The last 50 years have seen an increasing dependence on academic institutions to develop and commercialize new biomedical innovations, a responsibility for which many universities are ill-equipped. To address this need, we created LEAP, an asset development and gap fund program at Washington University in St. Louis (WUSTL). Beyond awarding funds to promising projects, this program aimed to promote a culture of academic entrepreneurship, and thus improve WUSTL technology transfer, by providing university inventors with individualized consulting and industry expert feedback. The purpose of this work is to document the structure of the LEAP program and evaluate its impact on the WUSTL entrepreneurial ecosystem. Our analysis utilizes program data, participant surveys, and WUSTL technology transfer office records to demonstrate that LEAP consistently attracted new investigators and that the training provided by the program was both impactful and highly valued by participants. We also show that an increase in annual WUSTL start-up formation during the years after LEAP was established and implicate the program in this increase. Taken together, our results illustrate that programs like LEAP could serve as a model for other institutions that seek to support academic entrepreneurship initiatives.

The Centers for Medicare & Medicaid Services have mandated that hospitals implement measures to screen social determinants of health (SDoH). We sought to report on available SDoH screening tools. PubMed, Scopus, Web of Science, as well as the grey literature were searched (1980 to November 2023). The included studies were US-based, written in English, and examined a screening tool to assess SDoH. Thirty studies were included in the analytic cohort. The number of questions in any given SDoH assessment tool varied considerably and ranged from 5 to 50 (mean: 16.6). A total of 19 SDoH domains were examined. Housing (n = 23, 92%) and safety/violence (n = 21, 84%) were the domains assessed most frequently. Food/nutrition (n = 17, 68%), income/financial (n = 16, 64%), transportation (n = 15, 60%), family/social support (n = 14, 56%), utilities (n = 13, 52%), and education/literacy (n = 13, 52%) were also commonly included domains in most screening tools. Eighteen studies proposed specific interventions to address SDoH. SDoH screening tools are critical to identify various social needs and vulnerabilities to help develop interventions to address patient needs. Moreover, there is marked heterogeneity of SDoH screening tools, as well as the significant variability in the SDoH domains assessed by currently available screening tools.

The ACT Network was funded by NIH to provide investigators from across the Clinical and Translational Science Award (CTSA) Consortium the ability to directly query national federated electronic health record (EHR) data for cohort discovery and feasibility assessment of multi-site studies. NIH refunded the program for expanded research application to become “Evolve to Next-Gen ACT” (ENACT). In parallel, the US Food and Drug Administration has been evaluating the use of real-world data (RWD), including EHR data, as sources of real-world evidence (RWE) for its regulatory decisions involving drug and biological products. Using insights from implementation science, six lessons learned from ACT for developing and sustaining RWD/RWE infrastructures and networks across the CTSA Consortium are presented in order to inform ENACT’s development from the outset. Lessons include intentional institutional relationship management, end-user engagement, beta-testing, and customer-driven adaptation. The ENACT team is also conducting customer discovery interviews with CTSA hub and investigators using Innovation-Corps@NCATS (I-Corps™) methodology for biomedical entrepreneurs to uncover unmet RWD needs. Possible ENACT value proposition hypotheses are presented by stage of research. Developing evidence about methods for sustaining academically derived data infrastructures and support can advance the science of translation and support our nation’s RWD/RWE research capacity.

This paper describes the development and validation of a new 32-item test of knowledge of good clinical practice (GCP) administered to 625 clinical research coordinators. GCP training is mandated by study sponsors including the US National Institutes of Health. The effectiveness of training is rarely assessed, and the lack of validated tests is an obstacle to assessment. The GCP knowledge test was developed following evaluation of two existing widely used GCP tests to ensure it accurately reflects the content of current training. The final GCP knowledge test demonstrated good reliability (α = 0.69). It is a valid and reliable instrument for measuring knowledge of GCP. The test will be useful in assessing the effectiveness of GCP training programs as well as individuals’ mastery of GCP content.

