OBJECTIVES/GOALS: We aimed to find the histology-specific markers that were predictive of post-operative long-term chordoma recurrence (≤1 year) using trained multiple tree-based machine learning (ML) algorithms. METHODS/STUDY POPULATION: We reviewed the records of patients who had treatment for clival and spinal chordomas between January 2017 and June 2021 across the Mayo Clinic enterprise (Minnesota, Florida, and Arizona). Patients were excluded if they had no histopathology or recurrence as an outcome. Demographics, type of treatment, clinical and radiological follow-up duration, histopathology, and other relevant clinical factors were abstracted from each patient record. Decision tree and random forest classifiers were trained and tested to predict the long-term recurrence based on unseen data using an 80/20 split. The performance of the model was optimized using tenfold cross-validation. RESULTS/ANTICIPATED RESULTS: One hundred fifty-one patients were identified: 58 chordomas from the clivus, 48 chordomas of the mobile spine, and 45 sacrococcygeal. Subtotal Resection followed by radiation therapy, was the most common treatment modality, followed by Gross Total resection, then radiation therapy. The multivariate analysis defines the molecular predictors of recurrence following resection. S100 and pan-cytokeratin is more likely to increase the risk of post-operative recurrence (OR= 3.67; CI= [1.09,12.42], p=0.03). In the decision tree analysis, a clinical follow-up > 1897 days was found in 37 % of encounters and a 90% chance of being classified for recurrence (Accuracy= 77%). Factors predicting long-term recurrence are the patient’s age, type of surgical treatment, location of the tumor, S100, pan-cytokeratin, and EMA. DISCUSSION/SIGNIFICANCE: Our molecular and clinicopathological variables combined with tree-based ML tools successfully demonstrate a high capacity to individually identify the patient’s recurrence pattern with an accuracy of 77%. S100, pan-cytokeratin, and EMA were the histologic drivers of recurrence.

OBJECTIVES/GOALS: 1. Describe the importance of community engaged research. 2. Explore the impact of participating in Community Engagement Studios from the perspective of the community expert. 3. Explain how Community Engagement Studios can positively impact clinical and translational sciences. METHODS/STUDY POPULATION: Stakeholders who participate in CE Studios are defined as community experts (CEs); care-givers, patients, researchers, etc. They are not classified as research participants, but rather consultants, who are designated with the task of providing input on the specified research study. CEs were recruited in a variety of formats such as clinics, social media, and other community-based settings. To be eligible for the study, CEs had to participate in at least one CE Studio prior to data collection. Data was collected from an online survey from April to September of 2019. One hundred and fifteen CEs participated in the study. We conducted thematic analysis on a total of 233 participant responses to three open-ended questions assessing attitudes towards research, desire to engage in research, and understanding of research. RESULTS/ANTICIPATED RESULTS: Ten major themes emerged from the data: involvement, togetherness, trust, value, confidence, community engagement, community connectedness, encouraging others to participate, increase knowledge and awareness, and respect. A major overarching theme of inclusion was presented in the data. Findings from this study indicate that CE Studios provide a space for CEs to gain a better understanding of the multi-faceted research process, offer insight into ways to include historically excluded populations, and increase trust, confidence and respect for researchers and the research process. Moreover, community experts felt connected to their community by participating in CE Studios and expressed interest in encouraging others to participate. DISCUSSION/SIGNIFICANCE: CEs are a crucial component of the research process, and their experiences should be utilized and reflected upon as we work to reduce health disparities. CE Studios serve to build trust, identify strategic areas of improvement in research, and provide a foundation to introduce community members to the multiple ways to engage in research.

