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.

There is a general separation between medical and business education, despite the substantial intersection of these fields in hospital and industry settings. This separation hinders medical innovators, who rarely have opportunities to see how entrepreneurial concepts apply to the translation of medical innovation. To address this, the Office of Entrepreneurship at Mayo Clinic Florida provides experiential entrepreneurial training through an internal pitch competition: the alligator tank (AT). This study evaluates the impact and efficacy of the first seven years of the AT. Key metrics include a modified technology readiness level (TRL), determined by the presence of a disclosure, prototype, and preliminary data. A high TRL is not indicative of success in the pitch competition, leveling the playing field for early-stage inventions. The AT is characterized by broad representation across academic and professional ranks, departments, and genders, as well as strong participant retention. Innovators with prior disclosures are more likely to win the AT, presumably due to their experience. The AT serves as an institution-wide initiative that cultivates an inclusive, supportive innovation culture, fosters interdisciplinary medical innovation, and helps advance innovation maturity. Adopting the AT framework may help similar medical systems build an innovation ecosystem within their institutions.

Enrollment into a prospective cohort study of mother–preterm infant dyads during the COVID-19 pandemic progressed slower than anticipated. Enrollment occurred during the first week after preterm birth, while infants were still hospitalized. We hypothesized that slower enrollment was attributable to mothers testing positive for COVID-19 as hospital policies restricted them from entering the neonatal intensive care unit, thus reducing interactions with research staff. However, only 4.5% of 245 screened mothers tested COVID-19 positive. Only 24.9% of those screened, far fewer than anticipated, were eligible for enrollment. Assumptions about pandemic-related enrollment barriers were not substantiated in this pediatric cohort.

Extensive health inequities exist for persons with criminal-legal involvement in the USA. Researchers, both novice and experienced, are critical in documenting these inequities and implementing programs that address the many health and social problems of this population. However, working with currently or formerly incarcerated persons brings new challenges to researchers that may have not been previously considered as necessary. Because incarcerated persons were systemically exploited by biomedical researchers until reform following the Civil Rights Movement, resulting in their designation as a vulnerable population in the Code of Federal Regulations, enhanced protections are necessary in implementing contemporary research involving incarcerated persons. These enhanced protections can delay or prolong the regulatory approval process, particularly to the novice carceral system researcher, which may discourage some from engaging with this important population. Drawing on the many years of experience working with incarcerated persons accumulated by the Sexual Health Empowerment (S)HE Team at the University of Kansas Medical Center (KUMC), this article offers some concrete steps toward getting started in this work.

OBJECTIVES/GOALS: Intra-aortic balloon pumps are commonly used as circulatory support in patients with critically reduced cardiac function. The goal of this study is to estimate the incidence of mesenteric ischemia as an understudied vascular complication and to describe the clinical and radiographic characteristics of patients experiencing this complication. METHODS/STUDY POPULATION: We will be conducting a retrospective analysis of the electronic medical records of all patients who underwent intra-aortic balloon pump (IABP) placement between October 2020 and April 2023 at our academic medical center to identify the incidence of mesenteric ischemia. We will describe the clinical course of these patients and characterize them based on demographic features and risk factors for vascular complications including medical comorbidities. Finally, we will assess available chest x-ray and thoracoabdominal CT imaging for adequacy of balloon tip positioning, concordance between balloon size and aortic dimensions, and compromise of any visceral arteries in patients who experienced mesenteric ischemia vs. those who did not. RESULTS/ANTICIPATED RESULTS: We anticipate approximately 150 patients to have recieved IABPs over this period with at least 4 known cases of mesenteric ischemia. We will describe the clinical presentation of these cases and their often fatal outcomes. We expect several known risk factors will be present in these patients, including history of peripheral vascular disease, diabetes, or smoking history. On chest x-ray, we predict balloon tip positioning to be suboptimal (defined as >5cm below the aortic arch) in many patients, both those with mesenteric ischemia and those without, but a greater discordance on CT imaging between balloon size and aortic dimensions with greater evidence of visceral compromise in patients with mesenteric ischemia compared to those without. DISCUSSION/SIGNIFICANCE: Mesenteric ischemia is a serious and poorly studied complication of intra-aortic balloon pumps. Understanding the hospital course, clinical characteristics, and radiographic features present in these patients may guide clinicians in the early recognition and management of this potentially fatal complication.

