OBJECTIVES/GOALS: Our overall objective is to investigate the relationship between radiologic features of meningioma with recently identified histopathological and molecular biomarkers, and to apply a machine learning (ML) approach to further demonstrate their utility in predicting clinical outcomes. METHODS/STUDY POPULATION: We have enrolled a cohort of 84 patients with meningioma diagnosed on the basis of conventional gadolinium-enhanced MRI imaging features since September 2019. Each patient has demographic and clinical data, Ga-68-DOTATATE MRI/PET SUV and dynamic metrics, DCE-MRI perfusion parameters, and histopathologic data. Various tumor subregions will be segmented semi-automatically and later confirmed by experienced neuroradiologist. Histopathologic data will include histologic grade, mitotic rate, Ki67 proliferative index, and presence of WHO established atypical histologic features, immunohistochemical parameters, and established high-grade molecular features. We will use supervised learning techniques to develop algorithms for predicting molecular features from imaging phenotypes. RESULTS/ANTICIPATED RESULTS: Anticipated results - advancements in understanding the molecular biomarkers of meningiomas has uncovered genetic alterations and epigenetic changes that more accurately determine tumor behavior. Currently, the imaging correlates of these molecular biomarkers are unknown, and utilizing radiographic data to predict prognosis and imaging-based classifications of meningiomas have not yet been investigated. Validated imaging correlates of molecular biomarkers not only provide an in-vivo assessment of tumor biology, but can also be integrated with histopathologic features ( radiopathomics models’) for more accurate disease prognostication. We anticipate that our results will identify surrogate imaging features for some of the recently emerged molecular biomarkers of meningioma. DISCUSSION/SIGNIFICANCE: There is a paucity of data on the importance of imaging phenotypes in determining tumor biology. This work has the potential of significant clinical impact by enabling a priori molecular characterization of meningiomas at the time of new diagnosis or recurrence, thereby allowing a personalized medicine approach to treatment planning.

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.

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: 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: 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.

OBJECTIVES/GOALS: Our long term goal is to identify the socioeconomic and cancer-specific determinants in patients with prostate cancer undergoing prostatectomy that impact their ability to maintain a healthy weight. This study explores the association between participants’health determinants and their indicated degree of self-efficacy and quality of life (QoL). METHODS/STUDY POPULATION: Study population includes participants of the WARRIOR trial (n=40), which included overweight men scheduled radical prostatectomy from the University of Kansas Medical Center. In addition to baseline demographics, the study team will administer a questionnaire based on a socioeconomic position and health-related QoL framework. This questionnaire will assess participants' socioeconomic, cancer-specific, and psychological circumstances at time of surgery and present day. Univariate analyses will be conducted on all variables with bivariate analyses between socioeconomic and clinical items to the outcome of composite self-efficacy scoring. RESULTS/ANTICIPATED RESULTS: We anticipate that participants in the intervention will report higher self-efficacy and emotional/social support than participants in the control group, participants with social vulnerability (lower income, marginalized race/ethnicity, etc) will report decreased self efficacy and poorer QoL compared with participants who are not socially vulnerable, and that participants who previously indicated social vulnerability will report more emotional barriers to weight loss, and lesser weight loss satisfaction, self-esteem, and QoL. DISCUSSION/SIGNIFICANCE: Lifestyle interventions have helped prostate cancer patients lose weight before surgery, but many regain weight. Exploring perceptions of self efficacy as well as learning more about what structural and systemic barriers affect self efficacy is important to inform how to improve our approach for sustained weight loss and health behavior changes.

OBJECTIVES/GOALS: Sleep is critical for healing, however pediatric intensive care unit (PICU) sound is above recommended levels (i.e., 45 A-weighted decibels [dBA]). This observational study identifies sources of PICU sound and compares sources between times of high (i.e., dBA≥45) and low (i.e., dBA < 45) levels. METHODS/STUDY POPULATION: The sound environment of 10 critically ill children 1 to 4 years of age was monitored via a bedside dosimeter and video camera for 48 hours, or until PICU discharge. Dosimeter and video data were uploaded to Noldus Observer XT and time synchronized. A reliable, previously published coding scheme developed to identify sound sources in the adult ICU was modified for the pediatric population. Sound sources (e.g., clinician/family/child [verbal vs. non-verbal] vocalization, patient care, medical equipment) were identified via instantaneous sampling of video data at each minute of recording. The proportion of sampling points with each sound source are compared between times of high and low sound levels, and between day (7:00-18:59) and night (19:00-6:59) shift. RESULTS/ANTICIPATED RESULTS: Video coding is ongoing, with high inter-rater reliability (κ̅=0.99, SD DISCUSSION/SIGNIFICANCE: Medical equipment sound is ubiquitous in the PICU. Clinicians should optimize the PICU sound environment for sleep, including minimizing equipment alarms, conversation, general activity, and screen media during child rest. Large-scale studies are needed to confirm findings from this small cohort.

