bluuebunny/crossref_metadata_2025 huggingface dataset:
309 Measurement of agonist-induced calcium and mechanical signals in human airway smooth muscle cells
Authors: Johnson Santina , Annamdevula Naga , Jalkh Josephine , Ijaz Zara , Britain Andrea L. , Tambe Dhananjay , Francis C. Michael , Deshpande Deepak , Leavesley Silas J. , Rich Thomas C.
Published: April 2023

OBJECTIVES/GOALS: The goal of this proposal is to develop a technology that combines calcium imaging via confocal microscopy, and force measurement via monolayer stress microscopy to perform simultaneous quantitative measurements of agonist-induced Ca2+ and mechanical signals in HASMCs. METHODS/STUDY POPULATION: The methods by which second messenger signals and changes in mechanical forces determine specific physiological responses are complex. Recent studies point to the importance of temporal and spatial encoding in determining signal specificity. Hence, approaches that probe both chemical and mechanical signals are needed. We combine hyperspectral imaging for second messenger signal measurements, monolayer stress microscopy for mechanical force measurements, and S8 analysis software for quantifying localized signals. Imaging was performed using an excitation-scanning hyperspectral microscope. Hyperspectral images were unmixed to identify signals from fluorescent labels and microparticles. Images were analyzed to quantify localized force dynamics through monolayer stress microscopy. RESULTS/ANTICIPATED RESULTS: Results indicate that localized and transient cellular signals can be quantified and mapped within cell populations. Importantly, these results establish a method for simultaneous interrogation of cellular signals and mechanical forces that may play synergistic roles in regulating downstream cellular physiology in confluent monolayers. DISCUSSION/SIGNIFICANCE: We will measure the distribution of chemical and mechanical signals within cells, providing insight into the dynamics of cell signaling. Studies will have implication in the understanding of infections, drug delivery in which non-uniform distributions of drugs are a certainty, and in understanding coordinated responses in cellular systems.
313 Microstructural changes in the brainstem regions of the auditory pathway among children with hearing loss
Authors: Ahmad Iram , Moon Peter , Ward Kristi , Din Taseer , Saki Sara , Cheng Alan , Yeom Kristen
Published: April 2023

OBJECTIVES/GOALS: Diffusion MRI can identify microstructural brain changes and can provide insight into neural development and to potentially prognosticate speech and language outcomes in children with SNHL. The goal of our study was to investigate MRI-based microstructural changes along the brainstem regions of the auditory pathway in pediatric patients with SNHL. METHODS/STUDY POPULATION: We reviewed cohort of pediatric patients with SNHL who obtained MRI at 3T between 2011 and 2019. We identified 16 pediatric patients (age RESULTS/ANTICIPATED RESULTS: We identified significant differences in FA values of the SON between the SNHL cohort and controls (0.377 ± 0.056 vs 0.422 ± 0.052; p=0.009). No other FA or MD values were significantly different between the two groups. Among younger children (ï‚£5 years), MD was significantly decreased in the SNHL cohort compared to controls in the IC (0.918 ± 0.051 vs 1.120 ± 0.142; p5 years), there were no significant differences in MD (1.124 ± 0.198 vs 0.997 ± 0.103; p = 0.119). There were no significant differences in MD or FA in the white matter fibers of the IC-SON tract. DISCUSSION/SIGNIFICANCE: This study is the first to assess microstructural changes in brainstem auditory pathway regions among children with SNHL. Longitudinal studies are warranted to assess the predictive value of these MRI-based findings for long-term outcomes and the efficacy of intervention.
321 Paired associative stimulation: a tool for assessing sensorimotor neural signaling and lower limb function post-stroke
Authors: Cash Jasmine , Heise Kirstin-Friederike , Kindred John , Bowden Mark
Published: April 2023

