OBJECTIVES/GOALS: Research ethics services are critical to the clinical, research, and educational missions of an academic health science center. Our ethics program aims to develop a culture where investigators are as intellectually engaged in ethical issues of scientific integrity as they are in study design, data collection, and implementation. METHODS/STUDY POPULATION: This descriptive analysis depicts the historical development, from 2010 to 2022, of our research ethics program as an exemplar of ethics integration into the research enterprise of an academic health science center that engages in translational research. In this culture, clinicians, translational researchers and their scientific peers, research participants, and community members become involved in ethics investigation, deliberation, and innovation. RESULTS/ANTICIPATED RESULTS: There are four pillars to our research ethics program: 1) research ethics consultation service, which fosters the development of ethical best practices and standards for the practice of translational research; 2) education, which provides customized training and educational opportunities in research ethics to diverse stakeholders; 3) leadership, through collaboration and partnerships; 4) scholarly engagement, in the pursuit of innovative ethics research and professional development. Through these initiatives we can engage a broad constituency of stakeholders, become an integral component of research oversight, engage as active participants in the research enterprise, and have a critical role in guiding institutional culture. DISCUSSION/SIGNIFICANCE: The integration of our ethics program mirrors the translational science continuum which promotes the multidirectional flow of ideas among ethics consultants, laboratory/clinical scientists, implementation researchers and the community.

OBJECTIVES/GOALS: Inexpensive, accurate home monitoring is the standard-of-care in many diseases like hypertension or diabetes; however, it has yet to be widely used in neurodegenerative diseases. We used wearable activity monitors and computer-vision evaluated assessments to estimate Parkinson’s disease (PD)-related disease burden. METHODS/STUDY POPULATION: We recruited 22 people from the University of Iowa Movement Disorders Clinic. Each person completed a standardized set of 3 fine motor tasks using their hands. We recorded a video of this activity, which was evaluated using MediaPipe - an open-source pose classification program from Alphabet - as well as had an nurse-practitioner evaluate the performance on a validated scale (UPDRS). Participants wore a Fitbit Inspire 3 activity tracker at home for the next two weeks. We quantified disease burden using the Parkinson’s Disease Questionnaire 39 - a validated 39-item survey about the intensity of PD-related impairment. Using data from the videos and activity trackers, we estimated 1) the standardized UPDRS assessment of motor impairment and 2) the total PDQ-39 score. RESULTS/ANTICIPATED RESULTS: We found observationally recorded fastest sustained (at least 5 minutes) walking speed was a strong predictor of PDQ-39, explaining over one third of the variability in the measure. Range of motion in the videos was a significant predictor of UPDRS scores; however, was only weakly related to the overall PDQ-39 score. Further processing of the signals from the video, including wavelets and frequency domain analysis, may provide better predictive capabilities. PDQ-39 subscores (e.g., cognition, social support, mobility) will be the subject of further analysis. DISCUSSION/SIGNIFICANCE: Home monitoring has become the standard in other fields because of the better generalizability of home measurements. Improving the detection and evaluation of PD using home monitoring will lead to more timely and accurately changes in medication and less need for clinic visits - especially off levodopa.

OBJECTIVES/GOALS: This study evaluates the utility of self-reported quality of life measure in children with hearing loss. We will compare self-reported HEAR-QL scores with parent-reported HEAR-QL scores. We will then test the relationship between HEAR-QL scores and scores on a standardized assessment of cognition, the NIH Cognition Battery. METHODS/STUDY POPULATION: We will administer the HEAR-QL questionnaire to children with hearing loss and their parents. We will then administer the NIH Cognition Battery to the child. We will include in our population children ages 7 to 14 with hearing loss of any severity or side. We will exclude those with intellectual disability, disorders of speech or language, or those who would be unable to complete the questionnaires for any reason. Children will be recruited from Otolaryngology clinics at St. Louis Children’s Hospital based on ICD diagnosis of sensorineural hearing loss between 01/2015 – 03/2020. RESULTS/ANTICIPATED RESULTS: We will aim to recruit 44 patients in total, which is the sample size needed to detect a moderate correlation (r = 0.4) with a 1-sided α = 0.05 and 1-β = 0.8. HEAR-QL scores and NIH Cognition Battery scores will be reported using descriptive statistics. Linear regression as well as correlation analysis between HEAR-QL scores and cognitive testing scores will be performed using a 1-sided α = 0.05, with 1-β = 0.8. If recruitment is sufficient, we will adjust for demographics that are significantly correlated with the outcome on multivariate analysis. Finally, we will test for agreement between parent report and child report by calculating a Kappa statistic. DISCUSSION/SIGNIFICANCE OF IMPACT: There is little clarity on the necessity of amplification in children with hearing loss, yet the child’s perspective is not routinely assessed in clinical practice. This study employs self-report in a pediatric population with hearing loss to find out if children provide new and reliable information.

