bluuebunny/crossref_metadata_2025 huggingface dataset:
320 Oxygenated Peritoneal Perfluorodecalin Improves Response to Normobaric Hypoxic Exposure in Swine
Authors: Leibowitz Joshua L , Awad Morcos A. , Stachnik Stephen , Moon Yejin , Kadkhodaeielyaderani Behzad , Hahn Jin-Oh , Fathy Hosam K. , Stewart Shelby J. , Friedberg Joseph S.
Published: April 2023

OBJECTIVES/GOALS: Patients suffering from respiratory failure have few options to support oxygenation and carbon dioxide removal aside from mechanical ventilation. Our objective was to test a novel extrapulmonary mechanism of gas exchange via peritoneal oxygenated perfluorocarbon (PFC) in a large animal model. METHODS/STUDY POPULATION: Using two 50 kg swine, hypoxia was modeled with subatmospheric oxygen and hypercarbia induced with acute hypoventilation. Through a midline laparotomy, cannulas were placed into the peritoneal space to allow for PFC infusion and circulation. After abdominal closure, these cannulas were connected to a device capable of draining, oxygenating, and infusing PFC. One animal was subjected to acute hypoxia (12% FiO2) and another animal to acute hypoventilation (4 breaths per minute). Primary outcomes were times for SpO2 to reach 75 mmHg, respectively. Trials were performed without PFC and with PFC dwelling or circulating through the peritoneal space, during which abdominal and bladder pressures were monitored and maintained under 20 mmHg by regulation of the PFC volume contained in the animal. RESULTS/ANTICIPATED RESULTS: In the animal subjected to acute hypoxia (12% FiO2), survival time improved from 5:55 to 20:00 (min:sec) after 2.5 liters of oxygenated PFC was instilled in the peritoneal space. Oxygen percent saturation of PFC before and after dwelling in the peritoneal space was measured at 100% before and 70% after dwelling in the animal during this hypoxic period corresponding with a gas transfer of 300 mL of oxygen over the 20-minute trial (i.e., 15 mL/min). Continual PFC circulation did not further extend the survival time during hypoxic conditions over PFC dwelling in the abdomen. In the animal that was acutely hypoventilated, there were no detectable differences in the rate of CO2 accumulation as measured by EtCO2 or direct blood pCO2 measurements with PFC dwelling or circulating through the peritoneal space. DISCUSSION/SIGNIFICANCE: Oxygenated PFC dwelling in the peritoneal space increased the duration of systemic arterial blood saturation remaining greater than 50% during normobaric hypoxic (12% FiO2) conditions but did not appreciably clear blood carbon dioxide during hypoventilation. Future experiments will focus on maximizing the rate of systemic oxygen uptake.
327 Radiopathomics: Integration of Advanced Neuroimaging and Molecular Pathology Features in Meningiomas
Authors: Haghdel Arsalan , Chang Se Jung , Ramakrishna Rohan , Magge Rajiv , Sabuncu Mert , Pannullo Susan , Schwartz Theodore , Knisely Jonathan , Pisapia David , Liechty Benjamin , Ivanidze Jana
Published: April 2023

OBJECTIVES/GOALS: Our overall objective is to investigate the relationship between radiologic features of meningioma with recently identified histopathological and molecular biomarkers, and to apply a machine learning (ML) approach to further demonstrate their utility in predicting clinical outcomes. METHODS/STUDY POPULATION: We have enrolled a cohort of 84 patients with meningioma diagnosed on the basis of conventional gadolinium-enhanced MRI imaging features since September 2019. Each patient has demographic and clinical data, Ga-68-DOTATATE MRI/PET SUV and dynamic metrics, DCE-MRI perfusion parameters, and histopathologic data. Various tumor subregions will be segmented semi-automatically and later confirmed by experienced neuroradiologist. Histopathologic data will include histologic grade, mitotic rate, Ki67 proliferative index, and presence of WHO established atypical histologic features, immunohistochemical parameters, and established high-grade molecular features. We will use supervised learning techniques to develop algorithms for predicting molecular features from imaging phenotypes. RESULTS/ANTICIPATED RESULTS: Anticipated results - advancements in understanding the molecular biomarkers of meningiomas has uncovered genetic alterations and epigenetic changes that more accurately determine tumor behavior. Currently, the imaging correlates of these molecular biomarkers are unknown, and utilizing radiographic data to predict prognosis and imaging-based classifications of meningiomas have not yet been investigated. Validated imaging correlates of molecular biomarkers not only provide an in-vivo assessment of tumor biology, but can also be integrated with histopathologic features ( radiopathomics models’) for more accurate disease prognostication. We anticipate that our results will identify surrogate imaging features for some of the recently emerged molecular biomarkers of meningioma. DISCUSSION/SIGNIFICANCE: There is a paucity of data on the importance of imaging phenotypes in determining tumor biology. This work has the potential of significant clinical impact by enabling a priori molecular characterization of meningiomas at the time of new diagnosis or recurrence, thereby allowing a personalized medicine approach to treatment planning.
328 Rapid Screening Tool for Identifying Acute Myocardial Injury: An Exploratory Study Evaluating the Ability of a Novel, Noninvasive Device to Detect Cardiac Troponin
Authors: McDonald Sam , Katz Jessie , Titus Jitto , Biswal Siddharth , Burman Atandra , Agusala Kartik , Diercks Deborah , Vigen Rebecca
Published: April 2023

