Composite endpoints can encode multiple pieces of information and are increasingly adopted in clinical trials. Advocacy for using composite endpoints began decades ago in cardiovascular trials, leading to incorporation of patient-oriented outcomes and consideration of a hierarchical ranking system. The use of composite endpoints in coronavirus disease (COVID-19) trials has evolved similarly. We conducted a literature review to investigate the use of composite endpoints in acute heart failure and COVID-19 clinical trials. The results showed more frequent use of patient-oriented outcomes and ordinal composite endpoints in COVID-19 trials, which might be driven by global consensus on a set of common outcome measures.

ABSTRACT IMPACT: I hope that our work will improve surveillance endoscopy experiences, by engaging both patients and providers. OBJECTIVES/GOALS: A large proportion of colonoscopies are performed for post-polypectomy surveillance. Data show that there is overuse of surveillance for low-risk adenomas (LRAs), which can be attributed to patient and provider factors. The objective is to understand patient and provider perspectives for decision-making for LRA surveillance colonoscopy. METHODS/STUDY POPULATION: Semi-structured, one-on-one virtual interviews of patients and providers are currently being conducted at Richard L. Roudebush VA Medical Center and Eskenazi Health. Using a criterion sampling approach, we identified patients 50-75 years of age who had a screening colonoscopy with finding of LRAs, and providers in primary care and gastroenterology, at each site. We plan to recruit at least 8 patients and 8 providers from each site until thematic saturation. Domains that will be covered include: perceived involvement with surveillance decision-making; experiences with, and preferences for, communication about test results; and barriers and facilitators to undergoing colonoscopy. A 3-phase approach, comprising immersion, reduction, and interpretation, is being used to collect and analyze data. RESULTS/ANTICIPATED RESULTS: This study is currently in the recruitment phase and results will be forthcoming. DISCUSSION/SIGNIFICANCE OF FINDINGS: Understanding decision-making for LRA surveillance colonoscopy will inform future interventions to improve endoscopic resource use and efficiency while improving patient and provider experiences with endoscopic care coordination.

Objectives/Goals: The STEM-Coaching and Resources for Entrepreneurial Women (CREW) Program was developed to promote systemic change in entrepreneurship by engaging women faculty and post-doctoral fellows through educational opportunities, mentorship, and professional coaching to close the gender gap in entrepreneurial leadership. Methods/Study Population: We solicited applications from women at the junior faculty or post-doctoral fellowship stage. Applications were reviewed by the CREW leadership and mentors. Applicants accepted into the program participated in monthly large group coaching sessions, one-one-one coaching, and small group mentoring sessions. Applicants also completed an online entrepreneurship course and participated in a shark tank-like competition during annual innovation awareness month. Coaches were professionally credentialed and oriented to the biomedical context. Entrepreneurial experts served as mentors. Our program evaluation used the RE-AIM framework. The evaluation included assessments of the program’s ability to reach a diverse population, satisfaction with program offerings, and changes in entrepreneurship outcomes. Results/Anticipated Results: To date, three cohorts consisting of 35 women from 13 states participated in the program. Results from Cohort 1 show significant improvements in self-assessed confidence in entrepreneurial activities and high satisfaction with the program. All measures of confidence showed increases, with the largest gains in the areas of “estimating customer demand for new products or services”; “Clearly and concisely explaining my business idea in everyday terms”; and “Getting others to identify with and believe in my vision and plans for a new business”. The percentage of participants who have invention disclosures significantly increased from 33.3% to 83.3% after completing the program. The percentage who launched start-up companies also increased from 17% to 33%. Cohort 2 results will be available before the conference. Discussion/Significance of Impact: Participants in the CREW program showed improvement in all measures of evaluation. The program is unique in complementing mentoring with coaching. Because the coaching component takes a holistic approach by encouraging women to craft a vision for their work and life, it provides value that will last beyond entrepreneurship activities.

