ABSTRACT IMPACT: Triple negative breast cancer (TNBC) affects 10-20% of women with breast cancer and is biologically more aggressive than other subtypes. The novel compound we have developed, DL7076, would give clinicians a vital strategy to improve the commonly used cyclophosphamide (CPA) and doxorubicin (DOX) regimen in the treatment of TNBC. OBJECTIVES/GOALS: The objective of this research project is to develop a novel compound which can activate both 1) the constitutive androstane receptor (CAR) and subsequently enhance the CYP2B6-mediated activation of CPA, and 2) the nuclear factor erythroid- related factor-2 (Nrf2) leading to the cardiomyocyte protection from DOX-associated cardiotoxicity. METHODS/STUDY POPULATION: Following the identification of the compound candidate, DL7076 was evaluated for tissue specific induction of CAR and Nrf2 using qPCR, western blot analysis, and luciferase reporter assays.

Further, we have developed a multicellular coculture model incorporating human primary hepatocytes for metabolism, TNBC spheroids as the target, and cardiomyocytes as a side target of DOX. We have investigated the anticancer effects of CPA/DOX on TNBC cells and the toxic effects on cardiomyocytes with/without a CAR-Nrf2 activator, in a multicellular environment where hepatic metabolism is well-retained. RESULTS/ANTICIPATED RESULTS: We found that our dual activator of CAR and Nrf2, DL7076, exhibits tissue specific induction of CAR and Nrf2. Inclusion of DL7076 in combination with the CPA/DOX regimen improves anticancer efficacy, through the subsequent increase in the formation of the active CPA metabolite. With the addition of DL7076, DOX-mediated off-target cardiotoxicity was markedly reduced.

Lastly, utilizing the novel coculture system with human primary hepatocytes, TNBC spheroids, and cardiomyocytes, the inclusion of DL7076 to the CPA/DOX regimen shows decreased spheroid viability and improved cardiomyocyte viability and function. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our findings suggest that DL7076 can facilitate DOX/CPA containing regimens by increasing CAR-mediated metabolism and subsequent CPA bioactivation while selectively protecting cardiomyocytes from DOX-induced toxicity. This research is expected to translate our basic scientific findings into therapeutic interventions for women with TNBC.

OBJECTIVES/SPECIFIC AIMS: Adults, 60 years of age and older, are in high demand for enrollment in many types of health research. Here we aimed to examine baseline, 60-day and 120-day follow-up trust in research and researchers of Floridians 60 years of age and older engaged in University of Florida’s HealthStreet community engagement initiative. METHODS/STUDY POPULATION: HealthStreet Community Health Workers (CHWs) assess health needs and trust in research of community members and screen for dementia, before providing medical and social services referrals and linkages to opportunities to participate in relevant health research at UF. In addition, participants are followed up at 60 and 120 days. RESULTS/ANTICIPATED RESULTS: Among the 2,193 older adults assessed by CHWs, 62.6% were female, 46.8% were African American, and 6.1% Hispanic/Latino. At baseline, 28.3% reported ever being in a research study; 7.7% reported not being interested in participating in research. Trust in research and researchers was high at baseline [scored from 1 to 10 where 10 was high; mean of 7.4 each for trust in research (SD=2.0) and trust in researchers (SD=2.1)] and high at both follow-ups [60 days 7.8 (SD=2.1) and 7.7 (SD=2) for trust in research and researchers respectively; 120 days 8.0 for both (SD=1.9 and 1.8 respectively)]. DISCUSSION/SIGNIFICANCE OF IMPACT: Individuals who are 60 and older have high trust in research and researchers when approached and high interest in research. Their trust continues through work with HealthStreet CHWs. Community engagement is an important part of the pipeline for recruitment of older adults for research.

