There is growing emphasis on the importance of active partner engagement in research and the design, planning, and implementation of new programs. Indeed, the science of engagement is gaining is receiving more attention from researchers, being formally included in research studies more often, and increasingly being required by funders. Financial costs and economic implications are well-known drivers of successful adoption, implementation, and sustainment of evidence-based programs; however, partner engagement costs are often excluded from economic evaluations even when elements of engagement may be relevant to sustainment and replication. Resource constraints in both research and practice highlight the need to better understand engagement costs to ensure that scarce resources are put towards the richest engagement activities. As part of the Rapid and Rigorous Patient-Centered Program (R2P2) in Colorado, we designed an approach to systematically document and analyze partner engagement costs across four prototype projects. We describe this approach, summarize needed adaptations, share an interactive data collection template, synthesize the relevant engagement costs during implementation, replication, and sustainment, and discuss lessons learned. Our experiences in R2P2 in costing initial engagement activities across four diverse projects offer direction for future research and practice that balances high-quality partner engagement with efficiency in addressing resource constraints facing researchers and practitioners.

OBJECTIVES/GOALS: Lack of diversity within clinical trials is well known, but there is little data on the use of investigational products through other pathways, such as expanded access. This project sought to determine the demographic diversity of patients benefiting from expanded access at Michigan Medicine. METHODS/STUDY POPULATION: Previous quality improvement reviews provided the list of 271 patients for whom a single-patient, expanded access request had been approved by the FDA and University of Michigan IRBMED between 2005 and 2021. Demographic information was collected through the EMERSE tool, including age, legal sex, race, ethnicity, and zip code. These data were cross-referenced with available regulatory documentation on product requested, treatment area, and date of request. Descriptive statistics were performed using Excel. RESULTS/ANTICIPATED RESULTS: Patients who were approved to use an investigational product through expanded access at Michigan Medicine showed a wide geographic distribution, including 48 Michigan counties, 20 states, and 1 province. All age groups were served, with those between 30-49 underrepresented and those under 10 and over 60 overrepresented. Race data generally followed the proportions of the Michigan state census, including 76% white and 14% black or African American (expected: 79% and 14%) and 48% female (expected: 50%). On further breakdown, populations differed by specialty and county. DISCUSSION/SIGNIFICANCE: The distribution of Michigan Medicine patients with approved expanded access requests was similar to the population of Michigan with respect to age, sex, and race. Further research is needed to determine if this reflects equitable use or if these results are generalizable to other institutions.

OBJECTIVES/SPECIFIC AIMS: We intended to investigate the relationship between children’s genetic admixture proportions and the incidence of obesity at 12 months of age METHODS/STUDY POPULATION: 821 twelve month old children were included and their genetic admixture was estimated using the ancestry and kinship toolkit (AKT). Weight for Length Percentile (WFLP) at 12 months of age was categorized as <95th percentile and ≥95th percentile. Multiple logistic regression analysis was performed to calculate odds ratios (ORs) with 95% confidence intervals (CIs) for association of admixture proportion including EUR, AMR, AFR, EAS and SAS with WFLP categories adjusting for maternal education, birth weight, frequency of breast feeding and juice consumption RESULTS/ANTICIPATED RESULTS: 821 children were included; WFLP <95th percentile = 671 (81.7%) and WFLP ≥95th percentile = 150 (18.3%). Crude odds ratios showed EUR admixture was protective (OR 0.45 (95% CI 0.27–0.74)) whereas AMR (OR 3.85 (95% CI 1.92 –7.70)) and AFR (OR 5.70 (95% CI 2.19 –14.85)) were positively associated with obesity. After adjusting for confounding variables, only AFR was associated with WFLP≥95th (OR 7.38 (95% CI 2.31 – 23.59)), AMR and EUR were no longer associated with WFLP≥95th DISCUSSION/SIGNIFICANCE OF IMPACT: The AFR admixture proportion remains associated with early childhood obesity after accounting for confounding variables, suggesting the ancestral genetic background may contribute to the differences seen in early childhood obesity