OBJECTIVES/SPECIFIC AIMS: We previously developed a translationally relevant model of temporal lobe epilepsy (TLE) in which glutamine synthetase is irreversibility inhibited by methionine sulfoximine (MSO), resulting in spontaneous seizures and dentate hilar neuron loss. The objective of this study was to determine the effects of chronic BCAA ingestion on neuronal viability in the dentate hilus in the MSO model of TLE. METHODS/STUDY POPULATION: Sixteen rats were randomly divided into 2 groups: 8 rats drank a 4% aqueous solution of all 3 BCAAs (BCAA group) ad libitum for 31 days, and the other 8 rats drank regular water (control group) for the same period. After 10 days of drinking, a microinfusion cannula (Alzet osmotic pump, model 2004) was surgically implanted in the right dentate gyrus to continuously infuse MSO at a rate of 0.625 g/hour for 28 days. After 31 days of drinking, rats were perfused transcardially with 0.9% NaCl followed by 4% paraformaldehyde in phosphate buffer. The brains were removed and fixed, sectioned on a Vibratome at 50-μm thickness, and were mounted on a gelatin-coated slides and stained with NeuN. Neuron counts in the hilar region were performed ipsilateral and contralateral to the infusion site using a stereological technique. RESULTS/ANTICIPATED RESULTS: Rats in the BCAA group had 37% fewer neurons in the ipsilateral dentate hilus than the control group (5.8×10−4±6.8×10−5 vs. 8.9×10−4±5.6×10−5 cells, respectively, p<0.01). Similarly, rats in the BCAA group had 39% fewer neurons in the contralateral dentate hilus than the control group (5.0×10−4±5.8×10−5 vs. 7.0×10−4±3.4×10−5 cells, respectively, p=0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrates that chronic ingestion of BCAAs aggravates hilar neuronal loss in a translationally relevant rodent model of MTLE. This study gives important insight into how BCAAs may affect neuronal viability. Although the role of BCAAs on seizure activity is poorly understood, these results suggest that BCAAs may play an important role in neurochemical modulation and neurotoxicity.

OBJECTIVES/GOALS: Project Extension for Community Health Outcomes (ECHO), a telehealth model, was launched at Penn State University in 2018 to connect specialists with community providers to provide education on best clinical practices. We aim to describe clinical topics covered and relevant provider level outcomes. METHODS/STUDY POPULATION: The heart of the ECHO model is a hub-and-spoke knowledge-sharing system. The ECHO model has four core principles: 1) use technology to leverage scarce resources; 2) share best practices to reduce disparities; 3) employ case-based learning to master complexity; 4) monitor outcomes to ensure benefit. Unlike telemedicine, where outside specialists assume the care of the patient, Project ECHO is a guided learning community aimed at practice improvement: providers receive mentoring and feedback on de-identified patient cases, strengthen their skillset, and retain responsibility for their patients. RESULTS/ANTICIPATED RESULTS: Clinical topics launched include Medication Assisted Treatment for Opioid Use Disorder, Ehlers Danlos Syndrome, Polyneuropathy, and Dementia. In addition, we launched a nutrition-focused ECHO with Boy Scout summer camp leaders in 26 states, reaching 107,347 scouts. Over the past year we have reached 118 clinicians in 62 clinics within 19 counties in Pennsylvania, providing a total of 268 CME hours. These providers have treated 2,294 patients and reported increased knowledge (94%), decreased sense of professional isolation (86%), and improvement in ability to provide patient care (92%) following completion of an ECHO series. DISCUSSION/SIGNIFICANCE OF IMPACT: Project ECHO is a powerful telehealth model providing mentorship and education to clinicians, encouraging them to treat more complex cases in their primary care clinics. As a result, patients receive higher quality care when they need it, and close to home, particularly important in rural areas.

OBJECTIVES/GOALS: Adolescent and young adult (AYA) patients, diagnosed with cancer between 15 and 39 years of age, often have worse outcomes compared to younger children and older adults. We will characterize age differences in acuity and severity of illness at initial presentation, as a measure for health outcomes, in patients with new diagnoses of leukemia or lymphoma. METHODS/STUDY POPULATION: We will perform a single institution-based, retrospective analysis of a cohort of pediatric oncology patients at Children’s Healthcare of Atlanta (CHOA) who were diagnosed with leukemia or lymphoma from 2010 to 2018. Data will be abstracted from the CHOA medical record and cancer registry. We will construct a severity score, with one point for 1) admission to the Intensive Care Unit (ICU), 2) ICU-level requirements at admission, 3) stage IV or metastatic disease, and 4) white blood cell count over 100,000/microliter. We will associate this score with age at presentation using logistic regression models among patients overall and stratified by disease type. All regression models will adjust for confounders (e.g., health insurance, race and ethnicity, and local-area social vulnerability level). RESULTS/ANTICIPATED RESULTS: Previous literature has shown increased cancer incidence and mortality in AYAs compared to younger children. Therefore, we hypothesize that AYA patients with a new diagnosis of leukemia or lymphoma will have a higher acuity and severity of illness at presentation than younger patients. We conducted a pilot study analyzing acuity and severity of illness by age, race, ethnicity, and insurance status. Bivariate comparisons suggested patients aged 10 and older were as likely as those younger to have a severity score of more than 0 at diagnosis (OR=0.661 , 95% CI 0.43-1.01). However, this result was collapsed across all hematologic malignancies, did not encompass the entire cohort, and did not address possible confounders; we anticipate our estimates will be different taking these factors into account. DISCUSSION/SIGNIFICANCE: Our work will be the first steps in creating a validated tool to understand disease acuity and severity. By using this tool to characterize presentation by disease type and age, we will identify unmet needs prior to an initial diagnosis of cancer. Our findings inform strategies toward narrowing age disparities in outcomes for AYA hematologic cancers.

