OBJECTIVES/GOALS: Minoritized populations experience a large burden of Alzheimer’s Disease; interventions are often delayed and underrepresented communities’ participation in clinical trial research is low. Lack of information has been proposed as a barrier to clinical research enrollment of minoritized populations. METHODS/STUDY POPULATION: Brain Train, a cluster randomization trial, evaluated the role of education in increasing willingness to participate in research. A 3 segment program was developed whereas segment 3 had two versions: clinical trial education or healthy brain aging video. Brain Train was presented to multicultural communities with participants of at least 50 y/o throughout WNY. The primary outcome measure is evaluating the percent change in responding yes to would you be interested in participating in a clinical trial? before and after segment 3. The secondary outcome measure and exploration of barriers are measured by a Research Attitude Questionnaire. Demographic information such as age, race, gender, socioeconomic status, and educational attainment is collected. The statistical model is a generalized linear mixed model. RESULTS/ANTICIPATED RESULTS: Sixteen sessions with 281 participants were completed to generate the pilot dataset. We see 59.29 % of individuals answered yes before segment 3 and 46.02% answered yes after the educational intervention. Our data shows there is a 13.3% decrease in clinical trial participation interest after the educational intervention. Through our RAQ responses, our data shows significance when it comes to our participants’answers to questions regarding society needing to devote more resources to medical research (p=0.04). Trust emerged as the most significant barrier when it comes to one’s willingness to participate in medical research and clinical trials (p=0.03). DISCUSSION/SIGNIFICANCE: Our preliminary results from the first sixteen events suggest that the power of education is not sufficient to overcome barriers to clinical trial participation for underrepresented communities. Instead, trust appears to be the most significant barrier. Trust building strategies should be explored to answer this research question.

OBJECTIVES/GOALS: Thiswork is an evidential study that demonstrates the positive impactof integrating Community Health Workers (CHWs) and SocialDeterminants of Health on an important health outcome, notably in decreasing the 30-day unplanned hospital ED readmissions at Sinai Health Systems. METHODS/STUDY POPULATION: Using datafrom the Sinai Urban Health Institute (SUHI), we compare predictingthe readmissions of patients with and without data pertainingto Social Determinants of Health (SDoH). We thoroughly describe the data cleaning and data pre-processing, done in collaboration with experts in community health. We use a fundamental and ubiquitous classifier in Random Forest for its feature characterization capability in order to translate models results into insights and recommendations for the CHW program. RESULTS/ANTICIPATED RESULTS: We show that when patients are simply engaged byCHWs, regardless of the content of those conversations, we canincrease the predictive accuracy of our classifier by 5%. We usethis result to make recommendations for improving patient careand discuss limitations and future work. Importantly our workpoints directly to the human connection between patients andCHWs as an important feature in the readmission rate. DISCUSSION/SIGNIFICANCE: Our work shows that the predictive capabilities of the classifier increases with CHW logs and SDoH survey data, highlighting the benefit of collecting this information. This is the first step in early identification of such patients so that CHWs are focusing on and providing resources to patients who will most benefit from the program.

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.

Health disparities between Appalachia and the rest of the country are widening. To address this, the Appalachian Translational Research Network (ATRN) organizes an annual ATRN Health Summit. The most recent Summit was held online September 22–23, 2020, and hosted by Wake Forest Clinical and Translational Science Institute in partnership with the Northwest Area Health Education Center. The Summit, titled “Community-Engaged Research in Translational Science: Innovations to Improve Health in Appalachia,” brought together a diverse group of 141 stakeholders from communities, academic institutions, and the National Center for Advancing Translational Science (NCATS) to highlight current research, identify innovative approaches to translational science and community-engaged research, develop cross-regional research partnerships, and establish and disseminate priorities for future Appalachian-focused research. The Summit included three plenary presentations and 39 presentations within 12 concurrent breakout sessions. Here, we describe the Summit planning process and implementation, highlight some of the research presented, and outline nine emergent themes to guide future Appalachian-focused research.

