ABSTRACT IMPACT: Interdisciplinary networks represent critical components of translational science and learning system development. Our work impacts translational research by presenting an evidence-based approach to developing interdisciplinary networks in response to the COVID-19 pandemic; the approach presented may have broad applications within other academic institutions and medical centers. OBJECTIVES/GOALS: As a local response to the COVID-19 pandemic, we established the University of Alabama at Birmingham COVID-19 Collaborative Outcomes Research Enterprise (CORE) as an interdisciplinary learning health system (LHS) to achieve an integrated health services and outcomes research response amid the pandemic. METHODS/STUDY POPULATION: We adapted a learning system framework, based upon a scoping review of the literature and the Knowledge to Action Framework for implementation science. Leveraging this framework, we developed an institutional-level collaborative network of extant expertise and resources to rapidly develop an interdisciplinary response to COVID-19. The network was designed to quickly collect newly published or clinical information related to COVID-19, to evaluate potential usefulness of this information, and to disseminate the new knowledge throughout the interdisciplinary network; we strove to engage a wide variety of expertise and skills in the network. Thus, we subsequently used social network analysis to examine the emergence of informal work patterns and diversified network capabilities based on the LHS framework. RESULTS/ANTICIPATED RESULTS: We identified three principal characteristics of institutional LHS development including: 1.) identifying network components; 2.) building the institutional collaborative network; and 3.) diversifying network capabilities. Seven critical components of LHS were identified including: 1.) collaborative and executive leadership, 2.) research coordinating committee, 3.) oversight and ethics committee, 4.) thematic scientific working groups, 5.) programmatic working groups, 6.) informatics capabilities, and 7.) patient advisory groups. Evolving from the topical interests of the initial CORE participants, three scientific working groups (health disparities, neurocognition, and critical care) were developed to support the learning network. DISCUSSION/SIGNIFICANCE OF FINDINGS: Interdisciplinary collaborative networks are critical to the development of LHS. The COVID-19 CORE LHS framework served as a foundational resource that may support further institutional-level efforts to develop responsive learning networks. The LHS approach presented may have broad applications within other academic institutions and centers.

Objectives/Goals: The never in mitosis kinase (NEK) family regulates vital processes, namely cell cycle progression, but their potential as therapeutic targets in TNBC has not been fully explored. Our studies aim to develop a toolkit to investigate the functional roles of NEKs in pathologies including carcinogenesis. Methods/Study Population: To assess differential NEK expression in normal and tumor tissues and correlation of gene expression with patient survival, we used Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan–Meier Plotter (KMPlot) pan-cancer analysis, respectively. Basal NEK protein levels were determined by immunoblot across a panel of cell lines, including breast cancer, osteosarcoma, hepatocellular carcinoma, and non-cancerous cells, to identify appropriate systems for evaluation of NEK function. Doxycycline-inducible cell lines were generated by transduction with lentiviral stocks of NEK shRNA and overexpression constructs and antibiotic selection. Expression was analyzed by qPCR and immunoblot. Results/Anticipated Results: Expression of NEK2, 4, 5, 6, 8, and 11 was higher in breast tumors compared to normal tissue by GEPIA analysis. Further examination using KMPlot showed a correlation between elevated NEK6 expression and decreased overall survival in patients with aggressive cancers. As an initial proof-of-concept study, we analyzed NEK6 protein expression in breast cancer cells. Levels of NEK6 were elevated in TNBC cells (MDA-MB-231) compared to hormone receptor positive (HR+) breast cancer cells (MCF7). Using complementary approaches to investigate the functional role of NEK6 in breast cancer, we depleted NEK6 expression using shRNAs in TNBC cells and expressed NEK6 in HR+ cells Discussion/Significance of Impact: Because kinase dysregulation promotes oncogenesis and metastasis, targeting kinases is a key strategy in therapeutic development. A NEK-specific molecular toolkit allows researchers to elucidate NEK functions and contributions to carcinogenesis, promoting advancement of novel therapies.

