bluuebunny/crossref_metadata_2025 huggingface dataset:
271 Search Solutions: Investigation into CTSA Hubs COVID-19 Vaccine Information
Authors: Bashir Hasan M. , Tafuto Barbara
Published: April 2022

OBJECTIVES/GOALS: While CTSA hubs have contributed to the successes in COVID-19 clinical research, understanding the role of the CTSA consortium in translating vaccine efficacy and availability to awareness and implementation. The goal of this study is to quantitatively assess the use of social media in the dissemination of COVID 19 vaccine content across 60+ CTSA Hubs. METHODS/STUDY POPULATION: Structured search terms in the CTSA Search Solutions database were used to identify CTSA Hub website pages highlighting “COVID-19 Vaccination Information.” Each link identified was manually reviewed for vaccination content. The links and content identified by CTSA Search Solutions were then validated by advanced Google search operator “vaccine site” and manual review of CTSA Hub websites. Official CTSA Hub social media platforms were searched for vaccine proliferation content from January 1, 2020 to November 11, 2021. Data points collected included Community Targeted Vaccine Content, Vaccine Awareness, Vaccine Distribution, Vaccine Clinical Trial, Vaccine Related Media, Social Media Presence, and Social Media Vaccine Proliferation. RESULTS/ANTICIPATED RESULTS: In examining content, of the 64 listed CTSA Hubs, 52 of (67.2%) hosted one or more categories of COVID-19 vaccine information, and 16 (25%) hosted three or more categories. The most common category was “Community Targeted Information” with 27 (42.2%) hubs; the least common category was “Vaccine Distribution” with 5 (7.8%) hubs. Examining social media for vaccine proliferation, 41 (64.1%) hubs had connected social media and 23 (35.9%) hubs had vaccine proliferation posts. The most common platform was Twitter with 37 hubs; the least common platform was Instagram with 5 hubs. DISCUSSION/SIGNIFICANCE: Our investigation demonstrated varying Phase 4 translational efforts via social media among the CTSA Hubs for COVID-19 vaccine implementation. These novel findings identify promising opportunities for enhancement while emphasizing proven strategies from CTSA hubs nationwide. Further research will elucidate granular trends among CTSA hubs.
278 Developing AMILDA: An Intervention to Improve Symptoms and Medication Adherence among Latinx Adults with Depression and Anxiety
Authors: Mitrzyk Beatriz Manzor , Gonzalez Lizbeth , Alegria Margarita , Pinto Rogerio M , Farris Karen
Published: April 2022

OBJECTIVES/GOALS: To develop the Adherence to Medications In Latinx adults with Depression and Anxiety (AMILDA) intervention using community engaged research principles, AMILDA will integrate culturally tailored messaging to improve depression and anxiety symptoms and adherence to antidepressant medications. METHODS/STUDY POPULATION: Bilingual steering committee members representing primary care and behavioral health professionals from 2 primary care clinics, and 1 lay community member met monthly for 1 year to develop the intervention. One clinic is an academic health center and the other is a federally qualified health center. Members identified, developed, and voted on the interventions components, therapeutic goals, and the mechanisms underlying its effectiveness that needed to be studied. To evaluate AMILDAs components, we conducted an observational study of health care providers (N=13) and Latinx adults with depression and/or anxiety (patients; N=26) from both clinics, and community pharmacists (N=9) from surrounding pharmacies to gather feedback. Based on their input, we revised the intervention. RESULTS/ANTICIPATED RESULTS: AMILDA consists of culturally tailored collaborative drug therapy management with a collaborative practice agreement between the physician and pharmacist. Patients recently started on an antidepressant receive language and culturally concordant motivational interviewing, teach back, and infographics; AMILDA is designed to improve symptoms, adherence, and health determinants. Most provider, pharmacist, and patient participants reported that AMILDA could be beneficial. Several participants indicated a lack of understanding about a US pharmacists scope of practice. Recommendations included ensuing language concordance, incorporating more statistics and scientific data, using images that are relatable, avoiding idioms and stereotypes, taking the time to build rapport, and being friendly. DISCUSSION/SIGNIFICANCE: AMILDA will be tested in a pilot effectiveness randomized controlled study to determine if it improves depression and anxiety symptoms and adherence to antidepressants among Latinx adults compared with enhanced usual care. The mechanisms to be assessed include self-efficacy, antidepressant knowledge, and personal health literacy.
289 Helpline Services Before, During, and After the COVID-19 Pandemic: A Time Series Analysis
Authors: Cua Grace , Segovia David , Poole Jim , Gore Devyani , McGowan-Tomke Jennifer , James Alexa , Frank Ben , Atkins Marc
Published: April 2022