OBJECTIVES/GOALS: The correction of spinopelvic parameters is associated with better outcomes in patients with adult spinal deformity (ASD). This study presents a novel artificial intelligence (AI) tool that automatically predicts spinopelvic parameters from spine x-rays with high accuracy and without need for any manual entry. METHODS/STUDY POPULATION: The AI model was trained/validated on 761 sagittal whole-spine x-rays to predict the following parameters: Sagittal Vertical Axis (SVA), Pelvic Tilt (PT), Pelvic Incidence (PI), Sacral Slope (SS), Lumbar Lordosis (LL), T1-Pelvic Angle (T1PA), and L1-Pelvic Angle (L1PA). A separate test set of 40 x-rays was labeled by 4 reviewers including fellowship-trained spine surgeons and a neuroradiologist. Median errors relative to the most senior reviewer were calculated to determine model accuracy on test and cropped-test (i.e. lumbosacral) images. Intraclass correlation coefficients (ICC) were used to assess inter-rater reliability RESULTS/ANTICIPATED RESULTS: The AI model exhibited the following median (IQR) parameter errors: SVA[2.1mm (8.5mm), p=0.97], PT [1.5° (1.4°), p=0.52], PI[2.3° (2.4°), p=0.27], SS[1.7° (2.2°), p=0.64], LL [2.6° (4.0°), p=0.89], T1PA [1.3° (1.1°), p=0.41], and L1PA [1.3° (1.2°), p=0.51]. The parameter errors on cropped lumbosacral images were: LL[2.9° (2.6°), p=0.80] and SS[1.9° (2.2°), p=0.78]. The AI model exhibited excellent reliability at all parameters in both whole-spine (ICC: 0.92-1.0) and lumbosacral x-rays: (ICC: 0.92-0.93). DISCUSSION/SIGNIFICANCE: Our AI model accurately predicts spinopelvic parameters with excellent reliability comparable to fellowship-trained spine surgeons and neuroradiologists. Utilization of predictive AI tools in spine-imaging can substantially aid in patient selection and surgical planning.

OBJECTIVES/GOALS: Pediatric heart failure is a complex disease presenting as an end-stage condition due to various etiologies and symptoms, causing 14,000 hospitalizations per year in the United States. Currently, there is a lack of objective metrics that are monitored non-invasively. This study explores tools to adapt remote monitoring technologies for pediatrics. METHODS/STUDY POPULATION: The task was determining commercially available and upcoming technologies for remote heart failure monitoring in the pediatric population. Literature and patent reviews were done in various databases with defined eligibility and key terms. Our inclusion criteria were: English peer-reviewed research or review articles, patents filed by cardiac monitoring companies, and independent groups focusing on non-invasive monitoring solutions. Key terms used for the literature search and patent review included pediatric heart failure’, at-home monitoring’, cardiac monitoring’, and non-invasive’. Based on a strong correlation between increased filling pressures and pediatric heart failure, the term filling pressures’ was also included in the search. RESULTS/ANTICIPATED RESULTS: Preliminary searches demonstrate an abundance of adult-use commercially available devices and patents for monitoring heart failure. Yet, there are no FDA-approved devices for use in the pediatric population. Current devices include monitoring metrics such as lung congestion and multi-parametric monitoring to capture heart rate, thoracic impedance, and oxygen saturation levels. In monitoring filling pressures, Abbott CardioMEMs is a leader in measuring pulmonary arterial pressure invasively in adults. Thus, there is a gap for non-invasive monitoring of filling pressures in both pediatric and adult populations. For further review, a larger focus will be placed on non-invasive methods for direct monitoring or extrapolation of cardiac filling pressures. DISCUSSION/SIGNIFICANCE: Current methods of heart failure monitoring are ineffective in serving the pediatric population. Thus, an identified gap exists in non-invasive filling pressure monitoring for pediatric heart failure. This review informs that innovation within this area would address inefficacies within the current paradigm of heart failure monitoring.

OBJECTIVES/GOALS: Tropsensor, a noninvasive portable device using infrared spectroscopy, delivers a troponin res ult within five minutes, significantly quicker than standard of care (SOC) assays. This pilot study assesses the correlation of the Tropsensor and high sensitivity cardiac troponin (hs-cTnI) assay results. METHODS/STUDY POPULATION: Patients undergoing cTnI testing with the Abbott Architect STAT (Abbott Laboratories) hs-cTnI assay were recruited at a quaternary-care emergency department (ED). The Tropsensor was applied to the underside of the patients’ wrist within 5 minutes of the SOC blood draw for 5 minutes. The results of the hs-cTnI assays were compared with the raw output of the Tropsensor device to assess the relationship using both Spearman’s and nonlinear logarithmic measures of correlation. Patient demographic data was extracted from the EHR to supplement the data collected for this study. RESULTS/ANTICIPATED RESULTS: 58 patients were recruited with a mean age of 60 years (60% male, 40% female). Due to connection error, 8 patients’data did not get captured by the device. Additionally, due to noise related to suboptimal device contact with the wrist, 24 patients’data (41%) were rejected. Of the 26 patients with usable data, 9 patients had a troponin above the 99th percentile threshold. A nonlinear correlation of 0.64 and Spearman’s rho of 0.59 were observed between the SOC hs-cTnI assay and Tropsensor optical data. DISCUSSION/SIGNIFICANCE: The Tropsensor exhibits 64.4% correlation to the SOC hs-cTnI assay. While further evaluation is needed, this exploratory study provides insight into the potential of a transdermal optical device to be used as a screening tool for AMI in an ED triage situation.

