The Implementation Science Centers in Cancer Control (ISC3) initiative, funded by the National Cancer Institute, called for the development of implementation laboratories to bolster implementation science, create research-ready environments, and expedite adoption and implementation of evidence-based interventions (EBIs) into practice. The Building Research in Implementation and Dissemination to close Gaps and achieve Equity in Cancer Control (BRIDGE-C2) Center is one of seven ISC3 centers. BRIDGE-C2 aims to identify strategies to improve implementation of cancer prevention EBIs and conduct research / develop pragmatic methods to tailor, enhance, and support the adoption and sustainability of these strategies; advance implementation science; and build capacity and training opportunities. Since its inception, the BRIDGE-C2 Center has been conducting research and training activities to advance knowledge on how to effectively implement strategies to improve cancer prevention EBIs in primary care clinics serving socioeconomically disadvantaged patients. The translational science benefits model (TSBM) provides a useful framework for organizing a description of the BRIDGE-C2 Center’s activities. In this paper, we describe examples of BRIDGE-C2 activities and the specific impact indicators within each relevant domain/subdomain of the TSBM, demonstrating that a single activity or project has multiple impacts on methods and capacity building, clinical domains, and community health.

The pervasive problem of irreproducibility of preclinical research represents a substantial threat to the translation of CTSA-generated health interventions. Key stakeholders in the research process have proposed solutions to this challenge to encourage research practices that improve reproducibility. However, these proposals have had minimal impact, because they either 1. take place too late in the research process, 2. focus exclusively on the products of research instead of the processes of research, and/or 3. fail to take into account the driving incentives in the research enterprise. Because so much clinical and translational science is team-based, CTSA hubs have a unique opportunity to leverage Science of Team Science research to implement and support innovative, evidence-based, team-focused, reproducibility-enhancing activities at a project’s start, and across its evolution. Here, we describe the impact of irreproducibility on clinical and translational science, review its origins, and then describe stakeholders’ efforts to impact reproducibility, and why those efforts may not have the desired effect. Based on team-science best practices and principles of scientific integrity, we then propose ways for Translational Teams to build reproducible behaviors. We end with suggestions for how CTSAs can leverage team-based best practices and identify observable behaviors that indicate a culture of reproducible research.

Research articles in the clinical and translational science literature commonly use quantitative data to inform evaluation of interventions, learn about the etiology of disease, or develop methods for diagnostic testing or risk prediction of future events. The peer review process must evaluate the methodology used therein, including use of quantitative statistical methods. In this manuscript, we provide guidance for peer reviewers tasked with assessing quantitative methodology, intended to complement guidelines and recommendations that exist for manuscript authors. We describe components of clinical and translational science research manuscripts that require assessment including study design and hypothesis evaluation, sampling and data acquisition, interventions (for studies that include an intervention), measurement of data, statistical analysis methods, presentation of the study results, and interpretation of the study results. For each component, we describe what reviewers should look for and assess; how reviewers should provide helpful comments for fixable errors or omissions; and how reviewers should communicate uncorrectable and irreparable errors. We then discuss the critical concepts of transparency and acceptance/revision guidelines when communicating with responsible journal editors.

ABSTRACT IMPACT: We devised a new method to produce highly potent SARS-CoV2-specific that can be used to treat severely ill patients with Covid-19. OBJECTIVES/GOALS: Neutralizing antibodies against SARS-CoV-2 are thought to offer the most immediate and effective treatment for those severely afflicted by Covid-19. We devised an approach for rapid and efficient generation of human monoclonal antibodies with neutralizing activity against SARS-CoV-2. METHODS/STUDY POPULATION: SARS-CoV-2 S1 spike protein-specific memory B cells were isolated from 12 subjects recovering from infection with that virus. Paired end single index sequencing was performed using up to 10,000 antigen-specific B cells per subject. Antigen-specific B cell clones were identified by unique diversity and joining gene V(D)J rearrangements and the CDR3 regions. VH and VL regions were cloned and the products expressed in 293T/17 cells to generate spike-specific human monoclonal antibodies. RESULTS/ANTICIPATED RESULTS: Forty-three human monoclonal antibodies were produced. Every monoclonal antibody so generated neutralized viruses pseudotyped with Spike protein of the Wuhan-1 strain. Eighteen monoclonal antibodies neutralized pseudotyped viruses with half-maximal inhibitory concentration (IC50s) between 1 pg/mL and 1 ng/mL (6.7 x 10E-15 M to 6.7 x 10E-12 M), exceeding by 10-100-fold the potency of previously reported anti-SARS-CoV-2-neutralizing monoclonal antibodies. Eight monoclonal antibodies neutralized viruses pseudotyped with mutant spike proteins previously identified in clinical isolates, including receptor binding domain mutants and the C-terminal D614G mutant with IC50<6.7 x10E-12M. DISCUSSION/SIGNIFICANCE OF FINDINGS: We show that SARS-CoV-2 evokes high affinity B cell responses. Some B cells produce antibodies that are broadly neutralizing; others produce strain-specific antibodies. However, antigenic variants that would potentially escape control by immunity or vaccination were nonetheless identified.

