@article{f8c36d0c3f134330803264c42155b11e,
title = "Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus",
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.",
keywords = "Alzheimer's disease, Functional genomics, Haplotype variation, Nicastrin, γ-Secretase complex",
author = "{the Genetic and Environmental Risk for Alzheimer{\textquoteright}s Disease (GERAD1) Consortium} and {the Translational Genomics Research Institute (TGen) Consortium} and {the European Alzheimer Disease Initiative (EADI)} and Gillian Hamilton and Richard Killick and Lambert, {Jean Charles} and Philippe Amouyel and Carrasquillo, {Minerva M.} and Pankratz, {V. Shane} and Graff-Radford, {Neill R.} and Dickson, {Dennis W.} and Petersen, {Ronald C.} and Younkin, {Steven G.} and Powell, {John F.} and Richard Wade-Martins",
note = "Funding Information: The authors thank Dr. T. Caffrey, Dr. M. Lufino, and Dr. C. Gr{\"u}newald for helpful advice and suggestions. DNA was kindly provided by Dr. S. Harris and Professor I.J. Deary, University of Edinburgh, UK; Ncstn cell lines by Professor P. Wong, Johns Hopkins University School of Medicine, USA; the pEHHG plasmid from Professor E.A. Chiocca, Massachusetts General Hospital, USA; the Notch-CBF-1 reporter from Dr. G. Weinmaster, UCLA Medical School, USA; and the ΔEN1 plasmid from Dr. J. Aster, Brigham and Women's Hospital, USA. Brain tissue was provided by MRC, London Brain Bank for Neurodegenerative Diseases (Dr. C. Troakes), Oxford Project to Investigate Memory in Ageing (C. Joachim), and the South West Dementia Brain Bank (R. Barber). We are very grateful to the GERAD1, MAYO, TGen, and EADI consortia for sharing their data. A full list of contributors and research support for these is listed in Supplementary Figs. 1–4 . This work was supported by a Fellowship award to G.H. from the Alzheimer's Research Trust. G.H. was an Alzheimer's Society and Alzheimer Scotland Fellow. R.W.M. was a Wellcome Trust Research Career Development Fellow. Financial support for the work carried out in this manuscript was provided by Alzheimer's Research UK. ",
year = "2012",
month = aug,
day = "1",
doi = "10.1016/j.neurobiolaging.2012.02.005",
language = "English (US)",
volume = "33",
pages = "1848.e1--1848.e13",
journal = "Neurobiology of aging",
issn = "0197-4580",
publisher = "Elsevier Inc.",
number = "8",
}