OBJECTIVES/GOALS: The objective of this study is to define the molecular mechanisms that control survival of malignant stem cells in acute myeloid leukemia (AML). Leukemia stem cells (LSCs) are not effectively eradicated by standard treatment and lead to resistance and relapse, which contribute to poor survival rates. METHODS/STUDY POPULATION: The recently FDA approved venetoclax, a BCL2 inhibitor, with azacitidine, a hypomethylating agent leads to a 70% response rate in AML patients. Analysis of patients treated with this regimen showed direct targeting of LSCs. BCL2 has a non-canonical function in regulation of intracellular calcium. To determine how BCL2 mediated calcium signaling plays a role in LSC biology, we used LSCs isolated from venetoclax/azacitidine (ven/aza) sensitive and resistant patient samples to measure expression of calcium channels via RNA seq. BIO-ID, siRNA, flow cytometry, seahorse assays, calcium measurements and colony assays were used to determine the effects of calcium channel perturbation on LSC biology. RESULTS/ANTICIPATED RESULTS: BCL2 inhibition leads to decreased OXPHOS activity in primary AML specimens. BIO-ID studies revealed cation/metal ion transporters, ER membrane proteins and ER membrane organization as top enriched pathways interacting with BCL2. RNA-seq data showed increased expression of genes involved in calcium influx into the ER in ven/aza sensitive LSCs and increased expression of genes involved in calcium efflux from the ER in ven/aza resistant samples. Ven/Aza resistant LSCs have increased mitochondrial calcium content, consistent with their increased OXPHOS activity as calcium is required for OXPHOS. Perturbation of these channels leads to decreased OXPHOS activity and decreased viability in LSCs. DISCUSSION/SIGNIFICANCE OF IMPACT: We postulate that a deeper understanding of the mechanisms behind ven/aza targeting of LSCs will lead to the development of novel therapies for patients who do not respond to ven/aza. Our data show targeting intracellular calcium signaling could be a viable therapeutic strategy for AML patients.

Objectives/Goals: Gait performance (GP) and global cognitive functions (GCF) are both critical for maintaining independence and quality of life in senior adults. Recent studies have explored the potential link between GP and GCF, encompassing executive functions. Methods/Study Population: PRISMA guidelines will govern this systematic review. This systematic review synthesizes published research from 2000 to 2024, including peer-reviewed articles, pilot studies, and randomized controlled trials, to examine the relationship between GP (how a person walks and stands) and GCP in older adults. The exclusion criteria will be based on studies focused on physical activities unrelated to balance, meta-analysis, and systematic reviews and those published in languages other than English or Spanish. Results/Anticipated Results: Our preliminary data indicate that gait, or walking speed, is significantly correlated with GCP in older adults, with slower walking associated with poorer global cognition. Specifically, gait speed during dual-task walking shows a strong correlation with working memory (p < .001) and processing speed (p < .05) in individuals aged over 60 years. Gender differences were observed, with women over 80 walking slower than men over 85 years, who walked faster, and women exhibited poorer global cognition than men. Discussion/Significance of Impact: Overall, a gait slowing between 0.68 and 1.1 m per second could predict a marker for the risk of developing dementia, indicating that monitoring gait speed in older adults may provide early warning signs, allowing for timely interventions. Enhancing GP and GCF can improve the quality of life and independence in older adults. Acknowledgments: Research supported by NIH: Award Number HCTRECD R25MD007607 from the NIMHD.

OBJECTIVES/GOALS: Identification of COVID-19 patients at risk for deterioration following discharge from the emergency department (ED) remains a clinical challenge. Our objective was to develop a prediction model that identifies COVID-19 patients at risk for return and hospital admission within 30 days of ED discharge. METHODS/STUDY POPULATION: We performed a retrospective cohort study of discharged adult ED patients (n = 7,529) with SARS-CoV-2 infection from 116 unique hospitals contributing to the national REgistry of suspected COVID-19 in EmeRgency care (RECOVER). The primary outcome was return hospital admission within 30 days. Models were developed using Classification and Regression Tree (CART), Gradient Boosted Machine (GBM), Random Forest (RF), and least absolute shrinkage and selection (LASSO) approaches. RESULTS/ANTICIPATED RESULTS: Among COVID-19 patients discharged from the ED on their index encounter, 571 (7.6%) returned for hospital admission within 30 days. The machine learning (ML) models (GBM, RF,: and LASSO) performed similarly. The RF model yielded a test AUC of 0.74 (95% confidence interval [CI] 0.71–0.78) with a sensitivity of 0.46 (0.39-0.54) and specificity of 0.84 (0.82-0.85). Predictive variables including: lowest oxygen saturation, temperature; or history of hypertension,: diabetes, hyperlipidemia, or obesity, were common to all ML models. DISCUSSION/SIGNIFICANCE: A predictive model identifying adult ED patients with COVID-19 at risk for return hospital admission within 30 days is feasible. Ensemble/boot-strapped classification methods outperform the single tree CART method. Future efforts may focus on the application of ML models in the hospital setting to optimize allocation of follow up resources.