OBJECTIVES/GOALS: A stroke can impair neural communication between sensory and motor pathways thus compromising walking function. Paired associative stimulation (PAS) is a useful assay of sensorimotor integration (SMI) with limited use post-stroke. The objective of this study will be to determine lower extremity PAS effectiveness and reliability post-stroke. METHODS/STUDY POPULATION: This study will use a pre-post, cross-sectional design. Ten healthy controls and 10 individuals with chronic stroke (>6 months) will be recruited. PAS protocols will be individualized to account for between-subject variability in sensorimotor signaling by first measuring cortical sensory signaling using electroencephalography. Post-stroke participants will then receive PAS targeting the paretic tibialis anterior muscle; healthy controls will receive PAS targeting the non-dominant TA. Changes in cortically derived muscle responses will be characterized by absolute motor-evoked potential amplitude (MEPAmp) change, elicited by transcranial magnetic stimulation, over two sessions separated by >24 hours. Clinical measures of sensorimotor function and walking ability will also be performed. RESULTS/ANTICIPATED RESULTS: By individualizing PAS protocols, we expect to see significant increases in MEPAmp pre to post PAS, determined using paired t-tests. We also anticipate reliable PAS-induced increases in MEPAmp, which will be assessed using two reliability statistics: intraclass correlation coefficient and coefficients of variation of method error. Lastly, the increases in MEPAmp will be correlated with measures of sensorimotor function and walking ability, anticipating that greater increases in MEPAmp will be related to better walking ability and sensorimotor functioning. Correlations will be assessed via a Pearson’s correlation. A preset alpha = 0.05 will be used to determine significant findings. DISCUSSION/SIGNIFICANCE: The importance of this study is that establishing individualized PAS protocols could potentially provide a reliable and clinically relevant measure of SMI. Understanding post-stroke lower extremity SMI is necessary for furthering targeted and personalized interventions to combat walking deficits.
323 Perceived Stress and Access to Care in Parents of Children Living a Diagnostic Odyssey in Puerto Rico
Authors: Albino Elinette M , Martinez Karen , Carlo Simon , Chapel-Crespo Cristel , Ortiz Antonio , Santiago-Cornier Alberto , Velez-Bartolomei Frances , Buxo Carmen
Published: April 2023

OBJECTIVES/GOALS: Diagnostic odyssey is the time it can take to a patient for receiving a diagnosis. Diagnostic process in rare diseases can be complex due to the heterogeneity of symptoms and lack of access to care. We aim to evaluate the association between diagnostic odyssey, perceived stress, and access to care, in parents of Puerto Rican patients with a rare disease. METHODS/STUDY POPULATION: We propose a cross-sectional study in parents of 100 children who received an uninformative whole exome sequencing (WES) report during a scheduled appointment with their geneticist. Discussion of WES results during clinical session, followed by a Perceived Stress Scale (PSS-10) and semi-structured interview to explore the experience of access to care during the diagnostic process will be arranged. Observation and interviews will be recorded. Data analysis and descriptive statistics will be calculated using STATA. Statistical associations (OR) will be estimated using generalized linear models at a 5% significance level. RESULTS/ANTICIPATED RESULTS: We expect to find high perceived stress in parents of Puerto Rican pediatric individuals having rare diseases, especially among single mothers. We will be able to identify limited access to care in Puerto Rico, especially in the testing pre-authorization process and long waits for geneticist appointments. Demand for advanced diagnostics is above the number of medical geneticists available in Puerto Rico, which triggers delayed diagnosis, management, and counseling. Therefore, these could affect the health disparities in our population with rare diseases. DISCUSSION/SIGNIFICANCE: This descriptive study will evaluate perceived stress in parents of pediatric patients living a diagnostic odyssey in Puerto Rico. No study has described perceived stress and access to care in this Hispanic population with undiagnosed conditions. Findings will contribute to a deep understanding of diagnostic process and limited access to care.
330 Remote Monitoring of Pediatric Heart Failure: A Systematic Review
Authors: Dasari Saisamhitha , Gopinath Bhavya , Gaulke Carter , Patel Sunny
Published: April 2023

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.
340 The Effects of PTSD-Dependent Neurogenic Hypertension and Inflammation on Thoracic Aortic Aneurysm Progression
Authors: Holman Heather , Sidles Sara , Xiong Ying , Rinker Jennifer , Ruddy Jean Marie , LaRue Amanda , Mukherjee Rupak , Jones Jeffrey
Published: April 2023