ABSTRACT IMPACT: Reversing tumor microenvironment (TME) immunosuppression will help to increase the overall efficacy of treatment of chemo-resistant triple negative breast cancer (TNBC) and mitigate racial disparities in treatment response. OBJECTIVES/GOALS: We have developed an ex-vivo whole tissue culture model to test the feasibility of reversing local immunosuppression in TME by chemokine modulatory (CKM) regimen. Our current objective is to analyze the molecular changes in CKM-treated chemoresistant TNBC from White and Black women and identify factors determining response to CKM. METHODS/STUDY POPULATION: Freshly resected residual TNBC from 20 White and 20 Black women ≥18 yrs old treated with neoadjuvant chemotherapy (NAC) will be procured. Tumor explants will be prepared & cultured in the absence and presence of CKM (Interferon-ð 〉1/4, TLR3 agonist rintatolimod and COX-2 inhibitor celecoxib). Chemokines implicated in cytotoxic T-lymphocyte (CTL)- & MDSC/ Treg attraction will be analyzed using Taqman & ELISA. We will have 80% power to detect a 0.7 standard deviation difference in chemokines between untreated & treated samples within and between cohorts using ANCOVA. Bulk RNA sequencing will be performed on both untreated & treated samples from CKM responding (highest aggregate increase in CTL- and highest decrease in Treg/MDSC-favoring chemokines in the top quartile) and non-responding (bottom quartile) tissues. RESULTS/ANTICIPATED RESULTS: Our preliminary data show that Black patients (pts) with breast cancer (BC) have an immunosuppressive TME associated with poor outcomes. This is similar to other existing literature showing that Black pts with BC have less favorable and more unfavorable chemokines in the TME. We anticipate the chemokine changes with CKM treatment will be larger in the Black cohort given their ability to elicit a robust inflammatory response. Therefore, we expect that CKM treatment will result in favorable TME in both groups and improve outcomes in TNBC, which has the worst prognosis of all subtypes, eliminating a key area of disparity in BC. The proposed transcriptome analysis will help identify key gene networks involved in response to CKM treatment and guide modulating the targets for non-responsiveness to improve efficacy of CKM. DISCUSSION/SIGNIFICANCE OF FINDINGS: Pts with residual disease (RD) after NAC have a 3-yr overall survival 68% vs. 94% for pts with complete response. Blacks have a higher incidence of TNBC with more likelihood of RD & mortality. We anticipate that the existing TME differences can be abrogated by our current CKM regimen or via developing an alternative CKM regimen optimized for Black pts.

OBJECTIVES/GOALS: A TIA is a transient episode of symptoms attributed to a cerebrovascular cause, and associated with an increased risk of stroke. Care of these patients often requires substantial resources in the Emergency Department (ED). We therefore described neuroimaging and hospitalization use for TIA within a nationally representative sample of US ED visits. METHODS/STUDY POPULATION: Retrospective cross-sectional analysis using TIA encounters in the 2018 National Emergency Department Sample (NEDS), an AHRQ dataset consisting of a weighted sample of 20% of all US ED encounters. Non-contrast Head CT, CTA Head, Carotid Ultrasound, MRI and CT Perfusion imaging utilization was determined based on Common Procedural Terminology (CPT) codes in the non-admitted encounters. The study population includes all adult patients with a discharge diagnosis of TIA as determined by ICD-10 codes (H34.0, G45.0-3, G45.8 G45.9) in any diagnosis position. The percentage of patients receiving each neuroimaging test was reported with the corresponding 95% confidence interval (CI). We utilized survey sample weights to generate reliable national estimates. RESULTS/ANTICIPATED RESULTS: The study population consisted of 80,803 ED encounters with a discharge diagnosis of TIA, representing 326,802 weighted ED visits nationally. Among this group, 46.8% of patients were discharged and 41.8% were admitted to the same hospital, 7% of patients were transferred to another facility, and the remaining 5% left AMA, were dispositioned to home health, died in the ED, or had an unknown disposition. Because discharged encounters retain their more precise CPT coding of procedural information, imaging analysis was conducted in discharged TIA encounters only. Of these encounters, 73% (95% CI, 70.7-76.5) received a noncontrast head CT, 20.9% (95% CI, 19.1-22.7) a CTA Head, 22.5% (95% CI, 20.6-24.4) a carotid ultrasound, and 31.5% (95% CI, 29.3-33.7) an MRI brain without contrast. DISCUSSION/SIGNIFICANCE: The discharge rate of just under 50% of patients is consistent with other published data, and represents a gradual trend over the past decade of decreased admissions for TIA. The fact that for many of these patients, the entire episode of care occurs in the ED setting suggests that the ED may be a rich target for future innovations in care for TIA.

