OBJECTIVES/GOALS: Using the NIH’s expanded definition of underrepresented populations in the biomedical, clinical, behavioral and social science research enterprise, we examined the impediments for conducting translational research experienced by those from underrepresented groups. [https://acts.slayte.com/calls/detail/740a13de-316c-11ee-90f4-0e0ce905385c/draft/389221c1-434e-11ee-90f4-0e0ce905385c#_ftn1] #_ftn1 METHODS/STUDY POPULATION: One hundred and ninety-nine people completed a survey distributed to 750 persons who had interacted with our Center’s service cores as users, awardees, mentors, committee members, seminar attendees, and/or participated Center sponsored programming (response rate = 26.5%). The survey addressed barriers to conducting clinical and translational research at the respondent’s institution, awareness of and interest in using specific Advance RI-CTR services, and satisfaction with their institution’s efforts to support clinical and translational research. RESULTS/ANTICIPATED RESULTS: Women reported access to collaboration across institutions as a barrier to clinical and translational research that existed to a great extent (28%) significantly more than men (10%). More than half (53%) of the other underrepresented researchers surveyed identified insufficient grant administration supportas a barrier that occurs to a great extent, compared with 35% of researchers who were not from an underrepresented group. Other barriers reported more frequently among underrepresented researchers included lack of pilot project funding, inadequate space for conducting research, lower access to collaborators across institutions, and difficulty obtaining advice on regulatory issues and commercial development. DISCUSSION/SIGNIFICANCE: Efforts to address the barriers identified by underrepresented groups will include, but not be limited to, improving collaborations across institutions, support for grant administration, and a discussion of plans for the Center to augment and advocate at the partner institutions on behalf of these underrepresented individuals.

OBJECTIVES/SPECIFIC AIMS: To describe how Michigan Institute for Clinical & Health Research (MICHR) has engaged communities in its leadership and governance structure. This presentation will describe these practices, how they are being evaluated, and future plans for institute-wide engagement of communities in translational research. METHODS/STUDY POPULATION: Engaged partners from various communities across Michigan in various ways within MICHR’s Community Engagement Program. RESULTS/ANTICIPATED RESULTS: MICHR has utilized participatory practices in the development of the CAB to strengthen existing relationships and build new ones with potential partners. DISCUSSION/SIGNIFICANCE OF IMPACT: MICHR-wide Community Advisory Board (CAB) will ensure community voices are heard and utilized in leadership and strategic decisions for CTSA activities.

Objectives/Goals: To assess the attitudes toward social justice of residents and faculty following the didactic series and evaluate the impact of the longitudinal curriculum on resident engagement in social justice activities through community engagements. Methods/Study Population: Data were gathered through questionnaires collected from participating residents and faculty. The session topics were chosen based on “Precepting Toward Social Justice” curriculum from the University of Minnesota Department of Family Medicine and Community Health, including sessions on advocacy, diversity, and implicit bias, among others. The self-assessment questionnaires were specific to the topics of the lecture, assessing knowledge, skills, and attitudes before and after the education sessions. Participants were asked to rate these aptitudes of each topic from “none” to “expert.” Results/Anticipated Results: Out of 22 total participants, 16 returned surveys. Overall, there was a substantial improvement in aptitudes after the lecture. There was an increase in reported above average knowledge of topics: Agency (12.5% to 56.25%), Humility (18.75% to 62.5%), Dignity (12.5% to 62.5%), Partner (18.75% to 50%), Awaken (37.5% to 75%), Place (12.5% to 43.75%), Asset (0% to 37.5%), Sanctuary (6.25% to 50%), Belonging (0% to 37.5%), Whole (0% to 43.75%), Liberation (12.5% to 68.75%), and Heal (37.5% to 75%). 31% reported that the lectures alone had an above average influence on pursuing more social justice activities through community engagement. 44% reported that the lectures had an above average impact on their subsequent patient encounters. 69% stated that this series helped develop their overall social justice skills. Discussion/Significance of Impact: All patients deserve equitable care. Family physicians have a unique role in addressing social justice concerns to eliminate inequalities and drive for better health outcomes. By enforcing social justice in education and implementing it into practice, satisfaction, trust, and well-being of patient and provider will be improved.

