Objectives/Goals: We have shown that parathyroid hormone-related protein (PTHrP) is enriched at the LIFR promoter in breast cancer cells and inhibits the expression of dormancy-associated genes including LIFR. The objective of this study is to define where all PTHrP binds DNA and identify pathways that are regulated by PTHrP that promote breast cancer colonization of the bone. Methods/Study Population: In this study, we use human estrogen receptor-positive MCF7 breast cancer cells which we and others have reported lie dormant in the bone. MCF7 cells were engineered to express either PTHrP with an HA-tag (MCF7P), or a vector control (MCF7V). We use Cleavage Under Targets and Release Using Nuclease (CUT&RUN), a method of mapping protein-DNA interactions, to define where PTHrP binds DNA. Here, an HA-specific antibody identifies regions of DNA that are bound to PTHrP in MCF7P cells compared to MCFV cells. Next, we perform DNA sequencing and gene set enrichment analysis (GSEA) on genes identified by CUT&RUN to identify pathways that are regulated by PTHrP. These experiments will determine how PTHrP regulates dormancy and breast cancer colonization in the bone. Results/Anticipated Results: We completed IgG (-control), H3K4me3 (+ control), and HA (PTHrP) CUT&RUN on MCF7V and MCF7P cells, and submitted DNA for sequencing. This study will define where PTHrP binds the genome and identify pathways regulated by PTHrP. Previously, through ChIP-qPCR we showed that PTHrP binds the LIFR promoter to repress LIFR expression. Given this result, we expect that PTHrP binds to the promoters of dormancy-associated genes including LIFR in MCF7P cells compared to MCF7V cells. PTHrP may be involved in regulating other processes besides dormancy to induce expansion of breast cancer cells in the bone, so we will use GSEA to identify pathways that are altered in MCF7P cells when PTHrP is over-expressed compared to MCF7V cells. Together, this will define how PTHrP regulates gene expression of bone metastatic breast cancer cells. Discussion/Significance of Impact: This study will unveil mechanisms of metastatic breast cancer expansion in the bone by defining where PTHrP binds the genome to regulate gene expression. These findings will reveal therapeutic vulnerabilities that will be used to target bone-disseminated tumor cells to prevent lethal recurrence.

Objectives/Goals: UW-Madison ICTR Collaborative Network (ICON) is a team of research navigators who provide wrap-around services for investigators (e.g. study and recruitment planning). In 2025, an ICON objectives was to consult with newly awarded investigators to expedite their study activities. Methods/Study Population: The ICON team assisted postdoctoral (n=3) and faculty (n=8) investigators awarded 12- or 18-month Institute for Clinical and Translational Research (ICTR) pilot awards beginning in May or July 2025. ICON addressed concerns related to regulatory documentation creation, recruitment and retention, study personnel training, document editing, etc. Ten of 11 investigators consulted with ICON shortly after award onset, with plans for check-ins at the 6- and 12-month mark of their grant. Following the initial consultation, ICON devised an individualized assistance plan for each investigator. Investigators completed multiple surveys regarding their opinion on the usefulness of the service. Results/Anticipated Results: The results of the first survey indicated that three investigators found the ICON service to be highly valuable (highest ranking), overall, the mean value was 4.1 on a 5-point Likert scale. The written feedback from the investigators indicated that the service would have been more useful earlier in the pilot award process, such as prior to submission. The ICON team continues to execute the individualized assistance plan for each investigator and will provide tailored assistance at the 6- and 12-month check-ins to meet the investigator’s needs. We will survey the investigators quarterly for feedback on the usefulness of the service. Discussion/Significance of Impact: Survey feedback indicates the value of the ICON service but suggests that for some the consultation was delayed. The ICON team will continue to check-in with this cohort to determine if further assistance is needed. In the future, ICON will meet with awardees earlier in the application process.