ABSTRACT IMPACT: This work has the potential to identify targetable pathways conveying resistance to PARP inhibitors that may improve ovarian cancer patient outcomes. OBJECTIVES/GOALS: High grade serous ovarian cancer is the deadliest gynecologic malignancy. PARP inhibitors are an FDA approved targeted therapy that is being used more and more frequently in the clinic. It is vital to understand mechanisms driving resistance to this therapy in order to develop treatments to improve patient responses. METHODS/STUDY POPULATION: RNA-sequencing and transcription factor analysis was used to identify pathways of interest. An AP-1 transcriptional reporter assays was used to confirm results of the transcription factor analysis. An unbiased lentiviral shRNA screen was used to identify AP-1 subunits promoting PARP inhibitor resistance. Lentiviral transduction allowed for the knockdown ATF6. Comet assays and two-plasmid systems were used to determine levels of DNA damage and levels of DNA damage repair respectively. RESULTS/ANTICIPATED RESULTS: PARP inhibitor resistant cell lines have increased WNT signaling which promotes to increased DNA damage repair. PARP inhibitor resistant cell lines also have increased AP-1 transcriptional activity, ATF6 expression, and active p38. ATF6 knockdown and p38 inhibition is sufficient to resensitize cells to PARP inhibition. Upon treatment with PARP inhibitors, ATF6 knockdown as well as p38 inhibition lead to increased DNA damage in PARP inhibitor resistant cell lines. RNA-sequencing reveals a signifcant overlap in downregulated genes in cells treated with a β-catenin inhibitor and cells with an ATF6 knockdown. DISCUSSION/SIGNIFICANCE OF FINDINGS: Due to the increasing prevalence of PARP inhibitors in the clinic, it is vital to uncover mechanisms contributing to resistance. This work has the potential to identify targetable pathways conveying resistance to PARP inhibitors that may improve ovarian cancer patient outcomes.

OBJECTIVES/GOALS: Clinical trials of SGLT2 inhibitors in patients with heart failure (HF) have confirmed a reduction in hospitalization and death. Adoption of novel therapeutics has been slower in Black and female patients. We investigated utilization of SGLT2 inhibitor in patients with HF and type 2 diabetes and if there were utilization differences by race or gender. METHODS/STUDY POPULATION: We created a retrospective cohort of outpatients with HF at Emory Healthcare from 2015 to 2020. Additional inclusion criteria included presence of heart failure and a diagnosis of T2D. SGLT2 inhibitor use was identified by a presence of SGLT2 inhibitor prescription at the time of the clinic visit. We estimated differences in prescription of SGLT2 inhibitors by race and gender using Chi-square analysis. RESULTS/ANTICIPATED RESULTS: The cohort included 5829 patients, age 69.47 years ± 13.44, 47.67 % female, 54.62% Black. Overall prescription of SGLT2 inhibitors was low but increased over time (1.4% in 2015 to 5.6% in 2020; p<0.0001). On average, SGLT2 inhibitor use increased annually by 44.77%. From 2015 to 2020, fewer female than male patients were on an SGLT2 inhibitor (1.94% vs. 2.73%, p=0.0033). A similar percentage of Black and non-Black patients were on an SGLT2 inhibitor (2.13% vs. 2.64%, p=0.0591). DISCUSSION/SIGNIFICANCE: Prescription rates of SGLT2 inhibitors remain low in patients with T2D and HF, especially for female patients, despite evidence of their benefit on hospitalizations and mortality. Implementing use of SGLT2 inhibitors in this population represent an opportunity to improve cardiovascular outcomes.