OBJECTIVES/SPECIFIC AIMS: The aims of this project are to: (1) examine placebo effects on subjective and objective outcome measures, (2) determine if an increase in the placebo is associated with changes in benefit, (3) evaluate if the trauma related insomnia placebo group in our study has different side effect reports compared with insomnia placebo participants in previous suvorexant trials, and (4) (Exploratory) examine associations between the placebo group’s characteristics (e.g., trauma/PTSD severity, demographics) and placebo effects. METHODS/STUDY POPULATION: The parent study is a randomized double-blind placebo-controlled clinical trial (clinicaltrials.gov ID: NCT02704754) of suvorexant for treatment of adults (age 18–55) with insomnia that started or worsened after trauma exposure. Suvorexant is a first in class orexin antagonist and is approved by the FDA for the indication of insomnia. In this 6-week trial, all participants initially take 10 mg of suvorexant/placebo, and the dose will be increased to 20 mg if participants continue to experience clinically significant insomnia symptoms 1 week after starting the medication. Sleep outcomes will be measured by polysomnography, daily sleep diary, and the Insomnia Severity Index. RESULTS/ANTICIPATED RESULTS: We hypothesize that (1) within the placebo group data both subjectively and objectively measured outcomes will similarly show improvement in insomnia symptoms, (2) the increase of the placebo medication dose will result in an increased benefit, (3) the trauma-related insomnia placebo group will have the same type and similar rate of side effects reported in previous suvorexant trials. DISCUSSION/SIGNIFICANCE OF IMPACT: Most previous studies examining placebo effects focused on pain and depression. Information obtained from this project will complement our current understanding of placebo effects by characterizing placebo effects on trauma-related insomnia. This study will inform the development of novel strategies to maximize utility of placebos in future clinical trials.

OBJECTIVES/GOALS: We are launching a multi-center prospective registry for patients with metastatic invasive lobular carcinoma (ILC), the second most common type of breast cancer, to better understand patterns of progression, imaging features of metastatic sites, and if serial cell free DNA measurements can serve as a surrogate marker of disease progression. METHODS/STUDY POPULATION: Patients with biopsy proven metastatic ILC of any receptor subtype will be included in the registry. We will exclude patients with ductal histology only or those with multiple primary malignancies. Patients will be enrolled at four large academic medical centers across the country. Cell free DNA measurements using a tumor informed assay will be obtained every 3 months concurrent with regular clinical imaging. Disease status will be determined by the patient’s medical oncologist by taking into account imaging, tumor markers, symptoms, and cell free DNA measurement. At each time point, patients will be surveyed on their quality of life and their medical oncologists will be asked to rate the clinical utility of the cell free DNA value. Patients will be followed indefinitely. RESULTS/ANTICIPATED RESULTS: We will explore whether the use of serial cell free DNA or a combination of blood-based biomarkers and clinical endpoints can reliably identify treatment response and disease progression in patients with metastatic ILC. Many patients with metastatic ILC have unmeasurable disease on imaging and are thereby excluded from clinical trials. The end goal of this registry is to determine if blood-based biomarkers can be used as a proxy for measurable disease in ILC patients and therefore increase clinical trial enrollment for this subgroup of patients. DISCUSSION/SIGNIFICANCE: The creation of this prospective registry will open the door for future studies of blood-based markers that reflect disease stability and progression, which is an unmet need specifically in ILC. Identification of such markers could lead to a novel treatment response endpoint, changing the way patients are enrolled in trials and managed clinically.

