Defining the role of autophagy in human neurodegeneration and neuronal aging

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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