OBJECTIVES/GOALS: Bronchiolitis is a major cause of PICU admission, yet identifying best practices is limited by the lack of existing databases containing the needed demographic and clinical variables. We used probabilistic linkage to join the Virtual Pediatric System (VPS) and Pediatric Health Information Systems (PHIS) databases to overcome this data barrier. METHODS/STUDY POPULATION: We performed a single site study joining VPS and PHIS data. These national databases contain clinical (VPS) and billing and resource use (PHIS) data. Limits on the use of patient identifiers (PI) for multi-center research, makes direct linkage techniques impossible. To demonstrate that probabilistic linkage can accurately link VPS and PHIS records, we obtained our single site VPS and PHIS records and linked them using probabilistic linkage without PI and compared this to a gold standard linkage created with PI. We also compared demographic features of linked and unlinked records to assess the ability of probabilistic linkage to create a representative sample. RESULTS/ANTICIPATED RESULTS: We obtained 920 VPS records of patients with bronchiolitis and linked 91% (839/920) to a PHIS record with 4 (0.5%) false-positive matches. Characteristics of linked and unlinked records are compared in Table 1. Comparison of probabilistically linked and unlinked records showed no difference in median age in years (0.7 [Interquartile range (IQR) 0.3-1.5] v 0.7 [IQR 0.2-1.5], p = 0.76), median number of complex chronic conditions (0 [IQR 0-1] v 0 [IQR 0-1], p = 0.16), median Pediatric Index of Mortality 3 severity of illness scores (-4.6 [IQR -4.7 – 4.4] v -4.6 [IQR -4.7 – 4.4], p = 0.44), median days of PICU stay (4 [IQR 3-6] v 4 [IQR 2-6], p = 0.36), proportion female ( 44% v 46%, p = 0.82), or proportion of patients intubated (28% v 24%, p = 0.41). DISCUSSION/SIGNIFICANCE: Probabilistic linkage creates an accurate combined VPS-PHIS database. Extending our methodology to join data from all 38 hospitals contributing to VPS and PHIS will allow creation of a large database containing the demographic, treatment, and outcome data needed to enable Comparative Effectiveness Research of bronchiolitis care.

OBJECTIVES/SPECIFIC AIMS: The objective of this partnership was to create a global network of clinical and public health researchers and communities conducting technology-assisted research in noncommunicable disease. METHODS/STUDY POPULATION: The University of Rochester’s Clinical and Translational Science Institute (CTSI) has successfully leveraged the informatics core’s capacity into an emerging network of organizations that focus on technology and health in settings outside of the mainland United States. The CTSI coordinated with another NIH-funded infrastructure program [the RCMI Translational Research Network (RTRN)] to identify partner institutions interested in technology and health. RTRN identified the University of Puerto Rico and the University of Hawaii, both of which serve as hubs for common research interests in technology and health throughout the Caribbean and the Pacific. This network was formalized as the CDC’s Coordinating Center for its Global and Territorial Health Research Network (the “Global Network”), with additional US partners (Yale, University of Illinois at Chicago, University of North Caroline Chapel Hill, and the University of South Florida) within a wider scope of the CDC’s Prevention Research Centers (PRC) program. RESULTS/ANTICIPATED RESULTS: Through combining 2 main NIH-funded research infrastructure networks (CTSA and RTRN), with a large CDC-funded PRC, the University of Rochester’s Informatics Core was successful in establishing a new productive global health network throughout Latin America and the Caribbean, and in the Pacific, garnering additional research support from NIH Fogarty and other programs. The resulting network not only supports locally-important research in technology and health on compelling health issues (eg, diabetes, ZIka, participation in research), but also facilitates community engagement through local partnerships and the cores of the involved networks. In addition, much of the information and communications technology (ICT)-related research and learnings from the Global Network activity is immediately applicable to populations in the United States, served by the various collaborative networks. In total, while new, the Global Network supports a wide range of projects and engagements throughout the world that expand local informatics capacity and use of technology in the research process and to address global health problems, further enhancing the CTSI’s informatics core to serve the needs of its own constituency and promote research engagement with technology within this population. Local research collaborative projects reinforce the utility of the network and its resources, evidenced by tools, publications, partnerships, and conference presentations that have arisen. Lessons to date from this Global Network collaboration include: specific global research projects provide opportunities for partnership building and meaningful collaboration, team science is of central importance in distributing the work of the network, synergy is multidirectional with expertise and need flowing in all directions, and project team members in all locales learned and contributed substantially in ways that carried into their other responsibilities. DISCUSSION/SIGNIFICANCE OF IMPACT: The overall partnership has created opportunity for South-South collaboration, for adaptation of projects among locales, and has helped boost reputational value for all partners involved. Implications for other CTSA awardees include: global collaboration can serve core research and technical needs for the CTSA itself and its local partners, CTSA status can be leveraged to access resources to support local research, and collaboration in other federally-funded research networks helps expand the insight, scope, and potential for new research.

