OBJECTIVES/GOALS: Strong evidence has implicated oxidative stress (OS) as a disease mechanism in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). The study aim was to assess whether a C>T single nucleotide polymorphism (SNP) (rs1800668), which reduces the activity of glutathione peroxidase 1 (GPX1), is associated with brain OS in patients with ME/CFS. METHODS/STUDY POPULATION: Study population: The study enrolled 20 patients with ME/CFS diagnosed according to Canadian Consensus Criteria, and 11 healthy control (HC) subjects. Genotyping: DNA was extracted from whole blood samples, amplified by PCR, and purified. Sanger sequencing was used for genotyping. 1H MRS: Proton magnetic resonance spectroscopy (1H MRS) was used to measure levels of glutathione (GSH)— a primary tissue antioxidant and OS marker— in a 3x3x2 cm3 occipital cortex (OCC) voxel. GSH spectra were recorded in 15 minutes with the standard J-editing technique. The resulting GSH peak area was normalized to tissue water level in the voxel. Statistical Analysis: T-tests were used to compare OCC GSH levels between ME/CFS and HC groups, and between the study’s genotype groups (group 1: CC, group 2: combined TC and TT). RESULTS/ANTICIPATED RESULTS: Clinical characteristics: ME/CFS and HC groups were comparable on age and BMI but not on sex (p = 0.038). Genotype frequencies: Genotype frequencies in the ME/CFS group were 0.55 (CC), 0.25 (TC) and 0.2 (TT); and 0.636 (CC), 0.364 (TC), and 0 (TT) in the HC group. GSH levels: There was a trend-level lower mean OCC GSH in ME/CFS than in HC (0.0015 vs 0.0017; p = 0.076). GSH levels by genotype group interaction: Within the ME/CFS group but not in the combined ME/CFS and HC group or HC group alone, GSH levels were lower in the TC and TT genotypes than in CC genotypes (0.00143 vs 0.00164; p = 0.018). DISCUSSION/SIGNIFICANCE: This study found that the presence of a C>T SNP in GPX1 is associated with lower mean GSH levels and, hence, brain oxidative stress, in ME/CFS patients. If validated in a larger cohort, this finding may support targeted antioxidant therapy based on their genotype as a potentially effective treatment for patients with ME/CFS.

OBJECTIVES/SPECIFIC AIMS: This presentation will highlight a structured, collaborative approach to implementing and utilizing the RPPR process created at the University of Minnesota CTSI in response to the need to enhance the quality, efficiency, consistency, and utilization of annual program reporting. The approach is in line with the NCATS’s strategic objective that encourages all CTS organizations to “disseminate research results and best practices broadly, and promote a culture of openness, sharing and transparency” (NCATS, 2016, p. 19). Program activities that support translational processes and contribute to clinical outcomes are complex, nonlinear, and multidisciplinary (Smith etal., 2017). In this complex context, the meaningful engagement and reflection of program staff and collaborators is essential for all aspects of program planning, implementation, reporting, and dissemination. The University of Minnesota CTSI’s key objectives, goals, and uses of RPPR are as follows: - Develop, align, and leverage the RPPR to fulfill the accountability requirements, needs, and expectations of multiple stakeholders: NIH/NCATS, Internal Advisory Board and External Advisory Board, campus/hub, program staff and collaborators. - Engage the CTSA staff and collaborators as a team in multiple aspects of program reporting. - Inform strategic management, continuous improvement, monitoring and evaluation, organizational learning and dissemination to program stakeholders. - Translate the reported information into practical, evidence-based issues and strategic questions for the leadership discussions and advisory board consultations, actionable work plans, communication to stakeholders, organizational learning, and translational science knowledge base. METHODS/STUDY POPULATION: A case study of the programmatic/evaluative and methodological approach/technique development that resulted in a formal, structured, collaborative, transparent process with detailed guidelines, templates, and timelines. The process and content for reporting has been developed via a variety of methods and sources: specific funder (NIH) requirements, Huddle meetings, document/content/database analysis, reflection meetings with component staff, informal conversations, and observations. Preparation for the report began almost one year in advance, including careful analysis of the report requirements, developing user-friendly, detailed guidelines, templates, and examples. The guide templates and worksheets were created as a result of time spent navigating current instructions provided by NIH and NCATS. Timeline/project plan was developed with start and end dates for all of the moving parts along with identified responsible personnel for each of the tasks. A grid of the grant components and responsible personnel was designed to highlight the matrixed organization of the grant and the need to work across components to create single reports. The RPPR key categories have also been considered for incorporating and tracking in a program activity/customer tracking system for ongoing data management and use. As a complex translational science program, UMN CTSI has multiple initiatives, variables, and metrics to report. The program staff has been deeply engaged in the evaluative reflection to identify, prioritize, and incorporate into the RPPR the metrics that most useful to manage and describe CTSI processes, participation, products, and outcomes. Program components responded differently to the collaborative approach implemented. The M&E technical assistance was implemented in 3 different ways: components either did the M&E RPPR template themselves, with minimal M&E team assistance; responded to comments and information provided by the M&E team as a first step; or requested a significant level of assistance from M&E. Participants/partners in developing and using RPPR include CTSI program leadership and staff, administration, communication staff, M&E team, and our collaborators. RESULTS/ANTICIPATED RESULTS: The proposed comprehensive approach to the annual program performance reporting shows sound promise to enhance program staff engagement, report utilization, learning, strategic management, self-evaluation capacity, and continuous improvement within a clinical and translational science organization. DISCUSSION/SIGNIFICANCE OF IMPACT: This structured approach’s impact is significant in that it fills the current gap in the practice, literature, and methodology and offers a practical example of a “practice that works” for CTR (and other) organizations and programs striving to improve their reporting practices, staff engagement, learning, and program impact. Leveraging and synergizing the RPPR requirements and other complex, data-demanding obligations and needs can help the CTS programs move beyond the once-a-year compilation of project accomplishments and challenges to developing and sharing a thoughtful translational science program success story. References: National Center for Advancing Translational Sciences. (2016). NCATS Strategic Plan. NIH. Available at: https://ncats.nih.gov/strategicplan Smith, C., Baveja, R., Grieb, T., & Mashour, G. (2017). Toward a science of translational science. Journal of Clinical and Translational Science, 1(4), 253-255. doi: 10.1017/cts.2017.14

