OBJECTIVES/GOALS: To evaluate the clarity of plain-language informed consent forms (ICF) created using a browser-based tool called the ICF Navigator, we solicited feedback from two community review boards (CRB) to ensure the resulting ICF met the informational needs of all potential participants, including those with limited health literacy skills. METHODS/STUDY POPULATION: Community-engaged research highlights the importance of involving community members in the planning and execution of translational research projects. Virtual discussions were held to elicit feedback from two separate CRBs on the understandability of an ICF that was generated using an online, browser-based tool that we designed to aid researchers in the creation of plain-language ICFs. CRBs included representation of diverse communities from across the state of Arkansas, including individuals who may have limited health literacy skills, those with and without prior experience participating in clinical research projects, members living in rural and urban settings, and those whose race or ethnicity have been traditionally underrepresented among clinical research participants. RESULTS/ANTICIPATED RESULTS: CRB feedback was used to inform actionable improvements to the tool, such as removing content redundancies and embedding tips to guide researchers on how best to optimize the clarity and understandability of resulting ICFs. Program refinements in response to the feedback have been implemented and will be evaluated in another round of CRB discussions in early 2023. Feedback from this follow-up CRB session will also be presented in addition to a discussion of how the feedback was used to improve the online tool, which will ultimately be available for free use by other institutions. DISCUSSION/SIGNIFICANCE: The use of community feedback to optimize the functionality of the ICF Navigator demonstrates the value of CRBs for ensuring that ICFs are culturally salient and readily understandable by all potential research participants, particularly those who may have limited health literacy skills, thereby promoting more equitable opportunities for all.

OBJECTIVES/GOALS: Team science is a focus of the University of Florida Clinical and Translational Science Institute (UF CTSI) Learning Health System Initiative. Citizen Scientists (CSs) are integral research partners who provide pragmatic feedback. The UF Health Cancer Center (UFHCC) aspired to adopt a similar approach to research, starting with onboarding CSs. METHODS/STUDY POPULATION: To understand the content with which they would be engaging through the CTSI, CSs first became certified through a self-paced online curriculum focused on clinical research basics. UFHCC envisioned their onboarding as a companion piece to this first course, and CSs must complete both courses to work in cancer-focused research. The new CS Cancer Curriculum consists of a mix of didactic lessons with quizzes, case studies, a behind-the-scenes look at a research lab meeting, and interviews with CSs. As with the clinical research course, the cancer course was co-developed alongside the CSs and utilized the ADDIE (Analysis, Design, Development, Implementation, Evaluation) instructional design model. The course was implemented with UF CSs from July to September 2021 through Canvas. RESULTS/ANTICIPATED RESULTS: For the nine CSs completing this pilot test, scores for all didactic quizzes across the course were mostly high. Two CSs scored a perfect 100%, three missed only one question (98%), and two CSs missed two questions (96%). A course evaluation was completed by eight of the CSs and determined that most (86%) felt that they were capable of applying what they had learned. An additional 75% felt the course empowered them to advocate for the needs of all stakeholders involved in cancer research. Qualitative responses on the evaluation found that the course helped CSs better relate to the challenges faced by other stakeholders (patients, clinicians, caregivers) and helped them conceptualize how they could contribute to cancer research. DISCUSSION/SIGNIFICANCE: The partnerships within the UF CTSI have a direct impact on patient care through research studies in Florida and nationwide. CSs can be overlooked by researchers unfamiliar with concepts of a learning health system, including those in cancer research. By engaging these stakeholders, we may soon see similar impacts to cancer-related patient care.

