OBJECTIVES/SPECIFIC AIMS: There is currently a gap in the literature regarding the relationship between acute cellular rejection and frailty in LT patients. We aimed to evaluate the association between frailty and acute cellular rejection in LT patients. METHODS/STUDY POPULATION: Included were LT recipients from 2014-16 at a single center who had a frailty assessment prior to LT using the Liver Frailty index consisting of grip strength, chair stands, and balance. Frailty was defined by a Liver Frailty Index > 4.5. Data on acute cellular rejection at 3 months (primary outcome) and immunosuppression regimens were collected from medical chart review. Univariable and multivariable logistic regression assessed the associations between frailty and acute cellular rejection. RESULTS/ANTICIPATED RESULTS: A total of 241 LT recipients were included. Of these, 37% were female, 55% had Hepatitis C, and the median (IQR) age was 60 (54-65); 46 (19%) were classified as frail. 98% of patients were on a combination of mycophenolate, corticosteroids and tacrolimus on discharge compared to 80% by 3 months. Within the first 3 months post-LT, 7 (15%) of frail patients versus 10 (5%) (p = 0.02) of non-frail patients experienced acute cellular rejection. In univariable logistic regression, frailty was associated with a 3.3 times higher odds of acute cellular rejection at 3 months (95%CI 1.19, 9.26, p = 0.02); age (OR 0.91), Black race (OR 3.2), autoimmune disease (OR 2.3), and diabetes (OR 0.3) were also associated with acute cellular rejection at 3 months with a p-value<0.20. In a multivariate analysis, after adjusting for age, frailty remained significantly associated with rejection (OR 3.06, 95%CI 1.04, 9.01, p = 0.043). There were no significant differences in immunosuppression regimens or rates of mycophenolate dose reduction in the first 3 months between frail and non-frail patients. DISCUSSION/SIGNIFICANCE OF IMPACT: Frailty is associated with an increased rate of acute cellular rejection within 3 months post-LT, despite similar immunosuppression regimens and doses. Future studies should evaluate whether frailty should be considered in the management of immunosuppression in the early post-transplant period.

OBJECTIVES/GOALS: COPD is a progressive airways disease that results in death or disability. There is poor uptake of clinical guidelines (CPG) to manage COPD and studies to bridge this implementation gap have shown inconsistent results. Using implementation science principles we aim to understand COPD-CPG implementation determinants from providers’perspective. METHODS/STUDY POPULATION: The study is being conducted in ten VA Primary Care Clinics. Guided by the Consolidated Framework for Implementation Research (CFIR), a conceptual framework developed to guide systematic assessment of multilevel implementation contexts, we are using semi-structured guides to conduct key informant qualitative interviews (physicians, physician extenders and nurses), to support a formative evaluation. CFIR domains relevant to the study were determined by a multidisciplinary team. Informants are identified through online outreach and voluntary participation. Sampling adequacy will be assessed by achievement of code saturation. A qualitative template analysis will be used to summarize the barriers and facilitators of each component of COPD-CPG organized by CFIR-domain. RESULTS/ANTICIPATED RESULTS: We anticipate a list of modifiable and non-modifiable contextual, recipient (provider and patient), and COPD CPG content (innovation) barriers to implementation. Many settings do not have critical elements of these CPG, such as a standardized inhaler education/assessment pathway, patient education material, or pulmonary rehabilitation referral pathway. Existing literature indicate reasons behind the insufficient uptake of COPD CPG include low familiarity with guidelines, perception of minimal value of guidelines by physicians, and time constraints; we will present contextual, recipient and innovation determinants specific to our setting. DISCUSSION/SIGNIFICANCE: This comprehensive assessment of barriers and facilitators to COPD-CPG will inform tool development and implementation strategies identification to improve COPD CPG uptake. COPD is the most common veteran lung disease. Improvement in COPD care has enormous potential for benefit for local veterans, as well as potential for wider dissemination.

