OBJECTIVES/GOALS: Compare metabolic health of type 2 diabetics on GLP-1 to those on traditional therapy METHODS/STUDY POPULATION: Outcomes of interest of this study include analyzing GLP-1 agonists on overall metabolic health, focusing primarily on weight loss and ability to wane off insulin without rebounding metabolic health. The data will be collected in a retrospective chart review from medical records of type II diabetics from Children’s of Alabama and will follow patients over two years. The charts have been narrowed to those patients prescribed GLP-1 agonists who have been on the medication for at least one year with consistent visits to the endocrinology clinic.The following data will be collected from the charts:race/ethnicity, date of visit, BMI/weight, A1C, insulin therapy, lipids, LFTs (AST/ALT), and insurance coverage RESULTS/ANTICIPATED RESULTS: With these results, we hope to study the metabolic health effects of GLP-1 agonists on type II diabetes in the pediatric population. It is known that obesity is a risk factor for type II diabetes, and that GLP-1 agonists aid in weight-loss in adults. Further research is needed to see the real world health effects, and with these results we hope to assess if GLP-1 agonist have an affect on metabolic health within the pediatric population by collecting data on values aforementioned. We also hope to compare and contrast the different GLP-1 agonists being used based on adherence, insurance coverage, adverse effects, and patient preference. Currently, only Liraglutide and Exetanide are approved for pediatric type II diabetics. DISCUSSION/SIGNIFICANCE: Insulin use can lead to weight gain; metformin does not aidin weight reductions. If GLP-1 agonists aid in weight loss, it could potentially help slow complications of diabetes in the pediatric population. B-cell function in children declines more rapidly; and, as a result, insulin resistance occurs more rapidly.

OBJECTIVES/GOALS: This study will collect multimodal and longitudinal data in adults with obsessive-compulsive disorder and healthy controls. A mixed effects random forest machine learning approach will be taken to develop a model that can predict individualized longitudinal OCD symptom burden. METHODS/STUDY POPULATION: Baseline resting state functional MRI (rsfMRI) and measures of symptom burden will be collected in adults with OCD and healthy controls. Longitudinal measures of behavior and physiology–such as heart rate, activity, and sleep metrics - will be collected using Fitbit Charge 5 tracker. Daily assessments of symptom burden and functional status will be collected through a smartphone app. Individuals with OCD will start pharmacotherapy during the study period and all participants will be followed for a total of 10 weeks. Repeat rsfMRI imaging will occur at study conclusion. Data will be analyzed using a mixed effects random forest machine learning algorithm with assessment of model performance. RESULTS/ANTICIPATED RESULTS: Prior studies of symptom severity in psychiatric illness and affect in non-clinical populations have found longitudinal features - such as lexical and acoustic measures, participant context, heart rate, and sleep metrics–that were predictive of these states over time. It is anticipated that the present study will extend these results to individuals with OCD and identify physiologic and behavioral features that track personalized symptom burden longitudinally in this patient population. A model able to predict when symptoms are elevated could allow for provision of additional treatment or interventions targeted to times of high symptom burden. DISCUSSION/SIGNIFICANCE: This study will be the first to collect and analyze longitudinal measures of behavior, symptoms, and physiology in patients with OCD with a goal of predicting symptom burden. Identification of elevated symptom burden would allow for implementation of just-in-time treatment, during these periods.

OBJECTIVES/GOALS: Renal vascular resistance (RVR) is the opposition to blood flow by renal arteries. At the population level, dietary salt increases RVR and blood pressure (BP), which are associated with cardiovascular disease. Recent data indicate exogenous ketones may offset adverse cardiorenal effects of salt. METHODS/STUDY POPULATION: Our registered clinical trial (NCT05545501) is a double-blinded, placebo-controlled, crossover study. Participants are being randomized to three 10-day conditions: A) control; B) high salt; C) high salt and ketone supplementation. Ten participants are enrolled (target 30 participants). Renal blood velocity (RBV) in the renal and segmental arteries will be measured in the decubitus position using Doppler ultrasound during a 3-minute baseline and 3-minute cold pressor test. We will measure brachial BP with an automated oscillometric BP monitor. RVR will be calculated as mean BP divided by RBV. Statistical analyses will include ANOVA and correlations with α set at ≤ 0.05. RESULTS/ANTICIPATED RESULTS: We anticipate attenuated RBV and increased BP during the cold pressor test, particularly following high salt loading, leading to greater RVR. We hypothesize ketone supplementation will attenuate the high salt induced increase in RVR during the cold pressor test. In addition to RVR we will examine renal vascular conductance which is the ease with which blood flows through arteries, calculated as RBV divided by mean BP. Additional hemodynamics such as heart rate and systolic and diastolic BP will be reported and correlated with primary outcomes. DISCUSSION/SIGNIFICANCE: Dietary salt plays a role in hypertension, cardiovascular disease, and chronic kidney disease, which are leading causes of death. Ketone supplementation may be a promising approach to counteract the detrimental effects of high dietary salt and improve cardiovascular health in adults.

