bluuebunny/crossref_metadata_2025 huggingface dataset:
250 A Living Library for Uveal Melanoma
Authors: Dalvin Lauren A. , Erickson Samantha R. , Pfannkoch Cynthia M. , Miley David R. , Salomao Diva R. , Fautsch Michael P. , Markovic Svetomir N. , Fernandez-Zapico Martin E.
Published: April 2023

OBJECTIVES/GOALS: Overall response rates for metastatic uveal melanoma (UM), regardless of treatment modality, are less than 20%, highlighting an urgent need for novel therapies. Herein, we present a UM patient-derived organoid (PDO) biobank as a novel platform for translational research. METHODS/STUDY POPULATION: Patients with primary choroidal or ciliochoroidal UM undergoing enucleation from 7/1/2019-9/30/2022 were invited to enroll. Tumor tissue was harvested within 30 minutes of globe removal. Cells were isolated using the human tumor isolation kit and gentleMACS dissociation protocol (Miltenyi Biotech). PDOs were placed on Cultrex-coated multiwell plates and cultured in supplemented RPMI media. DNA and RNA were isolated using kits from Zymo Research. Exon-enriched libraries and RNA were sequenced using an Illumina HiSeq 4000. Immunohistochemistry (IHC) assessed the following histone post-translational modifications: H3K4me1/3, H3K27Ac, and H3K27me. RESULTS/ANTICIPATED RESULTS: PDOs were established in 19 of 20 (95%) attempted cases. BAP1 protein expression was retained (n=7) or lost (n=12) in the primary tumors, with matching phenotype confirmed in PDOs. In 9 sequenced cases, a driving mutation was present in GNAQ (n=4), GNA11 (n=4), or CYSLTR2 (n=1). Morphology ranged from spindle-like to epithelioid clusters, mimicking primary tumor histopathology. Pigmentation increased with time in culture. Growth in culture was slow, and 1-2 months were allotted prior to passaging in most cases. Whole exome and RNA-sequencing confirmed distinct molecular profiles, with differential staining of active chromatin marks by IHC. DISCUSSION/SIGNIFICANCE: A biobank of primary UM PDOs with unique morphological and molecular characteristics has been established. These will serve as a model of human disease to facilitate translational research and investigate personalized treatments for patients with UM.
251 A Mixed Methods Study of Patient and Clinician Views and Experiences of Pharmacogenomic Testing for Major Depressive Disorder
Authors: Hendy Katherine , Ellingrod Vicki , Roberts Scott
Published: April 2023

OBJECTIVES/GOALS: Pharmacogenomic testing for major depressive disorder is an expanding area of precision medicine with multiple direct-to-provider tests available. While clinical evidence supporting testing is expanding, there has been little research on the views and experiences of patients and clinicians utilizing this novel intervention. METHODS/STUDY POPULATION: This ongoing study is conducting semi-structured interviews with clinicians and patients exploring their views of the benefits and limitations of pharmacogenomic testing. Qualitative interviews have been conducted with 10 patients and 10 clinicians who have experience with ordering or receiving results within the past 12 months. Interviews are being thematically coded following a modified grounded theory approach using the Dedoose software. Following the principles of exploratory sequential mixed methods design, findings will be used to develop a survey to be administered to prescribing clinicians in both primary care and psychiatry. The survey will examine clinician's knowledge, interest, and concerns about utilizing testing. RESULTS/ANTICIPATED RESULTS: Preliminary analysis of qualitative interviews indicates that both patients and clinicians find that the broader testing process has benefits beyond the test results themselves. Benefits identified by patients include an increased trust in the process of selecting medications, validation of their negative experiences with medications, and improved communication with their provider. Limitations identified by patients include difficulty in accessing test results, and gatekeeping for testing by providers. Benefits identified by clinicians include increased empathy with patients, medication adherence, and improved communication with patients about medication. Limitations identified by clinicians include difficulty with ordering and interpreting test results. DISCUSSION/SIGNIFICANCE: Medication selection is a difficult process for both patients and clinicians. Improvements to clinician-patient communication and medication adherence are important benefits to consider in the adoption of testing. Future research should include these dimensions in assessment of the benefits and limitations of testing.
260 Breast Cancer Biopsy Triage with Ex-Vivo Microscopy for Downstream Analysis
Authors: Behr Madeline , Roberts Brett , Alizadeh Layla , Brown J. Quincy
Published: April 2023

OBJECTIVES/GOALS: In this study, the ability of a pathologist to detect malignancy on digital pseudo-H&E slides obtained via structured illumination microscopy (SIM) imaging of fresh diagnostic breast biopsies was assessed. The speed of imaging and processing was also assessed for potential clinical implementation. METHODS/STUDY POPULATION: This study was conducted in accordance with an Ochsner Medical Center of New Orleans IRB. 200 patients undergoing either stereotactic or ultrasound-guided diagnostic breast biopsies were consented and an additional core from the suspicious lesion was collected for research use. Research biopsies were transported to the lab and stained with DRAQ5 and Eosin-Y and imaged with SIM before being submitted for histology processing. Imaging and digital processing times were recorded. The resulting SIM images and histology slides were given to a pathologist for blind review to assess accuracy. RESULTS/ANTICIPATED RESULTS: The ex-vivo structured illumination microscopy images and subsequent histology slides from 79 research cores have been assessed to date. Some samples were excluded from the total data set and not included in the final assessment due to technical failures of the imaging protocol. Of the current set, the pathologist has a specificity of 88% and a sensitivity of 65%, as well as an NPV of 88% and a PPV of 65%. Staining time for each biopsy was completed within 3 and a half minutes and imaging at 20x magnification took between 4 and 12 minutes, depending on size and implementation of autofocus to the imaging system. Image processing took approximately 5 minutes per biopsy and is a direct function of biopsy size. DISCUSSION/SIGNIFICANCE: Decreased time between cancer suspicion and treatment will improve the prognosis of breast cancer patients. SIM imaging of fresh breast biopsies could ultimately allow primary and secondary histology to be performed simultaneously and minimize histopathology time, thus allowing clinicians and patients to implement treatment course more quickly.
261 Characterization of Glycosylation Patterns of Single IgA Molecules Using Single-Molecule Fluorescence Microscopy
Authors: Rubin Joseph , Stojanovic Milan , Gharavi Ali , Hess Henry
Published: April 2023

