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Published on: September 1, 2015
Inhibiting Focal Adhesion Kinase Ameliorates Cyst Development in Polycystin-1-Deficient Polycystic Kidney Disease in
Jinzhao He1,2, Shun Zhang1, Zhiwei Qiu1
1State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Peking University, Beijing, China.
Background:
Autosomal dominant polycystic kidney disease (ADPKD) is characterized by numerous cysts originating from renal tubules and is associated with significant tubular epithelial cell proliferation. Focal adhesion kinase (FAK) promotes tumor growth by regulating multiple proliferative pathways.
Methods:
We established the forskolin (FSK)-induced three-dimensional (3D) Madin-Darby Canine Kidney cystogenesis model and 8-bromoadenosine-3`,5`-cyclic monophosphate-stimulated cyst formation in ex vivo embryonic kidney culture. Cultured human renal cyst-lining cells (OX-161) and normal tubular epithelial cells were treated with FAK inhibitors or transfected with green fluorescent protein-tagged FAK mutant plasmids for proliferation study. Furthermore, we examined the role of FAK in two transgenic ADPKD animal models, the kidney-specific Pkd1 knockout and the collecting duct-specific Pkd1 knockout mouse models.
Results:
FAK activity was significantly elevated in OX-161 cells and in two ADPKD mouse models. Inhibiting FAK activity reduced cell proliferation in OX-161 cells and prevented cyst growth in ex vivo and 3D cyst models. In tissue-specific Pkd1 knockout mouse models, FAK inhibitors retarded cyst development and mitigated renal function decline. Mechanically, FSK stimulated FAK activation in tubular epithelial cells, which was blocked by a protein kinase A (PKA) inhibitor. Inhibition of FAK activation by inhibitors or transfected cells with mutant FAK constructs interrupted FSK-mediated Src activation and upregulation of ERK and mTOR pathways.
Conclusions:
Our study demonstrates the critical involvement of FAK in renal cyst development, suggests that FAK is a potential therapeutic target in treating patients with ADPKD, and highlights the role of FAK in cAMP-PKA-regulated proliferation.
Insights
Focal adhesion kinase (FAK) drives cyst growth in autosomal dominant polycystic kidney disease (ADPKD). Inhibiting FAK reduces proliferation and cyst development, suggesting FAK as a therapeutic target for ADPKD.
Area of Science:
- Nephrology
- Molecular Biology
- Cell Biology
Background:
- Autosomal dominant polycystic kidney disease (ADPKD) is characterized by renal tubule cysts and epithelial cell proliferation.
- Focal adhesion kinase (FAK) is implicated in promoting tumor growth via proliferative pathways.
Purpose of the Study:
- To investigate the role of FAK in ADPKD pathogenesis.
- To evaluate FAK inhibitors as a potential therapeutic strategy for ADPKD.
Main Methods:
- Established forskolin (FSK)-induced 3D cystogenesis and ex vivo embryonic kidney models.
- Utilized cultured human renal cyst-lining cells (OX-161) and ADPKD mouse models (Pkd1 knockout).
- Assessed FAK activity, cell proliferation, and cyst development following FAK inhibition or manipulation.
Main Results:
- FAK activity was elevated in ADPKD cells and models.
- FAK inhibition reduced cell proliferation and prevented cyst growth in vitro and in vivo.
- FAK inhibitors mitigated cyst development and improved renal function in ADPKD mouse models.
- FAK activation is downstream of cAMP-PKA signaling and upstream of Src, ERK, and mTOR pathways.
Conclusions:
- FAK plays a critical role in renal cyst development in ADPKD.
- Targeting FAK presents a potential therapeutic avenue for ADPKD treatment.
- FAK mediates cAMP-PKA-regulated proliferation in renal epithelial cells.
