OBJECTIVES/GOALS: In Sep 2022, Johns Hopkins Research Coordinator Support Service launched Research Personnel Onboarding Program. The program on board in experienced individuals in 6-8 weeks, tailoring training plans to Investigator and study needs. It also offers Ongoing Support to enhance sustainability and adaptability. METHODS/STUDY POPULATION: * Assess Principal Investigator (PI)’s need Evaluate study’s need Understand trainee’s background Develop a personalized training plan (∼6-8 weeks) Weekly updates Ongoing mentorship Research staff spend 200+ training hours, depending on their need. Training encompasses various modalities: Interactive 1:1 onboarding sessions, Online, and Instructor-led trainings and sessions cover a wide range of topics, including: * Mandatory JHU/Institutional Review Board trainings * Good Clinical Practice * Regulatory submissions * Screening/Consenting * Monitoring/Auditing * Visit conduct * Clinical skills * Soft Skills Figure 1. Chart shows cumulative onboarding hours that focus on “How” to do tasks Figure 2. Chart shows cumulative training hours that focus on regulations, ethics and “Why” for tasks RESULTS/ANTICIPATED RESULTS: * The program contributed nearly 4000 hrs. of research staff training in the past 1 year * The program received 26 requests from investigators; 14 Completed the onboarding program, 1 Active, 5 Projected (Future start date), and 6 Cancelled (HR issues, lack of fund, or hired a trained staff) * 22 requests opted in the “Ongoing Support” * Ongoing Support, is averaged at 1 hr./month for the first 3-6 months. This indicates program success in empowering independent task performance * Developing REDCap request had significantly reduced meetings and paperwork * Web-Based Clockify invoicing has drastically reduced monthly manually invoicing processing time DISCUSSION/SIGNIFICANCE: * Grow the next generation of clinical research professionals * Centralize and standardize expert onboarding throughout the University * Improve outcomes, enhanced productivity, knowledge sharing, collaboration, and innovation * Decrease frustration and enhance satisfaction of trainees and departments

OBJECTIVES/SPECIFIC AIMS: This study attempts to evaluate the drinking patterns and traits of individuals who partake in high intensity drinking, defined as binge drinking at 2 or more times the minimum binge count (4 drinks for females, 5 drinks for males). METHODS/STUDY POPULATION: We analyzed data from non-treatment seeking volunteers enrolled in NIAAA screening protocols. The sample included 706 males and 474 females ranging in age from 18 to 91. Subjects were assigned to one of four groups (Non-Binge, Level 1, Level 2, Level 3) based on the highest binge session reported in their Timeline Followback questionnaire. The criteria for each group were different for males and females based on the current NIAAA definitions of binge drinking. The cutoffs for females were 0-3 drinks for Non-Binge, 4-7 drinks for Level 1, 8-11 drinks for Level 2, and 12+ drinks for Level 3. The male drink cutoffs were 0-4, 5-9, 10-14, and 15+ respectively. We looked at various drinking measures (Timeline Followback, Self-Reported Effects of Alcohol (SRE), Alcohol Use Disorders Identification Test (AUDIT)) and trait measures (UPPS-P Impulsivity Scale, Barratt’s Impulsiveness Scale, Buss Perry Aggression Questionnaire) to identify mean differences between groups. RESULTS/ANTICIPATED RESULTS: There were significant differences in drinking patterns between the groups for both males and females. Number of drinking days, average drinks per drinking day, and number of heavy drinking days all increased as binge level increased. There were also significant differences between groups in males for trait measures. Level 2 and Level 3 bingers scored significantly higher on impulsivity and aggression than the Level 1 and Non-Binge groups. Ongoing analyses are examining differences among binge groups on other measures including SRE and AUDIT. Future analyses will explore potential mechanisms underlying the relationships between trait measures and binge drinking using structural equation modeling. DISCUSSION/SIGNIFICANCE OF IMPACT: This study found significant differences between high-intensity drinkers, or “super bingers”, and lighter binge and non-binge drinkers. Super bingers showed an overall heavier drinking pattern across measures. The elevated aggression, impulsivity, and overall heavy drinking patterns of super bingers suggest a behavioral profile that makes this group in particular at higher risk for developing alcohol use disorder and related problems. These traits and behaviors may also help identify targets for treatment interventions for alcohol use disorder.