OBJECTIVES/GOALS: IgA1 nephropathy, which can lead to kidney failure, is caused by complexes formed between aberrant galactose-deficient IgA1 and antibodies directed against it. Our goals are to characterize shifting glycosylation patterns at the level of single IgA1 molecules and to apply this to patient samples for early detection and understanding of the disease. METHODS/STUDY POPULATION: To characterize glycosylation patterns on single IgA1 molecules, labelled IgA1 in low concentration was physisorbed to borosilicate glass in a fluidic cell and labelled Jacalin was flowed in to bind with Glycans on IgA1. The samples were observed with a Nikon TiE epi-fluorescence microscope. FRET images were created by exciting the Jacalin dye with a blue laser and recording the red emission of the IgA1 dye with an EMCCD camera. FRET emission intensities of individual IgA1 molecules over time were analyzed to determine how frequent and how long Jacalin binds to each of them. The rate of binding is roughly inversely proportional to the amount of abnormal glycans on a given IgA1 molecule. After the method is perfected, we intend to compare the glycosylation patterns of healthy and diseased patient samples. RESULTS/ANTICIPATED RESULTS: Addition of the competitive binder Galactose to the solution led to an increase in the off times of Jacalin and IgA1 and a decrease in the on times in a concentration-dependent manner, yielding an increase in the dissociation constant. Dissociation constants of individual molecules within a single experiment vary by 3 orders of magnitude, which cannot be attributed to stochastic fluctuations but rather reflects differences in the adsorption geometry. Nevertheless, the unaffected dissociation constant can be identified. We expect that when this method is applied to samples from healthy and IgA1 Nephropathy patients, specific IgA1 molecules from patients will have higher dissociation constants for Jacalin compared to those from healthy patients. DISCUSSION/SIGNIFICANCE: The binding rates of Jacalin to single IgA1 vary by 3 orders of magnitude. The observed heterogeneity shows the Jacalin probe can differentiate between different IgA1 populations. An understanding of which IgA1 molecules in patient samples are problematic and what their distribution of Glycans is can lead to discovering biomarkers and treatments.
276 Effect of Practice Schedule on Motor Learning of Protective Stepping Responses in Persons with Parkinson’s Disease and Age-matched Controls.
Authors: Akinlosotu Ruth Y , Westlake Kelly
Published: April 2023

OBJECTIVES/GOALS: Persons with Parkinson’s disease (PwPD) have set-shifting impairments that may limit motor learning of protective responses to prevent falls. This study compares the effect of blocked versus random practice schedules on acquisition and retention of protective stepping stability in PwPD & age-matched controls. METHODS/STUDY POPULATION: Twenty PwPD & 20 age-matched controls will be randomly assigned to random or blocked practice groups to experience slip and trip-like perturbations which induce stepping responses. Blocked practice is repeating a task (e.g., slip) several times before the next task, while random practice is randomly practicing two or more tasks (e.g., slips & trips). Each subject will wear a safety harness to prevent falls, each fitted with a load cell that indicates when a fall would have occurred. We will assess the anterior-posterior and mediolateral margin of stability at 1st protective step, step length, and in-task fall incidence before, 10 minutes, and two days after practice. We will also compare outcomes between participants in the blocked vs. random practice schedule using two ANOVA. RESULTS/ANTICIPATED RESULTS: To date, we have recruited three control subjects. We anticipate that blocked practice will be superior to random practice for protective stepping acquisition and retention PwPD. Existing literature on motor learning of voluntary tasks indicates that PwPD have difficulty acquiring and retaining motor skills via random practice due to the higher cognitive processing required than blocked practice. Furthermore, since the basal ganglia’s set-shifting role is crucial for random practice, it is expected that random practice will limit protective steps acquisition and retention in PwPD. In healthy control, while blocked practice is better for acquisition, random practice will be superior for retention. DISCUSSION/SIGNIFICANCE: The study’s findings will provide knowledge that may foster the development of robust balance rehabilitation protocols to improve postural responses in PwPD and reduce delayed and ineffective stepping responses that results in falls among PwPD.
279 Electronic Health Record Data and Topological Data Analysis to Predict Clinical Outcomes Post Myocardial Infarction
Authors: Awolope Anna , Datta Esha , Ballal Aditya , Izu Leighton T , López Javier E.
Published: April 2023

