Objectives/Goals: To identify the genomic mechanisms underlying cross-species regulation of longevity among mammals and birds and to characterize the impact of those conserved pathways on human aging. More broadly, this study aims to develop a novel evolutionary approach to understand the genetics of complex traits. Methods/Study Population: High-quality genome sequences for 194 bird species and 295 mammal species with reliable longevity and body size data were obtained from publicly available resources. The data include coding sequence alignments of 16,863 mammalian and 14,565 avian one-to-one orthologous genes. Gene-wise relative evolutionary rates (RERConverge) and maximum likelihood phylogenetics (PAML) were computed to assess for evidence of purifying selection and positive selection for longevity. As part of ongoing analysis, human orthologs of selected results will be examined in the UK Biobank for validation. Results/Anticipated Results: Preliminarily, we have found a signal of concordant positive selection between two classes of vertebrates separated by hundreds of millions of years. Several genes show signals of positive selection in long-lived species of both birds and mammals. Ongoing work focuses on elucidating the relationship between relative evolutionary rates and positive selection, the overlap in selection signature between long-lived animals with large body sizes and those who are exceptionally long lived for their body size, and further elaborating on convergence between mammals and birds. Discussion/Significance of Impact: Currently, most scientific knowledge about aging is from experiments on short-lived model organisms. By systematically studying the genomes of long-lived mammals and birds, we aim to develop a new method for studying complex traits and uncover novel insights into the mechanisms of longevity.

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.

We conducted a pilot study of implementing community health workers (CHWs) to assist patients with hypertension and social needs. As part of clinical care, patients identified as having an unmet need were referred to a CHW. We evaluated changes in blood pressure and needs among 35 patients and conducted interviews to understand participants’ experiences. Participants had a mean age of 54.1 years and 29 were Black. Twenty-six completed follow-up. Blood pressure and social needs improved from baseline to 6 months. Participants reported being accepting of CHWs, but also challenges with establishing a relationship with a CHW and being unclear about their role.

Community advisory boards (CABs) have traditionally been formed in the context of discrete projects and served to support community protections within the confines of the associated investigation(s). However, as funding bodies increasingly prioritize health equity, CABs have shifted – evolving into long-running organizations with broader scope and value. An emerging cornerstone of these project-independent boards (PICABs) has been the formation of “Research Review Boards” (RRBs). While unified in their goal of promoting community protection and representation in health research, it is unknown to what degree RRBs differ on key features including membership, leadership, service reach, and – crucially – impact. A scoping review was conducted according to PRISMA-ScR guidelines to analyze current practices for RRBs. Of screened articles (n= 1878), 25 were included, corresponding to 24 unique RRBs. Findings indicated overlaps in the stated missions, funding structures, and processes of most RRBs. Differences in membership composition, location, service-reach, leadership structures, evaluation procedures, and perceived impact were evident. Where data is available, RRBs receive positive endorsement from both internal members and external users. Standardization of evaluation procedures is needed to fully quantify impact. Additional challenges to sustainability, communication, and conflicts (e.g., of interest, commitment, and power differentials) merit further consideration.

Objectives/Goals: Infectious keratitis is the leading cause of corneal blindness worldwide, causing two million cases of monocular blindness per year. Of these cases, developing countries are disproportionately affected, in part due to sociodemographic disparities. Our study examined risk factors for severe keratitis presentation in a South Indian population. Methods/Study Population: 156 patients aged ≥ 16 years with clinically diagnosed infectious keratitis presenting to Aravind Eye Hospital in Pondicherry, India, from January 1, 2023 to July 31, 2024, were retrospectively reviewed. Univariate logistic regression was used to evaluate associations between specific potential risk factors (including age, sex, awareness of keratitis, travel distance to hospital, education level, missed work wages, and ability to afford care) and keratitis clinical severity (defined using thresholds of poor visual acuity, size of stromal infiltrate measured on slit lamp examination, occurrence of perforation, and need for corneal transplant surgery). Individual risk factors found to be significant were incorporated into a multivariable logistic regression model. Results/Anticipated Results: We anticipate that the results of this study will identify multiple risk factors for more severe keratitis presentations among patients at baseline. We expect these factors to include increased travel distance from the patient’s home to the base hospital, delays between time of diagnosis and initiation of treatment, treatment nonadherence, lower educational levels, lack of familiarity with keratitis, treatment and transportation costs, increased time off work, and missed work wages, among others. Discussion/Significance of Impact: This study elucidates barriers to early keratitis diagnosis in a low-resource setting. Study findings can inform strategies to improve keratitis prevention using decentralized care approaches such as community eye screenings and expanding outreach via vision centers. Such strategies can improve timely access to care for vulnerable populations.

