Ohio State University

NIH Research Projects · FY 2025 · 2023-07

Severe bacterial infections are a major cause of global mortality and morbidity. An aberrant host response to infection leads to destructive inflammation and extensive tissue damage, resulting in organ dysfunction and multi- organ failure. An increasing incidence of gram-negative bacteria (GNB) resistance to antibiotics has been associated with increased mortality and significant public health problems in the world over last two decades. Uncontrollable inflammation is a critical feature of GNB pneumonia-induced acute respiratory distress syndrome (ARDS), a devastating complication of severe sepsis. Since there are no specific treatment available, current research focuses on identifying new drug targets to diminish pro-inflammatory responses. Dysfunction of protein homeostasis in immune system has been known to contribute to the pathogenesis of systemic inflammation. Protein degradation is mainly controlled by the proteasome. Recently, a specific subgroup of the proteasome called the immunoproteasome has been identified to play a critical role in inflammatory responses including antigen presentation. In the preliminary study, we found that endotoxin increases immunoproteasome structural assembly; however, the molecular regulation of immunoproteasome structural assembly and its role in the pathogenesis of ARDS have not been revealed. We hypothesize that deubiquitinase USP14 determines immunoproteasome structural assembly, and that inhibition of the immunoproteasome diminishes NLPR3 inflammasome activation and GNB-induced inflammation. We will determine molecular mechanisms by which USP14 activation regulates LPS-induced immunoproteasome structural assembly. Next, we will determine the mechanisms underlying how the phosphorylation of USP14 by PKCδ promotes immunoproteasome structural assembly and severity of GNB-induced lung injury and sepsis. We will use state-of the art molecular approaches, human samples, and preclinical animal models. The data will lay the foundation for a significant mechanistic advancement regarding the molecular regulation of the pro-inflammatory responses through the modulation of the inducible immunoproteasome structural assembly, which are implicated in the pathogenesis of acute bacterial infection.

NIH Research Projects · FY 2026 · 2023-07

ABSTRACT Despite the fact that multiple different treatments for depression have been available for decades, the global burden of the illness has grown steadily. Depression is now one of the leading causes of disability worldwide. Current treatment strategies for depression remain largely trial-and-error, and fewer than 40% of patients respond to a given treatment and sustain that response for a year, even when treatment is continued. A central barrier to improving these outcomes is the need to characterize better phenotypes of depressive illness that are more closely aligned to modifiable neurobiological targets than are current symptom constellations and diagnostic codes. Findings from our group, and from others, suggest that one such phenotype involves the propensity to experience anger, hostility, and irritability following negative experiences and to respond in an aggressive, overly hostile manner (hereafter denoted Angry Hostility). Our preliminary data suggest that the Angry Hostility phenotype is associated with a particular pattern of altered functioning in neural regions that support emotion processing and emotion regulation. Furthermore, Angry Hostility appears to be strongly associated with hostile, aggressive behaviors following provocation and with real-world interpersonal and work- functioning impairments that can exacerbate depressive symptoms. The primary goal of this project is to test a novel model through which higher levels of Angry Hostility among adults with depression are associated with specific patterns of abnormal neural function and behavior, leading to poor functional outcomes and future symptoms. To achieve these goals, 150 adults (18-45 years old) with at least mild symptoms of depression will be recruited, as will 100 demographically matched, psychiatrically healthy individuals. Participants will complete clinical, neuroimaging, and laboratory behavioral assessments, as well as 4-, 8-, and 12-month follow-up assessments and four 10-day ecological momentary assessment protocols. The project will examine 1) whether Angry Hostility is associated with abnormal neural function in emotion processing and emotion regulation regions; 2) whether Angry Hostility is associated with aggressive behaviors in the laboratory and in real-world settings; and 3) whether abnormalities in a-priori neural systems and behaviors prospectively predict poorer real- world functioning and psychiatric symptoms over the 12-month follow-up. The aims of the project match well with the strategic goals of the National Institute of Mental Health. Moreover, the results of this study have the potential to describe the neurobiological bases, behavioral mechanisms, and real-world consequences of elevated Angry Hostility among adults with depression. Future work will aim to develop personalized treatments to target the neural mechanisms identified in this study in order to reduce symptoms and improve functional outcomes for adults with depression who have higher levels of Angry Hostility.

NIH Research Projects · FY 2025 · 2023-06

Lung cancer is highly lethal and accounts for nearly as many deaths as breast, cervical, colorectal, and prostate cancers combined, but most lung cancer cases are potentially preventable. About 80-90% of cases are smoking-related, and screening followed by high-quality treatment has been shown to reduce the risk of death in people at high-risk. Therefore, smoking cessation interventions plus screening are two complementary pillars that are recommended together by the US Preventive Services Task Force and other guidelines to reduce the lung cancer death rate. Unfortunately, uptake of lung cancer screening (LCS) remains low and although differences in lung cancer death rates by socioeconomic factors, including rurality, are widely acknowledged, they remain understudied. Evidence shows that rural areas have higher lung cancer death rates than urban populations, likely related to differences in risk factors and healthcare access. However, efforts to understand and address rural-urban outcome differences are hampered by low representation of rural populations in public data systems. The goal of this proposal is to elucidate factors that contribute to rural-urban differences in lung cancer deaths and provide contextual information for future interventions and policies. Our specific aims are to: (1) Characterize the delivery of evidence-based interventions (EBIs) for lung cancer prevention and early detection, comparing rural to urban areas, by assessing differences in use of smoking cessation interventions and LCS at multiple levels of influences; (2) Identify potentially modifiable care gaps across the LCS continuum, including risk assessment and timely treatment, by examining patients who died of lung cancer relative to patients who are alive by rural-urban status; and (3) Evaluate similarities and differences in the barriers and promoters to delivery of EBIs across the LCS continuum (smoking cessation, shared decision-making, screening, and treatment) in the rural and urban contexts. We will use a multidimensional health outcome heterogeneity framework to apply a convergent, mixed methods approach for our studies. We will leverage the Rochester Epidemiology Project, a unique population-based data resource for a 27-county contiguous area in the Midwestern US, along with the Southern Community Cohort Study across 12 states in the Southeastern US, among a population of 50-80-year-old people. We will use Rural-Urban Commuting Area codes to define rurality and will assess both self-reported and area-level socioeconomic information. We will conduct semi-structured interviews with patients and clinical staff to gain perspectives on LCS barriers and promoters, including the potential role of smoking-related stigma. The proposed research has the potential for high impact by elucidating gaps on lung cancer prevention and early detection that will translate directly into strategies to address intransigent heterogeneity in outcomes across our study populations, and beyond. We will thus address priorities of the NCI and the President’s Cancer Panel to advance improvements in the delivery of lung cancer prevention and early detection. Our transdisciplinary team has a track record of high-impact research and has the expertise needed to successfully complete this research.

