Interprofessional healthcare team function is critical to the effective delivery of patient care. Team members must possess teamwork competencies, as team function impacts patient, staff, team, and healthcare organizational outcomes. There is evidence that team training is beneficial; however, consensus on the optimal training content, methods, and evaluation is lacking. This manuscript will focus on training content. Team science and training research indicates that an effective team training program must be founded upon teamwork competencies. The Team FIRST framework asserts there are 10 teamwork competencies essential for healthcare providers: recognizing criticality of teamwork, creating a psychologically safe environment, structured communication, closed-loop communication, asking clarifying questions, sharing unique information, optimizing team mental models, mutual trust, mutual performance monitoring, and reflection/debriefing. The Team FIRST framework was conceptualized to instill these evidence-based teamwork competencies in healthcare professionals to improve interprofessional collaboration. This framework is founded in validated team science research and serves future efforts to develop and pilot educational strategies that educate healthcare workers on these competencies.

OBJECTIVES/GOALS: 48,000,000 people in the U.S. have hearing loss, negatively impacting quality of life and work. Unveiling axon guidance for auditory type II spiral ganglia neurons (SGNs) will aid development of new therapies. I study the role of Eph/Ephrin and planar cell polarity (PCP) signaling during type II SGN turning and outer hair cell (OHC) innervation. METHODS/STUDY POPULATION: This quantitative study was conducted on Efna3 and Vangl2 null mice possessing Neurog1CreERT2 and R26RtdTomato mutations. Spontaneous Cre activity within the Neurogenin1CreERT2 line causes recombination and expression of fluorescent Rosa26 Reporter (R26R)tdTomatoin a restricted number of SGNs, including type IIs. Together, these lines permit SGN sparse labeling. Immunostaining and confocal imaging were used to analyze dsRed in Efna3 and Vangl2 mice and quantify type II SGN turning. In combination, Imaris 3D renderings were used to quantify type II SGN turning, branching, navigation features and temporal effects of EPHRIN-A3-Fc on type IIs via cochlear cultures (a gain-of-function manipulation). For both sexes, 5-6 cochleae per genotype were analyzed and compared by t-test to wildtype (WT) controls. RESULTS/ANTICIPATED RESULTS: Efna3 nulls showed a small rise in type II SGNs incorrectly turning toward the apex at an error rate of 16.0% compared to WTs (n=6; p=0.05). P0 Efna3 nulls had reduced branch number compared to WTs, 4.1 and 7.2, respectively (n=129; p=<0.0001), suggesting EPHRIN-A3 acts as a positive growth cue. In cochlear cultures, EPHRIN-A3-Fc led to type II SGN collapse at E15.5, indicating a repulsive function. However, at P0, EPHRIN-A3-Fc treatment led to type II SGNs with elevated branch numbers compared to Control-Fc treatment: 18.1 and 11.4, respectively (n=116; p=<0.0001). This indicates a positive growth function. At E16.5, EPHRIN-A3 protein immunoreactivity on Deiters’ and pillar cells appears reduced in Vangl2 nulls compared to WT cochleae, suggesting that EPHRIN-A3 acts downstream of PCP signaling. DISCUSSION/SIGNIFICANCE: Results suggest that Eph/Ephrin signaling acts downstream of PCP signaling to mediate type II SGN guidance and EPHRIN-A3 switches its mode of activation. The clinical implications of these findings are that therapeutics targeting EPHRIN-A3 and/or VANGL2 in their given pathways could stimulate new OHC innervation following auditory damage.