Objectives/Goals: The primary objective of this study is to test whether NAD+ modulation lowers mitochondrial oxidative stress to restore endothelial nitric-oxide signaling and limit microvascular remodeling in diabetes in murine and 3D tissue-engineered models. The goal is to define NAD-responsive pathways and candidate blood biomarkers of microvascular injury. Methods/Study Population: Use db/db diabetic mice to measure effects of NAD+ modulation on capillary density, perfusion, endothelial nitric oxide (NO) signaling, and mitochondrial reactive oxygen species (ROS) by histology, immunohistochemistry (IHC), qPCR/immunoblot, and functional assays. Build a 3D microfluidic model of diabetic retinal capillary disease from human endothelial/pericyte co-cultures with a retina-on-a-chip model; induce diabetic stress (hyperglycemia + inflammatory cytokines) and apply an NAD+ modulation to map dose–responses for barrier integrity, ROS, NO, and morphology. Analyze publicly available transcriptomic datasets from people with and without diabetic microvascular disease to identify oxidative-stress signatures and candidate circulating biomarkers. Results/Anticipated Results: Expect increasing NAD+ availability to reduce mitochondrial ROS and inflammatory signaling, increase SIRT3/SOD2 activity, improve NO bioavailability, and prevent capillary rarefaction and leak in db/db mice. In the microfluidic model, anticipate a rescue of barrier function, lumen stability, and branching with NAD+, defining an effective dose window. Cross-platform analyses should converge on NAD+-responsive pathways (redox, mitochondrial metabolism, endothelial junctions) and nominate blood biomarkers that correlate with microvascular status. Together, results will provide actionable preclinical evidence for NAD+ modulation as a therapeutic strategy. Discussion/Significance of Impact: By targeting a shared driver, mitochondrial oxidative stress, this work could deliver a mechanism-based therapy and practical biomarkers for diabetic microvascular disease, accelerating translation from bench models to patient-focused trials and improving cardiovascular 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.

ABSTRACT IMPACT: In light of the high co-occurrence between intimate partner violence (IPV) and physical child abuse, studying these forms of aggression simultaneously, bidirectionally, and longitudinally is vital to address this public health need. OBJECTIVES/GOALS: This study examined reciprocal associations between parent-child aggression (PCA) risk, IPV victimization, and perceived child behavior problems, to evaluate whether negative processes can transmit across family subsystems (i.e., spillover hypothesis) and/or across individuals (i.e., crossover hypothesis) over time. METHODS/STUDY POPULATION: Participants were first-time mothers and their male partners enrolled in a prospective longitudinal study, which tracked parenting and PCA risk over the transition to parenthood and into early childhood. The current project examined data from the third and fourth waves, when participants’ child was 18 months old and approximately 4 years old. At both timepoints, parents reported on their PCA risk (i.e., child abuse potential, harsh parenting), physical and psychological IPV victimization and perpetration, and perceived child behavior problems. Mothers and fathers each completed protocol on laptops in separate private rooms. Hypotheses were tested with autoregressive cross-lagged path models, which were estimated for mothers and fathers separately as well as dyadically. RESULTS/ANTICIPATED RESULTS: Findings partially supported the hypotheses, with evidence of spillover occurring bidirectionally for mothers and unidirectionally for both mothers and fathers. Mothers’ PCA risk predicted their subsequent IPV victimization and their reported child behavior problems (i.e., spillover effects) as well as fathers’ reported IPV victimization (i.e., crossover effect). Maternal reports of child behavior problems also predicted mothers’ reported IPV victimization and fathers’ reported child behavior problems, indicating possible child-driven effects. Overall, results demonstrate the mutual influence of individuals and subsystems within the family. Additionally, mothers rather than fathers appear more vulnerable to harmful spillover effects. DISCUSSION/SIGNIFICANCE OF FINDINGS: The need for family prevention and intervention services is clear, given the complex, transactional nature of family violence. Particularly for mothers, higher PCA risk may herald an increased risk for subsequent IPV victimization and vice versa. Clinical implications for parent-focused intervention programs are discussed.