OBJECTIVES/GOALS: The aim of this study is to analyze electronic health record (EHR) data using Mapper PLUS (MP), a new mathematical model, to classify acute myocardial infarction (MI) patients by risk of major adverse events (AE). We tested MP’s ability to define patient subgroups with distinctive risk for death, heart failure or recurrent MI after revascularization. METHODS/STUDY POPULATION: An EHR retrospective analysis of 797 MI patients and 29 variables (i.e., laboratory tests, imaging, vitals, and clinical traits) collected at the time of hospitalization was conducted. All patients received percutaneous coronary intervention and standard pharmacotherapy. MP analysis produced a multi-dimensional nodal graph of the patients based on similarities found within variables. Two algorithms, Walk Likelihood and Walk Likelihood Community Finder were applied to the graph which formed joint clusters according to spatial distance within nodes. The final output was three clusters for risk level evaluation. Risk level (low vs. high) was relative to the average risk of AEs for the entire cohort one year post MI. RESULTS/ANTICIPATED RESULTS: Of three patient subgroups, one (n= 318) had a >1 fold change for the probability of survival without AE when compared to the overall cohort and thus was defined as the low-risk group. The second group (n=304) had DISCUSSION/SIGNIFICANCE: MP stratifies patients into three groups according to predictive variables which relate to the risk for AE following an acute MI treatment. This is a new topological method for patient classification based on minimal input strictly from pre-collected EHR data. More cohort studies are needed to validate MP to classify patients for precision medicine.
281 Fc gamma receptor crosslinking promotes antibody-induced LILRB4 internalization and immune regulation of acute monocytic leukemia
Authors: Morse Joshua , Gui Xun , Deng Mi , Zhang Cheng Cheng , Zhang Ningyan , An Zhiqiang
Published: April 2023

OBJECTIVES/GOALS: The aims of this study were to: 1) determine the involvement of each Fc gamma receptor isoform in antibody-mediated crosslinking and internalization of LILRB4 in acute monocytic leukemia and 2) elucidate the role of this crosslinking and internalization in antibody-mediated immune regulation of these malignant cells. METHODS/STUDY POPULATION: To determine the involvement of Fc gamma receptors in antibody-mediated complex internalization, we generated acute monocytic leukemia cell lines with CRISPR-Cas9 knockout of each Fc gamma receptor isoform. We tested the effects of each knockout on anti-LILRB4 antibody-mediated internalization by flow cytometry and confirmed our findings with confocal microscopy. To elucidate the role of this crosslinking and internalization in immune regulation of acute monocytic leukemia, we conducted preliminary ELISA-based studies of the inflammatory signaling and cytokine release profiles of wild-type and Fc gamma receptor knockout cells treated with the LILRB4-targeting antibody. RESULTS/ANTICIPATED RESULTS: We have concluded that Fc gamma receptor I (CD64) plays a role in LILRB4 crosslinking and internalization by our anti-LILRB4 antibody and there are also contributions from Fc gamma receptor IIA (CD32A) observed in the absence of CD64 on the cell surface. Preliminary signaling studies have demonstrated that Fc gamma receptor-mediated antibody crosslinking and internalization of LILRB4 decreases anti-inflammatory signaling downstream of LILRB4 as well as pro-inflammatory signaling downstream of Fc gamma receptors, particularly in the absence of CD64 on the cell surface. The immunomodulatory effect of antibody-mediated LILRB4 crosslinking and internalization is being confirmed in follow-up signaling, cytokine release and lymphocyte activation studies. DISCUSSION/SIGNIFICANCE: This study will improve the efficacy of LILRB4-targeting antibody therapeutics for patients suffering from acute monocytic leukemia and help characterize CD64 and CD32A as potential clinical biomarkers for patients undergoing LILRB4-targeting antibody immunotherapeutic treatment, currently in first-in-human clinical trials.
282 Food Intake REstriction for Health OUtcome Support and Education (FIREHOUSE): a Proof of Concept Randomized Clinical Trial
Authors: Kwon Sophia , Lam Rachel , Crowley George , Zeig-Owens Rachel , Schwartz Theresa , Colbeth Hilary , Lou Pompeii Mary , St-Jules David E. , Prezant David J. , Sevick Mary Ann , Liu Mengling , Nolan Anna
Published: April 2023

OBJECTIVES/GOALS: Particulate matter (PM) and metabolic syndrome (MetSyn) increase risk of World Trade Center-Lung Injury (WTC-LI). Mediterranean-type diets have also been found to improve lung function. Fire Department of New York 1st-responders with a high PM exposure at WTC and MetSyn may have improved lung function after a Mediterranean dietary intervention. METHODS/STUDY POPULATION: Food Intake REstriction for Health OUtcome Support and Education (FIREHOUSE) randomized clinical trial (RCT) assessed our hypothesis that a low-calorie Mediterranean (LoCalMed) intervention targeting clinically relevant disease modifiers will improve metabolic risk, subclinical indicators of cardiopulmonary disease, quality of life, and lung function in firefighters with WTC-LI. Primary-outcome targeted a LoCalMed loss of BMI(≥1kg/m2). Secondary-outcomes included lung function, quality of life, and cardiovascular health. Male firefighters with WTC-LI and a BMI>27kg/m2 were randomized to: 1. LoCalMed (n=46); or 2. Usual Care (UC; n=43). Clinicaltrials.gov:NCT03581006. RESULTS/ANTICIPATED RESULTS: LoCalMed’s estimated efficacy on BMI reduction crossed the pre-specified significance boundary on interim analysis compared to UC. In addition, improvements were observed in secondary-outcomes of lung health (FEV1 and FVC), inflammation (WBC), vascular disease (DBP), quality of life (SF-36, health perception) and dietary habits (less cholesterol, carbohydrates, fats, and sweets and increased protein) in the LoCalMed arm. DISCUSSION/SIGNIFICANCE: LoCalMed significantly decreased BMI and alleviated adverse health outcomes in our WTC-exposed first responders. A fully powered RCT is required to determine if this approach is efficacious for the treatment of WTC-associated pulmonary disease, as well as LoCalMed’s generalizability to PM associated disease.
284 Generalizable Machine Learning Methods for Subtyping Individuals on National Health Databases: Case Studies Using Data from HRS, N3C, and All of Us
Authors: Bhavnani Suresh K. , Zhang Weibin , Bao Daniel , Hatch Sandra , Reistetter Timothy , Downer Brian
Published: April 2023

