Objectives/Goals: We hypothesized that adding cytomegalovirus (CMV) viral Fc gamma receptors (vFcγRs) to a glycoprotein B (gB) protein subunit vaccine would improve vaccine-elicited Fc mediated effector functions such as antibody dependent cellular phagocytosis (ADCP) and cytotoxicity (ADCC), over gB subunit alone. Methods/Study Population: We immunized rabbits (n = 4 per group) at Weeks 0, 4, and 8 with 20μg gB alone or with one vFcgR (gp34, gp68, or gp95) at 20μg or 40μg, adjuvanted with squalene emusion, Addavax. Plasma from immunized rabbits was analyzed for antigen-specific IgG binding via enzyme-linked immunosorbent assays (ELISAs). ADCP was measured by conjugating whole virions to a fluorescent marker (AF647), incubating the fluorescent virus with rabbit plasma, and measuring uptake of virus by THP-1 monocytes via flow cytometry. ADCC was measured by natural killer cell degranulation via flow cytometric detection of CD107a expression following co-incubation with CMV-infected fibroblasts and rabbit plasma. Results/Anticipated Results: Each vFcγR demonstrated immunogenicity, although average vFcγR-binding IgG titers were between 4- to 10-fold higher in animals receiving the 40μg dose of each vFcγR compared to the 20μg dose. We observed similar IgG binding responses against gB among all vaccine groups. Comparing groups at peak immunogenicity (Week 10), ADCP responses were improved over gB alone by approximately twofold in animals receiving 40ug of each vFcγR. This effect was maintained across several human CMV strains with variable vFcγR genes. ADCC responses were undetectable in all animals immunized with gB alone, yet those receiving 40μg gp34 or gp95 demonstrated detectable ADCC. Discussion/Significance of Impact: HCMV-specific ADCP and ADCC are associated with protection against vertical CMV transmission, so a vaccine including vFcγRs which can improve vaccine-elicited Fc-effector responses is promising toward reducing the immense global impact of congenital CMV and associated neurologic birth defects.

Objectives/Goals: This study aims to evaluate diversity of participants in GLP-1 T2DKM clinical research with regard to sex, race, and ethnicity by using results data available through the ClinicalTrials.gov database. Sample population estimates for studies were calculated using the 2020 Census and compared within groups with respect to sex, race, and ethnicity. Methods/Study Population: The public ClinicalTrials.gov database was searched for interventional studies with GLP1 inclusion as treatment (n = 2,397). This search was then filtered to studies where results were reported (n = 772). From these studies, 466 studies focused on type 2 diabetes as a condition and thus became the analysis dataset. Participant and protocol information for these 466 studies were obtained from the clinical trials transformation initiative (CTTI) as an AACT data download. Observed to expected ratios were calculated for each subgroup-based population estimates from the 2020 Census and using the baseline counts of participants for studies where sex, race, and ethnicity were provided. In addition to within group comparisons, study characteristics (e.g. phase) were included in models to assess influence of covariates. Results/Anticipated Results: Of the 466 studies, 430 (92%) reported sex, 171 (37%) reported race, and 145 (31%) reported Hispanic ethnicity. Among those found to be underrepresented in studies (defined as a ratio < 1): females (mean  =  0.89, median  =  0.92); Black/African Americans (mean  =  0.88, median  =  0.39). Hispanic or Latinos mean ratio was 1.16 (95% CL: 0.97, 1.35) but had the least available data. When including covariates in the models, there were statistically significant differences in ratios with respect to sex as females had significantly lower odds compared to males (ratio > =  1), with the odds being about 21% of those for males. With respect to race, Black or African American individuals had significantly lower odds (about 32% of those of White individuals) (ratio > =  1). Discussion/Significance of Impact: This study reveals significant underrepresentation of females (mean ratio 0.89) and Black/African Americans (mean ratio 0.88) in clinical trials for GLP-1 drugs in type 2 diabetes. These disparities highlight the need for more inclusive research to ensure diverse populations benefit equally from medical advancements.