Objectives/Goals: Healthcare organizations must track electronic health record (EHR) activity at the user level, including logons, accessed records, and viewed or entered documentation. There is little standardization in EHR audit logs and nurse workload has not been explored using these data. In this project, we characterized nurse actions from EHR audit logs. Methods/Study Population: We performed an analysis of EHR audit log data collected from 8,149 nurses over 5 years at University of Utah Health. We preprocessed nursing-centric EHR audit logs from the Epic EHR by cleaning and preparing the data for analysis. We calculated basic statistics for the variables labeled user_id (nurse) and metric_id (action). We reviewed the actions used by nurses and categories the actions as navigation, view, and entry. To capture the clinical context of the actions, two nurses categorized each action. A third nurse resolved any discrepancies. Results/Anticipated Results: We found that of the 4,419 available metrics, nurses used 1,461 unique metrics during the timeframe. The actions most used by nurses were 1) report with patient data viewed, 2) inpatient system list, and 3) storyboard viewed. Most of the metrics were categorized as navigation. The number of nurses interacting with the EHR increased each year and on average, we found that 1000 unique metrics were used by each nurse user in a 24-hour period. The expected outcome is a set of actions that can be mapped to higher level nursing interventions and in the future contribute to models for nursing workload measurement. Discussion/Significance of Impact: We found great value in using EHR audit logs to provide insights into nursing actions. Information gleaned can benefit organizations that are crafting interventions to decrease workload. Ultimately, the goal is to ensure that nurses have an appropriate workload allowing for safe and high-quality patient care while maintaining their well-being.

OBJECTIVES/SPECIFIC AIMS: The aim of the present study is to explore the relationships between resilience, pain catastrophizing, and functional outcomes in pre-and post-operative TKR patients. The primary outcome is the ability of the Brief Resilience Scale (BRS) to predict function using the Knee injury and Osteoarthritis Outcome Score Joint Replacement (KOOS, JR.) and overall health on the Patient-Reported Outcome Measurement Information System Global Health (PROMIS GH). METHODS/STUDY POPULATION: Recruited patients will provide informed consent during in-person office visits. At the pre-surgery intake appointment, patients will provide demographic information and complete baseline questionnaires assessing resilience, pain catastrophizing, knee stiffness, pain, and function, and impression of overall health. In-person or electronic follow-up assessments will be administered post-surgery at 6-week, 3-month, and 6-month appointments. RESULTS/ANTICIPATED RESULTS: We aim to recruit 150 patients for this study. We do not expect a difference in sociodemographic characteristics of the patient sample. P-values will be based on t-tests and correlations calculated by comparing only non-missing values. Each outcome measure will be examined at each time point and trajectories will be calculated to determine the change in each outcome over time. Using latent class growth modeling we will examine individual change over time for each outcome (BRS, PCS, PROMIS GH, and KOOS JR.), and the association of resilience and pain with change in function (KOOS, JR. total score) and overall health (PROMIS GH). DISCUSSION/SIGNIFICANCE OF IMPACT: To our knowledge, this is the only known study that will compare resilience, pain catastrophizing and longitudinal health outcomes in a single paradigm. Results will hopefully inform the design and implementation of future studies that will assess the impact of preoperative treatment of vulnerable patients. The ultimate goal is to improve functional recovery by using pre-identified psychological constructs as intervention points. Ideally, the same measures would be implemented, however, details of this plan will be established following completion of this study.

