Objectives/Goals: Competencies of a principal investigator (PI) in clinical research are crucial for ensuring the success, integrity, and ethical conduct of a study. This protocol aims to assess training offerings focused on improving clinical investigator competency and build a set of best practices for training. Methods/Study Population: The authors have started by creating a committed group of key opinion leaders at Rutgers Health and across industry to advise on the process. A mixed-method assessment of the current state of clinical investigator training/education in the conduct of T1-T4 clinical trials is currently being conducted to identify existing practices. An evaluation and assessment of key competencies will be initiated. Education and training objectives and modules will subsequently be developed from this process. The program will be piloted to early career clinicians, faculty, fellows, investigators within NJACTS, and other CTSAs and later assessed for efficacy. Results/Anticipated Results: A summary and descriptive statistics of the landscape of training opportunities and the variabilities of these offerings to address the needs of these investigators is projected to exhibit the gaps in knowledge and skill set required for a PI to effectively conduct a clinical trial. This will serve as the basis for developing a micro-credential (PI Badge) by the NJACTS’ Workforce Development group. Variability across educational offerings from academic institutions, sponsor/CRO protocol/study training, and online courses may not foster applicable skills. This badge is focused on addressing a clinician’s understanding of the roles and responsibilities as they align to the Principal Investigator commitments listed in the FDA form 1572. Discussion/Significance of Impact: Whether or not micro credentialing will enhance principal investigator competency, this mixed-method assessment is poised to identify a common benchmark for success. The ability to deal with increasing protocol complexity requires investigators to be more adept at implementation and compliance to sustain their ability to conduct clinical trials.

Objectives/Goals: Community members in North Central Florida who were identified with high mental health burden, through completion of a Health Needs Assessment in our community engagement program, experienced higher rates of health disparities like food insecurity, recent marijuana use, and burden of disease compared to persons with lower mental health burden. Methods/Study Population: Data were collected through HealthStreet, a University of Florida community engagement program to reduce disparities in healthcare and research. Multivariate and logistic regression analyses were conducted to identify differences between community members with high mental health burden (HMHB; reported depression and scored 8–10 on the stress/loneliness scale) and people with low mental health burden (LMHB; no reported depression and scored 1–7 on the stress/loneliness scale) on age, sex, race, food insecurity, trust in research/researchers, recent marijuana use, high blood pressure, and anxiety. Results/Anticipated Results: Among 3,736 CMs, 21.9% (n = 819) were considered to have high mental health burden. They were statistically significantly more likely to be middle-aged (37.9% vs. 31.2% LMHB), food insecure (62.6% vs. 35.2% LMHB), self-reported marijuana use in the past 30 days (29.2% vs. 15.5% LMHB), and experienced anxiety (70.8% vs. 34.4% LMHB). There were no differences by sex or trust. Discussion/Significance of Impact: People with HMHB do in fact have more disparities than people with LMHB but trust in research(ers) is not affected. These disparities contribute to significant distress over the lifespan. HealthStreet has engaged people with higher mental health burden and provided social referrals, which is crucial to reduce disparities and improve outcomes.

Objectives/Goals: To guide UCSF in adopting a justice-based, patient-informed model of health data sharing that moves beyond traditional regulatory frameworks that focus on individual privacy and do not address public good and health equity; to create a roadmap for UCSF to implement transparent, community-engaged data governance that is both responsible and effective. Methods/Study Population: We conducted 24 hours of observation at outpatient clinics and 75 in-depth interviews with multiple stakeholder groups: patients, community advisors, and UCSF faculty and staff involved in managing and governing health and research-generated data. We used an ethnographic approach to investigate health data sharing policies and practices and explore perspectives on data governance. We were particularly interested in how data sharing is explained to patients, and what patients and community members know and think about how patient data may be used other than for clinical care. We explored faculty, staff, patient, and community member perspectives about the potential involvement of patients/communities in data governance. We also reviewed UCSF websites with information about data use and sharing policies. Results/Anticipated Results: Policy Awareness: Clear communication and improved guidelines for data use policies will enhance awareness among patients, the public, and researchers, boosting UCSF’s credibility as a data steward. Risks/Benefits: Data sharing drives scientific and clinical progress but raises concerns about privacy, profiteering, and unequal benefit distribution. Improving the Data Ecosystem: Safe data sharing and public benefit can be strengthened through centralized governance and better communication. Key growth areas include deidentification, external sharing criteria, and leakage prevention. Shared Governance: Patients and community advisors favor shared governance with public involvement, while UCSF informants were equivocal. All are concerned about equitable representation and technical training challenges. Discussion/Significance of Impact: Our recommendations to University leadership: Improve communication of data sharing policies to patients and the public. Involve patients and the public in data governance. Support investigators to ensure understanding and compliance with data use policies. Hold UCSF and collaborators accountable for transparency, equity, and public benefit.

