Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas.
Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, Kansas.
Am J Physiol Renal Physiol. 2022 Oct 1;323(4):F492-F506. doi: 10.1152/ajprenal.00095.2022. Epub 2022 Aug 18.
Autosomal dominant polycystic kidney disease (ADPKD) is a debilitating renal neoplastic disorder with limited treatment options. It is characterized by the formation of large fluid-filled cysts that develop from kidney tubules through abnormal cell proliferation and cyst-filling fluid secretion driven by cAMP-dependent Cl secretion. We tested the effectiveness of the indazole carboxylic acid H2-gamendazole (H2-GMZ), a derivative of lonidamine, to inhibit these processes using in vitro and in vivo models of ADPKD. H2-GMZ was effective in rapidly blocking forskolin-induced, Cl-mediated short-circuit currents in human ADPKD cells, and it significantly inhibited both cAMP- and epidermal growth factor-induced proliferation of ADPKD cells. Western blot analysis of H2-GMZ-treated ADPKD cells showed decreased phosphorylated ERK and decreased hyperphosphorylated retinoblastoma levels. H2-GMZ treatment also decreased ErbB2, Akt, and cyclin-dependent kinase 4, consistent with inhibition of heat shock protein 90, and it decreased levels of the cystic fibrosis transmembrane conductance regulator Cl channel protein. H2-GMZ-treated ADPKD cultures contained a higher proportion of smaller cells with fewer and smaller lamellipodia and decreased cytoplasmic actin staining, and they were unable to accomplish wound closure even at low H2-GMZ concentrations, consistent with an alteration in the actin cytoskeleton and decreased cell motility. Experiments using mouse metanephric organ cultures showed that H2-GMZ inhibited cAMP-stimulated cyst growth and enlargement. In vivo, H2-GMZ was effective in slowing postnatal cyst formation and kidney enlargement in the mouse model. Thus, H2-GMZ treatment decreases Cl secretion, cell proliferation, cell motility, and cyst growth. These properties, along with its reported low toxicity, suggest that H2-GMZ might be an attractive candidate for treatment of ADPKD. Autosomal dominant polycystic kidney disease (ADPKD) is a renal neoplastic disorder characterized by the formation of large fluid-filled cysts that develop from kidney tubules through abnormal cell proliferation and cyst-filling fluid secretion driven by cAMP-dependent Cl secretion. This study shows that the lonidamine derivative H2-GMZ inhibits Cl secretion, cell proliferation, and cyst growth, suggesting that it might have therapeutic value for the treatment of ADPKD.
常染色体显性遗传多囊肾病（ADPKD）是一种进行性肾脏肿瘤性疾病，治疗选择有限。它的特征是通过异常的细胞增殖和 cAMP 依赖性 Cl 分泌驱动的囊泡填充液分泌，从肾小管形成大的充满液体的囊肿。我们使用 ADPKD 的体外和体内模型来测试吲唑羧酸 H2-加美扎嗪（H2-GMZ），一种 lonidamine 的衍生物，抑制这些过程的有效性。H2-GMZ 可有效快速阻断福司可林诱导的人 ADPKD 细胞中的 Cl 介导的短路电流，并且显著抑制 ADPKD 细胞的 cAMP 和表皮生长因子诱导的增殖。用 H2-GMZ 处理的 ADPKD 细胞的 Western blot 分析显示磷酸化 ERK 和过度磷酸化视网膜母细胞瘤水平降低。H2-GMZ 处理还降低了 ErbB2、Akt 和细胞周期蛋白依赖性激酶 4，与热休克蛋白 90 的抑制一致，并且降低了囊性纤维化跨膜电导调节剂 Cl 通道蛋白的水平。用 H2-GMZ 处理的 ADPKD 培养物含有更高比例的较小细胞，这些细胞具有更少和更小的片状伪足，细胞质肌动蛋白染色减少，即使在低浓度的 H2-GMZ 下，它们也无法完成伤口闭合，这与肌动蛋白细胞骨架的改变和细胞迁移性降低一致。使用小鼠后肾器官培养物的实验表明，H2-GMZ 抑制 cAMP 刺激的囊肿生长和增大。在体内，H2-GMZ 可有效减缓小鼠模型中出生后的囊肿形成和肾脏增大。因此，H2-GMZ 治疗可减少 Cl 分泌、细胞增殖、细胞迁移和囊肿生长。这些特性以及其报道的低毒性表明，H2-GMZ 可能是治疗 ADPKD 的有吸引力的候选药物。常染色体显性遗传多囊肾病（ADPKD）是一种肾脏肿瘤性疾病，其特征是形成大的充满液体的囊肿，这些囊肿从肾小管通过异常的细胞增殖和 cAMP 依赖性 Cl 分泌驱动的囊泡填充液分泌而发展。本研究表明，lonidamine 衍生物 H2-GMZ 抑制 Cl 分泌、细胞增殖和囊肿生长，表明它可能对治疗 ADPKD 具有治疗价值。
