OBJECTIVES/GOALS: Maternal opioid use disorder (OUD) is linked to poor fetal outcomes. While it has been established that opioids can cross from maternal to fetal circulation, the mechanisms underlying these adverse outcomes remain poorly defined. This study aims to uncover OUD-associated immunological changes in maternal and fetal circulation. METHODS/STUDY POPULATION: To study the effect of maternal OUD on the maternal immune system at delivery, we collected maternal blood samples at delivery from healthy pregnant women (controls) and pregnant women with a diagnosis of severe OUD. To study the impact of maternal OUD on newborn immunity, we also collected umbilical cord blood (UCB) at delivery. Flow cytometry was used to determine the frequency and phenotype of circulating immune cell subsets and responses to stimulation. Isolated monocytes were stimulated with bacterial/viral agonist cocktails, while T and NK cells were stimulated with PMA/ionomycin. The impact of maternal OUD on circulating immune mediators was determined by Luminex and on monocyte activation markers sCD14 and CRP by ELISA. RESULTS/ANTICIPATED RESULTS: In maternal circulation, OUD was associated with a significant decrease in markers of inflammation, cell proliferation, and activation. Frequencies of immune cell subsets were impacted by OUD, shown by an expansion of CD8+ EMRA T cells, marginal-zone B cells, mDCs, non-classical monocytes, and CD16low NK cells. While no differences were seen in T and NK cell responses to stimulation, monocytes and pDCs had significantly lower responses to bacterial and viral agonist stimulation. Analysis of UCB revealed increased levels of pro-apoptotic/T cell exhaustion mediators and pro-inflammatory cytokines, albeit decreased levels of several chemokines and growth factors. The UCB immune landscape is altered with maternal OUD, as demonstrated by a shift from naive to memory CD8+ T cells and a decrease in pDC frequency. DISCUSSION/SIGNIFICANCE: OUD dampens maternal peripheral immunity, possibly contributing to poor placental function or premature/delayed labor. Monocytes and pDCs lack antimicrobial functionality, suggesting increased infection susceptibility with OUD. Finally, these implications extend to the fetal compartment, shown by heightened immune activation in UCB.

OBJECTIVES/GOALS: Renal fibrosis is a critical pathophysiological event in chronic kidney diseases. Our goal is to determine the ability of dual-inhibitor of transforming growth factor beta receptor 1 (TGFα²R1) and mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4), TK850, on reducing kidney fibrosis. METHODS/STUDY POPULATION: To test the renal anti-fibrotic action ofdual TK850,8-10-week-old male and female C57BL/6mice with unilateral ureteral obstruction (UUO) induced kidney fibrosis were used. Mice were separated into 3 groups: group 1 contained mice that had UUO surgery (UUO control), group 2 contained mice prophylactically treated with TK850 thatstarted 7 days prior to UUO(UUO-P,20 mpk/d/ip), and group 3 contained mice interventionally treated with TK850 that started3 days after UUO(UUO-I,20 mpk/d/ip). Ten days following UUO the kidneys and blood were collected for analysis. Renal fibrosis was assessed from hydroxyproline content (measure of collagen)and histological collagen analysis using Picrosirius red stain. RESULTS/ANTICIPATED RESULTS: Renal hydroxyproline was increased equally in the UUO kidney of male (5.4 ± 0.41 µg/10mg, n=5) and female mice (5.5 ± 0.50 µg/10mg, n=5) compared to the contralateral control kidney (2.9 ± 0.14 µg/10mg, n=10). TK850 treatment in UUO-P mice (n=10, 3.4 ± 0.24 µg/10mg) and UUO-I mice (4.30 ± 0.20 µg/10mg, n=10) had significantly reduced hydroxyproline levels. Histopathological evaluation revealed that kidney injury increased collagen deposition in the UUO kidney (17.1 ± 0.43% collagen positive area, n=10) compared to the control kidney (2.0 ± 0.23%, n=10). TK850 treatment in UUO-P mice significantly attenuated collagen deposition (10.5 ± 0.38%, n=10), while UUO-I had significantly reduced collagen deposition as well (13.1 ± 0.25%, n=10). DISCUSSION/SIGNIFICANCE: Taken together, these results validate the dual TGFβR1/MAP4K4 inhibitor, TK850 as a potential therapeutic to mitigate renal fibrosis and supports the emergence of a combinational pharmacotherapeutic approach for multi-factorial kidney diseases.