Objectives/Goals: Evaluate use and reuse of data in biomedical fields to explore how data advances translational benefit for science and patients. Methods/Study Population: In this study, we use the metadata for 5 million data citations in the Data Citation Corpus as a source to analyze the use of datasets from biomedical fields across a number of facets, including date, affiliations, and funders. We identify examples of datasets that have been used across different research fields by analyzing the disciplinary scopes of the datasets and the publications that cite them and explore patterns of reuse of datasets beyond the original research project from which it was created. Results/Anticipated Results: The analysis will provide insights into datasets showing broad usage in biomedical fields, the patterns of data use over time, and the affiliations and funders associated with highly used datasets. The analysis will also dive into examples of datasets showing use across different fields and usage beyond the original project. The results will showcase examples of valuable datasets in translational research and serve as a basis for exploring different considerations around the impact of open datasets. Discussion/Significance of Impact: The exploration of data citations will provide insights into open data showing translational reach and an example of how the Data Citation Corpus can support evaluation of the impact of research data.

Objectives/Goals: To address the manual, time-consuming processes of validating IRB compliance and ensuring the secure delivery of i2b2 data, this project automates compliance checks, streamlines Protected Health Information (PHI) access, and provides timely, secure data availability while reducing administrative burdens and non-compliance risks. Methods/Study Population: This project enhances the i2b2 application to automate compliance processes and facilitate secure data delivery through integration with REDCap. By linking i2b2 with the IRB system, the application performs automatic compliance checks for project requests, verifying GCP and HIPAA certifications, only allowing the release of IRB-approved PHI variables, safeguarding against unauthorized data access. Manual signatures confirm non-automated compliance processes. Once verified, the application automatically creates a REDCap project, assigns user access, and securely delivers data, ensuring compliance with HIPAA regulations. Results/Anticipated Results: The automated system successfully streamlined IRB compliance checks and data delivery for i2b2 requests. Validation of certifications like GCP and HIPAA, now occurs automatically, significantly reducing the risk of non-compliance. Personnel access to data is limited to IRB-approved PHI, ensuring data security and adherence to institutional standards. The integration with REDCap has reduced manual processes, cutting data request processing time to approximately 30 minutes. Researchers and administrative staff experienced a notable decrease in administrative burden, with faster, more efficient access to approved data while maintaining full compliance with IRB and HIPAA regulations. Discussion/Significance of Impact: The lessons learned can be adapted by institutions to improve compliance efficiency and reduce administrative overhead. Implementing similar automation of certification checks and data delivery, sites can enhance data security, minimize errors, and ensure faster, compliant access to research data.

Objectives/Goals: People with insulin-treated diabetes face hypoglycemia risk due to imperfect insulin replacement and impaired counterregulation. We identified the dopamine antagonist, metoclopramide, as a potential treatment. Hypothesis: Treatment with metoclopramide will prevent the development of impaired counterregulatory response to hypoglycemia. Methods/Study Population: In a pre-clinical model, diabetes was induced in 10-week-old Sprague-Dawley rats with streptozotocin (STZ, 65 mg/kg IP). Rats were divided into three groups: 1) diabetic controls (STZ+RS, n = 6), 2) recurrent hypoglycemia (STZ+RH, n = 7), and 3) recurrent hypoglycemia + metoclopramide (STZ+RH+MET, 3 mg/kg IP, n = 7). After 3 days, all rats underwent a hyperinsulinemic (50 mU/kg/min) and hypoglycemic (~45 mg/dl) clamp. In the clinical trial, adults with Type 1 diabetes (age 20–60, ≥5 years duration) were enrolled in a phase II, double-blinded, placebo-controlled trial. Awareness status was assessed via Gold score, and subjects maintained drug regimens and underwent two hyperinsulinemic-hypoglycemic clamps (where blood glucose was lowered to 100, 65, 55, and 45 mg/dl) to assess counterregulation. Results/Anticipated Results: In the pre-clinical model, glucose infusion rates (GIR) to maintain hypoglycemia were higher in STZ+RH (27±0.9 mg/kg/min) than STZ+RS (19±0.8 mg/kg/min, p Discussion/Significance of Impact: Metoclopramide improves glucoregulatory, sympathoadrenal, and counterregulatory responses to hypoglycemia in pre-clinical models, suggesting dopaminergic regulation. While clinical data are still blinded, increased epinephrine and growth hormone responses suggest treatment may preserve or restore counterregulation.

