bluuebunny/crossref_metadata_2025 huggingface dataset:
286 Partnering and engaging in Indiana Communities through listening sessions and data sharing: successes of Connection IN Health
Authors: Savaiano Dennis , Wiehe Sarah , Claxton Gina , Hinshaw Karen
Published: April 2024

OBJECTIVES/GOALS: Participants will be able to identify how partnering with state health departments for a CTSA initiative assists in meeting shared goals by building infrastructure and sharing resources and list key components of a successful county engagement process that can be replicated. METHODS/STUDY POPULATION: Connections IN Health is a collaborative project with the Indiana Clinical and Translational Sciences Institute, Indiana Department of Health, Indiana University Simon Comprehensive Cancer Center, and community partners. We address chronic disease in Indiana with coalition-based engagement at the grassroots level, and partnering to provide technical assistance, resources, connections and evidence-based strategies to address the health challenges. Our methods of county engagement include surveying the broad community, conducting listening sessions with key community stakeholders, compiling, analyzing and sharing data, collaborating with existing local coalitions, assessing community readiness and suggesting evidence-based practices to implement. RESULTS/ANTICIPATED RESULTS: As a result of county engagement, local coalitions have current, local data to drive their efforts in improving local health rankings and outcomes. Community partners in eight counties have selected and implemented evidence-based strategies to manage and/or prevent diabetes, cardiovascular disease and/or stroke. Some examples include: starting a community garden with cooking classes in a food desert to provide fresh produce and nutrition/cooking knowledge, creating a mobile kitchen on hand for organizations to use to provide education and cooking skills, providing a passport for wellness at existing community events with food samples and health screenings and offering evidence-based educational programming such as Dining with Diabetes and Be Heart Smart. DISCUSSION/SIGNIFICANCE: Moving county health coalitions to evidence-based programming that has the greatest likelihood for success is a critical translational sciences challenge. Our rigorous and well defined approach yields significant improvement in local health coalition activities, sustaining their activities through long-term trusted relationships.
292 Activating community health workers: A community-academic partnership to understand vaccine hesitancy.
Authors: Gore Devyani , Stiehl Emily , Dworkin Mark , Peacock Nadine , Parsa Naseem , Martin Melissa , Chandler Cornelius , Ghebenei Diana , Hebert-Beirne Jennifer
Published: April 2024

OBJECTIVES/GOALS: In 2022, Chicago created the COVID-19 Response Corps, a cohort of community health workers (CHWs), trained to conduct contact tracing and vaccine outreach. Through an Earn and Learn program, corps members studied community-engaged participatory research, and co-led a rapid assessment with researchers to assess vaccine hesitancy in communities. METHODS/STUDY POPULATION: The Chicago COVID-19 Community Response corps worked to mitigate COVID-19 transmission in disadvantaged neighborhoods by activating CHWs, a diverse public health workforce from communities most affected by health and economic inequities. The Earn and Learn Program allotted 600 corps members up to 7.5 hrs/week of paid capacity building opportunities to learn new skills, pursue training programs, or college courses. Embodying a praxis of participatory action research and intergenerational organizing, corps members co-designed research questions and survey instruments, pilot tested the tools, trained other corps members on how to recruit and collect data, and contributed to the analysis and interpretation of the results. They generated evidenced-informed solutions to address future real-world problems. RESULTS/ANTICIPATED RESULTS: Corps members brought insight, cultural literacy, and lived experience that was invaluable in reaching the priority population of unvaccinated Chicagoans. They enhanced all aspects of the rapid assessment while conducting their work safely and comfortably in neighborhoods that outsiders consider challenging. Community member responses as to why they had not yet received a COVID-19 vaccine included being unable to risk putting what they saw as a rushed or improperly tested product into their bodies, to not being able to risk becoming ill even temporarily due to the potential for lost wages, as well as having other priorities in their lives which took precedence over concern about COVID-19, such as paying bills and feeding their families. DISCUSSION/SIGNIFICANCE: Research and evaluation benefits from the inclusion of CHWs. They are agile agents of change with the potential to replenish and repair trust in a fractured public health system. Engaging CHWs in evaluation work can strengthen community-academic partnerships and enhance the understanding of challenges and solutions to improving community health.
298 Improving AI Assessment of Cutaneous Chronic Graft-Versus-Host Disease using Unlabeled Patient Photographs
Authors: McNeil Andrew , Parks Kesley , Cowen Edward , Lehman Julia , Pichard Dominique , Shinohara Michi , Flowers Mary , Dawant Benoit , Tkaczyk Eric
Published: April 2024

