OBJECTIVES/SPECIFIC AIMS: Immune cells within the tumor microenvironment (TME) play an important role in the development and progression of non-small cell lung cancer (NSCLC). However, data evaluating the impact of individual immune cell types on NSCLC outcomes is limited and often conflicting. We performed a meta-analysis of existing data and used The Cancer Genome Atlas (TCGA) to evaluate the effect of several immune cells on surgical outcomes of stage I-IIIA NSCLC. METHODS/STUDY POPULATION: PubMed was searched to identify eligible studies evaluating survival of surgically resected stage I-IIIA NSCLC patients according to immune cell infiltration. Meta-analysis was performed using a linear mixed-effects model to determine overall, disease specific and progression free survival. We then used a similar patient subset found in the TCGA to validate the meta-analysis findings. For the TCGA analysis, sample-specific scores for different immune cells were computed via xCell using level three RNAseq data. After stratifying the cohort by histologic subtype, the association between each cell type and survival was assessed via Cox Regression, while adjusting for stage, gender and smoking status. RESULTS/ANTICIPATED RESULTS: From the meta-analysis (37 articles eligible; N = 8,162 patients), high levels of CD20+ B cells (hazard ratio [HR]: 0.36, 95% confidence interval [CI]: 0.15-0.85), natural killer (NK) cells (HR: 0.64, 95% CI: 0.41-1.0), and dendritic cells (0.34, 95% CI: 0.13-0.84) were significantly associated with better overall survival (OS); T regulatory cells (HR: 1.85, 95% CI: 1.35-2.54) were associated with worst OS. High CD8+ T cell infiltrates were associated with improved disease-free survival (DFS; HR: 0.85, 95% CI 0.73-0.99), while CD68+ macrophages (HR> 2.83, 95% CI: 1.28-6.24) were associated with worst DFS. In the TCGA cohort, lung adenocarcinomas rich in CD4 T cells, CD8 T cells, B cells, and NK cells were associated with improved OS in unadjusted analysis. In adjusted analysis, only NK cells were associated with improved OS (HR: 0.82, 95% CI: 0.69-0.98). There was no significant association of any immune cell type for DFS in lung adenocarcinomas and with both OS and DFS in Squamous Cell Lung Cancers (p>0.05 for all comparisons). DISCUSSION/SIGNIFICANCE OF IMPACT: The presence of tumor infiltration by specific immune cell subsets may potentially predict survival outcomes in resected stage I-III NSCLC patients. However, the impact of immune cells may not be similar in all histologic types and after adjusting for important clinical confounders.

OBJECTIVES/SPECIFIC AIMS: Aim 1: To evaluate whether psoas muscle size on CT imaging can be used as univariate predictor for increased risk of morbidity and mortality in trauma patients 65 years or older with rib fractures. Primary outcomes will be 30 day mortality. Secondary outcomes will include length of stay, 30 day readmission rate, need for operative/procedural intervention, ICU days, ventilator days, discharge to rehabilitation. Aim 2: An eventual goal of the project will be to use the results of the single variable psoas size study to inform the development of a predictive model for readmission rate in this population based on clinical variables present at admission. METHODS/STUDY POPULATION: This retrospective cohort study will utilize the Maine Trauma Registry to conduct a database review for all persons 65 years of age and older admitted to Maine Medical Center between January 1, 2015 and December 31, 2017 with rib fracture as diagnosed by CT imaging. Psoas caliber will be measured on admission CT. Patient outcomes will be assessed via EMR review. RESULTS/ANTICIPATED RESULTS: Anticipate finding a relationship between decreased psoas caliber and increase in 30 day mortality and post trauma complications. DISCUSSION/SIGNIFICANCE OF IMPACT: If a relationship is demonstrated between decreased psoas caliber and poor outcomes in elderly patients with rib fractures, this early indicator could be used to identify those patients at most risk, for whom targeted interventions may make the greatest difference. Knowing a measure of frailty could also help guide goals of care discussions, because it would allow clinicians to have a more detailed understanding of a patient’s baseline. Those patients identified as frail could be admitted to an ICU level of care and more closely monitored and treated. Alternatively, some frail patients and their families may choose to focus more on comfort and quality of life after achieving a better understanding of a patient’s frailty and risk, changing the direction of care provided. Being able to identify the higher risk patients with an objective measure would allow clinicians to provide more personalized medicine.