Minority populations are largely absent from clinical research trials. The neglect of these populations has become increasingly apparent, with escalating cancer burdens and chronic disease. The challenges to recruitment of minorities in the United States are multiple including trust or lack thereof. Keys to successful recruitment are responding to community issues, its history, beliefs, and its social and economic pressures. The strategy we have used in many low-income, sometimes remote, communities is to recruit staff from the same community and train them in the required basic research methods. They are the first line of communication. After our arrival in the Texas Rio Grande Valley in 2001, we applied these principles learned over years of global research, to studies of chronic diseases. Beginning in 2004, we recruited and trained a team of local women who enrolled in a cohort of over five thousand Mexican Americans from randomly selected households. This cohort is being followed, and the team has remained, acquiring not only advanced skills (ultrasound, FibroScan, retinal photos, measures of cognition, etc.) but capacity to derive key health information. Currently, we are participating in multiple funded studies, including an NIH clinical trial, liver disease, obesity, and diabetes using multiomics aimed at developing precision medicine approaches to chronic disease prevention and treatment.

OBJECTIVES/GOALS: The CTSI Research Infrastructure Network (RIN) expands CTSI’s reach into the regional health systems to provide our services to a broad and diverse translational science community. We create and support research collaborations that span multiple geographies and patient populations and serve as a bridge between the affiliate sites and CTSI programs. METHODS/STUDY POPULATION: We conducted needs assessments at each site (n=6) via in-depth, semi-structured interviews with key stakeholders. Informants (n=40) included investigators, study personnel, and research administrators. Investigators were selected across a variety of departments and career stages. Interview transcripts and notes were analyzed using matrix-based qualitative methods to identify both the common and unique research infrastructure needs of each site. Individualized support plans were shared with each site and a comprehensive summary report was presented to CTSI leadership. RIN met with UCSF’s Comprehensive Cancer Center, which conducts clinical trials at 2 sites, to coordinator our effort and services. When possible, RIN addressed service requests in real time that arose during interviews. RESULTS/ANTICIPATED RESULTS: We identified heterogeneous needs across multiple sites. However, among the community health systems with non-academic clinicians, there were common needs for research training, consultations in biostatistics/study design, and finding academic collaborators. The needs of sites with UCSF academic faculty differed from those of community sites and mainly included improved awareness and access to CTSI programs, ease of use of data extraction services, training programs, and assistance with regulatory approvals. Site needs are best addressed with individual plans created with CTSI Program leaders. A developing governance structure will include representation on a CTSI advisory committee and an annual conference to facilitate the sharing of best practices and foster collaboration across member sites. DISCUSSION/SIGNIFICANCE: Providing an individualized, site-specific approach to expanding CTSI services to regional health systems, will increase research collaborations across Northern California through building relationships, addressing unique infrastructure needs and sharing best practices throughout the network.

OBJECTIVES/GOALS: The Oregon Clinical and Translational Research Institute (OCTRI) Clinical Research Navigator program provides a single point of entry for clinical and translational research services, support, advice and guidance. We provide data to illustrate the Navigator model at OHSU and examine continued opportunities to optimize research resources. METHODS/STUDY POPULATION: Requests and activities performed by the OCTRI Navigator program, staffed by 3 FTE (2 Assistant Navigators and 1 Assistant Director) were analyzed. Navigator receives requests through multiple methods: a digital form (REDCap®), email, phone calls. Requests for services and support include focused need for a core or a broad request for multiple services for start-up: informatics, the clinical and translational research center, regulatory knowledge and support, recruitment, qualitative methods, community research, biostatistics or broad consultations. Requests are tracked in SPARCRequest. Navigator also supports wayfinding to institutional resources outside of the CTSA, matchmaking for sponsors seeking investigators, and serves as a connector and facilitator across programs. RESULTS/ANTICIPATED RESULTS: OCTRI Clinical Research Navigator triaged an average of 964 research requests for 613 projects with 388 unique investigators annually between 2018-2022. Navigator also fields more than 80 calls each year that are unrelated to CTSA projects. Project requests are examined to illustrate trends in projects requesting multiple services and display how Navigator simplifies project intake and connects researchers to resources they may have not recognized they needed. Project attributes including funding type and funding status are included in this review. DISCUSSION/SIGNIFICANCE: CTSA resources are essential to the infrastructure available to researchers. While absolute numbers of requests provide little insight into the impact each CTSA hub may have, the timing and clustering trends of projects with multiple program requests shows how a combination of technology and experienced staff can efficiently support researchers.

