Genome-Wide Association Study of CSF Levels of 59 Alzheimer's Disease Candidate Proteins: Significant Associations with Proteins Involved in Amyloid Processing and Inflammation

English version České info

The use of quantitative endophenotypes from cerebrospinal fluid has led to the identification of several genetic variants that alter risk or rate of progression of Alzheimer's disease. Here we have analyzed the levels of 58 disease-related proteins in the cerebrospinal fluid for association with millions of variants across the human genome. We have identified significant, replicable associations with 5 analytes, Angiotensin-converting enzyme, Chemokine (C-C motif) ligand 2, Chemokine (C-C motif) ligand 4, Interleukin 6 receptor and Matrix metalloproteinase-3. Our results suggest that these variants play a regulatory role in the respective protein levels and are relevant to the inflammatory and amyloid processing pathways. Variants in associated with ACE and those associated with MMP3 levels also show association with risk for Alzheimer's disease in the expected directions. These associations are consistent in cerebrospinal fluid and plasma and in samples with only cognitively normal individuals suggesting that they are relevant in the regulation of these protein levels beyond the context of Alzheimer's disease.

Vyšlo v časopise: Genome-Wide Association Study of CSF Levels of 59 Alzheimer's Disease Candidate Proteins: Significant Associations with Proteins Involved in Amyloid Processing and Inflammation. PLoS Genet 10(10): e32767. doi:10.1371/journal.pgen.1004758
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004758

Souhrn

Zdroje

1. FaganAM, RoeCM, XiongC, MintunMA, MorrisJC, et al. (2007) Cerebrospinal fluid tau/beta-amyloid(42) ratio as a prediction of cognitive decline in nondemented older adults. Arch Neurol 64 : 343–349.

2. SniderBJ, FaganAM, RoeC, ShahAR, GrantEA, et al. (2009) Cerebrospinal fluid biomarkers and rate of cognitive decline in very mild dementia of the Alzheimer type. Arch Neurol 66 : 638–645.

3. HuangJT, WangL, PrabakaranS, WengenrothM, LockstoneHE, et al. (2008) Independent protein-profiling studies show a decrease in apolipoprotein A1 levels in schizophrenia CSF, brain and peripheral tissues. Mol Psychiatry 13 : 1118–1128.

4. BiblM, MollenhauerB, EsselmannH, LewczukP, KlafkiHW, et al. (2006) CSF amyloid-beta-peptides in Alzheimer's disease, dementia with Lewy bodies and Parkinson's disease dementia. Brain 129 : 1177–1187.

5. ShenL, ThompsonPM, PotkinSG, BertramL, FarrerLA, et al. (2013) Genetic analysis of quantitative phenotypes in AD and MCI: imaging, cognition and biomarkers. Brain Imaging Behav 8 : 183–207 doi:10.1007/s11682-013-9262-z

6. CruchagaC, EbbertMW, KauweJK (2014) Genetic Discoveries in AD Using CSF Amyloid and Tau. Current Genetic Medicine Reports 1–7.

7. PetersonD, MungerC, CrowleyJ, CorcoranC, CruchagaC, et al. (2013) Variants in PPP3R1 and MAPT are associated with more rapid functional decline in Alzheimer's disease: The Cache County Dementia Progression Study. Alzheimer's & Dementia 10 : 366–71 doi:10.1016/j.jalz.2013.02.010

8. PetersonD, MungerC, CrowleyJ, CorcoranC, CruchagaC, et al. (2013) Variants in PPP3R1 and MAPT are associated with more rapid functional decline in Alzheimer's disease: The Cache County Dementia Progression Study. Alzheimers Dement 10 : 366–71 doi:10.1016/j.jalz.2013.02.010

9. CruchagaC, KauweJS, HarariO, JinSC, CaiY, et al. (2013) GWAS of cerebrospinal fluid tau levels identifies risk variants for Alzheimer's disease. Neuron 78 : 256–268.

10. CruchagaC, KauweJS, NowotnyP, BalesK, PickeringEH, et al. (2012) Cerebrospinal fluid APOE levels: an endophenotype for genetic studies for Alzheimer's disease. Hum Mol Genet 21 : 4558–71.

11. KauweJS, CruchagaC, KarchCM, SadlerB, LeeM, et al. (2011) Fine mapping of genetic variants in BIN1, CLU, CR1 and PICALM for association with cerebrospinal fluid biomarkers for Alzheimer's disease. PLoS One 6: e15918.

