Reduced secreted clusterin as a mechanism for Alzheimer-associated CLU mutations

Karolien Bettens, Steven Vermeulen, Caroline Van Cauwenberghe, Bavo Heeman, Bob Asselbergh, Caroline Robberecht, Sebastiaan Engelborghs, Mathieu Vandenbulcke, Rik Vandenberghe, Peter Paul De Deyn, Marc Cruts, Christine Van Broeckhoven*, Kristel Sleegers

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Background: The clusterin (CLU) gene has been identified as an important risk locus for Alzheimer's disease (AD). Although the actual risk-increasing polymorphisms at this locus remain to be identified, we previously observed an increased frequency of rare non-synonymous mutations and small insertion-deletions of CLU in AD patients, which specifically clustered in the beta-chain domain of CLU. Nonetheless the pathogenic nature of these variants remained unclear. Here we report a novel non-synonymous CLU mutation (p.I360N) in a Belgian Alzheimer patient and have explored the pathogenic nature of this and 10 additional CLU mutations on protein localization and secretion in vitro using immunocytochemistry, immunodetection and ELISAs.

Results: Three patient-specific CLU mutations in the beta-chain (p.I303NfsX13, p.R338W and p.I360N) caused an alteration of the subcellular CLU localization and diminished CLU transport through the secretory pathway, indicative of possible degradation mechanisms. For these mutations, significantly reduced CLU intensity was observed in the Golgi while almost all CLU protein was exclusively present in the endoplasmic reticulum. This was further confirmed by diminished CLU secretion in HEK293T and HEK293 FLp-In cell lines.

Conclusions: Our data lend further support to the contribution of rare coding CLU mutations in the pathogenesis of neurodegenerative diseases. Functional analyses suggest reduced secretion of the CLU protein as the mode of action for three of the examined CLU mutations. One of those is a frameshift mutation leading to a loss of secreted protein, and the other two mutations are amino acid substitutions in the disulfide bridge region, possibly interfering with heterodimerization of the alpha- and beta-chain of CLU.

Original languageEnglish
Article number30
Number of pages12
JournalMolecular Neurodegeneration
Volume10
DOIs
Publication statusPublished - 16-Jul-2015

Keywords

  • Alzheimer's disease
  • Clusterin
  • Mutations
  • Rare variant
  • beta-chain
  • Cell secretion
  • Golgi
  • GENOME-WIDE ASSOCIATION
  • IDENTIFIES VARIANTS
  • EXPRESSION
  • DISEASE
  • PROTEIN
  • DEMENTIA
  • BRAIN

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