OBJECTIVES/GOALS: The LHS concept has been promoted as a means for health systems to improve quality, safety, efficiency and equity. NAMs definition has been widely adopted, but is broad and has led to variation in how LHS is operationalized. Drawing on a taxonomy developed through a review of literature, we developed a tool that shows how LHSs are implemented in practice. METHODS/STUDY POPULATION: The LHS Implementation Assessment Tool (LHS-IAT) will indicate which forms of work are being carried out by a health system that purports to operate a LHS. LHS-IAT is based on the LHS Consolidated Framework (LHS-CF); which was developed through a qualitative analysis of LHS literature. LHS-CF contains 38 primary elements’ and 56 secondary elements’ that have been associated with the LHS construct. These elements are organized into 5 bodies of work• (e.g.; translating evidence into practice) and 4 enabling conditions• (e.g.; supportive culture). LHS-IAT assesses whether a health system operating as an LHS is implementing each of the key elements in LHS-CF. The usefulness of LHS-IAT will be demonstrated by applying the tool to 5 LHSs that have been described in the literature. RESULTS/ANTICIPATED RESULTS: LHS-IAT produces a quantitative profile for any given health system operating as a LHS; each LHS element is assessed as either emphasized; otherwise present; or absent. With this information, we create profiles for each implementation of LHS, using spider graphs. Systems that emphasize different elements will have different shapes for their spider graphs. Based on our initial coding of publications, we expect at least 4 distinct profiles within our sample, reflecting differences in emphasis on factors such as: continuous improvement practices, adoption of internally and externally tested interventions, research conducted to address patient care issues prioritized by institutional leaders, investigator-initiated research, clinician-engaged research, and engagement of patients and families. DISCUSSION/SIGNIFICANCE: The LHS-IAT will show differences in how health systems are translating the LHS concept into practice. This will allow for a shared language for those studying and/or implementing LHS. With the ability to map out an approach, health system leaders will have a tool to clarify intent and gain consensus as to which LHS model they want to implement and invest in.

ABSTRACT IMPACT: It provides insight in the relationship between pediatrics and clinical research and how pediatric participation in CT translates to clinical significance in form of drug labels, which inform clinicians on how to prescribe pediatric medications. OBJECTIVES/GOALS: Assessing the extent that the Best Pharmaceuticals for Children Act (BPCA) advances pediatric inclusion in clinical trials (CTs) and the availability of pediatric-specific drug information METHODS/STUDY POPULATION: The BPCA provides the U.S. Food and Drug Administration (FDA) authority to solicit sponsors whose drugs may benefit pediatric populations. Participation is voluntary and provides additional market exclusivity and pediatric information. CTs that received marketing exclusivity from 2016-2018 under BPCA were reviewed using Clinicaltrials.gov to access the legislation’s impact. CTs were categorized according to eligibility: (1) pediatric and adult groups, (2) pediatrics, and (3) pediatric sub-groups. Studies were excluded for ambiguous age data. Studies open to both groups were evaluated for pediatric participation. Each drug was searched in DailyMed.com for published pediatric indications. RESULTS/ANTICIPATED RESULTS: Between 2016 - 2018, 22 drugs received marketing exclusivity under BPCA. Of the 196 CTs conducted for these drugs, 135 were available to adults and pediatrics, 10 were available to the entire pediatric population, and 51 were available to specific pediatric sub-populations. Exclusion criteria permitted only 118 of the CTs for assessment where eligibility included both pediatric and adult populations, of which 65 of these had less than 1% pediatric representation. Of the 22 drugs, 20 have pediatric indications. Over this three-year period, the number of CTs where adults and pediatrics were eligible were greater than CTs for pediatric only or pediatric subpopulations. DISCUSSION/SIGNIFICANCE OF FINDINGS: It is prevalent for BPCA compliant CTs to include both; 65% of drugs (13/20) with pediatric indications had more studies involving both groups than only pediatrics. Adequate pediatric CT representation is necessary for developing pediatric drug labeling with meaningful data for clinical indications.