Every research study that includes volunteer participants requires safety assurances in proportion to the risks of the study. Investigator-initiated clinical research can present unique regulatory challenges particularly for studies with a risk profile that warrants more oversight than minimal risk but less than for large, commercial, or high-risk research. The use of an independent safety officer (ISO) offers a middle way of right-sizing oversight to match the risk. ISOs are clinicians or researchers with relevant expertise who are independent of the investigator and the research study. Their relationship to the study is defined by a formal charter which is aligned with the protocol and Data and Safety Monitoring Plan to address the oversight process, responsibilities of the ISO, and clearly describe the variables to be monitored. The ISO responsibilities include reviewing safety data, adverse events, recruitment, demographics, study progress, data quality, protocol changes, and any new scientific information that pertains to the trial. Finally, the ISO reports in their review on any significant findings may propose modifications to the study or a need to stop the trial.

Implementation science offers a compelling value proposition to translational science. As such, many translational science stakeholders are seeking to recruit, teach, and train an implementation science workforce. The type of workforce that will make implementation happen consists of both implementation researchers and practitioners, yet little guidance exists on how to train such a workforce. We—members of the Advancing Dissemination and Implementation Sciences in CTSAs Working Group—present the Teaching For Implementation Framework to address this gap. We describe the differences between implementation researchers and practitioners and demonstrate what and how to teach them individually and in co-learning opportunities. We briefly comment on educational infrastructures and resources that will be helpful in furthering this type of approach.

This article describes the Implementation Science (IS) Scholars Program at the University of Arkansas for Medical Sciences (UAMS). The program’s goal is to translate knowledge, approaches, and methods from IS to front-line clinicians in an academic medical center, thereby supporting its goals as a learning health system and promoting a dynamic workforce of IS-informed change leaders. Initiated in 2020, the program is relatively unique in that it attempts to translate concepts and knowledge from IS to clinicians to improve their skills as implementers and change agents. The program is supported by the Translational Research Institute, the UAMS’ awardee of the Clinical and Translational Science Award Program. The two-year program provides 20% salary coverage, bespoke didactics, and close mentoring on a Scholar-initiated project to improve care in their clinical context. The program has trained four cohorts of Scholars over the program’s initial five years. We describe the program, our evaluation of it thus far, and future plans. The program has contributed to numerous healthcare improvements and served as a gateway to future implementation and other research activities among some Scholars.

Twelve evidence-based profiles of roles across the Clinical and Translational Science (CTS) workforce and two patient profiles were developed by CTS Personas collaborators in 2019 as part of the CTSA Program National Center for Data to Health (CD2H). Based on feedback received from the community, CTS Personas team members collaborated to produce five additional Personas to broaden representation of the CTS workforce and enhance the existing portfolio. This paper presents the rationale and methodology used in the latest CTS Personas initiative. This work also includes an implementation scenario incorporating multiple Personas. Using the new National Institutes of Health’s (NIH) Data Management and Sharing Policy as an example, we demonstrate how administrators, researchers, support staff, and all CTS collaborators can use the Personas to respond to this new policy while considering the needs of service providers and users, CTS employees with short- and long-term needs, and interdisciplinary perspectives.

Objectives/Goals: The PSCTSI provides critical resources and services at PSU/PSCOM. Our objectives are to map PSCTSI resources among the PSU/PSCOM research infrastructure, increase researcher knowledge of resources, services, and processes, and ensure improved PSCTSI integration within the research infrastructure. Methods/Study Population: PSCTSI will utilize process and resource mapping techniques and systems analysis to align our services and resources within the PSU/PSCOM research infrastructure. Methods: Identify and map PSU/PSCOM research infrastructure processes, services, offices, and resources. Identify and map PSCTSI resources and services within the PSU/PSCOM infrastructure. Highlight areas where PSCTSI resources and services fill critical gaps in the PSU/PSCOM infrastructure Results/Anticipated Results: Navigating the research process is often confusing for researchers, especially where there are gaps within the infrastructure. Process and resource mapping techniques and systems analysis can provide a complete picture of PSCTSI’s vital role and impact on the PSU/PSCOM research infrastructure. Results of this analysis will benefit three audiences: PSU/PSCOM researchers – to better understand research processes, PSU/PSCOM research leadership – to illustrate the importance and synergy of PSCTSI within the research infrastructure, and PSCTSI and other CTSAs – to share their impact and ensure improved integration Discussion/Significance of Impact: CTSAs rely on external and potentially institutional support despite their essential roles within the research infrastructure. Resource and process mapping of PSCTSI within PSU/PSCOM’s infrastructure has two benefits: improving knowledge on resources and services and ensuring improved integration.