OBJECTIVES/GOALS: This study examined patterns in helpline call data as the COVID-19 pandemic evolved including the impact of stay-at-home orders, relaxing of restrictive orders, and stages of vaccine uptake, as well as differences in call volume by Chicago neighborhood health indicators. METHODS/STUDY POPULATION: From November 1, 2018 to June 30, 2021, 56 NAMI-Chicago workers accepted 26,173 helpline calls from 9,374 individuals from 438 zip codes across northeastern Illinois with the majority of calls from high poverty Chicago communities. Descriptive and time series analyses examined patterns in call volume related to the onset of the COVID-19 pandemic, Illinois Stay-at-Home Order, and Illinois reopening and vaccine uptake plan relative to comparable times the prior year. Health indicators from the Chicago Health Atlas (https://chicagohealthatlas.org/) were examined to determine patterns related to NAMI call volume and various health indicators at the zip code level. RESULTS/ANTICIPATED RESULTS: Time series analysis indicated the greatest number of calls occurred in 2020; specifically, there was a 212% increase in call volume and 331% increase in repeat callers (three or more calls per caller) during the first and second phase (March 20th to May 28th) of Illinois Stay-at-Home Order from 2019 to 2020. Analysis of the callers primary need indicated NAMI provided resources and referrals to people with unmet basic needs such as housing, food, and access to healthcare during the height of COVID-19 Pandemic in 2020. A series of ANOVAs indicated that individuals from Chicago zip codes with high levels of uninsured rates, poverty rates, households using SNAP benefits, and economic diversity called NAMI significantly more than those with low levels of these health indicators. DISCUSSION/SIGNIFICANCE: Helplines are a much-needed model to assess needs and implement services during public health crises, particularly in communities experiencing economic hardship and stress. Implications for behavioral health service needs both during and following the pandemic will be discussed.
297 The Acute to Chronic Pain Signatures Program (A2CPS): Conceptual Design and Protocol Implementation of a Multi-site Observational Study Assessing Risk and Resilience Biomarkers
Authors: Berardi Giovanni , Vance Carol GT , Dailey Dana L , Ecklund Dixie , Coffey Christopher , Bayman Emine , Sluka Kathleen A.
Published: April 2022

OBJECTIVES/GOALS: The A2CPS was funded by the National Institutes of Health (NIH) Common Fund to identify biomarkers and their collective biosignatures (combination of several biomarkers) that predict susceptibility or resilience to the development of chronic pain. METHODS/STUDY POPULATION: The A2CPS includes 2-Multisite Clinical Centers (10 recruitment sites and 6 data collection sites), 1-Clinical Coordinating Center, 1-Data Integration and Resource Center, 3-Omics Data Generation Centers, and representation from the NIH. The A2CPS will recruit a large cohort from 2 different surgical interventions, total knee arthroplasty (n?1800) and thoracotomy (n?1800). This observational study will collect candidate and exploratory biomarkers across the following domains: clinical pain, fatigue, function, sleep, psychosocial, genomics, proteomics, metabolomics, lipidomics, pain sensitivity, and brain imaging. Data will be collected before and up to 3 months after surgery to determine factors that predict chronic pain at 6 months. RESULTS/ANTICIPATED RESULTS: Recruitment started in 2021 following standard operating procedures and is ongoing at both Multisite Clinical Centers. The A2CPS will provide an example of collaborative, multidisciplinary efforts in establishing a data repository consisting of biopsychosocial markers that will be available to the research community to test novel hypotheses. This presentation will describe the conceptual design, study aims, biomarker selection, protocol standardization and study implementation for the A2CPS. An update on study progress and data completeness will be presented. Final results will be reported after study completion which is anticipated by 2024. DISCUSSION/SIGNIFICANCE: Identifying biomarkers and biosignatures that predict high- versus low-risk for the transition to chronic pain will inform future clinical trials, identify novel therapeutic targets, and advance personalized pain treatment strategies; ultimately transforming the prevention and treatment of chronic postsurgical pain.
309 MYC Inhibition Overcomes IMiD Resistance in Heterogeneous Multiple Myeloma Populations
Authors: Davis Lorraine , Walker Zachary J. , Ohlstrom Denis , Stevens Brett M. , Forsberg Peter A. , Mark Tomer M. , Jordan Craig T. , Sherbenou Daniel W.
Published: April 2022