OBJECTIVES/GOALS: Designed sampling from databases (DSD) methods have been used to cross-check electronic medical records for errors, structure study design, and, we hypothesize, can be used to make data collection for surgical quality metrics more efficient, particularly within national databases. We plan to apply statistical and DSD methods to accomplish the following aims:

1. 1. Identify the most important elements in managing post-operative pain

2. 2. Identify the most informative procedure or population-based targets to focus collection of additional, labor-intense detail surrounding adequacy of pain control (i.e., Patient Reported Outcome Measures (PROMs)).

METHODS/STUDY POPULATION: Our study population includes all children, ages 1-18 years, captured in the National Surgical Quality Improvement Project-Pediatric (NSQIP-P) from 2019 to 2021. We plan to apply statistical (regression modeling) and DSD methods to accomplish the aims listed above. RESULTS/ANTICIPATED RESULTS: For Aim 1, we expect to identify patient, procedure, and perioperative pain management practices that influence postoperative pain. For Aim 2, we will focus on outcomes such as PROMs that are challenging to obtain. By applying DSD methods, we will identify specific procedure and/or population-based cohorts to capture PROMs and decrease data collection burdens, while maintaining power, as the project is scaled nationally to all of NSQIP-P. DISCUSSION/SIGNIFICANCE OF IMPACT: Data from this study will inform expansion of NSQIP-P to collect novel outcomes of clinical and societal importance without prohibitively increasing data collection burden.