OBJECTIVES/GOALS: Clinical tissue specimens are primarily destined for formalin fixed, paraffin embedded processing to create a basis for diagnosis by microscopic examination. Innovations in specimen processing are required to expand its availability for inclusion as the substrate in assays that can contribute to the further development of Precision Medicine. METHODS/STUDY POPULATION: Transurethral resection of bladder tumors were selected for testing based on availability and tissue composition. A wash step was used to generate daughter aliquots composed of dislodged cells and a solution with prior contact to the parent tissue. This wash step served two purposes: 1) reduce the amount of contaminating material from spreading to other cases, a problem known to be associated with this type of specimen; and 2) create aliquots from which additional informative data could be generated. These daughter aliquots were then examined to determine their value as a source for exosome profiling, metabolomic studies, molecular characterization and organoid development. The parent tissue was not compromised, was able to undergo conventional processing and yielded results equivalent to unwashed specimens. RESULTS/ANTICIPATED RESULTS: Exosomes secreted by the tumor cells were identified to be present in the daughter aliquots by a combination of their isolation using CD31 and detection of miR-21 expression. These exosomes were confirmed to be not related to fragmented cells from testing for beta-tubulin. A global/discovery-based approach using mass spectrometry provided insights into early characterization of metabolomic profiles present in these tumor cells. Ample amounts of high quality DNA (226 ng/ul concentrations; 11.3 ug total) were recovered from the dislodged, excess cells in the wash for molecular studies. Finally, from viable cells recovered in one of the daughter wash aliquots, the ability to grow organoids was proven to be possible and reproducible. DISCUSSION/SIGNIFICANCE: Based on these results, the value of the clinical specimen can be markedly expanded for utilization in research and possible clinical use without detracting from the parent tissue. This non-destructive, easy to adopt wash procedure can potentially lead to an influx of data that may ultimately prove useful in improving patient care.

OBJECTIVES/GOALS: Our goal is to determine if a precision medicine approach to guide anti-platelet therapy for patients with ACS, post PCI, is feasible for a diverse urban population. Also, we will evaluate if guided therapy reduces major adverse cardiovascular events (MACE) while remaining economically sustainable. METHODS/STUDY POPULATION: This prospective, pragmatic study will enroll two-hundred patients with ACS undergoing PCI, receiving DAPT. Patients will receive point-of-care CYP2C19 genotyping. Patients with at least one loss-of-function (LoF) allele will be recommended prasugrel. Those without LoF alleles, will be recommended to take prasugrel for 7 days then clopidogrel for 7 days, followed with platelet reactivity phenotyping. Patients with HPR >208 P2Y12 reaction units will take prasugrel; the remainder will take clopidogrel. We will review electronic health records and contact patients at baseline, then at 1, 3, 6, and 12 months to collect data for cardiovascular and health-related quality of life (HRQoL) outcomes. RESULTS/ANTICIPATED RESULTS: Feasibility and clinical utility will be measured by the proportion of patients with a genotype or phenotype leading to a clinical recommendation of alternative therapy and whether or not recommendations were accepted by clinicians. Effectiveness will be measured by combined MACE (composite of cardiovascular death, non-fatal myocardial infarction, and non-fatal stroke), stent thrombosis, and major and minor bleeding over the study period. Cost of testing, 30-day hospital readmission, and HRQoL questionnaires will be included for pharmacoeconomic analysis from an institutional perspective. DISCUSSION/SIGNIFICANCE OF IMPACT: There are no studies investigating the clinical utility of implementing guided anti-platelet selection, combining CYP2C19 genotyping and HPR phenotyping. We anticipate incorporating this precision medicine approach to guide P2Y12 inhibitor selection will be feasible while improving patient outcomes.

Interprofessional healthcare team function is critical to the effective delivery of patient care. Team members must possess teamwork competencies, as team function impacts patient, staff, team, and healthcare organizational outcomes. There is evidence that team training is beneficial; however, consensus on the optimal training content, methods, and evaluation is lacking. This manuscript will focus on training content. Team science and training research indicates that an effective team training program must be founded upon teamwork competencies. The Team FIRST framework asserts there are 10 teamwork competencies essential for healthcare providers: recognizing criticality of teamwork, creating a psychologically safe environment, structured communication, closed-loop communication, asking clarifying questions, sharing unique information, optimizing team mental models, mutual trust, mutual performance monitoring, and reflection/debriefing. The Team FIRST framework was conceptualized to instill these evidence-based teamwork competencies in healthcare professionals to improve interprofessional collaboration. This framework is founded in validated team science research and serves future efforts to develop and pilot educational strategies that educate healthcare workers on these competencies.