OBJECTIVES/GOALS: Immunomodulatory drugs (IMiDs) are critical to multiple myeloma (MM) disease control. IMiDs act by inducing Cereblon-dependent degradation of IKZF1 and IKZF3, which leads to IRF4 and MYC downregulation (collectively termed the “Ikaros axis”). We therefore hypothesized that IMiD treatment fails to downregulate the Ikaros axis in IMiD resistant MM. METHODS/STUDY POPULATION: To measure IMiD-induced Ikaros axis downregulation, we designed an intracellular flow cytometry assay that measured relative protein levels of IKZF1, IKZF3, IRF4 and MYC in MM cells following ex vivo treatment with the IMiD Pomalidomide (Pom). We established this assay using Pom-sensitive parental and dose-escalated Pom-resistant MM cell lines before assessing Ikaros axis downregulation in CD38+CD138+ MM cells in patient samples (bone marrow aspirates). To assess the Ikaros axis in the context of MM intratumoral heterogeneity, we used a 35-marker mass cytometry panel to simultaneously characterize MM subpopulations in patient samples. Lastly, we determined ex vivo drug sensitivity in patient samples via flow cytometry. RESULTS/ANTICIPATED RESULTS: Our hypothesis was supported in MM cell lines, as resistant lines showed no IMiD-induced decrease in any Ikaros axis proteins. However, when assessed in patient samples, Pom treatment caused a significant decrease in IKZF1, IKZF3 and IRF4 regardless of IMiD sensitivity. Mass cytometry in patient samples revealed that individual Ikaros axis proteins were differentially expressed between subpopulations. When correlating this with ex vivo Pom sensitivity of MM subpopulations, we observed that low IKZF1 and IKZF3 corresponded to Pom resistance. Interestingly, most of these resistant populations still expressed MYC. We therefore assessed whether IMiD resistant MM was MYC dependent by treating with MYCi975. In 88% (7/8) of patient samples tested, IMiD resistant MM cells were sensitive to MYC inhibition. DISCUSSION/SIGNIFICANCE: While our findings did not support our initial hypothesis, our data suggest a mechanism where MYC expression becomes Ikaros axis independent to drive IMiD resistance, and resistant MM is still dependent on MYC. This suggests targeting MYC directly or indirectly via a mechanism to be determined may be an effective strategy to eradicate IMiD resistant MM.
312 Developing a Digitally Integrated Endotracheal Tube for Neonates to Improve Safety and Respiratory Function
Authors: Nienaber Thomas , Perez Sarah , Stephenson Krista , Sanford Joseph , Abella Adria , Jensen Morten , Sexton Kevin
Published: April 2022

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.
315 Development of a Mobile Software Application to Promote Education, Self-Care, and Treatment Follow-Up among Puerto Rican Breast Cancer Patients
Authors: Abreu Jenniffer , Soto-Torres Brenda , Reynoso-Jimenez Elan A. , Acevedo-Suárez Carlos A.
Published: April 2022

