Quantitation of heteroplasmy of mtDNA sequence variants identified in a population of AD patients and controls by array-based resequencing

被引:39
作者
Coon, Keith D.
Valla, Jon
Szelinger, Szabolics
Schneider, Lonnie E.
Niedzielko, Tracy L.
Brown, Kevin M.
Pearson, John V.
Halperin, Rebecca
Dunckley, Travis
Papassotiropoulos, Andreas
Caselli, Richard J.
Reiman, Eric M.
Stephan, Dietrich A. [1 ]
机构
[1] Translat Genom Res Inst, Neurogenom Div, Phoenix, AZ 85004 USA
[2] St Josephs Hosp, Barrow Neurol Inst, Phoenix, AZ 85013 USA
[3] Univ Zurich, Div Psychiat Res, Zurich, Switzerland
[4] Mayo Clin, Dept Neurol, Scottsdale, AZ USA
[5] Univ Arizona, Dept Psychiat, PET Ctr, Banner Good Smaritan Med Ctr, Tucson, AZ 85721 USA
[6] Arizona Alzheimers Dis Consortium, Phoenix, AZ USA
关键词
mitochondria; Alzheimer's disease; heteroplasmy; microarray; resequencing;
D O I
10.1016/j.mito.2006.07.002
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The role of mitochondrial dysfunction in the pathogenesis of Alzheimer's disease (AD) has been well documented. Though evidence for the role of mitochondria in AD seems incontrovertible, the impact of mitochondrial DNA (mtDNA) mutations in AD etiology remains controversial. Though mutations in mitochondrially encoded genes have repeatedly been implicated in the pathogenesis of AD, many of these studies have been, plagued by lack of replication as well as potential contamination of nuclear-encoded mitochondrial pseudogenes. To assess the role of mtDNA mutations in the pathogenesis of AD, while avoiding the pitfalls of nuclear-encoded mitochondrial pseudogenes encountered in previous investigations and showcasing the benefits of a novel resequencing technology, we sequenced the entire coding region (15,452 bp) of mtDNA from 19 extremely well-characterized AD patients and 18 age-matched, unaffected controls utilizing a new, reliable, high-throughput array-based resequencing technique, the Human MitoChip. High-throughput, array-based DNA resequencing of the entire mtDNA coding region from platelets of 37 subjects revealed the presence of 208 loci displaying a total of 917 sequence variants. There were no statistically significant differences in overall mutational burden between cases and controls, however, 265 independent sites of statistically significant change between cases and controls were identified. Changed sites were found in genes associated with complexes I (30.2%), III (3.0%), IV (33.2%), and V (9.1%) as well as tRNA (10.6%) and rRNA (14.0%). Despite their statistical significance, the subtle nature of the observed changes makes it difficult to determine whether they represent true functional variants involved in AD etiology or merely naturally occurring dissimilarity. Regardless, this study demonstrates the tremendous value of this novel mtDNA resequencing platform, which avoids the pitfalls of erroneously amplifying nuclear-encoded mtDNA pseudogenes, and our proposed analysis paradigm, which utilizes the availability of raw signal intensity values for each of the four potential alleles to facilitate quantitative estimates of mtDNA heteroplasmy. This information provides a potential new target for burgeoning diagnostics and therapeutics that could truly assist those suffering from this devastating disorder. (c) 2006 Elsevier B.V. and Mitochondria Research Society. All rights reserved.
引用
收藏
页码:194 / 210
页数:17
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