OBJECTIVES/GOALS: Hepatitis B and C virus causes inflammation of the liver and can lead to cirrhosis, liver failure, and hepatocellular carcinoma. The aim of this study is to generate a modeled estimate of changes in hepatitis B and C prevalence, and future sequelae, that accounts for recent mass migration to the European Union stemming from 50 high-emigration countries. METHODS/STUDY POPULATION: Total migrant population from 2013-2017 was obtained from the Eurostat population database. Demographics including country-of-origin, sex, and age distributions were used to determine migrant contributions to HBV and HCV prevalence where available. Undocumented migration estimates were obtained from the Institute of Migration database. Country-of-origin HBV and HCV prevalences were obtained for the select 50 country-of-origin nations from the Polaris Observatory and from systematic reviews. Disease progression was estimated using HBV and HCV outcome data for total populations from treatment guideline publication from the European Association for the Study of the Liver. RESULTS/ANTICIPATED RESULTS: Between 2013 and 2017, a total of 11,030,786 documented migrants born outside the EU arrived to the 30 nations. Germany, United Kingdom, and Spain received the greatest influx of persons and the majority of migration stemmed from countries in West Asia, the Middle East, and Africa. A significant proportion of total migration was driven by conflict-related crisis in Syria, and East and North Africa. The most significant increases in estimated total hepatitis case numbers, national prevalence increases, and future sequelae were seen in Germany and Sweden. DISCUSSION/SIGNIFICANCE OF IMPACT: Mass migration has significantly changed HBV and HCV disease burden in Europe over the past 5 years. Consequently, long-term outcomes of cirrhosis and HCC are also expected to increase. These increases are likely to disproportionally impact individuals of the migrant and refugee communities. HBV and HCV surveillance and management programs must strategically focus on individuals from high-burden age cohorts and nations. Screening and treatment would aid WHO elimination efforts while benefiting both the vulnerable individuals and host nations through reduction of morbidity, mortality, and associated healthcare expenses.

OBJECTIVES/SPECIFIC AIMS: Our primary objectives were to examine the impact of biologic cost sharing on 1) adherence to biologics and 2) persistence on biologics in inflammatory bowel disease (IBD) patients. Our secondary objective was to assess the effect of biologic cost sharing on clinical IBD outcomes, including rates of hospitalization, abdominal surgery, and corticosteroid treatment. METHODS/STUDY POPULATION: This retrospective cohort analysis used a national insurance claims database (Optum Clinformatics DataMart) to assess adult IBD patients enrolled in medium or large private insurance plans from 2007-2016. Patients were followed for one year of continuous enrollment after their index biologic claim. We assessed adherence to biologic medications (medication possession ratio >0.8) dependent on patient cost sharing, as measured by an employer-plan’s average out-of-pocket biologic medication cost. We also examined the effects of patient cost sharing for biologics on need for hospitalization, abdominal surgery, or corticosteroid treatment. We used multivariate logistic regression models adjusting for clinical and demographic characteristics. We estimated the effect of cost sharing on biologic therapy persistence using repeated measures proportional hazard survival models. RESULTS/ANTICIPATED RESULTS: We identified 2,193 adult IBD patients who initiated biologic therapy and met study criteria (Crohn’s disease 66.1% vs. ulcerative colitis 24.9%, mean age 40.8 years, mean Charlson index 0.50). Median [IQR] out-of-pocket cost per 30-day biologic prescription was $62 [$34 - $157]. 66.9% of patients were adherent to biologic therapy. Higher out-of-pocket costs for biologics were associated with increased odds of nonadherence; patients with ulcerative colitis were more price-responsive than patients with Crohn’s disease or indeterminate colitis (Figure 1). However, higher out-of-pocket biologic costs were not associated with increased odds of all-cause or IBD-related hospitalization, IBD-related surgery, or corticosteroid prescriptions for IBD flares. Patients whose out-of-pocket costs were less than $10 per 30-day biologic prescription persisted on biologic therapy for significantly longer than patients who paid >$10 (Figure 2). DISCUSSION/SIGNIFICANCE OF IMPACT: Nonadherence to biologics increases when IBD patients face higher out-of-pocket costs, particularly for ulcerative colitis patients. However, this is not associated with worse clinical outcomes. Patients with cost-sharing<$10 persisted on biologics longer than patients whose cost sharing exceeded $10.