OBJECTIVES/GOALS: This project will design, develop and pilot-test a Spanish-mobile application for breast cancer patients. It is intended that patients will improve treatment adherence, have a personal information storage system for their follow-up healthcare and can have greater control in the processes of diagnosis, treatment, education, and recovery. METHODS/STUDY POPULATION: In the first phase, a team of undergraduate engineering students was recruited to work on the areas of: Front End, Database Development, Security Protocols, and Integration. Team Venus will provide the cancer-related content. In the second phase, the engineering team will identify bugs, usability, satisfaction, and workability of the application among a pilot convenience sample of 20 participants from four cancer clinics and the community. The inclusion criteria are being > 21 years old and having a cell phone. The exclusion criterion is not knowing how/being able to use a mobile application. Participants will complete a questionnaire on sociodemographic data, mobile phone and application use, efficacy, usability, and security. Univariate and bivariate statistical analysis will be performed using SPSS. RESULTS/ANTICIPATED RESULTS: The researchers are defining specific visual and content features they would like the application to have. They are also searching and collecting reliable information about cancer from primary sources to incorporate into the tools available to the application’s users. In addition, they have also begun to identify potential cancer treatment facilities and medical personnel to assist in the recruitment of patients for pilot-testing the application. The researchers have divided them into geographical areas in Puerto Rico and each Team Venus member will approach and orient patients and medical personnel about the project. DISCUSSION/SIGNIFICANCE: It is intended that patients will improve treatment adherence, have a storage system for their follow-up healthcare and greater control in their processes of diagnosis, treatment, education, and recovery. Future goals include use of the database feature by researchers and expanding the testing of the application with a larger patient-base.
320 Genetic Compensation as a mechanism underlying patients with Rare ALS
Authors: Gomez Adriana Morales , Staff Nathan , Ekker Stephen C.
Published: April 2022

OBJECTIVES/GOALS: Rare mutations in CHCHD10 gene are found in 1% of patients with familial Amyotrophic lateral sclerosis (ALS). The overall goal of this study is to utilize induced pluripotent stem cells (iPSCs) as an in vitro model organism for rare ALS variants to evaluate the mechanism of transcription adaptation of CHCHD10/2 as a potential therapeutic. METHODS/STUDY POPULATION: Point mutations on normal iPSCs was performed via Donorguide CRISPR/Cas9. The single stranded RNA/DNA donors contain genetic alterations of CHCHD10: Pro12Ser, Arg15Leu, Pro23Leu, Pro34Ser, Ser59Leu, Gly66Val, Pro80Leu, Tyr92Cys and Gln102His. Ribonucleoprotein electroporation was used to transfect iPSCs and DNA sequencing was used to validate gene editing. To validate transcriptional adaption, changes in levels of protein and gene expression were measured via immunoblot and quantification of CHCHD10 and CHCHCD2 was performed from whole cells lysates of the edited iPSCs. RESULTS/ANTICIPATED RESULTS: We anticipate that CHCHD2 transcriptional adaptation can functionally compensate for the locus loss of function of CHCHD10. This mechanism of transcriptional adaptation may contribute to an explanation for variation in clinical manifestations of patient phenotypes. DISCUSSION/SIGNIFICANCE: This study supplies further evidence for genetic modification as a treatment option for diseases with point mutation causal or enabling mechanisms, including some variants of ALS. Future work will explore the gene-correction from an ALS patient with a known CHCHD10-R15L variant.
323 Generation of a functional precision medicine pipeline which combines comparative transcriptomics and tumor organoid modeling to identify bespoke treatment strategies for glioblastoma
Authors: Reed Megan R. , Lyle A. Geoffrey , De Loose Annick , Learned Katrina , Vibat Cecile Rose T. , Wardell Christopher P. , Eoff Robert L. , Vaske Olena M. , Rodriguez Analiz
Published: April 2022

OBJECTIVES/GOALS: A functional precision medicine platform to identify therapeutic targets for a glioblastoma patient with Li Fraumeni syndrome was performed. Comparative transcriptomics identified druggable targets and patient derived organoids and a 3D-PREDICT drug screening assay was used to validate the pipeline and identify further therapeutic targets. METHODS/STUDY POPULATION: A comparative transcriptomics pipeline was used to identify druggable genes that are uniquely overexpressed in our patient of interest relative to a cancer compendium of 12,747 tumor RNA sequencing datasets including 200 GBMs. Mini-ring patient derived organoid-based drug viability assays were performed to validate the comparative transcriptomics data. Additionally, a spheroid-based drug screening assay (3D-PREDICT) was performed and used to identify further therapeutic targets. RESULTS/ANTICIPATED RESULTS: Using comparative transcriptomics STAT1 and STAT2 were found to be significantly overexpressed in our patient, indicating ruxolitinib, a Janus kinase 1 and 2 inhibitor, as a potential therapy. Druggable pathways predicted using comparative transcriptomics corresponded with ruxolitinib sensitivity in a panel of patient derived organoids screened with this compound. Cells from the LFS patient were among the most sensitive to ruxolitinib compared to patient-derived cells with lower STAT1 and STAT2 expression levels. Additionally, 3D-PREDICT screening identified the mTOR inhibitor everolimus as a potential candidate. These two targeted therapies were selected for our patient and resulted in radiographic disease stability. DISCUSSION/SIGNIFICANCE: This research illustrates the use of comparative transcriptomics to identify druggable pathways irrespective of actionable DNA mutations present. Our results are promising and serve to highlight the importance of functional precision medicine in tailoring treatment regimes to specific patients.
325 Blood pressure and the kidney cortex transcriptome response to high sodium diet challenge in female nonhuman primates
Authors: Riojas Angelica M. , Spradling-Reeves Kimberly D. , Shade Robert E. , Puppala Sobha R. , Christensen Clinton L. , Birnbaum Shifra , Glenn Jeremy P. , Li Cun , Shaltout Hossam , Hall-Ursone Shannan , Cox Laura A.
Published: April 2022

