Significant improvements have been achieved to enhance the patient-centricity of clinical research, including the development and utilization of novel clinical trial endpoints. These include endpoints that harness outcomes that are important to patients and reflect the patients’ lived experiences. This may take the form of utilizing variables such as days alive and out of hospital (DAOH) and quality-of-life adjusted outcomes. The use of composite outcomes can be used to enrich patient-centricity by weighting or ranking events. These approaches have several nuances that should be considered including selecting appropriate events, defining outcomes, how to elicit or construct weights, and whose opinions to consider. After weights have been determined, a variety of approaches exist to combine weights with outcomes and make comparisons between groups. The approaches, including the win ratio, weighted win ratio, desirability of outcome ranking (DOOR), multicriteria decision analysis (MCDA), and variations of time-to-first composite event analyses, have unique advantages and challenges depending on the clinical scenario. While improving patient-centric outcomes is of high importance to multiple stakeholders, more comparative work is needed to characterize the implications of alternative approaches.

Objectives/Goals: Large-scale tumor sequencing efforts have led to annotations of novel cancer hotspot mutations that may underlie driver or cooperative function. We have sought to define the molecular consequences of such hotspots associated with pediatric DICER1 syndrome cancers, with the ultimate goal of revealing novel targets that may inform new standards of care. Methods/Study Population: We have performed genomic analysis to identify tumor types (in TCGA and MSK-IMPACT patient data) for which mutations in the Dicer1 gene (encoding Dicer protein) emerge as the dominant signature of driver function. As Dicer is a critical RNA processing factor responsible for the generation of microRNAs, which are posttranscriptional gene regulatory molecules, we have modeled these mutations in human embryonic stem cells in order to study the direct effects on miRNAs and their target genes in an isogenic background. In addition to providing the required setting for unambiguous attribution of function to specific mutations, clonal human ES cells offer an opportunity for modeling of both developmental and cancer requirements associated with altered Dicer function. Results/Anticipated Results: Through generation of genomics and functional datasets from matched genotypes in Dicer mutated human ES cells, we have identified specific alterations in miRNAs and their effects on target genes. Unexpectedly, we found direct evidence for both loss of function and gain of function attributable to Dicer mutations. In addition, through integrated analysis of genomic data from tumor sequencing datasets and our human ES cell models, we have identified potential miRNA and target gene alterations that underlie tumorigenic potential, nominating gene candidates for targeted therapy in DICER1 syndrome. Direct mouse modeling of such candidate gene targets has revealed evidence for driver function of identified miRNA and their targets. Discussion/Significance of Impact: DICER1 syndrome cancers comprise a wide variety of rare pediatric tumor types. Presently, we still lack an effective standard of care. Furthermore, the previous lack of molecular profiling precluded targeted therapy opportunities. Our precise knock-in modeling of Dicer hotspots and deep profiling of relevant tumors now provide candidate targets.

OBJECTIVES/SPECIFIC AIMS: This is a prospective, longitudinal cohort study correlating postoperative opioid use, prescription availability at discharge and patient use at home using survey methodology and longitudinal cohort data. The primary objectives of this project are twofold. First, we will determine whether the number of opioid pills prescribed after surgery increases the risk of abuse, diversion and conversion to chronic use. Next, we will determine whether patient and parent characteristics, behavioral factors and medical comorbidities increase the risk of opioid abuse, diversion and conversion to chronic use after surgery. METHODS/STUDY POPULATION: A mixed-methods approach will be employed utilizing retrospective and prospective medical record review, survey methodology, and longitudinal cohort matching with California’s Controlled Substance Utilization Review and Evaluation System (CURES) reports of opioid usage. Surveys will be administered before and after surgery and will capture both parent and patient level factors that may influence longitudinal opioid use. Adolescents and young adults 13-20 years old discharged from Children’s Hospital Los Angeles undergoing one of seven procedures most commonly associated with an opioid prescription at time of discharge (spinal fusion, arthroscopy, bowel resection for inflammatory bowel disease, pectus excavatum repair, tonsillectomy, pilonidal excision and hip reconstruction) will be enrolled. RESULTS/ANTICIPATED RESULTS: Inpatient and outpatient opioid usage will predict incidence of diversion, abuse and conversion to chronic usage. Furthermore, socioeconomic factors and behavioral comorbidities such as anxiety and depression will be identified as predictors of diversion, abuse and conversion to chronic opioid use. DISCUSSION/SIGNIFICANCE OF IMPACT: This study will be one of the first to identify opioid prescribing variation in children who require surgery and will identify multiple provider, patient and family level factors influencing use. Furthermore, it will identify factors predictive of possible diversion, abuse and conversion to chronic use. The results of this exploratory analysis will lay the foundation for future studies utilizing physician decision support tools, aid in crafting “best practice” patterns and clinical benchmarks for opioid prescribing, and serve to inform policy makers on the most impactful ways to optimize opioid prescribing practices for adolescents who require surgery.