OBJECTIVES/GOALS: This study was performed to explore the capabilities of simultaneous [89 Zr]trastuzumab-PET/MRI acquisition in a cohort of metastatic HER2+ breast cancer. The insights derived provide additional noninvasive characterization and precise intratumoral analysis tools for healthcare providers. METHODS/STUDY POPULATION: A total of 13 patients, aged between 40 and 70, diagnosed with HER2-positive breast cancer, were selected to participate in this study. Whole-body [89 Zr]trastuzumab-PET/MR imaging was performed 5 ± 1 days post-injection of the radiopharmaceutical during ongoing HER2-directed therapy. Concurrently acquired T1-weighted MRI facilitated the identification of normal organ and tumor regions of interest, which were further analyzed for mean ADC and mean standardized uptake value. Multiparametric intratumoral habitat analysis was performed. Utilizing the median metric values, tumors were evaluated for heterogeneity, specifically assessing high and low HER2 expression through an image processing framework in conjunction with ADC metrics. Long-term treatment response evaluation is ongoing. RESULTS/ANTICIPATED RESULTS: Initial analysis indicate all tumors exhibited higher overall uptake of [89 Zr]trastuzumab across various sites including the bone (p=0.019), brain (p=0.014), and breast (p=0.069), when compared to corresponding normal organs. Additionally, increased ADCmean values were observed in all regions besides brain tumors (bone: p=0.002, brain: p=0.5, breast: p=0.03, juxtapulmonary: p=0.037), indicating distinct patterns of cellularity. Notably, one of five patients with a breast lesion, who exhibited a complete response to HER2-targeted therapy, exhibited the highest breast lesion SUVmean. Brain and lymph node lesions demonstrated intratumoral heterogeneity of HER2 expression. Qualification of multi parametric maps is anticipated to inform on intratumoral heterogeneity DISCUSSION/SIGNIFICANCE: Despite limitation in clinical applications of quantitative approaches due to lack of standardization of processing, initial investigations, in combining molecular imaging of HER2 and quantitative MRI demonstrate potential in characterizing metastatic HER2+ breast cancer for intratumoral classification and therapeutic stratification.

OBJECTIVES/SPECIFIC AIMS: As pediatric care becomes more concentrated in large urban hospitals, smaller rural hospitals with reduced pediatric care capacity may opt to transfer pediatric patients to higher levels of care even if the patient has a condition that is manageable in a general ED. Up to 20-40% of pediatric transfers are considered avoidable, placing a burden on the patient, their family and the health care system. The aim of this study is to determine the association between pediatric readiness (as measured by the National Pediatric Readiness Project score) and risk of interfacility transfer. We hypothesized that emergency departments with higher measures of pediatric readiness would be less likely to transfer pediatric patients to another facility. METHODS/STUDY POPULATION: The most recent and complete National Pediatric Readiness Project data were from 300 California hospitals in 2012. These data include the overall pediatric readiness score, presence of inter-facility guidelines (written protocols on patients needing care not available at the hospital), presence of interfacility agreements (written agreements with other hospitals regarding transfer patients), and other variables on the facility’s capacity to care for pediatric patients. We linked these hospital data with patient encounter data from the California Office of Statewide Health Planning and Development Emergency Department (ED) Database (OSHPD) using hospital name and zip code. To be eligible for the study, the patient must have a documented ED encounter and be less than 18 years old during the encounter. A patient was considered a transfer patient if they were transferred to a separate general hospital, children’s hospital, federal health care facility, rehabilitation facility, Critical Access Hospital or psychiatric hospital. Chi-square tests and t-tests were used for descriptive statistics. For non-normal data, we used the non-parametric Kruskall-Wallis test. We also used logistic regression to compare the odds of transfer between comparison groups. Statistical analyses were conducted in the R environment version 3.4.4. RESULTS/ANTICIPATED RESULTS: In 2012, there were 2,604,723 pediatric ED encounters, of which 10,966 resulted in a transfer (0.4%). Transferred patients on average were older (15 vs. 5 years, p < 0.001) and more likely to be female (58.6% vs 46.6%, p < 0.001). The transfer group originated from hospitals with a lower median pediatric readiness score (76.0 vs 78.3, p < 0.001). Patients were less likely to be transferred if they were seen at a hospital with written guidelines with transfer protocols (OR 0.89, 95% CI 0.83—0.95, p < 0.001). Patients were more likely to be transferred if they were seen at a hospital with written interfacility agreements with other hospitals (OR 1.17, 95%CI 1.10—1.25, p < 0.001). We anticipate, that even with more sophisticated multilevel statistical models, pediatric readiness scores will remain associated with odds of transfer. DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary data suggest that hospitals with higher levels of pediatric readiness and written guidelines with transfer protocols are less likely to transfer pediatric patients. There may be actionable policy and procedural items that a hospital could enact to lower the rate of transfer patients. Future analyses will include more complex statistical modeling to adjust for confounders, will include inpatient data, and will compare the risk of potentially avoidable transfers between hospitals with varying levels of pediatric readiness.