OBJECTIVES/GOALS: The goal of this study is twofold: To develop a method for an ex vivo hypofibrinolytic control and second to analyze patterns in standard and recently developed clinical coagulation assays for the detection of hypofibrinolytic states. METHODS/STUDY POPULATION: We analyzed blood samples from healthy patients first under normal conditions and then laced with human recombinant PAI-1 under three different concentrations. We then analyzed both samples using standard clinical assays (PT, aPTT, D-dimer, Fibrinogen), thromboelastography point-of-care tests (Hemosoncs- Quantra system), and with research assays of clot size and aggregation. Our previous research of diagnostic errors showed the patient group with the highest overall risk of these non-identifiable thrombotic complications was post-menopausal women with chronic diseases. We therefore focused our patient population to healthy post-menopausal women who were not using hormone replacement therapy. RESULTS/ANTICIPATED RESULTS: Research assays showed PAI-1 significantly increased clot size and aggregation. Preliminary results of clinical assays showed no detectable difference in hypofibrinolytic samples at any concentration. We anticipate ongoing testing will show similar results. Results on Quantra tests showed much larger differences between control and hypofibrinolysis samples, and we anticipate ongoing testing will achieving statistical significance. It is still unknown whether the mean value for hypofibrinolysis samples on the Quantra Clot Stability assay will be outside of the “normal” reference range. We theorize that this may be due to hypofibrinolytic changes in the overall structure and core density of the clots. DISCUSSION/SIGNIFICANCE: Cellular stress stimulates a concomitant activation of inflammation and coagulation, including decreased fibrinolysis. Unfortunately, current clinical assays do not assess clot breakdown. This connection would account for the increased rate of thrombosis in patients with chronic inflammation without detectable results on clinical tests.

OBJECTIVES/GOALS: Suicide is the second leading cause of death in teens. Substance misuse, including opioids, is a risk factor for suicidal thoughts and behaviors among teens. School-based approaches are promising, yet they lack evidence on their effectiveness. To fill this gap, we created supportED, a digital game to prevent suicide in youth who misuse substances. METHODS/STUDY POPULATION: We conducted focus groups and interviews to inform the development of the digital game and discuss approaches to enhance user experiences, acceptability, and feasibility of the intervention. A semi-structured focus group/interview guide was developed, pilot tested, and used in focus groups/interviews. The guides aligned with constructs from a safety planning intervention, a well-recognized, evidence-based suicide prevention intervention, to identify potential warning signs of emotional distress, coping strategies, and resources for seeking help to prevent suicidal thoughts and behaviors. Focus groups/interviews were led by a facilitator, who is also a certified school counselor and licensed professional counselor, and a co-facilitator who took field notes and provided debrief summaries. RESULTS/ANTICIPATED RESULTS: A total of 35 participants, including 10 high-school-aged teens [aged 16-19], 10 college-aged youth [aged 18-22], 10 school-based providers, and five interviews with adults who had experience with opioids in their youth informed the development stage. Of the focus group participants, 25.7% identify as female (n=23), 71.4% identify as male (n=6), and 2.9% do not identify as male or female (n=1); 60% identify as White (n=18), 20% identify as multi-racial (n=6), 10% identify as Black (n=3), 3.3% identify as Asian (n=1), and 6.7% identify as race unknown (n=2); and, 37.1% (n=13) identify as Hispanic/Latinx. Salient themes that emerged focused on opioids, substance misuse, mental health, bullying, grief, family, identity, and stigma to create six distinct narratives in the digital game. DISCUSSION/SIGNIFICANCE: Findings informed the development of a digital game to prevent suicide among adolescents who misuse substances. The digital game is currently being evaluated in a pilot RCT to assess the preliminary efficacy of the intervention, user experience, usability, and feasibility of delivering the intervention and protocols among 60 youth.

