PDF(Pubmed)
Abstract：
In polycystic kidney disease (PKD), microscopic tubules expand into macroscopic cysts. Among the world\'s most common genetic disorders, PKD is inherited via heterozygous loss-of-function mutations but is theorized to require additional loss of function. To test this, we establish human pluripotent stem cells in allelic series representing four common nonsense mutations, using CRISPR base editing. When differentiated into kidney organoids, homozygous mutants spontaneously form cysts, whereas heterozygous mutants (original or base corrected) express no phenotype. Using these, we identify eukaryotic ribosomal selective glycosides (ERSGs) as PKD therapeutics enabling ribosomal readthrough of these same nonsense mutations. Two different ERSGs not only prevent cyst initiation but also limit growth of pre-formed cysts by partially restoring polycystin expression. Furthermore, glycosides accumulate in cyst epithelia in organoids and mice. Our findings define the human polycystin threshold as a surmountable drug target for pharmacological or gene therapy interventions, with relevance for understanding disease mechanisms and future clinical trials.
摘要：
在多囊肾病（PKD）中，微观小管扩张成宏观囊肿。在世界上最常见的遗传疾病中，PKD是通过杂合功能丧失突变遗传的，但理论上需要额外的功能丧失。为了测试这个，我们在等位基因系列中建立了人类多能干细胞，代表四种常见的无义突变，使用CRISPR基础编辑。当分化为肾脏类器官时，纯合突变体自发形成囊肿，而杂合突变体（原始或碱基校正）不表达表型。使用这些，我们鉴定了真核核糖体选择性糖苷(ERSGs)作为PKD治疗药物,能够实现这些相同无义突变的核糖体通读.两种不同的ERSG不仅可以预防囊肿的发生，而且还可以通过部分恢复多囊素的表达来限制预先形成的囊肿的生长。此外，糖苷在类器官和小鼠的囊肿上皮中积累。我们的发现将人类多囊素阈值定义为药物或基因治疗干预的可突破的药物靶标。与了解疾病机制和未来的临床试验相关。