NIH Research Projects · FY 2025 · 2023-06

Project Summary Breast cancer has been the most prevalent cancer and the second leading cause of cancer-related death in American women for many years. Immune checkpoint inhibitors (ICIs) targeting checkpoint proteins such as programmed cell death protein 1 (PD1) resulted in durable clinical remissions in a subset of cancer patients, including breast cancer. However, most patients didn’t show a response to ICI treatment, urging the need for novel biomarkers that can predict patient response and therapeutic targets that can improve the efficacy and durability of ICIs. The goal of this proposal is to investigate how to overcome the ICI resistance. My preliminary data showed that interferon (IFN) -alpha and -gamma signaling are enriched in the tumor and blood neutrophils of nonresponders to ICIs. The central hypothesis of this proposal is that tumor-educated neutrophils with increased IFN signaling mediate breast cancer resistance to ICIs, and can be used as predictive biomarkers. During the K99 phase, I will explore the impact of neutrophil-restricted IFN signaling on tumor response to ICIs and characterize the neutrophil-specific interferon-stimulated gene (ISG) signature (Aim 1). Since we found that ISGs in peripheral blood neutrophils can predict breast cancer response to anti-PD1 therapy, I will determine if blood neutrophil ISGs signature can serve as a biomarker in other cancer types and human patient samples from the clinic and clinical trials (Aim 2, K99 and R00). Finally, I will study the neutrophil IFN signaling in regulating immune memory and durable response to ICIs (Aim 3, R00). Upon successful completion of the Specific Aims, this translational study will extend our knowledge of neutrophil IFN signaling and provide novel biomarkers for the ICI therapy and therapeutic targets to overcome the ICI resistance. My overall career goal is to establish an independent translational cancer research group that will improve understanding of cancer development, identify novel effective therapies, and train the next generation of cancer researchers. This proposed research in the K99 phase will take place in the Lester and Sue Smith Breast Center at Baylor College of Medicine, a highly collaborative and multidisciplinary environment with strong integration of basic, translational, and clinical research. The institution is dedicated to the career development of postdoctoral trainees, and provides a variety of training venues including bioinformatics and immunology, weekly seminars, R&D workshops, journal clubs, and the annual retreat. BCM is part of the Texas Medical Center, the largest medical city in the world consisting of over 60 medical institutions and hospitals, which offers me enormous opportunities for training and collaboration. Finally, I will meet with Drs. Rosen and Zhang weekly to discuss my projects besides our weekly lab meetings and have a formal committee meeting every three months to discuss my progress and receive feedback. I am also supported by a patient advocate and other collaborators. Through the training and research plan proposed within my K99/R00 application, I will acquire knowledge and skills which will greatly improve my ability to launch my scientific career as an independent investigator.

NIH Research Projects · FY 2026 · 2023-06

Retroviruses are obligate intracellular parasites that must integrate a copy of their viral genome (cDNA) into a host chromosome. Integration is accomplished by the retrovirus-encoded integrase (IN) that forms a catalytic complex with two viral cDNA long terminal repeat (LTR) ends, termed an intasome. Retroviral intasomes maintain a conserved intasome core that may be expanded into higher order IN multimer architectures. For example, the prototype foamy virus (PFV) intasome is a simple IN tetramer, while the mouse mammary tumor virus (MMTV) and Rous sarcoma virus (RSV) intasomes are IN octamers. Even higher IN multimers have been reported for the lentiviruses that include HIV-1 and Maedi-Visna virus (MVV). While numerous biochemical and cellular studies have detailed retroviral integration, the assembly mechanics and cost-benefit of different multimeric IN architecture on intasome biophysical properties is a substantial knowledge gap in retrovirology. Our previous work detailed the dynamic target search, integration kinetics, DNA lesion interactions, IN domain requirements and nucleosome targeting by PFV intasomes. Real-time single molecule studies were also performed with MMTV intasomes. Several important differences were identified between the PFV tetramer and MMTV octamer intasomes including distinct target search and strand transfer kinetics as well as the ability of MMTV to form multivalent complexes on a target DNA. These observations have prompted several key questions: What are the contributors that determine IN multimeric architecture? What are the factors of IN multimeric architecture that influence target search and strand transfer? How does intasome architecture influence chromatin DNA binding and target site selection? The PFV, MMTV, RSV and MVV intasomes are convenient biophysical models for probing intasome architecture since they naturally exist as an IN tetramer, octamer or 16-mer with published structures and assembly protocols. We have found that swapping the non-conserved peptides that link the signature conserved retroviral IN protein N-terminal domain (NTD), catalytic core domain (CCD) and C-terminal domain (CTD), converts them into active intasomes with a multimeric architecture of that often mimics the donor intasome. How and why these non- conserved linker peptides influence intasome architecture is unknown. We propose to utilize multiple highly quantitative single molecule imaging tools to understand the contributions of IN multimeric architecture on retroviral mechanics with the following Specific Aims: 1.) examine IN-multimer assembly and integrase activities that distinguish intasome architectures, 2.) determine the role of intasome architecture on the dynamic interactions with defined duplex and chromatin target DNA, and 3.) determine the role of intasome architecture on targeting host chromatin features in vivo. These studies will interrogate the contributors to IN multimer architecture and intasome dynamics with the goal of identifying new retroviral mechanics and therapeutic targets.

