OBJECTIVES/GOALS: This study aims to develop a method to examine whether blood-borne CNS-EV metal cargoes can serve as reliable biomarkers of CNS metal load and reveal a link between metal load and ALS development (i.e., neurodegenerative disease development). METHODS/STUDY POPULATION: CNS-EVs were isolated from human blood and plasma samples via direct immunoprecipitation using biotinylated antibodies for proteins known to be expressed in neurons (Contactin-2, i.e.) and astrocytes (glial-glutamate-aspartate-transporter, i.e., GLAST). Once isolated, protein concentrations in the EV samples were analyzed via ELISA assay, EV abundance was measured using ViewSizer Nanoparticle Tracking analysis, and EVs were visualized via Transmission Electron Microscopy. EVs were then analyzed for metal contents using a Perkin-Elmer NexION 350S via an ICP-MS/MS dynamic reaction cell method. RESULTS/ANTICIPATED RESULTS: Preliminary results demonstrate that it is feasible to quantify the metal contents of these CNS-derived EVs, particularly in terms of toxic metals known to be associated with neurodegenerative disorders, including copper, zinc, lead, aluminum, manganese, and iron. DISCUSSION/SIGNIFICANCE: CNS-derived EVs isolated from peripheral blood draws show promise as a potential biomarker of real-time metal load in the brain and spinal cord, with promising applications in predicting future development of neurodegenerative disorders (i.e., ALS) among patients with relevant elevated CNS metal loads.

OBJECTIVES/GOALS: The purpose of this study was to develop behavioral economic purchase tasks to assess interest in CM as a function of treatment cost and perceived effectiveness of CM as a function of abstinence incentive size in alcohol drinkers. Additionally, these purchase tasks are being assessed for their clinical utility in an ongoing clinical trial. METHODS/STUDY POPULATION: Alcohol drinkers recruited from Amazon Mechanical Turk completed behavioral economic purchase tasks measuring demand for CM based on targeted abstinence intervals and treatment effectiveness and alcohol use disorder severity assessments. Nonlinear mixed effects modeling was used to fit demand curves and assess the relationship between individual characteristics and demand metrics for CM. Ongoing analyses involve administering the same behavioral economic purchase tasks in heavy alcohol users in the ongoing clinical trial, which is aimed at reducing alcohol use through remotely implemented CM. RESULTS/ANTICIPATED RESULTS: Mechanical Turk participants reported higher probability of abstinence when offered larger incentives and required larger incentives when duration of abstinence required to earn the incentive was increased. Additionally, willingness to pay for treatment increased as effectiveness of treatment increased. It is anticipated that these patterns will be observed in the clinical trial participants. DISCUSSION/SIGNIFICANCE: Abstinence interval and treatment effectiveness are important features to consider when developing effective CM for widespread use, as these variables affected participants likelihood of being abstinent and their interest in treatment. We are currently working on verifying the results of these assessments in clinical trial participants.

OBJECTIVES/GOALS: This poster will explore lesion-symptom mapping and resting state functional magnetic resonance imaging (rsfMRI) in participants with aphasia. This analysis aims to reveal the behavioral impact of distinct lesions to the left inferior frontal gyrus (LIFG) and the potential for rsfMRI to provide evidence of cortical reorganization. METHODS/STUDY POPULATION: The analyses will be conducted on a small sample of approximately six participants with post-stroke language impairment (i.e., aphasia) recruited as part of an ongoing study. Lesion load in the anterior LIFG (aLIFG), consisting of the orbital (BA 47) and triangular (BA 45) parts, and the posterior LIFG (pLIFG), consisting of the opercular part (BA 44) will be correlated with performance on tasks of semantic and phonological processing. Functional connectivity will be determined between regions of interest (i.e., anterior superior temporal, posterior middle temporal, angular, and supramarginal gyri) in the left and/or right hemispheres based on rsfMRI data. RESULTS/ANTICIPATED RESULTS: Lesion load in the aLIFG is expected to correlate to performance on the semantic task, whereas lesion load in the pLIFG is expected to correlate to performance on the phonological task. For participants whose behavioral performance exceeds expectations based on lesion load (e.g., larger aLIFG lesion load, yet better semantic task performance), rsfMRI connectivity is expected to be greater (e.g., between the posterior middle temporal and angular gyri), suggesting cortical reorganization. DISCUSSION/SIGNIFICANCE: If our hypotheses are supported, our results will corroborate evidence of a functional spectrum (i.e., semantic-phonological) in the LIFG. Future work will compare rsfMRI connectivity to neurotypical controls and sites of connectivity may be used as targets in studies of non-invasive brain stimulation as an adjuvant to behavioral therapy.

