Institute of Molecular Biology and Pathology (IBPM), Consiglio Nazionale delle Ricerche (CNR), c/o Sapienza University of Rome, Rome, Italy.
Unit of Cellular Networks and Molecular Therapeutic Targets, IRCCS-Regina Elena National Cancer Institute, Rome, Italy.
Eur J Neurol. 2023 Jun;30(6):1734-1744. doi: 10.1111/ene.15756. Epub 2023 Mar 26.
Microtubule defects are a common feature in several neurodegenerative disorders, including hereditary spastic paraplegia. The most frequent form of hereditary spastic paraplegia is caused by mutations in the SPG4/SPAST gene, encoding the microtubule severing enzyme spastin. To date, there is no effective therapy available but spastin-enhancing therapeutic approaches are emerging; thus prognostic and predictive biomarkers are urgently required.
An automated, simple, fast and non-invasive cell imaging-based method was developed to quantify microtubule cytoskeleton organization changes in lymphoblastoid cells and peripheral blood mononuclear cells.
It was observed that lymphoblastoid cells and peripheral blood mononuclear cells from individuals affected by SPG4-hereditary spastic paraplegia show a polarized microtubule cytoskeleton organization. In a pilot study on freshly isolated peripheral blood mononuclear cells, our method discriminates SPG4-hereditary spastic paraplegia from healthy donors and other hereditary spastic paraplegia subtypes. In addition, it is shown that our method can detect the effects of spastin protein level changes.
These findings open the possibility of a rapid, non-invasive, inexpensive test useful to recognize SPG4-hereditary spastic paraplegia subtype and evaluate the effects of spastin-enhancing drug in non-neuronal cells.
微管缺陷是几种神经退行性疾病的共同特征，包括遗传性痉挛性截瘫。最常见的遗传性痉挛性截瘫是由 SPG4/SPAST 基因突变引起的，该基因编码微管切割酶 spastin。迄今为止，尚无有效的治疗方法，但 spastin 增强治疗方法正在出现；因此迫切需要预后和预测生物标志物。
开发了一种自动化、简单、快速和非侵入性的基于细胞成像的方法，用于量化淋巴母细胞和外周血单核细胞中微管细胞骨架组织变化。
观察到患有 SPG4 遗传性痉挛性截瘫的个体的淋巴母细胞和外周血单核细胞表现出极化的微管细胞骨架组织。在对外周血单核细胞的初步研究中，我们的方法可以区分 SPG4 遗传性痉挛性截瘫与健康供体和其他遗传性痉挛性截瘫亚型。此外，还表明我们的方法可以检测 spastin 蛋白水平变化的影响。
这些发现为快速、非侵入性、廉价的测试提供了可能性，有助于识别 SPG4 遗传性痉挛性截瘫亚型并评估 spastin 增强药物在非神经元细胞中的作用。