OBJECTIVES/GOALS: 48,000,000 people in the U.S. have hearing loss, negatively impacting quality of life and work. Unveiling axon guidance for auditory type II spiral ganglia neurons (SGNs) will aid development of new therapies. I study the role of Eph/Ephrin and planar cell polarity (PCP) signaling during type II SGN turning and outer hair cell (OHC) innervation. METHODS/STUDY POPULATION: This quantitative study was conducted on Efna3 and Vangl2 null mice possessing Neurog1CreERT2 and R26RtdTomato mutations. Spontaneous Cre activity within the Neurogenin1CreERT2 line causes recombination and expression of fluorescent Rosa26 Reporter (R26R)tdTomatoin a restricted number of SGNs, including type IIs. Together, these lines permit SGN sparse labeling. Immunostaining and confocal imaging were used to analyze dsRed in Efna3 and Vangl2 mice and quantify type II SGN turning. In combination, Imaris 3D renderings were used to quantify type II SGN turning, branching, navigation features and temporal effects of EPHRIN-A3-Fc on type IIs via cochlear cultures (a gain-of-function manipulation). For both sexes, 5-6 cochleae per genotype were analyzed and compared by t-test to wildtype (WT) controls. RESULTS/ANTICIPATED RESULTS: Efna3 nulls showed a small rise in type II SGNs incorrectly turning toward the apex at an error rate of 16.0% compared to WTs (n=6; p=0.05). P0 Efna3 nulls had reduced branch number compared to WTs, 4.1 and 7.2, respectively (n=129; p=<0.0001), suggesting EPHRIN-A3 acts as a positive growth cue. In cochlear cultures, EPHRIN-A3-Fc led to type II SGN collapse at E15.5, indicating a repulsive function. However, at P0, EPHRIN-A3-Fc treatment led to type II SGNs with elevated branch numbers compared to Control-Fc treatment: 18.1 and 11.4, respectively (n=116; p=<0.0001). This indicates a positive growth function. At E16.5, EPHRIN-A3 protein immunoreactivity on Deiters’ and pillar cells appears reduced in Vangl2 nulls compared to WT cochleae, suggesting that EPHRIN-A3 acts downstream of PCP signaling. DISCUSSION/SIGNIFICANCE: Results suggest that Eph/Ephrin signaling acts downstream of PCP signaling to mediate type II SGN guidance and EPHRIN-A3 switches its mode of activation. The clinical implications of these findings are that therapeutics targeting EPHRIN-A3 and/or VANGL2 in their given pathways could stimulate new OHC innervation following auditory damage.