Clinical trial participation among historically underrepresented populations remains low in large part due to mistrust of academic institutions and research investigators. Mistrust may be ever greater today given misinformation related to COVID-19. The Research Ambassador Program is an interactive educational workshop delivered by Promotoras de Salud/Community Health Workers and designed to both address common myths, fears, and concerns about research and encourage research participation among underrepresented populations. An evaluation conducted with 819 Latino and Black participants demonstrated a change in behavior and intention to participate in a clinical trial, with half of participants enrolling in a clinical trial research registry.

ABSTRACT IMPACT: This research takes a transcriptomic approach to parse genes and molecular pathways that underlie the fear memory circuitry and, in doing so, identifies therapeutic targets that can further be developed into treatments for fear disorders, such as post-traumatic stress disorder. OBJECTIVES/GOALS: Normal fear learning produces avoidance behavior that promotes survival, but excessive and persistent fear after trauma can lead to development of phobias and post-traumatic stress disorder (PTSD). Our goal is to understand the mechanism and identify novel genetic targets underlying fear responses. METHODS/STUDY POPULATION: Involvement of the basolateral amygdala (BLA) in fear acquisition is well established and requires activation of N-methyl-D-aspartic acid receptors (NMDARs). At a cellular level, NMDAR activation leads to production of nitric oxide (NO) by a process mediated by interaction between postsynaptic density protein 95 (PSD95) and neuronal nitric oxide synthase (nNOS). To elucidate mechanisms underlying the role of the PSD95-nNOS-NO pathway in conditioned fear, here we use rodent behavioral paradigms, pharmacological treatment with a small molecule PSD95-nNOS inhibitor, RNA-sequencing, and an AAV-mediated knockdown of the nNOS gene in the BLA. RESULTS/ANTICIPATED RESULTS: We show that treatment with ZL006 attenuates rodent cued-fear consolidation. Additionally, with RNA-sequencing, expression of 516 genes was altered in the BLA following fear expression; of these genes, 83 were restored with systemic ZL006 treatment. Network data and gene ontology enrichment analyses further revealed that cGMP effects, insulin-like growth factor binding, and cognition-related pathways were significantly altered. Finally, we show that a BLA-specific knockdown of nNOS attenuates cued fear consolidation, without adverse effects on other memory and motor behaviors. DISCUSSION/SIGNIFICANCE OF FINDINGS: Via a model of NMDAR-mediated fear consolidation, our results reveal novel pathways and genetic targets that underlie plasticity of fear memory circuitry. Importantly, these results will inform future therapeutic strategies for targeting fear related disorders like PTSD.

OBJECTIVES/GOALS: In the United States, oropharyngeal cancer (OPC) is the leading human papilloma-virus (HPV) related malignancy, and OPC rates are increasing among the US veteran population. The purpose of this study is to identify demographic and regional factors that may be associated with low HPV-vaccination rates among the US veteran population. METHODS/STUDY POPULATION: This study will use Veterans Health Administration (VHA) administrative data to create a national cross-sectional cohort of veterans ages 18-45 with a VHA primary care visit from 2018-2020. HPV-vaccination status of each subject (initiation of vaccination series, completion of vaccination regimen, and age vaccinated) will be determined. Differences in the prevalence of HPV-vaccination by smoking status, geographic location, socioeconomic status, race/ethnicity, and rural-urban context will be examined in the unadjusted analysis. Factors associated with low HPV-vaccination rates in the VHA will be identified using multivariable logistic regression to model no (vs any) HPV-vaccination, no completion (vs completion) of HPV-vaccination recommendations, and non-routine (vs routine) HPV-vaccination: RESULTS/ANTICIPATED RESULTS: In 2019, HPV-vaccination guidelines were expanded to include some adults between the ages of 26-45, making many young veterans in the VHA eligible for vaccination. From this study’s recently generated dataset, more than 1.2 million subjects (n=1,219,896) met the study inclusion criteria. Extrapolating from trends in the civilian population, it is anticipated that HPV-vaccination rates will be lower among African Americans compared to non-Hispanic Caucasian Americans, within the South Central and Southeastern regions of the US, and in rural communities. This study will define a veteran’s geographic location by their associated Veteran Affairs Integrated Service Network (or VISN), which are groups responsible for healthcare planning and resource allocation in particular regions of the US: DISCUSSION/SIGNIFICANCE: Identifying factors associated with low HPV-vaccination rates within the VHA will be the first step to reducing future incidence of HPV-related cancer burden among US veterans. Assessing geographic variability in vaccination rates by VISNs will create the potential to generate targeted interventions within an existing VHA framework.