Objectives/Goals: Unsafe sleep practices contribute to sleep-related infant mortality, a leading cause of preventable sudden unexplained infant death (SUID). Infant illness represents a risk for SUID. However, the mechanism behind this increased risk is unknown. The objective of this study was to measure changes to safe sleep practices during infant illness. Methods/Study Population: We performed a prospective cohort survey study of infants aged 0–12 months presenting to the pediatric emergency department. We assessed sleep practices prior to illness, during illness, and 2 weeks and 1 month following illness. We assessed adherence to American Academy of Pediatrics safe sleep recommendations at each point in time. Wilcoxon sign rank test was used to examine changes between time points. Regression models compared caregivers who reported a change to unsafe sleep practices with those who did not. Results/Anticipated Results: Of enrolled participants (n = 142), 110 (77%) completed all three follow-up surveys. For those with complete follow-up, 62.4% were female, 60.3% were Black, non-Hispanic, 25.7% were White, non-Hispanic. The most common chief complaint was respiratory illness (35.7%), followed by fever (22.7%), and 70.3% of patients were discharged home. Across all sleep behaviors surveyed, caregivers reported, on average, a 12% change to unsafe sleep practices during illness. These changes were sustained at the 2-week and 1-month follow-ups. Factors associated with a change to unsafe sleep practice were parental age. Discussion/Significance of Impact: Over 10% of infants experienced a change to unsafe sleep practices during illness, sustained at 2-week and 1-month follow-ups. This may explain the association of infant illness with SUID. Interventions promoting safe sleep adherence during illness are key to decreasing sleep-related infant mortality.

Objectives/Goals: In neurological animal research, statistical misapplication may lead to overoptimism in a therapy’s potential for successful translation. This pilot study investigated whether human clinical trials that fail have higher prevalence of statistical misapplication in preceding animal experiments, compared to human trials that succeed. Methods/Study Population: Phase 2 clinical trials for 3 neurological conditions were identified on ClinicalTrials.gov and classified as successful or failed based on advancement to Phase 3 and/or preplanned efficacy test results. PRISMA guideline methods were used to systematically search for preclinical animal experiments (same indication and intervention) preceding the start of the human trial. Data were gathered from animal articles by collectors blinded to human trial outcome and included items describing reporting transparency, experimental design and sample sizes, and statistical tests applied. Statistical mistakes were coded based on mismatch between test and design. Rates of mistakes were compared between articles preceding successful and non-successful human trials using weighted point estimates and 95% confidence interval. Results/Anticipated Results: The final sample included 24 trials (16 successful) and 70 associated animal studies. Transparency was poor, with infrequent reporting of group allocation method (39%), sample sizes adequate to evaluate attrition (Discussion/Significance of Impact: Statistical misapplication is common in animal research, and this pilot study has demonstrated that preclinical statistical mistakes may indeed occur more frequently prior to failed human trials. Mistakes and lack of transparency may lead to overoptimism in preclinical experimental findings, with consequences for subsequent human translation.

OBJECTIVES/SPECIFIC AIMS: We have reported that radiofrequency renal denervation (RF-RDN) in SHR at 20-weeks of age, decreased blood pressure (BP) and fibrosis in kidney cortex and medulla when rats were sacrificed at 6 months. However, whether RF-RDN can have similar benefits in older rats remains unknown. This study examined whether performing RF-RDN in older rats also has a beneficial effect on BP and renal fibrosis. METHODS/STUDY POPULATION: Baseline systolic and diastolic BP (SBP/DPB) was measured (telemetry) in nine-month-old SHR and Wistar Kyoto rats (WKY). Groups of rats then received bilateral RF-RDN or Sham-RDN (SHR-RDN, n=9; SHR-Sham, n=10; WKY-RDN, n=5; WKY-Sham, n=8). Rats were then sacrificed at 12-months of age. Kidneys were harvested, sectioned, and assessed for fibrosis by Masson’s trichrome stain. A pathologist, who was blinded to treatment groups, evaluated each kidney section for fibrosis. RESULTS/ANTICIPATED RESULTS: Compared to SHR with Sham-RDN, RF-RDN prevented a further increase in systolic and diastolic BP from baseline (9-month) in SHR as they aged to 12-months (SHR-Sham mmHg: 9-month 193±4/127±4; 12-month 207±3/142±5; SHR-RDN mmHg: 9-month 197±3/132±2; 12-month 197±4/132±3). RF-RDN did not alter SBP or DBP in aged WKY. One-year-old SHR with prior Sham-RDN showed extensive renal fibrosis in kidney cortex and medulla. In contrast, RF-RDN significantly decreased renal fibrosis in the medulla, but not cortex. There was no fibrosis in kidneys of age matched WKY. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings suggest that RF-RDN may be a potential therapy for halting progression of hypertension and decreasing medullary fibrosis in the aged population.

