OBJECTIVES/SPECIFIC AIMS: We recently reported that PRMT5 epigenetically activates androgen receptor (AR) in prostate cancer cells. Because targeting AR signaling through androgen deprivation therapy is clinically used as a radiosensitization approach to treat high-risk prostate cancer, our finding raised an exciting possibility that targeting PRMT5 may improve RT for prostate cancer patients. Contrary to our expectation, targeting PRMT5 sensitized both AR expressing and AR negative (AR−) prostate cancer cell lines to radiation. The goal of our study was therefore to determine the role of PRMT5 in repair of IR-induced DSBs and to translate these findings to improving radiation therapy for cancer patients in general (not just prostate cancer patients). METHODS/STUDY POPULATION: The majority of experiments were basic science experiments analyzing PRMT5’s role in the DNA damage response in normal and cancer cell lines. For example, to extend our findings and determine if PRMT5’s role in DSB repair is conserved across multiple cell types, we performed similar experiments in AR− prostate cancer cells, luminal breast cancer cells, glioblastoma cells, and human embryonic kidney cells. To determine the clinical significance of our finding, we also analyzed mRNA expression of PRMT5, AR, and both PRMT5 and AR target genes involved in DSB repair across 43 clinical cancer data sets. RESULTS/ANTICIPATED RESULTS: (1) Targeting PRMT5 sensitizes prostate cancer cells to IR in an AR-independent manner, (2) PRMT5 regulates the repair of IR-induced DSBs in an AR-independent manner, (3) RNA-seq analysis reveals that PRMT5 likely regulates genes involved in the DNA damage response, (4) PRMT5 activates expression of several genes in the DDR including those involved in DSB repair, (5) PRMT5 functions as an epigenetic activator of genes involved in DDR, (6) PRMT5 is required for NHEJ, HR, and G2-Arrest upon IR treatment, (7) Upregulation of PRMT5 correlates with formation and repair of IR-induced DSBs, (8) PRMT5’s role in repair of IR-induced DSBs is conserved in several normal and cancer cell types, and (9) PRMT5 expression correlates with expression of DSB repair proteins in clinical cancer samples. DISCUSSION/SIGNIFICANCE OF IMPACT: In summary, we provide evidence that PRMT5 is a master epigenetic regulator of IR-induced DSB repair through epigenetic activation of multiple target genes involved both HR and NHEJ as well as G2 arrest. Interestingly, the majority of genes regulated by PRMT5 are well-characterized, “core repair proteins” involved in HR (RAD51, BRCA1, BRCA2, RAD51D, and RAD51AP1), NHEJ (NHEJ1, Ku80, XRCC4, and DNAPKcs), and G2 arrest (Cdk1, CDC25C, CCNB2, and WEE1), which may explain why PRMT5 is essential to repair IR-induced DSBs in several cell lines. Although AR may also regulate DSB repair via both HR and NHEJ, several pieces of evidence in our study suggest that PRMT5 also regulates DSB repair independent of AR. First, PRMT5 targeting sensitizes both AR+ and AR− prostate cancer cells to IR. Second, exogenous expression of AR only partially rescues the impairment of IR-induced DSB repair by PRMT5 knockdown. Third, PRMT5 knockdown increases IR-induced DSB in AR− DU145 cells and several other cancer cell lines and normal cells. Fourth, PRMT5 expression correlates positively with the expression of its target genes in multiple human cancer tissues. During preparation of this project, Braun et al. reported that PRMT5 post-translationally regulates the splicing out of detained-introns (DI)s of genes to modulate gene expression. However, analysis of their data showed that the majority of DEGs we identified either do not contain DIs or DI splicing was not affected by targeting PRMT5. In addition, Clarke et al. reported that PRMT5 participates in the DSB repair choice process and promotes HR through methylation of RUVBL1. It is therefore likely that PRMT5 regulates repair of IR-induced DSB via multiple mechanisms. As PRMT5 is overexpressed in many human cancers and its overexpression correlates with poor prognosis, our findings suggest that increased DSB repair by PRMT5 overexpression in these human cancers may confer survival advantages particularly following DNA damaging treatment. Because targeting DSB repair has been proven to be a valid therapeutic approach for cancer treatment, our findings here also suggest that PRMT5 targeting may be explored as a monotherapy or in combination therapy with RT or chemotherapy for cancer treatment.