ABSTRACT IMPACT: Development of our animal model of moyamoya will provide a meaningful assessment of therapeutic efficacy of interventions applicable to the clinical setting. OBJECTIVES/GOALS: Moyamoya is a cerebrovascular condition with progressive stenosis of the internal carotid arteries (ICA) and formation of abnormal vascular collaterals at the base of the brain, all of which result in ischemic and hemorrhagic strokes. We aim to develop a needed animal model of this condition in order to develop new therapeutics. METHODS/STUDY POPULATION: Male and female C57Bl/6J mice (4 months old) underwent surgery for the unilateral placement of a microcoil (0.16 mm ID) onto the proximal ICA or sham control. After 30 and 60 days (N = 6-8/time point), the brain blood vessels were examined for changes in diameter, number of anastomoses, and development of new collaterals using DiI stain. Brain tissue was examined for micro-hemorrhages using Prussian blue stain, and cross-sections of blood vessels were examined for intimal thickening using H&E and smooth muscle actin. Expression of vascular endothelial growth factor (VEGF), which is associated with angiogenesis and moyamoya syndrome, was quantified by qPCR. Blood samples were also analyzed for inflammatory biomarkers using ELISA. RESULTS/ANTICIPATED RESULTS: Within 30 days, the distal ICA and anterior cerebral artery (ACA) had significantly decreased diameters at the Circle of Willis, with an initial decrease in the number of cortical anastomoses. Histology demonstrated smaller lumen diameter and alterations to in the various layers of the blood vessels, indicating intimal thickening and stenosis of the affected blood vessels. There was also a significant increase in the number of intracranial micro-bleeds, suggesting a compromised vascular integrity. This may be due, in part, to a significant upregulation in VEGF gene expression within the striatum, a region of hemorrhagic occurrence in moyamoya patients. DISCUSSION/SIGNIFICANCE OF FINDINGS: We report the development of an animal model with vasculopathies that mimic those observed in patients with moyamoya syndrome. With further characterization, this animal model will have a positive impact as a meaningful assessment of therapeutic efficacy of interventions applicable to the clinical setting.

OBJECTIVES/SPECIFIC AIMS: Parkinson’s disease (PD) is a condition that affects over a million Americans, and despite current medical therapies, the progression of the disease results in impaired generation of internally timed or guided (IG) movements. To address this loss of motor function, previous rehabilitation therapies have focused on remediating the affected striatal-thalamic-cortical circuits (STC), primarily thought to be responsible in generating timed motor patterns. However, given the disease leads to the cell death of dopaminergic cells that are essential for proper STC function, we propose a motor therapy aimed at utilizing a compensatory parallel cerebellar-thalamic-cortical (CTC) pathway, recruited to perform externally guided (EG) movements, in which gait initiation is driven from sensory input. Our previous study has shown efficacy in our novel argentine tango therapy and improves behavioral measures above the relevant MCID threshold, but it has not been established that the CTC are in the causal pathway that are responsible for these changes. Using neural measures from task fMRI, we have begun to characterize networks that have changed and quantify any associations with behavioral metrics. METHODS/STUDY POPULATION: Patients were randomly assigned to an IG (n=18), EG (n=18), or education contact control (n=14). Participants were assessed preintervention and postintervention for behavioral motor and cognitive measures and neurophysiologically with task based fMRI. In the task, participants performed a foot tapping task under both IG (tap their foot in previously learned rhythm) or EG (tap immediately after receiving a tactile cue on their hand) conditions. The fMRI data were preprocessed using AFNI and registered to MNI standard space. The brainnetome atlas was applied and the average time series of each region of interest (ROI) was used to increase the signal to noise ratio. The activation of these ROI with respect to the stimulus was modeled using GLM, and we estimated the area under the curve during the task blocks. A 1-way ANOVA analysis on these betas were performed between the pre and the post intervention time points and the ROIs that were above a significance of 0.95 were identified and corrected for multiple comparisons. The change in beta in all ROIs for each individual were calculated and then correlated with the changes in the behavioral data, to see which changes in ROI areas matched the best with the behavioral changes. RESULTS/ANTICIPATED RESULTS: The EG group showed significant changes only in the EG task in 2 areas—inferior frontal gyrus and inferior temporal sulcus. Correlating to the cognitive behavioral measures show reduced error from the Inferior frontal gyrus (corr>0.5) best reflect changes in observed. There were no changes to either the STC or the CTC pathways. The IG group showed no changes behaviorally and showed no changes neurally as well. The control group showed no changes behaviorally, but neuronally certain DMN nodes, such as the precuneus and inferior temporal regions showed a significant change for both tasks. DISCUSSION/SIGNIFICANCE OF IMPACT: Addressing the damaged STC pathway directly through IG therapy may not be effective. The EG therapy may not be able to enhance the STC pathway. However, the therapy appears to utilize new areas in the frontal regions and correlates with positively with changes in spatial memory and balance tasks. Contrary to our hypothesis the CTC circuit was not upregulated for performance of the IG or EG task, but therapy may have enhanced recruitment of other cognitively engaged areas. The educational control group interestingly showed changes in the DMN network, which has been shown to be linked to attention during tasks blocks.