OBJECTIVES/GOALS: Metastasis to regional areas decreases invasive breast cancer (IBC) survival rate by 13%. Despite the clinical importance of lymph node involvement, the role of extracellular matrix (ECM) remodeling in metastases is unknown. We hypothesize that the spatial dysregulation of the collagen proteome facilitates pro-tumorigenic immune infiltration. METHODS/STUDY POPULATION: Lymph node metastases were compared to patient-matched primary tumor and normal lymph nodes using tissue microarrays (TMA) from 31 generational South Carolina women with IBC (black women, BW n=10, white women, WW n=21) and lumpectomies from 5 triple-negative breast cancer (TNBC) patients (BW n=3; WW n=2) by ECM-targeted mass spectrometry imaging. RESULTS/ANTICIPATED RESULTS: Between metastatic and normal lymph nodes, 10% of peptides, primarily from fibrillar collagens, were significantly different by area under the receiver operating curve (AUROC>70%; p-value< 0.01) within the TMAs. In a subsequent preliminary study of the TNBC metastatic niche, a segmentation analysis of 152 putatively identified peptides and 117,909 pixels revealed 10 uniquely localized proteomic groups. 12 peptides were found to have significantly decreased relative peak intensities in lymph node metastases compared to the primary tumor and normal lymph nodes by a one-way ANOVA test (p< 0.05). 7 peptides could discriminate between metastatic and normal lymph nodes, while 22 peptides could discriminate between metastatic lymph nodes and the primary tumor (AUROC>0.70; p-value < 0.05). DISCUSSION/SIGNIFICANCE: Our preliminary interrogation highlights emerging differences between lymph node metastases, the primary tumor, and normal lymph nodes. Future work is needed to connect these discrete ECM proteomes to immune infiltration alterations, which could contribute to disparate patient outcomes.

Objectives/Goals: To develop and deploy an academic learning health system (aLHS) Bridge Program to capitalize on our unique organizational strengths in Implementation Science (IS) and to overcome the gap between science and practice that threatens the success of an aLHS. The aLHS Bridges includes an IS Shared Resource, intended to advance IS to practice. Methods/Study Population: The new IS Shared Resource is built on our expertise in dissemination and implementation science, pragmatic, and adaptive trials and the CTSI’s prior success in integrating academic and clinical missions. We also leveraged our existing experts to co-lead the aLHS Bridge including Kristie Foley, PhD, inaugural Chair of the Department of Implementation Science, and Gary Rosenthal, MD, Chair of the Department of Internal Medicine. Specifically, the new IS Shared Resource builds on the capacity of the Department of IS, comprised of 36 faculty members (19 primary and 17 secondary/adjunct appointments) with expertise in qualitative and mixed-methods research, stakeholder engagement, participatory research, digital health, and organizational theory. Results/Anticipated Results: The IS Shared Resource is primed to aid faculty with dissemination and implementation needs, including shortening the time of intervention adoption and using implementation science to inform sustainable and effective implementation practice. The IS Shared Resource is equipped to provide consultation services to faculty members to understand their specific request and match IS faculty members who are expertly trained in specific strategies or contexts. Discussion/Significance of Impact: Leveraging current resources and our first-of-its-kind Department of Implementation Science, our CTSI was able to stand up the IS Shared Resource to support the goals of the CTSA and our greater institution mission. Using a multidisciplinary approach was essential to the success of the IS Shared Resource.

OBJECTIVES/GOALS: Non-tuberculous mycobacteria (NTM) is an emerging infection in the United States. Therapeutic development is impaired in NTM due to lack of biomarkers associated with disease activity and treatment response. To address this, we established a cross-sectional patient registry and biorepository from NTM patients. METHODS/STUDY POPULATION: Beginning August 2019 patients were recruited in a cross-sectional format from the bronchiectasis and NTM clinics at our institution. All patients provided at least one sputum sample in the six months prior to inclusion. Clinical and epidemiologically relevant data was obtained, patient reported outcome measures including the SGRQ were provided, and blood specimens were processed and preserved. Patients were grouped based on clinical phenotype and descriptive statistics were reported as means and standard deviations. Serum inflammatory profiles will be analyzed using standard Luminex assays. RESULTS/ANTICIPATED RESULTS: 72 patients with prior NTM isolation from sputum have been recruited including 28 patients that do not meet ATS guidelines for NTM treatment (colonized), 29 patients are currently receiving treatment, and 15 patients that have a history of completing therapy. Among all NTM patients, the mean age was 59.5 ±17.6 years and 80.8% were female. The mean FEV1 percent predicted among these patients was 68.0 ±21.5% and mean BMI was 22.35 ±4.1. The most common mycobacterial species isolated was Mycobacterium Avium Complex (47%). The mean SGRQ total score was 40.2 ±20.3 among all NTM patients. We plan to perform standard Luminex assays to identify inflammatory profiles associated with disease phenotype. DISCUSSION/SIGNIFICANCE OF IMPACT: We developed a cross sectional cohort of NTM patients including a registry of clinically relevant data for phenotyping and an accompanying biospecimen repository. Our long-term goal is to identify biomarkers indicative of disease activity and treatment response using these components.