Objectives/Goals: Collaborations between translational science programs and academic health sciences libraries can enhance research impact by improving efficiency, leveraging diverse professional expertise, and expanding opportunities for collaboration between librarians and translational science programs. Methods/Study Population: A team science approach was utilized, integrating findings from a literature review, practical experiences of health sciences librarians, and collaborative writing. An analysis of case studies from institutions with successful partnerships explored the roles of libraries in partnering with translational science programs. The data collected were mapped to the Clinical and Translational Science Award Program’s five functional areas outlined in the Notice of Funding Opportunity PAR-24–272. Librarians from 21 institutions engaged in discussions and collaborative writing to share insights and identify key factors driving successful partnerships. Results/Anticipated Results: Academic health sciences libraries play a crucial role in enhancing translational science programs through expert knowledge management, facilitation of research dissemination, and support for interdisciplinary collaboration. Results from this project include a table outlining 16 specific opportunities mapped across five functional areas and six topical categories for translational science programs and libraries to collaborate effectively.Successful partnerships demonstrate improved research workflows, increased interactions between researchers and libraries, and accelerated translation of discoveries into clinical settings. These collaborations illustrate opportunities for other institutions to adopt as they consider best practices in supporting translational science. Discussion/Significance of Impact: By combining resources and expertise between libraries and translational science programs, these partnerships enhance the ability to transform scientific discoveries into real-world clinical applications, drive innovation, and amplify the contributions of both libraries and translational science programs.

Objectives/Goals: Community engagement is critical to promoting equity for those affected by clinical and translational research. Despite its importance, investigators rarely involve community stakeholders in ancillary projects such as secondary analyses. I use a study example to demonstrate how to meaningfully partner with communities in ancillary research. Methods/Study Population: Using data collected in a previous study, the aim of this secondary analysis was to examine whether pregnant women’s reports of social adversity during the third trimester were associated with their infant’s resting cortisol level at 1, 6, and 12 months of age. Community engagement at the study design phase included consultation with an institutional Community Advisory Board (CAB) whose expertise included lived experiences relevant to the context of the study (i.e., social adversity and pregnancy). Community engagement at the analysis and dissemination stage included deeper collaboration with three community research consultants (CRCs) from the CAB; all identified as women of color and mothers with relevant lived experiences. CRCs received compensation for time and effort and an invitation for co-authorship. Results/Anticipated Results: Consultations included written review as well as a live presentation and discussion with the CAB for feedback on the study proposal prior to submission for IRB approval. The focus of discussion was around the use of inclusive terminology and ensuring respectful and equitable representation of all participants in the original study, including those who reported experiences with social adversity during pregnancy. After receiving study approval, statistical analyses were finalized and CRCs actively engaged in discussions with the PI around the interpretation of results. CRCs significantly contributed to the development of solutions-oriented implications for practice, policy, and future research. All CRCs were co-authors on the study manuscript that was published in a top-tier international journal. Discussion/Significance of Impact: Community engagement in ancillary research is feasible and essential. True equitable collaboration requires partnership during all study activities, including critical phases (e.g., developing aims, data analysis). Researchers also should prioritize the use of datasets from studies that incorporated community partnership and engagement.

Objectives/Goals: 1) Determine the association between wildfire smoke-driven PM2.5 and risk of persistent respiratory symptoms and repeated asthma exacerbations after the acute wildfire period among adults with asthma. 2) Examine how measures to reduce personal exposure to wildfire smoke, including avoiding outdoor activities, modify this association. Methods/Study Population: This is a retrospective study of adults with asthma in WHAT-NOW, a cohort study of people living in Northern California during the 2018 Camp Fire. Daily smoke-driven PM2.5 was estimated for each participant based on their home address or evacuation location. We examined the association between mean PM2.5 exposure and the presence of respiratory symptoms at both the time of the survey (6–16 months post-wildfire) and at least one other post-wildfire time-period, as well as whether they had a medically attended respiratory illness (saw a doctor, visited the ER, or were hospitalized for a respiratory symptom). We examined the interaction of PM2.5 with spending time outdoors during the wildfires. Poisson regression models with robust standard errors were adjusted for age, sex, race, smoking, allergies, and education. Results/Anticipated Results: Among 337 adults with asthma in the WHAT-NOW cohort, one standard deviation higher smoke-driven PM2.5 was associated with higher risk of any persistent respiratory symptom (risk ratio (RR) 1.38, 95% CI 1.07 – 1.78) and having at least one medically attended respiratory illness (RR 1.33, 95% CI 1.07 – 1.65), but not significantly associated with repeated asthma exacerbations (RR 1.30, 95% CI 0.92 – 1.81). However, there was a significant interaction between PM2.5 and outdoor activities during the wildfire on the outcomes of any persistent respiratory symptoms (p = 0.041) and repeated asthma exacerbations (p = 0.028). The association between PM2.5 and repeated asthma exacerbations was greater among people who spent time outdoors (RR 3.36, 95% CI 1.47 – 10.23) than those who did not (RR 1.00, p = 0.99). Discussion/Significance of Impact: This study provides evidence that exposure to wildfire smoke increases respiratory morbidity among adults with asthma beyond the acute wildfire period. Additionally, it suggests that avoiding outdoor activities on smoky days can significantly decrease the risk of future repeated asthma exacerbations associated with smoke exposure.