OBJECTIVES/GOALS: Having a strong leader in the form of an operations manager is crucial to lead and motivate teams. They're an additional set of problem-solving eyes to identify and address challenges. This poster demonstrates coordination techniques involved to work with teams of researchers, ensuring effective communication, deadlines met, and impact. METHODS/STUDY POPULATION: Stories, course corrections, publications, presentations and other metrics related to various projects and studies in the Lab will show such as the National Drug Early Warning System (NDEWS), Program to Alleviate National Disparities in Ethnic and Minority Immunizations in the Community (PANDEMIC, CDC), Polysubstance Use Study (PSU), and All of Us Consortium of CTSA/PACER Community Network. Additional data will be collected with project managers and students from each respective project to evaluate the effectiveness of an Operations Manager on the team across all studies in Our Lab. RESULTS/ANTICIPATED RESULTS: By sharing these results, showing the benefit to having an Operations Manager and the stories collected and shared for this poster, course corrections will set an example for future operations managers and their teams to continue to optimize efforts. Tis Could be a Special Interest Group (SIG)-related effort that would facilitate success to many labs. DISCUSSION/SIGNIFICANCE: Operations management focuses on effectively managing resources, ensuring that we meet deadlines, reduce delays, and maximize team productivity. By sharing our experiences, this can be a more standard practice among research labs to have smooth operations, and increase overall reputation of our team among those in the field.

OBJECTIVES/SPECIFIC AIMS: Glioblastoma (GBM) is a brain cancer with a devastatingly short overall survival of under two years. The poor prognosis of GBM is largely due to cell invasion and maintenance of cancer initiating cells that evade the brain’s innate and adaptive immune responses which enables escape from surgical resection and drives inevitable recurrence. While targeting the brain’s immune microenvironment has long been proposed as a strategy for treating GBM, translational progress has been slow, underscoring the need to investigate the brain’s immune microenvironment for therapeutic avenues. METHODS/STUDY POPULATION: Recent advancements in tunable synthetic immunomodulatory gene circuits targeting metastatic cancers has demonstrated the novel ability to use engineering principles to induce infiltrative cancer cells to express combinatorial immunomodulatory outputs that enable T-cell killing4. Our central hypothesis is: we will be able to significantly improve survival with a lasting immune-mediated control of GBM by using synthetic immunomodulatory gene circuits driving GBM cells to express a local combination of immunomodulatory proteins: human IL15, a surface T-cell engager, PD-L1-CD3 bispecific antibody, and the protein, LIGHT (TNFRSF14). Importantly, the co-expression of LIGHT and anti-PD-L1 therapies was recently shown to rescue PD-L1 checkpoint blockage in the preclinical models of brain tumors and significant enhance survival outcomes highlighting the benefits of novel combinations of immunomodulatory proteins for treatment of GBM. To identify genes whose expression is dramatically upregulated in GBM compared to normal human brain cells, a pooled of six thousand lentiviral oncogene promoters that drive expression of a red-fluorescent protein has been infected into three human GBM cell lines. RESULTS/ANTICIPATED RESULTS: We have successfully infected our GBM cells and are preparing samples for next generation DNA sequencing to determine highly active promoters in GBM that are not expressed in multiple normal brain cells types, astrocytes and neurons. These chosen promoters will then be used to drive an AND gate logic gene circuit immunotherapy outputs which is currently under development for both in-vitro and in-vivo experiments. DISCUSSION/SIGNIFICANCE OF IMPACT: We anticipate that local expression of multiple immune effectors proteins will significantly enhance tumor control and survival in both synergistic murine and human-murine xenograft pre-clinical models of GBM. Ultimately, our goal is to rapidly translate this technology advance into the clinical trial for adult GBM patients.

