Meta-analysis of 49 549 individuals imputed with the 1000 Genomes Project reveals an exonic damaging variant in ANGPTL4 determining fasting TG levels

SHORT REPORT Me a-analysis o 49 549 indi iduals impu ed wi h he 1000 Genomes P ojec e eals an exonic damaging a ian in ANGPTL4 de e mining as ing TG le els Elisabe h M an Leeuwen, 1 Aniko Sabo, 2 Joshua C Bis, 3 Jenni e E Hu man, 4,5 Ani Manichaikul, 6 Albe V Smi h, 7,8 Ma y F Fei osa, 9 Se kalem Demissie, 10 Pe e K Joshi, 11 Qing Duan, 12 Jona han Ma en, 4 Jan B an Klinken, 13 Ida Su akka, 14 Ilja M Nol e, 15 Weihua Zhang, 16,17 Hamdi Mba ek, 18 Rui ang Li-Gao, 19 S ella T ompe , 20,21 Niek Ve weij, 22 E angelos E angelou, 16,23 Leo-Pekka Lyy ikäinen, 24,25 Bamidele O Tayo, 26 Jo is Deelen, 27 Pe e J an de Mos , 15 Sande W an de Laan, 28 Dan E A king, 29 Alanna Mo ison, 30 Abbas Dehghan, 1 Osca H F anco, 1 Albe Ho man, 1 Fe nando Ri adenei a, 31 E ic J Sijb ands, 31 And e G Ui e linden, 1,31 Josy C Mychaleckyj, 6 A chie Campbell, 32 Lynne J Hocking, 33 Sandosh Padmanabhan, 34 Jenni e A B ody, 3 Kenne h M Rice, 35 Cha les C Whi e, 36 Tama a Ha is, 37 Aa on Isaacs, 1 Ha y Campbell, 11 Leslie A Lange, 12 Igo Rudan, 38 I ana Kolcic, 39 Pau Na a o, 4 Ta ijana Zemunik, 39 Veikko Salomaa, 40 The Li eLines Coho S udy Angad S Koone , 41 Jaspal S Koone , 17,41,42 Benjamin Lehne, 16 William R Sco , 16,17 Sian-Tsung Tan, 41 Eco J de Geus, 18 Yu i Milaneschi, 43 B enda W J H Penninx, 43 Gonneke Willemsen, 18 Renée de Mu se , 19 Ian Fo d, 44 Ron T Ganse oo , 45 Ma celo P Segu a-Lepe, 16 Olli T Rai aka i, 46,47 Jo ma S Viika i, 48,49 Kjell Nikus, 50,51 Te ence Fo es e , 52 Colin A McKenzie, 52 An on J M de C aen, 21 Hes e M de Ruij e , 28 CHARGE Lipids Wo king G oup Ge a d Pas e kamp, 28,53 Ha old Sniede , 15 Albe ine J Oldehinkel, 54 P Eline Slagboom, 27 Richa d S Coope , 26 Mika Kähönen, 55,56 Te ho Leh imäki, 24,25 Paul Ellio , 57 Pim an de Ha s , 22,58 J Wou e Jukema, 20 Dennis O Mook-Kanamo i, 19,59,60 Do e I Boomsma, 18 John C Chambe s, 16,17,42 Mo is Swe z, 58,61 Samuli Ripa i, 14,62,63 Ko Willems an Dijk, 13,64 Ve onique Vi a , 4 Oz en Polasek, 39 Ca oline Haywa d, 4 James G Wilson, 65 James F Wilson, 4,11 Vilmundu Gudnason, 7,8 S ephen S Rich, 6 B uce M Psa y, 3,66,67,68 Ing id B Bo ecki, 9 E ic Boe winkle, 2,30 Je ome I Ro e , 69,70,71 L Ad ienne Cupples, 5,9 Co nelia M an Duijn 1 ▸Addi ional ma e ial is published online only. To iew his file please isi he jou nal online (h p://dx.doi.o g/10. 1136/jmedgene -2015- 103439) Fo numbe ed a filia ions see end o a icle. Co espondence o P o esso Co nelia M an Duijn, Gene ic Epidemiology Uni , Depa men o Epidemiology, E asmus Medical Cen e , Pos bus 2040, Ro e dam 3000 CA, The Ne he lands; c. anduijn@e asmusmc.nl Recei ed 4 Augus 2015 Re ised 19 No embe 2015 Accep ed 23 No embe 2015 Published Online Fi s 1 Ap il 2016 To ci e: an Leeuwen EM, Sabo A, Bis JC, e al.J Med Gene 2016;53:441–449. ABSTRACT Backg ound So a , mo e han 170 loci ha e been associa ed wi h ci cula ing lipid le els h ough genome- wide associa ion s udies (GWAS). These associa ions a e la gely d i en by common a ian s, hei unc ion is o en no known, and many a e likely o be ma ke s o he causal a ian s. In his s udy we aimed o iden i y mo e new a e and low- equency unc ional a ian s associa ed wi h ci cula ing lipid le els. Me hods We used he 1000 Genomes P ojec as a e e ence panel o he impu a ions o GWAS da a om ∼60 000 indi iduals in he disco e y s age and ∼90 000 samples in he eplica ion s age. Resul s Ou s udy esul ed in he iden ifica ion o fi e new associa ions wi h ci cula ing lipid le els a ou loci. All ou loci a e wi hin genes ha can be linked biologically o lipid me abolism. One o he a ian s, s116843064, is a damaging missense a ian wi hin he ANGPTL4 gene. Conclusions This s udy illus a es ha GWAS wi h high-scale impu a ion may s ill help us un a el he biological mechanism behind ci cula ing lipid le els. INTRODUCTION Genome-wide associa ion s udies (GWAS) o ci cu- la ing lipid le els (high-densi y lipop o ein choles- e ol (HDL-C), low-densi y lipop o ein choles e ol (LDL-C), o al choles e ol (TC) and iglyce ides (TG)) ha e iden ified o e 170 loci. 1–3 These Open Access Scan o access mo e ee con en an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 441 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om s udies ha e been based on impu a ions o he HapMap e e - ence panel 2 o p ima y e sions o he 1000 Genomes P ojec (1kG) 1 o geno yping on he Illumina Exome Chip. 3 None has used impu a ions wi h he Phase 1 in eg a ed elease 3 o he 1kG which allows he impu a ion o a e and low- equency unc ional a ian s and s uc u al a ia ions wi h mo e p eci- sion. E idence o a e and low- equency unc ional a ian s associa ed wi h ci cula ing lipid le els comes om ecen s udies in which exome sequencing o he NPC1L1 gene iden ified a e a ian s associa ed wi h educed LDL-C le els and educed isk o co ona y hea disease. 