OBJECTIVES/SPECIFIC AIMS: Infants with single ventricle congenital heart disease (CHD) who undergo staged surgical reconstruction are among the pediatric patients at highest risk for thrombotic complications. Despite improvements in survival due to medical and surgical advancements, thrombotic complications are common and lead to increased morbidity and mortality, especially during the first two stages of surgical reconstruction. The burden of disease caused by thrombosis is not fully known, and the risk factors associated with thrombosis are not clear. Due to this knowledge gap, prevention of thrombosis with medication, a strategy called thromboprophylaxis, has not been standardized, leading to inadequate prevention of thrombosis. In order to understand the burden of thrombosis and then provide targeted thromboprophylaxis for thrombosis prevention, better characterization of thrombotic complications and the associated factors is needed. Hypothesis: I hypothesize that in infants with single ventricle CHD, the incidence of thrombosis will be more frequent after stage I Versus stage II reconstruction, despite the type of shunt used. Specific demographic, clinical, and surgical variables will be associated with an increased risk for thrombotic complications, and a model to predict which subset of infants is at increased risk will be developed. Specific Aim 1: Characterize the incidence of thrombotic complications at different time points from stage I through stage II of the single ventricle reconstruction (SVR) pathway and determine the demographic, clinical, and surgical factors associated with thrombosis in infants with single ventricle CHD. (1) Determine the incidence of thrombosis in infants with single ventricle CHD. (2) Compare the rate of thrombotic complications between the 2 most commonly used approaches for stage I reconstruction for the group of patients with hypoplastic left heart type of anatomy [modified Blalock-Taussig shunt (MBTS) vs. right ventricle to pulmonary artery shunt (RVPAS)]. (3) Determine the factors (demographic, clinical, and surgical) associated with thrombosis in infants with single ventricle CHD. Specific Aim 2: Determine which subset of infants with single ventricle CHD is at increased risk of developing thrombotic complications across the first 2 stages of surgical reconstruction. (1) Test the identified demographic, clinical, and surgical variables including, but not limited to, gestational age, sex, CHD diagnosis, baseline oxygen saturation, stage of reconstruction, shunt type, and other clinical data available in a univariable and multivariable analysis and study their potential interactions to construct a novel risk predictive model specific for single ventricle CHD. METHODS/STUDY POPULATION: To address the specific aims, I will utilize data from the SVR clinical trial public use data set. This data set includes a prospective cohort of infants, 0–14 months of age, enrolled from any of the 15 participating clinical centers from the years 2005 to 2009. Inclusion criteria for enrollment were diagnosis of hypoplastic left heart syndrome or related single, morphologic right systemic ventricle anomaly, planned Norwood procedure, and informed consent of parent or legal guardian. No additional subjects outside of this data set will be included. Exclusion criteria were a diagnosis of single, morphologic left ventricle anomaly, preoperative identification of anatomy rendering the MBTS or RVPAS technically impossible, and any other major abnormality or acquired extra-cardiac disorder that could independently affect the likelihood of the subject meeting the primary endpoint. The complication of stroke will be excluded from the analyses of factors associated with thrombosis. The complication of thrombosis as defined in this dataset is a composite of events that include arterial or venous thrombosis, thromboembolism, and pulmonary embolism. The data was collected in such a way that it will not be possible to separate these sub-types of thrombosis. Additional thrombotic events of interest are superior vena cava occlusion and inferior vena cava occlusion. Specific Aim 1: Patient data will be extracted from the SVR clinical trial public use dataset to characterize the incidence of thrombotic complications at different time points from stage I through stage II of the SVR pathway and determine the demographic, clinical, and surgical factors associated with thrombosis in infants with single ventricle CHD. In addition, I will compare the rates of thrombotic complications between the 2 most commonly used approaches for stage I palliation for the group of patients with hypoplastic left heart type of anatomy (MBTS vs. RVPAS) and will test the hypothesis that the risk of thrombotic complications is