Objectives/Goals: Increasingly, women and people of color are earning PhDs. However, historically underrepresented (HU) degree seekers often follow a “nontraditional” track, pursuing their degree part time, at an older age, and balancing work and family with their studies. Over half of the U.S. PhD students are part time, a feature correlated with candidate diversity. Methods/Study Population: To address this need, the Advancing Diversity and Equity in Pre-doctoral Trainees (ADEPT) program was developed for part-time HU students who do not meet the requirements of the NIH-funded NRSA TL1 program but who offer relevant work and life experience. The program provides individualized navigation and tailored support based on individual student need and career goals, flexible training opportunities, and intentional connections to established, dedicated, and well-trained mentors throughout the predoctoral journey. To better understand their unique needs, we conducted thematic analysis on the notes taken during monthly meetings to explore the challenges our students face and how ADEPT provides support in navigating these challenges. Results/Anticipated Results: Students pursuing a PhD on a part-time basis while balancing other responsibilities, such as caretaking or a full-time job, bring a unique perspective to their graduate education. 5 ADEPT students described how their background or clinical work experience played a crucial role in their decision to pursue a PhD. Themes from the monthly meeting and mentor meeting notes include the challenges of balancing full-time jobs with school, and how they do not experience the cohort effect that a traditional, full-time PhD student would have as a resource. The students reflected on how an individualized culturally sensitive approach was instrumental to their success. Discussion/Significance of Impact: It was initially believed that part-time PhD students were rare. Despite this misconception, we easily recruited students. Students reported their experiences do not reflect the experiences of their full-time peers and their difficulty establishing a peer group, demonstrating the clear need for ADEPT.

OBJECTIVES/GOALS: Sexual minority men (SMM) of color are at greater dementia risk than their White and non-SMM counterparts. The purpose of this study is to examine the relative effects of ethnoracial minority status, sexual minority status, and the intersection of both on dementia risk for men in the U.S. using subjective cognitive decline (SCD) as a proxy for dementia. METHODS/STUDY POPULATION: Data will be weighed to represent population estimates from 35 states’ 2019-2022 Behavioral Risk Factor Surveillance System Survey to describe SCD among 4 study groups consisting of SMM and non-SMM who are from minoritized ethnoracial groups (i.e., minoritized ethnoracial gay and bisexual men, minoritized ethnoracial heterosexual men), and White SMM and White non-SMM (i.e., White gay and bisexual men, White heterosexual men) aged ≥45 years. Logistic regressions and adjusted odds ratios will determine SCD prevalences and test for differences between prevalence and demographic characteristics (i.e., education, employment, income, marital status, health coverage, and depression). RESULTS/ANTICIPATED RESULTS: Due to excess chronic stress from belonging to multiple intersecting marginalized identities, we expect that SMM from minoritized ethnoracial groups will have a greater prevalence and higher odds of SCD after accounting for demographic factors when compared to groups that benefit from both “White Privilege” and heterosexism (White non-SMM, White SMM, and minoritized ethnoracial non-SMM). We also expect the odds of SCD to be higher in minoritized ethnoracial SMM when compared to White non-SMM and minoritized ethnoracial non-SMM. The odds of SCD are expected to be higher in White SMM compared to White non-SMM. DISCUSSION/SIGNIFICANCE: This is the first population-based study to apply an intersectional lens to examine dementia risk across the axes of race, sexual orientation, and gender identity. Findings from this study can inform targeted interventions to address the needs and experiences of LGBTQ+ subgroups at risk or living with dementia.

