期刊
DISEASE MODELS & MECHANISMS
卷 15, 期 12, 页码 -出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/dmm.049651
关键词
Juvenile neuronal ceroid lipofuscinosis; Batten disease; Dementia; Induced pluripotent stem cells; Microelectrode array; Proteomics
资金
- Batten Disease Support and Research Association Australia [RGP204]
- Royal Hobart Hospital Research Foundation [17-205]
- National Health and Medical Research Council Dementia Leadership Fellowship [APP1136913]
- Batten Disease Support and Research Association Australia
CLN3 disease is a fatal neurodegenerative disorder caused by mutations in the CLN3 gene. We corrected the mutation in human induced pluripotent stem cells (iPSCs) using CRISPR/Cas9 technology and observed disease-related changes in protein synthesis, trafficking and degradation, and neuronal activity.
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 Delta 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, 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 inter-related pathways in protein homeostasis and neurite arborization as contributing to CLN3 disease, and which could be potential targets for therapy.
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