OBJECTIVES/SPECIFIC AIMS: In the present study, we examined the functional consequences of 3α-PVA on OATP1B1-mediated PVA transport. To elucidate this, we determined the effect of SLCO1B1 genotype on PVA transport and the role of 3αPVA as a competitive inhibitor of OATP1B1, which could serve as another covariate that disrupts the systemic and hepatic exposure of pravastatin in children and adults. METHODS/STUDY POPULATION: Site directed mutagenesis was performed to generate SLCO1B1 genotypes of interest (*1a, *1b, *5, *15). Human embryonic kidney (HEK293) cells were grown and plated at 200 000 cells per well in 24-well plates. Twenty-four hours later the cells were transfected with the aforementioned plasmids. Forty-eight hours later cell-based transport was performed with radiolabelled [3H]-pravastatin sodium salt. Non-radioactive pravastatin sodium salt and 3’α-iso-pravastatin sodium salt was used for PVA transport and 3αPVA studies, respectively. Cells were washed 3 times with warm uptake buffer, incubated for 1 minute with uptake solutions containing PVA and 3αPVA at varying concentrations. Transport was terminated by four 1-ml washes with ice-cold uptake buffer. Cells were lysed with 300 µl 1% Triton X-100 in PBS at room temperature for 30 minutes prior to analysis. Radioactivity was measured in a MicroBeta2 liquid scintillation counter. The remaining cell lysates were transferred to 96-well plates to determine total protein concentration using the bicinchonic acid protein assay. All transport measurements were corrected by the total protein concentration. All experiments were performed 3 to 4 times independently with 2-3 determinations. Data were analyzed for significant differences amongst genotype groups using ANOVA followed by Tukey’s multiple comparisons test. IC50 and kinetic parameters were calculated using non-linear regression analysis. RESULTS/ANTICIPATED RESULTS: Pravastatin transport in SLCO1B1 variants (*5, *15) was significantly decreased compared to the reference genotype *1a and *1b (Km [µM]: *1a 18.2 ± 0.9; *1b 17.9 ± 3.3; *5 34.2 ± 9.7; *15 34.1 ± 6.1; p≤0.05; Vmax [pmol/mg/min]: *1a 104.9 ± 13.1; *1b 93.7 ± 16.7; *5 44.8 ± 15.9; *15 62.3 ± 22.5; p≤0.05). *1a and *1b were not significantly different with respect to pravastatin transport. Intrinsic clearance was diminished nearly 4 to 5-fold in SLCO1B1 variants compared to reference genotypes (Vmax/Km [µl/min/mg]: *1a 5.8 ± 0.8; *1b 5.7 ± 1.9; *5 1.3 ± 0.2; *15 1.8 ± 0.3; p≤0.01). Pravastatin transport was inhibited by 3αPVA for all genotypes. However, there was more pronounced inhibition in the SLCO1B1 variant genotypes compared to reference genotypes (IC50 [µM]: *1a 15.9 ± 1.9; *1b 18.6 ± 5.7; *5 3.9 ± 2.0; *15 4.4 ± 0.8; p≤0.01; Ki: *1a 15.0 ± 1.8; *1b 17.5 ± 5.4; *5 3.8 ± 2.9; *15 4.3 ± 0.8; p≤0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: In vitro PVA transport is altered according to SLCO1B1 genotype, consistent with previous in vitro and human experience. Our data suggest that the significantly different maximal transport velocity (Vmax) in variant versus non-variant genotypes is consistent with decreased membrane expression of OATP1B1 with the variant c.521T>C allele. However, in contrast to data involving typical model substrates (e.g. estrone-3-sulfate), the PVA binding affinity (Km) was significantly different between variant and non-variant genotypes, consistent with altered binding of the substrate to OATP1B1. Collectively, we conclude that decreased OATP1B1 expression and function in variant genotypes influence altered transport for PVA. Finally, the functional consequences of 3αPVA formation on PVA transport was confirmed in our study. Mechanistically, we confirmed our observation in humans that 3αPVA inhibits OATP1B1 transport. However, this effect is more pronounced in variant genotypes as shown by lower IC50 values compared to the reference genotypes. This highlights another source of variation that must be taken into consideration when trying to optimize the pravastatin dose-exposure relationship in humans.

OBJECTIVES/GOALS: CTRUs support clinical research. RUN is a Learning Research System that is created to enhance CTSA and non-CTSA research units capacity through implementing, assessing, and disseminating discoveries in methods, approaches, education, and training in clinical and translational science. METHODS/STUDY POPULATION: The RUN association began in July 2018 with eight universities. The association has grown to 44 hospitals, research, and academic institutions (including 36 CTSA institutions). A RUN Discussion Forum has been approved by the National Center for Advancing Science (NCATS) and utilized by RUN. The Discussion Forums are created with the goal of advancing CTSA Program objectives in high priority areas of clinical and translational science. RUN actively engages members through in depth scheduled monthly meeting discussions with various relevant topics regarding the development and evaluation of clinical trials metrics, benchmarks, and scholarly publication and presentation activities. RESULTS/ANTICIPATED RESULTS: Topics covered in RUN monthly meetings include research units general budget guidelines, staff recruitment and retainment strategies, EPIC use in scheduling CRU research visits, and PPE for investigational drugs in context of USP800 requirements. RUN members vary in geographic location, type of clinical research (outpatient vs inpatient), resources, and research subject volume. They are engaged in online discussion and learning opportunities to improve translational science practices. A recent article titled “Impact of COVID-19 on Clinical Research Units (CRUs)” in JCTS is an example of best practices learned by RUN members and shared with the broader research community. DISCUSSION/SIGNIFICANCE: RUN as a Learning Research System enhances clinical and translational research unit capacity and efficiency, encouraging collaboration to contribute with improving public health. This network is aligned with the CTSAs mission of developing innovative solutions to improve translational science.