OBJECTIVES/GOALS: To determine the relationship among serum concentration of tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) and mortality in community-acquired pneumonia (CAP) patients. METHODS/STUDY POPULATION: This is a multicenter 2-year cohort study in Spain, designed to better understand the role of sTWEAK concentrations in CAP patients. CAP patients were prospectively enrolled in two University hospitals and sTWEAK was measured within the first 24 hours of ICU admission. Samples were collected and stored for laboratory analyses. To detect sTWEAK in human samples, we used a commercially available ELISA kit following manufacturer’s instructions. Demographic patients’ characteristics and ICU mortality were prospectively collected. Descriptive statistics and logistical regressions were used to assess the proposed aims. RESULTS/ANTICIPATED RESULTS: A total of forty-three patients were included in the study (10 healthy users, 10 uninfected controls and 23 CAP patients). In comparison to healthy volunteers, patients admitted to the hospital (both, infected and non-infected) had lower level of sTWEAK. During hospital admission, 7 (17%) patients died. Patients whom died during ICU stay due to CAP, had significantly lower levels of sTWEAK when comparing with patients whom survived (Median [IQR]; 509.35 [357.49, 953.92] Vs 1103.03 [716.93, 1663.16]; p = 0.015). In contrast, patients that developed shock did not have different concentrations of sTWEAK (Median [IQR]; 1008.04 [531.87, 1390.80] Vs 1062.29 [575.24, 1598.83], p = 0.84). DISCUSSION/SIGNIFICANCE OF IMPACT: Community-acquired pneumonia (CAP) is the first cause of death in underdeveloped countries. CAP is a pulmonary infection that creates a proinflammatory environment not just locally but also systemically, secondary to upregulation of molecular cascades with a wide variety of proteins being released perpetuating this inflammation and tissue damage. Several of these molecules have been described and linked to a greater risk of inhospital complications, longer length of hospital stay and mortality. TNF-like weak inducer of apoptosis (TWEAK) is a member of the TNF-alpha superfamily, involved in immune response, cell growth, angiogenesis, NF-kB activation and apoptosis induction in tumor cells. It is known that serum-TWEAK plays a role in inflammatory processes, however, its behavior is unknown in patients with CAP. Therefore, this study aims to identify whether there is a relationship between serum concentration of TWEAK and prognosis in CAP patients. To our knowledge, this is the first study to shown that concentration of sTWEAK within the first 24 hours of ICU admission is lower in patients with CAP. Moreover, patients whom died during ICU admission due to CAP, have lower sTWEAK levels. This biomarker may identify patients at higher risk of dying due to CAP and may represent severe CAP. However, further studies are needed to confirm these findings.