OBJECTIVES/GOALS: Patients on chronic opioids face gaps in transitions of care in the time following surgery, increasing the risk for adverse events, specifically high-risk opioid prescribing. The objective of this study is to determine how rates of high-risk prescribing differ between patients with public and private insurance. METHODS/STUDY POPULATION: A retrospective cohort study of 1,435 adult patients with preoperative chronic opioid use on Medicaid or commercial insurance who underwent surgery between November 2017 and February 2021. Patients were identified using the Michigan Surgical Quality Collaborative (MSQC) database, a collection of perioperative data from 70 hospitals across the state of Michigan. Data from the MSQC were merged with Michigan’s prescription drug monitoring program to provide additional information on pre- and postoperative opioid prescribing. Multivariable logistic regression was used to assess high-risk prescribing by the presence of a preoperative usual prescriber and insurance type. RESULTS/ANTICIPATED RESULTS: Overall, 22.7% of patients on private insurance and 23.6% of patients on Medicaid fulfilled criteria for new, postoperative high-risk prescribing. Among criteria for high-risk prescribing, multiple prescribers was the most significant contributor (private insurance: 17.4%, Medicaid: 18.9%). Patients on Medicaid insurance did not have increased odds of new postoperative high-risk prescribing (OR = 1.067, 95% CI: 0.813-1.402). While fewer patients on Medicaid had a preoperative usual prescriber (86.9% and 90.9% respectively, p = 0.015), there was no significant difference between the two insurance types in baseline rates of high-risk prescribing prior to surgery (private insurance: 43.4%, Medicaid: 46.0%, p = 0.352). DISCUSSION/SIGNIFICANCE: While we do not observe disparities in high-risk prescribing between insurance types, rates of high-risk prescribing postoperatively are high across payer types. Further studies to determine the factors driving rates of high-risk opioid prescribing among patients with chronic opioid use are needed to identify areas for future intervention.

OBJECTIVES/GOALS: African-Americans are at increased risk for nutrition disparities; home cooking is a strategy to optimize dietary quality. to develop a cooking intervention, a mixed-methods community-based participatory, acceptability study was conducted to understand cooking behaviors, options for intervention content, and implementation factors. METHODS/STUDY POPULATION: Self-identified African-American adults were recruited from a larger community-based study within Washington, D.C. Five moderated virtual focus groups with four participants in each group were conducted in March and April of 2021. A semi-structured moderator’s guide focused on cooking, meal habits, food choices, and the proposed cooking intervention was utilized. Qualitative data collected were verbatim transcriptions and notes from research team members. Thematic analysis was conducted using an iterative process among research team members. Participant validation interviews were conducted following the research team analysis. Electronic self-administered surveys were used to measure demographic, food environment, cooking behavior, health behavior, and psychosocial variables. RESULTS/ANTICIPATED RESULTS: Study participants (n=20 females, mean age 60.1 years) lived in low-food resource neighborhoods but reported high food security (n=14). Barriers to the intervention included traveling distances to other neighborhoods for produce, poor quality of vegetables and fruit within neighborhood stores, lack of trustworthiness from neighborhood store experiences, perception of decreased cooking skills, and competing priorities related to time and weekday schedules. Motivators included health promotion for self, family, community, and enjoyment from cooking. Virtual or in-person sessions were suggested. Intervention options included costs of recipe ingredients, using recipes with ingredient flexibility, nutrition information, and provisioning of or compensation for ingredients used in virtual classes. DISCUSSION/SIGNIFICANCE: Use of virtual focus groups for a participant-based design of a cooking intervention among African American adults living in low-food access neighborhoods provided acceptability results that were food and home environments contextual and provided barriers and motivators to participation and implementation of behavior from the intervention.

OBJECTIVES/GOALS: Alabama has the 3rd highest maternal mortality ratio in the U.S., with more than 50% of deaths occurring postpartum. There is little evidence on the prevalence or equity of postpartum care use in Alabama. This mixed methods study examines relationships between patient and provider factors and access to, use of, and racial disparities in postpartum care. METHODS/STUDY POPULATION: I will use a sequential explanatory mixed methods design. In the quantitative phase I will analyze an integrated electronic health record and human resource dataset to identify patient and provider factors that have a relationship with receipt of at least one postpartum visit within 12 weeks of delivery in a cross-sectional, retrospective cohort of 30,000 obstetric patients in Alabama. In the qualitative phase I will describe the postpartum experiences of obstetric patients who identify as Black or African American who received or did not receive at least one postpartum visit within 12 weeks of childbirth. In the integration phase I will draw synthesized conclusions about how the results of both phases describe predictors of and barriers and facilitators to postpartum care for Black birthing people in Alabama. RESULTS/ANTICIPATED RESULTS: I will identify relationships between patient factors (e.g., race, racial concordance with primary provider, insurance status, age, parity, type of delivery, Area Deprivation Index, presence of a chronic condition or severe morbidity) and patient receipt of postpartum care. I will also explore whether health care provider factors (e.g., race, racial concordance with the patient, age, gender, provider type, years of experience) predict patient receipt of postpartum care in this retrospective cohort. In the qualitative phase, I will explore the experiences and perceptions of birthing people who identify as Black or African-American that help explain the relationships between patient and provider factors and receipt of postpartum care identified in the quantitative phase. DISCUSSION/SIGNIFICANCE: More than 50% of maternal death occurs after childbirth. Postpartum care is critical to birthing people’s survival, especially in states with high maternal mortality. This study will fill a gap in knowledge about factors that have a relationship with equitable postpartum care in Alabama.