OBJECTIVES/GOALS: Speech production requires mapping between sound-based and motor-based neural representations of a word – accomplished by learning internal models. However, the neural bases of these internal models remain unclear. The aim of this study is to provide experimental evidence for these internal models in the brain during speech production. METHODS/STUDY POPULATION: 16 healthy human adults were recruited for this electrooculography speech study. 20 English pseudowords were designed to vary on confusability along specific features of articulation (place vs manner). All words were controlled for length and voicing. Three task conditions were performed: speech perception, covert and overt speech production. EEG was recorded using a 64-channel Biosemi ActiveTwo system. EMG was recorded on the orbicularis orbis inferior and neck strap muscles. Overt productions were recorded with a high-quality microphone to determine overt production onset. EMG during was used to determine covert production onset. Neuroimaging: Representational Similarity Analysis (RSA), was used to probe the sound- and motor-based neural representations over sensors and time for each task. RESULTS/ANTICIPATED RESULTS: Production (motor) and perception (sound) neural representations were calculated using a cross-validated squared Euclidean distance metric. The RSA results in the speech perception task show a strong selectivity around 150ms, which is compatible with recent human electrocorticography findings in human superior temporal gyrus. Parietal sensors showed a large difference for motor-based neural representations, indicating a strong encoding for production related processes, as hypothesized by previous studies on the ventral and dorsal stream model of language. Temporal sensors, however, showed a large change for both motor- and sound-based neural representations. This is a surprising result since temporal regions are believed to be primarily engaged in perception (sound-based) processes. DISCUSSION/SIGNIFICANCE: This study used neuroimaging (EEG) and advanced multivariate pattern analysis (RSA) to test models of production (motor-) and perception (sound-) based neural representations in three different speech task conditions. These results show strong feasibility of this approach to map how the perception and production processes interact in the brain.

OBJECTIVES/GOALS: To develop a loop-mediated isothermal amplification (LAMP) assay for the detection of cytomegalovirus (CMV) infection and drug resistance. METHODS/STUDY POPULATION: We designed core and loop primers sets utilizing the NEB LAMP Primer Design Tool. Each set contained four or six primers targeting major-immediate early genes – essential for viral entry/replication – or regions known to confer resistance to the antiviral drug ganciclovir. Optimization of reactions conditions was achieved employing DNA reference materials. Reactions were visualized through a change in color as amplification reactions accumulated. Successful reaction conditions were selected based on specific amplification products in less than 60 minutes. Limits of detection were evaluated as the main performance outcome. RESULTS/ANTICIPATED RESULTS: Genomic data were extracted and used to design a series of LAMP primers (48 total) that aimed to detect specific genomic regions of CMV. Using this strategy, we successfully designed and identified eight primer sets that showed high 100% sensitivity and 100 % specificity, when detecting > 1.00 x 10^5 copies/mL of CMV gDNA. We are in the process of characterizing a new set of primers to determine the diagnostic utility of a LAMP assay in detecting selected single-nucleotide mutations at the UL97 loci. The expected outcomes at completion include: (1) the identification of LAMP primers to detect drug-resistance mutants, (2) defining optimal conditions for successful reactions, and (3) determining limits of detection for subsequent validation with clinical specimens. DISCUSSION/SIGNIFICANCE: CMV infection remains one of the most dangerous infectious agents for immunocompromised hosts, newborns, and unborn children. This study will describe a proof-of-concept LAMP assay for the genotypic detection of drug resistance in CMV-infected individuals and hence, create new avenues for selection of effective therapies to treat CMV disease.