OBJECTIVES/GOALS: 1. Establish the prevalence of amenorrhea among women Ebola survivors of reproductive age. 2. Determine whether amenorrhea in Ebola survivors is associated with hyperthyroidism, primary ovarian failure, and/or low weight METHODS/STUDY POPULATION: This study will enroll a cohort of 150 women Ebola survivors and 150 uninfected controls from Kenema, Sierra Leone. Participants will complete women’s health questionnaires with detailed menstruation history and provide blood samples. Serum levels of thyroid stimulating hormone, follicle stimulating hormone, and albumin will be measured in order to explore the role of thyroid dysregulation, ovarian failure, and low weight in amenorrhea associated with prior Ebola infection. RESULTS/ANTICIPATED RESULTS: Enrollment for the study is still in progress. As of November 14, 2022, 68 female Ebola survivors of reproductive age and 124 uninfected controls have been enrolled. Among this preliminary group, there is a high baseline level of menstrual irregularities in both survivors and controls. Prior to the Ebola outbreak, 97% of all participants reported less than 6 periods per year and 59% reported periods lasting 3 days or less. The prevalence of missed periods increased after the Ebola outbreak in both groups. Four Ebola survivors (5.9%) reported missing periods before infection, compared to 16 (23.5%) survivors after infection. Two uninfected controls (1.6%) reported missing periods before the Ebola outbreak, compared to 12 (9.7%) after the outbreak. DISCUSSION/SIGNIFICANCE: The next steps of this project are to complete enrollment, conduct data analysis, and perform laboratory studies. An enhanced understanding of amenorrhea is needed to develop novel medical interventions, to inform healthcare guidelines and policies, and to develop personalized treatment strategies to better care for women who have survived Ebola.

ABSTRACT IMPACT: Novel adipokines like tetranectin help explain why some people progress from obesity to diseases like diabetes, atherosclerosis, and dislipidemia OBJECTIVES/GOALS: Obesity has an established association with diabetes, dyslipidemia, and atherosclerosis. Preventing progression from obesity to insulin resistance requires understanding of the regulatory mechanisms involved in the loss of insulin sensitivity. Adipose tissue is well known to function as an endocrine organ that produces many kinds of adipokines. METHODS/STUDY POPULATION: Blood sample analysis from human patients and mice was used to determine associations between tetranectin and obesity. Samples were tested with a monoclonal anti-tetranectin antibody for detection with western blot. A tetranectin mutant knock out mouse line was compared to wild type littermates on high fat diet for 4 months. Insulin tolerance tests and glucose tolerance were used to determine progression to insulin resistance and glucose intolerance. Histological analysis of metabolic tissue was used to demonstrate adipocyte hypertrophy and liver steatosis. RESULTS/ANTICIPATED RESULTS: In the current study, we report the identification and initial characterization of a novel adipokine tetranectin. Tetranectin, which is coded by the C-type lectin domain family 3 member B (CLEC3B) gene, is ubiquitously expressed in various mouse tissues, whereas it is highly enriched in white adipose tissue. We found that the serum level of tetranectin was much higher in both obese and diabetic patients. Knocking out the tetranectin gene in mice protected against glucose intolerance in males but reduced insulin and glucose tolerance in females, without effects on food intake and body weight for either sex. Mechanistically, tetranectin targets liver tissues and its deficiency increases lipid accumulation in hepatocytes in females. DISCUSSION/SIGNIFICANCE OF FINDINGS: We have identified a novel adipokine which mediates a different metabolic crosstalk among tissues to maintain systemic glucose and lipid metabolism in different genders. Further investigation of tetranectin’s function could yield a new target for precise therapeutic treatment for obesity and its associated metabolic diseases in different genders

OBJECTIVES/GOALS: To describe the impact of Trial-CARE (Coordination, Acceleration, and Recruitment Enhancement), a centralized service core at Washington University (WU), aimed at enhancing the capability of investigator-initiated multi-site clinical trials at WU and partner CTSA institutions. METHODS/STUDY POPULATION: Through review of our marketing materials and service tracking system we defined the following Trial-CARE offerings: Infrastructure: Trial-CARE is a centralized service core at WU. Whereby, a team member is sourced to an investigator-initiated multi-site clinical trial (II-MCT) to ensure the achievement of study milestones across any phase of the clinical trial life cycle. Team: Our team consists of research professionals with expertise in II-MCT project management, data management, and administrative/regulatory management. Services: * One free 60-minute case consultation * Tiers of Service * Academic Research Organization (ARO) support * Clinical Coordinating Center * Data Coordinating Center * II-MCT project management support * Time-limited targeted support RESULTS/ANTICIPATED RESULTS: Trial-CARE has completed 94 consultations in support of WU and partner CTSA institutions enabling the streamlining of study start-up; guidance for study recruitment, implementation, and operations; and offering resources to foster career development. Consultations can be completed at any phase in the clinical trial lifecycle, with Trial-CARE completing: * 12% in the idea phase * 45% in the grant development and submission stage * 28% after funding has been awarded Top reasons researchers connect with Trial-CARE about II-MCTs: * regulatory guidance (40%) * general information about Trial-CARE Services and II-MCTs (35%) * protocol development (21%) * data management (20%) * study budgeting (20%) DISCUSSION/SIGNIFICANCE: In response to a WU wide survey, Trial-CARE plans to generate informational webinars and is creating a clinical trial tracking dashboard, to pro-actively offer support services to researchers prior to grant funding. The goal is also to increase the percentage of consultations completed in the idea phase and the grant development and submission stage.

