Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA.
Otto Loewi Research Center, Division of Pharmacology, Medical University of Graz, Austria.
Free Radic Biol Med. 2019 Sep;141:115-140. doi: 10.1016/j.freeradbiomed.2019.05.033. Epub 2019 Jun 6.
α-Synuclein (αSyn) is central to the neuropathology of Parkinson's disease (PD) due to its propensity for misfolding and aggregation into neurotoxic oligomers. Nitration/oxidation of αSyn leads to dityrosine crosslinking and aggregation. Myeloperoxidase (MPO) is an oxidant-generating enzyme implicated in neurodegenerative diseases. In the present work we have examined the impact of MPO in PD through analysis of postmortem PD brain and in a novel animal model in which we crossed a transgenic mouse expressing the human MPO (hMPO) gene to a mouse expressing human αSyn-A53T mutant (A53T) (hMPO-A53T). Surprisingly, our results show that in PD substantia nigra, the hMPO gene is expressed in neurons containing aggregates of nitrated αSyn as well as MPO-generated HOCl-modified epitopes. In our hMPO-A53T mouse model, we also saw hMPO expression in neurons but not mouse MPO. In the mouse model, hMPO was expressed in neurons colocalizing with nitrated αSyn, carbamylated lysine, nitrotyrosine, as well as HOCl-modified epitopes/proteins. RNAscope in situ hybridization confirmed hMPO mRNA expression in neurons. Interestingly, the hMPO protein expressed in hMPO-A53T brain is primarily the precursor proMPO, which enters the secretory pathway potentially resulting in interneuronal transmission of MPO and oxidative species. Importantly, the hMPO-A53T mouse model, when compared to the A53T model, exhibited significant exacerbation of motor impairment on rotating rods, balance beams, and wire hang tests. Further, hMPO expression in the A53T model resulted in earlier onset of end stage paralysis. Interestingly, there was a high concentration of αSyn aggregates in the stratum lacunosum moleculare of hippocampal CA2 region, which has been associated in humans with accumulation of αSyn pathology and neural atrophy in dementia with Lewy bodies. This accumulation of αSyn aggregates in CA2 was associated with markers of endoplasmic reticulum (ER) stress and the unfolded protein response with expression of activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), MPO, and cleaved caspase-3. Together these findings suggest that MPO plays an important role in nitrative and oxidative damage that contributes to αSyn pathology in synucleinopathies.
α-突触核蛋白(αSyn)易于错误折叠和聚集形成神经毒性寡聚物,因此是帕金森病(PD)神经病理学的核心。αSyn 的硝化/氧化导致二酪氨酸交联和聚集。在本工作中,我们通过分析 PD 死后大脑和一种新型动物模型研究了 MPO 在 PD 中的作用,该模型中我们将表达人髓过氧化物酶(MPO)基因的转基因小鼠与表达人 αSyn-A53T 突变体(A53T)的小鼠进行杂交(hMPO-A53T)。令人惊讶的是,我们的结果表明,在 PD 的黑质中,hMPO 基因在含有硝化 αSyn 聚集物的神经元中以及 MPO 生成的 HOCl 修饰表位的神经元中表达。在我们的 hMPO-A53T 小鼠模型中,我们也看到了神经元中的 hMPO 表达,但不是小鼠 MPO。在小鼠模型中,hMPO 与硝化的 αSyn、氨甲酰化赖氨酸、硝基酪氨酸以及 HOCl 修饰的表位/蛋白质共定位。RNAscope 原位杂交证实了神经元中 hMPO mRNA 的表达。有趣的是,hMPO-A53T 脑内表达的 hMPO 蛋白主要是前体 proMPO,它进入分泌途径,可能导致 MPO 和氧化物质的神经元间传递。重要的是,与 A53T 模型相比,hMPO-A53T 小鼠模型在旋转棒、平衡梁和挂线测试中表现出运动障碍的显著恶化。此外,A53T 模型中 hMPO 的表达导致终末期瘫痪的发病时间更早。有趣的是,在海马 CA2 区的腔隙分子层中有大量 αSyn 聚集物,在人类中,该区域与 αSyn 病理学的积累以及路易体痴呆症中的神经萎缩有关。这种 CA2 中 αSyn 聚集物的积累与内质网(ER)应激和未折叠蛋白反应的标志物相关,其标志物包括激活转录因子 4(ATF4)、C/EBP 同源蛋白(CHOP)、MPO 和切割的半胱天冬酶 3。这些发现表明 MPO 在硝化和氧化损伤中发挥重要作用,导致突触核蛋白病中的 αSyn 病理学。