ABSTRACT IMPACT: Pharmacological activation of KATP channels may provide analgesia and attenuate opioid tolerance and withdrawal OBJECTIVES/GOALS: Our long term goal is to develop therapeutics for the treatment of the overuse of opioids. The objective of this application is to test novel KATP channel-targeting prodrugs in rodent models of neuropathic and inflammatory pain in addition to opioid tolerance after chronic morphine administration. METHODS/STUDY POPULATION: In one study, two different measures for chronic pain were implemented in mice. Male and female mice (n=10) were subjected to spinal nerve ligation (SNL) or intraplantar injection of Complete Freund’s Adjuvant (CFA) to induce neuropathic and inflammatory pain, respectively. Administration of KATP channel prodrugs (60ug, it) attenuated mechanical hypersensitivity after SNL or CFA compared to vehicle (saline). In a separate study, changes in mechanical hypersensitivity were tested while mice undergo chronic morphine treatment (15mg/kg, 2x, 5 days) with administration of the prodrugs. Tolerance was measured as the loss of antinociception, and withdrawal is measured ˜24 hours after the final morphine injection. RESULTS/ANTICIPATED RESULTS: Intrathecal administration of either KATP channel prodrugs significantly attenuated mechanical hypersensitivity after SNL and significantly attenuated mechanical hypersensitivity after CFA in mice. We predict that intrathecal administration of these prodrugs will also attenuate morphine tolerance and withdrawal in mice. This hypothesis is based off our previous data indicating non-water soluble KATP channel agonists produce analgesia and attenuate morphine tolerance in mice. DISCUSSION/SIGNIFICANCE OF FINDINGS: Pharmaceutical strategies to utilize KATP channels for therapeutics have been hindered due to the low solubility and low ability to cross the neurovascular unit. Newly developed, water-soluble KATP channel openers could be useful pharmaceutical strategy to reduce chronic pain, opioid tolerance, and withdrawal in human populations.

OBJECTIVES/SPECIFIC AIMS: The Clinical and Translational Science Award (CTSA) program is a national consortium of 50+ academic medical research centers charged with accelerating the translation of clinical research. In 2017, the NIH National Center for Advancing Translational Sciences anticipates total CTSA program funding of over $500M. The consortium’s hub-and-spoke structure makes it a natural dissemination network, and the newest funding announcement makes dissemination of innovation across the consortium an explicit goal, but characteristics of CTSA hubs as adopters and transmitters of innovation are unknown. METHODS/STUDY POPULATION: A content analysis was conducted using data from CTSA hub Web sites (n=64) and a structured coding taxonomy based on 6 constructs drawn from literature about diffusion of innovation in service organizations (Greenhalgh et al., 2004): dissemination priority, institutional complexity, communication infrastructure, support for dissemination/implementation functions, cross-institutional collaboration/networking, and leadership composition. RESULTS/ANTICIPATED RESULTS: In total, 52% of hubs will renew under the new PAR in the next few years, providing an incentive to demonstrate dissemination capacity (although hubs will likely lag in operationalizing these activities until they are funded). A third of hubs (34%) represent more than one academic/research institution, and almost 80% of hubs have more than one clinical affiliate. To accommodate these different levels of institutional complexity, broad diffusion will require multi-modal, locally adapted dissemination efforts. Only 25% of hubs have capacity to undertake additional dissemination activities, and only 27% provide formal D&I support, suggesting that additional capacity/support will be needed to operationalize the CTSA dissemination mission. In total, 30% of hubs participate in cross-institutional collaboration/networking, so many may not have existing norms/tools supporting inter-institutional collaboration, but 77% include leadership from outside the School of Medicine, facilitating effective intrainstitutional dissemination. DISCUSSION/SIGNIFICANCE OF IMPACT: Understanding more about CTSA hubs as both adopters and transmitters of innovation can facilitate strategic use of these sites as a built-in dissemination network to amplify the reach and impact of clinical innovation and improve population health. Based on this initial analysis, the CTSA network does not appear to be fully primed for broad, rapid dissemination of innovation across its sites. In-depth interviews are being conducted to investigate CTSA hubs’ perceptions of their dissemination capacity and roles as adopters and transmitters of innovation.