OBJECTIVES/GOALS: Klotho is a protein linked to improved cognition in aging adults. A specific KLOTHO gene variant, KL-VS, increases circulating levels of Klotho. The current study aims to identify if the KL-VS haplotype and Klotho levels are associated with improved neurocognition in pediatric brain tumor survivors. METHODS/STUDY POPULATION: We are actively accruing pediatric brain tumor patients at UCSF alongside an existing multi-institutional cohort study investigating radiation-induced vasculopathies and cognitive outcomes in this population. Normal controls are being enrolled in parallel. Each patient undergoes: 1) single nucleotide polymorphism genotyping to identify KL-VS haplotype status, 2) enzyme-linked immunosorbent assays to measure circulating Klotho, 3) neurocognitive assessments with a computer-based, validated Cogstate battery, and 4) brain volume and white matter lesion segmentation analyses using MRI sequences obtained as part of routine care. RESULTS/ANTICIPATED RESULTS: Genotyping has been performed on 99 enrolled patients. KL-VS heterozygosity was seen in 22.7% of patients. To date, KL-VS status is not associated with neurocognitive outcomes at baseline or Year 1 testing. Association between KL-VS status, circulating Klotho levels, neurocognitive outcomes, brain volume and white matter lesion segmentation analyses is ongoing. We hypothesize that elevated Klotho levels will be associated with improved neurocognition, increased brain volumes in regions of interest and decreased white matter lesion volumes. DISCUSSION/SIGNIFICANCE OF IMPACT: If circulating Klotho leads to improved neurocognition in pediatric brain tumor survivors, Klotho levels might serve as a prognostic biomarker. Furthermore, as Klotho is being investigated for therapeutic indications, it may represent an intervention to prevent cognitive deficits in these patients.

OBJECTIVES/GOALS: Recent scientific discoveries show that human sex determination and differentiation is a spectrum of developmental processes and that sex/gender traits and identities not always fit binarity. This study aims to determine whether the visual arts can effectively transmit these complex scientific ideas on sex and gender variance. METHODS/STUDY POPULATION: A one-hour lecture for undergraduate students enrolled in a behavioral neuroendocrinology course included 17th century Spanish paintings and representative work from contemporary LGBTQIA+ artists in photography. Pre and post self-evaluation was conducted through five multiple choice questions. Chi Square test was employed for statistical analysis, which required elimination of no responses from item analysis. Statistical significance was defined as p value < 0.05. Three artistic images and two scientific images were included in the questions. Acquisition of content-specific knowledge on diverse body configurations, differences of sex development (DSD), transsexuality, histopathology of the gonad and neuroanatomy of the hypothalamus was evaluated. RESULTS/ANTICIPATED RESULTS: Twenty students completed the online pre- and post-test. We found that a 17th century Spanish painting was effective at making the distinction between transsexuality and intersexuality (p < 0.0005). Similarly, an artistic image that reinterprets the Vitruvius Man was effective for presenting the argument that diverse body configurations does not imply clinical pathology (p < 0.0001). Last, a scientific image showing the histopathology of an ovotestis was effective for introducing undergraduate students to diagnostic criteria for DSD (p < 0.02). No significant differences were found in the use of an artistic photograph depicting a young female to male transsexual individual nor in the use of a scientific image showing the neuroanatomical localization of the hypothalamus. DISCUSSION/SIGNIFICANCE: The learning of complex scientific concepts on human sex determination and differentiation can be affected by preconceived ideas, values, and attitudes towards sex and gender variance. The visual arts can provide a familiar ground of understanding between teachers and learners to transform such preconceptions.

