Renal and hepatic cysts along with fibrosis (Hepatorenal Fibrocystic Disease, HRFD) are phenotypes observed in a large number of human syndromes (PKD, MKS, NPHP, JBTS, and BBS), the most prevalent of which is PKD. Unfortunately, current treatments for HRFDs are largely palliative and new more efficacious strategies are needed to retard or reverse cyst progression. Animal models that develop HRFDs have been instrumental in advancing our understanding of the molecular and cellular basis of these disorders and also serve as preclinical platforms to test candidate therapeutic and means of intervention. One key theme derived from these models is that most mutations causing HRFDs occur in genes that encode cilia proteins or proteins that are required for cilia assembly or function. While animal models of HRFDs are essential tools for advancing the field, there are many hurdles that prevent their full utilization in many labs. This includes the expertise, cost, and effort required to set up and generate the models and their ready availability. It can often take a year to establish new models at an investigator's home institution, which is prohibitive for rapid testing of novel hypotheses. To help reduce these hurdles and to increase the pace of research into HRFDs, the UAB Engineered Models Core (Core B) will generate mutations in HRFD/cilia related genes in cell lines, C. elegans, zebrafish, and mice using CRISPR based strategies and through ES cells provided by the mouse targeted knockout consortia. The Core will also generate in vivo mouse and zebrafish models expressing human disease alleles. This will allow the assessment of the pathogenic consequence of missense mutations in cyst progression. To facilitate intravital analyses of pathways associated with cystic disease, Core B will generate several biosensor lines to evaluate changes in cytosolic and ciliary Ca2+ and cAMP levels as well as a CiliaRFP mouse to visualize and analyze cilia in live tissues in situ. It can often take up to a year to establish new models at an investigator's home institution which prevents the rapid testing of novel hypotheses. PKD, MKS, NPHP, JBTS, and BBS models made available through Core B's efforts will be essential resources for analyzing cystogenic pathways and in preclinical trails that will conducted by the user base through the Center's Cell Physiology Core (Core C) and the Therapeutic Screening and Development Core (Core D). Finally, Core B will benefit Center members by providing biological reagents (tissue samples, blood, urine, DNA, RNA, protein) derived from our HRFD mutant models. These resources will facilitate pilot studies to test innovative hypotheses, preliminary studies needed for grant submissions, and conformational studies of in vitro findings without requiring the line be established at an investigators home institution. Overall, these genetic tools and biological resources that will be made available through Core B will provide a comprehensive and integrated set of resources examining the role of cilia and HRFD genes in cyst pathogenesis and for analyzing new treatment strategies.
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