OBJECTIVES/GOALS: There is a high burden of lung cancer in persons living with HIV (PLWH). The role that HIV status, by levels of immune function and viral load, has on survival from lung cancer is not fully understood. The study’s objectives were to assess 1) the association of HIV with survival in non-small cell lung cancer (NSCLC) and 2) prognostic factors in PLWH with NSCLC. METHODS/STUDY POPULATION: Participants were from a cohort of lung cancer patients diagnosed between 2004-2017 in the Bronx, NY, with vital status ascertainment at least annually. We compared survival from NSCLC diagnosis between HIV-negative patients (HIV-, N = 2881) and PLWH (N = 88), using Cox regression, accounting for clinical and sociodemographic factors including smoking status. In three separate comparisons to HIV-, PLWH were dichotomized by CD4 count (<200 vs. ≥200 cells/μL), CD4/CD8 ratio (median, <0.43 vs. ≥0.43) and HIV viral load (VL) suppression (<75 vs. ≥75 copies/mL). In PLWH only, we assessed the relationships of CD4 count, CD4/CD8 ratio, and VL at diagnosis with survival adjusting for age, sex, and cancer stage. CD4 count and CD4/CD8 ratio were also examined as time-varying variables using a counting process approach. RESULTS/ANTICIPATED RESULTS: PLWH were younger (median 56 years, IQR 51-52 vs. 68, IQR 60-76) and more likely to be current smokers (58% vs. 37%) at diagnosis than HIV- patients. Median survival was lower in PLWH [1.1 years, 95% confidence interval (95%CI): 0.6-1.3] than in HIV- [1.6 (1.5-1.7)]. Survival comparing PLWH with higher CD4/CD8 to HIV- was similar [hazard ratio (HR), 95%CI: 0.63 (0.37-1.07)], but those with lower CD4/CD8 experienced worse survival (HR = 1.74, 95%CI: 1.07-3.89). Among PLWH, having a CD4 count < 200 cells/μL was associated with over twice the risk of death compared to those with CD4 ≥ 200 cells/μL (HR = 2.37, 95%CI: 1.14-4.92). VL and CD4/CD8 ratio were not associated with survival. Lower time-updated CD4 count was also associated with worse survival (HR = 2.19 for CD4 <200 vs. >200 cells/μL, 95%CI: 1.16-4.13). DISCUSSION/SIGNIFICANCE OF IMPACT: Among persons with NSCLC, CD4/CD8 ratio nearest diagnosis was shown to distinguish mortality risk in PLWH compared with HIV- patients. In addition, PLWH with low CD4 had worse prognosis than PLWH who had higher CD4 counts. These results suggest HIV immune status to be an essential component influencing survival in lung cancer.

OBJECTIVES/GOALS: Childhood Sjögren’s disease (cSD) is a rare autoimmune disease. Despite the profound impact on children and their families, pediatric-specific clinical trials to inform therapeutic strategies in cSD are lacking. In 2022 we participated in the Trial Innovation Network (TIN) Design Lab with the purpose of designing a series of N-of-1 trials for cSD. METHODS/STUDY POPULATION: New medications have the potential to be safe/effective treatments for cSD but must be evaluated in randomized trials. To overcome limitations of traditional parallel-group designs given the rarity of cSD, we developed an N-of-1 trial approach. Our proposal was selected by the Tufts TIN Design Lab. The Design Lab multi-stakeholder process involved parents of and patients with cSD, pediatric and adult rheumatologists, and experts in clinical trial design and outcomes. We engaged all stakeholders in protocol development to maximize the impact of the proposed approach on clinical care, ensure a successful recruitment plan, and inform the choice of endpoints as there are no widely accepted cSD outcome measures to determine treatment efficacy. RESULTS/ANTICIPATED RESULTS: Using the Design Lab methodology, we clarified the N-of-1 study goals and engaged in an iterative process to develop a “briefing book” that ensured a sound premise for our study. We reviewed and accumulated published literature to support our focus on mucosal/glandular manifestations, identified potential interventions to be used in the N-of-1 trials, and enumerated possible outcomes, including outcomes important to patient/parents. This work culminated in a full-day Design Lab event that included multiple stakeholders who provided expertise from different perspectives on the full drug development pathway. Study design feedback focused on three specific areas. 1) Inclusion and exclusion criteria; 2) Identification of outcome measures; 3) Treatment and washout periods. DISCUSSION/SIGNIFICANCE: To address the critical need and move treatment of cSD forward, we are designing a prototype N-of-1 trial in children with rheumatic disease. We will continue to engage stakeholders by using a series of Delphi surveys and an in-person meeting to create composite outcome measures to test cSD therapies in personalized trials.

