Objectives/Goals: To evaluate the impact of social determinants of health (SDOH), specifically socioeconomic status and medical insurance coverage, on access to mental health services for adults aged 60 and older served by Federally Qualified Health Centers (FQHCs) in Puerto Rico. Methods/Study Population: A secondary retrospective analysis of electronic health records from FQHCs in Puerto Rico will be conducted to examine the relationship between SDOH needs and mental health service utilization among adults aged 60 years and older receiving primary care. SDOH data will be collected using the PRAPARE® tool, assessing factors such as socioeconomic status, insurance coverage, and emotional support, with a focus on identifying unmet needs. Multivariable and logistic regression models, using Stata v.17, will be employed to evaluate correlations between these SDOH factors and mental health service utilization, adjusting for age, gender, and mental health conditions. This analysis aims to quantify the impact of SDOH on access to mental health services and elucidate key barriers to care for older adults in Puerto Rico. Results/Anticipated Results: We anticipate that lower socioeconomic status, lack of supplemental insurance, and inadequate family support will be strongly associated with the underutilization of mental health services among older adults in Puerto Rico. These disparities are expected to be more pronounced in individuals with limited income, weaker family networks, leading to significant gaps in access to necessary mental health care. Barriers such as healthcare insecurity, financial hardship, and fragmented support systems will likely emerge as major obstacles. By including patients from both rural and urban regions, the study will capture the distinct challenges each population faces, enhancing the contextual relevance of the findings to broader populations, ultimately informing policy and developing intervention strategies. Discussion/Significance of Impact: The findings will provide crucial insights for developing targeted interventions to enhance mental health care access for older adults in Puerto Rico. These results will inform policy development and public health strategies, addressing disparities and promoting equitable care in underserved populations.

Objectives/Goals: The overarching objective of this study is to inform clinicians, patients, and other stakeholders of the level of evidence and the real-world risk-to-benefit profiles associated with older antidepressant drugs that are frequently used off-label. Methods/Study Population: A PubMed literature review was performed to identify clinical trials conducted in the USA between 2013 and 2023 for trazodone and 2000 and 2023 for escitalopram and citalopram. These studies were examined for robustness, due to sample size, study design, and generalizability. Findings were compared with information provided on UpToDate® LexiDrug™, a primary database used by clinicians to inform prescribing practice. To explore risks associated with off-label use, the FDA adverse event reporting system was probed to identify adverse events reported for each drug; results were systematically categorized by reason for use. To compare the volume of on-label to off-label prescriptions, data will be extracted from electronic health records from University of Southern California-affiliated hospitals. Results/Anticipated Results: Studies conducted on off-label prescriptions of these drugs show primarily small sample sizes, pointing to a limitation in generalizability. For citalopram (N = 77) and trazodone (N = 42), over half of their off-label studies had samples of 50 participants or less. These two drugs also showed low evidence rating for off-label prescription on LexiDrug due to limited power studies. Multiple health agencies recommend against off-label prescriptions for trazodone due to insufficient evidence. There is limited data in the US regarding the volume of off-label prescriptions; however, trazodone’s FAERS analysis indicated a large proportion of adverse event reports (1099/7239) come from cases where trazodone was used for insomnia, an off-label indication, compared to depression, the on-label indication (464/7239). Discussion/Significance of Impact: With 1 in 6 Americans taking antidepressants and 40%–80% of these psychiatric prescriptions being employed off-label, there is a serious and present risk for patients regarding the safety and efficacy of these medications. Awareness must be brought to clinicians to protect patients and encourage evidence-based practice.

Objectives/Goals: Transgender women who have sex with men (TGWSM) have higher HIV risk. The rectal mucosal (RM) immune environment of TGWSM who choose feminizing hormone therapy (FHT) has been shown to be distinct from the RM of cisgender men who have sex with men (MSM). We studied the impact of FHT on the adaptive immune cellular composition of the RM. Methods/Study Population: We sampled cross-sectional and longitudinal cohorts of TGWSM and cisgender MSM from The Silom Clinic in Bangkok, Thailand from December 2020 to December 2023. We included participants aged >18 years, all cisgender MSM and TGWSM with FHT levels in the therapeutic range for cisgender women. We performed RM biopsies and analyzed the adaptive immune cell characteristics via flow cytometry. We will perform binary linear regression to assess the association between systemic FHT levels and the percentage of CD4+ T cells expressing key biomarkers. Primary outcomes include the percentage of CD4+ T cells that express CCR5, with a secondary outcome of the percentage of CD4+ T cells that express Ki67. Results/Anticipated Results: The cross-sectional cohort included 100 TGWSM on FHT and 50 cisgender MSM. The longitudinal cohort included 25 TGWSM who were initiating FHT. Similar primary and secondary outcomes are to be elucidated in both cohorts. We anticipate the RM environment of TGWSM using FHT in both cohorts compared to the RM environment of cisgender MSM in the cross-sectional cohort will be associated with greater percentages of activation/co-receptor expression of CD4+ T cells that express biomarkers of interest. In the longitudinal cohort, we similarly anticipate increased percentages and activation/co-receptor expression of CD4+ T cells expressing biomarkers of interest in TGWSM after in comparison to before initiating FHT. Discussion/Significance of Impact: This is the largest study of its kind to compare HIV target cells in RM of TGSWM, which challenges prevailing perspectives suggesting to group cisgender MSM with TGWSM. Anticipated results will inform HIV prevention strategies and future vaccine studies in this high-risk population.