OBJECTIVES/SPECIFIC AIMS: Objective: Approximately 86 million people in the US have prediabetes, but only a fraction of them receive proven effective therapies to prevent diabetes. Further, the effectiveness of these therapies varies with individual risk of progression to diabetes. We estimated the value of targeting those individuals at highest diabetes risk for treatment, compared to treating all individuals meeting inclusion criteria for the Diabetes Prevention Program (DPP). METHODS/STUDY POPULATION: METHODS: Using a micro-simulation model, we estimated total lifetime costs and quality-adjusted life expectancy (QALE) for individuals receiving: (1) lifestyle intervention involving an intensive program focused on healthy diet and exercise, (2) metformin administration, or (3) no intervention. The model combines several components. First a Cox proportional hazards model predicted onset of diabetes from baseline characteristics for each pre-diabetic individual and yielded a probability distribution for each alternative. We derived this risk model from the Diabetes Prevention Program (DPP) clinical trial data and the follow-up study DPP-OS. The Michigan Diabetes Research Center Model for Diabetes then estimated costs and outcomes for individuals after diabetes diagnosis using standard of care diabetes treatment. Based on individual costs and QALE, we evaluated NMB of the two interventions at population and individual levels, stratified by risk quintiles for diabetes onset at 3 years. RESULTS/ANTICIPATED RESULTS: Results: Compared to usual care, lifestyle modification conferred positive benefits for all eligible individuals. Metformin’s NMB was negative for the lowest population risk quintile. By avoiding use among individuals who would not benefit, targeted administration of metformin conferred a benefit of $500-$800 per person, depending on duration of treatment effect. When treating only 20% of the population (e.g., due to capacity constraints), targeting conferred a NMB of $14,000-$18,000 per person for lifestyle modification and $16,000-$20,000 for metformin. DISCUSSION/SIGNIFICANCE OF IMPACT: Conclusions: Metformin confers value only among higher risk individuals, so targeting its use is worthwhile. While lifestyle modification confers value for all eligible individuals, prioritizing the intervention to high risk patients when capacity is constrained substantially increases societal benefits.

Understanding patient and caregiver experiences is a critical component of the conception, design, and implementation of clinical research studies. The “Database of Individual Patient Experiences” (DIPEx) is an innovative, evidence-based approach for eliciting rich information about health experiences. We conducted a formative evaluation with 14 pediatric oncology researchers to assess the value of using data from a DIPEx study on patient and caregiver experiences with childhood cancer to inform patient-centered research in pediatric oncology. Participants identified barriers to incorporating patient perspectives and experiences into their research and how the DIPEx approach could be leveraged to facilitate this practice.

The Accelerating COVID-19 Therapeutic Interventions and Vaccines (ACTIV) Cross-Trial Statistics Group gathered lessons learned from statisticians responsible for the design and analysis of the 11 ACTIV therapeutic master protocols to inform contemporary trial design as well as preparation for a future pandemic. The ACTIV master protocols were designed to rapidly assess what treatments might save lives, keep people out of the hospital, and help them feel better faster. Study teams initially worked without knowledge of the natural history of disease and thus without key information for design decisions. Moreover, the science of platform trial design was in its infancy. Here, we discuss the statistical design choices made and the adaptations forced by the changing pandemic context. Lessons around critical aspects of trial design are summarized, and recommendations are made for the organization of master protocols in the future.

OBJECTIVES/GOALS: A Mayo Clinic in Florida committee completes 100+ Scientific Reviews annually through manual e-mail and Excel tracking, placing a manual burden on reviewers and coordinators. REDCap, an electronic data capture system, was leveraged to reduce the administrative burden.#_ftn1 #_ftnref2 METHODS/STUDY POPULATION: Historically, emails were sent by a coordinator to physicians, requesting their initial review and following up with reminders. This process was tracked using Excel, presenting a need to make this process more efficient, so a workgroup was created. To ensure all perspectives were accounted for, the workgroup included the review coordinator, a physician reviewer, and study team members who submit development requests for studies. The process was mapped using the existing Excel spreadsheet, and email templates. Critical data elements were identified, ensuring the database would identify bottlenecks. Two REDCap instruments were then created: one to outline the coordinator workflow and a survey for physician reviewers to complete the scientific reviews. #_ftn2#_ftnref1 RESULTS/ANTICIPATED RESULTS: The workflow is live in REDCap and has effectively processed over 100 scientific reviews in <1 year. The system captures the review status and guides the coordinator through the workflow, capturing dates when tasks are completed. When review criteria are met, the database sends an email to the assigned reviewer. The email includes a link to a REDCap survey, containing all pertinent documents. The reviewer uploads their completed review form within the survey, if this is not completed within a given period, the database sends email reminders. Once the review is complete, a notification is sent to the review coordinator. The review workflow is accessible to the study team who requested scientific review, making them aware of their request status ad giving them access to the review the moment it is completed. DISCUSSION/SIGNIFICANCE: Leveraging REDCap has increased visibility, reduced overall manual processes, and simplified the reviewer burden by providing all the information needed in a single notification. REDCap is a cost-effective, impactful solution to simplifying administrative burden in managing committee reviews.