OBJECTIVES/SPECIFIC AIMS: Platelets govern signal-dependent inflammatory responses by leukocytes. Although dysregulated inflammation is common in older adults, platelet-leukocyte signaling events and downstream inflammatory gene synthesis in aging is not known. METHODS/STUDY POPULATION: Highly-purified platelets and monocytes were isolated from healthy older (age>60, n=27) and younger (age<45, n=36) adults and incubated together in autologous and nonautologous conditions. Inflammatory gene synthesis by monocytes, basally and in the presence of activated platelets, was examined. Next-generation RNA-sequencing allowed for unbiased profiling of the platelet transcriptome in older and younger adults. Differentially expressed candidates in aged platelets were validated and recombinant granzyme A (in the presence and absence of TLR4 and Caspase-1 inhibition) identified putative ligands controlling inflammatory gene synthesis. RESULTS/ANTICIPATED RESULTS: In unstimulated or activated conditions, monocyte chemoattractant protein 1 (MCP-1) and interleukin-8 (IL-8) synthesis by monocytes alone did not differ between older and younger adults. However, in the presence of autologous activated platelets, monocytes from older adults synthesized significantly greater MCP-1 (867.150 vs. 216.36 ng/mL, p<0.0001) and IL-8 (41.5 vs. 9.2 ng/mL, p<0.0001) than younger adults. Nonautologous, or switch experiments, demonstrated that aged platelets were sufficient for upregulating MCP-1 and IL-8 synthesis by monocytes. Surprisingly, classic platelet proteins known to signal to monocytes and induce MCP-1 synthesis (p-selectin, RANTES, and PF4) were not increased in platelets from older adults. Using RNA-seq followed by validation via RT-PCR and immunoblot, we identified candidate platelet molecules increased in aging that mediate platelet-monocyte signaling and pro-inflammatory gene synthesis. We confirmed that granzyme A (GrmA), a serine protease not previously identified in platelets, is present in human platelets at the mRNA and protein level. GrmA is secreted by activated platelets in signal-dependent fashion. Moreover, GrmA in platelets is significantly increased in aging (~9-fold vs. younger adults). Blocking GrmA inhibited MCP-1 and IL-8 synthesis in older adults. Finally, we uncovered that platelet GrmA signaling to monocytes is regulated through TLR4 and Caspase-1. DISCUSSION/SIGNIFICANCE OF IMPACT: Human aging is associated with reprogramming of the platelet transcriptome. A previously unrecognized protein in platelets, GrmA, is increased in aging and causes increased MCP-1 and IL-8 gene synthesis by target monocytes in a TLR4 and Caspase-1 dependent mechanism. Increased platelet GrmA in aging may contribute to injurious inflammatory responses common in older adults.

ABSTRACT IMPACT: Framework is designed to aid selection of implementation strategies to promote adoption and sustainability of EBP to improve health care quality, safety and value. OBJECTIVES/GOALS: An application-oriented implementation framework based on Diffusion of Innovation theory, identified 81 strategies for clinician-use within four implementation phases. The goal of this research was to further specify strategies based on emerging implementation science and establish external validity. METHODS/STUDY POPULATION: An iterative mixed-methods process guided framework revisions. First, individuals (n=1,578) requesting use of the framework over the last seven years were sent an electronic questionnaire. Evaluation captured usability, generalizability, accuracy of phases, and implementation phases for each of 81 strategies. Second, nurses who use the framework pile sorted strategies for multidimensional scaling and hierarchical analysis using Anthropac software. Third, a panel of five EBP/implementation experts used data and a consensus process to add clarity with the naming, and further specify strategies. RESULTS/ANTICIPATED RESULTS: Survey respondents (n = 127, 8% response) were nurses (94%), at least Master’s educated (94%), from health systems (52%) or academia (31%), in the U.S. (84%). The framework, rated on a four-point scale (1 = not/strongly disagree to 4 = very/strongly agree; reported are ratings 3 and 4) was deemed useful (92%), generalizable (100%), and with accurate timing (96%). 51 participants linked strategy timing to a single phase (54 strategies, 66.7%, p<0.05, Cochran’s Q); most strategies (30) matched the original model. Pile sorting (n=23) generated a concept map and hierarchical clusters of groups. Experts used these data and implementation science to specify each strategy and revise the framework. DISCUSSION/SIGNIFICANCE OF FINDINGS: The Iowa Implementation for Sustainability Framework (IISF) offers a typology to guide implementation for healthcare improvements. This study specifies 77 implementation strategies, confirms four phases, identified 10 domains, and begins to establish external validity for the framework.

