A Large Panel of Isogenic APP and PSEN1 Mutant Human iPSC Neurons Reveals Shared Endosomal Abnormalities Mediated by APP β-CTFs, Not Aβ

Dylan Kwart; Andrew Gregg; Claudia Scheckel; Elisabeth A Murphy; Dominik Paquet; Michael Duffield; John Fak; Olav Olsen; Robert B Darnell; Marc Tessier-Lavigne

doi:10.1016/j.neuron.2019.07.010

A Large Panel of Isogenic APP and PSEN1 Mutant Human iPSC Neurons Reveals Shared Endosomal Abnormalities Mediated by APP β-CTFs, Not Aβ

Neuron. 2019 Oct 23;104(2):256-270.e5. doi: 10.1016/j.neuron.2019.07.010. Epub 2019 Aug 12.

Authors

Affiliations

¹ Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
² Institute of Neuropathology, University of Zurich, Schmelzbergstrasse 12, 8091 Zurich, Switzerland.
³ Laboratory of Neuro-oncology and Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA.
⁴ Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA; Institute for Stroke and Dementia Research (ISD), University Hospital, LMU Munich, 81377 Munich, Germany.
⁵ Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA; Department of Biology, Stanford University, Stanford, CA 94305, USA. Electronic address: tessier3@stanford.edu.

PMID: 31416668
DOI: 10.1016/j.neuron.2019.07.010

Abstract

Familial Alzheimer's disease (fAD) results from mutations in the amyloid precursor protein (APP) and presenilin (PSEN1 and PSEN2) genes. Here we leveraged recent advances in induced pluripotent stem cell (iPSC) and CRISPR/Cas9 genome editing technologies to generate a panel of isogenic knockin human iPSC lines carrying APP and/or PSEN1 mutations. Global transcriptomic and translatomic profiling revealed that fAD mutations have overlapping effects on the expression of AD-related and endocytosis-associated genes. Mutant neurons also increased Rab5+ early endosome size. APP and PSEN1 mutations had discordant effects on Aβ production but similar effects on APP β C-terminal fragments (β-CTFs), which accumulate in all mutant neurons. Importantly, endosomal dysfunction correlated with accumulation of β-CTFs, not Aβ, and could be rescued by pharmacological modulation of β-secretase (BACE). These data display the utility of our mutant iPSCs in studying AD-related phenotypes in a non-overexpression human-based system and support mounting evidence that β-CTF may be critical in AD pathogenesis.

Keywords: Alzheimer’s disease; Aβ; CRISPR/Cas9; Rab5; amyloid precursor protein; amyloid-beta; beta-C-terminal fragment; endocytosis; induced pluripotent stem cell; presenilin; β-CTF.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Alzheimer Disease / genetics*
Alzheimer Disease / pathology
Amyloid Precursor Protein Secretases
Amyloid beta-Peptides / metabolism*
Amyloid beta-Protein Precursor / genetics*
Aspartic Acid Endopeptidases
CRISPR-Cas Systems
Cell Line
Endocytosis / genetics*
Endosomes / metabolism*
Endosomes / pathology
Gene Expression Profiling
Gene Knock-In Techniques
Heterozygote
Homozygote
Humans
Induced Pluripotent Stem Cells
Mutation
Neurons / metabolism*
Organelle Size
Peptide Fragments / metabolism*
Phenotype
Presenilin-1 / genetics*
Proteomics
rab5 GTP-Binding Proteins / metabolism

Substances

APP protein, human
Amyloid beta-Peptides
Amyloid beta-Protein Precursor
PSEN1 protein, human
Peptide Fragments
Presenilin-1
amyloid beta-protein (1-42)
Amyloid Precursor Protein Secretases
Aspartic Acid Endopeptidases
BACE1 protein, human
RAB5C protein, human
rab5 GTP-Binding Proteins