in total) of (1) transparent microelectrodes for electrical mapping and stimulation; (2) multicolor micro-light-emitting diodes, and micro-photodetectors to excite and measure the autofluorescence of major endogenous fluorescent markers of cellular energy metabolism. Aim 2 will develop fully implantable wireless schemes for power harvesting, storage, control, and data communication to chronically operate the platforms in Aim 1 within a closed thoracic cavity in freely behaving small animals, which is beyond any possibility supported by current techniques. Graphical user interfaces will be developed for device configuration, real-time bidirectional control, data acquisition and processing. The integrated systems will be characterized, validated, and optimized by iterative benchtop measurements. Aim 3 will systematically investigate the precise mechanisms of HF pathogenesis and therapy using a battery of tests in rat models of HF. The functions of the proposed systems will be assessed in both ex vivo and in vivo studies. The implantable cardiac devices will identify the individual roles and links between local metabolic and electrical properties during different time points and stages of HF development and progression, and thoroughly evaluate the effects of cardiac-resynchronization therapy.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT There is a high global burden of untreated postpartum depression. Untreated postpartum depression presents acute and long-term risks to a mother’s mental and physical health, as well as her child’s survival, health, and development. Although there is increasing willingness among funders and health systems to involve nonspecialists, such as community health workers, in the delivery of psychological interventions for postpartum depression, the outcomes of these non-specialist interventions have been mixed. Digital technology, specifically the use of passive sensing data collection to learn more about a mother’s life and support behavior change, has the potential to improve the effectiveness of psychological interventions delivered by non-specialists. For the proposed R21/R33, we will build on preliminary development work of Sensing Technologies for Maternal Depression Treatment (StandStrong) to demonstrate feasibility, acceptability, and functionality for postpartum depression treatment. StandStrong is a digital platform consisting of two mobile apps, internet-of-things (IoT) passive sensors, and a data analysis engine built using modern machine learning approaches. StandStrong incorporates passive sensing data collection from mothers and their infants that can be monitored by nonspecialist counselors and then incorporated to better personalize psychological treatment. For Aim 1 (R21 phase), we will use human-centered design to iteratively refine the StandStrong platform and associated implementation material. For Aim 2 (R21 phase), we will evaluate the StandStrong platform and associated contents according to acceptability, feasibility, usage, benefit, and validity. If milestones are met for the R21, we will proceed to the R33 stage. In the R33 stage, for Aim 3 we will conduct a pilot randomized controlled trial (RCT) comparing a psychological treatment as usual with the psychological treatment supplemented by the StandStrong passive sensing platform with 112 depressed mothers (56 in each arm). For Aim 4 of the R33 phase, we will identify parameters for sustainability and scalability, as well as costing the intervention, to ultimately prepare for a fully-powered RCT in the United States with future R01 funding. Successful completion contributes to the NIMH’s 2020 Strategic Plan to tailor existing interventions and develop innovative service deliver for mental health interventions. This project will contribute to NIMH’s Opportunities and Challenges of Developing Information Technologies through improving treatment of mental health conditions through use of passive sensing technologies.

NIH Research Projects · FY 2024 · 2023-08

Abstract Cardiovascular diseases (CVD) are the primary cause of premature death worldwide, and risk factors such as insulin resistance and type 2 diabetes are increasing in prevalence. Therefore, more effective preventive interventions to improve cardiometabolic health are urgently needed. Circulating branched-chain amino acids (BCAA) are strong predictors of type 2 diabetes and CVD. Furthermore, experimental and genetic studies indicate that BCAA overload has adverse cardiometabolic effects. Thus, BCAA levels may be novel targets for preventive interventions. In addition, BCAA levels of individuals may inform personalized interventions. Evidence is accumulating that diet composition, physical activity, and weight loss can reduce plasma BCAA levels by affecting the dietary supply, gut microbiota production, or catabolism of BCAA. However, evidence on the impact of lifestyle factors on BCAA levels is mainly based on small experimental studies and cross- sectional studies. Larger trials and longitudinal studies are crucial to determine whether lifestyle exposures can substantially affect BCAA levels. Furthermore, research on the health consequences of BCAA levels has focused chiefly on the impact of BCAA levels on glucose metabolism, but adverse effects on other CVD risk factors such as inflammation, blood pressure, and dyslipidemia are plausible. The PREMIER study is a randomized trial of the impact of behavioral lifestyle interventions on CVD risk factors with detailed information on diet and physical activity based on interviews and biological measurements. We will measure serum BCAA and other metabolites using targeted quantitative NMR metabolomics in serum samples of 729 participants at baseline and after six months of lifestyle interventions. Our specific aims are as follows: 1). To assess the impact of the randomized lifestyle interventions on circulating BCAA levels over six months. We will evaluate the effect of the ‘established lifestyle intervention,’ including weight loss and increases in physical activity, and the effect of the intervention with added counseling to improve dietary patterns as compared with the control arm. 2). To evaluate specific dietary factors, physical activity, and changes in adiposity in relation to BCAA changes. We will examine changes in physical activity, fitness, protein and BCAA intakes, and healthy plant-based dietary patterns. 3). To evaluate whether changes in BCAA levels affect cardio-metabolic risk factors, including blood lipid profiles, blood pressure, and markers of inflammation and insulin sensitivity. We will also estimate the proportion of the effects of lifestyle changes on cardiovascular risk factors explained by reductions in BCAA levels. Our findings may inform dietary and lifestyle recommendations that consider the impact on BCAA levels.