Objectives/Goals: The goal of the Accelerated Staff Assist Program (ASAP) is to help fill staffing gaps in clinical and translational research. The program offers centralized staff to fill short-term needs including project startup prior to hiring, project implementation, parental leave, and Spanish-language research support. Methods/Study Population: Promotion of ASAP in the first year was targeted to Penn State College of Medicine faculty to determine demand. Current outreach includes promotion to all Penn State campuses through PSCTSI’s newsletter, seminars, and presentations. Consultation intake data includes a project summary, staffing needs, and funding information. A project agreement, charter, and budget are drafted and agreed to by the principal investigator before work begins. Assessments occur at 6 and 12 months to assess satisfaction of the consultation process and quality of staff support, Net Promoter Score, and qualitative feedback about the program. Results/Anticipated Results: In its first year, ASAP conducted 26 consultations and supported 10 projects. Approximately 75% of staff effort has been recovered through agreements during the most recent 6 months. In anticipation of project delays, we found that booking FTE up to 115% results in a consistent recovery of 75% FTE. Consultee needs include short-term, partial effort support (76% of requests), project start-up (12%), staff leave (4%), and other (8%). ASAP staff tasks include participant recruitment and retention, data collection and analysis in English and Spanish, and community engagement for recruitment. Program assessments with research teams are ongoing; preliminary qualitative and quantitative data are unanimously positive. Discussion/Significance of Impact: The ASAP program has demonstrated fiscal success and sustainability in its first year. Hiring of a well-qualified, bilingual staff person allowed for quick onboarding and project support within one month of hire. ASAP enables CTSI to fulfill our mission by providing critical support for research teams to meet objectives and achieve equity goals.

OBJECTIVES/SPECIFIC AIMS: Calls to break the silence around the effects of racism on health are growing. Few researchers have examined the relationship between medical student characteristics and students’ comfort, motivation, and skill to discuss racism. This paper examines medical student characteristics associated with readiness to talk about racism among first-year medical students at the University of Minnesota. METHODS/STUDY POPULATION: In February 2017 prior to a lecture on racism and health, we invited first year medical students to participate in a web-based survey about their experiences and comfort discussing racism. We calculated descriptive statistics and measured differences by student race (White vs. Asian vs. Black/multiracial/other) and undergraduate major type (STEM vs. non-STEM) using χ2 tests for variables with categorical responses and generalized linear regression models with pairwise comparisons (i.e., 2-sample t-tests) for variables with continuous responses. RESULTS/ANTICIPATED RESULTS: (n=107/163). The majority of students were male (53%); White (75%); and majored in STEM majors in college (85%). College major was not associated with race. Students’ responses to multiple items suggest that the vast majority perceived racial inequality as a major problem in the United States. Race was significantly associated with only 1 of these items. Specifically, 100% (16/16) of Black/multiracial/other students [under-represented minority (URM) students] reported “too little attention” is paid to race and racial issues, while only 53% of White students (42/79) and 55% of Asian students (6/11) chose this response. Students with non-STEM majors and students who identified as URM students reported talking about racism with friends more often than STEM majors and white students, respectively. In conversations about race at school, two-thirds of students were concerned that they might unintentionally offend others or be misunderstood. However, non-STEM majors and URM students were significantly less worried that they would unintentionally offend others in conversations about race at school than STEM majors and white students. Larger percentages of URM students (50%) than White students (25%) were afraid that others would not respect their views because of their race. White students were more afraid that they might that they would be called racist than URM students. DISCUSSION/SIGNIFICANCE OF IMPACT: Many students find it challenging to discuss race and racism in medical education settings. URM students and non-STEM majors reported greater frequency talking about racism with friends and appear to be less anxious in conversations about racism than White students and STEM majors respectively. Given non-STEM majors' greater psychological safety discussing racism, future research should explore whether non-STEM majors are better prepared and more motivated to address racial disparities in health and health care than STEM majors. Such research could have important implications for medical school admissions.