Objectives/Goals: Bronchiolitis obliterans syndrome (BOS), a form of chronic lung allograft dysfunction (CLAD) that primarily affects the small airways, is often diagnosed too late using standard pulmonary function tests. This project aims to evaluate whether quantitative air trapping analysis can serve as an early diagnostic tool for BOS. Methods/Study Population: We performed a retrospective analysis of 134 computed tomography scans with inspiratory and expiratory protocols from 73 lung transplant recipients (48 male, 25 female). Quantitative air trapping analysis was performed by VIDA Diagnostics using a supervised machine learning technique called disease probability measure (DPM). Results/Anticipated Results: We found that lung transplant recipients exhibit significantly more air trapping compared to healthy controls and other small airway diseases, such as long COVID and cystic fibrosis. Notably, lung transplant recipients showed increased air trapping in the upper lobes. However, when separating participants into CLAD and non-CLAD groups, those meeting criteria for CLAD had significantly more air trapping in the left lower lobe. Additionally, only 2 out of 16 participants meeting CLAD criteria had less than 20% air trapping in their lungs, suggesting early involvement of the small airways. Discussion/Significance of Impact: Quantitative air trapping analysis seems to be an important diagnostic modality in the early detection of lung transplant-related small airway disease. Prospective longitudinal studies are needed to evaluate the spatial pathophysiology in these patients and to determine whether early air trapping can predict the development of CLAD.

Objectives/Goals: Despite the documented link between ultraviolet (UV) exposure and skin cancer, the potential protective role of pigmentation in preventing skin carcinogenesis is not fully understood. This project will test the hypothesis that individuals with light skin tones are more susceptible to skin cancer due to a higher accumulation of somatic mutations from UV exposure. Methods/Study Population: Skin biopsies were collected from various anatomical sites of seven donors aged 70 to 80 years with dark skin tones and used to generate clones of normal skin cells. The clones were then subjected to whole-exome and RNA sequencing. We developed a workflow that allows us to accurately detect somatic mutations in clonal expansions of individual cells with high specificity and sensitivity. For comparison, we also analyzed mutations in skin cells obtained from individuals with light skin tones, who served as our control group. In the future, we plan to employ admixture analysis to clarify the impact of race on somatic mutations by examining ancestral genetic contributions. Results/Anticipated Results: Our preliminary analysis of the existing data supports our hypothesis. The average mutation burden in individuals with light skin tone was found to be 3.778 Mut/Mb, whereas in those with dark skin tones, was only 0.818 Mut/Mb. These findings align with the prevailing hypothesis that melanin acts as a protective factor against skin cancer, as it appears to reduce the mutagenic effects of UV radiation. This suggests that individuals with darker skin may have a lower cumulative mutational load, potentially contributing to their reduced risk of developing skin cancers such as melanoma. Similarly, our results showed variations in mutational burdens across different anatomical sites, which seemed to be influenced by differing levels of UV exposure, with the highest burdens observed in areas with the greatest exposure. Discussion/Significance of Impact: Understanding how skin cancer operates in darker-skinned individuals is imperative for tailoring effective screening and targeted therapies to meet their specific needs. In the long run, elucidating the mechanisms underlying skin cancer development in this demographic will help refine screening protocols and prevention recommendations.