OBJECTIVES/GOALS: Treatment options for glioblastoma (GBM) are limited. Prognosis remains dismal, with an 18 month on average survival rate following diagnosis due to treatment resistance and disease recurrence. The goal of this project is to investigate hallmarks of cancer progression that contribute to temozolomide (TMZ) resistance, a first tine treatment for GBM. METHODS/STUDY POPULATION: Two signaling pathways were investigated in TMZ-sensitive and -resistant GBM cell lines and in primary and recurrent patient-derived xenograft (PDX) tumor cells by genetically and pharmacologically inhibiting methionine adenosyltransferase 2A (MAT2A) and adenosylhomocysteinase (AHCY). Cell growth and survival were assessed by measuring protein expression of proliferation, oxidative stress and cell cycle arrest markers. EPIC array analysis and targeted bisulfite sequencing were conducted to identify changes in genome-wide and specific CpG island methylation. The Seahorse XF Analyzer measured mitochondrial respiratory capacity and oxidative metabolism. Induced pluripotent stem cell organoids were co-cultured with PDX tumor cells to determine if treatments mitigate tumor cell invasiveness. RESULTS/ANTICIPATED RESULTS: Compared to parental cells (PC), MAT2A gene expression was increased by 1.7-fold in acquired resistant and de novo resistant GBM cells (RC) [(transcript per million): PC, 7386 ± 0.012; RC, 12925 ± 0.023; n=2; p=2.10e-8]. Compared to TMZ-sensitive cells (TS), TMZ-resistant cells (TR) demonstrated a 56% increase in baseline oxygen consumption rate [(pmol/min): TS, 179 ± 6.7; TR, 279 ± 13; n=18; p=.0012] and 64% increase in maximal respiratory capacity [(pmol/min): TS, 403 ± 29; TR, 659 ± 35; n=6; p DISCUSSION/SIGNIFICANCE: MAT2A and AHCY contribute to TMZ resistance and recurrence by dysregulating methylation programs and upregulating antioxidant programs, respectively. These findings provide a foundation for developing novel combinatory therapeutic strategies and inform clinical studies intended to increase remission and reduce recurrence for GBM patients.

OBJECTIVES/GOALS: Supported by the State of Alabama, the Alabama Genomic Health Initiative (AGHI) is aimed at preventing and treating common conditions with a genetic basis. This joint UAB Medicine-HudsonAlpha Institute for Biotechnology effort provides genomic testing, interpretation, and counseling free of charge to residents in each of Alabama’s 67 counties. METHODS/STUDY POPULATION: Launched in 2017, as a state-wide population cohort, AGHI (1.0) enrolled 6,331 Alabamians and returned individual risk of disease(s) related to the ACMG SF v2.0 medically actionable genes. In 2021, the cohort was expanded to include a primary care cohort. AGHI (2.0) has enrolled 750 primary care patients, returning individual risk of disease(s) related to the ACMG SF v3.1 gene list and pre-emptive pharmacogenetics (PGx) to guide medication therapy. Genotyping is done on the Illumina Global Diversity Array with Sanger sequencing to confirm likely pathogenic / pathogenic variants in medically actionable genes and CYP2D6 copy number variants using Taqman assays, resulting in a CLIA-grade report. Disease risk results are returned by genetic counselors and Pharmacogenetics results are returned by Pharmacists. RESULTS/ANTICIPATED RESULTS: We have engaged a statewide community (>7000 participants), returning 94 disease risk genetic reports and 500 PGx reports. Disease risk reports include increased predisposition to cancers (n=38), cardiac diseases (n=33), metabolic (n=12), other (n=11). 100% of participants harbor an actionable PGx variant, 70% are on medication with PGx guidance, 48% harbor PGx variants and are taking medications affected. In 10% of participants, pharmacists sent an active alert to the provider to consider/ recommend alternative medication. Most commonly impacted medications included antidepressants, NSAIDS, proton-pump inhibitors and tramadol. To enable the EMR integration of genomic information, we have developed an automated transfer of reports into the EMR with Genetics Reports and PGx reports viewable in Cerner. DISCUSSION/SIGNIFICANCE: We share our experience on pre-emptive implementation of genetic risk and pharmacogenetic actionability at a population and clinic level. Both patients and providers are actively engaged, providing feedback to refine the return of results. Real time alerts with guidance at the time of prescription are needed to ensure future actionability and value.