OBJECTIVES/GOALS: Tropsensor, a noninvasive portable device using infrared spectroscopy, delivers a troponin res ult within five minutes, significantly quicker than standard of care (SOC) assays. This pilot study assesses the correlation of the Tropsensor and high sensitivity cardiac troponin (hs-cTnI) assay results. METHODS/STUDY POPULATION: Patients undergoing cTnI testing with the Abbott Architect STAT (Abbott Laboratories) hs-cTnI assay were recruited at a quaternary-care emergency department (ED). The Tropsensor was applied to the underside of the patients’ wrist within 5 minutes of the SOC blood draw for 5 minutes. The results of the hs-cTnI assays were compared with the raw output of the Tropsensor device to assess the relationship using both Spearman’s and nonlinear logarithmic measures of correlation. Patient demographic data was extracted from the EHR to supplement the data collected for this study. RESULTS/ANTICIPATED RESULTS: 58 patients were recruited with a mean age of 60 years (60% male, 40% female). Due to connection error, 8 patients’data did not get captured by the device. Additionally, due to noise related to suboptimal device contact with the wrist, 24 patients’data (41%) were rejected. Of the 26 patients with usable data, 9 patients had a troponin above the 99th percentile threshold. A nonlinear correlation of 0.64 and Spearman’s rho of 0.59 were observed between the SOC hs-cTnI assay and Tropsensor optical data. DISCUSSION/SIGNIFICANCE: The Tropsensor exhibits 64.4% correlation to the SOC hs-cTnI assay. While further evaluation is needed, this exploratory study provides insight into the potential of a transdermal optical device to be used as a screening tool for AMI in an ED triage situation.
332 Role of Kynurenine/Tryptophan Ratio in Kidney-Lung crosstalk in two porcine trauma-induced multi-organ injury models
Authors: Singh Pragya , Montemayor Daniel , Pamreddy Annapurna , Drel Viktor , Ye HongPing , Franzone Anthony , Zang Yanyi , Roberts Teryn , Chung Kevin , Batchinsky Andriy , Sharma Kumar
Published: April 2023

OBJECTIVES/GOALS: Multiple Organ Failure, often precipitated by Acute Lung Injury, is a life-threatening condition that causes death in military and civilian life. Furthermore, Acute Kidney Injury is very common, increasing morbidity and mortality rates. Therefore, understanding the molecular difference between survivors and non-survivors is urgently needed. METHODS/STUDY POPULATION: A 24-hour unilateral pulmonary contusion porcine model (pneumonectomy) of trauma-induced Multiple Organ Failure (MOF) model (n=17) and separate 48-hour polytrauma injury of bilateral pulmonary contusion, traumatic brain injury, and hemorrhage (polytrauma) MOF model (n=26) was developed at Dr. Batchinsky's AREVA laboratory. Serum was assayed at baseline and 3h or 6h post-trauma for amino acid metabolites using the Zip-Chip platform for mass spectrometry. The IDO1 enzyme activity assay kit (ab235936) was used to measure IDO1 enzyme activity in the tissue. Mass Spectrometry Imaging (MSI) was employed to frozen kidney tissues. Tissues were sectioned to 10- micron thickness. For MSI, the DAN matrix was utilized, and MALDI-MSI images were processed and obtained from METASPACE and SCILS software. RESULTS/ANTICIPATED RESULTS: In the pneumonectomy model, 10 survived, 7 died, and in the polytrauma group, 13 survived, and 13 died. In the pneumonectomy model, there was a significant increase in the serum kynurenine/tryptophan (KYN/TRP) ratio in the non-survivors 3h post-injury. A similar pattern was found in the validation group, which showed a significant increase in the KYN/TRP ratio at 6h post-trauma in non-survivors from the polytrauma model. There was a significant increase in IDO1 enzyme activity in non-survivor kidney tissues and a downregulation of tryptophan (TRP) metabolite in the kidney section in the non-survivor group post-trauma. DISCUSSION/SIGNIFICANCE: An increase in the KYN/TRP ratio post-trauma identified the pigs that suffered early mortality. A decrease in TRP metabolite and an increase in IDO1 enzyme activity in the kidney could contribute to an increase in KYN in the non-survivors. As a result, focusing on therapeutics targeting the KYN/TRP to reduce the incidence and severity of MOF is warranted.
334 Determinants of Health Affecting Self-Efficacy and Quality of Life in Patients with Prostate Cancer
Authors: Ohene-Agyei Jada A. , Hamilton-Reeves Jill , Camargo Juliana Teruel
Published: April 2023

OBJECTIVES/GOALS: Our long term goal is to identify the socioeconomic and cancer-specific determinants in patients with prostate cancer undergoing prostatectomy that impact their ability to maintain a healthy weight. This study explores the association between participants’health determinants and their indicated degree of self-efficacy and quality of life (QoL). METHODS/STUDY POPULATION: Study population includes participants of the WARRIOR trial (n=40), which included overweight men scheduled radical prostatectomy from the University of Kansas Medical Center. In addition to baseline demographics, the study team will administer a questionnaire based on a socioeconomic position and health-related QoL framework. This questionnaire will assess participants' socioeconomic, cancer-specific, and psychological circumstances at time of surgery and present day. Univariate analyses will be conducted on all variables with bivariate analyses between socioeconomic and clinical items to the outcome of composite self-efficacy scoring. RESULTS/ANTICIPATED RESULTS: We anticipate that participants in the intervention will report higher self-efficacy and emotional/social support than participants in the control group, participants with social vulnerability (lower income, marginalized race/ethnicity, etc) will report decreased self efficacy and poorer QoL compared with participants who are not socially vulnerable, and that participants who previously indicated social vulnerability will report more emotional barriers to weight loss, and lesser weight loss satisfaction, self-esteem, and QoL. DISCUSSION/SIGNIFICANCE: Lifestyle interventions have helped prostate cancer patients lose weight before surgery, but many regain weight. Exploring perceptions of self efficacy as well as learning more about what structural and systemic barriers affect self efficacy is important to inform how to improve our approach for sustained weight loss and health behavior changes.
337 Targeting metabolic and epigenetic programs to re-sensitize glioblastoma to chemotherapy
Authors: Rowland Emma , Walter Thomas , Suter Robert , Jermakowicz Anna , Riggins Rebecca , Ayad Nagi
Published: April 2023