ABSTRACT IMPACT: Better understanding of the factors impacting disease severity and immunological response of MS patients on disease modifying therapy will enable better recommendations for vaccination options and risk mitigation strategies OBJECTIVES/GOALS: The Coronavirus Disease 2019 (COVID-19) and global health crisis has raised health concerns for patients with multiple sclerosis (MS). We aim to study the clinical characteristics, immunological laboratory data, and immunoglobulin response in patients with MS and COVID-19, to identify factors impacting disease severity and immune response. METHODS/STUDY POPULATION: Database search was done using DataDirect to search for MS patients who had tested positive for COVID-19 at the University of Michigan hospital. Patients with a positive nasopharyngeal swab polymerase chain reaction (PCR) for COVID-19 between March 1 and September 2020 were included. The primary outcome was the immunological laboratory data and immunoglobulin levels and the secondary outcome was their disease severity. We collected demographics, neurological history, MS treatment, Expanded Disability Scale Score (EDSS), comorbidities, and COVID-19 characteristics. A 7-point ordinal scale previously used to assess disease severity was used. Univariate and multivariate analyses will be performed to assess relationships between the collected variables. RESULTS/ANTICIPATED RESULTS: A total of 17 patients, mean age 53 (SD 11.6) years, mean disease duration, 6.2(SD 4.1) years were analyzed. 41% of patients had relapsing remitting multiple sclerosis, 17% had primary progressive MS. (88%) patients were on Disease Modifying Therapy (DMT) at the time of COVID-19 diagnosis. 2 patients died from COVID-19 complications. There was a higher proportion of patients with higher disease severity receiving Ocrelizumab. Only one patient showed positive IgG to SARS-CoV-2 after the resolution of infection. CBC with differential was obtained and a longitudinal follow-up of labs will be done. Regression analysis will be done to check the association between the use of DMT, immunological response, and COVID disease severity in them. The impact of COVID-19 on MS relapse, EDSS, and MRI activities will also be studied. DISCUSSION/SIGNIFICANCE OF FINDINGS: Recommendations to continue current DMT have been made, however, the immune response has not been correlated with the individual’s risk profile. Certain therapies may interfere with mounting a protective immune response of COVID-19 and this knowledge is crucial when advising patients regarding the choice of vaccine and risk mitigation strategies.

OBJECTIVES/GOALS: Lung transplant (LTx) candidates benefit from use of non-ideal donor organs. Each organ procurement organization (OPO) defines “acceptable” donor organs introducing unmeasured variation in donor pursuit. We characterized non-ideal donor pursuit among OPOs to identify drivers of risk aversion in LTx. METHODS/STUDY POPULATION: We queried the UNOS registry for adult donors who donated ≥1 organ for transplantation from 12/2007-12/2018. Non-ideal donors were those with any of age>50, smoking history ≥20 pack-years, PaO2/FiO2 (P/F) ratio<350, donation after cardiac death (DCD) status, or CDC increased risk (IRD) status. Non-ideal donor pursuit rate was defined as the proportion of non-ideal donors at each OPO from whom consent for lung donation was requested with lower numbers indicating increased risk aversion. We estimated the correlation between non-ideal and overall donor pursuit using a Spearman correlation coefficient. Adjusted non-ideal donor pursuit rates were estimated using multivariable logistic regression. RESULTS/ANTICIPATED RESULTS: Overall, 18,333 deceased donors were included and classified as ideal or non-ideal. Among 58 OPOs, rates of non-ideal donor pursuit ranged from 0.24-1.00 Figure). Of 5 non-ideal characteristics, DCD and IRD status were associated with the most and least risk aversion, respectively. Non-ideal donor pursuit was strongly correlated with overall donor pursuit (r = 0.99). On adjusted analysis, older age (OR 0.15, 95% CI 0.13-0.16), smoking history (OR 0.38, 95% CI 0.34-0.44), low P/F ratio (OR 0.12, 95% CI 0.11-0.14), and DCD status (OR 0.04, 95% CI 0.03-0.04) were all independently associated with significant risk aversion, corresponding to decreased rates of donor pursuit. DISCUSSION/SIGNIFICANCE OF IMPACT: OPOs differ in their levels of risk aversion in LTx and risk aversion is not uniform across selected categories of non-ideal lung donor. Consideration of new OPO performance metrics that encourage the pursuit of non-ideal lung donors is warranted.