OBJECTIVES/GOALS: While disease subtypes are critical for precision medicine, most projects use unipartite clustering methods such as k-means which are not fully automated, do not provide statistical significance, and are difficult to interpret. These gaps were addressed through bipartite networks and tested for generalizability on three national databases. METHODS/STUDY POPULATION: Data. All participants with self-reported stroke from the 2010 Health and Retirement Study (HRS), with cases (n=798) having one or more 8 depressive symptoms measured by the Centers for the Epidemiological Study–Depression 8 scale, and controls (n=389) with none of those symptoms. The replication data set consisted of independent identically-defined participants (cases=725, controls=190) from 1998 HRS. Method. (1) Bipartite network analysis and modularity maximization to automatically identify patient-symptom biclusters with significance. (2) Rand Index to measure the replicability of symptom co-occurrences in the replication data. (3) ExplodeLayout to visualize and interpret the subtypes. (4) R libraries to generalize the methods, upload them to CRAN, and then tested on the N3C and All of Us platforms. RESULTS/ANTICIPATED RESULTS: The analysis identified 4 depressive symptom subtypes (https://postimg.cc/Ny8YwXJW) which had significant modularity (Q=0.26, z=3.03, P DISCUSSION/SIGNIFICANCE: We developed generalizable methods to automatically identify biclusters, measure the clustering significance, and visualize the results for interpretation. These methods were successfully tested on three national level data bases. Such generalizable methods should accelerate the analysis of subtypes, and the design of targeted interventions.
289 Heart Rate Variability in Opioid Use Disordered Participants Undergoing Buprenorphine-Assisted Detoxification
Authors: Russell Lauren , Weaver Jackson , Mancino Michael , Addicott Merideth , Larson-Prior Linda , Oliveto Alison
Published: April 2023

OBJECTIVES/GOALS: This study explored whether gabapentin (GBN) differentially impacted heart rate variability (HRV) and whether HRV was associated with opioid withdrawal ratings among participants with opioid use disorder (OUD) undergoing a randomized, double blind placebo-controlled, trial (RCT) of GBN during a buprenorphine (BUP)-assisted taper. METHODS/STUDY POPULATION: Participants (ages 18-64) with OUD, no recent use of benzodiazepines/barbiturates, and no major psychiatric disorders or unstable medical conditions were enrolled in the RCT, inducted onto BUP starting week 1 day 1 and randomly assigned to receive adjunct GBN or placebo starting week 1 day 3. All participants began a 10-day BUP-taper beginning week 2 day 3. HRV measures were assessed on week 1 day 2 (before GBN/placebo induction), week 2 day 2, and week 3 day 5 (end of BUP taper). HRV metrics were analyzed using Two Sample T-Test to determine differences between GBN vs. Placebo. Correlations between HRV metrics and opioid withdrawal ratings administered at the above timepoints will be analyzed using Spearman correlation. RESULTS/ANTICIPATED RESULTS: 28 participants underwent at least 1 HRV session that resulted in usable data. Preliminary statistical analyses revealed that HRV trended lower for GBN subjects during PB exercises than Placebo subjects, demonstrated by a higher mean heart rate for GBN subjects compared to Placebo subjects (p=0.0506) at the end of the BUP-taper (week 3 day 5). We expect future analyses to demonstrate a negative correlation between certain HRV metrics indicative of parasympathetic tone and opioid withdrawal rating assessment scores indicative of withdrawal severity. Such findings would demonstrate an association between opioid withdrawal severity and lower parasympathetic tone and HRV. DISCUSSION/SIGNIFICANCE: Individuals with OUD have previously been shown to have a lower parasympathetic tone than individuals without OUD. Additionally, opioid withdrawal has been shown to be associated with reduced parasympathetic tone. Our initial findings suggest that adjunct GBN administration was not associated with lower parasympathetic tone during PB exercises.
291 High Potency STING Agonists Induce Adaptive Immunity-Dependent Curative Responses in an Immune Checkpoint Blockade-Refractory Glioblastoma Model
Authors: Lea Spencer , Chen Chao-Hsien , Wei Jun , William Ivana , Curran Michael
Published: April 2023