OBJECTIVES/GOALS: We have developed a community engagement model that embraces several core values: bi-directional, co-equal, co-created, inclusive, culturally centered, and trust. Our promotoras de salud and community health workers (CHWs) facilitate listening sessions to understand the most pressing health needs and concerns as perceived by the community. METHODS/STUDY POPULATION: Our Southern California CTSI Community Engagement team includes promotoras de salud and community health workers who are trusted individuals within our targeted communities–Latino and Black populations in South and the Eastside of Los Angeles. Listening sessions identify the community’s perceived health needs and concerns, and our team in turn delivers a workshop series that addresses those concerns. Workshops are co-developed by community members, offered in English and Spanish, and delivered both virtually and in-person in community venues. Workshop topics have included mental health, depression, anxiety in children; child development and autism; COVID-19 testing, variants, and vaccines; monkeypox; and a primer on understanding clinical research. RESULTS/ANTICIPATED RESULTS: In 2022, we held 49 workshops that delivered evidence-based information including how to access needed resources. We had a total of 1212 participants, 60% of whom were returning, meaning they had attended at least one prior workshop. An evaluation of the workshop program demonstrated that our promotoras de salud and community health workers are trusted by the community and that workshops are meeting community needs. 677 surveys were completed by participants, 87.5% of whom were female. 85% of respondents stated that they will use the information they learned in their daily life, and 90% said they would attend another workshop. Feedback from the evaluation will inform future workshop format and topics. DISCUSSION/SIGNIFICANCE: Academic-community partnerships are key to advancing health equity, especially in diverse and under-resourced communities. This is one model of community engagement, which includes promotoras de salud and CHWs that was designed to build trust, empower individuals through education, and to address the health needs as defined by the community.

OBJECTIVES/GOALS: We seek to describe the challenges to mental healthcare access and utilization among Somali refugees in Minnesota. The objectives of this study are 1) to characterize beliefs and attitudes about mental illness and 2) identify barriers, either personally experienced or perceived, to utilizing mental health services among Somali refugees. METHODS/STUDY POPULATION: Mental health challenges are of particular concern among Somali refugees, who have been found to have rates of PTSD as high as 50%. However, Somali refugees are reported to underutilize mental healthcare. We will recruit 20-25 Somali refugee women and men, who are 18 years or older and reside in the Twin Cities, to participate in one on one interviews. Participants will be asked about their perception of barriers to mental healthcare services, and their beliefs about mental illness and treatment. We will transcribe the interviews, code them, and identify key themes. A community advisory board will be directly involved in the research design, recruitment, interview instrument development, interpretation of findings, and dissemination of project materials as part of our community engagement protocol. RESULTS/ANTICIPATED RESULTS: When completed we expect to identify mental health beliefs and barriers to mental healthcare utilization. The long-term goal of this work is to reduce the substantial mental health morbidity among Somali refugees. Common barriers to research participation that we expect are mistrust, financial constraints, fear of unintended outcomes, stigma about participating in research, and fear of deportation or concern of immigration status. We plan to address these barriers by hiring bilingual Somali recruiter/interviewer, translating study materials, reassuring confidentiality of participant’s information, providing a $50 incentive, and implementing community advisory board’s input on study design and recruitment sites. DISCUSSION/SIGNIFICANCE: Translational Impact: Findings from interviews will be disseminated and evaluated by members of the community and providers. Recommendations based on our findings can be applied in mental healthcare practice to reduce identified barriers. Community dissemination can also promote the destigmatization of mental healthcare in the Somali community.