OBJECTIVES/SPECIFIC AIMS: Wake Forest Baptist Health (WFBH) is an Academic Learning Healthcare System (aLHS) serving 24 counties in North Carolina and Virginia. Like many aLHSs, WFBH experiences strained community relationships attributable to a history of medical and research abuses against marginalized populations. This legacy accompanies longstanding community mistrust in the healthcare system and research. To overcome these challenges, community-engaged research (CEnR) approaches have potential to repair community-academic relationships, improve public health, and empower groups that traditionally have been neglected by or overlooked in research. To develop and revise our understanding of how CEnR is harnessed at WFBH, semi-structured interviews were conducted with investigators and study staff experienced in CEnR approaches. In-depth interview guides were designed iteratively to capture socio-contextual and detailed descriptions of perceptions, experiences, and strategies specific to the use of CEnR. METHODS/STUDY POPULATION: A keyword search performed within WFBH study records identified 51 investigators whom had submitted research proposals related to CEnR within the past ten years. Sixteen were confirmed eligible based on a review of proposal abstracts, of which 14 responded to email invitations agreeing to participate. Four additional participants were referred by initial participants. Eighteen investigators (16 faculty and 2 research associates) provided consent and completed Interviews. RESULTS/ANTICIPATED RESULTS: The participant sample was 50% female with a mean age of 55 years, 11% Black and 89% White, with representation across various academic backgrounds (e.g., anthropology, medicine, psychology, and public health) A majority of participants (89%) hold doctoral degrees (i.e., PhD, DrPH, EdD, MD, and MD-PhD). On average, participants had been employed at WFBH for 13.9 years, and represented various departments including dermatology, epidemiology and prevention, family medicine, neurology, social sciences and health policy, and psychiatry. Nearly all participants (89%) indicated they had never received formal education or training in CEnR, though 100% reported “on-the-job” training in CEnR. Interviews were audio-recorded, transcribed, coded, and analyzed following an inductive thematic approach, from which twenty-two themes emerged across six domains related to CEnR (Table 2), including: Conceptualization and Purpose, Value and Investment, Community-Academic Partnerships, Sustainability, Facilitators, and Challenges. Results also provided key characteristics that define CEnR (Table 3), and yielded 11 emerging needs necessary to enhance CEnR within aLHSs (Table 4). DISCUSSION/SIGNIFICANCE OF IMPACT: The results of this study provide information critical to understanding how CEnR frameworks and approaches can be harnessed not just in Schools of Public Health, but within aLHSs to build and repair community-academic partnerships, inform research and institutional priorities, and address community health concerns. Despite the small sample size, the number of participant interviews was sufficient to achieve saturation while also providing broad and unique perspectives across various fields and CEnR approaches. Overall, participants conceptualized the purpose and goals of CEnR quite similarly, though there was a great deal of variance in how CEnR was defined and operationalized across interviews, indicating a need to more clearly articulate important features that enhance understanding of what CEnR is and what it is not (Table 3). These discrepancies and inconsistencies indicate a potential need for additional formal training in the understanding and use of CEnR approaches, which is supported by the fact that nearly all participants reported receiving no formal training in CEnR. Across all interviews, participants expressed a need for health care providers and researchers to better understand community contexts, social determinants of health, and historical factors influencing community health and participation in research (Table 4). This work and the data presented here are important for informing CEnR approaches and will be useful for guiding the development of a model incorporating the core tenets of CEnR within the mission, vision, and priorities of aLHSs.

OBJECTIVES/GOALS: The rates of computational phenotyping algorithm reuse across health systems are low, leading to a proliferation of algorithms for the same trait. We propose a framework for reusing computational phenotyping algorithms and describe the real-world deployment of this framework for the development of the Colorado Diabetes EHR Research Repository. METHODS/STUDY POPULATION: The novel phenotype reuse framework consists of 4 steps: select algorithms that are appropriate for reuse by assessing whether they are fit for purpose; extend the algorithm to account for changes in data and care practice standards; localize the algorithm to use local database standards and terminologies; optimize the algorithm by applying a data driven approach to achieve the desired local performance. To identify individuals with type 1 diabetes (T1D) or type 2 diabetes (T2D), we selected and implemented T2D algorithms in a cohort of adults with any diabetes or pre-diabetes related diagnosis code, medication, or abnormal glucose-related laboratory test in the clinical data warehouse for UCHealth and the University of Colorado. RESULTS/ANTICIPATED RESULTS: We included a total of 926,290 patients who were identified by initial filters. Patients were more likely to be female (53%), identify as non-Hispanic white (69%) and had a median age of 58 years (IQR: 41, 70). Implementation, extension, localization, & optimization through iterative chart review prioritized high sensitivity for all-cause diabetes and high specificity for T1D and T2D. Of the original cohort, 252,946 (27%) were identified by the all-cause diabetes algorithm. Of these 11,688 were identified as T1D and 135,588 as T2D. After optimization the all-cause diabetes algorithm had 88% sensitivity, 90% specificity, 74% positive predictive value (PPV), and 96% negative predictive value (NPV). Our algorithms for T1D and T2D had high specificity (100% and 99%, respectively) and PPV (100 and 96% respectively). DISCUSSION/SIGNIFICANCE: Developing computational phenotyping algorithms is expensive and time consuming, yet algorithm reuse is low due to a lack of practical approaches for reusing algorithms. We demonstrate application of a novel framework for algorithm reuse, yielding good alignment of algorithm performance with study goals for identifying individuals with diabetes.

