OBJECTIVES/GOALS: Achieving therapy for hepatocellular carcinoma (HCC) involves navigating through a complex cascade of care. Non-HCC cancer mortality has been associated with social determinants of health outside of cancer specific risk. Our objective is to explore the impact of social determinants on HCC outcomes. METHODS/STUDY POPULATION: Patients with HCC were enrolled from 3 hospitals form June, 1 2019 to December 1, 2019. A chart review was done to collect information on liver disease severity and cancer stage. Patients were interviewed to collect information on the following: 1) socioeconomic status (income, education, insurance status, and employment status), 2) literacy (Rapid Estimate of Adult Literacy in Medicine (REALM-R) and Brief Health Literacy Screening Tool (BREIF)), 3) social support (Patent Reported Outcome Measurement Information System (PROMIS) instrumental and information support tool), 4) quality of life (PROMIS global and mental health tool), 5) substance abuse, and 6) linkage to care. RESULTS/ANTICIPATED RESULTS: Data compiled on the social determinants of health revealed (n = 35): 1) 60.0% of patients had incomes below $30,000 per year, 60.0% of patients had not gone past high school for education, and 8.6% had full time employment, 2) the average BREIF score was 10.3 (range 3-15)(4-12 indicate limited literacy). The average REALM-R score was 5.5 (range 0-8) (<6 indicate at risk for poor literacy), 3) patients had strong instrumental (T score 61.4±7.1) and information social support (T score 64.6±4.7) (mean T scores calibrated to a general population mean of 50), 4) patients had poor mental (T score 43.7 ±6.5) and physical quality of life (T score 46.6 ±9.9), 5) 25.7% of patients reported alcohol use in the past 90 days 6) 80.0% of patients reported that their doctor had spoken to them about liver transplantation. DISCUSSION/SIGNIFICANCE OF IMPACT: This patient population was well linked to care with good social support. However their literacy, socioeconomic status, mental and global health was poor and substance use history complex. Continued follow up of this cohort is planned to determine how these factors might impact their ability to navigate through the care cascade as well as survival.

OBJECTIVES/GOALS: Pain Catastrophizing is a negative coping mechanism involving rumination, magnification, and helplessness and is associated with worse chronic pain. The neurobiological mechanisms underlying this relationship are poorly understood. We aim to examine the intrinsic activity of a functional pathway in patients with chronic orofacial pain. METHODS/STUDY POPULATION: This is the second phase of a parent study examining genetics, placebos, and the brain in temporomandibular disorders (TMD). We intend to recruit 120 of the original 398 TMD patients for this phase. Participants completed the Graded Chronic Pain Scale to assess TMD pain intensity and disability and the Pain Catastrophizing Scale. Behaviorally, pain catastrophizing scores and pain intensity and disability will be analyzed using structural equation modeling. Resting-state functional magnetic resonance imaging will be used to record intrinsic brain activity. The functional connectivity between the posterior cingulate, anterior insula, and periaqueductal grey will be assessed as a causal pathway relating pain catastrophizing to pain intensity and disability. Mediation analyses will be used to test causality. RESULTS/ANTICIPATED RESULTS: We anticipate that greater engagement in catastrophic thinking about pain increases the functional connectivity strength between the posterior cingulate, anterior insula, and periaqueductal grey, which ultimately leads to heightened perception of pain intensity and disability. Therefore, we expect to see increased functional connectivity in those with high pain catastrophizing levels as compared to those with low pain catastrophizing levels, and that this pathway will mediate the relationship between pain catastrophizing and pain intensity and disability. Further, we predict that helplessness will most strongly correlate with the change in functional connectivity as compared to rumination and magnification. Results will be presented in full at the conference. DISCUSSION/SIGNIFICANCE: Understanding how pain catastrophizing can influence chronic pain pathways will not only promote a more integrative approach to chronic pain management but will also help identify the mechanisms by which pain itself develops and persists in the particularly vulnerable pain population of TMD.