OBJECTIVES/SPECIFIC AIMS: To learn the edit distance costs of a symbolic univariate time series representation through a stochastic finite-state transducer to predict patient outcomes in intensive care units. METHODS/STUDY POPULATION: High frequency data of patients in intensive care units were used as a data set. The nearest neighbor method with edit distance costs (learned by the FST) were used to classify the patient status within an hour after 10 hours of data. Several experiments were developed to estimate the parameters that better fit the model regarding the prediction metrics. RESULTS/ANTICIPATED RESULTS: Different metrics were obtained for the several parameters. These metrics were metrics (ie, accuracy, precision, and F-measure). DISCUSSION/SIGNIFICANCE OF IMPACT: Our best results are compared with published works, where most of the metrics (ie, accuracy, precision, and F-measure) were improved.

OBJECTIVES/GOALS: Social anhedonia is considered a key feature of schizophrenia that leads to social withdrawal. Research in general anhedonia suggests those with schizophrenia exhibit deficits in anticipated (predicted), but intact consummatory (in-the-moment), pleasure. This study will determine if these temporal differences apply to daily social anhedonia. METHODS/STUDY POPULATION: This project will use experience sampling methods (ESM) to measure real-world social pleasure in people with schizophrenia and healthy controls. Using electronic surveys, participants will predict social activities they plan to do throughout their day and report anticipated pleasure for each. Then, in subsequent surveys, participants will report their consummatory social pleasure shortly after engaging in each of these activities. We will use a time-lagged approach to match anticipated/consummatory pleasure ratings for specific social events and compute a difference score to determine discrepancy. Multi-level modeling will be used to determine if clinical status (schizophrenia/control) predicts anticipated pleasure, consummatory pleasure, and/or their discrepancy. RESULTS/ANTICIPATED RESULTS: Data collection for this project was recently completed. The schizophrenia and control groups (n = 30 per group) were demographically matched for age, sex, race, and ethnicity. Data processing and analyses are currently underway. We hypothesize that, in line with laboratory research in general anhedonia, those with schizophrenia will exhibit deficits in anticipated, but not consummatory, social pleasure throughout their daily lives. Moreover, when anticipated and consummatory pleasure ratings for the same social activities are matched, we expect those with schizophrenia to exhibit larger discrepancies between predicted and in-the-moment pleasure. DISCUSSION/SIGNIFICANCE: This project serves as a critical intermediate step to bridge the gap between laboratory research and patient treatment. ESM bypasses limitations of laboratory studies and increases ecological validity. Results will provide a nuanced understanding of social anhedonia and help identify precise targets to treat social withdrawal in schizophrenia.

OBJECTIVES/SPECIFIC AIMS: These studies seek to evaluate hormonal regulation of luminal breast cancer lipid metabolism and to identify targetable progesterone-mediated changes in lipid biology that contribute to therapeutic resistance in breast cancer. METHODS/STUDY POPULATION: Established and patient-derived luminal breast cancer cell lines, which express ER and PR, were used for this study. RNA transcript and protein expression levels were evaluated by qRT-PCR and immunoblot, respectively. Broad scale lipidomics of progesterone-treated cells was conducted via ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS) through the UCD Skaggs School of Pharmacy Mass Spectrometry Core. RESULTS/ANTICIPATED RESULTS: Data mining of previously published microarray data of CK5+ and CK5− syngeneic cancer sublines revealed that CK5+ cells have increased expression of lipid processing genes, including LPL and PPARG. As progestin treatment induces a subpopulation of cells to turn on CK5 expression in luminal breast cancers, UHPLC-MS-based lipidomics analysis will expose whether modulation of the lipid landscape occurs in all cells with progesterone treatment, or whether this phenomenon is heightened specifically in CK5+ cells. I also expect that ER+ breast cancers with progestin induced-altered lipid content, such as lipid droplet formation, will evade therapy-induced death. DISCUSSION/SIGNIFICANCE OF IMPACT: There are numerous approved and developmental therapeutics targeting lipid biology. By determining if progestins alter lipid metabolic genes specifically in CK5+ CSCs, which are endocrine resistant, strategies may be devised to target these resistant cells using combination therapy in conjunction with existing therapies to prevent tumor recurrence.