OBJECTIVES/GOALS: The aim of this study was to define the transcription profiles of the molecular markers of mitochondrial biogenesis and fusion/architecture, and the markers of mtDNA copy numbers in the peripheral blood mononuclear cells (PBMCs) from war veterans with/without post-traumatic stress disorder (PTSD). METHODS/STUDY POPULATION: The peripheral blood mononuclear cells (PBMCs) from war veterans with/without post-traumatic stress disorder (PTSD) were used to monitor transcription profile of the molecular markers of mitochondrial biogenesis and fusion/architecture, as well as the markers of mtDNA copy numbers. The human male immortalized monocytes were exposed in vitro to hormonal markers of PTSD in order to monitor the effects of each particular hormonal marker on the molecular markers of mitochondrial biogenesis and fusion/architecture, as well as the markers of mtDNA copy numbers. RQ-PCR analyses were used to define transcriptional profile of above mentioned markers. RESULTS/ANTICIPATED RESULTS: The transcription profiles of above mentioned markers were disturbed, with high individual variability within the groups. A significant increase in the expression of the PPARGC1A transcript was observed in a group of subjects with current PTSD, as well as in the subjects with “life-time” PTSD, compared to healthy controls. PPARGC1B, NRF2 and MFN2 transcripts increased only in PBMCs of “life-time"-PTSD, while the level of transcripts for other investigated genes and the ratio of markers of mtDNA copy numbers showed no significant difference between groups. The in vitro results showed parallelism with the results obtained using the PBMCs from the subjects of the PTSD study. DISCUSSION/SIGNIFICANCE OF IMPACT: Although preliminary (the analysis require a larger number of subjects), the results are first findings and a solid base for further extensive multidisciplinary research in order to clarify the molecular mechanisms for the prevention and treatment of trauma-induced pathological conditions.

The Translational Science TS22 conference in Chicago in April 2022 was the first time post-pandemic that members of the Association of Clinical and Translational Science were able to meet up in person to share scientific advances. Given the remaining level of risk due to COVID-19, the meeting was designed as hybrid allowing virtual participation to some of the presentations. Prior to the meeting, JCTS Junior Editors were invited to report on the plenary sessions of the meeting. The present perspective constitutes a summary of three plenary sessions.

The COVID-19 pandemic raised the importance of adaptive capacity and preparedness when engaging historically marginalized populations in research and practice. The Rapid Acceleration of Diagnostics in Underserved Populations’ COVID-19 Equity Evidence Academy Series (RADx-UP EA) is a virtual, national, interactive conference model designed to support and engage community-academic partnerships in a collaborative effort to improve practices that overcome disparities in SARS-CoV-2 testing and testing technologies. The RADx-UP EA promotes information sharing, critical reflection and discussion, and creation of translatable strategies for health equity. Staff and faculty from the RADx-UP Coordination and Data Collection Center developed three EA events with diverse geographic, racial, and ethnic representation of attendees from RADx-UP community-academic project teams: February 2021 (n = 319); November 2021 (n = 242); and September 2022 (n = 254). Each EA event included a data profile; 2-day, virtual event; event summary report; community dissemination product; and an evaluation strategy. Operational and translational delivery processes were iteratively adapted for each EA across one or more of five adaptive capacity domains: assets, knowledge and learning, social organization, flexibility, and innovation. The RADx-UP EA model can be generalized beyond RADx-UP and tailored by community and academic input to respond to local or national health emergencies.

