OBJECTIVES/SPECIFIC AIMS: This project aimed to offer Citizen Scientists basic knowledge about clinical and translational research, as well as introductory information to the topics a Citizen Scientist may work with at the University of Florida (UF). As part of that goal, several objectives were identified: (1) create a curriculum to operationalize training for new Citizen Scientists, (2) utilize best practices in educational research for curriculum design and implementation, (3) create a resource that can be freely available to other groups, and (4) implement the finished curriculum with Citizen Scientists at UF. METHODS/STUDY POPULATION: Working with an expert in the field of instructional design, an overall design plan was created and implemented. This included first conducting interviews with Citizen Scientists to determine the most appropriate format for the content, creating 10-minute videos with subject matter experts, and crafting learning assessments for each didactic video. Topics for the curriculum were conceptualized by gathering input from CTSI leaders, Citizen Scientists, and staff members, and learning objectives were created to help guide the content creation. The ADDIE (Analysis, Design, Development, Implementation, Evaluation) instructional design model was utilized to help guide the creation process, and this included a formative evaluation of the content, where assessment questions or videos were edited in response to Citizen Scientist feedback. It was important to both CTSI leadership and the Citizen Scientists themselves that the curriculum be widely accessible, so the curriculum was made as an open educational resource, meaning that it is available online for use by anyone with content that can be customized to specific programs or organizations. To implement the curriculum with UF Citizen Scientists, the materials were ported into Canvas, a widely utilized learning management system at UF. Participants were split into two groups, one group with Citizen Scientists already in the program (n=6) and a group with new Citizen Scientists (n=2). IRB approval to conduct this pilot test and share the results was obtained, and implementation ran from July 2017- January 2018. RESULTS/ANTICIPATED RESULTS: Data were taken from participant scores on the curriculum’s 15 didactic learning assessments, and while the number of attempts on assessments was not limited, only first-attempt results were analyzed. Veteran Citizen Scientists scored higher than the new Citizen Scientists, with an overall score across all assessments of 84% vs 74%. The existing Citizen Scientists performed better on most topics, however the newer Citizen Scientists outscored the veterans on two modules: Cultural Competency (90% vs 77%) and Biomedical Informatics and Big Data (80% vs 73%). The newer members also had fewer retakes on these two modules, with the newer members having only one retake for these two modules, compared to seven total retakes across both modules for the established members. Participants were also asked basic questions about learning comprehension, video quality, and assessment item clarity, in addition to offering narrative feedback. Participants across both groups seemed largely pleased with the curriculum, as indicated by results of the course evaluation. Most (75%) Citizen Scientists felt it was easy to understand the information in the video tutorials, while 75% of Citizen Scientists felt that the assessment items were comprehensive, and 62.50% felt that the assessment items matched the learning objectives. The new Citizen Scientists were far more likely to respond favorably to the video and assessment evaluation questions, with 95% of all responses marked as “agree” or “strongly agree”, compared to 57% of the responses from the established Citizen Scientists. DISCUSSION/SIGNIFICANCE OF IMPACT: The performance of new vs veteran Citizen Scientists underscores the need for introductory information on clinical research topics for new community stakeholders, as well as the need for ongoing refresher training. Though the existing Citizen Scientists had been working with these topics already, and some had experienced the modules already through the formative evaluation, some topics remained a challenge for participants, a notion reflected in their assessment scores. Clinical research, particularly translational research, can be difficult to understand, and when a stakeholder understands the fundamentals underlying the research with which they are assisting, communication barriers are eliminated, feedback is well-reasoned and actionable, and there is greater buy-in from stakeholders. Understanding the clinical research process helps community stakeholders better understand their contribution to research and offer critical feedback to aid in implementation of research findings in health-related settings.

