OBJECTIVES/GOALS: In 2020 the Georgia CTSA Clinical Research Center site at Emory University developed a highly trained, credentialed research coordinator pool with a goal to expand the pool to include clinical research coordinators from our partner institutions with the ability to work across institutional barriers in support of Georgia CTSA investigators. METHODS/STUDY POPULATION: Fall 2022, an Emory Investigator requested Georgia CTSA Biorepository samples with supporting clinical data for a NIH funded study. This provided a pilot opportunity to utilize clinical research nursing support offered by the UGA Clinical and Translational Research Unit (CTRU). De-identified samples were collected from our Biorepository while Emory’s coordinators and lab collaborated with UGA’s nursing support for data collection. Our obstacle for cross-institutional support was access to Emory Healthcare (EHC) medical records that would be needed by the UGA nurses, but partnerships created with the Georgia CTSA allowed us to overcome this, granting access to the electronic medical records (EMR) needed to complete the study. RESULTS/ANTICIPATED RESULTS: As expected, the process of credentialing and gaining access to the EHC EMR for the UGA team was the most time-consuming in the development of the pool. Discussions began in June 2021 to determine needs to allow the UGA research nurses to support the Emory coordinator pool. Requirements included acquiring an EHC network ID, completion of required Emory research training, letters of support from the Georgia CTSA outlining the collaboration between institutions, and a credentialing application. All steps were completed in May 2022 and the team began to identify studies that could benefit from this collaboration. Given that all credentialing and access needs were in place, the team was able to initiate the study and complete all study requirements, from sample identification to data collection and clean up, in five weeks. DISCUSSION/SIGNIFICANCE: Workforce shortages of experienced clinical research coordinators make it imperative to overcome barriers presented by institutional rules in order to efficiently utilize available resources to conduct high quality research. The CTSAs provide the perfect opportunity for partner institutions to develop processes to allow support across sites.

OBJECTIVES/GOALS: The goal of the Hatchery is a new approach to de-risk innovative life science ideas within an entrepreneurial setting. The Hatchery creates value by vetting the initial potential through experimental testing. METHODS/STUDY POPULATION: In this study we took a functioning wet laboratory space that was vacant due to principle investigator movement, and created a pipeline for its use in life science startup formation. A functioning laboratory can remain unoccupied for a notable period with transitions of research leadership. At the same time, a life science startup company who is testing core principles of their technology need wet lab space at an affordable cost. Our solution called the Hatchery provides startup companies a state-of-the-art wet laboratory space, next to a research hospital, for a very short duration of time and minimal fee. This novel approach allows preliminary validation of a technology for initial NIH SBIRs and STTRs funding pathways. RESULTS/ANTICIPATED RESULTS: Initial findings demonstrated the effectiveness of the Hatchery method. Our pilot study included five different life science startups company tenants. Each company was enabled to de-risk technologies and secure a phase 1 SBIR/ STTR funding or resulted in an exit via acquisition. DISCUSSION/SIGNIFICANCE: Entrepreneurship is a growing approach for testing and expanding new research areas. The Hatchery model makes use of existing space and infrastructure, can scale with an entrepreneurial community, and can serve as critical pilot data for a more permanent space commitment.

OBJECTIVES/GOALS: Evaluate effectiveness of participant recruitment service to improve study enrollment through customer feedback surveys and recruitment data obtained by research teams utilizing services. Use survey information to conduct gap analysis of additional support needed for participant recruitment. METHODS/STUDY POPULATION: Participant enrollment is often cited as one of the most challenging aspects of clinical research. In 2021, the Indiana CTSI used project management techniques to design and pilot a Team Science approach for providing participant recruitment support to clinical research teams. This service called the Indiana CTSI Recruitment Concierge Service (RCS) is comprised of recruitment, community engagement, social media, communications, and project management expertise. Additional experts are chosen to participate based on the study needs (regulatory, population insight, informatics, clinical services, etc.) RCS customers are comprised of study teams from a variety of areas and research experience. These customers are sent surveys to evaluate the support they received and provide suggestions for improvement. RESULTS/ANTICIPATED RESULTS: The RCS assisted 72 study teams in 2021 and 85 (as of November) in 2022. These studies were referred via word of mouth as no advertising of the service had been done to date. All customers were provided a study specific consultation with recommendations of services and resources that would assist their study. Some services recommended were: local study listing and volunteer registry (All IN for Health), digital marketing support, materials design expertise, community engagement and healthcare patient recruitment guidance. The overall feedback from RCS customers has been positive with most teams indicating the support improved their study recruitment and/or engagement plan. RCS will use information obtained to develop a strategy for prioritizing services due to the overwhelming number of requests received. DISCUSSION/SIGNIFICANCE: Using project management techniques and a Team Science approach, the Indiana CTSI was able to develop a comprehensive participant recruitment service that integrates clinical research operations, community engagement, and informatics expertise to design study specific recruitment plans and coordination of services.