OBJECTIVES/SPECIFIC AIMS: Our overall objectives are to determine (i) whether natural disaster-related prenatal maternal stress (PNMS) alters infants’ effortful control (EC) at two years of age, and (ii) if the timing of exposure moderates its effects on toddlers EC. METHODS/STUDY POPULATION: We propose a longitudinal study with 50 mother-toddler dyads. Natural disaster-related PNMS would be measured at 12-15 and 18-24 months of age and will include: objective exposure and maternal distress. EC will be measured with a questionnaire and a Laboratory Temperament Assessment Battery at two years of age. RESULTS/ANTICIPATED RESULTS: We anticipate that children exposed to Hurricane-related PNMS would present low EC levels compared to those with low prenatal exposure. DISCUSSION/SIGNIFICANCE OF IMPACT: These results are expected to provide evidence for further promoting early intervention and ameliorating negative effects of PNMS on child outcomes.

OBJECTIVES/GOALS: To validate the previously developed ototoxicity prediction model for objective (i.e., audiometric-defined) hearing loss from cisplatin-based and radiation treatments in a new cohort of head and neck cancer patients treated from 2018 to 2019. METHODS/STUDY POPULATION: This study will use a cohort of 106 patients undergoing treatment for head and neck cancers at a single institution to temporally validate a model for post-treatment ototoxicity. We are interested in understanding if this model will be able to predict ototoxic risk (calibration) and if this model can differentiate high- and low-risk patients (discrimination). Observed and predicted values for audiometric hearing loss will be calculated and then compared using a calibration curve available in SAS v9.4, while the c-index (area under the receiver-operator curve) will be used to assess discrimination. The implementation of this model will be assessed in a clinical setting. RESULTS/ANTICIPATED RESULTS: The validation cohort is generally similar in age (61 years) and sex-mix (23% female) to the original cohort. However, there seems to be a different case-mix the types of treatments with more patients receiving cisplatin overall (59% vs. 43%), but fewer getting induction and high-dose cisplatin (1% vs. 13%). The original model showed good calibration and fair discrimination in the validation cohort with and area under the curve of 0.700. This concordance statistic suggests possibly-useful discrimination and the calibration curve suggests the model is well-calibrated. DISCUSSION/SIGNIFICANCE OF IMPACT: This project can improve clinical treatment paradigms, enhance patient education, and reduce healthcare costs. Our model allows oncologists to weigh the risks of hearing loss with the benefits of treatment on an individualized level before treatment, facilitating informed treatment decision-making.

OBJECTIVES/SPECIFIC AIMS: Understand the association between ADI and completion of preventative cancer screening and high risk behaviors -Identify how to operationalize ADI in clinical settings to assist care teams and improve overall care delivery. METHODS/STUDY POPULATION: Aim 1: Paneled Mayo Clinic patients living in MN, IA, or WI Aim 2: BRFSS survey respondents from MN, IA, or WI Aim. 3: Community health officials and Mayo Clinic care teams. RESULTS/ANTICIPATED RESULTS: We anticipate that areas with greater composite deprivation will have lower completion rates of cancer screening and higher risk behaviors. DISCUSSION/SIGNIFICANCE OF IMPACT: No single body of work has illustrated how ADI relates to completion of preventative cancer screening and high-risk behaviors. Due to the limited research focused on area deprivation and behavioral health, our work will identify some of the first national hot spots with high deprivation and high-risk behaviors. Additionally, this is one of the first studies describing spatial variation in health outcomes for Mayo Clinic patients. Understanding the association between ADI and patient adherence to preventative screening will allow us to support care teams in providing personalized and sustainable care for patients living in areas of high deprivation. The strength and novelty of this project is in the utility of the mixed methods design, which provides a more complete understanding of geographic disparities and a unique perspective to patient care, a perspective that is not portrayed in existing literature.

