OBJECTIVES/GOALS: Understanding the mechanism of transcriptional adaptation may contribute to an explanation for variation in clinical manifestations of Amyotrophic lateral sclerosis patient phenotypes. METHODS/STUDY POPULATION: To examine transcriptional adaptation, we utilized gene editing tools in HT1080 cells and patient samples with known CHCHD10 mutations causative for Amyotrophic lateral sclerosis. Frameshift mutations were performed via CRISPR-Cas9. Ribonucleoprotein electroporation was used to transfect cells and DNA sequencing was conducted to validate gene editing. To validate transcriptional adaption, changes in levels of protein and gene expression will be measured via immunoblot and quantification of CHCHD10 and CHCHCD2 from whole cells lysates of the edited cells. 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: Our approach would advance discovery science towards by exploring CHCHD10/2 transcriptional adaptation mechanism that can lead to novel therapies for rare Amyotrophic lateral sclerosis, such as CHCHD10-R15L.

The COVID-19 pandemic accelerated the development of decentralized clinical trials (DCT). DCT’s are an important and pragmatic method for assessing health outcomes yet comprise only a minority of clinical trials, and few published methodologies exist. In this report, we detail the operational components of COVID-OUT, a decentralized, multicenter, quadruple-blinded, randomized trial that rapidly delivered study drugs nation-wide. The trial examined three medications (metformin, ivermectin, and fluvoxamine) as outpatient treatment of SARS-CoV-2 for their effectiveness in preventing severe or long COVID-19. Decentralized strategies included HIPAA-compliant electronic screening and consenting, prepacking investigational product to accelerate delivery after randomization, and remotely confirming participant-reported outcomes. Of the 1417 individuals with the intention-to-treat sample, the remote nature of the study caused an additional 94 participants to not take any doses of study drug. Therefore, 1323 participants were in the modified intention-to-treat sample, which was the a priori primary study sample. Only 1.4% of participants were lost to follow-up. Decentralized strategies facilitated the successful completion of the COVID-OUT trial without any in-person contact by expediting intervention delivery, expanding trial access geographically, limiting contagion exposure, and making it easy for participants to complete follow-up visits. Remotely completed consent and follow-up facilitated enrollment.

OBJECTIVES/GOALS: This study explored whether gabapentin (GBN) differentially impacted heart rate variability (HRV) and whether HRV was associated with opioid withdrawal ratings among participants with opioid use disorder (OUD) undergoing a randomized, double blind placebo-controlled, trial (RCT) of GBN during a buprenorphine (BUP)-assisted taper. METHODS/STUDY POPULATION: Participants (ages 18-64) with OUD, no recent use of benzodiazepines/barbiturates, and no major psychiatric disorders or unstable medical conditions were enrolled in the RCT, inducted onto BUP starting week 1 day 1 and randomly assigned to receive adjunct GBN or placebo starting week 1 day 3. All participants began a 10-day BUP-taper beginning week 2 day 3. HRV measures were assessed on week 1 day 2 (before GBN/placebo induction), week 2 day 2, and week 3 day 5 (end of BUP taper). HRV metrics were analyzed using Two Sample T-Test to determine differences between GBN vs. Placebo. Correlations between HRV metrics and opioid withdrawal ratings administered at the above timepoints will be analyzed using Spearman correlation. RESULTS/ANTICIPATED RESULTS: 28 participants underwent at least 1 HRV session that resulted in usable data. Preliminary statistical analyses revealed that HRV trended lower for GBN subjects during PB exercises than Placebo subjects, demonstrated by a higher mean heart rate for GBN subjects compared to Placebo subjects (p=0.0506) at the end of the BUP-taper (week 3 day 5). We expect future analyses to demonstrate a negative correlation between certain HRV metrics indicative of parasympathetic tone and opioid withdrawal rating assessment scores indicative of withdrawal severity. Such findings would demonstrate an association between opioid withdrawal severity and lower parasympathetic tone and HRV. DISCUSSION/SIGNIFICANCE: Individuals with OUD have previously been shown to have a lower parasympathetic tone than individuals without OUD. Additionally, opioid withdrawal has been shown to be associated with reduced parasympathetic tone. Our initial findings suggest that adjunct GBN administration was not associated with lower parasympathetic tone during PB exercises.