OBJECTIVES/GOALS: We evaluated the long-term success of tissue engineered intervertebral discs (TE-IVDs) cultured in flexible (FPLA) or stiff (PLA) support materials, hypothesizing that FPLA would maintain disc height and tissue hydration in the minipig spine. METHODS/STUDY POPULATION: TE-IVD: NP cells were encapsulated in alginate and NP plugs were placed in the center of FPLA cages. AF cells were encapsulated in type I collagen and pipetted around NP plugs. Implantation: Empty FPLA cages (n=4), and TE-IVDs cultured in FPLA (n=4) were implanted at C3-4 or C5-6 following complete discectomy (DX) in skeletally mature minipigs (n=4). Imaging and Quantification: Terminal disc height indices (DHI) were calculated from weekly x-rays using a previously described method, and results were compared to the PLA pilot study. T2 MRI scans were taken of levels treated with TE-IVDs to quantify disc hydration as previously described. RESULTS/ANTICIPATED RESULTS: FPLA cages restored DHIs to native levels until endpoint. In contrast, PLA cages fractured, and terminal DHIs were statistically similar to DX levels. Of the four levels treated with TE-IVDs, 2 were displaced from the disc space. Stabilized levels yielded DHIs which were statistically similar to native IVD and greater than displaced and DX levels. Displaced levels yielded DHIs which were significantly lower than native and stabilized levels, but greater than DX levels (P<0.05). T2 MRIs of stabilized TEIVDs revealed that levels treated with a construct maintained tissue hydration which was significantly greater than levels treated with an empty cage or DX levels (P<0.0001), but which was about half the hydration of native disc tissue. DISCUSSION/SIGNIFICANCE: Implanting TE-IVDs with FPLA support cages leads to disc height maintenance and the stabilization of hydrated tissues in the spine, enhancing the long term success of TE-IVD implants and providing a basis for clinical translation.

OBJECTIVES/SPECIFIC AIMS: The objective of this project is the application of complex fusion models, which combine observed and modeled data, to areas with sparse monitoring networks with multiple chemical components is under-developed. Such models could provide improved accuracy and coverage for air quality measurement predictions, an area greatly limited by the amount of missing data. METHODS/STUDY POPULATION: This project focuses on the development of methods for improved estimation of pollutant concentrations when only sparse monitor networks are found. Sparse monitoring networks are defined as areas where fewer than three criteria air pollutants (based on EPA standards) are monitored. Particularly, a multivariate air pollutant statistical model to predict spatio-temporally resolved concentration fields for multiple pollutants simultaneously is developed and evaluated. The multivariate predictions allow monitored pollutants to inform the prediction of nonmonitored pollutants in sparse networks. RESULTS/ANTICIPATED RESULTS: Daily, ZIP code level pollutant concentration estimates will be provided for 8 pollutants across South Carolina, and goodness of fit metrics for model variants and previously established methods will be compared. DISCUSSION/SIGNIFICANCE OF IMPACT: These methods utilize only widely available data resources, meaning that the improved predictive accuracy of sparsely monitored pollutant concentrations can benefit future studies in any US area by improving estimation of health effects and saving resources needed for supplemental air pollutant monitoring campaigns. Our method for estimation attempts to improve predictive accuracy and data availability for sparsely monitored pollutants and areas.

OBJECTIVES/GOALS: Aim 1: To explore methodologies to identify gender minorities, and Filipinos, separately, in an electronic health record system. Aim 2: To characterize the similarities and differences in demographic, socioeconomic, and mental health of Filipinos gender minorities compared to Native Hawaiian Pacific Islanders, and White/ European Americans. METHODS/STUDY POPULATION: This study was approved by the University of Hawaii Institutional Review Board. Cross-sectional retrospective data were obtained from a collaborative community clinic’s electronic health record system. Patients were age 18 and older with a clinical diagnosis for gender dysphoria and from Native Hawaiian Pacific Islander, White/ European American, and Filipino backgrounds. RESULTS/ANTICIPATED RESULTS: Preliminary data revealed that 11% of the clinical population were diagnosed with gender dysphoria (N=373) with 57.6% (n=215) who met the inclusion criteria with complete health registration forms. Patients were from Filipino (21.8%), Native Hawaiian Pacific Islander (23.3%), White/ European American (31.6%), and multiethnic (23.3%) backgrounds. Most patients reported mental health (e.g., depression) conditions (50.6%-64.7%). Further statistical analyses will reveal if Filipinos have higher or lower levels of anxiety, depression, and suicide risks than Native Hawaiian Pacific Islander, and White/ European American gender minority individuals. DISCUSSION/SIGNIFICANCE: In Hawaii, one person dies by suicide every two days; suicide is the lead cause of fatal injuries. Study findings can inform future methodology studies to identify gender minorities and develop culturally relevant gender affirming mental health programs for gender minorities.