OBJECTIVES/SPECIFIC AIMS: In the present study, we examined the functional consequences of 3α-PVA on OATP1B1-mediated PVA transport. To elucidate this, we determined the effect of SLCO1B1 genotype on PVA transport and the role of 3αPVA as a competitive inhibitor of OATP1B1, which could serve as another covariate that disrupts the systemic and hepatic exposure of pravastatin in children and adults. METHODS/STUDY POPULATION: Site directed mutagenesis was performed to generate SLCO1B1 genotypes of interest (*1a, *1b, *5, *15). Human embryonic kidney (HEK293) cells were grown and plated at 200 000 cells per well in 24-well plates. Twenty-four hours later the cells were transfected with the aforementioned plasmids. Forty-eight hours later cell-based transport was performed with radiolabelled [3H]-pravastatin sodium salt. Non-radioactive pravastatin sodium salt and 3’α-iso-pravastatin sodium salt was used for PVA transport and 3αPVA studies, respectively. Cells were washed 3 times with warm uptake buffer, incubated for 1 minute with uptake solutions containing PVA and 3αPVA at varying concentrations. Transport was terminated by four 1-ml washes with ice-cold uptake buffer. Cells were lysed with 300 µl 1% Triton X-100 in PBS at room temperature for 30 minutes prior to analysis. Radioactivity was measured in a MicroBeta2 liquid scintillation counter. The remaining cell lysates were transferred to 96-well plates to determine total protein concentration using the bicinchonic acid protein assay. All transport measurements were corrected by the total protein concentration. All experiments were performed 3 to 4 times independently with 2-3 determinations. Data were analyzed for significant differences amongst genotype groups using ANOVA followed by Tukey’s multiple comparisons test. IC50 and kinetic parameters were calculated using non-linear regression analysis. RESULTS/ANTICIPATED RESULTS: Pravastatin transport in SLCO1B1 variants (*5, *15) was significantly decreased compared to the reference genotype *1a and *1b (Km [µM]: *1a 18.2 ± 0.9; *1b 17.9 ± 3.3; *5 34.2 ± 9.7; *15 34.1 ± 6.1; p≤0.05; Vmax [pmol/mg/min]: *1a 104.9 ± 13.1; *1b 93.7 ± 16.7; *5 44.8 ± 15.9; *15 62.3 ± 22.5; p≤0.05). *1a and *1b were not significantly different with respect to pravastatin transport. Intrinsic clearance was diminished nearly 4 to 5-fold in SLCO1B1 variants compared to reference genotypes (Vmax/Km [µl/min/mg]: *1a 5.8 ± 0.8; *1b 5.7 ± 1.9; *5 1.3 ± 0.2; *15 1.8 ± 0.3; p≤0.01). Pravastatin transport was inhibited by 3αPVA for all genotypes. However, there was more pronounced inhibition in the SLCO1B1 variant genotypes compared to reference genotypes (IC50 [µM]: *1a 15.9 ± 1.9; *1b 18.6 ± 5.7; *5 3.9 ± 2.0; *15 4.4 ± 0.8; p≤0.01; Ki: *1a 15.0 ± 1.8; *1b 17.5 ± 5.4; *5 3.8 ± 2.9; *15 4.3 ± 0.8; p≤0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: In vitro PVA transport is altered according to SLCO1B1 genotype, consistent with previous in vitro and human experience. Our data suggest that the significantly different maximal transport velocity (Vmax) in variant versus non-variant genotypes is consistent with decreased membrane expression of OATP1B1 with the variant c.521T>C allele. However, in contrast to data involving typical model substrates (e.g. estrone-3-sulfate), the PVA binding affinity (Km) was significantly different between variant and non-variant genotypes, consistent with altered binding of the substrate to OATP1B1. Collectively, we conclude that decreased OATP1B1 expression and function in variant genotypes influence altered transport for PVA. Finally, the functional consequences of 3αPVA formation on PVA transport was confirmed in our study. Mechanistically, we confirmed our observation in humans that 3αPVA inhibits OATP1B1 transport. However, this effect is more pronounced in variant genotypes as shown by lower IC50 values compared to the reference genotypes. This highlights another source of variation that must be taken into consideration when trying to optimize the pravastatin dose-exposure relationship in humans.