Objectives/Goals: Our aim is to compare scores collected from a health utilities measure (Health Utility Index, HUI) to those collected from a profile measure (Child Health Ratings Inventories, CHRIs- Global) among youth with newly diagnosed, high-risk classic Hodgkin lymphoma. Methods/Study Population: We will analyze existing data collected during the Children’s Oncology Group AHOD 1331 trial, which was a phase 3 clinical trial comparing the efficacy of adding brentuximab vedotin to standard-of-care treatment with multiagent chemotherapy in children and adolescents with high-risk Hodgkin lymphoma. The study also had a prespecified patient-reported outcomes (PRO) secondary aim, which involved recruiting a subset of the initial 309 patients aged 11 years or older enrolled in the trial for serial PRO measures taken over the trial period. Health-related quality of life (HRQoL) was assessed by CHRIs, HUI version 2, and HUI version 3 assessments at six planned points throughout treatment. Results/Anticipated Results: The first step of our analysis will be to ascertain agreement in scoring for parent–child dyads for the HUI2, HUI3, and CHRIs scores by comparing mean scores via two-sample t-testing. Bland–Altman plots will be constructed to compare agreement between the scores for HUI2/3 and the CHRIs. Similarly, Spearman’s correlation coefficients will be calculated for CHRIs with HUI2/3 for both parents and children. We hypothesize the CHRIs and HUI scores should roughly correlate with one another, but there may be divergence of correlation because the HUI has greater emphasis on functionality (e.g., sensation, mobility), and the CHRIs further emphasize social and emotional well-being in addition to physical health. Discussion/Significance of Impact: The composite score of the HUI 2/3 has allowed for direct comparison with other global HRQoL measures, providing greater clarity of its performance in different patient populations and clinical settings. The current study will improve understanding of the HUI 2/3 performance in a pediatric cancer population over time.

OBJECTIVES/GOALS: Provider and hospital factors influence quality, but granular data is lacking to assess their impact on renal cancer surgery. The Maryland Health Service Cost Review Commission (HSCRC) is an independent state agency that promotes cost containment, access to care and accountability. Within HSCRC, we aimed to assess the impact of surgeon and hospital volume on 30-day outcomes after renal cancer surgery. METHODS/STUDY POPULATION: Data on renal surgery were abstracted from the Maryland HSCRC from 2000-2018. We excluded patients younger than 18, patients without a diagnosis of renal cancer, and patients concurrently receiving another major surgery. Volume categories were derived from the distribution of mean cases performed per year. We used adjusted multivariable logistic and linear regression models to identify associations of surgeon and hospital volume with the length of stay, days in intensive care, cost, 30-day mortality, readmission, and complications. RESULTS/ANTICIPATED RESULTS: A total of 10,590 surgeries, completed by 669 surgeons at 48 hospitals, met criteria. The 25th percentile for cases per year was 1, the 50th percentile was 1.2, and the 75th percentile was 2.6. After adjusting for patient factors and cumulative surgeon experience, high volume surgeons had the greatest decrease in length of stay (β: −1.65, P<0.001) and mortality risk (OR: 0.27, 95% CI: 0.10-0.71) compared to rare volume surgeons. Low volume surgeons had the greatest cost decrease (β: -$7,300, P<0.001) compared to rare volume surgeons. Medium volume hospitals had statistically lower average costs than rare volume hospitals (β: $−2,862, P = 0.005). There were no other clinically and statistically significant relationships between volume and measured outcomes. DISCUSSION/SIGNIFICANCE OF IMPACT: Almost half of the urologists studied performed an average of one renal cancer case per year. Greater surgeon volume was associated with shorter length of stay and decreased mortality risk. Hospital volume did not have a meaningful relationship to outcomes. Other factors such as tumor, surgeon, and hospital characteristics or case-mix may associate with outcomes and could be confounders.