Objectives/Goals: The impact of genetic alterations on survival in patients with neuroendocrine prostate cancer (NEPC) is unclear. This study assessed how genetic alterations and tumor mutational burden (TMB) affect overall survival (OS) in patients with NEPC treated at Mayo Clinic. Methods/Study Population: Demographic, clinicopathologic, and genomic data from patients with NEPC treated at Mayo Clinic (2015–2025) were abstracted from electronic medical records. Patients had consented to research and received at least one cycle of therapy. Descriptive statistics summarized patient characteristics, and survival was estimated with Kaplan–Meier analysis. OS was compared by disease type (de novo vs treatment-emergent NEPC), TMB (high vs low), and RB1, TP53, and PTEN alteration status. Results/Anticipated Results: Among 66 patients in the study, the average age at NEPC diagnosis was 67.5 years. There were no significant differences between the age at diagnosis of de novo and treatment emergent NEPC (t-NEPC). t-NEPC made up 77.3% of the cohort. Median follow-up was 46.5 months (IQR 28–120). The most common alteration was TP53. RB1 alterations were associated with significantly shorter OS compared to patients without RB1 alterations (mOS Not Reached (NR) vs 13.8 months; p = 0.0056). Similarly, patients with high TMB had worse overall survival (mOS NR vs 14.9 months; p = 0.077). We did not observe worse OS in patients with alterations in TP53 or PTEN. RB1 alteration and high TMB were the strongest predictors of worse survival. Discussion/Significance of Impact: RB1 alterations and high TMB predict poor survival in patients with NEPC, highlighting potential biomarkers for risk stratification. TP53 and PTEN did not significantly affect OS. Early genomic profiling and targeted management of actionable alterations such as BRCA and ATM may improve survival and inform future translational studies.

Objectives/Goals: The Human Immune Dysregulation Evaluation Framework (Hi-DEF) quantifies myeloid dysregulation in infectious and non-infectious illness using peripheral blood gene expression (Moore, Nat Med). We evaluated immune dysregulation in obesity and quantified the changes in dysregulation due to lifestyle and medical interventions. Methods/Study Population: We assessed the association of obesity with myeloid dysregulation in the Framingham Cohort using linear regression. In two additional datasets (GSE66175, GSE19790), we assessed the effect of dietary interventions and bariatric surgery on myeloid dysregulation using paired Wilcoxon rank sum test pre- and post- intervention and Pearson’s correlation. Results/Anticipated Results: In the Framingham cohort (n=5321), myeloid dysregulation score was associated with BMI (p=2.2e-6) after correcting for age and sex. Patients assigned to diet/exercise experienced significant reductions in myeloid score after 3 months (n=63, p=1.2e-7), which was not seen in controls (n=63, p=0.29). Change in myeloid score was significantly correlated with change in weight loss (r=0.32, p=0.006). This reduction in myeloid dysregulation was also seen post-bariatric surgery (n=11, p=0.0001). Discussion/Significance of Impact: Patients with obesity have significantly higher myeloid dysregulation compared to healthy subjects, which is modifiable with lifestyle and medical interventions. These results suggest that we may be able to quantify and monitor immune dysregulation from obesity which may place these patients at risk for worse outcomes.

OBJECTIVES/SPECIFIC AIMS: Our objective is to examine the relationship between depression, childhood asthma and family structure. METHODS/STUDY POPULATION: We analyzed data from The National Survey of Children’s Health 2011-2012 for the variables family structure, parent’s smoking, education, poverty, available emotional support, and children’s age, gender, race/ethnicity, asthma, physical/mental health, access to care, and insurance. Data were analyzed using multiple logistic regression in STATA 14 (sampling and weight). RESULTS/ANTICIPATED RESULTS: Of the 95,000 participants, 3.8% had depression, 5.1% had anxiety, 8.5% had emotional, developmental, or behavioral problem, 14.6% had asthma and 19% were single-parent family. Asthmatics had higher chance for depression relative to non-asthmatics (Adjusted Odds Ratio (AOR)=1.5, 95% confidence interval (CI)=1.2-1.8), p=0.001). Those living in a single-family home had higher chance for depression than those living in 2-parents homes (AOR=2.5, 95% CI=1.9-3.2), p=0.001). Other predictors were lack of emotional support, age, poverty <100%, poor health, and smoking parent (p<0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: We concluded that depression was associated with asthma in children from poor single family who lack an emotional support. Management of asthma should include screening and management of psychosocial factors.