associated with the stage of palliative surgery (stage I vs. stage II). Specific Aim 2: We will test identified demographic, clinical, surgical, and newly identified variables in a univariable and multivariable analysis and study their potential interactions to construct a novel risk predictive model specific for single ventricle CHD. RESULTS/ANTICIPATED RESULTS: To determine feasibility for adequate numbers to be able to address the research aims, a preliminary analysis dataset was performed using a dataset from the Pediatric Heart Network. The PHN is a collaborative group of hospitals that participates in clinical research studies in children with CHD. For the SVR clinical trial, the PHN conducted a randomized clinical trial at 15 centers in North America between 2005 and 2009, prospectively enrolling infants with HLHS or single right ventricle anomalies who were to undergo the Stage I Norwood procedure. A total of 920 newborns were screened; 664 were medically eligible and 549 patients were randomized. The primary aim of the trial was to compare survival of infants randomized to receive either the Norwood procedure with the MBTS or the RVPAS. These patients were followed at specific time points, including from baseline (pre-Norwood), at the time of the Norwood procedure, between stage I and II, following stage II reconstruction, and at 14 months of age. At these time points, data were collected that includes demographic, radiologic, clinical, and surgical outcomes. Included in the clinical outcomes are complications, such a thrombosis. There was no screening process to assess for asymptomatic thromboses, suggesting that most, if not all, discovered thromboses were due to clinically relevant effects. A newer iteration of this study (SVRIII) expands the monitoring of this cohort until the Fontan stage at 2–6 years of age, but these data have not yet been released in the public use data set. A descriptive analysis of the frequency of thrombotic complications was assessed at each time point, as well as in aggregate. Data were extracted from the specific time periods of interest, identified as Pre-Norwood, during Norwood Hospitalization, in-between visits, and during Stage II Hospitalization. There were 549 infants who were randomized with available data to analyze. During the Norwood hospitalization, 37 infants had a thrombotic complication. Between Stage I and Stage II outpatient visits, 8 infants had a thrombotic complication. During Stage II hospitalization, 16 infants had a thrombus. Overall, 61 individual patients (11%) had a thrombotic complication. DISCUSSION/SIGNIFICANCE OF IMPACT: This study utilizing data from the Pediatric Heart will be the largest cohort ever utilized for characterizing thrombotic complications and determining the factors associated with thrombosis across the first and second stages of surgical reconstruction. More than 500 (n=549) subject’s data will be analyzed through the first two stages of reconstruction, while the largest analysis before this proposed analysis only included a total of 195 children. Notably, these prior studies did not include a comparison between the 2 shunt types in stage I reconstruction, leaving a gap in knowledge regarding the incidence of thrombosis comparing these groups. The analysis will be the first to address this gap and update the current literature. Preliminary data show that the overall incidence of thrombosis across the first 2 stages of surgical reconstruction was 11%, which is lower than the previously reported overall rates of 40%–50%. Despite the continued lack of evidence-based guidelines for thromboprophylaxis methods, the decreased overall rate is most likely due to more widespread practice of anticoagulation in general. Determining the factors associated with thrombosis across the first and second stages of surgical reconstruction will help identify those at risk. An innovative aspect of this analysis will be the use of disease-specific factors to develop a model to predict thrombosis. Unique factors include cardiac variables like ejection fraction, baseline oxygen saturation, shunt type (MBTS vs. RVPAS), and other echocardiographic parameters. While the use of thromboprophylaxis has been associated with decreased risk of thrombosis, there is no general consensus to guide thromboprophylaxis in this population, which can be burdensome and costly. Determining which subset of infants with single ventricle CHD are at increased risk of developing thrombotic complications will allow for the development of a prediction model to predict those at highest risk of developing a thrombotic complication. Developing a predictive model will be a novel way to identify patients at risk for thrombosis and will set the stage for targeted prevention of thrombosis.