ABSTRACT IMPACT: Identification of microRNA (miRNA) associated with neuropathic pain after spinal cord injury (SCI) will elucidate underlying epigenetic mechanisms contributing to its development and identify targets for intervention to optimize treatment strategies and outcomes. OBJECTIVES/GOALS: Approximately 70% of individuals with SCI experience neuropathic pain, which is refractory to pharmacologic intervention, and can reduce overall health and wellbeing. This study aims to identify predictive miRNA biomarkers of neuropathic pain after SCI to identify targets for the development of efficacious interventions. METHODS/STUDY POPULATION: Blood samples and clinical outcome measures were collected from adult participants with SCI with neuropathic pain (n = 28) and without neuropathic pain (n = 15). The sample population consisted of a mean age of 39 (SD = 12.12), 8 female (20%), with 13 classified as acute SCI (within 3 months post injury) and 30 as chronic SCI (> 3 years post injury). Pain presence, type, and intensity were assessed with the International Spinal Cord Injury Basic Pain Dataset (ISCIBPDS). Serum miRNA sequencing counts were produced from blood samples. Fold change and independent t-tests assessed differential expression between those with and without neuropathic pain, and those with chronic or acute SCI. Linear regression was performed to explore the relationship between miRNA expression and ISCIBPDS pain intensity ratings. RESULTS/ANTICIPATED RESULTS: In individuals with SCI, significant downregulation of expression of miR-338-5p was present in those with neuropathic pain compared to those without neuropathic pain (fold change = 0.81, p = 0.04). A significant relationship between expression of miR-338-5p and highest reported neuropathic pain intensity on the ISCIBPDS was identified (R2 = 0.15, F = 7.32, p < 0.01). Covariates of sex, age, and years post injury were not found to significantly influence the relationship between miRNA expression and ISCIBPDS intensity ratings. No significant differences in miR-338-5p expression were identified between participants with acute and chronic SCI, or with nociceptive pain ratings, demonstrating specificity of the relationship between miR-338-5p differential expression with pain of a neuropathic nature. DISCUSSION/SIGNIFICANCE OF FINDINGS: These findings, along with validated targets of miR-338-5p in the NF-KB neuroinflammatory signaling pathway, suggest that miR-338-5p may serve a neuroprotective role in modulating neuroinflammation, and that its downregulation may result in maladaptive neuroplastic mechanisms contributing to the development of neuropathic pain after SCI.

The COVID-19 pandemic has exacerbated health disparities and rendered them acutely more visible. Special and underrepresented populations need to be fully integrated into the translational research process from the very beginning and all the way through. This article presents findings and rapid analysis mini-case studies from the Environmental Scan (E-Scan) of adaptive capacity and preparedness of Clinical and Translational Science Award hubs, specific to the goal of integrating special and vulnerable populations in different institutional research settings. In our discussion of the findings and case studies, we flexibly apply local adaptive capacity framework concepts and characteristics, and, whenever possible, we present ideas on how to enhance capacity in those areas, based on the challenges and practices identified through the E-Scan. Although the past year has recorded incredible achievements in vaccine development, clinical trials, diagnostics, and overall biomedical research, these successes continue to be hampered by our inability to turn them into achievements equally available and accessible to all populations.

The clinical research units (CRUs) are one of the main spaces where both translational research and science take place. However, there is a lack of information about both best practices for CRU operations and, ultimately, benchmarks to evaluate CRU performance. The Research Unit Network (RUN) was created with the purpose to enable direct communication and collaboration among CRUs. An online survey was administered to further illustrate the functionality and impact of RUN. Thirty-one individual survey responses (39.2%) were included in the final analysis. The members value RUN monthly meetings (87.1%) as the most useful aspect of this network and CRU budgeting (67.7%) and staffing (61.3%) were the most relevant topics discussed. This is followed by EPIC – Research (58.1%), delegation of authority logs, unit signatures, and policies (51.6%), COVID-19 pandemic response (41.9%), the implementation of clinical trial management system (29.0%), and protocol deviations (19.4%). The intermediate goal of RUN is to identify best practices CRUs are establishing, implementing, and sharing these experiences with the goal to adopt them in different CRUs. The network’s long-term goal is to establish standard benchmarks that can be used for evaluating the performance of CRUs across the nation.