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY Cancer and chronic virus infections are significant causes of morbidity and mortality. While cytotoxic CD8 T cells are the main killers of tumors or virus-infected cells, persistent stimulation of CD8 T cells during chronic infections or cancer results in a gradual loss of their cytotoxic function as T cells progress towards a fully- exhausted state. While immune checkpoint blockade (ICB) therapy allows partially-exhausted CD8 T cells to functionally recover by blocking inhibitory signals, terminally-exhausted T cells remain nonresponsive to this therapy. The inability of terminally-exhausted T cells to recover after ICB may explain why many cancer patients fail to mount durable responses to ICB. We recently showed that de novo DNA methylation programming is causally linked to the progression of T cells toward terminal exhaustion and poor response to ICB. Importantly, we discovered that targeting T cell-intrinsic epigenetic programs synergized the efficacy of ICB during chronic infection or cancer. Yet, the following questions represent major gaps in our current understanding of T cell exhaustion: (1) How are these epigenetic changes acquired in exhausted T cells? (2) What are the upstream signals that regulate the specificity of de novo DNA methylation programs in exhausted versus functional T cells? (3) Can we reverse the epigenetic programming in exhausted T cells to the functional state while avoiding transformation? Bridging these gaps will allow us to identify and target factors that apply “epigenetic brakes” to CD8 T cell function. To address these questions, we have developed a novel in-vitro model of stable human T cell dysfunction as a tractable tool that can guide our in-vivo experiments by providing first-line mechanistic studies. First, we will employ cutting-edge approaches, such as CRISPR-Cas9 gene editing and retroviral transduction, to test the hypothesis that specific tumor microenvironmental signals regulate epigenetic programming in persistently stimulated CD8 T cells, which promotes their resistance to ICB therapy. We aim to block and/or revert the progression toward the terminally-exhausted state by targeting components of this signaling pathway while promoting counteracting pathways. Second, we aim to rebalance specific microenvironmental signals to restore functionality and response in terminally-exhausted T cells. Using novel in- vitro model systems of T cell dysfunction and complementary in-vivo models of chronic viral infection and cancer, as well as cutting-edge technologies to profile DNA methylation, open chromatin landscape, and transcriptome in CD8 T cells, our proposed studies can determine if targeting specific factors can remodel chromatin back into an accessible state at effector and/or stemness-associated genes, leading to functionally-rejuvenated T cells. These proposed studies will provide insights into how epigenetic programming can be reversed during progression to T cell exhaustion that can be translated to reprogram terminally-exhausted T cells in clinical settings, ultimately enhancing the efficacy of T cell immunotherapies.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY Atherosclerotic cardiovascular disease (ASCVD) is an inflammatory disease of unclear pathogenesis that remains the leading cause of morbidity and mortality throughout the world. Immune responses play a central role in the evolution of this chronic disease, and CD4+CD25+Foxp3+ regulatory T cells (Tregs) exhibit critical functions to regulate inflammation. We have shown the level of expression of CD39, an immunosuppressive ectonucleotidase, on Tregs to be genetically associated in humans with specific single nucleotide polymorphisms (SNPs). Although select SNPs for CD39 have been linked to Crohn disease, no studies have examined whether alterations in Treg CD39 catalytic activity, and changes in extracellular ATP scavenging with adenosine generation, impact manifestations of ASCVD. Similarly, a naturally occurring antisense ENTPD1-AS1 has been shown to decrease CD39 expression in Crohn’s patient Tregs. Inhibition of ENTPD1-AS1 with a specific self- delivering FANA CD39 antisense (FANA-CD39-AS) oligonucleotide were shown to increase CD39 expression, providing a therapeutic option to modulate Treg phenotype. Our overall experimental goal is to elucidate the impact of genetic regulation of Treg CD39 expression in ASCVD. The central hypothesis is that genetic mutations resulting in decreased Treg CD39 activity and altered purinergic responses drive ASCVD due to the inability to adequately resolve inflammation. We will test this hypothesis by conducting functional genomic experiments, in addition to using novel murine experimental model systems, and develop clinical studies, examining isolated cells and patient biospecimens. The proposal consists of two Aims. In SA1, our investigative team will determine how Treg expression of CD39 activity impacts atherosclerosis in a validated experimental system. This will be done using CD39 Treg-specific conditional knock-down and knockout murine models with multiple readouts of T cell and myeloid activation responses, and adoptive transfer studies, In SA2, we will examine the regulation of CD39 expression on human Tregs and examine the impact of CD39 modulation on immune function and inflammation in clinical studies of ASCVD. We have assembled a collaborative team with clinical and experimental expertise in ASCVD, immunology, vascular biology, genetics, and biostatistics. Completion of the proposed aims will develop understanding of the role of Tregs, and specifically expression of CD39 and altered purinergic responses, in ASCVD, this most important and significant disease. Translation to clinical practice will be facilitated by identification of important biomarkers and novel targets, inclusive of CD39 and related pathways of adenosinergic signaling, for therapeutic intervention in ASCVD.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY: Cardiac fibrosis is regulated by the activation and phenotypic switching of quiescent cardiac fibroblasts (CFs) to active myofibroblasts (MFs) which have extracellular matrix (ECM) remodeling and contractile functions which play a central role in cardiac remodeling in response to injury. As such, there is consensus effort in the field to manipulate fibroblast activity for therapeutic gain. However, a more complete understanding of the signaling pathways and mechanisms that regulate MF activity in cardiac remodeling remains an unmet need. We previously demonstrated that the RNA binding protein Human antigen R (HuR) directly mediates hypertrophic signaling in cardiac myocytes (CMs), and that CM-specific genetic deletion or pharmacological inhibition of HuR reduces pathological remodeling and preserves cardiac function following transverse aortic constriction (TAC)-induced pressure overload in part through a reduction in pro-fibrotic gene expression. New preliminary data suggests that HuR activity in cardiac fibroblasts may play an equally important role in cardiac remodeling. Our new data demonstrates a necessary role for HuR in MF activation and the ECM-remodeling capacity of cardiac fibroblasts. Furthermore, we have identified Wisp1 (Ccn4) as a downstream HuR- dependent mediator of MF activation, and show that exogenous addition of recombinant Wisp1 partially restores MF activity upon HuR inhibition. The primary goal of this proposal is to determine the functional role that HuR-Wisp1 signaling plays on MF activity and whether functional inhibition of these pathways in fibroblasts will provide therapeutic benefit during pathological cardiac remodeling. Our central hypothesis is that HuR-Wisp1 signaling in cardiac fibroblasts is necessary for myofibroblast activity and promotes pathological cardiac remodeling. The specific Aims of this proposal are to: (1) Determine the functional role of HuR in CFs in vivo and define its pleiotropic role across cell types during pathological cardiac remodeling. (2) Identify the functional and mechanistic role of HuR-dependent control of Wisp1 expression on MF activity and pathological cardiac remodeling. The expected results of this proposal will provide a deeper understanding of the functional impact of the HuR-Wisp1 signaling axis across cardiac cell types during pathological cardiac remodeling that is necessary for potential therapeutic manipulation of HuR or HuR-dependent gene expression in cardiac remodeling as suggested by our previous work.

NIH Research Projects · FY 2025 · 2023-05

PROJECT SUMMARY The gut microbiota has been linked to many aspects of human health and disease. These finding have ignited efforts to precisely modulate gut microbiota composition and function to promote health-associated features. The nutrient landscape within the gut shapes, and is influenced by, the gut microbiota. Bacteria respond to available nutrients and utilize them to support their own metabolism, sharing the metabolic by-products with other bacteria and the host. Carbohydrates within the gut, both consumed in the diet and produced by the host, impact gut bacteria composition and function via their utilization as a carbon source. The biological function of the polysaccharides that cover gut bacteria however, remains unclear. Bacterial cell surface polysaccharides act as a barrier between the microbe and its environment, enhancing bacterial growth and survival through mechanisms that include resistance to toxic small molecules, nutrient adaptation, and immune evasion. The central hypothesis I will test in this proposal is that microbiota bacterial polysaccharides modulate gut community structure and function via utilization as a nutrient by other community members, and through interaction with soluble immunoregulatory proteins. AIM 1 will employ isolated bacterial polysaccharides and in vitro growth assays to identify genetic features that enable utilization of bacterial polysaccharides. AIM 2 will define whether bacterial polysaccharides are consumed in vivo by cultured, genome sequenced microbial communities installed in gnotobiotic mice using microscopic recoverable paramagnetic beads coated in polysaccharides. AIM3 will test whether cell surface polysaccharides from probiotic dietary supplements alter gut microbiota polysaccharide utilization and recognition of community members by immunoregulatory proteins in the gut lumen of gnotobiotic mice. This series of experiments that blends chemistry, glycobiology, genomics, and gnotobiotic mouse models will define mechanisms of bacterial polysaccharide utilization, increase understanding of how nutrients in the gut shape the microbiota, and suggest a bioactive component of bacterial dietary supplements. These combined finding should improve development of microbiota-derived and -directed therapeutics for targeted microbiota manipulation. This award will also support by career development. During completion of the supervised portion of this grant I will gain critical computational research skills that includes bacterial genome sequencing and annotation, bacterial RNA-sequencing to characterize function, and metagenomic analysis. Ultimately, this award will facilitate my successful transition into an independent academic position at a research-intensive university where I will lead, teach, and mentor an interdisciplinary group of students, postdocs, and clinicians defining mechanisms of microbiota assembly, function, and regulation with a goal to translate my findings into methods for targeted microbiota manipulation to improve human health.

