OBJECTIVES/GOALS: Although the approval of immune checkpoint inhibitors (ICIs) revolutionized the treatment of metastatic breast cancer (BC), yet about 30% remain unresponsive. Since the potency of ICIs depends on the efficient presentation of tumor-specific antigens by cancer cells, compounds which increase such presentation could increase efficacy of ICIs. METHODS/STUDY POPULATION: A library of the ester and urethane derivatives of polyether ionophore antibiotic, monensin (MON) has been synthesized. MTT cell viability assays were performed on the panel of human and mouse BC cell lines, and non-cancerous breast epithelial cells to determine IC50 values of MON and its derivatives. Selectivity Indexes were calculated to identify the most selective compounds towards cancer versus non-cancer cells. Major Histocompatibility Complex (MHC) class I and II presentation and Programmed death-ligand 1 (PD-L1) expression have been determined using flow cytometry. Proteins involved in apoptosis, autophagy and immunogenic cell death were identified through immunoblotting. At least three biological replicates have been performed for each experiment. RESULTS/ANTICIPATED RESULTS: MON and several of its derivatives shown activity in nanomolar range against MDA-MB-231 human BC cell line. MON and its most potent derivatives significantly increased MHC class I and II presentation and downregulated the expression of PD-L1 in BC cell lines. DISCUSSION/SIGNIFICANCE: Present findings will lead to the development of new therapeutic approaches that can serve as single agents or be used in combination with existing ICIs for the treatment of metastatic BC. By pushing the boundaries of our understanding and developing new therapies, this research can make an impact in improving outcomes for patients with metastatic BC.

OBJECTIVES/GOALS: Aiming to better understand the molecular pathogenesis of familial partial lipodystrophy (PL), we initiated whole-exome sequencing for our patients with PL syndromes. A novel variant of early B cell factor 2 (EBF2) was identified. Here we report the biological impact of a novel truncating EBF2 variant. METHODS/STUDY POPULATION: Using 3T3-L1 and human primary subcutaneous preadipocytes, we performed loss-of-function and gene rescue experiments. All cells were cultured in DMEM with 10% bovine calf serum (Invitrogen) at 5% CO2. After lentivirus transfection, cells were grown to confluence and then exposed to adipogenesis induction media containing dexamethasone (0.25µM), insulin (1µg/ml) and isobutyl methylxanthine (0.5 mM). Total RNA was extracted using RNeasy Mini Kit (Qiagen) and cDNA was synthesized using IScript (Bio-Rad). Real-time qPCR was performed using TaqMan probes for Pparg and Fabp4, two key adipogenesis markers. RESULTS/ANTICIPATED RESULTS: Patient was found to carry a heterozygous nonsense mutation in exon 6 of EBF2, causing the premature termination of the protein at amino acid position 165. Adipogenesis was significantly suppressed in 3T3L1 cells when endogenous Ebf2 was suppressed with siRNA and lentiviral shRNA. Adipocytes with suppressed Ebf2 expression showed marked reduction of intracellular lipid content and Pparg and Fabp4 expression (>80% reduction). With lentiviral gene transfer, EBF2 fully rescued adipogenic potential, whereas the truncated variant EBF2 did not. Of note, 3T3-L1 cells transfected with the EBF2 variant displayed impaired adipogenesis, suggesting a dominant-negative effect of the EBF2 variant on adipogenesis. We confirmed the dominant effect of the EBF2 variant in human adipocyte differentiation. DISCUSSION/SIGNIFICANCE: Our data suggest that EBF2 is indispensable for adipogenesis. The loss of function and dominant-negative effect of the truncating variant of EBF2 likely plays a pathogenic role in PL. Whole exome sequencing of PL patients and ex-vivo functional analysis help identify novel gene variants and better understand the molecular pathogenesis of PL.