OBJECTIVES/GOALS: modonko1@jhmi.edu METHODS/STUDY POPULATION: Michelle Odonkor RESULTS/ANTICIPATED RESULTS: I accept DISCUSSION/SIGNIFICANCE: No, I do not want my poster published JCTS.

OBJECTIVES/GOALS: To describe and evaluate an innovative university-community vaccination and food access model for minority, immigrant, and underserved individuals experiencing food insecurity during a global pandemic. METHODS/STUDY POPULATION: The Purdue University Center for Health Equity and Innovation (CHEqI) partnered with the two largest food banks in the Midwest and Walgreens to offer free COVID-19 and Flu vaccinations alongside food distribution. Goals included addressing food insecurity, increasing vaccine access, and decreasing vaccine hesitancy. CHEqI acquired funding, recruited volunteers and interpreters, assessed interest and addressed vaccine hesitancy. Food bank/pantry partners distributed food and provided access to clientele and marketing assistance. Walgreens procured, administered, and documented vaccinations. The Model accommodated drive-through and indoor processes. Unidentifiable observational and self-report data were collected. Descriptive statistics were computed to characterize program outcomes. RESULTS/ANTICIPATED RESULTS: A total of 11 vaccination events occurred between June and October 2021 at three food bank/pantry locations. Of these 11 events, nine (82%) were drive-through and two (18%) took place indoors, eight (72%) offered COVID-19 vaccinations only, and three (27%) offered both COVID-19 and Flu vaccinations. Food was distributed to a total of 5,108 families and 416 vaccines (314 COVID, 102 Flu) were administered. Of the 396 individuals who received at least one vaccine, 20 (5%) received both a COVID and Flu vaccine. Of the 386 individuals who received at least one vaccine and reported their sex, 194 (50%) identified as female and the average age of those who received at least one vaccine was 45 years old. Of those who reported race (N = 228) or ethnicity (N = 253), 43% identified as Black or African American and 53% identified as LatinX. DISCUSSION/SIGNIFICANCE: Findings offer an innovative vaccination and food access model for diverse individuals experiencing food insecurity during a global pandemic. By drawing on cost effective, accessible, and culturally contextualized practices to optimize the reach and quality of vaccination services we can improve access barriers and mitigate health disparities.

OBJECTIVES/GOALS: The purpose of this study is to evaluate average pain scores in patients with Hidradenitis Suppurativa (HS) and determine if racial disparities influence HS-related pain management. METHODS/STUDY POPULATION: We surveyed 3,140 adult patients about demographics, HS, pain levels, medical history, and health-related quality of life. Among respondents (N=162), pain scores (0-10) were grouped as high (8+, N=54) and low (<7, N=108). We used logistic regression to calculate adjusted odds ratios (aOR) and 95% confidence intervals (95% CI). RESULTS/ANTICIPATED RESULTS: Respondents were female (88.9%), Black race (59.4%), with a median age of 36 (IQR 25-42) and a mean pain score of 7 (IQR 5-8). Half (48.2%) of respondents disagreed or strongly disagreed with, “I am satisfied with how my pain related to HS is being managed by my doctors.” Independent risk factors for high pain included Black race (aOR=7.95, 95% CI 3.11-20.26), history of prescribed pain medications (aOR=2.37, 95% CI 1.06-5.30), and former/current tobacco use (aOR=4.12, 95% CI 1.76-9.64). DISCUSSION/SIGNIFICANCE: The preliminary data support the hypothesis that race influences HS-related pain. In addition, less education, a history of prescription pain medication use, and tobacco use are all independent risk factors contributing to higher pain scores in patients with HS. Further analysis will be conducted to evaluate racial influences on pain management.