OBJECTIVES/GOALS: The project goal was to conduct formative work using community-based participatory research (CBPR) to inform the development of a laundromat-based reproductive health literacy intervention (LI)informed by Passhe-Orlow and Wolf’s Conceptual Model. We provide insights to support the development of similar health literacy interventions. METHODS/STUDY POPULATION: In Phase I, we recruited female patrons over the age of 18 living in DC from 4 community laundromats. Participants completed a survey with validated measures to assess reproductive health knowledge, self-efficacy, and intentions to seek care. Open-ended questions assessed participants’ experiences accessing care, communication with health providers, and laundromat use. Data informed the development of a 2-3 minute reproductive health literacy intervention (LI) storyboard. In Phase II, in-depth interviews were conducted with reproductive health providers (midwives, obstetricians-gynecologists, women’s health registered nurses, women’s health nurse practitioners) working in the communities where the laundromats are located. LI acceptability and appropriateness were explored. RESULTS/ANTICIPATED RESULTS: For Phase I, we recruited 60 patrons. Participants (68.3%) expressed interest in a laundromat-based reproductive health literacy intervention (LI). Identified reproductive knowledge gaps included: unaware of fertile days (50.0%); believe prenatal care should begin after 2nd trimester of pregnancy (50.0%); unclear about pregnancy spacing (85.0%); unable to identify use for folic acid (91.7%); believe once a cesarean always a cesarean (53.3%). In Phase II, all 14 providers supported increasing community accessibility to health information: “we have to move beyond doing public health education in a provider’s office. That’s very outdated.” Providers identified the LI as an approachable, informative, and accessible way to disseminate health information. DISCUSSION/SIGNIFICANCE: Patron and provider input informed our choice of content, style, and delivery of the reproductive health literacy intervention. An essential aspect of our CBPR approach, this research will inform effective, appropriate reproductive health education despite barriers presented by access, transportation, and work schedules.

Objectives/Goals: The Clinical and Translational Research (CTR) Pathway aims to increase the number of health science professionals across the WWAMI Region (Washington, Wyoming, Alaska, Montana, and Idaho) with significant awareness, interest, and experience in research. Methods/Study Population: Two cohorts of health sciences students have completed the formal coursework of the CTR Pathway, including sixty-three medical students from across the WWAMI region. Cohorts were organized into an Alaska in-person group, a Seattle in-person group, and a regional virtual group. Coursework includes a year-long series of three elective classes addressing key concepts of designing, conducting, and presenting results of a research project. Students engage in a dedicated full time 8-week summer research experience under the guidance of a research mentor. The culminating curriculum focuses on strong scientific communication by preparing students for poster and oral presentations at regional and national conferences. Results/Anticipated Results: The ultimate goal of the CTR pathway is to increase the number of health providers across the WWAMI region participating in CTR during their educational experience and subsequent professional careers. Many health sciences students hesitate to engage in research due to a perceived lack of necessary skills. The CTR pathway addresses this gap by equipping students with the research competencies needed to participate confidently in scientific inquiry. Feedback from the first two cohorts has been overwhelmingly positive, with many students highlighting how the CTR pathway enhanced their confidence and knowledge, empowering them to execute their proposed research projects and presentations successfully. The third cohort of students joined the CTR pathway in January 2026. Discussion/Significance of Impact: The CTR pathway positions early health sciences students to engage in research more deeply during their education and prepares them to seek additional training opportunities toward a career in research. Additionally, students located at regional sites and have a strong interest in practicing in rural areas.

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.