OBJECTIVES/GOALS: Glioblastoma (GBM) is the most common and aggressive adult primary brain malignancy. Clinically, GBM is refractory to T cell immune checkpoint blockade (ICB), in part due to its dense immune suppressive myeloid stroma. Here we show that myeloid-targeting STING agonists can repolarize the GBM microenvironment to cure ICB-refractory GBM models. METHODS/STUDY POPULATION: Using the synthetic cyclic di-nucleotide STING agonist IACS-8803 (8803) we treated orthotopic ICB-refractory QPP8 orthotopic murine GBM tumors intratumorally. We then analyzed survival and performed high parameter flow cytometry profiling of the tumor immune microenvironment following STING agonist treatment. To assess the contribution of adaptive immunity to STING agonist therapeutic efficacy, we treated orthotopic QPP8 tumors implanted in RAG1 KO mice and monitored survival. RESULTS/ANTICIPATED RESULTS: We found that STING agonist therapy cured murine orthotopic QPP8 tumors, in contrast to ICB that showed no survival benefit. In RAG1-/- mice bearing QPP8 tumors STING agonist therapy extended survival, however, the curative effect observed in wild-type mice was lost in the absence of adaptive immunity. STING agonist-treated QPP8 tumors displayed increased counts of CD8 T cells and NK cells, and decreased CD8 T cell PD1 expression. Infiltration of STING-treated gliomas by Ly6C+ F4/80+ Mono-MDSC substantially increased; however, these cells expressed reduced CD206 and CD163, suggestive of reduced immuno-suppression. Finally, in the cervical LN of QPP8-treated mice the frequency and CD80/CD86 expression of cDC1 cells increased.â€‹ DISCUSSION/SIGNIFICANCE: ICB has failed in GBM, and the suppressive myeloid stroma remains a major barrier to generating anti-GBM T cell responses. Our work shows that STING activation, which primarily targets innate immunity myeloid cells 'upstream' of T cells in the antitumor immunity cycle, can cure ICB-refractory GBM tumors in an adaptive immunity-dependent manner.
292 Hyperorality in Frontotemporal Dementia: Psychiatric and Neural Correlates Across the Disease Course
Authors: Morrow Christopher , Kamath Vidyulata , Onyike Chiadi
Published: April 2023

OBJECTIVES/GOALS: To describe cognitive and psychiatric symptom profiles of individuals with bvFTD and hyperorality. We test two hypotheses: (1) individuals with hyperorality show more severe psychiatric profiles and (2) neuroanatomic correlates of hyperorality in advanced bvFTD differ from those with early bvFTD. METHODS/STUDY POPULATION: Participants were enrolled in ALLFTD–a multi-site longitudinal study in FTD. We selected the 354 participants who had a primary clinical diagnosis of bvFTD, 344 of whom had data on hyperorality. Each participant underwent extensive clinical interviews and examinations, structural neuroimaging, and blood sampling. Five anatomic regions of interest were identified and analyzed based on previously identified neuroanatomic correlates of hyperorality. Differences in participant characteristics and clinical outcomes were compared using t-tests for continuous variables and PearsonÏ‡2 tests for categorical variables. Linear multivariate regression controlling for age and total intracranial volume (TIV) was used to examine associations between atrophy in regions of interest and hyperorality status. RESULTS/ANTICIPATED RESULTS: Early-stage participants with hyperorality had poorer self-monitoring, empathic concern, and perspective taking as well as higher CDR behavioral subscale scores compared to those without hyperorality. Advanced stage participants with hyperorality had higher scores on the Social Behavior Observer Checklist compared to those without hyperorality. Early-stage participants with hyperorality displayed higher rates of ritualistic/compulsive behavior and motor disturbance. Advanced stage participants had higher rates of apathy, ritualistic/compulsive behavior, anxiety, and elation. In the advanced stage participants, hyperorality was associated with atrophy in the right dorsal striatum, the right ventral striatum, and the right insula cortex. DISCUSSION/SIGNIFICANCE: Hyperorality emerges early and is accompanied by neuropsychiatric symptoms prior to significant neurodegeneration. Overtime, participants with hyperorality develop more psychiatric symptoms as well as atrophy in striatal and insular brain regions. Our findings suggest a role for novel interventions like non-invasive brain stimulation.
296 Identifying Gaps in Elderly Fecal Incontinence Management
Authors: Barlean Braden , Merali Khalil , Smith Meagan , Yang Poplar
Published: April 2023

OBJECTIVES/GOALS: Fecal incontinence is associated with increased caregiver strain, reduced patient dignity and diminished quality of life. A spectrum of incontinence exists, along with a paucity of available solutions for patients and their caregivers. This research aims to stratify this space and identify gaps within the existing solution landscape. METHODS/STUDY POPULATION: To understand this problem, a literature review was performed with key search terms specific to fecal incontinence. These included, anal incontinence epidemiology, fecal incontinence in nursing homes, and incontinence management. To determine gaps within the existing solution landscape, key search terms related to existing solutions for fecal incontinence were also included. These included, fecal management systems, rectal incontinence therapies, and anorectal incontinence procedures.' To perform a population segmentation, white papers, review articles, and cross-sectional studies were reviewed to break down the burden of incontinence in older adults living in nursing facilities and in the community. RESULTS/ANTICIPATED RESULTS: Two unaddressed populations were identified, the first being independent adults over the age of forty, particularly women, who suffer from frequent, bothersome incontinence. These 1.2 million patients are active, living at home, and they restrict their daily activities due to incontinence. However, there are several durable and effective solutions for patients who have sufficient sphincter tone or who are surgical candidates. The second population identified are caregiver dependent older adults residing in nursing facilities who suffer from severe incontinence. This population of 160,000 is affected more severely by consequences of fecal and are poorly served by solutions that are largely absorptive such as diapers and pads. DISCUSSION/SIGNIFICANCE: Although two populations were identified, caregiver dependent older adults residing in nursing homes were identified to have a significant unmet need within incontinence care. Current solutions are onerous and transient, preventing ease and duration for use by caregivers and nurses.
297 Identifying Opportunities and Challenges for Translational Informatics Approaches to Real-World Data: A Diabetes Case Study
Authors: Mistry Sejal , Gouripeddi Ramkiran , Facelli Julio C.
Published: April 2023