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.
342 The Implications of High Expression of VISTA, a Negative Check Point Regulator, on Prognosis Across Malignant Solid Tumors: a Systematic Review and Meta-Analysis
Authors: Martin Alexander S , Salei Yana V , Lalla Mumtu , Sebastiani Paola , Morin Rebecca A. , Martell Robert E , Trinquart Ludovic
Published: April 2023

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-naÃ¯ve 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 IÂ²coefficient. 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.
343 The Influence of Dynamic Data in Adult Spinal Deformity Surgery Planning and Patient Candidacy: A Preliminary Study
Authors: Fuleihan Antony , Haas Evan , Khalsa Siri , Williams John , Yazdi Youseph , Theodore Nicholas
Published: April 2023

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.
344 The Potential Benefits of Using Senolytics in Colorectal Cancer Treatment
Authors: Gallegos Valerie , Rowdo Florencia Madorsky , White Jessica , Kuo Hui-Hsuan , Podaza Enrique , Martin Laura , Elemento Olivier
Published: April 2023

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.
351 Trends Between Periodontitis and Medial Arterial Calcification in Undiagnosed Type II Diabetes Mellitus
Authors: Dayo Adeyinka , Miles Dale , Boesze-Battaglia Kathleen , Sollecito Thomas , Mupparapu Mel , Corby Patricia
Published: April 2023

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.
355 Unraveling and targeting the innate immune response in Multisystem Inflammatory Syndrome in Children (MIS-C)
Authors: Dick Jenna K , Krishna Venkat , Khaimraj Aaron , Wu Jianming , Orioles Alberto , Cheeran Maxim , Miller Jeffrey S. , Steiner Marie , Hart Geoffrey T.
Published: April 2023

OBJECTIVES/GOALS: The innate immune responses to Multisystem Inflammatory Syndrome in Children (MIS-C) are not fully known. Using samples from MIS-C, we will assess the cellular responses and develop a novel Tri-Specific Killer Engager (TRiKE) that engages innate immune cells to improve those responses. METHODS/STUDY POPULATION: We collected blood samples from 60 pediatric patients from which we isolated plasma and peripheral blood mononuclear cells. We received blood samples from 13 MIS-C, 32 severe acute COVID, 5 COVID-19 asymptomatic, and 15 COVID-19 negative patients. Using plasma, we then performed ELISAs to determine IgG antibody levels against SARS-CoV-2 and plaque reduction neutralization tests to determine neutralizing antibody functions. We isolated DNA to look at Fc receptor genetics. We also utilized utilize flow cytometry assays determine the phagocytosis and killing abilities of the innate cells from these patients. This data will be correlated with clinical outcomes. Additionally, we have developed a novel SARS-CoV-2 TRiKE which directs natural killer (NK) cell killing specifically to of COVID-19 infected cells. RESULTS/ANTICIPATED RESULTS: MIS-C patients had higher IgG antibody titers against SARS-CoV-2 compared to children with symptomatic or asymptomatic COVID. MIS-C patients also neutralized SARS-CoV-2 more effectively than children with acute symptomatic or asymptomatic COVID-19. We found natural killer cells and monocytes are dysfunctional in MIS-C patients and do not kill SARS-CoV-2 infected cells as well. Specifically, NK cells do not kill COVID-19 infected cells as well. To combat this, we have successfully generated and are now testing a Tri-Specific Killer engager (TRiKE) which binds one ends to NK cells, one end to the Spike protein on COVID-19 infected cells and contains IL-15 to improve NK cell function. We anticipate that we can improve NK cell killing of COVID-19 infected cells with this TRiKE. DISCUSSION/SIGNIFICANCE: We found that MIS-C patients have antibodies that can neutralize SARS-CoV-2 but that that innate immune cells that engage antibodies are dysfunctional. We are have successfully developed and are targeting this response with a TRiKE to improve innate immune cell functional; this may serve as an adjunctive therapeutic if proven successful.
356 Upregulated Genes in Age-Related Lobular Involution Stagnation Represent Potential Biomarkers That Link To Increased Breast Cancer Risk
Authors: Radisky Derek , Lue Jaida , Stallings-Mann Melody
Published: April 2023