OBJECTIVES/SPECIFIC AIMS: The specific objective of this proposal is to identify and validate targetable tumor-associated antigens (TAAs) in ovarian and pancreatic cancer. It is our central hypothesis that the accurate identification and selection of appropriate TAAs will provide a foundation on which to develop of novel and effective cancer immunotherapies. We have formulated this hypothesis on the basis of preliminary results in which we have used high-throughput tandem mass spectrometry (MS) to successfully identify TAAs from melanoma patient tumors. We have subsequently generated TAA-specific T-cells that showed specific recognition and killing of tumor cells, and will form the basis of an upcoming clinical trial for our melanoma patients. We now have extended this antigen identification pipeline into ovarian cancer to accomplish our objective of developing effective T-cell-based immunotherapies for ovarian cancer and pancreatic patients. METHODS/STUDY POPULATION: We have collected patient tumor specimens, and we performed HLA immunoprecipitation, peptide elution, and completed high-throughput tandem MS on these eluted samples to identify TAAs. In addition, we have validated the safety of potential targets through the use of the publicly available RNA sequence data sources GTEX and TCGA. RESULTS/ANTICIPATED RESULTS: To date, we have successfully completed over 60 peptide elutions from ovarian and pancreatic patients samples. In total, we have found several potential novel tumor-associated targets. VGLL1, is one of these identified antigens, and in conjunction with our collaborators, we have successfully generated T-cells against it. Additionally, we have found that VGLL1 is a potential novel TAA for 3 other cancer types, including bladder, gastric, and triple negative breast cancers. We are now focusing our efforts on testing these T-cells against additional ovarian cancer cell lines and these cancer types to determine their specificity. We plan to continue the generation and testing other identified potential TAAs as well. We plan to use these T-cells directly in clinical trials in the future. DISCUSSION/SIGNIFICANCE OF IMPACT: The rationale for this proposal is that through the identification and validation of TAAs, we can open the door to a new world of therapies that can potentially increase the survival rate in a disease with a historically grim prognosis.

Objectives/Goals: Undergraduate Medical Education (UME) may apply Just-in-Time training (JITT) to provide medical students with learning experiences closely aligned with real-time clinical needs. The purpose of this scoping review is to offer an overview of the implementation of JITT training in UME. Methods/Study Population: Following the five-stage framework by Arksey and O’Malley to methodically collect and analyze studies on JITT in UME, five electronic databases were searched, and a supplemental search for grey literature was conducted. Studies exploring the integration of JITT principles into UME clinical training and their time to follow-up after training were included. Bloom’s Taxonomy was used to assess educational goals of JITT interventions. Results/Anticipated Results: The review yielded 21 studies across 4 countries. The majority were cohort studies (13) and randomized control trials (5). Assessment definitions and use of JITT varied widely. Most studies focused on short-term outcomes, defined by being measured immediately after JITT session (15) or at the end of JITT-based rotation or clerkship (3). Three studies evaluated outcomes at a period longer than 2 weeks after completion of session or clerkship. Attitudes (9), followed by skills (8) were the most common educational goals of intervention. The efficacy and utility of JITT in improving educational goal acquisition was demonstrated in 90% (17/19) of the studies with reported outcomes. Discussion/Significance of Impact: The introduction of JITT in UME has been shown to meet the immediate needs of healthcare environments; however, evidence is limited in the evaluation of longer-term outcomes. Further research to determine the impact of JITT on long-term learning retention and education goal acquisition in UME is merited.