OBJECTIVES/GOALS: Treatment options for glioblastoma (GBM) are limited. Prognosis remains dismal, with an 18 month on average survival rate following diagnosis due to treatment resistance and disease recurrence. The goal of this project is to investigate hallmarks of cancer progression that contribute to temozolomide (TMZ) resistance, a first tine treatment for GBM. METHODS/STUDY POPULATION: Two signaling pathways were investigated in TMZ-sensitive and -resistant GBM cell lines and in primary and recurrent patient-derived xenograft (PDX) tumor cells by genetically and pharmacologically inhibiting methionine adenosyltransferase 2A (MAT2A) and adenosylhomocysteinase (AHCY). Cell growth and survival were assessed by measuring protein expression of proliferation, oxidative stress and cell cycle arrest markers. EPIC array analysis and targeted bisulfite sequencing were conducted to identify changes in genome-wide and specific CpG island methylation. The Seahorse XF Analyzer measured mitochondrial respiratory capacity and oxidative metabolism. Induced pluripotent stem cell organoids were co-cultured with PDX tumor cells to determine if treatments mitigate tumor cell invasiveness. RESULTS/ANTICIPATED RESULTS: Compared to parental cells (PC), MAT2A gene expression was increased by 1.7-fold in acquired resistant and de novo resistant GBM cells (RC) [(transcript per million): PC, 7386 ± 0.012; RC, 12925 ± 0.023; n=2; p=2.10e-8]. Compared to TMZ-sensitive cells (TS), TMZ-resistant cells (TR) demonstrated a 56% increase in baseline oxygen consumption rate [(pmol/min): TS, 179 ± 6.7; TR, 279 ± 13; n=18; p=.0012] and 64% increase in maximal respiratory capacity [(pmol/min): TS, 403 ± 29; TR, 659 ± 35; n=6; p DISCUSSION/SIGNIFICANCE: MAT2A and AHCY contribute to TMZ resistance and recurrence by dysregulating methylation programs and upregulating antioxidant programs, respectively. These findings provide a foundation for developing novel combinatory therapeutic strategies and inform clinical studies intended to increase remission and reduce recurrence for GBM patients.
341 The Impact of Critical Social Determinants of Health on Personal Medical Decisions: Analysis of Older Americans in All of Us
Authors: Bhavnani Suresh K. , Zhang Weibin , Bao Daniel , Kuo Yong-Fang , Reistetter Timothy , Hatch Sandra , Urban Randy , Raji Mukaila , Downer Brian
Published: April 2023

OBJECTIVES/GOALS: A growing number of older adults in the United States have multiple social determinants of health (SDoH) that are barriers to effective medical care. We used generalizable machine learning methods to identify and visualize subtypes based on participant-reported SDoH profiles, and their association with delayed medical care (self-reported yes/no). METHODS/STUDY POPULATION: Data. All participants aged >=65 in All of Us with complete data on 18 SDoH self-reported variables, selected through consensus by 2 experienced health services researchers, and guided by Andersen’s behavioral model. Covariates included demographics, and the outcome was delayed medical care . Cases (n=4090) consisted of participants with at least one of the 18 SDoH variables, and controls (n=7414) consisted of participants with none of them. Method. (1) Used bipartite network analysis and modularity maximization to identify participant-SDoH biclusters, and visualize them through ExplodeLayout. (2) Used multivariable logistic regression (adjusted for demographics and corrected through Bonferroni) to measure the odds ratio (OR) of each participant bicluster to the outcome, compared with the controls. RESULTS/ANTICIPATED RESULTS: The analysis identified 7 SDoH subtypes (https://postimg.cc/Vd7Pg4xZ) with statistically significant modularity compared with 100 random permutations of the data (All of Us=.51, Random Mean=.38, z=20, P DISCUSSION/SIGNIFICANCE: The results identified 7 distinct subtypes based on SDoH profiles and their risk for delayed medical care, highlighting the importance of addressing specific combinations of barriers, with affordability having the highest risk. Furthermore, the analytical methods used are generalizable and have been made publicly available on CRAN and All of Us.
345 The Role of TCF7L2 in Hepatic Metabolic Zonation
Authors: Ayala Iriscilla I. , Hebbale Skanda K , Shannon Chris E. , Valdez Ivan , Fourcaudot Marcel , Bakewell Terry M. , Sholto Madelaine , Vallim Thomas , Heikkinen Sami
Published: April 2023

