Ohio State University

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Psychotic disorders are serious illnesses that lead to significant functional impairment, even during the early course of illness (i.e., first-episode psychosis; FEP). Social cognition is a strong predictor of key functional outcomes in psychosis (i.e., social and occupational functioning) and among social cognitive domains, mentalizing – the ability to infer and reason about the mental states of other people – is most robustly impaired and simultaneously most predictive of outcomes. However, existing interventions addressing social cognitive dysfunction in psychosis are minimally effective in improving real-world outcomes. Further, though psychosis studies have revealed abnormal function in brain areas comprising the mentalizing network (MN; e.g., medial prefrontal cortex and temporoparietal junction), the vast majority of existing studies have utilized fMRI paradigms that use non-human or non-interactive stimuli that may limit ecological validity and further fail to examine associations between neural function and real-world outcomes. Collectively, social cognition is a critical factor in psychosis that remains poorly understood. Thus, the proposed study will combine neural, behavioral, and real- world functioning measures to delineate brain-behavior mechanisms of social cognition in FEP. The study will enroll 45 FEP individuals and 45 matched non-clinical controls (NCC) and assess their neural (i.e., fMRI) and behavioral (i.e., validated laboratory-based measures) social cognitive function as well as their real-world social and occupational behavior. Data from this study will allow for an innovative investigation of whether MN dysfunction is associated with social cognitive performance on validated behavioral tasks both across and within FEP and NCC participants (Aim 1), and how these brain-behavior measures of social cognitive performance relate to laboratory-based measures of social and occupational functioning (Aim 2). The current study will also examine the extent to which MN function predicts real-world social and occupational functioning assessed via ecological momentary assessment (Aim 3). This proposal concurrently provides an excellent opportunity to receive mentor-directed, hands-on training in several key areas necessary to refine the candidate’s knowledge and skill-sets. Though the candidate has extensive clinical research experience in FEP and has completed foundational fMRI training, she proposes a training plan focused on three new domains: 1) advanced social- cognitive neuroscience and fMRI task development, 2), linking of fMRI to behavioral constructs of social cognition and functional outcome, and 3) translation of fMRI and behavioral constructs to psychosis. The proposed study coupled with completion of the training plan will effectively launch the candidate toward her long-term goal of an independent career in the translational neuroscience of social-cognitive dysfunction in early psychosis, and will lay the foundation for future, high-impact R01 studies focused on refined identification and targeting of biobehavioral factors influencing social and functional outcomes.

NIH Research Projects · FY 2026 · 2023-08

PROJECT SUMMARY/ABSTRACT Human-associated microbes are causally linked to processes as diverse as immunomodulation, protection against pathogens, and atherosclerosis. Many of these links are mediated through biochemical transformations of dietary, drug, or host compounds. This makes the microbiome a promising therapeutic target, especially as we could potentially affect downstream processes by controlling metabolic inputs. However, in order to effectively intervene, we must first understand how exactly changes in these inputs lead to differential regulation of growth, gene expression, and metabolism. This is challenging because our microbiomes are not only genetically and physiologically diverse, but are also highly diverged from the most common model organisms, with many genes of unknown function. Over the next five years, my research group will use a combined computational and experimental strategy to characterize gene function, metabolic regulation, and microbial interactions in one of the most prevalent and abundant clades of gut bacteria, the Bacteroidales. Specifically, we seek to determine 1) which Bacteroidales genes are involved in growth on different nutrients and stressors; 2) how Bacteroidales genes are regulated, and how this affects their metabolic outputs; and 3) how Bacteroidales interact with the other microbial inhabitants of the gut. We will accomplish this by gathering high-throughput in vitro data from diverse sets of microbiome isolates and synthetic communities, developing more powerful and specific statistical tools to analyze these data, and using these new data and tools to re-analyze metagenomics data gathered from in vivo case-control studies. I envision that this line of inquiry will provide missing fundamental knowledge about this clade of microbes, which will ultimately help us interpret case-control studies of the microbiome and support the development of more precise interventions.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Quantitative literacy is an essential component of rigorous research. Quantitative literacy incorporates mathematical knowledge, statistics, reasoning, and data and visual literacies to assign meaning to numerical data. In scientific research, being quantitatively literate includes being proficient in contextualizing quantitative datasets and reading and reporting numerical data. Being quantitatively literate is critical to identifying and avoiding weakness in rigor. Our team will design engaging, clear content for the purposes of providing a foundational understanding what quantitative literacy is and its role in scientific rigor, developing strategies and practices in interpreting, analyzing and contextualizing numerical data analysis, and educating learners on best practices in reporting numerical data to promote scientific rigor. Our educational design invites novice, intermediate and advanced learners to review and interact with quantitative literacy concepts through five different learning units. Users have the option to progress through the units covering foundational principles of quantitative literacy, practices in contextualizing data, interpreting statistics, evaluating and utilizing illustrative reports and reporting quantitative data, or can acquire knowledge through independent activities, catering to individualize learning needs. Our expertise and practice in quantitative literacy as well as in building educational programs will be utilized to construct educational experiences that will ultimately improve scientific rigor.

NIH Research Projects · FY 2026 · 2023-08

High risk for premature mortality is one of the most pressing issues faced by the growing population of aging autistic adults. Autistic adults are disproportionately more likely to have chronic conditions, leading to increased risk for mortality compared to the general population. However, one major barrier to identifying those at greatest risk for mortality is the absence of accurate predictive tools for this population. Our objective is to establish a novel, machine-learning derived mortality risk index for autistic older adults. We will leverage our team’s unique expertise in autism aging research, population-level administrative data analysis, and machine learning to achieve our specific aims: (Aim 1) identify comorbidities and geriatric complaints that differentially influence time- to-mortality for autistic and non-autistic older adults; (Aim 2) compare existing mortality risk indices to a novel, autism-specific index for predicting autistic older adults’ risk of mortality; (Exploratory Aim 3) determine the distribution of mortality risk among autistic older adults in local healthcare systems as a precursor for prospective studies. We will achieve these aims through the synergistic use of national administrative billing data and local electronic health records data. In Aim 1, we will apply a machine learning technique called “logic forest” in an innovative way to identify specific comorbidities and age-related conditions, or combinations thereof, that differentially influence time-to-mortality among autistic and non-autistic older adults using the most recent nine years of national Medicare data. In Aim 2, we will apply a stochastic hill climbing optimization technique, a type of machine learning, to national Medicare data to develop an algorithm-based index that quantifies autistic older adults’ risk of mortality based on comorbidities and demographic characteristics. We will compare the predictive validity of our novel autism-specific algorithm-based index to the Charlson and Elixhauser comorbidity indices, the gold-standards of mortality risk measurement among the general population. Last, in Exploratory Aim 3 we will obtain sample size estimates for prospective studies by quantifying mortality risk among aging autistic adults in two large healthcare systems using the Charlson, Elixhauser, and our novel autism-specific mortality risk indices. Findings of this study will have practical applications for researchers to identify participants for prospective observational and intervention studies and clinicians to identify high-risk cases for special management and intervention. This study is responsive to NOT-AG-21-020 in that we will analyze existing Medicare claims data to examine “subgroups of older adults with special needs” and “health outcomes in complex multimorbid older adults”. Further, this study is aligned with the NIA’s Strategic Plan as we seek to “understand disparities related to aging and [inform] strategies to improve the health status of older adults” on the autism spectrum. This project will have a high public health impact yielding critical new information about mortality risk among a historically understudied aging population that can ultimately be used to improve life-expectancy among autistic people.