12. KauweJS, CruchagaC, BertelsenS, MayoK, LatuW, et al. (2010) Validating predicted biological effects of Alzheimer's disease associated SNPs using CSF biomarker levels. J Alzheimers Dis 21 : 833–842.

13. CruchagaC, KauweJS, MayoK, SpiegelN, BertelsenS, et al. (2010) SNPs associated with cerebrospinal fluid phospho-tau levels influence rate of decline in Alzheimer's disease. PLoS Genet 6: e1001101 doi:10.1371/journal.pgen.1001101

14. KauweJS, WangJ, MayoK, MorrisJC, FaganAM, et al. (2009) Alzheimer's disease risk variants show association with cerebrospinal fluid amyloid beta. Neurogenetics 10 : 13–17.

15. KauweJS, CruchagaC, MayoK, FenoglioC, BertelsenS, et al. (2008) Variation in MAPT is associated with cerebrospinal fluid tau levels in the presence of amyloid-beta deposition. Proc Natl Acad Sci U S A 105 : 8050–8054.

16. KauweJS, JacquartS, ChakravertyS, WangJ, MayoK, et al. (2007) Extreme cerebrospinal fluid amyloid beta levels identify family with late-onset Alzheimer's disease presenilin 1 mutation. Ann Neurol 61 : 446–453.

17. BekrisLM, LutzF, YuCE (2012) Functional analysis of APOE locus genetic variation implicates regional enhancers in the regulation of both TOMM40 and APOE. J Hum Genet 57 : 18–25.

18. Elias-SonnenscheinLS, HelisalmiS, NatunenT, HallA, PaajanenT, et al. (2013) Genetic loci associated with Alzheimer's disease and cerebrospinal fluid biomarkers in a Finnish case-control cohort. PLoS One 8: e59676.

19. GiedraitisV, GlaserA, SarajarviT, BrundinR, GunnarssonMD, et al. (2010) CALHM1 P86L polymorphism does not alter amyloid-beta or tau in cerebrospinal fluid. Neurosci Lett 469 : 265–267.

20. HanMR, SchellenbergGD, WangLS, Alzheimer's Disease NeuroimagingI (2010) Genome-wide association reveals genetic effects on human Abeta42 and tau protein levels in cerebrospinal fluids: a case control study. BMC Neurol 10 : 90.

21. KimS, SwaminathanS, InlowM, RisacherSL, NhoK, et al. (2013) Influence of genetic variation on plasma protein levels in older adults using a multi-analyte panel. PLoS One 8: e70269.

22. KimS, SwaminathanS, ShenL, RisacherSL, NhoK, et al. (2011) Genome-wide association study of CSF biomarkers Abeta1–42, t-tau, and p-tau181p in the ADNI cohort. Neurology 76 : 69–79.

23. NhoK, CorneveauxJJ, KimS, LinH, RisacherSL, et al. (2013) Identification of functional variants from whole-exome sequencing, combined with neuroimaging genetics. Mol Psychiatry 18 : 739.

24. NhoK, CorneveauxJJ, KimS, LinH, RisacherSL, et al. (2013) Whole-exome sequencing and imaging genetics identify functional variants for rate of change in hippocampal volume in mild cognitive impairment. Mol Psychiatry 18 : 781–787.

25. RidgePG, KoopA, MaxwellTJ, BaileyMH, SwerdlowRH, et al. (2013) Mitochondrial haplotypes associated with biomarkers for Alzheimer's disease. PLoS One 8: e74158.

26. ThompsonPM, SteinJL, MedlandSE, HibarDP, VasquezAA, et al. (2014) The ENIGMA Consortium: large-scale collaborative analyses of neuroimaging and genetic data. Brain Imaging Behav

27. YoderKK, NhoK, RisacherSL, KimS, ShenL, et al. (2013) Influence of TSPO genotype on 11C-PBR28 standardized uptake values. J Nucl Med 54 : 1320–1322.

28. HoltzmanDM, MorrisJC, GoateAM (2011) Alzheimer's disease: the challenge of the second century. Sci Transl Med 3 : 77sr71.

29. ChungCM, WangRY, ChenJW, FannCS, LeuHB, et al. (2010) A genome-wide association study identifies new loci for ACE activity: potential implications for response to ACE inhibitor. Pharmacogenomics J 10 : 537–544.

30. LambertJC, Ibrahim-VerbaasCA, HaroldD, NajAC, SimsR, et al. (2013) Meta-analysis of 74,046 individuals identifies 11 new susceptibility loci for Alzheimer's disease. Nat Genet 45 : 1452–1458.

