Macken William L, Vandrovcova Jana, Hanna Michael G, Pitceathly Robert D S
Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology and The National Hospital for Neurology and Neurosurgery, London, UK.
Nat Rev Neurol. 2021 Apr;17(4):215-230. doi: 10.1038/s41582-021-00455-2. Epub 2021 Feb 23.
Next-generation sequencing (NGS) has increased our understanding of the molecular basis of many primary mitochondrial diseases (PMDs). Despite this progress, many patients with suspected PMD remain without a genetic diagnosis, which restricts their access to in-depth genetic counselling, reproductive options and clinical trials, in addition to hampering efforts to understand the underlying disease mechanisms. Although they represent a considerable improvement over their predecessors, current methods for sequencing the mitochondrial and nuclear genomes have important limitations, and molecular diagnostic techniques are often manual and time consuming. However, recent advances in genomics and transcriptomics offer realistic solutions to these challenges. In this Review, we discuss the current genetic testing approach for PMDs and the opportunities that exist for increased use of whole-genome NGS of nuclear and mitochondrial DNA (mtDNA) in the clinical environment. We consider the possible role for long-read approaches in sequencing of mtDNA and in the identification of novel nuclear genomic causes of PMDs. We examine the expanding applications of RNA sequencing, including the detection of cryptic variants that affect splicing and gene expression and the interpretation of rare and novel mitochondrial transfer RNA variants.
新一代测序(NGS)增进了我们对许多原发性线粒体疾病(PMD)分子基础的理解。尽管取得了这一进展,但许多疑似PMD患者仍未得到基因诊断,这不仅限制了他们获得深入基因咨询、生殖选择和临床试验的机会,还阻碍了对潜在疾病机制的研究。尽管目前用于线粒体和核基因组测序的方法相较于之前有了显著改进,但仍存在重要局限性,而且分子诊断技术往往需要人工操作且耗时。然而,基因组学和转录组学的最新进展为这些挑战提供了切实可行的解决方案。在本综述中,我们讨论了目前针对PMD的基因检测方法,以及在临床环境中增加使用核DNA和线粒体DNA(mtDNA)全基因组NGS的机会。我们考虑了长读长方法在mtDNA测序以及识别PMD新的核基因组病因方面可能发挥的作用。我们研究了RNA测序不断扩展的应用,包括检测影响剪接和基因表达的隐匿变异以及解读罕见和新出现的线粒体转运RNA变异。