OBJECTIVES/GOALS: Nearly all thoracic aortic aneurysm patients suffer from hypertension leading to elevated wall tension and abnormal extracellular matrix remodeling. PTSD patients have higher blood pressure both at rest and in response to stimuli. Although stress is associated with cardiovascular disease, the exact mechanism linking the two is still unknown. METHODS/STUDY POPULATION: Adult C57BL/6 mice underwent a PTSD induction protocol consisting of inescapable foot shock followed by single prolonged stress. The mice were assessed incrementally for their PTSD-like phenotype using specific behavioral tests chosen to assess for each of the human criteria of PTSD according to the DSM-V. Tail cuff blood pressure measurements were taken serially throughout the 16-week protocol. At terminal study, thoracic aortic diameter measurements were obtained through digital microscopy and plasma was harvested for cytokine analysis. Thoracic aortic aneurysms (TAA) were induced through periadventitial application of a calcium chloride solution on the descending thoracic aorta in BPH/2J and BPN/3J adult mice. The thoracic aortic diameter was measured at terminal study through digital microscopy. RESULTS/ANTICIPATED RESULTS: Using our PTSD-like mouse model we have demonstrated that PTSD-like mice have significantly higher systolic blood pressure following a reminder of the traumatic event than control mice recapitulating the human phenotype. They also had increased plasma proinflammatory cytokines and larger thoracic aortic diameters than control mice. Although the increased thoracic aortic diameter is not an aneurysm, it suggests ECM remodeling is occurring predisposing the aorta to aneurysm formation. Finally, we have shown that in neurogenic hypertensive mice, TAA formation was accelerated by 12 weeks with roughly 70% dilation at 4 weeks post-TAA induction surgery as compared to roughly a 20% dilation in control mice. DISCUSSION/SIGNIFICANCE: Altogether, these studies reinforce the link between stress and TAA development, and our mouse model will allow for the underlying mechanism to be elucidated. Better understanding of the mechanism linking PTSD and TAA will allow for the creation of novel therapeutics to treat PTSD symptoms while also delaying TAA progression.

OBJECTIVES/GOALS: A growing number of older adults in the United States have multiple social determinants of health (SDoH) that are barriers to effective medical care. We used generalizable machine learning methods to identify and visualize subtypes based on participant-reported SDoH profiles, and their association with delayed medical care (self-reported yes/no). METHODS/STUDY POPULATION: Data. All participants aged >=65 in All of Us with complete data on 18 SDoH self-reported variables, selected through consensus by 2 experienced health services researchers, and guided by Andersen’s behavioral model. Covariates included demographics, and the outcome was delayed medical care . Cases (n=4090) consisted of participants with at least one of the 18 SDoH variables, and controls (n=7414) consisted of participants with none of them. Method. (1) Used bipartite network analysis and modularity maximization to identify participant-SDoH biclusters, and visualize them through ExplodeLayout. (2) Used multivariable logistic regression (adjusted for demographics and corrected through Bonferroni) to measure the odds ratio (OR) of each participant bicluster to the outcome, compared with the controls. RESULTS/ANTICIPATED RESULTS: The analysis identified 7 SDoH subtypes (https://postimg.cc/Vd7Pg4xZ) with statistically significant modularity compared with 100 random permutations of the data (All of Us=.51, Random Mean=.38, z=20, P DISCUSSION/SIGNIFICANCE: The results identified 7 distinct subtypes based on SDoH profiles and their risk for delayed medical care, highlighting the importance of addressing specific combinations of barriers, with affordability having the highest risk. Furthermore, the analytical methods used are generalizable and have been made publicly available on CRAN and All of Us.