OBJECTIVES/SPECIFIC AIMS: The objectives of this study are to determine whether high-frequency ipsi-lesion or low-frequency contra-lesion ECS improves forelimb function following experimental stroke in aged animals with focal and large strokes. We also want to investigate whether ECS-induced improvements in motor function are related to an enhancement of neural structural plasticity (dendrites and synapses) and changes in growth promoting (BDNF) and growth inhibiting (NOGO-A) expression in the infarcted motor cortex in young and aged animals. METHODS/STUDY POPULATION: We will investigate whether excitatory ECS of the infarcted cortex or inhibition of the noninfarcted cortex combined with daily impaired-forelimb rehabilitative training (RT) results in greater motor functional recovery compared to RT alone. Immunohistochemical (IHC) analyses and unbiased stereological techniques will be performed to investigate changes in proteins associated with dendritic restructuring (MAP2), synaptic plasticity (PSD95 and synaptophysin), and alteration in the expression of BDNF and NOGO-A. RESULTS/ANTICIPATED RESULTS: We expect that inhibitory ECS of the noninfarcted motor cortex will improve behavioral outcomes in moderate to severe stroke animals compared with excitatory ECS or no stimulation (RT alone) animals. We predict that the ECS condition that improves motor performance most significantly compared with RT alone will have a corresponding greater increase in remaining ipsi-infarct motor cortical dendritic and synaptic plasticity (demonstrated by a greater density of MAP2, synaptophysin, and PSD-95 immunoreactivity), and greater expression of BDNF. It is unknown, but also expected that better behavioral recovery will coincide with a greater reduction in NOGO-A in the injured motor cortex. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies will aid in creating a model that will allow for a better understanding of the relationship between brain stimulation, severity of injury and, in future studies, aging. These studies will also help clarify previous conflicting brain stimulation results.

OBJECTIVES/SPECIFIC AIMS: The cellular mechanisms that underlie the IFNβ/α ratio that predicts response are not known. Effects of IFN on single immune cells may be masked in whole blood or mixed cell populations. By studying the effect of IFNβ/α activity ratio on individual monocytes, we can determine the functional impact of the IFN ratio and suggest the cellular mechanisms that underlie response/non-response to TNFi therapy in RA. METHODS/STUDY POPULATION: We used single cell analysis to investigate whether monocyte gene expression differs significantly between RA patients according to their pre-TNFi serum IFN-β/α ratio. Single classical (CL) and non-classical (NC) blood-derived monocytes were isolated from 15 seropositive RA subjects prior to biologic therapy. Subjects were grouped by pre-TNFi serum IFN-β/α ratio into two groups, those with a high IFN-β/α ratio (≥1.3, n = 6) and those with a low IFN-β/α ratio (<1.3, n = 9). 87 target genes were analyzed. Genes that varied significantly between the groups by categorical analyses were tested in multivariate logistic regression models. RESULTS/ANTICIPATED RESULTS: Every participant was seropositive for rheumatoid factor and antibodies to cyclic citrullinated peptide. Among the participants in the groups, there were no significant differences in age or DAS scores (P>0.05). The treatments were comparable and none were being treated with biologic therapy. There were striking differences in monocyte gene expression between patients with pre-treatment blood IFNβ/α activity <1.3 and ≥1.3. Expression of (1) key type I IFN pathway genes (JAK1, STAT2, IFIT2, IFIH1, PRDM1); (2) IL12; (3) CD36; and (4) CTLA4 were the strongest differentiators between groups (p<0.0001 for each, corrected for multiple comparisons). DISCUSSION/SIGNIFICANCE OF IMPACT: In this study we were able to measure gene expression in single monocytes from seropositive RA patients prior to biologic treatment. Within-cell co-expression patterns demonstrate biological differences in monocytes of RA patients with an IFNβ/α ≥1.3, the ratio of type I IFNs which predicts non-response to TNFi. The data suggest that there may be differential IFN production and pathway activation in patients who do not respond to TNFi. The increased expression of CD36 in monocytes from RA patients with high IFN β/α activity may be a reflection of increased “foam cells” in the inflamed tissue of patients who do not respond to TNFi. Enrichment of CTLA4 in those with high serum IFNβ/α suggests that CTLA4-Ig may be less likely to be an effective alternative for someone who is not likely to respond to TNFi. Current work includes determining whether the peripheral blood findings reflect altered cellular composition, type I IFN production and signaling in the synovium. Significance: This work will help to develop a more individualized approach to therapy in RA and determine an immunological basis of response/non-response to TNFi.