OBJECTIVES/GOALS: Vascularized composite allotransplantation (VCA) restores devastating soft tissue injuries. However, graft viability may be compromised during ischemia time, thus preservation techniques continue to evolve. Here we summarize our preliminary findings from preservation techniques utilizing hyperosmolar extracellular solution (HES) and electric stimulation in a 6 hour ex-vivo perfusion model. METHODS/STUDY POPULATION: A published, MGH rodent hindlimb ex-vivo perfusion model was utilized for this project. Three baseline control elements were taken to compare our results including: a baseline muscle biopsy, harvested hindlimb preserved on ice(4°C) in static cold storage (SCS), and 6 hour perfusion (SNMP) were used to compare results of our four aims. The four aims are shown in Table 1. HES was composed of muscle media and the addition of mannitol until 3 concentrations were made: 300, 500, and 800 mOsm concentrations. In aim 4, the perfusate composition was changed to test a hyper-oncotic purfusate. After 6 hours a muscle biopsy was taken to analyze energy cofactors via liquid chromatography-mass spectrometry, referred to as energy charge. Weight gain (edema), lactate levels, oxygen consumption and energy charge (EC) were used as markers for muscle tissue viability. RESULTS/ANTICIPATED RESULTS: In Aim 1, the higher osmolarity of HES indirectly reduced weight gain but consequentially reduced the EC below 5% when compared to SCS control group. We next incorporate HES into the perfusion model, Aim 2, and noticed a diminution in weight gain. The 500 mOsm group had substantial improvement in EC, lactate production and improved oxygen exchange when compared to a controls: fresh muscle biopsy and SNMP. In Aim 3, after a 6 hour perfusion with the addition of electric stimulation, graft edema improved by 10%, EC improved by 23% and O2 dissociation was highest of all 4 aims. Consequentially, due to muscular contraction the lactate levels were highest. In Aim 4, using a hyper-oncotic perfusate, edema reduced the most during the 6 hour perfusion but revealed lower EC and similar lactate/O2 results. However when left on for 24 hours, edema was significantly higher with lactate build up, EC improved with time as well as O2 dissociation. DISCUSSION/SIGNIFICANCE OF IMPACT: Here we have shown our preliminary results comparing our known ex-vivo perfusion model to multiple hypothesis to improve VCA graft viability. These preservation techniques demonstrate promising results but further studies are ongoing to confirm this encouraging outcome.

OBJECTIVES/GOALS: To determine the relationship between race/ethnicity, geospatial (place-based) social determinants of health (SDOH; rurality and child opportunity index (COI)), and asthma-related adverse events (AAE: hospitalizations, emergency department (ED) visits) among children with asthma in Arkansas. METHODS/STUDY POPULATION: Using the Arkansas All-Payer Claims Database, we conducted a retrospective analysis of children (5-18 years). Medicaid-enrolled children with, 1 asthma diagnosis (ICD-10 J45.xx) for any type of medical event in 2019 were included. Race/ethnicity were self-reported (non-Hispanic White, non-Hispanic Black, Hispanic/Latino). Due to small sample size, all other racial/ethnic groups were classified as Other. Rural-Urban Commuting Area (RUCA) codes were used to determine rural-urban designation using 4-category classification by zip code. COI level was determined by zip code (scale: very low- to very-high opportunity). AAEs were identified using 2019 medical claims. RESULTS/ANTICIPATED RESULTS: The cohort (n=25,198) included 38.7% White, 32.9% Black, 6.0% Hispanic, 5.1% Other, and 17.3% Missing race/ethnicity children. Overall, 61.2% live in rural and 38.8% live in urban areas. Among rural children, 33.1% were in very-low, 34.4% low, 20.8% moderate, 11.6% high, and 0.1% very-high opportunity areas. Among urban children, 32.6% were in very-low, 12.4% low, 17.5% moderate, 19.5% high, and 18.0% very-high opportunity areas. Overall, Black children more frequently lived in very-low or low opportunity areas (75.4%). Among rural children, 9.3% had an AAE. White children had highest rates of AAE. Overall, AAE rates were variable by rurality/urbanity and COI level. DISCUSSION/SIGNIFICANCE: Differences in asthma outcomes by race/ethnicity, rurality, and COI level were unexpected, with similar rates of poor outcomes across the cohort. These finding underscore the complexity of the relationships between race/ethnicity, geospatial SDOH, and asthma outcomes.

