Linn Aung Khine, Maneepitasut Warun, Tubsuwan Alisa, Kitiyanant Narisorn, Phakdeekitcharoen Bunyong, Borwornpinyo Suparerk, Hongeng Suradej, Phanthong Phetcharat
Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
Molecular Medicine Program, Multidisciplinary Unit, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
J Pers Med. 2022 May 9;12(5):766. doi: 10.3390/jpm12050766.
Autosomal dominant polycystic kidney disease (ADPKD) is one of the most prevalent genetic diseases affecting the kidneys. A genetically specific mutation model is required to comprehend its pathophysiology and to develop a drug treatment. In this study, we successfully developed human induced pluripotent stem cells (hiPSCs) named MUi027-A from skin fibroblasts of a patient diagnosed with ADPKD and carrying the PKD1 frameshift mutation (c.7946_7947delCT). MUi027-A cells showed the same genetic fingerprints as the parental cells, including the presence of the PKD1 mutation. MUi027-A hiPSCs displayed embryonic stem cell-like characteristics with the capability of differentiating into the three germ layers. Upon directed differentiation, MUi027-A hiPSCs could be differentiated into tubular organoids with the expression of renal cell markers. Furthermore, we compared the efficiency of cyst formation in two human iPSC lines with different PKD1 mutations. When cyst formation was induced by either forskolin or blebbistatin, MUi027-A hiPSC-derived kidney organoids displayed higher frequencies of cyst formation when compared to organoids generated from an iPSC cell line with non-truncating PKD1 mutation genotype (c.5878C > T), suggesting the presence of physiological differences in the mechanism of cyst formation between different PKD1 mutants. Overall, we generated and characterized a novel human iPSC line with a specific PKD mutation and demonstrated its potential as a disease model to study the pathophysiology of genetic determinants in the development of ADPKD disease.
常染色体显性多囊肾病（ADPKD）是影响肾脏的最常见的遗传疾病之一。需要一个基因特异性突变模型来理解其病理生理学并开发药物治疗方法。在本研究中，我们成功地从一名被诊断患有ADPKD并携带PKD1移码突变（c.7946_7947delCT）的患者的皮肤成纤维细胞中培养出名为MUi027 - A的人诱导多能干细胞（hiPSC）。MUi027 - A细胞显示出与亲代细胞相同的基因指纹，包括PKD1突变的存在。MUi027 - A hiPSC表现出胚胎干细胞样特征，具有分化为三个胚层的能力。在定向分化后，MUi027 - A hiPSC可以分化为表达肾细胞标志物的管状类器官。此外，我们比较了两种具有不同PKD1突变的人iPSC系中囊肿形成的效率。当用福斯高林或blebbistatin诱导囊肿形成时，与由具有非截断PKD1突变基因型（c.5878C > T）的iPSC细胞系产生的类器官相比，MUi027 - A hiPSC来源的肾类器官显示出更高的囊肿形成频率，这表明不同PKD1突变体之间囊肿形成机制存在生理差异。总体而言，我们生成并鉴定了一种具有特定PKD突变的新型人iPSC系，并证明了其作为疾病模型研究ADPKD疾病发生发展中遗传决定因素病理生理学的潜力。