ABSTRACT IMPACT: We aim to understand how LTCCs impact cerebellar function. OBJECTIVES/GOALS: L-type calcium channels (LTCCs) are important in activity-dependent neurite outgrowth, which comprises neurite initiation and elongation. We used cerebellar granule neurons (CGNs) to differentiate between LTCC effects on neurite initiation vs elongation. We also tested cerebellar function in mice lacking specific LTCCs with behavioral assays. METHODS/STUDY POPULATION: CGNs were cultured from 129SvEv mouse pups at P4-P6. Potassum chloride (50mM) was used to stimulate neuronal cultures for 24 hours. Isradipine (20nM) was added to culture medium to inhibit all LTCCs for 1 hour. For Cav1.2 deletion, we crossed Cav1.2 conditional knockout mice (Cav1.2-cKO) to Syn-Cre mice (for deletion in most neurons) or Atoh1-Cre mice (for deletion in CGNs). The Cav1.2-cKO line was maintained on a 129SvEv background. For constitutive Cav1.3 deletion, mice were maintained on a C57BL/6NTac. Behavioral tasks included open field, rotarod, and Erasmus Ladder. Data were analyzed with sexes combined and separated to assess for sex as a biological variable. Studies were analyzed by one-way ANOVA, two-way ANOVA, or generalized linear mixed model, where appropriate. RESULTS/ANTICIPATED RESULTS: CGNs exhibited an increase in neurite initiation but not elongation when stimulated with potassium chloride, consistent with previous reports of activity-dependent neurite outgrowth in this cell type. LTCC inhibition with isradipine blunted KCl-induced neurite initiation. We observed no change in the length of either primary or secondary neurites with isradipine treatment with or without KCl stimulation. In our behavioral experiments, we observed no deficits in open field, rotarod, or Erasmus Ladder when Cav1.2 was deleted in most neurons (driven by Syn-Cre expression) or in cerebellar granule neurons (driven by Atoh1-Cre expression). In contrast, loss of Cav1.3 was associated with impaired motor learning in the rotarod task without evidence of ataxia on Erasmus Ladder. DISCUSSION/SIGNIFICANCE OF FINDINGS: We show a specific role for LTCCs in activity-dependent CGN neurite initiation. While loss of Cav1.2 does not affect motor learning, loss of Cav1.3 does impair motor learning. Our results help expand our understanding of LTCC function in cerebellar neurodevelopment and function.

OBJECTIVES/GOALS: Fatigue is a distressing side effect of cancer and its treatment. It is a subjective symptom that can include mental, physical, emotional, and motivational components. We sought to determine whether preventing inflammation affects fatigue-like behavior in a mouse model of radiation therapy. METHODS/STUDY POPULATION: C57BL/6 mice received three consecutive 8 gray doses of daily peripheral irradiation. We used voluntary wheel running activity to measure fatigue-like behavior before and after this period. Minocycline, an antibiotic with anti-inflammatory effects, was administered beginning a week before irradiation and continued throughout the experiment. We also tested mice lacking the toll-like receptor adaptor protein, MyD88. Cognitive abilities were tested using spontaneous alternation in a Y-maze. RESULTS/ANTICIPATED RESULTS: We found that minocycline reduces fatigue-like behavior exhibited after irradiation, but had no effect on pre-irradiation activity levels. Similarly, fatigue-like behavior after radiation was partially reversed by genetic loss of MyD88. Y-maze spontaneous alternation performance remained similar in all groups. DISCUSSION/SIGNIFICANCE OF IMPACT: Both pharmacological and genetic anti-inflammatory manipulations increased voluntary activity levels after irradiation. Our results suggest that inflammation is an important factor in the development of fatigue-like behavior. Modulators of inflammatory processes hold potential for alleviating fatigue.

