Division of Medical Genetics and Genomics, The Children's Hospital, Zhejiang University School of Medicine / National Clinical Research Center for Child Health, Hangzhou 310052, China.
Institute of Genetics and Department of Human Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
J Zhejiang Univ Sci B. 2020;21(11):885-896. doi: 10.1631/jzus.B2000339.
The pathogenesis of very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is highly heterogeneous and still unclear. Additional novel variants have been recently detected in the population. The molecular and cellular effects of these previously unreported variants are still poorly understood and require further characterization. To address this problem, we have evaluated the various functions and biochemical consequences of six novel missense variants that lead to mild VLCAD deficiency. Marked deficiencies in fatty acid oxidation (FAO) and other mitochondrial defects were observed in cells carrying one of these six variants (c.541C>T, c.863T>G, c.895A>G, c.1238T>C, c.1276G>A, and c.1505T>A), including reductions in mitochondrial respiratory-chain function and adenosine triphosphate (ATP) production, and increased levels of mitochondrial reactive oxygen species (ROS). Intriguingly, higher apoptosis levels were found in cells carrying the mutant VLCAD under glucose-limited stress. Moreover, the stability of the mutant homodimer was disturbed, and major conformational changes in each mutant VLCAD structure were predicted by molecular dynamics (MD) simulation. The data presented here may provide valuable information for improving management of diagnosis and treatment of VLCAD deficiency and for a better understanding of the general molecular bases of disease variability.
极长链酰基辅酶 A 脱氢酶 (VLCAD) 缺乏症的发病机制高度异质,目前仍不清楚。最近在人群中发现了其他新的变异体。这些以前未报道的变异体的分子和细胞效应仍知之甚少,需要进一步表征。为了解决这个问题,我们评估了导致轻度 VLCAD 缺乏症的六个新错义变异体的各种功能和生化后果。在携带其中一个变体(c.541C>T、c.863T>G、c.895A>G、c.1238T>C、c.1276G>A 和 c.1505T>A)的细胞中观察到脂肪酸氧化 (FAO) 和其他线粒体缺陷的明显缺乏,包括线粒体呼吸链功能和三磷酸腺苷 (ATP) 产生减少,以及线粒体活性氧 (ROS) 水平增加。有趣的是,在葡萄糖限制应激下携带突变 VLCAD 的细胞中发现了更高的细胞凋亡水平。此外,突变体同源二聚体的稳定性受到干扰,并且通过分子动力学 (MD) 模拟预测了每个突变 VLCAD 结构的主要构象变化。这里呈现的数据可能为改善 VLCAD 缺乏症的诊断和治疗管理提供有价值的信息,并为更好地了解疾病变异性的一般分子基础提供信息。