OBJECTIVES/GOALS: Computational pathology is an emerging discipline that resides at the intersection of engineering, computer science, and pathology. There is a growing need to develop innovative pedagogical approaches to train future computational pathologists who have diverse educational backgrounds. METHODS/STUDY POPULATION: Our work proposes an iterative approach toward teaching master’s and Ph.D. students from various backgrounds, such as electrical engineering, biomedical engineering, and cell biology the basics of cell-type identification. This approach is grounded in the active learning framework to allow for observation, reflection, and independent application. The learners are trained by a team of an electrical engineer and pathologist and provided with eight images containing a glomerulus. They must then classify nuclei in each of the glomeruli as either a podocyte (blue), endothelial cell (green), or mesangial cell (red). RESULTS/ANTICIPATED RESULTS: A simple web application was built to calculate agreement, measured using Cohen’s kappa, between annotators for both individual glomeruli and across all eight images. Automating the process of providing feedback from an expert renal pathologist to the learner allows for learners to quickly determine where they can improve. After initial training, agreement scores for cells scored by both the learner and the expert were high (0.75), however, when including cells not scored by both the agreement was relatively low (0.45). This indicates that learners needed more instruction on identifying unique cells within each image. This low-stakes approach encourages exploratory and generative learning. DISCUSSION/SIGNIFICANCE: Computation medical sciences require interdisciplinary training methods. We report on a robust approach for team-based mentoring and skill development. Future implementations will include undergraduate learners and provide opportunities for graduate students to engage in near-peer mentoring.

ABSTRACT IMPACT: We developed a novel semi-automated computational platform to quantify spatial characteristics of preserved peripheral visual field loci in retinitis pigmentosa. This work may inform future research on therapies to prevent visual field loss in retinitis pigmentosa and related diseases. OBJECTIVES/GOALS: Retinitis pigmentosa (RP) causes progressive and severe peripheral visual field (pVF) loss in some but not all patients. The characteristics of pVF in RP are incompletely understood. We developed a novel semi-automated computational platform to quantify the spatial characteristics of pVF. METHODS/STUDY POPULATION: We analyzed preserved pVF size and location in both eyes of RP patients using the Goldmann V4e isopter. We developed a custom algorithm in MATLAB to align and average the binary V4e isopter segmentations, and generated a two-dimensional probability map of the spatial distribution of preserved pVF loci along the radial and circumferential dimensions. To adjust for disease duration, cases were categorized by the time from self-reported symptom onset. Probability maps of pVF preservation were generated for categories of disease duration using unsupervised K-means clustering. Analyzing cases with longitudinal data, we identified loci of stable pVF over time. RESULTS/ANTICIPATED RESULTS: A total of 152 patients were included (N=304 eyes). The mean age was 46.7 years and 49.3% were male. Disease duration was categorized as <20 years (N=72, 47.4%), 20-40 years (N=60, 39.5%), or >40 years (N=20, 13.2%). Longitudinal data (3.2 -5.7 years of follow-up) were available in 65 patients (42.8%). Probability plots of preserved pVF loci in the cross-sectional dataset showed that the median percentage of preserved pVF loci were located between 50 ºand 80 ºeccentricity and between the 30 ºto 50 ºmeridians, with highly concordant inter-ocular symmetry. Probability plots in the longitudinal dataset showed that inferotemporal pVF loci were most likely to be preserved over time. DISCUSSION/SIGNIFICANCE OF FINDINGS: Semi-automated quantification of pVF loci is a useful platform to analyze spatial characteristics of the visual field in RP. Certain portions of the pVF may be relatively resistant to functional decline. Understanding the molecular basis of pVF resilience will inform further research on RP therapy.