OBJECTIVES/GOALS: Poor housing conditions and quality can be linked with residents’environmental exposures, which may contribute to a variety of adverse health outcomes. This systematic literature review will examine literature around housing, environmental exposure, and health; and policy implications to reduce the impact of housing on environmental exposures. METHODS/STUDY POPULATION: This systematic literature review will identify and evaluate published peer-reviewed articles as well as governmental and NGO policy briefs relating to connections between housing quality and condition, neighborhood characteristics, and environmental exposures (e.g., lead poisoning, secondhand smoke, PFAS chemicals) in the United States; and will particularly focus on health implications of such environmental exposures, racial/ethnic and socioeconomic disparities in exposure, and current and future policy recommendations to alleviate the association between housing and environmental risk. A computerized literature search of relevant electronic databases (e.g., PubMed, Sociological Abstracts, EPA database, Congressional database) for literature published after 2000 will be conducted. RESULTS/ANTICIPATED RESULTS: The findings from this literature review will be split up into categorizations around (1) the contribution of housing/neighborhoods on resident environmental exposure; (2) geospatial and demographic inequality (historic and current) around housing/neighborhood conditions contributing to disproportionate environmental exposures for low-income and minoritized residents; (3) health implications of environmental exposures; (4) prior policy addressing the connections between housing/neighborhoods and environmental risk; and (5) future policy recommendations to improve housing/neighborhood quality and minimize environmental exposures for residents. DISCUSSION/SIGNIFICANCE: This project will illuminate connections between housing conditions and environmental exposures, health implications of these exposures, and contribute to advancing understanding of potential policies to reduce adverse environmental health impacts of poor housing conditions for residents (particularly for low-income, minoritized groups).

OBJECTIVES/GOALS: To apply implementation science strategies to evaluate the impact of the 2016 Pennsylvania (PA) Medicaid payment policy change on hospital-level access to immediate postpartum long-acting reversible contraception (IPP LARC), an evidence-based strategy to increase contraceptive access; to identify differences by rurality and academic status. METHODS/STUDY POPULATION: We conducted a web-based, IRB-exempt survey of Labor and Delivery (L&D) leaders at all PA hospitals in Summer-Fall 2022, assessing hospital characteristics, contraceptive practices, and facilitators/barriers to IPP LARC implementation, using concepts from health services studies of small subsets of implementing hospitals; we translate these concepts into policy evaluation by sampling the complete population of Pennsylvania hospitals with active L&D units. L&D hospitals were characterized as sustainers if they implemented by 2019 and continued to provide IPP LARC, as implementers if they implemented IPP LARC in 2020-22, and non-implementers if they had not started the process. We use the Center for Rural Pennsylvania definition of rural: counties with RESULTS/ANTICIPATED RESULTS: We collected data from 48/74 (64.9%) hospitals with L&D units. Hospitals were heterogenous with 18/48 (37.5%) in rural counties and 15/48 (31.3%) identifying as academic. A minority of hospitals provide IPP LARC, with 17/48 (35.4%) offering implants and 16/48 (33.3%) offering intrauterine devices (IUD) immediately postpartum. Before the PA Medicaid payment policy change, few offered implants [4/48 (8.3%)] or IUDs [1/48 (2.1%)]. Non-rural hospitals implemented IPP LARC more often and on an earlier timeline than rural hospitals: [7/30 (23.3%) v 3/18 (16.7%)] sustainers, [5/30 (16.6%) v 2/18 (11.1%)] implementers. Common facilitators include clinical champions, meeting patient needs, and adequate knowledge. Planned analyses include implementation barriers, and impact of external implementation support. DISCUSSION/SIGNIFICANCE: Despite unmet need in rural populations for evidence-based contraception, rural hospitals were less likely to implement IPP LARC. Implementation support should be designed to meet the needs of rural hospitals. Implementation science methods can be translated to evaluate the impact of healthcare policy on access to care.