OBJECTIVES/GOALS: Our goal is to develop a cost-effective approach for precision medicine treatment by providing computational predictions for new uses of currently available FDA approved, and experimental drugs for NSCLC. METHODS/STUDY POPULATION: Cell Lines: A549 (ATCC- CCL-185) Human epithelial Lung Carcinoma cells, H1792 (ATCC-CRL-5895) Human Lung Carcinoma cells. In Vitro Cytotoxicity Assay: A Vybrant® MTT Cell Proliferation Assay was used. Colony Formation Assay: NCI-H1792, A549 cells were seeded at a density of 500 cells/ dish, then treated with ARS-1620, Osimertinib. The Computational Analysis of Novel Drug Opportunities (CANDO):  Herein, we employed the bioanalytic docking (BANDOCK) protocol within CANDO to calculate the compound-protein interaction scores for a library of 13,218 compounds from DrugBank against a library of 5,317 protein structures from the Protein Data Bank, resulting in a proteomic interaction signature for each compound, and identified Osimertinib as the most likely EGFR/ErbB inhibitor to synergize with ARS-1620. RESULTS/ANTICIPATED RESULTS: ARS-1620 and Osimertinib in combination displays potent anti-tumor activity as evident by a decrease in cell viability with cytotoxicity assays, as well as reduced number of colonies in the colony formation assay for both A549 and H1792 cells. By using CANDO, and cross-referencing the obtained rankings with known experimental information, we have obtained drug predictions within the context of precision medicine. Our preliminary data indicates that EGFR inhibitor Osimertinib may be most structurally similar to KRAS G12C inhibitors overall, compared to other ErbB/ EGFR inhibitors. Validations with human cancer cell lines A549 and H1792 have confirmed that Osimertinib in combination with KRAS G12C inhibitor ARS-1620 may exhibit a synergistic effect in decreasing cellular proliferation and colony formation. DISCUSSION/SIGNIFICANCE: This suggests that this innovative drug combination therapy may help improve treatment outcomes for KRAS G12C(H1792) and KRASG12S(A549) mutant cancers. Cell migration and cell invasion studies in response to treatment with Osimertinib and ARS-1620 are currently ongoing.

OBJECTIVES/GOALS: Recent studies suggest nulliparous patients benefit from low dose aspirin (LDA), yet there are limited studies examining MFM providers adherence to USPSTF guidelines and predictors of adherence. We identified demographic, obstetric, and clinical characteristics associated with guideline concordant counseling on LDA in nulliparous women. METHODS/STUDY POPULATION: Retrospective cohort study of pregnant nulliparous patients who received MFM prenatal care at a single tertiary center (1/1/2019-6/30/2020). Multiple gestations, > 2 spontaneous or therapeutic abortions, and contraindications to LDA were excluded. Maternal demographic and clinical characteristics were abstracted from the electronic medical record. The primary outcome was documented LDA counseling based on USPSTF guidelines. Data were analyzed using bivariate analysis and logistic regression using R 4.1.0 (R Core Team, 2021). RESULTS/ANTICIPATED RESULTS: Among 394 records in the analysis cohort, 316 met USPSTF guidelines for LDA. 164 (51.9%) meeting guidelines had documented LDA counseling. 67.4% of individuals with ?1 major USPSTF risk factors were counseled and 50.7% with ?2 moderate risk factors were counseled (Table 1). Age at the estimated due date (EDD), Black or Other race, chronic hypertension, and obesity are significantly associated with higher odds of aspirin counseling (Table 2). Patients with chronic hypertension had 4.15 higher odds of receiving low dose aspirin counseling compared to non-hypertensive patient (Table 2). DISCUSSION/SIGNIFICANCE: Our results suggest that only 51.9% of patients eligible for LDA received counseling despite MFM care. Increasing MFM provider awareness about the USPSTF guidelines and creating tools that facilitate guideline concordant counseling may increase the number of eligible patients who are counseled about LDA.

OBJECTIVES/GOALS: The COVID-19 pandemic may have affected the relationship/experience of the mother-child dyad (Provenzi et al., 2021). Our objective is to identify the association of pandemic related maternal stress with child development. We will further evaluate the role of bonding, attachment, and trauma on this association. METHODS/STUDY POPULATION: We aim to recall a prospective cohort (n=200) of Latinx/Hispanic mothers from an ongoing study, power analysis will estimate minimum sample size (power=0.80 and alpha =0.05). Assessments of pandemic related maternal stress (PRMS) will be done with the COVID-19 and Perinatal Experiences Interview, perceived stress scale, and Parental Stress Index. Bonding, attachment, and trauma history will be assessed with psychological questionnaires and Childs behaviors with the Ackerman-CBCL questionnaire. Descriptive statistical analysis will be done. Correlations will identify associations and multivariate models will assess the role of parental bonding and effects of maternal attachment/trauma on associations to PRMS and child behavioral outcomes (controlling for confounding effects). RESULTS/ANTICIPATED RESULTS: First, we expect to find that mothers will report higher levels of stress (pandemic related, perceived, and parental) which will be associated with less bonding behaviors towards her child. Second, we expect that mothers levels of PRMS will be mediated by poorer bonding characteristics thus leading to negative child behavioral outcomes (i.e., poor regulation, crying spells, alterations in physiological patterns, and social-emotional developmental outcomes). Further mothers insecure attachment traits and trauma history will moderate perception of stress and negative child behavioral outcomes. DISCUSSION/SIGNIFICANCE: Results will describe stress in Latinx/Hispanics mothers during the pandemic and effects on child development. Identifying the role of maternal bonding/attachment will point to how this formative relationship has transformed during the pandemic, providing knowledge of mother-child resiliency.