NIH Research Projects · FY 2024 · 2023-08

Project Summary/Abstract Advances in B cell biology and molecular virology have enabled the discovery, characterization, and commercial development of broadly neutralizing antibodies (bNAbs). They are a promising immunotherapy that can be incorporated in many strategies for the treatment and/or cure of HIV-1. Antibodies have many effector functions beyond neutralization and their most important mechanism of action for treatment/cure may be their ability to opsonize infected cells, tagging them for antibody-dependent cellular cytotoxicity or phagocytosis (ADCC/ADCP) by immune effector cells. It is not clear that bNAbs are mediating clearance of infected cells in clinical trials when they are passively infused into chronically infected people living with HIV (PLWH). An obvious reason for this lack of efficacy of bNAbs may be a lack of Env protein expression while on suppressive ART. Our lack of knowledge about the level of Env expression during HIV latency and latency reversal prevents effective use of bNAbs as therapeutics. To address this gap, the ability to detect low level Env protein expression in cells infected with different subtypes of HIV-1 is needed. The Bosque lab has published an ultrasensitive method to detect p24 Gag protein down to the fg/ml level and has preliminary data for a newly developed Env assay. Here we propose to use these assays to detect low levels of Env and Gag protein in a well-described model of latency and compare the data to Env surface expression as measured by flow cytometry (Aim 1). Within these assays, we have the flexibility to test Env detection using bNAbs targeting different epitopes to probe for Env conformation (i.e., trimer, monomer, etc) and extend these analyses to diverse HIV isolates from different subtypes to measure differences in Env expression and latency attributable to genetically diverse virus (Aim 2). Knowing these crucial factors about Env and Gag expression in a carefully controlled model of latency will allow us to further investigate HIV protein translation in future studies using cohorts of PLWH. These studies will improve the ability of therapeutics using Env-targeting strategies to target latently infected cells.

NIH Research Projects · FY 2025 · 2023-07

ABSTRACT Women with germline mutations of BRCA1 have about an 80% lifetime risk of developing breast cancer. All somatic cells in a germline BRCA1 mutation carrier harbor the same mutant copy of the BRCA1 gene, yet BRCA1 research is almost exclusively focused on BRCA1’s role in repair of double-strand DNA breaks in the breast epithelium. This is due to the cardinal tenet of the current BRCA1 paradigm — increased genetic instability of the breast epithelium solely drives development of BRCA1 mutation-related breast cancer. In addition, current prophylactic risk-reducing surgeries are aimed at curtailing tumorigenesis of breast epithelial cells of BRCA1 mutation carriers. In contrast, virtually nothing is known about whether BRCA1 heterozygosity in cells beyond the breast epithelium could also contribute to BRCA1-related tumorigenesis. Neoplasia is antagonized by the ability of the immune system to detect and eliminate neoplastic cells. Clearly, impaired antitumor immunity enables immune evasion and tumor outgrowth. We recently found that women with BRCA1 germline mutations have less abundant circulating CD8+ T cells versus age-matched donors with wild- type BRCA1 genes. In support of this clinical finding, heterozygous knockout of Brca1 specifically in CD8+ T cells of mice is sufficient to attenuate antitumor immunity. Based on our preliminary data, we hypothesize that T cell- intrinsic function of BRCA1 in enhancing antitumor immunity contributes to the overall tumor suppressor activity of BRCA1. We further propose that in BRCA1 mutation carriers, increased genomic instability in the breast epithelium and attenuated antitumor immunity in CD8+ T cells act together to elevate cancer incidence. To validate this novel hypothesis, we will combine our established team’s tools and expertise in cancer biology, tumor immunology, and transcriptional regulation through the following three Specific Aims: (1) Discern the impact of T cell-intrinsic BRCA1 on antitumor immunity and immunotherapy, (2) Elucidate the molecular basis for BRCA1’s function in antitumor immunity, and (3) Examine BRCA1 mutation-related T cell deficiency in human samples. Our proposed work represents a new direction in elucidating a previously unappreciated tumor-suppressing function of BRCA1 in immune cells. The concept to be validated clearly departs from the prevailing paradigm regarding BRCA1-related cancer etiology, which is focused on the breast epithelium and DNA repair. From a translational perspective, our proposed study focusing on BRCA1 in immune cells will likely inform development of novel immune-boosting strategies for women with BRCA1 germline mutations. Thus, this line of work has potential for far-reaching and sustained impact on breast cancer cancer risk reduction.