OBJECTIVES/GOALS: The goal was to understand the effectiveness of a novel clinical trial educational module and a corresponding initiative designed and disseminated by the Southern California Clinical and Translational Science Institute (SC-CTSI) to increase the quality of clinical trials conducted in academia. METHODS/STUDY POPULATION: The CRCs (Clinical Research Coordinators) for the initiative are asked to complete the online training. Possible study protocols are picked to be monitored by the CRCs. The monitor is instructed to study the protocol extensively and prepare for their monitoring visit. The trained monitor from the initiative then reaches out to the CRC of the study that is to be monitored and carries out the monitoring visit. Afterwards, the monitor sends initiative personnel the monitoring report, which is evaluated to see if the monitor checked everything they should have during the visit. The PI of the study is contacted with highlights from the monitoring report and improvements that they can make. RESULTS/ANTICIPATED RESULTS: The first study monitored was a site of a large NIH-sponsored study where the consent forms were signed electronically. It was found that the monitor could not access the consent forms. Therefore, the monitor could not do source data verification. The PI of the study said that they would be raising this issue with the NIH. During the monitoring visit of the second study chosen for the initiative, patient binders were specifically examined for informed consent and source documentation completeness. The charts of patients were also reviewed. The only deviation found was a missing signature in the Investigator Site File. For the last two studies, data will be reported. DISCUSSION/SIGNIFICANCE OF IMPACT: Monitors were not only able to monitor efficiently, but also able to point out deficiencies in the monitoring practices of large studies. This model could be expanded to other academic institutions to establish quality management systems to ensure data integrity and subject protection.

Objectives/Goals: We aim to enhance risk prediction in kidney transplantation outcomes by improving models of peptide antigen presentation of mismatched HLA molecules. HLA-derived peptides presented by HLA Class II to T-cells can activate an immune response, ultimately leading to graft failure. We aim to improve peptide prediction by modeling antigen processing. Methods/Study Population: T-cell epitope models for HLA mismatching struggle to predict which peptides are presented because antigen processing by proteases is not well modeled. We model antigen processing of HLA Class II proteins using 3D HLA structures (crystallography data) to create an HLA-specific antigen processing likelihood (APL) model. APL uses conformational stability measurements such as b-factor, COREX, solvent accessible surface area, and sequence entropy to predict cleavage sites from proteolysis. We will integrate APL into a T-cell epitope prediction tool for HLA-derived peptides based on donor and recipient HLA genotypes. Finally, we will associate the risk of graft failure with counts of these peptides derived from APL-integrated prediction models using a historical kidney transplant cohort from 2000 to 2023. Results/Anticipated Results: We expect that applying APL could reduce false-positive peptide binders influencing risk prediction scores. We anticipate improved peptide prediction accuracy compared to existing tools such as NetMHCIIPan, which assumes all possible peptides are equally likely to emerge from antigen processing. NetMHCIIPan is currently used by PIRCHE-II HLA mismatch risk algorithm. We expect that merging antigen processing (APL) and peptide-binding (NetMHCIIPan) models into a unified model would enhance risk stratification for graft failure. Current risk stratification still leads to poor outcomes post-transplant, especially for minority population groups. Our model can identify an alternative pool of well-matched donors and has the potential to improve equity for non-White minority candidates. Discussion/Significance of Impact: Improving the understanding of how HLA matching contributes to kidney transplant outcomes can better stratify risks for kidney transplant recipients, enable personalized treatment, and ultimately improve outcomes for those undergoing kidney transplantation to treat renal diseases.

Organizations supporting translational research and translational science, including Clinical and Translational Science Award (CTSA) hubs, provide a diverse and often changing array of resources, support, and services to a myriad of researchers and research efforts. While a wide-ranging scope of programs is essential to the advancement of translational research and science, it also complicates a systematic and unified process for tracking activities, studying research processes, and examining impact. To overcome these challenges, the Duke University School of Medicine’s CTSA hub created a data platform, Translational Research Accomplishment Cataloguer (TRACER), that provides capacity to enhance strategic decision-making, impact assessment, and equitable resource distribution. This article reviews TRACER development processes, provides an overview of the TRACER platform, addresses challenges in the development process, and describes avenues for addressing or overcoming these challenges. TRACER development allowed our hub to conceptually identify key processes and goals within programs and linkages between programs, and it sets the stage for advancing evidence-based improvement across our hub. This platform development provides key insight into facilitators that can inform other initiatives seeking to collect and align organizational data for strategic decision-making and impact assessment. TRACER or similar platforms are additionally well positioned to advance the study of translational science.