Objectives/Goals: The extracellular matrix (ECM) of the sinoatrial node (SAN) is critical for maintaining automaticity in hiPSC-derived pacemaking cardiomyocytes (PCMs) under cyclic strain. We aim to determine the ECM ligands responsible for cell-ECM mediated mechanotransduction and the resulting phenotype in PCMs. Methods/Study Population: HiPSCs are differentiated to PCM and replated on substrate with 5 or 15 kPa PDMS that are coated with 5 or 25 ug/cm of either collagen I or fibronectin at sub-confluent density to restrict junction engagement to only costameres. Then, PCM are subjected to 10% cyclic mechanical strain at 1 Hz for 48 hours, with static culture as control. PCMs from all conditions are subsequently fixed and stained for cardiomyocyte-specific troponin T (TnT), pacemaking HCN4 channel, and pro-pacemaking transcription factors (Shox2, Isl1, Tbx3, Tbx18). Additionally, PCM cell size will also be assessed. Results/Anticipated Results: Considering the amount of hypertrophy and myofilament in CMs correlates with mechanical strain, we expect a reduced degree of mechanotransduction in hiPSC-PCM on collagen I with a stiffness 15 kPa to induce smaller cell size with fewer myofilament and an upregulation of HCN4 and pro-pacemaking transcription factors than those on 5 kPa and those on fibronectin of either 5 or 15 kPa after cyclic strain. This is because COL1 is reported to have a lower signaling threshold but a limited sensitivity to force which contributes to the diminished mechanotransduction signaling. Discussion/Significance of Impact: Effects of the microenvironment on hiPSC-PCMs via costamere mechanotransduction may provide insights for engineering biopacemakers with a suitable ECM, to potentially preserve automaticity in hiPSC-PCMs and sustain long-term pacemaking function, making biopacemakers a step closer to reality.

Objectives/Goals: This study aims to uncover immuno-endotypes in sepsis-associated pediatric acute respiratory distress syndrome (SA-PARDS) by using single-cell RNA sequencing (scRNA seq) to analyze the immune cell populations of the lower respiratory tract of intubated pediatric subjects with SA-PARDS. Methods/Study Population: Inclusion criteria are age less than 18 years, admission to the PICU, diagnosis of SA-PARDS, and intubation. Both sepsis and PARDS will be defined using the most recent consensus definitions. Exclusion criteria include an order of limited resuscitation and clinician discretion. After informed consent is obtained, a tracheal aspirate and blood sample will be obtained on days 1, 3, and 7. Both samples will be processed for single-cell RNA seq via the HIVE platform per manufacturer protocol. cDNA libraries will then be sent for 150 base pair paired-end sequencing. Sequences will be aligned to a reference genome, and count matrices will be generated. The Seurat package in R will be used for cell-type annotation and analysis of differential gene expression. Clinical variables, labs, and outcomes will be recorded in REDCap. Results/Anticipated Results: We expect to find that subjects with nonresolving SA-PARDS, defined as intubation and mechanical ventilation, will have a monocyte/macrophage transcriptome characterized by continued hyper-inflammation (M1-like phenotype) that does not transition over time to an anti-inflammatory and pro-repair phenotype (M2-like). Additionally, we expect to see that subjects with non-resolving SA-PARDS will have evidence of continued inflammation driven by hyper-inflammatory neutrophils. Finally, we expect that subjects with non-resolving SA-PARDS will have epithelial cells characterized by continued upregulation of canonical pathways of innate immunity including interferon signaling and the damage associated molecular pattern recognition pathway. Discussion/Significance of Impact: The discovery of immuno-endotypes in SA-PARDS would represent a major step toward developing precision medicine therapies for this group of patients. It would simultaneously provide a strategy to reduce biological heterogeneity and identify novel pathways and targets for therapy.