OBJECTIVES/GOALS: Accumulating evidence supports the involvement of immune and inflammatory pathways in Bipolar Disorder (BD) pathophysiology. This pilot study aims to determine if altered peripheral IL-2, TNF-a, IL-4, IL-6, IL-10, IFN-y, IL-17A levels are associated with BD across mood episodes (euthymic, manic, depressive), and worsen neurocognitive function. METHODS/STUDY POPULATION: Twenty-eight participants (17 cases and 11 controls) were recruited. We assessed the clinical features and cytokine plasma levels of participants. Cytokines were measured using Flow Cytometry. All subjects were interviewed by a trained psychiatrist. Each participant was fasting before the blood sample was taken. Neuropsychological tests were used to measure verbal fluency, speed processing, working memory/attention, visuospatial skills, verbal learning, executive functions, and motor skills. Descriptive statistics were used to calculate the demographic characteristics of the sample. An independent-sample Kruskal-Wallis Test and Mann-Whitney Test were carried out using SPSS version 21. RESULTS/ANTICIPATED RESULTS: Serum biomarker concentration showed a decrease in levels of IL-4 (anti-inflammatory) in BD patients vs healthy controls (p < .05). There was a major concentration of IL-6 (pro-inflammatory) on bipolar patients vs controls (p = .003). When we analyze the results with the mood episodes, we found that patients with bipolar depression showed decreased levels of IL-4 (p = .046) and increase levels of IL-6 (p = .020) in comparison to the manic or euthymic episodes. In the neurocognitive tests, we found that the control participants had better performance in the working memory domain (p = .038) and also in the general performance (p = .036) in comparison to bipolar patients. We found also a positive significant correlation between IL-4 and verbal learning in the control sample (.829, p = .003). DISCUSSION/SIGNIFICANCE: The findings evidence a significant immune activation in bipolar patients, in particular during the depressive episode. Participants with BD have a decrease in the protective levels of IL-4 combined with high levels of IL-6 when compared to healthy controls. Worse neurocognitive functioning was found in bipolar patients.

OBJECTIVES/GOALS: Assess the diagnostic yield and test utilization of WES in patients having complex traits. We aim to evaluate the use of the first genetic approach for the identification of primary variants that contribute to neurogenetic disease etiology and influence onset and progression in Puerto Ricans. METHODS/STUDY POPULATION: Prospective cohort of 45 Puerto Rican probands (19 months - 36 years old) with complex neurogenetic traits that underwent WES (2019 - 2021). WES was performed, including copy number variant analysis and mitochondrial genome sequencing. We evaluated several factors possibly influencing the rate of WES diagnosis including early age, consanguinity, and family history of neurogenetic diseases. In addition, we only evaluated probands rather than dyads/trios and the clinical phenotypes. Descriptive analysis was performed, including a catalog of all variants reported. Multivariate analysis was performed to estimate the statistical association between variants and phenotypes reported and adjusting for potential confounders (age, sex, family history, income, health insurance and zip code). RESULTS/ANTICIPATED RESULTS: Auspiciously, positive pathogenic findings altered the clinical management in 29% of the probands in this study. A likely genetic diagnosis was achieved in 53% of the probands including pathogenic, likely pathogenic and variants of uncertain significance. Intronic variants, copy number variants detection and mitochondrial genome was included in WES methodology. Despite these facts, a 47% of the reported WES were negative, which deserve re-analysis potentially genotype based. Multivariate analysis is expected to adjust for potential confounders to establish a genotype-phenotype correlations in neurogenetic complex traits in this Puerto Rican admixed population. DISCUSSION/SIGNIFICANCE: Clinical WES offers an alternative approach for identification of variants in patients with complex traits. WES is also applicable in genetically heterogeneous individuals when specific genetic tests are not available or unsuccessful. Variants reported contribute to understand complex neurogenetic disease in underrepresented Puerto Ricans.