OBJECTIVES/GOALS: In this work investigating the epidemiological differences associated with Post Acute Sequelae of SARS-CoV-2(PASC) patho genesis we willassess the sex differences inocular viral persistence and the immunologic profile in tear film obtained from COVID-19 patients. METHODS/STUDY POPULATION: Participants will be enrolled from the NIH funded RECOVER Consortium at the 15 adult hubs in the US and will include those with acute COVID-19 (n=250), followed up at baselineto 48 weeks post infection. RT-PCR will be used to detect viral RNA and electro chemiluminescence assays will be used to detect IgG, IgA1 and IgA2 antibodies.Tear film antibody titers will be measured longitudinally in all participants to assess the kinetics of the immune responses in those who developed PASC and those who did not. Tear film antibody titers will be correlated with antibody titers in the blood and compared between those individuals with or without measurable viral RNA in tear film. RESULTS/ANTICIPATED RESULTS: Logistic regression models will be used assess the association of viral persistence with PASC status controlling for relevant covariates. Linear mixed regression models will be used to assess the association of IgA1/IgA2 with PASC status. We expect to observe delayed clearance of viral RNA and elevation in SARS-CoV-2 specific IgA2/IgA1 in the tear film of patients with PASC compared with those without PASC. Given evidence of increased PASC risk in women we expect to observe higher rates of SARS-CoV-2 ocular viral persistence and higher SARS-CoV-2 specific IgA2/IgA1 ratios in women with COVID-19 when compared to men with COVID-19. DISCUSSION/SIGNIFICANCE: There is concern that PASC will pose a major global health challenge given the scale of the Covid-19 pandemicand the patho genesis remains unclear. This work is highly likely to improve our understanding of the mechanisms of PASC and the reasons why women are more vulnerable to this condition.

OBJECTIVES/GOALS: There is broad recognition that interdisciplinary research teams are uniquely suited to address complex research questions. At the Michigan Institute for Clinical & Health Research, we recognized a significant gap in support services at the University of Michigan for coordinating interdisciplinary teams to advance translational research. METHODS/STUDY POPULATION: The initial team science challenge we tackled was how to bring together cross-disciplinary groups, for the first time, to engage meaningfully and collaboratively with a ‘wicked’ problem of interest and create a shared vision. To address this, we developed Ideation Jams, which are facilitated experiences that help new groups build community, identify and prioritize research opportunities, determine how individual interests and other potential partners align with opportunities, and commit to next steps that will advance collaborative efforts. Ideation Jams leverage the methods and mindsets of design thinking, including divergence and convergence; making information visual; amplifying diversity; horizontal distribution of responsibility; and bias towards action. RESULTS/ANTICIPATED RESULTS: We have facilitated 11 Ideation Jams with 255 participants, including faculty, staff, health practitioners, and community members, who brought diverse expertise and insight to the research problems. Participant feedback has been overwhelmingly positive, with Ideation Jams fostering shared vision and innovation, and positively impacting various measures related to team performance. Participants have reported that Ideation Jams catalyzed various outcomes, including submission and award of grants, the introduction of new and specialized clinical offerings, and development of an interdisciplinary research agenda for their field of interest. Most recently, we trained representatives from five Clinical & Translational Science Award hubs to implement Ideation Jams at their universities. DISCUSSION/SIGNIFICANCE: Ideation Jams are ideal for mobilizing new groups around complex research problems, moving them from blue-sky thinking to action planning in three hours. Ideation Jams will be integrated into a suite of facilitated experiences, trainings, and grant development services to provide iterative support as teams advance their research priorities.