OBJECTIVES/GOALS: Single nucleotide polymorphisms in the transcription factor 7-like 2 (TCF7L2) gene are associated with Type 2 Diabetes (T2D) and nonalcoholic fatty liver disease (NAFLD). The metabolic function of TCF7L2 in the liver remains to be fully elucidated, but we hypothesized that TCF7L2 contributes to NAFLD through regulation of zonal metabolic pathways. METHODS/STUDY POPULATION: Using single nuclei RNA sequencing, we examined Tcf7l2 expression in periportal (PP) hepatocytes around the portal triad and pericentral (PC) hepatocytes surrounding the central vein of the liver. To visualize TCF7L2 transcriptional activity we used a TCF reporter mice, which expresses an H2B-eGFP fusion protein downstream of the conserved TCF DNA binding site. We disrupted Tcf7l2 transcriptional activity in mouse liver by breeding mice with a floxed Tcf7l2 exon 11, which encodes part of the DNA binding domain (DBD), to albumin-Cre mice (Hep-TCF7L2Î”DBD). Eight-week-old mice were fed a choline-deficient amino acid-defined high fat (CDAHFD) diet for 8 weeks. In liver samples harvested from these mice, we examined disruption to several key zonated metabolic pathways, and quantified the development of fibrosis. RESULTS/ANTICIPATED RESULTS: Single nuclei analysis revealed that Tcf7l2 mRNA was expressed primarily in parenchymal cells of the liver but was ubiquitous across the liver lobule. However, in immunofluorescence analysis of TCF reporter mice, the transcriptional activity of TCF7L2 was highly restricted to PC hepatocytes. Classic PC hepatocyte markers, including glutamine synthetase (Glul), were absent in Hep-TCF7L2Î”DBD mice. Following the CDAHFD, Hep-TCF7L2Î”DBD mice developed more severe fibrosis in histological analysis, and expressed elevated levels of genes involved in fibrogenesis, collagen synthesis and TGFβsignaling. Hep-TCF7L2Î”DBD mice also displayed hepatic cholesterol accumulation following the CDAHFD, which was likely the result of impaired pericentral bile acid synthesis. DISCUSSION/SIGNIFICANCE: Our results suggest that TCF7L2 plays an important role in the regulation of zonated metabolic pathways, which may contribute to the development of fibrosis. Ongoing analyses are exploring the mechanisms regulating the zonal transcriptional activity of TCF7L2.
360 Waste not, test more: Innovations in tissue processing to expand the testing of clinical specimens
Authors: Mojica Wilfrido , Yang Bei , Hsu Chang Chieh , Wu Yun , Izydorczak Alexandra , Wood Troy D. , Cho Dara , Parashurama Natesh , Yergeau Donald , Mahajan Supriya D
Published: April 2023

OBJECTIVES/GOALS: Clinical tissue specimens are primarily destined for formalin fixed, paraffin embedded processing to create a basis for diagnosis by microscopic examination. Innovations in specimen processing are required to expand its availability for inclusion as the substrate in assays that can contribute to the further development of Precision Medicine. METHODS/STUDY POPULATION: Transurethral resection of bladder tumors were selected for testing based on availability and tissue composition. A wash step was used to generate daughter aliquots composed of dislodged cells and a solution with prior contact to the parent tissue. This wash step served two purposes: 1) reduce the amount of contaminating material from spreading to other cases, a problem known to be associated with this type of specimen; and 2) create aliquots from which additional informative data could be generated. These daughter aliquots were then examined to determine their value as a source for exosome profiling, metabolomic studies, molecular characterization and organoid development. The parent tissue was not compromised, was able to undergo conventional processing and yielded results equivalent to unwashed specimens. RESULTS/ANTICIPATED RESULTS: Exosomes secreted by the tumor cells were identified to be present in the daughter aliquots by a combination of their isolation using CD31 and detection of miR-21 expression. These exosomes were confirmed to be not related to fragmented cells from testing for beta-tubulin. A global/discovery-based approach using mass spectrometry provided insights into early characterization of metabolomic profiles present in these tumor cells. Ample amounts of high quality DNA (226 ng/ul concentrations; 11.3 ug total) were recovered from the dislodged, excess cells in the wash for molecular studies. Finally, from viable cells recovered in one of the daughter wash aliquots, the ability to grow organoids was proven to be possible and reproducible. DISCUSSION/SIGNIFICANCE: Based on these results, the value of the clinical specimen can be markedly expanded for utilization in research and possible clinical use without detracting from the parent tissue. This non-destructive, easy to adopt wash procedure can potentially lead to an influx of data that may ultimately prove useful in improving patient care.
362 A Novel Animal Model of Radiation-Induced Heart Disease Using Photon Radiation
Authors: Whelan Emily , Di Florio Damian , Jain Angita , Hoppe Bradford , Fairweather DeLisa
Published: April 2023

OBJECTIVES/GOALS: The purpose of this study is to develop a clinically relevant mouse model of Radiation-Induced Heart Disease (RIHD) and characterize the resulting phenotype to find biomarkers and therapeutic targets as well as to understand the changes in cellular and molecular mechanisms of bioenergetics. METHODS/STUDY POPULATION: We used a two-beam method in the axillary region targeting the heart to irradiate male BALB/c mice at an isodose of 22, 16 and 8 Gray (Gy). We examined cardiac damage (i.e., vacuolization), inflammation, and DNA damage at 10 days post irradiation using histology and immunohistochemistry of heart tissue and cardiac function at day 35 by echocardiography. Additionally, cardiac tissue of mice irradiated at 22 Gy was collected at day 10 and day 35 post irradiation and sent for RNA sequencing. Data from RNA sequencing was analyzed using gProfiler, GSEA, and Cytoscape to enrich and visualize differentially expressed genes. RT-qPCR was performed to validate findings of significantly differentially expressed genes. RESULTS/ANTICIPATED RESULTS: Significantly increased phosphorylation of H2A.X indicated that irradiated mice were undergoing DNA double strand break repair indicating cardiac damage. Additionally, we found that regulators of mitochondrial function were decreased in the heart at day 10 for all doses. We found that mice that received 22 Gy developed cardiomyopathy at day 35 based on increased global longitudinal strain (GLS). Radiation decreased T cells, macrophages, and mast cells in the heart of irradiated mice by RT-qPCR at day 10 indicating damage to immune cells by radiation at all doses. Thus, we successfully created a clinically relevant model of RIHD in male BALB/c mice. DISCUSSION/SIGNIFICANCE: Patients undergoing radiation therapy for thoracic malignancies can develop cardiomyopathy (DCM) due to radiation damage. Previously published animal models utilized mouse strains resistant to developing DCM (female mice, C57BL/6 strain) and used high doses of radiation. Establishing a translational model is crucial for prevention of RIHD.
367 Designing an Untargeted Metabolomics Assay to Detect Biomarkers for Inborn Errors of Metabolism in the Clinical Laboratory
Authors: Wurth Rachel , Turgeon Coleman , ZinandrÃ©Stander  , Gavrilov Dimitar , Hall Patricia , Matern Dietrich , Schultz Matthew , Tortorelli Silvia , Oglesbee Devin
Published: April 2023