ABSTRACT IMPACT: Understanding the mechanism underlying the DASH diet may shed light on the physiologic process by which nutrition influences blood pressure and potentially lead the way to new therapeutics that target ion channels. OBJECTIVES/GOALS: Hypertension is a disease of the westernized world, as it stems from lifestyle habits. Lower salt consumption reduces blood pressure, yet DASH diet is much more effective, lowering blood pressure as efficiently as one anti-hypertensive drug. The precise mechanism through which DASH achieves its effect is not understood, and this is the project goal. METHODS/STUDY POPULATION: We hypothesize that exposing hypertensive volunteers to a high potassium and low sodium DASH diet will change the composition of renal ion channel in an aldosterone-dependent manner, leading to excretion of both sodium and potassium and a reduction in blood pressure. To assess how the nutritional change changes ion channel composition in the kidneys’ epithelium in aldosterone-induced manner, we will monitor urine exosomes, which contain epithelial cell membranes. We designed an in-hospital nutritional studies recruiting 20 volunteers. Patients will consume carefully designed menu, and measurements will be collected daily: blood pressure, biologic samples including blood and urine electrolytes, aldosterone, and urine for exosomes. RESULTS/ANTICIPATED RESULTS: We have collected data from 5 research volunteers so far.following exposure to the high potassium diet, Aldosterone blood levels increased while blood level of both potassium and sodium was maintained within normal limits. Urinary ratio of electrolytes, sodium:potassium was reversed 5-7 days following nutritional change from 6 to 1. Both manual and automated 24-hour blood pressure measurements confirmed blood pressure reduction following nutritional change. The following illustrates the impact the diet had on a participant’s 24-hour ambulatory blood pressure. Daily mean blood pressure was reduced from 120/76 mmHG to 112/68, mean awake blood pressure was reduced from 125/80 mmHG to 117/72 mmHG, and mean sleep blood pressure was reduced from 112/69 to 103/60 mmHG DISCUSSION/SIGNIFICANCE OF FINDINGS: Understanding the mechanism underlying the DASH diet may shed light on the physiologic process by which nutrition influences blood pressure and potentially lead the way to new therapeutics that target ion channels. By introducing our participants to a healthier lifestyle, they could maintain lower blood pressure without requiring medication.

Implementation of clinically useful research discoveries in the academic environment is challenged by limited funding for early phase proof-of-concept studies and inadequate expertise in product development and commercialization. To address these limitations, the National Institutes of Health (NIH) established the National Centers for Accelerated Innovations (NCAI) program in 2013. Three centers competed successfully for awards through this mechanism. Here, we present the experience of one such center, the Boston Biomedical Innovation Center (B-BIC), and demonstrate its remarkable success at the translation of innovations to clinical application and commercialization, as well as skills development and education.

OBJECTIVES/GOALS: In this study, we aim to report the role of porins and blaCTX-M β-lactamases among Escherichia coli and Klebsiella pneumoniae, focusing on emerging carbapenem resistant Enterobacterales (CRE) subtypes, including non-carbapenemase producing Enterobacterales (NCPE) and ertapenem-resistant but meropenem-susceptible (ErMs) strains. METHODS/STUDY POPULATION: Whole genome sequencing was conducted on 76 carbapenem-resistant isolates across 5 hospitals in San Antonio, U.S. Among these, NCP isolates accounted for the majority of CRE (41/76). Identification and antimicrobial susceptibility testing (AST) results were collected from the clinical charts. Repeat speciation was determined through whole genome sequencing (WGS) analysis and repeat AST, performed with microdilution or ETEST®. Minimum inhibitory concentrations (MIC) were consistent with Clinical and Laboratory Standards Institute (CLSI M100, ED33). WGS and qPCR were used to characterize the resistome of all clinical CRE subtypes, while western blotting and liquid chromatography with tandem mass spectrometry (LC-MS-MS) were used to determine porin expression and carbapenem hydrolysis, respectively. RESULTS/ANTICIPATED RESULTS: blaCTX-Mwas found to be most prevalent among NCP isolates (p = 0.02). LC-MS/MS analysis of carbapenem hydrolysis revealed that blaCTX-M-mediated carbapenem hydrolysis, indicating the need to reappraise the term, “non-carbapenemase (NCP)®” for quantitatively uncharacterized CRE strains harboring blaCTX-M. Susceptibility results showed that 56% of all NCPE isolates had an ErMs phenotype (NCPE vs. CPE, p < 0.001), with E. coli driving the phenotype (E. coli vs. K. pneumoniae, p < 0.001). ErMs strains carrying blaCTX-M, had 4-fold more copies of blaCTX-M than ceftriaxone-resistant but ertapenem-susceptible isolates (3.7 v. 0.9, p < 0.001). Immunoblot analysis demonstrated the absence of OmpC expression in NCP-ErMs E. coli, with 92% of strains lacking full contig coverage ofompC. DISCUSSION/SIGNIFICANCE: Overall, this work provides evidence of a collaborative effort between blaCTX-M and OmpC in NCP strains that confer resistance to ertapenem but not meropenem. Clinically, CRE subtypes are not readily appreciated, potentially leading to mismanagement of CRE infected patients. A greater focus on optimal treatments for CRE subtypes is needed.