Objectives/Goals: REDCap is a popular electronic data capture tool. However, training users in how to best utilize REDCap can be a challenge for many institutions. The Clinical and Translational Science Institute (CTSI) strives to setup a self-service training program that takes the day to day roles of users into account. Methods/Study Population: Our new training curriculum is a collaboration with our Workforce Development team and our REDCap Support Team. The REDCap team functions as the subject matter experts and generate a training outline based on a certain feature or topic. The Workforce Development team transform that outline into an LMS style course that’s available online. In order to organize the courses for maximum relevance to user, we engaged with various REDCap training and regulatory experts around the globe. Based on their input, we organized the various training courses into a role-based schema. The training courses are freely available online and contain an optional test and completion certification in order to comply with regulatory standards like 21 CRF part 11 or GDPR. Results/Anticipated Results: We released the first 17 training courses in July 2024 with another 20 courses planned in the near future. Responses to the courses have been overwhelmingly positive from users and the greater REDCap community. Our collection of training courses won the best website award at the yearly REDCap conference in 2024. To date we have had 137 people go through a training course with the optional test and completion certificate. While the majority have been from the USA, a significant portion hails from other countries. We believed these people only represent a small subset of users due to the optional nature of the test and accreditation section. Discussion/Significance of Impact: Our new role-based training curriculum is crucial in giving REDCap users the training tools they need for their particular role. The certification option fills a niche for professionals to demonstrate their REDCap proficiency to further their careers. Overall, this user training should increase the utilization of REDCap in all research endeavors.

Objectives/Goals: Hearing loss (HL) can result from environmental and genetic factors. Some genetic variants may be more prevalent in populations living in geographic or cultural isolation. This study explores the genetic variants associated with HL in Puerto Rico and correlates these with auditory and balance disorders to uncover novel variants. Methods/Study Population: After obtaining individual informed consent and assent for a minor when applicable, we will collect clinical audiological data and biological samples (n  =  600) from families across Puerto Rico with a history of severe to profound HL. Genomic DNA will be extracted, and exome and mitochondrial genome sequencing will be conducted to identify causal variants in genes associated with HL. The study will assess the prevalence of both novel and reported variants in genes associated with HL and investigate founder variants in the Puerto Rican population. Involvement of genes so far not associated with HL will also be considered when a genetic diagnosis cannot be established. Auditory phenotypes will be correlated with genetic findings, allowing for a comprehensive analysis of genetic contributions to HL in this population. Results/Anticipated Results: This research will advance understanding of the genetic causes of HL in Puerto Rico, leading to more accurate diagnoses, personalized treatment options, and the discovery of novel genes associated with HL. It will also serve as an evidence-based reference to analyze the adequacy of current neonatal hearing screening protocols in PR. Recruitment and sample collection have begun, and we expect our findings to uncover population-specific variants. These results will provide a foundation for further genetic studies aiming at identifying the causes of HL in Puerto Ricans regardless of age of onset. Discussion/Significance of Impact: This study will enhance our understanding of hereditary HL and serve as a basis for developing population-specific diagnostic tools and interventions, particularly in the Puerto Rican population. The research will support future genetic studies and address health disparities in HL in the island.

Objectives/Goals: Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system that affects 2 million people worldwide causing severe disability. This study uses the Gamt-GFP transgenic mouse line as a novel approach to track oligodendrocyte lineage cell regeneration in inflammatory demyelination for identifying potential therapies. Methods/Study Population: We previously showed that Gamt, an enzyme required for creatine synthesis, is essential for oligodendrocyte (OL) maturation and survival using the Gamt-Green Fluorescent Protein (Gamt-GFP) reporter line. In this study, we capitalize on this finding and track OL lineage cells in an experimental autoimmune encephalomyelitis (EAE) mouse model by inducing immune-mediated demyelination in the Gamt-GFP reporter line. At 7 days postimmunization (dpi), both control and EAE mice receive 4 mg/kg tamoxifen for 4 consecutive days to induce GFP expression. GFP+ cells and those also expressing OL lineage markers [Olig2 (pan-OL lineage cell marker), NG2 (OL precursor cells; OPCs), and CC1 (mature OL)] are quantitated by immunofluorescent staining of spinal cord sections collected at 28 dpi. Results/Anticipated Results: Preliminary data using immunofluorescent staining demonstrated GFP was expressed in Olig2+ cells in the inflammatory ventral white matter lesions of mice with EAE, whereas no GFP labeling was present in the control mice. Moreover, GFP+ cells also expressed NG2+. In contrast, few CC1+ cells were detected in the inflammatory lesions. The low number of dual labeled GFP+CC1+ cells in these lesions suggests OPCs under the EAE environment are unable to efficiently differentiate into mature OL. Therefore, the Gamt-GFP reporter mouse can be used to identify and track activated OL lineage cell populations (i.e., GFP+CC1) in inflammatory lesions in the EAE mouse model. Discussion/Significance of Impact: The Gamt-GFP reporter line identifies activated OL lineage cells responding to inflammatory demyelination, making it a valuable tool for testing potential therapeutics aimed at enhancing remyelination. This model helps bridge the gap between preclinical and clinical research to guide MS therapy development.