OBJECTIVES/GOALS: With a growing intеrеst in tailoring disеasе diagnosis to еach individual as opposеd to a “onе-sizе-fits-all” approach, our aim is to еnhancе thе robustnеss of thе Itеrativе Clustеring for Subtypе Discovеry (iKCAT) algorithm in charactеrizing lung cancеr subtypеs for individualizеd trеatmеnt. METHODS/STUDY POPULATION: Our mеthod еxplorеs thе robustnеss of thе prеviously dеvеlopеd iKCAT algorithm. This itеrativе clustеring mеthod finds robust—homogеnеous and diffеrеntiablе—subtypеs of lung nodulеs through itеrativе K-mеans clustеring that hеlps classify thеm and lеavеs somе data unclustеrеd. This sеt of unclustеrеd or “hard” data rеprеsеnts imagеs that cannot confidеntly bе assignеd to any subtypеs and may rеquirе morе rеsourcеs (е.g., timе or radiologists) to diagnosе. Wе еxplorе thе robustnеss of iKCAT across multiplе fеaturе spacеs, including dеsignеd imagе fеaturеs (which arе еnginееrеd to capturе somе propеrtiеs such as lеvеl of еlongation, еccеntricity and circularity), rеducеd dеsignеd imagе fеaturеs using Principal Componеnt Analysis (PCA) and Uniform Manifold Approximation and Projеction (UMAP). RESULTS/ANTICIPATED RESULTS: Whеn running our еxpеrimеnt on thе 64 imagе fеaturеs, our rеsults consistеntly carvеd out a singlе purе, homogеnеous clustеr ovеr thе coursе of 30 iKCAT runs. From an initial datasеt of 1490 data points, 1430 points wеrе lеft unclustеrеd in this fеaturе spacе. Whеn conducting thе 30 iKCAT runs on thе PCA fеaturе spacе with 10 componеnts, wе found it did not producе any distinct clustеr abovе thе dеfinеd homogеnеity thrеshold. Thе 2D UMAP fеaturе spacе consistеntly gеnеratеd 8 clustеrs with an avеragе homogеnеity of 87. 22% ovеr 30 runs, and only lеft 9 points unclustеrеd. Ovеr 30 iKCAT runs, wе idеntifiеd 8 pеrsistеnt clustеrs or subtypеs, 3 mostly malignant and 5 mostly bеnign clustеrs. DISCUSSION/SIGNIFICANCE: Through our еxpеrimеnt using thе iKCAT algorithm, wе found that iKCAT’s clustеring functionality producеd thе most pеrsistеnt rеsults on thе 2D UMAP fеaturе spacе duе to its high avеragе homogеnеity scorеs and consistеncy in idеntifying clustеrs/subtypеs, hеlping improvе tailorеd disеasе diagnosis.

ABSTRACT IMPACT: Indiana CTSI Team Science to maximize rapid collection, analyses and dissemination of biosamples collected from patients with Covid-19 to provide preliminary data for grant applications on the pathogenesis and outcomes of patients with Covid-19. OBJECTIVES/GOALS: When Covid-19 hit Indiana in April, there was an immediate need to respond rapidly to coordinate research across our healthcare systems. The CTSI became a point of contact for coordinating research endeavors including activation of clinical trials and use of precious samples from patients with Covid-19 to maximize preliminary data for grants. METHODS/STUDY POPULATION: The Indiana CTSI coordinated collection of biospecimens at multiple hospitals using in person and remote consenting via telephone or on a smartphone utilizing a QR code. We also retrieved existing samples from the Indiana Biobank previously collected for future research and from subject positive for Covid-19 by search of the linked electronic health record (EHR). A total of 224 subject samples (7 children, 36 previously collected, and 6 with both acute and recovered specimens) were obtained over a four month period. Our CTSI cores ran varied analyses collated to a single database, linked to the EMR for use as preliminary data for grant applications to avoid redundancy of measures on limited samples. RESULTS/ANTICIPATED RESULTS: The 224 subject samples were used for whole exome DNA sequencing, RNA seq, analyses of 48 plasma cytokine/chemokines by multiplex analyses, and PBMC isolated for culture and assessment of secreted cytokines. The clinical data were linked and included demographics, hospitalization length of stay and need for mechanical ventilation, max and min oxygen levels, liver function tests, IL-6, D-dimer, CRP, LDH, and ferritin, need for dialysis, and echocardiography. Additional clinical data were available upon request. A survey was sent to our CTSI email to query for potential interest in the data with 87 inquiries, and to date 46 investigators have requested data and/or additional samples. DISCUSSION/SIGNIFICANCE OF FINDINGS: During the first surge of Covid-19, the CTSI coordinated analyses for the dissemination of results for use by CTSI investigators to minimize duplication of assays and increase availability. The collaboration of research coordinators, biobank, research cores, and informatics demonstrates the power and agility of team science in the Indiana CTSI.