Objectives/Goals: Although eHealth tools like websites, apps, and wearables are widely available, underserved groups often do not benefit equally. This gap is due to usability challenges and overlooked structural, physical, and psychosocial barriers. Additionally, high costs and licensing issues make these tools hard to sustain and share. Methods/Study Population: This case study presents lessons learned over eight years of designing and disseminating a user-centered educational website on human papillomavirus (HPV) and the HPV vaccine for Hmong parents and teens (Hmong Promoting Vaccines, www.hmonghpv.com [http://www.hmonghpv.com]), a community-based participatory research project. Results/Anticipated Results: Our community-driven approach revealed four key principles for creating and sustaining culturally tailored eHealth tools for underserved groups: * Engage stakeholders like community members, legal teams, and developers early and keep them involved. * Discuss dissemination and sustainability goals from the start. * Explore commercialization options, balancing sustainability with protection for underserved groups. * Identify and use academic resources to discuss commercialization, ownership, copyright, and intellectual property of such eHealth interventions. Discussion/Significance of Impact: The Hmong Promoting Vaccines case study highlights the need for an inclusive approach to designing sustainable eHealth tools for underserved communities. Early stakeholder engagement, careful planning for dissemination, and balancing commercialization with protection can reduce health disparities and create fairer digital solutions.

Objectives/Goals: Ischemic stroke treatments assist in restoring blood flow, but do not guarantee good outcomes. Since extracellular vesicles (EVs) able to cross the blood brain barrier, total (nonspecific) and astrocyte enriched EVs (TEVs, AEVs, respectively) from plasma may emerge as plasma biomarkers for prognostication and targeted therapeutics. Methods/Study Population: “Blood and Clot Thrombectomy Registry and Collaboration” (BACTRAC; NCT03153683) is a human stroke biobank at the University of Kentucky that collects samples at the time of mechanical thrombectomy during emergent large vessel occlusions (ELVO; ischemic stroke). EVs were isolated, via size exclusion chromatography, from unbanked plasma and concentrated resulting in TEVs. AEVs were immunoprecipitated with anti-EAAT1 (GLAST), an astrocyte-specific transmembrane glycoprotein. Isolated protein was sent to Olink and ran on their metabolic panel. Demographics and medical histories of the subjects were exported from REDcap and investigators were blinded during EV analysis. Results/Anticipated Results: ELVO subjects (8 females/ 5 males) were an average age of 71.1 ± 11.7 years. Lower TEV enolase 2, a neuronal glycolysis enzyme, associated with increased stroke severity (NIHSS; rs =  -0.7819, p = 0.0476). Higher systemically TEV quinoid dihydropteridine reductase (QDPR), essential co-factor enzyme, was associated with more severe strokes (NIHSS; rs =  0.8486, p = 0.0123) and lower cognition (MoCA; r2 =  0.7515, p = 0.0254). Interestingly, higher intracranial AEVs QDPR was associated with lower infarct volumes (rs =  -0.7333, p = 0.0202), less severe strokes (NIHSS; rs =  -0.6095, p = 0.0388), and better cognition (MoCA; r2 =  0.6095, p = 0.0388). Increased AEV nicotinamide adenine dinucleotide kinase another essential co-factor enzyme, intracranially also correlated to higher cognition (MoCA; rs =  0.8356, p = 0.0298). Discussion/Significance of Impact: Plasma TEV and AEV metabolic proteins correlate with the progression of stroke outcomes and should be investigated as target therapies during MT to improve outcomes.

