Organoid Products & Services Core

PROJECT SUMMARY (CORE II - ORGANOID PRODUCTS & SERVICES) The goal of this PKD Core is to provide product and services enabling the use of human PKD organoids at the community-wide level by groups that would otherwise be unable to accomplish this for identification of therapeutic pathways and entities for treatment of PKD. Historically, there has been a lack of disease-specific phenotypic assays for PKD in vitro. This has limited the field's ability to discover therapies and predict responses. To address this gap, in 2015 we first described human PKD organoids that recapitulate pathophysiological features, including cystogenesis from kidney tubules. We have generated the world’s largest collection of iPS cells with relevant mutations in polycystins and cilia, alongside isogenic controls. Importantly, we have also demonstrated these cells to be capable of differentiation and expressing cystogenesis phenotypes in a disease-specific manner. We have further developed rigorous standard operating procedures for disease modeling, drug screening, and target validation. These efforts have established a working framework for understanding the PKD organoid phenotype, which other groups have utilized to reproduce the basic observations. While there is great interest in this technology, the system is costly, complex, and requires significant expertise to apply successfully, which makes it challenging to adopt. As a result, organoids are limited to a handful of laboratories. We will disrupt the paradigm by dramatically increasing the availability of iPS cells and samples thereof, and enabling outsourced screening services. In addition to the iPS cells themselves, we will also produce samples of processed organoids for target validation, and screening kits and services. We have developed a workflow wherein the first step is target validation to determine whether a therapeutic strategy or pathway is relevant to study in the PKD organoid system, followed by small-scale functional screening to determine whether an intervention in this pathway can produce a desired phenotypic rescue in organoids. In pilot screens, we have successfully applied this platform to gain novel insight into PKD pathophysiology, discover a therapeutic lead, and establish positive and negative controls. In addition to standard organoids, we will generate newly developed apical-in cystic organoids, with increased physiological relevance, as well as well-differentiated tubular epithelial cell lines with PKD mutations. We will utilize these tools to validate reagents generated by other Cores. We will pursue the following Specific Aims: (1) Generate well-characterized iPS cells and tubular epithelial cells that recapitulate phenotypes of ADPKD, ARPKD, and ciliopathies. (2) Efficiently validate mechanistic pathways and therapeutic interventions by enabling a community screening service. Target pathways and tools identified by other groups within the PKD RRC will be further validated and verified using our tools.