Objectives/Goals: Cognitive decline is a known sequalae of intracranial radiation in the treatment of brain metastases. In this study, we investigate global structural changes in the brain akin to accelerated aging and compare aging kinetics between patients treated with whole-brain radiation therapy (WBRT) and stereotactic radiosurgery (SRS). Methods/Study Population: This retrospective study consists of patients with brain metastases treated with WBRT and SRS at our institution. Brain MRI images collected prior to radiation therapy and at approximately three and six months following radiation will be analyzed, excluding patients with evidence of worsening disease burden in the brain. Surface morphology of the cerebral cortex and sub-cortical structures will be extracted using Freesurfer and converted to graphs. Data will then be input into a validated graph convolutional neural network model to estimate brain age at each time point. A generalized linear model will be used to estimate the aging pace between baseline and follow-up for each subject within the whole brain as well as the sub-cortical structures, which will be compared between WBRT and SRS treatment groups. Results/Anticipated Results: We anticipate that intracranial radiation will accelerate brain aging to a greater extent following WBRT compared to SRS. Additionally, this accelerated aging will occur globally in the whole brain as well as within individual substructures, including the cerebral cortex, nucleus accumbens, amygdala, caudate, hippocampus, pallidum, putamen, and thalamus. Discussion/Significance of Impact: This study will demonstrate structural changes in the brain analogous to accelerated aging, supporting its potential use as an imaging biomarker to monitor cognitive decline after radiation therapy. Future work will explore the relationship between structural brain aging and assessments of neurocognitive function.

Objectives/Goals: Neuroendocrine malignancies are heterogeneous cancers with varied clinical outcomes, yet the molecular landscape driving this heterogeneity has not been fully characterized. Here, we investigate the gene expression and mutational profiles of neuroendocrine malignancies to better understand the underlying biology and therapeutic targets. Methods/Study Population: Patients with neuroendocrine tumors (NETs) and neuroendocrine carcinomas (NECs) treated at Cleveland Clinic (2000–2022) with molecular profiling (n = 66) were identified. Mutational and gene expression profiles were abstracted from electronic health records (EHR). Clinico-pathological characteristics and overall survival (OS) were obtained from EHR. Statistical analyses were performed by R v.4.0.5 and R package Limma for differential gene expression, as well as Chi-square, Fisher’s exact, and Wilcoxon rank sum tests. Results/Anticipated Results: The cohort consisted of 38 cases with NEC, 18 NET g3, and 10 NET g1/2. EZH2 and cyclin E1 were differentially over-expressed in NEC vs. NET (p < 0.05), while PTEN and MSLN were differentially under-expressed in NEC vs. NET (p < 0.005). Several recurrent alterations co-segregated with aggressive histology (NEC vs. NET): TP53 (p 60. Also, there was no difference in gene expression profiles between the two age groups among NETs or NECs. Discussion/Significance of Impact: This study explores the molecular landscape of NETs and NECs, revealing distinct gene expression and mutation profiles related to clinical outcomes. High expressions of cyclin D1 and EGFR were significantly associated with improved 2-year OS in NECs, highlighting potential therapeutic targets. Future studies are needed to validate these findings.

Objectives/Goals: This study evaluates the correlation between coronary artery calcium (CAC) scores and arteriosclerotic disease progression, determining if CAC scoring can predict high-risk individuals and guide early interventions to improve cardiovascular outcomes and disease prevention. Methods/Study Population: The retrospective descriptive study analyzed 800 coronary calcium scoring (CCS) reports to evaluate patterns of CAC scores and their relationship to arteriosclerotic disease progression. Data were collected from the Hospital Cardiovascular de Puerto Rico, focusing on CCS reports generated in 2024. The study population included patients aged 21 years and older who underwent CAC scoring during 2024. Instruments included radiology reports containing CAC scores and medical records detailing patient demographics, clinical history, and outcomes. Descriptive and correlation analyses were performed to explore associations between CAC scores and arteriosclerotic disease. Results/Anticipated Results: Preliminary findings from 800 CCS reports revealed that over half of the participants presented measurable coronary calcification, mainly among those aged 45–84 years with hypertension, diabetes, or hyperlipidemia. These results highlight the importance of CAC scoring as a noninvasive, cost-effective tool for early detection of subclinical atherosclerosis, enabling timely intervention and improved cardiovascular risk assessment. The study underscores its value in guiding prevention strategies and reducing disease burden. Discussion/Significance of Impact: This study highlights the calcium score as a precise, noninvasive tool for early detection of cardiovascular disease. It improves risk prediction and guides timely, targeted interventions to prevent complications and support effective, evidence-based cardiovascular prevention strategies.