OBJECTIVES/SPECIFIC AIMS: Of the six Centers for Medicare and Medicaid Services (CMS) monitored diagnoses targeted for readmissions reductions, reasons for readmissions within academic hospitals are poorly understood and reflect complex interactions between the patient, provider and organizational-level responses to initial hospitalization. Learning health systems (the organizational and orchestrated integration of research into evidence-based practice) can address the complexities of readmissions through an innovative approach to knowledge translation and patient-centered outcomes research. The objective of this review is to define and optimize the architecture of learning health systems to produce a dynamic pre-implementation framework of knowledge translation and patient-centered outcomes research, leveraging two engines (research and learning) within the academic and clinical settings for reducing readmissions. METHODS/STUDY POPULATION: Three databases were utilized for this scoping review (PubMed, Academic Search Premier, and Scopus) focusing on 1.) learning health systems and the methods of defining and building these systems within an academic hospital setting and 2.) the use of learning health systems in reducing readmissions within academic hospitals. Empirical articles and reviews pertaining to the architecture, development, conceptualization, definition, and translation of learning health systems were identified and compiled into a scoping review and proposed framework. RESULTS/ANTICIPATED RESULTS: The scoping review yielded 139 articles; from which 28 articles were retained. No articles were found utilizing learning health systems to address readmissions. Thus, a new architectural framework was developed incorporating common architectural themes from the literature with adaptations to fit the interests of patients, providers, and researchers in reducing readmissions within academic hospitals (Figure 1). DISCUSSION/SIGNIFICANCE OF IMPACT: Given the dearth of information applying learning health systems to readmissions, the proposed architecture for an integrative learning health system can be utilized as a dynamic foundation for adoption and pre-implementation planning for reducing readmissions within academic hospital settings. Additionally, the authors expect this model to be tested and continually refined to address historical and emerging issues for clinically-relevant and clinically-effective approaches to patient-centered practice and research.

ABSTRACT IMPACT: Currently, there are no medications to treat metabolic syndrome and our research sheds light on a potential therapeutic that could prove beneficial for this disease that affects one-third of the US population. OBJECTIVES/GOALS: Our goal was to determine the role of the GI tract in MetS, specifically how approved GI-directed medications affect metabolic parameters. Thus, we assessed the effects of mesalamine, a common therapeutic utilized to treat mild to moderate UC, on metabolic parameters in comorbid UC and MetS patients METHODS/STUDY POPULATION: This was a retrospective study with data extracted from Cerner’s HealthFacts database across the United States (US). Inclusion criteria included adult patients (≥18 years old) with a diagnosis of UC and at least 3 of the 5 metabolic risk factors which included i) dyslipidemia, ii) low HDL, iii) hyperglycemia, iv) hypertension, and v) increased abdominal obesity as determined by elevated BMI. A total of 6197 patients across the US between the years of 2007 and 2017 were included. We pulled patients who had a mesalamine prescription within +/- 7 days of an encounter in which they were diagnosed with UC (index date) and the closest values to 3 and 12 months after the index date. Mean age for patients was 53.8 ±19.9, with predominance of female sex (52.9%) and white race (78.0%). RESULTS/ANTICIPATED RESULTS: There was an observed reduction in BMI, fasting glucose, and increase in HDL levels post start of mesalamine treatment along with a decrease in inflammatory markers (ESR and CRP) (p<0.001). DISCUSSION/SIGNIFICANCE OF FINDINGS: The GI tract contributes to numerous disorders associated with metabolic dysfunction. Our retrospective analysis revealed mesalamine treatment in comorbid UC and MetS patients improved metabolic parameters, providing evidence that targeting the GI tract in these individuals potentially improves dysregulated metabolic processes.

ABSTRACT IMPACT: A better understanding of the spectrum of problematic opioid use will lead to more targeted treatments. OBJECTIVES/GOALS: It is unclear how to approach treatment of individuals with problematic opioid use who do not clearly meet criteria for opioid use disorder (OUD). We aim to characterize clinical, demographic, and medication use at time of identification of problematic opioid use across the spectrum as well as identify predictors of poor outcomes. METHODS/STUDY POPULATION: A national sample of Veterans coded as having opioid abuse or dependence were previously categorized as (1) high likelihood of OUD, (2) limited aberrant opioid use, and (3) prescribed opioid use with no evidence of aberrant use based on chart review. We will describe how individuals in these three categories differ demographically and clinically. We will then use a trained binary logistic regression model to predict whether individuals with limited aberrant opioid use more closely resemble category (1) or (3). Cox proportional hazards models will be used to predict all-cause mortality, suicide-related mortality, opioid-overdose related mortality, and hospitalization over a three-year period using the three categories as predictors and adjusting for relevant covariates. RESULTS/ANTICIPATED RESULTS: We anticipate that Veterans with a high likelihood of OUD will be more likely to experience homelessness and have more psychiatric comorbidities (especially PTSD). We hypothesize that Veterans with prescribed opioid use and no evidence of misuse will be significantly older, more likely to have disability, medical comorbidities (ie., chronic pain, cancer), more prescriptions for non-opioid analgesics, and be prescribed higher doses of opioids. Using a trained binary logistic regression model, we predict that Veterans with limited aberrant opioid use will more closely resemble Veterans with a high likelihood of OUD. We expect that all categories of problematic opioid use will have a high risk of mortality, with a high likelihood of OUD associated with the greatest risk of premature death. DISCUSSION/SIGNIFICANCE OF FINDINGS: Identifying and better characterizing individuals with limited aberrant opioid use may be an important opportunity to intervene prior to development of severe OUD. Future research will focus on targeting interventions to this population, which may have specific needs that are separate from classic OUD or simple pain-related opioid dependence.