Objectives/Goals: To reduce cervical cancer in the USA, it is essential to identify the reasons underlying the low HPV vaccination rates. This study aims to identify knowledge and beliefs about HPV vaccination among women in rural areas. The knowledge gained from this study will directly lead to developing an education tool tailored specifically for women in rural areas. Methods/Study Population: We conducted a cross-sectional study from November 2022 to July 2023. We recruited women (n  =  141) who visited a mobile health unit in rural North Louisiana. The inclusion criteria were women aged 25–64 years who spoke English, had not undergone a total hysterectomy, and had no history of cervical cancer. Data collected included sociodemographic characteristics, sexual history, awareness and knowledge of HPV infection, cervical cancer, genital warts, and HPV vaccination, the perceived risk of acquiring genital HPV infection and developing cervical cancer, and the willingness to receive an HPV vaccine. Descriptive statistics were used to evaluate participant responses. Written informed consent was obtained before completing the self-administered questionnaire. Results/Anticipated Results: Our findings showed significant gaps in HPV vaccine knowledge and uptake. Approximately 40% of the participants were unaware of the HPV vaccine, 96.5% had never received it, and 91.4% had never been offered it. However, 42% indicated a willingness to consider vaccination if it were offered. Factors influencing their decision to vaccinate against HPV included a family history of cervical cancer (44.7%), having multiple sexual partners (48%), and engaging in unprotected sex (61.7%). Furthermore, there was significant uncertainty surrounding the vaccine, with 65.2% of participants unsure about its safety, 66% unaware of potential health risks, and 47.5% uncertain about its effectiveness in preventing HPV. Discussion/Significance of Impact: Our research emphasizes the need for tailored behavioral interventions to address knowledge gaps about the HPV vaccine. Low HPV vaccination rates in rural areas contribute to health disparities in cervical cancer. Implementing educational interventions in healthcare settings can enhance vaccination rates and mitigate the risk of cervical cancer.

Objectives/Goals: Obesity has been classified as a global epidemic and origin of numerous health issues. The central hypothesis of this study is that psychological measures, DNA methylation, and gene transcription will predict obesity-related outcomes after lifestyle interventions, and such interventions may alter DNA methylation profiles. Methods/Study Population: This study consisted of a randomized-controlled trial examining the effects of lifestyle +/- stress reduction interventions in 285 highly stressed parents with obesity, followed for 2 years. Full participants received nutrition and activity counseling, and were randomized to either a stress reduction intervention or a contact control. Those who otherwise qualified for the study but unable to fully participate were included in a no intervention control group. The intervention consisted of 12 weeks of nutrition and activity counseling +/- 2-hour weekly stress reduction interventions using MBSR and CBT-based strategies. DNA methylation was assessed using Illumina EPIC arrays. Results/Anticipated Results: Using linear mixed models (LMMs), this study will first examine the hypothesis that baseline psychological measures and pre-existing methylation sites associated with obesity and glycemic control (e.g., ABCG1, ATP10A, TXNIP, SREBF1, RNF39, and SOCS3) predict changes in BMI, HOMA-IR, and HgbA1C post-intervention and at 1 and 2 year follow-ups. Using sites that demonstrate statistical significance, we will develop a polymethylation risk score predictor of change in BMI. Next, we will examine the hypothesis that interventions which reduce obesity may also lead to improvements in epigenetic aging using LMMs to determine if changes in BMI or HOMA-IR predict changes in epigenetic age acceleration over the course of the study. Discussion/Significance of Impact: This work examines whether psychological factors and/or epigenetic markers may be used in patient stratification at initiation of treatment, enabling improved treatment selection, fewer years of obesity and decreased risk of comorbidities. This proposal also asks whether lifestyle interventions impact the aging process itself.