OBJECTIVES/GOALS: Dyslexia is a common Reading Disability (RD) affecting 7-12% of the population and is associated with less cortical thickness (CT) in bilateral brain regions. However, the interaction between RD and a bilingual experience on CT is unknown, even though bilingualism is also associated with altered CT. METHODS/STUDY POPULATION: We studied 48 Bilinguals assigned to the Typical Reader group based on Oral Reading Recognition Test (ORRT) scores above 90 (avg=107 ± 14), 47 Bilinguals assigned to the RD group based on ORRT scores below 85 (avg=77 ± 5), 45 English Monolingual Typical Readers with ORRT scores above 90 (avg=102 ± 13) and 47 Monolinguals with RD based on ORRT scores below 85 (avg=78 ± 5). Participants (all from the Adolescent Brain & Cognitive Development Study) were 11.9 ± 0.7 years of age and the 4 groups were matched for sex, self-ratings of English, nonverbal reasoning, and combined household income. Structural magnetic resonance images were analyzed using CAT12 and all four groups were entered into a factorial analysis. RESULTS/ANTICIPATED RESULTS: Surprisingly, the main effect of Reading Ability did not reveal any regions where RD manifested less CT than Controls (raising the possibility that the findings from the only two prior reports were due to small samples). The main effect of Language Background revealed less CT in bilinguals in bilateral perisylvian regions (inferior frontal gyri, superior temporal gyri, and left Heschl's gyrus) consistent with prior reports. There was no interaction of Reading Ability by Language Background. Taken together, we found no differences in CT in those with RD relative to Typical readers and no evidence that the dual language experience affected this result in any way. DISCUSSION/SIGNIFICANCE: The lack of interaction between Reading Ability and Language Background indicates that a dual-language experience does not affect CT differently in those with RD and reduces concerns that RD in those who are bilingual needs to be given separate consideration in studies of CT neuroanatomy.

OBJECTIVES/GOALS: Tubulointerstitial damage in lupus nephritis (LN) is a strong predictor of progression to chronic kidney disease and end stage renal disease (ESRD). While complement activation mediates glomerular injury, the role of complement in renal tubular damage has not been evaluated. We investigated the association between complement activation and tubulointerstitial fibrosis. METHODS/STUDY POPULATION: Patients with LN were selected randomly between July 2014 - July 2016. Chromogenic immunohistochemistry was performed on formalin-fixed, paraffin-embedded, 4-µm human renal biopsy sections using unconjugated, murine anti-human Complement C9 (Hycult Biotech, clone X197) as a marker of the terminal complement activation. Positive control is C3 glomerulopathy and negative control is normal kidney. Tubular basement membrane C9 staining intensity were analyzed on semiquantitative scale 0 to 3 by a renal pathologist. Interstitial fibrosis/tubular atrophy were categorized into low (0–10%), medium (11–20%), or high (≥21%). Clinical parameters were assessed at time of biopsy and 6 months post biopsy. Bivariate associations were assessed between presence of tubular C9 (C9+) and other covariates. RESULTS/ANTICIPATED RESULTS: Renal biopsies from 30 LN studied, 23 (77%) of which had proliferative LN. There were 24 (80%) women, mean (SD) age 33 (12) years. Positive tubular C9 staining was observed in 7/30 (23%) biopsies. At time of renal biopsy, C9+ patients had significantly higher urine protein, compared to C9- patients: median (IQR) 6.2g (3.3-13.1) vs. 2.4g (1.3-4.6), p<0.01. The differences persisted at 6 months after induction therapy: 1.08g (1.0-8.3) in C9+ vs. 0.68g (0.2-2.1) in C9- patients, p = 0.06. There was no significant difference in creatinine at renal biopsy between the two groups. Tubular C9 deposition was associated with interstitial fibrosis: 49% had severe interstitial fibrosis vs. none in the C9- group, p = <0.01. Higher proportion of C9+ patients had moderate NIH Chronicity index: 42.9% vs 8.7% in the C9- group, p = 0.07. DISCUSSION/SIGNIFICANCE OF IMPACT: Tubular C9 deposition is significantly associated with proteinuria, interstitial fibrosis and increased chronicity which predict progression to ESRD and high mortality. This finding suggests that complement activation in the tubules may be linked to proteinuria and contribute to mechanism in tubulointerstitial damage in LN.