OBJECTIVES/GOALS: The Title V Coop developed CRESCO, a physical and virtual space in the libraries of the two cooperating institutions. Adopting a flexible and transformational approach, it offers services to support the development of research and information skills of undergraduate students and faculty who receive clinical-translational research (CTR) training. METHODS/STUDY POPULATION: Since 2016, CRESCO has been staffed by a multidisciplinary team composed of three librarians, a statistician, an instructional designer, and an IT specialist. The physical facilities of the two libraries were remodeled and equipped, and a central portal was created to provide services and access to resources on a 7/24 basis. Online tutorials, workshops, and mentoring services have been offered that address topics in statistics, literature search, plagiarism, and the use of several research software. Services statistics are collected, and a questionnaire is administered to evaluate the workshops. RESULTS/ANTICIPATED RESULTS: The main results include 12 online tutorials created in CTR areas and available in the CRESCO hub portal; 14,660 mentoring/consultations offered in statistics, the use of research-related software, and the search for scientific literature search; and 6 online workshops created in CTR areas, with 463 attendees. When evaluating online workshops, participants considered that their acquired learning was high or extremely high on the following topics: use of Intellectus Statistics (88%, n = 96); selection of statistical tests (81%, n = 92); use of Turnitin (85%, n = 76); literature search (91%, n = 58); and citations and references in Mendeley (90%, n = 67). DISCUSSION/SIGNIFICANCE: These results suggest that the flexible, multidisciplinary, and transformational approach of CRESCO has been successful in helping undergraduate students and faculty develop the skills necessary to conduct CTR projects.

OBJECTIVES/GOALS: The purpose of this group is to foster professional and personal growth as leaders, provide peer mentoring, integrate the roles of scientist, woman, and mother, and build a self-sustaining network of peers for ongoing support throughout their careers in an academic setting. METHODS/STUDY POPULATION: - Study population: 12 Health sciences research faculty (50% protected research time); identify as female; mother of school-age and/or younger children Methods: Year-long program; including a 2-day retreat based on Brene Brown's Dare to Lead RESULTS/ANTICIPATED RESULTS: - Despite the pandemic, 100% of participants continued in the program over the one-year duration and met the attendance requirement of 75% - Screening for burnout was effective - facilitator was able to intervene when severe burnout was noted.

OBJECTIVES/GOALS: Multiple Organ Failure, often precipitated by Acute Lung Injury, is a life-threatening condition that causes death in military and civilian life. Furthermore, Acute Kidney Injury is very common, increasing morbidity and mortality rates. Therefore, understanding the molecular difference between survivors and non-survivors is urgently needed. METHODS/STUDY POPULATION: A 24-hour unilateral pulmonary contusion porcine model (pneumonectomy) of trauma-induced Multiple Organ Failure (MOF) model (n=17) and separate 48-hour polytrauma injury of bilateral pulmonary contusion, traumatic brain injury, and hemorrhage (polytrauma) MOF model (n=26) was developed at Dr. Batchinsky's AREVA laboratory. Serum was assayed at baseline and 3h or 6h post-trauma for amino acid metabolites using the Zip-Chip platform for mass spectrometry. The IDO1 enzyme activity assay kit (ab235936) was used to measure IDO1 enzyme activity in the tissue. Mass Spectrometry Imaging (MSI) was employed to frozen kidney tissues. Tissues were sectioned to 10- micron thickness. For MSI, the DAN matrix was utilized, and MALDI-MSI images were processed and obtained from METASPACE and SCILS software. RESULTS/ANTICIPATED RESULTS: In the pneumonectomy model, 10 survived, 7 died, and in the polytrauma group, 13 survived, and 13 died. In the pneumonectomy model, there was a significant increase in the serum kynurenine/tryptophan (KYN/TRP) ratio in the non-survivors 3h post-injury. A similar pattern was found in the validation group, which showed a significant increase in the KYN/TRP ratio at 6h post-trauma in non-survivors from the polytrauma model. There was a significant increase in IDO1 enzyme activity in non-survivor kidney tissues and a downregulation of tryptophan (TRP) metabolite in the kidney section in the non-survivor group post-trauma. DISCUSSION/SIGNIFICANCE: An increase in the KYN/TRP ratio post-trauma identified the pigs that suffered early mortality. A decrease in TRP metabolite and an increase in IDO1 enzyme activity in the kidney could contribute to an increase in KYN in the non-survivors. As a result, focusing on therapeutics targeting the KYN/TRP to reduce the incidence and severity of MOF is warranted.