OBJECTIVES/GOALS: Financial toxicity describes the adverse effects of medical expenses on financial security and health related quality of life. Though dementia caregiving carries serious costs, financial toxicity has not been studied in this context. Here we assess the prevalence of financial toxicity in dementia caregiving and its sociodemographic correlates. METHODS/STUDY POPULATION: We utilized the COmprehensive Score for financial Toxicity (COST)— 12-item questionnaire validated to quantify financial toxicity in patients and their caregivers— to conduct a nationally representative survey of 317 US dementia caregivers, oversampling non-Hispanic Black (n = 75) and Hispanic (n = 61) caregivers. Participants were required to be currently providing unpaid care to someone 50 years or older with dementia. Financial toxicity was defined as COST 0 & RESULTS/ANTICIPATED RESULTS: COST scores ranged between 0 and 44, with a survey-weighted mean of 24.57 and standard deviation of 9.8. Weighted analysis revealed 52.7% of American dementia caregivers experience some degree of financial toxicity. Of those who experience financial toxicity, 73.1% are classified as mild, 25.7% as moderate, and 1.2% as severe. Financial toxicity was identified in 69.5% of non-Hispanic Black, 54.1% of Hispanic, and 42.3% of non-Hispanic White caregivers, with non-Hispanic Black caregivers significantly more likely to experience financial toxicity compared to their non-Hispanic White counterparts (p = 0.017). DISCUSSION/SIGNIFICANCE: Most US dementia caregivers experience financial toxicity, though prevalence varies significantly by caregiver race. Discerning the pervasiveness of financial toxicity in this population and significant correlates will inform the development and expedient delivery of resources for patients and families.

OBJECTIVES/GOALS: The aim was to examine whether nicotine patch was more effective in encouraging abstinence from cigarettes smoking compared to placebo. METHODS/STUDY POPULATION: Randomized controlled trials involving the general teenage age group smokers who were current smokers-“smoked less than 100 cigarettes over their lifetime and smoked at the time of the interview. Databases were searched for relevant studies reported in English that employed a randomized design published since 2000. Two authors extracted data and assessed quality. The primary outcomes and prioritization were continuous abstinence at 3, 6 and 12-month follow-up or more for the number of patients who responded to treatment, defined as a reduction/abstinence. Heterogeneity between studies did not preclude combined analyses of the data. RESULTS/ANTICIPATED RESULTS: 4 of 266 publications were included. Four studies reported positive effects on smoking cessation at end of treatment: (1) nicotine patches improved continuous abstinence at 6 weeks – 9 weeks months; (2) nicotine patch improved continuous abstinence at 3 to 6 months; (3) nicotine patches improved continuous abstinence 6 and 12 months; (4) nicotine patches improved continuous abstinence at 6 months – 12 and 24 months (5). All studies showed, continuous abstinence at follow up differed in percentage between groups both at 6 weeks through 24 months, with NRT (Nicotine patch) intervention groups achieving higher rates in most of the studies compared to placebo intervention group. Conclusions: NRT intervention methods seem to increase smoking abstinence in those treated for smoking cessation. Further and larger sample size studies are required to make stronger the base of evidence. DISCUSSION/SIGNIFICANCE OF IMPACT: Four randomized controlled trials investigating the effectiveness of smoking cessation interventions, for teenagers who smoke cigarettes were identified for inclusion in this review. Four of the studies reported significant effects on smoking cessation, providing evidence of effectiveness of NRT (nicotine patch), behavioral support and combinations of the two, although not all trials intervention treatments found an effect. The four studies reported important intervention effects at both the short and long follow-ups required: 6 weeks up to the 24 months, thereby, providing stronger evidence to support the effectiveness of NRT intervention on smoking cessation. All studies showed some evidence of improved smoking abstinence outcomes. The four studies had in common that the smoking cessation interventions provided a combination of intent to treat prevention, and of all the clinical trials none of them suggested a negative effect of smoking cessation treatment on substance use outcomes using NRT. However, the studies used reliable methods and reported their cases properly, but the small number of studies reviewed for the systematic review makes the conclusion about the effectiveness of these interventions uncertain. The papers visibly stated how the trials protected against bias, as indicated by the Yes (low risk). No (high risk) and U as “unclear risk.” All four studies conducted a random sequence generation of participants enrolled into the study sample.