OBJECTIVES/GOALS: Over 50 million people in the U.S. have an eating disorder (EDs), and body dissatisfaction is a key precursor to EDs. The current study seeks to culturally adapt an evidence-based positive body image program administered through a mobile-phone application for use with Latinx women. METHODS/STUDY POPULATION: The positive body image program centers around body functionality, which in contrast to appearance-based body image, emphasizes the body’s capabilities, including physical capacities, health and internal processes, senses, creative endeavors, self-care, and communication with others. Latinx women aged 18-25 years-old (n =15) will be recruited to participate in focus groups and individual qualitative interviews. Interviews will examine how Latinx women relate to the concepts of positive body image and body functionality. Participants will also provide feedback on the application design and program content. RESULTS/ANTICIPATED RESULTS: Results will elucidate how Latinx women relate to their body and can be used to inform our understanding of Latinx women’s positive body image. Results will also inform what components of the intervention need to be tailored to be more relevant for use in this population. Information from the interviews will be used to explore adaptations to create a more acceptable and effective intervention prior to testing the efficacy of the program. DISCUSSION/SIGNIFICANCE: Most evidence-based body image programs are not developed with culturally sensitive programming in mind. Thus, culturally adapting an evidence-based positive body image program that could be widely disseminated through a mobile-application could address body image concerns in Latinx women.

OBJECTIVES/GOALS: Rising rates of youth obesity, diabetes, depression, and anxiety necessitate programs that address physical and mental health concurrently. We describe a feasibility study for DiscoverU, an afterschool mentoring program that integrates multiple aspects of health including social emotional learning, physical activity, and mindful eating. METHODS/STUDY POPULATION: Trained college students mentored middle and high school students in a Midwestern school district. DiscoverU was delivered 2 hours, 2 days/week for 8 weeks. Based on self-determination theory, DiscoverU was designed to meet National Afterschool Association healthy eating and physical activity and social emotional learning standards. We assessed feasibility with participant attendance (middle, high school, and college students) and acceptability through qualitative data from participants and relevant stakeholders regarding facilitators/barriers to program implementation. We observed indicators of mentoring, lesson fidelity, and assessed physical activity using accelerometry. Pre-post surveys measured self-realization, self-regulation, mindful eating, and physical activity self-efficacy. RESULTS/ANTICIPATED RESULTS: We expect DiscoverU to be feasible and well accepted. We anticipate attendance will be similar or higher than that of other afterschool programs in the district. From focus groups we expect to hear a variety of factors that facilitate/prevent program engagement and learn about the acceptability of specific lessons. We expect to gain insight on processes and procedures from school stakeholders that will inform the sustainability of DiscoverU. We expect program fidelity to be high and mentoring skills to improve over the course of the program. We anticipate the majority of participants will meet National Afterschool Alliance physical activity guidelines. Preliminary outcomes of self-determination, self-regulation, mindful eating, and physical activity self-efficacy are expected to improve over the program. DISCUSSION/SIGNIFICANCE: Findings will help determine the readiness of DiscoverU to be scaled to other schools. A subsequent randomized effectiveness study will evaluate DiscoverU’s impact on intervention mechanisms (e.g., self-determination, self-efficacy) as well as on physical activity, diet, weight, and depression/anxiety symptomology.

OBJECTIVES/SPECIFIC AIMS: To use a systematic and iterative process to develop and refine toolkits to support dissemination and implementation (D&I) research. METHODS/STUDY POPULATION: Participants included research staff from the Dissemination and Implementation Research Core (DIRC), a research methods core from the Institute of Clinical and Translational Science at Washington University in St. Louis, other D&I experts from the University, and national experts from the D&I field. This project used education design research methodology and a systematic and iterative process involving several phases. The first phase (preliminary research and initial development) consisted of analysis of the educational problem and its context, and led to the development of toolkit prototypes and plans for their implementation. In the second phase (development and formative evaluation), toolkits were iteratively evaluated with emphasis on content validity and consistency and effectiveness as perceived by the users. Finally, in the summative evaluation, the toolkits were evaluated based on their use as intended. RESULTS/ANTICIPATED RESULTS: Our team identified the target audience as DIRC customers and investigators from disciplines across the University, and found that resources for beginners to D&I were lacking. The team developed 8 toolkits: (1) Introduction to D&I; (2) How to develop D&I Aims; (3) D&I Designs; (4) Implementation Outcomes; (5) Implementation Organizational Measures; (6) Assessing Barriers and Facilitators; (7) D&I Designs; and (8) Guideline research. These prototypes were iteratively revised for content validity and consistency. Finally, each toolkit was evaluated by two national experts in D&I science, and further refined. DISCUSSION/SIGNIFICANCE OF IMPACT: This systematic and cyclical process led to the development of 8 toolkits to support researchers in D&I science, which are now available on the DIRC Web site. This set the stage for development of new toolkits as additional needs are identified.