OBJECTIVES/GOALS: An estimated 87% of torture survivors, or 27 million people globally, suffer from chronic pain such as brachial plexopathy from upper extremity suspension or lumbosacral plexus injury from leg hyperextension. However, a vast majority of pain is undetected by evaluators due to a lack of diagnostic tools and confounding psychiatric illness. This diagnostic gap results in exclusive psychological treatment rather than multimodal therapies, substantially limiting rehabilitation, placing vulnerable individuals at higher risk of drug abuse, and increasing healthcare expenditures. We hypothesized that the novel application in torture survivors of a validated pain screen can supplement the UNIP and improve its sensitivity for pain from approximately 15% to 90%, as compared to the reference standard. METHODS/STUDY POPULATION: In this prospective, blind comparison to gold standard study, 25 patients who survived torture by World Medical Association definition first received the standard evaluation protocol for torture survivors (United Nations Istanbul Protocol, UNIP) by a trained evaluator, and subsequently received a validated pain screen (Brief Pain Inventory Short Form, BPISF) followed by a non-invasive examination by a pain specialist physician (reference standard). The primary outcome was the diagnostic and treatment capability of the standard protocol (index test) versus the validated pain screen (BPISF), as compared to the reference standard. RESULTS/ANTICIPATED RESULTS: Providers using only the UNIP detected and treated pain in a maximum of 16% of patients as compared to 85% of patients who were diagnosed with pain by the reference standard. When employed, the validated pain screen had a sensitivity of 100% [95% CI: 72% - 100%] and a negative predictive value of 100%, as compared to a sensitivity of 24% [95% CI: 8% - 50%] and negative predictive value of 19% by the index test. The difference in the sensitivity of the UNIP as compared to the BPISF was significant, with p < 0.001. DISCUSSION/SIGNIFICANCE OF IMPACT : These data indicate that the current global standard assessment of torture survivors, the United Nations Istanbul Protocol, should be supplemented by the use of a validated pain screen to increase the accuracy of chronic pain diagnosis. This would change the standard medical assessment of 70.8 million people globally, a number that continues to rise by nearly 45,000 people each day. ClinicalTrials.gov protocol number NCT03018782.

OBJECTIVES/SPECIFIC AIMS: In June 2016, the FDA cautioned against the use of SGLT-2 inhibitors because of increased risk of acute kidney injury (AKI) after 101 cases of AKI were reported between March 2013 and October 2015. This study seeks to determine risk of AKI associated with SGLT-2 inhibitors in a large cohort of type 2 diabetic patients. METHODS/STUDY POPULATION: Retrospective cohort study including SGLT-2 inhibitor users and nonusers in the Mount Sinai Chronic Kidney Disease Registry between January 2013 and September 2016. SGLT-2 inhibitor users and nonusers were type 2 diabetics with new SGLT-2 inhibitor prescription after January 2013 and an outpatient visit between 2013 and 2015, respectively. Subjects were propensity matched by nearest neighbor method based on demographics, comorbidities, laboratory values, medications, estimated glomerular filtration rate (eGFR), and length of follow-up. The primary end point was AKI (defined by KDIGO laboratory algorithm) occurring during the follow-up period. RESULTS/ANTICIPATED RESULTS: In total, 372 SGLT-2 inhibitor users [mean age 63 years; 205 (55%) men] and 372 [mean age 63; 194 (52%) men] nonusers were included in the primary analysis. Proportions of AKI events defined by KDIGO criteria in users and nonusers were 4.0% and 10.0%, respectively. Adjusted odds ratio for AKI was 1.00 (95% CI, 0.28–2.62). Median peak serum creatinine measurements during AKI events for user and nonuser groups were 1.60 (IQR 1.36–1.78) and 1.88 (IQR 1.55–2.44) (p=0.02), respectively. Sensitivity analyses yielded similar results. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings suggest that there is no evidence of increased odds of AKI in SGLT-2 inhibitor users compared with propensity-matched nonusers with type 2 diabetes.