NIH Research Projects · FY 2025 · 2023-08

Project Summary/Abstract The Multidisciplinary Research Training in Dental, Oral, and Craniofacial Sciences (MARTDOCS) program is built on the strong foundation of more than 20 years of successful training of PhD, DDS-PhD, and postdoctoral scholars by the Ohio State University (OSU) College of Dentistry (COD) Oral Biology Graduate Program (OBGP) and its partner programs in the OSU College of Medicine, the Biomedical Science Graduate Program (BSGP) and Neuroscience Graduate Program (NSGP). The overarching goal of this integrated pre-doctoral and postdoctoral training program is to foster the development of future leaders in dental, oral, and craniofacial research who are specifically equipped to employ multidisciplinary research strategies in basic, translational, and clinical science, a high-priority objective for the NIDCR. We will achieve this goal by harnessing the enormous institutional resources available at The Ohio State University and assembling a collaborative team of productive and dedicated mentors who bring complementary expertise to the program. Training opportunities include: 1) pre-doctoral training in the OBGP PhD program in the OSU COD and two inter-disciplinary graduate programs, the BGSP and NSGP, in the OSU College of Medicine; 2) dual degree training for the DDS/PhD degrees (DSTP); and 3) postdoctoral training including post PhD and clinician/scientist programs. Nationally recruited trainees will gain invaluable training in research methodology and career development through structured programs and courses on cross-cutting themes ranging from molecular and cell biology, inflammation and immunity, big data informatics, and clinical oral health research. In addition to the primary mentor and a mentoring team, each trainee’s progress will be monitored by an executive committee, and the graduate studies committee for each graduate program. The program will capitalize on innovative practices at the OSU COD for recruitment and retention of under-represented minorities and individuals with disabilities. Over the past 10 years, 36 trainees have matriculated into the program and have been mentored by 14 different faculty members. Our proud training history confirms high level mentoring and support for trainees throughout their entire career pipeline, including during training (e.g. competitive research awards at IADR, AADOCR and other conferences; F30/31 grants awarded), at critical transition points (e.g. postdoctoral, residency, and faculty positions attained), and in long-term benchmarks (e.g. promotion; independent funding; and international recognition of trainee achievements). In summary, this training program offers an innovative interdisciplinary science curriculum based on the mechanisms of human disease, and an extensive and flexible curriculum in oral sciences, coupled with the long-term mentorship of experienced interdisciplinary faculty. As a top 20 NIH-funded state dental school that is housed in a university with over 40 state-of-the-art core research facilities and 15 interdisciplinary research centers, the MARTDOCS at OSU is extremely well-equipped to produce highly trained scientists and clinician- scientists who can lead the field of oral health research into 2030 and beyond.

NIH Research Projects · FY 2025 · 2023-07

SUMMARY Triple-negative breast cancers (TNBCs) are highly aggressive and often relapse post standard cytotoxic chemotherapies. The immunotherapies such as immune checkpoint blockers (ICBs) that target PD-1, PD-L1 or CTLA-4 represent a major breakthrough in cancer treatment, but 75-90% of TNBC patients failed to respond due to primary and acquired resistance. The Sacituzumab-SN38 and Pembrolizumab-chemotherapy have been developed and applied to treat refractory metastatic TNBC, showing the great potential of combined and targeted therapies, but novel effective treatment strategies for TNBC are urgently needed. We recently detected transmembrane CD276 (B7-H3) associated with angiogenesis, metastasis and immune tolerance in most TNBC patients, and developed a humanized anti-CD276 monoclonal antibody (mAb) capable of targeting CD276+ TNBC and upregulating tumoral immunity. Furthermore, we established innovative platforms for concurrent conjugation of highly cytotoxic emtansine and immunoregulating toll-like receptor (TLR) agonist in one antibody- drug conjugate (ADC). Our preliminary evaluations showed that the CD276-targeted dual-payload ADC (276- DualADC) effectively killed multiple TNBC subtypes, significantly enhanced immune functions and overcame ICB resistance to PD-1 mAb, and reduced tumor burden by 90-100% and metastasis in three animal models. These results indicate 276-DualADC is a promising therapeutic to treat TNBCs. Our goal is to develop and examine the effectiveness of combining humanized CD276 mAb-directed Hu276-DualADC, which targeting delivers a potent chemotherapy and immunotherapy TLR 7/8 agonist, and PD-1-targeting ADC to eliminate heterogenous and metastatic TNBC cells in vivo. It is hypothesized that this novel combinatory strategy, named as Hu276/PD-1-DualADC, synergizes multiple chemo- and immuno-mediated anti-cancer mechanisms, i.e., direct cancer cell killing, tumoral immunity, tumoral cytokine, and immune checkpoint blockade, to enhance TNBC treatment efficacy. Three aims were proposed to test the hypothesis. Aim 1 will produce large-scale Hu276-DualADC carrying mertansine and imidazoquinoline, and characterize its affinity, TNBC-specificity, biodistribution and toxicity. Optimal treatment strategy will be determined in maximal tolerated dose and pharmacokinetics studies. Finally, anti-TNBC efficacy will be evaluated in primary xenograft models and distant metastatic models. Aim 2 will assess the synergistic effects of HuCD276/PD-1-DualADC in three immunocompetent models. The underlying mechanisms (proliferation, apoptosis, immune cell infiltration and activation, tumoral cytokine, and ICB restoration) will also be delineated. Aim 3 will fully evaluate the long-term therapeutic efficacy in metastatic syngeneic TNBC xenograft models post surgical resection and patient-derived xenograft (PDX) models. Pre-IND toxicology will also be investigated to collect preclinical data for future clinical trial launching. Successful completion of this project will provide a new strategy to treat aggressive TNBCs.

