Fuller Heidi R, Mandefro Berhan, Shirran Sally L, Gross Andrew R, Kaus Anjoscha S, Botting Catherine H, Morris Glenn E, Sareen Dhruv
Wolfson Centre for Inherited Neuromuscular Disease, The Robert Jones and Agnes Hunt Orthopaedic HospitalOswestry, UK; Institute for Science and Technology in Medicine, Keele UniversityStaffordshire, UK.
Board of Governors-Regenerative Medicine Institute, Cedars-Sinai Medical CenterLos Angeles, CA, USA; iPSC Core, The David and Janet Polak Foundation Stem Cell Core LaboratoryLos Angeles, CA, USA.
Front Cell Neurosci. 2016 Jan 11;9:506. doi: 10.3389/fncel.2015.00506. eCollection 2015.
Spinal muscular atrophy (SMA) is an inherited neuromuscular disease primarily characterized by degeneration of spinal motor neurons, and caused by reduced levels of the SMN protein. Previous studies to understand the proteomic consequences of reduced SMN have mostly utilized patient fibroblasts and animal models. We have derived human motor neurons from type I SMA and healthy controls by creating their induced pluripotent stem cells (iPSCs). Quantitative mass spectrometry of these cells revealed increased expression of 63 proteins in control motor neurons compared to respective fibroblasts, whereas 30 proteins were increased in SMA motor neurons vs. their fibroblasts. Notably, UBA1 was significantly decreased in SMA motor neurons, supporting evidence for ubiquitin pathway defects. Subcellular distribution of UBA1 was predominantly cytoplasmic in SMA motor neurons in contrast to nuclear in control motor neurons; suggestive of neurodevelopmental abnormalities. Many of the proteins that were decreased in SMA motor neurons, including beta III-tubulin and UCHL1, were associated with neurodevelopment and differentiation. These neuron-specific consequences of SMN depletion were not evident in fibroblasts, highlighting the importance of iPSC technology. The proteomic profiles identified here provide a useful resource to explore the molecular consequences of reduced SMN in motor neurons, and for the identification of novel biomarker and therapeutic targets for SMA.
脊髓性肌萎缩症(SMA)是一种遗传性神经肌肉疾病,主要特征为脊髓运动神经元变性,由运动神经元存活蛋白(SMN)水平降低所致。以往旨在了解SMN水平降低所产生蛋白质组学影响的研究大多使用患者成纤维细胞和动物模型。我们通过诱导多能干细胞(iPSC)技术,从I型SMA患者和健康对照者中获得了人类运动神经元。对这些细胞进行的定量质谱分析显示,与各自的成纤维细胞相比,对照运动神经元中有63种蛋白质表达增加,而SMA运动神经元与其成纤维细胞相比,有30种蛋白质表达增加。值得注意的是,SMA运动神经元中UBA1显著减少,这为泛素途径缺陷提供了证据支持。与对照运动神经元中UBA1主要分布于细胞核不同,SMA运动神经元中UBA1主要分布于细胞质,提示存在神经发育异常。SMA运动神经元中许多减少的蛋白质,包括βIII微管蛋白和泛素羧基末端水解酶L1(UCHL1),都与神经发育和分化相关。在成纤维细胞中未发现SMN缺失的这些神经元特异性影响,突出了iPSC技术的重要性。本文确定的蛋白质组学图谱为探索运动神经元中SMN水平降低所产生分子影响以及识别SMA的新型生物标志物和治疗靶点提供了有用资源。