OBJECTIVES/GOALS: The study aims to explore the role of microglial behavior in cognitive impairment associated with vascular dementia (VaD) and long COVID. Using immunohistochemistry (IHC) and quantitative PCR (qPCR), we will assess the expression of Iba-1, a microglial activation marker, in subjects with VaD and SARS-CoV-2 infection. METHODS/STUDY POPULATION: Out of 48 female C57BL/6 mice, 24 had surgical intervention in the form of bilateral carotid artery stenosis (BCAS) for experimental induction of vascular dementia. After 2 weeks, 12 BCAS and 12 non-BCAS were infected with 1E4 PFU of mouse-adapted 10 (MA10) strain of SARS-CoV-2. 2 weeks post-infection, 4 weeks post-operatively, all animals were euthanized and tissues were processed for cDNA and histology. Immunofluorescence and RT-qPCR used to quantify microglia via Iba-1, BBB integrity via claudin-5 as well as occludin, GFAP, and integrin a5. RESULTS/ANTICIPATED RESULTS: We anticipate observing distinct patterns of microglial behavior in subjects with vascular dementia (VaD) and those with long COVID. Through immunohistochemistry (IHC), we expect to see increased Iba-1 expression, indicative of microglial activation. Quantitative PCR (qPCR) will likely corroborate these findings, showing elevated levels of Iba-1 mRNA. Lastly, we anticipate that the data will reveal interactions between microglia and the blood-brain barrier (BBB). These interactions could provide insights into how microglial behavior influences BBB integrity and, consequently, cognitive function in VaD and long COVID. DISCUSSION/SIGNIFICANCE: This study aims to clarify the role of microglia in cognitive decline linked to vascular dementia and long COVID. By categorizing patients based on microglial activation, we can better tailor treatments. The findings could lead to targeted therapies that address cognitive impairment in these conditions.

OBJECTIVES/GOALS: Large-scale clinical proteomic studies of cancer tissues often entail complex workflows and are resource-intensive. In this study we analyzed ovarian tumors using an emerging, high-throughput proteomic technology termed SWATH. We compared SWATH with the more widely used iTRAQ workflow based on robustness, complexity, ability to detect differential protein expression, and the elucidated biological information. METHODS/STUDY POPULATION: Proteomic measurements of 103 clinically-annotated high-grade serous ovarian cancer (HGSOC) tumors previously genomically characterized by The Cancer Genome Atlas were conducted using two orthogonal mass spectrometry-based proteomic methods: iTRAQ and SWATH. The analytical differences between the two methods were compared with respect to relative protein abundances. To assess the ability to classify the tumors into subtypes based on proteomic signatures, an unbiased molecular taxonomy of HGSOC was established using protein abundance data. The 1,599 proteins quantified in both datasets were classified based on z-score-transformed protein abundances, and the emergent protein modules were characterized using weighted gene-correlation network analysis and Reactome pathway enrichment. RESULTS/ANTICIPATED RESULTS: Despite the greater than two-fold difference in the analytical depth of each proteomic method, common differentially expressed proteins in enriched pathways associated with the HGSOC Mesenchymal subtype were identified by both methods. The stability of tumor subtype classification was sensitive to the number of analyzed samples, and the statistically stable subgroups were identified by the data from both methods. Additionally, the homologous recombination deficiency-associated enriched DNA repair and chromosome organization pathways were conserved in both data sets. DISCUSSION/SIGNIFICANCE OF IMPACT: SWATH is a robust proteomic method that can be used to elucidate cancer biology. The lower number of proteins detected by SWATH compared to iTRAQ is mitigated by its streamlined workflow, increased sample throughput, and reduced sample requirement. SWATH therefore presents novel opportunities to enhance the efficiency of clinical proteomic studies.