Objectives/Goals: Immune checkpoint inhibitors (IO) have dramatically improved survival outcomes in patients with metastatic melanoma. Still, many patients do not respond these treatments, and others may experience harmful adverse events (irAEs). Thus, there an unmet need for biomarkers for real-time monitoring and management of patients exposed to IO therapies. Methods/Study Population: Serial serum samples were collected from patients with BRAFV600-mutant metastatic melanoma treated with ipilimumab/nivolumab (IO, n = 14) or dabrafenib/trametinib (TT, n = 10). Methylated cell-free DNA (cfDNA) was isolated and sequenced using enzymatic methyl-seq. We develop a robust computational pipeline to identify the top 250 cell-type specific regions of differential methylation (DMRs) across 24 cell-types. Using these differentially methylated regions, a deconvolution tool was developed to determine the abundance of cell type-specific cfDNA in patient serum, and changes in abundance were tracked over treatment time-course to assess response treatment and identify signals of adverse events. Results/Anticipated Results: We demonstrated improved precision in DMR detection evidenced by a higher area under the receiver operator characteristic curve (AUROC) of 0.85 on average. Pathway and functional annotation analysis revealed melanocyte-specific methylation marker regions regulated genes related to melanocyte development and differentiation, including MITF, SOX9/10, and FOXD3. We show these regions are conserved through the transformation to malignant melanoma, indicating melanocyte cfDNA abundance can be used as a marker for tumor burden. We characterize the dynamics of melanocyte-derived cfDNA over the course of treatment in responders and nonresponders to both IO and TT. We observe that changes in concentrations of cfDNA from other cell types correlate with clinically observed irAE-mediated damage to normal tissue. Discussion/Significance of Impact: We demonstrated the utility of decoding the origins of cfDNA fragments obtained from serial liquid biopsy samples. Using cell-specific methylation marks, we identified a signature from the primary melanoma to assess response to treatment, while also obtaining a signal from other tissues throughout the body to monitor immune related adverse events.

Objectives/Goals: We investigate how the gut microbiome protects against arsenic toxicity, showing antibiotic perturbation increases toxicity and causes interindividual susceptibility to a sepsis-like disease state in mice. Here, we aim to understand how baseline microbiomes from various mouse vendors impact these outcomes and characterize the observed disease. Methods/Study Population: We developed a novel mouse model where mice are exposed to an antibiotic (cefoperazone) for 2 days, followed by co-exposure to the antibiotic and 100 ppm arsenate. So far, we have evaluated C57BL/6N mice from MSU’s in-house colony, Taconic Biosciences (TAC), and Jackson Labs (JAX), along with C57BL/6J mice from JAX. To determine if the baseline microbiome drives inter-vivarium differences, we established in-house breeding colonies of TAC- and JAX-origin mice at MSU. This allowed us to assess whether, when housed under identical conditions, these mice still show differences in mortality based on their original microbiomes. To characterize the arsenic-induced sepsis-like disease, we performed blood biochemistry assays to quantify the white blood cell populations, and sepsis biomarkers used in clinical settings. Results/Anticipated Results: We observed differences in survival rates between genetically identical mice from MSU (45%), TAC (30%), and JAX (2.5%) in our model. From this, we characterized the baseline composition of the gut microbiomes of these mice and found they were significantly different from each other. We are still awaiting results from our in-house TAC and JAX experiments but expect them to have similar gut microbiome compositions to those directly purchased from TAC and JAX and respond similarly. In our blood biochemistry analysis, we found sick mice presented with low WBC counts and notable biomarkers indicative of liver, heart, and kidney distress. We also anticipate that 16S sequencing results of cecal contents will further support findings by providing evidence of a bacterial infection in the ceca of sick mice. Discussion/Significance of Impact: Collectively, our work demonstrates that antibiotic perturbation of the gut microbiome induces an inter-individual and inter-vivarium susceptibility to an arsenic-induced sepsis-like disease state. This work highlights the importance of considering antibiotic use in the risk assessment of arsenic to better protect the health of those exposed.