OBJECTIVES/GOALS: Age-related lobular involution (LI) is a physiological process of breast epithelial regression that occurs primarily during perimenopause (ages 45-55); women in this age range for which the process of LI is delayed, defined as LI stagnation, show significantly increased risk of breast cancer as compared to LI progression patients. METHODS/STUDY POPULATION: The Mayo Clinic Benign Breast Disease (BBD) cohort includes ~1000 women who had multiple sequential benign biopsies. 103 patients were found to have sequential biopsies during the perimenopausal period, of which 10 eventually progressed to breast cancer. These patients were assessed for LI stagnation vs LI progression by quantifying 10 lobules per slide and comparing median acini number and median lobule size between initial and subsequent biopsies from the same patients. RNA was derived from whole tissue sections from the initial biopsies, and profiled using NanoString IO360 and BC360, which were normalized using RUVg methods. Differentially expressed genes associated with LI stagnation were defined as having two-tailed, unpaired p-values less than 0.05. RESULTS/ANTICIPATED RESULTS: Analysis showed subsetting patient sets by time between biopsies improves classification of stagnant vs. progression. Differential gene analysis identified 37 genes associated with LI stagnation and LI progression, and 20 of these genes were found to overlap a set of 128 gnese that were differentially expressed between women who subsequently developed breast cancer vs remained cancer-free. These genes represent potential biomarkers of processes that link LI stagnation and increased breast cancer risk. DISCUSSION/SIGNIFICANCE: In future studies, we intend to study these genes that were shown to be upregulated in LI stagnation for their association with subsequent development of breast cancer in independent cohorts of women with BBD. We will use this knowledge to improve individualized risk assessment, which will help focus surveillance and prevention strategies.
359 Utilization of machine learning approaches on multimodal and ambulatory data to predict individualized symptom course in adults with obsessive-compulsive disorder.
Authors: Frank Adam C , Chang Wellington , Li Ruibei , Narayanan Shrikanth , Peterson Bradley
Published: April 2023

OBJECTIVES/GOALS: This study will collect multimodal and longitudinal data in adults with obsessive-compulsive disorder and healthy controls. A mixed effects random forest machine learning approach will be taken to develop a model that can predict individualized longitudinal OCD symptom burden. METHODS/STUDY POPULATION: Baseline resting state functional MRI (rsfMRI) and measures of symptom burden will be collected in adults with OCD and healthy controls. Longitudinal measures of behavior and physiology–such as heart rate, activity, and sleep metrics - will be collected using Fitbit Charge 5 tracker. Daily assessments of symptom burden and functional status will be collected through a smartphone app. Individuals with OCD will start pharmacotherapy during the study period and all participants will be followed for a total of 10 weeks. Repeat rsfMRI imaging will occur at study conclusion. Data will be analyzed using a mixed effects random forest machine learning algorithm with assessment of model performance. RESULTS/ANTICIPATED RESULTS: Prior studies of symptom severity in psychiatric illness and affect in non-clinical populations have found longitudinal features - such as lexical and acoustic measures, participant context, heart rate, and sleep metrics–that were predictive of these states over time. It is anticipated that the present study will extend these results to individuals with OCD and identify physiologic and behavioral features that track personalized symptom burden longitudinally in this patient population. A model able to predict when symptoms are elevated could allow for provision of additional treatment or interventions targeted to times of high symptom burden. DISCUSSION/SIGNIFICANCE: This study will be the first to collect and analyze longitudinal measures of behavior, symptoms, and physiology in patients with OCD with a goal of predicting symptom burden. Identification of elevated symptom burden would allow for implementation of just-in-time treatment, during these periods.
361 WDR5 represents a therapeutically exploitable target for cancer stem cells in glioblastoma
Authors: Hubert Christopher , Mitchell Kelly , Sprowls Samuel , Shakya Sajina , Arora Sonali , Silver Daniel J. , Goins Christopher M. , Wallace Lisa , Roversi Gustavo , Schafer Rachel , Kay Kristen , Miller Tyler E. , Lauko Adam , Bassett John , Kashyap Anjali , Kass J. D’Amato , Mulkearns-Hubert Erin E. , Johnson Sadie , Alvarado Joseph , Rich Jeremy N. , Paddison Patrick J. , Patel Anoop P. , Stauffer Shaun R. , Hubert Christopher G. , Lathia Justin D.
Published: April 2023