Objectives/Goals: Research participants are not representative of our communities. To determine research perceptions, we are recruiting individuals who have and have not participated in studies. We aim to identify trends, capturing motivating factors, access barriers, and participants’ concerns to inform best practices to engage participants. Methods/Study Population: We developed a 25-item survey with a goal of engaging 500 individuals. Survey questions were developed based on published literature of research motivations and barriers, as well as focus groups conducted in our central Pennsylvania community. Survey questions consist of Likert-scale, multiple choice, and free text, with administration through REDCap. Potential survey participants will be recruited from the Penn State Volunteer Repository and through partnering clinical networks, including federally qualified health centers. We will also recruit individuals who have never participated in research to engage underrepresented groups via the Penn State Clinical and Translational Science Institute’s community partner network. Analyses will include Chi-squared tests and binomial logistic regression. Results/Anticipated Results: The primary dependent variable will be past participation in research, while the primary independent variables will be binary responses to questions relating to motivators and barriers to participation in research. Secondary analysis will stratify these comparisons by race or rurality with a subgroup analysis using the same methods as the primary analysis. Based on these analyses, we anticipate that those who have never participated in research will face greater and different barriers than those who have participated in research. We also anticipate that motivators will be similar between to the two groups. We anticipate finding differences in motivators and barriers among different races as well as differences when examining rurality versus non-rurality. Survey results will be available in January. Discussion/Significance of Impact: This will be one of the few studies in the literature focused on uncovering motivators and barriers to research participation. Understanding what motivates and prevents individuals from participating in research is one of the first steps to enhancing the research process so that people want, and are able, to participate in studies.

The U.S. Food and Drug Administration (FDA) recently harmonized its regulatory criteria for waivers and alterations of informed consent with the U.S. Common Rule. We conducted a systematic review of clinical trials in the two highest-impact clinical journals in the U.S. to better understand practice-changing trials that are being conducted in the U.S. without prospective written informed consent. One in ten trials in our sample were conducted without prospective written informed consent. Such trials accounted for 93.1% of patient-subjects in our sample, enrolling nearly 3 million individuals. Further guidance would support socially valuable research while ensuring appropriate oversight.

The Recruitment Innovation Center (RIC) has created a toolkit of novel strategies to engage potential participants in response to recruitment and retention challenges associated with COVID-19 studies. The toolkit contains pragmatic, generalizable resources to help research teams increase awareness of clinical trials and opportunities to participate; produce culturally sensitive and engaging recruitment materials; improve consent and return of results processes; and enhance recruitment of individuals from populations disproportionately impacted by COVID-19. This resource, the “RIC COVID-19 Recruitment and Retention Toolkit,” is available free online. We describe the toolkit and the community feedback used to author and curate this resource.

New education and training opportunities are critical for the development of a diverse and highly skilled translational science workforce. In this special communication, the authors consider how Narratives of Discovery, an initiative to interview leading scientists about the sources of their creativity, can serve as a novel translational science teaching tool. Reporting on a project to map translational science principles onto nine Narratives of Discovery conducted to date, the authors demonstrate how translational science principles are manifested in the career trajectories of these scientists and propose that the narratives can serve as a formative model for trainees. Findings from systematic coding of the Narratives of Discovery suggest that the narrative format is particularly well suited to highlight translational science principles not well-addressed by existing education opportunities, including what it means for scientists to be creative and innovative, use bold and rigorous approaches, and prioritize diversity, equity, inclusion, and accessibility. Offering excerpts from the published Narratives of Discovery and quotations from the scientists themselves, the authors aim to create space for continued conversation about how to best crystallize the concepts of translational science and advance translational science education and training initiatives.

OBJECTIVES/GOALS: Aim1: To develop a natural language processing (NLP) algorithm to effectively identify statin associated muscle symptoms (SAMS) in patients’electronic health records (EHRs). Aim2: To develop a machine learning model based on clinical features within EHRs that predict the likelihood of SAMS occurrences. METHODS/STUDY POPULATION: A retrospective cohort of adult patients initiated on statins within the Minnesota Fairview Healthcare System EHRs from 2010 to 2020 will be analyzed. NLP-PIER (Patient Information Extraction for Research) platform will be used to search and identify patients who developed SAMS after statin initiation. Manual annotation of clinical notes will be completed to validate the accuracy of identified SAMS cases. Then, a selection of clinical features within the EHRs will be input as predictors for machine learning algorithms development. Select machine learning classifiers will be deployed to generate models for the prediction of SAMS and the best-performing model will be selected based on model performance. RESULTS/ANTICIPATED RESULTS: The expected outcomes include generation of a fine-tuned NLP algorithm that can rigorously identify SAMS occurrences within EHRs. Further, we anticipate having a practical risk model that accurately predicts patients’risks of developing SAMS when taking statins. DISCUSSION/SIGNIFICANCE: The positive and translational impact of our research will be to equip healthcare providers with such informatics tools to improve statin adherence, ultimately promoting patient optimal health and outcomes by maximizing the tolerance and thus realizing the therapeutic benefits of statins.