OBJECTIVES/GOALS:

• A web interface that allows for easy upload of CSV text data to time-based visualizations

• Implementation of change points analysis to identify and display points where event rates increased or decreased

• customizable plots where the user can change point shapes, color, etc.

• customizable and advanced filtering support

• support for plot comparisons and exports

METHODS/STUDY POPULATION: We used the R/Shiny framework to develop a web application for visualization of time stamped data. The Research and Exploratory Analysis Driven Time-data Visualization (READ-TV) application allows for user-friendly mining for longitudinal patterns in data. READ-TV is built specifically for FD analysis, but is easily adaptable to other clinical use cases, as we allow for the use of general metadata on events and cases.The building of a quantitative framework for event analysis starts with the application of homogeneous Poisson processes, which relate the times of occurrence of events in terms of an underlying rate. To understand the changes in this underlying rate, changepoint analysis is used to model the rate as a function of time using piecewise constant approximations. The changepoint analysis allows us to identify the specific periods of time where the rate of FD is increased relative to a baseline or a desired operating range. RESULTS/ANTICIPATED RESULTS: READ-TV application allows for import of time stamped event data from multiple cases. Event and case metadata are supported to facilitate filtering and mining of interesting subsets of data. Stem plots are used for visualization of selected event timelines in chosen cases. This visualization is accompanied with summary of the number and estimates of rates of occurrence of specific event types (e.g. types of FD). Change-point analysis is implemented using the ‘changepoint‘ R library. These analyses allow the users to quickly understand whether the rates of events (FD) is changing across the case timeline and where exactly these changes are occurring. DISCUSSION/SIGNIFICANCE OF IMPACT: We have demonstrated the READ-TV application to the team of the AHRQ-funded Human Factors and Systems Integration in High Technology Surgery (HF-SIgHTS) study. The ability to visualize and perform quantitative analysis of the study data was received with unanimous positive feedback and enthusiasm. We continue READ-TV development focusing on (1) increased user-friendliness using the HF-SIgHTS as our focus group, (2) increased functionality, and (3) use of more general localization terminology to allow for other applications.

OBJECTIVES/GOALS: Multivariate regression is used for surgical outcomes analyses; but does not allow for evaluation of all variables. Machine learning could be the perfect alternative to address this issue. Our aim was to evaluate whether machine learning is a feasible alternative to evaluate surgical outcomes. METHODS/STUDY POPULATION: H-CUP National Inpatient Sample database was queried for adult patients with colorectal cancer who underwent colorectal resection, while the NSQIP database was queried for adult patients with rectal cancer who underwent proctectomy. A multivariate regression analysis was performed to assess risk factors associated with 30-day complications following those procedures. Subsequently, machine learning techniques of under-sampling and oversampling were applied to the same datasets for the evaluation of risk factors for the same outcome. These techniques were used to achieve a larger population sample size and to detect statistical significance. Results between the two methodologies were compared. RESULTS/ANTICIPATED RESULTS: Multivariate regression revealed that open approach, gender, race, geographic location, number of comorbidities, and type of insurance was associated with increased 30-day mortality in colorectal resection patients. Conversely, the use of machine learning revealed that preoperative weight loss, preexistent chest heart failure, renal failure or perivascular disease were strongly associated with 30-day mortality. For proctectomy patients, multivariate regression found no association between surgical approach and 30-day mortality. However, machine learning revealed gender, hypertension, and reoperation to be strongly associated with 30-day mortality. DISCUSSION/SIGNIFICANCE OF IMPACT: Machine learning enabled multiple combinations that were not possible to examine in a conventional multivariate regression analysis. Machine learning compared to traditional multivariate regression produced significantly different outcomes, highlighting the need for in depth of these methodologies.