Objectives/Goals: We aimed to discover treatment candidates for uterine fibroids, a common benign tumor with adverse impacts on quality of life. Repurposing already approved medications for fibroids can expedite treatment option expansion. Using genetic proxies, we identified novel fibroid drug candidates and estimated their effect on risk of fibroid diagnosis. Methods/Study Population: We performed a genetically predicted gene expression (GPGE) analysis using S-PrediXcan and GTEx tissue models with multi-ancestry genome-wide association study (GWAS) summary statistics of fibroids (cases  =  74,294, controls  =  465). There were 81 genes significantly associated with fibroid risk. Querying drug–gene interaction databases identified 56 approved medications that target these genes, including two antihypertensives, hydralazine, and spironolactone. Using independent multi-ancestry GWAS summary statistics (N  =  635,969) for systolic (SBP) and diastolic blood pressure (DBP), we conducted GPGE analyses. Blood pressure (exposure) and fibroids (outcome) GPGE summary statistics in the same tissues were used for two-sample Mendelian randomization (MR) analyses to proxy medication effects. Results/Anticipated Results: GPGE analyses identified hydralazine/tumor protein P53 (TP53) activity and spironolactone/thyroid hormone receptor beta (THRB) activity as drug-gene candidate pairs. Both drugs increase gene activity of their paired gene. Increased TP53 expression was associated with SBP in four tissues (exposure). The MR results indicated hydralazine use, proxied by increased TP53 expression, may reduce fibroid risk by 42% per standard deviation of gene expression (odds ratio [OR]  =  0.58, p  =  1.43E-12). Increased THRB expression was associated with DBP in eight tissues and were included in the MR (exposure). The MR results suggest spironolactone use, proxied by increased THRB expression, may reduce fibroid risk by 23% per standard deviation of gene expression (OR  =  0.77, p  = 5.94E-6). Discussion/Significance of Impact: We provide biologically plausible evidence for repurposing hydralazine and spironolactone for reducing risk of fibroid diagnosis. Repurposing these hypertension medications could provide novel preventative treatments for fibroids, particularly for individuals disproportionately affected by both conditions.

Objectives/Goals: This systematic review aims to identify and synthesize evidence on the barriers and facilitators impacting the implementation of transcranial magnetic stimulation (TMS) in clinical practice, enhancing understanding for improved adoption and efficacy. Methods/Study Population: This systematic review follows PRISMA guidelines to identify relevant literature on barriers and facilitators to TMS in North America. We conducted a comprehensive search of databases including PubMed, Scopus, and PsycINFO, targeting studies published from 2000 onward. Eligible studies include qualitative and quantitative research focusing on adults aged 18 years and older in the USA and Canada. Two independent reviewers screened titles, abstracts, and full texts, extracting data on barriers and facilitators related to TMS implementation. Results/Anticipated Results: We anticipate identifying a diverse range of barriers and facilitators related to TMS implementation in North America. Expected barriers may include limited clinician knowledge, patient resistance, and logistical challenges in clinical settings. Facilitators could encompass supportive institutional policies, clinician training, and positive patient outcomes. The synthesis of findings will highlight key themes, guiding future research and practice. We aim to produce actionable recommendations for stakeholders, ultimately enhancing the effective integration of TMS in clinical care for adult populations. Discussion/Significance of Impact: This review will provide crucial insights into the barriers and facilitators of TMS implementation, informing clinicians, policymakers, and researchers. By highlighting actionable strategies, it aims to enhance TMS accessibility and efficacy, ultimately improving patient outcomes and advancing neurotherapeutic practices in North America.

OBJECTIVES/GOALS: The objective of this research is to begin documenting barriers to regenerative care to proactively address issues of accessibility and determine and implement interventions in anticipation of equitable care. We will explore the accessibility of Platelet Rich Plasma (PRP) for the treatment of knee osteoarthritis (Knee OA) METHODS/STUDY POPULATION: This research has two components. First, retrospective chart analysis using electronic databases to gather factors such as sex, race, ethnicity, disease severity, price, insurance, and treatments. Second. Prospective survey to gather the information (income, employment, education level) that are not found in the chart. Patients with knee OA conditions who received standard care will be compared to knee OA patients who received the novel PRP treatment. Statistical prediction modeling will be employed to determine the probability of receiving novel regenerative PRP treatment based on gathered explanatory variables. Our sample size is about 1000 patients per group (PRP group vs standard care group). Patients are from the three Mayo sites (Rochester, Arizona, Florida) as well as Detroit Medical Center. RESULTS/ANTICIPATED RESULTS: Our preliminary analysis shows that only a homogenous of patients (white) are receiving the novel PRP regenerative care to treat knee OA even though African Americans suffer from knee OA at a much greater rate. However, we still don’t know what is the main driver of this homogeneity. We anticipate that insurance coverage and out-of-pocket cost of care for PRP to be a huge barrier. Also, we are anticipating the lack of knowledge about PRP and its level 1 meta-analysis efficacy, as well as physician bias for not recommending PRP to certain underrepresented populations could play a role as well. We plan to order the level of influence of each barrier for each sex, race, and ethnicity, so intervention can be personalized and targeted for each population of interest. DISCUSSION/SIGNIFICANCE: Almost half of Americans live with at least one chronic medical condition and regenerative medicine holds tremendous promise to address many disease categories. To design any intervention to increase access requires knowledge about barriers in regenerative care, which is why this research is very critical now before the wide implementation of care.