OBJECTIVES/SPECIFIC AIMS: Evaluate how access and affordability has changed before and after the implementation HIEs in three subpopulations. The subpopulations are individuals who are currently insured through the HIE but were previously: 1. Insured through Employment-based insurance (PEBI subpopulation) 2. Insured through Private Insurance (PPI subpopulation) and 3. Uninsured (PU subpopulation). The three access and affordability measures are: Outcome measure 1. Did not fill a prescription in the past year due to cost Outcome measure 2. Could not get needed medical exam in the past year due to cost and Outcome measure 3. Had problems paying medical bills in the past year. METHODS/STUDY POPULATION: We analyzed the de-identified public use data from the 2012 and 2015 Ohio Medicaid Assessment Survey (OMAS). Sponsored by the Ohio Department of Medicaid, Ohio Department of Health, and the Ohio State University, the OMAS is a representative cross-sectional survey of non-institutionalized Ohio residents, regardless of their Medicaid status. In order to “longitudinalize” the 2012 and 2015 cross-sectional data of the OMAS, we employed a propensity score-based approach. We started with the 2015 OMAS, and carefully characterized each of the PEBI, PPI, and PU subpopulations along 17 demographic, health utilization, health behavior, and health status covariates using a propensity score model. Then, we identified controls for the three subpopulations within the 2012 OMAS data using the propensity scores. Finally, we estimated the odds ratios for the three outcome measures between 2012 and 2015. RESULTS/ANTICIPATED RESULTS: In 2015 there were approximately 201,381 adults (unweighted count = 996) who were insured through the HIE in Ohio. Of those individuals, 17.7% fell into the PEBI subpopulation, 17.6% fell into the PPI subpopulation, and 53.3% fell into the PU subpopulation; the balance of the respondents (11.4%) reported previously having Medicaid, or “Other” insurance. There are several key differences in the covariates at baseline between the three subpopulations. In general, the PU subpopulation tended to younger, more minority, more socioeconomically disadvantaged, and more likely to not have a primary care provider compared to both the PEBI and PPI subpopulations. In the 2012 data, we were able to identify 170 controls for the PEBI subpopulation, 167 controls for the PPI subpopulation, and 516 controls for the PU subpopulation. Compared to 2012, in 2016 (after the implementation of the HIEs):. Outcome measure 1: The PEBI subpopulation was more likely to report not filling a prescription in the past year due to cost (there were no significant changes in the PPI or PU subpopulations). Outcome measure 2: The PEBI subpopulation was more likely to report not getting a needed medical exam or medical supplies in the past year due to cost. The PPI subpopulation was less likely to report not getting a needed medical exam or medical supplies in the past year due to cost. There were no significant changes for the PU subpopulation for this outcome measure. Outcome measure 3: There were no changes in the “had problems paying medical bill in the past year” outcome across the three subpopulations. DISCUSSION/SIGNIFICANCE OF IMPACT: This is among the most detailed studies of health insurance exchanges known to the investigators. Analyzing outcomes at the subpopulation level illustrates that there have been unbalanced gains in access and affordability as a result of the HIEs. In general, those who were previously insured through employer-based insurance saw their access and affordability decrease; those previously insured through private insurance saw modest increases to access and affordability; and perhaps most surprising, those that were previously uninsured saw no changes to their access and affordability. Future studies will incorporate 2017 OMAS data (when it becomes available) to see if these trends persist over time. During this time of rapid health systems and health policy change, our study adds an important contribution to the discussion surrounding how to best deliver highly effective and efficient health care.

