Lysosomal alterations and decreased electrophysiological activity in CLN3 disease patient-derived cortical neurons
journal contributionposted on 2023-05-21, 15:14 authored by Sueanne ChearSueanne Chear, Sharn PerrySharn Perry, Richard WilsonRichard Wilson, Aidan BindoffAidan Bindoff, Jana TalbotJana Talbot, Ware, TL, Grubman, A, James VickersJames Vickers, Pebay, A, Ruddle, JB, Anna KingAnna King, Alexander HewittAlexander Hewitt, Anthony CookAnthony Cook
CLN3 disease is a lysosomal storage disorder associated with fatal neurodegeneration that is caused by mutations in CLN3, with most affected individuals carrying at least one allele with a 966 bp deletion. Using CRISPR/Cas9, we corrected the 966 bp deletion mutation in human induced pluripotent stem cells (iPSCs) of a compound heterozygous patient (CLN3 Δ 966 bp and E295K). We differentiated these isogenic iPSCs, and iPSCs from an unrelated healthy control donor to neurons and identified disease-related changes relating to protein synthesis, trafficking and degradation, and in neuronal activity which were not apparent in CLN3- corrected or healthy control neurons. CLN3 neurons showed numerous membrane-bound vacuoles containing diverse storage material, and hyperglycosylation of the lysosomal LAMP1 protein. Proteomic analysis showed increase in lysosomal-related proteins and many ribosomal subunit proteins in CLN3 neurons that was accompanied by downregulation of proteins related to axon guidance and endocytosis. CLN3 neurons also had lower electrophysical activity as recorded using microelectrode arrays. These data implicate interrelated pathways in protein homeostasis and neurite arborization as contributing to CLN3 disease, and which could be potential targets for therapy.
Batten Disease Support and Research Association
Publication titleDisease Models & Mechanisms
Department/SchoolWicking Dementia Research Education Centre
PublisherThe Company of Biologists Ltd.
Place of publicationUnited Kingdom
Rights statement© 2022. Published by The Company of Biologists Ltd This is an Open Access article distributed under the terms of the Creative Commons Attribution International (CC BY 4.0) License, (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.