The fifth in a 5-part series on the clinical and translational sciences educational pipeline, this paper focuses on strategies for developing leadership capacity among senior faculty and administrators responsible for clinical and translational science (CTS) research. Although progression in academic rank recognizes scientific excellence in research or scholarship, neither disciplinary training nor experience alone prepare senior faculty for the leadership challenges they inevitably face. Yet these faculty are increasingly responsible for multidisciplinary teams working within complex organizations with unclear or conflicting incentives that demand innovation. In academic health centers with Clinical and Translational Science Awards (CTSAs), investing in leadership often includes career development support in the CTSA education and training pillar programs. Only a few CTSAs have taken an intentional approach to developing senior leadership capacity, however, and still fewer have focused specifically on building such capacity for current CTS leaders within the context of a growing emphasis on team science. This manuscript explains the need for senior leadership training and describes an established example of such a program, the year-long Leadership for Innovative Team Science program for senior CTS researchers at the University of Colorado. The development of the program over time, topical elements, and participant perspectives are provided.

Interim analyses in clinical trials can take on a multitude of forms. They are often used to guide Data and Safety Monitoring Board (DSMB) recommendations to study teams regarding recruitment targets for large, later-phase clinical trials. As collaborative biostatisticians working and teaching in multiple fields of research and across a broad array of trial phases, we note the large heterogeneity and confusion surrounding interim analyses in clinical trials. Thus, in this paper, we aim to provide a general overview and guidance on interim analyses for a nonstatistical audience. We explain each of the following types of interim analyses: efficacy, futility, safety, and sample size re-estimation, and we provide the reader with reasoning, examples, and implications for each. We emphasize that while the types of interim analyses employed may differ depending on the nature of the study, we would always recommend prespecification of the interim analytic plan to the extent possible with risk mitigation and trial integrity remaining a priority. Finally, we posit that interim analyses should be used as tools to help the DSMB make informed decisions in the context of the overarching study. They should generally not be deemed binding, and they should not be reviewed in isolation.

The US biomedical research workforce suffers from systemic barriers causing insufficient diversity and perpetuating inequity. To inform programming enhancing graduate program access, we implemented a formative mixed-method study to identify needed supports for program applications and graduate program success. Overall, results indicate value in added supports for understanding application needs, network development, critical thinking, time management, and reading academic/scientific literature. We find selected differences for underrepresented minority (URM) students compared to others, including in the value of psychosocial supports. This work can inform broader efforts to enhance graduate school access and provides foundation for further understanding of URM students’ experiences.

Community Engagement and Outreach (CEO) Cores are a key component of the IDeA Clinical and Translational Research (IDeA-CTR) Program, serving to develop and implement capacity-building initiatives that empower communities to engage and collaborate with investigators in community-engaged research. To strengthen collaboration and shared learning across sites, a CEO Special Interest Group was established. In 2022, the group conducted a landscape analysis to systematically characterize and categorize CEO Core activities across the IDeA-CTR network, with a focus on capacity-building efforts related to knowledge transfer, co-learning, and infrastructure development. The analysis revealed several common approaches to community engagement and capacity building across sites, as well as distinctive strategies tailored to the specific needs and communities served. Multiple innovative practices were identified and disseminated across the network, highlighting opportunities to enhance collective impact and inform future community-engaged efforts.