OBJECTIVES/SPECIFIC AIMS: Since 1971, Naloxone has been the only FDA approved opioid antagonist indicated for use after opioid overdose. New formulations of Naloxone have been introduced into the market, including an injectable, auto-injector, and nasal spray. However, Naloxone is short-acting and as such often requires multiple doses and may induce severe withdrawal symptoms. This study examines the regulatory framework to understand the evolution of products indicated to treat opioid overdose and the landscape of therapies in development. Furthermore, this study examines how the Food and Drug Administration (FDA) and other government agencies have approached the opioid crisis. METHODS/STUDY POPULATION: A PubMed search of “naloxone AND opioid overdose” with the filter “humans” was conducted to understand Naloxone’s regulatory framework. The term “naloxone” was searched on the Drugs@FDA: Approved Drug Products database. Additionally, “nalmefene” was searched on ClinicalTrials.gov. To examine the opioid antagonist market landscape, a PubMed search of “opioid antagonist AND opioid overdose” with the filters “humans” and “clinical trial,” and a ClinicalTrials.gov search of “opioid antagonist and opioid overdose,” were conducted. Government agency reports were reviewed and cataloged. RESULTS/ANTICIPATED RESULTS: Preliminary findings suggest a lack of innovation in the development of novel opioid antagonists. Most literature review findings focused on already-marketed Naloxone products, including the original injectable approved in 1971, the 2014 Evzio Auto-Injector, and the 2015 Narcan Nasal Spray (Figure 1). For example, there were 14 results yielded from the FDA approvals database, but none of these results represented a new opioid antagonist molecule. A longer-acting opioid antagonist, Nalmefene injectable, was approved in 1995 but has since been removed from the market due to low sales. Our initial ClinicalTrials.gov search using condition “opioid overdose” and other terms “opioid antagonist”,revealed no new studies being conducted on alternative opioid antagonist treatments for opioid overdose. Findings only focused on the distribution, co-dispensing, intervention, pharmacokinetics/pharmacodynamics (PK/PD) of Naloxone (Figure 2). However, a Google search yielded one new trial with an opioid antagonist by Opiant Pharmaceuticals, almost fifty years after FDA’s approval of Naloxone. A ClinicalTrials.gov search was then performed using the search term “nalmefene” to find whether Opiant Pharmaceuticals’ trial was in the ClinicalTrials.gov database. However, the Opiant trial is phase I, and as such does not require reporting on ClinicalTrials.gov. In 2017, the National Institutes of Health (NIH) launched an initiative for longer-acting opioid antagonist formulations. In 2018, Opiant Pharmaceuticals announced positive phase I results for intranasal Nalmefene. The potential return of Nalmefene in intranasal form may play a significant role in reducing overdoses, especially in cases where a longer-acting opioid antagonist is necessary. Opiant Pharmaceuticals’ trial commenced after the NIH announced their initiative; furthermore, the NIH’s National Institute on Drug Abuse granted the company $7.4 million to further the investigation of this drug. We will continue to research drugs that have previously been studied for the indication of treating opioid overdose in the United States and abroad and catalog them. DISCUSSION/SIGNIFICANCE OF IMPACT: The abuse and misuse of opioids in the United States has caused an epidemic accounting for over 115 opioid-overdose deaths each day, devastating our nation, both socially and economically. The United States spends $78.5 billion annually to combat the misuse of these drugs. Due to the severity of the opioid crisis, efforts to better understand approved therapies and investigational products in development to treat opioid overdose will be of significance moving forward. This research can inform agencies who are developing strategies to reduce opioid overdoses and pharmaceutical product developers about the current opioid antagonist landscape.

Objectives/Goals: To assess participation, satisfaction, and scholarly outcomes of IMPRS, a structured 10-week summer research program for rising second-year medical students at Indiana University School of Medicine (IUSM), one of the largest medical schools in the nation, across six cohorts (2018-2023). Methods/Study Population: We conducted a retrospective analysis of 1,090 IMPRS participants across six cohorts (2018–2023) at IUSM’s nine regional campuses. Students engaged in clinical, translational, community-based, and health outcomes research, culminating in an annual poster symposium. Program data included application numbers, participation rates, and satisfaction surveys. Scholarly outcomes were assessed via systematic PubMed and Scopus searches for publications derived from student projects. Search strategies combined student and mentor names, with results matched against symposium abstracts based on shared title keywords, content, topics, findings, or methodology. These matches were then used to calculate publication rates, time to publication, and analyze trends across cohorts. Results/Anticipated Results: Application rates increased from 23% (2017) to 76% (2023), with 21% to 61% ultimately participating. From 2017 to 2023, applications rose 77% (155 to 274), and participants increased 114% (103 to 220). Over six years, 1,090 students across IUSM’s nine campuses took part in IMPRS, resulting in 313 unique publications. The overall publication rate was 30.83%, with a mean time to publication of 22.6 months (IQR 13.0–30.8). Participant satisfaction remained high, with over 90% of students agreeing the program was a positive and productive experience. Discussion/Significance of Impact: IMPRS demonstrates a strong model for structured summer research, with a 30.83% publication rate comparable to similar programs (24-34%). High satisfaction and sustained growth indicate IMPRS successfully fosters scholarly development and cultivates future physician-scientists.