OBJECTIVES/GOALS: Measuring the area of skin involvement in chronic graft-versus-host disease (cGVHD) relies on costly, time-consuming manual assessment, with high disagreement among experts (>20%). Our published AI method, trained on labeled 3D photos, showed promise for delineating affected areas. We aim to improve its performance using unlabeled 2D photos. METHODS/STUDY POPULATION: Our published AI model (baseline) was trained on 360 labeled photos of 36 cGVHD patients,from a 3D camera with calibrated distance and lighting.Our gold standard labels were contours around affected skin, marked by a trained expert. A second unlabeledcohort of 974 standard 2D photos of 8 cGVHD patients was used to improve the baseline model. First the baseline model predicted affected areas on the unlabeled photos. Photos with good predictions were added to the training set with their AI-predicted labels. The model was then re-trained with the expanded labeled set. Models were successively trained with more AI labels until performance stopped improving. AI performance was assessed on a test set of 20 photos from 20 patients unseen during training, labeled by 4 experts to improve accuracy. RESULTS/ANTICIPATED RESULTS: Model performance was calculated by comparing against the gold standard labels on the test set. To quantify the spatial overlap of labeled areas the Dice coefficient was used (0 is no overlap, 1 is complete agreement), where higher values are better. To estimate clinical error we used surface area error (Error), where lower values are better. On the test set, the baseline model had a median Dice of 0.57 [interquartile range: 0.39 – 0.82] and Error of 57.6% [20.2 – 103.3%]. Re-training with additional AI-predicted labels from 8 new patients, the model yielded a median Dice of 0.60 [0.35 – 0.80] and Error of 50% [12.5 – 103.8%]. This approach is being expanded to a further 300 unlabeled patients, where we anticipate significant improvements to AI performance and consistency. DISCUSSION/SIGNIFICANCE: Evaluating AI models in standard photos could provide a consistent method of assessing and tracking cutaneous cGVHD and relieve the burden of costly expert assessment. A reliable automated AI tool would provide a meaningful improvement to the current standard of manual assessment and could be easily applied to large patient cohorts.
299 A CTS team approach to assess the in vitro toxicity of microplastic fibers to human lung epithelial cells cultured at an air-liquid interface
Authors: O'Connor Amber , Shankar Sripriya Nannu , Lewis Anna , Ferguson Lee , Wu Chang-Yu , Attwood Tara-Sabo
Published: April 2024

OBJECTIVES/GOALS: Our goal is to determine whether microplastic fibers (MPFs) provide signals for dendritic cell-induced Th2 polarization via epithelial-cell-derived thymic stromal lymphopoietin (TSLP). We seek to highlight a potential mechanism for MPF-induced airway toxicity associated with asthma exacerbation. METHODS/STUDY POPULATION: Primary human bronchial epithelial cells (NHBEs) were grown and differentiated at an air-liquid interface. Dyed and undyed polyester MPFs (14x45 µm) generated using a cryomicrotome were delivered to NHBEs through a custom designed mesh-hopper system. After the exposure period (6, 12, 24 hrs), cell viability was assessed using alamarBlue, and RT-qPCR was performed to determine mRNA expression of asthma associated genes (i.e., TSLP, IL-13, IL-33, etc.,) in NHBEs. Bulk mRNA-sequencing followed by bioinformatics will be performed to observe other plausible pathways tweaked by lung cell exposure to MPFs. RESULTS/ANTICIPATED RESULTS: Through gravimetric analysis, it was determined that the mesh-hopper system can achieve delivery efficiencies of at least 85% for as low as 500 fibers. Following exposure, results show polyester MPFs (500 - 1,000 fibers) exposed to NHBEs at multiple time points (6, 12, 24 hrs) did not result in a statistically significant decrease in cell viability. Treatment with 500 undyed MPFs resulted in a slight increase in TSLP expression at 6 hrs that decreased over time, whereas all other treatment groups resulted in TSLP downregulation. Similarly, 500 undyed MPFs resulted in an increase in IL-13 expression at both 6 and 12 hrs with all other treatment groups leading to IL-13 downregulation. We anticipate the RNA-seq results will show pro-inflammatory pathways are highly targeted following NHBE exposure to MPFs. DISCUSSION/SIGNIFICANCE: This study is one of the first to mechanistically assess the impact of MPFs on lung cells while simultaneously addressing the need for a reliable system that delivers MPFs to ALI cultures to better mimic inhalation and avoid inadequate resuspension of particles in liquid medium.
302 Diagnostic Evidence Gauge of Spatial Transcriptomics (DEGAS-ST): Using transfer learning to map clinical data to spatial transcriptomics in prostate cancer
Authors: Couetil Justin , Liu Ziyu , Alomari Ahmed K. , Zhang Jie , Huang Kun , Johnson Travis S.
Published: April 2024

OBJECTIVES/GOALS: The 'field effect' is a concept in pathology that pre-malignant tissue changes forecast health. Spatial transcriptomics could detect these changes earlier than histopathology, suggesting new early cancer screening methods. Knowing how normal tissue damage relates to cancer’s origin and progression may improve long-term outcomes. METHODS/STUDY POPULATION: We trained DEGAS, our machine learning framework, with prostate cancer data, combining both general cancer patterns and in-depth genetic information from individual tumors. The Tumor Cancer Genome Atlas (TCGA) shows how gene patterns in tumors relate to patient outcomes, emphasizing the differences between tumors from different patients (intertumor). On the other hand, spatial transcriptomics (ST) shows the genetic variety within a single tumor (intratumor) but has limited samples, making it hard to know which genetic differences are important for treatment. DEGAS bridges these areas by finding tissue sections that resemble those in TCGA profiles and are key indicators of patient survival. DEGAS serves as a valuable tool for generating clinically-important hypotheses. RESULTS/ANTICIPATED RESULTS: DEGAS identified benign-appearing glands in a normal prostate as being highly associated with poor progression-free survival. These glands have transcriptional signatures similar to high-grade prostate cancer. We confirmed this finding in a separate prostate cancer ST dataset. By integrating single cell (SC) data we demonstrated that cells annotated as cancerous in the SC data map to regions of benign glands in the ST dataset. We pinpoint several genes, chiefly Microseminoprotein-β (MSMB, PSP94), where reduced expression is highly correlated with poor progression-free survival. Cell type specific differential expression analysis further revealed that loss of MSMB expression associated with poor outcomes occurs specifically in luminal epithelia, the putative progenitor of prostate cancer. DISCUSSION/SIGNIFICANCE: DEGAS reveals that normal-appearing tissue can be highly-associated with tumor progression and underscores the importance of the 'field effect' in cancer research. Traditional analysis may miss such nuance, hiding key transitional cell states. Validating gene markers could boost early cancer detection and understanding of metastasis.
303 Social Network Analysis of Patient Sharing Among Providers: Implications for Analyzing Disparities in Cancer Screening
Authors: Bhavnani Suresh K. , Zhang Weibin , Kuo Yong-Fang , Downer Brian , Reistetter Timothy , Hunter Rodney
Published: April 2024