OBJECTIVES/GOALS: All IN for Health’s Advisory Board is an active board providing advice on strategic directions, feedback, contributing ideas, and accountability. Most recently, the HAB had 4 openings. A call to our community of over 13,000 individuals was published. We received 488 applications. We share the lessons in motivations and interests shared respondents. METHODS/STUDY POPULATION: Four hundred eighty-eight individuals submitted applications for participation in the All IN for Health Advisory Board. The call went out to the All IN for Health volunteer research network comprised of community members across the state.The call mentioned a nominal compensation of $50/hour, and time commitment of a 2-hour board meetings via video conferencing every 3 months. The application process included the following questions: 1. What lived experiences and/or personal interests have motivated you to be involved in All IN for Health? 2. Please explain why you are interested in being an All IN for Health Advisory Board member? RESULTS/ANTICIPATED RESULTS: We organize the findings in two categories: Motivation and Interests. The motivation category refers to individual’s motivation to be part of the All IN for Health initiative. Interest category referred to individual’s specific interest in participating in the All IN for Health Advisory Board. Individuals were motivated to participate based on (1) family or friend diagnosis, (2) personal diagnosis, (3) roles as caregivers, (4) desire to impact change and advocacy, (5) role as health professional, and (6) previous participation in research. Interests followed similar themes with additional categories of sharing their experience, previous research and board experience, as well as a desire to educate future generations of researchers. DISCUSSION/SIGNIFICANCE: We share the narratives honoring individual stories for the top three motivations and interest. This information can be used for recruitment to boards, research participation, and healthcare advocacy, and highlights importance of sharing successes and challenges to the processes of forming and sustaining effective boards.

OBJECTIVES/GOALS: The critical role that trust plays continues to be documented and highlights the gap in understanding the extent to which initiatives such as All IN for Health from the Indiana Clinical and Translational Institute (I-CTSI) can improve trust between the community and researchers. We discuss measures of trust and evaluate how we may improve recruitment. METHODS/STUDY POPULATION: In this study, we invited over 13000 volunteers from the All IN for Health research participant registry. Six hundred and sixty-three (663) respondents participated in the survey. The Relationship of Trust and Research Engagement Survey included three validated surveys: Distrust in Healthcare Organizations, the Trust in Medical Researchers scale by Hall et. al, and Patient Trust in Medical Researchers by Mainous et al.. The 36-item survey also included open-text questions. RESULTS/ANTICIPATED RESULTS: Based on preliminary results 74% agreed it’s safe to be in medical research, yet 79% had never been asked to participate in medical research by their doctor. Sixty percent believed that HCOs put money above patients’ needs. Forty percent agreed that doctors do medical research for selfish reasons. Fifty percent disagree that patients get the same medical treatment regardless of race/ethnicity. Moreover, 28 % agree that medical researchers act differently toward minorities. Between 9 and 11% believe that researchers select minorities for their most dangerous studies and some projects are secretly designed to expose minoritized groups to diseases. Our next step is to disaggregate the data by race and ethnicity and evaluate these answers. DISCUSSION/SIGNIFICANCE: This study's population willingly engaged in a research registry making their diminished trust quite alarming. Amongst the general population, trust in scientists is now below pre-pandemic levels. We must critically assess our own trustworthiness, and critically reflect on the authenticity of our efforts.

OBJECTIVES/GOALS: COVID-19 disproportionately impacts rural communities of color. Socioeconomic status, occupation and chronic illnesses lead to worse COVID-19 outcomes. This study identifies motivators and barriers of COVID-19 vaccine uptake in the Latino and Flathead Reservation agricultural communities. METHODS/STUDY POPULATION: Thirty key informant interviews and 6 focus groups (N=39 focus group participants) were conducted with community and tribal leaders using an interview guide informed by the Theory of Planned Behavior, Social Cognitive Theory, and the Social Contextual Factor Frameworks. The interview guide was designed to understand the motivators and barriers of COVID-19 vaccine uptake. The Community Advisory Board, community investigators and community health workers from the community reviewed and revised the guide. A codebook applied deductive coding to informant responses, followed by an inductive, constant comparison approach. Three analysts met to refine the codebook and conduct inter-rater agreement. RESULTS/ANTICIPATED RESULTS: Participants from Flathead reservations and Yakima frequently noted a desire to protect one’s self, family and elders. This significant motivator encouraged individuals to receive the COVID-19 vaccine, despite sincere vaccine concerns and government rollout. Barriers included concerns regarding rumored, serious or rare side effects, speed of vaccine development and misinformation. Key differences exist between both communities. Yakima participants noted religious concerns and ID requirements as major barriers. Flathead reservation participants noted distrust and historical trauma of the U.S. government and issues with access (e.g. transportation, technology). DISCUSSION/SIGNIFICANCE: The pandemic disproportionately impacts vulnerable communities in agricultural settings. Participants in both communities felt vaccine availability had outpaced uptake. Clearly, culturally sensitive education and respectful communication would be key in addressing vaccine concerns and improving vaccine uptake.