OBJECTIVES/GOALS: As the number of older adults (≥65 years) with T1D grows, there are limited data to guide care. In a six-month trial, CGM reduced hypoglycemia in older adults, yet there are challenges for widespread uptake. Our objective is to characterize older adults experiences with using CGM and define suboptimal responses signaling a need for resources or support. METHODS/STUDY POPULATION: The study will engage key stakeholders (i.e., older adults with T1D, caregivers [recruited as patient-caregiver dyads], and providers [endocrinologists, geriatricians, diabetes educators]) for a Group Model Building (GMB). GMB is a participatory approach to system dynamics in which participants share perceptions and experiences with a problem and collaboratively explore the system structure that shapes those trends. A series of 8 GMB workshops will be held with 3-8 participants. The final study n will be determined by thematic saturation. Workshops comprise 1) a questionnaire, 2) a GMB session, and 3) a focus group discussion. GMB will follow a replicable process to generate a model of the complex web of causal determinants affecting CGM-related experiences, including optimal and suboptimal CGM responses. RESULTS/ANTICIPATED RESULTS: To date, the study has enrolled 33 participants, including 28 older adults living with T1D and 5 caregivers (mean age = 74 years, range 67-83 years). Twenty-four patient participants will be active CGM users and 4 will be CGM non-users. The study will report on patient data capture from the questionnaire and EMR, including demographics, experiences, familiarity, and confidence surrounding CGM use; diabetes duration; insulin pump use; history of severe hypoglycemia. Analysis of aggregated data will generate causal loop diagrams that integrate pertinent theoretical frameworks, lived experiences, and CGM outcomes. Maps will be used to identify a set of suboptimal CGM responses (i.e., key outcome trajectories) that signal a need for action, with a diagram of factors that interact to produce each response. DISCUSSION/SIGNIFICANCE: Delivering CGM to older adults with T1D demands new approaches. This study will yield critical evidence to tailor support and resources for effective CGM use in older adults. Findings may be translated into suite of pragmatic interventions to bolster CGM use and matched to individual patients expected to benefit using a precision medicine framework.

OBJECTIVES/GOALS: Several studies compare perioperative parameters and postoperative morphology of open cranial vault remodeling (CVR) and endoscopic-assisted craniectomy (EAC) in the treatment of sagittal craniosynostosis. However, data on neurodevelopmental outcomes comparing these techniques is lacking. METHODS/STUDY POPULATION: Patients with repaired sagittal synostosis were enrolled from a single tertiary care center between five and fifteen years of age. Patients with any craniofacial syndrome (e.g., 22q11.2 deletion) or other medical conditions that could affect neurodevelopment were excluded. Neurodevelopment was measured using the Differential Ability Scale-II (DAS-II). The General Cognitive Ability (GCA) score was the primary outcome measure, describing overall intellectual ability. Secondary outcome measures were measurements of cephalic index (CI) from pre- and postoperative imaging studies. Family socioeconomic status (SES) was estimated using the Hollingshead index (total score, continuous). RESULTS/ANTICIPATED RESULTS: 26 patients [10 CVR (2 Female:8 Male), 16 EAC (6 F:10 M)] were studied. Mean age at repair was greater with CVR than EAC (15.5 ± 8.5 months vs 3.5 ± 1.3 months, respectively, p < 0.001). Mean age at DAS-II testing was 9.0 ± 2.2 years. Patients GCA scores were within the average range relative to test norms (CVR 95.7 ± 15.4 vs EAC 102.6 ± 17.4). After adjusting for SES and preoperative CI, group differences in GCA were modest and statistically nonsignificant (p = 0.646). GCA scores were associated with SES (p = 0.054) but not preop CI (p = 0.479). Differences between CVR and EAC were not statistically significant for any of the imaging parameters (pre and postop CI, pre to postop change in CI, or age at postop imaging; 0.131 <= p <= 0.867). None of the independent variables were significantly associated with postop CI (0.140 <= p <= 0.689). DISCUSSION/SIGNIFICANCE: Interim analysis of the preliminary data suggests no association between surgical procedure and cognitive and morphological outcomes at school-age in patients with sagittal craniosynostosis who underwent CVR versus EAC. Collection of data continues, with the goal to enroll 50 participants in each group by 2022.