Objectives/Goals: This study aims to explore the relationship between plasma biomarkers (GFAP, NF-L, and IL-1β) and cognitive impairment in moderate to severe TBI patients. We will assess biomarker levels and their link to neurocognitive outcomes at acute and chronic stages of injury. Methods/Study Population: We will recruit 100 patients aged 21 years and older with moderate to severe TBI (Glasgow Coma Score 3–12) from a trauma hospital. Blood samples will be collected at 24–72 hours post-injury and again at 3 and 6 months. Plasma levels of GFAP, NF-L, and IL-1β will be measured using multiplex ELISA. Neurocognitive tests will be administered at 3 and 6 months to assess cognitive function. Correlations will be made between biomarker levels, neurocognitive performance, and disability scores (Disability Rating Scale and Glasgow Outcome Scale). Exosome isolation from plasma will allow for detailed analysis of astrocyte-derived biomarkers and their association with long-term cognitive impairment and recovery. Results/Anticipated Results: We anticipate that plasma levels of GFAP, NF-L, and IL-1β will be elevated in the acute phase of moderate to severe TBI and will correlate with injury severity. At 3 and 6 months, higher levels of IL-1β, in particular, are expected to be strongly associated with cognitive deficits. We also anticipate that biomarkers in astrocyte-derived exosomes will provide more specific insights into long-term neuroinflammation and its impact on cognitive function. These findings could pave the way for targeted, personalized interventions to improve recovery in TBI patients. Discussion/Significance of Impact: This research focuses on inflammation’s role in cognitive impairment and disability in TBI patients. We propose using multiple biomarkers – GFAP, IL-1β, NF-L – paired with advanced techniques like exosomes and multiplex analyses to identify novel therapeutic targets, aiming for personalized treatment strategies, as well as prognosis.

Objectives/Goals: To design a flexible, comprehensive framework for Data Science Units to cultivate sustainable, long-term relationships with Clinical and Translational Science Research Units. Best practices for managing Data Science collaborations are presented to improve the quality and efficiency of research conducted throughout academic health centers. Methods/Study Population: Leaders of Data Science Units across six institutions formed a workgroup to develop guidance and best practices for Data Science Units to establish long-term, sustainable collaborations with Clinical and Translational Science Research Units. This guidance is based on tools and protocols developed and employed by the participating units, which range from larger groups with over 20 partnerships to a unit with three partnerships that is actively working to expand. Importantly, partnerships are highly variable, with some partnerships at one institution representing engagement with over 500 faculty, whereas some partnerships at another institution involve the lab of a single investigator. Results/Anticipated Results: We offer guidance in three domains: (1) Identifying the needs for a new partnership, including assessing required effort and data science expertise, setting partnership priorities, developing formal agreements, and identifying goals and metrics; (2) managing data science teams by implementing regular meetings, creating project intake and prioritization processes, and effort monitoring; and (3) evaluating the successes and failures/gaps of the collaboration by measuring the metrics mapped to the goals. For each domain, we provide specific suggestions on which parties should be involved and how frequently the processes should occur. This guidance is applicable both to larger collaborative partnerships and to smaller, single faculty or staff partnerships, whether they are new or well-established. Discussion/Significance of Impact: Effective collaboration between data scientists and clinical and translational investigators is key to advancing data-driven research. The guidance and resources are presented to support Data Science Units in successfully managing long-term collaborations through goal-development, evaluation, and adapting to evolving research needs.

Objectives/Goals: Community gardening can foster healthy behaviors among low-income communities. This project aimed to develop a community garden. The primary objectives of this project are (1) assessing the need for and perspective on a community garden at the childcare center, (2) installing the garden, and (3) engaging children in gardening education. Methods/Study Population: This project took place at a childcare center in Harrisburg, PA. Most (74.6%) residents identified as Black or Hispanic/Latino. Every child at the center was eligible for free or reduced lunch. A listening session was held with directly impacted community members to discuss the need for a community garden. Four caregivers, 1 early childhood educator and a master gardener (n = 6) attended the listening session, in which they shared their personal strengths and challenges in growing food. Attendees provided suggestions on what foods they wanted to grow. Children enrolled in the center’s summer program (n = 50) were then invited to participate in weekly gardening activities for 9 weeks. Activities were targeted to preschoolers (3- to 5-year-olds). Older children enrolled in the summer program were welcome to participate. Results/Anticipated Results: Feedback from the listening session was positive. Attendees provided ideas on what to grow and shared interest in expanding the garden to the broader community. Project staff installed four garden beds and planted a variety of herbs (basil, mint, and lavender), fruits (strawberry and melon), and vegetables (tomato, squash, pepper, and onion). Roughly 20–50 children were engaged in the garden each week. Eight weeks into the project, one member from the broader community noticed the garden’s growth and expressed gratitude to the staff, stating “I saw you when you first started planting. This is great what you are doing for the kids.” Children and the center’s staff responded positively to the activities. The staff expressed verbal gratitude for the project and were enthusiastic about maintaining the garden. Discussion/Significance of Impact: Developing a community garden was feasible in this sample and shows potential to (1) increase children’s food literacy and vegetable acceptance and (2) bridge the gap from farm to early childcare education. The project’s success paves way for future gardening initiatives that address food access issues within other diverse low-income populations.