Traditional training and funding mechanisms in academic health centers often do not support its faculty, staff, and trainees in evaluating and implementing innovative ideas, necessitating supplemental innovation programming. The University of Michigan (U-M) Frankel Cardiovascular Center partnered with U-M Fast Forward Medical Innovation (FMMI), a biomedical innovation and commercialization unit funded in part by the Clinical and Translational Science Award awarded to the Michigan Institute for Clinical & Health Research, to provide training and resources to advance ideas toward impacting patients. The program recruited faculty, trainees, staff, patients, and family members from multidisciplinary backgrounds. Engaging patients and family members expanded the ideas generated and furthered clinical relevance. Over two years, 11 project teams completed an 11-week, 16-session course on innovation and entrepreneurship concepts that incorporated workshops to progress ideas and develop a pitch for development funding. An increase in knowledge was reported in key innovation topics, such as customer discovery, assessing markets, and intellectual property. Participants reported an increase in project preparation, including obtaining stakeholder support, preparation of a development plan, readiness to apply for funding, and filing invention disclosures. This program can serve as a model for implementing training and funding mechanisms to advance innovative ideas.

OBJECTIVES/GOALS: The goal of this study is to decipher the mechanisms by which breast cancers evade the immune system to facilitate metastasis through the lymphatic vasculature. We believe this is caused, in part, by semaphorin 7A (SEMA7A) as its expression is associated with increased lymphatic vessels, tumor-associated macrophages, and metastasis in breast cancer. METHODS/STUDY POPULATION: We have observed that breast cancer cells, lymphatics, and macrophages exhibit SEMA7A-dependent expression of PD-L1 in vitro, which may make SEMA7A+ tumors more sensitive to anti-PD-1/PD-L1 immunotherapies. Therefore, we utilized multiple orthotopic, immunocompetent mouse models to determine if SEMA7A expression makes tumors more susceptible to immune checkpoint blockade. E0771 and 66cl4 mouse mammary carcinoma cells were injected orthotopically into the #4 mammary fat pads. Once tumors reached 50-100 mm^3 mice were injected intraperitoneally with 250ug of anti-PD-1, anti-PD-L1, or IgG control every third day. Tumors were measured with calipers every other day and harvested for flow cytometry to determine the effect of immune checkpoint blockade on the TME in SEMA7A+ tumors. RESULTS/ANTICIPATED RESULTS: We reveal that growth of SEMA7A overexpressing (OE) tumors, but not controls, was significantly slowed with both anti-PD-1 and anti-PD-L1 treatments. Flow cytometric analysis of cells from the TME revealed increased LECs, TAMs, and PoEMs in SEMA7A+ tumors, compared to controls – all populations had higher PD-L1 expression, which was decreased with both anti-PD-1 or anti-PD-L1. We also observed a decrease in PD-L1 expression on the tumor cells with treatment. Furthermore, LEC, TAM, and PoEM presence was decreased within the TME alongside increased presence of activated CD4 and CD8 T cells. Collectively, our results suggest that SEMA7A expression in breast cancers activates a major pathway associated with immune evasion and helps to establish a pro-tumor microenvironment. DISCUSSION/SIGNIFICANCE: SEMA7A expression establishes a pro-tumor microenvironment, which can be targeted with readily available FDA approved drugs such as immune checkpoint-based therapies. Since SEMA7A+ breast cancers have high rates of metastasis, more specific treatments for these patient populations should be explored.