OBJECTIVES/GOALS: Evaluate the impact of COVID-19 on oral clefts services including surgical and dental treatments in Puerto Rico. METHODS/STUDY POPULATION: This Observational retrospective cohort study will consider patients 0-21 y/o with CL/P that visited the UPR school of Dental Medicine, Pediatric University Hospital Dr. Antonio Ortiz and ongoing case-control research project Face-Genes. Records to be used are classified as follow: Pandemic (March 15, 2020 to March 15 2022) Pre-pandemic (March 15, 2015 to March 15, 2017) Power analysis (power=0.80 alpha=0.05) will be calculated. Unavailable and incomplete medical records and those that did not attended study clinic during study period will be excluded. Data extraction instrument will be based on previous published study. Descriptive statistics, Chi-square, Odds Ratios at 95% confidence intervals and multiple logistic regression will be estimated. RESULTS/ANTICIPATED RESULTS: We hypothesize that surgical and dental services in Puerto Rico will be adversely impacted because of COVID-19 pandemic. DISCUSSION/SIGNIFICANCE: CL/P are common congenital diseases that require early interdisciplinary attention. Lack of timely care as well as surgery and treatment delays, could be associated with poorer prognosis, increased morbidity and mortality. If there is high risk of dh services during emergency situations, our findings will help to allocate the available resources

ABSTRACT IMPACT: Our data identify a novel candidate for combination strategy in melanoma treatment, and can inform clinicians in their decision-making process regarding therapeutic intervention for melanoma patients. OBJECTIVES/GOALS: Soluble adenylyl cyclase (sAC) is a novel source of cyclic AMP (cAMP). In melanoma, nuclear sAC localization has an established diagnostic utility and we newly found that nuclear sAC functions as a tumor suppressor by inhibiting Hippo pathway, which affects treatment response. Here, we examine the effect of nuclear sAC on melanoma treatment response. METHODS/STUDY POPULATION: We developed a doxycycline inducible system for increasing sAC activity only in the nucleus. We assessed whether nuclear sAC activity affects treatment response, using BRAFV600 human melanoma cell lines. Using a clonogenic assay, we examined how nuclear sAC activity affects growth inhibition in the presence of a BRAF inhibitor, vemurafenib. Our findings will be confirmed in vivo using tumor xenografts. After tumor formation in NSG mice, mice will be randomized to be fed normal or doxycycline chow for nuclear sAC induction, then subdivided to receive vehicle or vemurafenib to examine the effect of nuclear sAC activity on treatment response in vivo. We will also compare melanoma biopsies collected before and after treatment with BRAF inhibitors to assess how nuclear sAC staining affects tumor morphology in vivo. RESULTS/ANTICIPATED RESULTS: So far, nuclear sAC activity has rendered SkMel178 and M263 cell lines more susceptible to vemurafenib. Cell viability was inversely correlated both with vemurafenib and with doxycycline concentration. Cell viability after vemurafenib treatment was dramatically reduced when nuclear sAC was activated. It appears that nuclear sAC enhances the sensitivity of BRAF mutant melanomas to vemurafenib in vitro. We anticipate that xenografts of these cells in mice will be more susceptible to vemurafenib when nuclear sAC is activated. We also anticipate that positive nuclear sAC staining will correlate with a favorable response to therapy. DISCUSSION/SIGNIFICANCE OF FINDINGS: Targeted therapy with BRAF inhibitors is used in late-stage melanomas, but its use is limited as patients invariably acquire resistance. Here, we identified nuclear sAC activation as a novel candidate for combination strategy. Our data will also inform clinicians how best to integrate this biomarker into their decision-making regarding therapy.