Objectives/Goals: Adults from minority groups report more severe and pervasive pain than those in majority groups, resulting in a disproportionate burden of pain. Whether race disparities in pain outcomes exist in persons with multiple sclerosis (MS) is unknown. We examined the association of race with pain intensity, pain interference, and pain phenotypes in MS. Methods/Study Population: Ambulatory adults with medically documented MS completed a comprehensive survey battery including demographics and clinical data. Pain outcomes were assessed with four measures: Patient Reported Outcome Measurement Information System (PROMIS) pain intensity and pain interference short forms, the American College of Rheumatology Fibromyalgia Survey Criteria (a surrogate of degree of nociplastic pain), and the PainDETECT (a surrogate of neuropathic pain). Participants were categorized as either Black/African American or White based on their self-reported race. Four sets of unadjusted and adjusted (including sex, age, years since diagnosis, MS subtype and Patient Determined Disease Steps—PDDS score) linear regression models were built to examine the associations between race and pain outcomes. Results/Anticipated Results: A total of 258 participants (200 White and 58 Black), with a mean age of 51 ± 12 years, mostly female (77%), an average of 15 ± 10 years since diagnosis, a PDDS score ranging from 0 to 6, and mostly diagnosed with RRMS (79%), were included in the analyses. Unadjusted regression models indicated that pain intensity (β  =  5.20; 95% CI 2.73 – 7.66, p < 0.001), pain interference (β  =  5.17; 95% CI 2.29 – 8.06, p < 0.001), and nociplastic pain (β  =  2.41; 95% CI 0.40 – 4.42, p  =  0.019) were all higher for Black/African American participants compared to White participants. The differences remained statistically significant in adjusted models. No differences in neuropathic pain were observed between Black/African American and White participants in both unadjusted and adjusted models. Discussion/Significance of Impact: We highlight an increased burden of pain in Black/African American with MS compared with their White counterparts. The findings illuminate potential future targets of interventions to reduce disparities in the experience and impact of pain. A comprehensive examination of the role of social determinants in pain outcomes in MS is further warranted.

Objectives/Goals: Objectives/Goals (300 characters): Sarcopenia is a progressive skeletal muscle disorder associated with adverse outcomes. There is a gab of having objective measures upon performing interventions in patients with muscular dystrophies. The object of the present study is to describe the severity of sarcopenia in DMD patients in Puerto Rico. Methods/Study Population: Methods/Study Population (700 characters): Forty to 30 patients with DMD who are followed in MDA Care Center in the “Instituto De Rehabilitacion del Caribe.” Diagnosis will be confirmed with genetic testing and/or muscle biopsy. Lean muscle mass will be measured with a Whole Body Dexa (WBD) in a Nuclear Medicine Lab. Hand grip, elbow flexor, and knee extensor muscles strength will be measures with an isometric dynamometer. Patients’ functionality will done using the North Star Ambulatory Assessment scale and Brook and Vignos scales, which have been validated for patients with DMD and neuromuscular disease respectively. Correlations will be made with lean body mass (independent variable) and muscle strength and functionality (dependent variable). Results/Anticipated Results: Results/Anticipated Results (700 characters): We expect to find severe sarcopenia in patients with DMD in PR and that it will be more severe with older age. There will be a direct correlation between lean muscle mass and muscle strength, and functionality in DMD patients. Discussion/Significance of Impact: Discussion/Significance of Impact (300 characters): The findings of our study can help us to explore the possibility that Whole Body DEXA can serve as a potential biomarker for future studies since there is a need to develop noninvasive biomarkers that correlate with disease progression and interventions in DMD patients.