OBJECTIVES/SPECIFIC AIMS: Opinions regarding translational science vary incredibly. We aimed to gather a baseline of perceptions, barriers, and needs for translational science among faculty investigators. We will use these data to define areas in which the Duke Multidisciplinary Education and Research in Translational Science program (MERITS) can work to address, educate and improve. METHODS/STUDY POPULATION: Data was collected via a scalar, multiple-choice, open-ended survey including questions regarding, definition, impact, barriers, resources, and training preferences specific to translational science. Digital survey links were emailed to Duke University faculty. RESULTS/ANTICIPATED RESULTS: In total, 350 responses were collected. While perceptions of translational science varied, common defining elements were noted, including multidisciplinary collaboration (69%) and transitions between research stages (63%). Translational science was said to have an overall positive impact, despite 37% of participants stating issues of insufficient institution-wide support and 62% citing minimal training in translational science skills. DISCUSSION/SIGNIFICANCE OF IMPACT: Effective support for translational science requires a multi-faceted approach, as perceptions differ among investigators and between career stages. Duke MERITS will seek to standardize education and support ranging from teambuilding to entrepreneurship, and to promote support from institutional leadership to reduce barriers and facilitate acceleration of translational science.

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-naïve 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.

OBJECTIVES/SPECIFIC AIMS: Unlike the high cure rates (90%) of children with acute lymphoblastic leukemia (ALL), that of adults is still lagging behind and better therapies are needed. Maternal embryonic leucine-zipper kinase (MELK) is aberrantly upregulated in cancer, and implicated in cancer stem cell survival. A recent study has identified FOXM1, a MELK substrate, as a therapeutic target in B cell ALL (B-ALL). Thus, we hypothesized that MELK may act as a therapeutic target in ALL via targeting FOXM1 activity. METHODS/STUDY POPULATION: Western blot and qPCR were used to assess MELK expression in 14 ALL cell lines. Knock-down and kinase inhibition approaches targeting MELK expression and function, followed by CCK-8 and Annexin V (flow cytometry) assays to measure cell viability and apoptosis, respectively. RESULTS/ANTICIPATED RESULTS: MELK was significantly upregulated in patients with ALL (oncomine data analysis). MELK was also significantly higher in B-ALL and T-ALL cell lines compared with that in blood cells of healthy donors. MELK knock-down significantly decreased cell viability (40%–70%, p<0.05, Fig. 1) in ALL cells, and induced apoptosis (~40%). OTS167, a potent MELK inhibitor exhibited cytotoxic activities in both B and T-ALL cells. The IC50 of OTS167 ranged from 20 to 60 nM; we also found a significant increase in apoptosis (p<0.05). Mechanistically, MELK inhibition resulted in decrease of FOXM1 protein levels 3 hours post-treatment. DISCUSSION/SIGNIFICANCE OF IMPACT: MELK is highly expressed in ALL and represents a novel therapeutic target likely via modulating FOXM1 activity. Functional and mechanistic studies will complement and ensure the success of the undergoing Phase I/II clinical trial of OTS167 in patients with refractory or relapsed AML, ALL, and other advanced hematologic malignancies.