OBJECTIVES/SPECIFIC AIMS: Suggestibility, defined as the inclination to accept and internalize messages, has not been assessed much in relation to alcohol use. Prior research has shown that suggestibility to social cues and peer influence may play a role in driving alcohol consumption. Our previous work has shown associations between suggestibility and alcohol consumption in social drinkers. This study aims to examine how suggestibility and social susceptibility are related to ideas alcohol consumption and consequences across the spectrum of alcohol use and misuse. We hypothesize that those with higher suggestibility and social susceptibility reports will also have higher alcohol consumption and consequences, and that the impact of susceptibility is lower in dependent compared to non-dependent drinkers. METHODS/STUDY POPULATION: Study participants enrolled in the NIAAA screening and assessment protocol (N=157) completed questionnaires on suggestibility and alcohol consumption, The Multidimensional Iowa Suggestibility Scale (MISS) is a 95-question self-report assessment of suggestibility which draws from subcategories of consumer suggestibility, perceivability, physiological suggestibility, physiological reactivity, and peer conformity. Alcohol measures included 90-day Timeline Followback interviews and the Alcohol Use Disorder Identification Test (AUDIT). Participants also underwent the Structured Clinical Interviews for DSM-IV or DSM-5 disorders, and were stratified into two groups: alcohol dependent (N = 86) and non-dependent (N=71). Median split by age was additionally used to explore age’s relationship with suggestibility and alcohol with the under 36 (N = 45) and over 36 (N = 26) non-dependent groups. RESULTS/ANTICIPATED RESULTS: Initial analyses showed marked differences between the dependent and non-dependent groups in the relationship between the MISS total score and AUDIT total score. The non-dependent group showed significant positive correlations between MISS and AUDIT scores (r = 0.460, p < 0.001), while the dependent group did not show any correlation between MISS and AUDIT scores. Further examination of these relationships in the nondependent group revealed that MISS scores were also significantly positively correlated with AUDIT subscores of consumption, harm, and dependence. Age was found to have a significant negative correlation with MISS score (r = −0.354, p < 0.01). To better understand the role of age, the sample was split based on the median age (36 yrs), and analyzed separately. Results indicated robust relationships between MISS score and AUDIT (r = 0.457, p < 0.01) in the younger age group. In addition, the younger age group also showed significant relationships between MISS score and 90-day TLFB measures of total drinks, days drinking, and heavy drinking days. DISCUSSION/SIGNIFICANCE OF IMPACT: In non-dependent individuals, there was a significant positive relationship between suggestibility and alcohol measures, and these effects were amplified in younger individuals. No relationship was found between suggestibility and alcohol measures in the alcohol dependent individuals. This may be related to a greater impact of social and external contextual cues in younger social drinkers compared to dependent drinkers where tolerance and craving may have greater impact on alcohol consumption. These findings have important implications for traits that may put individuals at risk for developing harmful patterns of alcohol use and misuse. Future analyses will aim to examine biobehavioral underpinnings of the relationship between suggestibility and alcohol consumption.

OBJECTIVES/GOALS: Seeking ways to support teams in the preparation for and the implementation of the new National Institutes of Health (NIH) Policy for Data Management and Sharing (DMSP), the integrated Translational Health Research Institute of Virginia (iTHRIV) partnered with the UVA Health Sciences Library to develop training and resources for researchers. METHODS/STUDY POPULATION: Health sciences librarians and iTHRIV (an NIH-NCATS supported Clinical Translational Research Institute) convened a Working Group, inviting representatives from central and unit-specific research support offices (e.g. the Comprehensive Cancer Center), research compliance, regulatory affairs, sponsored programs, institutional review boards, libraries, and data science to review and discuss the DMSP requirements. After an initial orientation to the policy, the group reviewed existing public resources and solicited feedback about steps to best support UVA researchers in compliance. Leveraging the broad expertise of the group, the team provides guidance to researchers on writing the DMS plan and choosing a data repository, and provides tools and templates to support implementation of the policy. RESULTS/ANTICIPATED RESULTS: A library-created website provided policy guidance, including links to NIH-hosted information, resources created by other institutions, and new UVA-specific templates and suggested proposal language. Librarians led a webinar on the new policy and UVA resources which included a speaker from UVA regulatory affairs to describe the new DMSP requirements, and a tour of the new guide. The guide has been viewed over 5000 times to date and librarians have provided consultations and training to individuals and departments. Current plans include developing a user satisfaction survey, reviewing DMSP feedback from submitted proposals, and incorporating lessons learned into the website and future training. DISCUSSION/SIGNIFICANCE: The collaboration between iTHRIV and the Health Sciences Library to support the NIH Data Management and Sharing Policy was a successful partnership that provided leadership at the institutional level to communicate with and engage researchers and utilized the library’s web presence, expertise, and service model to provide direct support.