ABSTRACT IMPACT: This project demonstrates that addressing low-value care, which has the potential to cause patient harm, relies on novel data tools and collaboration between health system and research stakeholders. OBJECTIVES/GOALS: Reducing low-value care, or patient care that offers no net benefit in specific clinical scenarios, is an important approach to improving value as it can simultaneously lower health care spending and improve quality. We describe an initiative to identify such care in a large statewide employer. METHODS/STUDY POPULATION: Claims data for self-funded University of California (UC) Preferred Provider Organization (PPO) plan members during 2019 were abstracted from the University of California Health (UCH) Clinical Data Warehouse, a unique central database that includes electronic medical record data from >5 million patients across UC medical campuses and all claims from UC self-funded health plans. UCH spans six academic health systems across California. The Milliman MedInsight Health Waste Calculator, a proprietary algorithm-based software tool, was used to identify low-value care and estimate associated spending. The HWC measures 48 low-value services using recommendations from the Choosing Wisely Campaign, the US Preventive Services Task Force, and other clinical specialty guidelines. RESULTS/ANTICIPATED RESULTS: Of 43,882 members of the UC PPO, 11,174 (25.4%) received at least one low-value service. The HWC identified 50,103 eligible services and classified 35% as low-value. Total spending on low-value services ranged between $2,209,516 and $5,089,866, based on a more or less conservative estimate. Across the five sites, the proportion of low-value services ranged from 31% to 39%. Five services comprised 65% of costs from low-value care: annual EKGs, preoperative baseline labs for low-risk surgeries, vitamin D deficiency screening, imaging for eye disease, and headache imaging. The top five services by order frequency were annual EKGs, vitamin D tests, preoperative labs, antibiotics for upper respiratory infections, and imaging for eye disease. DISCUSSION/SIGNIFICANCE OF FINDINGS: Low-value care is prevalent and costly within a large statewide employer. Collaborative multidisciplinary partnerships between employers, health systems, informatics, and researchers can leverage existing data to identify opportunities for improving the value of care for covered populations.

OBJECTIVES/GOALS: We prioritize Chlamydia’s public health impact, aim to develop rVCG-MECA for practical use, study robust immunity for effective strategies, and assess animal models for human vaccination adaptation. Our work highlights rVCG-MECA’s translational significance in public health. METHODS/STUDY POPULATION: Female Mice C57BL/6J mice (N=8) were immunized intramuscularly(IM) and intranasally(IN) and boosted twice, two weeks apart, with rVCG-MECA, once with live Chlamydia (C. trachomatis serovar D elementary bodies) and PBS. Specific mucosal and systemic immune responses were characterized. Vaccine efficacy was determined from chlamydia shedding following the transcervical challenge. Additionally, Chlamydia-specific cytokine (IFN-γ and IL-4) production by splenic and ILN T cells was assessed after 16 weeks RESULTS/ANTICIPATED RESULTS: Immunization with rVCG-MECA via intramuscular and intranasal routes triggered notable humoral responses in systemic and mucosal tissues. Intramuscular vaccination produced higher IgG2c levels in both tissues, while intranasal vaccination led to elevated IgA levels in mucosal tissues. rVCG-MECA-immunized mice exhibited significantly higher IFN-γ (Th1) secretion compared to IL-4 (Th2), with intramuscular immunization showing the highest IFN-γ levels. These findings anticipate robust immune responses, promising protection against Chlamydia, particularly through the intra muscular route. Overall, our results support rVCG-MECA as a promising Chlamydia vaccine, aligned with public health goals. DISCUSSION/SIGNIFICANCE: This study suggests that IM and IN immunization with rVCG-MECA induces immune effectors such as IFN-gamma and IgG2c that mediate chlamydial clearance in the genetical tract.