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: 1. To determine older adults’ opinions on content that is valuable for inclusion in a falls-prevention self-management plan. 2. To determine older adults’ recommendations of mode(s) to promote adherence to falls prevention recommendations. METHODS/STUDY POPULATION: On-on-one semi structured interviews with older adults are ongoing to determine their opinions on content for inclusion in a falls-prevention self-management plan and recommendations for mode of delivery. As in our prior investigations, we used the theoretical constructs of the health belief model to develop our questions. Interviews will be recorded and transcribed. Data will be entered into MAXQDA12 and coded. Concurrent data collection and analysis will continue until theoretical saturation of themes are achieved. Through this iterative process, we will identify content and mode of delivery for a falls- prevention self-management plan for implementation with older adults. RESULTS/ANTICIPATED RESULTS: We anticipate we will have conducted enough interviews to achieve data saturation by February, 2020. We expect the results of this qualitative investigation to guide the development of a falls-prevention self-management plan that includes targeted implementation and adherence strategies deemed acceptable and feasible for use among older adults following community-based falls-risk screenings. DISCUSSION/SIGNIFICANCE OF IMPACT: Falls are the leading cause of morbidity and mortality among older adults. Adherence to falls prevention among older adults is poor, even among those that voluntarily seek out recommendations. The results of this study will assist with development and pilot testing of a falls-prevention self-management plan to assist older adults to adhere to recommendations.

ABSTRACT IMPACT: The findings of this study could lend us insights into behavioral intervention that could potentially prevent or slow the onset of Alzheimer’s disease. OBJECTIVES/GOALS: This study examines the association between cognitive and physical resilience and Alzheimer’s disease in a Tibetan Buddhist monastic community in southern India. METHODS/STUDY POPULATION: The study will employ mixed methods of semi- and unstructured interviews and surveys. The interviews will be conducted among 60 monks of age 50+ in six Tibetan monastic colleges in southern India. The interviews will comprise general questions related to monks’ monastic educations and practices, as well as clinical cognitive interviews. Interviewees will be randomly sampled from a census of monks at the six monasteries. Owing to COVID-19 crisis, we will begin data collection, starting with interviews via zoom in mid-December 2020. The survey, which includes demographic information, cognitive assessments, meditative practices, health, memory and physical activity, will be conducted among 400 monks. The survey will be performed onsite and is tentatively scheduled in the summer of 2021. RESULTS/ANTICIPATED RESULTS: The study will help to identify factors associated with physical and cognitive resilience and develop measures to quantify and describe meditative and cognitive practices. These data will be used to both adapt validated measures developed in Western populations for use with this unique population and to develop new items on physical and cognitive resilience to include in the planned survey. Furthermore, the study will provide information about the prevalence of Mild Cognitive Impairment and Alzheimer’s disease and related dementias in this population and development of the survey to capture culturally appropriate measures, including on meditation. The findings could eventually lend us insights into behavioral intervention that could potentially prevent or slow the onset of Alzheimer’s in wider population. DISCUSSION/SIGNIFICANCE OF FINDINGS: Cognitive and physical resilience are understood to confer significant benefits to health outcomes and healthy aging. However, the factors related to resilience, particularly in older adults, are poorly understood. This study will estimate the link between frequency and intensity of meditative practices and physical and cognitive resilience.

Objectives/Goals: Our objective is to develop and validate a human iPSC-derived 3D osteosarcoma (3D-iOSA) model that replicates chromosomal instability (CIN), metastasis, and chemoresistance. The model will serve as a platform to evaluate chemotherapy and immune cell therapeutics, offering improved predictive fidelity compared to 2D and spheroid systems. Methods/Study Population: Human iPSCs were engineered by CRISPR/Cas9-mediated knock out of TP53 and RB1, differentiated to mesenchymal stem cells with overexpression of cMYC ± hRAS. Cells were embedded in hyaluronan and osteogenically differentiated to generate 3D organoids (3D-iOSA), with osteoblastic commitment confirmed by RUNX2 and BGLAP immunohistochemistry (IHC). Tumorigenic and metastatic potential was assessed via subcutaneous implantation into immunodeficient mice, followed by histopathology, IHC, RNA sequencing, and 5X whole genome sequencing (WGS). MAP (methotrexate, doxorubicin, and cisplatin) chemotherapy response and chimeric antigen receptor (CAR)-immune cell cytotoxicity were compared across 2D and 3D systems to evaluate chemoresistance, immune evasion, and translational fidelity of the 3D-iOSA model. Results/Anticipated Results: 3D-iOSA exhibited osteogenic lineage commitment with RUNX2 and BGLAP expression and disorganized, anaplastic morphology correlated with mutational burden. Tumors displayed greater histologic complexity than 2D-derived tumors, with pulmonary metastases in 50% of primary and 70% of secondary recipients. WGS revealed recurrent copy number alterations and subclonal variation, indicating ongoing CIN. MAP half-maximal inhibitory concentration values were 106-260× higher in 3D-iOSA organoids than 2D cultures, increasing with genetic complexity. B7H3 CAR-iPSC NK-cell and B7H3/HER2 dual-CAR γδ T-cell screening showed system-dependent efficacy, with reduced cytotoxicity in 3D-iOSA organoids versus 2D and spheroid cultures, highlighting 3D-iOSA superior predictive power for immunotherapy evaluation. Discussion/Significance of Impact: 3D-iOSA is the first human OSA model to replicate CIN-driven evolution, chemoresistance, and metastasis. It distinguishes differential immune therapy responses with greater predictive power than 2D or spheroid models, providing a scalable, physiologically relevant platform to accelerate targeted therapy development.