OBJECTIVES/GOALS: Undergraduates experience food insecurity at rates 21% higher than the general population. Because professional students have been omitted from these studies, the goal of this project is to determine the prevalence of food insecurity among medical students at one academic institution. METHODS/STUDY POPULATION: A cross-sectional research design was used to quantify the food insecurity status of medical students at the University of Utah. The USDA’s validated 6-item Food Security Survey Module was distributed via email to all currently matriculated medical students. Student’s responses were anonymous but questions about gender and age were included. Respondents (N = 200) were scored per the module as food secure, food insecure, or very low food security. RESULTS/ANTICIPATED RESULTS: Statistical analysis included frequencies and chi-square tests. Medical students (N = 166) showed 50.6% of respondents experienced food insecurity in the past 12 months, 16.3% experienced very low food security.While there were no significant relationships between food security status and gender or age, general trends did show divorced and separated students had higher food insecurity risk 82%. A similar study in 2014 surveyed undergraduates at the same location; 51% of respondents (N = 221) experienced food insecurity. While medical students experience food insecurity at rates much higher than the national average, prevalence is lower than undergraduates at the same institution. DISCUSSION/SIGNIFICANCE OF IMPACT: Burnout and suicide in medical training are at an all-time high; professional and academic pursuits are limited when physiological needs of food security are not being met. Study results suggest, 50% of respondents are food insecure. This should inform the development of supportive programs.

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

OBJECTIVES/SPECIFIC AIMS: Reducing radiologic exams has been a focus of cost reduction in healthcare systems. The utility and justification of obtaining cross-sectional imaging (PPCSI) before surgical intervention continues to be evaluated. For peripheral artery disease (PAD) consensus guidelines regarding PPCSI do not exist and may be influenced by patient complexity, variation of disease presentation, and physician preference. The objective of this study was to determine the utility of PPCSI before percutaneous PAD intervention. METHODS/STUDY POPULATION: Patients receiving first-time endovascular revascularization procedure for PAD from 2013 to 2015 were evaluated for PPCSI done within 180 days prior to revascularization. Patient and physician demographics, perioperative characteristics, and disease distribution/severity were evaluated. The primary outcome was technical success defined as improving inflow and/or revascularization of the target outflow vessels to <50% stenosis. RESULTS/ANTICIPATED RESULTS: Of the 348 patients who underwent an attempted revascularization procedure 159 (45.7%) patients underwent PPCSI, including 151 CTA and 8 MRA. Of these, 48% were ordered by the referring provider (84% at an outside institution), and 52% were ordered by the treating physician. PPCSI was performed a median of 26 days (IQR 9-53) prior to procedure. Individual vascular surgeon practice identified PPCSI rates ranging from 31% to 70%. On multivariate analysis chronic kidney disease (OR=0.35; CI 0.17–0.73) had the strongest effect against of PPCSI, and Inpatient/ED evaluation (OR=3.20; CI 1.58–6.50), aorto-iliac (OR=2.78; CI 1.46–5.29) and femoral-popliteal occlusions (OR=2.51; CI 1.38–4.55) most strongly predicted PPCSI. After excluding 31 diagnostic procedures, technical success did not differ between endovascular procedures with PPSCI (91.3%) or without PPCSI (85.6%), p=0.11. When analyzing 89 femoral-popliteal occlusions, technical success was higher with PPCSI (88%) compared to procedures without PPSCI (69%), p=0.026. DISCUSSION/SIGNIFICANCE OF IMPACT: PPCSI use is influenced by inpatient status, chronic kidney disease, and anatomic consideration. PPCSI was not associated with overall technical success although it appeared beneficial for femoral-popliteal occlusions. Routine practices of ordering of PPCSI may not be warranted when considering technical success but may be important in treatment planning. Further studies are warranted to determine if radiation, cost, and contrast load justify PPCSI.