ABSTRACT IMPACT: Our research would be the first therapeutic to both prevent and treat osteoarthritis - helping 27 millions U.S. citizens alone immediately. OBJECTIVES/GOALS: Our objective is to conjugate hyaluronic acid binding peptides (HABP) to anionic hollow nanoparticle (hNP), and allowing the HABP-hNP complex to penetrate into osteoarthritic cartilage, bind to exposed HA, prevent further degradation, and restore the compressive strength of articular cartilage. METHODS/STUDY POPULATION: N-isopropyl acrylamide, 2-acrylamindo-2-methyl-1-propanesulfonic acid, N,N’-bis(acryoyl)cystamine, and Acrylic Acid, in fluorescent batches rhodamine b isothiocyanate (RBITC), were polymerized via precipitation reaction. HA binding peptide, GAHWQFNALTVRGSG-Hydrazide (GAH-Hyd), was covalently bonded to the hNP using DMTMM chemistry. The reaction was halted by diluting the solution 10:1 with milliQ water and purified using tangential flow filtration. The dynamic viscosity of the six treatments were analyzed in a 70 kDa HA. Using a rheometer (Discovery HR-3) with a 20 mm parallel plate geometry, TA Instruments, New Castle, DE), a frequency sweep (0.01 -1000 Hz, 2.512 Pa) was conducted to measure the storage modulus of each solution. RESULTS/ANTICIPATED RESULTS: GAH-Hyd was successfully conjugated to the surface of the hNP and zeta-potential shows a significant increase in surface charge from -21.41 mV for unconjugated hNP to -8.94 mV for 65 GAH conjugated hNP, confirming conjugation. The hNPs need 65 ±10 GAH per nanoparticle to significantly bind to HA, shown by increasing the dynamic viscosity of the solution. The minimum concentration of 65 GAH-hNP required to significantly bind to HA is 313 µM. These data from our study display the ability to functionalized the surface of polymeric hNPs with site specific peptides and their ability to bind to diseased tissue. We expect the GAH-hNP system will restore the compressive strength of OA cartilage and prevent further HA degradation in ex vivo aggrecan depleted cartilage plugs. DISCUSSION/SIGNIFICANCE OF FINDINGS: Binding to exposed HA within the ECM of cartilage protects the HA from further degradation, halting the progression of OA. 65 GAH-hNP binds to HA at a 313 µM. Our system can be translated and used to treat a multitude of conditions by conjugating tissue specific peptides to the surface of our hNPs and delivery site specific therapeutics to diseases tissue.