NIH Research Projects · FY 2026 · 2023-07

ABSTRACT American Indian/Alaska Native (AI/AN) populations experience some of the most extensive health disparities in the U.S. for any demographic group. Increasing evidence shows that indigenous historical trauma (IHT) is a likely causal factor for AI/AN health disparities. However, recent research has only begun the process of defining and operationalizing IHT as a construct that could be measured and assessed as a health determinant, and the instruments now available focus primarily on historical loss experience and associated symptoms, though the experience and effects of IHT are likely to be more extensive. Progress is needed to develop validated assessment tools that capture broader and cross-generational aspects of the phenomenon. To address this specific gap in the literature and contribute to the reduction of health disparities among AI/ANs, we propose community-based research that brings together complementary strengths of the Department of Prevention and Community Health (DPCH) at the George Washington University Milken Institute School of Public Health (GWSPH), the Eastern Band of Cherokee Indians (EBCI), and Western Carolina University (WCU) in a partnership to: 1) Use systematic qualitative methods, cultural models theory, and a community participatory approach to elicit/validate characteristics and manifestations of an expanded IHT construct (to include those identified in previous research), followed by scale development procedures to develop and pre- test an expanded IHT instrument that integrates these broader factors; and 2) administer the newly developed IHT scale and health assessment instrument to a sample of 400 Tribal participants to test its internal consistency, reliability, and factor structure. We will additionally evaluate the associations between IHT and behavioral risks/outcomes and priority health conditions prioritized by EBCI, including substance abuse and related issues, violence and abuse, diabetes, stress and depression. Results will be disseminated to EBCI and to the field.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY A key Healthy People 2030 Tobacco-Use Objective is to eliminate state preemption of local tobacco control. However, little published research has rigorously quantified the impacts of state preemption of local tobacco control, particularly: 1) across states and over time; 2) across a broad range of tobacco control efforts (e.g., restrictions on advertising, youth access); 3) across a range of key outcomes (e.g., youth and adult tobacco use, secondhand smoke exposure [SHSe], tobacco retail); and 4) with regard to potential disproportionate impacts among certain subgroups that face tobacco-related disparities (e.g., racial/ethnic minorities, lower socioeconomic status [SES], those in rural settings). As initial evidence providing a basis for this proposal, our RWJF-funded research concluded that state smoke-free air preemption was associated with an increase in adult smoking prevalence and affect counties differently. This proposal will build on this work and advance the literature by filling the aforementioned gaps in the existing research. The long-term goal of this research is to inform policies to reduce tobacco use and related health disparities, as well as to inform policy efforts more broadly, particularly as public health issues, like firearm safety, nutrition policies, and COVID-19 mitigation measures, are increasingly subject to state preemption. Our objective is to advance our understanding of whether—and to what extent—enactment or repeal of state preemption on local tobacco control is associated with disparities in tobacco use, SHSe, and retail (sales, retailer density). Our central hypothesis is that state preemption results in disproportionately high tobacco use, SHSe, and retail in communities with greater racial/ethnic minorities, of lower SES, and/or in rural settings. Our rationale is that, by preempting local tobacco control, state governments deprive localities of a crucial tool for reducing the burden of tobacco in their communities, potentially widening disparities. We will analyze national data from 1999 to 2021 to examine the impacts of enactment or repeal of state preemption of 5 state tobacco preemption (i.e., smoke-free policy, advertising, licensure, youth access, and taxation). Our specific aims are to examine state preemption on tobacco control in relation to changes in: 1) individual-level tobacco use (including e-cigs) in adolescents and adults and nonsmokers' SHSe, and potential disproportionate impacts among certain subgroups of individuals; and 2) sales of tobacco products and tobacco retail density over time, and potential disproportionate impacts among communities representing those facing tobacco-related disparities. We will employ multilevel models and crosswalk/merge (state/local laws, outcomes, sociodemographics) data from various sources at different geographic levels. This study will have high impact, as it will: 1) add to the evidence base regarding state preemption impact on population outcomes and mechanisms of impact; 2) inform efforts to reduce tobacco use and related disparities; and 3) engage key tobacco control and policy experts in research dissemination.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY Broadly neutralizing antibodies (bNAbs) are a promising immunotherapy that can be incorporated in many strategies for the treatment and/or cure of HIV-1. There is a clear association between the neutralization sensitivity of virus and how effective bNAbs are in suppressing virus replication during clinical trials, but the precise nature of this relationship is still not clear. Individual clinical trials have reached different conclusions about the utility of pre-screening participants’ virus for in vitro neutralization sensitivity before enrollment into clinical trials mainly due to two obstacles: length of assay time and difficulty in obtaining the correct samples to test. The Bosque lab has recently published an ultrasensitive method to detect p24 gag protein down to the fg/ml level, and the assay was validated in ex vivo cells from PLWH. Here we will apply this novel methodology to address these 2 obstacles by developing an assay that gives an indirect readout of neutralization sensitivity but in a very short amount of time and directly in cell lysates. We will achieve this ultra-sensitive Env binding assay by developing it in three parts for the first R61 phase: Aim 1 will optimize the capture and detector antibodies by testing a matrix of bNAbs against diverse pseudoviruses with known neutralization sensitivities, Aim 2 will optimize biological matrices by testing Env detection in plasma and cell lysates, and Aim 3 will determine assay precision for ratios of sensitive and resistant virus in a diverse viral quasispecies. We will validate and qualify this ultra-sensitive Env binding assay during the second R33 phase in two parts: Aim 4 will validate the assay through post-hoc testing of well-studied clinical trial samples. Aim 5 will implement correct quality systems for this assay to prepare for CLIA qualification. Our multi-disciplinary team has all the expertise necessary to achieve these aims that, when completed, result in an ultra-sensitive binding assay for Env protein to screen clinical trial participants that is time-efficient, accurate and qualified.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Broadly neutralizing antibodies (bNAbs) are a promising immunotherapy that can be incorporated in many strategies for the treatment and/or cure of HIV-1. There is a clear association between the neutralization sensitivity of virus and how effective bNAbs are in suppressing virus replication during clinical trials, but the precise nature of this relationship is still not clear. Individual clinical trials have reached different conclusions about the utility of pre-screening participants’ virus for in vitro neutralization sensitivity before enrollment into clinical trials mainly due to two obstacles: length of assay time and difficulty in obtaining the correct samples to test. The Bosque lab has recently published an ultrasensitive method to detect p24 gag protein down to the fg/ml level, and the assay was validated in ex vivo cells from PLWH. Here we will apply this novel methodology to address these 2 obstacles by developing an assay that gives an indirect readout of neutralization sensitivity but in a very short amount of time and directly in cell lysates. We will achieve this ultra-sensitive Env binding assay by developing it in three parts for the first R61 phase: Aim 1 will optimize the capture and detector antibodies by testing a matrix of bNAbs against diverse pseudoviruses with known neutralization sensitivities, Aim 2 will optimize biological matrices by testing Env detection in plasma and cell lysates, and Aim 3 will determine assay precision for ratios of sensitive and resistant virus in a diverse viral quasispecies. We will validate and qualify this ultra-sensitive Env binding assay during the second R33 phase in two parts: Aim 4 will validate the assay through post-hoc testing of well-studied clinical trial samples. Aim 5 will implement correct quality systems for this assay to prepare for CLIA qualification. Our multi-disciplinary team has all the expertise necessary to achieve these aims that, when completed, result in an ultra-sensitive binding assay for Env protein to screen clinical trial participants that is time-efficient, accurate and qualified.