OBJECTIVES/GOALS: This project’s objective is to empower verbally-communicating autistic adults to express their mental health needs to mental health professionals, regardless of their training or experience in autism. By enhancing empowerment in this area, we aim to enhance their self-awareness and confidence in navigating and accessing mental health support. METHODS/STUDY POPULATION: Following the Toronto Translational Framework (TTF), our approach involves co-designing and testing an intervention directly with the autistic population through three phases. Phase I will review published literature on autism, mental health, and patient empowerment, validate our identified need through informal meetings with stakeholders, and assemble an autistic advisory committee (AAC) of 5-8 members who are able to provide informed conset without assistant, are fluent in English, and 18 years of age or older. Phase II will involve participatory design sessions with our AAC to develop a low-fidelity prototype to address the identified need. Phase III will evaluate our prototype’s effectiveness through a separate series of focus groups, which will consist of members from our target population. RESULTS/ANTICIPATED RESULTS: We are anticipating that by the end of our research, we have successfully co-designed an intervention that effectievly empowers autistic adults in their mental health journey through increasing their self-awareness and confidence in navigating and receiving mental health support. At a larger scale, results may include the empowerment of autistic adults to seek and receive mental health care from mental health professionals, regardless of professionals’ prior autism expertise. DISCUSSION/SIGNIFICANCE: By enhancing self-awareness of their mental healthcare needs and boosting self-reported confidence in communicating with mental health professionals, we aim to take the first step in creating timely, patient-centered solutions, and bridging gaps in the evolving neuro-affirmative healthcare system.

OBJECTIVES/SPECIFIC AIMS: Our study aims to create a novel state level HIV-ESRD dataset and compare patient-level characteristics on rates of transplant referral, evaluation, waitlisting, and transplantation for HIV-positive versus HIV-negative patients. Our main hypothesis is that HIV-positive patients in Georgia are less likely to be referred to kidney transplant compared with HIV-negative patients. METHODS/STUDY POPULATION: Three datasets will be merged in order to create the HIV-ESRD dataset. The datasets are United States Renal Data System (USRDS), a southeast Transplant Referral Dataset and patient-level Georgia Department of Public Health HIV Incidence Database. The resulting study population will include patients that are older than 18, but less than 70, are HIV-positive and are on dialysis in Georgia. This dataset will also identify those patients who have been referred to transplantation, have been waitlisted, and have received kidney transplants between January 2012 and December 2017. If within a 1-year period, the prevalence of HIV-positive patients referred to transplant was lower than the 1-year period prevalence of HIV-negative patients for 3 consecutive years, the dialysis facility will be classified as having a within-facility disparity. We will then characterize patient level and dialysis facility-level factors that may contribute to observed findings. Patient characteristics will include demographic, clinical data, proxies of socioeconomic status, and geospatial relationships to transplant centers and rural vs urban neighborhoods. Facility-level characteristics includes profit status (profit vs. nonprofit), total number of staff (including full-time and part-time employees), aggregate demographic and clinical facility characteristics, and total number of treated patients. RESULTS/ANTICIPATED RESULTS:. We anticipate the successful creation of the proposed dataset that will allow for accurate identification of HIV-positive patients on dialysis in Georgia.. This dataset will provide the ability to determine referral, waitlisting, and transplantation rates.. We predict the overall rate of referral, waitlisting, and kidney transplantation in HIV patients will be relatively low, and that dialysis facilities with a higher proportion of HIV-positive will have lower referral rates compared to dialysis facilities treating a higher proportion of HIV-negative patients. It is foreseen that among patient-level characteristics, the strongest predictor for decreased referral rates will be HIV serostatus and among dialysis facility factors, profit status will be associated with decreased referral rates. DISCUSSION/SIGNIFICANCE OF IMPACT: This pilot study offers the creation of the first regional dataset of HIV-ESRD patients that will include patient-level characteristic of HIV-positive patients and provide a model for other states to adopt. We will contribute improved state-level description of incidence data of HIV-positive patients on dialysis, current rates of transplant referral, waitlisting, and transplantation, and offer potential associated factors that influence these processes. This knowledge will be used to determine the next steps in improving access to care; conducting qualitative research to understand dialysis facility views on transplant in HIV patients, understanding HIV patient’s position on transplantation, providing education on the value of kidney transplant referral, and expanding the approach of combining patient level HIV data to the southeast.