OBJECTIVES/GOALS: Incorporating health equity, diversity, and inclusion (HEDI) development during community advisory board meetings is essential for ensuring that perspectives of all community members are considered, that health research is centered on the experiences of historically marginalized groups, and organizational strategies align with the community. METHODS/STUDY POPULATION: All IN for Health is a public engagement program that promotes health and research literacy, seeking to increase the public’s understanding of the role and value of research in improving health. It is guided by an active CAB providing advice on strategic directions, feedback to all efforts, contributing ideas, priorities, and most importantly, accountability. In 2023, ALL IN for Health started to incorporate HEDI professional development during staff team meetings and most recently, into our quarterly CAB meetings. Initially, the CAB was asked to provide feedback about talks and potential presentations related to health-research, sponsored, and shared by All IN for Health. During this exercise, board members were asked to provide HEDI topics of interest. Their responses informed a plan for team and board members. RESULTS/ANTICIPATED RESULTS: Some of the topics suggested by board members include: understanding health equity in relation to research studies and protocols, and topics specific to health research such as: health care access for rural areas and vulnerable populations, culture-based attitudes and beliefs and how they impact decisions related to health care, being aware of limitations certain communities have and what they may not have access to. The initiative was positively received and unanimously adopted. We hope to introduce the HEDI integration model for CABs at this conference. (Conceptual Model attached). DISCUSSION/SIGNIFICANCE: It is important to learn and grow alongside our community members. Such practice is bound to sustain partnerships that promote health equity, and exemplify meaningful community engagement, bidirectional learning, and a shared leadership model. By consistently incorporating and prioritizing HEDI, HAB can contribute to more equitable initiatives.

OBJECTIVES/SPECIFIC AIMS: The objective of this study is to measure the association of CYP2C19 (*1-*8,*17), ABCB1(C3435T; rs1045642), PON1 (p.Q192R; rs662), and B4GALT2 (c.909 C>T and c.366 G>C) gene polymorphisms in the Caribbean Hispanic population with major adverse cardiovascular events (MACE). METHODS/STUDY POPULATION: Patients of Caribbean Hispanic ethnicity from all geographic regions of the Island of Puerto Rico, male and female, aged >21 will be recruited. Cases will consist of patients receiving a daily clopidogrel dose of 75 mg following acute coronary syndrome (ACS) who experience a MACE within the first year of treatment. Control study patients must have received clopidogrel 75 mg daily for a minimum of 1 year without experiencing MACE. Genomic DNA samples will be genotyped to determine the frequency distribution of major CYP2C19, ABCB1, PON1, and B4GALT2 gene polymorphisms. Observed frequencies will be compared with other reported populations. An association study will be performed between genetic variables and MACE and a multivariable logistic regression model (additive) will be constructed. RESULTS/ANTICIPATED RESULTS: We anticipate finding a significant association between major genetic determinants of clopidogrel response and MACE where cases with MACE will carry higher frequency of CYP2C19, ABCB1, PON1, and B4GALT2. DISCUSSION/SIGNIFICANCE OF IMPACT: As the range of multiloci allelic combinations in admixed Caribbean Hispanics is higher than in other populations due to its unique 500-year history of genomic admixture, a wide spectrum of genetic variances is expected to be present in the study population. Determining the prevalence and effect of CYP2C19, ABCB1, PON1, and B4GALT2 polymorphisms holds the potential to personalize anti-platelet treatment for Caribbean Hispanic patients requiring treatment after ACS.

OBJECTIVES/GOALS: The design and phased roll-out of a CTSA competency-based longitudinal Clinical and Translational Research (CTR) curriculum pathway that can be integrated into the training of health sciences professional degree programs at three regional institutions. The outcome will be an increased number of health science professionals participating in CTR. METHODS/STUDY POPULATION: Based on an environmental scan, student surveys to assess interest, and feedback from regional stakeholders, a CTR pathway program was developed. The pathway curriculum will be grounded in 6 key focus areas derived from the Core Competencies in Clinical and Translational Research and prioritized based on regional need. The CTR pathway is currently being developed for the University of Washington School of Medicine, with additional implementations at the Washington State University School of Pharmacy and the Montana State University School of Nursing. Students will complete training modules and a mentored research project that is integrated into their specific course of study. RESULTS/ANTICIPATED RESULTS: In addition to the initial assessments and curriculum, an Advisory Committee will be established. Mentors, site leads, and research project partnerships across the region will be identified. Modifications will be made according to the local needs at both Montana State University and Washington State University. After the pilot launch and roll-out, the pathway curriculum will be adapted for other disciplines based on input from content experts and pathway evaluation data. Student retention in CTR fields will be tracked, with a goal to increase the number of CTR investigators and professionals across WWAMI in the next decade. CTR pathway processes and training resources will be shared with the CTSA consortium and other health sciences professional training sites. DISCUSSION/SIGNIFICANCE: Development of the CTR workforce is a priority to increase national capabilities in clinical and translational science. Building on a recognized need for targeted and longitudinal engagement, a CTR pathway is being established for health sciences students in the WWAMI region.