Objectives/Goals: Primary sclerosing cholangitis (PSC) manifests with an inflammatory milieu that leads to fibrotic scarring of the liver. Human PSC liver bile ducts are enriched with neutrophils; however, their infiltration and functional role is unexplored. Our goal is to investigate the mechanism and impact of peribiliary neutrophil infiltration observed in PSC. Methods/Study Population: Primary cholangiocytes (bile duct cells) isolated from WT and mouse models of PSC (3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-fed mice and Mdr2-/- mice) were analyzed by RNA-sequencing. Immunofluorescence (IF) was performed on liver tissues from PSC patients and mouse models of PSC for markers of bile ducts and neutrophils (KRT19 and MPO). Intrahepatic leukocytes (IHL) isolated from mice livers were evaluated for neutrophil abundance and activation state. Anti-Ly6G antibody-mediated neutrophil depletion in Mdr2-/- mice was analyzed by IF, histology, and cytometry by time-of-flight (CyTOF). Cholangiocytes stimulated with TNFα (to induce an inflammatory phenotype) were analyzed for neutrophil chemoattractants with genetic and pharmacological interventions. Results/Anticipated Results: RNA-seq analysis of primary cholangiocytes from PSC mouse models revealed enrichment in inflammatory and neutrophil degranulation pathways. Flow cytometry and RT-PCR analysis revealed an increase in the neutrophil population in PSC mice with activated phenotype. Peripheral depletion of neutrophils in Mdr2-/- mice alleviated liver injury and inflammation, along with a reduction in peribiliary neutrophil infiltration and attenuated bridging fibrosis. CyTOF analysis showed a significant reduction in CD8+ T cells upon neutrophil depletion, implying neutrophils sustain CD8+ T cells in PSC liver. Mechanistically in cholangiocytes, TNFα mediates expression of neutrophil chemoattractants, CXCL1 and CXCL8, through the cyclic GMP-AMP synthase stimulator of interferon genes (cGAS-STING) pathway. Discussion/Significance of Impact: Our findings suggest that activation of the STING pathway in cholangiocytes in cholestatic liver disease triggers an immune response resulting in peri-portal neutrophil infiltration via CXC chemokines. The sustained presence of these activated neutrophils engages the adaptive immune system to perpetuate the inflammation and fibrosis seen in PSC.

Objectives/Goals: Lung transplant is a life-saving surgery for patients with advanced lung diseases yet long-term survival remains poor. The clinical features and lung injury patterns of lung transplant recipients who die early versus those who survive longer term remain undefined. Here, we use cell-free DNA and rejection parameters to help elucidate this further. Methods/Study Population: Lung transplant candidacy prioritizes patients who have a high mortality risk within 2 years and will likely survive beyond 5 years. We stratified patients who died within 2 years of transplant as early death (n = 50) and those who survived past 5 years as long-term survivors (n = 53). Lung transplant recipients had serial blood collected as part of two prospective cohort studies. Cell-free DNA (cfDNA) was quantified using relative (% donor-derived cfDNA {%ddcfDNA}) and absolute (nuclear-derived {n-cfDNA}, mitochondrial-derived {mt-cfDNA}) measurements. As part of routine posttransplant clinical care, all patients underwent pulmonary function testing (PFT), surveillance bronchoscopy with bronchoalveolar lavage (BAL), transbronchial biopsy (TBBx), and donor-specific antibody testing (DSA). Results/Anticipated Results: Over the first 2 years after transplant, the number of episodes of antibody-mediated rejection (p) Discussion/Significance of Impact: Clinically, early-death patients perform worse on routine surveillance PFTs and experience a worse degree of CLAD. These patients also have higher levels of cfDNA as quantified by n-cfDNA and mt-cfDNA. These results provide preliminary evidence that early-death patients have worse allograft rejection, dysfunction, and molecular injury.