NIH Research Projects · FY 2025 · 2023-07

Project summary CD19 targeted chimeric antigen receptor T cells (CAR-Ts) have revolutionized cellular immunotherapy in refractory B-cell malignancies such as B-cell Non-Hodgkin’s lymphoma (B-NHL) and B-cell acute lymphoblastic leukemia (B-ALL). Despite high rates of complete remissions, relapses, 50% of which within the first year, remain a critical challenge highlighting the need for novel CAR-T cell products. Two patterns of relapse are observed: (i) Antigen-negative relapses, caused by target antigen loss and (ii) antigen-positive relapses, which are mediated by lack of persistence or loss of function of the CAR-Ts. Commercial CD19 CAR-Ts employ either a CD28 or a 4-1BB costimulatory domain however, a novel OX-40 domain has been shown to promote persistence, cytotoxicity and decrease exhaustion. Finally, increasing evidence suggests that stem-like memory T cell phenotype in the CAR-T product is associated with more durable responses. Schneider et. al developed a Trispecific CD19, CD20, CD22-targeting CAR-Ts with an OX-40 costimulatory domain and showed significant activity in preclinical lymphoma models compared to CD19 CAR-Ts. We validated these findings with in-house manufactured Trispecific CAR-Ts and confirmed their specificity, cytotoxicity, and immunophenotypic fitness in preclinical B-cell lymphoma models. Our proposal seeks to address the limitations of commercial CD19 CAR-Ts through a first-in-human in-house manufactured Trispecific CAR-Ts with an OX- 40 costimulatory domain for relapsed, refractory B-cell malignancies. These trispecific CAR-Ts are manufactured in the Ohio State University Cell therapy laboratory using the CliniMACS Prodigy device over a 6-day manufacturing process which allows for infusion of stem-like memory CAR-Ts. We hypothesize that trispecific CARTs will (i) reduce the risk of relapse mediated by antigen negative clonal escape; (ii) enhance persistence of CAR-Ts which will translate into deeper and more durable clinical responses. To address this, we propose two Aims. In Aim 1, we aim to conduct a first-in-human phase I trial with in-house manufactured trispecific CAR-Ts in patients with relapsed/refractory B-NHL, B-ALL, B-prolymphocytic leukemia, and chronic lymphocytic leukemia. Primary endpoints are feasibility, safety of trispecific CAR-Ts along with establishing a recommended phase II dose. Secondary endpoints are efficacy and duration of response. In Aim 2, we aim to identify key mechanisms of efficacy and resistance to trispecific CAR-Ts. Specifically, we plan to assess: (i) persistence of CAR-Ts and its correlation with complete remission rates, duration of response as well as incidence and severity of adverse events; (ii) immunophenotypic and transcriptional features of impaired function of CAR-Ts and other mononuclear cells using state of the art high-dimensional spectral flow cytometry and CITE-sequencing. At completion of this project, our work will have established feasibility of in-house manufacturing of trispecific CAR-Ts and provided insights into mechanisms of relapse and efficacy.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY Mantle cell lymphoma (MCL) is an incurable non-Hodgkin lymphoma, and despite intensive therapeutic approaches, the median progression-free survival after first-line treatment is four years. The emergence of chemoresistance is rapid, durable responses to second and third-line therapies are rare, and relapse is virtually universal in MCL. Cell cycle dysregulation, primarily by upregulation of E2F1 target genes, is a hallmark of MCL. We identified a novel role of Fibroblast growth factor receptor-1 (FGFR1) in MCL survival and regulating cell cycle-dependent processes primarily by inhibiting E2F1 mediated transactivation. We show that FGFR1 and not other homologs are overexpressed in MCL patients, and its expression is associated with a poor prognosis. Functionally, genetic ablation of FGFR1 or pharmacological targeting with erdafitinib, a selective small molecule targeting FGFRs, induced cell cycle arrest, cell death in-vitro, reduced tumor formation, and improved overall survival in-vivo. Mechanistically, we show that FGFR1 positively regulates E2F1-mediated transactivation of its target gene through cMYC/EZH2/CDKN1C axis, contributing to cell survival. Hence, we hypothesize that the FGFR1 signaling pathway is a critical modulator of cell survival and represents a novel and attractive candidate for targeted therapy for patients with relapsed MCL. In this proposal, we will dissect the mechanisms by which FGFR1 impacts MCL survival and perform preclinical studies using erdafitinib as a therapy to prevent and treat MCL through the following specific aims: SA1: To characterize the role of FGFR1 in MCL survival in-vitro and in- vivo mouse models of MCL. In this aim, we will investigate a) FGFR1's role in regulating E2F1 mediated transactivation of target genes and survival in-vivo, b) the role of CDKN1C in regulating E2F1 dependent transactivation program, c) the role of E2F1 target gene CDK1, to positively regulates cMYC stability and constitute a feedback loop and d) identify mechanism-based combination strategies to maximize the therapeutic potential of FGFR1 inhibition. SA2:To conduct a preclinical investigation of inhibition of FGFR1 using patient- derived xenograft (PDX) murine models and a primary genetic murine model of MCL. We will conduct a preclinical trial of erdafitinib using PDX and murine models of MCL to identify a) the doses of erdafitinib that induce efficient inhibtion of FGFR1 signaling and downstream target gene expression without unacceptable toxicity and b) the best dose and schedule of erdafitinib resulting in maximum clinical efficacy alone and c) in combination with an ibrutinib. SA3: We will perform a retrospective analysis on tissue biopsies from relapsed MCL patients to a) analyze the effects of FGFR1 protein and its associated clinical outcomes and b) analyze the correlation between FGFR1 protein and, its activation, downstream effectors in patients with relapsed/refractory MCL. Impact: Our proposal will establish the role of FGFR1 in regulating E2F1 dependent transactivation program in MCL and provide a robust preclinical assessment by evaluating the toxicity, exposure, and efficacy of erdafitnib in preclinical models of MCL to translate FGFR1 inhibitor erdafitinib to treat MCL in clinics.

NIH Research Projects · FY 2025 · 2023-07

Abstract Microtubule-binding chemotherapeutics such as vincristine are among the most widely used anticancer agents in oncology for the treatment of multiple solid tumors and leukemias in children and adults. The clinical use of vincristine is associated with a debilitating, dose-limiting peripheral neurotoxicity for which no effective preventative treatments are presently available. In addition, the mechanism by which vincristine accumulates into dorsal root ganglion (DRG) neurons remains unclear to this day. Using a transporter screen of xenobiotic uptake carriers in heterologous overexpressed models, we recently found that the organic anion transporting polypeptide OATP1B3 (in mice, OATP1B2; collective referred to as OATP1B2/3) is an efficient transporter of vincristine that is expressed in human and mouse DRG neurons. Functional validation studies in OATP1B2- deficient mice and secondary screens confirmed that vincristine is transported into DRG neurons by OATP1B2. Furthermore, deficiency of OATP1B2 protected mice from vincristine-related changes in various hallmarks of peripheral neurotoxicity without altering the plasma levels of vincristine. To provide proof-of-principle and demonstrate translational relevance of this transport mechanism, we found that several known pharmacological inhibitors of OATP1B, including rifampin and the tyrosine kinase inhibitor, nilotinib, can preserve DRG neuronal function following treatment with vincristine without affecting its plasma levels or its cytotoxic potential against multiple acute leukemia cell lines. Finally, we identified α-tocopherol (vintamin E) as a previously unrecognized biomarker of neuronal OATP1B2/3 function that can be measured in the systemic circulation, and we validated the translational utility of this biomarker in a mouse model receiving treatment with OATP1B inhibitors. Based on these preliminary findings, we now outline three sets of related studies that will further test and refine the validity of our central hypothesis that targeted modulation of OATP1B2/3 function with optimized doses and schedules of novel OATP1B2/3 inhibitors can specifically affect accumulation of vincristine in DRG neurons and affect downstream toxic events without negatively influencing its plasma pharmacokinetic profile or anti-leukemic properties: (i) mechanistic characterization of nilotinib as the proof-of - principle OATP1B inhibitor, and identification and validation of additional modulators derived from a library screen that includes FDA-approved agents; (ii) biomarker-driven optimization using α-tocopherol as a companion diagnostic to guide dose selection of OATP1B modulators for in vivo testing; and (iii) safety and efficacy analyses of optimized combinatorial regimens of OATP1B inhibitors with vincristine, including simultaneous assessment of neuroprotection and anti-leukemic properties in established experimental models of acute leukemia. It is expected that these collective studies will not only shed light on the etiology of vincristine-induced peripheral neurotoxicity, but will be of translational relevance and provide a rationale for the future implementation of novel targeted intervention strategies to prevent this debilitating side effect.