OBJECTIVES/GOALS: This theory analysis aims to evaluate a middle-range framework, the HSDF1, in the context of sexual assault stigma incorporating the myriad levels within within culture and society through which stigma can occur and be reinforced. METHODS/STUDY POPULATION: Databases: PubMed, CINAHL, EMBASE, Google Scholar, Organization websites, Citation searchesn = 32Mark Risjord’s “Middle-Range Theories as Models: New Criterion for Analysis and Evaluation” (2019) RESULTS/ANTICIPATED RESULTS: The innovative approach of the HSDF guides understanding of sexual assault stigma in a holistic way, incorporating individual and institutional stratum of the phenomenon. Understanding through integration of this theoretical framework alongside current knowledge may more succinctly inform trauma-informed care for survivors, policy, and cultural awareness for nurses, healthcare providers, police, social workers, and myriad others with whom survivors interact. DISCUSSION/SIGNIFICANCE: Applying the HSDF framework to sexual assault stigma could help break down barriers and raise survivors out of stigmatization, affecting population health through reduced negative health sequelae experienced by survivors.

ABSTRACT IMPACT: This work investigates C. difficile strains in soil as a potential exposure for gut colonization and community-acquired infection of C. difficile. OBJECTIVES/GOALS: Identifying environmental sources of C. difficile can inform how non-hospital reservoirs can potentially contribute to C. difficile exposure and subsequent gastrointestinal colonization. The objective of the study was to identify C. difficile and toxin genes across various soil sources. METHODS/STUDY POPULATION: This was a cross-sectional study utilizing soil samples obtained throughout Texas, USA. All samples were collected between August and November of 2019 and 2020. Samples were taken from human and animal high contact areas, such as recreational parks. Samples were stored at -80oC until processing. DNA extractions were performed using the DNeasy Powersoil Pro Kit (Qiagen) per manufacturer’s instructions. Real-time PCR was also performed on extracted DNA using the Microbial DNA qPCR Multi-Assay Kit for Clostridium difficile Pathogenicity (Qiagen) for the identification of C. difficile, toxin A (TcdA), and toxin B (TcdB) genes. RESULTS/ANTICIPATED RESULTS: A total of 137 soil samples including dry dirt, sand, and wet soil near water sources were collected and processed for the presence of C. difficile. These included samples from parks and trails (42.3%), water sources (36.5%), and other public spaces (21.2%). C. difficile was identified in 59 (43.1%) soil samples: 6 (4.4%) with Toxin A and 2 (1.5%) with toxin B production. C. difficile was most prevalent among samples taken from parks and trails (50.0%), followed by other public spaces (48.3%), and water sources (32.0%). The median (IQR) Cq value for the C. difficile gene was 39.24 (33.45-40.47) among samples that tested positive. DISCUSSION/SIGNIFICANCE OF FINDINGS: We identified a high prevalence of Clostridioides difficile in soil samples, though toxin gene detection prevalence was low. Future studies will analyze other sources, including water and varying surface samples to obtain a comprehensive view of C. difficile in the environment.