OBJECTIVES/SPECIFIC AIMS: Projected to soon become the most prevalent cause of End-Stage Liver Disease, the frequency of Non-Alcoholic Fatty Liver Disease (NAFLD) has been rising in the US community. However, studies on NAFLD among inpatients are lacking. Aims: To report the 1) prevalence and trends, 2) outcomes of NAFLD associated hospitalizations in the US. METHODS/STUDY POPULATION: NAFLD cases were identified in the National Inpatient Sample (2007-2014) with ICD-9-CM codes, and the prevalence and trends over the 8-year period were calculated among different demographic groups. After excluding secondary causes of hepatic fat accumulation from the NAFLD cohorts (n = 210,660), the impact of sex, race, and region on outcomes (mortality, discharge disposition, length of stay [LOS] and cost) of NAFLD was computed with generalized estimating equations (SAS 9.4). RESULTS/ANTICIPATED RESULTS: Admissions with NAFLD tripled from 2007-2014 at an average rate of 79/100,000 hospitalizations/year (p-value < 0.0001), with a larger rate of increase among males vs. females (83/100,000 vs. 75/100,000), Hispanics vs. Whites vs. Blacks (107/100,000 vs. 80/100,000 vs. 48/100,000), and governmental-/un-insured patients vs. privately-insured (94/100,000 vs. 74/100,000). Males had higher mortality, LOS and cost than females. Blacks had longer LOS and poorer discharge destination than Whites; while Hispanics and Asians incurred higher cost than Whites. Uninsured patients had higher mortality, longer LOS and poorer discharge disposition than the privately-insured. DISCUSSION/SIGNIFICANCE OF IMPACT: Hospitalizations with NAFLD is rapidly increasing in the US, with a disproportionately higher burden among certain demographic groups. Measures are required to arrest this ominous trend and to eliminate the disparities in outcome among patients hospitalized with NAFLD.

OBJECTIVES/SPECIFIC AIMS: The objective of this study was to reconstruct patient-specific distal airway patterns at the tissue- and single-cell resolution and develop personalized distal airway models based on utilization of patient-derived DABCs and autologous region-specific stromal cells. METHODS/STUDY POPULATION: Patient-specific distal airway units, containing parental small bronchiole (<2 mm in diameter, >12th generation) and daughter airway branches, including pre-terminal/terminal bronchioles, leading to alveoli (3-7 units/lung), were dissected. Epithelial and stromal cells were isolated from these units and processed for ddSeq single-cell RNA-sequencing (n=6 samples). Autologous DABCs and stromal cells were isolated, propagated, biobanked, and used for establishment of patient-specific distal airway models (3D-organoids and air-liquid interface-based airway wall model; n=10 samples). Region-specific tissue patterns were evaluated using immunofluorescence and laser-capture microdissection (LCM; n=6 samples). RESULTS/ANTICIPATED RESULTS: Single-cell-based human distal airway transcriptome map (constructed based on the analysis of >6,500 distal airway cells obtained from 6 subjects) identified physiological and COPD-relevant distal airway differentiation patterns, including distal airway-specific secretory phenotype (DASP) characterized with high expression of secretoglobins 3A2 and 3A1, surfactant proteins SFTPB and SFTPA2, and mucin 1, unique signatures of DABCs, and stromal (fibroblasts, smooth muscle, endothelial cell subpopulations) and immune (macrophage, T cells, B cell, mast cells). Immunofluorescence analysis and LCM confirmed distribution of cell type-specific markers with differential expression patterns of DABC and DASP signatures. Patient-derived DABC-stromal co-culture models reproduced 3 regenerative patterns: 1) physiological (high DABC-clonogenic potency, establishment of polarized differentiated organoids and DASP-expressing epithelia); 2) hypo-regenerative (failure of DABCs to form clones, spheres and mechanically stable differentiated epithelial barrier); and 3) hyperplastic (generation of DABC hyperplasia accompanied in some COPD samples by mucous-cell hyperplasia mimicking in vivo remodeling patterns). DISCUSSION/SIGNIFICANCE OF IMPACT: Patient-specific maps and models of distal airway regeneration patterns have been established in this study, which can be used to identify candidate pathways that mediate disease-relevant airway remodeling and potentially utilized as pre-clinical platforms for developing personalized therapeutic approaches to suppress the progression of distal airway remodeling in chronic lung diseases, including COPD.