OBJECTIVES/GOALS: We conducted a review of CTSA websites to understand the current landscape for CRP institutional professional development and training revealed in the CTSA hub websites. METHODS/STUDY POPULATION: We accessed and reviewed 59 currently funded CTSA hub websites for evidence of CRP training opportunities. Parameters reviewed included: 1) opportunities were specified for CRPs versus K and T trainees; 2) mandated training; 3) leveling; 4) delivery methods/resources; 5) public accessibility; 6) unique features. The website reviews informed a REDCap survey sent to the CTSA Administrators (n = 149) and the Coordinator Taskforce (n = 105) listservs to gain additional knowledge of CRP training available at the institution. A subsequent repeat review of the CTSA hub websites will be conducted to determine evolving trends. RESULTS/ANTICIPATED RESULTS: A total of 40 responded to the survey from 59 CTSA hubs. Survey results are being analyzed. Website review data are being tabulated and the subsequent review of websites will be collected in February. Those findings are pending and will include a comparison of prior findings. 42% of CRP hubs list CRP training within the CTSA hub website. Required onboarding training (beyond CITI certificates) is revealed for some hubs (15%). DISCUSSION/SIGNIFICANCE OF IMPACT: On our initial website review less than half of the CTSA hub websites list specific CRP training on their website. Many were hidden behind firewalls and could not be reviewed for content. The REDCap Survey will provide more granular descriptions of programs. Data from a second website review will be collected for comparison. Based on a preliminary re-review of sites, there is a suggestion of increasing CRP workforce development information. CTSAs are well-positioned to be a central hub for promoting educational excellence of the institutional workforce, for medical centers and in other venues where clinical research is performed.

OBJECTIVES/GOALS: Novel EEG source localization and functional connectivity will assess frontal and cerebellar activity as predictors of subsequent memory and executive functioning (EF) decline in healthy, asymptomatic older adults who carry the Apolipoprotein-E ε4 allele (ε4+), which conveys up to 12x increased risk for Alzheimers disease (AD). METHODS/STUDY POPULATION: Healthy, cognitively intact ε4+ older adults (n = 23; ages 65-89) completed neuropsychological testing (focus on memory and EF) and EEG at Time 1, returning an average of one year later for neuropsychological retesting. EEG data during successful stop-signal inhibitory control trials will be used for advanced source localization and functional connectivity, with a focus on frontal and cerebellar regions of interest (ROIs). Source analyses will focus on the N200 time window (~200-350ms) to assess conflict processing and P300 (~300-550ms) for performance evaluation. Connectivity analyses are frequency-based, and will focus on theta band connectivity to assess conflict processing and delta to assess performance evaluation. RESULTS/ANTICIPATED RESULTS: Using hierarchical linear regression models, the magnitude of source activation within ROIs and connectivity metrics between ROIs will be used to predict residualized change in memory and executive function metrics between Time 1 and Time 2. We anticipate that 1) greater, compensatory activation in frontal ROIs during the N200 window, and 2) less cerebellar activation during P300, will predict memory and executive decline over the retest interval. Decline will also be predicted by 3) greater inter-hemispheric frontal connectivity in the theta band (conflict processing) and 4) less frontal-cerebellar delta connectivity (performance evaluation). DISCUSSION/SIGNIFICANCE: At most, ~50% of ε4 carriers will develop AD. Thus, identifying which carriers will decline is crucial to enabling successful, early intervention. Cerebellar dysfunction and impaired connectivity may be among the earliest indicators of incipient AD. Cutting-edge cerebellar EEG may enable an accessible option for early discernment of AD risk.