OBJECTIVES/GOALS: To develop an untargeted metabolomics assay that can holistically characterize the small molecule signatures of different inborn errors of metabolism (IEM) for biomarker discovery and identification of novel IEMs, with the goal of implementing the assay into the clinical laboratory to improve testing efficiency. METHODS/STUDY POPULATION: A hydrophilic interaction liquid chromatography (HILIC) column and reverse phase (RP) column were assembled in tandem on a SCIEX X500B quadrupole time-of-flight (QTOF) system to create a dual liquid chromatography (LC), tandem mass spectrometry method. The X500B was operated in data-independent acquisition mode with both positive and negative ionization. A mixture of 165 reference standards from eleven compound classes common to IEMs were used to evaluate the capability of the assay to resolve small molecules. Chromatographic resolution for each standard was determined qualitatively by comparison to a reference spectral database. External validation of the assay will be performed by analyzing a commercial library of reference metabolites. RESULTS/ANTICIPATED RESULTS: A total of 88% (146/165) of the standards were detected by the assay. The RP column alone resolved 71% (117/165) of the standards, the HILIC column resolved 33% (55/165), while 17% (29/165) of the standards were resolved by both columns. The HILIC column resolved standards that were more polar, while the RP column resolved more non-polar compounds. To evaluate matrix effects, the reference standard mixture was spiked into pooled plasma. In the presence of plasma 6/146 (4%) of the standards were suppressed to levels below the limit of detection. We expect external validation with the commercial metabolite library will corroborate these results, and that the high-quality spectral data attained from this reference library can be used to improve identification of unknown metabolites in patient samples. DISCUSSION/SIGNIFICANCE: We have shown our untargeted metabolomics assay can detect known biomarkers for IEMs. Clinical implementation of this method could streamline diagnosis of IEMs while simultaneously improving patient outcomes by leveraging the metabolome for biomarker discovery, and improved understanding of IEM mechanisms to inform novel treatment strategies.
371 Fractalkine isoforms using gene therapy differentially regulate microglia activation and vascular damage in the diabetic retina
Authors: Rodriguez Derek , Church Kaira A. , Pietramale Alicia N. , Cardona Sandra M. , Vanegas Difernando , Muzzio Isabel A. , Nash Kevin R. , Cardona Astrid E.
Published: April 2023

OBJECTIVES/GOALS: Retinal inflammation caused by the activation of resident macrophages (microglia) during diabetes exacerbates glial cell dysfunction, resulting in neuronal loss. The goal is to use rAAV gene therapy to deliver neuronal-derived fractalkine (FKN), minimizing inflammation and vascular damage in the diabetic retina. METHODS/STUDY POPULATION: The human microglial receptor (CX3CR1) binds to FKN, a protein that is expressed on neuronal membranes (mFKN), and undergoes constitutive cleavage to release a soluble domain (sFKN). Deficiencies in CX3CR1 or FKN showed increased microglial activation and elevated retinal pathology. To understand the mechanism by which mFKN and sFKN regulate microglia function, recombinant adeno-associated viruses (rAAVs) expressing mFKN or sFKN were delivered to intact retinas during diabetes. Markers of neuronal loss, vascular damage, and inflammation were analyzed. We hypothesize that the administration of rAAV-sFKN but not rAAV-mFKN will prevent vascular and neuronal damage, and improve visual function. RESULTS/ANTICIPATED RESULTS: rAAV-sFKN minimized microglial activation, blood vessel rupture, fibrinogen deposition, and prevented neuronal loss, compared to mice treated with rAAV-mFKN in a mouse model of diabetic retinopathy (DR). rAAV-sFKN treated mice showed improved visual acuity using a two-choice discrimination task through learning-based behavior. rAAV-sFKN treatment correlated with the success rate of the mice finding the reward based on their ability to distinguish visual cues. Future studies will test the effects of rAAV-sFKN and rAAV-mFKN on microglia inflammatory cytokine release, optic nerve damage and synaptic neurotransmission, peripheral immune responses, and transcriptomic changes in microglia during diabetes. DISCUSSION/SIGNIFICANCE: Current therapies for DR are ineffective in restoring vision. rAAVs-sFKN delivery appears to act as a neuroprotective approach in the diabetic retina. sFKN serves as an alternative pathway to implement translational and therapeutic approaches, minimizing pathology and improving visual function.
381 The role of leucine-rich PPR motif-containing protein (LRPPRC) in myelin lipid metabolism
Authors: Palacios Bridgitte , Ren Jiangong , Hu Jian
Published: April 2023