OBJECTIVES/SPECIFIC AIMS: Dopamine transporter (DAT1) gene variation is associated with reward-related phenotypes including alcohol response. There is also evidence for a potential moderating role for mu-opioid receptor (OPRM1) gene variation on the relationship between DAT1 variation and alcohol response measures. We aimed at studying the interaction between the DAT1 VNTR and OPRM1 A118G polymorphisms on alcohol consumption and subjective responses among non-dependent drinkers. METHODS/STUDY POPULATION: We employed a progressive ratio (PR) paradigm of intravenous alcohol self-administration (IV-ASA) using the Computer-Assisted Infusion System (CAIS) to assess the motivation for alcohol seeking and consumption in a sample of nondependent drinkers. We used the Drug Effects Questionnaire (DEQ) and Biphasic Alcohol Effects Questionnaire (BAES) to assess subjective response. IV-ASA measures included average breath alcohol concentration (BrAC) and total ethanol infused. Peripheral blood samples were collected for genotyping. Ethics approval was obtained from the NIH Addictions Institutional Review Board. RESULTS/ANTICIPATED RESULTS: Fifty participants completed the PR IV-ASA session after informed consent. There were significant interactions between the DAT1 and OPRM1 genotypes in subjective effects of alcohol. Simple main effects analysis showed that DAT1 10a allele carriers that were also OPRM1 G allele carriers had significantly higher scores for several measures: “feel the drug effects” (F (1,46)=6.573, P = 0.014), “feel intoxicated”(F(1,46)=8.613, P = 0.005) and “feeling high” (F(1,46)=10.889, P = 0.002) in DEQ and higher sedation (F(1,46)=4.575, P = 0.038) in BAES. The genotypes statistically significantly predicted average breath alcohol (F (1,61) =3.295, p=0.044) and total ethanol infused(F(1,61)=3.632, p=0.032. DAT1 VNTR and OPRM1 A118G polymorphisms taken together accounted for 6.9 and 7.8% of variations in average breath alcohol and total ethanol infused respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: Polymorphic variations in DAT1 and OPRM1 interact with each other in determining subjective effects of alcohol in intravenous alcohol infusion assessing motivation for alcohol seeking and consumption in nondependent drinkers. These epistatic interactions in subjective effects of alcohol are salient in the context of predicting and understanding neurobiological effects of alcohol and thereby the therapeutic responses in treating alcohol use disorders.

OBJECTIVES/SPECIFIC AIMS: This study attempts to evaluate the drinking patterns and traits of individuals who partake in high intensity drinking, defined as binge drinking at 2 or more times the minimum binge count (4 drinks for females, 5 drinks for males). METHODS/STUDY POPULATION: We analyzed data from non-treatment seeking volunteers enrolled in NIAAA screening protocols. The sample included 706 males and 474 females ranging in age from 18 to 91. Subjects were assigned to one of four groups (Non-Binge, Level 1, Level 2, Level 3) based on the highest binge session reported in their Timeline Followback questionnaire. The criteria for each group were different for males and females based on the current NIAAA definitions of binge drinking. The cutoffs for females were 0-3 drinks for Non-Binge, 4-7 drinks for Level 1, 8-11 drinks for Level 2, and 12+ drinks for Level 3. The male drink cutoffs were 0-4, 5-9, 10-14, and 15+ respectively. We looked at various drinking measures (Timeline Followback, Self-Reported Effects of Alcohol (SRE), Alcohol Use Disorders Identification Test (AUDIT)) and trait measures (UPPS-P Impulsivity Scale, Barratt’s Impulsiveness Scale, Buss Perry Aggression Questionnaire) to identify mean differences between groups. RESULTS/ANTICIPATED RESULTS: There were significant differences in drinking patterns between the groups for both males and females. Number of drinking days, average drinks per drinking day, and number of heavy drinking days all increased as binge level increased. There were also significant differences between groups in males for trait measures. Level 2 and Level 3 bingers scored significantly higher on impulsivity and aggression than the Level 1 and Non-Binge groups. Ongoing analyses are examining differences among binge groups on other measures including SRE and AUDIT. Future analyses will explore potential mechanisms underlying the relationships between trait measures and binge drinking using structural equation modeling. DISCUSSION/SIGNIFICANCE OF IMPACT: This study found significant differences between high-intensity drinkers, or “super bingers”, and lighter binge and non-binge drinkers. Super bingers showed an overall heavier drinking pattern across measures. The elevated aggression, impulsivity, and overall heavy drinking patterns of super bingers suggest a behavioral profile that makes this group in particular at higher risk for developing alcohol use disorder and related problems. These traits and behaviors may also help identify targets for treatment interventions for alcohol use disorder.