ABSTRACT IMPACT: This presentation highlights an integrated curriculum in CTR and a scientific entrepreneurship approach to entice and support students and faculty in HP programs into CTR and SE thus expanding the pool of new minority CTR researchers. OBJECTIVES/GOALS: To present the TVMSC as a hub for trainings, mentoring programs, courses, entrepreneurship and support activities for health professionals(HP) and HP students :graduate (GS) and UgS and UgF. Responding to the need for CTR minority researchers, in a virtual setting due to COVID-19 crisis. METHODS/STUDY POPULATION: TVMSC will offer an educational program based in the Center for Research,Entrepreneurship and Scientific Collaboration (CRESCO) with on line courses and workshops in CTR and SE, for HP and students and a continued education curriculum for HP and clinician scientists toward a certification in CTR. Two hands-on experiences: a) a Pilot project program(PiP) with teams composed of an F, that previously completed training cycles and a research experience from a previous project in CTR as PI, with a research mentor and students or an established researcher as a PI with UgS and UgF, and b) participation in a SE team which will engage in training and submission of an SE project proposal. RESULTS/ANTICIPATED RESULTS: By the end of the five-year period the project will have had 200 UgS, 200 GS and 200 F that received online assistance in CTR skills, statistics and SE; 48 UgS and 48 GS with the skills in SEFL. In curricular development the project expects to have 6 online tutorials created, one FLSE online course and 18 modules in CTR content areas available for continued education of HP. Certifications in CTR will be completed by 160F/HPs. The expected participation in CTR on-hands experiences is 32 F, 64 students and 32 established researchers. PiP teams will publish at least 8 scientific papers and SEFL teams will submit at least 5 SE project proposals and 100% increase in CRESCO web based resources DISCUSSION/SIGNIFICANCE OF FINDINGS: This Project and its expected results will provide students and faculty members island-wide with the knowledge, skills and experiences in CTR with IE approach to foster the expansion of a cadre of Hispanic minority CTR researchers in direct benefit of the health of the people of Puerto Rico.

Social determinants of health affect clinical and translational research processes and outcomes but remain underreported in empirical studies. This scoping review examined the rate and types of social determinants of health (SDoH) variables included in the JCTS translational research studies published between 2017 and 2023 and included 129 studies. Most papers (91.7%) reported at least one SDoH variable with age, race and ethnicity, and sex included most often. Future studies to inform the role of SDoH data in translational research and science are recommended, and a draft SDoH data checklist is provided.

Museum engagement may be an effective approach for decreasing social disconnection and pain among individuals living with chronic pain. In October 2019, we launched a randomized controlled trial to assess the feasibility of museum engagement for individuals living with chronic pain; the study was halted in March, 2020 due to Covid-19-related safety concerns. This paper describes the process of transitioning from in-person to virtual museum programing in order to continue the study. Virtual museum programing is a feasible option for individuals living with chronic pain that is amenable to research and which may improve accessibility, inclusivity, and scalability relative to in-person programing.

In the spring of 2020, New York City was at the epicenter of the COVID-19 pandemic in the USA, resulting in disruption of TL1 and KL2-mentored Clinical and Translational Science (CTS) research at the Icahn School of Medicine at Mount Sinai (ISMMS). The impact of the pandemic on trainees’ research productivity and career plans was explored using a qualitative survey. Participant responses were analyzed using coding and categorization. Six key themes emerged: redirection of effort, reduced access to people, lack of access to resources, home as a workplace, future uncertainty, and stress and anxiety. Insight into participant experiences allows for the development of support strategies and resources to address trainee needs.

OBJECTIVES/SPECIFIC AIMS: The Hispanic Clinical and Translational Education and Career Development program entails formal research training (Phase I) through an established post-doctoral Master of Science in Clinical and Translational Research. The most qualified graduates from Phase I compete to receive 1–2 years support for continued mentoring and career development (Phase II program) aiming to apply for a regular research grant or career award (K or R series). OBJECTIVE: This project aims to present an evaluation of the Phase II program and Scholars outcomes. METHODS/STUDY POPULATION: METHODS: Participants (n=12) responded to a semistructured interview including 43 questions about program’s processes and outcomes. Descriptive and content analysis was done. RESULTS/ANTICIPATED RESULTS: RESULTS: Results show that 83% are women, 42% are MD, and 67% are affiliated to the University of Puerto Rico-Medical Sciences Campus and 67% were able to fulfill their career development expectations during the Phase II Award. At present (92%) are conducting clinical research in their current position. Outcomes include new selection of research line, K Awards, and enhanced skills in clinical and translational research DISCUSSION/SIGNIFICANCE OF IMPACT: DISCUSSION: Challenges identified were: time management, better coaching and a more structured mentoring experience. The main benefit of the program were protected time, research budget, and the opportunity to acquire more research experience.