NIH Research Projects · FY 2025 · 2023-05

PROJECT SUMMARY/ABSTRACT Obesity and type 2 diabetes are increasing dramatically worldwide. Recent data has shown that obesity rates have more than doubled since 1980 and if these trends continue unabated, by 2030 estimates predict that approximately half the U.S. population will be obese, with 25% developing type 2 diabetes. Adipose tissue is an important site for initiation and aggravation of obesity and type 2 diabetes because it is a key endocrine organ that functions to maintain energy homeostasis, and recent studies have identified lipokines, or signaling lipids, released from adipose tissue as molecules that can mediate metabolic effects. One class of these lipokines are oxylipins, oxidized lipid metabolites that exert metabolic effects. An important oxylipin for metabolic control is the linoleic acid metabolite 12,13-diHOME, which is released from brown adipose tissue (BAT) in response to cold and exercise in rodents and humans and beneficially impacts glucose and fatty acid metabolism. 12,13-diHOME increases fatty acid uptake into brown adipose tissue, skeletal muscle, and cardiomyocytes, and is negatively correlated with circulating triglycerides and BMI in humans. However, due to its very short half-life, systemic regulation is difficult to maintain and thus its therapeutic potential has not been fully realized. To address this essential issue, we developed a paradigm-shifting approach to increase 12,13- diHOME via tissue nanotransfection (TNT), a non-viral gene delivery technology with high translational potential. TNT delivery of the genes Ephx1/2, coding for the enzymes that make bioactive 12,13-diHOME results in a sustained systemic increase of 12,13-diHOME in circulation and corresponds to reduced adiposity and improved metabolic health. In these proposed studies we will optimize a therapeutic upregulation of Ephx1/2 via TNT in pre-clinical models and comprehensively establish the physiological ramifications of a sustained systemic increase in 12,13-diHOME and provide new therapeutic approaches to combat obesity and type 2 diabetes. We will do this using the following two specific aims: 1) Determine the effectiveness, efficiency, and mechanisms through which TNT-based delivery of Ephx1/2 into the skin increases 12,13- diHOME in circulation and; 2) Determine the physiological ramifications of a sustained increase in the oxylipin 12,13-diHOME by TNT. The proposed studies have the potential to provide paradigm-shifting results and elucidate novel mechanisms to sustain oxylipin up-regulation and providing new therapeutic approaches to combat obesity and type 2 diabetes.

NIH Research Projects · FY 2025 · 2023-05

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths among both men and women in the US, with screening recommended by the US Preventive Services Task Force (USPSTF) to reduce incidence and mortality. Despite national targets, CRC screening rates, especially among patients receiving services in low-resourced settings, remain below these goals. Federally Qualified Health Centers (FQHCs) primarily serve people from low socioeconomic status (SES) groups and have some of the lowest screening rates. Health-related social needs (HRSNs), such as housing instability and transportation issues, play a significant role in CRC screening completion disparities. However, their impact on patients' CRC screening completion remains underexplored. This administrative supplement proposal will investigate the associations between HRSNs and follow-up colonoscopy receipt among FQHC patients with abnormal stool-based test results. This population will be reached by leveraging outreach to safety-net settings such as The Ohio State University Wexner Medical Center, KPNC/KPSC and the American Academy of Family Physicians National Research Network partnership—all strategies for aim 3 of the parent grant. This supplement complements the proposed work in the parent grant, primarily through its support of its first specific aim. For parent grant Aim 1, the supplement will contribute a qualitative approach to understanding the CRC screening continuum through the lens of a particularly low-SES population (FQHC patients), paying special attention to patients' HRSNs. The supplement will further contribute to the understanding of recommendation-concordant CRC treatment timeline adherence for members of this population who receive abnormal stool test results. The results of the supplement will provide covariates of interest (e.g., specific HRSNs or social determinants of health) for the statistical analyses proposed in Aims 2 and 4 of the parent grant. Data will be collected that may ultimately influence the development of interventions targeting HRSNs to enhance CRC screening completion rates in populations in low SES. Drawing from existing literature on other cancer types, we hypothesize that patient-level HRSNs strongly influence screening completion and subsequent outcomes. Dr. Aldenise Ewing, the DS candidate, aspires to become an independent investigator focusing on addressing cancer-related health disparities. This supplemental application will provide a mentored experience in cancer health disparities and cancer control for Dr. Aldenise Ewing. The study findings will provide preliminary data on social determinants of CRC-related outcomes and strengthen her K01 submission within 2 years of funding and R01 or equivalent grant submission within 5 years of funding. Moreover, it will contribute crucial insights into addressing social determinants of health in low-resource settings, ultimately improving health outcomes for all populations.

NIH Research Projects · FY 2026 · 2023-05

Summary Drug-drug interactions (DDIs) and pharmacogenetics (PG) are leading causes of adverse drug events (ADEs), with one in four patients experiencing ADEs attributable to DDIs or PG. However, despite the intrinsic connection of their pharmacological mechanisms, DDI and PG are often studied separately. There is a significant need for more efficient and effective translational from DDI to PG research, and newly developed machine-learning (ML) and artificial-intelligence (AI) methods have made such research feasible. In our recent DDI knowledge-discovery study of 25 million PubMed abstracts, we used ML and natural-language-processing analyses for the first time to identify 986 DDI pairs with overlapping pharmacokinetic mechanisms and clinical evidence, from which we generated 137 new PG hypotheses regarding CYP2D6 and CYP3A. In this grant proposal, we will develop novel ML methods, including active learning that will allow human annotator involvement and knowledge base reasoning that relies on logical rules to represent pharmacological mechanisms. This proposal has three aims: (1) to develop an active-learning approach to perform DDI and PG information retrieval analysis from the literature; (2) to develop a joint information-extraction and knowledge- base-reasoning approach to perform DDI and PG information extraction analysis from the literature; and (3) (a) to examine whether CYP3A/CYP2C19 genetic polymorphisms are associated with omeprazole-induced myopathy, and (b) to develop a prioritization scheme to examine new PG hypotheses generated from the literature-based discovery analyses from Aims 1 and 2 using Vanderbilt University’s BioVU biobank. These PG findings will provide a valuable resource for the wider scientific community for potential prospective studies and contribute significantly to the improvement of precision medicine and clinical care.

NIH Research Projects · FY 2026 · 2023-05

While alcohol remains the most widely endorsed substance during adolescence, and various psychosocial risk factors have been identified, unknown are the potential origins for how differences in alcohol related problems emerge among young adults. Applying a bio-psycho-social model to youth alcohol use (AU) prevention research is necessary to identify high impact points of intervention during adolescence before AU related problems emerge in adulthood. Given that both alcohol use and mental health distress emerge in adolescence, investigating relationships between bio-psycho-social factors and AU and related problems in adolescence is critical. We will consider both the risk and protective role of multilevel bio-psycho-social risk and protective factors for AU and mental health during adolescence using the ecodevelopmental framework, including Hispanic youth. Further, understanding bio-psycho-social risk and protective factors in association with AU and related problems will guide interventions that can target multiple bio-psycho-social factors for youth. This proposal will leverage the Adolescent Brain Cognitive Development® (ABCD) Study, a large comprehensive dataset, to advance critical areas of research in AU prevention among youth. A quantitative analysis will test (1) bio-psycho-social factors of AU related problems and mental health, and (2) AU influences on mental (anxiety/depression) in association with cognitive and school performance in youth. This proposal will use ABCD study longitudinal data to identify promotive factors that may buffer risk for AU and related problems influenced by both risk from individual and broader contextual factors. Potential co-emerging differences in mental health among youth will be investigated, and whether mental health (i.e., anxiety/depression) outcomes moderate (i.e., exacerbates) the influence of AU on adolescent cognitive functioning. This proposal will then extend the application of the bio-psycho-social model with community-engaged research approach to conduct a qualitative study with focus groups with Hispanic youth and parents to obtain a response to the ABCD study quantitative findings. Findings will inform hypotheses on future AU prevention research using community-engaged approaches with youth in a future R01. In summary, the quantitative and qualitative studies proposed will investigate the bio-psycho-social factors of risk and protective factors for alcohol use and mental health and subsequent impact on cognitive development. Findings will inform future interventions on AU prevention on timing and the interplay of multilevel bio-psycho-social factors.