OBJECTIVES/SPECIFIC AIMS: The goal of this study was to investigate the associations of systolic blood pressure (SBP), diastolic blood pressure (DBP) and pulse pressure (PP) with knee articular cartilage composition using magnetic resonance imaging (MRI)-based T2 relaxation time measurements in study participants from the Osteoarthritis Initiative (OAI). METHODS/STUDY POPULATION: In this longitudinal study, 1,139 participants from the OAI, a multi-center, observational study of the evolution of knee OA, were selected using the following inclusion criteria: right knee Kellgren Lawrence (KL) score (radiographic classification of OA severity) 0-2 indicating no to mild radiographic OA at baseline, no history of rheumatoid arthritis at baseline, available blood pressure measurements at baseline, available T2 measurements in at least three knee compartments at baseline and 48-month follow-up. Linear regression models were performed using standardized values for SBP, DBP and PP as primary predictors and change in cartilage T2 over 48 months, a measure of cartilage matrix quality and degeneration, as the primary outcome. PP was defined as SBP minus DBP. Change in superficial layer and deep layer cartilage T2, which reflect differences in the laminar organization of knee cartilage T2, were also included as outcomes. Statistical models were adjusted for common risk factors for knee OA (baseline age, sex, BMI, KL score) as well as number of currently used anti-hypertensive medications (AHM) reported at baseline. We included AHMs whose primary indication was the treatment of hypertension including beta blockers, angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARB), thiazides, chlorthalidone, dihydropyridine calcium channel blockers (CCB) and aliskiren. All predictors, outcomes and covariates (except sex) were analyzed as continuous variables. We included interaction terms in the models to evaluate whether the covariates (age, sex, BMI, KL score, number of AHMs) modified the association of SBP, DBP and PP with cartilage T2. RESULTS/ANTICIPATED RESULTS: The average age of all study participants was 58.8 years (SD ± 8.6) with a higher proportion of men (59.4%), average body mass index (BMI) was 28.3 (SD ± 4.5), average SBP was 122.4 (SD ± 15.4) mmHg, average DBP was 75.5 (SD ± 9.6) mmHg and 469 (38.1%) study participants were taking at least one AHM. Higher baseline DBP was significantly associated with a faster increase in global T2 (0.22 [0.10,0.35], P < 0.001), global deep layer T2 (0.20 [0.03,0.36], P < 0.022) and global superficial layer T2 (0.39 [0.20,0.58], P < 0.001). These associations were significant in both unadjusted and the models adjusted for age, sex, BMI and KL score. No significant associations were found between SBP or PP and cartilage T2 and no significant interactions were found between SBP, DBP, PP and the covariates. DISCUSSION/SIGNIFICANCE OF IMPACT: Higher baseline DBP was associated with a faster increase in knee cartilage T2, suggesting accelerated cartilage degeneration. This association was stronger for the superficial layer of knee cartilage T2 compared to the deep layer. Although further basic mechanistic studies are needed to elucidate the underlying pathophysiology of this relationship, these results suggest lowering DBP may influence knee OA.

OBJECTIVES/GOALS: Buprenorphine (BUP) is used for opioid use disorder during pregnancy but causes neonatal opioid withdrawal syndrome (NOWS). The goal of this study was to determine the contribution of the active metabolite, norbuprenorphine (NorBUP), to the development of NOWS when the parent drug, BUP, is administered during pregnancy. METHODS/STUDY POPULATION: Subcutaneously implanted osmotic minipumps delivered BUP (0, 0.01, 0.1 or 1 mg/kg/day) ± NorBUP (1 mg/kg/day) to pregnant Long-Evans rats from gestation day 9 until after delivery. NOWS was measured between 3 and 12 hours after delivery. Withdrawal was precipitated by an intraperitoneal injection of a mu opioid receptor antagonist naltrexone (NTX; 0, 1 or 10 mg/kg), and movement duration (MD; a validated proxy for NOWS) was measured using Noldus Ethovision. Concentrations of BUP, NorBUP, and their glucuronide conjugates in the brains of neonatal littermates not undergoing withdrawal testing were determined using LC/MS/MS. Two-way ANOVA and multiple linear regression analyses tested for interactions between BUP and NorBUP on MD and related brain concentrations to MD, respectively. RESULTS/ANTICIPATED RESULTS: There was no interaction effect between BUP and NorBUP on MD for either sex or at any dose of naltrexone. In females, but not males, BUP (1 mg/kg/day) significantly increased NorBUP-induced MD by 58% following an injection with 1 mg/kg NTX. A multiple linear regression model that included BUP and NorBUP brain concentrations as predictors of MD was significant and well-fitting [FEMALES: F (2, 40) = 23.97, P < .0001, adj R2 = 0.52; MALES: F (2, 40) = 5.84, P = .0059, adj R2 = 0.19]. There was a differential contribution of NorBUP brain concentrations to MD based on sex. The partial regression coefficient for NorBUP was 51.34 (p < .0001) for females and 19.21 (p = 0.093) for males. The partial regression coefficient for BUP was similar for females and males (FEMALES:βBUP = 10.62, p = .0017; MALES:βBUP = 11.38, p = .009). DISCUSSION/SIGNIFICANCE: We show for the first time a differential contribution of NorBUP to BUP-associated NOWS in each sex, suggesting sex differences in NorBUP susceptibility and implicating that treatment strategies reducing prenatal NorBUP exposure may be more effective for females than males.