31. FerreiraMA, MathesonMC, DuffyDL, MarksGB, HuiJ, et al. (2011) Identification of IL6R and chromosome 11q13.5 as risk loci for asthma. Lancet 378 : 1006–1014.

32. DehghanA, DupuisJ, BarbalicM, BisJC, EiriksdottirG, et al. (2011) Meta-analysis of genome-wide association studies in >80 000 subjects identifies multiple loci for C-reactive protein levels. Circulation 123 : 731–738.

33. DaviesRW, WellsGA, StewartAF, ErdmannJ, ShahSH, et al. (2012) A genome-wide association study for coronary artery disease identifies a novel susceptibility locus in the major histocompatibility complex. Circ Cardiovasc Genet 5 : 217–225.

34. ChengYC, KaoWH, MitchellBD, O'ConnellJR, ShenH, et al. (2009) Genome-wide association scan identifies variants near Matrix Metalloproteinase (MMP) genes on chromosome 11q21-22 strongly associated with serum MMP-1 levels. Circ Cardiovasc Genet 2 : 329–337.

35. TolboomTC, PietermanE, van der LaanWH, ToesRE, HuidekoperAL, et al. (2002) Invasive properties of fibroblast-like synoviocytes: correlation with growth characteristics and expression of MMP-1, MMP-3, and MMP-10. Ann Rheum Dis 61 : 975–980.

36. DorrS, LechtenbohmerN, RauR, HerbornG, WagnerU, et al. (2004) Association of a specific haplotype across the genes MMP1 and MMP3 with radiographic joint destruction in rheumatoid arthritis. Arthritis research & therapy 6: R199–207.

37. O'ReillyPF, HoggartCJ, PomyenY, CalboliFCF, ElliottP, et al. (2012) MultiPhen: Joint Model of Multiple Phenotypes Can Increase Discovery in GWAS. PLoS ONE 7: e34861.

38. ObaR, IgarashiA, KamataM, NagataK, TakanoS, et al. (2005) The N-terminal active centre of human angiotensin-converting enzyme degrades Alzheimer amyloid beta-peptide. Eur J Neurosci 21 : 733–740.

39. HemmingML, SelkoeDJ (2005) Amyloid beta-protein is degraded by cellular angiotensin-converting enzyme (ACE) and elevated by an ACE inhibitor. J Biol Chem 280 : 37644–37650.

40. HuJ, IgarashiA, KamataM, NakagawaH (2001) Angiotensin-converting enzyme degrades Alzheimer amyloid beta-peptide (A beta); retards A beta aggregation, deposition, fibril formation; and inhibits cytotoxicity. J Biol Chem 276 : 47863–47868.

41. ZouK, YamaguchiH, AkatsuH, SakamotoT, KoM, et al. (2007) Angiotensin-converting enzyme converts amyloid beta-protein 1–42 (Abeta(1–42)) to Abeta(1–40), and its inhibition enhances brain Abeta deposition. J Neurosci 27 : 8628–8635.

42. HeM, OhruiT, MaruyamaM, TomitaN, NakayamaK, et al. (2006) ACE activity in CSF of patients with mild cognitive impairment and Alzheimer disease. Neurology 67 : 1309–1310.

43. NingM, YangY, ZhangZ, ChenZ, ZhaoT, et al. (2010) Amyloid-beta-Related Genes SORL1 and ACE are Genetically Associated With Risk for Late-onset Alzheimer Disease in the Chinese Population. Alzheimer disease and associated disorders

44. MinersJS, van HelmondZ, RaikerM, LoveS, KehoePG (2010) ACE variants and association with brain Abeta levels in Alzheimer's disease. Am J Transl Res 3 : 73–80.

45. HelbecqueN, CodronV, CottelD, AmouyelP (2009) An age effect on the association of common variants of ACE with Alzheimer's disease. Neurosci Lett 461 : 181–184.

46. MengY, BaldwinCT, BowirratA, WaraskaK, InzelbergR, et al. (2006) Association of polymorphisms in the Angiotensin-converting enzyme gene with Alzheimer disease in an Israeli Arab community. Am J Hum Genet 78 : 871–877.

47. KehoePG, KatzovH, AndreasenN, GatzM, WilcockGK, et al. (2004) Common variants of ACE contribute to variable age-at-onset of Alzheimer's disease. Human genetics 114 : 478–483.

48. BelbinO, BrownK, ShiH, MedwayC, AbrahamR, et al. (2011) A multi-center study of ACE and the risk of late-onset Alzheimer's disease. Journal of Alzheimer's disease: JAD 24 : 587–597.

