Defining the role of autophagy in human neurodegeneration and neuronal aging

基于动物和人类癌症干细胞系的模型的主要缺点之一是，当应用于人类中枢神经系统时，预测疾病相关结果的成功程度有限。因此，在相关的受疾病影响的人类神经元亚型中，有一个巨大的需求来定义直接的机制见解。自噬是神经发生、神经退行性变和神经元老化的重要过程。我的研究将以人类多能干细胞（hPSC）衍生的神经元为模型系统，追求以下目标：i)我将产生基因工程的hPSC细胞系，在神经发育和成熟的任何给定时间点都能严格控制自噬通量。ii)我将分析抑制自噬对确定身份的纯神经元中蛋白质聚集的影响，并通过细胞分离和随后的silac -质谱法建立每种神经元亚型特异性聚集蛋白质组图谱。iii)使用progerin诱导的神经元衰老作为特定衰老标志的参考，我将评估自噬抑制促进人类神经元衰老相关改变的能力，以及自噬诱导逆转这些自噬的能力，以努力逆转衰老标志，并作为一种可能的年轻化策略。提出的方法将使研究人类神经发生，特定身份的成熟神经元中蛋白质聚集机制以及自噬对神经元衰老细胞标志的贡献成为可能。我希望该项目将增加对自噬如何促进人类神经发生的重要知识，以及它如何影响神经元亚型特异性蛋白质聚集谱，以及自噬的下降如何促进神经元衰老的标志。最后，我预计这些结果将对我们理解人类衰老过程中健康的神经元稳态做出有价值的贡献。并指出神经退行性疾病的候选治疗途径

One of the major shortcomings of animal and human cancer stem cell line-based models is the limited success in predicting outcomedisease-relevant outcomes when applied to the human CNS. Accordingly, there is a huge need for defining mechanistic insights directly in the relevant human neuronsneuron subtypes affected by disease. This is especially the case infor the fieldprocess of autophagy, a processwhich is important for neurogenesis, neurodegeneration and neuronal aging. My studies will pursue the following aims using human pluripotent stem cell (hPSC)-derived neurons as model system:i) I will generate genetically engineered hPSCs lines that allow a tight control of autophagic flux at any given time point of neural development and maturation. ii) I will analyze the effect of inhibiting autophagy on protein aggregation in pure neurons of defined identity and establish profiles of the aggregating proteome specific for each neuronal subtype by cell fractionation and subsequent SILAC-mass spectrometry.iii) Using progerin-induced neuronal aging as a reference for specific aging hallmarks, I will assess the capacity of autophagy inhibition on promoting aging-related alterations in human neurons as well as autophagythe induction for the reversion of thoseautophagy in an effort to reverse aging hallmarks and as a possible rejuvenation strategy.The proposed approach will enable the study of human neurogenesis, mechanisms of protein aggregation in mature neurons of specific identity and the contribution of autophagy on the cellular hallmarks of neuronal aging. I expect that the project will add significant knowledge on how autophagy contributes to human neurogenesis and, how it impacts neuron subtype-specific protein aggregation profiles, and on how a decline in autophagy may promote hallmarks of neuronal aging. Finally, I anticipate that the results will make a valuable contribution to our understanding of healthy neuronal homeostasis during human aging. and point to candidate therapeutic pathways in neurodegenerative disease