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/SPECIFIC AIMS: We hypothesize that VL testing varies by geographic sub-region, country, age, gender, mode of transmission, year of diagnosis, and country of origin; and also that a higher prevalence of VL testing may be associated with higher prevalence of population-level VL suppression. Our primary aim is to determine country- and regional-level factors that are associated with viral load testing amongst HIV patients. Our secondary aim is to explore the association between prevalence of viral load testing and viral load suppression at the population level. METHODS/STUDY POPULATION: This is a retrospective analysis of de-identified individual-level data reported to the European Surveillance System (TESSy). The TESSy is a database of communicable diseases (including HIV) for the ECDC and WHO European Regional Office. It captures data from 31 European Union/European Economic Area (EU/EEA) countries and 23 non-EU/EEA countries. Stored data is from year 2000. TESSy is used for data analysis and production of outputs for public health action. The patient cohort include adults older 18 years, whose last clinic attendance was reported in 2014 or later, or whose viral load test was reported in the year of the visit or the year before the year of their last reported clinic attendance. Patient demographic data include age, sex, mode of transmission, country of origin (migrants), country of diagnosis, geographic region, last clinic attendance, viral load and therapy status. Geographic region will be categorized into East, West and Centre as per WHO guidelines. Countries will be categorized and analyzed according to their European Union (EU)-, European Economic Area (EEA)- and income (GDP)-status, using current World Bank and International Monetary Fund (IMF) guidelines. All statistical analysis will be performed in R-Studio and R i386 3.0.2. Missing data will be characterized in terms of quantity (how much is missing) and pattern (random versus non-random) and impact on covariates to be tested. Multiple data imputations would be used in cases where missing data is found to be at random. Data from external sources like UNAIDS, World Bank and IMF will also be used for comparison and validation of TESSy data for imputation of missing data. Continuous variables will be analyzed through appropriate parametric and non-parametric tests while categorical variables will be analyzed through methods of proportion. Multivariate logistic regression methods will be used to explore the associations between VL testing and VL suppression separately with age, sex, year of diagnosis, country of origin (migrants), mode of transmission, in the total population, then at country- and regional-level. The same associations will be explored using a country’s EU and EEA status (EU versus EEA versus non-EU/EEA), and income status (high versus upper middle versus lower middle versus low). DISCUSSION/SIGNIFICANCE OF IMPACT: Even though this is a retrospective analysis of a database with likely significant missing data that may affect analysis of data and interpretation of results, our study will impact all levels of HIV policy across Europe. The strengths of this study likely outweigh the limitation imposed by missing data and include potential regional-, country- and demographic-specific public health, epidemiologic and ART program policy initiatives. Also our analysis of pattern of missing data may inform a more efficient and meaningful data collection and input into TESSy database.