OBJECTIVES/SPECIFIC AIMS: Glioblastoma (GBM) is a brain cancer with a devastatingly short overall survival of under two years. The poor prognosis of GBM is largely due to cell invasion and maintenance of cancer initiating cells that evade the brain’s innate and adaptive immune responses which enables escape from surgical resection and drives inevitable recurrence. While targeting the brain’s immune microenvironment has long been proposed as a strategy for treating GBM, translational progress has been slow, underscoring the need to investigate the brain’s immune microenvironment for therapeutic avenues. METHODS/STUDY POPULATION: Recent advancements in tunable synthetic immunomodulatory gene circuits targeting metastatic cancers has demonstrated the novel ability to use engineering principles to induce infiltrative cancer cells to express combinatorial immunomodulatory outputs that enable T-cell killing4. Our central hypothesis is: we will be able to significantly improve survival with a lasting immune-mediated control of GBM by using synthetic immunomodulatory gene circuits driving GBM cells to express a local combination of immunomodulatory proteins: human IL15, a surface T-cell engager, PD-L1-CD3 bispecific antibody, and the protein, LIGHT (TNFRSF14). Importantly, the co-expression of LIGHT and anti-PD-L1 therapies was recently shown to rescue PD-L1 checkpoint blockage in the preclinical models of brain tumors and significant enhance survival outcomes highlighting the benefits of novel combinations of immunomodulatory proteins for treatment of GBM. To identify genes whose expression is dramatically upregulated in GBM compared to normal human brain cells, a pooled of six thousand lentiviral oncogene promoters that drive expression of a red-fluorescent protein has been infected into three human GBM cell lines. RESULTS/ANTICIPATED RESULTS: We have successfully infected our GBM cells and are preparing samples for next generation DNA sequencing to determine highly active promoters in GBM that are not expressed in multiple normal brain cells types, astrocytes and neurons. These chosen promoters will then be used to drive an AND gate logic gene circuit immunotherapy outputs which is currently under development for both in-vitro and in-vivo experiments. DISCUSSION/SIGNIFICANCE OF IMPACT: We anticipate that local expression of multiple immune effectors proteins will significantly enhance tumor control and survival in both synergistic murine and human-murine xenograft pre-clinical models of GBM. Ultimately, our goal is to rapidly translate this technology advance into the clinical trial for adult GBM patients.

OBJECTIVES/SPECIFIC AIMS: Objective: Approximately 86 million people in the US have prediabetes, but only a fraction of them receive proven effective therapies to prevent diabetes. Further, the effectiveness of these therapies varies with individual risk of progression to diabetes. We estimated the value of targeting those individuals at highest diabetes risk for treatment, compared to treating all individuals meeting inclusion criteria for the Diabetes Prevention Program (DPP). METHODS/STUDY POPULATION: METHODS: Using a micro-simulation model, we estimated total lifetime costs and quality-adjusted life expectancy (QALE) for individuals receiving: (1) lifestyle intervention involving an intensive program focused on healthy diet and exercise, (2) metformin administration, or (3) no intervention. The model combines several components. First a Cox proportional hazards model predicted onset of diabetes from baseline characteristics for each pre-diabetic individual and yielded a probability distribution for each alternative. We derived this risk model from the Diabetes Prevention Program (DPP) clinical trial data and the follow-up study DPP-OS. The Michigan Diabetes Research Center Model for Diabetes then estimated costs and outcomes for individuals after diabetes diagnosis using standard of care diabetes treatment. Based on individual costs and QALE, we evaluated NMB of the two interventions at population and individual levels, stratified by risk quintiles for diabetes onset at 3 years. RESULTS/ANTICIPATED RESULTS: Results: Compared to usual care, lifestyle modification conferred positive benefits for all eligible individuals. Metformin’s NMB was negative for the lowest population risk quintile. By avoiding use among individuals who would not benefit, targeted administration of metformin conferred a benefit of $500-$800 per person, depending on duration of treatment effect. When treating only 20% of the population (e.g., due to capacity constraints), targeting conferred a NMB of $14,000-$18,000 per person for lifestyle modification and $16,000-$20,000 for metformin. DISCUSSION/SIGNIFICANCE OF IMPACT: Conclusions: Metformin confers value only among higher risk individuals, so targeting its use is worthwhile. While lifestyle modification confers value for all eligible individuals, prioritizing the intervention to high risk patients when capacity is constrained substantially increases societal benefits.