ABSTRACT IMPACT: As technologies emerge at an increasing pace, the product developed through this work will guide rural health experts through a repeatable method of technology evaluation and selection at a faster and more reliable pace than otherwise possible. OBJECTIVES/GOALS: New technologies are emerging at an increasing pace, which leads to the question: ‘how is one to select a specific technology for their research?’ In response, this project endeavored to develop a technology evaluation and selection framework for rural health researchers. METHODS/STUDY POPULATION: The approach selected for this project included three phases. Phase one was to gain an understanding of rural health challenges, health-related emerging technologies, and rural health resources. Phase two involved using the information from phase one to select and adapt a set of technology foresight and forecasting analysis tools to be compiled within a framework. The third phase of the project was to prototype the framework, obtain researcher feedback, and iteratively implement improvements. Recommendations for the future of the framework were also developed during the third phase. RESULTS/ANTICIPATED RESULTS: The resulting product is the ‘Rural Health: Evaluation and Selection of Technology (RHEST) Framework.’ The RHEST Framework is a guide made available to use to aid in technology selection during the development of a new rural health project. The framework guides researchers through various stages, including ideation, analysis, and decision. Technology analysis tools are introduced in each stage, with links to additional information. The guide also contains a resource catalog for quick information look-up to find data sources, funding opportunities, and expert connections. Quantitative and qualitative data captured indicate that the product would add value for rural health researchers. DISCUSSION/SIGNIFICANCE OF FINDINGS: The initial version of the RHEST framework is limited in value because it is a static document and the primary audience are researchers. The value potential could improve considerably, however, if the framework were expanded to be a dynamic resource available to rural health care providers.

OBJECTIVES/SPECIFIC AIMS: The Institute for Transnational Sciences (ITS) has developed novel methods to ethically engage stakeholders across the transnational research spectrum, up to and including public health practice and policy. METHODS/STUDY POPULATION: In 2014, the ITS co-founded The Research, Education, And Community Health (REACH), the mission of which was to facilitate communication, collaborative research, and service activities between faculty and scientists and area community leaders. The intent was to identify and meet the needs of our communities without gaps and/or redundancies, thus better leveraging time, funding, and efforts. RESULTS/ANTICIPATED RESULTS: REACH now boasts 23 Centers, Departments, and Institutes, as well as 39 community organizations, including public and mental health agencies, clinicians, policy makers, family service centers, cultural and faith-based organizations, business, and local schools/colleges. We offer 3 methods for consideration as best practices: (1) a comprehensive community health needs assessment, (2) an “Offer and Ask” community/campus partnership mechanism, and (3) Community Science Workshops, based on the European Union’s Science Shops. DISCUSSION/SIGNIFICANCE OF IMPACT: Results of REACH’s work have been used to provide guidance for enhanced, data-driven programs and allocation of resources for local and statewide initiatives. The organization has evolved into an independent coalition seeking 501(c)3 status and is planning to expand its scope to 5 counties. REACH thus serves as model for successful replication across applicable CTSA hubs.

OBJECTIVES/GOALS: In 2021, Frontiers CTSI revamped its evaluation infrastructure to be comprehensive, efficient, and transparent in demonstrating outputs and outcomes. We sought to build a platform to standardize measures across program areas, integrate continuous improvement processes into operations, and reduce the data entry burden for investigators. METHODS/STUDY POPULATION: To identify useful metrics, we facilitated each Core’s creation of a logic model, in which they identified all planned activities, expected outputs, and anticipated outcomes for the 5-year cycle and beyond. We identified appropriate metrics based on the logic models and aligned metrics across programs against extant administrative data. We then built a data collection and evaluation platform within REDCap to capture user requests, staff completion of requests, and, ultimately, request outcomes. We built a similar system to track events, attendance, and outcomes. Aligning with other hubs, we also transitioned to a membership model. Membership serves as the backbone of the evaluation platform and allows us to tailor communication, capture demographic information, and reduce the data entry burden for members. RESULTS/ANTICIPATED RESULTS: The Frontiers Evaluation Platform consists of 9 redcap projects with distinct functions and uses throughout the Institute. Point-of-service collection forms include the Consultation Request Event Tracking. Annual Forms include a Study Outcome, Impact, and Member Assessment Survey. Set timepoint collections include K & T application, Mock Study Section, and Pilot grant application submission, review, and outcomes. Flight Tracker is used to collect scientific outcomes and integrated with the platform. Using SQL, the membership module has been integrated into all forms to check and collect membership before service access and provide relevant member data to navigators. All relevant data is then synched into a dashboard for program leadership and management to track outputs and outcomes in real-time. DISCUSSION/SIGNIFICANCE: Since the launch of the evaluation platform in Fall 2022, Frontiers has increased its workflow efficiency and streamlined continuous improvement communication. The platform can serve as a template for other hubs to build efficient processes to create comprehensive and transparent evaluation plans.