OBJECTIVES/SPECIFIC AIMS: The objective of this exploratory study is to evaluate the relationship between the individual genetic variants in COMTrs4680 and DRDrs1076560 and relevant alcohol use behaviors (i.e. alcohol consumption and reward processing behaviors) in non-dependent drinkers within experimentally controlled IV-ASA CAIS sessions. The overall goal of this study is to begin gathering data on the influence of individual genetic variants on alcohol consumption and other drinking-related behaviors. This will aid in the creation of a polygenic model of risk for AUD which will provide more insight into how the mesolimbic pathway is affected by alcohol use. METHODS/STUDY POPULATION: Study population: The sample included male and female non-dependent drinkers (N=149). Genotypes for functional polymorphisms in COMT (rs4680) and DRD2 (rs1076560) genes were determined for all subjects from blood samples obtained during screening. Alcohol consumption was assessed using the 90-day Timeline Followback Interviews (TLFB). Study population demographics: Self-reported gender (53.5% identified as male); Self-reported race (61.2% identified as white); Age ranged from 21-46 years old, with 22 years being the mode. Experiment: Free access (open-bar) intravenous alcohol self-administration (IV-ASA) using the computer-assisted alcohol infusion system (CAIS) paradigm; Subjects had the choice of pressing a button ad libitum for IV alcohol infusions during the session, neurobehavioral questionnaires were collected throughout the 2.5-hr alcohol infusion session. Primary outcome measures included: Total Rewards, Peak breath alcohol concentration (BrAC) achieved, and Total Ethanol consumed. Statistical Analyses: Conducted using SPSS IBM Statistics Versions 1.0.0-2482; non-dependent drinkers were organized into two groups based on their genotypes, minor allele carriers and major allele homozygotes. Outcome measures were compared between genotype groups using analysis of variance or non-parametric Mann-Whitney U-test as appropriate. RESULTS/ANTICIPATED RESULTS: -We expect the genetic makeup of the sample to be reflective of larger genome samples that are publically available (e.g. e!Ensembl) - Initial analysis for COMTrs4680 did not reveal significant effects on IV-ASA measures. Specifically, the majo DISCUSSION/SIGNIFICANCE OF IMPACT: Alcohol Use Disorder (AUD) affects millions of men and women globally. The heterogeneity within AUD individuals has made it difficult to identify biological and/or psychological factors that could be targeted for the development of treatments. By using the human laboratory model of free access IV-ASA, this study evaluated the relationship between dopaminergic genetic variants, COMTrs4680 and DRDrs1076560, and alcohol consumption in non-dependent drinkers within a controlled experimental environment. This study will begin to evaluate genetic and behavioral data that can be used to create a polygenic model of risk for AUD, which will provide more insight as to how the mesolimbic reward pathway is affected by alcohol use and contributes to risk for AUD.

OBJECTIVES/GOALS: COVID-19 disproportionately affects patients with prior health conditions and those living at a lower socioeconomic status. Persons living with HIV (PLWH) are infected with SARS-CoV-2 at a higher rate than seronegative patients. Risk factors and incidence of post-COVID-19 comorbidities in PLWH, specifically, are still unknown METHODS/STUDY POPULATION: We will study PLWH enrolled in the Emory Centers for AIDS Research (CFAR) Registry who receive care at the Grady Ponce de Leon Center in Atlanta, Georgia to 1) investigate the incidence of, and 2) identify risk factors that predispose PLWH to post-COVID-19 comorbidities. All PLWH with documented COVID-19 (by positive SARS-CoV-2 PCR or antigen test) between March 1, 2020, and September 30, 2021, with a clinic visit within 12 months will be included. We will identify comorbidities using problem list diagnoses and ICD9/10 codes. With a predicted sample size of 395, we will use a Cox proportional hazards model for time-to-detection of comorbidity, and bivariate and multivariate logistic regression models to identify predictors of incident comorbidity within 12 months of COVID-19. RESULTS/ANTICIPATED RESULTS: o Previous work demonstrated that in PLWH, age and non-AIDS comorbidities, but not HIV-related factors, were associated with hospitalization for COVID-19 in a dose dependent fashion.18 We anticipate that rate of incident comorbidities will be significantly higher in PLWH after COVID-19 compared to PLWH without a history of COVID-19. We also expect that pre-existing comorbidities including obesity and cardiovascular disease, male sex, Black race, and older age are associated with higher incidence of post-COVID-19 comorbidities in PLWH. When stratifying by organ system, we also anticipate that prior comorbidities of an organ system will predispose patients to later complications of that same system. DISCUSSION/SIGNIFICANCE: By understanding the incidence and risk factors associated with developing post-COVID-19 comorbiditieswe can improve guidelines for treatment of groups experiencing the disproportionate impact of co-infection with HIV and SARS-CoV-2.