OBJECTIVES/GOALS: The objective of the study was to determine whether CLP altered genes associated with mitochondrial function in the diaphragm. METHODS/STUDY POPULATION: A rodent cecal-ligation and puncture (CLP) model used to mimic sepsis-induced critical illness. The CLP model involved ligation of 50% of the cecum below the ileocecal valve in adult C57BL6 mice, followed by needle puncture of the cecum resulting in mid-grade sepsis. Mice survived for 48 hours or more, following injury. Diaphragm and limb muscles were harvested 24 hours following CLP (N = 6) and following a sham CLP procedure (N = 6). RESULTS/ANTICIPATED RESULTS: Gene expression of mitochondrial related genes (mef2c, myh1, pgc1-α), were significantly decreased in the diaphragm of CLP injured animals when compared to controls. In addition, ubiquitin ligases, genes associated with skeletal muscle atrophy murf1 and atrogin were increased in the diaphragm 24 hours after injury (p< 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: Our results indicate that sepsis-induced critical illness significantly impacts the expression of genes implicated in mitochondrial homeostasis and atrophy. Ongoing studies will identify whether CLP injury decreases skeletal muscle mitochondrial function.

OBJECTIVES/GOALS: MGMT methylation status is used to predict the response to TMZ. However, a subpopulation of patients lacking MGMT methylation still respond to TMZ. We applied omics approaches and functional studies to a cohort of GBM patients to identify novel markers that may more accurately predict TMZ response. METHODS/STUDY POPULATION: We applied a combination of omics approaches and functional studies to a cohort of GBM patients to search for novel markers that would predict the response to TMZ treatment more accurately than traditional markers. Using a set of 47 primary and secondary GBM tumor samples, we employed comparative transcriptomics, whole exome sequencing, data independent acquisition (DIA) proteomics, and phosphoproteomics to look for DNA mutations and changes in gene expression and/or protein expression that correlated to response or non-response to TMZ. Subsequently, we performed functional studies and analyzed patient treatment data to validate our results. RESULTS/ANTICIPATED RESULTS: This study is in early stage, but we anticipate that our combination of methods may allow us to identify and validate at least one novel biomarker for TMZ response in patient GBM. For example, comparative transcriptomics or phosphoproteomics may identify a previously unrecognized gene or protein over/under-expression in a subset of patients. In this case we will validate findings using western blotting, IHC staining, and through siRNA of target gene/protein on patient derived GBM cells to examine if removal of this marker leads to TMZ sensitivity. We will further confirm prediction of marker correlation through comparison with matched patient treatment data. DISCUSSION/SIGNIFICANCE: The identification of improved biomarkers to predict response to TMZ treatment is a discovery that could rapidly become standard of care for GBM patients. It would ensure that all responders receive TMZ and avoid exposing nonresponders unnecessarily to TMZ and its potential side effects.

OBJECTIVES/GOALS: We previously reported that genetic polymorphisms in the beta-catenin gene (CTNNB) are associated with the development of Clostridiodes difficile colitis during autologous stem cell transplantation (https://www-ncbi-nlm-nih-gov.proxy.libraries.uc.edu/pubmed/29594489). To biological validate these findings, we sought to evaluate the development of chemotherapy-associated Clostridiodes difficile infections by assessing the effect of C.difficile toxin B (TcdB) and of using melphalan in beta-catenin protein expression in Caco2 cells. METHODS/STUDY POPULATION: To determine the effect of melphalan and/or C.difficile toxin B on expression of Beta-catenin from human gut epithelial cells:

• Adenocarcinoma cells (Caco-2) cells were seeded and allowed to grow into monolayers

• Monolayers were treated with PBS, TcdB, melphalan and/or TcdB + melphalan for 24 hours and then washed with PBS

• Immunofluorescence was measured on the monolayers to visualize three markers -DAPI-Nuclear Stain (blue),Actin-ccytoskeletal stain (red), B-Catenin (green)

• Analysis of images with ImageJ (NIH). Statistical analysis of the effect of TcdB and/or melphalan on β-catenin protein levels was determined by One-way ANOVA