OBJECTIVES/GOALS: In the crowded environment of the intestine, selected commensal bacteria and enteric viruses interact. The biological significance of this interaction, in either normal or pathological condition is not known. To study this interaction, we are developing a physiologically relevant model of an human intestinal epithelium. METHODS/STUDY POPULATION: Intestinal biopsies (ileum region) and fecal samples of 6 healthy and 6 active Crohn’s patients are being collected to derive human intestinal enteroid (HIE) lines. 2D-polarized HIE will be first characterized with studies of epithelial permeability, tight junctions and cell type composition, and co-cultured with matching fecal samples. The (co-)cultures will be then infected with human norovirus (HNoV), our model enteric virus, and infection will be quantified by RT-qPCR. In addition, the interaction of HNoV with bacteria derived from healthy or Crohn’s will be determined quantitatively by flow cytometry (viral tagging) and qualitatively by 16S sequencing of the total versus HNoV-bound bacterial species. RESULTS/ANTICIPATED RESULTS: Crohn’s patients are characterized by a microbiome dysbiosis and, in particular, by a high abundance of Enterobacteriacae. HNoV interacts with Enterobacter cloacae, and interestingly, HNoV infection is associated with exacerbation and reactivation of Crohn’s disease. By re-creating the intestinal milieu of healthy and Crohn’s patients, we expect that the kinetics of infection by HNoV will be higher in Crohn’s as compared to healthy volunteers. In addition, by studying the composition of the HNoV-bound bacterial component of Crohn’s versus heathy volunteers, we will be able to identify the contribution of selected bacteria to the expected increase of infection. DISCUSSION/SIGNIFICANCE OF IMPACT: With this study, we will fill the gap of knowledge on the importance of commensal bacteria and enteric virus interactions in healthy and diseased condition. This new knowledge will be paramount for the identification of novel strategies to combat highly prevalent virus infections.

OBJECTIVES/SPECIFIC AIMS: Due to scope and breadth of research activity and infrastructure capacities at academic medical centers, the discipline of Biomedical Informatics is often deployed in a decentralized manner through geographically dispersed and unrelated organizational units. As a result, without a conscious strategy, an academic medical center risks redundant effort and gaps in resources, and perhaps poor coordination. A mechanism to bring together disparate organizational entities to identify, discuss, and negotiate Informatics-related concerns may produce a better institutional research environment. The University of Rochester (UR) has implemented such a strategy of Informatics governance, adapting tactics from team science, diplomacy, and deliberative engagement. METHODS/STUDY POPULATION: Based on current needs and institutional Informatics priorities, the UR’s Clinical and Translational Science Institute (CTSI) established 6 Informatics “clusters” in distinct but deliberately overlapping focal areas: (1) Data—capture, management, and analysis of all types of data for research. (2) Analytics—quantitative research across the spectrum of translational research. (3) Infrastructure—technical and computing infrastructure to support informatics. (4) Electronic health records (EHR)—(i) features within the EHR explicitly designed to address the needs of research; (ii) accessing and procuring EHR data for research. (5) Population health—Informatics design and systems expertise relevant to population health research (a key CTSI focus area). (6) Education—development, deployment, and assessment of Informatics learning opportunities for learners at all levels. Each cluster facilitates access to expertise and resources around the institution, promotes collaboration, identifies redundancy, and serves as a forum to strategize regarding institutional needs related to Biomedical Informatics. A CTSI faculty or staff member leads each cluster. To maximize effectiveness of the cluster, other members are decision-makers in the organizations they represent, or serve in a critical staff function. Clusters meet in person on a quarterly basis with more frequent electronic interaction. The clusters share documents via Box, a secure online file sharing app. The cluster coordinators meet as a group on a biweekly basis to monitor progress and make plans. RESULTS/ANTICIPATED RESULTS: There were 45 different people representing 46 distinct centers, departments or offices, and 2 outside agencies agreed to participate in the clusters. In total, 20 people represented a single organizational unit; 15 represented 2 units; 8 represented 3 units, and 2 represented 4 units. The richness and complexity of these organizational linkages illustrates the decentralized nature of Informatics at the institution and the promise of the cluster approach. DISCUSSION/SIGNIFICANCE OF IMPACT: Adapting to a decentralized Informatics environment, the CTSI established clusters that recognize and respect autonomy and capacity of a wide range of units throughout the university, creating a collaborative atmosphere for steering and implementing an overall Informatics vision. As Informatics capacity rapidly expands throughout growing biomedical research institutions without a centralized Informatics hub, this distributed, deliberative approach could offer an effective governance solution that promotes cooperation. In this model, the CTSI provides the leadership and staffing necessary to ensure progress at the institutional level around Informatics and creates a venue for communication and coordination on Informatics-related topics.