OBJECTIVES/GOALS: The primarygoal is to understand the challenges and barriers associated with the procurement of innovative technologies.Specifically, our research will answer the following question: what are the minimal requirements for a startup’s solution to beprocuredby anOntariohealthcare institution? METHODS/STUDY POPULATION: Participants will include procurement professionals at startups, healthcare institutions, and procurement facilitating agencies. Semi-structured interviews will be conducted in order to understand different procurement pathways and the possible procurement related gaps or barriers that startups face. Through qualitative ethnographic methods, participant interviews will characterize existing relationships and examine the rationale behind startup procurement decision-making. Data collection will include recordings, verbatim transcripts, and researcher field notes. Through inductive qualitative analysis, the data will be examined to build an intervention to assist in startup procurement. RESULTS/ANTICIPATED RESULTS: Our investigation will yield insight into expectations between hospital procurement requirements and startup procurement. The qualitative analysis will identify targets for engagement, and appropriate actors that can bridge gaps. Our results will identify pathways for procurement and the minimal procurement requirements to aid startup procurement planning. Our research will support innovators by delivering an intervention that will enable easier implementation of market ready solutions in a Canadian context. In line with principles from the National Center for Advancing Translational Sciences, this research can be used towards enhancing efficiency, speed of translation, and innovation. DISCUSSION/SIGNIFICANCE: We will contextualize the needs of start-ups and empower them to understand their procurement ecosystem. Facilitating better navigation of the procurement space allows for innovators to present solutions that healthcare organizations can adopt, resulting in improved clinical and patient outcomes.

OBJECTIVES/SPECIFIC AIMS: We investigated the association between relationship power imbalance (which can have a negative impact on HIV prevention) and male partner HIV testing, using baseline data from a HIV self-testing trial in 3 antenatal clinics in central Uganda. METHODS/STUDY POPULATION: Pregnant women with HIV-male partners were recruited and randomized by day into standard of care or intervention (HIV self-testing kits). Analyses were performed in SAS 9.4, with χ2 tests and p<0.05 for significance. RESULTS/ANTICIPATED RESULTS: In total, 1514 women were recruited (737 standard of care, 777 intervention). Overall, 39.6% of male partners had previously tested for HIV. Among women <26, contributions to expenses differed by partner testing (overall p<0.001, 47.6% of women whose partners tested made no contribution vs. 63.2% of women whose partners did not test). Relationship status differed by partner testing (overall p=0.02, 12.4% of women whose partners tested showed a sometimes difficult relationship vs. 5.7% of women whose partners did not test). Among women 26+, decision making for family visits differed by partner testing (overall p=0.005, 52.9% of women made joint decisions with partners who tested vs. 36.5% whose partners did not test). DISCUSSION/SIGNIFICANCE OF IMPACT: Higher relationship power balance was associated with higher HIV testing among male partners when measured by contribution to expenses and decision making for family visits, but not relationship status. Relationship power balance should be considered when counseling women and men to increase HIV testing.