ABSTRACT IMPACT: This work has the potential to help clinicians decide which infants exposed to in utero opioids, will need to be treated early or can be discharged home early based on their risk, thus reducing prolonged hospitalization OBJECTIVES/GOALS: To develop and validate a prediction model with inclusion of clinical and demographic risk factors to identify infants with NAS likely to need pharmacotherapy. METHODS/STUDY POPULATION: A pooled cohort of 761 infants from 5 different studies including 2 trials and 3 observational cohorts will be used to develop the model.

All infants >than or equal to 37 weeks gestational age born to mothers with history of OUD will be included. Infants with congenital disorders and severe medical and surgical illnesses will be excluded. Multivariable mixed effects logistic regression modeling will be performed to predict the need for pharmacologic treatment for NAS. Candidate variables will be included based on clinical knowledge and previously published data. Model performance will be evaluated by measuring discrimination using Area Under the Curve (AUC) statistics and calibration. Model will be internally validated using boot strap validation. RESULTS/ANTICIPATED RESULTS: Pending data analysis DISCUSSION/SIGNIFICANCE OF FINDINGS: Opioid Use Disorder in pregnancy has resulted in concurrent rise in NAS incidence. NAS affects opioid exposed infants variably and accurate prediction of its severity and need for treatment remains elusive. Known clinical and demographic factors can predict the need for NAS therapy in opioid exposed infants, aiding clinical decision making.

OBJECTIVES/GOALS: We will investigate the influence of multisector partnerships in T3-T4 research associated with advances in delivery systems, patient/population outcomes and health policy and the translational processes linked to these improvements. METHODS/STUDY POPULATION: We are using both quantitative and qualitative data to measure and analyze partnership characteristics linked to successful translation into practice & policy. We aim to complete 100 surveys of investigators who have conducted CTSA-supported T3-T4 research to examine partnerships, conditions of collective impact, and quantifiable changes in delivery systems, health outcomes, and policy. Using rigorous criteria, we will select projects for more in-depth interviews to understand the practices of successful translation and roadblocks and barriers that challenge translation. RESULTS/ANTICIPATED RESULTS: The anticipated research products include: (i) an analytic report on partnership structure and processes and the statistical associations to stages of change outcomes, (ii) a series of vignettes to describe the impact stories and translational processes, (iii) cross-project analysis of the data and vignettes to produce generalizable information to improve T3-T4 translation, and (iv) peer-reviewed manuscript(s) for publication. DISCUSSION/SIGNIFICANCE OF IMPACT: The study will inform and improve researcher competencies and accelerate translation in CTSA hubs that emphasize T3-T4 research. We will develop novel definitions of the T3-T4 research impact. Ultimately, the results will inform research training to better address real-world priorities and needs.

Objectives/Goals: Our study’s overarching goal is to characterize the relationship between water transfer rate (Kw) across the blood–brain barrier (BBB) as measured by diffusion prepared (DP) and multi-echo (ME) ASL in two cohorts that have been shown to have regionally different water transfer rates due to underlying changes in BBB physiology. Methods/Study Population: Ten young, healthy participants (aged 21–30 years, 4f) and 12 elderly participants (aged 66–84 years, 8f) underwent MRI scans on a 3T Siemens Prisma scanner. Structural scans, along with DP and ME ASL, were acquired from each of the participants. The order of the DP and ME ASL sequences was reversed in half the participants to account for ordinal bias. FreeSurfer was used to segment the structural image into respective gray matter, white matter, and deep cortical gray regions to perform region of interest (ROI) analysis. Results/Anticipated Results: We are still in the project’s analysis phase. The anticipated result is that we will see different water transfer rate (Kw) patterns between the old and young groups and between the two sequence groups. Discussion/Significance of Impact: The significance of the results is that we can answer two questions: 1) if there are any differences between water transfer rates in the two age groups and 2) whether there are any variations in performance differences between the sequences.