NIH Research Projects · FY 2026 · 2023-05

Targeting microglia to alleviate Alzheimer’s Disease pathobiology Summary Alzheimer’s disease (AD) is the most common cause of age-related dementia leading to irreversible neurodegeneration and cognitive decline with no cure or effective preventive measures. Autophagy is a conserved, cell response found in all eukaryotic cells. Several studies in AD showed that autophagy activity is compromised in neuronal cells and suggested that this leads to reduced clearance of amyloid-β (Aβ) and neurotoxicity. Few reports examined autophagy activity in the AD brain and suggested that autophagy is dysfunctional in neurons, however, the contribution of autophagy in microglia, the immune cell of the brain, is unclear. Our newly generated data that was recently published show that autophagy is impaired in adult microglia from AD mice. More importantly, we found that a specific microRNA (miRNA), that targets several autophagy molecules, is upregulated in the brain of AD patients when compared to non-AD individuals, as well as in the brain and microglia-derived from an AD mouse model. Increased expression of this specific miRNA in the AD brain leads to down-regulation of autophagy effectors, which is then responsible for reduced clearance of Aβ by microglia. In Aim 1, we will determine the mechanism underlying elevated expression of the miR. In Aim 2, we will investigate the functional consequences of reducing this miR in the brain of AD mouse and the mechanism by which the microRNA is upregulated in AD brain. We will determine the behavior of microglia isolated from treated mice. These experiments will be performed using the AD mouse model 5XFAD and human samples from AD and non-AD patients. Our proposal will characterize a novel drug target and mode of delivery for AD.

NIH Research Projects · FY 2026 · 2023-05

Project Summary Prostate cancer (PCa) is the most frequently diagnosed male cancer and the second leading cause of cancer deaths in men in the United States. This growing public health challenge is aggravated by disparities in the incidence and mortality of PCa between African-American (AA) and European-American (EA) men. For example, the incidence of PCa is almost 60% higher in AA men and the mortality rate 2-3 times greater. While access to medical care may contribute to these differences, other studies suggest that cell-based differences may play a critical role. Unfortunately, no primary human prostate cell cultures are available for interrogating potential cellular alterations during early carcinogenesis. Organoid culture models work well for growing normal prostate cells and advanced PCa, but fail to succeed with primary PCa. CR (Conditional Reprogramming) culture, which was developed by Dr. Liu (PI) and his colleagues, is changing the landscape for generating in vitro human cancer models. CR technology allows to establish cell cultures from normal prostate, primary PCa and advanced PCa. CR cells from normal epithelium can fully differentiate when placed in conditions that mimic their natural environment, while CR cells from a primary prostate tumor exhibit an abnormal karyotype and form tumors in SCID mice. In the current application, in an effort to define the biological basis for their clinical disparities, we first propose to probe primary AA and EA normal prostate cells for differences in their susceptibility to immortalization and transformation. Then, we will determine response of biobanked normal and tumor CR cells to testosterone from AA and EA patients in presence or absence of their corresponding fibroblasts. Then, we will compare the genetic and biological properties of tumor CR cultures from AA and EA patients, including migration/invasion, anchorage-independent growth, tumor formation in presence or absence of their corresponding fibroblasts. Finally, we will compare genetically, epigenetically and phenotypically CR cells from AA and EA patients with metastatic and castration resistant PCa. Upon completion of this application, we will have established a living biobank with novel functional cell models, including matched normal and tumor prostate CR cells and their corresponding fibroblasts from AA and EA patients, immortalized AA and EA prostate cell lines and transformed AA and EA cell lines with annotated genomic and patient’s clinical information. These novel models include prostate cells at normal, primary PCa and advanced PCa and will provide an invaluable and novel resource for studies of initiation and progression and health disparity studies of PCa.

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY: Studies critically examining the cost-benefit of electronic cigarette (EC) flavors on smoking are urgently needed as FDA is making sweeping and impactful regulatory decisions without this important information. Surprisingly, no independent, long-term, randomized clinical trial has experimentally examined the impact of non-tobacco flavored vs. tobacco flavored ECs on product uptake, sustained use, and complete switching. While the National Academies of Science, Engineering, and Medicine concluded that ECs are likely to be far less harmful than combustible cigarettes, more than 1 million EC products have been banned since 2020, significantly narrowing the marketplace. According to the US FDA, a key reason for the removal of these products was the significant rise in youth vaping, often with flavored ECs. Indeed, the FDA decisions cited a failure of manufacturers to provide sufficient evidence demonstrating that the benefits to adult smokers outweighed the “documented risks to youth,” suggesting that the significant rise in youth vaping significantly tipped the scales a priori in the direction of restricting EC flavors. The current restricted legal EC marketplace may reduce youth vaping but may inadvertently reduce the ability of ECs to compete successfully with conventional cigarettes, reducing adult smokers’ trial and use of e-cigarettes for switching. FDA requires more robust and definitive studies evaluating the benefit of EC flavors, if any, to adult smokers. To provide the needed scientific evidence, we propose a nationwide randomized clinical trial to determine the impact of EC flavors on 1) product uptake and appeal, 2) cigarette craving, symptoms, and dependence, and 3) smoking behavior, including sustained and complete switching from cigarettes to ECs. We will also utilize combination nicotine replacement therapy (NRT, patch and lozenge) as an FDA-approved comparator to determine the potential increased benefit (or not) of EC vs NRT on tobacco use behavior. Smokers (N=1,500) will be randomized to a) preferred flavor EC (PEC); b) tobacco flavor EC (TEC); or c) combination NRT. Products will be provided at no cost for 14 weeks (2-week trial before switch date, 12 weeks of use following switch date). PEC participants will be provided their preferred flavor(s) and able to change flavors throughout the 14 weeks. Changes in smoking will be biochemically confirmed via remote exhaled carbon monoxide reading at 12 weeks (end of product provision) and 26 weeks after the switch date. This significant and innovative study will be the first to provide the FDA with critical and definitive information as to the impact of EC flavors on tobacco use among adult smokers. With expertise conducting nationwide, large- scale EC and NRT trial research, our team is uniquely suited to conduct this investigation. If our study demonstrates no significant improvements in switching with flavored EC use, then the continued sale of these products is likely indefensible; however, if improvements are significant, these findings will provide a critical counterweight to the current FDA regulations and aid future decision making.