ABSTRACT IMPACT: This project will use human neuron models and bioinformatics techniques to elucidate mechanisms of cocaine neurotoxicity, which will allow treatments to be developed for minimizing or preventing neurological damage caused by cocaine abuse and overdose. OBJECTIVES/GOALS: The goals of this project are to identify genes and gene networks altered by cocaine exposure in neurons (short term), and to use these pathways to understand mechanisms of cocaine neurotoxicity for the establishment of therapeutic targets (long term). METHODS/STUDY POPULATION: To study the molecular effects of cocaine, we generated preliminary proteomics and next-generation RNA sequencing (RNAseq) data from human postmortem prefrontal cortex (Broadmann area 9 or BA9) of 12 cocaine overdose subjects and 17 controls. Future directions for this project include RNAseq analysis of neuronal nuclei sorted from human postmortem BA9 and a human induced pluripotent stem cell-derived neuron (hiPSN) model of cocaine exposure from the same postmortem subjects from whom we have brain samples. RESULTS/ANTICIPATED RESULTS: We found alterations in neuronal synaptic protein levels and gene expression, including the serotonin transporter SLC6A4, and synaptic proteins SNAP25, SYN2, SYNGR3. Pathway analysis of our results revealed alterations in specific pathways involved with neuronal function including voltage-gated calcium channels, and GABA receptor signaling. In the future, we expect to see an enhancement in neuron-specific gene expression signatures in our sorted neuronal nuclei and our hiPSN model of cocaine exposure. The hiPSN model will help elucidate which effects are due to acute versus chronic exposure of cocaine. DISCUSSION/SIGNIFICANCE OF FINDINGS: Transcriptomic signatures found with this analysis can help us understand mechanisms of cocaine neurotoxicity in human neurons. With this work and future proposed studies, we can discover targetable molecular pathways to develop drugs that can reduce or reverse cocaine-related impairment.

Objectives/Goals: Our research group is focused on care of hospitalized persons with opioid use disorder (OUD) in the era of high-potency synthetic opioids (HPSO). In this work, we describe trends in patient-directed discharge (PDD) and inpatient treatment with medications for opioid use disorder (MOUD). We hypothesized that PDD is associated with MOUD dose and timing. Methods/Study Population: Patient data generated in the routine care of patients was automatically abstracted using a SQL query on Epic Clarity tables in the electronic health record (EHR). We included adult patients admitted to Johns Hopkins Hospital between July 1, 2019 and June 30, 2022, with an ICD-10-CM code for a list of opioid-related disorders (F11.X) consistent with OUD. Demographics, prior medication list, clinical care including hospital service, consultation services, COWS scores, length of time in emergency department, time of triage, time until receipt of methadone or buprenorphine, dosage and timing of MOUD, opioid medications other than methadone or buprenorphine, adjuvant medications; prior methadone or buprenorphine treatment and disposition. Query results were validated by manual abstraction of EHR. Results/Anticipated Results: The SQL identification of the cohort of patients with OUD was found to be accurate. Time of triage, discrete orders completed during hospitalization were well represented in the query. The query was able to identify individual opioid medications but unable to summarize total dose in Morphine Milligram Equivalents. The query did not extract accurate information from patient-controlled analgesia pumps due to the continuous nature of the medication rather than discrete doses reflected in the medication administration record. Finally, the query characterized prior treatment with methadone or buprenorphine as a binary variable – dosage and timing of that prior treatment could not be accurately represented. Finally, stimulant use is not reliably collected in the EHR and was unavailable. Discussion/Significance of Impact: Given the rise of HPSO, patients may not tolerate delay of MOUD. Improving the granularity of data collected will offer more insight into the inpatient treatment for OUD. Real-world data have promise but requires extensive technical expertise. Future work is needed to improve capture of derived variables such as total dosage of opioids in MME.