Objectives/Goals: Nutrition status is an overlooked, modifiable driver of total joint arthroplasty (TJA) recovery that can reduce complication rates. Therefore, we will profile preoperative nutrition status in TJA patients using integrated clinical measures, identify metrics that predict postoperative complications, and validate their clinical utility. Methods/Study Population: We will enroll 800 elective hip (THA), knee (TKA), and shoulder (TSA) arthroplasty patients. Assessments will include blood biomarkers (albumin, lymphocytes, vitamin D/A, zinc, HbA1c, lipids), bioelectrical impedance (BIA) for body composition metrics (phase angle, body fat%, skeletal muscle index: SMI, visceral adipose area: VFA), and handgrip strength (muscle quality index: MQI). Biomarkers will inform on deficiencies, immunity, and metabolic health, reflecting surgical readiness. We will compute Onodera’s Prognostic Nutritional Index (OPNI), SMI, and MQI and will evaluate infections, hospitalizations, revisions, and prolonged stay via records and encounters at 3 months and 1 year postop. Analyses will link nutrition metrics to complications to identify the strongest, actionable predictors. Results/Anticipated Results: Among the first 100 arthroplasty participants assessed (THA: 43%, TKA: 49%, TSA: 8%), 56% were male (age 66.3±10.4 yrs). Mean lymphocytes was 1.9±0.6×109/L (normal: 1.0-4.8×109/L). Zinc averaged 59.9±11.6 µg/dL (normal: 60-120 µg/dL) with 47% deficient (50.9±6.6 µg/dL) versus 53% in normal range (66.8±5.0 µg/dL). Average vitamin D was 36.3 ng/mL (sufficient: >30 ng/mL), with 42% of patients low (20.8±5.6 ng/mL) versus 48% sufficient (47.8±15.1 ng/mL). Body composition measures: BMI 30.2±5.9 kg/m2 (2% underweight, 18% normal, 27% overweight, 27% obese 1, 14% obese 2, 6% obese 3); body fat 35.1±9.4%, VFA 155.8±55.3 cm2 (healthy: <100 cm2), SMI 7.9±1.5 kg/m2, phase angle 5.0±0.8°; and MQI 1.4±0.4. Comorbidities: T2DM 18%, HTN 67%, hypercholesterolemia 28%, CVD 37%; and 8% used GLP-1 medications. Discussion/Significance of Impact: These early data show high zinc and vitamin D deficiency (47% and 42%, respectively), obesity (47% class I–III), and substantial comorbidities, underscoring the need for standardized nutrition screening in orthopedics. These findings justify targeted preop interventions and support an NIH R01 focused on reducing complications.

Objectives/Goals: Adolescence is a critical period where brain networks are thought to be influenced by environmental factors. This presentation examines violence exposure’s impact on brain connectivity and identifies potential protective factors. Methods/Study Population: A secondary data analysis was conducted using data from a subsample of the Adolescent Brain Cognitive Development Study (release 5.1). Youth who completed victimization questionnaires at two time points were eligible for inclusion, resulting in 2016 participants. Linear regression was utilized to analyze associations between violence exposure measured by the juvenile victimization questionnaire and functional connectivity of specified regions of interests using the Gordon functional parcellation for cortical regions and the Freesurfer parcellation for subcortical regions. Moderation analysis will be utilized to assess the effects of peer and caregiver support on the associations between violence exposure and functional connectivity, currently ongoing. Results/Anticipated Results: Between 18 and 59% of the sample reported experiencing at least one form of violence exposure, with racial differences noted in missing versus complete data. Multiple domains of violence and cumulative exposure were associated with both increased and decreased functional connectivity across within-network, between-network, and network-subcortical regions. At baseline, internet violence was linked to lower within-network connectivity, while peer victimization was associated with higher connectivity at both baseline and follow-up. Between network analysis showed lower connectivity with witnessing violence at baseline and higher connectivity with internet victimization at follow-up. Discussion/Significance of Impact: These findings emphasize the need for further exploration of the underlying mechanisms that link violence exposure to developmental trajectories and identification of protective factors such as caregiver and peer support, to inform interventions and promote resilience in affected youth.

Objectives/Goals: In psychosis spectrum disorders, diffusion tensor imaging studies consistently report reduced fractional anisotropy in the superior longitudinal fasciculus, a major tract linking frontal and parietal cortices, yet the neurochemical underpinnings of these microstructural abnormalities remain poorly understood. Methods/Study Population: We collected diffusion MRI scans and single-voxel PRESS magnetic resonance spectroscopy scans both in parietal and frontal white matter in 48 medication naïve patients and 39 healthy controls. Fractional anisotropy within the superior longitudinal fasciculus was quantified in using TORTOISE and compared with frontal and parietal neurometabolic profiles processed with LC model and partial volume corrected. Results/Anticipated Results: Myoinositol and TNAA were significantly different between patients and controls in both frontal and parietal white matter. Fractional anisotropy of the superior longitudinal fasciculus was significantly different between patients and controls. Both myoinositol and TNAA were associated with superior longitudinal fasciculus fractional anisotropy when modeled independently, but when jointly modeled only TNAA was significantly associated with fractional anisotropy in the parietal white matter while only myoinositol was significant in the frontal white matter. Both parietal and frontal white matter had a significantly greater association between myoinositol and fractional anisotropy than controls. Discussion/Significance of Impact: Disruptions in neuronal integrity and glial dysfunction likely contribute to decreased fractional anisotropy in frontoparietal white matter in first-episode psychosis spectrum disorders. These disturbances are not related to medication effects.