OBJECTIVES/SPECIFIC AIMS: The purpose of the current study is to determine predictors of linkage-to-care (LTC) outcomes among patients screened for hepatitis C virus (HCV) infection in the emergency department (ED). The study is one of the first to report the impact of factors influencing medical eligibility for HCV treatment. By establishing predictors of linkage outcomes, we hope to characterize patients at highest risk of being lost to follow-up. This information can be used to guide the development of targeted interventions and optimize linkage efforts at institutions with similar programs. METHODS/STUDY POPULATION: In 2013, we implemented an opt-out HCV screening program targeting baby boomers presenting to the ED. The screening program was expanded in 2015, and all patients who presented to the ED (an academic ED with 73,000 annual visits) between 2015 and 2018 were offered an HCV antibody test. Confirmatory RNA tests were ordered for all patients with a positive screening test, and those with confirmed infection were contacted (when possible) by a linkage coordinator, who sought to facilitate linkage to outpatient care. Patients were followed until they had been successfully linked to care or were lost to follow-up; successful linkage was defined as a patient attending a scheduled HCV clinic visit. Linkage outcomes (linked vs. not linked) among confirmed cases were analyzed based on previously described LTC barriers, sociodemographic factors, and factors influencing HCV treatment eligibility. Binary multiple logistic regression was used to model the relationship between predictors and LTC outcomes. RESULTS/ANTICIPATED RESULTS: Between August 2015 and June 2018, 48,947 patients were screened for HCV, yielding 4,026 positive results. HCV infection was confirmed in 1,798 patients, 1,651 of whom were eligible for the current study. By July 2018, 884 eligible patients (53.5%) had been successfully linked to care. Odds of linkage failure were decreased among patients with known linkage barriers including mental health disorders (adjusted odds ratio [aOR], 0.28; 95% confidence interval [CI], 0.21-0.37), active substance abuse (aOR, 0.65; 95% CI, 0.47-0.91), and ‘competing’ medical comorbidities (aOR, 0.31; 95% CI, 0.21-0.47); however, odds of linkage failure were increased in the presence of medical comorbidities leading to ineligibility for HCV treatment (aOR, 21.71; 95% CI, 7.02-67.13). Decreased odds of linkage failure were also found among patients belonging to the baby boomer population (aOR, 0.64; 95% CI, 0.46-0.91) and individuals with HBV co-infection (aOR, 0.36; 95% CI, 0.14-0.94). Finally, odds of linkage failure were increased among patients without health insurance (aOR, 1.952; 95% CI, 1.01-3.79) and with public health insurance (aOR, 2.140; 95% CI, 1.08-4.24). While African-American patients exhibited decreased odds of linkage failure in bivariate analyses, this association was not significant in the multivariate analysis. Variables ultimately found not to be predictive of linkage outcome included race, ethnicity, gender, homelessness, and incarceration. DISCUSSION/SIGNIFICANCE OF IMPACT: In contrast with current trends in the literature, our analysis found that patients with any of three well-described linkage barriers (mental health disorder, active substance abuse, and ‘competing’ comorbidities) exhibited decreased odds of linkage failure. It is possible that knowledge of these barriers by linkage personnel led to increased persistence of efforts for such patients, resulting in improved linkage among patients with these barriers. However, another possibility is that individuals with factors such as a mental health disorder or medical comorbidity were more likely to experience a clinical encounter, during which discussions about HCV treatment could take place. It is also possible that other contributing factors were unaccounted for by our analysis, such as internal programmatic issues, patient education and income levels, marital status, or other ‘destabilizing’ factors that would reduce a patient’s likelihood of being contacted by a linkage coordinator. While patients with general medical comorbidities were less likely to experience linkage failure, patients who were ineligible for HCV treatment due to a medical comorbidity were significantly more likely to experience linkage failure. While this finding is intuitive, it is nonetheless troubling, as we would expect patients with severe medical comorbidities to benefit greatly from health counseling, mental health services, substance abuse treatment, or other resources available through linkage. As a result of this finding, we intend to adapt our approach to medically ineligible patients, emphasizing the availability and potential usefulness of such resources. It is unsurprising that uninsured and publicly insured patients were less likely to be successfully linked to care, as HCV treatment continues to be associated with significant financial costs for such patients. However, it is also possible that confounding factors such as education and income level, which were not included in our analysis, would be more predictive of patients’ ability or willingness to undergo HCV treatment. More research is needed to improve linkage-to-care for patients with HCV. In the near future, we intend to conduct an internal evaluation, as well as a positive deviance analysis of other LTC programs, to determine additional ways to improve linkage outcomes for all patients screened for HCV in the ED.