ABSTRACT IMPACT: This study is providing a telehealth intervention for the first time in patients with sickle cell disease with the goal of improving cognitive functioning and preparing adolescents for successful transition of care to adult healthcare providers. OBJECTIVES/GOALS: There is a high prevalence of cognitive impairment in adolescents with sickle cell disease (SCD). The purpose of this study is to test the efficacy of an individualized cognitive remediation program designed to promote not just cognitive function, but also adaptive and self-management skills necessary for successful transition to independence. METHODS/STUDY POPULATION: 12 participants with SCD (5 males, ages 10-16) participated in an individualized program, Cognitive-Remediation of Executive and Adaptive Deficits in Youth [C-READY], consisting of three main components: individual goal-based therapy, parent training sessions, and home skill practice. C-READY sessions occur one-on-one with a trained therapist for 8 sessions conducted over 4 weeks. Weekly parent training sessions are also conducted as part of the C-READY program. All of these sessions occurred via telehealth video-calling between the therapist and the adolescent/parent. Participants were evaluated before and after the C-READY program using neuropsychological assessment measures and transition readiness questionnaires. Parents also completed ratings on telehealth delivery, content, and timing. RESULTS/ANTICIPATED RESULTS: Repeated measures ANOVA indicated significant improvement in transition readiness behaviors as rated by parents, including improved independence in medication management (p = 0.029) and in talking with their healthcare providers (p = 0.019). Significant improvement was also demonstrated on a neuropsychological measure related to executive function skills, specifically inhibition and switching (p = 0.012). Results from telehealth surveys (rated on a 5-point Likert scale) indicated overall satisfaction with services (4.2/5), including visual (4/5) and voice quality (4.3/5) of telehealth equipment. Ratings also indicated feeling that their privacy was respected (4/5) and that their interactions with their therapist were appropriate and sensitive (4.5/5). DISCUSSION/SIGNIFICANCE OF FINDINGS: These results provide support for interventions that focus on cognitive skills to improve behaviors necessary for successful transition of care in youth with SCD. Results are also promising for delivery via telehealth in order to address barriers related to access to care. Future results will continue to be reported, as this study is currently ongoing.

Objectives/Goals: Despite the documented link between ultraviolet (UV) exposure and skin cancer, the potential protective role of pigmentation in preventing skin carcinogenesis is not fully understood. This project will test the hypothesis that individuals with light skin tones are more susceptible to skin cancer due to a higher accumulation of somatic mutations from UV exposure. Methods/Study Population: Skin biopsies were collected from various anatomical sites of seven donors aged 70 to 80 years with dark skin tones and used to generate clones of normal skin cells. The clones were then subjected to whole-exome and RNA sequencing. We developed a workflow that allows us to accurately detect somatic mutations in clonal expansions of individual cells with high specificity and sensitivity. For comparison, we also analyzed mutations in skin cells obtained from individuals with light skin tones, who served as our control group. In the future, we plan to employ admixture analysis to clarify the impact of race on somatic mutations by examining ancestral genetic contributions. Results/Anticipated Results: Our preliminary analysis of the existing data supports our hypothesis. The average mutation burden in individuals with light skin tone was found to be 3.778 Mut/Mb, whereas in those with dark skin tones, was only 0.818 Mut/Mb. These findings align with the prevailing hypothesis that melanin acts as a protective factor against skin cancer, as it appears to reduce the mutagenic effects of UV radiation. This suggests that individuals with darker skin may have a lower cumulative mutational load, potentially contributing to their reduced risk of developing skin cancers such as melanoma. Similarly, our results showed variations in mutational burdens across different anatomical sites, which seemed to be influenced by differing levels of UV exposure, with the highest burdens observed in areas with the greatest exposure. Discussion/Significance of Impact: Understanding how skin cancer operates in darker-skinned individuals is imperative for tailoring effective screening and targeted therapies to meet their specific needs. In the long run, elucidating the mechanisms underlying skin cancer development in this demographic will help refine screening protocols and prevention recommendations.