OBJECTIVES/GOALS: Artificial intelligence (AI) depends on quality machine learning (ML) algorithms constructed with high-quality training data. This TL1 trainee project develops a disease-agnostic computable phenotype framework for ML algorithm construction, modeling male puberty as a case example. METHODS/STUDY POPULATION: A computable phenotype of male puberty was constructed to answer the question: “Does early pubertal timing increase the risk of developing type II diabetes (T2D) in males?” A computable phenotype of males < 18 years old was created in the TriNetX© Diamond Network utilizing Boolean operator data queries. TriNetX© contains patient electronic health record information (ICD-10 diagnoses, anthropometric measures). An exploratory analysis of patient counts reflecting various computable phenotypes allowed for outcome (T2D) comparison of males diagnosed with precocious puberty (E30.1, ICD code for early pubertal timing) to those without, controlling for body mass index (BMI). RESULTS/ANTICIPATED RESULTS: Subjects (n=12,996,132) displayed the following computable phenotype: Male, < 18 years old, without ever having a BMI documented >85th percentile. Males diagnosed with precocious puberty (E30.1) were 6.89 times more likely to develop T2D when aged 14-18 years old than those without (OR 6.89, 95% CI: 5.17-9.19, p<0.0001). Next steps involve training a ML model on each computable phenotype groupings’ health data, with anticipated results identifying underlying salient pathophysiologic variables. A generalized computable phenotype approach is further developed to: 1) explore clinical questions in large databases like TriNetX©, and 2) model disease development with AI/ML algorithm construction. DISCUSSION/SIGNIFICANCE: Computed phenotypes reveal males with precocious puberty may have increased T2D risk. Next steps utilize subject data to train an AI/ML algorithm, model development to identify salient pathophysiologic variables, and synthesize a generalized AI/ML developmental research framework for dissemination.

OBJECTIVES/GOALS: Research on the safety of perinatal kratom use - an herb that acts on opioid receptors - is scarce. Our transdisciplinary clinical and translational science (CTS) team is conducting parallel qualitative analyses of subreddit posts related to kratom use during (1) pregnancy and (2) breastfeeding. METHODS/STUDY POPULATION: Pregnancy- and breastfeeding-related keywords are being used to extract posts and selected metadata from the following subreddit communities: r/kratom, r/quittingkratom, r/pregnant, and/or r/breastfeeding. After the removal of duplicate posts, posts written in a non-English language and those that state in the post text and/or title that they were published by minors ( RESULTS/ANTICIPATED RESULTS: Among the eligible posts, the number of unique usernames of the sources publishing the posts; the range of publication dates; and the mean, median, & range of the number of comments per post will be presented. Inter-rater concordance in thematic coding will be computed. A word cloud will be created with the most used nouns from the eligible posts. Verbatim quotes will be shown to illustrate themes depicted in the sample. The quantitative and qualitative analyses will be conducted separately for the posts related to kratom use during pregnancy and breastfeeding. DISCUSSION/SIGNIFICANCE: These findings could assist clinicians in identifying questions that obstetric patients may have regarding the perinatal use of this emerging substance of concern. Further research is needed to validate these findings using other social media data, such as Twitter.