OBJECTIVES/GOALS: AA are over-represented on the waitlist for kidney transplant and are often unaware of how waitlist acceptance practices differ across transplant programs and influence access to transplant. We will develop a culturally sensitive transplant program report card to communicate these variations. METHODS/STUDY POPULATION: Scientific Registry of Transplant Recipients (SRTR) data will be used to identity clinical factors strongly associated with AA access to transplant. Interviews and focus groups with AA kidney transplant candidates and their families will collect feedback on the SRTR report card and inform the development of the culturally sensitive report card. Additional focus groups will evaluate its effect on knowledge and medical decision making. We will collaborate with the stakeholders, including AA transplant candidates and their families, transplant programs, SRTR, and providers, to identify strategies to disseminate the report card in the AA community RESULTS/ANTICIPATED RESULTS: To date, no investigation has systematically collected feedback on the SRTR transplant program report card from AA candidates to ensure that the tool is accessible and effective in the AA community. We hypothesize that a culturally sensitive report card will improve AA candidates’ knowledge of program factors that impact access to transplant and enable informed decisions about where they pursue a transplant evaluation. The results of this study have the potential to change how AA patients are counselled while seeking transplantation. DISCUSSION/SIGNIFICANCE OF IMPACT: A culturally sensitive report card can reach more AA patients and enable more informed decision making by providing education about differences in transplant programs that may impact their access to transplant. In the future, we will design a trial to evaluate the prototype.

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.

OBJECTIVES/SPECIFIC AIMS: More men than women develop urinary stones and their prevalence alters in women with menopause suggesting a steroidal influence. In men the incidence of stones is highest during July and August suggesting that environmental factors such as Vitamin D (VitD), a steroid, may affect stone formation. Previous studies have found differences in the development of stones between men and women; however, the reasons for sex differences in stone formation and type remain unclear. METHODS/STUDY POPULATION: We examined VitD levels in men and women (n = 18,753) that had no diseases based on a lack of an ICD-9 or ICD-10 code in their electronic medical record. We found that normal, healthy women had significantly higher levels of sera VitD compared to men (p = 6x10-6). We then examined whether sex differences existed for key endpoints/data from the Mayo Clinic Urinary Stone Disease (USD) Registry, which has around 1,600 urinary stone patients that are well-phenotyped according to sex, age and stone type. RESULTS/ANTICIPATED RESULTS: Control women were found to have higher sera VitD levels than men, but the sex difference no longer exists in kidney stone disease patients. When we further separated by race, we found that differences in VitD levels reappeared; this suggests that race also plays a role in sera VitD variances. DISCUSSION/SIGNIFICANCE OF IMPACT: We are developing a disease severity score, which we will use to correlate to sera VitD levels in patients according to sex, age and race. Future analyses will take into account whether subjects had VitD and calcium supplementation. This project begins to explore the mechanism behind the sex differences known to exist in urinary stone disease, which is critically needed to provide improved diagnosis and therapy for this debilitating disease.

