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P3‐166: SUBCELLULAR CO‐LOCALIZATION OF APP AND APP‐CLEAVING ENZYMES
Author(s) -
Satir Tugce Munise,
Agholme Lotta,
Nazir Faisal Hayat,
Bergström Petra,
Zetterberg Henrik
Publication year - 2018
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2018.06.1524
Subject(s) - amyloid precursor protein , microbiology and biotechnology , endosome , chemistry , immunocytochemistry , neurogenesis , intracellular , biology , alzheimer's disease , medicine , disease , endocrinology
AD-associated variants in SORL1 lead to detectable and modifiable endocytic phenotypes in human neurons. Methods: hiPSC-derived neurons are generated from control and patient fibroblasts. Neurons are generated from hiPSCs using established differentiation protocols. Patient hiPSC lines harbor predicted pathogenic variants in SORL1. CRISPR/Cas9 will be used to correct SORL1 variants. Analysis of endosomal network dysfunction and cellular AD phenotypes will be: Rab5 immunostaining, Transferring recycling assay, amyloid beta ELISA, phospho-Tau ELISA. Results: In hiPSC-derived neural cells expressing a SORL1 shRNA, we show increased amyloid beta peptides, reduced transferring recycling, and increased size of Rab5+ endosomes. In collaboration with the UWAlzheimer’s Disease Research Center, we have identified AD patients with SORL1 coding variants and are generating hiPSCs from these patients. We will assay hiPSC-derived neurons from SORL1 variant carriers for endosomal phenotypes and use CRISPR/Cas9 gene-editing to correct the variants to determine whether the predicted pathogenic variant leads to endocytic dysfunction in human neurons.Conclusions:This work is significant in that it will investigate a functional genotype-phenotype relationship of genetic variants in the endosomal network, which is known to be disrupted early in AD pathogenesis. Investigating this driver of disease pathogenesis and how it relates to human genetic background is critical in the development of new and precision treatments for AD.