ABSTRACT IMPACT: Extrapolating from mouse data we explored eosinophil content in human adipose tissue and its effect on adipocyte biology potentially leading to the discovery of novel therapeutic targets for treatment of obesity and insulin resistance. OBJECTIVES/GOALS: The interaction between immune cells and adipose tissue (AT) in obesity has not been fully elicited. Mouse models of diet-induced-obesity show AT resident eosinophils (EOS) help preserve insulin sensitivity (IS). As data in human obesity are lacking, here we explored AT-EOS content and their role in AT metabolism in subjects with and without obesity. METHODS/STUDY POPULATION: We recruited lean (L) subjects and patients with obesity (Ob) to undergo abdominal subcutaneous AT biopsy and evaluation of insulin resistance (IR) by determination of HOMA-IR. Circulating EOS were isolated from all participants under fasting conditions and exposed to high glucose (HG) or high lipids (HL) for 4 hrs. AT EOS number was assessed via FACS analysis. Circulating EOS and AT mRNA was assessed by qPCR for multiple genes involved in inflammation and cell migration. To evaluate the effect of EOS on primary human adipocytes, in vitro cultures were exposed for 4 days to either interleukin-4 (IL-4), interleukin-13 (IL-13) or to human EOS. Adipocytes mRNA levels were evaluated for genes involved in adipogenesis and lipid metabolism. RESULTS/ANTICIPATED RESULTS: 16 lean, IS subjects (BMI 22.5+ 0.4kg/m2) and 22 age-matched IR patients with obesity (BMI: 38.9 + 1.0kg/m2) participated. We observed a ratio of 2:1 in AT EOS content of L vs Ob subjects (P<0.03). To assess the reduced AT-EOS content in obesity, we evaluated expression of Chemokine-C receptor 3 (CCR3) in circulating EOS. We show decreased CCR3 mRNA levels in Ob vs L subjects (P=0.006). We expect HL in vitro experiments on peripheral EOS of L subjects to affect CCR3 mRNA levels. In AT of Ob subjects, we found a significant decreased expression of Eotaxin 2, the main EOS chemokine binding CCR3 expressed on EOS. Preliminary data from in vitro primary adipocytes culture suggest for IL-4 and IL-13 to increase mRNA level of Peroxisome Proliferator Activated Receptor Gamma (PPARG), the master regulator of adipogenesis. DISCUSSION/SIGNIFICANCE OF FINDINGS: Comparable to animal studies, we found a decrease of AT-EOS content in patients with obesity. Alterations in CCR3/Eotaxin 2 signaling may be involved. IL-4 &IL-13 are secreted predominantly by EOS and appear to directly regulate gene expression in human adipocytes. These data represent the first evidence for a novel role of EOS in human AT biology.