OBJECTIVES/GOALS: Fetal glucose dynamics mediate many of the adverse outcomes seen in infants of diabetic mothers (IDM). The goals of this study are to identify: (1) rates of blood glucose change in normoglycemic and hypoglycemic IDM; (2) their relation to in-utero insulin exposure; and (3) their transcriptional impacts on placental and umbilical vasculature. METHODS/STUDY POPULATION: Using a longitudinal prospective study design, placental/umbilical cord tissue and maternal hemoglobin A1c (HbA1c) are being collected from mothers diagnosed with Type 1, Type 2, or gestational diabetes mellitus. Blood glucose levels are also collected from their infants at birth, and every 3-4 hours for up to 9 hours to determine the rate of change. Linear regression modeling will be used to determine associations between placental and umbilical endothelial RNA expression, umbilical cord insulin levels, and maternal HbA1c within each diabetic sub-type. Gene expression from endothelial specimens will be compared between diabetic sub-types and between normoglycemic and hypoglycemic infants via paired t-tests using Benjamini-Hochberg procedure for false discovery rate correction. RESULTS/ANTICIPATED RESULTS: We hypothesize the following; (1) glucose levels will have a steeper rate of change in hypoglycemic infants; (2) maternal HbA1c and in-utero insulin levels will correlate with the level of transcriptional change identified in placental and umbilical endothelial samples; (3) a negative association will exist between cord insulin levels and the rate of change in infant glucose levels; and (4) a positive association will exist between cord insulin level and transcriptional change on the placental and umbilical endothelium. DISCUSSION/SIGNIFICANCE: Identifying gene expression changes in diabetic placental/umbilical endothelium, and the role of insulin/glucose in these changes, is key to managing diabetic vasculopathy and its adverse outcomes. Understanding infant insulin response may also guide management of hypoglycemia and decrease the risk for neonatal intensive care unit admission.

OBJECTIVES/SPECIFIC AIMS: 1. To evaluate the effects of masculinizing chest reconstruction on the quality of life of female-to-male transgender individuals. 2. To develop and validate a quality of life survey for female-to-male transgender individuals undergoing masculinizing chest reconstruction gender-affirming surgery. METHODS/STUDY POPULATION:. We developed and refined the GENDER-Q using focus groups and exploratory interviews with FTM individuals that were recorded, transcribed, and coded. All consenting FTM patients undergoing chest reconstruction at UCSF Parnassus Medical center between 2017-2019 who meet World Professional Association for Transgender Health (WPATH) criteria for gender dysphoria will be enrolled in the study. The GENDER-Q will be co-administered with the WHO Quality of Life-BREF survey pre-operatively, 6 weeks post-operatively, and one year post-operatively through REDCAP, an online survey database. RESULTS/ANTICIPATED RESULTS: Approximately 70 patients will be enrolled in the study. Preliminary results detect statistically significant mean quality of life post-operative improvements in all three sections (Physical Health, Gender Presentation, Psychological Health) of the GENDER-Q at 6-week follow up that is maintained at one-year (p< 0.005). Statistically significant improvements were similarly achieved and maintained with the WHO QOL-BREF survey (p< 0.05). The mean reported improvement from baseline to follow-up appears to be more pronounced in the GENDER-Q survey compared to WHO QOL-BREF. The median time to complete the pre and post-operative surveys was 10 minutes. Calculation of Cronbach’s (0.67-0.81) and the Pearson Correlation Coefficient for each section revealed excellent internal validity. DISCUSSION/SIGNIFICANCE OF IMPACT: There are few studies assessing quality of life outcomes in transgender patients undergoing gender-affirming surgeries. A standardized and validated assessment tool will provide the means for which data can be pooled in large multi-center studies. Providing further evidence to support the positive health outcomes of gender-affirming surgeries can lead to broader access and reduce healthcare disparities among transgender populations.