NIH Research Projects · FY 2024 · 2023-07

1 Abstract 2 Rotaviruses (RVs) of group A (RVAs) remain an important cause of acute gastroenteritis and mortality in young animals 3 and children. Despite their significance, mechanisms of RVA cell entry, replication and attenuation remain poorly 4 understood indicating a critical need to address these knowledge gaps and design universal control approaches. 5 We have previously conducted comparative sequence analysis that identified two key amino acid substitutions 6 associated with cell culture adaptation and attenuation: D385N in the VP4 of most human and porcine RVA strains and 7 D393H in porcine OSU strain VP4 only. While these or similar mutations were previously suggested to be important for 8 cell culture adaptation/attenuation, their function has not been confirmed experimentally. Using our porcine intestinal 9 enteroid (PIE) system, we have demonstrated that contrasting modes of interactions with the host glycans determine 10 replication efficacy of different RVA strains. OSU replication was uniquely and significantly down-regulated by the 11 removal of terminal sialic acids, while the latter significantly enhanced the replication of a novel porcine G9P[13] strain. 12 OSU is a historic dominant RVA variant characterized by robust in vivo and in vitro replication and consistently 13 associated with diarrheal disease in piglets for several decades, while G9P[13] and G9P[19] are recent, globally 14 emerging RVA variants in swine and humans. It is possible that the latter strains have evolved to acquire some biological 15 advantage allowing for their efficient spread and replication in different hosts. Virulent OSU and G9P[13] strains have 16 unique aa substitutions in the aa positions 385 and 393 of the VP4 hydrophobic loop, which could be a reason for 17 variable interactions with host glycans. Additionally, our recent study demonstrated that the host (PIE) transcriptome 18 response associated with OSU and G9P[13] infections differed drastically altering glycan expression, cholesterol 19 metabolism and innate immune signaling in a strain-specific manner. These findings suggest that evolutionary 20 adaptation of RVAs to the host and host glycans is a critical mechanism allowing for their interspecies (including 21 zoonotic) transmission. 22 Following a previously optimized protocol, our lab has successfully cloned 11 segments of virulent RVA OSU into the 23 pT7 plasmid and rescued infectious virus. We now propose to use this reverse genetics system for in-depth exploration 24 of the molecular mechanisms of RVA virulence, cell attachment and replication. The specific aims to achieve this goal 25 are summarized below: 26 Aim 1: Evaluate if the D385N and D393H substitutions (individually or in combination with other mutations in the OSU 27 VP4) introduced into OSU RGS (ic-virOSU) will confer an attenuated phenotype to the progeny virus (ic-attOSU). Aim 28 2: Evaluate if G9P[13]-like mutations (S385N and D393N) will confer the G9P[13]-like glycan specificity to the progeny 29 virus (ic-G9OSU). Aim 3: To conduct comparative transcriptome analysis of the wild-type (ic-virOSU) and mutant (OSU- 30 att and OSU-G9) OSU variants. Our PIE and Gn pig models as well as the newly established RGS will allow to study 31 how the identified VP4 mutations alter RVA virulence and the selective interactions with host glycans. 32

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY Millions of people are affected by chronic conditions, and the number will continue to rise due to increased life expectancy. These diseases, including wet age-related macular degeneration (AMD), rely on frequent local injections for disease management. Frequent injections required for maximum therapeutic efficacy are associated with barriers to care including patient discomfort, high treatment costs, and risk of complications. There is a clinical need to reduce injection frequency while maintaining treatment efficacy for these chronic diseases to improve treatment outcomes and quality of life for these patients. The overall objective of this project is to develop a tunable, injectable, biodegradable microcapsule delivery device that has the potential to sustain release of therapeutics of varying molecular weights for at least 12 months. Theoretical modeling will be combined with in vitro and in vivo experimental studies to optimize the microcapsule system and predict therapeutic release. In Aim 1, microcapsule porosity will be modulated using two methods to tune therapeutic release. Release of therapeutics of varying molecular weights, including anti-VEGF (vascular endothelial growth factor) and triamcinolone acetonide (TA), will be evaluated at least 12 months. Therapeutic release rates and bioactivity will be assessed in vitro by evaluating samples using ELISA, Ultra Performance Liquid Chromatography (UPLC), endothelial cell tube formation assays, and inhibition of inflammatory markers in retinal cells. An in silico model for drug release dependent on porosity and drug size will be refined. In Aim 2, microcapsule outer polymer layer thickness will be modulated to tune therapeutic release. Long-term release rates and bioactivity of anti-VEGF and TA will be evaluated in vitro. The in silico model will be refined and validated for drug release dependent on capsule layer thickness. In Aim 3, microcapsule biocompatibility and pharmacokinetic distribution of the therapeutics will be evaluated in vivo. The microcapsule will first be evaluated for 1 month for short-term biocompatibility. Then, fluorescently labeled therapeutics will be loaded into optimized microcapsules and compared to blank capsules and therapeutic only controls for 12 months. Fluorophotometry will be used to quantitatively assess therapeutic concentrations released over time and will be compared to assays conducted on extracted ocular tissues at study termination. Results will be compared to published studies for effective therapeutic concentrations. This study will also provide 12 month in vivo safety data. The in silico model will be refined to include measurements of therapeutic concentrations in the vitreous and retina to predict in vivo distribution coupled to drug release from a capsule. Additionally, a computational tool will be developed for optimizing capsule layer thickness and porosity for specified release duration, pharmacokinetic tissue distribution, and therapeutic size. The goal of this project is to develop a tunable drug delivery device with the potential to reduce injections to one time per year, improving the quality of life for patients with AMD or other chronic diseases that rely on local injections for treatment.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Urothelial carcinoma is a common, aggressive, morbid, and understudied disease. Many patients are not cured with the current standard of care for localized muscle-invasive urothelial carcinoma, which is neoadjuvant combination cisplatin-based chemotherapy followed by radical cystectomy. But overall survival is significantly longer in patients who obtain a pathologic complete response to neoadjuvant chemotherapy, suggesting that intensification of systemic therapy will improve survival. It is critical that we develop reliable predictive biomarkers that can select patients that will or will not have a complete pathologic response to receive, respectively, either chemotherapy or an intensified peri-operative regimen. Preliminary evidence in urothelial carcinoma and other cancers suggests that infiltrating immune cells play a role in treatment response, but this has not been rigorously studied in muscle-invasive urothelial carcinoma. In this study, we will interrogate pre- and post-treatment samples from a completed, pivotal phase II clinical trial, harnessing our expertise in spatial transcriptomics and proteomics to interrogate differential gene and protein expression in tumor, immune, and stromal cells in annotated tissue specimens before and after neoadjuvant chemotherapy with or without a checkpoint inhibitor. Specifically, we will evaluate baseline CD8:FOXP3 ratio and responses to therapy (Aim 1), determine the effect of neoadjuvant therapy on CD8:FOXP3 ratio (Aim 2), and leverage the full capacity of spatial transcriptomic/proteomic assays to develop and evaluate the performance of novel predictive biomarkers of response to neoadjuvant therapy (Aim 3). This study will result in predictive biomarkers while concurrently profiling the immune infiltrate composition and how it is influenced by treatment. The ultimate goal is to design rational combination or sequential peri-operative regimens for biomarker-driven clinical trials to improve patient survival and cure rates. The project will provide the candidate, Katharine Collier, MD, MSc, MSE, MS, with training in rigorous quantitative methods, cutting-edge spatial molecular analyses, and high-dimensional biomarker development. The proposal capitalizes on Dr. Collier’s quantitative background in Chemical Engineering, clinical training in Medical Oncology, and formal training in clinical trial design, while providing an opportunity to gain additional skills and knowledge in multi-omics techniques and data analyses, preclinical studies, leadership, presentations, and grant writing. Dr. Collier is committed long-term to improving outcomes for patients with genitourinary cancers as a translational physician-scientist. Dr. Collier will be supported by an experienced mentorship team (Amir Mortazavi, MD, Zihai Li, MD, PhD, Daniel Stover, MD, Steven Clinton, MD, PhD), skilled collaborators, the rich academic environment of the Ohio State University Comprehensive Cancer Center, and an invested institution committed to providing protected time and resources for career development and research. This award will ensure Dr. Collier’s successful transition to independence as a physician-scientist and translational researcher improving outcomes for patients with genitourinary cancers.