OBJECTIVES/GOALS: Targeting the V-domain immunoglobulin suppressor of T cell activation (VISTA) signaling pathway has been suggested as a promising approach for overcoming resistance to current immune checkpoint therapies in advanced cancer. This review will synthesize the rapidly-expanding literature on VISTA protein expression on prognosis in various cancers. METHODS/STUDY POPULATION: To determine the prognostic significance of high VISTA expression across treatment-naïve malignant tumors, a systematic review and meta-analysis will be performed of published cohort studies which measured VISTA protein expression on solid tumors. Primary and secondary outcome endpoints of overall survival (OS) and disease-specific survival (DSS) will be compared across cohort studies using a random effects model to calculate pooled hazard ratios (HRs) for each time-to-event end point with 95% CIs. For articles that only provide Kaplan-Meier (KM) curves, the Engauge Digitizer software will be used to measure the time and survival probability coordinates on the KM curves to estimate the HRs. Correlations of VISTA expression and clinicopathological characteristics will be evaluated by pooled risk ratios. RESULTS/ANTICIPATED RESULTS: A search of 4 electronic databases including Pubmed, Embase, Web of Science and Cochrane resulted in 5578 publications of which 66 containing a broad spectrum of malignant solid tumors will undergo full-text review for study inclusion. Tumor types most represented with at least 3 articles include lung, pancreas, skin, head & neck, colorectal, mesothelioma, cervix, soft tissue, breast, liver and ovarian. Our working hypothesis is that the pooled HR for high VISTA expression on overall survival will be approaching 1.0 given conflicting reports across the cancer literature. Risk of bias will be assessed across studies. Quantifications of heterogeneity will be assessed by visual exploration of forest plots as well as by multiple statistical metrics including the Q statistic and the I²coefficient. DISCUSSION/SIGNIFICANCE: The results of this systematic review and meta-analysis will provide a more comprehensive understanding of VISTA’s prognostic role both across all malignant tumors and for subgroups of similar tumor types which may impact the types of tumors and tumor microenvironments selected for early trials of anti-VISTA therapy.

OBJECTIVES/GOALS: Adult spinal deformity is commonly treated by spine surgeons. Patient treatment planning and surgical candidacy are dependent on static measurements and inconsistent heuristics which lead to high complication rates and poor outcomes. This study tests the role of supplemental longitudinal and dynamic patient data in improving surgical planning. METHODS/STUDY POPULATION: Ten adult spinal deformity surgeons at Johns Hopkins Hospital were interviewed for 30 minutes by the study team. The script was reviewed by the institutional review board to alleviate any risk of bias. Two patient sets were curated utilizing previously treated, anonymized patient data sets from a non-surveyed practitioner. Each patient set was coupled with relevant radiographic imaging (MRIs, CTs, and plain radiographs) and pertinent clinical information that is collected in a standard clinic visit. Surgeons were presented with a patient and asked to note their specific surgical plan. Subsequently, surgeons were presented with four sets of supplemental dynamic spine data and asked to note their surgical plan for each set. Shaprio-Wilks and Mann-Whitney U tests were used to assess data normality and nonnormality. RESULTS/ANTICIPATED RESULTS: Preliminary data has shown inconsistency in both surgical selection and surgical type amongst physicians when presented with initial clinical findings and radiographic reports for base patient cases. There was minimal consensus among surgeons on the number of levels fused and interbody spacer usage. Early results show that dynamic spine data may be beneficial in creating consistency between surgeons, despite inter-surgeon variability in surgical planning without this data. Posture, pain location, pain severity, and quantified activity throughout the day have been referenced as the most useful dynamic spine data to consider. Amongst all providers, the availability of dynamic spine data resulted in a change in surgical planning. DISCUSSION/SIGNIFICANCE: Recent publications have shown that spine surgery patient candidacy and surgical planning are dependent on heuristics. This has led to inconsistencies amongst surgeon preferences and increases in improper patient selection for procedures. Incorporating longitudinal dynamic data may lead to increased consistency and improved patient outcomes.