OBJECTIVES/SPECIFIC AIMS: The goal of this study is to assess how quality of life scores change in menopausal women before and after implementation of this aid. In addition, we are also interested in 2 process evaluation objectives: (1) determine if MyChart, the patient portal, is an effective way for this patient population to provide insight their quality of life to their providers and (2) to evaluate providers use of and reactions to the decision support tool. METHODS/STUDY POPULATION: This project is a collaboration between University of Rochester Medical Center and S.U.N.Y. Upstate Medical University. Participants were recruited through Upstate’s Family Medicine and OB/GYN practices via a MyChart invitation sent by the practices. Participating patients will be asked to complete a survey, through MyChart, every 3 months for 18 months. Participating health providers will be trained to use the decision support tool and participate in 3 interviews with the researchers to gain insight into the usefulness and effectiveness of the tool. RESULTS/ANTICIPATED RESULTS: Of the 465 eligible women, 117 women responded to our MyChart invitation to join our study. Of these, 105 agreed to participate and 98 met eligibility criteria. Only half of the women currently enrolled in our study had spoken to a provider about menopause related symptoms (56.1%) prior to study enrollment. DISCUSSION/SIGNIFICANCE OF IMPACT: The goal of this study is to improve menopause related symptoms in women, thus increasing their quality of life, but it will also provide important process evaluation for using EPIC and MyChart for future research studies.

OBJECTIVES/GOALS: Research in Non Alcoholic Fatty Liver Disease Genetics is ongoing. In this project, we aim to: 1.Identify and describe a histological NAFLD Cohort in the MVP Biobank by extracting liver biopsy proven NAFLD patients using natural language processing (NLP) 2.Confirm previously defined NALFD genetic loci (via look up-GWAS) in the histological NLP cohort METHODS/STUDY POPULATION: We will utilize the Million Veteran Program Biobank where a total of 10,959 subjects have been identified using liver biopsy reports in the EMR via CPT codes. Cases will be based on i) steatosis, steatohepatitis, inflammation or fibrosis in liver biopsy reports with ii) exclusion of other causes of liver disease. Controls will be comprised of the general VA population. In collaboration with the Applied NLP and Precision Medicine groups at the VA Informatics and Computing Infrastructure(VINCI); we will attempt to create and validate a histological cohort of NAFLD in the MVP database by: 1)Annotation of biopsy reports for the NLP algorithm 2)Automation/training of the NLP algorithm We will then perform a multi-ancestry genetic lookup of previously established genetic loci among the cases identified. RESULTS/ANTICIPATED RESULTS: Recently published data: the MVP NAFLD research led by Dr. Chang and Dr. Vujkovic had first validated a proxy NAFLD phenotype based on chronic alanine aminotransferase elevation (cALT). A GWAS was then performed using this phenotype which revealed 77 loci of genome-wide significance including 10 established NAFLD- and 52 ALT-associated SNPs were identified. Replication in external Liver Biopsy and Imaging Cohorts validated 17 SNPs of which 9 were novel. Preliminary data on Liver Biopsy reports: Using CPT codes for Liver Biopsy we estimated a total of 10,959 unique patients and 18,812 notes in the MVP biobank. We anticipate 2,000-3,000 NAFLD cases (based on the prevalence of NAFLD in the general population). Initial review reveals 90% concordance between analysts for the purposes of developing an NLP. DISCUSSION/SIGNIFICANCE: NAFLD is a major cause for morbidity and mortality in liver disease. Gold standard for diagnosis is based on biopsy. GWAS studies with biopsy proven phenotypes are limited. The study will aim to isolate a histologically defined NAFLD cohort in MVP to conduct further GWAS that can provide new clinically relevant knowledge for future research.