OBJECTIVES/SPECIFIC AIMS: To establish an in vitro quantitative method for the evaluation of polymeric film disintegration that can be applied to predict in vivo behavior. METHODS/STUDY POPULATION: Two clinically advanced vaginal microbicide film products containing tenofovir and dapivirine were used as model films throughout this work. Films were made using the solvent cast manufacturing method in which polymers, excipients, plasticizer, and APIs were either dissolved or dispersed in water, mixed, and cast on a heated substrate. The novel, quantitative method was developed using a TA.XT Plus Texture Analyzer® (Texture Technologies) in combination with a TA-108S5 fixture and the TA-8A: 1/8″ diameter rounded end ball probe. Exponent® was used as the data analysis software. In this method, the film was placed and secured in the fixture, the probe applied a constant force to the film product, and a biologically relevant amount of fluid was applied to the film. The probe was able to penetrate the film upon disintegration resulting in an applied force of zero at that point. A curve of force Versus time was plotted, and disintegration time was defined as the time between fluid addition until the probe force reached zero. Test parameters were optimized in order to reduce error. Visual observation of film disintegration was conducted in the in vivo macaque model using films that included a water-soluble blue dye for film visualization. Colpophotography was also used to confirm film disintegration. In vitro results were compared with in vivo findings. RESULTS/ANTICIPATED RESULTS: The Texture Analyzer disintegration method developed provided quantitative disintegration times and did not rely on user defined endpoints which is common in many visual disintegration tests. The disintegration method was able to distinguish differences between the 2 clinical film products and produced reproducible disintegration times for the tenofovir and dapivirine films. The tenofovir film had a shorter disintegration time (41.28±2.85 s) compared with that of the dapivirine film (88.3 6±9.82 s). This method was also able to distinguish changes made to these 2 clinical film products in terms of volume and formulation alterations. In vitro and in vivo disintegration times differed by orders of magnitude, with in vitro time being measured in seconds and in vivo time being measured in days, for a variety of factors, mainly the application of constant force to the film product. Regardless of these differences, the rank order of film disintegration remained constant for in vitro and in vivo disintegration and an In Vitro In Vivo Correlation (IVIVC) trend could be seen. DISCUSSION/SIGNIFICANCE OF IMPACT: Standardization of preclinical in vitro assessments which minimize user bias are crucial to the field of pharmaceutical film development. As this field continues to develop and more products advance for pharmaceutical application, this method has the potential to become a standard assessment of film functionality. This study represents a first step in the process of developing an IVIVC. More films will need to be tested using both in vitro and visual methods in order to establish and accurate factor to predict in vivo behavior.

A number of publications have discussed approaches to training the scientific workforce in comparative effectiveness research (CER) and patient-centered outcomes research (PCOR). To meet this need, funders have offered resources for developing educational materials and establishing training programs. To extend these efforts into specific researcher communities, the Agency for Healthcare Research and Quality developed an R25 Funding Opportunity Announcement that called for basic, advanced, and experiential training for a specific researcher community in collaboration with associated program partners. This paper describes the strategies developed by the 5 subsequently funded programs, their specific researcher communities and program partners, and the challenges associated with developing in-person and online programs. We focus on lessons learned that can be translated into developing training programs nationwide and on training for the special populations of interest. We also discuss the creation of a sustainable network for training and the conduct of comparative effectiveness research/patient-centered outcomes research in targeted communities.