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.

OBJECTIVES/GOALS: This study’s goal is to examine the feasibility and acceptability of using VRM to impact the APP of adults in the inpatient setting. Aims include examining the: 1) feasibility of VRM for APP management; 2) acceptability of using VRM for APP management; and 3) experience of VRM for APP management. METHODS/STUDY POPULATION: To comprehensively examine participants’ experience of using VRM for APP, this study will employ a convergent mixed-methods design in which living kidney donors (N = 45) will be recruited to serially use VRM during their hospital stay. Feasibility and acceptability will be evaluated using descriptive and inferential statistics evaluating patient-reported outcome (PRO) measures taken pre-, post- and 1-hour post-VRM, PRO measures extracted from the participant’s electronic health record and data on VRM use. Semi-structured interviews will allow formulation of inferences based on participants’ experience of VRM for APP management and their insights on content, deployment, and clinical use of VRM. RESULTS/ANTICIPATED RESULTS: This in-process study expects: 1) an adequate sample of participants undergoing living kidney donor surgery who agree to enroll with retention of >90% of participants (Aim 1); 2) participants to report VRM as an acceptable and suitable treatment, feel “present” and interested in the VR environment, and feel comfortable using VRM in the hospital (Aim 2); and 3) to provide insight into participants’ experience of VRM for APP, understanding of extended VRM use for APP analgesia, examination of key variables affecting participants’ experience of VRM for APP and feedback about VRM procedures and protocol to inform future VRM use for APP management (Aim 3). DISCUSSION/SIGNIFICANCE OF IMPACT: Results of the proposed study will inform future clinical testing and deployment of VRM, guide future use of VRM as an adjunct for inpatient APP management, and provide insight into inpatients’ experience of VRM for APP analgesia.

OBJECTIVES/GOALS: Mayo Clinic (MC) launched the Rapid Activation Trial (RAT) pilot program in 2022 to expedite the activation of high priority and high impact clinical trials. The objective was to develop a process for rapid activation through robust screening, prioritization, and project management (PM) support. METHODS/STUDY POPULATION: The project team developed a robust screening and approval process for the RAT program using a combination of an objective scoring tool (based on strategic priorities) and a diverse selection committee to screen and approve eligible trials. Sponsors had to commit to RAT program timelines. Upon approval, trials were prioritized at the highest level within each business unit involved in the activation process. The number of trials approved annually were limited to 8 to manage volume and facilitate seamless prioritization with an activation timeline goal of 6 weeks. Project management support for RAT program focused on financial, regulatory, logistical, and operational elements to open trials expeditiously. RESULTS/ANTICIPATED RESULTS: In 2022, thirteen (13) applications were received and eight (8) were approved by the RAT selection committee. The approved trials activated with a median open to enrollment time of 6.4 weeks from engaging with business units. They also aligned closely with organization’s strategic priorities, including but not limited to Investigator Initiated Trials, Multi-Site protocols, IND/IDE protocols, Rare Diseases, First in Human and Commercialization potential trials. PI and study team feedback was positive. In 2023, the RAT program was renewed due to the pilot’s significant success in 2022. The goal is to open 10 trials and 5 have been activated by the end of Q3, 2023 with a median timeline of 6 weeks. DISCUSSION/SIGNIFICANCE: Rapid activation of high priority and high impact clinical trials enables an organization to strategically prioritize and open complex clinical trials. This allows the delivery of innovative, timely cures to patients in an expeditious timeline.