Objectives/Goals: The goal of this proposal is to better understand how informing African Americans of their genetic risk affects their behavior as part of a cardiovascular disease (CVD) risk reduction intervention. Aim 1: To determine the effect of genetic risk knowledge on CVD health behavior. Aim 2: To determine the effect of genetic risk knowledge on secondary variables. Methods/Study Population: Method: Fifty participants from the Baton Rouge metropolitan area will be recruited. Participants must be African American adults over the age of 18. Potential participants will be recruited using community-based efforts that have been successful in recruiting this population specifically. Participants will be randomized into one of two groups. Genetically Unblinded Group (GU) will be “genetically unblinded” after baseline orientation. Genetically Blinded Group (GB) will be “genetically blinded” until the end of the study. This study design ensures that we can measure the impact of knowledge of genetic risk on participant behavior. Results/Anticipated Results: Baseline participants’ characteristics (body mass index, blood glucose, and cholesterol) will be summarized by intervention group, with counts and percentages for categorical variables and means and 95% confidence intervals for continuous variables. Primary Outcomes: Attendance in intervention sessions will be counted across groups. Effect on genetic risk knowledge will be determined via comparing the difference between the increased healthy lifestyle behaviors at endpoint between Genetically Unblinded (Cases) and Genetically Blinded Groups (Control). Secondary and Tertiary Outcomes: Mean change in secondary outcomes in the GU group will be compared against the mean change in the GB group. Participant’s survey responses and changes in physical measurement from baseline to endpoint will be observed. Discussion/Significance of Impact: This study empowers African Americans in Baton Rouge by providing genetic risk knowledge for cardiovascular disease. By addressing social determinants of health, it promotes behavior change, improves health outcomes, and fosters trust, potentially reducing health disparities and advancing health equity.

Objectives/Goals: Clinical research professional (CRP) managers often struggle to onboard effectively in the rapid timelines necessary. We developed a competency-based, standardized, onboarding program using a 3-factor adult learning approach and designed to be easily adaptable, broadly shared, and readily implemented across a variety of research environments. Methods/Study Population: The Duke competency-based onboarding program for CRPs was developed through an iterative process with input from research community members. Initially, 97 courses were mapped to clinical research competencies using the Joint Taskforce for Clinical Trial Competency framework to identify training gaps and establish a structured learning framework. The onboarding program includes three key components: role specific Express Start modules (self-paced e-learning), Onboarding Learning Plans (a customizable timeline for competency-mapped trainings), and Engagement Activity Packets (guides for manager-driven applied learning in specific competency areas). An additional cohort-based mentorship program (RPN+) includes 4 months of mentored group learning and seminars designed for new professionals. Results/Anticipated Results: Since launch in 2021, 521 new employees have registered an Onboarding Learning Plan representing more than 55% of new CRPs. Additionally, nearly 85% of new CRPs have completed the Express Start role-specific online modules. 54% of new CRPs have enrolled in RPN+ (launched in 2022) and have included members from 22 of 24 clinical research units. Users have reflected the range of clinical research staff roles at Duke, with the majority being clinical research coordinators. A significant predominance of participants found the programs beneficial and would recommend to others. Since launch, program materials have been shared with 64 external groups and institutions via 91 requests. Discussion/Significance of Impact: Duke’s CRP onboarding program addresses organizational, technical, and social aspects through Express Start, Onboarding Learning Plans, and the RPN+ mentoring program. Aligned with the JTFCTC framework, and designed for broad implementation, it successfully promotes competency-based growth and optimizes time for both managers and employees.

Objectives/Goals: This study investigates the contribution of the stool secretome (the soluble factors secreted by microbes into extracellular space) to in vitro α-synuclein (αSyn) oligomerization using stool cultures from patients with multiple system atrophy (MSA), a rare neurodegenerative disease hallmarked by pathologic αSyn aggregates. Methods/Study Population: Stool samples from MSA patients (n = 20), household controls (n = 20), and healthy controls (n = 20) will be cultured using an adapted dilution-to-extinction approach. The goal is to reduce microbial complexity progressively to produce random secretome combinations that may affect αSyn oligomerization differentially. The original inoculant and dilutions will be cultured anaerobically to collect conditioned media (CM) enriched with microbial secretomes. CM will be used to expose a fluorescence resonance energy transfer (FRET) biosensor assay and a Gaussia luciferase protein complementation assay – both modified to quantify αSyn-αSyn interaction indicating oligomerization. Any CM-altering αSyn oligomerization will undergo multiomic characterization to identify potential causative agent(s). Results/Anticipated Results: Specific microbe-produced molecules from the literature are anticipated to modulate αSyn oligomerization, identified by targeted, reductionist studies that selected and tested separately single microbial factors on αSyn aggregation in vitro and in vivo. From these studies, lipopolysaccharide and bacterial amyloid protein are expected to increase αSyn oligomerization, while short-chain fatty acids, such as butyrate, are expected to interfere with and decrease oligomerization. As a complementary systemic approach, this study’s agnostic methods involving MSA stool culture combined with the proposed dilution-to-extinction method are expected to identify additional MSA stool secretome components modulating αSyn oligomerization that might otherwise be missed in earlier reductionist approaches. Discussion/Significance of Impact: Completion of this reverse-translational work will aid in discovering MSA stool secretome components modulating αSyn oligomerization. Identification of specific factors contributing to pathologic αSyn behavior might set the stage for patient screenings for identified stool markers and could lead to microbiome-based interventions for MSA.