OBJECTIVES/GOALS: In November 2020, Tufts CTSI launched a new, upgraded learning management platform for disseminating self-paced online professional education for the CTR workforce. This poster will present a case study of the first two full years of Tufts CTSI I LEARN usage to illustrate how a local and national CTR workforce engages in free online training. METHODS/STUDY POPULATION: This study will employ a retrospective analysis of 2021 and 2022 learner usage data from the Tufts CTSI I LEARN platform. The population are individuals who have engaged in one or more courses over the specified time period, where engagement is defined as having viewed at least one content item in a course. Descriptive statistics will be used to show total engagement and unique learners each year by course, popular courses by year, as well as geographical distribution of learners across both years. Two sample t-tests will be used to compare the mean percentage of course completion and mean time spent in courses for 2021 and 2022. The Wilcoxon Rank Sum test will be used to compare median courses taken per learner each year. Finally, descriptive statistics will be used to show returning learners vs new learners from 2021 to 2022. RESULTS/ANTICIPATED RESULTS: Data from 2021 show 723 learners (436 unique) engaged in 49 courses across the year. Course engagement ranged from a max of 135 to minimum of 1 learner (Mean = 14.8, Median = 6). Mean percent of course content viewed was 69.4 (Median = 63.7%) and mean time spent on course content was 41.3 minutes (Median = 33.6). Preliminary data from Q1-Q3 of 2022 find 557 learners (328 unique) have engaged in 69 courses. Based on this trend, it is anticipated that 2022 will match or exceed 2021 engagement levels. Recently launched courses are expected to have higher average engagement than legacy courses ported from the previous version of the platform. DISCUSSION/SIGNIFICANCE: Results from this evaluation will suggest strategies to enhance engagement in current courses and identify opportunities for future course development. Increasing engagement in self-paced CTR education is an important complement to formal workforce development and training programs.

OBJECTIVES/GOALS: To engage community partners in full spectrum of translational sciences participants will: 1. Develop community-based research projects. 2. Evaluate pilot awards submitted to the Community Advisory Board for funding. 3. Actively participate in the research cores at the CTSC METHODS/STUDY POPULATION: The Academy workshop curriculum supports an in-depth examination of the translational research process, introduction to the CTSC Community Advisory Board; as well as to community based participatory research; grant evaluation process, and the need for active community involvement in various cores, and clinical trials. Workshops presented by tenured professionals and community leaders with multi-media out of class assignments. Scheduled sessions were 1 hour, and 15 minutes. Exams given post class session with a mean score of 22.8/30 for session 1 and a mean score of 23.80/30 for session 2. RESULTS/ANTICIPATED RESULTS: The Community Research Academy (CRA) has thus far held for two sessions. Of the 45 enrollees, 20 completed the entire program. Of those, ten have joined the CAB board; and three have actively participated in the pilot award review process. One of the CRA graduates will be publishing her photo novella assignment in Health Affairs. One person changed careers,: and several people have taken all the classes for a second time. Many participants originally came from our faith-based connections. Now word of mouth is expanding the program. DISCUSSION/SIGNIFICANCE: Community engagement ensures that the fresh voices of diverse populations are involved in translational science. Their input ultimately leads to creating novel clinical innovations; such community-driven ingenuity which addresses the deeply felt needs of those communities.

OBJECTIVES/GOALS: We investigated hemodynamic measures in young black and white adults below the age of 50 years to identify mechanisms that may predispose blacks to more CVE. METHODS/STUDY POPULATION: We recruited 276 young blacks and white adults in Jackson, MS (mean age: 33 ± 9 years; 70% women; 57% Black;). Participants had clinical and vascular tonometry parameters obtained. Vascular measures included carotid femoral pulse wave velocity, central and peripheral pulse pressure (CPP and PPP), characteristic impedance (Zc), augmentation index, and forward pressure wave. Race-specific characteristic means and proportions were tested using Student’s T- or Chi-square tests. After further sex stratification, significant variation in characteristics among sex within race were tested using nested ANOVA. RESULTS/ANTICIPATED RESULTS: Characteristics of the study group stratified by race were found to be similar. Vascular measures stratified by race revealed blacks to have significantly higher Zc (p = 0.03) and PPP (p = 0.03) than whites. DISCUSSION/SIGNIFICANCE OF IMPACT: In this study of vascular hemodynamics in young black and white participants we found differences in Zc and PPP. Findings suggest that differing relations between proximal aortic diameter and wall stiffness may contribute to the racial disparity in CVE in adults. This finding could offer an explanation to the beneficial effects of treatment modalities that target aortic stiffness in blacks.