OBJECTIVES/GOALS: Leveraging Patient’s Experience to improve Diagnosis (LEAPED) is our proposed method of measuring diagnostic error through seeking patient feedback on their understanding of their diagnosis and health status following emergency department discharge. To pilot test LEAPED’s feasibility, we deployed and determined patient uptake of LEAPED. METHODS/STUDY POPULATION: To test LEAPED, we employed a longitudinal cohort study design at emergency departments across one academic health system in the Mid-Atlantic region. Patients consented to complete questionnaires regarding their understanding of their diagnosis and/or follow-up steps and their health status at 2 weeks, 1 month, and 3 months following emergency department discharge. People aged 18 and older who were seen at the emergency department within the past 7 days with at least one chronic condition (hypertension, diabetes, history of stroke, arthritis, cancer, heart disease, osteoporosis, depression, and/or chronic obstructive lung disease) and one or more of the following common chief complaints: chest pain, upper back pain, abdominal pain, shortness of breath/cough, dizziness, and headache were eligible to join the study. RESULTS/ANTICIPATED RESULTS: Of those enrolled (n = 59), 95% (n = 53) responded to the two week post-ED discharge questionnaire (1 and 3-month ongoing). Of the 6 non-responders, 1 had died and 3 were hospitalized at two weeks. The average age was 50 years (SD 16) and 64% were female. Over half of participants (53%) were white and 41% were black. Almost one-third (27%) reported they were not given an explanation of their health problem on leaving the ED, and of those, a third did not have an understanding of what steps to take after leaving the ED. Participants reported a new health problem was identified after ED discharge (19%), worsening health status (12%), and health status stayed the same (16%). DISCUSSION/SIGNIFICANCE OF IMPACT: Patient uptake of LEAPED was high, which suggests that patient-report is a feasible method of evaluating diagnostic decision making and delivery to patients and yields insightful information beyond administrative data. The next steps are to validate the accuracy of patient-reported diagnostic error by comparing with administrative data.

OBJECTIVES/GOALS: Using biomarkers to identify vulnerabilities from cocaine use disorder (CUD) is a focus of recent investigations. Current clinical efforts focus on psychiatric recovery in CUD, however other body systems are missed. Applying blood-based transcriptomics to investigate how clinical conditions relate to CUD can alter current treatment approaches. METHODS/STUDY POPULATION: We conducted a comprehensive longitudinal study of 12 individuals (mean 53 yrs.; M/F ratio 9: 3) with CUD abstinent from cocaine. 44 blood samples collected repeatedly every 3 months for 9 months were bulk RNA sequenced. We began with phenotype harmonization grouping individuals with the following metrics; cocaine withdrawal, cue craving, generalized craving, perceived stress, and days of abstinence. We ran differential gene and transcript expression with time across grouping of metrics using the dream software and used the multivariate test to examine their associations. We used the association of gene-transcripts to determine genetic predispositions with clinical traits using the Multi-marker Analysis of GenoMic Annotation assessing their overlap to a reference GWAS database. RESULTS/ANTICIPATED RESULTS: Individuals were grouped in 2 clusters based on scores of cue craving, generalized craving, cocaine withdrawal, and 3 clusters based on days of abstinence from cocaine use. Gene-transcript(s) associationsrevealed genetic predisposition towards certain clinical conditions and substance use traits. Cannabis use disorder showed significant enrichment between the greater vs. lesser abstinent days, and lesser vs. least abstinent days at 9 months. The “drinks per week” trait showed significant gene enrichment between greater vs. lesser abstinent days at 9 months. Coronary artery disease was also enriched with greater vs. least abstinent days at 3 months. Lastly, significant baseline differences in predisposition to small vessel ischemic stroke were seen in responders with high vs low perceived stress. DISCUSSION/SIGNIFICANCE: These results from a robust and feasible pipeline suggest genetic predisposition in CUD for other substances and cardio-neurovascular diseases. This pilot study may lead to larger studies into whole genome-based blood biomarker approaches for monitoring abstinence and other clinical co-morbidities to be addressed in cocaine addiction recovery.