NIH Research Projects · FY 2026 · 2023-04

Non-communicable diseases (NCDs) account for >70% of deaths each year globally and approximately 86% of deaths in the US. NCDs have a particularly profound impact in low- and middle-income countries (LMICs), where about 73% of NCD-related deaths worldwide occur. Critical to decreasing the NCD burden is reducing modifiable NCD risk factors (e.g., tobacco/alcohol use, poor nutrition), which also disproportionately impact LMICs. Importantly, addressing social determinants (SDs) of health is critical in reducing the global NCD burden. SDs – or conditions in which people are born, live, learn, work, play, and age – are important drivers of NCDs, and impact both modifiable risk factors (e.g., smoking) and disease outcomes. SDs represent various domains including: healthcare access/quality, social/community context, neighborhood/built environment, economic stability, and education access/quality. Thus, key approaches to NCD prevention and control include addressing SDs of NCD risk factors via multilevel interventions. Notably, disadvantages related to SDs are especially prominent in LMICs. The ACCESS program (i.e., the Armenia [AM]-US Collaboration to Address Chronic Disease via Education in Social Determinants Science program) will be the first program of its kind and will provide a model for studying and addressing pressing NCD-related health priorities among high-risk populations and in low-resource settings. AM is an LMIC that faces particular NCD-related challenges, as NCDs account for 93% of all deaths and the top 10 causes of death, premature death, and disability in AM. Relatedly, AM is plagued with profound NCD risk factors, as AM has the 11th highest male smoking rate globally (52%), ranks in the top 40% of countries in alcoholism rates, and has a 48% prevalence of overweight/obesity. Moreover, understanding the SDs of NCD and related risk factors in high-risk populations and in low-resource settings, like AM, may provide unique scientific insights that translate to such settings and populations in the US and globally. ACCESS is a strategic collaboration between George Washington University (GW), American University of Armenia (AUA), AM’s Ministry of Health (MOH), and its entities (e.g., NIH) – thus capitalizing on the unique scientific talent in our partnering Armenian organizations and their commitment to developing an effective, sustainable training program to enhance SDs and NCD research capacity, emphasizing high-risk populations and delivery in low-resource settings, including those in the US and more broadly. Aim 1 will establish a formal SDs and NCD prevention and control research training program and related institutional infrastructure for MPH training among doctoral-level investigators that contributes to US-based training and infrastructure. Aim 2 will advance global NCD research priorities via mentored research focused on SDs of NCD risk factors among future NCD research leaders. Aim 3 will catalyze NCD-related research dissemination and knowledge translation to inform high-impact interventions. ACCESS will establish a bidirectional exchange of data and expertise to identify and address key SDs of NCDs, informing and supporting the development, testing, and dissemination of evidence-based interventions that improve and protect health in the United States and worldwide.

NIH Research Projects · FY 2026 · 2023-04

(<30 lines) Youth-onset type 2 diabetes (YO-T2D) is increasingly prevalent in parallel with the obesity epidemic, yet effective treatment and prevention strategies are limited. The physiologic increase in insulin resistance occurring during puberty, in combination with obesity-related insulin resistance, enhances the risk of T2D. Yet, it remains unclear why some youth progress through puberty with intact β-cell function, while others do not, despite similar phenotypic and metabolic characteristics. More information is needed regarding the unique events during puberty to better understand 1) the basic pathophysiology of glucose control, insulin sensitivity, β-cell function, and T2D risk in youth, 2) differences among girls and boys, populations at highest risk, and urban and rural geographies, and 3) the potential contribution of other risk factors including psychological, behavioral, and social and external contexts. Importantly, this research needs to address the timeline of pathophysiology and progression from normoglycemia or prediabetes to YO-T2D. The DISCOVERY of Risk Factors for Type 2 Diabetes in Youth (DISCOVERY) study provides a unique opportunity to characterize the risk progression profile and mechanisms underlying the development of YO-T2D, and evaluate the effects of modifiable and non-modifiable risk factors. Ultimately, the results of this study will establish a basic pathophysiology to inform future studies aimed at achieving target glycemia, improving insulin sensitivity, preserving β-cell function, and/or preventing YO-T2D. To address this goal, DISCOVERY will recruit, enroll, and follow a nationally-representative cohort of 3,600 at-risk obese youth in early puberty; extensively phenotype them as they transition through puberty; and characterize the course of decline and dysfunction in pathophysiological indicators that lead to YO-T2D. The expected duration of the DISCOVERY is 5 years, including planning, recruitment, follow-up, analysis, and reporting. In addition, DISCOVERY will store longitudinal biospecimens and genetic material with the intention of acquiring additional ancillary funding to pursue analysis of emerging indicators. The Biostatistics Research Center (BRC) will enhance the value of the DISCOVERY study by 1) overseeing all operational aspects of DISCOVERY, 2) providing administrative resources and logistical support for DISCOVERY, and 3) providing scientific and biostatistical expertise for DISCOVERY. Through effective organization, communication, and support, and by promoting a collaborative environment, the BRC will provide the framework and infrastructure for the Consortium to successfully recruit a cohort of early pubertal youth at-risk for developing prediabetes and T2D, deeply phenotype them through puberty, and ultimately contribute to a better understanding of the pathophysiology of YO-T2D.