49. BruandetA, RichardF, TzourioC, BerrC, DartiguesJF, et al. (2008) Haplotypes across ACE and the risk of Alzheimer's disease: the three-city study. Journal of Alzheimer's disease: JAD 13 : 333–339.

50. HarariO, CruchagaC, KauweJS, AinscoughBJ, BalesK, et al. (2014) Ptau-Aβ42 ratio as a continuous trait for biomarker discovery for early stage Alzheimer's disease in multiplex immunoassay panels of Cerebrospinal fluid. Biological Psychiatry 75 : 723–731.

51. YoshiyamaY, SatoH, SeikiM, ShinagawaA, TakahashiM, et al. (1998) Expression of the membrane-type 3 matrix metalloproteinase (MT3-MMP) in human brain tissues. Acta Neuropathol 96 : 347–350.

52. YoshiyamaY, AsahinaM, HattoriT (2000) Selective distribution of matrix metalloproteinase-3 (MMP-3) in Alzheimer's disease brain. Acta Neuropathol 99 : 91–95.

53. DebS, GottschallPE (1996) Increased production of matrix metalloproteinases in enriched astrocyte and mixed hippocampal cultures treated with beta-amyloid peptides. J Neurochem 66 : 1641–1647.

54. WhiteAR, DuT, LaughtonKM, VolitakisI, SharplesRA, et al. (2006) Degradation of the Alzheimer disease amyloid beta-peptide by metal-dependent up-regulation of metalloprotease activity. J Biol Chem 281 : 17670–17680.

55. YinKJ, CirritoJR, YanP, HuX, XiaoQ, et al. (2006) Matrix metalloproteinases expressed by astrocytes mediate extracellular amyloid-beta peptide catabolism. J Neurosci 26 : 10939–10948.

56. YanP, HuX, SongH, YinK, BatemanRJ, et al. (2006) Matrix metalloproteinase-9 degrades amyloid-beta fibrils in vitro and compact plaques in situ. J Biol Chem 281 : 24566–24574.

57. BackstromJR, LimGP, CullenMJ, TokesZA (1996) Matrix metalloproteinase-9 (MMP-9) is synthesized in neurons of the human hippocampus and is capable of degrading the amyloid-beta peptide (1–40). J Neurosci 16 : 7910–7919.

58. ReitzC, van RooijFJ, de MaatMP, den HeijerT, HofmanA, et al. (2008) Matrix metalloproteinase 3 haplotypes and dementia and Alzheimer's disease. The Rotterdam Study. Neurobiology of aging 29 : 874–881.

59. FlexA, GaetaniE, ProiaAS, PecoriniG, StrafaceG, et al. (2006) Analysis of functional polymorphisms of metalloproteinase genes in persons with vascular dementia and Alzheimer's disease. J Gerontol A Biol Sci Med Sci 61 : 1065–1069.

60. SaarelaMS, LehtimakiT, RinneJO, HervonenA, JylhaM, et al. (2004) Interaction between matrix metalloproteinase 3 and the epsilon4 allele of apolipoprotein E increases the risk of Alzheimer's disease in Finns. Neurosci Lett 367 : 336–339.

61. ConductierG, BlondeauN, GuyonA, NahonJL, RovereC (2010) The role of monocyte chemoattractant protein MCP1/CCL2 in neuroinflammatory diseases. J Neuroimmunol 224 : 93–100.

62. NaertG, RivestS (2011) CC chemokine receptor 2 deficiency aggravates cognitive impairments and amyloid pathology in a transgenic mouse model of Alzheimer's disease. J Neurosci 31 : 6208–6220.

63. KiyotaT, YamamotoM, XiongH, LambertMP, KleinWL, et al. (2009) CCL2 accelerates microglia-mediated Abeta oligomer formation and progression of neurocognitive dysfunction. PLoS One 4: e6197.

64. SeveriniC, PasseriPP, CiottiM, FlorenzanoF, PossentiR, et al. (2014) Bindarit, inhibitor of CCL2 synthesis, protects neurons against amyloid-beta-induced toxicity. J Alzheimers Dis 38 : 281–293.

65. Wyss-CorayT, LoikeJD, BrionneTC, LuE, AnankovR, et al. (2003) Adult mouse astrocytes degrade amyloid-beta in vitro and in situ. Nat Med 9 : 453–457.