OBJECTIVES/GOALS: CTRUs support clinical research. RUN is a Learning Research System that is created to enhance CTSA and non-CTSA research units capacity through implementing, assessing, and disseminating discoveries in methods, approaches, education, and training in clinical and translational science. METHODS/STUDY POPULATION: The RUN association began in July 2018 with eight universities. The association has grown to 44 hospitals, research, and academic institutions (including 36 CTSA institutions). A RUN Discussion Forum has been approved by the National Center for Advancing Science (NCATS) and utilized by RUN. The Discussion Forums are created with the goal of advancing CTSA Program objectives in high priority areas of clinical and translational science. RUN actively engages members through in depth scheduled monthly meeting discussions with various relevant topics regarding the development and evaluation of clinical trials metrics, benchmarks, and scholarly publication and presentation activities. RESULTS/ANTICIPATED RESULTS: Topics covered in RUN monthly meetings include research units general budget guidelines, staff recruitment and retainment strategies, EPIC use in scheduling CRU research visits, and PPE for investigational drugs in context of USP800 requirements. RUN members vary in geographic location, type of clinical research (outpatient vs inpatient), resources, and research subject volume. They are engaged in online discussion and learning opportunities to improve translational science practices. A recent article titled “Impact of COVID-19 on Clinical Research Units (CRUs)” in JCTS is an example of best practices learned by RUN members and shared with the broader research community. DISCUSSION/SIGNIFICANCE: RUN as a Learning Research System enhances clinical and translational research unit capacity and efficiency, encouraging collaboration to contribute with improving public health. This network is aligned with the CTSAs mission of developing innovative solutions to improve translational science.

OBJECTIVES/GOALS: The study aims to explore the role of microglial behavior in cognitive impairment associated with vascular dementia (VaD) and long COVID. Using immunohistochemistry (IHC) and quantitative PCR (qPCR), we will assess the expression of Iba-1, a microglial activation marker, in subjects with VaD and SARS-CoV-2 infection. METHODS/STUDY POPULATION: Out of 48 female C57BL/6 mice, 24 had surgical intervention in the form of bilateral carotid artery stenosis (BCAS) for experimental induction of vascular dementia. After 2 weeks, 12 BCAS and 12 non-BCAS were infected with 1E4 PFU of mouse-adapted 10 (MA10) strain of SARS-CoV-2. 2 weeks post-infection, 4 weeks post-operatively, all animals were euthanized and tissues were processed for cDNA and histology. Immunofluorescence and RT-qPCR used to quantify microglia via Iba-1, BBB integrity via claudin-5 as well as occludin, GFAP, and integrin a5. RESULTS/ANTICIPATED RESULTS: We anticipate observing distinct patterns of microglial behavior in subjects with vascular dementia (VaD) and those with long COVID. Through immunohistochemistry (IHC), we expect to see increased Iba-1 expression, indicative of microglial activation. Quantitative PCR (qPCR) will likely corroborate these findings, showing elevated levels of Iba-1 mRNA. Lastly, we anticipate that the data will reveal interactions between microglia and the blood-brain barrier (BBB). These interactions could provide insights into how microglial behavior influences BBB integrity and, consequently, cognitive function in VaD and long COVID. DISCUSSION/SIGNIFICANCE: This study aims to clarify the role of microglia in cognitive decline linked to vascular dementia and long COVID. By categorizing patients based on microglial activation, we can better tailor treatments. The findings could lead to targeted therapies that address cognitive impairment in these conditions.

OBJECTIVES/GOALS: Large-scale clinical proteomic studies of cancer tissues often entail complex workflows and are resource-intensive. In this study we analyzed ovarian tumors using an emerging, high-throughput proteomic technology termed SWATH. We compared SWATH with the more widely used iTRAQ workflow based on robustness, complexity, ability to detect differential protein expression, and the elucidated biological information. METHODS/STUDY POPULATION: Proteomic measurements of 103 clinically-annotated high-grade serous ovarian cancer (HGSOC) tumors previously genomically characterized by The Cancer Genome Atlas were conducted using two orthogonal mass spectrometry-based proteomic methods: iTRAQ and SWATH. The analytical differences between the two methods were compared with respect to relative protein abundances. To assess the ability to classify the tumors into subtypes based on proteomic signatures, an unbiased molecular taxonomy of HGSOC was established using protein abundance data. The 1,599 proteins quantified in both datasets were classified based on z-score-transformed protein abundances, and the emergent protein modules were characterized using weighted gene-correlation network analysis and Reactome pathway enrichment. RESULTS/ANTICIPATED RESULTS: Despite the greater than two-fold difference in the analytical depth of each proteomic method, common differentially expressed proteins in enriched pathways associated with the HGSOC Mesenchymal subtype were identified by both methods. The stability of tumor subtype classification was sensitive to the number of analyzed samples, and the statistically stable subgroups were identified by the data from both methods. Additionally, the homologous recombination deficiency-associated enriched DNA repair and chromosome organization pathways were conserved in both data sets. DISCUSSION/SIGNIFICANCE OF IMPACT: SWATH is a robust proteomic method that can be used to elucidate cancer biology. The lower number of proteins detected by SWATH compared to iTRAQ is mitigated by its streamlined workflow, increased sample throughput, and reduced sample requirement. SWATH therefore presents novel opportunities to enhance the efficiency of clinical proteomic studies.