ABSTRACT IMPACT: This study will help determine whether ecological momentary assessment is feasible in assessing changes in negative affect and the occurrence of non-suicidal self-injury (NSSI) in military Veterans with post-traumatic stress disorder; if so it will allow for further examination of correlates of NSSI which will inform treatment efforts. OBJECTIVES/GOALS: Given the advantages of ecological momentary assessment (EMA), and the lack of research on non-suicidal self-injury (NSSI) in military populations, the goal of the current pilot study is to determine the feasibility of using EMA to assess daily changes in post-traumatic stress disorder (PTSD) symptoms, negative affect and NSSI in veterans with PTSD. METHODS/STUDY POPULATION: Twenty military veterans with post-traumatic stress disorder (PTSD) who have engaged in non-suicidal self-injury (NSSI) in the previous 12 months will be recruited. Participants will complete assessments 4 times per day for 28 days at randomly scheduled times. Assessments will measure PTSD symptoms, negative emotions, and NSSI urges and behaviors. At the conclusion of the 28-day study period, participants will complete measures that ask about their experiences in the study, (e.g., the acceptability of the daily surveys and the accessibility of the web-based surveys). Feasibility will be determined with regard to the success of recruiting eligible participants and compliance with daily survey completion. Variability in PTSD symptoms, negative affect, and NSSI urges and behaviors also will be determined. RESULTS/ANTICIPATED RESULTS: It is anticipated that this study will successfully recruit 20 veterans with PTSD with a history of engaging in NSSI within the previous 12 months. It is also anticipated that daily survey completion rates will be approximately 90% based on previous research using EMA with veterans with PTSD and that participants will indicate satisfaction with the procedures of the study. It is anticipated that participants will demonstrate variability in PTSD symptoms, indicated by changes in the number of symptoms endorsed and the intensity of those symptoms experienced. Finally, it is anticipated that participants will demonstrate variability in negative emotions (fear, hostility, guilt, and sadness).

DISCUSSION/SIGNIFICANCE OF FINDINGS: Findings from this study will support the use of EMA in a subsequent large-scale investigation examining time-varying symptoms of PTSD and negative affect as antecedents to NSSI. Information from this large-scale study will in turn be used to inform treatments that may to decrease NSSI in veterans by targeting specific symptoms and negative emotions.

OBJECTIVES/GOALS: Asthma is a significant health concern that affects people of all ages worldwide. EAS demonstrates many of the pathophysiological characteristics of nonatopic human asthma, which has led EAS to be used as naturally occurring model. Previous work from our lab determined that MARCKS (Myristoylated Alanine Rich C Kinase Substrate) protein is an essential regulator of cellular inflammatory functions. In the current study, we hypothesized that MARCKS levels would be increased in BAL cell lysates from horses with EAS, and that inhibition of MARCKS in zymosan-stimulated BAL cells (ex vivo) would diminish respiratory burst. METHODS/STUDY POPULATION: Lysates were prepared from BAL cells isolated from horses with no, mild/moderate and severe EAS. Relative MARCKS protein levels were determined using equine specific MARCKS ELISA (MyBioSource). Cultured BAL cells were pretreated with a MARCKS inhibitor peptide (MANS), control peptide (RNS) or vehicle control and stimulated with zymosan for 5 hours. Reactive oxygen species levels were determined by luminescence to evaluate respiratory burst. Data were analyzed by One-way ANOVA (p<0.05). RESULTS/ANTICIPATED RESULTS: We determined that normalized MARCKS protein expression is significantly increased in BAL cell lysates from horses with mild/moderate or severe EAS, compared to horses with normal BAL cytology. Preliminary findings also suggest that MANS treatment of zymosan-stimulated equine BAL cells ex vivo attenuates levels respiratory burst. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings point to a possible role for MARCKS protein in the pathophysiology of EAS and support MARCKS inhibition as a potential therapeutic strategy.