OBJECTIVES/GOALS: Leigh Syndrome, French Canadian-Type (LSFC) is a neurometabolic disorder caused by mutation of mitochondria-related gene, LRPPRC. White matter lesions and demyelination in central nervous system are common in LSFC. LRPPRC is enriched in myelinating glial cells, yet its role is not known. Our goal is to elucidate its mechanistic role in myelination. METHODS/STUDY POPULATION: We crossed C57BL/6N mice bearing a LRPPRC-loxP allele with mice bearing a Plp-CreERT2 allele. Mice with the Plp-CreERT2 allele expresses a tamoxifen-inducible Cre under the control of the Plp promoter, which drives expression in oligodendrocytes. Using these strains, we can target the deletion of LRPPRC, via tamoxifen injection, in both newly formed myelin and mature myelin. Plp-CreERT2; LRPPRCL/L (LRPPRC-KO) or control littermate mice will be injected for LRPPRC deletion at developmental and maturation stages of myelin. Immunofluorescence and electron microscopy of isolated brain tissues will be used for myelin integrity analysis. Cognitive functions of the mice will be measured via behavioral tests. Lastly, we will submit tissues for lipidomic analyses to observe any lipid metabolite variation. RESULTS/ANTICIPATED RESULTS: Behavioral and motor defects would be expected in LRPPRC-KO mice performing in cognitive function tasks across myelin maturation stages. Electron microscopy-based structure analysis of optic nerve, corpus callosum, and spinal cord should reveal thin or loss of myelin on the axons of LRPPRC-KO compared to control. Immunofluorescence staining of major myelin structural proteins, including myelin proteolipid protein (PLP), myelin basic protein (MBP), and myelin-associated glycoprotein (MAG) would be expected have lower levels in LRPPRC deficient tissues. Since myelin is a lipid-rich species, we would also expect lipid concentrations to be affected. LRPPRC-KO lipidomic analyses of myelin-related lipids should depict lower levels in comparison to control, which would imply dysfunctional lipid metabolism. DISCUSSION/SIGNIFICANCE: There are limited studies in ameliorating neural deficits caused by LS and LSFC. Successful completion of this project would help elucidate the functions of LRPPRC in myelination and lipid metabolism and potentially provide insights for developing novel therapeutic strategies for alleviating the demyelination and neural deficits in LSFC.
385 Combining Cannabidiol with Prolonged Exposure Therapy for PTSD: Design and Methodology of a Pilot Randomized Clinical Trial
Authors: Straud Casey , Roache John , Ginsburg Bret , Baig Rais , King Van , Young-McCaughan Stacey , Peterson Alan
Published: April 2023

OBJECTIVES/GOALS: There is increasing evidence that cannabidiol (CBD) has promising potential to treat PTSD. However, more research is warranted to fully understand the benefits of CBD for PTSD. This poster will describe the design and methodology of one of the first ever pilot RCTs examining CBD (vs. placebo) combined with prolonged exposure therapy for PTSD. METHODS/STUDY POPULATION: This study is an early Phase II double-blind, pilot RCT. Participants are 24 individuals 18-65 years old who meet DSM-5 criteria for PTSD on the CAPS-5 and were recruited from local hospitals and the community. Individuals complete a standardized baseline assessment with an independent evaluator to assess study eligibility. Participants who meet study inclusion are randomized to 18 days of CBD 250mg (BID) or placebo delivered in combination with 10-sessions Prolonged Exposure (PE) psychotherapy over 2 weeks. Individuals begin medication 3 days prior to beginning PE to ensure steady state. Participants complete self-report and biomarker outcomes at select timepoints during study participation, and are also asked to complete a 1-month follow-up assessment following treatment. RESULTS/ANTICIPATED RESULTS: This aims of this study are to: 1) examine the safety, feasibility, and PTSD symptom reductions associated with the combined intervention; 2) evaluate biomarkers associated with the endocannabinoid system and stress response; 3) determine the association between changes in biomarkers and PTSD symptoms following treatment. It is expected that CBD+PE will be safe and feasible, and that there will be a detectable signal of CBD vs. placebo in the reduction of PTSD symptoms. It is also anticipated that CBD will have higher levels of endocannabinoids and lower stress response levels compared to placebo. Lastly, we expect that greater changes in biomarkers will be associated with lower levels of PTSD severity following treatment. DISCUSSION/SIGNIFICANCE: Although there is growing interest in cannabinoids for psychiatric conditions, such as PTSD, controlled trials are limited and have yet to examine the proposed intervention for PTSD. If successful, this study will enhance the feasibility of a larger, adequately powered RCT to address immediate and long-term improvements for PTSD treatments.
394 Examining the Landscape of Clinical Trials Targeting Alcohol or Opioid Use Among Homeless Individuals
Authors: Baltazar Bruno , Pacifici Eunjoo
Published: April 2023

