Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus.

Gillian Hamilton, Richard Killick, [No Value] Genetic, Genomics Research Institute Consortium Translational Genomics Research Institute Consortium, Jean Charles Lambert, Philippe Amouyel, Alzheimer Disease Initiative European Alzheimer Disease Initiative, Minerva M. Carrasquillo, V. Shane Pankratz, Neill R. Graff-Radford, Dennis W. Dickson, Ronald C. Petersen, Steven G. Younkin, John F. Powell, Richard Wade-Martins

Research output: Contribution to journalArticle

Abstract

Nicastrin (NCSTN) is a component of the γ-secretase complex and therefore potentially a candidate risk gene for Alzheimer's disease. Here, we have developed a novel functional genomics methodology to express common locus haplotypes to assess functional differences. DNA recombination was used to engineer 5 bacterial artificial chromosomes (BACs) to each express a different haplotype of the NCSTN locus. Each NCSTN-BAC was delivered to knockout nicastrin (Ncstn(-/-)) cells and clonal NCSTN-BAC(+)/Ncstn(-/-) cell lines were created for functional analyses. We showed that all NCSTN-BAC haplotypes expressed nicastrin protein and rescued γ-secretase activity and amyloid beta (Aβ) production in NCSTN-BAC(+)/Ncstn(-/-) lines. We then showed that genetic variation at the NCSTN locus affected alternative splicing in human postmortem brain tissue. However, there was no robust functional difference between clonal cell lines rescued by each of the 5 different haplotypes. Finally, there was no statistically significant association of NCSTN with disease risk in the 4 cohorts. We therefore conclude that it is unlikely that common variation at the NCSTN locus is a risk factor for Alzheimer's disease.

Original languageEnglish (US)
JournalNeurobiology of Aging
Volume33
Issue number8
StatePublished - Aug 2012
Externally publishedYes

Fingerprint

Bacterial Artificial Chromosomes
Sequence Analysis
Haplotypes
Amyloid Precursor Protein Secretases
Alzheimer Disease
Cell Line
Alternative Splicing
Genomics
Amyloid
Genetic Recombination
DNA
Brain
Genes

ASJC Scopus subject areas

  • Clinical Neurology
  • Neuroscience(all)
  • Aging
  • Developmental Biology
  • Geriatrics and Gerontology

Cite this

Hamilton, G., Killick, R., Genetic, N. V., Translational Genomics Research Institute Consortium, G. R. I. C., Lambert, J. C., Amouyel, P., ... Wade-Martins, R. (2012). Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus. Neurobiology of Aging, 33(8).

Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus. / Hamilton, Gillian; Killick, Richard; Genetic, [No Value]; Translational Genomics Research Institute Consortium, Genomics Research Institute Consortium; Lambert, Jean Charles; Amouyel, Philippe; European Alzheimer Disease Initiative, Alzheimer Disease Initiative; Carrasquillo, Minerva M.; Pankratz, V. Shane; Graff-Radford, Neill R.; Dickson, Dennis W.; Petersen, Ronald C.; Younkin, Steven G.; Powell, John F.; Wade-Martins, Richard.

In: Neurobiology of Aging, Vol. 33, No. 8, 08.2012.

Research output: Contribution to journalArticle

Hamilton, G, Killick, R, Genetic, NV, Translational Genomics Research Institute Consortium, GRIC, Lambert, JC, Amouyel, P, European Alzheimer Disease Initiative, ADI, Carrasquillo, MM, Pankratz, VS, Graff-Radford, NR, Dickson, DW, Petersen, RC, Younkin, SG, Powell, JF & Wade-Martins, R 2012, 'Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus.', Neurobiology of Aging, vol. 33, no. 8.
Hamilton G, Killick R, Genetic NV, Translational Genomics Research Institute Consortium GRIC, Lambert JC, Amouyel P et al. Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus. Neurobiology of Aging. 2012 Aug;33(8).
Hamilton, Gillian ; Killick, Richard ; Genetic, [No Value] ; Translational Genomics Research Institute Consortium, Genomics Research Institute Consortium ; Lambert, Jean Charles ; Amouyel, Philippe ; European Alzheimer Disease Initiative, Alzheimer Disease Initiative ; Carrasquillo, Minerva M. ; Pankratz, V. Shane ; Graff-Radford, Neill R. ; Dickson, Dennis W. ; Petersen, Ronald C. ; Younkin, Steven G. ; Powell, John F. ; Wade-Martins, Richard. / Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus. In: Neurobiology of Aging. 2012 ; Vol. 33, No. 8.
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AU - Lambert, Jean Charles

AU - Amouyel, Philippe

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