NIH Research Projects · FY 2025 · 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 · 2023-07

SUMMARY/ABSTRACT Heart failure (HF) is a leading cause of death worldwide. Although the pathophysiology of HF is complex and remains incompletely understood, defects in mitochondrial function have been implicated in the progression and outcomes of HF, and emerged as an important target for HF therapy. One well-known contributing factor to adverse cardiac remodeling in the failing hearts is excess mitochondrial-derived reactive oxygen species (mtROS). Accordingly, a variety of antioxidant-based therapies have been developed for HF treatment over the last decades. However, despite promising outcomes in preclinical studies, translation of these therapies to the clinic has not succeeded to date, suggesting that alternative or complementary mitochondrial therapeutic targets are needed. In addition to excess mtROS, profound loss of mitochondrial membrane potential (m) is another key hallmark of HF. In the cardiomyocyte (CM), m disruption affects not only energy production, but also a variety of signaling pathways crucial for cell function and survival, such as redox balance, calcium homeostasis and mitochondrial quality control. We hypothesize that synergistically targeting mitochondria, i.e., concurrently preserving mitochondrial m and scavenging excess mtROS, is a viable therapeutic strategy for HF treatment. However, assessing the therapeutic potential of m preservation is challenging, due to a lack of tools for dynamic and specific control of CM m in live animals. To overcome this technical barrier, we developed an innovative mitochondrial-targeted luminoptogenetic (named mLumOpto) technology by integrating luciferase- luciferin-emitted endogenous bioluminescence with the mitochondrial optogenetics we recently published. Our preliminary data indicate that mLumOpto can induce dynamic CM m control in the absence of external light illumination. The primary goal of this project is to employ this advanced mLumOpto technology to manipulate CM m in vivo to evaluate the efficacy of CM m preservation alone, or in synergism with a mtROS scavenger (i.e., mitochondrial-specific antioxidant), in HF treatment in preclinical mouse models. The translational potential of the proposed synergistic mitochondrial-targeted HF therapy will also be assessed in human-like large animals (i.e., pigs). Three Specific Aims are proposed to accomplish our objectives. Aim 1 will determine the role of sustained CM m depolarization in pathological cardiac remodeling and HF development in mice, and dissect the underlying molecular mechanisms. Aim 2 will evaluate the efficacy of synergistic m preservation and mtROS scavenging in improving adverse cardiac remodeling and contractile dysfunction in two well-established mouse HF models (i.e., pressure overload and myocardial ischemia-reperfusion). Aim 3 will assess the translational potential of mLumOpto-mediated HF therapy in pigs. Successful completion of this project will lead to not only an innovative technology capable of specifically and dynamically manipulating CM mitochondria in vivo, but also a novel translational mitochondrial-targeted therapy for HF treatment.

NIH Research Projects · FY 2025 · 2023-07

Heart disease is the leading cause of mortality worldwide and disruption of transcription in cardiomyocytes (CMs) contributes to the progression of heart disease. Transcriptional regulation of some individual genes by transcription factors (TFs) is described. However, much less is known about how functionally related genes, or gene expression programs, are collectively regulated in a coordinated manner in healthy and diseased CMs. The Mediator Complex serves as a bridge to link transcriptional machinery and TFs to control transcription, but the molecular mechanisms of transcriptional regulation by Mediator are not well-understood. Mediator is a large multiprotein complex organized as submodules, with the CDK8 kinase submodule regulating RNA Polymerase II transcriptional activity. MED12 is an essential Mediator component, and within the kinase submodule, is required for CDK8 kinase activity. Although traditionally considered a transcriptional repressor via the CDK8 kinase submodule, our published data demonstrates that increased MED12 is also able to directly activate transcription. Our preliminary studies indicate that MED12 levels are increased in failing hearts from humans and mice, sup- porting the clinical relevance of this novel MED12 function in mis-regulation of transcription in heart disease. Our published studies in mice with CM-specific Med12 deletion demonstrate that MED12 regulates a calcium-handling gene expression program, through direct interaction with the TF MEF2. We identified additional TFs that interact with MED12, and these findings suggest that MED12 has multiple transcriptional functions depending on its interactions, but its functions have not been delineated. To determine the contribution of MED12 to transcriptional mis-regulation in heart disease, it is crucial to define its molecular interactions and functions in CMs and identify its cell-specific roles. Subpopulations of kinase submodules have been reported in other cells, but their functions have not been defined. Additionally, functions of individual kinase submodule proteins have not been investigated for their roles in alternative kinase subcomplexes or for functions independent of the kinase submodule. Our published CM-specific Med12 knockout model (cKO) and newly generated CM Med12 transgenic model (cTg) both develop heart failure, emphasizing the critical contribution of MED12 to transcriptional homeostasis in CMs. We will use our Med12 genetic models to delineate the functions of MED12 in CMs. We will identify MED12 kinase subpopulations, interactions with TFs, and genomic interactions in normal CMs and those with MED12 overexpression. We will also assess the ability of a human MED12 missense mutations to bind TFs and kinase submodule proteins using hi-PSCs. The long-term goal of my lab is to uncover mechanisms that control coordinated transcriptional programs in the heart. In this proposal we aim to 1) determine the mechanisms of MED12-regulated transcription in CMs, and 2) identify the transcriptional mechanisms of MED12- driven heart failure. Consequently, a better understanding of the molecular mechanisms governing cardiac gene expression will provide new insight into therapeutic strategies to restore cardiac function during disease.

NIH Research Projects · FY 2025 · 2023-07

Project Summary / Abstract The graduate program in vision science at The Ohio State University has a long track record of training optometrists for careers as clinician scientists. The program has yielded many NIH funded investigators, clinical trialists, and significant contributors to the field of vision science. More recent partnerships with ophthalmology and neuroscience have only further strengthened the program which boasts a breadth of research training opportunities from basic science, animal models, patient-based research, and multi-center clinical trials. This proposal seeks funding to provide support for post-doctoral trainees who have completed an optometric doctorate and are now seeking a PhD. The proposal seeks to support one trainee in the initial year and add an additional trainee in each of years 2 & 3 to reach a total number of three trainees for the third and subsequent years. The faculty appointed to this award have strong records of NEI funding and provide a rich variety of research topics to include corneal biomechanics and wound healing, retinal regeneration, refractive error, pediatric and binocular vision, population vision health, and low vision rehabilitation. Trainees will gain education in the fundamentals of vision science and the ethical conduct of research, complete mock grant proposals, write a review paper, run research projects under the mentorship of their advisors, participate in public presentation of their research at least annually, and be mentored in the publication of their scientific work. Engaging optometrists in rigorous scientific training is critical to maintain the workforce needed to address the many vision disorders impacting both the aging population and our youth. Ohio State University is one of the leading institutions in yielding clinician scientists from the pool of US trained optometrists. The support of this award will further enhance the ability of the vision science graduate program to produce excellent researchers by providing stipends to maximize the dedicated research time of the trainees and focus their efforts solely on training.