OBJECTIVES/GOALS: To investigate nursing homes (NHs) acquired by private equity (PE) firms and estimate the effect of PE NH acquisitions on NH care quality for NH residents and whether PE NH acquisitions exacerbate racial inequities in quality of care. METHODS/STUDY POPULATION: My research estimates the causal effect of NH PE acquisitions on NH access and quality of care for NH residents and whether acquisitions exacerbate racial disparities in NH care for about 115 PE-owned NHs in the US, measuring the quality of care at the facility survey year level, and using an array of NH-level data sources. I identified 115 PE-owned NHs (treatment group) and 665 non-PE-owned NHs between 2003 and 2010, using the Online Survey Certification and Reporting database to obtain facility characteristics. I compare facility characteristics (e.g., payer mix, staffing levels, and quality measures such as pressure ulcers, unexpected weight loss, acuity, and health deficiencies). I will then test whether effects differ by race, with hypotheses informed by Public Health Critical Race Praxis approach. RESULTS/ANTICIPATED RESULTS: Preliminary results show that staffing levels differ between PE and non-PE-owned NHs in a way that aligns with a shift in focus toward the Medicare population i.e. short stays. We also find that deficiencies increased in PE-owned NHs compared to non-PE-owned NHs. We expect that PE acquisitions may lead to slightly widening racial disparities in NH care quality. Results may show that PE-owned NHs have a higher share of low-rated, high-BIPOC facilities. In weak markets, PE-owned NHs may have a higher share of BIPOC residents compared to highly competitive markets. This is because PE managers may prioritize cost over quality by cutting services. However, since quality measures are self-reported, except for measures related to deficiencies, this predicted lower quality of care may not be evident in observed data. DISCUSSION/SIGNIFICANCE: Understanding how PE ownership impacts nursing home care quality for long-stay residents, especially those funded by Medicaid, can help develop intervention strategies to effectively mitigate racial inequities in NH care, as Medicaid funded NH residents are more likely to be Black, Indigenous, and people of color.

OBJECTIVES/GOALS: Our objective is to assess whether street race is a stronger predictor of sleep quality and sleep hours than self-reported race. We also seek to understand whether the association between race and sleep quality/hours can be explained by experiences of microaggressions. METHODS/STUDY POPULATION: This study uses data from the National Couples’ Health and Time Study (NCHAT), a population-representative sample of 20–60 year-olds (N=3,642) who were married or cohabiting during 2020-2021 when the COVID-19 pandemic disproportionately negatively impacted racial and ethnic minorities in the U.S. (Boserup et al., Yip et al.). During this time, incidents of racial trauma increased (Tessler et al.). Using NCHAT data we examine whether street race is a stronger predictor of sleep quality and sleep hours than self-reported race. We also seek to understand whether the association between race and sleep quality/hours can be explained by experiences of microaggressions. RESULTS/ANTICIPATED RESULTS: Results show that microaggressions mediate the link between identifying as Black and being perceived as Black or Asian and sleep quality/hours. Identifying as Black and being perceived as Black or Asian, compared with non-Latinx White respondents, is associated with more frequent microaggressions. More microaggressions are associated with poorer sleep quality and fewer sleep hours. Asian street race is a marginally better predictor of microaggressions than self-reported race. In all models, microaggressions are associated with poorer sleep quality and less sleep hours. DISCUSSION/SIGNIFICANCE: With a growing non-white population, the wellbeing of our future generations is in everyone’s best interest. Poor sleep increases the risk of cardiovascular disease, diabetes, obesity, and cancer. The United States spends $93 billion in excess medical care costs due to health disparities.

OBJECTIVES/GOALS: 1. Describe method of Community Based System Dynamics. 2. Describe CBSD as used in addressing structural racism in a previously redlined community. 3. Review example causal loop diagrams and simulation models developed using CBSD focused on structural racism as a social determinant of health. METHODS/STUDY POPULATION: Community Based System Dynamics (group model building and computer simulation development). Cuyahoga County, Ohio, a previously redlined community of more than 1.2 million people. RESULTS/ANTICIPATED RESULTS:Actionable Community identified leverage points for action to mitigate structural racism. Computer simulation models built on causal loop diagrams built by Community members with loved experience. DISCUSSION/SIGNIFICANCE: Structural and social determinants of health, such as racism, have profound impacts on the health of individuals and populations, however they remain challenging to address in pragmatic ways. CBSD is a novel method to engage community members not proximal to the impact of structural racism in generating maps of the complex, dynamic system they live in.