OBJECTIVES/GOALS: The cytoskeletal protein α²II spectrin interacts with actin and ankyrin-2 in cardiomyocytes which is essential to orchestrate ion channels and membrane proteins in the cardiac dyad. Our goal is to understand molecular mechanism causing severe ventricular arrhythmias due to spectrin dysfunction and explore novel therapies to treat such conditions. METHODS/STUDY POPULATION: We previously published a case of a 36-year-old woman with an ankyrin-2 p.R990Q (ANK2) variant, presented with severe ventricular arrhythmias and sudden cardiac arrest, caused by a novel mutation in the ankyrin-B gene (c.2969G>A) that disrupts the interaction of ankyrin-B/βII spectrin. To model the condition, we will use human induced pluripotent stem cell (DF 19-9-7T, WiCell)-derived ventricular cardiomyocytes (iPSC-CMs) having ANK2 variant, engineered using CRISPR/Cas9 method (Synthego Corp.). We will validate the differentiation of iPSCs into ventricular lineage and characterize the ANK2 ventricular phenotype. Next, we will express light-gated cation channel Channelrhodopsin (ChR2) in the ANK2 iPSC-CMs and investigate the potential role of optogenetics in treating such severe arrhythmias. RESULTS/ANTICIPATED RESULTS: Immunostaining shows 87.339% of iPSC-CMs, treated with All-trans retinoic acid (RA) (1 uM) on days 7 and 12 [RA 7,12], and 23.84% of those, treated on days 3 and 5 [RA 3,5], expressed MLC-2V (p<0.001). Calcium reuptake (τ) is 0.5914 s in RA 7, 12 while 0.2247s in RA 3, 5 (p<0.001). APD90 and APD50 of RA 7, 12 are 2- and 5-fold higher than RA 3, 5, showing distinct ventricular and atrial phenotypes. Protein expression of βII-spectrin and ankyrin-2 and their co-localizations were reduced in the ANK2 phenotype compared to the healthy phenotype. We found prolongation of Ca2+ waves and τ with blue light on iPSC-CMs, expressing ChR2. We anticipate that such prolongation of calcium transients would prevent aberrant calcium spikes, rescue Ca2+/calpain-induced βII-spectrin loss and provide electrical stability. DISCUSSION/SIGNIFICANCE: Animal models cannot accurately recapitulate human cardiac electrophysiology. The proposed human iPSC-CM-based ANK2 model would provide better mechanistic insights of severe ventricular arrhythmias. Also, the proposed optogenetic cardioversion has the potential to provide safe, targeted and painless cardioversion to manage arrhythmias.

Given the dramatic growth in the financial burden of cancer care over the past decades, individuals with cancer are increasingly susceptible to developing social needs (e.g., housing instability and food insecurity) and experiencing an adverse impact of these needs on care management and health outcomes. However, resources required to connect individuals with needed social and community services typically exceed the available staffing within clinical teams. Using input from focus groups, key informant interviews, user experience/user interface testing, and a multidisciplinary community advisory board, we developed a new technology solution, ConnectedNest, which connects individuals in need to community based organizations (CBOs) that provide services through direct and/or oncology team referrals, with interfaces to support all three groups (patients, CBOs, and oncology care teams). After prototype development, we conducted usability testing, with participants noting the importance of the technology for filling a current gap in screening and connecting individuals with cancer with needed social and community services. We employ a patient-empowered approach that engages the support of an individual’s healthcare team and community organizations. Future work will examine the integration and implementation of ConnectedNest for oncology patients, oncology care teams, and cancer-focused CBOs to build capacity for effectively addressing distress in this population.