OBJECTIVES/GOALS: This investigation aimed to develop and validate a subject-specific finite element analysis (FEA) model with subject-specific mechanical loads during walking and to use this method to compare mechanical stresses between transfemoral and transtibial osseointegrated (OI) implants. METHODS/STUDY POPULATION: One patient with a unilateral transtibial OI prosthesis and one with a unilateral transfemoral OI prosthesis participated in motion analysis to collect kinematics and ground reaction forces during overground walking. Subject-specific musculoskeletal models were created, and static optimization was used to estimate muscle and joint reaction forces throughout walking. 3D FEA models of the tibia, femur, and implants were created using ScanIP and exported into ABAQUS CAE. Muscle forces were applied at corresponding origin/insertion locations, determined from the musculoskeletal models [7]. Fixed boundary conditions were applied at proximal joint centers, and bone stresses throughout gait were calculated. OpenSim and FEA derived estimates of joint reaction forces were compared for validation. RESULTS/ANTICIPATED RESULTS: A maximum stress of 65.53 MPa and 60.70 MPa was observed at the bone-implant interface for the transtibial and transfemoral patients (respectively) in the late stance phase of the walking task, corresponding to terminal stance and heel off. Averaged root mean squared errors of the walking task (in the anterior-posterior, inferior-superior, and medial-lateral directions, respectively) for the transtibial and transfemoral patients were (124, 152, 80) N, (71, 80, 30) N, and (190, 62, 30) N, respectively. DISCUSSION/SIGNIFICANCE: The purpose of the above study was to develop a methodology for determining subject-specific mechanical loads during walking using finite element analysis and compare mechanical stresses in patients with transfemoral and transtibial OI. Similar stresses between the two implant types were found.

OBJECTIVES/GOALS: Non-Typhoidal Salmonella causes over 95 million infections globally each year, and no effective vaccine exists to combat infections in humans. The goal of this study is to determine the immune protection provided by a novel extracellular vesicle (EV)-based vaccine generated using lab-strain Salmonella against wastewater-derived Salmonella. METHODS/STUDY POPULATION: We isolated Non-Typhoidal Salmonella (NTS) from raw influent wastewater samples collected from two wastewater reclamation facilities (WRF) in Gainesville, FL. Whole genome sequencing was performed on each isolate and compared to sequences of clinically-derived isolates in FL during our study period to identify a clinical and subclinical isolate for assessing EV based vaccine protection. Mouse serum and stool samples were collected from a cohort of EV-vaccinated mice. Surrogates of protection against Salmonella used anti-Salmonella IgA in the feces of these mice, and anti-Salmonella IgG in serum of the mice, by using ELISAs coated with whole cell lysate collected from the two wastewater-derived isolates. RESULTS/ANTICIPATED RESULTS: We have previously shown that an EV vaccine provides protection against Salmonella enterica Serovar Typhimurium, the serovar used in the generation of the EV vaccine. We anticipate that the EV vaccine generates additional protection against the community-acquired strains, which will be characterized by increases in fecal IgA and serum IgG against two community Salmonella isolates that is similar to responses against the serovar used to generate the EV vaccine (Typhimurium). DISCUSSION/SIGNIFICANCE: This study will improve the translation of our vaccine studies by demonstrating the efficacy of our novel EV vaccine against circulating Salmonella isolates.

OBJECTIVES/GOALS: Fetal glucose dynamics mediate many of the adverse outcomes seen in infants of diabetic mothers (IDM). The goals of this study are to identify: (1) rates of blood glucose change in normoglycemic and hypoglycemic IDM; (2) their relation to in-utero insulin exposure; and (3) their transcriptional impacts on placental and umbilical vasculature. METHODS/STUDY POPULATION: Using a longitudinal prospective study design, placental/umbilical cord tissue and maternal hemoglobin A1c (HbA1c) are being collected from mothers diagnosed with Type 1, Type 2, or gestational diabetes mellitus. Blood glucose levels are also collected from their infants at birth, and every 3-4 hours for up to 9 hours to determine the rate of change. Linear regression modeling will be used to determine associations between placental and umbilical endothelial RNA expression, umbilical cord insulin levels, and maternal HbA1c within each diabetic sub-type. Gene expression from endothelial specimens will be compared between diabetic sub-types and between normoglycemic and hypoglycemic infants via paired t-tests using Benjamini-Hochberg procedure for false discovery rate correction. RESULTS/ANTICIPATED RESULTS: We hypothesize the following; (1) glucose levels will have a steeper rate of change in hypoglycemic infants; (2) maternal HbA1c and in-utero insulin levels will correlate with the level of transcriptional change identified in placental and umbilical endothelial samples; (3) a negative association will exist between cord insulin levels and the rate of change in infant glucose levels; and (4) a positive association will exist between cord insulin level and transcriptional change on the placental and umbilical endothelium. DISCUSSION/SIGNIFICANCE: Identifying gene expression changes in diabetic placental/umbilical endothelium, and the role of insulin/glucose in these changes, is key to managing diabetic vasculopathy and its adverse outcomes. Understanding infant insulin response may also guide management of hypoglycemia and decrease the risk for neonatal intensive care unit admission.

