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A recurrent gain-of-function mutation in CLCN6, encoding the ClC-6 Cl-/H+-exchanger, causes early-onset neurodegeneration

Item Type:Article
Title:A recurrent gain-of-function mutation in CLCN6, encoding the ClC-6 Cl-/H+-exchanger, causes early-onset neurodegeneration
Creators Name:Polovitskaya, M.M. and Barbini, C. and Martinelli, D. and Harms, F.L. and Cole, F.S. and Calligari, P. and Bocchinfuso, G. and Stella, L. and Ciolfi, A. and Niceta, M. and Rizza, T. and Shinawi, M. and Sisco, K. and Johannsen, J. and Denecke, J. and Carrozzo, R. and Wegner, D.J. and Kutsche, K. and Tartaglia, M. and Jentsch, T.J.
Abstract:Dysfunction of the endolysosomal system is often associated with neurodegenerative disease because postmitotic neurons are particularly reliant on the elimination of intracellular aggregates. Adequate function of endosomes and lysosomes requires finely tuned luminal ion homeostasis and transmembrane ion fluxes. Endolysosomal CLC Cl(-)/H(+) exchangers function as electric shunts for proton pumping and in luminal Cl(-) accumulation. We now report three unrelated children with severe neurodegenerative disease, who carry the same de novo c.1658A>G (p.Tyr553Cys) mutation in CLCN6, encoding the late endosomal Cl(-)/H(+)-exchanger ClC-6. Whereas Clcn6(-/-) mice have only mild neuronal lysosomal storage abnormalities, the affected individuals displayed severe developmental delay with pronounced generalized hypotonia, respiratory insufficiency, and variable neurodegeneration and diffusion restriction in cerebral peduncles, midbrain, and/or brainstem in MRI scans. The p.Tyr553Cys amino acid substitution strongly slowed ClC-6 gating and increased current amplitudes, particularly at the acidic pH of late endosomes. Transfection of ClC-6(Tyr553Cys), but not ClC-6(WT), generated giant LAMP1-positive vacuoles that were poorly acidified. Their generation strictly required ClC-6 ion transport, as shown by transport-deficient double mutants, and depended on Cl(-)/H(+) exchange, as revealed by combination with the uncoupling p.Glu200Ala substitution. Transfection of either ClC-6(Tyr553Cys/Glu200Ala) or ClC-6(Glu200Ala) generated slightly enlarged vesicles, suggesting that p.Glu200Ala, previously associated with infantile spasms and microcephaly, is also pathogenic. Bafilomycin treatment abrogated vacuole generation, indicating that H(+)-driven Cl(-) accumulation osmotically drives vesicle enlargement. Our work establishes mutations in CLCN6 associated with neurological diseases, whose spectrum of clinical features depends on the differential impact of the allele on ClC-6 function.
Keywords:Channelopathy, Anion/Proton Antiport, Chloride/Proton Exchange, Chloride Channel, Neurogenic Bladder, Vacuole Fusion, Gain of Function, Gating Glutamate, Luminal pH, Copper Metabolism, Animals, Cricetulus, Mice
Source:American Journal of Human Genetics
ISSN:0002-9297
Publisher:Cell Press
Volume:107
Number:6
Page Range:1062-1077
Date:3 December 2020
Official Publication:https://doi.org/10.1016/j.ajhg.2020.11.004
PubMed:View item in PubMed

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