Objectives/Goals: The overall goal of this project is to determine bacterial transcriptional signatures from clinical sputum and assess their potential to monitor treatment response and predict the outcome of drug therapy in patients with tuberculosis (TB). Methods/Study Population: We are developing a novel transcript capture sequencing (TC-Seq) approach to sequence the mRNA of Mycobacterium tuberculosis (Mtb) and analyze transcriptomes from clinical samples containing minimal amounts of bacterial RNA. This protocol generates single-stranded biotinylated probes from Mtb DNA. Probes are hybridized to and allow enrichment of Mtb-specific mRNA within next-generation RNA sequencing libraries. We will apply TC-Seq to sputum samples collected throughout an 18-month Phase II clinical trial investigating response to TB treatment to compare the transcriptome of Mtb between patients whose treatment results in cure or relapse. Results/Anticipated Results: We have refined a technique to generate biotinylated probes starting from DNA of lab grown Mtb. This protocol achieves robust and unbiased sampling of the Mtb transcriptome from mixed samples containing both human and Mtb RNA. Preliminary sequencing of clinical sputum collected pretreatment has generated 1–4 million Mtb-specific reads, a sequencing depth that allows examination of the entire bacterial transcriptome. We will measure differential gene expression before and during treatment as well as between cure and relapse cases. These results will allow us to characterize bacterial response to treatment and identify bacterial markers that correlate with relapse. Discussion/Significance of Impact: Understanding Mtb activity during treatment will offer new ways to assess the efficacy of different treatment regimens. Crucially, identifying clear bacterial markers that demarcate a cure or relapse outcome will have a significant impact on determining patient eligibility for shorter drug therapy.

The National Institutes of Health (NIH) diversity supplements represent an opportunity to enhance diversity in the biomedical research workforce. Despite their potential impact, practical barriers prevent effective use of these resources. The Altman Clinical and Translational Research Institute (ACTRI) at the University of California San Diego (UCSD) developed and implemented of an institutional repository and support system to improve diversity supplement applications. The centralized repository and support system incorporated three strategies: (1) a secure web-based repository housing successful examples of supplements, (2) match making for diversity supplements and mentors, and (3) web-based resources for potential applicants. The repository was implemented in June 2021 and includes 51 supplement samples across 14 divisions of NIH as of November 2024. The repository has been accessed by 51 potential applicants since implementation in 2021. Few investigators have requested match making. Early indicators show a doubling in diversity supplement applications compared to pre-implementation period at UCSD. We outlined our approach to addressing some of the barriers in diversity supplement applications, which could provide a model for other institutions. Pending solutions to some of the lessons learned, coordinated efforts aimed at diversity supplements could be a practical approach toward a more diverse biomedical research workforce.