66. GalimbertiD, FenoglioC, LovatiC, VenturelliE, GuidiI, et al. (2006) Serum MCP-1 levels are increased in mild cognitive impairment and mild Alzheimer's disease. Neurobiol Aging 27 : 1763–1768.

67. WestinK, BuchhaveP, NielsenH, MinthonL, JanciauskieneS, et al. (2012) CCL2 is associated with a faster rate of cognitive decline during early stages of Alzheimer's disease. PLoS ONE 7: e30525.

68. BoniniJA, MartinSK, DralyukF, RoeMW, PhilipsonLH, et al. (1997) Cloning, expression, and chromosomal mapping of a novel human CC-chemokine receptor (CCR10) that displays high-affinity binding for MCP-1 and MCP-3. DNA Cell Biol 16 : 1249–1256.

69. CrosslinDR, McDavidA, WestonN, ZhengX, HartE, et al. (2013) Genetic variation associated with circulating monocyte count in the eMERGE Network. Hum Mol Genet 22 : 2119–2127.

70. WilliamsDW, CalderonTM, LopezL, Carvallo-TorresL, GaskillPJ, et al. (2013) Mechanisms of HIV Entry into the CNS: Increased Sensitivity of HIV Infected CD14(+)CD16(+) Monocytes to CCL2 and Key Roles of CCR2, JAM-A, and ALCAM in Diapedesis. PLoS One 8: e69270.

71. VestergaardC, JustH, Baumgartner NielsenJ, Thestrup-PedersenK, DeleuranM (2004) Expression of CCR2 on monocytes and macrophages in chronically inflamed skin in atopic dermatitis and psoriasis. Acta dermato-venereologica 84 : 353–358.

72. McInnesIB, SchettG (2007) Cytokines in the pathogenesis of rheumatoid arthritis. Nature reviews Immunology 7 : 429–442.

73. BarlicJ, MurphyPM (2007) Chemokine regulation of atherosclerosis. Journal of leukocyte biology 82 : 226–236.

74. XiaMQ, QinSX, WuLJ, MackayCR, HymanBT (1998) Immunohistochemical study of the beta-chemokine receptors CCR3 and CCR5 and their ligands in normal and Alzheimer's disease brains. The American journal of pathology 153 : 31–37.

75. SmitsHA, RijsmusA, van LoonJH, WatJW, VerhoefJ, et al. (2002) Amyloid-beta-induced chemokine production in primary human macrophages and astrocytes. Journal of neuroimmunology 127 : 160–168.

76. AkiyamaH, BargerS, BarnumS, BradtB, BauerJ, et al. (2000) Inflammation and Alzheimer's disease. Neurobiol Aging 21 : 383–421.

77. LiangosO, AddabboF, TighiouartH, GoligorskyM, JaberBL (2010) Exploration of disease mechanism in acute kidney injury using a multiplex bead array assay: a nested case-control pilot study. Biomarkers: biochemical indicators of exposure, response, and susceptibility to chemicals 15 : 436–445.

78. ModiWS, LautenbergerJ, AnP, ScottK, GoedertJJ, et al. (2006) Genetic variation in the CCL18-CCL3-CCL4 chemokine gene cluster influences HIV Type 1 transmission and AIDS disease progression. American journal of human genetics 79 : 120–128.

79. CampbellIL, ErtaM, LimSL, FraustoR, MayU, et al. (2014) Trans-signaling is a dominant mechanism for the pathogenic actions of interleukin-6 in the brain. J Neurosci 34 : 2503–2513.

80. FerreiraRC, FreitagDF, CutlerAJ, HowsonJM, RainbowDB, et al. (2013) Functional IL6R 358Ala allele impairs classical IL-6 receptor signaling and influences risk of diverse inflammatory diseases. PLoS Genet 9: e1003444.

81. QiL, RifaiN, HuFB (2007) Interleukin-6 receptor gene variations, plasma interleukin-6 levels, and type 2 diabetes in U.S. Women. Diabetes 56 : 3075–3081.

82. ChalarisA, GewieseJ, PaligaK, FleigL, SchneedeA, et al. (2010) ADAM17-mediated shedding of the IL6R induces cleavage of the membrane stub by gamma-secretase. Biochim Biophys Acta 1803 : 234–245.

83. WangM, SongH, JiaJ (2010) Interleukin-6 receptor gene polymorphisms were associated with sporadic Alzheimer's disease in Chinese Han. Brain research 1327 : 1–5.

84. HampelH, SunderlandT, KotterHU, SchneiderC, TeipelSJ, et al. (1998) Decreased soluble interleukin-6 receptor in cerebrospinal fluid of patients with Alzheimer's disease. Brain research 780 : 356–359.