OBJECTIVES/SPECIFIC AIMS: Overall Research Aim: To develop an iNKT-cell engaging reagent (“CAb”)to induce neuroblastoma-directed cytotoxicity in vitro and in a mouse model of neuroblastoma. Objective 1: Explore the contribution of different GD2 affinities to the cytotoxicity against neuroblastoma cells in vitro. Objective 2: Deteremine whether use of different stimulatory glycolipids (alpha-GalCer vs. C34) alter the activation and cytotoxicity of iNKT cells against neuroblastoma in vitro. Objective 3: To analyze survival of an immunocompetent mouse model of neuroblastoma treated with C34-loaded vs alpha-GalCer-loaded CAb molecule, and to analyze the tumor microenvironment in each treatment condition. METHODS/STUDY POPULATION: CAb molecule will be generated by fusing a CD1d protein to an scFv domain for GD2 using cloning techniques. Previous work by our group has used a streptavidin-biotin system to link CD1d to an antibody against GD2, which is large and immunogenic. Protein expression of this novel fusion protein will occur in HEK293 cells. This new CAb molecule will then be loaded with alpha-GalCer or C34 for use in cytotoxicity and in vivo experiments. Cytotoxicity Assessment: Chromium assays will be used to assess the specific cytotoxicity generated by iNKT cells against neuroblastoma cells in vitro. iNKT cells will be activated by “CAb’s” with relatively high and low affinity for GD2, and also with Alpha-GalCer and C34 glycolipid antigen. flow cytometry will be used to assess for CD107a and Interferon Gamma. Mouse Model of Neuroblastoma: TH-MYCN +/+ mice will be used as an immunocompetent model of neuroblastoma. These mice have the MYCN gene under the control of a tyrosine hydroxylase promoter, and spontaneously develop neuroblastomas by 2 weeks of life which are uniformly fatal by 8 weeks of life. In vivo survival studies will be conducted by injecting CAb of relatively high and low affinity, loaded with glycolipid antigen intraperitonealy into TH-MYCN+/+ mice starting at 2 weeks of age, twice weekly. There will also be a matched negative control. Treatment groups are listed below: 1. alpha-GalCer loaded high-affinity Cab 2. alpha-GalCer loaded low-affinity Cab 3. C34-loaded high-affinity Cab 4. C34-loaded low-affinity Cab 5. Unloaded high-affinity Cab 6. Unloaded low-affinity Cab Enrollment will be 6 mice per group for the survival curves. Tumor Microenvironment analysis: 2 additional mice will be included in each group listed above to be sacrificed 2 weeks into treatment for tumor assessment with flow cytomtetry for iNKT cell, NK cell, T-Lymphocyte frequencies as well as interferon-Gamma expression. RESULTS/ANTICIPATED RESULTS: Objective 1: We expect to find that the highest affinity scFv domains for GD2 result in the greatest amount of cytotoxicity against neuroblastoma cells via iNKT cells. Objective 2: We expect that the C34 molecule will induce the greatest amounts of iNKT cell activation against neuroblastoma cells and higher cytotoxicity against neuroblastoma, which has not been shown previously. Objective 3: We expect to see prolonged survival of mice treated with the high affinity GD2 CAb loaded with C34 or alpha GalCer compared with the low affinity CAb loaded with C34 or alpha GalCer. We also expect that the C34 loaded CAb in both groups will have prolonged survival when compared with the alpha-GalCer loaded CAbs of either affinity. DISCUSSION/SIGNIFICANCE OF IMPACT: iNKT cells have been shown previously to confer an improved prognosis in neuroblastoma and other malignancies. Furthermore, high risk neuroblastomas tend to downregulate expression of a chemokine that attracts iNKT’s to the site of the neuroblastoma. Directing iNKT to the site of neuroblastoma holds promise as an effective immunotherapy option. Our preliminary data demonstrate that CAbs directed against GD2 are capable of exerting cytotoxicity of neuroblastoma in vitro. Furthermore a trend towards prolonged survival has been shown in TH-MYCN mice in early experiments. The development of a novel antibody that has reduced immunogenicity, incorporates a glycolipid antigen that does not induce iNKT cell anergy, and is specific for the GD2 tumor specific antigen has potential to result in increased iNKT-mediate neuroblastoma cytotoxicity and prolonged survival in TH-MYCN+/+ mice.