OBJECTIVES/GOALS: Diabetes is a group of chronic metabolic diseases and significant gaps remain in our understanding of disease etiology, treatment regimens, and diabetes-related complications. The objective of study is to demonstrate how informatics techniques can leverage real-world data for diabetes research and identify barriers for implementation. METHODS/STUDY POPULATION: We evaluated informatics applications of real-world data in diabetes research conducted by the Facelli Research Group. The types of real-world data were categorized into clinical records, diabetes-related repositories, wearable sensors, and other data sources. Translational informatics applications were characterized into thematic groups of 1.) use of electronic health records, registries, and claims and other data sources to generate real-world evidence, 2.) evolution of novel methods to accelerate generation and use of real-world data, and 3.) infrastructure to support the generation and use of real-world data in translational science. A literature review is being conducted to identify additional articles meeting these themes focused on diabetes research. RESULTS/ANTICIPATED RESULTS: 6 research projects were included for analysis. The diabetes-focus spanned type 1 diabetes, type 2 diabetes, and general diabetes mellitus. Informatics methods included machine learning and data mining while real-world data sources included electronic medical records, the Environmental Determinants of Diabetes in the Young (TEDDY) study, continuous glucose monitors, and the U.S. Environmental Protection Agency (EPA) air pollution monitors. Overall, computability of real-world data, linkage of medical concepts to standardized terminologies, volume of data, and adoption of novel artificial intelligence methods were major determinants of successful implementation. Future work will systematically evaluate informatics applications of real-world data in diabetes from the academic community at large. DISCUSSION/SIGNIFICANCE: Translational informatics approaches are poised to leverage real-world data and better understand diabetes etiology, treatment regimens, and diabetes-related complications. By understanding barriers and opportunities for informatics methods, we can expedite translational applications in diabetes research.
298 Identifying vaginal microbiome profiles that influence tenofovir distribution in the female genital tract
Authors: Lantz Alyssa M. , Staley Christopher , Nicol Melanie R.
Published: April 2023

OBJECTIVES/GOALS: An ex-vivo tissue model has been developed to predict target concentrations of tenofovir diphosphate (TFVdp; active metabolite of tenofovir) but has not been utilized to see how vaginal dysbiosis affects TFVdp/dATP exposure in female genital tract (FGT). My central hypothesis is that presence of specific anaerobic bacteria will increase dATP in FGT. METHODS/STUDY POPULATION: De-identified HIV-negative cervical tissues from women undergoing gynecological surgeries will be procured and a punch biopsy will be used to create explants. TFVdp/dATP concentrations were both tested in both aerobic and anaerobic conditions after a 24-hour incubation in tenofovir (TFV) to determine any changes between conditions. TFVdp/dATP in cervical tissue was be measured using LC-MS. Next, media and explants were collected at baseline to characterize donor microbiome for 6 donors. 16S microbiome sequencing was performed on extracted DNA to obtain the relative abundances of each bacteria species present. To test changes in dATP/TFVdp due to the microbiome, explants will be incubated in TFV for 24 hours with Prevotella and Dialister to specifically see how microbiomes dominated by these taxa affect dATP. RESULTS/ANTICIPATED RESULTS: There was no significant difference in TFVdp formation between aerobic and anaerobic conditions after a 24-hour tenofovir incubation (p = 0.2) for 8 donors. dATP was not quantifiable at 24 hours in explants, so explants are being collected before 24hrs during a TFV incubation to determine how quickly dATP depletes after collection. We were able to characterize the donor microbiome in media and tissue at baseline and 24hrs which had inter variability. We did not see any presence of Prevotella or Dialister in any donors. We are working on characterizing bacteria growth over time to see how the donor microbiome would change during a 24 hour experiment in anaerobic conditions. Finally, we anticipate seeing increases in dATP with a Prevotella or Dialister supplemented donor microbiome compared to baseline donor microbiome. DISCUSSION/SIGNIFICANCE: The addition of vaginal dysbiosis to tissue model will increase accuracy of prediction of 100% protective TFVdp concentrations and is likely to provide a translational model that can be used to improve TFV-based PrEP in women and streamline development of future PrEP candidates, bringing more prevention options to women and ending the HIV epidemic.
300 Impact of the type of mechanical circulatory support (MCS) prior to transplant on development of post-orthotopic heart transplantation (OHT) infections
Authors: Trottier Caitlin A. , Strand Andrew , Snydman David , Chow Jennifer
Published: April 2023

OBJECTIVES/GOALS: In 2018, the United Network for Organ Sharing began prioritizing patients on temporary MCS over those on durable MCS for OHT in an effort to prioritize sicker patients and decrease waitlist mortality. We explored the impact of this change by examining if the type of MCS prior to transplant affects the risk of post-transplant infection. METHODS/STUDY POPULATION: We will conduct a retrospective cohort study of approximately 350 patients that have undergone OHT at Tufts Medical Center between January 2014 and July 2021 who survived at least 72 hours post-transplant and have minimum post-transplant follow-up of one year or time to death if before one year. Chart review will determine the type of MCS in place prior to transplant and the occurrence of infections within one year of transplant. Data will also be collected on patient’s age, sex, medical comorbidities, lab values, and open chest management practices. We will examine differences in the incidence rates of a composite outcome (blood stream infection, invasive fungal infection, skin and soft tissue infection of device sites, and mediastinitis) between patients that were on temporary versus durable MCS. RESULTS/ANTICIPATED RESULTS: We anticipate that this study will show a greater frequency of infections of all types in patients that received temporary as compared with durable mechanical circulatory support prior to transplantation. We will use Cox proportional hazards survival models to model multivariable relationships for predictors of infection. DISCUSSION/SIGNIFICANCE: This study will provide insights into the magnitude and type of infectious complications that patients experience after OHT and the impact that type of MCS and other factors have on their outcomes. The data obtained may have implications for choice of mechanical device prior to undergoing OHT surgery as well as antimicrobial prophylaxis.
301 Inhibition of lysine-specific histone demethylase 1A (KDM1A/LSD1) attenuates DNA double strand break repair and enhances efficacy of temozolomide in glioblastoma
Authors: Alejo Salvador , Palacios Bridgitte , Venkata Prabhakar Pitta , He Yi , Li Wenjing , Johnson Jessica , Jayamohan Sridharan , Vadlamudi Ratna , Sareddy Gangadhara
Published: April 2023