OBJECTIVES/GOALS: To determine whether length of stay (LOS) and opioid prescribing differ among patients who present to the emergency department (ED) with low back pain (LBP) and serious mental illness (SMI+) compared to patients without SMI (SMI−). METHODS/STUDY POPULATION: Eligible patients that visited the ED within the Mount Sinai Health Care System from 2016-2019 were identified from the Mount Sinai Data Warehouse. Data on patient demographics, number of medications prescribed, and length of stay (LOS) were compared between the groups. Patients were excluded if English was not their primary language and if the LOS exceeded 24 hours. The final dataset consisted of 940 patients (SMI+: n = 181; SMI−: n = 759). RESULTS/ANTICIPATED RESULTS: SMI+ cases included patients with a diagnosis of depression (n = 152), anxiety (n = 134), schizophrenia (n = 9), bipolar (n = 1), and/or post-traumatic stress disorder (n = 33); 26% of cases had a single diagnosis, 66% with two, and the remaining 8% had three diagnoses. There was no significant difference in pain scores between the two groups (SMI-: 7.0 ± 0.1; SMI+: 6.8 ± 0.3; p = 0.6). We found no significant differences in LOS between the groups (SMI-: 3.9 ± 0.1 hours; SMI+: 3.8 ± 0.2 hours; p = 0.8), nor was there a significant difference in number of medications prescribed (SMI-: 1.7 ± 0.9; SMI+: 1.7 ± 0.6; p = 0.4). Further analysis revealed that the odds of receiving an opiate prescription in the SMI- group was 0.92 (95% CI: 0.54,1.55). DISCUSSION/SIGNIFICANCE OF IMPACT: Comparable opioid prescribing and LOS exist in patients with and without serious mental illness who are seeking treatment for low back pain in the ED. Despite similarities in approaches to care, more information is needed to determine if other social determinants influence these practices.

OBJECTIVES/GOALS: Vanishing White Matter Disease (VWM), is a childhood neurodegenerative leukodystrophy that presents with motor deficits, neurologic decline, and seizures leading to death.There are no treatments. Herein we investigate adeno-associated virus serotype 9 (AAV9) gene addition therapy for VWM. METHODS/STUDY POPULATION: To serve as a baseline for disease correction, we characterized the severe VWM Eif2b5I98M murine model with clinically relevant readouts including motor function, gait mapping and myelin loss through magnetic resonance imaging (MRI). Molecular characterization through the identification of biomarkers was also investigated. To provide targeted disease correction, we designed four gene replacement constructs to drive the rapeutic EIF2B5 expression in astrocytes—a critical cell type for VWM pathology. We are currently evaluating our AAV vectors in two murine VWM models, Eif2b5R191H and Eif2b5I98M, and are monitoring disease progression using traditional and clinically relevant readouts. RESULTS/ANTICIPATED RESULTS: The I98M mice display significant mobility loss, ataxic gait, and demyelination. Molecular characterization also indicates that the integrated stress response is significantly dysregulated, supporting the classic VWM phenotype. Our previous biodistribution study confirmed our ability to efficiently target astrocytes using varying iterations—including one novel—of the glial fibrillary acidic protein (GFAP) promoter. Our data suggests that targeting astrocytes with gene addition delays disease onset, partially rescues motor function, and attenuates myelin loss. Survival of the AAV9-gfaABC(1)D-EIF2B5 treated I98M mice is also significantly increased (p<0.0001), currently with a 2-fold extension in life expectancy. DISCUSSION/SIGNIFICANCE: Overall, we anticipate emergence of a lead astrocyte-targeted gene therapy candidate in which the data will be strengthened through the evaluation of clinically relevant measures in two murine models of disease, allowing fortimely translation to the clinic.