OBJECTIVES/SPECIFIC AIMS: This study aims to describe adaptability in methods used to apply community input to programming within the field of translational science. The outcomes of community informed programming include opportunities for innovative projects and approaches, and better responsiveness to community needs. It is anticipated that this will result in greater community involvement in research, moving towards greater health equity. METHODS/STUDY POPULATION: The SC CTSI is situated in urban Los Angeles, one of the most diverse communities in the world. Eight SC CTSI Community Engagement Core initiatives that employ community partnership are illustrated. The activities include social marketing campaigns for cervical cancer prevention; use of community-embedded research ambassadors to increase scientific literacy in Latino and Black/African-American communities; use of innovative technologies to educate pediatric patients and families about clinical research; working with the entertainment industry to promote clinical research in popular television shows; a community advisory board that is tailored and embedded in each CSTA core group; a community based research dissemination program; an ad-hoc community advisory group assembled to adapt a research 101 curriculum for Black/African-American communities; and a series of listening sessions conducted throughout Los Angeles. RESULTS/ANTICIPATED RESULTS: Integration of community voices provide direction for future planning, programming and execution of all referenced initiatives. Ultimately, the goal for these discussions with community members is to develop innovative approaches to CTSA programming. DISCUSSION/SIGNIFICANCE OF IMPACT: Racial and ethnic minorities continue to experience underrepresentation in clinical research trials. CTSAs have been tasked with addressing barriers that have historically led to disparities in research participation, and by extension, the effectiveness of medical interventions in diverse populations. Community input is an invaluable source for knowledge and innovative ideas in how to increase involvement in various aspects of the research process, including dissemination, recruitment and enrollment in clinical trials. CTSAs have increasingly augmented Community Engagement programs within their respective cores to address population disparities. The approaches used to engage communities require an element of fluidity and flexibility, and a reliance on the input of community members, in order to maintain relevant and desired community engagement practices.

OBJECTIVES/GOALS: To maximize health outcomes from their work, basic researchers must understand the process by which lab discoveries are translated into clinical care. We developed an academic course designed to provide students in our Clinical and Translational Sciences PhD program with an in-depth understanding of translational applications of basic research. METHODS/STUDY POPULATION: A preliminary needs assessment was done with students, educators, and clinicians to identify the course content. Based on these data, didactic modules including research question identification, research team development, participant recruitment, and research data collection were piloted in a synchronous, virtual course. Then, for 6 weeks, students shadowed clinical mentors who worked in the students research areas. Finally, with their mentors, students developed and presented clinical research protocols. Student pre- and post-course surveys gauged alignment of course objectives and learning outcomes. A post-course, focus group with students gathered feedback on course content, structure, and students confidence in implementing their experiences from the course into real-world settings. RESULTS/ANTICIPATED RESULTS: Six MD/PhD and PhD students participated in the pilot course. Pre/post-assessments (n=4) showed students were more confident in clinical question/research protocol formulation, development of patient recruitment/enrollment strategies, and integration of research methodologies into their research projects after completing the course. Students asked for additional content on budgeting and grant funding. Post-course focus group participants (n=2) appreciated the experience of writing a clinical protocol and the flipped classroom teaching style, which allowed them to network with clinical faculty leading didactic sessions. Students also noted course content was relevant and motivating, although they suggested adding content about clinical trials measures to enhance their shadowing experiences. DISCUSSION/SIGNIFICANCE: A course that combines didactic and clinical experiential training provides a robust, translational research foundation for basic scientists. This training is critical to help them contribute to the effective/efficient translation of lab discoveries to clinical practice. Future course development will include students from other PhD programs.

OBJECTIVES/GOALS: To investigate digital behavior patterns before cerebrovascular disease (CeVD), we compared accelerometer-measured physical activity (PA) phenotypes of future CeVD patients versus controls in UK Biobank. METHODS/STUDY POPULATION: Accelerometer data from 76,525 eligible participants monitored for 7-days (Jan. 2013-Dec. 2015) was classified into four categories: sedentary, light PA (LPA), moderate-to-vigorous PA (MVPA), and sleep. Covariables and diagnoses were defined using baseline data and patient records. Daily PA patterns associated with incident CeVD were compared to controls using negative binomial regression models. RESULTS/ANTICIPATED RESULTS: Adult participants with future CeVD (n = 2,163) spent 4.4% less time in MVPA (Incident Rate-Ratio (IRR) 0.956; 95% CI = 0.923-0.992; p = 0.016) compared to controls. During 0:00-5:59h (midnight to 5:59AM), future CeVD patients were less likely to sleep (IRR = 0.985; 95% CI = 0.977-0.992; p <0.001) but more likely to be sedentary (IRR = 1.189; 95% CI = 1.098-1.290; p <0.001) or in LPA (IRR = 1.108; 95% CI = 1.015-1.211; p <0.001). In subgroup analyses, decreased MVPA was observed in current/former smokers (IRR = 0.887; 95% CI = 0.819-0.963), males (IRR = 0.931; 95% CI = 0.870-0.997), and the unemployed/retired (IRR = 0.923; 95% CI = 0.856-0.998), an effect more pronounced in depressed patients (p for interaction = 0.044) and prolonged (> 2 hr/day) screen users (p for interaction = 0.018). DISCUSSION/SIGNIFICANCE: The digital phenotype of PA prior to CeVD is characterized by less sleep during 0:00-5:59h and less daily MVPA, demonstrating the utility of accelerometer data in identifying candidates for preventative intervention.