OBJECTIVES/GOALS: Research on the safety of perinatal kratom use - an herb that acts on opioid receptors - is scarce. Our transdisciplinary clinical and translational science (CTS) team is conducting parallel qualitative analyses of subreddit posts related to kratom use during (1) pregnancy and (2) breastfeeding. METHODS/STUDY POPULATION: Pregnancy- and breastfeeding-related keywords are being used to extract posts and selected metadata from the following subreddit communities: r/kratom, r/quittingkratom, r/pregnant, and/or r/breastfeeding. After the removal of duplicate posts, posts written in a non-English language and those that state in the post text and/or title that they were published by minors ( RESULTS/ANTICIPATED RESULTS: Among the eligible posts, the number of unique usernames of the sources publishing the posts; the range of publication dates; and the mean, median, & range of the number of comments per post will be presented. Inter-rater concordance in thematic coding will be computed. A word cloud will be created with the most used nouns from the eligible posts. Verbatim quotes will be shown to illustrate themes depicted in the sample. The quantitative and qualitative analyses will be conducted separately for the posts related to kratom use during pregnancy and breastfeeding. DISCUSSION/SIGNIFICANCE: These findings could assist clinicians in identifying questions that obstetric patients may have regarding the perinatal use of this emerging substance of concern. Further research is needed to validate these findings using other social media data, such as Twitter.

OBJECTIVES/GOALS: Rodents are the most widely used experimental animals to study disease mechanisms due to their availability and cost-effectiveness. An international drive to investigate the pathophysiology of COVID-19 is inhibited by the resistance of rats and mice to SARS-CoV-2 infection. Our goal was to establish an appropriate small animal model. METHODS/STUDY POPULATION: To recreate the cytokine storm that is associated with COVID-19, we injected angiotensin converting enzyme 2 knockout (ACE2KO) mice (C57BI/6 strain) with lipopolysaccharide (LPS) intraperitoneally and measured the expression of multiple cytokines as a function of time and LPS dose. We then chose a minimum dose (500ug/kg) and time (3h) when multiple cytokines were elevated to measure lung injury scores using a point-counting technique on tissue sections stained with hematoxylin and eosin. The data are expressed as mean percentage of grid points lying within the peribronchial and superficial area in up to 20 fields. Percentage of peribronchial and superficial intrapulmonary hemorrhage, congestion, neutrophil infiltration and area of alveolar space were all assessed. RESULTS/ANTICIPATED RESULTS: Compared to the wildtype group (WT-G), the LPS-injected ACE2KO mice (LPS-G) exhibited a higher percentage of peribronchial intrapulmonary hemorrhage [(%): LPS-G, 10.56 ± 2.06 vs. WT-G, 5.59 ± 0.53; p DISCUSSION/SIGNIFICANCE: Establishing this novel mouse model of COVID-19 will facilitate studies investigating tissue-specific mechanisms of pathogenesis in this disease. This model can also be used to discover novel therapeutic targets and the design of clinical trials focusing on diagnostics, treatments and outcomes in COVID-19.