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY This proposal focuses on elucidation of the processes regulating human natural killer (NK) cell development in the healthy state and in the setting of uterine endometrial carcinoma (EC). NK cells are cytotoxic “group 1” innate lymphoid cells (ILCs) that play myriad roles in immunity and are vital to controlling malignant transformation. NK cells undergo terminal differentiation and maturation in various tissues throughout the body, leading to a broad spectrum of NK cell phenotypes and functions. So-called conventional NK cells (cNK) that arise in secondary lymphoid tissues and predominate in the blood have established roles in complementing T cell-mediated immune surveillance. In contrast, specialized tissue-resident NK cells (trNK) and their close cousins, ILC1s, develop in various tissues and are retained there to carry out distinct functions. In the uterus, trNK cells and ILC1s are physiologically designed to support and promote pregnancy by promoting placental tissue invasion, immune suppression, and angiogenesis. We hypothesize that in the setting of EC the normal processes of uterine trNK cell and ILC1 development and function are co-opted by the tumor cells to promote their growth and invasion. Our goals in this proposal are to gain a comprehensive understanding of the cellular and molecular components that regulate human NK cell development in healthy tissues and to determine how these processes are impacted in the setting of EC. Our two specific aims are: 1) To define NK cell and ILC1 developmental pathways in human tissues; and 2) To determine how NK cell development and function are shaped by human EC. In particular, in Aim 1 we will test the hypothesis that all NK cells and ILC1s stem from a common group 1 ILC precursor cell, which we have recently identified in human lymphoid tissues. We propose a series of experiments to test our hypothesis and to determine the molecular regulation of the NK cell versus ILC1 developmental axis stemming from the novel precursor cell. Further, we will elucidate the developmental pathways of NK cells and ILC1s in the healthy human uterus, testing the hypothesis that uterine NK cells and ILC1s also stem from a similar group 1 ILC precursor cell but ultimately terminally differentiate through pathways distinct from those in lymphoid tissues. In Aim 2 we will test the hypothesis that NK cell development from the common group 1 ILC precursor is skewed towards the production of ILC1s and poorly cytotoxic uterine trNK cells that are permissive if not promoting of tumor growth. Through our proposed studies we will determine how NK cell development, functional diversity, and plasticity are shaped by EC. The clinical importance of these studies lies in the fact that EC is the most common gynecologic malignancy in the United States and is the sixth leading cause of cancer death in women. Further, we predict that the knowledge gained from our studies will improve our fundamental understand of human NK cell developmental biology and ultimately the design of future immune-based cancer therapies for EC and potentially other forms of cancer in which innate immunity is impacted.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Critical electrogenic proteins responsible for maintaining cardiac excitability and conduction, including sodium channels (NaV1.5), inward-rectifying potassium channels (Kir2.1), L-type calcium channels (Cav1.2), sodium-potassium ATPase (NKA), and sodium-calcium exchanger (NCX) have been identified to reside in distinct ion channel ‘pools,’ with localization at the cell-cell junction, the intercalated disk (ID). These distinct ion channel pools suggest regulation via both ‘global’ and ‘local’ control mechanisms. Within the ID, heterogeneous nanoscale structure results in channels concentrating around gap junctions and mechanical junctions, forming specialized nanodomains. ID nanodomains perturbation can induce proarrhythmic conduction defects, and disruption of these nanodomains has been identified in human arrhythmia patients, suggesting that these sites are key determinants of conduction. However, ID nanoscale structure and molecular organization and their implications for functional electrophysiology have yet to be systematically investigated in health or disease. In this project, we will undertake the first-ever comprehensive and granular quantification of ID structure and molecular organization using cutting-edge light and electron microscopy techniques and computational analysis. Further, we will develop a novel computational modeling framework to incorporate experimental measurements of these distinct ion channel pools (lateral membrane and ID) and ID nanoscale structure to assess regulation of tissue-scale cardiac conduction, for direct comparison with optical mapping of murine myocardium. Simulations will extend predictions to conduction in human ventricles and predict how both chronic and acute ID perturbations impact conduction in conjunction with additional functional defects, including non-ischemic heart failure. Upon successful completion of these aims, we will produce a new theoretical underpinning for which distinct ion channel pools and intercalated disk nanoscale structure confer a ‘global/local control’ of cardiac conduction and suggest new therapeutic approaches to preserve conduction during disease progression.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Immune checkpoint blockade (ICB) has produced extraordinary clinical responses in more than 25 tumor types. However, only a small number of patients benefit from this therapy owing to the immunosuppressive tu- mor microenvironment (TME). As one of the major components of the TME, tumor-associated macrophages (TAMs) usually possess profound inhibitory activity against tumor-killing T cells and facilitate tumor escape from immunotherapy. Clinical findings have shown that the presence of suppressive pro-tumorigenic TAMs correlates with reduced survival in bladder cancer patients treated with immunotherapy. Due to the plasticity of macrophages, excitement has been growing to reshape the pro-tumorigenic TAMs toward the anti-tumorigenic phenotype to stimulate the immunity against cancer. Emerging evidence reveals that this process of macro- phage polarization is inextricably affected by metabolites in the TME, such as free fatty acid (FFA). Lack of data regarding the role of FFA signals in macrophages prevents us from designing an elegant approach to re- polarizing TAM to foster a better anti-tumor T cell response. We propose to dissect the molecular basis of the FFA-mediated signal pathway in TAMs differentiation and its role in resistance to PD-1 blockade, which can be leveraged to restore sensitivity to ICB therapy. Our analysis of published datasets revealed that TAMs from human bladder tumors uniquely express elevated levels of a fatty acid receptor, G Protein-Coupled Receptor 84 (GPR84). These GPR84 expressing cells exhibited enriched hallmarks of anti-tumorigenic function com- pared to their counterpart. Further analysis reveals that expression of GPR84 significantly correlates with longer survival in bladder cancer patients. Our work shows that genetic ablation of GPR84 leads to enhanced production of inhibitory molecules including Arginase 1 via activation of CCAAT/enhancer-binding protein beta (C/EBPβ). By contrast, GPR84 activation by its agonist 6-OAU can reprogram pro-tumorigenic macrophages to produce anti-tumorigenic signature molecules, such as tumor necrosis factor-α (TNFα). Mechanistically, we found that GPR84 potentiates the activity of Nuclear factor kappa B (NF-κB) to enhance TNFα production. Central hypothesis: GPR84 serves as a metabolic signaling checkpoint for determining the function of macro- phage by restricting the immunosuppressive while promoting the immune-stimulating phenotype. Treatment with GPR84 agonists significantly retards tumor growth and increases the anti-tumor efficacy of anti-PD-1 mAb therapy in a MB49 bladder cancer model. Aim 1: Determine whether the lack of GPR84 promotes the polariza- tion of immunosuppressive TAMs. Aim 2: Dissect the molecular and epigenetic mechanisms by which GPR84 signaling promotes an immune-stimulating phenotype in macrophages. Aim 3: Determine whether targeting GPR84-mediated macrophage repolarization enhances the anti-tumor efficacy of ICB. Results will inform the development of promising treatments to reshape immunosuppressive TME through manipulation of metabolic signaling, and thereby restore responsiveness to PD-1 blockade for bladder cancer patients.