Objectives/Goals: The prevalence of type 2 diabetes (T2D) in adolescents is rising, presenting unique challenges for recruitment in clinical research – particularly among adolescents who belong to minoritized race/ethnic backgrounds. The primary aims of our study are to identify and address barriers to recruiting adolescents, particularly around community trust. Methods/Study Population: This study employed a two-phase approach to evaluate recruitment strategies for underrepresented adolescent populations in genomics studies of T2D. After IRB approval, in Phase 1, we utilized the electronic medical record (EMR) system at Children’s of Alabama to prescreen based on inclusion criteria (adolescents aged 12–18 years with T2D). Recruitment efforts were tailored to address barriers unique to underrepresented populations, such as flexibility in scheduling contact times to accommodate family availability. In Phase 2, we are implementing and assessing the effectiveness of a peer recruitment model, wherein adolescents from underrepresented groups are trained to engage peers within their community. Focus groups (6–8 participants per group) will explore facilitators and barriers to recruitment. Results/Anticipated Results: Phase 1 revealed that EMR prescreening effectively identified eligible participants, and direct outreach (phone calls, face-to-face contact during clinical visits) significantly improved recruitment success, especially among underrepresented adolescents. Flexible scheduling and consolidating study appointments enhanced participation, addressing logistical challenges like geographic distance. However, frequent changes in contact information (phone and email information) created barriers. In Phase 2, early focus group results suggest that peer recruitment is promising, with trust and community engagement being key factors. Adolescents recruited by peers were more likely to participate, though logistical hurdles such as transportation and family concerns remain. Discussion/Significance of Impact: Our study demonstrates the value of personalized outreach and prescreening in improving recruitment among adolescents. These strategies can engage communities that include Black, Indigenous, and People of Colored. These findings underscore the need for flexible recruitment strategies to ensure their participation in T2D adolescent research.

ABSTRACT IMPACT: Screening the effect of thousands of non-coding genetic variants will help identify variants important in the etiology of diseases OBJECTIVES/GOALS: Massively parallel reporter assays (MPRAs) can experimentally evaluate the impact of genetic variants on gene expression. In this study, our objective was to systematically evaluate the functional activity of 3’-UTR SNPs associated with neurological disorders and use those results to help understand their contributions to disease etiology. METHODS/STUDY POPULATION: To choose variants to evaluate with the MPRA, we first gathered SNPs from the GWAS Catalog that were associated with any neurological disorder trait with p-value < 10-5. For each SNP, we identified the region that was in linkage disequilibrium (r2 > 0.8) and retrieved all the common 3’-UTR SNPs (allele-frequency > 0.05) within that region. We used an MPRA to measure the impact of these 3’-UTR variants in SH-SY5Y neuroblastoma cells and a microglial cell line. These results were then used to train a deep-learning model to predict the impact of variants and identify features that contribute to the predictions. RESULTS/ANTICIPATED RESULTS: Of the 13,515 3’-UTR SNPs tested, 400 and 657 significantly impacted gene expression in SH-SY5Y and microglia, respectively. Of the 84 SNPs significantly impacted in both cells, the direction of impact was the same in 81. The direction of eQTL in GTEx tissues agreed with the assay SNP effect in SH-SY5Y cells but not microglial cells. The deep-learning model predicted sequence activity level correlated with the experimental activity level (Spearman’s corr = 0.45). The deep-learning model identified several predictive motifs similar to motifs of RNA-binding proteins. DISCUSSION/SIGNIFICANCE OF FINDINGS: This study demonstrates that MPRAs can be used to evaluate the effect of non-coding variants, and the results can be used to train a machine learning model and interpret its predictions. Together, these can help identify causal variants and further understand the etiology of diseases.