OBJECTIVES/GOALS: Most pregnant youth (ages 14-24) gain more weight during pregnancy than recommended by clinical guidelines. We aim to describe the feasibility and acceptability of home grocery delivery of fruits, vegetables, and healthy snacks to promote healthy weight gain in this vulnerable population. METHODS/STUDY POPULATION: Participants were low-income pregnant youth in Michigan. Each participant was sent biweekly grocery deliveries consisting of $35 worth of fresh fruits, vegetables, and healthy snacks via the app-based delivery service, Shipt. Between deliveries, participants were prompted to respond to weekly text message-based surveys of a 24-hour food recall. This validated nutritional assessment quantifies consumption of fruit and vegetable servings. In addition, participants were asked to send daily photos and descriptions of foods they were eating. This study was approved by the University of Michigan Institutional Review Board. RESULTS/ANTICIPATED RESULTS: To date, 27 participants have been enrolled. Thirteen participants have completed their participation, 4.3 months on average, and were sent an average of 10 grocery deliveries each. In total, over 200 deliveries have been sent with 86% confirmed by the study participant (179/207). Additional outcomes to be assessed include: 1) text message response rates by participants and 2) content from photos and text descriptions of food eaten by participants. The 24-hour recall and text and photo messaging provided in-context data about grocery utilization. DISCUSSION/SIGNIFICANCE OF IMPACT: Grocery delivery is both feasible and acceptable to our youth participants. Use of grocery delivery constitutes a novel intervention to promote healthy weight gain in pregnancy for vulnerable populations through improving access to healthy food options.

OBJECTIVES/SPECIFIC AIMS: Hepatitis C virus (HCV) has a high prevalence among individuals in jail and prisons. Access to HCV treatment has been restricted in jails and prisons. We hypothesized that HCV infection in inmates would be associated with increased mortality in people who were hospitalized while incarcerated. METHODS/STUDY POPULATION: We created and then linked a database of people who were incarcerated and admitted at Lemuel Shattuck Hospital (2004, 2008, 2011) to the Massachusetts Vital Statistic Registry (updated through end of 2015). Death was classified using the Automatic Classification of Medical Entry Death Code. The primary outcome of interest was mortality within 1 year of hospitalization, and the secondary outcome was mortality at any time. The primary indicator of interest was HCV, defined as the presence of the ICD-9 code for HCV on discharge. Covariates included in univariate and multivariate modeling included age, year of admission, and race/ethnicity classified as: White, Black, Hispanic or Other (i.e., Asian, Native American, Multi-Racial, or No answer). RESULTS/ANTICIPATED RESULTS: Of the 1,541 hospital admissions, 21% had HCV, and 57% were white, 22% black, 8% Hispanic and 12% other. Of the 273 total deaths (18% of cohort), 82 deaths occurred within 1 year of hospitalization (5.3% of the entire cohort, 30% of all deaths). The primary cause of death was vascular (21%), followed by chronic liver disease (18%), cancer (17%), overdose/suicide/trauma (19%), pulmonary (7%) and infection (6%). People with HCV were more likely to die of chronic liver disease (40% vs 7%, p<0.001). In the multivariable adjusted model, people with HCV were more likely to die within 1 year of hospitalization (HR 1.59, 95% CI 1.02, 2.49) and more likely to die at any time (HR 1.38, 95% CI 1.06, 1.79). Age, race and gender were not associated with risk of death. Compared to 2004, people admitted in 2008 (HR 2.05, 95% CI, 1.50-2.80) and 2011 (HR 4.02, 95% CI 2.77, 5.83) were more likely to die within 1 year. DISCUSSION/SIGNIFICANCE OF IMPACT: Despite advances in HCV treatment in the community, HCV in inmates is associated with increased mortality.