NIH Research Projects · FY 2025 · 2023-04

Project Summary The proportion of Americans over the age of 65 is expected to increase over the next few decades and the number of those afflicted with Alzheimer’s disease and related dementias (ADRD) is also expected to increase substantially. To adequately serve this growing population, we need to make considerable progress in understanding ADRD and in developing evidence-based practices for prevention and care. As the US population ages, it is also becoming more racially and ethnically diverse. Despite this fact, the racial and ethnic diversity of those focusing on ADRD research does not reflect the diversity of the population that bears the high burden of ADRD. This is problematic, as the ability to be responsive to ADRD needs of such a population will require culturally informed approaches and diverse investigators with appropriate knowledge and skills. Information about and opportunities to enter the ADRD research workforce are not equally available to all, which may be why few students from racial and ethnic minority backgrounds pursue graduate study or careers in this area. In order to address these issues, we propose to establish a summer program and mentorship network that expands the pipeline of students from underrepresented groups who decide to pursue graduate study and, ultimately, careers in ADRD-related research areas. The proposed program will be jointly administered by two world-class research institutions in two of the nation’s biomedical and public health research hubs: George Washington University (GWU) in Washington, DC and Boston University (BU) in Boston, MA. Faculty from GWU’s Institute for Brain Health and Dementia will work in collaboration with faculty from BU’s Framingham Heart Study Brain Aging Program (FHSBAP) to mentor program participants as they work on ADRD-related research projects. The program will provide structured and suitably-tailored didactic, research, and social experiences for participants that will serve to elicit and magnify interest in ADRD research, develop skills that will make them competitive applicants for graduate study, and provide a network of mentors who will engage with them as they navigate the path from undergraduate to graduate study.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY/ABSTRACT Clamping and cutting the umbilical cord is the most common intervention in humans, occurring in 140 million annual births. To maximize expediency (manage 3rd stage of labor, initiate neonatal resuscitation), early cord clamping (ECC) is performed within ~30 sec of delivery. Recently, evidence has emerged on the health benefits of delayed cord clamping (DCC, waiting ~1-2 min before clamping). Among term neonates, multiple randomized controlled trials (RCTs) have shown that DCC transfers blood from placentas to newborns, resulting in less iron- deficiency anemia and improved neurodevelopmental (neuromotor) outcomes through 4 years, than with ECC. Despite advantages for healthy newborns, in view of their exclusion from previous RCTs, the best approach to cord clamping in higher-risk pregnancies, notably those complicated by a fetal diagnosis of critical congenital heart disease (CCHD), remains uncertain. Although one might assume that the benefits of DCC in low-risk new- born populations would translate simply to CCHD neonates, unique anatomic and physiologic differences in neonates with CCHD suggest that risks of DCC in this subgroup may differ from risks in neonates without CCHD. Thus, a treatment dilemma exists on the optimal cord clamping practice at birth among CCHD neonates. The proposed study, entitled CORD-CHD (Clamp OR Delay among neonates with Congenital Heart Disease) trial will be the first RCT to determine the effectiveness of DCC vs. ECC on postnatal and neurodevelopmental outcomes of CCHD neonates. Our preliminary data demonstrate that, among CCHD neonates, DCC results in lower global rank scores (GRS), indicative of better health outcomes, than ECC. GRS is a validated composite measure, based upon the worst outcome post-cardiac surgery or catheterization and reflects the need for com- plex intensive care. Given the absence of high-quality data, maternal outcomes will also be determined. We will leverage a network of sites with requisite infrastructures, established guidelines with high adherence rates and treatment fidelity, and track records of collaboration. Aim 1 is to test the hypothesis that, among CCHD neonates, DCC results in lower GRS (better outcomes) post-cardiac surgery or catheterization, based on a higher win-odds (win-ratio adapted to include ties), than with ECC. Aim 2 is to test the hypothesis that, among neonates with CCHD, DCC will result in better neuromotor outcomes at 22-26mos postnatal, based on a joint test of a bivariate outcome (Developmental Assessment of Young Children Second Edition motor score and the Hammersmith Neonate Neurological Exam) than with ECC. As a secondary objective, we will test the hypothesis that among CCHD neonates improved neuromotor profiles (General Movement Assessment) at 3-4mos mediate improved neuromotor outcomes at 22-26mos. Aim 3 is to precisely estimate the difference in the risk of maternal postpar- tum hemorrhage between DCC and ECC to evaluate safety among mothers who give birth to CCHD neonates. This trial will advance the care of CCHD neonates and provide the evidence called for by national and interna- tional organizations, creating a global impact on umbilical cord management among a vulnerable population.