OBJECTIVES/SPECIFIC AIMS: Suggestibility, defined as the inclination to accept and internalize messages, has not been assessed much in relation to alcohol use. Prior research has shown that suggestibility to social cues and peer influence may play a role in driving alcohol consumption. Our previous work has shown associations between suggestibility and alcohol consumption in social drinkers. This study aims to examine how suggestibility and social susceptibility are related to ideas alcohol consumption and consequences across the spectrum of alcohol use and misuse. We hypothesize that those with higher suggestibility and social susceptibility reports will also have higher alcohol consumption and consequences, and that the impact of susceptibility is lower in dependent compared to non-dependent drinkers. METHODS/STUDY POPULATION: Study participants enrolled in the NIAAA screening and assessment protocol (N=157) completed questionnaires on suggestibility and alcohol consumption, The Multidimensional Iowa Suggestibility Scale (MISS) is a 95-question self-report assessment of suggestibility which draws from subcategories of consumer suggestibility, perceivability, physiological suggestibility, physiological reactivity, and peer conformity. Alcohol measures included 90-day Timeline Followback interviews and the Alcohol Use Disorder Identification Test (AUDIT). Participants also underwent the Structured Clinical Interviews for DSM-IV or DSM-5 disorders, and were stratified into two groups: alcohol dependent (N = 86) and non-dependent (N=71). Median split by age was additionally used to explore age’s relationship with suggestibility and alcohol with the under 36 (N = 45) and over 36 (N = 26) non-dependent groups. RESULTS/ANTICIPATED RESULTS: Initial analyses showed marked differences between the dependent and non-dependent groups in the relationship between the MISS total score and AUDIT total score. The non-dependent group showed significant positive correlations between MISS and AUDIT scores (r = 0.460, p < 0.001), while the dependent group did not show any correlation between MISS and AUDIT scores. Further examination of these relationships in the nondependent group revealed that MISS scores were also significantly positively correlated with AUDIT subscores of consumption, harm, and dependence. Age was found to have a significant negative correlation with MISS score (r = −0.354, p < 0.01). To better understand the role of age, the sample was split based on the median age (36 yrs), and analyzed separately. Results indicated robust relationships between MISS score and AUDIT (r = 0.457, p < 0.01) in the younger age group. In addition, the younger age group also showed significant relationships between MISS score and 90-day TLFB measures of total drinks, days drinking, and heavy drinking days. DISCUSSION/SIGNIFICANCE OF IMPACT: In non-dependent individuals, there was a significant positive relationship between suggestibility and alcohol measures, and these effects were amplified in younger individuals. No relationship was found between suggestibility and alcohol measures in the alcohol dependent individuals. This may be related to a greater impact of social and external contextual cues in younger social drinkers compared to dependent drinkers where tolerance and craving may have greater impact on alcohol consumption. These findings have important implications for traits that may put individuals at risk for developing harmful patterns of alcohol use and misuse. Future analyses will aim to examine biobehavioral underpinnings of the relationship between suggestibility and alcohol consumption.