ABSTRACT IMPACT: The integration of patient-reported outcome measures into clinical care is feasible and can facilitate patient-centered care for individuals with systemic lupus erythematosus. OBJECTIVES/GOALS: Patient-reported outcome measures (PROMs) are powerful tools which can facilitate patient-centered care by highlighting individuals’ experience of illness. The aim of this study was to assess the feasibility and impact of implementing web-based PROMs in the routine clinical care of outpatients with systemic lupus erythematosus (SLE). METHODS/STUDY POPULATION: Outpatients with SLE were enrolled in this longitudinal cohort study at two academic medical centers. Participants completed PROMIS computerized adaptive tests assessing multiple quality of life domains at enrollment and prior to two consecutive routinely scheduled rheumatology visits using the ArthritisPower research registry mobile or web-based application. Score reports were shared with patients and providers before the visits. Patients and rheumatologists completed post-visit surveys evaluating the utility of PROMs in the clinical encounters. Proportions with confidence intervals were calculated to evaluate survey completion rates and responses. RESULTS/ANTICIPATED RESULTS: A total of 105 SLE patients and 16 rheumatologists participated in the study. Subjects completed PROMs in 159 of 184 eligible encounters (86%, 95% CI 81 - 91), including 90% of visit 1’s (95% CI 82 - 95) and 82% of visit 2’s (95% CI 72 - 90. Patients and rheumatologists found that PROMs were useful (91% and 83% of encounters respectively) and improved communication (86% and 72%). Rheumatologists reported that PROMs impacted patient management in 51% of visits, primarily by guiding conversations (84%), but also by influencing medication changes (15%) and prompting referrals (10%). There was no statistically significant difference in visit length before (mean=19.5 min) and after (mean=20.4 min) implementation of PROMs (p=0.52). DISCUSSION/SIGNIFICANCE OF FINDINGS: The remote capture and integration of web-based PROMs into clinical care was feasible in a diverse cohort of SLE outpatients. PROMs were useful to SLE patients and rheumatologists and promoted patient-centered care by facilitating communication.

OBJECTIVES/GOALS: The objective ofthis study is to use GNR technology to track immune cells infiltrating malignant brain tumors that are delivered as part of a novel immunotherapeutic strategy. We seek to implement this new platform to elucidate the underlying mechanisms of therapeutic benefit from ACT via correlation between biodistribution and efficacy. METHODS/STUDY POPULATION: Utilizing the inherent two-photon luminescent signal of GNRs, we will identify uptake and phenotype of lineage negative hematopoietic stem cells (HSCs) in vitro. HSCs will be isolated from the bone marrow of 6-week-old C57bl/6 female mice. Following isolation, HSCs will be co-cultured with varying concentrations of GNRs in DMEM w/o sodium pyruvate for 24 hours, tested for viability, and images to quantify uptake and identify phenotyping. CT contrast of our novel Iodine-capped PEGylated gold nanorods will be confirmed through microCT and biodistribution of HSCs at time points after injection will be identified via CT visualizationin vivo. RESULTS/ANTICIPATED RESULTS: We expect that increased GNR signaling 24 hours post-transplant in the tumors of glioma-bearing mice will be positively correlated with long term survival following ACT. Published data from our labs have revealed that CCR2+ lineage-negative HSCs significantly accumulate in tumor of glioma-bearing mice12. Importantly, CCR2+ lineage-negative HSCs promote differentiation to dendritic cells in the tumor, increase antitumor T cell responses mediated by cross-priming and cross-presentation, and improve efficacy of immune checkpoint inhibition12. Given that HSCs are important in mediating immunotherapy efficacy, we seek to correlate the accumulation of GNR signaling within the tumor as a marker of treatment response. DISCUSSION/SIGNIFICANCE: Adoptively transferred cells have been imaged using numerous published methods. While promising to the field of immunotherapy, these methods lack significant clinical validation. GNRs have not been used to study hematopoietic stem cells in the context of ACT and brain malignancies. Our research is poised to address this gap.