OBJECTIVES/GOALS: The FDA allows physicians to request clinical use of investigational drugs, biologics, and devices for patients with no satisfactory treatment options through a pathway called Expanded Access (EA). TEAMSS (Transforming Expanded Access to Maximize Support and Study) sought to examine single-patient cases to better characterize these patients. METHODS/STUDY POPULATION: We prospectively collected data on requests for single-patient EA at any one of the four collaborating TEAMSS institutions (Duke University, University of Rochester, University of Michigan, and University of Texas Southwestern) between September 1, 2021 and February 28, 2023. Regulatory and health records were reviewed for past cases that occurred between June 1, 2018 and August 31, 2021. Descriptive statistics were performed on data from the submission process, the patient demographics, the indication for treatment, and patient health status over time. RESULTS/ANTICIPATED RESULTS: The patient population was representative with respect to the largest racial groups (69.3% White / 13.0% Black or African American) and legal sex (51.3% male / 48.7% female). All ages were represented, with overrepresentation of those 60-70 years old (16.8%) and under 10 (14.8%). Patients were most often treated for infectious diseases (44.2%) or oncologic conditions (39.0%). Those who received more than one dose stayed on treatment for 76 days (median) and up to 1427 days (maximum). At the end of study, 53.9% had completed treatment as planned, moved to commercial product, or continued treatment. Death, disease progression, or failure to respond occurred for 31.9% of patients. DISCUSSION/SIGNIFICANCE: The population that receives Expanded access treatments is heterogeneous in both demographics and medical conditions. Some successful treatments are continued for years. Many patients complete their treatment, and a minority experience death or disease progression during treatment.

Objectives/Goals: Our main objective was to compare 5-year survival and organ function between patients with sickle cell disease (SCD) who underwent hematopoietic cell transplant (HCT) and those who did not undergo HCT. We hypothesized that organ function would be improved in those with SCD who underwent HCT when compared to those who remained on standard therapy. Methods/Study Population: This IRB-approved, retrospective study includes patients with SCD treated at Children’s Healthcare of Atlanta. Cases underwent HCT between 2010 and 2016. They were randomly matched with 2 patients with SCD who did not undergo HCT. Match criteria included age, sex, disease genotype, and disease severity, which was determined by the number of hospitalizations in the 5 years pre-HCT, prior intensive care unit admission, and prior chronic transfusion therapy. Data extracted included SCD treatment, hospitalizations, emergency department visits, and organ function pre-HCT and 1-, 2-, 3-, and 5-years post-HCT. Organ-specific outcomes and overall survival were compared between the two groups using cumulative incidence curves and Kaplan–Meier analyses. Normal FEV1 and FVC in this analysis were >80% predicted. Results/Anticipated Results: Thirty-seven cases who had undergone HCT were matched with 74 controls who continued with standard medical therapy. The median age was 8 years for both groups and 59% were females. The median disease severity score was 2 in both groups. At baseline, 70.3% of the HCT group completed pulmonary function tests (PFTs) compared to 35.1% of the non-HCT group. Of these, 73% in both groups had a normal FEV1. In terms of FVC, 57.7% of HCT patients and 76.9% of non-HCT patients had a normal FVC pre-HCT. At 5 years post-HCT, 56.8% of the HCT group had PFTs completed compared to 21.6% of the non-HCT group. Among these, 85.7% in the HCT group had a normal FEV1 compared to 75% in the non-HCT group, while 90.6% had a normal FVC in the HCT group compared to 75% in the non-HCT group. Two of 37 in the HCT group and 1 of 74 in the non-HCT group died (p = 0.21). Discussion/Significance of Impact: Our data suggest that post-HCT, the proportion of patients falling in the normal range for FEV1 and FVC increases. This increase is not seen in the non-HCT group, indicating that HCT may improve this organ function. There was no difference in survival between the groups, indicating the risk of HCT mortality may not be greater than the risk of living with SCD.

The efficient and effective movement of research into practice is acknowledged as crucial to improving population health and assuring return on investment in healthcare research. The National Center for Advancing Translational Science which sponsors Clinical and Translational Science Awards (CTSA) recognizes that dissemination and implementation (D&I) sciences have matured over the last 15 years and are central to its goals to shift academic health institutions to better align with this reality. In 2016, the CTSA Collaboration and Engagement Domain Task Force chartered a D&I Science Workgroup to explore the role of D&I sciences across the translational research spectrum. This special communication discusses the conceptual distinctions and purposes of dissemination, implementation, and translational sciences. We propose an integrated framework and provide real-world examples for articulating the role of D&I sciences within and across all of the translational research spectrum. The framework’s major proposition is that it situates D&I sciences as targeted “sub-sciences” of translational science to be used by CTSAs, and others, to identify and investigate coherent strategies for more routinely and proactively accelerating research translation. The framework highlights the importance of D&I thought leaders in extending D&I principles to all research stages.