OBJECTIVES/SPECIFIC AIMS: The goal of this project is to study fetal pulmonary vasculature in a CDH animal model, to understand how FETO affects developing vasculature, and to develop a modifiable fetal tracheal occlusive therapeutic device that avoids previously seen sequelae of FETO, like alveolar distension, decreased surfactant production, and decreased Type II Pneumocytes. The primary outcome is lung volume/kilogram. The secondary outcomes are contrast-enhanced ultrasound perfusion metrics (Time to Peak, Mean Transit Time, Wash-in Rate, Wash-in Perfusion Index), pulmonary vascular density, Lung Injury Histology Scores, and Lung Compliance upon ventilation. METHODS/STUDY POPULATION: Congenital diaphragmatic hernias will be modeled by surgical hernia creation via maternal laparotomy and hysterotomy at gestational age 72 - 74 days. The ewe will undergo a second laparotomy at 105 - 115 days gestational age. After a second hysterotomy is made, the fetus will be removed from the amniotic sac, though placental circulation will be maintained (EXIT Procedure). The animal is cannulated via the umbilical vein and arteries onto the pumpless ECMO circuit. The balloon and pressure sensor complex is placed into the trachea via direct laryngoscopy, and the fetus aseptically sealed into the Biobag. The wires of the tracheal occlusive device (balloon catheter and pressure sensor) will egress via the port of the Biobag. The fetus remains in the Biobag for fourteen days, with the tracheal occlusive device in place for ten days, followed by a four day recovery period. Daily contrast-enhanced ultrasounds and pulmonary artery dopplers are performed. Upon study completion, the fetus is intubated and placed on a conventional ventilator. A full necropsy is then performed, with perfusion fixation of the lungs via the pulmonary artery. RESULTS/ANTICIPATED RESULTS: Hypothesis 1: Modifiable Tracheal Occlusion will have statistically different effects on developing lung parenchyma, surfactant production, and abundance of Type II Pneumocytes Hypothesis 2: Modifiable Tracheal Occlusion will have lower levels of pulmonary hypertension than negative control animals, as measured by contrast-enhanced ultrasound (pulmonary artery velocity and washout time). DISCUSSION/SIGNIFICANCE OF IMPACT: This project will provide insight into the development of pulmonary hypertension in the CDH fetus. It will provide insight into the physiology of FETO, a novel therapy for congenital diaphragmatic hernias, and will demonstrate the utility of the EXTEND System for fetal treatments that are not possible in the maternal uterus.

OBJECTIVES/SPECIFIC AIMS: (1) To evaluate the association of patient and clinical factors with adherence to adjuvant hormone therapy (HT). (2) To examine the association of HT-related symptoms and the extent of remediation with early discontinuation of hormone therapy. METHODS/STUDY POPULATION: Retrospective cohort study of risk factors for interruption and early discontinuation of adjuvant hormone therapy in hormone receptor-positive nonmetastatic breast cancer patients diagnosed between 2009 and 2015. This study will include incident hormone receptor-positive breast cancer patients who initiated their HT and were followed at Tufts MC until Dec 31, 2016. Primary data source is electronic medical records (EMRs) RESULTS/ANTICIPATED RESULTS: The primary outcome of this study is early discontinuation to HT, defined as the first treatment gap of greater than or equal to 180 days following the initiation of HT. Treatment interruption, defined as any patient- or provider-initiated treatment gap of ≥ 2 weeks, will be examined as the secondary endpoint. Any HT-related symptoms occurred during a follow-up interval will be captured and categorized into five major types (i.e., vasomotor, neuropsychological, gastrointestinal, gynecological, and musculoskeletal symptoms). Onset and duration of a HT-related symptom will be recorded. Severity of the symptoms will also be rated by clinical oncologists. Remediations in response to HT- related symptoms will be collected and categorized into to two groups (pharmacological or non-pharmacological) and whether they were patient- or provider-initiated. Response to a remediation is defined as complete relief, partial relied, no relief, or with worsening symptoms. Response to a treatment change (i.e., HT switch or hold) was collected separately but using the same criteria. Analyses will be performed on the association between patient and clinical factors with rates of nonadherence (unplanned treatment interruption and/or early discontinuation) of hormone therapy, respectively. We also will explore whether patients with elevated symptoms and/or incomplete remediation will have earlier discontinuation of hormone therapy. DISCUSSION/SIGNIFICANCE OF IMPACT: Through formal chart review, we will establish a dataset that contains highly detailed information about treatment-emergent symptoms and remediations, which will enable us to quantitatively assess the impact of these treatment factors on adherence to hormone therapy for breast cancer. The in-depth analysis of risk factors associated with nonadherence to hormone therapy will inform development of interventions to improve cancer outcomes.