OBJECTIVES/GOALS: Glioblastomas (GBMs) are heterogeneous, treatment-resistant tumors that are driven by populations of cancer stem cells (CSCs). In this study, we perform an epigenetic-focused functional genomics screen in GBM organoids and identify WDR5 as an essential epigenetic regulator in the SOX2-enriched, therapy resistant cancer stem cell niche. METHODS/STUDY POPULATION: Despite their importance for tumor growth, few molecular mechanisms critical for CSC population maintenance have been exploited for therapeutic development. We developed a spatially resolved loss-of-function screen in GBM patient-derived organoids to identify essential epigenetic regulators in the SOX2-enriched, therapy resistant niche. Our niche-specific screens identified WDR5, an H3K4 histone methyltransferase responsible for activating specific gene expression, as indispensable for GBM CSC growth and survival. RESULTS/ANTICIPATED RESULTS: In GBM CSC models, WDR5 inhibitors blocked WRAD complex assembly and reduced H3K4 trimethylation and expression of genes involved in CSC-relevant oncogenic pathways. H3K4me3 peaks lost with WDR5 inhibitor treatment occurred disproportionally on POU transcription factor motifs, required for stem cell maintenance and including the POU5F1(OCT4)::SOX2 motif. We incorporated a SOX2/OCT4 motif driven GFP reporter system into our CSC cell models and found that WDR5 inhibitor treatment resulted in dose-dependent silencing of stem cell reporter activity. Further, WDR5 inhibitor treatment altered the stem cell state, disrupting CSC in vitro growth and self-renewal as well as in vivo tumor growth. DISCUSSION/SIGNIFICANCE: Our results unveiled the role of WDR5 in maintaining the CSC state in GBM and provide a rationale for therapeutic development of WDR5 inhibitors for GBM and other advanced cancers. This conceptual and experimental framework can be applied to many cancers, and can unmask unique microenvironmental biology and rationally designed combination therapies.

OBJECTIVES/GOALS: In 2021, Duke University expanded the myRESEARCHsuite (MRS) of research support services with the launch of myRESEARCHpath (MRP), an interactive roadmap for navigating the project lifecycle. MRP integrates with the existing MRS services, which include a personalized research portal (myRESEARCHhome) and team of experts (myRESEARCHnavigators). METHODS/STUDY POPULATION: MRP was developed as a collaborative effort to centralize essential research-related information across Duke University into one location. MRP provides a web-based platform to integrate policies, processes, and resources from over 40 research support offices, organized into topic-based pages throughout the project lifecycle. Each topic-based page provides integrated guidance, categorized related resources, and contact information for personalized support from subject matter experts. Additional features of MRP include a curated search function, and filters that refine the topic-based pages and related resources to only those applicable to selected project inclusions and organizational unit. RESULTS/ANTICIPATED RESULTS: Since the launch of MRP in January 2021 through the third quarter of 2021, 5,947 unique users accessed MRP for a total of 17,452 sessions. The most commonly accessed topic-based pages during this time period were: Activity disclosures (Other Support and Current and Pending) – 3,231 pageviews Animal welfare – 1,882 pageviews Proposal review and submission – 1,306 pageviews NIH research grants (R series) – 686 pageviews Proposal planning – 669 pageviews The most frequently searched terms (including spelling variants) were Other Support, Biosketch, NIH, and no-cost extensions. DISCUSSION/SIGNIFICANCE: This data suggests users are accessing MRP for guidance on new or recently updated requirements. Maintaining clear, unified, and current site content should be prioritized to continue emphasizing MRP as a central location for research-related information. Duke also plans to explore further integration of MRP with the other MRS services.

OBJECTIVES/GOALS:

• Development of gold nanoparticles covalently linked to a photosensitizer for use to enhance radiation therapy. The particles will be thoroughly characterized structurally and mechanistically. The gold particles should enhance radiation activity by closer proximity to the photosensitizer and by increasing particle accumulation in the tumor.