OBJECTIVES/GOALS: A quarter of Puerto Rican pediatric population are overweight or obese. Pediatric obesity is established as a global public health concern that will develop liver complication as a Non-alcoholic fatty liver disease (NAFLD). This study aims to determine the association between body weight, liver size and texture in Puerto Rican pediatric population. METHODS/STUDY POPULATION: Twenty-nine (n=29) pediatric participants (20 unhealthy weight and 9 healthy weight) between 7 to 19 years underwent panoramic ultrasound imaging of the RLL. Craniocaudal RLL measurement and liver texture were evaluated. Body mass index and waist circumference were also compared. Shapiro-Wilk test and students t-tests were attained with significance at p < 0.05. RESULTS/ANTICIPATED RESULTS: Statistical differences were detected between healthy weight and unhealthy weight pediatric patients: (1) RLL craniocaudal diameter (p=0.006); (2) waist circumference (p < 0.001); (3) BMI (p < 0.001). Unhealthy weight (overweight and obese) pediatric patients presented a greater number of livers with fat infiltration (13) and hepatomegaly (15). DISCUSSION/SIGNIFICANCE: This preliminary results showed that liver size was larger and fatty infiltration are relatively frequent among overweight and obese pediatric patients. Prevention of unhealthy weight and the diagnosis of changes in liver texture and size among pediatric patients is important to prevent progressing of liver disease and avoid irreversible damage.

In 2016, Duke reconfigured its clinical research job descriptions and workforce to be competency-based, modeled around the Joint Taskforce for Clinical Trial Competency framework. To ensure consistency in job classification amongst new hires in the clinical research workforce, Duke subsequently implemented a Title Picker tool. The tool compares the research unit’s description of job responsibility needs against those standardized job descriptions used to map incumbents in 2016. Duke worked with human resources and evaluated the impact on their process as well as on the broader community of staff who hire clinical research professionals. Implementation of the tool has enabled Duke to create consistent job classifications for its workforce and better understand who composes the clinical research professional workforce. This tool has provided valuable workforce metrics, such as attrition, hiring, etc., and strengthened our collaboration with Human Resources.

OBJECTIVES/GOALS: Glioblastoma (GBM) is the most common and aggressive adult primary brain malignancy. Clinically, GBM is refractory to T cell immune checkpoint blockade (ICB), in part due to its dense immune suppressive myeloid stroma. Here we show that myeloid-targeting STING agonists can repolarize the GBM microenvironment to cure ICB-refractory GBM models. METHODS/STUDY POPULATION: Using the synthetic cyclic di-nucleotide STING agonist IACS-8803 (8803) we treated orthotopic ICB-refractory QPP8 orthotopic murine GBM tumors intratumorally. We then analyzed survival and performed high parameter flow cytometry profiling of the tumor immune microenvironment following STING agonist treatment. To assess the contribution of adaptive immunity to STING agonist therapeutic efficacy, we treated orthotopic QPP8 tumors implanted in RAG1 KO mice and monitored survival. RESULTS/ANTICIPATED RESULTS: We found that STING agonist therapy cured murine orthotopic QPP8 tumors, in contrast to ICB that showed no survival benefit. In RAG1-/- mice bearing QPP8 tumors STING agonist therapy extended survival, however, the curative effect observed in wild-type mice was lost in the absence of adaptive immunity. STING agonist-treated QPP8 tumors displayed increased counts of CD8 T cells and NK cells, and decreased CD8 T cell PD1 expression. Infiltration of STING-treated gliomas by Ly6C+ F4/80+ Mono-MDSC substantially increased; however, these cells expressed reduced CD206 and CD163, suggestive of reduced immuno-suppression. Finally, in the cervical LN of QPP8-treated mice the frequency and CD80/CD86 expression of cDC1 cells increased.​ DISCUSSION/SIGNIFICANCE: ICB has failed in GBM, and the suppressive myeloid stroma remains a major barrier to generating anti-GBM T cell responses. Our work shows that STING activation, which primarily targets innate immunity myeloid cells 'upstream' of T cells in the antitumor immunity cycle, can cure ICB-refractory GBM tumors in an adaptive immunity-dependent manner.

