Nandakumar Swapna, Vijayan Bejoy, Kishore Asha, Thekkuveettil Anoopkumar
Comprehensive Care Centre for Movement Disorders, Department of Neurology, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, 695011, India.
Division of Molecular Medicine, Department of Applied Biology, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, 695012, India.
J Cell Commun Signal. 2017 Dec;11(4):381-394. doi: 10.1007/s12079-017-0402-x. Epub 2017 Jul 26.
Increased cellular concentration of α-synuclein (α-syn) predisposes it to misfolding and aggregation that in turn impair the degradation pathways. This poses a limitation to the use of overexpression models for studies on α-syn clearance by autophagy, which is widely investigated for its therapeutic potential. This limitation can be overcome with the use of endogenous models. In this study, SK-MEL-28, a melanoma cell model with endogenous α-syn expression, was employed to study α-syn clearance through autophagy. We demonstrated the dual localization of α-syn to nucleus and cytoplasm that varied in response to changes in cellular environment. Autophagy inhibition and exposure to dopamine favored cytoplasmic localization of α-syn, while autophagy induction favored increased localization to the nucleus. The inhibitory effect of dopamine on autophagy was heightened in presence of α-syn. Additionally, because α-syn had a regulatory effect on autophagy, cells showed an increased resistance to autophagy induction in presence of α-syn. This resistance prevented effective induction of autophagy even under conditions of prolonged autophagy inhibition. These results highlight alternate physiological roles of α-syn, particularly in non-neuronal cells. Because autophagy enhancement could reverse neither the increase in α-syn levels nor the autophagy inhibition, there arises a need to evaluate the efficacy of autophagy-based therapeutic strategies.
细胞内α-突触核蛋白(α-syn)浓度的增加使其易于错误折叠和聚集,进而损害降解途径。这限制了过表达模型在通过自噬研究α-syn清除方面的应用,而自噬因其治疗潜力受到广泛研究。使用内源性模型可以克服这一限制。在本研究中,采用具有内源性α-syn表达的黑色素瘤细胞模型SK-MEL-28来研究通过自噬清除α-syn。我们证明了α-syn在细胞核和细胞质中的双重定位会随着细胞环境的变化而改变。自噬抑制和多巴胺暴露有利于α-syn在细胞质中的定位,而自噬诱导则有利于其在细胞核中的定位增加。在存在α-syn的情况下,多巴胺对自噬的抑制作用增强。此外,由于α-syn对自噬有调节作用,在存在α-syn的情况下细胞对自噬诱导的抗性增加。这种抗性即使在长期自噬抑制的条件下也能阻止自噬的有效诱导。这些结果突出了α-syn的其他生理作用,特别是在非神经元细胞中的作用。由于增强自噬既不能逆转α-syn水平的升高,也不能逆转自噬抑制,因此有必要评估基于自噬的治疗策略的疗效。