Objectives/Goals: This research aims to harness electronic health records (EHR) combined with machine learning (ML) to predict necrotizing enterocolitis (NEC) in preterm infants using data up to their first 14 days of life. We aim to provide interpretable results for clinical decisions that can reduce infant mortality rates and complications from NEC. Methods/Study Population: Through a retrospective cohort study using data from the University of Florida Integrated Data Repository and One Florida, we will develop machine learning models suitable for sequential data to predict NEC. Our inclusion criteria include very low birth weight (VLBW; <1500g) infants born <32 weeks gestation and EHR data availability from the first 14 days of life. We will include infants with NEC and infants without NEC to train our ML model. Exclusion criteria include infants diagnosed with spontaneous intestinal perforation and severe congenital anomalies/defects requiring surgery. Results/Anticipated Results: We anticipate that our model will provide an accurate and dynamic prediction for the risk of NEC in neonates using data up to the first 14 days of life. Our model will be interpretable to identify key risk factors and can integrate real-world clinical insights to increase early detection and improve patient outcomes. Discussion/Significance of Impact: The development of a model to predict NEC could be used in neonatal intensive care guidelines and protocols and could ultimately decrease mortality, reduce complications, improve the overall quality of care, and lower healthcare costs associated with NEC.

Objectives/Goals: As the aging population increases, maintaining cognitive and physical health becomes crucial. Executive functions (EF), including reaction time, sustained attention, and spatial memory, are essential for daily life and independence in older adults but tend to decline with age, especially in Alzheimer’s disease (AD) patients. Methods/Study Population: Physical balance (PB) impairments further exacerbate this decline, affecting the quality of life and independence. Balance training (BT) emerges as a potential non-pharmacological intervention to enhance EF. This study aims to 1) assess the association between PB impairments and EF deterioration in older adults with AD; 2) evaluate the impact of BT on EF enhancement in this population; and 3) explore changes in brain-derived neurotrophic factor (BDNF) biomarkers before and after physical activity related to BT. This pilot study employs a cross-sectional design with older Hispanic adults (>65 years) in Puerto Rico, comparing a control group without BT intervention to an experimental group with BT intervention. Both groups will be assessed for EF and BDNF biomarkers pre- and post-physical activity. Results/Anticipated Results: Expected outcomes include identifying significant correlations between PB impairment and diminished EF, demonstrating measurable EF improvements following BT, and evidencing sustained BDNF release post-BT despite PB impairments. By understanding the biological mechanisms linking BT and cognitive improvements, particularly the role of BDNF, this research could inform future strategies to mitigate cognitive decline in AD patients through targeted physical interventions. Discussion/Significance of Impact: The findings of this study could provide valuable insights into the development of new preventive interventions, enhancing the quality of life and independence for older adults.

Objectives/Goals: Our goal is to develop a CRM platform to streamline support for investigators, optimize resource use, and enhance service delivery within a CTSA institution. This platform will measure outcomes, improve transparency, and ensure compliance, creating an efficient, supportive research environment. Methods/Study Population: Beginning with the design concepts and collecting technical requirements, ensuring alignment with the needs of investigators, administrators, and service units. Apply project management methodologies to iteratively design, prototype, and test features. Focus on user-centric interfaces and seamless integration with institutional systems. Incorporate stringent data security protocols in compliance with regulatory standards. Conduct pilot studies to refine functionalities and gather constructive feedback. Facilitate extensive user training programs and offer continuous support. Regularly monitor performance metrics, iterating based on empirical data and user feedback to ensure continuous enhancement and adherence to institutional objectives. Results/Anticipated Results: YCCI successfully designed, developed, and implemented a CRM platform for managing the intake, navigation, consultation, service delivery, and outcomes tracking of the clinical and translational science activities that are supported by and beyond the CTSA-supported infrastructure. The CRM platform has been live in production since roughly July 1, 2024, and YCCI is excited to share data, results, and outcomes with the ACTS community. Discussion/Significance of Impact: YCCI’s CRM platform advances the CTS mission by streamlining PI support, enhancing resource utilization, and fostering inter-team collaboration within the research ops and administration teams. Measuring outcomes and service efficiency enables data-driven decisions, ensuring continuous improvement, and impactful translational science.