OBJECTIVES/GOALS: Active surveillance (AS) is a recognized strategy to manage low-risk prostate cancer (PCa) in the absence of cancer progression. Little prospective data exists on the decisional factors associated with selecting and adhering to AS in the absence of cancer progression. We developed a survey instrument to predict AS uptake and adherence. METHODS/STUDY POPULATION: We utilized a three-step process to develop and refine a survey instrument designed to predict AS uptake and adherence among men with low-risk PCa: 1) We identified relevant conceptual domains based on prior research and a literature review. 2) We conducted 21 semi-structured concept elicitation interviews to identify patient-perceived barriers and facilitators to AS uptake and adherence among men with a low-risk PCa who had been on AS for ≥1 year. The identified concepts became the basis of our draft survey instrument. 3) We conducted two rounds of cognitive interviews with men with low-risk PCa (n = 12; n = 6) to refine and initially validate the instrument. RESULTS/ANTICIPATED RESULTS: Relevant concepts identified from the initial interviews included the importance of patient: knowledge of their PCa risk, value in delaying treatment, trust in urologist and the AS surveillance protocol, and perceived social support. Initially, the survey was drafted as a single instrument to be administered after a patient had selected AS comprising sections on patient health, AS selection, and AS adherence. Based on the first round of cognitive interviews, we revised the single instrument into two surveys to track shifts in patient preference and experience. The first, administered at diagnosis, focuses on selection, and the second, a 6-month follow up, focuses on adherence. Following revisions, participants indicated the revised 2-part instrument was clear and not burdensome to complete. DISCUSSION/SIGNIFICANCE OF IMPACT: The instrument’s content validity was evaluated through cognitive interviews, which supported that the survey items’ intended and understood meanings were isomorphic. In the next phase, we plan to conduct a large-scale prospective cohort study to evaluate the predictive validity, after which it will be available for public research use.

OBJECTIVES/GOALS: Uterine fibroids are benign tumors of the uterus with a high disease prevalence and burden, yet there are few multi-ancestry genetic studies. This is the largest and most diverse fibroid GWAS to-date. Our goal is to identify novel genetic variants and gene expression pathways associated with fibroids and characterize their biological relevance. METHODS/STUDY POPULATION: We performed a cross-ancestry meta-analysis of GWAS summary statistics from eight datasets. The total sample size was 74,294 cases and 465,810 controls with participants of European (80% of sample), African (4%), East Asian, and Central South Asian (16%) ancestry. We mapped variants to genes with OpenTarget Genetics and used Functional Mapping and Annotation to conduct tissue expression gene-set enrichment and identify lead variants. We used S-PrediXcan to estimate genetically predicted gene expression (GPGE) associated with fibroid risk. This was with models that predicted gene expression across 49 different tissue types. Ingenuity Pathway Analysis compiled significant GPGE genes and their weights with a scientific literature database to identify overlapping pathways. RESULTS/ANTICIPATED RESULTS: We identified 370 independent significant variants. Among these, we identified variants mapped to three novel genes (PAX2, VIP, FOXO3) and eight genes not previously validated (TEKT1, SLC16A11, RPEL1, RASL11B, ASGR1, SLC12A7, TTC28, POLR2A). Many loci have roles in cell cycle regulation or are associated with fibroid risk factors like blood pressure, BMI, and vitamin D levels. Loci were significantly enriched in DNA damage and cell cycle pathways. Of 588 significant predicted expression gene-tissue pairs, 173 unique genes were novel fibroid associations. These genes are also associated with cancers, estradiol, and endometriosis. Top enriched pathways included p53 signaling, HOTAIR, BRCA1DNA damage response, and pulmonary fibrosis signaling. In uterine tissue there were 15 novel GPGE associations. DISCUSSION/SIGNIFICANCE: Using this large and diverse data, we identified novel loci associated with fibroids that are enriched in hormone-response, DNA damage, and cell-cycle pathways. GPGE loci were in tumorigenesis and fibrosis pathways. These novel genetic loci and uterine gene expression findings may provide translational opportunities for novel fibroid treatments.

Public distrust in the US pandemic response has significantly hindered its effectiveness. In this community-based participatory research mixed-methods study, based on two datasets, we examined how distrust in COVID-19 vaccines relates to institutional distrust. We found that the Johnson & Johnson vaccine pause undermined trust in COVID-19 vaccines in general. Findings also suggest that vaccine distrust developed after participating in a study on COVID-19 testing. Increased distrust may be an unintended consequence of how healthcare and public health activities are presented and delivered, and research participation is structured. Both will continue without proactively addressing the root causes of distrust.