Large translational research initiatives can strengthen efficiencies and support science with enhanced impact when practical conceptual models guide their design, implementation, and evaluation. The National Institutes of Health (NIH) Environmental influences on Child Health Outcomes (ECHO) program brings together data from 72 ongoing maternal–child cohort studies – involving more than 50,000 children and over 1200 investigators – to conduct transdisciplinary solution-oriented research that addresses how early environmental exposures influence child health. ECHO uses a multi-team system approach to consortium-wide data collection and analysis to generate original research that informs programs, policies, and practices to enhance children’s health. Here, we share two conceptual models informed by ECHO’s experiences and the Science of Team Science. The first conceptual model illuminates a system of teams and associated tasks that support collaboration toward shared scientific goals. The second conceptual model provides a framework for designing evaluations for continuous quality improvement of manuscript writing teams. Together, the two conceptual models offer guidance for the design, implementation, and evaluation of translational and transdisciplinary multi-team research initiatives.

OBJECTIVES/GOALS: The goal of this project is to identify new therapeutic targets and biomarkers to treat or prevent cocaine toxicity by investigating proteomic, transcriptomic and epigenetic signatures of cocaine exposure in human subjects. METHODS/STUDY POPULATION: Cocaine is a highly addictive neurotoxic substance, and it is estimated that 1.9 million Americans are current users of cocaine. To study the molecular effects of cocaine, we generated preliminary proteomics and next-generation RNA sequencing (RNAseq) data from human postmortem dorsolateral prefrontal cortex (Broadmann area 9 or BA9) of 12 cocaine-exposed subjects and 17 controls. Future directions for this project include RNAseq and DNA methylation analysis of neuronal nuclei sorted from human postmortem BA9 and a human induced pluripotent stem cell-derived neuron (hiPSN) model of cocaine exposure from the same postmortem subjects from whom we have brain samples. RESULTS/ANTICIPATED RESULTS: We found alterations in neuronal synaptic protein levels and gene expression, including the serotonin transporter SLC6A4, and synaptic proteins SNAP25, SYN2, SYNGR3. Pathway analysis of our results revealed alterations in specific pathways involved with neuronal function including voltage-gated calcium channels, and GABA receptor signaling. In the future, we expect to see an enhancement in neuron-specific gene expression signatures in our sorted neuronal nuclei and our hiPSN model of cocaine exposure. The hiPSN model will help elucidate which effects are due to acute versus chronic exposure of cocaine. DISCUSSION/SIGNIFICANCE OF IMPACT: Neuronal signatures found with this analysis can help us understand mechanisms of cognitive decline in long-term cocaine users as well as the acute effects on the brain of cocaine taken in overdose. With this work and future proposed studies, we can discover novel clinical biomarkers for cocaine neurotoxicity in patients with cocaine use disorder and determine readouts for future therapeutic development on cocaine addiction and overdose.

ABSTRACT IMPACT: We present a minimally-invasive approach to monitoring heavy metal exposures in geographically disperse populations; this framework may encourage and facilitate greater participation in a broad range of related clinical studies that require biosampling. OBJECTIVES/GOALS: We hypothesize that microneedle array (MA) extraction of interstitial fluid (ISF) will enable minimally-invasive quantitation of heavy metal (HM) exposure. We establish analytical parameters for ICP-MS analysis of HMs in ISF, quantify baseline HM content in ISF vs other fluids, and characterize a mixed HM exposure model. METHODS/STUDY POPULATION: Recent advances in ISF extraction and analysis suggest a minimally-invasive to monitor HM exposure longitudinally in both urban and dispersed communities. ISF can be collected with MAs and is a rich source of disease and exposure biomarkers. To refine analytical methods, Human subjects with no underlying skin disease were recruited into the IRB-approved study. Each subject had blood and urine collected. ISF was also collected using 3D-printed MA-holders. The fluids were analyzed, using ICP-MS, to quantify the levels of uranium (U), cadmium (Cd), vanadium (V), and arsenic (As). Additionally, we analyzed 2,770 subjects from the public NHANES dataset from 2018-2019. Python and Scikit-learn were used to analyze the demographics, survey responses, and metal concentrations for these individuals. RESULTS/ANTICIPATED RESULTS: While several studies have described the surface water and sediment content of toxic metals, determining biological loads remains challenging due to the need to collect blood or urine from a dispersed rural population over time. Our preliminary results suggest similar HM concentrations in ISF, compared with blood in a small unexposed population. Analysis of subjects from the NHANES public datasets suggest similar baseline blood HM concentrations in diverse subject populations with some differences in Cd depending on smoking and e-cigarette usage. Correlation maps also suggest possible synergy between different metals with cobalt and chromium showing the highest correlation. The initial results from this study have been applied to develop a mixed HM exposure model in rats for further translational testing. DISCUSSION/SIGNIFICANCE OF FINDINGS: We present a minimally-invasive HM monitoring approach. Exposure to multiple HMs is suspected to have additive or synergistic harmful health effects. We ultimately envision a wearable microneedle patch that could be mailed to individuals or distributed through community centers, worn for a few hours, and returned to a central laboratory.