Objectives/Goals: Congenital cytomegalovirus (cCMV) continues to be the primary infectious cause of fetal anomalies. The role of fetal natural killer (NK) cells in response to cCMV remains largely unexplored. This study seeks to investigate how fetal NK cells respond to human cytomegalovirus (HCMV) during gestation. Methods/Study Population: Umbilical cord blood and corresponding umbilical cord tissues were collected from fetuses that had no complications during gestation. These samples, provided by the Medical College of Wisconsin Tissue Bank, were processed within 24 hours after live birth. Single-cell suspensions were prepared from the samples, and fetal NK cells were isolated and exposed to HCMV antigen peptides VMAPRTLFL, VMAPRTLIL, VMAPQSLLL, and the human self-peptide ALALVRMLI. These peptides were presented on HLA-E*01:03 BV421-conjugated tetramers produced by the National Institutes of Health Tetramer Core Facility. Additionally, fetal NK cells were also prepared for single-cell RNA sequencing (scRNA-seq), and cells were filtered and clustered based on the number of uniquely expressed genes. Results/Anticipated Results: Through unbiased clustering, our scRNA-seq analysis identified five unique fetal NK cell subsets in umbilical cord blood and four in the corresponding umbilical cord tissue. Notably, fetal NK cells exposed to HCMV during gestation were primarily mature NK cell subsets, while those from unexposed fetuses were mostly immature subsets. Additionally, HCMV-exposed fetal NK cells exhibited a strong recall response to the HCMV antigen, with a notably higher frequency and elevated production of IFN-γ. Conversely, naïve fetal NK cells from fetuses unexposed to HCMV produced significantly lower levels of IFN-γ. Finally, we identified a distinct subset of fetal NK cells that emerge following exposure to the HCMV antigen. Discussion/Significance of Impact: In this study, we show that HCMV infection can influence the formation of specific NK cell subsets and re-exposure to the HCMV antigen can trigger a recall response. These insights could pave the way for the development of innovative NK cell-based immunotherapies aimed at preventing fetuses from developing symptomatic cCMV.

Objectives/Goals: Upon diagnosis, patients with acute myeloid leukemia (AML) have significant information needs. Given its recent increase in popularity, patients may use ChatGPT to access information about AML. We will examine the quality, reliability, and readability of information that ChatGPT provides in response to frequently asked questions (FAQs) about AML. Methods/Study Population: From FAQs on the top 3 patient-facing websites about AML, we derived 26 questions, written in lay terms, about AML diagnosis, treatment, prognosis, and functional impact. We queried ChatGPT-4o on 10/14/2024 using a new Google account with no prior history. We asked each question in a separate chat window once, verbatim, and without prompt engineering. After calibration, 5 oncologists independently reviewed ChatGPT responses. We assessed quality via the Global Quality Scale (GQS), scored from 1 (poor) to 5 (excellent) based on flow, topic coverage, and usefulness. For reliability, we assessed whether each response addresses the query and is factually accurate, elaborating on specific inaccuracies. For readability, we assessed Flesch-Kincaid Grade Level, Gunning Fog Index, and Simple Measure of Gobbledygook. Results/Anticipated Results: This will be a descriptive analysis of ChatGPT responses. For quality and reliability assessments, we will report Fleiss’ kappa for inter-rater reliability and expect substantial agreement or greater (≥0.61). Per prior studies in other domains, we hypothesize that ChatGPT responses will have good quality on average (i.e., GQS score near 4). We hypothesize that nearly all responses will address their query and will mostly be accurate; a minority of responses may have partial inaccuracies. Finally, we hypothesize that readability metrics will suggest that a higher educational level (e.g., college-level education) is required for comprehension. Overall, these findings will help elucidate strengths and limitations of ChatGPT for AML and guide discussion of factors patients should be aware of when using ChatGPT. Discussion/Significance of Impact: No prior study has examined the educational quality of ChatGPT for AML. Our study will detail whether patients are receiving trustworthy and meaningful information, identify misinformation, and provide guidance to oncologists when recommending information resources to patients or fielding questions that patients may raise after using ChatGPT.