OBJECTIVES/GOALS: Selinexor is a novel XPO1 inhibitor that blocks nuclear export, thus impairing DNA repair and causing apoptosis. Our goal was to conduct preclinical and clinical studies to test our hypothesis that selinexor’s efficacy is boosted by priming with temozolomide and is associated with a tissue biomarker. METHODS/STUDY POPULATION: We leveraged a team science approach through the NCI Cancer Therapy Evaluation Program (CTEP) to design preclinical experiments, develop a novel RNAseq analysis pipeline, and use pre-existing clinical experience to open an early phase clinical trial for recurrent glioblastoma. Team members included a CTEP medical officer, cancer biologist, pharmacist, industry scientist, translational scientist, and early career clinician scientist mentored by an expert clinician scientist. Based on preclinical results, participants in the clinical trial experimental arm will receive sequential temozolomide 150mg/m2 on days 1-5 and a starting dose of selinexor 60mg on days 8 and 15 of a 28-day cycle. Participants in the control arm will receive monotherapy temozolomide. RESULTS/ANTICIPATED RESULTS: Sequential treatment of U87 cells and intracranial xenografts had superior DNA damage (É£H2A.X, cleaved PARP) and overall survival compared to combination or single-agent (HR 0.25 [95% CI, 0.07-0.84]; p=0.01, log-rank). We used the top-scoring gene pair method to identify an RNAseq signature associated with response to selinexor. We then designed a trial for first recurrent MGMT methylated glioblastoma. Primary objectives are safety and preliminary efficacy. Secondary objectives are overall response rate, efficacy, and validation of a molecular signature. Phase 1 dose finding (n=12) will be followed by a randomized phase 2 (n=72); using proportional hazards regression, RHR 0.5 with p DISCUSSION/SIGNIFICANCE: The NCI CTEP Project Team employs team science as a framework to successfully develop multidisciplinary collaborations, build investigator trial experience, and lead the way to future research opportunities. Our trial addresses a significant unmet need to offer novel therapies and molecular biomarkers in glioblastoma.

OBJECTIVES/GOALS: The goal of U-LINK is to bring together diverse scholars from multiple disciplines to address complex and challenging problems in healthcare, climate change, social equity, and community, through innovative team science, and aligned with the University of Miami’s strategic plans. METHODS/STUDY POPULATION: The U-LINK program has supported developmental and implementation projects through a competitive selection process. Developmental funding was intended for teams to develop and refine ideas and to become established as an effective team. Additional funding was provided to teams to advance their projects by conducting data collection and feasibility testing. In addition, U-LINK has supported fellowship for pre-doctoral and affiliated doctoral trainees. Team science training was provided to all teams through didactic lectures and hands-on training. Teams were tracked longitudinally by using surveys and bibliometrics to measure success and impact including scholarly output and follow on funding. Network analysis was performed to analyze research collaboration networks before and after ULINK funding. RESULTS/ANTICIPATED RESULTS: U-LINK has funded pilot programs and initial phases for 57 projects and 13 fellowships in the last three years. Over 400 individuals on teams from 16 schools/academic units collaborated on these projects on topics such as resilience, climate change, social equity and societal challenges, health, and impact of recent legislation on LGBTQ+ community. While data collection and analysis are ongoing, initial results show successful outcomes from U-LINK projects including publications and $29.5m in follow-on external funding. We anticipate network analysis to demonstrate increased and continued multi-disciplinary collaborations among U-LINK teams through co-authorship networks and increase in collaborative grants being submitted and/or funded. DISCUSSION/SIGNIFICANCE: The University of Miami’s U-LINK program has demonstrated success in forming interdisciplinary teams that have produced real-world solutions to complex problems by harnessing the inherent diversity and strength across UM’s programs.

OBJECTIVES/GOALS: This study aimed to investigate the mechanistic effects of the omega-6-derived eicosanoid 12-HETrE on platelets of cardiovascular patients at risk for a recurrent cardiovascular event triggered by thrombosis. 12-HETrE negatively regulates platelet reactivity through binding to the prostacyclin receptor in platelets. METHODS/STUDY POPULATION: Healthy donors were recruited from the Ann Arbor community and the University of Michigan Medicine Center. Cardiovascular patients with elevated cardiovascular risk were recruited from the Cardiac Catheterization Laboratory at Michigan Medicine Hospital. All subjects were recruited under study protocols approved by the University of Michigan IRB. Healthy donors were matched with the cardiovascular patients regarding age, sex, race, and BMI. Whole blood was collected via venipuncture into vacutainers containing sodium citrate. Platelets were isolated via serial centrifugation and treated ex vivo with vehicle control, 12-HETrE, or Iloprost. RESULTS/ANTICIPATED RESULTS: Based on our previous studies, we chose to treat platelets ex vivo with 25uM of 12-HETrE for 10 minutes. Using platelets of healthy donors, we have shown that treatment with 25uM of 12-HETrE for 10 minutes inhibited platelet aggregation and activation, and activated protein kinase A, suggesting activation of the prostacyclin receptor. We conducted a preliminary study to demonstrate that ex vivo treatment of 12-HETrE regulated signaling pathways in platelets such as cell-to-cell interaction, platelet activation and cytoskeleton rearrangements. In this study, we have demonstrated that treatment with 12-HETrE regulated receptors and intraplatelet proteins in platelets of cardiovascular patients. Furthermore, these proteins are involved in critical pathways in the platelet. DISCUSSION/SIGNIFICANCE: Dual antiplatelet therapy has significantly decreased mortality due to thrombotic events. However, cardiovascular events triggered by thrombosis persist as the leading cause of death in the US. This study may uncover key regulators to be targeted for the long-term goal of providing additional protection to reduce future incidence of thrombosis.