OBJECTIVES/SPECIFIC AIMS: Previous studies suggest that genetic variants in the oxytocin receptor (OXTR) may alter oxytocin dose requirement for labor induction and may increase risk for preterm labor and neurodevelopmental disorders. However, the mechanisms of actions of these variants remain unknown. The goal of this study was to functionally characterize common missense and noncoding variants in OXTR. First, we aimed to determine the effects of missense variants on two major aspects of receptor function: calcium signaling and β-arrestin recruitment. Second, we used allelic expression imbalance assays in an effort to identify regulatory single nucleotide polymorphisms (SNPs) in noncoding regions of OXTR that alter OXTR mRNA expression. METHODS/STUDY POPULATION: We used the Exome Aggregation Consortium database to identify the 12 most prevalent missense single nucleotide variants in OXTR. To determine the functional effects of these variants, we transfected human embryonic kidney cells (a common model system used to study receptor function) with wild type OXTR, variant OXTR, or empty vector control. We used the calcium-sensitive dye Fluo4 to quantify intracellular calcium flux in response to oxytocin treatment, and used bioluminescence resonance energy transfer assays to measure recruitment of the signaling partner β-arrestin to the receptor. To investigate potential effects of noncoding SNPs on OXTR mRNA expression, we quantified allele-specific expression of OXTR in human uterine tissue obtained from participants at the time of Cesarean section. We used next-generation sequencing (Illumina MiSeq) to count alleles of a reporter SNP in OXTR exon 3. RESULTS/ANTICIPATED RESULTS: Of the 12 most prevalent missense single nucleotide variants, four were predicted to be deleterious by PolyPhen variant annotation software. We anticipate that these variants will alter receptor signaling through calcium or β-arrestin pathways. We further observed that a reporter SNP in OXTR exon 3 exhibits significant allelic expression imbalance in a subset of our myometrial tissue samples, indicating that OXTR expression may be regulated by a functional SNP. Our current work focuses on discovering the functional SNPs in OXTR responsible for the pattern of allelic expression imbalance seen in mRNA. In the future, we will seek to explore the effects of these variants on uterine function by using genome editing of uterine smooth muscle cells. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results suggest that both missense and noncoding variants may affect OXTR expression and function. Future studies may suggest that OXTR sequencing, genotyping, or expression analysis would be useful to identify individuals likely to respond or fail to respond to safe doses of oxytocin for labor induction. Personalizing approaches for labor induction in this way would increase the safety of oxytocin and potentially reduce maternal morbidity and mortality.

OBJECTIVES/GOALS: Clinical research is the backbone of the medical community. However, there are few regulations to ensure clinical trial participants can understand their results, leading to volunteers feeling unvalued and unlikely to enroll in trials1. This study examines the need of lay summaries METHODS/STUDY POPULATION: To understand the current landscape of clinical trial summaries, literature searches were conducted using the University of Southern California Library database with keywords Title contains “lay language” OR “lay summary” AND any field contains “Trial” OR “clinical”, and Title contains “natural language processing” AND “clinical trial” OR “Summary”. Studies were deemed relevant if they discussed lay language summaries for health care realms or using Natural Language Processing (NLP) to increase comprehension. Papers published by the Center for Information and Study on Clinical Research Participation (CISCRP) were reviewed and their Associate Director was interviewed. RESULTS/ANTICIPATED RESULTS: Of 67 total results, 14 were determined to be relevant. Ten of the relevant results examined lay language summaries and their regulation and 4 were NLP studies. The European Medicines Agency set regulations mandating clinical trial summaries. However, researchers have difficulty validating to an appropriate reading level2. Difficulty and potential bias halted a U.S. mandate of lay summaries3. The nonprofit CISCRP has partnered with industry to develop unbiased clinical trial summaries resulting in all volunteers feeling appreciated and 91% understanding clinical trial results post summary1. Similarly, NLP software for annotating Electronic Health Records increased comprehension for 77% of patients4. DISCUSSION/SIGNIFICANCE OF IMPACT: In the U.S., a lack of regulations mandating lay summaries may be related to concerns by regulatory agencies that summaries in plain language may introduce bias3. Future looks into integration of NLP systems to clinical trials may create unbiased summaries and allow for FDA regulation.

OBJECTIVES/GOALS: Navigating the NIH Single IRB Policy has been challenging for investigators, study teams, and Human Research Protection Programs (HRPP). In response, the Indiana Clinical and Translational Sciences Institute (CTSI) created an innovative Single IRB Project Manager role (sIRB PM), uniquely placed within the Indiana CTSI. METHODS/STUDY POPULATION: The Single IRB Project Manager role was created in 2018 by the Indiana CTSI in response to the NIH Single IRB Policy for Multi-Site Research. The role of the sIRB PM is to serve as a liaison between the Indiana University HRPP, lead site, coordinating center, and participating sites when Indiana University serves as the Single IRB. This model has proven useful to both the IRB and lead site, notably in the following ways:

• At study start-up, the sIRB PM can handle complicated communications among sites and the IRB at the same time the lead site is responsible for many other administrative tasks related to start-up. By absorbing the workload of IRB approval for multiple sites, the sIRB PM provides the lead site more capacity to handle other essential tasks.