OBJECTIVES/GOALS: The purpose of the project was to create a Tableau dashboard to track metrics on requests for research data at Atrium Health Wake Forest Baptist. The objectives included: 1) define and identify request fulfillment metrics, 2) build a dashboard to capture metrics, and 3) integrate the dashboard into metrics tracking and reporting activities. METHODS/STUDY POPULATION: Project managers and team leaders in the Office of Informatics collaborated to determine which measures would be most relevant and impactful to report on. Metrics that were collected included: total count of tickets fulfilled over time, number of tickets currently open, sum of outstanding quoted hours, quoted hours vs. actual hours needed to fulfill ticket, and hours billed. Tableau's direct connection feature was used to extract the Trac ticket data from its Postgres database and the dashboard was published to Tableau Server. After the initial draft was created, several rounds of revisions were made as new data insights were discovered through further investigation of the data. RESULTS/ANTICIPATED RESULTS: Each morning, Tableau Server runs an automatic refresh of the data. On the dashboard homepage, users can see a quick view of all available metrics; to minimize noise, only the current statuses, active tickets, and stats for the most recent monitoring periods are displayed. Many of the charts give the user the option to link out to a page with related supplemental information (historic data, ticket status history, etc.). With the help of the dashboard, project managers and team leaders can now monitor how long tickets are in each status, increase quote accuracy using the hours quoted and hours billed charts, and examine ticket complexity over time. DISCUSSION/SIGNIFICANCE: Prior to dashboard creation, metrics were sparse and difficult to assemble. By providing information on the quantity, size, and complexity of data requests, the dashboard enables the Office of Informatics to monitor how the process is functioning overall, make informed decisions about resource allocation, and provide quick interventions.

ABSTRACT IMPACT: Building a patient cohort to support clinical-translational research in chronic hepatitis B OBJECTIVES/GOALS: The overall objective of my KL2 project is to delineate the effect of HIV coinfection, and CHB disease stage and hepatitis B (HBV) viremia on the ability of toll-like receptor 8 agonism to restore HBV-specific immune cell function. This abstract describes the characteristics of the cohort from which research blood samples for my KL2 are collected. METHODS/STUDY POPULATION: HOPE is a prospective cohort study enrolling people with CHB including HIV/CHB coinfection, and resolved CHB. Participants are enrolled at primary care clinics in Maryland, Washington D.C., and Virginia. Standard-of-care antiviral treatment with tenofovir alafenamide (TAF) is prescribed through the study if indicated. Patients receiving TAF from the study are either starting treatment, or switching to TAF from another antiviral medication. If receiving TAF, participants are seen every 3-6 months for medication refills, clinical and research blood draws, and adverse event evaluations. Liver fibrosis is measured by FibroScan and a minority undergo liver biopsy. RESULTS/ANTICIPATED RESULTS: So far, 204 people have been enrolled, 177 with CHB (23 HIV/CHB coinfection), and 22 with resolved HBV infection. To date, 45 patients who were viremic at baseline and initiated on TAF have been enrolled. CHB predominantly affects Asian and African immigrants in the U.S, and the majority (77%) of HOPE participants are immigrants from these countries. The majority are male (70%), mean age 51 years (SD ±14). So far, 86 people with CHB monoinfection have been prescribed TAF on-study. Thirty-five percent of these had significant (>F2) liver fibrosis at baseline and half had elevated ALT (mean 47.5, SD ±55 IU/ml). Forty-three percent switched to TAF from another oral antiviral. Most switched due to lack of coverage by health insurance. DISCUSSION/SIGNIFICANCE OF FINDINGS: HOPE is a CHB cohort dedicated to collecting research samples and providing antiviral treatment. It is the foundation for the CHB translational research program at the University of Maryland School of Medicine. The availability of paired viremic and virally suppressed, HIV/CHB, and resolved HBV research samples are strengths of HOPE.