METHODS/STUDY POPULATION:

• Gold nanoparticles were synthesized and coated with amine-terminated poly(ethylene) glycol, then covalently conjugated to chlorin e6, a known FDA-approved photosensitizer. The system was characterized using UV-Vis spectroscopy, transmission electron microscopy, and nanoparticle tracking analysis. The generation of reactive oxygen species was measured after X-irradiation. Enhanced cell killing was evaluated clonogenically in addition to assessment of in vivo efficacy and tumor pathology.

RESULTS/ANTICIPATED RESULTS:

• Conjugation of the particle to the photosensitizer was achieved, and the molecule was detected by UV-Vis spectroscopy. TEM and NTA showed no aggregation of the particles, and an increase in reactive oxygen species generation was observed. The conjugates increased cell killing during radiation treatment, whereas neither the particle alone nor the photosensitizer significantly affected clonogenic survival at the same concentrations. Breast tumors grown in immunocompetent mice showed increased necrotic tissue after a single 20 gy treatment in the presence of the conjugate.

DISCUSSION/SIGNIFICANCE OF IMPACT: Radiation therapy is widely used clinically, but dosage is limited largely to prevent injury to adjacent normal tissue. By increasing the local effect of radiation therapy, our gold conjugate has the potential to augment the effective radiation dose in the tumor, thereby reducing damage to healthy tissue and providing a more effective therapy.

ABSTRACT IMPACT: Lack of regulatory knowledge and education is a key barrier to the translation of medical devices and we describe the design and results for a university graduate-level course providing training on medical device regulatory submissions for approval that can help fill this unmet need and improve and accelerate translational success. OBJECTIVES/GOALS: Within the Indiana CTSI, the Medical Technology Advance Program (MTAP) in the Purdue University Weldon School of Biomedical Engineering (BME) offers three courses in regulatory science and regulatory affairs for medical devices. One course is focused on regulatory submissions for approval, and this report details the course design and evaluation. METHODS/STUDY POPULATION: For Fall 2020, the Regulatory Submissions for Approval course was enhanced to increase participation from regulatory professionals in US FDA and industry, with the core content, curriculum and course design led by BME faculty. The course was taught two days per week and included both in-person and remote (synchronous or asynchronous) attendance options. During the first class session each week a topic was covered in standard lecture format by BME faculty with industry regulatory experience. During the second class session, guests from both industry and FDA were invited to provide in-depth discussion on the topic, share perspectives and viewpoints, present real-world examples, experiences, and case studies, and answer student questions. An end of semester survey evaluated the effectiveness of the course design. RESULTS/ANTICIPATED RESULTS: Medical Device regulatory submissions and related activities were taught including product classification, presubmissions and meetings, 510(k), de novo, EUA, PMA, HDE, and advisory panels. FDA history, regulatory careers, regulatory science, and EU, China, and Japan regulations were also discussed. Overall, 29 speakers from FDA and industry participated live via video calls. A survey completed by 21/23 studentsrevealed overall satisfaction: all reported increased regulatory understanding and 20/21 learned ‘a lot’ or ‘an incredible amount’. The weekly lecture was the top factor contributing to learning, and guest speakers were the next most important factors. Nearly all students indicated FDA and industry speakers were ‘very’ or ‘extremely’ valuable/helpful. Additional results will be presented. DISCUSSION/SIGNIFICANCE OF FINDINGS: The three courses are designed to improve medical device translation by training students to better understand regulatory processes and pathways. Survey results and feedback indicated this course was successful. Continued participation from FDA and industry is critical to the learning. Additional case studies will also help enhance learning.