OBJECTIVES/GOALS: Many providers share patients resulting in networks where clinical information is exchanged, and which can impact the quality and efficiency of care. Here we analyzed the network properties of a primary care service area (PCSA) in Harris County TX, motivating our ongoing analysis of how they are associated with disparities in cancer screening. METHODS/STUDY POPULATION: Data.All providers (n=731, Medicare 2018) from the PCSA with the most providers in Harris County TX, with gender, specialty, and the number of shared patients. Method. Modeled the data as a network consisting of provider nodes, connected in pairs by edges if they shared >11 patients (an empirically-determined threshold). Analyzed the network structure using (1) modularity maximization and its significance to identify densely-connected communities; (2) degree centralization to measure whether a few providers shared many patients, and betweenness centralization to measure whether a few providers connected densely-connected communities; and (3) chi-squared to measure if pairs of connected providers tended to be of the same gender compared to disconnected provider pairs. RESULTS/ANTICIPATED RESULTS: The results (Fig. 1, http://www.skbhavnani.com/DIVA/Images/Fig-1-SNA-Network.jpg [http://www.skbhavnani.com/DIVA/Images/Fig-1-SNA-Network.jpg]) revealed a fragmented network with 120 small components (connected subnetworks, not part of any larger connected subnetwork), and 1 large component. The large component (n=244) had strong and significant modularity (Q=0.73, z=53.13, P<.001) with communities of providers that shared more patients than expected by chance; low degree centralization (dc=0.11) suggesting that no provider dominated patient sharing, in addition to high and significant betweenness centralization (bc=0.5, P<.01) suggesting that a few providers were responsible for connecting the densely-connected communities; and a significant gender bias (X2=10.05, df=1, P< .01) among those that shared patients, versus those that did not. DISCUSSION/SIGNIFICANCE: The analysis revealed a specific type of vulnerability (betweenness) for network fragmentation, and a gender bias in how patients were shared. These results motivated our ongoing analysis on how the network properties are associated with disparity in cancer screening within PCSAs across Texas.
343 Development of osteoclast derived exosomes for vascular calcification therapy
Authors: Jennings Isabella , Zhou Hao , Kuncorojakti Suryo , Purwaningrum Medania , Scatena Marta , Giachelli Cecilia M.
Published: April 2024

OBJECTIVES/GOALS: The global incidence of calcific aortic valve disease (CAVD) increased 3.5-fold since 1990. No preventative or therapeutic pharmaceutical therapies exist for CAVD. We will establish the therapeutic potential of osteoclast-derived exosomes though characterization of contents and mechanisms of action to protect against mineralization. METHODS/STUDY POPULATION: Exosomes were purified from conditioned media collected from murine myeloid precursor cells, RAW264.7 (control), and osteoclasts induced to differentiate from RAW264.7 cells (OD). Protein content of exosomes was determined using proteomic analyses. Nucleic acid contents will be identified by sequencing mRNA, miRNA, and DNA. The calcification prevention and reabsorption abilities of control and OD exosomes will be tested using human valvular interstitial cells (VIC) and smooth muscle cell calcification assays and acellular osteologic disc assays, respectively. Comparison between cellular and acellular systems will help identify mechanisms of action, and demonstrate potential therapeutic viability of OD exosomes in preventative vs resorptive treatments. RESULTS/ANTICIPATED RESULTS: OD exosomes, but not control exosomes, prevented calcification in VIC in vitro. OD exosomes contained osteoclast-specific proteins including TRAP, MMP6, cathepsin K, and bone reabsorption factors including V type proton pumps, ATPases, and integrins. These genes are also involved in resorptive activities, and were highly upregulated in OD compared to control exosomes. We anticipate miRNA signatures associated with mineral resorption will also be present. Increased knowledge of exosome cargo will illuminate their mechanism of action and allow future work to engineer increased efficacy. We also anticipate a therapeutic response when OD exosomes are applied after calcification has begun, showing exosomes promote calcium reabsorption. DISCUSSION/SIGNIFICANCE: Establishing therapeutic potential and examining mechanisms of action will pave the way for OD exosomes as a CAVD treatment. Analysis of exosome contents will determine active molecules to be enhanced in future studies. This work will lay a foundation for moving into aortic valve organoid models, which are accepted by the FDA for preclinical trials.
344 Understanding drivers of post-Ebola syndrome (PES) in pediatric survivors of Ebolavirus disease: characterization and the way forward.
Authors: Bond Nell G , Engel Emily J. , Kanneh Lansana , Samuels Robert J. , Grant Donald S. , Schieffelin John S.
Published: April 2024