OBJECTIVES/SPECIFIC AIMS: Physical inactivity and mineral imbalances greatly contribute to morbidity and mortality in patients with end-stage renal disease (ESRD). Barriers for engaging in physical activity and adhering to the hemodialysis diet have been reported predominantly with white participants from countries other than Puerto Rico. Therefore, this study’s aims were to explore the barriers and facilitators that Hispanic adults with end-stage renal disease encountered for engaging in physical activity and adhering to the hemodialysis diet. METHODS/STUDY POPULATION: Three focus groups were conducted among 19 adults living with ESRD who received services from a renal center in Puerto Rico. Sessions were recorded, transcribed, and coded first using inductive methods. RESULTS/ANTICIPATED RESULTS: The presence of fatigue, lack of acceptance of the renal condition, and lack of knowledge of appropriate exercises for patients in hemodialysis were the most frequently reported barriers to engage in physical activities. Cost of the renal diet, limited availability of the renal diet products, the restrictive nature and the lack of Puerto Rican taste of the renal diet, and inadequate educational materials were the most frequent barriers to adhere to the hemodialysis diet reported by the sample. The most commonly reported facilitators to engaging in physical activities were having a positive attitude, opportunities for group exercises, and listening to Hispanic music while exercising. Health benefits, family support, having financial resources, availability of community resources, and having willpower were the most commonly reported facilitators to adhere to the hemodialysis diet. DISCUSSION/SIGNIFICANCE OF IMPACT: We identified a number of culturally relevant individual, interpersonal, institutional, and community-related barriers and facilitators to physical activity and adherence to the hemodialysis diet in patients with ESRD living in Puerto Rico. Evidence-based solutions to overcome these barriers and strategies for enhancing these facilitators should be addressed in future studies aimed at increasing the level of physical activity and increasing adherence to the hemodialysis diet in patients with ESRD living in Puerto Rico.

OBJECTIVES/SPECIFIC AIMS: The study aims to understand if pro inflammatory epicardial white adipose phenotype is positively associated with coronary atherosclerosis, while the brown adipose phenotype is negatively associated. Primary outcome is association between epicardial fat fraction and coronary atherosclerosis and cardiac function. Secondary outcome is transcriptomic and lipidomic profiling between epicardial, extra pericardial, and subcutaneous depots and how these profiles correlate with fat fraction. METHODS/STUDY POPULATION: Recruited patients undergoing open-heart surgery provided informed consent at their second visit and underwent laboratory testing and imaging (cardiac magnetic resonance including water-fat imaging and coronary calcium computed tomography) prior to their surgery. Cardiac function such as cardiac chamber volume, mass, function, and strain, and depo-specific fat fraction were calculated from cardiac MR and Agatston calcium score and epicardial adipose volume from CT images. At the time of surgery, a tissue specimens from the epicardial, extrapericardial, and subcutaneous depots were obtained for transcriptomic and lipidomic analysis. Linear and logistic regression analyses adjusted for other variables were performed to evaluate significance level between variables. RESULTS/ANTICIPATED RESULTS: 37 subjects were enrolled in the study, 13 (35%) of which were women. Cardiac function and fat fraction was quantified in all patients, whereas tissue analyses were performed in 22 patients. Epicardial and extrapericardial fat fraction were independently associated with coronary atherosclerosis (p-value 0.01 and 0.04 respectively) Only epicardial fat fraction was negatively associated with global circumferential shortening of the left ventricle (0.03), while neither the extrapericardial fat fraction nor epicardial adipose volume were not (p =0.33 and 0.97 respectively) All three adipose depots have unique gene signatures with differentially expressed genes and pathways. RNA sequencing of epicardial, extrapericardial, and subcutaneous depots demonstrated tight clustering of epicardial and subcutaneous signatures based on PCA analysis (Figure 2). 19 lipid classes and 59 lipids showed differential expression between at least 2 of the fat depots (Figure 3). Hierarchal clustering of the lipids showed that epicardial and extrapericardial depots were more closely related than subcutaneous adipose. Plasmenyl-phosphatidylcholines, with an ether-linked fatty acid at the sn-1 position of the lipid, were higher in subcutaneous fat while most other lipids were higher in epicardial fat per tissue weight, such as ceramides (p=0.002). DISCUSSION/SIGNIFICANCE OF IMPACT: Epicardial, extrapericardial, and subcutaneous adipose depots express different lipidome and transcriptome signatures and different pathways. GSEA analysis demonstrated enrichment of genes related to antigen presentation and B cell immunity in epicardial compared to subcutaneous adipose tissue. Epicardial fat fraction is associated with coronary atherosclerosis and decreased left ventricular global circumferential shortening as an early predictor of decreased left ventricular stroke volume. Epicardial fat fraction is also associated with cermides which may play role in the development of coronary atherosclerosis and decreased cardiac function.