OBJECTIVES/GOALS: Anti-cancer therapies, such as chemotherapy, can induce senescence. Senescent cells may produce factors that can promote tumor progression. In this study, we will investigate the effect of senolytics and anti-cancer treatment on fibroblasts, which are a part of the tumor microenvironment, and patient-derived colorectal cancer organoids. METHODS/STUDY POPULATION: We will induce senescence in fibroblast lines via irradation. Induction of senescence will be confirmed by monitoring SASP production, changes in morphology and proliferation rates, and senescence-associated b-galactosidase activity. To investigate the efficacy of senolytics on senescence-induced fibroblasts and CRC tumor organoids, we will creat a dose response curve and calculate IC50 values for proliferating fibroblast, senescent fibroblasts and CRC organoids. To identify the synergistic effects of anti-cancer and senolytic compounds, including Navitoclax and Dasatinib, on fibroblasts and CRC organoids, we will create dose matrixes using senolytics at concentrations that were shown to have senolytic activity and drugs from an anti-cancer library. RESULTS/ANTICIPATED RESULTS: If senescence is induced in the fibroblast lines, we expect to see no changes in confluency over 4 days, the morphology will change from a thin, spindly shape to a flattened shape, and senescence-associated b-galactosidase activity will be observed. After the fibroblast lines are treated with potential senolytic compounds, we would expect to see decreased viability in the senescence-induced fibroblast lines when compared to proliferating fibroblast lines. We predict that the viability of CRC organoid lines will slightly decrease at high concentrations of the senolytic due to overall toxicity. We expect that the senolytic and anti-cancer compounds will have a synergistic effect. Senolytic activity could reduce the senescent cell population that was developed in response to anti-cancer therapy. DISCUSSION/SIGNIFICANCE: There is an increased interest in identifying compounds that selectively promote apoptosis in senescent cells. This study uses a cell-based approach to validate senolytic activity of compounds with senolytic potential in senescence-induced fibroblast lines and investigates the synergistic effects of senolytics and anti-cancer compounds on CRC.

OBJECTIVES/GOALS: Single nucleotide polymorphisms in the transcription factor 7-like 2 (TCF7L2) gene are associated with Type 2 Diabetes (T2D) and nonalcoholic fatty liver disease (NAFLD). The metabolic function of TCF7L2 in the liver remains to be fully elucidated, but we hypothesized that TCF7L2 contributes to NAFLD through regulation of zonal metabolic pathways. METHODS/STUDY POPULATION: Using single nuclei RNA sequencing, we examined Tcf7l2 expression in periportal (PP) hepatocytes around the portal triad and pericentral (PC) hepatocytes surrounding the central vein of the liver. To visualize TCF7L2 transcriptional activity we used a TCF reporter mice, which expresses an H2B-eGFP fusion protein downstream of the conserved TCF DNA binding site. We disrupted Tcf7l2 transcriptional activity in mouse liver by breeding mice with a floxed Tcf7l2 exon 11, which encodes part of the DNA binding domain (DBD), to albumin-Cre mice (Hep-TCF7L2ΔDBD). Eight-week-old mice were fed a choline-deficient amino acid-defined high fat (CDAHFD) diet for 8 weeks. In liver samples harvested from these mice, we examined disruption to several key zonated metabolic pathways, and quantified the development of fibrosis. RESULTS/ANTICIPATED RESULTS: Single nuclei analysis revealed that Tcf7l2 mRNA was expressed primarily in parenchymal cells of the liver but was ubiquitous across the liver lobule. However, in immunofluorescence analysis of TCF reporter mice, the transcriptional activity of TCF7L2 was highly restricted to PC hepatocytes. Classic PC hepatocyte markers, including glutamine synthetase (Glul), were absent in Hep-TCF7L2ΔDBD mice. Following the CDAHFD, Hep-TCF7L2ΔDBD mice developed more severe fibrosis in histological analysis, and expressed elevated levels of genes involved in fibrogenesis, collagen synthesis and TGFβsignaling. Hep-TCF7L2ΔDBD mice also displayed hepatic cholesterol accumulation following the CDAHFD, which was likely the result of impaired pericentral bile acid synthesis. DISCUSSION/SIGNIFICANCE: Our results suggest that TCF7L2 plays an important role in the regulation of zonated metabolic pathways, which may contribute to the development of fibrosis. Ongoing analyses are exploring the mechanisms regulating the zonal transcriptional activity of TCF7L2.

