James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK.
Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK.
Sci Rep. 2017 Aug 21;7(1):9003. doi: 10.1038/s41598-017-09362-3.
To examine the pathogenic role of α-synuclein (αS) in Parkinson's Disease, we have generated induced Pluripotent Stem Cell lines from early onset Parkinson's Disease patients with SNCA A53T and SNCA Triplication mutations, and in this study have differentiated them to PSC-macrophages (pMac), which recapitulate many features of their brain-resident cousins, microglia. We show that SNCA Triplication pMac, but not A53T pMac, have significantly increased intracellular αS versus controls and release significantly more αS to the medium. SNCA Triplication pMac, but not A53T pMac, show significantly reduced phagocytosis capability and this can be phenocopied by adding monomeric αS to the cell culture medium of control pMac. Fibrillar αS is taken up by pMac by actin-rearrangement-dependent pathways, and monomeric αS by actin-independent pathways. Finally, pMac degrade αS and this can be arrested by blocking lysosomal and proteasomal pathways. Together, these results show that macrophages are capable of clearing αS, but that high levels of exogenous or endogenous αS compromise this ability, likely a vicious cycle scenario faced by microglia in Parkinson's disease.
为了研究α-突触核蛋白(αS)在帕金森病中的致病作用,我们从早发性帕金森病患者中产生了具有 SNCA A53T 和 SNCA 三重复突变的诱导多能干细胞系,并在本研究中将其分化为 PSC-巨噬细胞(pMac),这些细胞再现了它们大脑常驻表亲小胶质细胞的许多特征。我们发现,与对照组相比,SNCA 三重复 pMac 而非 A53T pMac 具有显著增加的细胞内αS,并且向培养基中释放更多的αS。与 A53T pMac 不同,SNCA 三重复 pMac 显示出显著降低的吞噬能力,而将单体αS 添加到对照 pMac 的细胞培养基中可以模拟这种情况。纤维状αS 通过肌动蛋白重排依赖途径被 pMac 摄取,而单体αS 通过肌动蛋白非依赖途径摄取。最后,pMac 降解αS,并且可以通过阻断溶酶体和蛋白酶体途径来阻止其降解。总之,这些结果表明巨噬细胞能够清除αS,但高水平的外源性或内源性αS 损害了这种能力,这可能是帕金森病中小胶质细胞面临的恶性循环情况。