OBJECTIVES/GOALS: To describe cognitive and psychiatric symptom profiles of individuals with bvFTD and hyperorality. We test two hypotheses: (1) individuals with hyperorality show more severe psychiatric profiles and (2) neuroanatomic correlates of hyperorality in advanced bvFTD differ from those with early bvFTD. METHODS/STUDY POPULATION: Participants were enrolled in ALLFTD–a multi-site longitudinal study in FTD. We selected the 354 participants who had a primary clinical diagnosis of bvFTD, 344 of whom had data on hyperorality. Each participant underwent extensive clinical interviews and examinations, structural neuroimaging, and blood sampling. Five anatomic regions of interest were identified and analyzed based on previously identified neuroanatomic correlates of hyperorality. Differences in participant characteristics and clinical outcomes were compared using t-tests for continuous variables and Pearsonχ2 tests for categorical variables. Linear multivariate regression controlling for age and total intracranial volume (TIV) was used to examine associations between atrophy in regions of interest and hyperorality status. RESULTS/ANTICIPATED RESULTS: Early-stage participants with hyperorality had poorer self-monitoring, empathic concern, and perspective taking as well as higher CDR behavioral subscale scores compared to those without hyperorality. Advanced stage participants with hyperorality had higher scores on the Social Behavior Observer Checklist compared to those without hyperorality. Early-stage participants with hyperorality displayed higher rates of ritualistic/compulsive behavior and motor disturbance. Advanced stage participants had higher rates of apathy, ritualistic/compulsive behavior, anxiety, and elation. In the advanced stage participants, hyperorality was associated with atrophy in the right dorsal striatum, the right ventral striatum, and the right insula cortex. DISCUSSION/SIGNIFICANCE: Hyperorality emerges early and is accompanied by neuropsychiatric symptoms prior to significant neurodegeneration. Overtime, participants with hyperorality develop more psychiatric symptoms as well as atrophy in striatal and insular brain regions. Our findings suggest a role for novel interventions like non-invasive brain stimulation.

OBJECTIVES/GOALS: Microtubule poisons, like Taxol, are used to treat triple negative breast cancer (TNBC) and may induce lethal aneuploidy in cancer cells. Patients initially respond, but often develop drug resistance. New targeted drugs that cause aneuploidy may be a valuable approach to therapy. One potential target is the Kinesin 13 MCAK, which limits aneuploidy. METHODS/STUDY POPULATION: TCGA and GSE47561 databases were probed for MCAK expression, and data was stratified by subtype and survival statistics. Knockdown studies were performed to test whether MCAK knockdown sensitizes cells to taxanes for cell proliferation and for induction of aneuploidy. FRET and image-based screens were used to identify MCAK inhibitors from small molecule inhibitor libraries. Inhibitors were then tested for functional effects in multiple cell-based assays and for clonal growth in colony formation assays. RESULTS/ANTICIPATED RESULTS: MCAK expression is upregulated in TNBC and associated with reduced overall survival. Knockdown of MCAK caused a two-to-five-fold reduction of the IC50 for Taxol in cancer cell lines, with no change in normal cell lines. Taxol treatment or MCAK knockdown increased aneuploidy induction, with no additive effect between the two. Our small molecule screen identified three putative MCAK inhibitors, which induced aneuploidy in both taxane-sensitive and taxane-resistant cells. These inhibitors also reduced clonogenic growth, and the most potent inhibitor, C4, caused an approximate five-fold reduction in the IC50 for Taxol in cell proliferation assays. DISCUSSION/SIGNIFICANCE: MCAK can serve as a biomarker of breast cancer prognosis. MCAK knockdown or inhibition sensitizes cancer cells to Taxol without affecting normal cells, making it a potential target in combination therapy. MCAK inhibitors also reduce growth as single agents in taxane resistant lines, giving them potential use as therapies in resistant disease.