Objectives/Goals: Diabetic kidney disease (DKD) affects 40% of diabetic patients, leading to renal failure, yet the molecular drivers remain elusive. MicroRNAs, noncoding regulators of gene expression, may hold the key. This study aims to identify key miRNAs in DKD, providing crucial insights for early intervention. Methods/Study Population: miRNA sequencing was conducted on kidneys from 8-week old male BTBR wild type and BTBR ob/ob mice. BTBR ob/ob mice lack the hormone leptin and spontaneously develop type 2 diabetes, with morphological renal lesions characteristic of human DKD. Total RNA was extracted from whole kidney sections and processed using the QIAseq miRNA library kit. Sequencing was performed on an Illumina NextSeq 550 platform. GeneGlobe analysis was used to identify differentially expressed miRNA functional pathways, while ingenuity pathway analysis (IPA) was employed to predict master regulators and causal networks involved in DKD. Results/Anticipated Results: miRNA sequencing identified significantly differentially expressed miRNAs (p < 0.05) between 8-week-old BTBR WT and BTBR ob/ob male mice, including miR-34a (-6.86 fold), miR-122 (-5.01 fold), miR-129 (-2.23 fold), miR-142a (+2.78 fold), miR-346 (+4.66 fold), miR-547 (-2.49 fold), miR-592 (+11.81 fold), miR-802 (-6.95 fold), and miR-6539 (-7.93 fold). Qiagen GeneGlobe analysis revealed biological processes potentially targeted by these miRNAs, including endocytosis, phagocytosis, hyperglycemia (p = 7.59e-3), and insulin-dependent diabetes (p = 4.32e-4). IPA predicted activation of RRAS, a small GTPase regulating cell growth and signaling (Z-score +2), with miR-34a and miR-122 targeting MYC, PI3K, and TGF-β in DKD progression in BTBR ob/ob mice. Discussion/Significance of Impact: We identified kidney miRNA expression in BTBR ob/ob mice at a pivotal disease stage. miR-34a, miR-122, and RRAS emerged as key drivers in DKD progression, showing remarkable early biomarker potential. These findings lay the groundwork for early detection and innovative therapies to halt DKD and improve patient outcomes.

Objectives/Goals: Clinical and Translational Science Award hubs will be the primary investors in advancing translational science until academic reward structures and funding agencies incentivize these efforts. As such, hubs will benefit from systematic methods to strategically identify and efficiently solve challenges in the translational process. Methods/Study Population: Translational science (TS) problems are abundant, complex and typically reside within systems of interconnected processes and people. These characteristics are informing efforts at the Michigan Institute for Clinical & Health Research (MICHR) to create a framework that guides how we select the right translational science problems to invest in solving and how we solve these problems in user-centric, efficient, and effective ways. Our framework leverages methods from the fields of design and systems thinking. Design thinking is a human-centered approach to problem solving and innovation that is ideal for tackling ill-defined and complex problems. Systems thinking methods help us situate and analyze problems within broader dynamics, structures, and perspectives, ultimately informing key levers for change. Results/Anticipated Results: Applying design and systems methods, we created an eight-step TS framework that centers on the diverse perspectives of those experiencing a TS problem and those implementing solutions. The first four steps guide in defining the TS problem within its context, understanding why previous solutions have not worked, and determining the value and generalizable knowledge that a solution would create. The last four steps are solutions-focused, with iterative brainstorming, testing, and refining of potential solutions before they are implemented locally and disseminated widely. Each step is underpinned by guiding questions, methods, outputs, and metrics to ensure a scientifically rigorous approach to defining, prioritizing, and solving TS problems. We are currently prototyping the framework with various case studies. Discussion/Significance of Impact: A framework that guides strategic investment in TS should ensure resources are allocated to the most well-defined and pressing problems that are important to the community and should speed up the process of creating solutions. Engaging myriad viewpoints leads to more viable solutions that foster a commitment to real change within the research ecosystem.