OBJECTIVES/GOALS: Approximately 10% of COVID-19 patients experience multiple symptoms weeks and months after the acute phase of infection. Our goal was to use advanced machine learning methods to identify PASC phenotypes based on their symptom profiles, and their association with critical adverse outcomes, with the goal of designing future targeted interventions. METHODS/STUDY POPULATION: Data. All COVID-19 outpatients from 12 University of Minnesota hospitals and 60 clinics. Independent variables consisted of 20 CDC-defined PASC symptoms extracted from clinical notes using NLP. Covariates included demographics, and outcomes included New Psychological Diagnostic Evaluation, and Number of PASC Hospital Visits (>=5). Cases (n=3235) consisted of patients with at least one symptom, and controls (n=3034) consisted of patients with no symptoms. Method. (1) Used bipartite network analysis and modularity maximization to identify patient-symptom biclusters. (2) Used multivariable logistic regression (adjusted for demographics and corrected through Bonferroni) to measure the odds ratio of each patient bicluster to adverse outcomes, compared to controls, and to each of the other biclusters. RESULTS/ANTICIPATED RESULTS: The analysis identified 6 PASC phenotypes (http://www.skbhavnani.com/DIVA/Images/Fig-1-PASC-Network.jpg), which was statistically significant compared to 1000 random permutations of the data (PASC=.31, Random Median=.27, z=11, P<.01). Three of the clusters (Cluster-1, Cluster-4, and Cluster-5 encircled with ovals in Fig. 1) contained CNS-related symptoms, which had statistically significant risk for one or both of the adverse outcomes. For example, Cluster-1 with critical CNS symptoms (depression, insomnia, anxiety, brain-fog/difficulty-thinking), had a significantly higher OR compared to the controls for New Psychological Diagnostic Evaluation (OR=6.6, CI=4.9-9.1, P-corr<.001), in addition to having a significantly higher ORs for the same outcome compared to all the other clusters. DISCUSSION/SIGNIFICANCE: The results identified distinct PASC phenotypes based on symptom profiles, with three of them related to CNS symptoms, each of which had significantly higher risk for specific adverse outcomes compared to controls. We will test whether these phenotypes replicate in the N3C data, and explore their translation into triage and treatment strategies.

OBJECTIVES/GOALS: The aim of this study is to examine the prevalence of communication sources about genetic testing among adult women with a higher risk of cancer in the U.S. and identify factors associated with these communication sources by analyzing the Health Information National Trends Survey (HINTS; 2017-2020) data, a large, nationally representative sample. METHODS/STUDY POPULATION: HINTS aims to gain knowledge about the use of and access to cancer information by the public. Used data was collected between 2017 and 2020 from a sample of adult participants who self-reported as female, aged 18+, having no personal cancer diagnosis, and having a family history of cancer. The primary outcome measure is source of communication about genetic testing. Some categories are the media, family members, or a healthcare professional. Analytical approaches include the Pearsons Chi-squared test and poisson regression model to estimate prevalence ratios and examine the association between sociodemographic characteristics and sources of communication, adjusting for clinical and health belief factors as covariates. RESULTS/ANTICIPATED RESULTS: This study is in progress. It is anticipated that the most overall prevalent method of communication about genetic testing will be via the media. In multivariate models, it is anticipated that women who are younger, Black, have a lower education, have lower income, and no health insurance are more likely to receive communication about genetic testing from a source other than a health professional or not at all. After adjusting for clinical and health belief factors such as co-morbid conditions, having a primary care physician, and general knowledge about genetic testing, there may be some association between the above mentioned sociodemographic factors and receiving communication about genetic testing via a healthcare professional. DISCUSSION/SIGNIFICANCE: Reporting on the association between sociodemographic factors and sources of communication can aid in an intervention design to better promote genetic testing. This can be most beneficial among vulnerable groups like Black women to better understand their own genetic risk of cancer and to make informed decisions about their health.

OBJECTIVES/GOALS: Asymmetries in lung pathophysiology can result in a maldistribution of gas between regions of the lungs which may generate dangerous pressures that are not observable by clinicians. Our study aims to demonstrate and quantify this through use of high-fidelity simulators to represent a range of commonly encountered clinical pathologies. METHODS/STUDY POPULATION: A benchtop study was performed with two high-fidelity breathing simulators, each representing one lung. This system allows for real-time monitoring of pressure and lung dynamics in a two-lung asymmetric injury model. One simulator was set to a fixed compliance and a resistance. A second simulator had a range of compliance and resistance values. Data were collected for 15 different test cases across a distribution of asymmetries. Each test case is run for 30 cycles. At the end of each ventilatory cycle, a short expiratory hold is performed, allowing pressure in the lung simulator, tubing, and ventilator circuit to equilibrate between cycles. RESULTS/ANTICIPATED RESULTS: Maldistribution of tidal volume was demonstrated when the compliance ratio between lung models (CL1/CL2) was 0.2 and the resistance ratio (RL1/RL2) was 10 with 23.9% (99% CI: 23.9-24.0%) of the gas volume distributed to lung 1 (103 mL L1 vs 327 mL in L2). Additionally, the injured lung when compared with the normal lung experienced higher peak pressures (12.8 cm H2O vs. 6.9 cm H2O, L1 and L2 respectively) and higher compartmentalized plateau pressures (11.5 cm H2O vs. 6.8 cm H2O, L1 and L2 respectively). DISCUSSION/SIGNIFICANCE: We demonstrate significant maldistribution of volume and pressures between two lungs in an asymmetric injury model. This study suggests significant impact of asymmetry in current lung-protective mechanical ventilation strategies and calls for better understanding of case-specific pathophysiologic changes affecting each of the two lungs.

