Rendu John, Montjean Rodrick, Coutton Charles, Suri Mohnish, Chicanne Gaetan, Petiot Anne, Brocard Julie, Grunwald Didier, Pietri Rouxel France, Payrastre Bernard, Lunardi Joel, Dorseuil Olivier, Marty Isabelle, Fauré Julien
Cellular Myology and Pathology, INSERM, U1216, Grenoble, France.
Grenoble Institut of Neurosciences, Université Grenoble Alpes, France.
Hum Mutat. 2017 Feb;38(2):152-159. doi: 10.1002/humu.23139. Epub 2016 Nov 21.
Dent-2 disease and Lowe syndrome are two pathologies caused by mutations in inositol polyphosphate 5-phosphatase OCRL gene. Both conditions share proximal tubulopathy evolving to chronic kidney failure. Lowe syndrome is in addition defined by a bilateral congenital cataract, intellectual disability, and hypotonia. The pathology evolves in two decades to a severe condition with renal complications and a fatal issue. We describe here a proof of principle for a targeted gene therapy on a mutation of the OCRL gene that is associated with Lowe syndrome. The affected patient bears a deep intronic mutation inducing a pseudo-exon inclusion in the mRNA, leading to a OCRL-1 protein loss. An exon-skipping strategy was designed to correct the effect of the mutation in cultured cells. We show that a recombinant U7-modified small RNA efficiently triggered the restoration of normal OCRL expression at mRNA and protein levels in patient's fibroblasts. Moreover, the PI(4,5)P2 accumulation and cellular alterations that are hallmark of OCRL-1 dysfunction were also rescued. Altogether, we provide evidence that the restoration of OCRL-1 protein, even at a reduced level, through RNA-based therapy represents a potential therapeutic approach for patients with OCRL splice mutations.
丹特2型疾病和洛氏综合征是由肌醇多磷酸5-磷酸酶OCRL基因突变引起的两种病症。这两种病症都有近端肾小管病变并发展为慢性肾衰竭。此外,洛氏综合征的特征还包括双侧先天性白内障、智力残疾和肌张力减退。该病症在二十年内会发展为伴有肾脏并发症的严重疾病并导致致命后果。我们在此描述了针对与洛氏综合征相关的OCRL基因突变进行靶向基因治疗的原理证明。受影响的患者存在一个深度内含子突变,该突变导致mRNA中包含一个假外显子,从而导致OCRL-1蛋白缺失。设计了一种外显子跳跃策略来纠正培养细胞中该突变的影响。我们表明,重组U7修饰的小RNA能有效促使患者成纤维细胞中mRNA和蛋白水平的正常OCRL表达得以恢复。此外,OCRL-1功能障碍的标志性特征——PI(4,5)P2积累和细胞改变也得到了挽救。总之,我们提供的证据表明,通过基于RNA的疗法恢复OCRL-1蛋白,即使水平降低,也代表了一种针对OCRL剪接突变患者的潜在治疗方法。