Identification of novel regulators involved in AD pathogenesis using the CRISPR-Cas9 system

Abstract

The production of amyloid β peptide (Aβ) is an important process relating to the pathogenesis of Alzheimer disease (AD). It is widely known that the sequential cleavage of amyloid precursor protein (APP) by β- and γ-secretases lead to the production of Aβ. However, the precise regulatory mechanism for Aβ production remains unclear. We have established a CRISPR-Cas9 based screening system to identify the novel regulators of Aβ production. Calcium and integrin-binding protein 1 (CIB1) was identified as a novel potential negative regulator of Aβ production. The knockdown and knockout of Cib1 significantly increased Aβ levels. In addition, immunoprecipitation showed that CIB1 interacts with the γ-secretase complex but did not alter its enzymatic activity. Moreover, Cib1 disruption specifically reduced the cell-surface localization of the γ-secretase complex. Finally, the single-cell RNA-seq analysis in the human brain demonstrated that early-stage AD patients have lower neuronal CIB1 mRNA levels compared to healthy controls. Taken together, we have shown that CIB1 controls the subcellular localization of γ-secretase, resulting in the regulation of Aβ production, suggesting the involvement of CIB1 in the development of AD pathogenesis.