Inhibiting Focal Adhesion Kinase Ameliorates Cyst Development in Polycystin-1-Deficient Polycystic Kidney Disease in Animal Model
Inhibiting Focal Adhesion Kinase Ameliorates Cyst Development in Polycystin-1-Deficient Polycystic Kidney Disease in Animal Model
复制标题
抑制粘着斑激酶可改善多囊蛋白-1缺陷型多囊肾病动物模型中的囊肿发育
DOI：
10.1681/asn.2020111560
复制
发表时间：
2021-09-01
影响因子：
13.6
通讯作者：
Zhou, Hong
中科院分区：
文献类型：
--
作者：
He, Jinzhao;Zhang, Shun;Zhou, Hong
黏着斑激酶（FAK）已被广泛研究，并显示出对肿瘤进展具有至关重要的作用。虽然常染色体显性遗传性多囊肾病（ADPKD）与肿瘤有许多共同点，但FAK是否参与囊肿的发展尚不确定。我们的研究表明，使用小分子FAK抑制剂VS-4718抑制FAK活性可改善ADPKD进展并改善肾功能。抑制FAK及其下游Src信号传导可抑制ADPKD动物中cAMP介导的囊性上皮细胞异常增殖。这项研究强调了小分子FAK抑制剂在治疗ADPKD患者中的治疗潜力。背景常染色体显性遗传性多囊肾病（ADPKD）的特征是大量起源于肾小管的囊肿，并伴有显著的肾小管上皮细胞增殖。粘着斑激酶（FAK）通过调节多种增殖途径促进肿瘤生长。方法建立毛喉素（Forskolin，FSK）诱导的Madin-Darby犬肾三维（3D）囊肿形成模型和8-溴腺苷-3 '，5'-环磷酸（8-bromoadenosine-3 '，5'-cyclicmonophosphate，8-bromoadenosine-3 '，5'-cyclicmonophosphate，8-bromoadenosine-3 '，5'-cyclicmonophosphate）诱导的离体胚肾囊肿形成模型。用FAK抑制剂处理培养的人肾囊肿衬里细胞（OX-161）和正常肾小管上皮细胞，或用绿色荧光蛋白标记的FAK突变体质粒转染用于增殖研究。此外，我们研究了FAK在两种转基因ADPKD动物模型中的作用，即肾特异性Pkd 1敲除和集合管特异性Pkd 1敲除小鼠模型。结果在OX-161细胞和两种ADPKD小鼠模型中FAK活性均显著升高。抑制FAK活性减少OX-161细胞中的细胞增殖，并防止离体和3D囊肿模型中的囊肿生长。在组织特异性Pkd 1基因敲除小鼠模型中，FAK抑制剂延缓了囊肿的发展并减轻了肾功能下降。在机械上，FSK刺激肾小管上皮细胞中的FAK活化，其被蛋白激酶A（PKA）抑制剂阻断。通过抑制剂或用突变FAK构建体转染的细胞抑制FAK活化，中断了FSK介导的Src活化和ERK和mTOR途径的上调。结论FAK在肾囊肿的发生发展中起重要作用，FAK是ADPKD治疗的潜在靶点，FAK在cAMP PKA调控的细胞增殖中起重要作用。
Visual Abstract Significance Statement Focal adhesion kinase (FAK) has been widely studied and shown to have a crucial effect on tumor progression. Although autosomal dominant polycystic kidney disease (ADPKD) shares numerous commonalities with tumors, it is undetermined whether FAK is involved in cyst development. Our study demonstrates that inhibiting FAK activity using small molecule FAK inhibitor VS-4718 ameliorates ADPKD progression and improves renal function. Inhibiting FAK and its downstream Src signaling suppresses cAMP-mediated aberrant proliferation of cystic epithelial cells in ADPKD animals. This study highlights the therapeutic potential of small molecule FAK inhibitors in treating patients with ADPKD. 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.