Objectives/Goals: In mice, it has been shown that loss of Cib2 (calcium and integrin-binding protein 2) results in progressive retinal disease that recapitulates many characteristics of age-related macular degeneration (AMD). This study aims to characterize transcriptional changes in the retinal pigment epithelium (RPE) that underlie this disease process. Methods/Study Population: RPE tissue samples, pooled from 2–3 mice for each biological group, were collected from Cib2-KO and wildtype (WT) mice at two (young) and eight (aged) months of age. Bulk mRNA sequencing was performed using the Illumina HiSeq 4000. Reads were aligned to the UCSC mouse reference genome and quantified using HTSeq. Significant differentially expressed genes (DEGs) between mouse genotype and age groups were assessed using DESeq. CLICK unsupervised clustering followed by gene ontology analysis was performed to identify cellular processes and molecular pathways affected by loss of Cib2 as well as age. Results/Anticipated Results: CLICK analysis revealed several functional pathways that are differentially expressed between sample groups. For example, in both young and aged mice, pathways upregulated in Cib2-KO samples included calcium signaling, RhoA signaling, and integrin signaling. Uniquely downregulated DEGs in young Cib2-KO animals were related to complement and coagulation cascades, LXR/RXR activation (related to lipid synthesis and transport), and phagosomes. Aged Cib2-KO mice displayed the most significant downregulation of genes in the phototransduction pathway, indicating temporal changes in functional pathways that correlate with disease progression. Next steps in analysis include investigating patterns in RPE- and AMD-signature gene sets that may identify molecular pathways more specific to human disease. Discussion/Significance of Impact: Many current studies investigate the role of complement activation, vesicle trafficking, and ion transport as top contributors to AMD development. We identified DEGs paralleling many of these molecular pathways in Cib2-KO mice, highlighting their potential as a model to study age-related RPE pathologies and evaluate therapeutic interventions.

Objectives/Goals: Monensin is FDA approved for use in veterinary medicine. Recent studies pointed to its potent anticancer activity. Since de novo drug discovery process typically takes 10 to 15 years and requires an investment of approximately $1.3 to $3 billion, drug repositioning can bypass several steps in this process and increase the potential for success. Methods/Study Population: Cell viability assays were conducted on human MDA-MB-231, MDA-MB-468, and MCF10A breast cancer cell lines and mouse EO771 and 4T1 breast cancer cell lines. MDA-MB-231 cell line was used in all the studies unless specified otherwise. Time course levels of Bcl-2, Bak, p62, and LC3II were assessed via Western blotting with GAPDH as a loading control. Proteomics analysis was conducted by the IDEA National Resource for Quantitative Proteomics. Time course levels of major histocompatibility complex (MHC) I and II and calreticulin were evaluated using flow cytometry. At least three biological replicates have been conducted for each experiment. Results/Anticipated Results: Monensin and several of its novel analogs were potent toward human and mouse breast cancer cell lines. Furthermore, they induced apoptotic cell death as evidenced by Annexin V/PI assay, downregulation of Bcl-2, and upregulation of Bak in MDA-MB-231 cells. Proteomics analysis revealed that several molecular pathways related to MHC class I and II antigen presentation were significantly altered following treatment with these compounds. Additionally, monensin and its analogs significantly increased the expression of MHC class I and II. Our studies also showed that monensin and its analogs increase the surface calreticulin levels. Treatment of MDA-MB-231 cells with these compounds also resulted in an increase in p62 and LC3II expression, suggesting a disruption of the autophagic process. Discussion/Significance of Impact: These results suggest that monensin and its analogs not only exhibit anti-breast cancer cell activity but also modulate immune-related pathways. By disrupting autophagy and enhancing calreticulin levels, these compounds may potentiate antitumor immune responses, providing a promising avenue for drug repositioning in cancer therapy.

Objectives/Goals: Lack of comparative data limits research operations quality improvement (QI). The Northwest Participant and Clinical Interactions (NW PCI) Network, a group of 17 unaffiliated university and health system-based research centers, built an operations dashboard to track efficiency and enable multisite QI projects. Methods/Study Population: A Data Governance Working Group was assembled to establish shared data governance, draft nondisclosure (NDA), and data transfer and use agreements (DTUA) suitable across organizations and standardize research operations metric definitions. Sites in the NW PCI Network were recruited to participate in a pilot program to assess data sharing and governance infrastructure, data collection, upload procedures, and data visualization tools. The NW PCI Coordinating Center developed an analytical data dashboard of research operations metrics and conducted semi-structured interviews with participating sites to understand barriers and facilitators of program success. Results/Anticipated Results: Four sites (2 health systems, 2 universities) were recruited for the pilot and reviewed and executed NDAs and DTUAs. Three of the sites have submitted data for a total of 1,405 studies. Of the 24 requested data operations metrics (e.g., study startup, recruitment, implementation, and basic study information), 71% of the metrics were submitted by all three sites (n = 17), 25% were submitted by at least one site (n = 6), and 4% were not submitted by any site (n = 1). Interviews with sites after data submission found areas for improvements (clarification of data definitions, efficiency of data upload process) and positive effects for sites (e.g., process improved insight into own data operations). Discussion/Significance of Impact: Unaffiliated research centers created data governance procedures to enable sharing of operations data. Pilot sites successfully loaded most but not all operations data to the dashboard. Interviews identified process limitations and opportunities for improvement to inform expansion to all NW PCI sites.