OBJECTIVES/SPECIFIC AIMS: Discuss ethical and policy issues that will impact clinical research. Raise awareness of the need to understand internal policies at home institutions Encourage further examination of ways to facilitate clinical research participation. METHODS/STUDY POPULATION: Ethical and policy analysis. RESULTS/ANTICIPATED RESULTS: Ideally, clinical research participants should not be required to pay to participate in research. However, if we go with an equity model, as opposed to an equality model, policies should be changed to allow equal access to research participation. This is a matter of justice and also will enhance the quality of the science. DISCUSSION/SIGNIFICANCE OF IMPACT: Unless steps are taken to make participation in clinical research less burdensome financially for participants, research may slow or results may be biased, because only those who can pay will be able to participate.

OBJECTIVES/GOALS: Aims 1&2: Develop (1) and implement (2) online, guided self-help intervention for ED psychopathology and weight reduction. Aim 3: Follow-up to track remission of ED psychopathology and symptoms and WL maintenance at end of treatment and 6-months. METHODS/STUDY POPULATION: N = 60 college students meeting criteria (clinical/sub-clinical binge-type ED with BMI > 25) will complete a baseline survey and then will be randomized into a condition. Students in the intervention group (n = 30) will be offered 8 weeks of an online, guided self-help intervention for ED and WL. Students in the control group (n = 30) will receive an email message to seek support from Student Health Services. All participants will receive follow-ups 9 weeks and 6 months after baseline. Data analysis will compare Eating Disorder Examination Questionnaire (EDE-Q) scores and WL (change in BMI) at all three time-points. Group comparisons will be assessed via two-way mixed-model ANOVA. RESULTS/ANTICIPATED RESULTS: Recruitment is still ongoing. Data collected by the time of the conference will be presented on the poster. DISCUSSION/SIGNIFICANCE OF IMPACT: Online, guided self-help interventions have been used for WL, as well as for treatment of EDs separately, but no program exists to manage these commonly comorbid conditions concurrently. Thus, this pilot study will examine the effectiveness of combined programs to breach this treatment gap.

OBJECTIVES/GOALS: Recent research has attempted to identify diagnostic, prognostic, and predictive biomarkers, however, currently, no biomarkers can accurately diagnose GBC and predict patients prognosis. Using machine learning, we can utilize high-throughput RNA sequencing with clinicopathologic data to develop a predictive tool for GBC prognosis. METHODS/STUDY POPULATION: Current predictive models for GBC outcomes often utilize clinical data only. We aim to build a superior algorithm to predict overall survival in GBC patients with advanced disease, using machine learning approaches to prioritize biomarkers for GBC prognosis. We have identified over 80 fresh frozen GBC tissue samples from Rochester, Minnesota, Daegu, Korea, Vilnius, Lithuania, and Calgary, Canada. We will perform next-generation RNA sequencing on these tissue samples. The patients clinical, pathologic and survival data will be abstracted from the medical record. Random forests, support vector machines, and gradient boosting machines will be applied to train the data. Standard 5-fold cross validation will be used to assess performance of each ML algorithm. RESULTS/ANTICIPATED RESULTS: Our preliminary analysis of next generation RNA sequencing from 18 GBC tissue samples identified recurrent mutations in genes enriched in pathways in cytoskeletal signaling, cell organization, cell movement, extracellular matrix interaction, growth, and proliferation. The top three most significantly altered pathways, actin cytoskeleton signaling, hepatic fibrosis/hepatic stellate cell activation, and epithelial adherens junction signaling, emphasized a molecular metastatic and invasive fingerprint in our patient cohort. This molecular fingerprint is consistent with the previous knowledge of the highly metastatic nature of gallbladder tumors and is also manifested physiologically in the patient cohort. DISCUSSION/SIGNIFICANCE: Integrative analysis of molecular and clinical characterization of GBC has not been fully established, and minimal improvement has been made to the survival of these patients. If overall survival can be better predicted, we can gain a greater understanding of key biomarkers driving the tumor phenotype.