OBJECTIVES/SPECIFIC AIMS: Triple-negative breast cancer (TNBC) accounts for one-fifth of the breast cancer patient population. The heterogeneous nature of TNBC and lack of options for targeted therapy make its treatment a constant challenge. The co-deficiency of tumor suppressors p53 and ARF is a significant genetic signature enriched in TNBC, but it is not yet clear how TNBC is regulated by this genetic alteration. METHODS/STUDY POPULATION: To answer this question, we established p53/ARF-defective murine embryonic fibroblast (MEF) to study the molecular and phenotypic consequences in vitro. Moreover, transgenic mice were generated to investigate the effect of p53/ARF deficiency on mammary tumor development in vivo. RESULTS/ANTICIPATED RESULTS: Increased transformation capability was observed in p53/ARF-defective cells, and formation of aggressive mammary tumors was also seen in p53-/-ARF-/- mice. RNA-editing enzyme ADAR1 was identified as a potential mediator for the elevated oncogenic potential. Interestingly, we found that the overexpression of ADAR1 is also prevalent in human TNBC cell lines and patient specimen. Using short hairpin RNA (shRNA) to reduce ADAR1 expression abrogated the oncogenic potential of human TNBC cell lines, while non-TNBC cells are less susceptible. Different levels of RNA editing of known ADAR1 targets were detected in shRNA-treated human TNBC cell lines, suggesting that ADAR1-mediated RNA editing contributes to TNBC pathogenesis. DISCUSSION/SIGNIFICANCE OF IMPACT: These results indicate critical roles played by the tumor suppressors p53 and ARF in the pathogenesis of TNBC, partially through affecting ADAR1-mediated RNA editing. Further understanding of this pathway could shed light on potential vulnerabilities of TNBC and inform the development of personalized therapies based on patients’ genetic signiatures.

ABSTRACT IMPACT: This change will improve primary care physicians and pediatrics ability to identify, intervene and prevent obesity related renal damage in the vulnerable population of young adults OBJECTIVES/GOALS: Obesity related glomerulopathy has a reversible stage manifested as hyperfiltration. Early intervention depends on the ability to identify hyperfiltration. Hyperfiltration prevalence is underestimated using the currently recommended formula We investigated whether calculating BSA-adjusted GFR will more readily identify hyperfiltration. METHODS/STUDY POPULATION: We extracted data from a large urban, multi-institutional Electronic Health Records (EHR) clinical data research network to construct an EHR data base of 60,549 women and girls ages 12-21 years from the New York metropolitan area. EGFR was calculated in two ways, 1) according to age appropriate formula, and 2) according to age appropriate formula and adjusted to body surface area (BSA). BMI-for-age values were classified according to the World Health Organization schema and grouped according to the CDC definitions. BSA was calculated according to the Du-Bois formula. Hyperfiltration was defined by a threshold of 135ml/min. The Bland Altman method assessed the agreement between formulas across the different BMI groups. RESULTS/ANTICIPATED RESULTS: Serum creatinine values were similar across different BMI groups. Comparing eGFR values, hyperfiltration rates were similar across BMI groups, ranging between 4%-6.6%. BSA-adjusted GFR was different across BMI groups: hyperfiltration rates were 0.81% for the underweight group, 2.56% for the normal weight, 12.18% for the overweight and 39% in the obese group. This trend of hyperfiltration paralleled the the rise in urine creatinine across BMI groups. DISCUSSION/SIGNIFICANCE OF FINDINGS: BSA-adjusted GFR more sensitively detects hyperfiltration due to obesity than does eGFR. Calculating BSA-adjusted GFR will improve primary care and pediatric physicians’ ability to identify, intervene and prevent early ORG. Changes in body composition may account for the increasing discordance between BSA-adjusted and eGFR as BMI rises.

Story is the oldest known way of sharing knowledge and information and engages us in our collective humanity. In research settings, story brings meaning to complex ideas, making them feel palpable and connects us with our audience. Historically, the disciplines that take a research interest in the importance of narrative have been largely in fields like the philosophy of science/medicine, medical humanities, and sociology though story is “always already” a part of scientific research. Humanities have gained traction in medical and science education, and researchers are seeking such curricula to communicate more effectively with the public and their students. We believe that story is an effective tool to enable CTS investigators to be effective educators and communicators of translational science. Story-based interdisciplinary pedagogy emphasizes an approach encouraging clinical researchers to keep the human story as the driving force of research design, dissemination, and application of research to diverse audiences. In this article, we provide backgrounds on successful programs that have used story in science communication and education as well as a tool researchers can use to incorporate the structure of story into their own work.