OBJECTIVES/SPECIFIC AIMS: Intrahepatic Cholestasis of Pregnancy (ICP) is the most common liver disease unique to pregnancy. Progression of non-alcoholic fatty liver disease (NAFLD) has been linked to the dysregulation of bile acid homeostasis. However, an association of NAFLD with ICP has not been previously evaluated. We evaluated the association between ICP and NAFLD and associated metabolic risk factors, including obesity, dyslipidemia, hypertension, and diabetes. METHODS/STUDY POPULATION: A single-center, retrospective case-control study was conducted between January and December 2017 in a primarily Latina population in a New York City health system with a high prevalence of ICP, 2.53% of all pregnancies (compared to 0.32% nationally). Pearson’s chi-square or Fisher’s exact test and Wilcoxon rank-sum tests were performed to evaluate association of ICP with categorical variables and continuous variables, respectively. Unadjusted odds ratios (OR) with 95% confidence intervals (95% CI) were calculated in comparison with the control group for clinically significant outcomes. RESULTS/ANTICIPATED RESULTS: 149 pregnancies complicated by ICP were identified from electronic medical records; 200 controls were matched by delivery year. Hispanic women were more likely to be diagnosed with ICP than non-Hispanic women (OR 1.90, 95% CI 1.87-3.03). ICP and control patients were similar for: median age (OR 1.02, 95% CI.99-1.06), nulliparity (OR.79, 95% CI.48-1.30) and prevalence of hepatitis C (OR 1.35, 95% CI.08-21.67). In regards to metabolic risk factors, ICP patients and control patients were similar in prevalence of obesity (body mass index>30kg/m^2; OR 1.01, 95% CI.62-1.61), hemoglobin A1c>5.5% (OR.80, 95% CI.34-1.9), total cholesterol >200 mg/dL (OR 4.15, 95% CI.83-20.84), and prevalence of hypertension (OR.69, 95% CI.31-1.52). Median bile acid levels were 30.6 µmol/L (IQR 11.6, 32.7) in the ICP group. ICP patients had higher median alanine aminotransferase (ALT) (32 vs. 16 U/L, p<.0001), alkaline phosphatase (181 vs 128 U/L, p<.0001) and total bilirubin levels (0.5 vs 0.35, p<.0001) compared to controls. ICP patients were more likely to have ALT levels > 50 U/L (2 times the upper limit of normal; OR 3.22, 95% CI 1.48-7.03). ICP patients were significantly more likely to have a history of biliary disease (OR 3.29, 95% CI 1.39-7.80). ICP patients were more likely to have evidence of steatosis on liver imaging (OR 4.69, 95% CI 1.68-13.12) than non-ICP patients. When evaluating a diagnosis of NAFLD based on ICD-10 code or evidence of steatosis on liver imaging, ICP patients were significantly more likely to have a diagnosis of NAFLD than controls (OR 5.7, 95% CI 2.08-15.65). DISCUSSION/SIGNIFICANCE OF IMPACT: ICP appears to be associated with NAFLD independently of metabolic risk factors such as obesity, dyslipidemia, hypertension, and diabetes, suggesting a direct link between NAFLD and ICP. If findings are confirmed, ICP patients, especially those with elevated ALT, would benefit from screening for NAFLD and linkage to liver specialty care postpartum.

