Université Pierre et Marie Curie-Paris 6, IFR14, Paris F-75013, France.
Hum Mol Genet. 2010 Dec 15;19(24):4820-36. doi: 10.1093/hmg/ddq413. Epub 2010 Sep 21.
Autosomal dominant centronuclear myopathy (AD-CNM) is due to mutations in the gene encoding dynamin 2 (DNM2) involved in endocytosis and intracellular membrane trafficking. To understand the pathomechanisms resulting from a DNM2 mutation, we generated a knock-in mouse model expressing the most frequent AD-CNM mutation (KI-Dnm2(R465W)). Heterozygous (HTZ) mice developed a myopathy showing a specific spatial and temporal muscle involvement. In the primarily and prominently affected tibialis anterior muscle, impairment of the contractile properties was evidenced at weaning and was progressively associated with atrophy and histopathological abnormalities mainly affecting mitochondria and reticular network. Expression of genes involved in ubiquitin-proteosome and autophagy pathways was up-regulated during DNM2-induced atrophy. In isolated muscle fibers from wild-type and HTZ mice, Dnm2 localized in regions of intense membrane trafficking (I-band and perinuclear region), emphasizing the pathophysiological hypothesis in which DNM2-dependent trafficking would be altered. In addition, HTZ fibers showed an increased calcium concentration as well as an intracellular Dnm2 and dysferlin accumulation. A similar dysferlin retention, never reported so far in congenital myopathies, was also demonstrated in biopsies from DNM2-CNM patients and can be considered as a new marker to orientate direct genetic testing. Homozygous (HMZ) mice died during the first hours of life. Impairment of clathrin-mediated endocytosis, demonstrated in HMZ embryonic fibroblasts, could be the cause of lethality. Overall, this first mouse model of DNM2-related myopathy shows the crucial role of DNM2 in muscle homeostasis and will be a precious tool to study DNM2 functions in muscle, pathomechanisms of DNM2-CNM and developing therapeutic strategies.
常染色体显性中轴型先天性肌营养不良症(AD-CNM)是由参与胞吞作用和细胞内膜运输的 dynamin 2(DNM2)基因突变引起的。为了了解 DNM2 突变导致的病理机制,我们构建了表达最常见 AD-CNM 突变(KI-Dnm2[R465W])的基因敲入小鼠模型。杂合子(HTZ)小鼠出现了一种肌病,表现出特定的时空肌肉受累。在主要和明显受影响的胫骨前肌中,在断奶时发现收缩特性受损,并逐渐与萎缩和主要影响线粒体和网状网络的组织病理学异常相关联。在 DNM2 诱导的萎缩过程中,参与泛素-蛋白酶体和自噬途径的基因表达上调。在野生型和 HTZ 小鼠的分离肌纤维中,Dnm2 定位于强烈的膜运输区域(I 带和核周区域),强调了 DNM2 依赖性运输改变的病理生理学假说。此外,HTZ 纤维显示出钙浓度增加以及细胞内 Dnm2 和 dysferlin 积累。在 DNM2-CNM 患者的活检中也证明了从未在先天性肌营养不良中报道过的类似 dysferlin 保留,这可以被认为是指导直接基因测试的新标志物。纯合子(HMZ)小鼠在生命的头几个小时内死亡。在 HMZ 胚胎成纤维细胞中证明的网格蛋白介导的胞吞作用受损可能是导致致死性的原因。总体而言,这种首个与 DNM2 相关的肌病小鼠模型表明 DNM2 在肌肉稳态中的关键作用,并且将成为研究 DNM2 在肌肉中的功能、DNM2-CNM 的病理机制以及开发治疗策略的宝贵工具。