Biomechanical Forces and the Enteric Nervous System: Advancing the Development and Function of Tissue-Engineered Intestine - PROJECT SUMMARY/ABSTRACT Objectives: This application outlines a 5-year project to advance the field of intestinal tissue-engineering. Successful completion will enable this promising early stage investigator to conduct research to identify the processes required to generate functional tissue-engineered intestine as a cure for pediatric intestinal failure. Background: Intestinal failure is a survivable but morbid disease that afflicts tens of thousands of children. Contemporary management is inadequate and costly. Tissue-engineered intestine is a potential solution; however, complex intestinal functions have yet to be confirmed in existing human intestinal organoid (HIO) models. Thus, understanding the processes required to generate a functional intestine are a critical unmet need and a high NIH priority. The enteric nervous system (ENS) is the master regulator of bowel function. However, recent HIO models remain immature in ENS development, organization and function. Biomechanical forces are known to influence intestinal development; however, their role in ENS development and function in HIOs remains unknown. Based on published and preliminary data, our central hypothesis is that ENS development and function within HIOs depends on biomechanical force. Three aims will test this hypothesis: Research Design and Methods: HIO+ENS will be generated with and without endoluminal springs to apply mechanical strain. Aim 1 will define the role of biomechanical force on ENS spatial and cellular development in HIOs. Novel 3D imaging will evaluate ENS spatial organization and cellular diversity. Aim 2 will determine if biomechanical force enhances motility in HIOs. Ex vivo neurogenic contractility will be measured and IHC performed to assess the SIP syncytium, which regulates motility with the ENS. Aim 3 will ascertain if biomechanical force enhances ENS-dependent barrier function in HIOs. Transepithelial resistance, short circuit current, and permeability will be measured in an Ussing chamber. IHC will assess tight junctions. Aim 4 will employ snRNAseq to robustly characterize cell populations in HIOs (ENS, SIP syncytium, and tight junctions). This project is novel because no prior study has examined the role of biomechanical forces in ENS development and function in HIOs nor assessed ENS spatial development with novel 3D imaging methods. Career Development Plan & Goals: The investigator is a pediatric surgeon-scientist who is NIH-funded as a K08 awardee. She is now seeking her first R01 award as an Early Stage Investigator. Her career goals are to use this R01 to identify mechanistic pathways that can be leveraged to optimize tissue-engineered intestine generation for children with intestinal failure. Dr. Speer has long-standing collaborative and mentor relationships with Co-I’s Drs. Heuckeroth, Helmrath and Dunn, who will provide expertise and guidance on novel 3D imaging of the ENS, advanced HIO transplantation surgeries, and nitinol springs, respectively. Research Environment: The project will be carried out at UTHealth in the Texas Medical Center, the world’s largest medical center which seeks to nurture cross-institutional collaboration, creativity, and innovation.