NIH Research Projects · FY 2025 · 2023-07

Abstract: ARDS accounts for one in ten intensive care unit (ICU) admissions. Mortality in ARDS is estimated at 30 – 50%. Decades of clinical trials have failed to identify targeted pharmacotherapeutics that significantly impact disease course. New insights are needed to broaden our understanding of the underlying mechanisms that lead to ARDS to promote the development of novel therapeutics. Transfer RNA (tRNA) constitutes 10 – 20% of the total cellular RNA content. Although classically thought of passive translational machinery, more recent studies show that tRNA can be dynamically regulated in response to the environment. The post-translation modification of tRNA by methylation is one mechanism by which tRNAs respond to cellular stress. Methylation of tRNAs can alter translation, change the distinction between ‘self’ and ‘non-self’, and protect tRNAs from fragmentation. Despite this, the role of tRNAs in infection and critical illness is unknown. We identified a methyltransferase enzyme, termed TRMT1 that is upregulated in the human lung in response to infection. We also find that the methylation of tRNA by TRMT1 is altered in response to inflammatory stimuli. Our preliminary data suggest that the TRMT1 is vital for macrophage viability and host defense in infection, and for maintaining cytokine responses. This application focuses on the role of the TRMT1 in acute lung injury and the investigation of tRNA biology, unexplored territory in ARDS and critical care illnesses. During the execution of these studies, we will examine how myeloid TRMT1 contributes to immunopathology in experimental lung injury. We will determine how TRMT1 alters inflammatory signaling and regulates cell death in vitro and in vivo. To examine the mechanistic role of TRMT1, we will measure the methylation of tRNA in response to inflammatory stimuli and the production of tRNA fragments. To further explore the regulation of TRMT1, we will examine how subcellular trafficking of TRMT1 differentially regulates cytoplasmic and mitochondrial tRNA methylation and how this alters the inflammatory response. Finally, because cellular concentrations of TRMT1 protein may be crucial in regulating macrophage behavior and viability we will examine the E3 ligase mediated ubiquitination and degradation of TRMT1. These studies will be the first to elucidate the role of TRMT1 in acute lung injury. In addition, these studies will provide new insight into the role tRNA methylation in lung injury. The execution of these studies will provide significant mechanistic and biological advances in the field of acute lung injury and ARDS and lead to novel interventional targets for therapeutics.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY The global rate of HIV infection and the number of AIDS related deaths have dramatically declined due to the expanding access to combinatorial anti-retroviral therapy (cART). However, the HIV epidemic remains and there is still no cure for HIV infection. Because cART does not eradicate HIV, while replication-competent viruses become integrated and silenced proviruses, which can be sporadically reactivated to replenish viral reservoirs. As a key anatomical “sanctuary site” for HIV infection and persistence/latency, reservoirs in brain remains a main hurdle for HIV cure. HIV-infected monocytes contribute to viral spread from the periphery blood to the brain as they can migrate across the blood brain barrier (BBB) and differentiate into microglia in the central nervous system (CNS). Microglia are a major and stable viral reservoir for persistent/latent HIV infection in the CNS, even in the presence of cART. Furthermore, HIV-infected microglia release neurotoxic factors that promote neuroinflammation and contribute to HIV-associated neurocognitive disorders (HAND). Therefore, characterization of molecular features regulating HIV persistent/latent infection in microglia is essential for developing effective intervention strategies to control and inhibit HIV in the CNS. Epigenetic regulation critically determines the faith of HIV proviruses and contributes significantly to HIV latency. The focus of this project is to investigate the role of host epigenetic regulation in promoting persistent/latent HIV infection in microglia and target it for preventing HIV lytic reactivation. We have identified a set of Jumonji domain-containing histone lysine demethylases (KDMs) as HIV latency-promoting genes (LPGs) that preferentially target histone H3K4/K36 methylation (H3K4/K36me3), which indicates the potential role of histone demethylation in regulation of HIV latency. In Aim 1, we will investigate the contribution of these KDMs to promoting HIV latency in human microglia and iPSC-derived microglia containing organoids (MCOs). In Aim 2, encouraged by the KDM studies, we would like to continue the use of our functional genomic and proteomic expertise to identify other ovel host factors governing HIV latency in microglia by multidisciplinary approaches, including single-cell CRISPR screen. This is an important issue to address so that we can novel host targets to develop more potent regimen for reinforcing HIV latency and preventing its resurrection at a cART-free setting in microglia. Furthermore, we also screened a library of FDA-approved drugs and identify HIV latency- promoting agents (LPAs), including levosimendan (LSM). In Aim 3, we propose the translational studies to investigate the repurposing of FDA-approved drugs already used in clinic for treating and blocking HIV persistent/latent infection in microglia, which likely affect epigenetic processes.

NIH Research Projects · FY 2025 · 2023-07

PROJECT ABSTRACT Colorectal cancer (CRC) is the fourth most common cancer, and the second leading cause of cancer death in the United States, with an estimated incidence of 151,030 new cases in 2022. According to the American Cancer Society, the lifetime risk of developing colorectal cancer is 1 in 23 for men and 1 in 25 for women. Tumor budding is a prognostic factor in colorectal cancer with potential to risk stratify patients and possibly guide treatment decisions. It is defined as the presence of a single tumor cell or a cell cluster consisting of fewer than five tumor cells at the invasive tumor front. Unfortunately, tumor budding is not routinely disclosed in pathology reports due to lack of reproducible methods in identifying tumor buds from H&E slides. The prevalence, mortality, and risk of colorectal cancer as well as the potential of tumor budding as a prognostic factor necessitate an accurate, easy- to-use, reproducible system to identify tumor budding. We aim to develop a computer-aided image analysis system to standardize the quantitative criteria used to define tumor budding from H&E slides. In addition to identifying tumor buds, the system will correlate tumor buds with several outcomes (microsatellite instability status, overall survival, progression free survival, and disease free survival). As part of the proposed computer- aided image analysis system, we will first develop a sophisticated method for color deconvolution to compensate for color variations. This will be followed by deformable image registration and deep learning modules to differentiate tumor from non-tumor regions. The study will show that machines can be trained using deep learning to identify different anatomical regions within H&E slides of colorectal patients. From thereon, we will rely on scale-space theory and alpha-shapes to identify tumor buds and hotspots. We will use mathematical morphology and differential geometry to extract visually meaningful imaging features from tumor buds and hotspots. We will explore the potential of these imaging features along with features produced by our unsupervised multiple instance learning in predicting outcomes. The proposed research will help identify the association of tumor budding to colorectal cancer outcomes. The model will be subjected to rigorous statistical analysis for accuracy and reproducibility. The project will result in innovative software tools that facilitate the selection for personalized cancer therapies for colorectal patients.