OBJECTIVES/GOALS: Glioblastoma (GBM) patients face a poor prognosis. Glioma stem cells (GSCs), a chemo resistant GBM subpopulation, possess enhanced DNA repair and elevated levels of epigenetic modifier KDM1A. This study aims to establish the significance of KDM1A in DNA repair and determine the potential of novel KDM1A inhibitor NCD38 to enhance TMZ efficacy in GSCs. METHODS/STUDY POPULATION: Patient derived GSCs were obtained via IRB-approved protocol from patient samples at UT Health San Antonio. KDM1A knockdown and knockout cells were generated by transduction of validated KDM1A-specific shRNA or gRNA, respectively. Brain bioavailability of KDM1A inhibitor NCD38 was established using LS-MS/MS. Effect of combination of KDM1A knockdown, knockout, or inhibition with TMZ was studied using cell viability, neurosphere, and self-renewal assays. Mechanistic studies were conducted using CUT&Tag-seq, RNA-seq, immunofluorescence, comet, Western blotting, RT-qPCR, homologous recombination (HR) or non-homologous end-joining (NHEJ) DNA repair reporter assays. In vivo efficacy of KDM1A knockdown or inhibitor alongside TMZ treatment was determined using orthotopic murine GBM models. RESULTS/ANTICIPATED RESULTS: KDM1A knockdown, knockout, or inhibition increased efficacy of TMZ in reducing cell viability and self-renewal of GSCs. Pharmacokinetic studies demonstrated KDM1A inhibitor NCD38 is readily brain penetrable. CUT&Tag-seq studies revealed KDM1A is enriched at DNA repair gene promoters. RNA-seq studies suggest KDM1A inhibition reduces DNA double strand break repair gene expression, with these findings validated using RT-qPCR and Western blotting. Knockdown, knockout, or inhibition of KDM1A attenuated HR and NHEJ-mediated DNA repair capacity. Immunofluorescence and comet assay support findings of increased DNA damage in NCD38/TMZ combination treated GSCs. Importantly, KDM1A knockdown or inhibition enhanced efficacy of TMZ and significantly improved survival of orthotopic GBM tumor-bearing mice. DISCUSSION/SIGNIFICANCE: Our results show compelling evidence that KDM1A is essential for DNA repair in GSCs and that KDM1A inhibition sensitizes GBM to TMZ via attenuation of DNA repair pathways. These findings suggest combination of KDM1A inhibitor NCD38 with TMZ could serve as a promising novel therapeutic strategy that can be translated to improve GBM patient outcomes.
305 Leveraging multi-timepoint blood samples to characterize cancer-associated mutations in the blood over time
Authors: Mack Taralynn , Von Beck Kelly , Silver Alexander , Savona Michael , Vanderbilt Alexander Bick
Published: April 2023

OBJECTIVES/GOALS: Clonal hematopoiesis of indeterminate potential (CHIP) is a common age-related condition that confers an increased risk of blood cancer, cardiovascular disease, and overall mortality. Larger proportions of blood cells with the CHIP mutation (clones) lead to worse outcomes. The goal of this study was to characterize CHIP clonal behavior over time. METHODS/STUDY POPULATION: While DNA biobanks have the ability to identify large cohorts of individuals with CHIP, they typically only contain blood from a single timepoint, limiting the ability to infer how CHIP clones change over time. In this preliminary study, we utilized multi-timepoint blood samples from 101 individuals with CHIP in Vanderbilt’s biobank (BioVU) to characterize clonal behavior over time. Using a CHIP gene-specific sequencing pipeline, we were able to characterize each individual’s CHIP mutation(s) and how the fraction of cells with the CHIP mutation expanded/reduced over time. By Spring 2023, we will also include ~300 additional individuals with CHIP in this study. RESULTS/ANTICIPATED RESULTS: CHIP mutations occurred 48% of the time in DNMT3A and 23% of the time in TET2, consistent with previous studies. 21% of individuals had more than one CHIP mutation. The mean difference in time between the two timepoints was 5.2 years (SD=2.9). Surprisingly, we observed both clonal expansion and clonal reduction across timepoints with 30% of DNMT3A, 0.6% of TET2, and 46% of JAK2 clones shrinking over time. The fastest average expansion was seen in TET2 clones (2% growth/year) and the slowest in DNMT3A clones (0.4% growth/year), but there was a significant amount of variation between individuals. In DNMT3A clones, there were no differences observed between loss of function mutations, missense mutations or DNMT3A R882 hotspot mutations. Clonal competition was observed in individuals with multiple driver mutations. DISCUSSION/SIGNIFICANCE: We used multi-timepoint blood samples to quantify the change in CHIP cell fraction over time on a per individual basis and observed novel clonal behavior and competition. Understanding the factors that influence the rate of CHIP progression can lead to personalized disease risk assessment for individuals with CHIP.
311 Mental Illness and the Development of Postoperative Atrial Fibrillation in Transcatheter Aortic Valve Replacement Patients: Trends over Time
Authors: Kolba Natalie , Morrone Jennifer , Dokko Julia , Novotny Samantha , Yang Jie , Tummala Vineet , Agha Sohaib , Yaligar Ashutosh , Parikh Puja B. , Pryor Aurora D. , Tannous Henry J. , Shroyer A. Laurie , Bilfinger Thomas
Published: April 2023