OBJECTIVES/GOALS: Assess molecular and cellular mechanisms of allograft loss in kidney biopsies using digital spatial profiling and clinical outcomes data. METHODS/STUDY POPULATION: Patients with chronic allograft dysfunction (CGD), enrolled in the Deterioration of Kidney Allograft Function (DeKAF) study, with or without eventual allograft loss, were included. CGD was defined as a >25% increase in creatinine over 3 months relative to a baseline. Kidney biopsy tissue was assessed by Nanostring GeoMX digital spatial profiling (DSP) after staining with anti-pan-cytokeratin, anti-CD45, anti-CD68, Syto-13, to identify specific cell populations, and Nanostring’s Whole Transcriptome Atlas (WTA), to quantify the distribution of transcripts across the biopsy. Up to 14 regions of interest (ROIs) were selected, with or without glomerulus. CIBERSORT was used to perform cell deconvolution. Clinical and outcomes data were from the DeKAF study and United States Renal Data System. RESULTS/ANTICIPATED RESULTS: Macrophage (M1) cell population abundance was significantly different in ROIs with glomerulus between graft loss and no graft loss. Principle component analysis of differentially expressed genes resulted in transcriptomes in ROIs that cluster together by clinical outcome of graft loss or no graft loss. There were 203 DEGs in ROIs with glomerulus that were different by graft loss or no graft loss. By pathway analysis, these 203 DEGS were enriched in the T-cell activation, integrin signaling and inflammation pathways. DISCUSSION/SIGNIFICANCE: DSP of kidney allograft biopsies allows for the identification and quantification of specific cell types, such as macrophages and molecular transcripts as potential drug targets. This data can be used to understand mechanisms of kidney allograft loss and may lead to improved immune suppression in kidney transplant recipients.

OBJECTIVES/GOALS: Untargeted metabolomics platforms are powerful biomarker discovery tools. However, the absence of uniform study design, data analysis, and reporting standards limits translation of this research into the clinical lab. The goal was to critically appraise existing untargeted metabolomics platforms that analyzed inborn errors of metabolism. METHODS/STUDY POPULATION: A search strategy was conducted in MEDLINE via PubMed from January 16, 2013, to January 16, 2023. The search strategy was limited to primary literature articles written in English that evaluated human subjects with inborn errors of metabolism (IEMs). Articles that performed targeted metabolomic analysis or analyzed non-human samples were excluded. Information on patient cohorts analyzed, sample types, and method design were extracted using a template. Categorical data are summarized as frequencies and percentages. RESULTS/ANTICIPATED RESULTS: A total of 96 distinct IEMs were evaluated by the different untargeted metabolomics methods included in this review. However, most IEMs (55/96, 57%) were evaluated by a single platform, in a single study, with a limited cohort size. Only one study validated their results using a separate, validation cohort. There was considerable diversity in the separation techniques and mass spectrometry instrumentation used by the studies to create their untargeted metabolomics methods. Slightly over half (59%) of the studies identified at least some of the metabolites detected in their samples with the highest level of confidence. Importantly, most of the included studies reported adherence to quality metrics, including use of quality control material (65%) and internal standards in their analysis (61%). DISCUSSION/SIGNIFICANCE: Future studies analyzing IEM patient samples with untargeted metabolomics platforms should progress beyond single-subject studies and evaluate the reproducibility of the research using a prospective, or validation cohort as well as confirm metabolite annotations with reference metabolites standards to generate clinically useful data.

Objectives/Goals: Endometrial cancer (ECa) incidence is rising rapidly in women under 40 years of age, and response rates and progression-free survival remain poor in advanced ECa. New targeted therapies are urgently needed. The objective of this study is to develop and test a potent inhibitor that targets midkine (MDK) for treating ECa. Methods/Study Population: Using the three-dimensional structure of human MDK, we rationally designed a small organic molecule (HBS-101) that directly binds with MDK and that functions as a MDK inhibitor. Microscale thermophoresis assay (MST) and in silico docking studies were used to demonstrate direct binding of HBS-101 to MDK. In vitro activity was tested using MTT, clonogenicity, and apoptosis assays. Mechanistic studies were conducted using Western blot, RT-qPCR, STAT3 and NFkB reporter gene assays, RNseq, and TEM. Status of MDK in ECa was determined using TNMplot database. Pharmacokinetics, Maximum Tolerable Dose, and in vivo efficacy studies were conducted using mouse models. HBS-101 efficacy in chemotherapy-resistant ECa was demonstrated using therapy-resistant primary ECa (TR-ECa) cells generated in-house. Results/Anticipated Results: MDK is highly expressed in ECa tumors compared to normal tissues. MDK and its receptors are expressed in established and primary ECa cell lines. Knockdown of MDK reduced ECa cell proliferation and colony formation. HBS-101 directly binds to MDK with nanomolar affinity, and a potential interaction surface on MDK was identified. HBS-101 decreased ECa cell viability and colony formation with an IC₅₀ ranging from 0.5 to 5 µM. Mechanistic studies showed that HBS-101 suppressed STAT3 and NFκB activity and induced cell death, ER stress, and ferroptosis. TR-ECa cells exhibited sensitivity to HBS-101. HBS-101 possesses favorable drug-like properties, including oral bioavailability, in vivo stability and little in vivo toxicity up to 10 mg/kg. HBS-101 significantly inhibited tumor growth in ECa PDX models. Discussion/Significance of Impact: HBS-101 is a novel therapeutic agent that targets MDK via a unique mechanism, shows favorable pharmacokinetics, and induces apoptosis and ferroptosis in ECa cells by blocking MDK signaling. HBS-101 shows promise in treating high-grade and therapy-resistant ECa and may help overcome the current lack of good treatment options for these patients.