ABSTRACT IMPACT: Limited research has been conducted on the survival of men with castration-resistance prostate cancer (CRPC) with a pre-existing history of cardiovascular disease, receiving oral androgen signaling inhibitors. This study highlights all-cause and prostate cancer-specific mortality for elderly patients with CRPC with pre-existing history of cardiovascular disease. OBJECTIVES/GOALS: Inadequate knowledge is known about the survival of men with castration-resistance prostate cancer (CRPC) with pre-existing history of cardiovascular disease (CVD), receiving oral androgen signaling inhibitors (OASI). We compared all-cause and prostate cancer-specific mortality for elderly patients with CRPC with pre-existing history of CVD. METHODS/STUDY POPULATION: An active comparator, new user design, was used to identify 2,608 men older than age 65 years with CRPC using the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database from 2011 to 2015. Patients were grouped into two analytical cohorts by CVD history. Within each analytical cohort patients were divided into two arms based on their new-user status (OASI vs. chemotherapy). All demographics and clinical characteristics were adjusted by inverse probability treatment weights (IPTWs). Unadjusted and IPTW-adjusted time-dependent Cox models, and Fine and Gray’s models were conducted to evaluate associations between OASI and all-cause and prostate cancer-specific mortality. RESULTS/ANTICIPATED RESULTS: Nearly 64.5% of patients had pre-existing CVD. We observed a lower all-cause mortality in the pre-existing CVD cohort compared to the no pre-existing CVD cohort (IPTW-adjusted hazard ratio [AHR], 0.59; 95% Confidence Interval [CI], 0.54 to 0.64; IPTW-AHR, 0.68; 95% CI, 0.59 to 0.78, respectively). Similarly, the prostate cancer specific-mortality was showed to be lower in the pre-existing CVD cohort compared to the no pre-existing CVD cohort when comparing OASI versus chemotherapy by the IPTW-adjusted time-dependent Fine and Gray’s models (IPTW-AHR, 0.60; 95% CI, 0.55 to 0.66; IPTW-AHR, 0.68; 95% CI, 0.59 to 0.80, respectively). DISCUSSION/SIGNIFICANCE OF FINDINGS: OASI showed a significant protective effect against all-cause and prostate cancer-specific mortality compared with chemotherapy; however, were less protective among patients without pre-existing CVD. Further studies are needed to investigate OASI in patients with and without pre-existing CVD.

OBJECTIVES/GOALS: Agitation has high prevalence in the emergency department (ED), but limited evidence exists regarding clinical decisions to use sedatives and physical restraint. We examined clinical factors and agitation attributes impacting thresholds for sedative and restraint use in the emergency setting. METHODS/STUDY POPULATION: We conducted a prospective cohort study of adult patients (318 yo) with acute or escalating agitation during their ED visit at an urban tertiary care referral center. Consecutive patients requiring security presence or scoring >1 on an agitation scale were enrolled during randomized 8-h blocks. We recorded patient characteristics, staff/team factors, and environmental/systems data as well as scores on 3 validated agitation scales: Agitated Behavior Scale, Overt Aggression Scale, and Severity Scale. We performed descriptive analyses, bivariable analyses, and logistic regression modeling of factors with relation to sedative/restraint use. We observed 95 agitation events on unique patients over 2 months. RESULTS/ANTICIPATED RESULTS: Median age was 42, and 62.1% were male. Most frequent chief complaints were alcohol/drug use (37.9%) and psychiatric (23.2%). Majority of events (73.7%) were associated with sedative/restraint use. Factors related to treatment course or staff interactions were the primary reasons for agitation in 56.8% of events. A logistic regression model found no association between demographics and odds of sedative/restraint use. Overt Aggression Scale scores were associated with significantly higher odds of sedative use (AOR 1.62 [1.13–2.32]), while Severity Scale scores had significantly higher odds of restraint use (AOR 1.39 [1.12–1.73]) but significantly lower odds of sedative use (AOR 0.79 [0.64–0.98]). DISCUSSION/SIGNIFICANCE OF IMPACT: External factors may be important targets for behavioral techniques in ED agitation management. Further study of the Severity Scale may allow for earlier detection of agitation and identify causal links between agitation severity and use of sedatives and restraints.