OBJECTIVES/GOALS: Ebolavirus disease survivors report persistent, debilitating health concerns dubbed Post-Ebola Syndrome (PES). Attention to PES in young survivors is lacking, we describe PES in pediatric EVD survivors in Eastern Sierra Leone. Additionally, we introduce our proposal investigating differential presentations of PES in pediatric survivors. METHODS/STUDY POPULATION: EVD survivors were enrolled a median of 2.5 years after resolution of disease. Survivors were eligible if listed in a national register maintained by the Sierra Leone Association of Ebola Survivors. Household contacts (HCs) were identified by survivors. Participants were assigned into three comparison groups: pediatric (7-11), adolescent (12-17) and young adult (18-25). A self-reported symptom questionnaire, and a physical exam were conducted. Variables were clustered within organ system and compared across groups. RESULTS/ANTICIPATED RESULTS: Pediatric survivors had lower levels of long-term sequelae compared to adolescents and young adults. Symptoms and abnormal physical exam signs increase with age. Musculoskeletal, psychiatric, ophthalmologic, and GI signs and symptoms were significantly different between groups. Pediatric survivors had significantly more persistent sequelae than age-matched HCs with no history of EVD; particularly within the cardiac/GI (p=.006) and psychiatric/neurological (p=.025) clusters. PES is heterogeneous with respect to age, calling for a deeper understanding of age-based differences. Even the youngest group of survivors experienced significantly more sequelae than HCs, highlighting the elevated symptom burden in these children over their peers. DISCUSSION/SIGNIFICANCE: Understanding mechanistic drivers will ultimately improve targeted treatments for PES. We will characterize symptom groups defining PES in children, determine the relationship between accelerated aging and PES in this population, and test how immune profiles associated with accelerated aging relate to the development of PES in children.
357 Effects of Dietary Nitrate Supplementation on Vascular Function in Individuals with Prediabetes
Authors: Arney Blaine E. , Fulton Timothy J. , Sundberg Christopher W. , Hunter Sandra K.
Published: April 2024

OBJECTIVES/GOALS: Impaired vascular function, a subclinical marker of cardiovascular disease, has been identified in prediabetes. Dietary nitrate supplementation has been shown to improve vascular function. However, this has not been studied in prediabetes. The purpose was to determine the effects of dietary nitrate on vascular function in prediabetes. METHODS/STUDY POPULATION: Five individuals with prediabetes (4 men, 1 woman; 55 ± 17 yr; HbA1c = 5.8 ± 0.2) participated in a double-blind, placebo-controlled, repeated measures study. Participants were randomly assigned to a 3-day nitrate supplementation (nitrate-rich beetroot juice, 12.9 mmol, 140 mL), or a placebo supplementation (nitrate-depleted beetroot juice, 0.05 mmol, 140 mL). Following supplementation, participants reported to the lab for measures of vascular function in the lower limb. Doppler ultrasonography was used to measure flow-mediated dilation (FMD) and post-occlusive reactive hyperemia (RH) of the superficial femoral artery in response to a 5-min bout of leg ischemia. RESULTS/ANTICIPATED RESULTS: FMD did not differ between the nitrate-rich (2.87 ± 2.01%) and placebo (2.24 ± 1.69%) conditions (p = 0.48; d = 0.35). Furthermore, peak RH did not differ between the nitrate-rich (1503 ± 443 ml/min) and placebo (1762 ± 414 ml/min) conditions (p = 0.36; d = 0.46). DISCUSSION/SIGNIFICANCE: These preliminary results suggest that dietary nitrate supplementation in the form of beetroot juice does not improve vascular function in individuals with prediabetes.
369 Investigating the impact of bariatric surgery on metabolic mechanisms that promote obesity-associated inflammation in subjects with and without Type 2 Diabetes
Authors: Hart Samantha , Steiner Joshua , Johnson Lance , Nikolajczyk Barbara
Published: April 2024

OBJECTIVES/GOALS: This project will provide novel insights into mechanism(s) by which differences in inflammation develop & resolve, or fail to resolve, in metabolically different groups of bariatric surgery patients determined by Type 2 Diabetes status. My work may uncover unique differences between cohorts, encouraging development of personalized medicine. METHODS/STUDY POPULATION: I analyzed human blood samples collected before and 3, 6, & 12 months after bariatric surgery at the University of Kentucky through an established tissue bank. Subjects hadnormal glucose tolerance, pre-diabetes, or Type 2 Diabetes, based on HbA1c%. Iisolated peripheral blood mono nuclear cells & will comparecytokine profiles among cohorts across all time points. I will define & perturbmetabolic differencesin immune cells among cohorts & across time via isotope tracing, fuel source limitation, and metabolite inhibition. This will determine causal relationships between cytokine profiles & immune cell metabolism. RESULTS/ANTICIPATED RESULTS: I anticipatecytokine profiles, a functional output of immune cells, will differ among cohorts pre-surgery, and that this difference will diminish post-surgery. Differencesmay be insignificant by the 12 month time point. I also anticipate differences in fuel usage and metabolite productionin immune cells among cohorts pre-surgery, and that these differences only partially resolve post-surgery to poise immune cells for continued chronic inflammatory action. I hypothesize that T2D status has a lasting impact on immune cell function and fuel usage patterns, and will continue to supportchronic inflammation following short term T2D remission and longer-term weight loss. DISCUSSION/SIGNIFICANCE: There has been an alarming increase in obesity and its comorbidities over recent decades, and inflammation is a known supporter of T2D. The anticipated rewiring of immune cell metabolism post-surgery, if incomplete, may poise subjects forweight regain and T2D recurrence.
370 Point-of-Service Salivary Microbiome Analysis in the Prevention and Detection of Oral Premalignant Lesions
Authors: Weikel Dana R. , Sultan Ahmed S. , Basile John , Ma Tao , Meiller Timothy F.
Published: April 2024