ABSTRACT IMPACT: Use of this novel murine model of inflammatory bowel disease (IBD) and C. difficile infection (CDI) will aid in developing new clinical approaches to predict, diagnose, and treat CDI in the IBD population. OBJECTIVES/GOALS: IBD is associated with intestinal inflammation and alterations of the gut microbiota, both of which can diminish colonization resistance to C. difficile. Here, we sought to determine if IBD is sufficient to render mice susceptible to C. difficile colonization and infection in the absence of other perturbations, such as antibiotic treatment. METHODS/STUDY POPULATION: C57BL/6 IL-10-/- mice were colonized with Helicobacter hepaticus to trigger colonic inflammation akin to human IBD. Control mice, not colonized with H. hepaticus, were pretreated with the antibiotic cefoperazone to render the gut microbiota susceptible to CDI. Mice were then gavaged with spores of the toxigenic C. difficile strain VPI 10463 and monitored for C. difficile colonization and disease. The fecal microbiota at the time of C. difficile exposure was profiled by 16S rRNA gene sequencing and analyzed using mothur. Statistical analyses were performed using R. RESULTS/ANTICIPATED RESULTS: Mice with IBD harbored significantly distinct intestinal microbial communities compared to non-IBD controls at the time of C. difficile spore exposure (14 days post-IBD trigger). Mice with IBD were susceptible to persistent C. difficile colonization, while genetically identical non-IBD controls were resistant to C. difficile colonization. Concomitant IBD and CDI was associated with significantly worse clinical and intestinal disease than unaccompanied IBD. DISCUSSION/SIGNIFICANCE OF FINDINGS: Patients with IBD who develop concurrent CDI experience increased morbidity and mortality. These studies in a novel mouse model of IBD and CDI emphasize the dual importance of host responses and alterations of the gut microbiota in susceptibility to C. difficile colonization and infection in the setting of IBD.

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

OBJECTIVES/GOALS: Asthma is a life-long, chronic lung disease that inflames and narrows the airways. Its effects on quality of life in children can be exacerbated. The goal of this study was to investigate the link between asthma, family structure and demographics and how it impacts quality of life in children. METHODS/STUDY POPULATION: We analyzed data from a cross sectional study of the 2016-2017 National Survey of Children’s Health, NSCH, to assess the relationship between determinants of health variables and the outcome variable of parent’s report of child’s perceived health status (quality of life). The study population was children under the age of 18. Data were analyzed using descriptive, bivariate analysis using Chi square, and multiple logistic regression of quality of life and family structure adjusting for confounding variables. RESULTS/ANTICIPATED RESULTS: The study included 5,687 children. Significant predictors of asthmatic children’s quality of life were severity of asthma, self-perceived mental/physical health status of adults, neighborhood safety for children (p <0.05). The interaction between family structure and asthma severity was significant indicating that asthma severity was an effect modifier. Among children with mild asthma, predictors of quality of life were self-perceived mental/physical health status of adults in the household, neighborhood safety of children, physical activity status of children (p<0.05). Among children with severe asthma, predictors were family structure and physical/mental health of adults (p<0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: This study suggests children with severe asthma who are born to single mothers with lower parental reporting of physical/mental health status had a lower quality of life. A longitudinal study could be implemented to target these three measures to improve quality of life among these children. Also, a culturally adapted intervention involving community, parents, and providers is needed to improve the quality of life of the children with asthma.

OBJECTIVES/GOALS: Insulin dosing is crucial for regulating blood glucose in cardiac surgery patients, yet it requires the use of cumbersome insulin dosing charts. To streamline this process, an electronic insulin calculator (IC) is trialed in the cardiac operating rooms and intensive care unit. This study compares glycemic control prior to and after institution of IC. METHODS/STUDY POPULATION: Using the EHR, a retrospective population of 3,164 cardiac surgery patient charts from 12/19/19 - 11/06/20 were obtained. Baseline, intra-operative, and ICU blood glucose values were obtained from each patients admission. Using this data, a baseline level of glycemic control throughout a cardiac surgery patients stay was established. A preliminary cohort of 244 patients were then chosen to be managed with the new IC. Baseline, intra-operative, and ICU blood glucose values were obtained in this group for comparison. Paired t tests were used to compare the IC group of patients to the retrospective group of patients. Additional subgroup analysis was performed to assess IC efficacy for on pump vs off pump cardiac surgeries. RESULTS/ANTICIPATED RESULTS: The 244 patients managed with the IC showed significantly reduced average blood glucose values during their time in the ICU compared to those previously not managed with the IC (185 mg% vs 153 mg%, p= 0.02). Additionally, a trend towards a reduction in last operating room blood glucose level was also noted. Lastly, average blood glucose levels were significantly reduced for patients undergoing on pump cardiac surgeries compared to off pump surgeries (157 mg% vs 149 mg%, p = 0.03). DISCUSSION/SIGNIFICANCE: Preliminary results suggest IIC to be associated with better intra and postoperative blood glucose control. More data is being collected to test its association with outcomes.