NIH Research Projects · FY 2026 · 2023-04

SUMMARY This project aims to mechanistically understand and combat pathological mechanisms activated by SARS- CoV-2 in the lungs and heart. We will further investigate whether these mechanisms are altered in specific immunodeficiencies linked to severe COVID-19. Our work will leverage major discoveries made by our group, including that the non-canonical inflammasome protein Caspase-11 (CASP11, homologous to CASP4 in humans) promotes pathological inflammation in SARS-CoV-2 infection, and that CASP11 KO mice experience significantly less severe infections than than WT mice. In Aim 1, we will test the hypothesis that cell-specific roles of CASP11 mediate SARS-CoV-2 pathogenesis by examining infections of cell-specific CASP11 KO mice (CASP11 flox allele mice). Further, we will target CASP11 downstream effectors identified by our work, such as the chemokine CXCL1 and recruited neutrophils, to determine their roles in unique aspects of SARS-CoV-2 pathogenesis. We will examine additional emergent candidate CASP11-dependent molecules, using genetic, neutralizing antibody, and chemical inhibitor strategies. This aim will yield new understanding of roles for CASP11 in specific cell types and will identify tailored strategies for preventing or treating unique aspects of COVID-19 pathology. In Aim 2, we will test the hypothesis that CASP11-dependent mechanisms of pathogenesis are exacerbated in specific immunodeficiencies linked to severe COVID-19. These include type I interferon (IFN) and IFN-induced transmembrane protein 3 (IFITM3) deficiencies. We have shown IFN alpha receptor KO and IFITM3 KO mice model these deficiencies, including exacerbated lung infections and virus dissemination to the heart. The use of these models in combination with CASP11 KOs will allow identification of pathogenic mechanisms in the lungs, and in the hearts of mice with or without direct cardiac tissue infection. Overall, our work will reveal fundamental mechanisms of SARS-CoV-2 pathogenesis in the lungs and heart, including involvement of specific cell types, pathways, and molecules, thus revealing targetable therapeutic strategies for combating COVID-19 in immunocompetent, as well as highly vulnerable, populations.

NIH Research Projects · FY 2026 · 2023-04

Atrial fibrillation (AF) is the most common arrhythmia and has a high risk of mortality and morbidities. Among the general AF population, new-onset postoperative atrial fibrillation (POAF) is the most common complication after open- heart surgery, which significantly increased mortality and amplifies hospital and patient costs. The mechanisms underlying POAF are unclear, thus effective prediction and/or prevention remain unavailable. This proposal aims to fill this knowledge gap by identifying stress-response kinase JNK as a novel POAF biomarker for surgery patients and exploring the translational potential of local JNK inhibition in atria as a novel anti-POAF therapeutic approach. Predisposing factors for POAF include advanced age, binge alcohol, as well as intraoperative and postoperative atrial injury and/or ischemia. One common element among these factors is tremendously increased cellular stress, which is known to activate the c-Jun N-terminal kinases (JNKs), an important stress-response kinase. We recently discovered and reported a previously unrecognized causal link between cardiac JNK activation and abnormal cell-cell communication (via gap junction channels) as well as abnormal Ca triggered activities which enhance AF propensity. Our intriguing preliminary findings suggest that atrial JNK activation is well correlated to POAF incidence in patients within 10 days of coronary artery bypass graft (CABG) surgery, indicating POAF likely involves JNK activation and possible JNK-driven atrial arrhythmogenesis. Intriguingly, our preliminary results show for the first time that JNK is present in blood. And JNK activation in the heart increases blood JNK. Accordingly, the concordant atrial JNK activation and rise in plasma JNK levels correlates nicely to the increased incidence of AF. Next, we found that most of the plasma JNKs are carried by microparticles (MPs) circulating in the blood. Our pilot data further suggest that heart cells shed JNK-microparticles (JNK-MPs). All these intriguing preliminary results combined with our previous findings point to a unique heart-blood JNK relationship that links to AF pathogenesis. Here, we will use a series of cutting-edge biochemical assays and electrophysiological techniques on surgically removed intact atrial tissue, isolated atrial myocytes and blood samples from CABG patients and human donor hearts as well as animal models recapitulating AF risk factors (aging & binge alcohol). Our Specific Aims are: 1) Establish the electrophysiology & biochemistry profiles of JNK, pro-inflammatory cytokins, and arrhythmic substrates in atrial tissue/blood of CABG patients and their correlation to POAF incidence; 2) Prove activated JNK in the blood is a biomarker of POAF risk and test a potential AF therapeutic atrial painting gene transfer intervention in aged rabbits. Establishing the atrial/blood JNK as a possible biomarker of POAF risk is entirely novel here. Clinical accessibility for both atrial tissue and blood samples and POAF events make CABG patients an ideal population for studying the JNK-AF link in humans. Developing an atrial painting gene transfer intervention that could be applied to high POAF risk patients during surgery is innovative. The unique combination of cardiac research expertise and novel electrophysiology/biochemistry techniques makes this proposal technically & experimentally innovative.

NIH Research Projects · FY 2025 · 2023-04

This study proposes longitudinal research among older adults to assess the associations of uncontrolled pain, co-occurring chronic conditions or geriatric symptoms (multimorbidity), and opioid–drug interactions with risk for opioid use disorder (OUD) or overdose (OD). This study is responsive to AHRQ’s special interest (NOT-HS- 21-010) in research to address substance use disorders, including OUD and OD in older adults. Older adults have experienced significant increases in OUD or OD in the last decade despite a decrease in the number of opioid prescriptions, signaling an urgent need to identify factors beyond prescription opioid use that contribute to OUD/OD to inform interventions in older populations. Pain, multimorbidity, and polypharmacy, all strongly associated with advancing age, are frequently noted as potential risk factors for OUD/OD in older adults, but relevant evidence is lacking. Relieving pain is the most frequent motive for opioid misuse in older adults, but to what extent uncontrolled pain contributes to risk for OUD/OD is unknown. Multimorbidity is highly prevalent in older adults with OUD or OD, but its association with uncontrolled pain and the joint association of multimorbidity and uncontrolled pain with risk for OUD/OD are unknown. There is also limited understanding regarding the risk of OUD/OD when prescription opioids for pain treatment are concurrently used with drugs for other chronic comorbid conditions that may interact with opioids in older adults. To fill these research gaps, this proposed study will leverage a 100% Medicare sample and Medicare data linked to nationally representative longitudinal survey data to study the associations of pain, multimorbidity, and opioid-drug interactions with risk for OUD/OD outcomes of older adults with chronic pain. Specifically, we will examine the associations between pain control and risk for OUD or OD in older adults diagnosed with chronic pain and treated with prescription opioids. We will also evaluate the association of multimorbidity with pain control and their joint association with OUD/OD risk in older patients. Furthermore, we will assess the extent to which concurrent use of opioids with drugs for treating common comorbid conditions with chronic pain in older adults is associated with OUD/OD outcomes in older adults with chronic pain. We will measure both pain and high-impact pain and quantify multimorbidity through both a diagnosis-based index and the presence of geriatric symptoms that are underdiagnosed or not diagnosable. Our study will provide empirical evidence on the extent to which uncontrolled pain, multimorbidity, and opioid-drug interactions may play a role in increasing risk for OUD or OD among older adults with chronic pain. Given the accelerated growth of the US aging population and the continuing increase in OUD, OD, and opioid misuse, our data will inform interventions that integrate care for chronic pain and other chronic comorbid conditions to improve pain control while reducing OUD or OD in older adults.