OBJECTIVES/GOALS: The Translational Science Benefits Model offers an approach for evaluating the downstream health and social impact of research. Using the new Translating for Impact Toolkit of nine web-based tools, researchers can create free, secure accounts to plan, track, and demonstrate the impact of their work. METHODS/STUDY POPULATION: Development of the online toolkit includes 6 phases: 1) Review of existing tools, 2) Development of fillable PDF tool prototypes, 3) Pilot testing, 4) Development of web-based tools, 5) Usability testing, and 6) Refinement of web-based tools. First, we reviewed existing tools for measuring research impact. We then created prototypes of nine tools, published on the TSBM website, and pilot tested with researchers. Based on feedback and testing, we developed and launched web-based versions of the tools. We are currently conducting usability testing with researchers, which we will use to evaluate the ease-of-use and quality of the tools, identify areas for improvement, and refine the tools. RESULTS/ANTICIPATED RESULTS: Researchers can sign up for user accounts, create projects, invite collaborators and program administrators, and save progress as the complete the nine tools in the Translating for Impact Toolkit. The tools are divided into three steps: Plan (Roadmap to Impact, Benefits 2x2, Partner Mapper, and Team Manager), Track (Impact Tracker), and Demonstrate (Product Navigator, Case Study Builder, Impact Profile, and Dissemination Planner). The toolkit also includes a dashboard that provides a quick snapshot of translational impact for each project. The toolkit will help both individual translational scientists demonstrate the impact of their work and CTSA hubs evaluate impact of their projects. DISCUSSION/SIGNIFICANCE: The TSBM online toolkit is a free, secure, easy-to-use platform researchers can use to plan for, track, and demonstrate the impacts of their work. The toolkit provides a structured process that will help the next generation of scientists prioritize and promote translational impact in their work.

OBJECTIVES/GOALS: Research emphasizes the importance of play in early childhood to support social, emotional, and physical development. This study explores how the Prescription for Play (P4P) is executed in clinical contexts by analyzing implementation fidelity and contextualizing the adaptations, challenges, and facilitators to the program’s functionality. METHODS/STUDY POPULATION: This project is an ongoing multi-site case study. At the time of study completion in December of 2023, there will be over 40 clinical observations of pediatric well-child check (WCC) visits completed across 7 Federally Qualified Health Centers (FQHC) participating P4P, a play promotion program wherein providers discuss the importance of play in WCC visits and provide a free play kit. All visits are with children 18-36 months old, with a broad demographic spread across sites. Observations are recorded through a guided observation protocol informed by a standard implementation fidelity framework, conducted by 5 researchers. Through inductive thematic analysis, this study will analyze observations of WCC visits to understand the ways providers engage with P4P across sociocultural contexts within FQHCs. RESULTS/ANTICIPATED RESULTS: Preliminary analysis of clinic observations (N = 30) indicates the degree of implementation fidelity varies across sites, with particular variances between WCC visits conducted in English versus non-English languages (NEL). In NEL visits, there were discrepancies among indicators of quality of delivery and participant responsiveness. NEL visits were less likely to have the provider model play with the caregiver and far less likely to open the play kit given to the family. Providers in NEL visits were also less likely to discuss certain benefits of play like brain development and reduced screen time. Across all observations, providers “prescribed play” approximately half the time. As more observations are conducted, researchers anticipate seeing continued differences between English and NEL visits. DISCUSSION/SIGNIFICANCE: From preliminary analysis, discrepancies in implementation fidelity indicate the P4P intervention may require adaptation and additional training related to how to prescribe and discuss play in WCC visits conducted in NEL visits. Additionally, this study elucidates the impact language can have on the fidelity of clinical interventions.

OBJECTIVES/GOALS: This project will design, develop and pilot-test a Spanish-mobile application for breast cancer patients. It is intended that patients will improve treatment adherence, have a personal information storage system for their follow-up healthcare and can have greater control in the processes of diagnosis, treatment, education, and recovery. METHODS/STUDY POPULATION: In the first phase, a team of undergraduate engineering students was recruited to work on the areas of: Front End, Database Development, Security Protocols, and Integration. Team Venus will provide the cancer-related content. In the second phase, the engineering team will identify bugs, usability, satisfaction, and workability of the application among a pilot convenience sample of 20 participants from four cancer clinics and the community. The inclusion criteria are being > 21 years old and having a cell phone. The exclusion criterion is not knowing how/being able to use a mobile application. Participants will complete a questionnaire on sociodemographic data, mobile phone and application use, efficacy, usability, and security. Univariate and bivariate statistical analysis will be performed using SPSS. RESULTS/ANTICIPATED RESULTS: The researchers are defining specific visual and content features they would like the application to have. They are also searching and collecting reliable information about cancer from primary sources to incorporate into the tools available to the application’s users. In addition, they have also begun to identify potential cancer treatment facilities and medical personnel to assist in the recruitment of patients for pilot-testing the application. The researchers have divided them into geographical areas in Puerto Rico and each Team Venus member will approach and orient patients and medical personnel about the project. DISCUSSION/SIGNIFICANCE: It is intended that patients will improve treatment adherence, have a storage system for their follow-up healthcare and greater control in their processes of diagnosis, treatment, education, and recovery. Future goals include use of the database feature by researchers and expanding the testing of the application with a larger patient-base.