OBJECTIVES/SPECIFIC AIMS: To develop a KL2 curriculum on the science and art of drug formulation. METHODS/STUDY POPULATION: Develop training materials for KL2 scholars that outline the art of formulation development. Materials will include syllabi, reading materials, and course work. RESULTS/ANTICIPATED RESULTS: This will enhance the training of KL2 scholars by incorporating formulation development concepts into their human health enhancing research projects. DISCUSSION/SIGNIFICANCE OF IMPACT: For new chemical entities, formulation goals must be realistic and move along in a step-wise manner from the laboratory bench, through toxicology studies, and on to Phase 1 studies. By training scholars in phase-specific formulation goals, their interactions with funding agencies, formulation scientists, and regulators will be more efficient, productive, and successful. For those scholars who are working to improve existing treatments, introducing the concept of formulation improvements that can create new indications, or improve efficacy, safety and patient compliance will open up more possibilities for creative product development.

OBJECTIVES/GOALS: Rib fractures are common traumatic thoracic injuries and are associated with high rates of morbidity and mortality. In those age ≥ 65, the rate of these complications double. This study sought to identify the extent to which injury-related predictors influence clinical outcomes in geriatric patients with rib fractures. METHODS/STUDY POPULATION: A retrospective 5-year review was performed of a single Level 1 Trauma center registry. Geriatric patients (≥65 years of age) diagnosed with rib fractures from January 1, 2014 to December 31, 2019 were included. The primary outcome of interest was in-hospital mortality. Secondary outcomes included hospital and intensive care unit length of stay (HLOS and ICU LOS, respectively) and discharge disposition, as a surrogate for loss of independence. Further, subgroup analysis based on number of rib fractures (i.e. <4 and ≥4 rib fractures) was performed. RESULTS/ANTICIPATED RESULTS: 2,134 adult trauma patients were admitted with at least one rib fracture. Of these, 1,037 (49%) were ≥ 65 years old. This cohort had a mean age of 78.6 years old, injury severity score (ISS) of 11.4, HLOS of 7.4 days and 29% required ICU care with mean ICU LOS of 1.9 days. Only 36% were discharged home compared to 64% who were discharged to a care facility and thus had a loss of independence. Overall mortality was 6.3%. Compared to survivors, non-survivors had a higher ISS (19.3 vs. 10.8, p = < 0.0001) and longer ICU LOS (7.1 vs. 6.5 days, p = 0.04). Analysis based on number of rib fractures showed that those with ≥4 rib fractures had significantly higher mortality (8% vs. 4%, p = 0.008), longer HLOS (8.7 vs. 6.1 days, p < 0.0001), longer ICU LOS (2.6 vs. 1.3 days, p < 0.0001), and significantly lower discharge to home (32% vs. 39%, p = 0.02). DISCUSSION/SIGNIFICANCE: To our knowledge, this is the largest single-center study of geriatric patients with rib fractures. In this study, the observed mortality in patients ≥65 years of age was 6.3% which represents a lower mortality rate than historically reported. Despite this, only 36% were able to be discharged directly to home.

OBJECTIVES/SPECIFIC AIMS: The off-target and organ-specific toxicities observed in cancer immunotherapy present an obstacle to T-cell-based therapeutics. A recent clinical trial underscored the need for improved methods to define TCR specificity after melanoma patients treated with TCR engineered T-cells suffered from fatal cardiovascular toxicity arising from the unpredicted recognition of a muscle-specific peptide. METHODS/STUDY POPULATION: To address this drawback to T-cell-based immunotherapies, we developed a novel protein engineering approach to define the peptide specificity of a given TCR. Here, directed evolution in a yeast display system produced a large scale peptide library, where recognition by the melanoma reactive DMF5 TCR acted as the guiding selective pressure. After this technique identified a panel of putative cross reactive peptides, sequence analysis and computational modeling followed by kinetic binding experiments and structural analysis determined the DMF5 TCR recognizes 2 distinct classes of peptides through chemically distinct mechanisms. RESULTS/ANTICIPATED RESULTS: This information led to the rational, structure-based design of additional cross reactive peptides and introduced a unique approach to screen the human proteome and identify the TCR targets which triggered undesired autoimmunity when this molecule was used in clinical trials. DISCUSSION/SIGNIFICANCE OF IMPACT: The distinct chemical nature of the 2 peptide classes suggest TCRs are more cross reactive than previously thought, presenting an obstacle to cell-based immunotherapy. Defining the peptide specificity of TCRs is of high interest to the immunology community, and will lead to improved approaches to designing engineered TCRs for cell therapy.