OBJECTIVES/GOALS: In 2021, Duke University expanded the myRESEARCHsuite (MRS) of research support services with the launch of myRESEARCHpath (MRP), an interactive roadmap for navigating the project lifecycle. MRP integrates with the existing MRS services, which include a personalized research portal (myRESEARCHhome) and team of experts (myRESEARCHnavigators). METHODS/STUDY POPULATION: MRP was developed as a collaborative effort to centralize essential research-related information across Duke University into one location. MRP provides a web-based platform to integrate policies, processes, and resources from over 40 research support offices, organized into topic-based pages throughout the project lifecycle. Each topic-based page provides integrated guidance, categorized related resources, and contact information for personalized support from subject matter experts. Additional features of MRP include a curated search function, and filters that refine the topic-based pages and related resources to only those applicable to selected project inclusions and organizational unit. RESULTS/ANTICIPATED RESULTS: Since the launch of MRP in January 2021 through the third quarter of 2021, 5,947 unique users accessed MRP for a total of 17,452 sessions. The most commonly accessed topic-based pages during this time period were: Activity disclosures (Other Support and Current and Pending) – 3,231 pageviews Animal welfare – 1,882 pageviews Proposal review and submission – 1,306 pageviews NIH research grants (R series) – 686 pageviews Proposal planning – 669 pageviews The most frequently searched terms (including spelling variants) were Other Support, Biosketch, NIH, and no-cost extensions. DISCUSSION/SIGNIFICANCE: This data suggests users are accessing MRP for guidance on new or recently updated requirements. Maintaining clear, unified, and current site content should be prioritized to continue emphasizing MRP as a central location for research-related information. Duke also plans to explore further integration of MRP with the other MRS services.

OBJECTIVES/GOALS: Glioblastomas (GBMs) are heterogeneous, treatment-resistant tumors that are driven by populations of cancer stem cells (CSCs). In this study, we perform an epigenetic-focused functional genomics screen in GBM organoids and identify WDR5 as an essential epigenetic regulator in the SOX2-enriched, therapy resistant cancer stem cell niche. METHODS/STUDY POPULATION: Despite their importance for tumor growth, few molecular mechanisms critical for CSC population maintenance have been exploited for therapeutic development. We developed a spatially resolved loss-of-function screen in GBM patient-derived organoids to identify essential epigenetic regulators in the SOX2-enriched, therapy resistant niche. Our niche-specific screens identified WDR5, an H3K4 histone methyltransferase responsible for activating specific gene expression, as indispensable for GBM CSC growth and survival. RESULTS/ANTICIPATED RESULTS: In GBM CSC models, WDR5 inhibitors blocked WRAD complex assembly and reduced H3K4 trimethylation and expression of genes involved in CSC-relevant oncogenic pathways. H3K4me3 peaks lost with WDR5 inhibitor treatment occurred disproportionally on POU transcription factor motifs, required for stem cell maintenance and including the POU5F1(OCT4)::SOX2 motif. We incorporated a SOX2/OCT4 motif driven GFP reporter system into our CSC cell models and found that WDR5 inhibitor treatment resulted in dose-dependent silencing of stem cell reporter activity. Further, WDR5 inhibitor treatment altered the stem cell state, disrupting CSC in vitro growth and self-renewal as well as in vivo tumor growth. DISCUSSION/SIGNIFICANCE: Our results unveiled the role of WDR5 in maintaining the CSC state in GBM and provide a rationale for therapeutic development of WDR5 inhibitors for GBM and other advanced cancers. This conceptual and experimental framework can be applied to many cancers, and can unmask unique microenvironmental biology and rationally designed combination therapies.

ABSTRACT IMPACT: Lack of regulatory knowledge and education is a key barrier to the translation of medical devices and we describe the design and results for a university graduate-level course providing training on medical device regulatory submissions for approval that can help fill this unmet need and improve and accelerate translational success. OBJECTIVES/GOALS: Within the Indiana CTSI, the Medical Technology Advance Program (MTAP) in the Purdue University Weldon School of Biomedical Engineering (BME) offers three courses in regulatory science and regulatory affairs for medical devices. One course is focused on regulatory submissions for approval, and this report details the course design and evaluation. METHODS/STUDY POPULATION: For Fall 2020, the Regulatory Submissions for Approval course was enhanced to increase participation from regulatory professionals in US FDA and industry, with the core content, curriculum and course design led by BME faculty. The course was taught two days per week and included both in-person and remote (synchronous or asynchronous) attendance options. During the first class session each week a topic was covered in standard lecture format by BME faculty with industry regulatory experience. During the second class session, guests from both industry and FDA were invited to provide in-depth discussion on the topic, share perspectives and viewpoints, present real-world examples, experiences, and case studies, and answer student questions. An end of semester survey evaluated the effectiveness of the course design. RESULTS/ANTICIPATED RESULTS: Medical Device regulatory submissions and related activities were taught including product classification, presubmissions and meetings, 510(k), de novo, EUA, PMA, HDE, and advisory panels. FDA history, regulatory careers, regulatory science, and EU, China, and Japan regulations were also discussed. Overall, 29 speakers from FDA and industry participated live via video calls. A survey completed by 21/23 studentsrevealed overall satisfaction: all reported increased regulatory understanding and 20/21 learned ‘a lot’ or ‘an incredible amount’. The weekly lecture was the top factor contributing to learning, and guest speakers were the next most important factors. Nearly all students indicated FDA and industry speakers were ‘very’ or ‘extremely’ valuable/helpful. Additional results will be presented. DISCUSSION/SIGNIFICANCE OF FINDINGS: The three courses are designed to improve medical device translation by training students to better understand regulatory processes and pathways. Survey results and feedback indicated this course was successful. Continued participation from FDA and industry is critical to the learning. Additional case studies will also help enhance learning.