Objectives/Goals: We will conduct a 12-week pilot randomized controlled trial (RCT) to test the feasibility, acceptability, and preliminary efficacy of a staged-intensity whole foods intervention on hemoglobin A1c (HbA1c) change in adults, diet quality change (via the 2020 healthy eating index [HEI-2020]) in adults and offspring, and diet adherence and social determinants of health (SDOH) considerations via focus groups. Methods/Study Population: In this two-arm, parallel RCT, 30 adults with prediabetes (25–59 years) and their offspring (6–18 years) will be randomized to receive the 1) 12-week whole foods intervention which includes a 2-week feeding period (all foods/recipies provided), a 6-week customizable feeding period (3 dinners/recipies weekly), and a 4-week maintenance period (no food/recipies). The control group will receive standard of care (i.e., single RD-led diet counseling session). Primary outcomes include feasibility (≥80% retention and completion of study outcome measures) and acceptability (≥75% adult self-reported diet satisfaction). Intervention effects include 1) HbA1c change at 12-weeks in adults and 2) adult/offspring HEI-2020 scores assessed via diet records. Focus groups will assess influences of SDOH on diet adherence. Results/Anticipated Results: We have received Institutional Review Board approval, and recruitment is planned for January 2025. We will enroll 30 families from the greater Nashville, TN area. An intent-to-treat analysis will be conducted to test the preliminary effects of the whole foods diet intervention on the 12-week change in HbA1c (adults only) and 2020-HEI diet quality scores during the intervention period (adults and offspring). Focus groups will be conducted to understand how individual and family needs/preferences and SDOH may be perceived barriers or facilitators of diet adherence. Data generated from this study will be used to guide a fully powered RCT of our whole foods intervention to assess long-term effects on additional diabetes and metabolic outcomes and assessment of SDOH influences to support long-term adherence. Discussion/Significance of Impact: A healthy diet pattern is an effective nonpharmacological solution to prevent T2D, but only if it can be maintained. A family-centered whole foods diet pattern that uses “food as medicine” and considers how individual and family needs/preferences, and SDOHs could be an effective and sustainable multigenerational solution to prevent T2D in families.

Objectives/Goals: Using k-means clustering, we aimed to identify whether clusters characterized by greater exposure to minority stressors (e.g., experienced/anticipated discrimination) were associated with lower moderate-vigorous physical activity (MVPA) and higher sedentary behavior among sexual and gender minority (SGM; e.g., lesbian/gay, bisexual, transgender) adults. Methods/Study Population: SGM adults face higher risks of inadequate PA due to exposure to minority stressors, which may influence their willingness and ability to engage in PA. An online sample of SGM adults completed daily surveys about experienced and anticipated discrimination and wore wrist accelerometers for 28 days to objectively measure PA. We used k-means clustering to identify clusters based on reports of experienced and anticipated discrimination. We determined the optimal number of clusters using established partition criteria and ran linear regression models (adjusted for demographic factors) to examine the associations of minority stress clusters with MVPA and sedentary time. Results/Anticipated Results: Among 42 SGM adults (mean age 27.0±7.7 years) with 1133 person-days of accelerometry data, we identified four minority stress clusters: low anticipated/low experienced (LALE; reference group), low anticipated/high experienced (LAHE); high anticipated/low experienced (HALE), and high anticipated/high experienced discrimination (HAHE). Participants in the HALE cluster (n = 12) engaged in 202 fewer minutes of MVPA than those in the LALE cluster (n = 7). Participants in the LAHE cluster (n = 10) had 123 fewer minutes of vigorous PA than those in the LALE cluster. No differences were identified for sedentary time. Discussion/Significance of Impact: This is the first study to examine the association of minority stressors with objective PA among SGM adults. Participants in the HALE/LAHE clusters had lower PA than those with low levels of experienced and anticipated discrimination. Interventions to improve PA among SGM adults must target reduction of minority stressors.

