Ikeda Kisho, Kusaba Tetsuro, Tomita Aya, Watanabe-Uehara Noriko, Ida Tomoharu, Kitani Takashi, Yamashita Noriyuki, Uehara Masahiro, Matoba Satoaki, Yamada Tadaaki, Tamagaki Keiichi
Department of Nephrology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Department of Nephrology, Kyoto Prefectural University of Medicine, Kyoto, Japan,
Nephron. 2020;144(10):525-536. doi: 10.1159/000509419. Epub 2020 Aug 14.
The clinical features of autosomal dominant polycystic kidney disease (ADPKD) differ among patients even if they have the same gene mutation in PKD1 or PKD2. This suggests that there is diversity in the expression of other modifier genes or in the underlying molecular mechanisms of ADPKD, but these are not well understood.
We primarily cultured solute carrier family 12 member 3 (SLC12A3)-positive urine-derived distal tubular epithelial cells from 6 ADPKD patients and 4 healthy volunteers and established immortalized cell lines. The diversity in receptor tyrosine kinase (RTK) phosphorylation by phospho-RTK array in immortalized tubular epithelial cells was analyzed.
We noted diversity in the activation of several molecules, including Met, a receptor of hepatocyte growth factor (HGF). Administration of golvatinib, a selective Met inhibitor, or transfection of small interfering RNA for Met suppressed cell proliferation and downstream signaling only in the cell lines in which hyperphosphorylation of Met was observed. In three-dimensional culture of Madin-Darby canine kidney (MDCK) cells as a cyst formation model of ADPKD, HGF activated Met, resulting in an increased total cyst number and total cyst volume. Administration of golvatinib inhibited these phenotypes in MDCK cells.
Analysis of urine-derived tubular epithelial cells demonstrated diverse RTK phosphorylation in ADPKD, and Met phosphorylation was noted in some patients. Considering the difference in the effects of golvatinib on immortalized tubular epithelial cells among patients, this analysis may aid in selecting suitable drugs for individual ADPKD patients.
常染色体显性多囊肾病（ADPKD）患者的临床特征存在差异，即便他们在PKD1或PKD2基因上具有相同的基因突变。这表明在其他修饰基因的表达或ADPKD潜在分子机制中存在多样性，但目前对此了解尚少。
我们原代培养了6例ADPKD患者和4名健康志愿者的溶质载体家族12成员3（SLC12A3）阳性尿源远端肾小管上皮细胞，并建立了永生化细胞系。分析了永生化肾小管上皮细胞中磷酸化受体酪氨酸激酶（RTK）阵列对RTK磷酸化的多样性。
我们注意到包括肝细胞生长因子（HGF）受体Met在内的几种分子的激活存在多样性。给予选择性Met抑制剂戈伐替尼或转染针对Met的小干扰RNA仅在观察到Met过度磷酸化的细胞系中抑制细胞增殖和下游信号传导。在作为ADPKD囊肿形成模型的Madin-Darby犬肾（MDCK）细胞的三维培养中，HGF激活Met，导致囊肿总数和囊肿总体积增加。给予戈伐替尼可抑制MDCK细胞中的这些表型。
对尿源肾小管上皮细胞的分析表明ADPKD中存在多样的RTK磷酸化，且在部分患者中观察到Met磷酸化。考虑到戈伐替尼对不同患者永生化肾小管上皮细胞作用的差异，该分析可能有助于为个体ADPKD患者选择合适的药物。