OBJECTIVES/SPECIFIC AIMS: This study aims to understand the potential immunomudualtory effect of APOL1 variants in auto-antigen activated myeloid cells by assessing lysosomal integrity in activated cells expressing APOL1. The primary stimuli were: 1. ssRNA hY3 as a proxy for the Ro immune complex; 2. in an bulk RNA seq model, interferon-response gene, Siglec 1, as a read out of interferon activity. The primary outcomes were: 1. Myeloid cell APOL1 expression both in primary macrophage cultures and ex-vivo patient derived macrophages; 2. Lysosome integrity as measured by fluorescence intensity of lysotracker dye on light microscopy. METHODS/STUDY POPULATION: All recruited subjects provided written informed consent as per the NEW YORK UNIVERSITY Division of Rheumatology-wide Specimen and Matched Phenotype Linked Examination (SAMPLE) protocol. Subjects were African American; SLE subjects met 4 American College of Rheumatology criteria for SLE. Healthy donor monocytes representing each genotype in duplicate (reference allele: G0/G0; heterozygote variant: RV/G0; and homozygote variant RV/RV) were cultured with GM-CSF to yield macrophages which were incubated in serum free media or with hY3 ssRNA (TLR 7/8 agonist) to yield inflammatory M1 macrophages. Fold increase of APOL1 in untreated vs hY3 treated macrophages was measured using qPCR. Live cells were then cultured on glass chamber slides with DNA dye, DAPI, and Lysotracker red, a fluorescent dye that stains acidic lysosomes. As a proof of concept, interferon response gene, Signlec1, and APOL1 transcriptional activity in peripheral blood monocytes (PBMCs) were measured and correlated in 17 SLE patients by RNA seq. RESULTS/ANTICIPATED RESULTS: Regardless of genotype, hY3 increased APOL1 expression by 29 (+/−18.4) fold (P = 0.007 vs no treatment). Genotyping of the qPCR product showed concordance with the chromosomal DNA with the RV heterozygotes expressing both alleles. To examine lysosomal membrane integrity, live hY3-treated macrophages were stained with lysosotracker dye and fluorescence intensity was measured. Compared to reference allele carrying macrophages, each additional variant allele corresponded with a lesser degree of lysosome compartment staining. In SLE PBMCs, we found that APOL1 was highly expressed, and significantly correlated with Siglec1 (F=10.5; P = 0.005) supporting an association between circulating interferons and APOL1 accumulation in monocytes. DISCUSSION/SIGNIFICANCE OF IMPACT: Given that the “cytokine milieu” in SLE elicits APOL1 expression, induces inflammatory cell metabolic rewiring, and stimulates autophagy thereby exposing defects in autophagic flux, this gene-environment interaction may underpin the relationship between chronic inflammation and heightened APOL1 polymorphism-attributed cardiovascular risk. These data support further inquiry into the intersection between chronic autoimmunity and APOL1’s functional role in the vascular microenvironment. The in vitro studies herein extend our prior work by demonstrating a mechanistic link between SLE-associated inflammation, APOL1 risk variant status and CVD via a lysosomal defect which converges on common autophagic and metabolic pathways in mononuclear cells.

OBJECTIVES/SPECIFIC AIMS: N/A. METHODS/STUDY POPULATION: N/A. RESULTS/ANTICIPATED RESULTS: N/A. DISCUSSION/SIGNIFICANCE OF IMPACT: There is an increasing need to foster cross-disciplinary research to address complex problems within healthcare. The Sinai Team-based Translational Education Program: the URM Propeller (STTEP-UP) is a NCATS funded program through the Icahn School of Medicine at Mount Sinai. Its goal is to facilitate URM post-doctoral trainees becoming innovative leaders in clinical and translational research. The program includes a team-based research component, where fellows collaborate on a project. This year, disciplines represented by the four fellows include Cardiology, Psychiatry, Neurology, and Pediatrics. Identifying a clinical question and designing an investigation was facilitated by group brainstorming meetings with program mentors. Fellows designed a project to identify medical testing and prescribing that were not clinically indicated throughout the healthcare system, with the goal of exploring whether an intervention, including provider education, could reduce ordering practices. In addition to regular in-person meetings, a licensed virtual learning environment and free web-based sharing platform were used to foster collaboration. Challenges faced throughout this process, included fellows struggling to find protected time, difficulties accessing broad sets of data across the healthcare system, and overcoming administrative barriers between departments. Strengths of this approach, included fellows learning new research strategies and feeling a deeper sense of commonality with their peers. Overall, this experience supports the idea that cross-disciplinary research improves the collaboration and education of emerging researchers. However, addressing logistical and systems-based barriers may better facilitate this education and research.

