EXOCYST INACTIVATION IN UROTHELIAL CELLS LEADS TO FN14 UPREGULATION AND CELL DEATH IN A MOUSE MODEL OF PRENATAL URETER OBSTRUCTIONS

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dc.contributor.advisorFogelgren, Ben
dc.contributor.authorVilliger, Ross
dc.contributor.departmentCell and Molecular Biology
dc.date.accessioned2022-07-05T19:57:43Z
dc.date.issued2022
dc.description.degreePh.D.
dc.embargo.liftdate2022-12-25
dc.identifier.urihttps://hdl.handle.net/10125/102128
dc.subjectCellular biology
dc.subjectDevelopmental biology
dc.subjectMolecular biology
dc.subjectexocyst
dc.subjectFn14
dc.subjectureter
dc.subjecturothelium
dc.titleEXOCYST INACTIVATION IN UROTHELIAL CELLS LEADS TO FN14 UPREGULATION AND CELL DEATH IN A MOUSE MODEL OF PRENATAL URETER OBSTRUCTIONS
dc.typeThesis
dcterms.abstractCongenital obstructive nephropathy (CON) is the leading cause of chronic kidney disease(CKD) and end-stage renal disease in children. This disorder is usually detected as hydronephrosis by prenatal ultrasounds. The most common cause of CON is obstruction at the ureteropelvic junction (UPJ), where the renal pelvis transitions to the upper ureter. EXOC5 is the central stabilizing subunit of the exocyst, an octameric protein complex that regulates exocytosis of secretory vesicles to the plasma membrane. The Fogelgren laboratory previously generated a novel EXOC5-conditional knockout (EXOC5-CKO) mouse using the Cre/loxP technology to inactivate EXOC5 in ureteric bud-derived epithelial cells. The EXOC5-CKO mice had severe bilateral UPJ obstructions, hydronephrosis, and succumbed to neonatal death. In these mice, UPJ obstruction was preceded by failure of urothelial stratification and cell death at E17.5, followed by a fibroproliferative response by the underlying mesenchyme. By transcriptomic microarray, we identified the Fn14 gene as being robustly upregulated in the EXOC5-CKO embryonic ureter at E16.5. Fn14 is a transmembrane tumor necrosis factor (TNF) receptor superfamily member with a single known ligand, TWEAK, which can activate different pathways depending on cell context, including programmed cell death. We demonstrated that ureter obstructions can be rescued in EXOC5-CKO mice by administering a pan-caspase inhibitor zVAD-fmk at E16.5. We also found inhibited urothelial stratification alone did not drive cell death during ureter development. Instead, exocyst-perturbed urothelial cells in vivo and in vitro have disrupted autophagy which activated NF-κB signaling and cell death. To study the role of Fn14 in renal and urological diseases, we generated a global Fn14-knockout (KO) mouse using CRISPR-Cas9 gene editing. Fn14-KO mice had ameliorated renal pathology in a cisplatininduced CKD model. Our findings support the hypothesis that urothelial cell death is a critical event leading to UPJO pathogenesis and that Fn14 signaling contributes to cell death in renal and urological pathologies.
dcterms.extent135 pages
dcterms.languageen
dcterms.publisherUniversity of Hawai'i at Manoa
dcterms.rightsAll UHM dissertations and theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
dcterms.typeText
local.identifier.alturihttp://dissertations.umi.com/hawii:11377

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Ph.D. - Cell and Molecular Biology