OBJECTIVES/GOALS: Maternal mental health, such as post-traumatic stress disorder (PTSD), is closely linked to child mental health. PTSD in mothers is associated with their children’s emotional responses. We examined associations between maternal PTSD and child brain function during emotion regulation. METHODS/STUDY POPULATION: Eight children ages 10-12 years, whose mothers had trauma histories, performed the Emotional N-Back task during functional MRI scanning. Mothers and children each reported on their trauma exposure and PTSD symptom severity. BOLD response to fearful faces during the Emotional N-Back was extracted from two specific brain regions of interest, amygdala and anterior cingulate cortex. These regions are involved in emotional response and attentional control, which are processes intrinsic to emotion regulation. An independent samples t-test was conducted on children’s BOLD response to fearful faces, with maternal PTSD symptom severity (high, low) as the independent variable. A parallel analysis was conducted with child PTSD symptom severity (high, low) as the independent variable. RESULTS/ANTICIPATED RESULTS: We found a main effect of maternal PTSD within brain regions of relevance to implicit emotion regulation. Compared to children whose mothers reported low PTSD symptom severity (n=4), children whose mothers reported high PTSD symptom severity (n=4) showed greater responsiveness to fearful faces in anterior cingulate cortex (t=2.04, p=.09,d=1.44) and amygdala (t=2.44, p=.05, d=1.72) at trending significance. A parallel analysis with child PTSD symptom severity showed no differences in brain function by this factor (ps=.55-.61). DISCUSSION/SIGNIFICANCE: Our pilot study is the first, to our knowledge, to examine associations between maternal PTSD and brain function during emotion regulation in their children. This study lays a foundation for future work; our goal is to explore dysfunction in emotion regulation neurocircuitry as one mechanism linking maternal PTSD to their children’s mental health.

Objectives/Goals: To use patient-level Center for Medicare and Medicaid Services (CMS) mandated quality metrics for inpatient rehabilitation facilities (IRFs) to develop and validate predictive models of functional recovery and interactions of baseline characteristics with therapy time. Methods/Study Population: Retrospective cohort study of a national US sample of ~40,000 adults with a primary diagnosis of stroke admitted to IRFs in 2023. Records will be randomly allocated to equal training and validation samples. We will use a random forest approach to generate predictive models for self-care and mobility functional outcomes using patient baseline and demographic data from a CMS-mandated assessment for IRFs (Section GG). We will also examine how predictive variables modulate the effects of occupational, physical, and speech-language therapy minutes. The random forest is a machine-learning approach that trains multiple models and combines their predictions to improve their overall performance. Results/Anticipated Results: Predictive models developed from the training sample will be applied to the validation sample to confirm their capacity to support new observations. Preliminary results will be reported, including the F1 score and area under the curve (AUC), with 95% confidence intervals. A unique feature of this study is the large sample, which contrasts with prior research in stroke rehabilitation using machine learning approaches. This study will produce powerful models that will inform the design of a clinical decision-support tool for application into clinical practice in a future study. Discussion/Significance of Impact: By using CMS-mandated quality metrics that are collected as part of standard clinical practice in IRFs, results will support clinical interpretation and application of metrics and inform the development of a clinician-facing intervention to support personalized rehabilitation approaches.

Objectives/Goals: To achieve the greatest impact, protocols must be rigorous and feasible. The University of Minnesota’s Clinical Research Support Center developed a feasibility review (FR) to identify barriers, offer solutions, and strengthen translation through multidisciplinary expertise. Improvements are tracked using an Impact Scoring System. Methods/Study Population: Established in 2018, the FR was designed to address operational barriers that delay study start-up and help researchers strengthen protocols before regulatory submission and implementation. Drafts are reviewed by a multidisciplinary panel with expertise in logistics, recruitment, biostatistics, monitoring, and regulation. Each protocol undergoes a 1-week review process culminating in a 90-minute structured discussion between the panel and study team. A written report summarizes strengths, concerns, available resources, and next steps. To assess impact, we developed a novel scoring system that assigns each protocol a pre- and post-review score, objectively measuring how protocols improve in response to feedback. Results/Anticipated Results: Since its launch, 186 clinical studies from nine schools and colleges have completed FR. Study teams report high satisfaction (average rating 4.8/5), noting clearer recruitment plans, improved logistics, and valuable resource connections. The Impact Scoring System provides measurable evidence of change: on average, protocols increased their score by 28% from pre- to post-review, reflecting greater operational readiness and likelihood of success. Tracking also shows that FR participation is associated with fewer protocol-related IRB stipulations, offering an additional objective indicator of improved quality. Together, these results demonstrate both the perceived value and the measurable operational impact of the FR process. Discussion/Significance of Impact: The FR addresses operational barriers early, ensuring protocols are both rigorous and feasible before launch. Its Impact Scoring System provides objective evidence of improvement and a scalable model other institutions can adopt to advance rigor, efficiency, and trustworthy translation.