OBJECTIVES/SPECIFIC AIMS: In the United States, it is estimated that approximately half of all pregnancies are unintended. This study examines the prevalence of unintended pregnancy in a cohort of cancer survivors and identifies factors associated with unintended pregnancy after cancer. METHODS/STUDY POPULATION: The FUCHSIA Women’s Study is a population-based study of female cancer survivors at a reproductive age of 22–45 years. Cancer survivors diagnosed between the ages of 20 and 35 years and at least 2 years postdiagnosis were recruited in collaboration with the Georgia Cancer Registry. Participants were interviewed about their reproductive histories. The prediagnosis analysis included all women who completed the interview; the postdiagnosis analysis excluded those who had a hysterectomy, bilateral oophorectomy, or tubal ligation by cancer diagnosis. RESULTS/ANTICIPATED RESULTS: Of the 1282 survivors interviewed, 57.5% reported at least 1 pregnancy before cancer diagnosis; of which, 44.5% were unintended. Of the 1088 survivors included in the postdiagnosis analysis, 36.9% reported a post-cancer pregnancy. Among those who had a pregnancy after cancer diagnosis, 38.6% reported at least 1 pregnancy was unintended. Of the 80 breast cancer survivors who had a pregnancy after diagnosis, 52.5% of them were unintended. Predictors of unintended pregnancy in cancer survivors included being younger than 30 years at diagnosis [odds ratio (OR) 2.1; 95% confidence interval (CI) 1.4, 2.9], identifying as Black (OR 1.6, 95% CI 1.1, 2.3, comparison: White), and having resumption of menses after cancer treatment (OR 8.1, 95% CI 2.0, 33.0). Compared with being <4 years from cancer diagnosis, those who were farther from diagnosis at the time of the interview also had increased odds of unintended pregnancy (4–7 years: OR 1.5, 95% CI 0.9, 2.7; 8–10 years: OR 1.3, 95% CI 0.7, 2.4; >10 years: OR 2.7, 95% CI 1.6, 4.7). DISCUSSION/SIGNIFICANCE OF IMPACT: Despite being at higher risk of infertility, cancer survivors may still be at considerable risk of unintended pregnancy. Women with certain types of cancer that are more likely to be hormone responsive, such as some types of breast cancer, may be hesitant to use hormonal birth control and thus be at higher risk of unintended pregnancy. Counseling for cancer survivors should include a discussion of the risk of unintended pregnancy and contraceptive options.

OBJECTIVES/SPECIFIC AIMS: Cardiovascular disease (CVD) is the leading cause of death among US adults and its prevalence is increasing, despite efforts to identify, and address risk factors. Post-traumatic stress disorder (PTSD) has been identified as a potential risk factor for CVD, though the results to date have focused on male veterans with combat-related PTSD. To our knowledge, there are no prospective analyses/reports among older community-dwelling adults following Hurricane Katrina. The purpose of this study was to explore the link between PTSD associated with Hurricane Katrina and incident CVD among elderly adults using data from the Cohort Study of Medication Adherence among Older Adults (CoSMO). METHODS/STUDY POPULATION: PTSD associated with Katrina and incident CVD events were assessed among 2075 hypertensive participants age ≥65 who were enrolled in a managed care organization in southeastern Louisiana. Baseline surveys were conducted between August 2006 and September 2007. Baseline surveys were conducted between August 2006 and September 2007. PTSD was assessed using the civilian PTSD CheckList (PCL-17) and 2 cut-off points, ≥37 and ≥44, for primary and secondary analyses, respectively. Participants were followed through February 2011 for a composite CVD outcome of MI, stroke, CHF, or CVD death. Multivariable logistic regression was performed with 13 covariates identified in bivariate analysis: age, sex, race, marital status, education, hypertension knowledge, comorbidities, number of antihypertensive medication classes, dissatisfaction with healthcare, reduced medications due to cost, number of visits to healthcare provider in last year, depression, and coping. RESULTS/ANTICIPATED RESULTS: Participants were 59.8% female and 30.4% black, with a mean age of 75 years. The prevalence of PTSD using the primary and secondary cut points was 6.1% and 4.2%, respectively. In total, 240 (11.5%) participants had a CVD event during a median 3.8 year follow-up. After multivariable adjustment, the odds ratios and 95% confidence intervals (CI) for CVD event for the primary and secondary analyses were 1.90 (95% CI: 1.17, 3.09) and 3.74 (95% CI: 2.05, 6.81), respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: PTSD was associated with an increased risk of incident CVD events among elderly adults. This finding from a prospective cohort study supports earlier reports suggesting PTSD is an independent risk factor for CVD. To our knowledge, this association has not been previously reported among a cohort of elderly community-dwelling adults. This study included hypertensive, elderly, insured participants living in southeastern Louisiana following Hurricane Katrina and may not be generalizable to all people with PTSD. Strengths of this study include its longitudinal design, the identification of incident CVD, the diversity of the study population with respect to gender, race and CV risk, and reduced confounding due to access to care and insurance status. Future research is needed to confirm this finding in other populations and to assess if efforts to minimize the impact of PTSD following disasters reduce CVD risk and premature CVD events among older adults.