OBJECTIVES/GOALS: We aim to discover safer and more effective therapeutics for CNS disorders. Current therapeutic development is hindered by dosing out drugs for safe consumption. By identifying proteins with narrow functional roles in the brain (i.e., behavioral control), we can develop drugs targeting these proteins for improved treatment safety and efficacy. METHODS/STUDY POPULATION: We focused on an evolutionarily new, non-neuronal, non-synaptic glutamate signaling mechanism, system xc- (Sxc). Sxc activity was eliminated by mutating the gene Slc7a11 through pronuclear injection of zinc-finger nucleases into Sprague Dawley rat embryos to create a line of rats lacking Sxc (MSxc). To confirm Sxc mutation, we verified that tissue from MSxc rats had a complete lack of xCT, which is the regulatory subunit of Sxc that is encoded by Slc7a11. We also verified that astrocyte cultures generated from MSxc tissue lacked cystine-evoked glutamate release. Next, we measured development (body weight), CNS regulation of metabolism, and other indicators of generalized, non-specific brain function as well as behaviors that are reliant on behavioral control, such as impulse control and response inhibition. RESULTS/ANTICIPATED RESULTS: Eliminating Sxc was not lethal and did not impair development or produce widespread changes in brain function as is commonly observed when deleting other glutamate mechanisms. MSxc rats did not differ from wildtype in growth rate, central regulation of metabolism as reflected by absolute or diurnal changes in core body temperature, locomotor activity in a familiar or novel environment, or simple forms of cognition such as novel object recognition, or operant responding (food and cocaine-reinforced). In contrast, behaviors that rely on behavioral control were impaired. MSxc rats displayed deficits in impulse control and behavioral flexibility. We hypothesize that MSxc rats will also show deficits in response inhibition using the stop signal reaction time task, a common metric used in clinical populations. DISCUSSION/SIGNIFICANCE: Eliminating Sxc activity in rats produced deficits in behaviors reliant on impulse control, without impacting development or simple brain function. These results show the potential of targeting Sxc to restore behavioral control without generating therapeutically limiting adverse effects resulting from non-specific changes in brain function.

Objectives/Goals: Baseline lung allograft dysfunction (BLAD) is defined as the failure to attain normal lung function after transplant and has been associated with impaired survival. BLAD has no consensus definition and assessment of varying thresholds of abnormality may identify an impact on survival or development of chronic lung allograft dysfunction (CLAD). Methods/Study Population: This is a retrospective cohort analysis of bilateral lung transplant recipients who were transplanted between 1/1/2012 and 12/31/2022 who have complete pulmonary function data posttransplant. Thresholds of BLAD including percent predicted levels of FEV1 and FVC at 80%, 75%, 70%, 65%, and 60% were assessed. Outcomes evaluated include survival, development of CLAD, and association of key risk factors with the development of BLAD including donor, recipient, operative, and postoperative characteristics. Results/Anticipated Results: Totally, 680 bilateral lung transplant recipients were identified. Prevalence of BLAD ranged from 41.9% to 9.7% at specified thresholds. We anticipate performing survival analyses and evaluating development of CLAD in patients with BLAD at varying thresholds. We are assessing key donor, recipient, operative, and postoperative variables for association with BLAD. Preliminary analyses demonstrate significant associations of BLAD with recipient-donor height mismatch, prolonged mechanical ventilation time posttransplant, increased length of hospitalization posttransplant, the use of cardiopulmonary bypass intraoperatively and surgical allograft downsizing. Discussion/Significance of Impact: A threshold of BLAD at 70% predicted FEV1 and FVC or lower suggests importance for developing CLAD. Key characteristics associated with BLAD suggest importance of height mismatch, operative complexity, frailty, and severity of disease at time of transplant and immediately postoperatively.