Objectives/Goals: This poster discusses key methodological and theoretical issues in translation and implementation for improving HPV vaccine recommendations in clinics serving rural communities. Methods/Study Population: Leveraging implementation science, the study of how to improve the uptake of evidence-based practices, this pilot study uses a mixed-methods effectiveness-implementation design to engage local experts in identifying a bundle of locally-tailored implementation strategies to facilitate uptake of evidence-based HPV vaccine recommendations. In partnership with the University of Arkansas for Medical Sciences Rural Research Network, we will follow an evidence-based quality improvement process to develop locally tailored implementation strategies, which we will then evaluate for acceptability, feasibility, and effectiveness. Results/Anticipated Results: This study aims to generate actionable insights into the design and implementation of tailored, evidence-based communication strategies that can be scaled to improve HPV vaccine uptake in rural communities. Findings from this pilot study will be used to support a future full-scale Hybrid-Type 3 effectiveness-implementation trial to evaluate the bundle of tailored implementation strategies. Discussion/Significance of Impact: By addressing the rural-specific determinants of evidence-based HPV vaccine recommendations, this research will contribute to a deeper understanding of how to support high-quality, evidence-based provider recommendations in rural, underserved communities, and will mitigate rural disparities in HPV-related cancers.

Objectives/Goals: The timing of neurosurgery is highly variable for post-hemorrhagic hydrocephalus (PHH) of prematurity. We sought to utilize microvascular imaging (MVI) in ultrasound (US) to identify biomarkers to discern the opportune time for intervention and to analyze the cerebrospinal fluid (CSF) characteristics as they pertain to neurosurgical outcome. Methods/Study Population: The inclusion criteria for the study are admission to the neonatal intensive care unit (NICU) with a diagnosis of Papile grade III or IV. Exclusion criteria are congenital hydrocephalus and hydrocephalus secondary to myelomeningocele/brain tumor/vascular malformation. We are a level IV tertiary referral center. Our current clinical care pathway utilizes brain US at admission and at weekly intervals. Patients who meet certain clinical and radiographic parameters undergo temporary or permanent CSF diversion. Results/Anticipated Results: NEL was implemented at our institution for PHH of prematurity in fall 2022. To date, we have had 20 patients who were diagnosed with grade III or IV IVH, of which 12 qualified for NEL. Our preliminary safety and feasibility results as well as the innovative bedside technique pioneered at our institution are currently in revision stages for publication. Preliminary results of the MVI data have yielded that hyperemia may confer venous congestion in the germinal matrix, which should then alert the neurosurgeon to delay any intervention to avoid progression of intraventricular blood. With regard to CSF characteristics, we anticipate that protein, cell count, hemoglobin, iron, and ferritin will decrease with NEL. Discussion/Significance of Impact: The timing of PHH of prematurity is highly variable. We expect that MVI will offer radiographic biomarkers to guide optimal timing of neurosurgical intervention. A better understanding of CSF characteristics could potentially educate the neurosurgeon with regard to optimal timing of permanent CSF diversion based on specific CSF parameters.

The need for collaborative and transparent sharing of COVID-19 clinical trial and large-scale observational study data to accelerate scientific discovery and inform clinical practice is critical. Responsible data-sharing requires addressing challenges associated with data privacy and confidentiality, data linkage, data quality, variable harmonization, data formats, and comprehensive metadata documentation to produce a high-quality, contextually rich, findable, accessible, interoperable, and reusable (FAIR) dataset. This communication explores the experiences and lessons learned from sharing National Heart Lung and Blood Institute (NHLBI) COVID-19 clinical trial (including adaptive platform trials) and cohort study datasets through the NHLBI BioData Catalyst® (BDC) ecosystem, focusing on the challenges and successes of harmonizing these datasets for broader research use. Our findings highlight the importance of establishing standardized data formats, adopting common data elements and creating and maintaining robust data governance structures that address common challenges (i.e., data privacy and data-sharing limitations resulting from informed consent). These efforts resulted in a set of comprehensive and interoperable datasets from 5 clinical trials and 13 cohort studies that will enable downstream reuse in analyses and collaborations. The principles and strategies outlined, derived through experience with consortia data, can lay the groundwork for advancing collaborative and efficient data sharing.