Precision Medicine/Health

OBJECTIVES/GOALS: Overall response rates for metastatic uveal melanoma (UM), regardless of treatment modality, are less than 20%, highlighting an urgent need for novel therapies. Herein, we present a UM patient-derived organoid (PDO) biobank as a novel platform for translational research. METHODS/STUDY POPULATION: Patients with primary choroidal or ciliochoroidal UM undergoing enucleation from 7/1/2019-9/30/2022 were invited to enroll. Tumor tissue was harvested within 30 minutes of globe removal. Cells were isolated using the human tumor isolation kit and gentleMACS dissociation protocol (Miltenyi Biotech). PDOs were placed on Cultrex-coated multiwell plates and cultured in supplemented RPMI media. DNA and RNA were isolated using kits from Zymo Research. Exon-enriched libraries and RNA were sequenced using an Illumina HiSeq 4000. Immunohistochemistry (IHC) assessed the following histone post-translational modifications: H3K4me1/3, H3K27Ac, and H3K27me. RESULTS/ANTICIPATED RESULTS: PDOs were established in 19 of 20 (95%) attempted cases. BAP1 protein expression was retained (n=7) or lost (n=12) in the primary tumors, with matching phenotype confirmed in PDOs. In 9 sequenced cases, a driving mutation was present in GNAQ (n=4), GNA11 (n=4), or CYSLTR2 (n=1). Morphology ranged from spindle-like to epithelioid clusters, mimicking primary tumor histopathology. Pigmentation increased with time in culture. Growth in culture was slow, and 1-2 months were allotted prior to passaging in most cases. Whole exome and RNA-sequencing confirmed distinct molecular profiles, with differential staining of active chromatin marks by IHC. DISCUSSION/SIGNIFICANCE: A biobank of primary UM PDOs with unique morphological and molecular characteristics has been established. These will serve as a model of human disease to facilitate translational research and investigate personalized treatments for patients with UM.

OBJECTIVES/GOALS: Pharmacogenomic testing for major depressive disorder is an expanding area of precision medicine with multiple direct-to-provider tests available. While clinical evidence supporting testing is expanding, there has been little research on the views and experiences of patients and clinicians utilizing this novel intervention. METHODS/STUDY POPULATION: This ongoing study is conducting semi-structured interviews with clinicians and patients exploring their views of the benefits and limitations of pharmacogenomic testing. Qualitative interviews have been conducted with 10 patients and 10 clinicians who have experience with ordering or receiving results within the past 12 months. Interviews are being thematically coded following a modified grounded theory approach using the Dedoose software. Following the principles of exploratory sequential mixed methods design, findings will be used to develop a survey to be administered to prescribing clinicians in both primary care and psychiatry. The survey will examine clinician's knowledge, interest, and concerns about utilizing testing. RESULTS/ANTICIPATED RESULTS: Preliminary analysis of qualitative interviews indicates that both patients and clinicians find that the broader testing process has benefits beyond the test results themselves. Benefits identified by patients include an increased trust in the process of selecting medications, validation of their negative experiences with medications, and improved communication with their provider. Limitations identified by patients include difficulty in accessing test results, and gatekeeping for testing by providers. Benefits identified by clinicians include increased empathy with patients, medication adherence, and improved communication with patients about medication. Limitations identified by clinicians include difficulty with ordering and interpreting test results. DISCUSSION/SIGNIFICANCE: Medication selection is a difficult process for both patients and clinicians. Improvements to clinician-patient communication and medication adherence are important benefits to consider in the adoption of testing. Future research should include these dimensions in assessment of the benefits and limitations of testing.