NIH Research Projects · FY 2026 · 2023-03

Project Summary Cyclin-dependent kinase CDK4/6 inhibitor (CDK4/6i) in combination with aromatase inhibitors is the first-line treatment for ER+ advanced or metastatic breast cancer. Despite CDK4/6 inhibitors significantly improve overall survival of such patients, not all patients respond to these drugs and most patients whose tumors initially respond to CDK4/6i eventually develop acquired resistance. Although many efforts have been invested into the studying mechanism of resistance, CDK4/6i resistance remains a big challenge for HR+ breast cancer. Thus, it is urgent to develop new approaches to overcome resistance in CDK4/6i resistant breast cancer (CRBC). O-linked-N-acetylglucosaminylation (O-GlcNAcylation) is one type of glycosylation that occurs when a monosaccharide, O-GlcNAc, is added onto serine or threonine residues of proteins by O-GlcNAc transferase (OGT). O-GlcNAcylation is involved in a range of cellular activities and aberrant O-GlcNAcylation has been implicated in a host of diseases including cancer. However, the role of O-GlcNAcylation in cancer drug resistance remains largely unknown. Through an innovative quantitative high throughput combination screen (qHTCS) and follow-up extensive preliminary studies using cellular and molecular approaches, we identified a novel OGT-mediated mechanism regulating the resistance to CDK4/6i in ER+ breast cancer. The major objective of this proposal is to determine the role of OGT-mediated pathway in the regulation of CDK4/6i resistance in CRBC cells. Specifically, we will (1) investigate the molecular mechanism of how OGT-mediated pathway regulates CDK4/6i resistance in CRBC cells, (2) evaluate the effects of newly identified drug combinational treatments on palbociclib resistance using resistant PDX and syngeneic models, (3) conduct clinical study to further evaluate the correlation between OGT-mediated pathway and CDK4/6i resistance in tumors from patients. The completion of proposed studies will not only elucidate a novel mechanism regulating CDK4/6i resistance in ER+ breast cancer, but also provide an innovative therapeutic strategy to treat CRBC patients.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY The nasal cavity has a unique microbiome, which is expected to significantly affect local immune response and in turn, susceptibility of the host to respiratory viruses. However, we know little about these potential host- microbe interactions. Our long-term goal is to elucidate how microbiome-immune interactions and dynamics within the nasal cavity affect host susceptibility to infectious and inflammatory conditions. This project’s objective is to determine natural dynamics of correlations between nasal microbiome and local immune environment and how this relationship affects host susceptibility to respiratory viral infections. Our central hypothesis is that during homeostasis, the nasal microbiome impacts local antiviral immune defenses, including epithelial integrity, mucus properties, and antiviral cytokines and specific nasal microbiome-immune environments defined by low-inflammation and high-interferon levels will confer greater natural protection against respiratory viral infections. The rationale for this project is that our knowledge regarding the adult nasal microbiome and immune environment during homeostasis is limited. Addressing this gap can immediately improve our fundamental understanding of the adult nasal cavity and inform strategies that promote naturally occurring protection against respiratory viral infections. Aim 1. Elucidate dynamics of the homeostatic nasal microbiome and immune environment in adults. We will achieve this aim by studying (i) the homeostatic nasal microbiome and innate immune profile in a cross-section of adults (n = 400) and (ii) assessing short- and long-term dynamics of adult nasal environment by selecting and following 118 adults through monthly and quarterly dense sampling over a 12-month period. Aim 2. Determine how the nasal microbiome-immune environment affects susceptibility to an intra- nasal virus challenge. We will achieve this aim by studying how pre-vaccination nasal microbiome-immune environments impact post-vaccination viral infection and replication among adult FluMist® recipients (n = 200) per year in Years 1 and 2. Aim 3. Elucidate interactions between nasal microbiome, nasal immune environment, and host susceptibility to respiratory viruses in vitro. We will achieve this aim by assessing effect of nasal microbiome composition and nasal bacterial absolute abundance on: (i) innate immune defenses and (ii) influenza A, rhinovirus, and SARS-CoV-2 infectivity using an innovative in vitro nasal microbiome-respiratory epithelial co-culture model. The proposed research is innovative because it represents a departure from the status quo by assessing how the nasal microbiome, which is ubiquitous and diverse, could affect natural immune environment. significant because it is expected to identify nasal microbiome features and immune profiles that confer natural antiviral protection, which could inform novel strategies to reduce the risk for respiratory viral infections.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT Efficient wound healing requires complex cellular communication between tissue-resident non-immune cells and infiltrating immune cells. While much has been learned about how cytokines and growth factors contribute to acute wound healing, we know very little about how lipid signaling regulates acute inflammation and tissue repair, and identification of new mechanisms that govern acute inflammation and repair is needed. We previously demonstrated that inhibiting dermal adipocyte lipolysis led to reduced macrophage numbers during early inflammation and delayed repair, yet the mechanism(s) linking adipocyte lipolysis to efficient inflammation and repair have not been identified. Given the rising numbers of diabetic and aged patients, it is imperative to define molecular underpinnings that promote a healthy acute inflammatory response and to identify druggable mechanisms to treat inflammation and non- healing wounds. Inhibition of injury-induced dermal adipocyte lipolysis significantly reduces the abundance of medium-chain fatty acids (MCFAs). Recently, GPR84 was identified to be an MCFA receptor that is expressed by bone marrow-derived myeloid cells and during tissue inflammation. Activation of GPR84 in vitro increases macrophage migration and enhances pro-inflammatory gene expression; however, its role in skin and the in vivo mechanism of action is not well defined. We observe increased GPR84-expression during wound-induced inflammation and found that administration of a GPR84 agonist increases macrophage numbers. Additionally, systemic administration of a GPR84 antagonist decreases wound bed macrophages and delays tissue repair. Based on our preliminary data, we hypothesize that GPR84 signaling is required to support macrophage numbers and subsequent repair during injury-induced inflammation. We will combine our team’s tools and expertise in adipocyte, keratinocyte, and macrophage biology with single-cell data interrogation to validate this hypothesis with the following Specific Aims: (1) Use mouse models to determine how GPR84 signaling controls macrophage numbers and subsequent tissue repair after injury, and (2) define how MCFA/GPR84 signaling directly regulates myeloid cell function during skin wound healing and (3) define how epithelial GPR84 signaling contributes to keratinocyte function after injury. GPR84 signaling represents a new window to better understand mechanisms that regulate the injury response. Findings from this proposed work could lay a solid foundation for developing new tools that predict and enhance therapeutic treatment of wound healing.