OBJECTIVES/GOALS: The purpose of this retrospective cohort study was to evaluate the impact of mental illness on first-time transcatheter aortic valve replacement (TAVR) and repeat TAVR (viv-AVR) outcomes including postoperative atrial fibrillation (POAF/AFL), as well as trends over time. METHODS/STUDY POPULATION: Using de-identified data reports from the New York Statewide Planning and Research Cooperative System (SPARCS) database from 2005-2018, multivariate logistics models were used to predict endpoints including POAF, the Society of Cardiothoracic surgeon (STS) endpoint (MM), and 30-day readmission (READMIT) in patients with and without mental illness. The TAVR procedure was approved for high-risk patients after 2012, and intermediate-risk patients after 2016, indicting a need to analyze the two populations separately. Multivariate analysis was only conducted on the first-time TAVR patients because of the small n in the viv-TAVR population. RESULTS/ANTICIPATED RESULTS: After 2012, 13.05% (1,810/13,870) of patients undergoing TAVR and 20.83% (15/72) undergoing viv-TAVR were diagnosed with a mental illness before the procedure. After 2016, 15.59% (1,485/9,524) TAVR patients and 20.00% (11/55) viv-TAVR patients had a preoperative diagnosis of mental illness. Multivariate analysis showed that mentally ill patients did not have significant differences in rates of POAF, 30-day readmission, and 30-day composite outcomes when compared to patients without mental illnesses following TAVR procedures after 2012 and 2016. Patients with POAF after both 2012 and 2016 were significantly less likely to be mentally ill, Black, and Hispanic. DISCUSSION/SIGNIFICANCE: Of the mentally ill patients who underwent TAVR, there was no significant difference in short-term outcomes after 2012 vs. 2016, compared to patients without mental illnesses. The small number of mentally ill patients undergoing TAVR may point to provider bias as a contributor to this high selectivity, and further evaluation would be of clinical use.
319 Optimizing Urinary cell mRNA profiling of kidney allograft recipients: Development of a home processing protocol for noninvasive diagnosis of T cell mediated rejection and BK virus nephropathy
Authors: Salinas Thalia , Li Carol , Snopkowski Catherine , Stryjniak Gabriel , Shankaranarayanan Divya , Albakry Shady , Ding Ruchuang , Sharma Vijay K. , Salvatore Steven P. , Seshan Surya V. , Dadhania Darshana M. , Muthukumar Thangamani , Suthanthiran Manikkam
Published: April 2023

OBJECTIVES/GOALS: Development of a user friendly home kit that enables kidney transplant recipients to process urine at home and post the lysate containing RNA to a Core Laboratory would simplify urinary cell mRNA profiling and facilitate longitudinal monitoring. We report our home processing protocol and investigation of its diagnostic performance characteristics. METHODS/STUDY POPULATION: We developed a home processing protocol (HPP) consisting of urine filtration and lysis of urinary cells, both performed at home by the kidney transplant recipients (KTR) themselves, followed by isolation of total RNA from the lysate and mRNA enrichment using a silica-membrane-based cartridge, both performed at the Core Laboratory. Using the HPP, total RNA was isolated from kidney allograft biopsy-matched urines and absolute copy numbers of CD3ÎµmRNA, CXCL10 mRNA, and 18S rRNA, components of the Clinical Trials in Organ Transplantation 04 (CTOT-04) three-gene TCMR diagnostic signature, and urinary cell BKV VP1 mRNA copy number, were measured using customized RT-qPCR assays. RESULTS/ANTICIPATED RESULTS: CTOT-04 three-gene TCMR diagnostic signature scores in urine processed using HPP discriminated KTR with TCMR (12 TCMR biopsies from 11 KTR) from KTR with no TCMR/BKVN (29 No TCMR/No BKVN biopsies from 29 KTR) (P=0.0005, Mann-Whitney test), and AUROC was 0.84 (95% CI, 0.69 to 0.98). TCMR was diagnosed with sensitivity of 67% (95% CI, 35 to 89) at a specificity of 86% (95% CI, 67 to 95) using the CTOT-04 validated cutpoint of -1.213 (P=0.0016,Fisher exact test). BKV VP1 mRNA copy number in urine processed with HPP discriminated KTR with BKVN (n=7) from KTR with no TCMR/BKVN (n=29) and AUROC was 1.0 (95% CI, 1.00 to 1.00). BKVN was diagnosed with a sensitivity of 86% (95% CI, 42 to 99) at specificity of 100% (95% CI, 85 to 100) with the previously validated cutpoint of 6.5x10^8 BKV VP1 mRNA copies/μg of RNA (P DISCUSSION/SIGNIFICANCE: Urine processed using the HPP predicted TCMR and BKVN in KTR. The HPP represents not only a significant advance towards portability of urinary cell mRNA profiling but also should improve patient management by minimizing visits for urine collection.