OBJECTIVES/GOALS: Alachua TranQuility was formed in early 2016 to address needs of transgender people in Alachua County, Florida and surrounding areas. Increasing awareness of gender identity issues created a growing population in need of additional health and social services to counter the negative health outcomes traditionally experienced by transgender people. METHODS/STUDY POPULATION: The group met monthly at UF HealthStreet, a CTSA community engagement program, where social and medical referrals and opportunities to participate in research were made available to attendees. Those attending included transgender and gender non-conforming individuals as well as family, friends and other allies. Outreach was conducted through social media, physical fliers, and tabling at community events. TranQuility partnered with local LGBTQ organizations to promote activities. Leadership was provided by a steering committee that changed over time, with feedback from attendees guiding planning. Simultaneous to TranQuilitys founding, a Youth Gender Clinic was established at UF Health, clearly demonstrating the need for services aimed at the transgender population. RESULTS/ANTICIPATED RESULTS: Between February 2016 and September 2019, attendance at meetings was logged per HealthStreet protocols. Age of attendees ranged from 10 to 75, and most were non-Hispanic whites. More than 300 individuals attended meetings across time, and many attended multiple meetings, with greater than 15% having attended five or more meetings. Attendance averaged around 30+ each month, with a high of 76. Educational programming was offered, such as information on hormone therapy and name/gender marker change, and a parent support group was formed to meet concurrently with the main group. Social opportunities were very popular, as a safe place for people to present as their authentic selves. The coronavirus pandemic curtailed in-person meetings at HealthStreet in 2020, but the group plans to return to this format in 2022. DISCUSSION/SIGNIFICANCE: TranQuility has become an established organization to which other groups provide referrals. Qualitatively it is clear that many people have been positively impacted. An ongoing discussion for the group, however, has been to improve outreach to transgender people of color, who typically suffer from the most discrimination and anti-trans violence.

OBJECTIVES/GOALS: This project focuses on investigating the potential of Simalikalactone D (SKD) as an anticancer agent, exploring the mechanisms underlying SKD’s induction of cell death, and assessing the impact of SKD on diverse breast cancer cell lines. Also, it Investigates the compound’s mechanisms of action beyond caspase 3-dependent pathways. METHODS/STUDY POPULATION: Three breast cancer cell lines were used: SKBR3, MDA-MB-231, and MDA-MB-468. Two triple-negative breast cancer cell lines are included to address cancer disparities across diverse ethnic backgrounds. Viability assays were conducted to determine half-maximal inhibitory concentrations (IC50). Caspase 3 activity assay was performed to evaluate apoptosis as a possible cell death pathway. Wound healing and colony formation assays are used to assess cell migration and clonogenic capacity. Proteomic analysis and phosphoarray analysis are planned for a deeper understanding of SKD’s anticancer properties, as well as testing for caspase 3 independent pathways. RESULTS/ANTICIPATED RESULTS: SKD demonstrated substantial cytotoxicity against all three breast cancer cell lines. IC50 values for SKBR3, MDA-MB-231, and MDA-MB-468 were 60.0 nM, 65.0 nM, and 116 nM, respectively. SKD induces cell death via caspase 3-independent pathways. Further experiments are needed to confirm and elucidate the molecular pathways being impacted. SKD inhibited cancer cell migration and clonogenic potential, suggesting it can reduce tumor growth and metastatic tendencies. DISCUSSION/SIGNIFICANCE: The study highlights SKD’s cytotoxicity across diverse breast cancer cell lines. It underscores the mechanism of action, a caspase 3 independent pathway. These findings hold promise for the development of innovative anticancer treatments and emphasize the importance of exploring varied cellular responses to mitigate global cancer disparities.