OBJECTIVES/GOALS: To understand the current landscape of clinical trials involving the homeless population by examining opioid or alcohol use disorders and the challenges in clinical trial recruitment. METHODS/STUDY POPULATION: Clinicaltrials.gov was searched with the keywords homeless or unhoused. The search was limited to studies conducted in the United States that were recruiting, not yet recruiting, active and not recruiting, completed, and enrolling. The search findings were further characterized and categorized by the definitions that were used for homelessness. Next, the trials were grouped based on the inclusion of alcohol or opioid use: (A) had no relevant mention, (B) included alcohol or opioid use as a secondary or other outcome measure, or (C) alcohol or opioid were the primary focus of the trial. Lastly, patterns and trends were identified for these trials. RESULTS/ANTICIPATED RESULTS: Out of 161 trials, 77 trials that met search criteria were identified then grouped based on how they classified homelessness: McKinney-Vento (n=5, 6%), DHHS (n=4, 5%), HUD (n=2, 3%), HEARTH Act (n=3, 4%), Custom (trials that specified parameters for homelessness, n=12, 16%), Not Specified (trials that provided no parameters for homelessness, n=26, 34%), and Other/Ambiguous (trials that used enrollment in an independent program as parameters or had unclear parameters, n=25, 32%). Of the 77 clinical trials that targeted homeless populations, 65% did not include alcohol use and 100% did not include opioid use in any outcome measure, 22% included alcohol use in a non-primary outcome measure, and 13% included alcohol use as the primary outcome measure of the study. DISCUSSION/SIGNIFICANCE: The number of clinical trials targeting homeless populations has increased over time, yet there is still no universal definition for classifying an individual as homeless. This lack of harmonization poses a challenge when coupled with the findings that there is a lack of clinical trials targeting opioid or alcohol use disorders.
399 The development of a digital game to prevent suicide in youth who misuse substances
Authors: Fernandes Claudia-Santi F. , Giannattasio Francesca , Truong-Phan Trucy , Blumberg Hilary , Fiellin Lynn E.
Published: April 2023

OBJECTIVES/GOALS: Suicide is the second leading cause of death in teens. Substance misuse, including opioids, is a risk factor for suicidal thoughts and behaviors among teens. School-based approaches are promising, yet they lack evidence on their effectiveness. To fill this gap, we created supportED, a digital game to prevent suicide in youth who misuse substances. METHODS/STUDY POPULATION: We conducted focus groups and interviews to inform the development of the digital game and discuss approaches to enhance user experiences, acceptability, and feasibility of the intervention. A semi-structured focus group/interview guide was developed, pilot tested, and used in focus groups/interviews. The guides aligned with constructs from a safety planning intervention, a well-recognized, evidence-based suicide prevention intervention, to identify potential warning signs of emotional distress, coping strategies, and resources for seeking help to prevent suicidal thoughts and behaviors. Focus groups/interviews were led by a facilitator, who is also a certified school counselor and licensed professional counselor, and a co-facilitator who took field notes and provided debrief summaries. RESULTS/ANTICIPATED RESULTS: A total of 35 participants, including 10 high-school-aged teens [aged 16-19], 10 college-aged youth [aged 18-22], 10 school-based providers, and five interviews with adults who had experience with opioids in their youth informed the development stage. Of the focus group participants, 25.7% identify as female (n=23), 71.4% identify as male (n=6), and 2.9% do not identify as male or female (n=1); 60% identify as White (n=18), 20% identify as multi-racial (n=6), 10% identify as Black (n=3), 3.3% identify as Asian (n=1), and 6.7% identify as race unknown (n=2); and, 37.1% (n=13) identify as Hispanic/Latinx. Salient themes that emerged focused on opioids, substance misuse, mental health, bullying, grief, family, identity, and stigma to create six distinct narratives in the digital game. DISCUSSION/SIGNIFICANCE: Findings informed the development of a digital game to prevent suicide among adolescents who misuse substances. The digital game is currently being evaluated in a pilot RCT to assess the preliminary efficacy of the intervention, user experience, usability, and feasibility of delivering the intervention and protocols among 60 youth.
400 Exploring the Diversity of Expanded Access Patients at Michigan Medicine
Authors: Gravelin Misty , Frank Chelsea , Rigan Laurie , Wright Jeanne , Weatherwax Kevin
Published: April 2023

OBJECTIVES/GOALS: Lack of diversity within clinical trials is well known, but there is little data on the use of investigational products through other pathways, such as expanded access. This project sought to determine the demographic diversity of patients benefiting from expanded access at Michigan Medicine. METHODS/STUDY POPULATION: Previous quality improvement reviews provided the list of 271 patients for whom a single-patient, expanded access request had been approved by the FDA and University of Michigan IRBMED between 2005 and 2021. Demographic information was collected through the EMERSE tool, including age, legal sex, race, ethnicity, and zip code. These data were cross-referenced with available regulatory documentation on product requested, treatment area, and date of request. Descriptive statistics were performed using Excel. RESULTS/ANTICIPATED RESULTS: Patients who were approved to use an investigational product through expanded access at Michigan Medicine showed a wide geographic distribution, including 48 Michigan counties, 20 states, and 1 province. All age groups were served, with those between 30-49 underrepresented and those under 10 and over 60 overrepresented. Race data generally followed the proportions of the Michigan state census, including 76% white and 14% black or African American (expected: 79% and 14%) and 48% female (expected: 50%). On further breakdown, populations differed by specialty and county. DISCUSSION/SIGNIFICANCE: The distribution of Michigan Medicine patients with approved expanded access requests was similar to the population of Michigan with respect to age, sex, and race. Further research is needed to determine if this reflects equitable use or if these results are generalizable to other institutions.
404 If You Build It, Will They Come? Navigating Research Resources at CTSA Hubs
Authors: McClave-Liu Michelle , Purcell Mary , Abedin Zainab , DiMaria Elisabeth , Muhammad Kawthar , Pope Leah , Rincón Helena , Pincus Harold , Reilly Muredach
Published: April 2023