OBJECTIVES/GOALS:

• Development of gold nanoparticles covalently linked to a photosensitizer for use to enhance radiation therapy. The particles will be thoroughly characterized structurally and mechanistically. The gold particles should enhance radiation activity by closer proximity to the photosensitizer and by increasing particle accumulation in the tumor.

METHODS/STUDY POPULATION:

• Gold nanoparticles were synthesized and coated with amine-terminated poly(ethylene) glycol, then covalently conjugated to chlorin e6, a known FDA-approved photosensitizer. The system was characterized using UV-Vis spectroscopy, transmission electron microscopy, and nanoparticle tracking analysis. The generation of reactive oxygen species was measured after X-irradiation. Enhanced cell killing was evaluated clonogenically in addition to assessment of in vivo efficacy and tumor pathology.

RESULTS/ANTICIPATED RESULTS:

• Conjugation of the particle to the photosensitizer was achieved, and the molecule was detected by UV-Vis spectroscopy. TEM and NTA showed no aggregation of the particles, and an increase in reactive oxygen species generation was observed. The conjugates increased cell killing during radiation treatment, whereas neither the particle alone nor the photosensitizer significantly affected clonogenic survival at the same concentrations. Breast tumors grown in immunocompetent mice showed increased necrotic tissue after a single 20 gy treatment in the presence of the conjugate.

DISCUSSION/SIGNIFICANCE OF IMPACT: Radiation therapy is widely used clinically, but dosage is limited largely to prevent injury to adjacent normal tissue. By increasing the local effect of radiation therapy, our gold conjugate has the potential to augment the effective radiation dose in the tumor, thereby reducing damage to healthy tissue and providing a more effective therapy.

OBJECTIVES/SPECIFIC AIMS: Black women experience discrimination that targets their intersecting gender and ethnic identities, termed gendered racism (Essed, 1991). The gendered racism Black women experience has been linked to negative mental health outcomes (Thomas etal., 2008). The ‘strong Black woman’ is a cultural symbol of strength depicting the Black woman as unwavered by hardships, such as gendered racism (Shorter-Gooden & Washington, 1996). However, recent research suggests that belief in the strong Black woman cultural construct is associated with negative mental health outcomes (Watson & Hunter, 2015). The goals of the current study were to (1) replicate previous findings suggesting that experiences with gendered racism is positively correlated with psychological distress, (2) replicate previous findings suggesting that belief in the strong Black woman construct is positively correlated with psychological distress, and (3) explore how experiences with gendered racism and belief in the strong Black woman construct might interact to predict distress. METHODS/STUDY POPULATION: A national sample of 112 Black women completed an online survey via MTurk. Survey measures included the Gendered Racial Microaggressions Scale, Strong Black Woman Cultural Construct Scale, and Psychological Distress Scale. RESULTS/ANTICIPATED RESULTS: Pearson correlation revealed that experiences with gendered racism was positively correlated with psychological distress, r = 0.23, p = .02. Pearson correlation also revealed that belief in the strong Black woman cultural construct was positively correlated with psychological distress, r = 0.39, p < .001. Multiple linear regression revealed an interaction between experiences with gendered racism and belief in the strong Black woman construct (β = -0.18, p = .04) that predicted psychological distress, R2 = .20, F(3,108) = 8.63, p < .01. Namely, for those with high belief in the strong Black woman construct, experiences with gendered racism did not predict distress, β = -0.31, t = -0.29, p = .78. However, for those with low belief in the construct, experiences with gendered racism positively predicted distress, β = -2.57, t = 2.31, p = .02. DISCUSSION/SIGNIFICANCE OF IMPACT: The results underscore the harmful effects of gendered racism and gendered racial stereotypes on Black women’s mental health outcomes. Striving to appear as the strong Black woman is not likely to help Black women overcome daily hardships. In fact, belief in the strong Black woman construct is likely to add extra difficulties.