OBJECTIVES/GOALS: The goals of the present study were to examine levels and potential changes in oxytocin and HPL over the course of pregnancy. We assessed the potential predictive value of oxytocin and HPL on maternal-fetal attachment, anxiety and depression at three timepoints during pregnancy. METHODS/STUDY POPULATION: Pregnant women (n=70) enrolled in a longitudinal, rolling protocol study. Eligibility criteria included 1) singleton pregnancy confirmed at early pregnancy screen (EPS) ultrasonography, 2) mother aged 19 or greater, and 3) fluent in English. Predictors (oxytocin and HPL levels) were measured via blood draws at the same three times (early-stage, mid-stage, and late-stage) that MFA, anxiety and depression questionnaires were completed. RESULTS/ANTICIPATED RESULTS: An increased OT level compared to a mother’s average OT level did not have a statistically significant effect on MFA (within-person estimate = 0.02, 95% CI: -0.03 to 0.05, p = 0.427. An increased HPL level compared to a patient’s average HPL level did not have a statistically significant effect on MFA (within-person estimate = -0.10, 95% CI: -0.67 to 0.47, p = 0.730). The main effect of between-person HPL was significant; such that a one-unit increase in average HPL level was associated with a 0.52 higher anxiety score (between-person 95% CI: 0.08 to 0.96, p = 0.022). The main effect of between-person HPL was significant, such that an increased average HPL level was associated with a 0.45 higher depression score (between-person estimate = 0.45, 95% CI: 0.04 to 0.86, p=0.031). DISCUSSION/SIGNIFICANCE: To our knowledge, our study is the first to measure HPL and MFA over the course of a pregnancy. At this point, perhaps the best we can say is that HPL is a promising new target hormone that may be related to psychological symptoms surrounding pregnancy.

OBJECTIVES/GOALS: Healthcare sectors are rushing to develop AI models. Yet, a dearth of coordinated practices leaves many teams struggling to implement models into practice. The Enterprise AI Translation Advisory Board uses across-disciplinary team to facilitate AI translation. METHODS/STUDY POPULATION: The Mayo Clinic Enterprise AI Translation Advisory Board was established to assess AI solutions lever aging cross-disciplinary team science to accelerate AI innovation and translation. The 23-member board reflects expertise in data science, qualitative research, user experience, IT, human factors, informatics, regulatory compliance,ethics, and clinical care, with members spanning thought leadership, decision-making, and clinical practice. Taking an approach of respectful communication, transparency, scientific debate, and open discussion, the Board has consulted onover two dozen projects at various stages of the AI life cycle. RESULTS/ANTICIPATED RESULTS: Common issues identified for projects earlier in the AI life cycle, sometimes fatal but often address able once identified, include a lack of buy-in from potential product users, a lack of planningabout integration into clinical workflow, inadequately labeled data, and attempting to use machine learning when what is desired is really a causal model for intervening. Recommendations for projects later in the AI life cycle include details of a testing plan (silent evaluation, pragmatic clinical trials), advice about clinical integration, both post-hoc and on going auditing for performance disparities, and planning for regulatory clearance. DISCUSSION/SIGNIFICANCE: Advising is more valuable for projects at the ideation phase, when multi disciplinary interrogation can identify weaknesses. But at all phases, projects have gaps related to a lack of specific disciplinary expertise. A multi disciplinary cluster like the AI Translation Advisory Board seeks to address these gaps.