OBJECTIVES/SPECIFIC AIMS: Black women experience discrimination that targets their intersecting gender and ethnic identities, termed gendered racism (Essed, 1991). The gendered racism Black women experience has been linked to negative mental health outcomes (Thomas etal., 2008). The ‘strong Black woman’ is a cultural symbol of strength depicting the Black woman as unwavered by hardships, such as gendered racism (Shorter-Gooden & Washington, 1996). However, recent research suggests that belief in the strong Black woman cultural construct is associated with negative mental health outcomes (Watson & Hunter, 2015). The goals of the current study were to (1) replicate previous findings suggesting that experiences with gendered racism is positively correlated with psychological distress, (2) replicate previous findings suggesting that belief in the strong Black woman construct is positively correlated with psychological distress, and (3) explore how experiences with gendered racism and belief in the strong Black woman construct might interact to predict distress. METHODS/STUDY POPULATION: A national sample of 112 Black women completed an online survey via MTurk. Survey measures included the Gendered Racial Microaggressions Scale, Strong Black Woman Cultural Construct Scale, and Psychological Distress Scale. RESULTS/ANTICIPATED RESULTS: Pearson correlation revealed that experiences with gendered racism was positively correlated with psychological distress, r = 0.23, p = .02. Pearson correlation also revealed that belief in the strong Black woman cultural construct was positively correlated with psychological distress, r = 0.39, p < .001. Multiple linear regression revealed an interaction between experiences with gendered racism and belief in the strong Black woman construct (β = -0.18, p = .04) that predicted psychological distress, R2 = .20, F(3,108) = 8.63, p < .01. Namely, for those with high belief in the strong Black woman construct, experiences with gendered racism did not predict distress, β = -0.31, t = -0.29, p = .78. However, for those with low belief in the construct, experiences with gendered racism positively predicted distress, β = -2.57, t = 2.31, p = .02. DISCUSSION/SIGNIFICANCE OF IMPACT: The results underscore the harmful effects of gendered racism and gendered racial stereotypes on Black women’s mental health outcomes. Striving to appear as the strong Black woman is not likely to help Black women overcome daily hardships. In fact, belief in the strong Black woman construct is likely to add extra difficulties.

OBJECTIVES/GOALS: The primary research goal was to identify brain alterations reliably associated with obesity using coordinate-based meta-analysis. A secondary goal was to compare brain alterations in metabolically healthy (MHO) and unhealthy (MUO) obesity. METHODS/STUDY POPULATION: Source data were peer-reviewed studies reporting locations of gray-matter alterations in group-average, case-control contrasts (obese vs. non-obese) cohorts, performed in a whole-brain, voxel-wise manner. Both voxel-based morphometry and voxel-based physiology studies were included. Three coordinate-based meta-analyses were performed: Pooled (MUO + MHO), MHO, and MUO. RESULTS/ANTICIPATED RESULTS: Thirty-two studies reporting a total of 50 case-control contrasts (MHO, 23; MUO, 27) met inclusion criteria, representing 3,368 participants (obese, 1,781; non-obese, 1587). The pooled analysis yielded 8 cerebral foci (3 nuclear, 5 cortical) in regions implicated in reward-seeking, cognitive, and interoceptive behaviors. MHO yielded 7 cerebral foci (4 nuclear, 3 cortical), partially overlapping Pooled results, with similar behavioral loadings. The MUO pattern was distinct, with 3 cerebellar and 1 occipital foci. DISCUSSION/SIGNIFICANCE: Brain alterations occurred reliably in obesity. The dominant pattern (Pooled & MHO) involved cerebral reward-system circuits, evident even in metabolically healthy obesity. Cerebellar alterations occurred exclusively in metabolically unhealthy obesity, a pattern previously reported in metabolic syndrome.

OBJECTIVES/GOALS: CTSA Search Solutions (https://ctsa-search.rutgers.edu/search/) is a database that allows users to systematically conduct structured searches among the 60+ CTSA hub websites for information related to NCATS goals and CTSA hub activities. It was created with the objective of providing a novel process to evaluating and benchmarking CTSA hubs. METHODS/STUDY POPULATION: The CTSA Search Solutions database is an information tool that includes structured search terms relating to 3 main CTSA categories: NCATS goals, CTSA activities, and COVID 19 information. Subcategories from these topics were also identified and organized. Each CTSA hub website was systematically searched for content related to each of the identified terms and categories. The uniform resource locator (URL) for the primary webpage that provided content for each term was collected and stored in the CTSA Search Solutions database for user friendly access. URLs are validated monthly for changes or discrepancies. RESULTS/ANTICIPATED RESULTS: The final database includes access to 63 CTSA Hub websites with 89 structured search term options and over 800 links collected, organized, and published. Hub content can be searched by state, region, or even hub age to make detailed comparisons with the data identified. The CTSA Search Solutions tool allows researchers, administrators, evaluators, and community partners to find the needed links, to learn about specific CTSA hub program highlights as well as conduct research into program hub outputs and best practices across the nationwide CTSA continuum. DISCUSSION/SIGNIFICANCE: On the most practical level, CTSA Search Solutions has the potential to help hub evaluators identify the content of hubs in their first cycle compared to those in their 3rd Cycle. It can help core leads determine common best practices. It can help researchers understand the variety of COVID 19 outreach among the CTSA continuum.