OBJECTIVES/GOALS: Frequent utilizers of emergency departments (ED) make up a substantial share of overall ED use. Within pediatric emergency departments (PED), infants represent an age group that make up a disproportionate share of PED visits. The objective was to compare patterns of PED use for children less than 1 year of age by visit frequency and resource utilization. METHODS/STUDY POPULATION: Retrospective cohort study of infants less than 1 year presenting to 5 EDs in one health system over a 5-year period, with a 365-day follow-up after each index visit. Patient characteristics (age, sex, race/ethnicity, presence of chronic condition) and visit characteristics (arrival day/time, acuity level, disposition, testing (labs and radiographs, medications) were assessed. The relationship between patient and visit characteristics with utilization and repeat visits was assessed using multivariable regression. RESULTS/ANTICIPATED RESULTS: A total of 20,620 patients with 33,127 ED visits during study timeframe. Thirty three percent (n=6842) had more than one visit in a year; 3964 (19.2%) had two visits, 1542 (7.5%) had three visits, and 1336 (6.5%) had 4 or more visits. Across all visits, over half (52%) were low acuity. The most common diagnoses were respiratory diseases (27%), systemic states (including fever, viral illness, 23%), and gastrointestinal diseases (15%). These diagnoses remained the most common for those with 1, 2, 3, and ≥ 4 ED visits during follow up. As ED visit frequency increased, there was an increase in percentage of children who were older, non-Hispanic Black, and triaged as low acuity. Infants with ≥ 4 ED visits were more likely to be without a chronic condition, have no medications or testing ordered, and be discharged. DISCUSSION/SIGNIFICANCE: There was high ED utilization for those without chronic conditions who were least likely to need medications, testing, and hospital admission. With increasing attention paid to high-utilization in healthcare, it is important to assess why infants use the ED at high rates and develop systems to improve high value care while decreasing resource burden.

As the pace of biomedical innovation rapidly evolves, there is a need to train researchers to understand regulatory science challenges associated with clinical translation. We describe a pilot course aimed at addressing this need delivered jointly through the Mayo Clinic Center for Clinical and Translational Science and the Yale-Mayo Center for Excellence in Regulatory Science and Innovation. Course design was informed by the Association for Clinical and Translational Science’s Regulatory Science Working Group’s competencies. The course used didactic, case-, and problem-based learning sessions to expose students to regulatory science concepts. Course evaluation focused on student satisfaction and learning. A total of 25 students enrolled in the first two course deliveries. Students represented several disciplines and career stages, from predoctoral to faculty. Students reported learning “an incredible amount” (7/19, 36.8%) or “a lot” (9/19, 47.4%); this was reflected in individual coursework and their course evaluations. Qualitative feedback indicated that assignments that challenged them to apply the content to their own research were appreciated. The heterogeneity of students enrolled, coupled with assessments and course evaluations, supports the statement that there is a growing need and desire for regulatory science-focused curricula. Future research will determine the long-term impact.

Developing the translational research workforce is a goal established by the National Center for Advancing Translational Science for its network of Clinical and Translational Science Award Program hubs. We surveyed faculty and research staff at our institution about their needs and preferences, utilization of existing trainings, and barriers and facilitators to research training. A total of 545 (21.9%) faculty and staff responded to the survey and rated grant development, research project development, and professional development among their top areas for further training. Faculty prioritized statistical methods and dissemination and implementation, while staff prioritized research compliance and research administration. Faculty (73.9%; n = 119) and staff (87.3%; n = 165) reported that additional training would give them more confidence in completing their job responsibilities. Time and lack of awareness were the most common barriers to training. Our results indicate the value of training across a range of topics with unique needs for faculty and staff. This pre-COVID survey identified time, awareness, and access to training opportunities as key barriers for faculty and staff. The shift to remote work spurred by the pandemic has further heightened the need for effective and readily accessible online trainings to enable continuous development of the clinical and translational research workforce.