OBJECTIVES/SPECIFIC AIMS: To evaluate the determinants of non-adherence to guideline treatment and surveillance and unique barriers to care in LIU Latinos with NMIBC that will inform the development of novel patient educational materials and navigation programs that could improve adherence and thus, oncologic outcomes. METHODS/STUDY POPULATION: We will recruit 40 Latino patients with new or existing NMIBC diagnoses who present to the Urology clinic at a large, tertiary public hospital in Los Angeles from November 2018 to March 2019. Quantitative (surveys) and qualitative (semi-structured interviews) data will be collected, analyzed and integrated in order to comprehensively determine patient-level barriers to adherence. RESULTS/ANTICIPATED RESULTS: We expect to identify a unique set of patient-level barriers to adherence to NMIBC care that is unique to this population that center around 1) structural barriers to care, 2) knowledge, attitudes, and beliefs that pertain to education, acculturation, gender and values, and 3) general and disease-specific health literacy. DISCUSSION/SIGNIFICANCE OF IMPACT: The barriers to surveillance and treatment NMIBC care are significant, particularly in LIU and minority patients, which is important as non-adherence to guideline care is linked to poorer cancer outcomes. The data generated herein will inform the development of tools and programs to aid in reducing or eliminating these barriers, but also will inform discussions on the effectiveness of current clinical practices for low-income Latino patients.

OBJECTIVES/GOALS: Our objective is to investigate the antibacterial and regenerative properties of a novel AHA dental coating for the prevention and treatment of deep caries (cavities). Further, we aim to investigate and compare these properties through in vivo murine models and assessment on human saliva samples and human pulpal cells collected from clinical samples. METHODS/STUDY POPULATION: In vitro antibacterial studies were performed by collecting and culturing human salivary bacteria with AHA substrates and quantifying survival of cariogenic Sm (S. mutans). In vivo, C57BL/6 mice were treated with AHA composite fillings, infected with Sm clinical isolates, and fed a high sucrose diet with cavity formation assessment after 6 weeks. To evaluate regeneration, mice were similarly given composite with AHA or MTA (standard of care) upon pulpal exposure with regeneration quantified by microCT and histological analysis of dentin bridge formation, ALP production, and odontoblast migration. In vitro, AHA substrates were cultured with MC3T3-E1 pre-osteoblast cells and dental pulp stem cells obtained from clinical samples over 21 days, with mineralization and ALP assessed indicating osteogenesis. RESULTS/ANTICIPATED RESULTS: In vivo studies have shown the reduction of cavity formation in mice treated with AHA as well as dentin regeneration upon pulpal exposure using microCT and histological image analysis. Coinciding with these findings, AHA substrates eradicated cariogenic Sm in human saliva samples and single species cultures in vitro. Further, preliminary results in vitro have shown increased mineralization of MC3T3-E1 cells when cultured with AHA substrates in comparison to uncoated substrates. We anticipate similar increased mineralization as well as increased ALP production of human pulpal stem cells from clinical samples when cultured with AHA substrates, suggesting osteogenesis. Further, we anticipate increased odontoblast migration and ALP production upon additional analysis of in vivo tissue samples. DISCUSSION/SIGNIFICANCE: This work will elucidate the antibacterial and regenerative properties of AHA dental coatings. These results further support the translation of AHA coatings into the clinic as a novel therapeutic for the prevention and treatment of dental decay, which may overcome the limitations associated with the current treatments.