OBJECTIVES/GOALS: The goal of this study was to understand the impact of a high sodium diet on gene networks in the kidney that correlate with blood pressure in female primates, and translating findings to women. METHODS/STUDY POPULATION: Sodium-naÃ¯ve female baboons (n=7) were fed a low-sodium (LS) diet for 6 weeks followed by a high sodium (HS) diet for 6 weeks. Sodium intake, serum 17 beta-estradiol, and ultrasound-guided kidney biopsies for RNA-Seq were collected at the end of each diet. Blood pressure was continuously measured for 64-hour periods throughout the study by implantable telemetry devices. Weighted gene coexpression network analysis was performed on RNA-Seq data to identify transcripts correlated with blood pressure on each diet. Network analysis was performed on transcripts highly correlated with BP, and in silico findings were validated by immunohistochemistry of kidney tissues. RESULTS/ANTICIPATED RESULTS: On the LS diet, Na+ intake and serum 17 beta-estradiol concentration correlated with BP. Cell type composition of renal biopsies was consistent among all animals for both diets. Kidney transcriptomes differed by diet; analysis by unbiased weighted gene co-expression network analysis revealed modules of genes correlated with BP on the HS diet. Network analysis of module genes showed causal networks linking hormone receptors, proliferation and differentiation, methylation, hypoxia, insulin and lipid regulation, and inflammation as regulators underlying variation in BP on the HS diet. Our results show variation in BP correlated with novel kidney gene networks with master regulators PPARG and MYC in female baboons on a HS diet. DISCUSSION/SIGNIFICANCE: Previous studies in primates to identify molecular networks dysregulated by HS diet focused on males. Current clinical guidelines do not offer sex-specific treatment plans for sodium sensitive hypertension. This study leveraged variation in BP as a first step to identify correlated kidney regulatory gene networks in female primates after a HS diet.
340 Machine Learning Segmentation of Amyloid Load in Ligamentum Flavum Specimens From Spinal Stenosis Patients
Authors: Wang Andy Y. , Sharma Vaishnavi , Saini Harleen , Tingen Joseph N. , Flores Alexandra , Liu Diang , Safain Mina G. , Kryzanski James , McPhail Ellen D. , Arkun Knarik , Riesenburger Ron I.
Published: April 2022