OBJECTIVES/GOALS: Healthcare organizations and payers are moving from accountability to effectiveness frameworks. Static vendor contracts for full-scale implementation limit organizations' ability to evaluate impact before scale-up, or to iteratively improve. Our team science innovation employs science and learning methods as systems engage vendors. METHODS/STUDY POPULATION: Our team science innovation is a method to assess and model impact of interventions at scale in healthcare delivery systems. We are integrating expertise in learning processes of an academic medical center (UCLA CTSI) with the organizational knowledge and methodological expertise of the nation’s largest Medicaid managed care plan (LA Care Health Plan), which has over 2 million members. The LA Care Advanced Analytics Lab has unique capability in machine learning, while enables deep learning of variation. Our innovative product is a template to quickly mobilize evaluation and learning for a diverse population in a varied and distributed delivery system. The template design enables rapid learning for the full-scale policy implementation often imposed by government, and in the short timeframes involved. RESULTS/ANTICIPATED RESULTS: LA Care and the UCLA CTSI partnered to provide subject matter expertise and design effective pilots for interventions such as transitional care services, complex care management, and physician home visit strategies, accounting for confounding factors affecting the intervention and outcome. So far, collaborative modeling and design has produced a successful pilot of a physician home visit program intended to reduce avoidable emergency department visits. This pilot quickly revealed several major changes that would need to be incorporated for the contracted vendor to produce results if operated at scale, further informed by machine learning, in sufficient time to inform the contracting process. There are multiple evolving applications, including housing/homelessness. DISCUSSION/SIGNIFICANCE: Integrating the large data and analytics of a large healthcare organization with learning methods from the CTSI -- including learning from variation and designs for studying impact during scale-up -- fosters academic-community team science that could significantly improve the value of our largest delivery systems, public and commercial.

OBJECTIVES/GOALS: Platelets interact with leukocytes in the circulation to modulate inflammation in chronic diseases. In previous clinical study, we showed that platelet leukocyte interaction is reduced in the circulation of patients with CKD. Preclinical studies are needed to show whether these findings are a precursor to or a result of CKD. METHODS/STUDY POPULATION: We used mouse models (wild type and platelet-defect) and induced CKD with intraperitoneal cisplatin injections. We measured platelet leukocyte interactions before and after CKD induction in the two models. RESULTS/ANTICIPATED RESULTS: We found platelet-leukocyte interaction to reduce after CKD induction in both wild type and platelet-defect mice. This coincided with a pro-inflammatory state in these mice, as measured by serum TNFalpha levels. Specifically, pro-inflammatory state was exacerbated in CKD of mice with platelet-defects compared to the wild type. DISCUSSION/SIGNIFICANCE: These findings recapitulate translational findings in human CKD samples and confirm that CKD state results in reduced platelet-leukocyte interactions in the circulation, and this change imparts a pro-inflammatory state in the CKD state.

OBJECTIVES/GOALS: To introduce the new Team Science Community Toolkit, co-created by community and academic partners, and showcase its potential to empower Community Organizations (COs) in achieving equity in community-engaged research (CER). METHODS/STUDY POPULATION: In response to the challenges faced by COs in CER collaborations, qualitative interviews were conducted with CO staff from historically marginalized communities. These interviews informed the development of the Team Science Community Toolkit, a collaborative effort involving a Community Advisory Board (CAB) and Team Science experts from Northwestern University. The toolkit, designed using a community-based participatory research approach, incorporates the Science of Team Science and User-Centered Design principles. Integrated into the NIH-sponsored COALESCE website, it includes templates, checklists, and interactive tools, along with a real-world simulation, to support COs in all stages of the research process. RESULTS/ANTICIPATED RESULTS: Focus groups and usability testing involving external community experts validated the toolkit’s content and usability. Participants expressed enthusiasm and a sense of empowerment, indicating that the toolkit allows them to actively shape research processes and infuse their specific voices and needs into their partnerships. The toolkit is designed to support breaking down barriers like jargon and cultural adaptability to improve accessibility and open conversation. The impact of this Team Science focused toolkit is under evaluation. This presentation will showcase the toolkit, detail its collaborative development, and explore potential applications, ultimately offering a path to more equitable and valuable community-based research. DISCUSSION/SIGNIFICANCE: By providing COs with the resources and knowledge to participate as equal partners in research collaborations, it enhances self-advocacy, transparency, and equity. The toolkit has the potential to utilize Team Science to foster productive communication in community-academic research partnerships.