• The sIRB PM translates new terminology and facilitates processes that are new for sites.

RESULTS/ANTICIPATED RESULTS: Early assessment of this program is predominantly positive. The sIRB PM currently supports 24 external sites. In an NIA-funded 13 site study, all sites were added within 9 months of initial IRB approval of the protocol. This role fills a gap that benefits:

• IRB staff by allowing them to fulfill their duties of screening and review while leaving some of the reliance organization to the sIRBPM.

• Lead PI by allowing them to focus on conducting the research instead of the many administrative tasks required for single IRB review.

• Participating sites by having a liaison to enter their amendments and reportable events into an otherwise closed IRB software system.

• All parties by having the sIRB PM manage document organization, storage, and distribution study-wide.

DISCUSSION/SIGNIFICANCE OF IMPACT: The CTSI sIRB PM role effectively shifts administrative work caused by the sIRB mandate by merging research coordinator experience with regulatory experience while building upon an existing strong relationship with the HRPP. Future focus is on process education, standardizing pricing structure, and ensuring sufficient budget support in grants.

Objectives/Goals: The research aims to prototype a mobile app for physicians to remotely monitor patients receiving CD19-directed CAR T-cell therapy post-discharge. This app will facilitate standardized data collection across carious CAR-T treatment centers and help harmonize follow-up protocols. Methods/Study Population: A literature review and semi-structured interviews with patients, clinical coordinators, and experts helped identify essential parameters for a mobile app prototype aimed at monitoring adverse effects such as cytokine release syndrome and neurotoxicity. The app was designed through process mapping to combine data from self-reports and wearable devices, such as the Garmin smartwatch. New screens were designed in Figma, drawing from an existing patient monitoring app for allogeneic stem cell transplant follow-up. Finally, a preliminary feasibility study will be conducted to gather feedback on the app prototype from CAR T-cell therapy patients, healthcare providers, and stakeholders, ensuring its effectiveness and usability. Results/Anticipated Results: Semi-structured interviews with people with professional and lived experience with CAR T-cell therapy were conducted to determine what metrics might be monitored and how they could be measured remotely to effectively monitor for side effects. The mobile phone application was then prototyped using process mapping in Visio®, designed in Figma, and preliminary development was completed. The final prototype includes parameters that will be recorded or measured using a combination of self-reporting and devices to monitor body temperature, basic vitals, activity, sleep, and cognitive function, among others. The prototype of the remote monitoring app is the first step in implementing remote monitoring of CAR T patients, standardized data collection, and reduction in the overall cost of CAR T-cell therapy. Discussion/Significance of Impact: This app will enable physicians to monitor patients for routine follow-ups and adverse effects, such as CRS and ICANS. Future research will validate the digitized ICANS assessment and used to establish best practices for standardizing CAR-T follow-up protocols across Canada.

OBJECTIVES/GOALS: Maternal health and exposures during pregnancy play a major role in shaping the neurodevelopment of our offspring—one influence is maternal immune activation (MIA). Here we explore the association of MIA during pregnancy and the developing human connectome through analysis of 46 markers of activation. METHODS/STUDY POPULATION: 74 healthy women with singleton pregnancies underwent blood draws between 34-37 weeks gestation. 46 markers of maternal immune activation, both adaptive (e.g., IgG) and innate (e.g., cytokines and acute phase reactants), were collected. In addition, for preliminary analyses of MIA in relation to the newborn brain, we utilized 30 participants with MRIs between the ages of 0-6 months. RESULTS/ANTICIPATED RESULTS: Principal component analysis (PCA) identified the first 5 PCs explains ~68% of the variance and the first 10 explains ~83% (top PC is 42.1%). Using the top PC each edge in the connectome was correlated with the immune profiles. Several regions trended towards significance–one survived correction and included 359 edges, showing. The highest number of edges was observed in the inferior parietal lobe of the left hemisphere–a region associated with functions from basic attention toa social cognition, suggesting that deviations in fetal exposure to MIA can longitudinally impact offspring behavior in areas essential for human interaction. DISCUSSION/SIGNIFICANCE: This is the first study in understanding how interruptions (i.e., MIA) influence later development. Identification of alterations, and long-term outcomes could lead to the development of mechanism-based healthcare, facilitate timely referral for appropriate interventions and provide family support.