OBJECTIVES/GOALS: Glioblastomas (GBMs) are heterogeneous, treatment-resistant tumors that are driven by populations of cancer stem cells (CSCs). In this study, we perform an epigenetic-focused functional genomics screen in GBM organoids and identify WDR5 as an essential epigenetic regulator in the SOX2-enriched, therapy resistant cancer stem cell niche. METHODS/STUDY POPULATION: Despite their importance for tumor growth, few molecular mechanisms critical for CSC population maintenance have been exploited for therapeutic development. We developed a spatially resolved loss-of-function screen in GBM patient-derived organoids to identify essential epigenetic regulators in the SOX2-enriched, therapy resistant niche. Our niche-specific screens identified WDR5, an H3K4 histone methyltransferase responsible for activating specific gene expression, as indispensable for GBM CSC growth and survival. RESULTS/ANTICIPATED RESULTS: In GBM CSC models, WDR5 inhibitors blocked WRAD complex assembly and reduced H3K4 trimethylation and expression of genes involved in CSC-relevant oncogenic pathways. H3K4me3 peaks lost with WDR5 inhibitor treatment occurred disproportionally on POU transcription factor motifs, required for stem cell maintenance and including the POU5F1(OCT4)::SOX2 motif. We incorporated a SOX2/OCT4 motif driven GFP reporter system into our CSC cell models and found that WDR5 inhibitor treatment resulted in dose-dependent silencing of stem cell reporter activity. Further, WDR5 inhibitor treatment altered the stem cell state, disrupting CSC in vitro growth and self-renewal as well as in vivo tumor growth. DISCUSSION/SIGNIFICANCE: Our results unveiled the role of WDR5 in maintaining the CSC state in GBM and provide a rationale for therapeutic development of WDR5 inhibitors for GBM and other advanced cancers. This conceptual and experimental framework can be applied to many cancers, and can unmask unique microenvironmental biology and rationally designed combination therapies.
363 Association between self-monitoring and ketogenic diet adherence in a technology-assisted lifestyle intervention
Authors: Li Shiyu , Du Yan , Wang Jing , Li Chengdong , Sharma Kumar
Published: April 2023

OBJECTIVES/GOALS: Self-monitoring (SM) improves adherence to low-fat low-calorie (LFLC) diet for weight management. Ketogenic diet (KD) is a promising alternative to LFLC, however, it is unclear whether SM improves KD adherence. We examined the association between SM and KD adherence during the first 12 weeks of a 6-month technology-assisted lifestyle intervention. METHODS/STUDY POPULATION: We included 30 (50.8 ± 12.4 years, 70% female) overweight/obese (body mass index: 37.1 ± 7.2 kg/m2) participants in the analysis. They received personalized KD goals with very low-carbohydrate (22–62 g/d), moderate protein (52 -87 g/d), and high-fat (115 - 219g/d) and calorie intake goals (1338–2554 kcal/d). Additionally, participants performed daily diet, exercise, and weight SM. Adherence to KD was measured by (1) self-monitored dietary intake, and (2) percent of days in ketosis state (blood ketone≥0.5 mmol/L) captured by a fingerstick blood ketone meter. SM frequency was defined as percent of days participant logged food intake, wore fitness tracker, and weighed body weight. Pearson correlation coefficients were computed to examine the correlation between SM in diet, exercise, and weight with KD adherence. RESULTS/ANTICIPATED RESULTS: Percentage of days participants SM for diet, exercise, and weight was 58.4 ± 32.2%, 66.4 ± 30.9%, and 59.0 ± 32.6%, respectively. Correlational analysis more frequent diet SM was positively correlated with more days in ketosis (r = 0.58, p = 0.003), higher fat intake (r = 0.68, p = 0.0001), and higher calorie intake (r = 0.67, p = 0.002) within the fat and calorie goals set; more frequent weight SM was positively correlated with more days in ketosis (r = 0.48, p = 0.02), higher fat intake (r = 0.45, p = 0.023), and higher calorie intake (r = 0.44, p = 0.027). DISCUSSION/SIGNIFICANCE: We found that diet and weight SM were positively associated with fat and calorie intake, as well as days in ketosis. Given the reported promising effect of KD on weight loss and the challenges of adhering to KD, our findings suggested that promoting SM on diet and weight might be a promising avenue for improving KD adherence leading to successful weight loss.
368 Development of Interactome Network to Understand Adverse Drug Reactions for Antidepressant Medications
Authors: Patel Beenish , Wang Yu-Ping
Published: April 2023