OBJECTIVES/GOALS: The purpose of this study is to develop a clinically relevant mouse model of Radiation-Induced Heart Disease (RIHD) and characterize the resulting phenotype to find biomarkers and therapeutic targets as well as to understand the changes in cellular and molecular mechanisms of bioenergetics. METHODS/STUDY POPULATION: We used a two-beam method in the axillary region targeting the heart to irradiate male BALB/c mice at an isodose of 22, 16 and 8 Gray (Gy). We examined cardiac damage (i.e., vacuolization), inflammation, and DNA damage at 10 days post irradiation using histology and immunohistochemistry of heart tissue and cardiac function at day 35 by echocardiography. Additionally, cardiac tissue of mice irradiated at 22 Gy was collected at day 10 and day 35 post irradiation and sent for RNA sequencing. Data from RNA sequencing was analyzed using gProfiler, GSEA, and Cytoscape to enrich and visualize differentially expressed genes. RT-qPCR was performed to validate findings of significantly differentially expressed genes. RESULTS/ANTICIPATED RESULTS: Significantly increased phosphorylation of H2A.X indicated that irradiated mice were undergoing DNA double strand break repair indicating cardiac damage. Additionally, we found that regulators of mitochondrial function were decreased in the heart at day 10 for all doses. We found that mice that received 22 Gy developed cardiomyopathy at day 35 based on increased global longitudinal strain (GLS). Radiation decreased T cells, macrophages, and mast cells in the heart of irradiated mice by RT-qPCR at day 10 indicating damage to immune cells by radiation at all doses. Thus, we successfully created a clinically relevant model of RIHD in male BALB/c mice. DISCUSSION/SIGNIFICANCE: Patients undergoing radiation therapy for thoracic malignancies can develop cardiomyopathy (DCM) due to radiation damage. Previously published animal models utilized mouse strains resistant to developing DCM (female mice, C57BL/6 strain) and used high doses of radiation. Establishing a translational model is crucial for prevention of RIHD.

ABSTRACT IMPACT: We are developing the 3D perfusion system for use with patient-derived bacteria to further characterize the mechanism behind bacterial-induced inflammation and cancer. OBJECTIVES/GOALS: We previously reported the adherent invasive E. coli NC101 promote colorectal cancer (CRC) in mice. FimH, a mannose-specific adhesin on type 1 fimbriae, is involved in bacterial surface adhesion. Herein, we investigated the role of FimH in E. coli NC101-induced adherence and carcinogenesis in a novel 3D perfusion culture imaging plate. METHODS/STUDY POPULATION: E. coli NC101 gene fimH was deleted byï ŲRed Recombinase System. Biofilm formation was assessed by crystal violet and congo red staining. 5 dpf (wild-type strain) zebrafish embryos were infected in 6x107 cfu/ml wild type (WT) or fimH-deleted (ï ,,fimH) E. coli NC101 for 24hr and gut dissected for bacterial culture. A 2D/3D infection culture system for IEC-6 and HT-29 cells was infected for 4 hr and imaged and then DNA damage examined by comet assay, cell cycle andÎ3H2AX accumulation. Germ-free (GF) Il10-/- (colitis) mice were orally gavaged with 108 cfu WT orï ,,fimH E. coli NC101 for 16 weeks. E. coli colonization were quantified by plate culture and qPCR. Lipocalin2 was quantified by ELISA. PCNA and β-catenin were evaluated by immunohistochemistry (IHC). RESULTS/ANTICIPATED RESULTS: Biofilm formation was reduced by more than 40% (p<0.05) in E. coli NC101ï ,,fimH compared to WT strain. Zebrafish larvae showed a 41% decrease in intestinal colonization ofï ,,fimH compared to WT (p<0.05). E. coli NC101-induced DNA damage was reduced by 67% (p<0.0001) in HT-29 cells infected withï ,,fimH compared to WT strain. Using the 3D infection system, a 46% decrease in yH2AX (p<0.05) and 42% decrease in G2 cell cycle arrest (p<0.05) was observed inï ,,fimH infected IEC-6 cells compared to WT strain. Furthermore, ï ,,fimH infected Il10-/- mice showed decreased colonization (p<0.01), decreased intestinal inflammation (p<0.05), decreased stool lipocalin2 level (p<0.01), and reduction of PCNA positive cells in the intestine (p<0.05) compared to mice infected with WT strain. DISCUSSION/SIGNIFICANCE OF FINDINGS: Adhesin protein FimH is required by E. coli NC101 to colonize and promote colitis and carcinogenesis both in a 3D perfusion culture and in mice and may serve as potential therapeutic target.