Objectives/Goals: To probe microbe and bacterial extracellular vesicle (bEV) mobility through biological barriers, we use novel multiple-particle tracking technology. The goal is to evaluate changes caused by extracellular vesicles relevant to placental function and neonatal development. Methods/Study Population: We conducted multiple particle tracking to assess whole bacterial and bEV mobility in cervicovaginal mucus. To accomplish this, cervicovaginal mucus was self-collected from 10 women. Mucus samples were characterized via wet mount, Nugent score, and pH measurements. In parallel, we cultured commercially available vaginal bacteria strains in anaerobic conditions. We isolated bEVs via ultracentrifugation, and subsequently characterized them via nanoparticle tracking analysis to measure size, ζ-potential, and concentration. We investigated reproductive tract tissues response to bEVs. We dosed vaginal, endometrial, myometrial, and placental cells lines with bEVs over a 24 h period and determined uptake, viability, and cytokine production. One-way analysis of variance was used for statistical analysis. Results/Anticipated Results: Based on our previous work, size and ζ-potential greatly affect particle mobility in mucus. G. vaginalis and M. mulieris were smaller than L. crispatus and L. iners. G. vaginalis had a more net-neutral ζ-potential compared to other bEVs. During multiple-particle tracking analysis, whole bacteria were unable to diffuse through vaginal mucus, while bEVs showed increased mobility. Through fluorescence levels, we determined M. mulieris bEVs reach >90% uptake at 24 h. Uptake was verified via microscopy. Across all strains, bEVs were not detrimental to placental viability. When investigating cytokine production in placental cells, an increase in IL-6 was seen after treatment with L. iners bEVs, while TNFα was increased after treatment with G. vaginalis bEVs. Discussion/Significance of Impact: Vaginal microbiome dysbiosis increases adverse obstetric indications. We demonstrate that bacteria are unable to ascend to reproductive tissues. We propose that bEVs travel through vaginal mucus, facilitating microbe–host communication. This impacts obstetric disease pathology and is relevant for diagnostic criteria during pregnancy.

OBJECTIVES/GOALS: Combined androgen deprivation therapy (ADT) and radiation therapy (RT) is the standard of care treatment for non-metastatic prostate cancer (NMPC). Despite the efficacy, treatment-related symptoms including fatigue greatly reduce the quality of life of cancer patients. The goal of the study is to examine the influence of combined ADT/RT on fatigue and understand its underlying mechanisms. METHODS/STUDY POPULATION: Sixty-four participants with NMPC were enrolled. Fatigue was assessed using the Functional Assessment of Cancer Therapy–Fatigue. Mitochondrial function parameters were measured as oxygen consumption from peripheral blood mononuclear cells (PBMCs) extracted from participants’ whole blood. An ADT/RT-induced fatigue mouse model was developed with fatigue measured as a reduction in voluntary wheel-running activity (VWRA) in 54 mice. Mitochondrial function was assessed in the ADT/RT mouse brains using Western blot analysis of Glucose transporter 4 (GLUT4) and transcription factor A, mitochondrial (TFAM). RESULTS/ANTICIPATED RESULTS: Fatigue in the ADT group was exacerbated during RT compared to the non-ADT group. This effect was specific to fatigue, as depressive symptoms were unaffected. PBMCs of fatigued subjects exhibited decreased ATP coupling efficiency compared to non-fatigued subjects, indicative of mitochondrial dysfunction. The ADT/RT mice demonstrated a synergistic effect of ADT and RT in decreasing VWRA. Brain tissues of ADT/RT mice exhibited decreased levels of GLUT4 and TFAM suggesting that impaired neuronal metabolic homeostasis may contribute to fatigue pathogenesis. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings suggest that fatigue induced by ADT/RT may be attributable to mitochondrial dysfunction both peripherally and in the central nervous system (CNS). The synergistic effect of ADT/RT is behaviorally reproducible in a mouse model, and its mechanism may be related to bioenergetics in the CNS.