OBJECTIVES/GOALS: We leverage the disparate incidence of Ewing sarcoma (ES) between European (EUR) and African (AFR) ancestry to study ES tumorigenesis in iPSC-derived cells from donors with a range of AFR ancestry via functional / molecular profiling. Integrated multi-omics analysis furthers explore local regulatory networks in pursuit of novel drug targets of ES. METHODS/STUDY POPULATION: In our pilot, eight induced pluripotent stem cell lines were obtained, differentiated into neural crest cells, and then transduced with a lentivirus expressing GFP-2A-EWS/FLI1. We compared wild type (WT) to EWS-FLI1-induced cells and then compared cell survival, gene expression, and EWS-FLI1 binding differences at varying levels of EUR / AFR ancestry admixture. We will build on this pilot data by expanding the number of cell lines and measuring chromatin state. Subsequently we will refine our understanding of the relationships between local ancestry, epigenetic and gene expression changes, and phenotype in tumor progression via integration of multi-omics datasets. Our systems genomics approach will utilize directed local regulatory networks in a Bayesian structure learning framework. RESULTS/ANTICIPATED RESULTS: Induction by EWS-FLI1 resulted in gene expression changes enriched in known ES gene sets. Higher %EUR ancestry correlated with prolonged maintenance of EWS-FLI1. We identified thousands of ancestry-linked changes to gene expression and EWS-FLI1 binding. Eighty of these genes are both differentially expressed and differentially bound based on AFR ancestry admixture level and may be some of the early critical targets that initiate the cascade of molecular changes in ES. We will identify novel drug targets, with potential cross functional use of known drugs. Once we have developed directed local regulatory networks, we will use them to test in silico potential perturbations due to small molecules or novel drugs and predict expression changes. DISCUSSION/SIGNIFICANCE: With a limited number of cell lines, we identify 80 ancestry-linked candidate loci for functional validation through genome engineering. As EWS-FLI1 itself has proven elusive to direct targeting, studying its immediate downstream effects has the potential for establishing new druggable biologic pathways for treatment of ES.

OBJECTIVES/GOALS: Familial Hypercholesterolemia (FH) is a common disorder that is vastly underdiagnosed and causes an increased risk for sudden cardiac death. Cardiology providers (CHCPs) are in an ideal position to care for patients with FH. This research aimed to understand the practice behaviors of CHCPs in the screening, diagnosis, and management of FH. METHODS/STUDY POPULATION: An explanatory mixed methods design was utilized for this study. Adaptation of an existing FH knowledge tool guided survey development. The results of the quantitative survey, along with the Knowledge to Action framework and Theory of Planned Behavior, guided development of the interview protocol. Convenience and snowball sampling recruited CHCPs in the Division of Cardiology at Columbia University Irving Medical Center (CUIMC). Descriptive statistical analysis was performed on survey data. Qualitative interviews were conducted with survey respondents who volunteered to participate. Interviews were audio recorded, transcribed, and analyzed thematically. A descriptive review of the educational materials offered by the Division of Cardiology was conducted to identify FH knowledge domains presented. RESULTS/ANTICIPATED RESULTS: CHCPs with MDs, at CUIMC for 6-10 years, in clinical practice for 1-5 years, and in Inpatient Services had the highest average total FH knowledge scores. CHCPs with RNs, at CUIMC for less than 1 year, in clinical practice for 6-10 years, and in Cath Lab had the lowest average FH knowledge scores. Twenty interviews were completed, and four themes emerged- variability in FH care; issues related to addressing FH at institutional, practice setting and individual levels; importance of identifying FH early; and intervention approaches to overcome barriers to caring for FH patients in cardiology. CHCPs with MDs or with experiential FH knowledge were the only CHCPs to describe FH care beyond the point of screening. The document review revealed that only MDs were provided four lectures over the course of 4 years pertaining to FH. DISCUSSION/SIGNIFICANCE: CHCPs with didactic or experiential FH knowledge provided care beyond screening. Future interventions should increase didactic and experiential FH knowledge by incorporating institutional, local, and national FH resources. Improving the FH care CHCPs provide, can reduce FH-related morbidity and mortality as well as improve FH health outcomes.

OBJECTIVES/GOALS: The effect of the Direct-Acting Antiviral (DAA) on hepatic histopathological features from patients treated for HCV has not been thoroughly evaluated. The goals of this retrospective study were to determines differences between the liver biopsies collected before and after DAA treatment and correlated the histopathology with clinical outcome. METHODS/STUDY POPULATION: Spectral imaging was used to evaluate differences in intrahepatic macrophage (CD68, CD14, CD16, MAC 387, and CD163) and T cell (CD3, CD4, CD8, CD45, and FoxP3) phenotypes in paired liver biopsies collected from the same patient before (n=10) and after (n=10) achieving SVR (Figure 1). Imaging analysis and machine learning algorithms were used to evaluate changes in these key immune cells. We also compared differential gene expression of over 700 genes using RNA isolated from liver biopsies with NanoString. RESULTS/ANTICIPATED RESULTS: Multispectral imaging analysis showed a significant increase of proinflammatory/M1-like (e.g., CD14+) and anti-inflammatory/M2-like macrophage (e.g., CD163+) phenotypes in pre-treatment versus post-treatment biopsies, respectively. Gene expression analysis revealed enrichment of inflammatory (HLA-B, STAT1, CXCL10) and interferon induced-antiviral (ISG15, OAS3, MX1 and IFIT1) genes in the pre-treatment vs the post-treatment group. Cell deconvolution analysis also showed a significant increase of M1-like macrophages in the pre-treatment group when compared to the post-treatment group or controls. Upregulation of genes associated with cell proliferation and differentiation (c-KIT and Fos) was observed in the post-treatment biopsies of patients with persistent inflammatory infiltrates. DISCUSSION/SIGNIFICANCE: Protein and gene expression profiles observed in patients before DAA therapy showed a strong macrophage-mediated inflammatory response against HCV infection in the liver that shifted significantly to a tissue remodeling microenvironment after treatment.