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT Tuberculosis (TB), caused by Mycobacterium tuberculosis (M.tb), is a leading infectious disease and cause of death worldwide. The growing burden of drug-resistant (DR)-TB is complicating TB treatment. Early diagnosis of TB with drug susceptibility testing (DST) is critical for successful treatment and is the first pillar of the World Health Organization’s (WHO) End TB Strategy. DST is achieved via phenotypic or genotypic methods. Traditionally, phenotypic DST is performed on solid (Löwenstein Jensen) or liquid media (MGIT) in a two-step process: first a culture to identify M.tb growth, and then re-culture of the isolate with the drugs to be tested. In addition to requiring biosafety level II-plus labs, the DST process, if available in low-middle income settings, can take 42 to ~6 months from sample collection to notification of results to the clinical provider resulting in treatment delays, continued transmission, and higher mortality. Conversely, genotypic DST has many advantages, including a reduced time to result (< 2h for GeneXpert) and the possibility of deployment to at or near point of care (POC). However, its widespread use in high TB burden resource-limited settings is hindered by the need for regular power supply and importantly cost. Thus, NIH/NIAID is redirecting attention to innovative and simple phenotypic DST solutions to be deployed at or near POC. The goal of this application is to develop the 1G test into the 2G test, providing higher flexibility to perform DST for 1st and 2nd frontline drugs, including drugs prescribed for DS- and DR-TB regimens such RIPE (DS-TB oral regimen composed of RIF/INH/PZA/Ethambutol), HPMZ (DS-TB 4-month short course oral drug regimen composed of INH/Rifapentine/MFX/PZA) and BPaL [MDR- and pre-XDR oral drug regimen composed of bedaquiline (BDQ), pretomanid (PMD) and linezolid (LNZ)], as well as clofazimine (CFZ) and delamanid (DLM), other WHO recommended oral agents for DR-TB. Because the 2G test is non-proprietary, its cost is expected to be extremely low (< $8) and mainly driven by the cost of drugs. Further, for the 1G test we tested a simple step to digest/decontaminate sputa that does not require equipment, meeting the near to POC test definition. We will optimize this sputum-processing protocol for use with the 2G test. We propose to: Aim 1) Develop and validate the 2G test by defining the stability and critical concentration (CC) for new drugs against known DR-M.tb strains, and optimize appropriate sputum digestion and decontamination protocols for this test; Aim 2) Determine the agreement of the 2G test with current gold standard methods for phenotypic DST for each of the 11 drugs, and Aim 3) Determine the accuracy of the 2G test against reference phenotypic DST protocols using freshly collected sputa in field settings and assess its usability, acceptability, and feasibility. We expect that the novel, simple, affordable and sustainable 2G test will provide a significant improvement when compared to current phenotypic DST reference methods, allowing rapid and tailored treatment for DS-/DR-TB in low- and middle-income countries with high TB burden.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/Abstract Acute pancreatitis (AP) accounts for over 300,000 admissions in the U.S. with annual costs exceeding $3 billion. Most cases of AP are mild (MAP) with a hospital stay of 3-4 days, but approximately 15% of AP subjects develop severe disease (SAP), defined by presence of persistent organ failure. Up to a third of SAP patients expire from multi-system organ failure after weeks in the intensive care unit. To date, no therapeutic agents have been successful at ameliorating the protracted hospital course of SAP. The National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) workshops identified two critical knowledge gaps as barriers to developing pharmacologic interventions: 1) the establishment of a highly accurate, early prediction tool to identify which subjects will develop SAP during hospitalization; 2) a more in-depth knowledge of SAP mechanistic pathways and immuno-pathogenesis to identify novel therapeutic targets. We propose the MoSAIC Study (iMmune SIgnAtures and ClIniCal outcomes in AP), a prospective multi-center, observational cohort that will address both of these knowledge gaps in SAP. We recently discovered a novel multi-cytokine panel (comprising of angiopoetin-2, hepatocyte growth factor, interleukin-8, resistin, and tumor necrosis factor-α receptor-1) that accurately predicts SAP early in the disease process with an accuracy of 0.89 and significantly outperforms existing prediction tools. Hence, the first aim of this project is to validate this multi-cytokine panel for SAP in large AP cohorts consistent with NIH inclusion policy across multiple U.S. clinical sites (Specific Aim 1). In preliminary studies, immunologists at the Benaroya Research Institute (BRI) have unique immune cell changes such as an increase in monocytes and a decrease of T follicular helper and memory B cells in blood samples of AP patients compared to healthy controls. The MoSAIC study will extend this work by defining the circulating immune cells that correspond with cytokine signatures in early AP and identifying the immune pathways driving the development of SAP (Specific Aim 2). This will generate the first high-dimensional phenotypic analysis of immune cell types in human AP and provide new insights into its immune mechanisms. MoSAIC investigators have NIH-funded complementary expertise in pancreatitis, immunology, and bioinformatics. The team is led by well-published pancreatologists at the Ohio State University and immunologists at BRI, supported by a dedicated bioinformatics core at BRI. It also includes three additional academic medical centers with proven track-records of enrolling population cohorts consistent with NIH inclusion policy. This proposal is an approved ancillary study of the NIH/NIDDK Type 1 Diabetes in Acute Pancreatitis Consortium (T1DAPC), of which all MoSAIC sites are members. Successful completion of the MoSAIC study will have the following impact by: 1) providing groundbreaking insight into the early immune events of SAP based on robust human data, 2) identifying therapeutic immune targets for further testing, and 3) establishing a U.S. multicenter research platform for launching clinical trials to test immunotherapies in AP.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY Recent advance in the prevention and treatment of breast cancer significantly increase the survival of breast cancer patients. It will be highlighted that with the increased survival of breast cancer patients, however, the incidence of latent breast cancer brain metastasis (BCBM) significantly increased. BCBM is one of critical factors that contributes to the lethality of breast cancer patients. Autophagy is important in the initiation and progression of cancers, neurodegenerative diseases, metabolic disorders, and infectious diseases. Nevertheless, the functions of autophagy in tumor microenvironment, especially the roles of glial autophagy in BCBM, are not investigated. Using syngeneic mice, we established intracerebral and intracarotid artery injection models for BCBM with different mouse breast cancer cells. We observed robust astrogliosis, infiltration of microglia, and uncontrolled growth of malignant cells in mouse BCBM models. More importantly, we found increased autophagy in reactive astrocytes but not in neurons at the borders of brain-breast cancers. Using brain specific Fip200 (an autophagy essential gene) conditional knock out mice, we revealed the indispensable functions of autophagy in astrocytes for the initiation and progression of BCBM. Breast cancer cells activated Stat3 to initiate brain metastasis, the process of which depended on intact autophagy in tumor microenvironment astrocytes. Moreover, we confirmed the elevation of autophagy in astrocytes and activation of Stat3 in cancer cells in dissected human samples, suggesting a correlation of our experimental observation with clinical importance. Our results indicated a previously unrecognized mechanism for astrocytes in tumor microenvironment to support the initiation and growth of breast cancer cells in brain. To facilitate mechanistic studies, we generated a coculture model using human breast cancer cell lines with human embryonic stem cells derived cerebral organoids. Our data indicated that the metastatic human MDA-MB-231 breast cancer cells could colonize in cerebral organoids while the non- metastatic MCF-7 cells could not grow. In this proposal, we will determine the molecular mechanisms by which autophagy in astrocytes is utilized by breast cancer for brain metastasis, using a combination of molecular biology, cell tracing method, brain injections, stem cell technology, and mouse genetic approaches. Our proposed studies will have significant impact on understanding the fundamental mechanisms of reactive astrocytes to regulate BCBMs. Our research will raise new translational approach for stratification and treatment of BCBM patients. The results in our research might also shed light on the investigation of tumor microenvironment for other brain metastatic cancers and primary brain tumors.