NIH Research Projects · FY 2026 · 2023-02

Project Summary/Abstract Prostate organogenesis starts prior to birth, with extensive tissue growth occurring during puberty in response to androgens after the immune system is fully functional. Androgens are generally considered immune- suppressive, so this suggests contradictory roles for immune cells in the prostate, where an immune suppressive microenvironment is needed as new prostate-specific proteins are generated, yet immune cell functions are required to promote morphogenesis and tissue growth during puberty. Our recent single-cell RNA sequencing analyses of normal mouse and human prostates revealed previously unknown heterogeneity in prostate epithelial cell types. These studies form the foundation for a new direction in which I propose to shift my focus from prostate epithelial cells to investigate the function, developmental origins, and androgen-dependence of immune cells in the prostate. The long-term goal of this research is to investigate immune cell regulation of prostate organogenesis so that we can better understand the underpinnings of benign prostate disease. This application specifically seeks to elucidate the function of macrophages during prostate organogenesis and their potential contribution to BPH progression. Our central hypothesis is that heterogeneous populations of prostate macrophages with distinct cellular origins and tissue-specific properties regulate the organogenesis and function of the prostate, and their dysregulation contributes to BPH. To test our hypothesis, Specific Aim 1 will identify the function and heterogeneity of macrophages in the developing prostate. Specific Aim 2 will determine the cellular origins of macrophages in the developing prostate and in BPH. Specific Aim 3 will identify the androgen signaling requirements of macrophage populations in the developing prostate and in BPH. Under Aim 1, we will conduct immunofluorescence analysis of macrophages in prostate tissues, incorporating high-resolution 3D imaging, live imaging, and data from single-cell RNA sequencing. For Aim 2, we will conduct genetic-lineage tracing studies of macrophages of yolk sac, fetal liver, and bone marrow origin. For Aim 3, we will test the role of AR in prostate macrophages by deleting AR in macrophages during prostate organogenesis and in a mouse model of BPH. This proposal is innovative due to the novel investigation into how immune cells contribute to promoting prostate organogenesis both prior to and during puberty in this androgen-regulated organ; the novel investigation into how AR functions in prostate immune cells to regulate organ morphogenesis and immune suppression during puberty; and the use of innovative technology. The proposed research is significant because successful completion of this proposal will elucidate the in vivo function of macrophages during prostate organogenesis; demonstrate the cellular origin of prostate macrophages across the various stages of prostate organogenesis; and clarify the in vivo role of cell-autonomous AR in macrophages during prostate organogenesis. Importantly, this project will further investigate the role of macrophages in the development of BPH, which has the potential to inform the development of improved treatment strategies for BPH.

NIH Research Projects · FY 2026 · 2023-02

Low- and middle-income countries (LMICs) have high rates of tobacco use and secondhand smoke exposure (SHSe). Two countries particularly impacted by tobacco use and SHSe are Armenia (AM) and Georgia (GE), which represent the 11th and 6th highest smoking rates in men globally (51.5% and 55.5%). Notably, a primary source of SHSe among children and most nonsmoking adults in many LMICs, including AM and GE, is the home. Smoke-free homes (SFHs) can reduce SHSe, promote cessation, and possibly disrupt initiation; however, 61.4% of households in AM and GE allow smoking in the home. Thus, promoting SFHs may be an innovative and relatively untapped strategy for chronic disease prevention in these countries – and in other LMICs with high smoking rates. Research focused on implementing evidence-based interventions (EBIs) offers unique opportunities to address the pressing needs in LMICs and to examine key barriers in the adoption, scale-up, and sustainment of EBIs in low-resource settings. This proposal builds on our teams ongoing collaborations which have established: 1) a strong community-based infrastructure for implementing public health programs using local coalitions in 14 communities, in a prior Fogarty R01; and 2) a theory-based SFHs intervention, designed to be brief and adaptable and shown to be effective, generalizable, scalable, and cost-effective among low-income households in the US. This study will: (1) adapt our SFH intervention to be culturally appropriate for AM and GE, using a community-engaged approach and robust adaptation frameworks and methods, and develop in-country capacity for intervention dissemination (via local coalitions) and delivery (via national quitlines); (2) examine effectiveness of the adapted intervention (vs. control) on SFH adoption (primary outcome) among households in AM and GE, using a type 1 hybrid effectiveness-implementation RCT (n=550 participants), with follow-up assessments at 3 and 6 months; and (3) assess intervention reach, adoption, implementation, and maintenance potential, as well as related contextual influences, using a mixed-methods process evaluation. This work will provide a model for adapting and implementing this EBI for AM and GE, which could be used in other countries and/or for other behavioral targets and EBIs in AM, GE, and elsewhere.