OBJECTIVES/SPECIFIC AIMS: To identify new ethical challenges in human subject research related to the use of cloud-based platforms for data collection. METHODS/STUDY POPULATION: Ethical analysis. RESULTS/ANTICIPATED RESULTS: The OHRP regulations protecting the data, privacy and confidentiality of human subjects and the Terms of Service regulations governing data use by cloud-based platforms are vastly different. The gap between these 2 sets of laws and regulations leaves human subjects vulnerable to harm during the data collection process via clod-based tools. DISCUSSION/SIGNIFICANCE OF IMPACT: Recognizing the risks related to data gathering via cloud-based platforms, and educating researchers and research subjects about these risks and how to minimize them will strengthen the protections of participants and will enhance the informed consent process resulting in increased trust and greater willingness to participate in human subject research.

OBJECTIVES/SPECIFIC AIMS: Our objective is to study the role of soluble adenylyl cyclase in the melanocyte regulation of pigment in response to ultraviolet radiation. Melanocytes are specialized cells that produce melanin in organelles called melanosomes, and melanin determines the pigmentation of hair and skin. cAMP is a master regulator of pigmentation and transmembrane class of adenylyl cyclases are essential for expression of important enzymes involved in melanogenesis. However, pigmentation is also controlled by melanosomal pH, which regulates melanogenesis, tyrosinase activity, and melanosome maturation. The relationship between melanosomal pH and cAMP has been elusive. Soluble adenylyl cyclase is a noncanonical source of cAMP that is not responsive to G proteins but rather functions as a pH sensor. We recently demonstrated that loss of soluble adenylyl cyclase (sAC) activity leads to increased melanosomal pH as well as increased pigmentation in cells and hair. We expanded our research to investigate the role of sAC in the intrinsic response of melanocytes to ultraviolet radiation. METHODS/STUDY POPULATION: We utilized sACfl/fl (wild type) and sACKO mouse melanocytes and compared their change in pigmentation in response to ultraviolet radiation. Melanin was used as a measure of pigmentation. We irradiated these cells at differing doses of UVB (0, 1, 2, or 3 mJ/cm2) daily for 3 days. After UVB treatment, cells were observed and the surviving cell numbers were determined. Cells were then analyzed for melanin content using spectroscopy. RESULTS/ANTICIPATED RESULTS: We found that while both sACfl/fl and sACKO cells had increased melanin content in response to UVB, the melanin content of sACKO cells increased more compared with sACfl/fl cells (p=0.001 at daily dose of 3 mJ/cm2). In addition, sACKO cells required less UVB dose to induce a response. We also observed that sACKO cells show increased cell death compared with sACfl/fl cells. DISCUSSION/SIGNIFICANCE OF IMPACT: Although both sACfl/fl and sACKO cells can induce melanin production in response to UV, our results suggest that sACKO cells are more sensitive. We believe that this increased response in sACKO cells is due to increased melanosomal pH. In addition, sACKO cells show increased cell death, suggesting that sAC is important in the damage response secondary to UV exposure. UV plays a wide range of roles in skin biology such as contributing to cancer risk and pigmentation. Since pigmentation is essential for the protection of the skin from UV insult, further investigation of possible mechanisms in which sAC can influence pigmentation in response to UV is warranted.

OBJECTIVES/GOALS: The objective of this study is to define the molecular mechanisms that control survival of malignant stem cells in acute myeloid leukemia (AML). Leukemia stem cells (LSCs) are not effectively eradicated by standard treatment and lead to resistance and relapse, which contribute to poor survival rates. METHODS/STUDY POPULATION: The recently FDA approved venetoclax, a BCL2 inhibitor, with azacitidine, a hypomethylating agent leads to a 70% response rate in AML patients. Analysis of patients treated with this regimen showed direct targeting of LSCs. BCL2 has a non-canonical function in regulation of intracellular calcium. To determine how BCL2 mediated calcium signaling plays a role in LSC biology, we used LSCs isolated from venetoclax/azacitidine (ven/aza) sensitive and resistant patient samples to measure expression of calcium channels via RNA seq. BIO-ID, siRNA, flow cytometry, seahorse assays, calcium measurements and colony assays were used to determine the effects of calcium channel perturbation on LSC biology. RESULTS/ANTICIPATED RESULTS: BCL2 inhibition leads to decreased OXPHOS activity in primary AML specimens. BIO-ID studies revealed cation/metal ion transporters, ER membrane proteins and ER membrane organization as top enriched pathways interacting with BCL2. RNA-seq data showed increased expression of genes involved in calcium influx into the ER in ven/aza sensitive LSCs and increased expression of genes involved in calcium efflux from the ER in ven/aza resistant samples. Ven/Aza resistant LSCs have increased mitochondrial calcium content, consistent with their increased OXPHOS activity as calcium is required for OXPHOS. Perturbation of these channels leads to decreased OXPHOS activity and decreased viability in LSCs. DISCUSSION/SIGNIFICANCE OF IMPACT: We postulate that a deeper understanding of the mechanisms behind ven/aza targeting of LSCs will lead to the development of novel therapies for patients who do not respond to ven/aza. Our data show targeting intracellular calcium signaling could be a viable therapeutic strategy for AML patients.