OBJECTIVES/GOALS: This project examines the initial feasibility, acceptability, and appropriateness of using population-health data and geospatial information systems (GIS) technology to guide opioid overdose prevention in a Black community facing increasing racial inequities in fatal overdose since 2016. METHODS/STUDY POPULATION: We applied a mixed-methods approach, using GIS to examine hot spots of fatal opioid overdose among Black, adult decedents in St. Louis City and County, Missouri, derived from cause-of-death data from the Medical Examiner’s Offices from 2016-2021 (N = 1,612). Hot spot analysis was conducting using the Getis-Ord Gi* statistic. We then conducted a focus group with community-based providers in Black neighborhoods (N = 8) to assess the acceptability and appropriateness of mapping overdose data and to determine the utility of GIS for guiding decision-making in outreach activities aimed at overdose prevention. The focus group was recorded and transcribed for analysis to extract themes. RESULTS/ANTICIPATED RESULTS: Among 340 census tracts in the St. Louis region, 41 were statistically significant hot spots, indicating high clustering of fatal overdose. All hot spots were clustered together in North St. Louis, an area characterized by high racial and income segregation. Focus group participants reported potential pitfalls of GIS, including data inaccuracy for Black individuals and unwanted attention in hot spot areas (e.g., increased policing). Potential benefits included improved care coordination between organizations, assessing effectiveness of outreach interventions, and improving community-level awareness. Hyper-local neighborhood rather than regional maps were perceived as useful for outreach. DISCUSSION/SIGNIFICANCE: Artifacts of structural racism that drive Black opioid overdose deaths limit the utility of GIS to guide outreach in communities facing this systemic disinvestment. However, neighborhood-level GIS may serve as a tool for communities to build awareness and collaboration toward overdose prevention.

OBJECTIVES/SPECIFIC AIMS: N/A. METHODS/STUDY POPULATION: N/A. RESULTS/ANTICIPATED RESULTS: N/A. DISCUSSION/SIGNIFICANCE OF IMPACT: There is an increasing need to foster cross-disciplinary research to address complex problems within healthcare. The Sinai Team-based Translational Education Program: the URM Propeller (STTEP-UP) is a NCATS funded program through the Icahn School of Medicine at Mount Sinai. Its goal is to facilitate URM post-doctoral trainees becoming innovative leaders in clinical and translational research. The program includes a team-based research component, where fellows collaborate on a project. This year, disciplines represented by the four fellows include Cardiology, Psychiatry, Neurology, and Pediatrics. Identifying a clinical question and designing an investigation was facilitated by group brainstorming meetings with program mentors. Fellows designed a project to identify medical testing and prescribing that were not clinically indicated throughout the healthcare system, with the goal of exploring whether an intervention, including provider education, could reduce ordering practices. In addition to regular in-person meetings, a licensed virtual learning environment and free web-based sharing platform were used to foster collaboration. Challenges faced throughout this process, included fellows struggling to find protected time, difficulties accessing broad sets of data across the healthcare system, and overcoming administrative barriers between departments. Strengths of this approach, included fellows learning new research strategies and feeling a deeper sense of commonality with their peers. Overall, this experience supports the idea that cross-disciplinary research improves the collaboration and education of emerging researchers. However, addressing logistical and systems-based barriers may better facilitate this education and research.

