Joshi-Barr Shivanjali, Bett Cyrus, Chiang Wei-Chieh, Trejo Margarita, Goebel Hans H, Sikorska Beata, Liberski Pawel, Raeber Alex, Lin Jonathan H, Masliah Eliezer, Sigurdson Christina J
Department of Pathology, School of Medicine, University of California, San Diego, La Jolla, California, USA.
J Virol. 2014 Feb;88(4):2071-82. doi: 10.1128/JVI.02279-13. Epub 2013 Dec 4.
In certain sporadic, familial, and infectious prion diseases, the prion protein misfolds and aggregates in skeletal muscle in addition to the brain and spinal cord. In myocytes, prion aggregates accumulate intracellularly, yet little is known about clearance pathways. Here we investigated the clearance of prion aggregates in muscle of transgenic mice that develop prion disease de novo. In addition to neurodegeneration, aged mice developed a degenerative myopathy, with scattered myocytes containing ubiquitinated, intracellular prion inclusions that were adjacent to myocytes lacking inclusions. Myocytes also showed elevated levels of the endoplasmic reticulum chaperone Grp78/BiP, suggestive of impaired protein degradation and endoplasmic reticulum stress. Additionally, autophagy was induced, as indicated by increased levels of beclin-1 and LC3-II. In C2C12 myoblasts, inhibition of autophagosome maturation or lysosomal degradation led to enhanced prion aggregation, consistent with a role for autophagy in prion aggregate clearance. Taken together, these findings suggest that the induction of autophagy may be a central strategy for prion aggregate clearance in myocytes. IMPORTANCE In prion diseases, the prion protein misfolds and aggregates in the central nervous system and sometimes in other organs, including muscle, yet the cellular pathways of prion aggregate clearance are unclear. Here we investigated the clearance of prion aggregates in the muscle of a transgenic mouse model that develops profound muscle degeneration. We found that endoplasmic reticulum stress pathways were activated and that autophagy was induced. Blocking of autophagic degradation in cell culture models led to an accumulation of aggregated prion protein. Collectively, these findings suggest that autophagy has an instrumental role in prion protein clearance.
在某些散发性、家族性和传染性朊病毒疾病中,朊病毒蛋白除了在脑和脊髓中发生错误折叠并聚集外,在骨骼肌中也会如此。在肌细胞中,朊病毒聚集体在细胞内积累,但关于其清除途径却知之甚少。在此,我们研究了从头发生朊病毒疾病的转基因小鼠肌肉中朊病毒聚集体的清除情况。除了神经退行性变外,老龄小鼠还出现了退行性肌病,散在的肌细胞含有泛素化的细胞内朊病毒包涵体,这些包涵体与缺乏包涵体的肌细胞相邻。肌细胞还显示内质网伴侣蛋白Grp78/BiP水平升高,提示蛋白质降解受损和内质网应激。此外,自噬被诱导,这可通过beclin-1和LC3-II水平的升高得以表明。在C2C12成肌细胞中,自噬体成熟或溶酶体降解的抑制导致朊病毒聚集增强,这与自噬在朊病毒聚集体清除中的作用一致。综上所述,这些发现表明自噬的诱导可能是肌细胞中朊病毒聚集体清除的核心策略。重要性:在朊病毒疾病中,朊病毒蛋白在中枢神经系统以及有时在包括肌肉在内的其他器官中发生错误折叠并聚集,然而朊病毒聚集体清除的细胞途径尚不清楚。在此,我们研究了一个发生严重肌肉退化的转基因小鼠模型肌肉中朊病毒聚集体的清除情况。我们发现内质网应激途径被激活且自噬被诱导。在细胞培养模型中阻断自噬降解导致聚集的朊病毒蛋白积累。总体而言,这些发现表明自噬在朊病毒蛋白清除中具有重要作用。