OBJECTIVES/GOALS: Patient adherence with antidepressant therapy is a critical aspect of clinical management. Drug molecules often interact with other off-target proteins resulting in adverse events. We utilized an interactome model for predicting the likelihood of adverse drug reactions in order to accurately prescribe antidepressant medications to a patient. METHODS/STUDY POPULATION: We utilized an interactome model to study physical interactions between proteins and biological functions disrupted by major depression to understand how changes in genes alter patient-specific drug efficacy and cause adverse drug-reactions. To study how drugs and diseases propagate through the proteins and biological functions, we harnessed diffusion profiles to represent nodes in the graphical model and used a biased random walks model to illustrate how signals propagate in a heterogeneous biological network. The edge weights were defined for the drug, disease, protein, and biological function node types. The interactions studied included drug-protein, disease-protein, protein-protein, protein-biological function, biological function-biological function. RESULTS/ANTICIPATED RESULTS: Based on previous studies, we anticipate that the genetic variants modulating antidepressant response include 5-HTT, STin2, HTR1A, HTR2A, TPH1 and BDNF. Additionally, genetic variants such as SLC6A4 as well as the HPA pathway may play an important role in antidepressant therapeutic response. With regards to medication drug response, the genes, SLC6A4 and HTR2A, have been known for encoding proteins that affect the synaptic cleft of serotonergic neurons. Also, the gene SLC6A4 has been shown to encode for the serotonin reuptake transporter, which is the main pharmacological target of SSRIs. In female patients with major depression, the polymorphism associated with MAOA gene may be involved in the pharmacological response. DISCUSSION/SIGNIFICANCE: The key to successful depression treatment is early adherence but nearly 60% of patients discontinue antidepressants within three months. By developing a strong understanding of the genes that alter treatment efficacy, we can provide patients with more awareness regarding the most effective treatment plan to minimize adverse events.
369 Differentiating Dementia with Lewy Bodies and Alzheimer’s Disease Using Extracellular Vesicles
Authors: Bregendahl Maria , Faroqi Ayman , Zhang Zhengrong , You Yang , Ikezu Tsuneya , De Souza Suelen Boschen Lucio , McLean Pamela
Published: April 2023

OBJECTIVES/GOALS: Extracellular vesicles (EVs) in biofluids can reflect pathology in the brain (like alpha-synuclein protein linked to Dementia with Lewy Bodies) making them attractive disease biomarkers to distinguish disease. Here, we will investigate their potential as non-invasive biomarkers to differentiate Dementia with Lewy Bodies & Alzheimer's Disease. METHODS/STUDY POPULATION: We will leverage the collection of antemortem plasma from autopsy confirmed Dementia with Lewy Bodies (DLB), Alzheimer's Disease (AD) & healthy controls (HC) through the Mayo Clinic Brain Bank. We will characterize antemortem brain-derived plasma EVs (bPEVs) & post-mortem brain-derived EVs (bEVs) of the same patient using nanoparticle tracking, electron microscopy, & western blotting for EV-associated proteins. We will then measure levels of total & phosphorylated alpha-synuclein (asyn) using AlphaLisa. We will assess EVs' ability to initiate asyn aggregation using a real-time quaking-induced conversion (RT-QuIC) assay & a cell-based Fluorescence Resonance Energy Transfer (FRET) assay. Once characterized, this will allow differentiation of bpEVs in DLB, AD, or healthy controls. RESULTS/ANTICIPATED RESULTS: We hypothesize that asyn levels and seeding capacity will discriminate DLB from AD. With AlphaLisa assay, we expect higher total and phosphorylated asyn levels in DLB bpEVs and their corresponding post-mortem bEVs. Using RT-QuIC and FRET assay, we expect EVs isolated from DLB patient samples to support seeded aggregation, whereas EVs from AD and HC will not. DISCUSSION/SIGNIFICANCE: To date, no diagnostic or less invasive biomarkers can distinguish DLB from AD. The successful completion of the aims outlined in this proposal will identify characteristics of bpEVs that differentiate DLB from AD or HC and support the development of bpEVs as a non-invasive, early biomarker to diagnose patients presenting with dementia from DLB.
373 Identification of potential targets for immunotherapy in a cynomolgus macaque model of Ebola virus disease
Authors: Wanninger Timothy , Saldarriaga Omar A. , Millian Daniel E. , Comer Jason E. , Khanipov Kamil , Golovko George , Paessler Slobodan , Stevenson Heather L.
Published: April 2023