Although mentoring is critical for career advancement, underrepresented minority (URM) faculty often lack access to mentoring opportunities. We sought to evaluate the impact of peer mentoring on career development success of URM early career faculty in the National Heart Lung and Blood Institute-sponsored, Programs to Increase Diversity Among Individuals Engaged in Health-Related Research-Functional and Translational Genomics of Blood Disorders (PRIDE-FTG). The outcome of peer mentoring was evaluated using the Mentoring Competency Assessment (MCA), a brief open-ended qualitative survey, and a semi-structured exit interview. Surveys were completed at baseline (Time 1), 6 months, and at the end of PRIDE-FTG participation (Time 2). The following results were obtained. Between Time 1 and Time 2, mentees’ self-assessment scores increased for the MCA (p < 0.01) with significant increases in effective communication (p < 0.001), aligning expectations (p < 0.05), assessing understanding (p < 0.01), and addressing diversity (p < 0.002). Mentees rated their peer mentors higher in the MCA with significant differences noted for promoting development (p < 0.027). These data suggest that PRIDE-FTG peer mentoring approaches successfully improved MCA competencies among URM junior faculty participants with faculty ranking peer mentors higher than themselves. Among URM faculty, peer mentoring initiatives should be investigated as a key strategy to support early career scholar development.

This special communication provides an approach for applying implementation science frameworks to a Clinical and Translational Science Institutes (CTSIs) community engagement (CE) program that measures the use of implementation strategies and outcomes that promote the uptake of CE in research. Using an iterative multi-disciplinary group process, we executed a four-phased approach to developing an evaluation plan: 1) creating an evaluation model adapted from Proctor’s conceptual model of implementation research; 2) mapping implementation strategies to CTSI CE program interventions that support change in research practice; 3) identifying and operationalizing measures for each strategy; and 4) conducting an evaluation. Phase 2 employed 73 implementation strategies across 9 domains generated by the Expert Recommendations for Implementing Change project. The nine domains were used to classify each CE program implementation strategy. In Phase 3, the group used the Reach, Effectiveness, Adoption, Implementation and Maintenance (RE-AIM) framework to define measures for each individual strategy. Phase 4 demonstrates the application of this framework and measures Year 1 outcomes for the strategy providing interactive assistance, which we implemented using a centralized consultation model. This approach can support the CTSA program in operationalizing CE program measurement to demonstrate which activities and strategies may lead to benefits derived by the program, institution, and community.

The COVID-19 pandemic changed the clinical research landscape in America. The most urgent challenge has been to rapidly review protocols submitted by investigators that were designed to learn more about or intervene in COVID-19. International Review Board (IRB) offices developed plans to rapidly review protocols related to the COVID-19 pandemic. An online survey was conducted with the IRB Directors at Clinical and Translational Science Awards (CTSA) institutions as well as two focus groups. Across the CTSA institutions, 66% reviewed COVID-19 protocols across all their IRB committees, 22% assigned protocols to just one committee, and 10% created a new committee for COVID-19 protocols. Fifty-two percent reported COVID-19 protocols were reviewed much faster, 41% somewhat faster, and 7% at the same speed as other protocols. Three percent reported that the COVID-19 protocols were reviewed with much better quality, 32% reported slightly better quality, and 65% reported the reviews were of the same quality as similar protocols before the COVID-19 pandemic. IRBs were able to respond to the emergent demand for reviewing COVID-19 protocols. Most of the increased review capacity was due to extra effort by IRB staff and members and not changes that will be easily implemented across all research going forward.