There is a growing trend for studies run by academic and nonprofit organizations to have regulatory submission requirements. As a result, there is greater reliance on REDCap, an electronic data capture (EDC) widely used by researchers in these organizations. This paper discusses the development and implementation of the Rapid Validation Process (RVP) developed by the REDCap Consortium, aimed at enhancing regulatory compliance and operational efficiency in response to the dynamic demands of modern clinical research. The RVP introduces a structured validation approach that categorizes REDCap functionalities, develops targeted validation tests, and applies structured and standardized testing syntax. This approach ensures that REDCap can meet regulatory standards while maintaining flexibility to adapt to new challenges. Results from the application of the RVP on recent successive REDCap software version releases illustrate significant improvements in testing efficiency and process optimization, demonstrating the project’s success in setting new benchmarks for EDC system validation. The project’s community-driven responsibility model fosters collaboration and knowledge sharing and enhances the overall resilience and adaptability of REDCap. As REDCap continues to evolve based on feedback from clinical trialists, the RVP ensures that REDCap remains a reliable and compliant tool, ready to meet regulatory and future operational challenges.

OBJECTIVES/GOALS: Decoding the origins of cell-free DNA (cfDNA) released from dying cells in a liquid biopsy sample (e.g. blood) can provide insight into the dynamic, organism-wide changes reflective of health and disease state. Making cfDNA an ideal target for genomic monitoring of disease-related changes. METHODS/STUDY POPULATION: Methylome-wide sequencing (WGBS) data present unique statistical challenges. To this end, we developed a novel statistical method using an Expectation-Maximization algorithm to decode the cellular origins of cfDNA fragments in liquid biopsies. Our flexible, probabilistic method leverages the co-regulation of neighboring CpG sites on the individual sequencing read to facilitate tissue of origin analysis, as opposed to prior methods that focus on the methylation rate of a single CpG site. We assess the performance of our model in various simulated settings and apply our model to an important clinical example in which we are able to detect early off-target tissue damage from radiation therapy via minimally invasive blood draws. RESULTS/ANTICIPATED RESULTS: We found our model more effective at capturing the range of biologically plausible methylation patterns on cfDNA read fragments compared to prior models that use single CpG sites. We also show our model is robust to high levels of noise inherent with WGBS data. We demonstrate the accuracy of cell-type proportion estimation on in-silico mixed cfDNA samples from real WGBS data. Finally, we use our model in a clinical application. We detect significant (p < 0.05) increases in cellular contributions from lung and cardiac tissue in breast cancer patients (n=15) undergoing radiation therapy compared to baseline. We also detect novel signals of radiation induced toxicity to the liver in right-sided breast cancer patients (n=8) receiving radiation treatment compared to matched left-sided breast cancer patients (n=7). DISCUSSION/SIGNIFICANCE: Here we address an unmet need in developing novel statistical methodologies that can handle the unique complexities of methylated cfDNA obtained from liquid biopsy samples. We also demonstrate the far-ranging clinical utility of serial liquid biopsy sampling to complement and advance standards of clinical care in oncology and other pathologies.