OBJECTIVES/GOALS: SLC6A1-Related Neurodevelopmental Disorder (SLC6A1-NDD) is a leading genetic cause of epilepsy and autism. Haploinsufficiency of SLC6A1 leads to reduced uptake of GABA from the synaptic cleft, and increased extracellular GABA in mice. It is a candidate for gene transfer therapy, but translational read outs are needed. EEG is a promising biomarker. METHODS/STUDY POPULATION: The SLC6A1-NDD program includes a specialty clinic and prospective cohort study run in parallel with pre-clinical gene therapy development. Characterization and pre-clinical testing of a homozygous knock-out, heterozygous knock-out, and two humanized knock-in models are on-going, including EEG analyses before and after treatment. Patients with a confirmed diagnosis of SLC6A1-NDD are seen annually in a specialty clinic and a subset participate in the cohort study which collects standardized questionnaires, EEGs, and MR Spectroscopy to measure glutamate and GABA. Gene Therapy Program investigators meet weekly to discuss progress on pre-clinical and clinical trial readiness on SLC6A1-NDD and to align efforts on translational read outs, including EEG, in both humans and the pre-clinical models. RESULTS/ANTICIPATED RESULTS: We have enrolled the full cohort of 20 participants in the prospective SLC6A1-NDD cohort study. Preliminary results have shown that all but 1 individual has a history of developmental delay, and 8 of the 24 individuals in our clinical cohort had at least one episode of developmental regression. Over 90% have epilepsy, and 17/20 in the cohort study have intermittent rhythmic delta activity on EEG. The full knock out mice have behavioral and learning deficits and abnormal electrical brain activity on telemetry, including bursts of spike trains, analogous to epileptiform activity seen in humans. Next steps include quantitative analysis of both mouse and human EEG to develop a translational brain-based biomarker. We plan to assess delta power and investigate genotype-phenotype correlations in mice and humans. DISCUSSION/SIGNIFICANCE: With targeted therapies in development for SLC6A1-NDD, translational biomarkers that demonstrate engagement with the brain are critical. With clinical heterogeneity in SLC6A1-NDD, biomarkers can objectively capture change. Collaborative translational projects may improve efficiency in rare disease research to facilitate early phase trials.

OBJECTIVES/GOALS: In 2020, Baylor College of Medicine held a datathon to introduce a data warehouse, identify its capabilities/limitations, foster collaborations, and engage trainees. The event was held again in 2022, and lessons learned (e.g., tools for data self-service or team communication) were applied. METHODS/STUDY POPULATION: Senior faculty reviewed proposals with an emphasis on feasibility, impact, and relevance to quality improvement or population health. Selected teams worked with Information Technology (IT) for 2 months and presented findings at a 1-day event. Surveys were administered to participants before and after the event to evaluate their background, team characteristics, collaborations, knowledge before and after the datathon, perceived value of the datathon, and plans for future work. Descriptive statistics of respondents’ self-reports were tabulated. RESULTS/ANTICIPATED RESULTS: In 2022, 19 of 36 projects were accepted (13/33 in 2020). At both events, most projects studied quality improvement or clinical outcomes. Of 82 participants in 2022, 54 completed surveys. In 2022, 72% had no datathon experience (48% in 2020). Median effort was 10 person-hours; median IT time was 20% (20 and 10%, in 2020). Seven respondents finished and 21 partially finished their projects (1 and 11, in 2020); 92% made new collaborations (91% in 2020). Respondents strongly agreed that: the experience was valuable (n=28), they would participate in future datathons (n=30), and they would use the warehouse for future work (n=25). Twenty-seven have planned abstracts; 25 have planned manuscripts. DISCUSSION/SIGNIFICANCE: The 2022 datathon had more participants with less experience, potentially due to improved promotion and training opportunities. Fewer person-hours and a higher percentage of IT time were required as compared to 2020, and more projects were completed, possibly due to increased IT efficiency.