OBJECTIVES/GOALS: Ebola virus infection causes severe disease and liver injury in humans. Macrophages contribute to inflammatory signaling and are prevalent in the liver. We assessed the activation status, including therapeutic target expression, of hepatic macrophages. METHODS/STUDY POPULATION: We compared formalin-fixed, paraffin-embedded liver tissue from terminal Ebola virus-infected and uninfected control cynomolgus macaques, a gold-standard model for human disease. We characterized region-specific protein and whole transcriptome expression in these tissues using GeoMx Digital Spatial Profiling. Macrophage (CD68+) and leukocyte (CD45+) accumulation in liver tissue was quantified by immunofluorescence image analysis using digital pathology software. RESULTS/ANTICIPATED RESULTS: Macrophage-specific (CD68+) regions in the liver of Ebola virus-infected macaques demonstrated a shift towards an inflammatory gene expression profile, as compared to those from healthy control tissue. These regions showed differential expression of monocyte/macrophage differentiation, antigen presentation, and T cell activation gene sets, which were associated with decreased MHC-II allele expression. Moreover, macrophage-specific regions in the infected macaques showed enriched expression of genes or proteins associated with known immunomodulatory therapeutics, including S100A9, IDO1, and CTLA-4. DISCUSSION/SIGNIFICANCE: These data demonstrate that hepatic macrophages express an inflammatory phenotype, that their ability to present antigens to the adaptive immune system may be impaired, and that they express therapeutically targetable markers for immunomodulation of these cells during Ebola virus infection.
374 Identification of Psychosis Risk Biomarkers in 22q11DS for future translational studies
Authors: Parker David , Imes Sid , Ruben Gabrielle , Cuthbert Bruce , Hershey Brett , Walker Elaine , Ousley Opal , Cubells Joseph , Duncan Erica
Published: April 2023

OBJECTIVES/GOALS: 22q.11 deletion syndrome (22q11DS) is a genomic syndrome that elevates risk for psychosis >20-fold. We used a battery of cognitive and psychophysiological psychosis-risk biomarkers in 22q11DS patients and healthy subjects in order to identify biomarkers of psychosis in 22q11DS that could be used as translational targets in intervention studies. METHODS/STUDY POPULATION: We recruited 15 22q11DS individuals (Mean age=30, M/F=9/6) and 19 healthy controls (HCs; Mean age=34, M/F=5/14). Each individual completed the MATRICS Consensus Cognitive Battery (MCCB), the Wechsler Abbreviated Scale of Intelligence, Second edition (WASI-II) Verbal IQ subtests, and the computerized Wisconsin Card Sorting Task (WCST). To examine auditory EEG responses, each participant completed the 'Double-Deviant' target detection paradigm, which presents a pseudorandom sequence of frequent standard tones and infrequent deviant tones. Mismatch negativity (MMN) metrics were generated from this assessment. Welch's t-tests were completed for neurocognitive variables. One-Way ANOVAs were completed to examine EEG results, with sex entered as a separate factor and age entered as a covariate. RESULTS/ANTICIPATED RESULTS: Significant group differences were found in 8 of the 9 neurocognitive measurements (FDR-adjusted p's< 0.02, average Cohen's d=1.62, average observed power= 0.91) indicating widespread cognitive deficits in 22q11DS subjects across multiple domains. The Double-Deviant MMN ERP response was significantly smaller in absolute magnitude in the 22q11DS group (FDR-adjusted p=0.048, Cohen’s d= -0.864, observed power= 0.58). The MMN ERPs for the frequency and duration deviants were not significantly different (FDR-adjusted p's> 0.33). No group by sex interactions were observed in any of the measures. Neurocognitive variables were associated with psychosis positive, negative, general, and disorganized symptom scales. DISCUSSION/SIGNIFICANCE: Our results identify potential psychosis-risk biomarkers in 22q11DS. If replicated, these biomarkers could provide important translational targets for future clinical trials for individuals with 22q11DS and other individuals at-risk for psychosis syndromes.
376 Neurologic Deterioration in Children with Non-Severe Traumatic Intracranial Hemorrhage: A Multicenter Cross-Sectional Study
Authors: Chaudhari Pradip P. , Durham Susan , Bachur Richard G. , Pineda Jose , Khemani Robinder G.
Published: April 2023