OBJECTIVES/GOALS: Patient recruitment, enrollment, and retention continues to be one of the leading barriers to successful clinical trials, and results do not always reflect the diversity of the general population. This systematic review aims to assess the impact of decentralized methods on recruitment, retention, and diversity on recent clinical research. METHODS/STUDY POPULATION: A systematic search of the literature, using databases such as PubMed, Cochrane Library and EMBASE to find publications reporting on the aspect of recruitment in decentralized clinical trials was performed. The titles and abstracts of the publications were assessed, excluded those lacking sufficient or pertinent information regarding decentralization in clinical trials. The remaining publications were reviewed for those reporting sufficient data regarding the impact of decentralization on the aspect of recruitment in clinical trials to be included in the focused analysis. Studies reporting on participant retention and diversity in addition to recruitment were emphasized. RESULTS/ANTICIPATED RESULTS: This systematic search returned 13 studies highlighting the role of decentralized clinical trial methods impacting participant recruitment, retention, and diversity in clinical trials. Out of the 13 studies, 10 reported improved recruitment using virtual or decentralized methods, and 7 of these reported improvements when compared alongside with traditional methods. 7 studies reported a positive impact on participant retention, with 4 of these directly comparing decentralized methods with traditional methods. Lastly, of these studies, 5 were reported to have trended towards diversity in the demographics of the sample population, including race or geographic location. DISCUSSION/SIGNIFICANCE: Related reviews have stated a lack of published comparable data to determine if DCTs (Decentralized Clinical Trials) improved recruitment and retention. Results suggest this review addresses such a gap, by providing data on how decentralized methods can benefit recruitment and retention, potentially highlighting a new standard.

OBJECTIVES/SPECIFIC AIMS: The goal of this study was to determine if there are any sex differences in the pathophysiological effects of sFR. METHODS/STUDY POPULATION: Male Fischer rats (4-month-old) were maintained on a control (CT) (ad libitum regular chow; n=8) or sFR (60% reduction of daily food intake, n=8) diet for 2 weeks. On days 1, 2, 3 and 14, the rats were placed in metabolic cages for food and water intake and 24-hour urine collection. Body weight (BW) is measured daily. After 2 weeks, the animals are given free access to normal chow for 3 months. Short-term and long-term effects of sFR on blood pressure and heart rate will be measured. RESULTS/ANTICIPATED RESULTS: After 2 weeks, the male CT group gained 7% BW (p <0.05), while BW in the sFR males was reduced by 12% (p<0.05 vs. CT). In contrast, female controls did not gain BW while the sFR females lost 18% of their BW. Water intake was reduced by 35%, which was similar to the reduction in females (p=0.18). The hematocrit of sFR male rats was higher (51.1%) than the CT group (45.2%, p<0.05), which was most likely due to the 6% reduction in plasma volume. A similar effect on hematocrit was observed in sRF females. Similarly, also to female rats, sFR had no effect on Na+ and K+ plasma or urine concentrations by day 14 in the male rats. DISCUSSION/SIGNIFICANCE OF IMPACT: sFR has similar effects on electrolyte balance in males and females. Ongoing studies will determine if there is any sex difference in the effects of sFR on blood pressure, heart rate and susceptibility to hypertension and cardiac injury.

OBJECTIVES/GOALS: 1) Investigate the network level interactions of attention and learning during an attention network task (ANT) and an implicit learning contextual cueing (CC) task. 2) Assess the effect attention rehabilitation strategies have on behavioral and neural responses pre/post-attentional intervention. METHODS/STUDY POPULATION: This study involves refractory epilepsy patients with implanted intracranial electrodes and moderate-to-severe traumatic brain injury (m/sTBI) survivors. In epileptic patients, we will identify connectivity of cortical regions via the ANT, which probes components of attention (alerting, orienting, and executive control) and a CC task that probes implicit learning. We hypothesize that modulation of attention and learning can be seen at the network level. In TBI we will assess improvement following two attention rehabilitation paradigms behaviorally; and use our results from epileptic patients to guide measurement of treatment-related neuroplastic change via scalp electroencephalography. RESULTS/ANTICIPATED RESULTS: When the proposed objectives are complete, we expect to determine how the implicit learning rate in m/sTBI changes as a result of both direct attention and metacognitive-strategy training, and discern the neuroanatomical networks associated with attention and implicit learning based on connectivity results. We expect to identify intracranial regions and networks that exhibit modulatory effects associated with attention and implicit learning. Additionally, we anticipate that deficits in attention will be mitigated following training and hypothesize that implicit learning rate will improve in TBI patients as a result of both attentional rehabilitation paradigms. DISCUSSION/SIGNIFICANCE OF IMPACT: Characterizing intracranial activity in epilepsy patients will give electrophysiology data unattainable in TBI patients. This intracranial perspective will enable us to propose mechanisms of action that may result from our interventions and enable critique of current rehabilitation treatments.