NIH Research Projects · FY 2026 · 2023-07

PROJECT SUMMARY The global rate of HIV infection and the number of AIDS related deaths have dramatically declined due to the expanding access to combinatorial anti-retroviral therapy (cART). However, the HIV epidemic remains and there is still no cure for HIV infection. Because cART does not eradicate HIV, while replication-competent viruses become integrated and silenced proviruses, which can be sporadically reactivated to replenish viral reservoirs. As a key anatomical “sanctuary site” for HIV infection and persistence/latency, reservoirs in brain remains a main hurdle for HIV cure. HIV-infected monocytes contribute to viral spread from the periphery blood to the brain as they can migrate across the blood brain barrier (BBB) and differentiate into microglia in the central nervous system (CNS). Microglia are a major and stable viral reservoir for persistent/latent HIV infection in the CNS, even in the presence of cART. Furthermore, HIV-infected microglia release neurotoxic factors that promote neuroinflammation and contribute to HIV-associated neurocognitive disorders (HAND). Therefore, characterization of molecular features regulating HIV persistent/latent infection in microglia is essential for developing effective intervention strategies to control and inhibit HIV in the CNS. Epigenetic regulation critically determines the faith of HIV proviruses and contributes significantly to HIV latency. The focus of this project is to investigate the role of host epigenetic regulation in promoting persistent/latent HIV infection in microglia and target it for preventing HIV lytic reactivation. We have identified a set of Jumonji domain-containing histone lysine demethylases (KDMs) as HIV latency-promoting genes (LPGs) that preferentially target histone H3K4/K36 methylation (H3K4/K36me3), which indicates the potential role of histone demethylation in regulation of HIV latency. In Aim 1, we will investigate the contribution of these KDMs to promoting HIV latency in human microglia and iPSC-derived microglia containing organoids (MCOs). In Aim 2, encouraged by the KDM studies, we would like to continue the use of our functional genomic and proteomic expertise to identify other ovel host factors governing HIV latency in microglia by multidisciplinary approaches, including single-cell CRISPR screen. This is an important issue to address so that we can novel host targets to develop more potent regimen for reinforcing HIV latency and preventing its resurrection at a cART-free setting in microglia. Furthermore, we also screened a library of FDA-approved drugs and identify HIV latency- promoting agents (LPAs), including levosimendan (LSM). In Aim 3, we propose the translational studies to investigate the repurposing of FDA-approved drugs already used in clinic for treating and blocking HIV persistent/latent infection in microglia, which likely affect epigenetic processes.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY As the global leading cause of death, heart failure is a major challenge for researchers in their quest to discover therapeutics that can save countless lives. After cardiac injury, the heart begins to remodel itself in a way that is initially adaptive, but this innate coping mechanism may over time expedite heart failure onset. Elucidating the mechanisms which underly the progression from adaptive cardiac hypertrophic remodeling to heart failure will dramatically impact the discovery of novel therapeutics for this deadly disease. While regulation of gene expression through transcription of messenger RNA (mRNA) has been extensively studied, only recently an appreciation for the importance of chemical modifications that can occur on mRNA has emerged. This proposal focuses on the methylation of the N6-Adenosine of mRNA (m6A), which is the most abundant internal mRNA modification in eukaryotes. Previous research from our lab has shown that modulation of m6A content in the heart is sufficient to drive cardiac remodeling and to affect the ability of the heart to respond to stress. Despite this, the exact mechanisms through which this occurs is not well understood. The fate of m6A-modified mRNAs is regulated by members of the YTH Domain Family (YTHDF). We found that YTHDF3 is specifically important in cardiomyocytes, where it localizes to the nucleus and binds to Myocyte Enhancer Factor 2D (MEF2D), which is an important transcription factor regulating hypertrophic cardiac growth. Further, we have found that knockout of YTHDF3 mitigates pathological remodeling following pressure overload injury. Given these preliminary data, we hypothesize that YTHDF3 regulates cardiomyocyte size and stress-induced remodeling by modulating the processing of m6A-modified mRNAs transcribed by MEF2D. To test this hypothesis, we already generated and validated a new mouse line in which YTHDF3 can be selectively deleted in cardiomyocytes (YTHDF3-cKO). In Aim 1, we will investigate the role of YTHDF3 at baseline and in the stressed murine heart using longitudinal echocardiography analysis, and assessing histological and molecular signs of pathology at the terminal time point. In Aim 2, we will determine the mechanism through which YTHDF3 regulates the fate of specific subsets of MEF2D-transcribed m6A-mRNAs in cardiomyocytes. First, we will further characterize the binding between YTHDF3 and MEF2D by defining the respective domains involved. Then, we will dissect the binding of YTHDF3 to MEF2D mRNA targets and determine consequent stability, export, and translation of these transcripts. Finally, in Aim 3, we will undertake an unbiased approach to more globally investigate the role of YTHDF3 in regulating mRNA biology in healthy and stressed adult cardiomyocytes by cross-linking immunoprecipitations of YTHDF3-bound mRNAs followed by sequencing (CLIP-seq). Our approach is innovative and significant, as it will be the first project to define the role of YTHDF3 in the heart, which may lead to a new field of therapeutics based on the biology of mRNA methylation.

NIH Research Projects · FY 2024 · 2023-07

Project Summary Our proposed T32 training program in health services research (HSR) at The Ohio State University (OSU), “ACCELERATE: Accelerating HSR at OSU,” will prepare the next generation of researchers and clinician researchers for careers in health services and health systems research. Each year we will recruit four postdoctoral trainees from research and clinically oriented backgrounds to participate in our collaborative, multidisciplinary training program. Our goal is to have a diverse pool of recruits who bring a wide range of backgrounds and experiences to our program. Trainees will enroll in a two-year program designed to provide individualized mentoring and training to prepare participants for independent careers as health services researchers. Training elements will include a strong experiential component of health services and health systems research content and methods alongside didactic instruction in relevant topics. Improving health equity and increasing diversity and inclusion will be central themes throughout the program and related curriculum. Together, trainee-mentorship teams will create individualized development plans focused on the trainee’s areas of interest and career goals. Trainees will be expected to disseminate their research through publications and presentations at national conferences, participate in the annual NRSA trainees research conference, and prepare a competitive research proposal by the end of their program. The ACCELERATE T32 Program will be led by a team of three experienced principal investigators who are nationally recognized in health services and health systems research. Three additional co-investigators will provide supplemental methodological and HSR content expertise. An Advisory Committee comprised of ten senior HSR leaders from across OSU will provide strategic direction for program development and evaluation. Unique aspects of ACCELERATE include being anchored in one of the most diverse colleges of medicine in the country, capitalizing on the unparalleled depth and breadth of academic programs at OSU, focusing on developing mentorship skills of junior faculty in partnership with experienced senior mentors to build the pipeline of HSR mentors, and recruiting and training both non-clinician and clinician researchers to study and tackle the myriad important challenges in HSR. Our goal is to train health services researchers to address the most pressing issues in healthcare delivery, using data and innovative processes to help people increase their well- being and access affordable, safe, and effective care throughout their lives.