OBJECTIVES/GOALS: The Institute of Translational Health Science (ITHS)Remote Technologies for Research Reference Center (REMOTECH) aims to ease the use of remote technologies in research, addressing barriers to research participation related to in-person and onsite assessments METHODS/STUDY POPULATION: We engaged with stakeholders in our CTSA catchment area to understand the use and impact of remote research: implementation practices, participant acceptability, and impact on accrual. This environmental scan consisted of three parts: (1) Facilitated discussion groups with clinical trial regulatory and implementation experts regarding 10 key areas of research operations using composite case studies of research using remote technologies; (2) Semi-structured interviews with research teams who have successfully implemented remote technologies, querying about specific technologies, expected and actual barriers, and impact on the study population; and (3) A survey distributed to 900+ faculty and staff identifying prevalence of hybrid or fully remote research and describing specific remote technologies. RESULTS/ANTICIPATED RESULTS: Discussion group preliminary findings show experts recommended prioritizing the assessment of value and burden for both research participants and the research team, that equity and diversity should not be sacrificed to accommodate cost and efficiency, and the importance of evaluating the impact of implementing remote technologies on data collection and analysis. Seventeen of 30 interviews are complete, mid-point analysis shows researchers wanting formal best practices and training in remote research, desire increasing diversity through remote options, and expressing concerns about participant burden. 46% of survey respondents report implementation barriers, including participant burden and confusing regulatory pathways. 17% thought remote technologies were not appropriate for their studies. DISCUSSION/SIGNIFICANCE: We plan to leverage a multidisciplinary team to address the identified barriers and disseminate through a public remote technology information portal. Coding and further analysis is underway, including additional interviews targeting researchers working with adolescents and older adults with an increased focus on equity and diversity.

OBJECTIVES/SPECIFIC AIMS: The current treatment for amyotrophic lateral sclerosis (ALS) includes systemic delivery of neurotrophic factors (NTFs). Although this approach may seem theoretically sound, NTF efficacy within the central nervous system (CNS) is largely limited by the blood-brain barrier. Thus, a cell-based approach, which allows for targeted delivery of molecular therapies locally from the CNS, could lead to a paradigm shift in the field. METHODS/STUDY POPULATION: The Windebank and Staff group at Mayo Clinic completed a Phase I dose-escalation safety trial of autologous, adipose-derived mesenchymal stem cells (adMSCs) in an effort to move toward personalized medical treatment of ALS. The adMSCs were injected into the intrathecal space by lumbar puncture in 27 patients and the results showed an excellent safety profile across a range of doses. The team is moving forward with this idea by using gene-editing technology to develop clinical-grade, genetically modified autologous MSCs. The patient-derived adMSCs are modified at defined “safe-harbor” regions of the human genome through transcription activator-like effector nuclease (TALEN) technology. RESULTS/ANTICIPATED RESULTS: Our results show that electroporating adMSCs with plasmid DNA leads to efficient GFP or TALEN transgene expression, but yields low cell survival and a low rate of genetic modification. DISCUSSION/SIGNIFICANCE OF IMPACT: It can be concluded that: (1) TALEN technology may be used to target safe harbor loci for gene integration to produce therapeutic adMSC for ALS. (2) Primary barriers to adMSC modification are inefficient TALEN and donor template uptake, low cutting efficiency, and poor cell survival after electroporation. Future directions include optimizing the protocol to obtain 48 base pairs in the homology arms and increasing transfection efficiency.