OBJECTIVES/GOALS: Antibiotic treatment sets the stage for intestinal domination by Candida albicanswhich is necessary for development of invasive disease, but the resources driving this bloom remain poorly defined. We sought to determine these factors in order to design novel prophylaxis strategies for reducing gastrointestinal (GI) colonization. METHODS/STUDY POPULATION: We initially developed a generalizable framework, termed metabolic footprinting to determine the metabolites C. albicanspreferentially uses in the mouse GI tract. After identifying the metabolites C. albicansutilizes, we usedin vitro growth assays in the presence and absence of oxygen to validate out metabolomics findings. We next determined if a probiotic E. coli that utilizes oxygen would reduce C. albicanscolonization compared to a mutant E. coli that could not respire oxygen. Finding that oxygen was a necessary resource, we utilized germ-free mice to determine if Clostridiaspp. known to reduce GI oxygen would prevent C. albicanscolonization. Lastly, we sought to see if 5-aminosalicylic acid (5-ASA) could prevent C. albicanscolonization. RESULTS/ANTICIPATED RESULTS: We found that C. albicans preferentially utilizes simple carbohydrates including fructo-oligosaccharides (e.g., 1-kestose), disaccharides (e.g., β-gentiobiose), and alcoholic sugars (e.g., sorbitol) and is able to grow in vitro on minimal media supplemented with either of these nutrients. However, in the hypoxic environment that is found in the “healthy” colon, C. albicans cannot utilize these nutrients. We next found that pre-colonization in a mouse model with a probiotic E. coli significantly reduced C. albicanscolonization, but the mutant E. coli had no effect on colonization. We next showed that Clostridia supplementation restored GI hypoxia and reduced C. albicanscolonization. Remarkably, we found that 5-ASA significantly reduced GI colonization of C. albicans. DISCUSSION/SIGNIFICANCE: We have shown that C. albicans requires oxygen to colonize the GI tract. Importantly, we found that 5-ASA can prevent an antibiotic mediated bloom of C. albicans by restoring GI hypoxia, which warrants additional studies to determine if 5-ASA can be used as an adjunctive prophylactic treatment in high risk patients.

Objectives/Goals: Colorectal cancer risk varies across populations. Bile acid (BA) composition and the gut microbiome can impact CRC risk, but their differences across populations is poorly understood. We characterized serum BA and gut microbiome profiles in non-Hispanic Black and White Americans and examined associations with diet and social determinants of health. Methods/Study Population: To characterize BA composition, participants from the Chicago Multiethnic Prevention and Surveillance Study with a banked serum sample (n=100) were selected, with a subset of participants having completed a diet recall survey (ASA24; n=23). Sixty-two BA (primary, secondary, and glyco/tauro-conjugated BA) from serum samples were analyzed using liquid chromatography-mass spectrometry. For microbiome profiling, normal colonic biopsy samples from non-cancer participants were obtained from the Digestive Disease Research Core Center’s Integrative Clinical and Biospecimens Core (University of Chicago; n=109). Following DNA extraction, the mucosally adherent microbial community was assessed by 16S rRNA sequencing. Results/Anticipated Results: Of the BA detected in over 80% of the samples, glycochenodeoxycholic acid (GCDCA) and deoxycholic (DCA) had the highest median concentrations (56.7 and 55.0 µg/mL, respectively). Moderate positive correlations between DCA/isoDCA and calories, fat, and cholesterol were observed. The linear regression model employed to understand the relationship between log-transformed DCA and race, age, sex, and area deprivation index highlighted age as a key predictor of DCA level when controlling for other covariates. Microbiome analysis revealed that diversity was not different when comparing sexes, races, or colonic anatomic locations. At the phylum level, taxa belonging to the Bacillota and Bacteroidota phyla were common across samples. Discussion/Significance of Impact: These data and subsequent analysis, including additional BA analysis, improvement of the regression model, and deeper analysis of microbiome community related to BA metabolism, will provide additional context to the role of social and biological factors in better understanding differences in CRC risk and pathogenesis across human populations.