Liu Yanying, Qiao Fangfang, Leiferman Patricia C, Ross Alan, Schlenker Evelyn H, Wang Hongmin
Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, Vermillion, SD 57069, USA.
Sanford Medical Genetics Laboratory, Sioux Falls, SD 57105, USA.
Hum Mol Genet. 2017 Nov 15;26(22):4416-4428. doi: 10.1093/hmg/ddx327.
Although it has been speculated that proteasome dysfunction may contribute to the pathogenesis of Huntington's disease (HD), a devastating neurodegenerative disorder, how proteasome activity is regulated in HD affected stem cells and somatic cells remains largely unclear. To better understand the pathogenesis of HD, we analyzed proteasome activity and the expression of FOXO transcription factors in three wild-type (WT) and three HD induced-pluripotent stem cell (iPSC) lines. HD iPSCs exhibited elevated proteasome activity and higher levels of FOXO1 and FOXO4 proteins. Knockdown of FOXO4 but not FOXO1 expression decreased proteasome activity. Following neural differentiation, the HD-iPSC-derived neural progenitor cells (NPCs) demonstrated lower levels of proteasome activity and FOXO expressions than their WT counterparts. More importantly, overexpression of FOXO4 but not FOXO1 in HD NPCs dramatically enhanced proteasome activity. When HD NPCs were further differentiated into DARPP32-positive neurons, these HD neurons were more susceptible to death than WT neurons and formed Htt aggregates under the condition of oxidative stress. Similar to HD NPCs, HD-iPSC-derived neurons showed reduced proteasome activity and diminished FOXO4 expression compared to WT-iPSC-derived neurons. Furthermore, HD iPSCs had lower AKT activities than WT iPSCs, whereas the neurons derived from HD iPSC had higher AKT activities than their WT counterparts. Inhibiting AKT activity increased both FOXO4 level and proteasome activity, indicating a potential role of AKT in regulating FOXO levels. These data suggest that FOXOs modulate proteasome activity, and thus represents a potentially valuable therapeutic target for HD.
尽管有人推测蛋白酶体功能障碍可能与亨廷顿舞蹈病(HD)的发病机制有关,HD是一种毁灭性的神经退行性疾病,但在HD影响的干细胞和体细胞中蛋白酶体活性是如何调节的仍不清楚。为了更好地理解HD的发病机制,我们分析了三个野生型(WT)和三个HD诱导多能干细胞(iPSC)系中的蛋白酶体活性以及FOXO转录因子的表达。HD iPSC表现出蛋白酶体活性升高以及FOXO1和FOXO4蛋白水平更高。敲低FOXO4而非FOXO1的表达会降低蛋白酶体活性。神经分化后,HD-iPSC来源的神经祖细胞(NPC)的蛋白酶体活性和FOXO表达水平低于其WT对应细胞。更重要的是,在HD NPC中过表达FOXO4而非FOXO1会显著增强蛋白酶体活性。当HD NPC进一步分化为DARPP32阳性神经元时,这些HD神经元比WT神经元更容易死亡,并且在氧化应激条件下形成Htt聚集体。与HD NPC相似,与WT-iPSC来源的神经元相比,HD-iPSC来源的神经元蛋白酶体活性降低且FOXO4表达减少。此外,HD iPSC的AKT活性低于WT iPSC,而HD iPSC来源的神经元的AKT活性高于其WT对应细胞。抑制AKT活性会增加FOXO4水平和蛋白酶体活性,表明AKT在调节FOXO水平中具有潜在作用。这些数据表明FOXO调节蛋白酶体活性,因此是HD潜在的有价值的治疗靶点。