OBJECTIVES/SPECIFIC AIMS: This project aims to determine the key molecular pathways that link increased myocardial wall stress to cardiomyocyte hypertrophy and subsequent heart failure. We will use a cardiac microtissue (CMT) model with dynamically tunable cantilever stiffness to examine changes in CMT hypertrophy and electro-mechanical properties in response to increased afterload (cantilever stiffness). Subsequently, we will determine if inhibition of pro-hypertrophic or anti-hypertrophic pathways alter the hypertrophic response to increased afterload. Primary outcomes for this study are static/dynamic force, minimum electric field strength (VT), maximum capture rate (MCR), average cell area, and tissue cross-sectional thickness, and secondary outcomes are degree of myoblast activation and apoptosis. METHODS/STUDY POPULATION: CMT platforms will be fabricated using iron-doped polydimethylsiloxane (PDMS) to create magnetically tunable cantilevers. Cantilever stiffness will be increased with the application of an external magnetic field. Cantilever stiffness will be measured using a capacitance probe, where the force required to deflect both the cantilever and calibration probe is in accordance with Hooke’s Law. Human induced pluripotent stem cell cardiomyocyte (hiPSC-CMs) will be cultured and matured as 3D CMTs. In-vitro static/dynamic force generation will also be calculated by measuring the deflection of the cantilevers and applying Hooke’s law. CMTs will be paced using carbon electrodes to obtain VT and MCR. Structural data will be obtained using immunostaining and confocal microscopy. Finally, we will use pharmacologic inhibitors to inhibit molecular pathways that we identified in prior genetic screens such as ABCC8 (anti-hypertrophic mediator) and C1QTNF9 (pro-hypertrophic mediator). We will examine each of these pathways in low- and high-stiffness conditions. RESULTS/ANTICIPATED RESULTS: We believe increased afterload will cause significant hypertrophy, measured by increases in CMT cross-sectional thickness, cardiac myocyte area, myofibroblast activation, and myocyte apoptosis. In addition, we expect to see increases in static/dynamic force, increased voltage threshold, and decreased maximum capture rate. Preliminary results show a 64.3% increase in force generation when stiffness is increased by approximately 30%, and a 44.4% decrease in force generation when stiffness is decreased by approximately 30%. Finally, we expect that inhibiting a pro- or anti-hypertrophic molecular pathway will weaken or strengthen the hypertrophic response to increased afterload, respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: To our knowledge, our lab is the first to create a dynamically tunable afterload system in the cantilever CMT model. This advance provides us with a robust platform to determine the molecular pathways that cause increased myocardial wall stress to result in cardiomyocyte hypertrophy and heart failure, which remain a critical knowledge gap in our understanding of cardiovascular disease. With more precise understanding of these pathways, we will equip ourselves with the knowledge to develop novel therapeutic agents to prevent the development or progression of heart failure.

OBJECTIVES/GOALS: Fertility preservation (FP) allows transgender and gender diverse (TGD) patients undergoing gender-affirming therapy to pursue genetic parenthood. Barriers to care exist leading to its underutilization; while these barriers to care have been investigated, the true utilization rates of FP services by TGD patients are unknown. METHODS/STUDY POPULATION: This study includes a retrospective chart review of adult and pediatric patients from Mayo Clinics Transgender and Intersex Specialty Care Clinic who have provided research authorization. We will assess if FP was discussed and review if the patients proceeded to make appointments with either the Department of Reproductive Endocrinology & Infertility or the Department of Urology. FP terms include semen cryopreservation, sperm extraction, sperm aspiration, testicular tissue cryopreservation, oocyte cryopreservation, embryo cryopreservation, and ovarian tissue cryopreservation. Patient demographic data will be collected to determine associations between utilization rate and age, sex recorded at birth, gender identity, race, ethnicity, and variables related to socioeconomic determinants of health. RESULTS/ANTICIPATED RESULTS: We hypothesize that the rate of FP utilization will be higher for patients who begin to affirm their gender at the age of 25 or older, and for those seeking feminization treatment in comparison to pre-pubescent adolescents, young adults, and transgender men. Of FP options, semen cryopreservation will be most utilized, followed by oocyte cryopreservation; both will be more utilized than embryo cryopreservation, sperm extraction, sperm aspiration, testicular tissue cryopreservation, and ovarian tissue cryopreservation. Employment, and insurance status and coverage, will play a role in FP service utilization after initial consult. This study was approved by Mayo Clinics Institutional Review Board. Data on N=611 patients is expected to be abstracted and analyzed prior to Translation 2022. DISCUSSION/SIGNIFICANCE: This study will be one of the first to examine the rate of FP utilization by TGD patients with respect to the age and timing of gender-affirming therapy initiation. Understanding the rate of utilization of FP services will allow for the creation of age-appropriate education materials for TGD patients pursuing gender affirmation and FP at Mayo Clinic.