OBJECTIVES/GOALS: Social isolation/loneliness is a public health crisis and one that is unlikely to be solved through pharmacology. Nonpharmacological approaches, such as dance, are needed. The objective of this study is to investigate the physiological correlates of dance-induced improvements in social connection. METHODS/STUDY POPULATION: Participants were randomly assigned to participate for 4 weeks (2 times per week, 90-minute sessions) in either 1) improvisational dance training (experimental group; n=7); or a 2) dance movie watching experience (control group; n=7). Before and after the intervention, using mobile brain-body imaging techniques, participants and their instructor had their brain (via electroencephalography) and body physiology (via photoplethysmography) recorded during a series of verbal and nonverbal interactive experiences. Participants were also video recorded via 4 surrounding cameras for later motion capture analysis. Neuropsychological assessments were also conducted before and after the intervention. RESULTS/ANTICIPATED RESULTS: We found that dance significantly increased social skills including empathy, interpersonal skills, emotional regulation, mindfulness, and attention. Additionally, we found that dance significantly increased interbrain synchrony during nonverbal experiences including theta (4-8 Hz), beta (12-35 Hz), and gamma (35-45 Hz) frequencies in the occipital lobe. Increases in interbrain synchrony were also positively correlated with increases in empathy. Additionally, intercardiac synchrony between the participant and instructor showed a significant correlation at post-intervention only. Future investigations will focus on the relationship between interbrain, intercardiac, and movement synchrony. DISCUSSION/SIGNIFICANCE: Our findings support the idea that dance increases interpersonal synchrony at the level of the brain, heart, and behavior. Understanding the neural and somatic mechanisms of social behaviors will help promote understanding and development of interventions for the critical problem of social isolation and loneliness.

OBJECTIVES/SPECIFIC AIMS: We sought to solicit and synthesize stakeholders’ ideas for how the Advance-CTR program can best increase the quality and quality of clinical and translational research in Rhode Island, and to apply these findings to address barriers and strengthen research capabilities across our partner institutions. METHODS/STUDY POPULATION: We utilized a Group Concept Mapping approach, involving university and Institution-based researchers and administrators. The process was conducted using the web-based concept mapping application CS Global Max (Concept Systems, Inc). Respondents were asked to provide their best ideas for promoting clinical and translational research in RI. These ideas were then organized by our project team into a set of unique items for consideration by attendees of an Advance-CTR retreat. Participants were tasked with sorting these ideas by theme (cluster), and were also asked to rate each idea according its importance and feasibility. Using the online software, these clusters and ratings were analyzed to identify key themes and to explore differences among sub-groups. RESULTS/ANTICIPATED RESULTS: The Group Concept Mapping exercise yielded 150 statements that were edited down to 78 unique ideas, and clustered into nine themes (e.g., institutional collaboration, training). Fifty-seven retreat participants completed the sorting and rating tasks of the concept mapping exercise. Overall, ideas rated as highly important and highly feasible included “providing seed grants to encourage new collaborations across basic science,” and “connecting researchers with common interests.” Top rated items varied across institutions and according to respondent demographics, allowing us to consider the unique issues relevant to particular groups. Relative rankings of clusters across groups revealed notable differences, such as higher importance placed on community engagement among administrators as compared with researchers, and differences in needs for internal support for research between universities. DISCUSSION/SIGNIFICANCE OF IMPACT: Group Concept Mapping was an effective and insightful participatory approach to engage our program’s stakeholders in developing ideas and identifying challenges to enhancing clinical and translational research in Rhode Island. Our results have implications for project decision-making and initiatives to facilitate translational research in RI. Thus, results have been presented to the Advance-CTR community via webinar, as well as Advance-CTR project leadership and advisory committees.