Objectives/Goals: Understand the impact of sarcopenia on the main respiratory muscle, the diaphragm (DIAm). We hypothesize that in the DIAm of older (i.e., 24 months) compared to younger (i.e., 6 months) rats, maximum specific force (P0) is reduced, maximum shortening velocity (Vmax) is slower, maximum power output is reduced, and endurance is improved. Methods/Study Population: Mid-costal DIAm strips were excised from 6-month (n = 8; 4 female and 4 male) and 24-month (n = 8; 4 female and 4 male) rats. The DIAm was stimulated using platinum plate electrodes, and mechanical and endurance properties were measured (at 26oC). Results/Anticipated Results: In the DIAm, maximum tetanic formce (P0) decreased by ~35%, maximum velocity of shortening (Vmax) slowed by ~20%, and peak power output was reduced by ~35% in 24-month compared to 6-month rats. During repetitive isovelocity (30% Vmax; approximating peak power output) contractions, endurance (the period during which power output was sustained) of the DIAm was unaffected by aging. Corresponding with previous findings, Discussion/Significance of Impact: The changes in DIAm mechanical performance corresponded to an age-related atrophy of type IIx/IIb muscle fibers. We conclude that force generation and endurance of the DIAm required for breathing motor function is preserved in old age, while DIAm sarcopenia does impair more forceful expulsive airway clearance and voiding behaviors.

Objectives/Goals: Skilled clinical research professionals are essential to efficient and effective research teams, but many undergraduate students are not aware of staff-level careers in the field. To address this, the Michigan Institute for Clinical and Health Research launched the Clinical and Health Research Professional (CHRP) Pathways Program. Methods/Study Population: The 10-week hybrid CHRP Pathways Program was piloted in Summer 2024. Undergraduate students interested in research careers were recruited from local universities and community colleges. The program consisted of 8 hours/week of didactic curriculum guided by ECRPTQ competencies and 32 hours/week of mentored work experience on research teams. Classroom activities were completed with participants from a partner program within the university. Surveys were administered at 3 weeks and 10 weeks along with an end-of-program focus group to assess acceptability and self-reported learning outcomes. Results/Anticipated Results: The pilot CHRP Pathways Program enrolled 3 students and was well-received by participating students and mentor research teams. Students agreed that they would recommend the program to their classmates. Students indicated that the program helped them understand the role of a study coordinator, provided insight into research career paths, and helped them form professional relationships. Self-reported confidence levels in a range of research competencies increased. Two students who completed the 10-week program chose to continue working part-time with their research teams while continuing their undergraduate studies. Discussion/Significance of Impact: Experiences like the CHRP Pathways Program provide valuable exposure to staff-level research career opportunities for students engaged in health science studies. They can address an existing workforce gap by equipping college graduates with relevant work experience and basic research competencies.