OBJECTIVES/GOALS: Klotho is a protein linked to improved cognition in aging adults. A specific KLOTHO gene variant, KL-VS, increases circulating levels of Klotho. The current study aims to identify if the KL-VS haplotype and Klotho levels are associated with improved neurocognition in pediatric brain tumor survivors. METHODS/STUDY POPULATION: We are actively accruing pediatric brain tumor patients at UCSF alongside an existing multi-institutional cohort study investigating radiation-induced vasculopathies and cognitive outcomes in this population. Normal controls are being enrolled in parallel. Each patient undergoes: 1) single nucleotide polymorphism genotyping to identify KL-VS haplotype status, 2) enzyme-linked immunosorbent assays to measure circulating Klotho, 3) neurocognitive assessments with a computer-based, validated Cogstate battery, and 4) brain volume and white matter lesion segmentation analyses using MRI sequences obtained as part of routine care. RESULTS/ANTICIPATED RESULTS: Genotyping has been performed on 99 enrolled patients. KL-VS heterozygosity was seen in 22.7% of patients. To date, KL-VS status is not associated with neurocognitive outcomes at baseline or Year 1 testing. Association between KL-VS status, circulating Klotho levels, neurocognitive outcomes, brain volume and white matter lesion segmentation analyses is ongoing. We hypothesize that elevated Klotho levels will be associated with improved neurocognition, increased brain volumes in regions of interest and decreased white matter lesion volumes. DISCUSSION/SIGNIFICANCE OF IMPACT: If circulating Klotho leads to improved neurocognition in pediatric brain tumor survivors, Klotho levels might serve as a prognostic biomarker. Furthermore, as Klotho is being investigated for therapeutic indications, it may represent an intervention to prevent cognitive deficits in these patients.

OBJECTIVES/GOALS: Mobile health applications are widely used by the public but vary in how they are classified and regulated. This study examines the evidence of the safety and efficacy of mobile medical applications specifically focusing on those that are used to manage diabetes. METHODS/STUDY POPULATION: To understand the current regulatory landscape of mobile health applications (mHealth apps) for diabetes, a literature survey was conducted using the Pubmed database. Top mHealth apps were identified by searching the Apple store website using 10 key terms associated with diabetes management applications. A maximum of ten results, when available for each key term, were studied by exploring the FDA databases to understand how the products were regulated and if any were subject to recalls. These selected mHealth apps were also searched on clinicaltrials.gov to see if there were ongoing or completed clinical trials and if the trials were designed to include efficacy and safety outcome measures. RESULTS/ANTICIPATED RESULTS: Of the 71 mHealth apps for diabetes management that were identified, 16 were regulated. These products spanned a diverse range of functions including device data and decision support systems. Although 11 had clinical trial data demonstrating efficacy, only 4 had data demonstrating both efficacy and safety. Two of the regulated applications were subject to product recalls due to programming errors that resulted in incorrect insulin dose recommendations. These two applications had clinical trials evaluating efficacy but not safety. The companies noted that the incorrect insulin calculation from their respective mHealth app could cause either a low- or high-impact hypoglycemic event. DISCUSSION/SIGNIFICANCE OF IMPACT: With little to no clinical trial data to support their safety and efficacy, mHealth apps in the diabetes marketplace pose risks for patients as evidenced by recent safety-related recalls. The results of this study indicate that these products may need to be more tightly regulated.