OBJECTIVES/GOALS: Familial Hemophagocytic Lymphohistiocytosis (FHL) is a systemic inflammatory disease, causing acute liver failure (ALF). Elevated Interferon gamma (IFN-γ) results in increased hepatic transcription of the chemokines CXCL9 and CXCL10. Inhibition of their receptor CXCR3 may reduce leukocyte recruitment and ameliorate hepatitis. METHODS/STUDY POPULATION: To determine the functional role of the IFN-γ-induced ligands, CXCL9 and CXCL10, in hepatic leukocyte recruitment via CXCR3 we used a prf-/- mouse infected with Lymphocytic Choriomeningitis Virus (LCMV) in our well-established model mimicking human FHL. We used AMG487, a small molecule CXCR3 antagonist, while maintaining intact IFN-γ signaling. Mice were sacrificed 10 days after infection when mice developed features of FHL: cytopenias, organomegaly, elevated serum ferritin and sCD25, and hepatic inflammation. Hepatic inflammation was characterized using flow cytometry, liver histology and noninvasive markers of hepatitis (ALT, liver size). RESULTS/ANTICIPATED RESULTS: AMG487 did not ameliorate the overall disease phenotype with mice developing similar FHL characteristics compared to control, including weight loss, elevation of ALT and sIL-2r as well as degree of thrombocytopenia and anemia. There was significant reduction of recruitment of CXCR3+CD4+ T cells and B cells in mice treated with AMG487. This indicates the importance of CXCR3 receptor in humoral response in FHL hepatitis. In addition, treatment with AMG487 resulted in reduction of CXCR3 expression in hepatic inflammatory monocyte (iMonos) measured by mean fluorescence intensity (MFI). DISCUSSION/SIGNIFICANCE: This is the first pre-clinical experience using AMG487, a small molecule CXCR3 antagonist, to treat FHL hepatitis. AMG487 changed the hepatic inflammatory milieu, reducing CD4 T-cell and B-cell recruitment, as well as CXCR3 expression on iMonos. However, it did not ameliorate FHL hepatitis and other therapeutic approaches should be pursued.

Objectives/Goals: Cerebral amyloid angiopathy (CAA) characterized by the accumulation of amyloid-beta in the cerebrovasculature, affects blood vessel integrity leading to brain hemorrhages and an accelerated cognitive decline in Alzheimer’s disease patients. In this study, we are conducting a genome-wide association study to identify genetic risk factors for CAA. Methods/Study Population: We genotyped 1253 additional AD cases using and curated existing genome-wide genotype data from 110 AD and 502 non-AD donors from the Mayo Clinic Brain Bank. We performed QC and imputation of all datasets. We conducted GWAS in AD only (N =  1,363), non-AD only, as well as the combined cohort (N = 1,865) by testing imputed variant dosages for association with square root transformed CAA using linear regression, adjusting for relevant covariates. To assess associations in the context of major CAA risk factors, we performed interaction analysis with APOEe4 presence and sex; and pursued stratified analyses. We collected peripheral gene expression measures using RNA isolated from 188 PAXgene tube samples of 95 donors collected across multiple time points. More than 1/3 of these participants have MRI measures collected. Results/Anticipated Results: Variants at the APOE locus were identified as the most significant in our study. In addition, several other variants with suggestive association were found under the main model adjusting for AD neuropathology (Braak and Thal). LINC-PINT splice variant remained associated with lower CAA scores in AD cases without the APOEe4 risk allele. To enhance the robustness of our findings, we are pursuing further expansion of our study cohort. In the periphery, we expect to identify expression changes associated with neuroimaging indicators of CAA and determine if variants and genes discovered via GWAS are implicated in these changes. Discussion/Significance of Impact: We expect this study will provide further insight into the genetic architecture underlying risk for CAA both in the context of significant AD pathology and without. Characterization of genetic variants and functional outcomes in the context of neuropathology may lead to new avenues of research aimed at identifying biomarkers and therapies to treat CAA