Objectives/Goals: Our goal is to enhance communication and documentation in collaborative biostatistics by refining data management and metadata processes. We aim to capture critical data collection and generation information, improve transparency and reproducibility, and foster stronger researcher partnerships for more effective collaborations. Methods/Study Population: Traditional statistical analysis plans (SAP) often miss essential contextual knowledge from collaborators, leading to gaps that hinder reproducibility and limit future data use. Biostatistics teams at the University of Kentucky have updated their strategies to better capture important details about data origins and collection processes. By focusing on clear, comprehensive documentation early in the research process, we aim to preserve foundational data insights and improve collaboration efficiency. Our Biostatistics, Epidemiology, and Research Design (BERD) team has established best practices for addressing data management structures with collaborators across medical and healthcare fields – covering all project stages, from initial data collection to metadata creation and dataset finalization. Results/Anticipated Results: We will detail the processes used to improve data management structures and the observed results of these processes. For example, initiating deeper discussions about data origins and collection processes as early as possible in the collaboration has resulted in a more comprehensive project narrative that lays the foundation for effective collaboration. By engaging with project leaders early in the process, we can confirm that critical details about how data were collected and processed are documented, improving both the transparency and reproducibility of research findings. Streamlining the processes of capturing this information makes it more accessible and useful for those with limited statistical backgrounds, which is particularly relevant for faculty and staff in BERD communities and Clinical and Translational Science Awards Programs. Discussion/Significance of Impact: Nuanced data documentation structures are crucial for transforming raw data into meaningful, reusable datasets. Our initiatives promote clear communication, enhanced efficiency, and streamlined workflows. Translational science researchers can benefit from improving data management and metadata to boost long-term collaborative success.

OBJECTIVES/GOALS: To increase the diversity of the health sciences research workforce, students from a variety of backgrounds must have the opportunity to participate in hands-on research experiences that highlight translating science to treating human disease. We developed a mentored translational research program for students from VCU and central Virginia HBCUs. METHODS/STUDY POPULATION: The Wright Regional Center for Clinical and Translational Science collaborated with the existing VCU Honors’ Summer Undergraduate Research Program (HSURP) to expand their summer research experience to URM students from our partner HBCUs. For 10 weeks, students worked with faculty mentors to learn research techniques and engage in research projects. Students also participated in career development sessions like developing a CV and choosing graduate programs, and at the end of HSURP, they shared formal presentations of their research with peers and mentors. HSURP students were provided housing and a stipend, and mentors were provided a stipend. A post-program assessment gathered feedback on research and personal skills gained, the program’s influence on their career goals, and overall experiences with HSURP. RESULTS/ANTICIPATED RESULTS: Nine students, 7 from VCU and 2 from Virginia State University participated in HSURP. Students were rising sophomores, juniors, and seniors, and 5 had previous research experience. Students worked on projects ranging from basic to social behavioral, community-placed research. All students rated the program as good or excellent. Post-program assessments showed all students believed they had a better understanding of ethical responsibilities of researchers, relevance of community-engaged and clinical/translational research, and interpreting journal articles after participating in the program. Four students reported they plan to continue working on their research projects during the academic year, and all students strongly agreed or agreed that HSURP prepared them for graduate or professional schools. DISCUSSION/SIGNIFICANCE: A program that combines hands-on research training and career development opportunities provides a robust research foundation for URM students, which can increase their participation in the translational science workforce. Future program development will include preprogram training modules to better prepare students for research experiences.

