Zhu Ping, Sieben Cynthia J, Xu Xiaolei, Harris Peter C, Lin Xueying
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA.
Division of Cardiovascular Diseases, Mayo Clinic, Rochester, MN, USA.
Hum Mol Genet. 2017 Jan 1;26(1):158-172. doi: 10.1093/hmg/ddw376.
Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in either PKD1 or PKD2. It is one of the most common heritable human diseases with eventual development of renal failure; however, effective treatment is lacking. While inhibition of mechanistic target of rapamycin (mTOR) effectively slows cyst expansions in animal models, results from clinical studies are controversial, prompting further mechanistic studies of mTOR-based therapy. Here, we aim to establish autophagy, a downstream pathway of mTOR, as a new therapeutic target for PKD. We generated zebrafish mutants for pkd1 and noted cystic kidney and mTOR activation in pkd1a mutants, suggesting a conserved ADPKD model. Further assessment of the mutants revealed impaired autophagic flux, which was conserved in kidney epithelial cells derived from both Pkd1-null mice and ADPKD patients. We found that inhibition of autophagy by knocking down the core autophagy protein Atg5 promotes cystogenesis, while activation of autophagy using a specific inducer Beclin-1 peptide ameliorates cysts in the pkd1a model. Treatment with compound autophagy activators, including mTOR-dependent rapamycin as well as mTOR-independent carbamazepine and minoxidil, markedly attenuated cyst formation and restored kidney function. Finally, we showed that combination treatment with low doses of rapamycin and carbamazepine was able to attenuate cyst formation as effectively as a single treatment with a high dose of rapamycin alone. In summary, our results suggested a modifying effect of autophagy on ADPKD, established autophagy activation as a novel therapy for ADPKD, and presented zebrafish as an efficient vertebrate model for developing PKD therapeutic strategies.
常染色体显性多囊肾病(ADPKD)由PKD1或PKD2突变引起。它是最常见的人类遗传性疾病之一,最终会发展为肾衰竭;然而,目前缺乏有效的治疗方法。虽然在动物模型中抑制雷帕霉素的作用靶点(mTOR)能有效减缓囊肿扩张,但临床研究结果存在争议,这促使人们对基于mTOR的治疗方法进行进一步的机制研究。在此,我们旨在将自噬(mTOR的下游途径)确立为ADPKD的新治疗靶点。我们构建了pkd1的斑马鱼突变体,并在pkd1a突变体中发现了肾囊肿和mTOR激活,这表明存在一个保守的ADPKD模型。对这些突变体的进一步评估显示自噬通量受损,这在源自Pkd1基因敲除小鼠和ADPKD患者的肾上皮细胞中是保守的。我们发现,通过敲低核心自噬蛋白Atg5来抑制自噬会促进囊肿形成,而使用特异性诱导剂Beclin-1肽激活自噬则可改善pkd1a模型中的囊肿。用包括mTOR依赖性雷帕霉素以及mTOR非依赖性卡马西平和米诺地尔在内的复合自噬激活剂进行治疗,可显著减轻囊肿形成并恢复肾功能。最后,我们表明低剂量雷帕霉素和卡马西平联合治疗能够像单独使用高剂量雷帕霉素单一治疗一样有效地减轻囊肿形成。总之,我们的结果表明自噬对ADPKD具有调节作用,确立了自噬激活作为ADPKD的一种新治疗方法,并将斑马鱼作为开发PKD治疗策略的高效脊椎动物模型。