OBJECTIVES/SPECIFIC AIMS: Endogenous RT (eRT) is necessary for the function of retrotransposons, elements that replicate via an RNA intermediate. One source of eRT activity is long interspersed elements (LINE). LINEs, of which there are several subgroups (L1, L2, L3), are retrotransposons that regulate cellular growth and gene expression. Given their diverse and important roles, we hypothesized that L1 elements regulate functional responses in megakaryocytes and platelets; a concept not yet examined in the field. METHODS/STUDY POPULATION: To study eRT in human platelets we used RT activity assays, PCR, and Western blot approaches. Furthermore, we used an RT-inhibitor to dissect the function of eRT, analyzed RT-dependent protein synthetic capacity, and immunoprecipitated RNA-DNA hybrids. RNA-DNA hybrids were also detected by means of ICC and automated analysis using CellProfiler software. RNA-DNA hybrids were validated by PCR and eRT regulated synthesis of target proteins was analyzed using autoradiography and Western blot techniques. Platelets from patients with HIV+ were examined in parallel. RESULTS/ANTICIPATED RESULTS: We identified that highly purified, isolated platelets from healthy subjects possess eRT activity. eRT activity was blocked with the non-nucleoside RT inhibitor nevirapine at concentrations within the therapeutic drug range. L1 elements are bicistronic, containing 2 open reading frames (ORFs), ORF1 and ORF2. Thus, we next identified that human platelets express full-length L1 mRNA containing ORF1 and ORF2. In human platelets, eRT activity was localized to L1 protein containing ribonucleo particles. Platelet eRT reverse transcribed exogenous RNAs, a process inhibited by nevirapine, acting in trans using the 3′-UTR of exogenous mRNAs as a template. To dissect the function of eRT in platelets, we next examined cytoskeletal and protein synthetic events in the presence or absence of nevirapine. Inhibition of eRT in isolated platelets led to characteristically beaded platelets in appearance, strongly resembling bone marrow proplatelets. Parallel increases in platelet reactivity were also observed. As these changes occurred over hours, not minutes, we hypothesized that inhibition of eRT would affect platelet protein synthetic events. Consistent with this, RT inhibition resulted in upregulation of global platelet protein synthesis. We validated upregulation of the synthesis of specific proteins (mitofilin, p-selectin, and L26—a component of the 60S ribosomal subunit essential for mRNA translation). RNA-DNA hybrids, noncanonical nucleic acid structures that regulate gene expression, are enriched in regions where L1 is abundant. RNA-DNA hybrids were present in platelets and expression confirmed via differential digestion of RNAs (eg, with RNase A and RNAse I). Next-generation sequencing of pulled down (eg, immunoprecipitated) platelet RNA-DNA hybrids identified numerous differentially expressed transcripts and we focused on MAP1LC3B (LC3B), a primary regulator of autophagy. Hybrid sequencing results for LC3B were validated using qPCR and we confirmed that LC3B RNA binds to L1-encoded RNA binding protein. Platelets treated with nevirapine had increased total LC3B protein expression. As RT inhibition is an important mechanism to control HIV infection, we examined platelet morphology, activation, and LC3B expression in platelets from HIV+ subjects treated with nevirapine. HIV+ patients treated with RT inhibitors had higher numbers of platelets that were beaded in appearance at baseline, increased platelet reactivity, and differential LC3B expression compared with healthy controls. DISCUSSION/SIGNIFICANCE OF IMPACT: Taken together, these results demonstrate that platelets possess eRT activity that regulates platelet morphology, platelet hyperreactivity, and protein synthetic events. We postulate that eRT activity in platelets may be a new post-transcriptional regulatory checkpoint. Moreover, our findings have implications in HIV+ patients treated with RT inhibitors, where off-target effects may contribute to platelet activation and an increased risk of thrombosis.

OBJECTIVES/GOALS: By establishing a youth-centric web-app as a central hub of information and inspiration in an attempt to engage a young demographic, this project aims to increase community awareness and reduce misconceptions surrounding clinical trials, in hopes of fairly representing marginalized communities among future clinical trial participants. METHODS/STUDY POPULATION: We designed a children’s web-app to host a collection of child-friendly educational materials (such as picture books, games, and age-appropriate articles about advances in clinical research) explaining clinical research and its process. An emphasis was put on ensuring the web-app and its contents were understandable and appealing to children. The effectiveness of this tool will be tested through a focus group study. Children ages 7-10 will be given a preliminary survey measuring their knowledge and opinions about clinical research, and then given time to explore the web-app. Afterwards, they will be given a secondary survey to gauge their acquired knowledge from the website and asked about their opinion on the design and usability of the web-app and its materials, as well as how likely they were to revisit the site. RESULTS/ANTICIPATED RESULTS: We anticipate a very positive response from the children regarding the design and usability of the web-app and its materials. By using an adolescent-focused design methodology at every step of the design process, we will ensure that all materials are attractive and engaging to our younger target audience. Exposing children to accessible information about clinical trials at a young age allows us to build their trust in the research process prior to the possible internalization and acceptance of cultural misconceptions. Over time, we hope to see a change in attitudes toward clinical research as well as increased participation, whether from under-represented groups or a younger demographic, and positively contribute to T3 and onwards in the translational continuum. DISCUSSION/SIGNIFICANCE: In a rapidly changing world, the best approach to making change is through targeting the younger demographic, the leaders of tomorrow. Our project will allow adolescents to foster a more well-rounded opinion of clinical research, increasing their participation and better paving a more positively received future for translational science as a whole.