OBJECTIVES/GOALS: Wild-type transthyretin amyloid (ATTRwt) deposits have been found to deposit in the ligamentum flavum (LF) of spinal stenosis patients prior to systemic and cardiac amyloidosis, and is implicated in LF hypertrophy. Currently, no precise method of quantifying amyloid deposits exists. Here, we present our machine learning quantification method. METHODS/STUDY POPULATION: Images of ligamentum flavum specimens stained with Congo red are obtained from spinal stenosis patients undergoing laminectomies and confirmed to be positive for ATTRwt. Amyloid deposits in these specimens are classified and quantified by TWS through training the algorithm via user-directed annotations on images of LF. TWS can also be automated through exposure to a set of training images with user- directed annotations, and then application to a set of new images without additional annotations. Additional methods of color thresholding and manual segmentation are also used on these images for comparison to TWS. RESULTS/ANTICIPATED RESULTS: We develop the use of TWS in images of LF and demonstrate its potential for automated quantification. TWS is strongly correlated with manual segmentation in the training set of images with user-directed annotations (R = 0.98; p = 0.0033) as well as in the application set of images where TWS was automated (R = 0.94; p = 0.016). Color thresholding was weakly correlated with manual segmentation in the training set of images (R = 0.78; p = 0.12) and in the application set of images (R = 0.65; p = 0.23). DISCUSSION/SIGNIFICANCE: Our machine learning method correlates with the gold standard comparator of manual segmentation and outperforms color thresholding. This novel machine learning quantification method is a precise, objective, accessible, high throughput, and powerful tool that will hopefully pave the way towards future research and clinical applications.
348 Resting state fMRI connectivity in individuals with aphasia and the role of the inferior frontal gyrus
Authors: Diedrichs Victoria , Osher David , Harnish Stacy
Published: April 2022

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.
351 Loop-mediated isothermal amplification for the detection of cytomegalovirus infection and drug resistance at the point of care
Authors: Medina-Rivera Melisa , Westblade Lars F. , Erickson David , Mehta Saurabh
Published: April 2022

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.
360 Retrospective Evaluation of Whole-Exome Sequencing in Puerto Ricans with Neurogenetic Complex Traits
Authors: Albino Elinette , Carlo Simon , Chapel-Crespo Cristel , Santiago-Cornier Alberto , Buxo Carmen
Published: April 2022

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.
373 Youth Nicotine Addiction: Strategic Defiance of Regulatory Oversight by the Disposable Electronic Nicotine Delivery System Industry
Authors: Riggs Griffin , Church Terry David
Published: April 2022

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.
374 Therapeutic Class Labeling of EGFR Companion Diagnostics (CDxs)
Authors: Chen Joyce , Cooper Lisa
Published: April 2022

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.
398 The Feasibility of Designing and Using a 3-D Printed Dynamic Upper Extremity Orthosis (DUEO) with Children with Cerebral Palsy and Severe Upper Extremity Involvement
Authors: Lee Justin J. , Shaikh Siraj Z. , Chu Alice
Published: April 2022

OBJECTIVES/GOALS: To evaluate whether a low cost, functional dynamic 3-D printed upper extremity (UE) orthosis could be designed, fabricated and used by children with cerebral palsy (CP) with severe unilateral UE involvement and the ability of common standardized instruments to assess its effectiveness. METHODS/STUDY POPULATION: Five patients, ages 13-17 years, with CP and unilateral UE involvement, MACS levels III-IV, were enrolled. Custom forearm thumb opponens orthosis and dynamic upper extremity orthosis (DUEO) were designed and fabricated by a multidisciplinary team for use during 8 one-hour occupational therapy sessions targeting bimanual UE training. Pre- and post-assessments included Assisting Hand Assessment(AHA), Melbourne-2(MA-2), Pediatric Motor Activity Log-Revised(PMAL-R), and PedsQL Measurement Model for the Pediatric Quality of Life Inventory: CP Module(Peds-QL). RESULTS/ANTICIPATED RESULTS: The 3D printed orthotic device is custom fit to the patient based on scans of their arm and is designed with a tensioning system that allows for functional usage of the arm. It incorporates a rigid polymer to provide support and flexible material for comfort where appropriate. Overall, higher post-treatment scores were found for the majority of participants using the custom orthotic. Four made minimal clinically important differences (MCID) in the AHA. Three met MCID scores in subtests of MA-2 (two positive and one negative). Patient-reported outcome improvements were found for PMAL-R for four, but only one met MCID and at least three reported improvements in more than one domain of PedsQL. DISCUSSION/SIGNIFICANCE: Children with CP may often present with UE impairment, yet current therapeutic orthoses only target the progression of contractures and are still limited by cost and discomfort. Our team designed and fabricated a functional, low cost, 3D printed orthosis that showed significant gains in UE function.
411 Essential role for the neurodevelopmental disorder-linked gene, MEF2C, in inhibitory neuron function and neurotypical behaviors
Authors: Cho Yongjoo (Jennifer) , Assali Ahlem , Tsvetkov Evgeny , Cowan Christopher W.
Published: April 2022