OBJECTIVES/GOALS: Children with traumatic intracranial hemorrhage are monitored closely for deterioration and need for intervention. Data on risk factors for deterioration in nonsevere head injury are limited. Our objective was to identify children with hemorrhage from non-severe head injury who are at risk for deterioration. METHODS/STUDY POPULATION: We conducted a 10-site cross-sectional study of children 8. Our primary outcome was clinically important hemorrhage after injury and within 96 hours of ED arrival, defined as ED interventions (intubation, hyperosmotic agents, or neurosurgery within 4 hours of arrival) or clinically important deterioration (new or worsening signs/symptoms with an acute change in management). After testing model assumptions, we used logistic regression to identify clinical and neuroradiographic factors associated with clinically important hemorrhage. RESULTS/ANTICIPATED RESULTS: We studied 763 children with intracranial hemorrhage, with a median (IQR) age of 3.0 (0.4, 10.5) years. Initial GCS was mild (14-15) in 89.4% (n=682) and moderate (9-13) in 10.6% (n=81). Clinically important hemorrhage was observed in 19.5% (n=149), and 7.8% (n=59) developed clinically important deterioration. Median (IQR) time to deterioration was 17.6 (4.6, 37.9) hours. In our sample, 16.3% (n=124) underwent critical interventions, 54.9% (n=419) were admitted to an ICU, and 50.1% (n=382) underwent repeat neuroimaging. We found older age (OR 1.6; 95% CI 1.3, 1.9), lower GCS (OR 5.0; 95% CI 2.9, 8.5), and epidural hemorrhage (OR 3.3; 95% CI 2.0, 5.5) was associated with clinically important hemorrhage. DISCUSSION/SIGNIFICANCE: Clinically important hemorrhage occurred in one in five children with non-severe head injury. Clinical and neuroradiographic factors associated with ED interventions and deterioration were identified. Risk stratification algorithms using these data will be developed to assist clinicians caring for children with head injury.
377 Randomized Placebo-controlled Trial to Test the Efficacy of genetically informed biomarker Nicotine Metabolite Ratio (NMR), and transdermal nicotine replacement therapy (NRT) versus varenicline
Authors: Amadi-Onuoha Obumneke
Published: April 2023

OBJECTIVES/GOALS: The overall aim of the proposed study is to evaluate the effectiveness and clinical utility of the NMR as a biomarker of response to placebo, transdermal nicotine, and varenicline to be utilized within the clinical practice as a point-of-care predictor to tailor an individual’s smoking cessation treatment. METHODS/STUDY POPULATION: A 12-week phase II, stratified multicenter, randomized, placebo-controlled trial of 3 treatment groups to measure treatment-seeking smokers (n-900:150 slow metabolizers; 150 normal metabolizers), randomized to 12-weeks of nicotine patch (active patch + placebo pill), varenicline (active pill + placebo patch), or placebo (placebo pill + patch). At the end of treatment, patients will be followed for 12 months. Study Population: Adult smokers 18–65 years old reported smoking≥10 cigarettes/day for≥6 months (verified by carbon monoxide (CO) >10 ppm). Drug and route of administration: RESULTS/ANTICIPATED RESULTS: The study would conclude the effectiveness of the interventions well defined. DISCUSSION/SIGNIFICANCE: The report will provide robust evidence to support the effectiveness of NRT intervention on smoking cessation.