OBJECTIVES/GOALS: Our aim is to establish soluble salivary biomarkers indicative of increased risk of oral premalignancy to be used in a point-of-service technology. Our goal is to non-invasively assess risk level for premalignancy by characterizing a molecular signature pattern that can be applied to such a diagnostic tool at routine dental or medical visits. METHODS/STUDY POPULATION: Adult patients 18 years of age and older who are non-smokers and patients of the University of Maryland School of Dentistry Oral Medicine Clinic and have been diagnosed with oral premalignancy (proliferative verrucous leukoplakia) are eligible. Exclusion criteria include history of immunosuppression or immune compromise; use of antifungal, antibiotic, and/or antiviral medications within the past three months; and gross dental disease. Serial unstimulated saliva samples will be collected at baseline or diagnosis of oral premalignancy, 6 months and 12 months. Solubility testing will be completed to determine whether malignant markers such as EGFR/mTOR/PI3K/p53 are soluble in saliva, and patient samples will be analyzed by ELISA and compared to appropriate controls. RESULTS/ANTICIPATED RESULTS: We anticipate demonstrating increased activity of molecular pathways known to be involved in malignant transformation, such as EGFR/mTOR/PI3K/p53, or increased burden of select microbial pathogens to be associated with increased risk of oral premalignancy in the form of proliferative verrucous leukoplakia. Preliminary sensitivity and specificity testing of the identified markers will provide additional insight to the utility of a diagnostic tool with salivary specimen. Therefore, the microbiome and/or molecular profile proposed from these results will serve as a translational application to development of future point-of-service test devices to be used in the prevention and detection of oral premalignant lesions. DISCUSSION/SIGNIFICANCE: Oral cancer is the sixth most common cancer worldwide, and presents challenges in its diagnosis and clinical management. Later diagnosis is associated with poorer patient outcomes—therefore, a molecular and microbiome profile that may be used in a noninvasive diagnostic test technology would prove beneficial to providers and patients.
373 The brain under stress: How a history of unpredictable shock affects neural processing of future stressors
Authors: Martino Michael , Paniccia Jackie , Doncheck Beth , Clarke Rachel , Grant Ian , Ward Amy , Pagoota Bayleigh , Francis Megan , Smith Alex , Otis Jim
Published: April 2024

OBJECTIVES/GOALS: Determine how a history of unpredictable foot shock in mice affects brain wide patterns of neural activation to future stressors. Additionally, we aimed to characterize how the paraventricular nucleus of the thalamus (PVT) is involved in the fear sensitization process. METHODS/STUDY POPULATION: We used a mouse model of stress enhanced fear learning, where stressed mice are first subjected to a series of unpredictable foot shocks in a novel context while control mice undergo exposure to the novel context without experiencing foot shock. Mice are then left undisturbed for 28 days, following which they are exposed to a single foot shock in a novel context. Mice are tested in the second context 24 hours after single shock, and the amount of time spent frozen in the context provides a measure of fear sensitization. Whole brain patterns of activation during the second context test will be assessed via whole brain optical clearing with antibody staining of immediate early genes. The role of the PVT in fear sensitization will be characterized using chemogenetic approaches. RESULTS/ANTICIPATED RESULTS: Our preliminary results demonstrate that mice display enhanced fear acquisition long after the initial experience of unpredictable shocks. We anticipate to identify regions previously implicated in fear learning and novel regions not previously described through our brain clearing approach. In addition, we anticipate chemogenetic inhibition of the PVT will reduce freezing to an auditory cue associated with the shock in the second context but not to the context itself. DISCUSSION/SIGNIFICANCE: Our findings will provide a comprehensive view of how a history of unpredictable stress affects whole brain processing of subsequent stressful experiences, and describe the role of the PVT in cued fear sensitization.
374 Operationalization of a Translational Ethics Program
Authors: Farroni Jeffrey , McNamara Vicki , Smith Elise
Published: April 2024

OBJECTIVES/GOALS: Within our CTSA hub, greater emphasis is on understanding innovation and human health impact of translational science writ large rather than focus on translational research projects. Our program is restructured to reflect distinguishing ethical features of translational science which are complimentary to traditional research ethics issues. METHODS/STUDY POPULATION: This descriptive analysis depicts the development of our ethics program as an exemplar of how to integrate into the research enterprise of an academic health science center that engages in translational research. Our relational approach is predicated on the embodiment of ethical values by all who are involved in research committed to proactive dialog, team building, and collaboration. This translational research culture is facilitated by a multidisciplinary ethics team who are embedded throughout the translational research enterprise. RESULTS/ANTICIPATED RESULTS: Our program is integrated into a translational science enterprise within a CTSA hub in four areas: relational structure (from leadership team to community engagement), education (from trainees to the research community to the public), support (through ethics consultation with multiple touchpoints in the translational science pathway), and team science (from team on-boarding and communication to D&I research of team science interventions). We have developed a research agenda examining research ethics topics that increase quality, applicability and downstream social impact of research; understanding translational science through historical and science & technologies studies lenses, and ethnographic and mixed-method approaches to understanding team science and the science of team science. DISCUSSION/SIGNIFICANCE: The integration of a translational ethics program provides attention to traditional research ethics issues regarding study conduct and integrity but also transcends those concerns to focus on the translational science enterprise itself through relationships, cultivating trust, team science, DEIA, and social responsibility.
387 Pathogenicity of a CCDC6-RET Fusion in Malignant Peripheral Nerve Sheath Tumor (MPNST)
Authors: Chandrasekaran Sanjay , Chen Zhiguo , Le Lu
Published: April 2024

