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

Molecular Neurodegeneration

Kristel Sleegers⁵  Christine Van Broeckhoven⁵  Marc Cruts⁵  Peter Paul De Deyn³  Rik Vandenberghe⁴  Mathieu Vandenbulcke¹  Sebastiaan Engelborghs²  Caroline Robberecht⁵  Bob Asselbergh⁵  Bavo Heeman⁵  Caroline Van Cauwenberghe⁵  Steven Vermeulen⁵  Karolien Bettens⁵ 

[1] Department of Psychiatry and Memory Clinic, University of Leuven and University Hospitals Leuven Gasthuisberg, Leuven, Belgium;Department of Neurology and Memory Clinic, Hospital Network Antwerp Middelheim and Hoge Beuken, Antwerp, Belgium;Department of Neurology and Alzheimer Research Center, University of Groningen and University Medical Center Groningen, Groningen, The Netherlands;Laboratory for Cognitive Neurology, Department of Neurology, University of Leuven and University Hospitals Leuven Gasthuisberg, Leuven, Belgium;Institute Born-Bunge, Laboratory of Neurochemistry and Behavior, University of Antwerp, Antwerp, Belgium

关键词: Golgi;    Cell secretion;    β-chain;    Rare variant;    Mutations;    Clusterin;    Alzheimer’s disease;   

Others  :  1219844
DOI  :  10.1186/s13024-015-0024-9

 received in 2015-01-27, accepted in 2015-06-30,  发布年份 2015

【 摘 要 】

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 β-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 β-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 α- and β-chain of CLU.

【 授权许可】

   
2015 Bettens et al.

【 图 表 】

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