OBJECTIVES/GOALS: Our goal was to explore the actions of odorants on mortality and seizures in a DS mouse model (scn1a+/-), which have spontaneous seizures and high rate of SUDEP. We hypothesize that odorants that have actions on olfactory->;extended amygdala pathways will decrease SUDEP, potentially through attenuation of neuronal activation in the extended amygdala. METHODS/STUDY POPULATION: Dravet syndrome mice (heterozygous scn1a+/- ) were exposed for at least eight hours a day to either 2-phenylethanol (2PE, rose odor ), lemon extract, or vehicle odorant in group housed cages. This was repeated daily for 15 days starting at postnatal day 20/21. Mortality in each group was recorded. A subset of 2PE-exposed animals had an extended washout period following odorant exposure to continue to determine the long-term effect of odorant exposure on mortality. RESULTS/ANTICIPATED RESULTS: Our preliminary results show a strong trend for decreased mortality in the 2PE-exposed group (16.1% mortality (n=31) vs 38.5% mortality in vehicle control (n=26), p=0.06, Barnard’s test). Survival analyses show similar results (p=0.056 Kaplan-Meier curve, p=0.046 when removing those animals that died before completing day one of exposure). The lemon scent-exposed animals had a non-significant increase in mortality compared to controls from our preliminary experiments (50% mortality, n=8). Overall, these results suggest that mortality effect is dependent on specific odorants and that this effect is transient. DISCUSSION/SIGNIFICANCE: Our preliminary data support that odorant exposure can decrease mortality in a Dravet Syndrome mouse model, suggesting that more work to determine the mechanism of action and circuitry involved may illuminate new targets and therapies for preventing SUDEP in epilepsy patients.

OBJECTIVES/GOALS: The EWSR1-FLI1 gene fusion is implicated as a source of oncogenic activity in the majority of Ewing sarcoma (EwS) cases (>70%). Our studies will provide unparalleled insight into the transformative mechanics of EWS-FLI1 METHODS/STUDY POPULATION: EWS-FLI1 is an intrinsically disordered protein (IDP). IDPs do not form stable secondary or tertiary folds in biological conditions and ofter contain proline-rich regions and other multivalent binding regions. These regions interact with numerous partners resulting in highly dynamic and complex protein-protein interactions. Recent advances in proximity labelling techniques, such as the use of a biotin ligase fused to a protein of interest are exceptionally well suited to identifying IDP interactomes since they do not rely on the binding affinities but rather the distance between interacting proteins. Combining novel discovery proteomics with Nuclear Magnetic Resonance (NMR) approaches will provide unparalleled insight into the transformative mechanics of EWS-FLI1. RESULTS/ANTICIPATED RESULTS: Using the proximity labeling technique TurboID, which limits the number of false-positive interactions, we have shown that EWSR1 and EWS-FLI1 act within the spliceosome (responsible for mRNA splicing and processing). Using bioinformatics techniques, we show EWS-FLI1 interacts with peptidyl-prolyl isomerase (PPI) proteins, specifically PPIL1, through the EWS-FLI1 N-terminal region (EWS-LCD). We used NMR to provide insight into the interaction between PPIL1 and EWS-LCD, showing that EWS-LCD interacts with the catalytic region of PPIL1. We anticipate that PPIL1 isomerizes EWS-FLI1 to modulate its activity, and that EWS-FLI1 interacts with the spliceosome through formation of a biological condensate. However, future proximity labeling and NMR studies are needed to verify this activity. DISCUSSION/SIGNIFICANCE: Our studies will yield actionable insights regarding the protein-protein interfaces in EWS-FLI1 that can be targeted to attenuate the oncogenic activity of EWS-FLI1. More broadly, our results will be applicable toward understanding the etiology of other pediatric cancers and for guiding the development of novel targeted treatments.