OBJECTIVES/GOALS: RETgene fusions in sarcoma are rare and their impact on pathogenicity is unknown. Malignant peripheral nerve sheath tumors (MPNST) are a deadly, genomically heterogenous soft tissue sarcoma rarely harboring targetable aberrations. We present a case of a CCDC6-RET fusion MPNST sensitive to RET-inhibitor therapy in a xenograft model. METHODS/STUDY POPULATION: Lung tumor tissue was obtained per an approved collection protocol from a 21yo male patient with a spontaneous MPNST harboring an inactivating mutation in NF-1 and a CCDC6-RET gene fusion detected by a commercially available sequencing panel (Signatera). To confirm pathogenicity of the RET fusion, fresh tumor tissue was engrafted into immunocompromised NSG mice in the anterior and posterior flanks, harvested at ~10 weeks, and re-transplanted into bilateral flanks. When tumor diameters reached 0.5-1cm (~4 weeks), mice were randomized into 3 groups (n=6/group) and treated with either vehicle (V) (PBS), the RET-specific inhibitor selpercatinib (S) (20mg/kg twice daily), or the multi-kinase inhibitor cabozantinib (C) (30mg/kg daily) by oral gavage. Mice were monitored weekly for weight and tumor size. RESULTS/ANTICIPATED RESULTS: 92% (33/36) of implanted tumors were evaluable for treatment response. Pre-treatment tumor volumes (mm3) across all three groups were similar (mean/Std Dev – V: 230/111, S: 271/132, C: 230/123). At day 7, tumor growth was significantly inhibited by S and C versus V (ANOVA p < 0.001, post-hoc Tukey’s V vs S p= 0.0178, V vs C p< 0.0001, S vs C p= 0.0005). V-treated tumors increased in volume by 60% while S reduced tumor volume by ~80% and C reduced tumor volume by ~20%. S and C treatments were tolerated well. and S improved survival with 100% of mice alive at day 63 vs 0% in V and C groups. 6 of the 12 implanted tumors treated with S, 50% increased in size after ~6-weeks following a >90% initial tumor reduction in tumor volume. Follow-on molecular studies in S-resistant tumors are ongoing. DISCUSSION/SIGNIFICANCE: Targetable genomic changes in MPNST, especially in RET, are infrequent and often considered stochastic. Our findings suggest that precision medicine approaches pairing genomic sequencing and in vivotesting of target gene pathogenicity may guide treatment planning and novel discovery for rare, difficult to treat sarcomas.

OBJECTIVES/GOALS: Distinguishing indolent from aggressive prostate cancer and early identification of men at risk of developing aggressive, metastatic disease is of great importance. We aim to explore the relationship between N-glycan and collagen composition in prostate tumor tissue and the long-term outcome of the disease. METHODS/STUDY POPULATION: Matrix assisted laser desorption/ionization mass spectrometry can be utilized to characterize N-glycan profiles in formalin fixed paraffin embedded tissues. Collagen may also be characterized using ECM-targeted collagenase MALDI imaging. These approaches were used to analyze prostatectomy samples with different clinical outcomes. Tissue microarrays containing tissues from 75 non-progressors (no evidence of disease; NED) and 50 metastatic cases (MET) were examined. From a combined list of 90 N-glycans and 500 collagenase peptides, the average AUC intensity value for each glycan and collagen peptide was extracted and assessed as a predictor of metastatic progression. Ancestral informative markers were analyzed and polygenic hazard risk scores were generated for samples as well. RESULTS/ANTICIPATED RESULTS: Three N-glycans and three collagen peptides were found to discriminate between NED and MET cases with statistical significance. The best performing N-glycan was Hex6HexNAc6Fuc1 with an AUC of 0.77 (p<0.001). While the best performing collagen peptide was COL1A2 with an AUC of C 0.77 (p<0.001). DISCUSSION/SIGNIFICANCE: Both a collagen peptide and N-glycan were discovered as promising biomarkers to predict metastasis. Future validation studies are needed to confirm biomarker potential and to determine if the addition of these biomarkers can strengthen current genomic classifier’s ability to predict metastatic prostate cancer.