OBJECTIVES/GOALS: Colorectal cancer (CRC) is a leading cause of cancer mortality, and many patients will develop metastatic disease at some point during their treatment course. Conventional therapies such as surgery, chemotherapy, and radiation are often of limited effectiveness in these advanced stages, which necessitates the development of novel therapies. METHODS/STUDY POPULATION: Our group has designed an oncolytic adenovirus backbone structure expressing the sodium iodide symporter (NIS) which can be used in conjunction with radioiodine to facilitate cancer imaging and therapy. Using multiple CRC cell lines, oncolytic adenoviruses with different fibers were tested in vitro to determine which of these modifications yielded the highest binding to the cancer cells. Additionally, multiple promoter structures are being tested to determine the impact on the replication and oncolytic effect of the virus. Furthermore, the potential of adenovirus-mediated NIS expression to facilitate PET/CT imaging and therapy with I-131 will be explored. RESULTS/ANTICIPATED RESULTS: The Ad5/3 chimeric fiber modification demonstrated the best binding in CRC cell lines. Additionally, tissue specific promoters are employed in oncolytic viruses to confer selective replication in cancer cells, while minimizing off target effects in nearby normal tissues. We have employed a Cox2 promoter, which has demonstrated an excellent oncolytic effect. In vitro NIS expression was shown in multiple CRC cell lines through immunostaining. Small animal PET/CT imaging demonstrated signal uptake in mice with subcutaneous CRC tumors after virus and radioiodine (I-124) administration. We anticipate that future studies employing radioactive iodine (I-131) in combination with our oncolytic virus will yield an augmented antitumor effect. DISCUSSION/SIGNIFICANCE: The NIS-expressing adenovirus has the ability to support radionuclide-based imaging and therapy for CRC. With additional pre-clinical testing, our adenovirus construct has the potential to bring NIS-based therapeutics to the bedside to positively impact CRC patient care outcomes.

OBJECTIVES/GOALS: Triple-negative breast cancer (TNBC) is a subtype of breast cancer that lacks effective targeted treatment options. TNBC's greater degree of immunogenicity than other breast tumors makes immunotherapy a viable strategy. Strategies to improve the immunotherapy response includes targeting the cGAS-STING innate immune pathway with STING agonists. METHODS/STUDY POPULATION: We have previously shown in vitro that eribulin, a microtubule destabilizer currently used in the treatment of TNBC, functions as an indirect STING agonist because it promotes the release of mitochondrial DNA into the cytoplasm. Separately, eribulin also significantly enhances type I interferon expression induced by STING agonists measured by qRT-PCR through a second TBK1-dependent mechanism downstream of STING activation through detecting higher amounts of phosphorylated IRF-3 by western blot protein analysis. Mechanisms of eribulin-mediated interferon expression occur in immune and TNBC cells and are shared with other microtubule destabilizers but not with the microtubule stabilizing agent paclitaxel. RESULTS/ANTICIPATED RESULTS: We determine that the enhancement of type I interferon expression by eribulin is pharmacologically synergistic with multiple STING agonists. The significant enhancement by eribulin led us to evaluate the antitumor efficacy of eribulin in combination the STING agonist ADU-S100 in a challenging spontaneous mammary tumor model MMTV-PyVT. We show that the combination treatment significantly decreased tumor growth which allowed for longer survival compared to other groups. This is particularly interesting because of our previous studies showing that eribulin alone, but not paclitaxel, promotes the activation of CD4+ T-cells in the spleen and draining lymph nodes of BALB/c mice with 4T1 tumors through flow cytometric analysis. DISCUSSION/SIGNIFICANCE: These data contribute to accumulating evidence that there are important mechanistic differences between the microtubule targeted chemotherapeutics currently used in the treatment of TNBC and suggest that eribulin can act as an immune adjuvant in addition to its anti-mitotic effect.