OBJECTIVES/GOALS: Venous thromboembolism (VTE) is a major cause of morbidity and mortality. Due to its relatively low incidence, prospective studies are limited. This makes administrative claims a promising data source to study VTE. We sought to examine the reproducibility of results using different VTE definitions from the published literature. METHODS/STUDY POPULATION: We conducted a retrospective analysis of a random 10% sample of the 2010-2022 IQVIA LifeLink PharMetrics Plus™ database, an administrative claims database representative of the commercially insured population of the United States. We selected cancer patients undergoing major gastrointestinal surgery, who have a higher risk for postoperative VTE (deep venous thrombosis [DVT] and/or pulmonary embolism [PE]). VTE was defined using ICD-9-CM and ICD-10-CM codes using definitions from 4 individual published studies. We compared the 4 definitions with respect to the incidence of VTE and factors associated with post-discharge VTE using standard univariate and multivariable logistic regression models. The same logistic regression models were used for each of the 4 definitions. RESULTS/ANTICIPATED RESULTS: There were substantial differences in VTE coding among the 4 definitions (range 107 to 225 ICD-9/10 codes for DVT and 12 to 24 codes for PE). The eligible population comprised 2,360 patients (49% female) with a median age of 49 years (interquartile range 47-52 years). During the index surgery hospitalization, a total of 58, 62, 63, and 83 patients developed VTE using the 4 definitions. In the 2,126 patients eligible for VTE prophylaxis, a total of 108, 68, 73, and 107 patients developed post-discharge VTE (range for DVT 35 to 81, range for PE 39 to 76). On multivariable analysis, factors independently associated with VTE included age using 1 of 4 definitions, esophageal surgery type using 3 of 4 definitions, and liver surgery type and Elixhauser score using all 4 definitions. DISCUSSION/SIGNIFICANCE: The incidence of VTE is directly affected by differences in ICD-9/10 codes used. Definitions for important clinical outcomes should be standardized when using administrative claims data in order to improve reproducibility of findings.

OBJECTIVES/SPECIFIC AIMS: (1) Obtain publically available citation data, funding data, and generate multiple networks topologies based on dynamic queries of individual faculty. (2) Determine successful pathways that lead to tenure, and career advancement, in addition to determining the effect of CTSA programs on faculty collaboration. (3) Develop publically available commercial interface for the study of faculty networks METHODS/STUDY POPULATION: For our study we included all available citation and funding data publically available on all CTSA programs (as of 2015) with historical data dating back to 2005. We then included the top 25 collegiate institutions who may not have had a CTSA program (eg, Princeton University). We then developed network topologies for each university network, and explore the evolution of individuals in these networks, and the effects of faculty development—as an example in the University of Rochester network, we singled out the directors of the CTSA program there to understand their level of centrality and overall impact on network development, with key observations being that early publications across varying domains lead to stronger network performance. Although individuals who did not benefit from such development, may have succeeded but if they did were likely to leave the institution for elsewhere. RESULTS/ANTICIPATED RESULTS: A secondary goal of this project is to evaluate the effectiveness of the Clinical & Translational Science Institute (CTSI) since its inception in 2006. The mission of CTSI is to advance the field of translational science and research, to link other departments at URMC and community stakeholders by research collaboration, publication, and goals to improve population health, and provide translational education and training to students, researchers, and physicians. To determine how the induction of CTSI affects collaboration within the URMC network, we examined the role of funding in the CTSI network. This was done around the second successful funding around 2013. In doing so we can see that not only did the funding request affect the network topology, but opened new collaborations which were not present prior to the request. DISCUSSION/SIGNIFICANCE OF IMPACT: We have developed an automated method, which is superior to manual methods necessary for citation generation and funding data analysis of faculty growth in citation networks. This technique is applicable to all institutions, not just those in a CTSA environment, but demonstrates the benefit of cross-collaborative efforts, in the case of the URMC network we can state the following. The key takeaway is for individuals to succeed in the URMC collaborative environment they should create their own network and expand it and eventually rise to prominence. There are 2 pathways to this you can take the Dewhurst approach which is to seek out collaborations among internal peers and scale up. Or you can take the Nedergaard approach which is develop the special network, and gain enough public recognition outside of the network that you are capable of leaving it (Fig. 2d). In either case, collaborations among communities and diverse out-degree networks allow faculty to succeed in their given field. Given the wealth of data which has been curated in this fashion, there are numerous explicit questions that can be asked of the data. One of the unique approaches of this data is that is highly reproducible, which allows various questions to be asked. Future work would try to determine what optimal pathways are in a given network to success, and who are ideal collaborators, and collaborations to avoid. Given this information, custom pathways to career success for individual faculty can be developed, moving beyond purely institutional level co-citation networks, which do little to advance faculty development at scale. In Figs 1c and d, the network increased by 75% in terms of graph density (0.007) and decreased by 18.8% (16) in terms of diameter. What this suggest in that the interconnectivity of the network grew dramatically, while the ability for new members to integrate into it increased. This also apparent when one examines the modularity of the network down by 3.6% (0.857), this suggest that the network has as many communities but these communities are less isolated that those in the previous funding year, meaning fields are becoming more transdisciplinary in their collaborations. This was the result of the presence of a CTSA program, thus demonstrating the effectiveness of such institutions, however, our analysis also lays the framework for applying this to other institutions which may be considering a CTSA. Or maintaining the success of a given CTSA program, and ultimately determining where faculty should place their efforts and choose which programs to pursue career advancement.