OBJECTIVES/SPECIFIC AIMS: Type 2 diabetes (T2D) is costly and burdensome, but strong evidence exists that lifestyle change and weight loss can improve glycemic control and lower co-morbidities for patients with T2D. We used national data to examine whether the frequency of diet and/or physical activity counseling for patients with T2D in ambulatory settings has been responsive to accumulation of evidence supporting lifestyle change. METHODS/STUDY POPULATION: We used National Ambulatory Medical Care Survey (NAMCS) data over the period 2005-2015 from 31,475 patients with provider-reported T2D. We built multivariate logistic regression models, adjusting for patient, provider, and practice level characteristics (i.e. patient demographics, physician specialty, site of care, and region), to assess changes over time in the provision of diet or exercise counseling during ambulatory care visits, as reported through provider/staff chart review. We also examined whether changes in counseling over time varied by key patient and provider characteristics. We used non-overlapping confidence intervals (CI) to assess for statistical significance. RESULTS/ANTICIPATED RESULTS: Proportions of patients with T2D who received diet or exercise counseling were no different over time: 30% in 2005 [95% CI: 25%-35%] and 25% in 2015 [95% CI: 18%-31%]. Adjusted models show Hispanic patients had higher likelihood of receiving diet or exercise counseling, compared to whites (OR: 1.38 [CI: 1.03-1.85] for diet; OR: 1.37 [CI: 1.01-1.85] for exercise), and younger age was associated with higher likelihood of diet or exercise counselling, compared to those over 75 (age 30-49, OR: 1.47 [CI:1.18-1.82] for diet OR: 1.63 [CI: 1.30-2.03] for exercise). Among provider and practice-level characteristics, metro area and type of provider were associated with higher odds of receiving any diet and/or exercise counseling with visits in a metro area (OR: 1.23 [CI: 1.03-1.48]) and with an advanced practice provider (OR: 1.77 [CI: 0.97-3.22] having higher likelihood of receiving any diet or exercise counseling. DISCUSSION/SIGNIFICANCE OF IMPACT: Up to 30% of Americans with diabetes received any diet or exercise counseling in ambulatory visits, and this remained low over a decade. There were significant differences in counseling across patient, provider, and practice characteristics. Future studies are needed to better understand what interventions might improve counseling in ambulatory settings.

OBJECTIVES/SPECIFIC AIMS: To further explore the role of vWF in the pathogenesis of scleroderma by identifying its location within the tissue of sample biopsies obtained as part of routine diagnosis with the use of immuno-histochemical staining. METHODS/STUDY POPULATION: We examined 8 skin biopsies from 2 patients with systemic sclerosis (SSc), 2 with localized scleroderma (LS) and 4 with JDM. Double immunofluorescence staining was performed in each tissue with antibodies against vWF and collagens type I and III. DAPI (4′, 6-diamidino-2-phenylindole) was also used for counterstaining of inflammatory cells. Tissue staining patterns were compared between groups. RESULTS/ANTICIPATED RESULTS: Biopsies were obtained from the upper extremity of 7 females and the lower extremity of 1 male. Median age, symptom duration, and serum levels of vWF antigen around the time of biopsy was 8 years (IQR 4.5-11), 5.5 months (IQR 2.5-7), and 245% (IQR 203-302 for 7 patients), respectively. All but 1 biopsy was performed prior to initiation of immunosuppressive therapy. Immunofluorescence staining showed a superficial and deep perivascular inflammatory cell infiltrate that co-localized with vWF in all tissues. There was expression of vWF in the extravascular tissue of patients with JScl co-localizing with collagen III in the reticular dermis (Figures 1 and 2). In comparison, vWF expression was restricted to the endothelium and did not co-localize with collagen in the dermis of patients with JDM (Figure 3). Patients with SSc had higher expression of vWF as compared to patients with LS. DISCUSSION/SIGNIFICANCE OF IMPACT: vWF may participate in the pathogenesis of cutaneous inflammatory conditions. We have demonstrated that vWF co-localizes with cellular inflammatory infiltrates in the perivascular areas and in the dermis of patients with JScl and JDM. We additionally speculate that vWF may participate in the pathogenesis of fibrosing skin diseases based on evidence of increased extravascular expression in the tissue of patients with JScl (vs. JDM), and its co-localization with collagen. vWF expression intensity in the dermis of JScl patients may relate to disease extension (SSc vs. LS).