OBJECTIVES/GOALS: RETgene fusions in sarcoma are rare and their impact on pathogenicity is unknown. Malignant peripheral nerve sheath tumors (MPNST) are a deadly, genomically heterogenous soft tissue sarcoma rarely harboring targetable aberrations. We present a case of a CCDC6-RET fusion MPNST sensitive to RET-inhibitor therapy in a xenograft model. METHODS/STUDY POPULATION: Lung tumor tissue was obtained per an approved collection protocol from a 21yo male patient with a spontaneous MPNST harboring an inactivating mutation in NF-1 and a CCDC6-RET gene fusion detected by a commercially available sequencing panel (Signatera). To confirm pathogenicity of the RET fusion, fresh tumor tissue was engrafted into immunocompromised NSG mice in the anterior and posterior flanks, harvested at ~10 weeks, and re-transplanted into bilateral flanks. When tumor diameters reached 0.5-1cm (~4 weeks), mice were randomized into 3 groups (n=6/group) and treated with either vehicle (V) (PBS), the RET-specific inhibitor selpercatinib (S) (20mg/kg twice daily), or the multi-kinase inhibitor cabozantinib (C) (30mg/kg daily) by oral gavage. Mice were monitored weekly for weight and tumor size. RESULTS/ANTICIPATED RESULTS: 92% (33/36) of implanted tumors were evaluable for treatment response. Pre-treatment tumor volumes (mm3) across all three groups were similar (mean/Std Dev – V: 230/111, S: 271/132, C: 230/123). At day 7, tumor growth was significantly inhibited by S and C versus V (ANOVA p < 0.001, post-hoc Tukey’s V vs S p= 0.0178, V vs C p< 0.0001, S vs C p= 0.0005). V-treated tumors increased in volume by 60% while S reduced tumor volume by ~80% and C reduced tumor volume by ~20%. S and C treatments were tolerated well. and S improved survival with 100% of mice alive at day 63 vs 0% in V and C groups. 6 of the 12 implanted tumors treated with S, 50% increased in size after ~6-weeks following a >90% initial tumor reduction in tumor volume. Follow-on molecular studies in S-resistant tumors are ongoing. DISCUSSION/SIGNIFICANCE: Targetable genomic changes in MPNST, especially in RET, are infrequent and often considered stochastic. Our findings suggest that precision medicine approaches pairing genomic sequencing and in vivotesting of target gene pathogenicity may guide treatment planning and novel discovery for rare, difficult to treat sarcomas.
391 Value estimation of the Diabetes Prevention Program: How well does clinical trial-based cost-effectiveness apply to the real world?
Authors: Olchanski Natalia , Weidner Samuel B. , Cohen Joshua T. , Kent David M.
Published: April 2024

OBJECTIVES/GOALS: Many economic evaluations rely on clinical trial data that may not represent real world populations and intervention effectiveness. We compare risk and cost-effectiveness for the Diabetes Prevention Program (DPP) clinical trial cohort and a real world population eligible for the national DPP to assess the impact of using real world data. METHODS/STUDY POPULATION: To produce real world (US population) representative results, we identified National Health and Nutrition Examination Survey (NHANES) subjects eligible for the national DPP and adjusted projections using survey weights. We used clinical predictive models to estimate individual diabetes risk, and microsimulation to estimate lifetime costs, benefits, and net monetary benefits (NMB) for lifestyle intervention and metformin. We compared results across the DPP clinical trial and NHANES populations. RESULTS/ANTICIPATED RESULTS: Three-year risk of diabetes onset for the DPP trial population (mean of 19.7%, median of 10.3%) exceeded corresponding risk for the NHANES population (mean of 14.6%, median of 4.8%). The proportion of individuals with a three-year diabetes risk < 10% for the DPP trial population (49%) was less than the corresponding proportion for NHANES (67%). Mean NMB for metformin for the DPP trial population ($9,749) exceeded the corresponding value for NHANES ($5,391). The proportion of subjects with negative NMB was 49% for the DPP trial population and 67% for NHANES. Lifestyle intervention had a mean NMB of $34,889 for the DPP trial population and $28,652 for NHANES. Only 20% of the NHANES population eligible for national DPP met inclusion/exclusion criteria for the DPP trial. DISCUSSION/SIGNIFICANCE: Real world populations eligible for the national DPP include a greater proportion of low-risk individuals, and for these people, prevention programs may confer smaller benefits. Technology assessments based on clinical trial data should be revised using real world population and treatment effect data.
394 A Machine Learning Approach to Predicting High-Risk Irritability Trajectories Across the Transition to Adolescence
Authors: Jordan Leslie S. , Parker Alyssa J. , Wiggins Jillian Lee , Dougherty Lea R.
Published: April 2024