ABSTRACT IMPACT: By demonstrating the feasibility of a multimodal, interdisciplinary intervention for prediabetes, the current project aims to provide a template for the prevention of diabetes and associate comorbid conditions. OBJECTIVES/GOALS: To determine if a multi-modal, interdisciplinary intervention delivered to a group of prediabetic patients will result in reduced rates of diabetes progression. This project is a retrospective evaluation that will exam the feasibility and possibly efficacy of this intervention. METHODS/STUDY POPULATION: We will enroll 50 participants for the clinic, aged 21-60 inclusive. Patients will have a Body Mass Index >27kg/m2 with a diagnosis of prediabetes. Patients must be non-pregnant, using approved contraception, and agree to not become pregnant for 1 year after enrollment. After enrollment, the initial treatment period is for 1 year and includes a 12 week low calorie diet plan, a 6-month intensive behavioral and lifestyle modification plan followed by a 6 month behavior reinforcement extension. Weight management medications may be used if appropriate for the patient from a clinical perspective during the 6-month intensive behavioral/lifestyle modification. RESULTS/ANTICIPATED RESULTS: It is anticipated that there will be decreased weight with a mean weight loss goal of approximately >10%. Furthermore, it is expect that there will be improvement of other markers of metabolic disease. These include improvement of lipid values (LDL-C, HDL-C, Triglycerides, Total Cholesterol) as well as blood pressure with expected blood pressures of below 130/80 in greater than 50% of participants. Finally, It is expected that 50% or greater participants will have improvement of glycemic control. It is anticipated that greater than 50% of participants will have improvement of glycemic control and achieve normoglycemia. These values will be determined based upon fasting glucose or A1c. DISCUSSION/SIGNIFICANCE OF FINDINGS: The significance of this intervention is enormous. By demonstrating feasibility in this trial, we can work toward both assessing efficacy and possibly dissemination of this model program. If these interventions provide durable changes at scale, this could help slow the epidemic of obesity and obesity related comorbid conditions.

Objectives/Goals: In mice, it has been shown that loss of Cib2 (calcium and integrin-binding protein 2) results in progressive retinal disease that recapitulates many characteristics of age-related macular degeneration (AMD). This study aims to characterize transcriptional changes in the retinal pigment epithelium (RPE) that underlie this disease process. Methods/Study Population: RPE tissue samples, pooled from 2–3 mice for each biological group, were collected from Cib2-KO and wildtype (WT) mice at two (young) and eight (aged) months of age. Bulk mRNA sequencing was performed using the Illumina HiSeq 4000. Reads were aligned to the UCSC mouse reference genome and quantified using HTSeq. Significant differentially expressed genes (DEGs) between mouse genotype and age groups were assessed using DESeq. CLICK unsupervised clustering followed by gene ontology analysis was performed to identify cellular processes and molecular pathways affected by loss of Cib2 as well as age. Results/Anticipated Results: CLICK analysis revealed several functional pathways that are differentially expressed between sample groups. For example, in both young and aged mice, pathways upregulated in Cib2-KO samples included calcium signaling, RhoA signaling, and integrin signaling. Uniquely downregulated DEGs in young Cib2-KO animals were related to complement and coagulation cascades, LXR/RXR activation (related to lipid synthesis and transport), and phagosomes. Aged Cib2-KO mice displayed the most significant downregulation of genes in the phototransduction pathway, indicating temporal changes in functional pathways that correlate with disease progression. Next steps in analysis include investigating patterns in RPE- and AMD-signature gene sets that may identify molecular pathways more specific to human disease. Discussion/Significance of Impact: Many current studies investigate the role of complement activation, vesicle trafficking, and ion transport as top contributors to AMD development. We identified DEGs paralleling many of these molecular pathways in Cib2-KO mice, highlighting their potential as a model to study age-related RPE pathologies and evaluate therapeutic interventions.