OBJECTIVES/GOALS: The MEF2 family of transcription factors regulate gene expression controlling cell differentiation and synapse development. Mutations or deletions in the MEF2C gene cause a neurodevelopmental disorder that includes symptoms of autism spectrum disorder. In this study, we aim to study the role of MEF2C in GABAergic populations using an animal model. METHODS/STUDY POPULATION: MEF2C Haploinsufficiency Syndrome (MCHS) occurs when there is one functional allele and one disrupted allele of MEF2C. To study the role of MEF2C in GABAergic populations during mouse development, we bred Vgat (vesicular GABA transporter)-Cre mice, which express cre recombinase broadly in early developing GABAergic neurons, with a floxed Mef2c loss-of-function mouse to create offspring that are GABAergic cell-specific Mef2c heterozygous mutants (Mef2c cHetVgat-cre). We then subjected these mutants and littermate controls to a battery of tests measuring MCHS-relevant phenotypes, including spatial working memory, anxiety-like behavior, social preference, sensory sensitivity, and Pavlovian learning and memory. RESULTS/ANTICIPATED RESULTS: Mef2c cHetVgat-cre mice showed significant deficits in spatial working memory, social preference, and contextual fear memory, all of which are prefrontal cortex (PFC)-dependent behaviors. Interestingly, we noted that conditional Mef2c knockout mice (Mef2c cKOVgat-cre) showed embryonic and early postnatal lethality, probable seizures, and severe motor coordination problems, highlighting the importance of MEF2C function in GABAergic populations. DISCUSSION/SIGNIFICANCE: We hypothesize that MEF2C plays a cell-autonomous role in GABAergic cells to control the balance of excitatory and inhibitory synaptic transmission in the developing and mature brain, which in the Mef2c cHet mice might be critical for PFC-dependent learning and memory and sociability.
415 Line1 Derepression in Specific Retrotransposon Families in Aged Mice Leads to Cytosolic DNA and Increased Inflammation
Authors: Martinez John , Simon Matthew , Seluanov Andrei , Gorbunova Vera
Published: April 2022

OBJECTIVES/GOALS: The major objective of this project is two points. First, is to repeat and confirm previous observations that there is elevated cytosolic Line1 DNA in the cytoplasm of cells derived from old mice compared to young. Second is to identify which Line1s in the genome are contributing to this free DNA and test if targeting them rescues the age-related phenotype. METHODS/STUDY POPULATION: This project will focus on data collected from both tissues and primary cells derived from multiple tissues. Using cellular/tissue fractionation kits, we isolate specifically from the cytoplasm. This specificity is confirmed by western blotting. Measurement of the Line1 levels is measured by quantitative PCR. Subsequently, these cytoplasmic samples are sent off for sequencing in order to quantify the length of the free DNA in the cytoplasm and to identify which Line1 genomic families the cytosolic DNA originates. Additionally, FISH is utilized to visualize Line1 DNA in the cytoplasm of aged versus young cells RESULTS/ANTICIPATED RESULTS: We anticipate this research to confirm the hypothesis that extranuclear Line1 DNA accumulates with age in both tissues and primary fibroblasts. Additionally, we expect to be able to determine which specific families of genomic Line1 is driving this extranuclear DNA, which would suggest the active retrotransopable elements that are directly involved in this aging related phenotype. Assuming successful identification of such families, we can then target and silence these specific elements to determine not only if cytoplasmic Line1 in aged mice decreases, but additionally if the healthspan and/or lifespan of these mice improves DISCUSSION/SIGNIFICANCE: Dereoressed Line1s have been shown to be involved in detrimental phenotypes, including autoimmune disease, cancer, and inflammaging. Targeting retrotransposons, either directly through degradation of transcriptional product of LINE1s or indirectly by improving function of regulators, will be crucial in ablating aging phenotypes
424 Considerations Mid-Translation of a Novel Extracellular Vesicle Product in Myocarditis
Authors: Beetler Danielle , Bruno Katelyn A. , Behfar Atta , Cooper Leslie T. , Fairweather DeLisa
Published: April 2022