OBJECTIVES/GOALS: CTSA Program hubs provide a wide range of research support services (funding, training, consultations, etc.) to individuals and teams. The CTSA Program hub at Columbia University seeks to identify best practices across CTSA hubs in how they facilitate researchers to identify, navigate, and access services at complex academic medical centers. METHODS/STUDY POPULATION: o A landscape analysis across CTSA Program hub websites was conducted during December 2021-February 2022, with the goal of assessing the availability of research navigation services and the ease of accessing and requesting research support services at each hub. Websites of 66 CTSA hubs were accessed and browsed for the following: 1) if a research navigation or concierge service was available; 2) how to request and apply to use common services such as pilot funding, biostatistics, clinical research services; 3) if there was a contact form and/or email address for general inquiries. Binary coding (1=Yes, 2=No) was used to track and summarize if these features were available, and then further classification and observations were noted into the full data set. RESULTS/ANTICIPATED RESULTS: The landscape analysis determined that 68% of CTSA hubs offer a form of research navigation including consultative models (personalized guidance, studios) and web-based models (self-service web portals, graphics, toolkits). Consultative models could be classified into three levels of support ranging from general information sharing to providing scientific expertise to convening more intensive studio sessions. 92% of CTSA hubs have at least 1 system in place for researchers to request services with a majority of hubs using a mix of tools and systems. In addition, 36% of hubs have additional general contact forms and 75% have general email addresses to assist researchers in obtaining more information. DISCUSSION/SIGNIFICANCE: There is a relative lack of data and information on the effectiveness of different research navigation models across the CTSA network, and barriers for researchers to identify services remain (Elworth et al). Our team is planning additional evaluations including interviews with leaders at other CTSA hubs and researchers and trainees at Columbia.
407 Moving from observational studies to a clinical trial: the impact of obesity and surgical weight loss on breast imaging, tissue, and cancer screening experience: the B-BRITE study
Authors: Clelland Elle N , Gonzalez-Vargas Judy , Palilla Sarah G , Koliwad Suneil K , Greenwood Heather I , Schwartz Christopher J , Benz Chris C. , Alba Diana L , Mukhtar Rita A
Published: April 2023

OBJECTIVES/GOALS: Obesity is associated with increased incidence of breast cancer (BC), yet is not included in many lifetime-risk calculators. Obesity may impact breast cancer screening sensitivity. Retrospective studies show that bariatric surgery is associated with a lower risk of BC, but the effects of surgical weight loss on breast tissue are poorly understood. METHODS/STUDY POPULATION: We proposed a mixed-methods before and after study design to investigate the effects of surgical weight loss on breast tissue via pre- and post-weight loss breast tissue biopsies and imaging. In addition, we aimed to better understand barriers to BC screening for patients with obesity by conducting qualitative interviews. With institutional review board approval, we have begun recruiting 14 cisgender women who plan to undergo Roux-en-Y gastric bypass or sleeve gastrectomy. Participants must be at least 40 years old, with no prior history of breast biopsies or breast cancer and will undergo comprehensive breast cancer screening including mammography with quantitative density assessment, breast MRI, as well as breast core biopsies. RESULTS/ANTICIPATED RESULTS: We hypothesize that obesity and its associated metabolic changes lead to altered breast stroma, including increased inflammation, and tissue stiffness, with subsequent risk of carcinogenesis. If true, we expect to find obese women will have measurably increased inflammatory markers in their breast tissue, which are reduced after bariatric surgery. We expect that change in mammographic density may correlate with fibroglandular volume change on MRI; there are little data on change in background parenchymal enhancement in the setting of obesity and weight change and quantifying this will provide preliminary data for future work. Last, we expect that undergoing BC screening will be easier for patients after weight loss due to constraints of imaging equipment and potential bias in the screening process. DISCUSSION/SIGNIFICANCE: Screening for BC is paramount to improving outcomes yet people with obesity are screened less with worse outcomes. Studying the effects of weight loss on the breast may improve interpretation of breast imaging in the setting of obesity and identify markers of risk. Understanding barriers to screening may help us develop strategies to improve screening.
412 Utility of a Team Science and Project Management Approach to Providing Effective Participant Recruitment Support to Research Teams: The Indiana CTSI Recruitment Concierge Service (RCS)
Authors: Hudson Brenda L. , Claxton Gina , Anderson Angela , Delp Lynsey , Carmel J. Egan , Wiehe Sarah , Moe Sharon
Published: April 2023

OBJECTIVES/GOALS: Evaluate effectiveness of participant recruitment service to improve study enrollment through customer feedback surveys and recruitment data obtained by research teams utilizing services. Use survey information to conduct gap analysis of additional support needed for participant recruitment. METHODS/STUDY POPULATION: Participant enrollment is often cited as one of the most challenging aspects of clinical research. In 2021, the Indiana CTSI used project management techniques to design and pilot a Team Science approach for providing participant recruitment support to clinical research teams. This service called the Indiana CTSI Recruitment Concierge Service (RCS) is comprised of recruitment, community engagement, social media, communications, and project management expertise. Additional experts are chosen to participate based on the study needs (regulatory, population insight, informatics, clinical services, etc.) RCS customers are comprised of study teams from a variety of areas and research experience. These customers are sent surveys to evaluate the support they received and provide suggestions for improvement. RESULTS/ANTICIPATED RESULTS: The RCS assisted 72 study teams in 2021 and 85 (as of November) in 2022. These studies were referred via word of mouth as no advertising of the service had been done to date. All customers were provided a study specific consultation with recommendations of services and resources that would assist their study. Some services recommended were: local study listing and volunteer registry (All IN for Health), digital marketing support, materials design expertise, community engagement and healthcare patient recruitment guidance. The overall feedback from RCS customers has been positive with most teams indicating the support improved their study recruitment and/or engagement plan. RCS will use information obtained to develop a strategy for prioritizing services due to the overwhelming number of requests received. DISCUSSION/SIGNIFICANCE: Using project management techniques and a Team Science approach, the Indiana CTSI was able to develop a comprehensive participant recruitment service that integrates clinical research operations, community engagement, and informatics expertise to design study specific recruitment plans and coordination of services.