OBJECTIVES/GOALS: Identify causes for clinical research professional turnover Define data collection methods for exit interviews Provide institutions with resources to collect and analyze exit interviews Employ strategies to maximize the impact of exit interviews on retention METHODS/STUDY POPULATION: The Clinical Research Professional Taskforce (CRPT) exit interview Subgroup has met monthly since January 2023. Action items were agreed to and minutes were kept and reviewed at subsequent virtual working meetings. All members were given opportunity to speak and contribute. After a landscape analysis, conducted via survey, five institutions agreed to provide examples of their exit interview questions. Members spoke at length about goals, methods, collection techniques, institutional involvement, lessons learned and practical applications that could become best practices. RESULTS/ANTICIPATED RESULTS: The Subgroup aggregated all questions into categories and developed sample questions incorporating all data without using any word for word. In order to allow for quantitative assessment and standardized reporting the Subgroup formulated questions to be responded to utilizing a Likert scale with free text fields for select questions where further information is needed. The Subgroup developed best practices describing decision-making metrics, understanding reasons for turnover and reporting data back to leadership. Practical aspects such as method and time of survey collection, anonymity, and training staff are also included. DISCUSSION/SIGNIFICANCE: We are hopeful that sample questions and best practices will be helpful and widely utilized. Understanding the causes and impacts of CRP turnover are critical to meeting the current needs of clinical research. Further work is being done to calculate the cost of turnover to make the business case.

OBJECTIVES/SPECIFIC AIMS: The goal of this project is to study fetal pulmonary vasculature in a CDH animal model, to understand how FETO affects developing vasculature, and to develop a modifiable fetal tracheal occlusive therapeutic device that avoids previously seen sequelae of FETO, like alveolar distension, decreased surfactant production, and decreased Type II Pneumocytes. The primary outcome is lung volume/kilogram. The secondary outcomes are contrast-enhanced ultrasound perfusion metrics (Time to Peak, Mean Transit Time, Wash-in Rate, Wash-in Perfusion Index), pulmonary vascular density, Lung Injury Histology Scores, and Lung Compliance upon ventilation. METHODS/STUDY POPULATION: Congenital diaphragmatic hernias will be modeled by surgical hernia creation via maternal laparotomy and hysterotomy at gestational age 72 - 74 days. The ewe will undergo a second laparotomy at 105 - 115 days gestational age. After a second hysterotomy is made, the fetus will be removed from the amniotic sac, though placental circulation will be maintained (EXIT Procedure). The animal is cannulated via the umbilical vein and arteries onto the pumpless ECMO circuit. The balloon and pressure sensor complex is placed into the trachea via direct laryngoscopy, and the fetus aseptically sealed into the Biobag. The wires of the tracheal occlusive device (balloon catheter and pressure sensor) will egress via the port of the Biobag. The fetus remains in the Biobag for fourteen days, with the tracheal occlusive device in place for ten days, followed by a four day recovery period. Daily contrast-enhanced ultrasounds and pulmonary artery dopplers are performed. Upon study completion, the fetus is intubated and placed on a conventional ventilator. A full necropsy is then performed, with perfusion fixation of the lungs via the pulmonary artery. RESULTS/ANTICIPATED RESULTS: Hypothesis 1: Modifiable Tracheal Occlusion will have statistically different effects on developing lung parenchyma, surfactant production, and abundance of Type II Pneumocytes Hypothesis 2: Modifiable Tracheal Occlusion will have lower levels of pulmonary hypertension than negative control animals, as measured by contrast-enhanced ultrasound (pulmonary artery velocity and washout time). DISCUSSION/SIGNIFICANCE OF IMPACT: This project will provide insight into the development of pulmonary hypertension in the CDH fetus. It will provide insight into the physiology of FETO, a novel therapy for congenital diaphragmatic hernias, and will demonstrate the utility of the EXTEND System for fetal treatments that are not possible in the maternal uterus.