OBJECTIVES/GOALS: In this study, the ability of a pathologist to detect malignancy on digital pseudo-H&E slides obtained via structured illumination microscopy (SIM) imaging of fresh diagnostic breast biopsies was assessed. The speed of imaging and processing was also assessed for potential clinical implementation. METHODS/STUDY POPULATION: This study was conducted in accordance with an Ochsner Medical Center of New Orleans IRB. 200 patients undergoing either stereotactic or ultrasound-guided diagnostic breast biopsies were consented and an additional core from the suspicious lesion was collected for research use. Research biopsies were transported to the lab and stained with DRAQ5 and Eosin-Y and imaged with SIM before being submitted for histology processing. Imaging and digital processing times were recorded. The resulting SIM images and histology slides were given to a pathologist for blind review to assess accuracy. RESULTS/ANTICIPATED RESULTS: The ex-vivo structured illumination microscopy images and subsequent histology slides from 79 research cores have been assessed to date. Some samples were excluded from the total data set and not included in the final assessment due to technical failures of the imaging protocol. Of the current set, the pathologist has a specificity of 88% and a sensitivity of 65%, as well as an NPV of 88% and a PPV of 65%. Staining time for each biopsy was completed within 3 and a half minutes and imaging at 20x magnification took between 4 and 12 minutes, depending on size and implementation of autofocus to the imaging system. Image processing took approximately 5 minutes per biopsy and is a direct function of biopsy size. DISCUSSION/SIGNIFICANCE: Decreased time between cancer suspicion and treatment will improve the prognosis of breast cancer patients. SIM imaging of fresh breast biopsies could ultimately allow primary and secondary histology to be performed simultaneously and minimize histopathology time, thus allowing clinicians and patients to implement treatment course more quickly.

OBJECTIVES/GOALS: IgA1 nephropathy, which can lead to kidney failure, is caused by complexes formed between aberrant galactose-deficient IgA1 and antibodies directed against it. Our goals are to characterize shifting glycosylation patterns at the level of single IgA1 molecules and to apply this to patient samples for early detection and understanding of the disease. METHODS/STUDY POPULATION: To characterize glycosylation patterns on single IgA1 molecules, labelled IgA1 in low concentration was physisorbed to borosilicate glass in a fluidic cell and labelled Jacalin was flowed in to bind with Glycans on IgA1. The samples were observed with a Nikon TiE epi-fluorescence microscope. FRET images were created by exciting the Jacalin dye with a blue laser and recording the red emission of the IgA1 dye with an EMCCD camera. FRET emission intensities of individual IgA1 molecules over time were analyzed to determine how frequent and how long Jacalin binds to each of them. The rate of binding is roughly inversely proportional to the amount of abnormal glycans on a given IgA1 molecule. After the method is perfected, we intend to compare the glycosylation patterns of healthy and diseased patient samples. RESULTS/ANTICIPATED RESULTS: Addition of the competitive binder Galactose to the solution led to an increase in the off times of Jacalin and IgA1 and a decrease in the on times in a concentration-dependent manner, yielding an increase in the dissociation constant. Dissociation constants of individual molecules within a single experiment vary by 3 orders of magnitude, which cannot be attributed to stochastic fluctuations but rather reflects differences in the adsorption geometry. Nevertheless, the unaffected dissociation constant can be identified. We expect that when this method is applied to samples from healthy and IgA1 Nephropathy patients, specific IgA1 molecules from patients will have higher dissociation constants for Jacalin compared to those from healthy patients. DISCUSSION/SIGNIFICANCE: The binding rates of Jacalin to single IgA1 vary by 3 orders of magnitude. The observed heterogeneity shows the Jacalin probe can differentiate between different IgA1 populations. An understanding of which IgA1 molecules in patient samples are problematic and what their distribution of Glycans is can lead to discovering biomarkers and treatments.