OBJECTIVES/GOALS: Accurate classification of disorders of consciousness (DoC) is key in developing rehabilitation plans following brain injury. The Coma Recovery Scale-Revised (CRS-R) is a sensitive measure of consciousness. We explore feasibility, safety and impact of CRS-R guided rehab in hemorrhagic stroke patients with DoC and evaluate predictors of recovery. METHODS/STUDY POPULATION: Consecutive patients with non-traumatic hemorrhagic stroke, defined as subarachnoid hemorrhage (SAH) or intracerebral hemorrhage (ICH), receiving serial CRS-R assessments during their ICU stay at University of Maryland Medical Center from 2017-2021 were retrospectively identified. Outcomes of interest included the association with CRS-R and discharge disposition, therapy-based function and mobility and occurrence of safety events during CRS assessment. We also examined the association between CRS-R and physiological and anatomical injury pattern on electroencephalography (EEG) and magnetic resonance imaging (MRI), respectively. CRS-R RESULTS/ANTICIPATED RESULTS: 76 patients with≥2 CRS-R assessments were identified (22 SAH, 54 ICH, median age = 59, 50% female). Median CRS-R completed was 3 with no SAEs identified during sessions. We identified 4 patterns: persistent VS/UWS (49%), persistent MCS or better (13%), emergence from VS/UWS to MCS or better (27%) and regression from MCS or better to VS/UWS (11%). Persistent low CRS-R correlated with older age in SAH (p=0.01), female gender in ICH (p=0.04), and history of diabetes (p=0.01). 2% of patients with final CRS-R DISCUSSION/SIGNIFICANCE: Early neurorehabilitation guided by CRS-R appears to be feasible and safe acutely following hemorrhagic stroke complicated by prolonged DoC and may enhance access to inpatient rehabilitation with a lasting benefit on recovery. Further characterization of DoC patterns and their correlation to clinical markers, including EEG and MRI is needed.

ABSTRACT IMPACT: Chronic kidney disease (CKD) affects ˜15% of the US population and the majority of patients are diagnosed too late to benefit from early intervention. We are developing a new diagnostic imaging tool (RadioCF-PET) for the kidney to enable early detection of diseases and to monitor overall kidney health. OBJECTIVES/GOALS: Nephron mass, or the number of functioning nephrons, is a measure of the functional capacity of the kidney. RadioCF-PET may enable early detection of nephron loss in patients with or at risk of CKD before changes are clinically detectable, facilitating early interventions to improve outcomes in these patients. METHODS/STUDY POPULATION: RadioCF-PET, labeled with Cu-64, has the advantage of using sub pharmacological doses for imaging, carrying low risk and can be used with the FDA’s exploratory IND (eIND) mechanism for early in human testing. We are developing the technology to be used in pre-eIND toxicology and pharmacology studies. We are also developing other aspects of translational science to propel this technology toward translation, including: market analysis, critical path to market, customer discovery, and commercialization strategy. RESULTS/ANTICIPATED RESULTS: Milestone 1: Apply technology in mouse model study and in human kidneys rejected for transplant. Anticipated Result 1: PET signal from RadioCF-PET correlates with glomerular density in healthy and diseased model male mice (R2 = 0.98). RadioCF-PET signal correlates with glomerular number in a donated human kidney (R2= 0.78). Milestone 2: Application to federal funding (STTR) and gap funding mechanisms to enable pre-eIND studies. Anticipated Result 1: Application for funding will aid to clarify and validate our market analysis and commercialization strategy. Milestone 3: Continued research and development with the technology in new studies. Other Anticipated Results: Future work with RadioCF-PET will enhance technology performance in preparation for pre-eIND studies. DISCUSSION/SIGNIFICANCE OF FINDINGS: We foresee a large clinical and commercial potential for RadioCF-PET to provide precise, early monitoring in patients at risk for or with CKD. The two biggest hurdles for clinical translation are validating safety and proving efficacy. This work targets both issues to facilitate RadioCF-PET toward clinical translation.