OBJECTIVES/GOALS: Myocarditis is an inflammatory cardiomyopathy commonly caused by viral infections. The residual burden of this disease after guideline-based therapies is substantial, as there are no pathway-specific therapies. Our long-term goal is to find and translate treatments that reduce acute myocarditis severity and prevent progression of disease. METHODS/STUDY POPULATION: Of available therapies, extracellular vesicles (EVs) are ideally suited to the task of simultaneous, specific reprogramming of multiple pathogenesis mechanisms necessary in diseases like myocarditis, without similar immunosuppressive risks as their pharmacologic counterparts. We obtained plasma from healthy men and women and isolated nanoparticles, which were analyzed for physiochemical markers of human EVs. After confirming presence of EVs, we injected these plasma-derived extracellular vesicles (PEV) into male BALB/c mice vs. PBS control intraperitoneally on days -1, 0, 1 of viral infection (day 0) in a highly translational, mouse model of myocarditis. Hearts were examined at day 10 at the peak of acute myocarditis, using standard histological and cell composition analyses. RESULTS/ANTICIPATED RESULTS: Mice treated with PEV had significantly less myocardial inflammation both histologically and by gene expression of immune markers in the heart. The immunoregulation by PEV treatment decreased many key components of innate immune response networks that are known to be upregulated during acute myocarditis: TLR4+ mast cells and macrophages and complement. These pathways drive the profibrotic gene and protein changes that lead to remodeling, fibrosis, and disease progression observed in patients. We anticipate that when we analyze later timepoints in this model (day 35 post infection) which are normally associated with development of chronic myocarditis, dilated cardiomyopathy, we will see reduction of this fibrosis and of damage-associated changes. DISCUSSION/SIGNIFICANCE: These data suggest that EVs from plasma may be a novel treatment for viral myocarditis, but translation of EVs is hindered by their heterogeneity. We demonstrate characterization both physiochemically and functionally in a well-defined model. Such practices are necessary as these contemporary products challenge current regulation standards.
426 Long non-coding RNA WNT5A-AS1 shows sex-dimorphic differences in survival for males with glioblastoma
Authors: Miller Aunay , Wang Junwen , Loftus Joseph , Tran Nhan L
Published: April 2022

OBJECTIVES/GOALS: The goal of this study is to evaluate the role of WNT5A and WNT5a-AS1 in sex-differences of GBM progression. In our preliminary studies, we found that a long non-coding RNA WNT5A-AS1 is overexpressed in male GBM patients. We also found that WNT5A-AS1s expression shows a negative correlation with overall survival within male patients. METHODS/STUDY POPULATION: We will define the mechanism by which WNT5A-AS1 regulates WNT5a-mediated glioma stem cell (GSC) maintenance by assessing the effects of inhibiting WNT5A-AS1 expression on transcriptional activity and stemness in GSCs. We will determine if there are distinct Wnt-signaling patterns in male and female isogenic murine astrocytes by examining the expression of downstream proteins in the Wnt signaling pathway and how inhibition of WNT5A-AS1 alters this expression. We will then examine the impact of WNT5A-AS1 on temozolomide (TMZ) resistance in-vitro and in-vivo. We will assess the cell viability and survival of GBM PDX cells upon treatment with TMZ in vitro. Next, we will assess the capacity of knockdown of WNT5A-AS1 to increase sensitivity to TMZ-induced cell death and prolong survival in vivo in intracranial models. RESULTS/ANTICIPATED RESULTS: We hypothesize that WNT5A-AS1 targets Wnt5a and regulates its expression. We anticipate that knockdown of WNT5A-AS1 will upregulate WNT5A expression. We also expect that inhibiting WNT5A-AS1 will alter GSC stem maintenance and functional effects. We expect to see an increase in downstream Wnt5a signaling proteins in males vs females when treated with exogenous Wnt5a. We hypothesize that knockdown of both, WNT5A-AS1 and WNT5A will alter the expression of downstream proteins. We hypothesize that knockdown of WNT5A-AS1 will decrease tumor growth and therapeutic resistance to TMZ while increasing survival in patient derived xenographs in vivo and in vitro. DISCUSSION/SIGNIFICANCE: This study will provide insight into the mechanisms underlying the difference in GBM onset and progression between male and female patients, which is clinically important. We will also characterize the biological role WNT5A-AS1 which is currently unknown to date and elucidate differential role of GSCs in GBM progression between male and female.