Clinical and translational research relies on a well-trained workforce, but mentorship programs designed expressly for this workforce are lacking. This paper presents the development of a mentoring program for research staff and identifies key programmatic outcomes. Research staff participating in this program were matched with a senior mentor. Focus groups were conducted to identify key program outcomes. Surveys were administered throughout the program period to assess participants’ experience, gains in skill, and subsequent careers. Analysis of the resultant qualitative and quantitative data are used to characterize the implementation and impact of the program. A total of 47 mentees and 30 mentors participated in program between 2018 and 2023. A comprehensive logic model of short-, intermediate- and long-term outcomes was developed. Participants reported positive valuations of every programmatic outcome assessed including their program experience, learning and research careers. The pool of available mentors also grew as new mentors were successfully recruited for each cohort. This mentorship program developed and implemented by senior research staff successfully provided junior research staff with professional development support, mentorship, and professional development opportunities. Junior and senior health research staff built mentoring relationships that advanced their clinical and translational research careers.

OBJECTIVES/GOALS: Our objective is to establish a proteomic protein labeling method from tumor tissue and blood samples obtained from patients undergoing surgery for pancreatic cancer in Puerto Rico. Our goal is to discover potential biomarkers in the patient tumor/blood samples that are not expressed in normal control samples obtained from potential organ donors. METHODS/STUDY POPULATION: A pilot study with ten patients undergoing surgery for pancreatic cancer will obtain tumor tissue and blood samples. Protein extracts isolated from tissue/cells will be reduced, alkylated, and digested overnight. Samples will be labeled with TMT reagents and mixed before fractionation and cleanup. Labeled samples will be analyzed with a high-resolution Orbitrap LC-MS/MS before data analysis to identify peptides and quantify the reporter ions. The altered proteins will be analyzed by ELISA to confirm their presence. The protein arrangements will be compared with results from proteomic profile banks to assess their prevalence. As controls, parallel protein analyses will be performed on normal tissue/blood samples from organ donors, facilitated by our local organ procurement organization. RESULTS/ANTICIPATED RESULTS: We anticipate finding proteogenomic material defining PC and new proteomic subtypes not previously described in this population. In addition, studying protein overexpression and underexpression can identify relevant genes and potential biomarkers. We hypothesize that PC in the Hispanic population will show slight variations in tumor protein expression than in other populations, which could lead to the discovery of a new Hispanic-specific biomarker. DISCUSSION/SIGNIFICANCE: We expect to provide essential information that will influence the next steps in developing future screening tests. Identifying specific proteins with the potential to become a preventive test should eventually lead to a reduction in morbidity and mortality of PC. The results of this work should lay the foundation that can guide future research.

In 2016, the National Center for Advancing Translational Science launched the Trial Innovation Network (TIN) to address barriers to efficient and informative multicenter trials. The TIN provides a national platform, working in partnership with 60+ Clinical and Translational Science Award (CTSA) hubs across the country to support the design and conduct of successful multicenter trials. A dedicated Hub Liaison Team (HLT) was established within each CTSA to facilitate connection between the hubs and the newly launched Trial and Recruitment Innovation Centers. Each HLT serves as an expert intermediary, connecting CTSA Hub investigators with TIN support, and connecting TIN research teams with potential multicenter trial site investigators. The cross-consortium Liaison Team network was developed during the first TIN funding cycle, and it is now a mature national network at the cutting edge of team science in clinical and translational research. The CTSA-based HLT structures and the external network structure have been developed in collaborative and iterative ways, with methods for shared learning and continuous process improvement. In this paper, we review the structure, function, and development of the Liaison Team network, discuss lessons learned during the first TIN funding cycle, and outline a path toward further network maturity.

This paper outlines the development, deployment and use, and testing of a tool for measuring and improving healthcare researcher embeddedness – i.e., being connected to and engaged with key leverage points and stakeholders in a health system. Despite the widely acknowledged importance of embeddedness for learning health systems and late-stage translational research, we were not aware of useful tools for addressing and improving embeddedness in scholar training programs. We developed the MN-LHS Embeddedness Tool covering connections to committees, working groups, leadership, and other points of contact across four domains: patients and caregivers; local practice (e.g., operations and workflows); local institutional research (e.g., research committees and agenda- or initiative-setting groups); and national (strategic connections within professional groups, conferences, etc.). We used qualitative patterns and narrative findings from 11 learning health system training program scholars to explore variation in scholar trajectories and the embeddedness tool’s usefulness in scholar professional development. Tool characteristics showed moderate evidence of construct validity; secondarily, we found significant differences in embeddedness, as a score, from baseline through program completion. The tool has demonstrated simple, practical utility in making embeddedness an explicit (rather than hidden) part of applied and learning health system researcher training, alongside emerging evidence for validity.