OBJECTIVES/GOALS: Nearly 3 million Americans live with arm impairment following stroke. While as many as 20% of patients fully recover, individual differences in recovery make one-size-fits-all rehabilitation approaches suboptimal. The goal of this study was to use our custom rehabilitation platform to identify neuromuscular biomarkers of arm control in stroke. METHODS/STUDY POPULATION: Chronic stroke survivors (N = 10) reached for targets in a virtual reality environment using both hands. They completed 162 reaches divided into 3 blocks. Following baseline, we used our custom exoskeletons to provide 50% arm weight assistance to the impaired limb and 50% arm weight resistance to the non-impaired limb. We removed the exoskeletons during the retention block. We used electromyography to approximate muscle activity in the anterior deltoids. Relative contribution (RC) was calculated as the displacement of the impaired arm divided by the sum of displacements for both arms. Muscle contribution (MC) was calculated as the root mean square of impaired arm muscle activity divided by the sum of activity for both deltoids, normalized to maximum voluntary contraction. RESULTS/ANTICIPATED RESULTS: During baseline, RC of the impaired limb was 43%; patients reached significantly less with their impaired arm compared to their non-impaired arm (p = 0.02). MC of the impaired deltoid was 56% and was similar between arms (p = 0.5). During loading, RC did not change relative to baseline (p = 0.87), but MC tended to decrease by 11% (p = 0.12). These results suggest a tradeoff between kinematic and muscular control of reaching. This new finding closely matches our previous work in 12 healthy controls, where we found a 2% increase in RC and a 11% decrease in MC. Importantly, 4/10 patients exhibited an inverse tradeoff (i.e., decrease in RC and/or increase in MC). We will analyze neuroimaging data to determine the role lesion size and location play in predicting an individual’s response to gravity compensation. DISCUSSION/SIGNIFICANCE: Our tradeoff analysis serves as a potential neuromuscular biomarker of stroke survivors’ responsiveness to gravity compensation. This forms the basis for personalized technologies for stroke rehabilitation. With further development, clinicians can use our platform to fine-tune compensation levels based on the individual needs of the patient.

OBJECTIVES/GOALS: The purpose of this pilot study is to evaluate the clinical utility and patient/provider acceptability of a buprenorphine (bup) precision dosing approach for opioid use disorder (OUD) in Puerto Rico (PR) to in estimate the most adequate bup dosing regimen based on the unique pharmacological and clinical characteristics of these patients. METHODS/STUDY POPULATION: The goal of this pilot study is to evaluate the extent to which people delivering (providers) or receiving (patients) opioid use disorder care in PR consider our 'bup precision dosing approach' to be appropriate, based on anticipated or experienced cognitive and emotional responses. We will use the Theoretical Framework of Acceptability (TFA) to conduct this evaluation. We expect to generate a baseline understanding of the acceptability of our bup precision dosing approach in terms of clinical utility and attitudes by OUD patients and providers in clinics in PR, We will conduct focus groups and surveys to document patients and providers perceptions, beliefs, attitudes and reception of our bup evidence-based dosing approach. RESULTS/ANTICIPATED RESULTS: We seek to answer the following questions: How do OUD providers and patients in PR view, and how will they engage with our buprenorphine precision dosing approach? Will our intervention based in science be accepted be these individuals? What are their attitudes towards this? How they perceive the efficacy of this intervention to be? What are the barriers and facilitators of this evidence based intervention? This knowledge is crucial before clinical implementation is pursued, we expect to comprehend the unique attitudes and perceptions of these population that supports the successful implementation in the nearby future and enhance the innovation uptake of our bup dosing model for OUD in PR. DISCUSSION/SIGNIFICANCE: It is important that adequate assessments that assess acceptability and feasibility prior to implementation and while still in developmental phases are conducted to plan ahead for the implementation of interventions since innovation uptake depends largely on contextual factors, not just innovation effectiveness.