Contemporary Research Challenges

OBJECTIVES/GOALS: Genetic variation in L-type voltage-gated calcium channels, including CaV1.3, is associated with increased risk for psychiatric disorders including bipolar disorder and schizophrenia. Additionally, rare mutations in CaV1.3 have been linked to epilepsy, developmental delay, and autism. Deletion of CaV1.3 in mice is associated with impaired consolidation of contextual fear conditioning. Some studies have also observed affective behavior deficits in CaV1.3-deficient mice, but other studies have not found affective phenotypes, perhaps due to differences in genetic backgrounds, sex ratios, or task protocols. CaV1.3 is important for slow afterhyperpolarization in hippocampal and amygdala neurons, which prevents excessive firing in response to sustained excitatory input, and CaV1.3-deficient amygdala neurons exhibit hyperexcitability and impaired LTP. CaV1.3 is also expressed in the cerebellum, but its functional role there is not well understood. Given its importance in shaping neuronal activity in the hippocampus and amygdala, we hypothesized that loss of CaV1.3 would cause abnormalities in motor learning as well as affective and cognitive behaviors. METHODS/STUDY POPULATION: Wild-type (WT), haploinsufficient (Hap), and knockout (KO) mice were maintained on a congenic C57BL/6NTac genetic background and were subjected to behavioral tasks including open field, rotarod, ErasmusLadder, elevated zero maze, forced swim test, and tail suspension test. Data were analyzed with sexes combined and with sexes separated to assess for sex as a biological variable. Studies were analyzed by one-way ANOVA, two-way ANOVA, or generalized linear mixed model, where appropriate. RESULTS/ANTICIPATED RESULTS: CaV1.3 KO was associated with impaired motor learning in the rotarod task (p < 0.05), as well as impaired associative learning in the ErasmusLadder task (p < 0.01), despite intact locomotor function on both tasks. When examined by sex, the rotarod phenotypes were driven by motor learning impairments in males (both Hap and KO, p < 0.05 and p < 0.01, respectively), whereas the ErasmusLadder associative learning phenotypes were present in both sexes only in the KO condition, consistent with previously reported impairments in CaV1.3-deficient mice in consolidation of contextual fear conditioning. Although KO mice learned the motor aspects of the ErasmusLadder task, they learned more slowly. They also failed to learn start cues, which requires intact associative learning. No differences were observed in overall exploration or locomotor activity in open field or elevated zero maze. Analyses from affective tasks are ongoing. DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary studies provide new evidence that CaV1.3 is important for the function of neural circuits involved in motor learning, and concur with previous data showing its involvement in associative learning. Our data differ slightly from previous studies of CaV1.3 in motor learning, which could be attributable to differences in task protocols and/or genetic background. These results highlight the importance of CaV1.3 in a variety of behaviors, which may help explain why variation in CaV1.3 expression and function has pleiotropic effects in humans.

OBJECTIVES/SPECIFIC AIMS: The objective of this study was to determine if dopamine signaling is altered in a mouse model displaying excessive self-grooming and further elucidate the potential utility of compounds targeting the striatal DA system in modulating repetitive behaviors. METHODS/STUDY POPULATION: Here, we report studies using fast-scan cyclic voltammetry (FSCV) in mice lacking the postsynaptic protein SAP90/PSD95-associated protein (SAPAP3 KO mice) as well as control littermates. Rodent self-grooming provides a behavioral output with which one can monitor repetitive, self-directed, patterned behavior that has great translational value to OCD-like disorders. Total time spent grooming was monitored in SAPAP3KO mice and control littermates. To further examine the role of DA in regulating repetitive grooming behaviors the magnitude and kinetics of DA transients were assessed using FSCV in ex vivo slice preparations as well as in anesthetized mice in vivo. DA transients were elicited in the dorsolateral striatum (DLS), dorsomedial striatum (DMS); and nucelus accumbens core (NAcc). In some experiments mice were crossed with DAT-Cre animals and channelrhodopsin 2 (ChR2) was virally expressed in DA neurons to allow optical stimulation of DA transients. RESULTS/ANTICIPATED RESULTS: As previously reported, SAPAP3 KO mice showed excessive grooming compared to control littermates at the age assessed (4-5 months). DA transients evoked by a single electrical pulse in slices from SAPAP3 KO mice were not significantly different from those observed in slices from control littermates in any of the regions tested including the DLS, DMS and NAcc. However, when four electrical pulses were applied at a frequency of 10Hz to mimic DA neuron bursting, the magnitude of DA transients observed in the DMS and NAcc of SAPAP3 mice were greater than those evoked in control littermates.Interestingly, phasic stimulation produced similar DA transients in the DLS of both genotypes suggesting that phasic DA signaling was not globally altered. To confirm this finding we crossed SAPAP3 KO mice with DAT-Cre mice and injected ChR2 containing virus into the midbrain to selectively express ChR2 in DA neurons. Transients were then optically evoked resulting in selective activation of DA neurons. Optical stimulation produced a pronounced enhancement of DA release in SAPAP3 KO mice specifically in the DMS and only following phasic-like stimulation. DISCUSSION/SIGNIFICANCE OF IMPACT: These exciting findings suggest that DA signaling in SAPAP3KO mice is dysregulated in a very precise manner that is sub-region specific as well as dependent on the pattern of stimulation. These results suggest that targeted therapies that can modulate these specific modes of dopaminergic signaling in these distinct striatal subregions could provide improved efficacy in OCD patients that are resistant to SSRI treatment.