OBJECTIVES/GOALS: Despite calls for the integration of Education for Sustainable Healthcare (ESH) into health professional training programs, most curricula have yet to adapt accordingly. This qualitative study sought to understand faculty perspectives on ESH knowledge, interest, barriers, and facilitators. METHODS/STUDY POPULATION: From 2018 to 2020, 71 health professional education faculty from 6 University of California (UC) campuses participated in ESH integration workshops. Using purposeful sampling based on gender and campus, a subset of workshop faculty participants were selected to participate in individual interviews. Interviews were conducted via Zoom using a structured interview guide, eliciting participants experience integrating ESH, perceived barriers and facilitators, and perspectives on student, faculty, and health science leadership knowledge and interest. Transcripts were double coded with an inductive-deductive approach using Dedoose, reconciled, and analyzed to identify themes. RESULTS/ANTICIPATED RESULTS: Participants included 17 faculty at 6 UC campuses representing diverse health disciplines. Although participants noted high general awareness of and interest in climate change among students and faculty, they observed a lack of specific, health-relevant knowledge, resulting in discomfort communicating with others on climate and health. Perceived barriers to expansion of ESH included limited curricular space, competing topics, and lack of faculty expertise. Participants posited that framing climate change in health terms, establishing learning objectives and protected faculty time, identifying connections to ESH within existing research and curriculum, and obtaining commitments from campus leadership would facilitate successful ESH integration. DISCUSSION/SIGNIFICANCE: Our findings reinforce student and faculty interest in ESH curricular integration and identify important barriers and facilitators, lending context for educators planning ESH infusion. Training of faculty on climate health is urgently needed for ESH integration in health professional education.

OBJECTIVES/GOALS: Our goal was to distinguish SARS-CoV-2 CD8+ T cell epitopes of spike, membrane, and nucleocapsid products in 27 of the most frequent HLA-Aand -B alleles.We hypothesize that differences mediated by variation in SARS-CoV-2 and host HLA genetics affect the differential clinical severity and presentation of acute infection and PASC. METHODS/STUDY POPULATION: Genomic sequences of SARS-CoV-2 variants were blasted against the original Wuhan strain using Ensembl’s SARS-CoV-2 browser. We examined 16 COVID variants: 2 Alpha (B.1 and B.1.1.7), 5 Delta (AY.100, AY.25, AY.3, AY.3.1, and AY.44), and 9 Omicron (BA.1, BA.1.1, BA.2, BA.4, BA.5, BQ.1, BQ.1.1, XBB.1, and XBB.1.5), sequenced from the Louisiana patient population. cDNA sequences were translated using the Expasy tool. To predict MHC-I epitope binding, we used the Immune Epitope Database and Analysis Resource, via TepiTool utilizing the IEDB recommended default prediction and the 27 most frequent HLA-A and -B alleles. In silico peptide docking was conducted on FoldX, utilizing HLA-B*15:01 structures (n= 7) from the Protein Data Bank. RESULTS/ANTICIPATED RESULTS: CD8+ epitope conservation was estimated at 87.6-96.5% in S, 92.5-99.6% in M, and 94.6-99% for N. As the virus mutated, an increasing proportion of S epitopes experienced reduced predicted binding affinity: 70% of Omicron BQ.1- XBB.1.5 S epitopes experienced decreased predicted binding, as compared to ~ 3% and ~15% in Delta AY.100-AY.44 and Omicron BA.1-BA.5 respectively. Additionally, we identified several novel candidate haplotypes that may be susceptible to severe disease, notably HLA-A*32:01, -A*26:01, -B*58:01, and -B*53:01, and relatively protected from disease, such as -A*01:01, -A*02:01,-A*31:01, -B*15:01, -B*40:01, -B*44:03, and -B*57:01. In silico analysis of COVID peptides and HLA-B*15:01, a common allotype in the United States, largely matched predicted binding patterns. DISCUSSION/SIGNIFICANCE: To elicit long term COVID-19 immunity and prevent PASC, it is important to understand the relationship between T-cells, viral variants, and HLA genetics. This project is one of the first to explore the interaction between CD8+ epitope diversity and viral genetics for the majority of the United States population.