OBJECTIVES/GOALS: Fecal incontinence is associated with increased caregiver strain, reduced patient dignity and diminished quality of life. A spectrum of incontinence exists, along with a paucity of available solutions for patients and their caregivers. This research aims to stratify this space and identify gaps within the existing solution landscape. METHODS/STUDY POPULATION: To understand this problem, a literature review was performed with key search terms specific to fecal incontinence. These included, anal incontinence epidemiology, fecal incontinence in nursing homes, and incontinence management. To determine gaps within the existing solution landscape, key search terms related to existing solutions for fecal incontinence were also included. These included, fecal management systems, rectal incontinence therapies, and anorectal incontinence procedures.' To perform a population segmentation, white papers, review articles, and cross-sectional studies were reviewed to break down the burden of incontinence in older adults living in nursing facilities and in the community. RESULTS/ANTICIPATED RESULTS: Two unaddressed populations were identified, the first being independent adults over the age of forty, particularly women, who suffer from frequent, bothersome incontinence. These 1.2 million patients are active, living at home, and they restrict their daily activities due to incontinence. However, there are several durable and effective solutions for patients who have sufficient sphincter tone or who are surgical candidates. The second population identified are caregiver dependent older adults residing in nursing facilities who suffer from severe incontinence. This population of 160,000 is affected more severely by consequences of fecal and are poorly served by solutions that are largely absorptive such as diapers and pads. DISCUSSION/SIGNIFICANCE: Although two populations were identified, caregiver dependent older adults residing in nursing homes were identified to have a significant unmet need within incontinence care. Current solutions are onerous and transient, preventing ease and duration for use by caregivers and nurses.

Improving the quality and conduct of multi-center clinical trials is essential to the generation of generalizable knowledge about the safety and efficacy of healthcare treatments. Despite significant effort and expense, many clinical trials are unsuccessful. The National Center for Advancing Translational Science launched the Trial Innovation Network to address critical roadblocks in multi-center trials by leveraging existing infrastructure and developing operational innovations. We provide an overview of the roadblocks that led to opportunities for operational innovation, our work to develop, define, and map innovations across the network, and how we implemented and disseminated mature innovations.

OBJECTIVES/GOALS: Diabetes is a group of chronic metabolic diseases and significant gaps remain in our understanding of disease etiology, treatment regimens, and diabetes-related complications. The objective of study is to demonstrate how informatics techniques can leverage real-world data for diabetes research and identify barriers for implementation. METHODS/STUDY POPULATION: We evaluated informatics applications of real-world data in diabetes research conducted by the Facelli Research Group. The types of real-world data were categorized into clinical records, diabetes-related repositories, wearable sensors, and other data sources. Translational informatics applications were characterized into thematic groups of 1.) use of electronic health records, registries, and claims and other data sources to generate real-world evidence, 2.) evolution of novel methods to accelerate generation and use of real-world data, and 3.) infrastructure to support the generation and use of real-world data in translational science. A literature review is being conducted to identify additional articles meeting these themes focused on diabetes research. RESULTS/ANTICIPATED RESULTS: 6 research projects were included for analysis. The diabetes-focus spanned type 1 diabetes, type 2 diabetes, and general diabetes mellitus. Informatics methods included machine learning and data mining while real-world data sources included electronic medical records, the Environmental Determinants of Diabetes in the Young (TEDDY) study, continuous glucose monitors, and the U.S. Environmental Protection Agency (EPA) air pollution monitors. Overall, computability of real-world data, linkage of medical concepts to standardized terminologies, volume of data, and adoption of novel artificial intelligence methods were major determinants of successful implementation. Future work will systematically evaluate informatics applications of real-world data in diabetes from the academic community at large. DISCUSSION/SIGNIFICANCE: Translational informatics approaches are poised to leverage real-world data and better understand diabetes etiology, treatment regimens, and diabetes-related complications. By understanding barriers and opportunities for informatics methods, we can expedite translational applications in diabetes research.