OBJECTIVES/GOALS: We are using ethnographic methods and Dissemination and Implementation (D&I) frameworks to study barriers and facilitators to implementing ‘TeamMAPPS: Team Methods to Advance Processes and Performance in Science.’ TeamMAPPS is an evidence-based Team Science curriculum deployed as five online modules and being implemented across CTSA hubs. METHODS/STUDY POPULATION: For this pre-implementation study, we used the Implementation Mapping framework to understand likely barriers and facilitators, with the aim of designing implementation strategies and long-term outcome measures. Data included field notes from a two-day train-the-trainer, one visit to a key implementing site, and 27 interviews. Participants were four TeamMAPPS conceptualizers, four module designers, and 15 implementers from seven implementing sites, each with a CTSA hub (four were interviewed twice). We coded transcripts using the Consolidated Framework for Implementation Research (CFIR) to identify contextual barriers and facilitators to D&I, the Reach, Effectiveness, Adoption, Implementation, and Maintenance (RE-AIM) D&I outcomes framework, and target competencies of TeamMAPPS. RESULTS/ANTICIPATED RESULTS: Priority D&I outcomes that emerged were adoption, reach, and effectiveness. Potential barriers/facilitators to “adoption” included institutional willingness to incentivize scientists to utilize TeamMAPPS, support for Team Science at CTSAs, and systems of rewards for scientists to undergo trainings. Anticipated barriers/facilitators for “reach” were closely tied to adoption, such as institutions’ ability to persuade or require scientists to take trainings. Other issues relevant to reach included the time it takes to time to complete TeamMAPPS and potentially fraught intra-team dynamics arising if modules are implemented as a whole-team intervention. Anticipated barriers/facilitators for “effectiveness” included having adequate tools to assess actual impact. DISCUSSION/SIGNIFICANCE: TeamMAPPS has the potential to accelerate advances in translational sciences across the CTSA consortium. As this D&I study proceeds we will continue Implementation Mapping and use the Expert Recommendations for Implementing Change (ERIC) to develop bundles of implementer-informed strategies to the effectively deliver TeamMAPPS among CTSAs.

OBJECTIVES/GOALS: to investigate the potential impact of grandparental factors and multigenerational epigenetic inheritance on the development of ASD METHODS/STUDY POPULATION: Our study recruited participants from the CHARGE (Child Autism Risks from Genetics and the Environment) study, including grandparents, parents, and children. A questionnaire was used to gather information about the participants’ exposure to environmental factors. Saliva samples werecollected from 349 participants. Newborn dried blood spotsfrom probands and parents are still being collected from the California New born Registry. DNA was extracted from 349 saliva samples from 85 families and subjected to whole genome bisulfite sequencing (WGBS) to analyze DNA methylation. Sequence alignments and bioinformatic analyses will be performed using R packages called DMRichR and Comethyl. RESULTS/ANTICIPATED RESULTS: Sequence alignments and bioinformatic analyses are ongoing, utilizing DMRichR to identify individual genomic loci associated with ASD in each of the three generations and Comethyl to compare correlation patterns between methylation marks and selected variables, including grand parental exposures. New born blood spot collections of parents and probands are ongoing and will be used to identify potential ASD epigenomic signatures that are tissue and life-stage independent. DISCUSSION/SIGNIFICANCE: This research will provide new insights into the increased prevalence and underlying etiology of ASD that should pave the way for future research in the field. DNA Methylation signatures can help create molecular biomarkers which can be used together with behavioral clinical tests for diagnosis of ASD.