OBJECTIVES/GOALS: Studying IPF associated health care utilization (HRU) in real world settings, provides the opportunity to produce generalizable results that can directly inform models of care delivery. The objective of this study was to examine real-world differences in the natural history of annual HRU and treatment trends associated with IPF in a large, community-based population of patients with IPF, compared to matched non-IPF controls. METHODS/STUDY POPULATION: Cases of IPF were identified using case validated algorithms in the Kaiser Permanente Northern California adult population from 2000 to 2014. Each case was matched to at least one and no more than five non-IPF controls by age, sex, race/ethnicity and length of enrollment. The date of the first occurrence of the IPF-specific diagnostic code was considered the index date for cases and matched controls. Comorbidity burden and HRU was assessed in the five years pre- and post-index date, including hospitalizations, outpatient visits, use of diagnostic and monitoring studies and medications. Poisson generalized estimating equations models with robust standard errors were used to estimate adjusted case-control differences in HRU, accounting for clustering within matched sets. RESULTS/ANTICIPATED RESULTS: 691 patients were identified with incident IPF during the study period and matched to 3,452 control subjects. Demographics were well balanced between cases and controls due to matching. Patients with IPF had a higher burden of all selected co-morbidities and higher HRU compared to controls. In the pre-index period, IPF members had significantly higher rates of all diagnostic procedures (p < 0.001) and health care visits, with the exception of skilled nursing facility care (p < 0.001). The greatest relative difference was observed with use of Chest CT (RR = 245.94, 95% CI 117.04, 516.82). In the post-index period compared to controls, patients with IPF had higher rates of serial testing (p < 0.001) and inpatient and outpatient care including, all-cause hospitalization (1.55), emergency room visits (1.19), outpatient visits (1.18), and skilled nursing facility stay (1.35). DISCUSSION/SIGNIFICANCE OF IMPACT: Patients with idiopathic pulmonary fibrosis experience increased co-morbidity and healthcare resource utilization compared to controls. This increased burden extends beyond the index-period as previously documented and is true for a large real-world cohort. CONFLICT OF INTEREST DESCRIPTION: NA

OBJECTIVES/SPECIFIC AIMS: The study aims to further investigate how cyclophilin D (CypD), the key mPTP opening regulator, affects BMSCs fate and to determine potential regulatory mechanisms involved in CypD regulation during osteogenesis. METHODS/STUDY POPULATION: We evaluated CypD mRNA expression in mouse BMSCs and in osteogenic-like (OL) cells during the course of OB differentiation. CypD protein level was also probed. Moreover, BMSCs had their mPTP activity recorded during osteoinduction. We further analyzed the effect of CypD genetic deletion on osteogenesis in vitro and in vivo. For our in vivo model, we performed the ectopic bone formation assay to asses differences in ossicle formation when CypD KO BMSCs were transplanted compared to wild type littermate BMSCs. In our in vitro model, we transfected OL cells with either CypD gain of function or CypD loss of function vector and measured their osteogenic differentiation potential. Additionally, we treated BMSCs with CypD inhibitor and compare to non-treated BMSCs for mineralization level. To determine potential regulatory mechanisms involved in CypD regulation, we analyzed the CypD gene (Ppif) promoter for potential transcription factor (TF) binding sites and found multiple Smad-binding elements within this promoter. Smads (Smad1, 5, 8) are TFs downstream from Bone Morphogenic Protein (BMP) signaling pathway that transmit cell differentiation signaling, and exert either activating or inhibitory effects on a variety of genes. We also transfect OL cells with Smad1 vector and analyzed for CypD mRNA levels. RESULTS/ANTICIPATED RESULTS: - Our data showed that CypD mRNA levels decreased in both primary cells and OL cells at day 7 and day 14 in osteogenic media. - Osteogenic induction also decreased mPTP activity. - In vivo ectopic bone formation assay showed increased ossicle fo DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that downregulation of CypD increases OB differentiation due to improved OxPhos activity led by mPTP closure. Our results corroborate reports of CypD downregulation and mPTP closure during neuronal differentiation in developing rat brains as well as in cardiomyocyte differentiation in developing mouse hearts. Our studies also suggest a yet unknown mechanism linking differentiation signaling with mitochondrial function – BMP/Smad mediated downregulation of CypD transcription. As initially mentioned, in a previous study, our lab showed that CypD KO mice present higher mitochondrial function and osteogenicity in aged BMSCs and less osteoporosis burden. Taken together, these results suggest that CypD can be a potential target to prevent bone loss in aging.