ABSTRACT IMPACT: This is the first examination of risk factors for prolonged opioid use after an ICU stay and will inform efforts to strengthen prescribing guidelines and care transition models for patients after critical illness. OBJECTIVES/GOALS: The majority of patients in intensive care units (ICU) receive opioids during admission, and up to 25% receive a prescription at discharge. However, transitions of care and prolonged use after discharge remain unknown. We aim to characterize risk factors for prolonged opioid use after an ICU stay. METHODS/STUDY POPULATION: A retrospective study using insurance claims from Optum Clinformatics ®Data Mart was conducted for opioid-naive adult patients (18-64 years) with an ICU admission from 2010 to 2019. The primary outcome was new persistent opioid use, defined as a continued prescription fill 91-180 days after discharge, in addition to a fill in the first 90 days. The primary exposure was an opioid fill at discharge. The ICU admission was characterized using the Clinical Classification System from the Agency of Healthcare Research and Quality, based on patients’primary diagnosis code. Diagnoses were combined into 11 groups highlighting the affected organ system/mechanism of injury. Logistic regression evaluated the associations of patient demographic and clinical characteristics with the probability of persistent opioid use. RESULTS/ANTICIPATED RESULTS: In this cohort of 90,721 patients discharged from the ICU, 3.3% continued to fill opioids at 6 months. An opioid prescription fill (OR 3.1; 95% CI 28 - 3.3) and benzodiazepine prescription fill (OR 1.6; 95% CI 1.4 - 1.8) within 3 days of ICU discharge were each significantly associated with the development of new persistent opioid use. Patient diagnosis groups of Musculoskeletal/Trauma (OR 2.3; 95% CI 2.0 - 2.6), Neoplasms (OR 1.6; 95% CI 1.5 - 1.9), and GI/Hepatobiliary (OR 1.5; 95% CI 1.3 - 1.8) were significantly more likely to develop new persistent use when compared to the Cardiovascular diagnosis group. DISCUSSION/SIGNIFICANCE OF FINDINGS: Opioid prescriptions at discharge after an ICU stay increase the odds of prolonged opioid use. These results will inform efforts to strengthen prescribing guidelines and care models after a critical illness. Further work will characterize the trajectory of prescribing and patient exposure to high-risk prescribing after ICU discharge.

OBJECTIVES/GOALS: This study examines sex differences in brain structure in youths with disruptive behavior disorders (DBD). We use measures of gray matter volume (GMV) in regions-of-interest implicated in the pathophysiology of conduct problems and a whole-brain analysis of cortical thickness. We also examine unique associations between brain structure and callous-unemotional traits. METHODS/STUDY POPULATION: This study included 90 children with a DBD (30 females) aged 8-16 and 50 Healthy Controls (20 females) matched for age and IQ. Children received a diagnostic evaluation using the K-SADS. Pre-processing was conducted using FreeSurfer. For ROI analyses, multivariate GLM models were conducted in SPSS for estimates of GMV to examine the main effects of diagnosis and sex, and sex-by-diagnosis interactions. Whole-brain analyses were conducted in FreeSurfer. Associations were examined between structure and parent ratings of callous-unemotional (CU) traits using the Inventory of Callous-Unemotional Traits in regression analyses in the DBD group, while controlling for the variance in aggressive behavior using the Child Behavior Checklist Aggressive Behavior Scale. All analyses controlled for differences in intracranial volume. RESULTS/ANTICIPATED RESULTS: Relative to controls, children with DBD showed reduced GMV in the bilateral amygdala (left: p = .004; right: p = .04). Sex-by-diagnosis interactions were observed in the left ventromedial prefrontal cortex (p = .004), right insula (p = .001), right inferior frontal gyrus (p = .02), and bilateral anterior cingulate (left: p = .02; right: p = .01) in which DBD males showed lower and DBD females showed higher GMV relative to respective controls. For whole-brain analyses, a significant sex-by-diagnosis interaction was observed in the left ventromedial prefrontal cortex and supramarginal gyrus indicating that DBD males showed lower and DBD females showed higher cortical thickness relative to respective controls. Sex-by-CU traits interactions were observed for left amygdala and ACC volumes. DISCUSSION/SIGNIFICANCE OF IMPACT: The current study provides evidence of reduced amygdala volume in children with DBD, and interactions between sex and diagnosis in the ventromedial prefrontal cortex and supramarginal gyrus, which may have implications for identifying sex-sensitive neural biomarkers. CONFLICT OF INTEREST DESCRIPTION: Disclosures: Dr. Sukhodolsky receives royalties from Guilford Press for a treatment manual on CBT for anger and aggression in children. Drs. Ibrahim, He, Pelphrey, McCarthy, and Mr. Li have no biomedical financial interests or potential conflicts of interest to declare related to this present study.

Clinical and Translational Science Awards (CTSAs) produce educational content spanning the continuum of clinical and translational research. However, the way in which this content is organized should be standardized. We describe our methodology and process for re-organizing the content in our hub’s Online Educational Library (OEL) using eight clinical research professional competency domains. We also analyze data from our OEL on viewership and related factors. Users were more likely to watch videos that were newer and hosted on YouTube. The public availability of competency categories had mixed effects on viewership. We discuss the need to refine these categories to improve video viewership in CTSA OELs.