Objectives/Goals: We aimed to discover treatment candidates for uterine fibroids, a common benign tumor with adverse impacts on quality of life. Repurposing already approved medications for fibroids can expedite treatment option expansion. Using genetic proxies, we identified novel fibroid drug candidates and estimated their effect on risk of fibroid diagnosis. Methods/Study Population: We performed a genetically predicted gene expression (GPGE) analysis using S-PrediXcan and GTEx tissue models with multi-ancestry genome-wide association study (GWAS) summary statistics of fibroids (cases  =  74,294, controls  =  465). There were 81 genes significantly associated with fibroid risk. Querying drug–gene interaction databases identified 56 approved medications that target these genes, including two antihypertensives, hydralazine, and spironolactone. Using independent multi-ancestry GWAS summary statistics (N  =  635,969) for systolic (SBP) and diastolic blood pressure (DBP), we conducted GPGE analyses. Blood pressure (exposure) and fibroids (outcome) GPGE summary statistics in the same tissues were used for two-sample Mendelian randomization (MR) analyses to proxy medication effects. Results/Anticipated Results: GPGE analyses identified hydralazine/tumor protein P53 (TP53) activity and spironolactone/thyroid hormone receptor beta (THRB) activity as drug-gene candidate pairs. Both drugs increase gene activity of their paired gene. Increased TP53 expression was associated with SBP in four tissues (exposure). The MR results indicated hydralazine use, proxied by increased TP53 expression, may reduce fibroid risk by 42% per standard deviation of gene expression (odds ratio [OR]  =  0.58, p  =  1.43E-12). Increased THRB expression was associated with DBP in eight tissues and were included in the MR (exposure). The MR results suggest spironolactone use, proxied by increased THRB expression, may reduce fibroid risk by 23% per standard deviation of gene expression (OR  =  0.77, p  = 5.94E-6). Discussion/Significance of Impact: We provide biologically plausible evidence for repurposing hydralazine and spironolactone for reducing risk of fibroid diagnosis. Repurposing these hypertension medications could provide novel preventative treatments for fibroids, particularly for individuals disproportionately affected by both conditions.

Objectives/Goals: The aim of the study is to identify resistance factors for substance use (i.e., factors that explicitly help to avoid or reduce drug use). Identification of resistance factors could inform strategies that seek to reduce the prevalence of substance use and related disorders. Methods/Study Population: Adult twins aged 30–70 years were recruited from the Mid-Atlantic Twin Registry. A mixed-method approach, group concept mapping, was used to identify factors influencing participants to resist using substances. Approximately 155 participants produced 97 statements reflecting substance use resistance factors. Hierarchical cluster analysis and multidimensional scaling assessed how participants sorted and rated statements for their lifetime and current importance. Factor analysis was used to reduce data dimensionality. Reliability analyses were conducted to identify a subset of statements anticipated to consistently represent each cluster. Results were shared with participants to assess accuracy with their experiences. Results/Anticipated Results: Participants sorted 97 statements into 9 thematic clusters: (1) Controlling Personal, Negative Consequences; (2) Concern About Health and Well-being; (3) Lack of Desire; (4) Outside Influences; (5) Social Norms and My Reputation; (6) Career and Legal Impacts, (7) Avoiding Harm to Family and Relationships; (8) Preserving Family Relationships; and (9) Family and Friends Impact on Me. Participants consistently identified health concerns as an important substance use resistance factor. The statements will be further reduced to represent a smaller subset for future use as a scale to measure exposure to resistance factors. Discussion/Significance of Impact: Health concerns related to substance use were identified as an important resistance factor. This has been supported by research on smoking cessation and implemented in smoking prevention campaigns. Therefore, prioritizing health-related outcomes in prevention may be important to reduce substance use prevalence.

Objectives/Goals: We aim to identify how IDH mutant (IDHm) gliomas use exosomes to modulate the local and systemic immune system. We will do so by characterizing differential miRNA expression between IDHm and IDH wild type (IDHwt) exosomes and identifying the specific immune cell population targeted by exosomes in vivo. Methods/Study Population: Exosome RNA will be isolated from cultured patient glioma samples and perform small RNA sequencing to investigate differential expression of miRNA between IDHwt and IDHm exosomes. We will then utilize miRNA target databases in conjunction with bioinformatic pathway analysis to generate potential target regulatory pathways. To identify the in vivo effect of tumor exosomes, we will generate a novel glioma mouse model that has been genetically engineered to release labeled exosomes using the RCAS retroviral system. We will collect peripheral blood and tumor tissue for flow cytometric immune profiling and single-cell RNA sequencing. The transcriptomic data will be analyzed to identify subsets of immune populations that have taken up the labeled exosomes and assess the resulting expression changes in those cells. Results/Anticipated Results: From the small RNA sequencing and bioinformatics analysis, we expect to find several unique miRNA expressed in IDHm exosomes that induce immunosuppressive pathways in local and systemic immune cell populations when compared to IDHwt exosomes. Furthermore, using our novel murine model, we expect to be able to track endogenously released exosomes in the local tumor microenvironment and in the circulating blood. We hypothesize that IDHm exosomes specifically target precursor myeloid cells within the local and peripheral circulating immune populations and induce the expansion of monocytes, M2 macrophages, and mono-MDSCs. Discussion/Significance of Impact: Immunosuppression in IDHm glioma has hindered the development of adequate therapies to treat this fatal disease. Our study will illuminate the mechanism by which tumor exosomes can suppress immune surveillance. These results will help identify new therapeutic targets to sensitize the immune system against glioma cells.