85. Del BoR, AngerettiN, LuccaE, De SimoniMG, ForloniG (1995) Reciprocal control of inflammatory cytokines, IL-1 and IL-6, and beta-amyloid production in cultures. Neuroscience letters 188 : 70–74.

86. VandenabeeleP, FiersW (1991) Is amyloidogenesis during Alzheimer's disease due to an IL-1-/IL-6-mediated ‘acute phase response’ in the brain? Immunology today 12 : 217–219.

87. RingheimGE, SzczepanikAM, PetkoW, BurgherKL, ZhuSZ, et al. (1998) Enhancement of beta-amyloid precursor protein transcription and expression by the soluble interleukin-6 receptor/interleukin-6 complex. Brain research Molecular brain research 55 : 35–44.

88. Sabater-LlealM, HuangJ, ChasmanD, NaitzaS, DehghanA, et al. (2013) Multiethnic meta-analysis of genome-wide association studies in >100 000 subjects identifies 23 fibrinogen-associated Loci but no strong evidence of a causal association between circulating fibrinogen and cardiovascular disease. Circulation 128 : 1310–1324.

89. MelzerD, PerryJR, HernandezD, CorsiAM, StevensK, et al. (2008) A genome-wide association study identifies protein quantitative trait loci (pQTLs). PLoS Genet 4: e1000072.

90. WilkJB, WalterRE, LaramieJM, GottliebDJ, O'ConnorGT (2007) Framingham Heart Study genome-wide association: results for pulmonary function measures. BMC Med Genet 8 Suppl 1: S8.

91. ChauhanD, UchiyamaH, AkbaraliY, UrashimaM, YamamotoK, et al. (1996) Multiple myeloma cell adhesion-induced interleukin-6 expression in bone marrow stromal cells involves activation of NF-kappa B. Blood 87 : 1104–1112.

92. IshiharaK, HiranoT (2002) IL-6 in autoimmune disease and chronic inflammatory proliferative disease. Cytokine & growth factor reviews 13 : 357–368.

93. TanD, WuX, HouM, LeeSO, LouW, et al. (2005) Interleukin-6 polymorphism is associated with more aggressive prostate cancer. The Journal of urology 174 : 753–756.

94. TrojanowskiJQ, VandeersticheleH, KoreckaM, ClarkCM, AisenPS, et al. (2010) Update on the biomarker core of the Alzheimer's Disease Neuroimaging Initiative subjects. Alzheimers Dement 6 : 230–238.

95. FaganAM, MintunMA, MachRH, LeeSY, DenceCS, et al. (2006) Inverse relation between in vivo amyloid imaging load and cerebrospinal fluid Abeta42 in humans. Ann Neurol 59 : 512–519.

96. PriceAL, PattersonNJ, PlengeRM, WeinblattME, ShadickNA, et al. (2006) Principal components analysis corrects for stratification in genome-wide association studies. Nat Genet 38 : 904–909.

97. BrowningSR (2008) Missing data imputation and haplotype phase inference for genome-wide association studies. Hum Genet 124 : 439–450.

98. PurcellS, NealeB, Todd-BrownK, ThomasL, FerreiraMA, et al. (2007) PLINK: a tool set for whole-genome association and population-based linkage analyses. Am J Hum Genet 81 : 559–575.

99. AbecasisG, WillerC (2007) Metal—Meta Analysis Helper. Center for Statistical Genetics

100. AulchenkoYS, RipkeS, IsaacsA, van DuijnCM (2007) GenABEL: an R library for genome-wide association analysis. Bioinformatics 23 : 1294–1296.

101. JagustWJ, LandauSM, ShawLM, TrojanowskiJQ, KoeppeRA, et al. (2009) Relationships between biomarkers in aging and dementia. Neurology 73 : 1193–1199.

102. WangK, LiM, HakonarsonH (2010) ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res 38: e164.

103. BoyleAP, HongEL, HariharanM, ChengY, SchaubMA, et al. (2012) Annotation of functional variation in personal genomes using RegulomeDB. Genome Res 22 : 1790–1797.

104. NgPC, HenikoffS (2003) SIFT: Predicting amino acid changes that affect protein function. Nucleic Acids Res 31 : 3812–3814.

105. AdzhubeiIA, SchmidtS, PeshkinL, RamenskyVE, GerasimovaA, et al. (2010) A method and server for predicting damaging missense mutations. Nat Methods 7 : 248–249.