The United States Government (USG) public-private partnership “Accelerating COVID-19 Treatment Interventions and Vaccines” (ACTIV) was launched to identify safe, effective therapeutics to treat patients with Coronavirus Disease 2019 (COVID-19) and prevent hospitalization, progression of disease, and death. Eleven original master protocols were developed by ACTIV, and thirty-seven therapeutic agents entered evaluation for treatment benefit. Challenges encountered during trial implementation led to innovations enabling initiation and enrollment of over 26,000 participants in the trials. While only two ACTIV trials continue to enroll, the recommendations here reflect information from all the trials as of May 2023. We review clinical trial implementation challenges and corresponding lessons learned to inform future therapeutic clinical trials implemented in response to a public health emergency and the conduct of complex clinical trials during “peacetime,” as well.

We used Cerner Real-World Data™, representing hospital admission records from 2020, to examine patients with co-occurring sickle cell disease and COVID-19 by discharge disposition grouped as death/hospice versus transfers to other facilities, returned home, or left against medical advice. Among the death/hospice group, we found older age and higher rates of congestive heart failure and diabetes. There were also significant differences in tachypnea, mechanical ventilation, minimum O2 saturation, and length of stay with higher rates in the death/hospice group. Awareness of such factors and associated mortality risks for this population may aid in patient care.

Women remain underrepresented in National Institutes of Health (NIH) study sections, panels of scientists who review grant applications to inform national research priorities and funding allocations. This longitudinal, retrospective study examined the representation of women on study sections before and during the COVID-19 pandemic. Overall, 16,902 reviewers served on 1,045 study sections across 2019, 2020, and 2021, of which 40.1% (n = 6,786) were women. The likelihood of reviewers being women significantly increased from 2019 to 2021, except among chairpersons. Understanding the representation of scientists influencing NIH grant decisions is important to ensuring scientific discovery that meets the nation’s pluralistic needs.

OBJECTIVES/SPECIFIC AIMS: Immune cells within the tumor microenvironment (TME) play an important role in the development and progression of non-small cell lung cancer (NSCLC). However, data evaluating the impact of individual immune cell types on NSCLC outcomes is limited and often conflicting. We performed a meta-analysis of existing data and used The Cancer Genome Atlas (TCGA) to evaluate the effect of several immune cells on surgical outcomes of stage I-IIIA NSCLC. METHODS/STUDY POPULATION: PubMed was searched to identify eligible studies evaluating survival of surgically resected stage I-IIIA NSCLC patients according to immune cell infiltration. Meta-analysis was performed using a linear mixed-effects model to determine overall, disease specific and progression free survival. We then used a similar patient subset found in the TCGA to validate the meta-analysis findings. For the TCGA analysis, sample-specific scores for different immune cells were computed via xCell using level three RNAseq data. After stratifying the cohort by histologic subtype, the association between each cell type and survival was assessed via Cox Regression, while adjusting for stage, gender and smoking status. RESULTS/ANTICIPATED RESULTS: From the meta-analysis (37 articles eligible; N = 8,162 patients), high levels of CD20+ B cells (hazard ratio [HR]: 0.36, 95% confidence interval [CI]: 0.15-0.85), natural killer (NK) cells (HR: 0.64, 95% CI: 0.41-1.0), and dendritic cells (0.34, 95% CI: 0.13-0.84) were significantly associated with better overall survival (OS); T regulatory cells (HR: 1.85, 95% CI: 1.35-2.54) were associated with worst OS. High CD8+ T cell infiltrates were associated with improved disease-free survival (DFS; HR: 0.85, 95% CI 0.73-0.99), while CD68+ macrophages (HR> 2.83, 95% CI: 1.28-6.24) were associated with worst DFS. In the TCGA cohort, lung adenocarcinomas rich in CD4 T cells, CD8 T cells, B cells, and NK cells were associated with improved OS in unadjusted analysis. In adjusted analysis, only NK cells were associated with improved OS (HR: 0.82, 95% CI: 0.69-0.98). There was no significant association of any immune cell type for DFS in lung adenocarcinomas and with both OS and DFS in Squamous Cell Lung Cancers (p>0.05 for all comparisons). DISCUSSION/SIGNIFICANCE OF IMPACT: The presence of tumor infiltration by specific immune cell subsets may potentially predict survival outcomes in resected stage I-III NSCLC patients. However, the impact of immune cells may not be similar in all histologic types and after adjusting for important clinical confounders.