Objectives/Goals: This study aim to identify cervical cancer prevention barriers and facilitators, assess the associations between social determinants of health factors and cervical cancer prevention, and examine the association between levels of health literacy and willingness to undertake HPV vaccine and cervical cancer screening among Black and Hispanic women. Methods/Study Population: To achieve Aim 1, we will conduct a quantitative survey on barriers and facilitators to cancer prevention among non-Hispanic Black and Hispanic women. For Aim 2, we will assess the relationship between Social Determinants of Health and HPV vaccination/cervical cancer screening using the AHC Health-Related Social Needs Screening Tool. For Aim 3, health literacy will be measured using the SAHL–S&E test and its association with willingness to vaccinate or cervical cancer screening. We will analyze the results using chi-square and logistic regression models. Participants will be recruited through multiple methods. We will recruit 250 individuals who were assigned female at birth and identify as non-Hispanic Black or Hispanic, aged 18–26 years, from Caddo and Bossier Parishes. Results/Anticipated Results: We expect to identify several barriers and facilitators to cervical cancer prevention among non-Hispanic Black and Hispanic women, including factors like access to care, cultural beliefs, and knowledge gaps. Social determinants of health (SDOH), such as income, education, and healthcare access, will likely show a strong association with lower HPV vaccination, Pap smear, and HPV screening rates. Additionally, we anticipate that lower health literacy will correlate with reduced willingness to vaccinate or screen for HPV. These findings can bridge the gap between research and practical health applications by guiding the design of community-based behavioral interventions that enhance cervical cancer prevention among minority populations. Discussion/Significance of Impact: This research is the first to assess how SDOH factors impact cervical cancer prevention among Blacks and Hispanic women in North Louisianawhile also exploring the role of health literacy in HPV vaccination and screening. Findings will drive practical interventions to reduce disparities and improve outcomes.

Objectives/Goals: Physical therapy (PT) is a recognized and evidence-based component of oncology care that has been shown to benefit people with various cancers, such as breast, lung, head and neck, thyroid, or prostate cancer. The goal of this evaluation was to determine the level of PT service utilization by World Trade Center (WTC) Health Program members with cancer. Methods/Study Population: The Program is a limited benefits federal program that serves responders and survivors of the September 11th attacks in New York City, the Pentagon, and Shanksville PA. Our analyses include enrolled Program members with a cancer certification. Cancer types were divided into two categories, Category A (breast, lung, head and neck, thyroid, or prostate cancer) and Non-Category A (all other cancer types). Data included medical claims, certification, and enrollment data from July 2011 to December 2023. The 2023 Current Procedural Terminology (CPT) code list from the Centers for Medicare and Medicaid Services were used to identify claims associated with PT interventions. Our analyses describe trends in PT claims, CPT codes, cancer certifications by subtype, and number of members with Category A cancers and PT claims. Results/Anticipated Results: Since the Program’s inception in 2011, PT claims had gradually increased except for in 2020 when there was a sudden decrease, most likely due to the interruption of in-person services due to the COVID-19 pandemic. From 2021 to 2023, PT claims began to increase again. The most common types of PT interventions were therapeutic exercises, manual therapy, and neuromuscular reeducation. In 2023, the most recent year of full data available, Category A cancers made up 38% of all cancer certifications, with prostate and breast cancer being the most common. Category A cancers were evaluated together due to prior existing evidence outlining significant benefit from PT intervention. In total, Category A cancers represent over 14,000 Program members. Less than 1% of members with a Category A cancer had a PT related claim in 2023. Discussion/Significance of Impact: The mission of the Program is to provide quality and compassionate medical care and treatment to our members. Better understanding the utilization of PT services provided by the Program will allow us to increase awareness and support of interventions for members of our Program who could benefit from PT services.

Objectives/Goals: Alzheimer’s disease (AD) has limited treatments and an extremely high rate of clinical trial failure. Through a collaborative effort, Agomelatine (AGO) was identified as having repurposing potential for AD. This study sets out to evaluate the preclinical potential of AGO for the treatment of AD. Methods/Study Population: The TgF344-AD rat model (expresses human mutant “Swedish” amyloid-precursor protein and a Δ exon 9 presenilin 1) was used to test AGO’s potential to reduce cognitive deficits and neuropathology. The model was chosen due to its age-dependent progressive AD pathology and cognitive decline. Treatment with AGO at ~10 mg/kg body weight/day began at 5 months of age (pre-pathology) and continued until 11 months of age when cognitive testing (active place avoidance task) and tissue collection occurred. Immunohistochemistry was used to evaluate amyloid beta plaque burden and microglial response in the hippocampus. Results/Anticipated Results: AGO-treated female TgF344-AD rats showed reduced cognitive deficits with an increased latency to first entrance in aPAT testing compared to nontreated transgenic littermates. There were no differences between the cognitive performance of AGO treated and untreated male TgF344-AD rats. Interestingly, this reduced cognitive deficit did not correlate with decreased amyloid beta pathology in female AGO-treated rats yet male transgenic treated rats did have decreased amyloid burden in the dentate gyrus (DG) of the hippocampus. AGO modulated microglial activation in the DG of female transgenic rats. Discussion/Significance of Impact: AGO reduced cognitive deficits in females, but did not change their amyloid burden. This suggests that AGO could increase resilience to amyloid deposition in female rats. With the recent development of amyloid targeting drugs, novel non-amyloidogenic treatments have a large translational potential.