OBJECTIVES/GOALS: Irritability, a proneness to anger and frustration, is a transdiagnostic symptom associated with poor mental health outcomes. Levels of irritability vary across development and high-risk trajectories have been observed. This study aims to use machine learning to predict irritability trajectories across the transition to adolescence. METHODS/STUDY POPULATION: Data were from the Adolescent Brain Cognitive Development (ABCD) Study, which is a 10-year longitudinal study that tracks the brain development, cognitive skills, physical health, and psychosocial functioning of a large, national sample starting from preadolescence. The baseline sample consisted of 11,861 9-10-year-old preadolescent youth. Irritability was parent-rated at baseline, 1-year, 2-year, 3-year, and 4-year follow-ups on the Child Behavior Checklist (CBCL) irritability index. Latent class growth analysis (LCGA) was used to determine developmental trajectories of irritability. Two machine learning approaches were applied to develop predictive models of youth irritability developmental trajectories. We used baseline (preadolescent) variables that spanned a wide range of domains. RESULTS/ANTICIPATED RESULTS: Preliminary results fromthe LCGA indicated best support for a four-class model that differentiated growth trajectories in irritability across the transition to adolescence: 1) persistent low irritability (n = 8691, 73.27%), 2) moderate irritability and decreasing (n = 1257, 10.60%), 3) low to moderate irritability and increasing (n = 1295, 10.92%), and 4) chronic high irritability (n = 618, 5.21%). We expect the machine learning analyses to generate predictive models with acceptable accuracy. We hypothesize that the most important predictors in the models will originate from the youth mental health domain, including baseline youth irritability, externalizing symptoms, internalizing symptoms, and oppositional behaviors, and the parent psychopathology domain, particularly parent irritability. DISCUSSION/SIGNIFICANCE: The present study elucidates unique developmental trajectories of irritability and generates predictive models to classify high-risk irritability trajectories using machine learning approaches. Clinicians can use these predictive models to identify at-risk youth and provide early intervention to preadolescents at high risk.
400 Investigation of a translational astrocyte-targeted AAV-mediated gene addition therapy in two models of Vanishing White Matter disease
Authors: Herstine Jessica A. , Chang Pi-Kai , Chornyy Sergiy , Stevenson Tamara J. , Rediger Jessica , Wentz Julia , Pyne Nettie , Bonkowsky Joshua L. , Bradbury Allison M.
Published: April 2024

OBJECTIVES/GOALS: Vanishing White Matter Disease (VWM), is a childhood neurodegenerative leukodystrophy that presents with motor deficits, neurologic decline, and seizures leading to death.There are no treatments. Herein we investigate adeno-associated virus serotype 9 (AAV9) gene addition therapy for VWM. METHODS/STUDY POPULATION: To serve as a baseline for disease correction, we characterized the severe VWM Eif2b5I98M murine model with clinically relevant readouts including motor function, gait mapping and myelin loss through magnetic resonance imaging (MRI). Molecular characterization through the identification of biomarkers was also investigated. To provide targeted disease correction, we designed four gene replacement constructs to drive the rapeutic EIF2B5 expression in astrocytes—a critical cell type for VWM pathology. We are currently evaluating our AAV vectors in two murine VWM models, Eif2b5R191H and Eif2b5I98M, and are monitoring disease progression using traditional and clinically relevant readouts. RESULTS/ANTICIPATED RESULTS: The I98M mice display significant mobility loss, ataxic gait, and demyelination. Molecular characterization also indicates that the integrated stress response is significantly dysregulated, supporting the classic VWM phenotype. Our previous biodistribution study confirmed our ability to efficiently target astrocytes using varying iterations—including one novel—of the glial fibrillary acidic protein (GFAP) promoter. Our data suggests that targeting astrocytes with gene addition delays disease onset, partially rescues motor function, and attenuates myelin loss. Survival of the AAV9-gfaABC(1)D-EIF2B5 treated I98M mice is also significantly increased (p<0.0001), currently with a 2-fold extension in life expectancy. DISCUSSION/SIGNIFICANCE: Overall, we anticipate emergence of a lead astrocyte-targeted gene therapy candidate in which the data will be strengthened through the evaluation of clinically relevant measures in two murine models of disease, allowing fortimely translation to the clinic.
407 The OGT/O-GlcNAc Axis Regulates Fibrosis Resolution in Idiopathic Pulmonary Fibrosis
Authors: Vang Shia , Livelo Yiming , Burpee Bailey , Hirsch Meghan J. , Matthews Emma L. , Jones Luke I. , Melkani Girish , Krick Stefanie , Barnes Jarrod W.
Published: April 2024