The COVID-19 pandemic laid bare the inequities in U.S. healthcare in ways that captured public attention and reinforced the need to view all of healthcare through an equity lens. It also exposed global inequities in access to healthcare technologies. At Rockefeller University, we participate in the entire spectrum of translational research, but our focus is in the areas of basic research and new methods to prevent, diagnose, and treat disease, extending to proof of concept preclinical and Phase 1 studies. Since we believe that all phases of translational research should have an equity lens, we have instituted an initiative to encourage thought and planning about global equitable access to discoveries made by our trainee Clinical Scholars and faculty, even at the earliest phases of basic research. Assuring global equitable access to new technologies requires addressing at least 3 different aspects of new technology: 1. Patenting and licensing, 2. Manufacturing, and 3. Dissemination and implementation in low- and middle-income countries. In this review, I focus on patenting and licensing and offer ten questions for inventors to consider in discussing licensing their technologies with technology transfer officers to maximize equitable global access to the technologies they create.

Objectives/Goals: To determine the role of carcinoembryonic antigen cell adhesion molecule 6 (CEACAM6) in signaling cascade and interaction with other cell surface proteins in human epithelial solid tumors such pancreatic adenocarcinoma and colon cancer Methods/Study Population: In this study, we employed three-dimensional (3D) tumor models to replicate the in vivo tumor microenvironment better, allowing for a more accurate assessment of cellular responses compared to traditional two-dimensional (2D) cultures. We used immunoprecipitate to pull down the CEACAM6 protein and investigate the integrins expression level. Results/Anticipated Results: The expression and functional activity of CEACAM6 are susceptible to modulation by various surface proteins, leading to notable alterations in cellular behavior. Integrins, particularly Integrin B4, are one such protein whose expression is influenced by CEACAM6-mediated intracellular signaling cascades, suggesting a complex interplay that enhances CEACAM6 activation.The 3D models facilitate cell–cell interactions, enabling tumor cells to proliferate and undergo metabolic changes that reflect actual tumor biology. Thereby enhancing the relevance of crosstalk between CEACAM6 and integrins. These findings underscore the potential of CEACAM6 as a promising therapeutic target. They reveal a molecular mechanism that could inform the development of innovative therapeutic strategies in cancer. Discussion/Significance of Impact: These findings underscore the potential of CEACAM6 as a promising therapeutic target, revealing a novel molecular mechanism that could inform the development of innovative therapeutic strategies for pancreatic and colon cancer and potentially other malignancies.

Objectives/Goals: Understanding systematic review results help prioritise more valuable studies. However, evaluating whether a systematic review has conclusively answered a question is difficult, and it is unclear which tools are available for such assessments. Thus, we mapped the extent of methods for determining the conclusiveness of systematic review results. Methods/Study Population: We searched Medline (Ovid), EMBASE (Ovid), and Web of Science to find papers with methods to determine whether systematic review results were conclusive or should be updated. The characteristics of primary references for included methods are presented. We classified and summarized available methods. Results/Anticipated Results: A total of 58 unique methods were identified. Many have been published since 2010 and often did not include a worked example. We found 25 mathematical methods for the conclusiveness of meta-analyses, which included cumulative meta-analysis, fail-safe number, fragility index, prediction and machine learning model, simulation-based power, conditional power, and graphical approaches. There were 15 methods for the conclusiveness of cumulative meta-analyses, such as quality control approach, trial sequential analysis, sequential meta-analysis, and law of iterated logarithm. And, 18 methods assessed the conclusiveness of systematic reviews: GRADE framework, the strength of a body of evidence approach, methods for assessing the need to update a systematic review, and methods for specific clinical domains. Discussion/Significance of Impact: We found a wide range of methods that can be used to determine the conclusiveness of systematic review results. End-users of systematic reviews can review our results to find the most appropriate methods for their contexts and decisions.