OBJECTIVES/GOALS: 30,000,000 people in the U.S. have hearing loss, negatively impacting quality of life and work. Understanding auditory axon guidance for spiral ganglia neurons (SGNs) will aid development of new therapies. I study role of Eph/Ephrin signaling in mediating type II SGN turning events, and how planar cell polarity (PCP) signaling modulates this process. METHODS/STUDY POPULATION: This quantitative study was conducted on Efna3 and Vangl2 null mice possessing Neurog1CreERT2 and R26RtdTomato mutations. Spontaneous Cre activity within the Neurogenin 1 CreERT2 line causes recombination and expression of fluorescent Rosa26 Reporter (R26R)tdTomato in a restricted number of SGNs, including type IIs. Together these lines permit SGN sparse labeling. Bulk-labeling was used for Efna3;Vangl2 double knockout (DKO) mutants. Immunostaining and confocal imaging was used to analyze dsRed in Efna3; Vangl2 and NF-200 in DKOs to quantify type II SGN turning. In combination, 3D rendering in Imaris software was used to quantify type II SGN turning, branching and other growth and navigation characteristics. 5-6 cochleae per genotype were analyzed and compared by t-test to wildtype controls. RESULTS/ANTICIPATED RESULTS: EPHRIN-A3 is expressed on the membranes of outer pillar and Deiters’cells of the cochlear epithelium. Efna3 nulls showed a small rise in type II SGNs incorrectly turning toward the apex at an error frequency of 16.9% compared to controls (n=6; p=0.05). Efna3 nulls had reduced branch number/fiber compared to controls, 4.14 and 7.22, respectively (n=129; p DISCUSSION/SIGNIFICANCE: Our results suggest that Eph/Ephrin signaling acts parallel of PCP signaling to mediate type II SGN guidance during development. The clinical implications of these findings are that therapeutics targeting EPHRIN-A3 and/or VANGL2 in this pathway could stimulate new outer hair cell innervation by type II SGNs following auditory damage.

OBJECTIVES/SPECIFIC AIMS: Promote knowledge translation and evidence-informed decision-making by assessing barriers and facilitators to balancing cost and quality of care within the US state of Maryland and nation of Denmark. METHODS/STUDY POPULATION: Open-ended and semi-structured key-informant interviews were conducted in 2016 and 2017 among high level decision-makers in Maryland (N=21) and the Danish (N=17) healthcare systems, including hospital, local, regional, and cross-organizational administrators and elected officials. The interviews consisted of questions related to: (1) currently practiced and preferred approaches to resource allocation and development and use of quality performance measures, and (2) preferred sources, formats/styles, modes of information, and decision-making strategies based on a shift from volume to quality-driven care. RESULTS/ANTICIPATED RESULTS: Decision-makers in Maryland expressed the need for collaboration in a changing environment, yet increasingly rely on cost and quality outcomes data to drive decisions and note the struggle to identify credible and useful information. Maryland decision-makers also face challenges in regulating utilization and costs without mandated participation of physician practices within the global budget cap model, which is perceived to be a primary driver of healthcare utilization in the hospital sector. Similarly, decision-makers in Denmark conveyed the importance of quantitative data to aid decisions, however, stress collaboration and dialogue as driving factors and important sources of information. Danish decision-makers also express challenges to wide-spread adoption of a quality-driven approach due to unsustained quality assurance regulatory bodies. DISCUSSION/SIGNIFICANCE OF IMPACT: The findings suggest implementation of value-based healthcare is highly driven and influenced by availability of credible data, which may significantly impact development of policies and innovative cost control strategies, and regulatory oversight to promote adoption of quality measures in decision-making. Furthermore, collaboration within and across healthcare organizations remains a key component to health system improvement as it fosters dialogue and sharing of best practices among stakeholders.