OBJECTIVES/GOALS: The goal of this study is to examine the incidence, etiology, and outcomes of hepatocellular carcinoma (HCC) in a 27-county region in SE Minnesota and W Wisconsin between 2010 and 2021. A comparison of the first to second half of the period will be made to look for possible trends. METHODS/STUDY POPULATION: The Rochester Epidemiology Project (REP) is a database of patient records across SE Minnesota and W Wisconsin. Starting in 2010, the REP opened to a 27-county catchment area, which includes over 1.3 million patients with a population coverage of approximately 64%. This study will use the expanded REP data to collect data on patients 20 years of age and older with a new diagnosis of HCC between Jan 1, 2010 and Dec 31, 2021—an estimated 1000 cases. Patients with a record of less than one year of residence in the catchment area will be excluded. Data on etiology, comorbidities, and outcomes of HCC will be extracted from medical records and analyzed for risk factors and changes over time. RESULTS/ANTICIPATED RESULTS: We anticipate that the overall incidence of HCC in the REP geographic area has increased over the period of 2010 to 2021. We anticipate that the prevalence of hepatitis C virus infection in patients with HCC has between 2010 and 2021, due to the widespread use and accessibility of hepatitis C-specific antiviral treatment over the past decade. We anticipate that the prevalence of NAFLD in patients with HCC has increased between 2010 and 2021. We do not anticipate significant changes in treatment modality or survival outcomes over this period. DISCUSSION/SIGNIFICANCE: This study will provide a comprehensive update on the state, etiology, and outcomes of HCC in the area surrounding Rochester, MN. We will be able to track changes in risk factors over time and work to enhance screening protocols to target the most vulnerable populations.

OBJECTIVES/GOALS: The characterization of the zebrafish as an animal model for Cockayne Syndrome may guide us towards role of Transcription-Coupled Nucleotide Excision Repair (TC-NER) defects in sensorineural hearing loss. METHODS/STUDY POPULATION: To examine our model, we have developed a zebrafish line with a 9+1 base-pair deletion in the ercc6 gene using TALENs. Mutation has since been confirmed by PCR and subsequent restriction digest with StuI. A series of assays evaluating hair cell morphology, structure and function, as well as ribbon synapse structure, will be used to analyze potential differences between the ercc6 mutant zebrafish line a their wild-type siblings. Additionally, electron microscopy will be used to assess differences in hair cell ultrastructure between the ercc6 mutant zebrafish line a their wild-type siblings. Finally, UVC exposure assays will be used to determine the role TC-NER plays in our novel zebrafish model, and evaluate its potential implications in sensorineural hearing loss. RESULTS/ANTICIPATED RESULTS: We anticipate that biallelic loss of function mutations in the zebrafish ercc6 gene will result in abnormalities in hair cell structure, mechanotransduction, or cell number. Additionally, we anticipate that hair cell ultrastructure and ribbon synapse structure will be impacted by loss of ercc6 expression. DISCUSSION/SIGNIFICANCE: Hearing loss mechanisms associated with defects in TC-NER are yet to be described. We believe our model will provide the tools for a faster and efficient way to carry out Cockayne Syndrome studies while laying the groundwork for the association between TC-NER and hearing loss.

OBJECTIVES/GOALS: The goal of this study was to develop a clinically applicable technique to increase the precision of in vivo dose monitoring during radiation therapy by mapping the dose deposition and resolving the temporal dose accumulation while the treatment is being delivered in real time. METHODS/STUDY POPULATION: Ironizing radiation acoustic imaging (iRAI) is a novel imaging concept with the potential to map the delivered radiation dose on anatomic structure in real time during external beam radiation therapy without interrupting the clinical workflow. The iRAI system consisted of a custom-designed two-dimensional (2D) matrix transducer array with integrated preamplifier array, driven by a clinic-ready ultrasound imaging platform. The feasibility of iRAI volumetric imaging in mapping dose delivery and real-time monitoring of temporal dose accumulation in a clinical treatment plan were investigated with a phantom, a rabbit model, and a cancer patient. RESULTS/ANTICIPATED RESULTS: The total dose deposition and temporal dose accumulation in 3D space of a clinical C-shape treatment plan in a targeted region were first imaged and optimized in a phantom. Then, semi-quantitative iRAI measurements were achieved in an in vivo rabbit model. Finally, for the first time, real-time visualization of radiation dose delivered deep in a patient with liver metastases was performed with a clinical linear accelerator. These studies demonstrate the potential of iRAI to monitor and quantify the radiation dose deposition during treatment. DISCUSSION/SIGNIFICANCE: Described here is the pioneering role of an iRAI system in mapping the 3D radiation dose deposition of a complex clinical radiotherapy treatment plan. iRAI offers a cost-effective and practical solution for real-time visualization of 3D radiation dose delivery, potentially leading to personalized radiotherapy with optimal efficacy and safety.