Objectives/Goals: Obtaining reliable clinical research professional (CRP) employment data within and across Clinical and Translational Science Awards (CTSA) institutions is an ongoing challenge. We describe an intra-institutional approach implemented to generate routine and accurate CRP data reports to monitor and evaluate CRP career progression and assist in formation of an institutional CRP network. Methods/Study Population: A research job family with 47 job series including human, animal, and laboratory research positions was implemented at Virginia Commonwealth University (VCU). However, CRP job satisfaction surveys and evaluations could not be confidently interpreted due to the confounding animal and laboratory research positions. Led by VCU Clinical and Translational Science Awards Workforce Development a cross-functional team was formed to isolate specific CRP positions. The team included CRP front-line staff and managers partnering with VCU Human Resource Information Systems. Identified were 39 unique CRP positions across 13 distinct job series. This identification provides CRP new hire and job specific data for evaluation and tracking as well as the ability for CRP directed communications. Results/Anticipated Results: Initial and monthly HR data reports were used to develop an institutional CRP list-serv for 325–350 allowing for targeted CRP communications within a decentralized environment. Bimonthly HR data reports identify university new hires and internal transfers into any of the 39 unique jobs within 0 – 12 days of hire. Twelve unique data points are provided (name, email, current position hire date, job code, job title, working title, department, division, supervisor’s name, job title, email, and job code) allowing for tracking and analysis of retention rates, career progression, and lateral movement among other outcomes. Collaboration led by VCU Clinical and Translational Science Awards Workforce Development team provides the representative CRP staff, managers, and institutional leadership with a renewed confidence interpreting CRP employment data. Discussion/Significance of Impact: The team science approach to identify and develop routine and real-time reporting of CRP job specific data provides a rich source of information. The information is used to evaluate CRP job satisfaction and factors contributing to CRP retention, engage in future mixed-methods research, and support the formation of an institutional CRP network.

Objectives/Goals: The goals of this research are to 1) determine the prevalence of perinatal doula services use in Virginia, with a focus on individuals with substance use disorders (SUD), 2) evaluate awareness of doulas among pregnant and postpartum people with SUD, and 3) assess provider knowledge and interaction with doulas for the care of this population. Methods/Study Population: Both quantitative and qualitative methods will be used to evaluate patient and healthcare provider knowledge regarding doula services and the patient–doula–healthcare provider relationship. Surveys and semi-structured interviews will be administered to doulas, pregnant and postpartum women, and healthcare providers in this mixed-methods approach. Information from the Centers for Medicare and Medicaid Services National Provider Identifier (NPI) Registry, and doula training programs will be utilized to recruit doulas for participation. Paper and online recruitment materials will be posted to engage pregnant and postpartum individuals. Healthcare provider recruitment will occur via the NPI Registry along with contacting physicians’ practices. SAS 9.4 and NVivo will be utilized for analysis. Results/Anticipated Results: This proposed research will be an initial assessment of the current state of doula services utilization, mothers’ knowledge of doulas and their purpose, and healthcare providers’ awareness of and partnership with doulas to provide optimal birthing and postpartum experiences to the pregnant and parenting population with and without SUD. Results from this study will be disseminated to community doulas, pregnant people and mothers with substance use disorders, and relevant healthcare providers to decrease barriers to doula care and advocate for consistent, systematic documentation of doula services in the medical record and in public health surveillance systems. Discussion/Significance of Impact: This study will be the first study to assess doula services utilization in Virginia, with a specific focus on pregnant and postpartum women with substance use disorders. This work will support advocacy for improved data capture and utilization regarding doula services in order to reduce barriers to care and improve perinatal outcomes.

Objectives/Goals: Methicillin-resistant Staphylococcus aureus (MRSA) is a human bacterial pathogen and is classified as a serious threat. MRSA has become resistant to most B-lactam antibiotics (penicillins and cephalosporins). The goal of this study is to identify an antibiotic adjuvant capable of resurrecting B-lactams for the treatment of MRSA infections. Methods/Study Population: A fluorescence-reporter assay was used to screen a compound library. Minimum-inhibitory concentrations were assessed for the compounds against various MSSA and MRSA strains. A common resistance mechanism to B-lactams by MRSA is by the function of the bla operon. One gene in this operon encodes for a B-lactam sensor/signal transducer protein BlaR, the primary target of this study. Inhibition of BlaR by compound 1 (best potentiator of oxacillin) was studied by nano-differential scanning fluorimetry (nanoDSF), surface plasmon resonance (SPR), scanning electron microscopy (SEM), and time-kill assays. Results/Anticipated Results: We identified 80 compound hits from a 1,974-compound NCI library. Twenty-four compounds showed potentiating ability (2- to 4,096-fold decrease in MIC for oxacillin). Seven compounds exhibited melting temperature shifts by nanoDSF of BlaR, indicating binding. SPR determined compound 1 has a binding affinity of 31 micromolar to BlaR-SD. SEM images showed disruption in the S. aureus cell wall on exposure to compound 1 and oxacillin. S. aureus N315 showed 3-log reduction in bacterial count treated with a mixture of compound 1 and oxacillin. Discussion/Significance of Impact: Compound 1 targets BlaR-SD, which restores S. aureus susceptibility to treatment by oxacillin. There are currently few antibiotics available in the clinic capable of treating MRSA infections. The combination hold promise of a treatment option for MRSA.