NIH Research Projects · FY 2026 · 2023-07

Project Summary/Abstract While research on substance using youth experiencing homelessness (YEH) is increasing, there is a dearth of information regarding effective prevention interventions for these youth. This is of significant concern because studies indicate that 66% to 89% of YEH have a mental health disorder (Cauce et al., 2004) and 68% report at least one suicide attempt (Gewitz et al., 2020). In fact, suicide is the leading cause of death among YEH (Rotheram-Borus & Milburn, 2004; Yoder et al., 2010). Among those who have attempted suicide, an average of 6.2 attempts is reported, and lifetime suicidal ideation rates range from 14% to 66.5% (Yoder et al., 2010) with no clear evidence of effective interventions for YEH. This study uses general cognitive theory (Beck, 1967), complemented with concepts from two suicide specific theoretical models, to guide our intervention and conceptual change model. Our goal is to test strategies to extend delivery and uptake of a cognitive therapy intervention for YEH. Prior studies focus on service-connected youth, but research suggests that most street living YEH do not access services meant to assist them (Kelly & Caputo, 2007). We overcome prior sampling limitations through engaging service-disconnected youth in addition to service-connected youth through a local drop-in center. As such, three hundred substance using YEH with recent suicidal ideation or a recent suicide attempt will be randomly assigned to Cognitive Therapy for Suicide Prevention (CTSP) + Services as Usual (SAU) (N=150) or to SAU alone (N=150). CTSP has previously shown efficacy for YEH through a RCT pilot feasibility study (Slesnick et al., 2020; Wu et al., 2020). SAU includes outreach, advocacy and service linkage which are typical services offered by agencies serving those experiencing homelessness (Ober et al., 2012). Follow-up assessments will be conducted at 3, 6, 9 and 12-months post-baseline. It is hypothesized that youth receiving CTSP+SAU will show greater reductions in suicidal ideation (primary outcome), substance use and depressive symptoms (secondary outcomes) over time compared to SAU alone, as well as improved risk and protective factors. Theoretically-derived mediators will be tested to shed light on mechanisms associated with change, and the moderating effects of sex, race, sexual orientation and baseline service connection will be examined. In order to ease future dissemination of the intervention to agencies serving YEH, we will rigorously assess acceptability, feasibility, fidelity and cost associated with the delivery of our intervention approach using a mixed-methods approach. Ultimately, the goal of this research is to provide support for the use of a suicide prevention intervention for substance using YEH that reduces premature mortality, hospitalization, and loss of human capital and which can be easily adopted by agencies serving YEH.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY/ABSTRACT: This K01 Mentored Research Scientist Development Award will provide the candidate, Sehun Oh, PhD., with protected time and resources to facilitate his transition to research independence in substance use and prevention research. Although we have been observing disconcerting rates of substance-related problems, including overdose (OD) deaths, in communities with declining economic opportunities (especially in the Appalachian and Rust Belt areas), we do not have firm theoretical and empirical evidence to explain how local labor market contexts may affect substance use (SU) and development of substance use disorder (SUD) and OD as well as what effective policy interventions might reverse the trends. Building on his interdisciplinary background in economics, substance use epidemiology, and social policy, the candidate will engage in structured training and research activities to acquire substantive and methodological expertise to elucidate the role of local labor market contexts in SU/SUD/OD. Specifically, the proposed career development plan is designed to achieve three training goals: (1) deepening the candidate’s understanding of local economic contexts (especially the types and qualities of available jobs) as a key etiology of substance use problems and its socioeconomic, physical, and behavioral health mechanisms; (2) developing expertise in agent-based modeling and supporting methods (causal inference and spatial analysis) for local SU/SUD/OD projections; and (3) gain hands-on experiences in research translation and dissemination to inform prevention policy development. A series of training activities have been carefully designed, including coursework in epidemiology, public policy, developmental economics and social work; individualized instruction; mentored training; and research collaborations over five years. This training also will be achieved by conducting a minimum of three research studies that address the following aims: (1) elucidating the role of the local labor market (re)structuring in SU/SUD among individuals in young/middle adulthood, (2) developing agent-based models to make SU/SUD/OD projections in Ohio until 2030 and to conduct policy experiments to assess the preventive effects of employment and substance supply-related interventions. To address these aims, the candidate will integrate and analyze various administrative and survey data (related to local labor market contexts, SU/SUD/OD, and other environmental characteristics); test the effects of industry-specific job availability (and its changes) and socioeconomic/physical/behavioral health mediators on SU/SUD/OD; develop agent-based modeling to make robust projections in Ohio based on local labor market outlook; and conduct simulated experiments to assess the preventive impacts of employment- and substance supply-related policies using agent-based models. By the end of the award period, the candidate will be positioned as one of the few investigators in the United States with the requisite knowledge and skills to use systems science and simulation modeling to inform substance use prevention policies, especially for communities experiencing economic restructuring and/or contractions.

NIH Research Projects · FY 2026 · 2023-07

Cannabis use disorder (CUD) is most prevalent in young adulthood (ages 18-25), with 47% of CUDs developing during this period. Alarmingly, the proportion of young adults who use cannabis on a daily basis nearly doubled from 2013-2021, which is linked to a 17-fold increase in risk for CUD. Combined with the paucity of interventions for CUD, this highlights the urgent need for CUD research among young adults. Examining event-level risk processes clarifies the temporal order of risk factors and cannabis use (CU) as they unfold in near-real time. However, ecological momentary assessment (EMA) studies, including our own, provide mixed evidence for two sets of risk factors for CU: risk factors related to increasing positive affect (positive reinforcement; e.g., enhancement motives) and decreasing negative affect (negative reinforcement; e.g., coping motives). Providing a potential explanation for mixed evidence, our preliminary results and the multistage model of substance use suggest these two sets of risk factors may be relevant at different stages of CUD, with positive reinforcement most relevant when CUD symptoms are absent to mild and negative reinforcement becoming prominent when CUD symptoms become more severe. We propose to test the multistage model using a measurement burst design, with 5 semi-annual bursts of 14-days of EMA (2 surveys per day), allowing us to examine changes in event-level associations between risk factors and CU as a function of longer-term (i.e., semi-annual) changes in CUD symptoms. While risk processes discussed previously unfold at the event-level, these processes contribute to broader patterns of CU frequency. These patterns of change in CU result in trajectories of CU frequency, which vary across individuals. For example, some individuals may start with a low CU frequency that increases over time, while others may maintain a high CU frequency over time. These patterns of CU frequency are theorized to be predictive of the development of CUD. Therefore, we propose to use semi-annual longitudinal data to identify multiple trajectories of CU frequency and examine differences across trajectories in the likelihood of having a CUD diagnosis at the final wave of data collection. This will help to inform preventive efforts by identifying individuals likely to develop CUD. The purpose of this R01 is to advance our understanding of CUD, with a focus on how event-level associations between risk factors and CU change as CUD develops. We will use a measurement burst design, with 600 young adults (age 18-25) who use cannabis at least four times in the past month at the screening survey. The proposed study will accomplish three specific aims: 1) identify changes in the effects of positive reinforcement event-level risk factors on CU as CUD develops, 2) test for changes in the effects of negative reinforcement event-level risk factors on CU as CUD develops, and 3) examine associations between trajectories of CUD symptoms over a 2 year period and the likelihood of having a CUD at the final wave of data collection. The proposed study will inform efforts to increase efficacy of existing CUD interventions.