Modeling genetic risk factor of alzheimer’s disease “SORL1” using patient specific pluripotent stem cells

Abstract

Alzheimer’s disease is the most common neurodegenerative; cause of dementia and trends to increase in elder society. There is still not cure. A new strategy development for treatment is needed. SORL1 is a major genetic risk factor associated with sporadic AD. It correlates with degrading amyloid beta (Aβ) by sorting to lysosome. SORL1 overexpression has been reported that reduce Aβ production. However, the molecular mechanism that regulate in neurons is still unclear. The purpose of this study is to generate an effective drug discovery platform for identifying molecules that can modulate SORL1 level in human neurons by using induced pluripotent stem cells (iPSCs) together with genome editing using CRISPR/Cas9 technology to generate human cortical neurons SORL1-EGFP reporter cells. Here, we demonstrated that we successfully generated SORL1-EGFP reporter iPSC cell lines which still retains pluripotency and when differentiates to cortical neurons in vitro, express EGFP at the location corresponding to the location of endogenous SORL1 and SORL1-EGFP could more expressed when induced by BDNF and cAMP. Moreover, we generated the activation SORL1 using CRISPR-on technique which highly specific to endogenous. The results indicated that sgRNASORL1 could activate and significantly increase SORL1 expression levels in human erythroleukemic cell line (K562). This model could advance our understanding of SORL1 regulation, mechanisms of neurodegeneration and the effective of SORL1 activated sgRNASORL1 might offer a platform to find new therapeutics for Alzheimer’s disease in further.