OBJECTIVES/GOALS: Neonatal endotracheal tubes (ETTs) are usually uncuffed to avoid subglottic stenosis and other complications, but cuffed ETTs allow better ventilation. Our goal was to detect and control pressure in the cuff below the limit of occluding venous flow to minimize the risk of subglottic stenosis. METHODS/STUDY POPULATION: We designed a pressure sensor to fit on a 2.5 ETT for prototype testing in 8 age adult female rabbits. Eight uncuffed age- and sex- matched rabbits served as control. Study duration was 2 hours during which pressure in the cuff was limited by novel sensor (intervention) or auscultation (control). Anesthesia was maintained with sevoflurane. Ventilation was provided mechanically. Subsequently the tracheae were removed, sectioned crosswise, and compared histologically for mucosal damage. RESULTS/ANTICIPATED RESULTS: Preliminary data demonstrated an almost 30% greater amount of intact mucosa in the intervention group. The sensor also provided data on heart rate and respiratory rate, although this signal was not optimal. After filing an invention disclosure and provisional patent, we are refining our device to include multiple compartments for local control of cuff pressure and applying for a STTR Phase I/II application. DISCUSSION/SIGNIFICANCE: Ventilation in neonates with uncuffed ETTs can be suboptimal due to leak around the tube, but cuffed ETTs pose the threat of subglottic stenosis and other complications. We have designed a prototype cuffed ETT with a sensor to maintain low cuff pressure while preventing leaks and largely avoiding damage to the tracheal mucosa.

OBJECTIVES/GOALS: In a familial case where 10 of 17 members inherited EA/LVNC in an autosomal dominant pattern, we discovered a novel, damaging missense variant in the gene KLHL26 that segregates with disease and comprises an altered electrostatic surface profile, likely decoupling the CUL3-interactome. We hypothesize that this KLHL26 variant is etiologic of EA/LVNC. METHODS/STUDY POPULATION: We differentiated a family trio (a heart-healthy daughter and EA/LVNC-affected mother and daughter) of induced pluripotent stem cells into cardiomyocytes (iPSC-CMs) in a blinded manner on three iPSC clones per subject. Using flow cytometry, immunofluorescence, and biomechanical, electrophysiological, and automated contraction methods, we investigated iPSC-CM differentiation efficiency between D10-20, contractility analysis and cell cycle regulation at D20, and sarcomere organization at D60. We further conducted differential analyses following label-free protein and RNA-Seq quantification at D20. Via CRISPR-Cas9 gene editing, we plan to characterize KLHL26 variant-specific iPSC-CM alterations and connect findings to discoveries from patient-specific studies. RESULTS/ANTICIPATED RESULTS: All iPSC lines differentiated into CMs with an increased percentage of cTnT+ cells in the affected daughter line. In comparison to the unaffected, affected iPSC-CMs had fewer contractions per minute and altered calcium transients, mainly a higher amount of total calcium release, faster rate of rise and faster rate of fall. The affected daughter line further had shorter shortening and relaxation times, higher proliferation, lower apoptosis, and a smaller cell surface area per cardiac nucleus. The affected mother line trended in a similar direction to the affected daughter line. There were no gross differences in sarcomere organization between the lines. We also discovered differential expression of candidate proteins such as kinase VRK1 and collagen COL5A1 from proteomic profiling. DISCUSSION/SIGNIFICANCE: These discoveries suggest that EA/LVNC characteristics or pathogenesis may result from decreased contractile ability, altered calcium transients, and cell cycle dysregulation. Through the KLHL26 variant correction and introduction in the daughter lines, we will build upon this understanding to inform exploration of critical clinical targets.