NIH Research Projects · FY 2026 · 2023-01

Salt-sensitive (SS) individuals on high Na+ intake not only develop hypertension but also kidney injury/ chronic kidney disease (CKD) and cardiovascular disease (CVD). A reduction in Na+ intake may prevent and treat hypertension, CVD, and CKD. However, low Na+ intake may not always be beneficial in the treatment of hypertension or CVD. A low Na+ intake is associated with increased risk of hypertension (i.e., inverse salt sensitivity (ISS), CVD, and death. Hypertension and diabetes are the major causes of renal injury, accounting for up to 75% of end-stage renal disease. However, hypertension may cause CKD only in the genetically susceptible. In 13 of 16 studies in the GEO Dataset of CKD patients, dopamine type 2 receptor (D2R) gene (DRD2) expression is lower in those with CKD than those without CKD. A decrease in the expression or function of D2R, per se, or caused by DRD2 variants, increases renal inflammation, renal fibrosis, and ISS. The mechanisms/genetics of ISS are not well understood. Mice with global germline deletion of Drd2 (Drd2-/-) have SS hypertension and ISS. However, mice with renal proximal tubule (RPT)-specific conditional deletion of Drd2 (Drd2cPT) have increased blood pressure (BP) only when Na+ intake is decreased, a case of ISS. Sprague- Dawley rats have ISS, related to an increase in the activity of the angiotensin type 1 receptor (AT1R) and α1-adrenoceptors. About 15% of hypertensive subjects have ISS and some associated with DRD2 rs6276/rs6277. Renal-selective expression of DRD2 variant rs6277 in mice should increase BP and impair inhibition of renal Na+ transport and excretion. We will test the overall hypothesis that DRD2/Drd2 is important in preventing ISS by mitigating overly active renin-angiotensin and sympathetic nervous systems and the increase in RPT Na+ transport on a low Na+ diet. Specific Aim 1 will test the hypothesis that in Drd2-/- or Drd2cPT mice, BP increases when Na+ intake is “low”, a case of ISS. The increase in BP in Drd2-/- or Drd2cPT mice fed a low Na+ diet is caused by impaired D2R inhibition of RPT Na+ transport and an increase in RPT Na+ transport caused by activation of both the renin-angioten-sin and sympathetic nervous systems. In the long-term, renal function is decreased because of unmitigated renal fibrosis. Specific Aim 2 will test the hypothesis that DRD2 variants, rs6276/rs6277, decrease D2R expression that is dependent on the effects of the transcription factors NR4a2 and miR4301. These studies are significant and important because they may lead to the identification of the human population that would be adversely affected by the current recommendation to decrease the Na+ intake in everyone.

NIH Research Projects · FY 2026 · 2023-01

Project Summary Dynamic acetylation/deacetylation of histones and nonhistone proteins is a critical switch in gene regulation. Manipulation of the acetylation switch is emerging as a promising therapeutic strategy in the treatment of cancer. My laboratory has a long-term interest in clarifying the functions, mechanisms of action, and regulation of lysine acetyltransferases and deacetylases, exploring their roles in diseases, and using the resulting knowledge to develop new and better strategies for the treatment of diseases such as cancer. The key focus of this resubmission application is on the Males Absent on the First (MOF, also called KAT8 or MYST1) protein, a member of the MYST lysine acetyltransferase family. MOF regulates a variety of cellular processes including gene transcription, DNA damage responses, and embryonic development. The proposed project is significant because although increasing evidence suggests that MOF is also closely involved in cancer, the exact mechanism by which MOF impacts tumor development and progression is unclear. Our preliminary studies revealed an unexpected function of MOF in the transcriptional repression of epithelial to mesenchymal transition (EMT) and cytokine genes in lung cancer. Furthermore, MOF depletion significantly affects lung tumorigenesis in mouse models due to release of its transcriptional repression. Based on these exciting preliminary results, we hypothesize that MOF may be a potential target for treatments of lung cancer. The long-term objective is to explore how MOF regulates uncharacterized gene expression and signaling processes to impact lung cancer. The central hypothesis will be tested by pursuing three specific aims: 1) Dissect the novel and unexpected mechanisms by which MOF represses gene transcription; 2) Examine how MOF controls gene expression via the methyltransferase G9a and TGF-beta/SMAD; and 3) Explore the implications of SIRT1-mediated deacetylation of MOF in lung cancer development. The expected outcome of this work will help elucidate new functions of MOF and its mechanisms of action in lung tumorigenesis, and provide insights into future development of new therapeutic strategies for lung cancer.

NIH Research Projects · FY 2025 · 2022-09

Project summary/abstract The establishment of left-right (LR) asymmetry is a critical event required for the correct positioning of internal organs. Defects in human LR axis formation cause birth defects of the heart, vasculature, lungs, and intestinal tract. The gene network contributing to the generation of LR asymmetry is highly conserved across vertebrates. In the mouse, the initial asymmetric signals establishing LR axis are determined in the node by cilia-driven leftward fluid flow (nodal flow). These signals are then transferred to the left lateral plate mesoderm (LPM), which will undergo asymmetric organ morphogenesis. Nodal, a secretory protein that belongs to the transforming growth factor-b (TGF-b) superfamily, is expressed in the node and travels a long distance to the left LPM, where it initiates a transient auto-regulatory circuit (involving Nodal and Lefty) that propagates Nodal signaling and activates expression of the left-sided determinant Paired-Like Homeodomain transcription factor 2 (Pitx2). The Hedgehog (Hh) signaling pathway also plays a crucial role in LR patterning. In the mouse, Hh signaling is required to establish the midline that separates the left and right sides of the embryo, as well as to activate the Nodal-dependent auto-regulatory circuit in the LPM. However, the mechanism by which Hh signaling regulates the competence of the LPM for Nodal response is not well understood. Thus, identifying the specific convergence point of the Hh and TGF-b pathways is critical for understanding the in-depth mechanism underlying LR asymmetry determination, and therefore to provide better diagnosis, preventive and therapeutic intervention against LR asymmetry-related birth defects. We recently found that a novel target of the Hh pathway, Neuron-Derived Neurotrophic Factor (Ndnf), regulates axial rotation and intestinal looping in the mouse. In the proposed work, we will incorporate biochemical, molecular, and genetic approaches to (Aim 1) assess the source, (Aim 2) determine the transcriptional regulation, and (Aim 3) investigate the underlying molecular mechanisms of Ndnf in LR determination.