Objectives/Goals: As life expectancy increases in people with HIV, neurocognitive impairment is becoming more common, and women with HIV (WWH) are disproportionately impacted. This work investigated mitochondrial function and oxidative stress in WWH in order to understand the relationship between mitochondrial function and cognition in future studies. Methods/Study Population: Peripheral blood mononuclear cells were isolated from virally suppressed WWH (n = 64) and underwent the Seahorse Cell Mito Stress test to assess different realms of mitochondrial function. Cells were then lysed for direct DNA extraction, and quantitative PCR was performed to understand mitochondrial DNA expression (mtDNA) levels as a measure of oxidative stress. A series of simple linear regressions was then conducted to understand the relationships between mitochondrial function and mtDNA content. Future work will expand this analysis to investigate associations between demographic dynamics, such as trauma history, and mitochondrial function, as well as to understand the relationships between mitochondrial function and cognitive outcomes in WWH. Results/Anticipated Results: In our cross-sectional analysis of mitochondrial dynamics in WWH, we found a significant association between maximum mitochondrial respiration ability and mtDNA content, with greater mtDNA expression associated with increased levels of maximum respiration following stimulation. There was no association between basal respiration and levels of oxidative stress. There was also a significant variation in mitochondrial function in our participants, indicating that future analyses to investigate the source of that variation are warranted. The work presented here sheds light on mitochondrial dynamics in WWH and will be the basis for future studies that will investigate how demographic dynamics may be associated with mitochondrial function, as well as how mitochondrial dynamics may predict cognitive outcomes. Discussion/Significance of Impact: There is significant variation in mitochondrial function in WWH. More analysis is needed to understand what may be associated with these variations, including an investigation of both clinical factors as well as cognitive outcomes. This analysis will inform directions for future mechanistic work aimed at mitigating adverse cognitive outcomes in WWH.

Objectives/Goals: Pancreatic ductal adenocarcinoma (PDAC) is a deadly disease with a mean survival of only 11 months even with the most advanced treatment to date. The desmoplastic microenvironment of PDA is thought to play a critical role in therapy resistance. One pathway that might be responsible for resistance to immunotherapy is the CXCR4-CXCL12 axis. Methods/Study Population: In this study, we propose to evaluate the effect of CXCR4-CXCL12 inhibition on dual checkpoint inhibition in KPC mouse model of PDAC and patient-derived explants. PDAC mouse models are made with pancreatic cancer cells driven by loss of TP53 and activation of KRAS. These models are treated with PD1 inhibitor Balstilimab and an FC-modified CTLA4 Botensilimab with or without CXCR4 inhibitor BL8040. In addition, we make explants of patient tumors along with their tumors and autologous peripheral blood mononuclear cells and this model is similarly challenged with BOT/BAL and BL8040. Using immunofluorescence and flow cytometry, we quantify and evaluate the spatial relationships between different cell populations. Most notably, we evaluate the relative abundance of CD8+ T cells in control and treated conditions. Results/Anticipated Results: We expect the inhibition of CXCR4-CXCL12 axis, along with two new potent checkpoint blockers, will lead to infiltration of CD8+ T cells in both the mouse and human PDAC models. We also expect this to translate into more tumor cell killing as demonstrated by Caspase activities and tumor shrinkage. Discussion/Significance of Impact: If our hypothesis is proven in both mouse and human PDAC models, this study will serve as a basis for a phase I/II clinical trial testing this combination of drug.