OBJECTIVES/GOALS: Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by dysregulated collagen accumulation in the lung parenchyma. Our goal is to investigate the role of O-linked N-Acetylglucosamine (O-GlcNAc) transferase (OGT) in pulmonary fibrosis to ultimately discover novel therapies for fibrosis resolution. METHODS/STUDY POPULATION: Lung tissue from IPF and non-IPF donors was subjected to immunohistochemistry (IHC) to assess O-GlcNAc levels. Primary human lung fibroblasts were treated with OGT or O-GlcNAcase (OGA) inhibitors followed by transforming growth factor-beta 1 (TGF-β1) stimulation to assess O-GlcNAc regulation of fibroblast-to-myofibroblast transition (FMT) markers [alpha smooth muscle actin (α-SMA) and type 1 and type 3 collagen (COL1α1, COL3α1)] In Drosophila melanogaster, OGT knockdown (KD)/overexpression (OE) was conditionally induced to assess pericardin, a type IV collagen-like protein, regulation by immunofluorescence. Lastly, a mouse model of bleomycin-induced pulmonary fibrosis was examined following OGT KD and assessed for fibrosis resolution via histology, hydroxyproline assay, and western blotting. RESULTS/ANTICIPATED RESULTS: O-GlcNAc staining was increased in IPF lung tissue compared to non-IPF control lungs. In primary human lung fibroblasts, TGF-α1 administration resulted in increased FMT markers (α-SMA, COL1α1, and COL3α1), which were reduced or increased by OGT or OGA inhibition, respectively. Genetic manipulation in the Drosophila models showed decreased pericardin expression with OGT KD compared to the wild-type, whereas OGT OE increased pericardin compared to control. Additionally, OGT KD in bleomycin treated aged mice resulted in reduced collagen levels at the transcript and protein level and concurrent fibrosis resolution as assessed by Masson’s trichrome staining and total hydroxyproline analysis. Collectively, showing OGT/O-GlcNAc regulating collagen in fibrosis resolution. DISCUSSION/SIGNIFICANCE: These data suggest that the OGT/O-GlcNAc axis regulates collagen deposition in pulmonary fibrosis, and we show that O-GlcNAc is implicated in the pathogenesis of IPF. We identified OGT as a therapeutic target to overcome current drug limitations, opening new horizons for biomedical treatment.
410 Improving Patient Outcomes through Design of Biodegradable Implants for Long Bone Fractures
Authors: Unger Justin S. , Weihs Timothy P. , Guest James K.
Published: April 2024

OBJECTIVES/GOALS: Current long bone fracture standard of care uses inert metal intramedullary nails (IMN), 10x stiffer than femur cortex. Consequent “stress-shielding” bone loss sees >5% of patients needing revision surgery. To improve nonunion healing, we develop automated design optimization methods for biodegradable Mg alloy IMNs to control local reloading. METHODS/STUDY POPULATION: Finite element analysis (FEA) is performed on 3D bone-IMN representations to establish this study’s baseline strain states for existing inert IMN geometries within QCT-informed femoral models under simulated biomechanical loading. FEA with Mg alloy properties for same IMN designs simulate transient IMN material loss through discrete time-step models with experimental in vivo Mg corrosion rates and strain-based bone density evolution using remodeling algorithms from literature. Transient stability and strength metrics, fracture zone stress profiles under gradual reloading and manufacturing constraints are formulated through gradient-based sensitivity analysis into a topology optimization framework (TOF) incorporating a reaction-diffusion degradation model to generate IMN topologies. RESULTS/ANTICIPATED RESULTS: TOF designs for Mg alloy IMNs with transient allowable strength constraints, using safety factors to prevent IMN failure, demonstrate higher compliance than standard inert IMNs with mechanical properties closer to native cortical bone. The biodegradation model within the TOF, informed by corrosion behavior from bone-IMN FEA study, predicts how potential design evolutions affect transient strain states of the system. Thus, local fracture region stress states are controlled by the algorithm optimizing for desirable transient stiffness profiles based on a minimum variance objective of fracture zone stress compared to a target bone stress profile. Optimized IMNs with porous, high surface area features achieve 50% decrease in IMN stiffness over 6 months recovery time and complete in vivo degradation in 24 months. DISCUSSION/SIGNIFICANCE: Our TOF reduces “stress-shielding” effects via design for controlled IMN biodegradation to gradually increase fracture zone loading, stimulating remodeling and reducing current risk of post-operative fracture and surgical removal in ~15k cases/yr. in the U.S. In vitro mechanical and in vivo clinical testing is required to validate design results.
418 The antiplatelet effects of EPA, an omega-3 fatty acid, are mediated by its 12-lipoxygenase metabolite, 12-HEPE
Authors: Goerger Krista , Lee Taekyu , Gilmore Devin , Tran Michelle , Holman Theodore R. , Holinstat Michael
Published: April 2024

OBJECTIVES/GOALS: To determine whether cardioprotective effects observed in individuals taking dietary supplementation with eicosapentaenoic acid (EPA), an ω-3 polyunsaturated fatty acid, are realized by altering platelet function, and if these effects are mediated through the 12-lipoxygenase derived metabolite, 12-hydroxyeicosapentaenoic acid (12-HEPE). METHODS/STUDY POPULATION: Washed platelets or platelet rich plasma from healthy human donors were treated with EPA and 12-HEPE to assess their ability to inhibit platelet activation. Platelets were stimulated with agonists targeting different steps of the hemostatic response to vascular injury. Platelet aggregation, dense granule secretion, surface expression of integrin αIIbβ3 and P-selectin, and clot retraction were analyzed. To assess signaling through Gαs-GPCRs and protein kinase A activity, phosphorylation of vasodilator-stimulated phosphoprotein (VASP) was examined via western blot following treatment with EPA or 12-HEPE. RESULTS/ANTICIPATED RESULTS: EPA and 12-HEPE dose-dependently inhibit both collagen and thrombin-induced platelet aggregation. Furthermore, 12-HEPE more potently attenuates dense granule secretion and surface expression of platelet activation markers, integrin αIIbβ3 and P-selectin, in comparison to EPA. Plasma treated with EPA delayed thrombin-induced clot retraction, while 12-HEPE had no effect. Additionally, treatment with 12-HEPE increases phosphorylation of VASP, suggesting it could signal through the activation of the eicosanoid Gαs-GPCRs. DISCUSSION/SIGNIFICANCE: Here, we show for the first time that EPA directly inhibits platelet activation through its 12-LOX metabolite, 12-HEPE. These findings provide further insight into the mechanisms underlying the cardioprotective effects of EPA. A better understanding of current PUFA supplementations can inform treatment and prevention of cardiovascular diseases.