OBJECTIVES/GOALS: To assess the impact of federal regulations and policies relating to disposable Electronic Nicotine Delivery Systems (ENDS) on youth consumption of these products by identifying factors enabling its growing consumption among youth users and its relation to adolescent addiction to nicotine. METHODS/STUDY POPULATION: Disposable ENDS are all-in-one devices with pre-filled nicotine liquid and a built-in battery. Recent data shows increased sales as users, including youth, are switching from pod-based to disposable ENDS. Thus, an understanding of the regulatory landscape for these products will provide insight on how to mitigate youth nicotine addiction. Data from the Centers for Disease Control and Prevention 2021 National Youth Tobacco Survey (NYTS) was analyzed for patterns of adolescent use. FDA statements and actions involving disposable ENDS companies were reviewed to evaluate the current FDA stance. Analyses of both data sets identified factors enabling the growth in sale of disposable ENDS. RESULTS/ANTICIPATED RESULTS: The NYTS reported 53.7% of youth ENDS users report using disposable ENDS and Puff Bar is the leading ENDS device among youth consumers. In March 2021, Puff Bar announced a return to market with “tobacco-free nicotine” after ceasing sales following an FDA warning letter in July 2020. But synthetic nicotine retains the same chemical properties as tobacco-derived nicotine and the same risks for addiction and abuse. The Food and Drug Administration (FDA) maintains synthetic nicotine products will be regulated on a case-to-case basis, suggesting “closed system devices” containing synthetic nicotine may not be regulated as tobacco products. DISCUSSION/SIGNIFICANCE: The growing popularity of disposable ENDS among youth is problematic. Awareness of strategic regulatory defiance (i.e., Puff Bar), will bring light to industry sales tactics. To develop comprehensive data on disposable ENDS use by young adults, an anonymous survey of college students will be conducted.

OBJECTIVES/GOALS: Review all approved companion diagnostics (CDxs) labeling for therapeutic class language. Examine the regulatory pathways of CDx products whose labels contain therapeutic class labeling. Develop recommendations for pharmaceutical industry professionals on best practices in the co-development of CDxs and oncology therapeutic products. METHODS/STUDY POPULATION: Literature discussing companion diagnostics was reviewed from EBSCOhost, PubMed, and OVID. The Intended Use language within CDx labels on the Food and Drug Administrations “List of Cleared or Approved Companion Diagnostic Devices (In Vitro and Imaging Tools)” website were reviewed on November 1, 2021 for therapeutics class language. For CDx products with therapeutic class label language, the regulatory history was evaluated to determine the development approach taken to achieve the language. RESULTS/ANTICIPATED RESULTS: A total of 45 CDxs were identified, of which only 2 contained therapeutic class labeling, both of which were devices for the identification of epidermal growth factor receptor (EGFR) mutations for the treatment of non-small cell lung cancer (NSCLC). Three additional EGFR CDxs were approved; however, they did not contain therapeutic class labeling. The first CDx was the Cobas EGFR Mutation Test V2, which received therapeutic group label language as an update on October 27, 2020; however, prior to the therapeutic class labeling, three oncology products were named in the Intended Use: Tarceva (erlotinib), Tagrisso (osimertinib), and Iressa (gefitinib). The second CDx to incorporate therapeutic class labeling was the ONCO/Reveal Diagnostic Lung and Colon Cancer Assay upon initial approval on July 30, 2021. DISCUSSION/SIGNIFICANCE: EGFR CDxs are the first to shift towards therapeutic class labeling. Indication, molecular alterations, and mechanism of action of the approved therapeutic class products, number of products approved, as well as CDx analytical and clinical validation influence class label relevance. Discussions with the FDA are encouraged early in development.

OBJECTIVES/GOALS: Antiretroviral treatments (ART) suppress retroviruses, like the human immunodeficiency virus (HIV). The goal is to understand how antiretroviral drug patents contribute to overpriced HIV medications, thereby causing cost-related treatment non-adherence and inhibiting viral suppression. METHODS/STUDY POPULATION: Currently, HIV affects 57,700 individuals in Los Angeles County (LAC). Data was compiled from Centers for Disease Control and Prevention (CDC), hiv.gov, lacounty.HIV, publichealth.lacounty.gov, and the U.S. Patent and Trade Office. A review of existing literature examined the role of ART patents on cost-related non-adherence and declining viral suppression on individuals living with HIV in LAC. Lastly, a comparison of HIV medication prices of expired and non-expired patents was conducted, indicating the effects of undue extensions of market exclusivity on ART regimen pricing, and how this affects treatment adherence. RESULTS/ANTICIPATED RESULTS: Of the ten ARTs examined, four had expired patents and six had active patents. Those with active patents cost more than those with inactive patents because active patent status prevents price reductions. Patent strategies–pay-for-delay settlements and patent evergreening–unduly extend market exclusivity, keeping ART at prohibitive costs and preventing generic competition. Individuals facing cost-related non-adherence were less virally suppressed at their last viral load test (64%) and at all tests during the year (54%). Thus, over-patented ARTs increase treatment prices, causing cost-related non-adherence to ART regimens. The implications include disease progression and less viral suppression. DISCUSSION/SIGNIFICANCE: The U.S. has the highest ART prices, yet the lowest rate of HIV viral suppression (54%) among all well-resourced countries. Undue extensions of market exclusivity cause ARTs to remain at prohibitive costs, preventing some patients from affording ART treatments, minimizing their viral suppression.