OBJECTIVES/GOALS: There is a need for high-quality and efficient translation of health technologies in the Ontario healthcare system. The goal of this project is to understand the decision-making processes of government expert groups developing recommendations for the system-level implementation of pharmacogenomic testing. METHODS/STUDY POPULATION: This prospective observational case study includes the Ontario Health Pharmacogenomics (PGx) Working Group focused on developing recommendations for a PGx testing implementation strategy in the province. Ontario Health is the government agency that oversees provincial healthcare planning and service delivery. Using qualitative ethnographic methods, we will observe and document the working group’s activities over a 10-month period. Data collection involves meeting recordings, correspondences, researcher field notes, decision-making processes, and group characteristics. Using descriptive statistics and inductive qualitative analyses, the data will be examined to build theory and frameworks for knowledge translation. RESULTS/ANTICIPATED RESULTS: The results will be presented through a case report, process maps, decision milestones, visualizations, and procedural recommendations for future expert groups. This study will contribute to the body of foundational knowledge about translational sciences and support the National Center for Advancing Translational Sciences’guiding principles. To enhance translational processes and train the future translational workforce, this research can be used for educational initiatives. In addition, the observed processes will inform a theory about how expert recommendations are developed in public healthcare systems. DISCUSSION/SIGNIFICANCE: This research addresses a current gap in understanding around translational processes, government decision-making, and the development of recommendations for the adoption, implementation, and dissemination of the novel health technologies transforming public healthcare in Canada.

OBJECTIVES/GOALS: The overall objective of this study is to determine if medial arterial calcification (MAC) is an independent predictor of diabetes and to evaluate the relationship between MAC, periodontitis and Type II diabetes mellitus. METHODS/STUDY POPULATION: A retrospective case-control model analyzing radiographs for periodontitis and MAC to identify potential biomarkers for underlying systemic conditions, such as diabetes. Charts of patients attending UPENN School of Dental Medicine clinics between 2015 and 2022 were reviewed. Demographics, medical and dental history, diabetic status (identified by POC blood glucose level, fasting blood glucose and/or A1C), and medication history were documented amongst other variables. Patients aged 18 years or older with diabetes and having full mouth intraoral radiographs (FMX), panoramic radiographs and CBCTs were included. Patients with radiographs of poor quality were excluded. Multivariate analysis was used to determine possible associations between diabetes and periodontitis among persons with or without MAC. RESULTS/ANTICIPATED RESULTS: In our pilot study involving 28 participants, 53% of the population with moderate or severe periodontitis had MAC. By the Fisher’s Exact Test, there was an association, meaning those with more periodontal disease are more likely to have MAC (p=0.014). Sixty-three percent of patients with diabetes had MAC, while 19% of patients without evidence of diabetes also had MAC, (p=0.067). There was not enough evidence of association between diabetes and presence of MAC at this time, due to a small sample size, however there was a high prevalence of MAC among the diabetics. We hypothesized that periodontitis, a condition that shares many risk factors with diabetes would also be associated with incident MAC. Findings from this study will be key for the implementation of preventive screening protocols and referral systems. DISCUSSION/SIGNIFICANCE: Diabetes is on the rise and about half of diabetics are undiagnosed. CBCT imaging frequently used in dentistry can detect incidental findings such as MAC. This study has the potential of detecting statistically significant links between MAC, periodontitis and diabetes, hence serving as a sensitive radiographic biomarker for diabetes.

OBJECTIVES/GOALS: Childhood psychiatric symptoms are highly comorbid. Their co-occurrence and association with negative life outcomes is partially explained by deficits in executive control, or processes enabling self-regulation. Here, we test a novel executive neural target in three fMRI tasks and its relevance to shared psychopathology. METHODS/STUDY POPULATION: We studied 60 children [15 F/45 M; mean age (SD)=11.6 years (1.62)] with diverse diagnoses including attention deficit disorder (n=26) and autism spectrum disorder (n=22). We extracted a latent general factor of psychopathology using principal component analyses applied to parent-report Child Behavior Checklist syndrome scores. Subjects completed 3 executive control fMRI probes, tapping adaptive control, working memory, and inhibition. Correlational psychophysiological interaction (cPPI) analysis measured correlations between executive control-related modulations of activity in 414 network-affiliated parcels. We selected parcels exhibiting control-related cross-network correlations as well as control-related activity across all tasks and tested them for association with psychopathology. RESULTS/ANTICIPATED RESULTS: cPPI connectivity matrices were thresholded and graphs were identified using the Network-Based Statistic toolbox (p90th percentile PC) as well as control-related activation (>10% activated voxels; p DISCUSSION/SIGNIFICANCE: Our results examine cross-network interactions between brain regions during 3 fMRI tasks and their role in explaining individual variation in psychopathology. As executive control links to both comorbidity and life outcomes, identifying the clinically-relevant neural correlates of controlled behavior may lead to transdiagnostic treatments.