Objectives/Goals: The study focuses on developing a wound patch that employs a biocompatible matrix which incorporates mesenchymal stem cells (MSCs) with wound healing and antimicrobial properties, along with antimicrobial metallic nanoparticles covered with keratinocytes derived from induced pluripotent stem cells to replicate the skin’s barrier function. Methods/Study Population: In vitro experiments will be conducted to combine bacteria with MSCs and metallic nanoparticles to assess whether bacterial killing is improved by this combination. The MSCs will then be evaluated in the presence of the nanoparticles to confirm that their functionality and phenotype are not altered. To verify the cells’ functional integrity, they will undergo trilineage differentiation, surface marker phenotypic testing, and evaluation of their capacity to inhibit lymphocyte proliferation in the presence of the nanoparticles. Subsequently, this living bandage will be created using a biomatrix embedded with induced pluripotent stem cell-derived keratinocytes and tested on a canine wound model to study the impact on healing. The model will assess the rate of healing and cellular response at weekly intervals until healed. Results/Anticipated Results: The combination of mesenchymal stem cells and antimicrobial nanoparticles works synergistically to enhance bacterial killing in vitro with S. aureus. The presence of the nanoparticles in combination with MSC did not affect the ability of the MSC to undergo trilineage differentiation. We anticipate that the surface phenotype will be similarly unaffected. In addition, we expect that the presence of the nanoparticles should not interfere with the ability of MSC to suppress lymphocyte proliferation. Utilization of the wound patch in the in vivo canine wound model is expected to enhance healing and prevent infection. We expect that we will observe a shift in the cellular composition of the wound with less inflammatory cells and more M2 or wound healing anti-inflammatory monocytes. Discussion/Significance of Impact: The incidence of resistant infections with no pharmacologic therapy available are on the rise. The development of an antimicrobial living bandage that increases the body’s ability to fight off infection, while providing a barrier to reinfection would provide a new way to treat infections regardless of their acquired antibacterial resistance.

Objectives/Goals: Cutaneous lupus erythematosus (CLE) is an inflammatory skin manifestation of lupus. CLE lesions are frequently colonized by Staphylococcus aureus, a microbe known to promote IFN production and inflammation. Here, we investigate whether type I IFN and inflammatory gene signatures in CLE lesions can be modulated with a topical antibiotic treatment. Methods/Study Population: SLE patients with active CLE lesions (n = 12) were recruited and randomized into a week of topical treatment with either 2% mupirocin or petroleum jelly vehicle. Paired samples were collected before and after 7 days of treatment to assess microbial lesional skin responses. Microbial samples from nares and lesional skin were used to determine baseline and posttreatment Staphylococcus abundance and microbial community profiles by 16S rRNA gene sequencing. Inflammatory responses were evaluated by bulk RNA sequencing of lesional skin biopsies. Immunophenotyping of CLE lesions was performed using CIBERSORTx to deconvolute the RNA-seq data into predicted cell populations impacted by treatment. Results/Anticipated Results: We identified 173 differentially expressed genes in CLE lesions after topical mupirocin treatment. Mupirocin treatment decreased the abundance of Staphylococcus associated with CLE lesions without altering the overall diversity of the skin microbiota relative to vehicle. Decreased lesional Staphylococcus burden correlated with decreased IFN pathway signaling and inflammatory gene expression and increased barrier dysfunction. Interestingly, mupirocin treatment lowered skin monocyte levels, and this mupirocin-associated depletion of monocytes correlated with decreased inflammatory gene expression. Discussion/Significance of Impact: Mupirocin treatment decreased lesional Staphylococcus burden and this correlated with decreased IFN signaling and inflammatory gene expression. This study suggests a topical antibiotic could be employed to decrease lupus skin inflammation and type I IFN responses by reducing Staphylococcus colonization.