4 Mo eo e , exome sequencing o LDLR and APOA5 iden ified a e a ian s associa ed wi h an inc eased LDL-C and inc eased TG le els 5 and exome sequen- cing o APOC3 iden ified a e a ian s associa ed wi h educed TG le els and educed isk o co ona y hea disease. 6 Ou goal in his s udy was o iden i y a e and low- equency unc ional a ian s associa ed wi h ci cula ing lipid le els in a la ge sample size compa ed wi h he exome sequencing o can- dida e gene app oach. To his end, we impu ed geno ypes o s udy samples pa icipa ing in he coho s o he Coho s o Hea and Aging Resea ch in Genomic Epidemiology (CHARGE) conso ium using he Phase 1 in eg a ed elease V. 3 o he 1kG and conduc ed a me a-analysis o abou app oxi- ma ely 60 000 indi iduals, ollowed by a eplica ion in an inde- penden se o 90 000 indi iduals. METHODS Please see online supplemen a y me hods o comple e desc ip- ions o he me hods. In summa y, o he disco e y s age o his p ojec , we used he da a om 20 coho s o he CHARGE con- so ium (see online supplemen a y me hods). All coho s we e impu ed wi h e e ence o he 1kG e e ence panel ( e sion Phase 1 in eg a ed elease V.3). The o al numbe o indi iduals in he disco e y s age was 59 409 o HDL-C, 48 780 o LDL-C, 60 024 o TC and 49 549 o TG. Online supplemen- a y ables S1 and S2 con ain he baseline cha ac e is ics pe coho and mo e de ails abou SNP geno yping and geno ype impu a ions.Wi hin each coho , each a ian was es ed o associa ion wi h each o he lipid ai s, assuming an addi i e gene ic model. The associa ion esul s o all coho s o all a - ian s we e combined using in e se a iance weigh ing. We used he ollowing fil e s o he a ian s: 0.3<R 2 (measu emen o he impu a ion quali y) ≤1.0 and expec ed mino allele coun (expMAC=2×MAF (mino allele equency)×R 2 ×sample size) >10 p io o me a-analysis. A e me a-analysis o all a ailable a ian s, we excluded he a ian s ha we e no p esen in a leas ou coho s, o p e en alse posi i e findings. In o de o selec only a ian s ha we e independen ly associa ed wi h each o he lipid ai s, we used he genome-wide complex ai analysis (GCTA) 7 ool, V.1.13. To iden i y no el loci we selec ed om he lis o a ian s iden ified by GCTA, hose a ian s loca ed mo e han 0.5 Mb away om p e iously iden ified loci o he co esponding ai 23 and which we e significan (p alue<5×10 −8 ) in he ini ial disco e y s age. To p e en he iden ifica ion o alse posi i e loci, we added a second eplica- ion s age wi hin 23 independen coho s. The expe imen -wide significance h eshold equi ed o keep ype I e o a e wi hin he eplica ion s age a 5% is 2.63×10 −3 (Bon e oni co ec ion based on 19 a ian s). We also me a-analysed he indi iduals o he disco e y and eplica ion s age oge he and pe e hnici y using a fixed-e ec app oach. We also epea ed his analysis wi h genome-wide associa ion me a analysis (GWAMA) (V.2.0.5) using a andom e ec app oach as he indi iduals in disco e y and eplica ion s ages come om mul iple e hnici ies. RESULTS The associa ion o all a ian s wi h HDL-C, LDL-C, TC and TG was es ed in all disco e y coho s (see online supplemen- a y figu es S1 and S2). The associa ion esul s o all disco e y coho s o all a ian s we e combined in a fixed-e ec me a-analysis using METAL (see online supplemen a y figu es S3 and S4). We significan ly eplica ed 88.1% o he loci desc ibed by Teslo ich e al 2 despi e a sample size o abou 80% (see online supplemen a y figu e S5 and supplemen a y able S3). We also significan ly eplica ed 43.4% o he loci desc ibed by he Global Lipids Gene ics Conso ium (GLGC) 3 despi e a sample size o abou 30% (see online supplemen a y figu e S6 and supplemen a y able S4). A condi ional and join analysis using GCTA iden ified 185 independen a ian s o HDL-C, 174 o LDL-C, 214 o TC and 119 o TG. Nex , we excluded all a ian s ha we e no genome-wide significan (p alue<5×10 −8 ) in he ini ial disco - e y s age, which esul ed in 56 a ian s o HDL-C, 50 o LDL-C, 66 o TC and 37 o TG. And we excluded all a ian s which a e wi hin 0.5 Mb o a loci p e iously published by Teslo ich e al 2 o GLGC, 3 which esul ed in h ee a ian s o HDL-C, h ee o LDL-C, se en o TC and six o TG. These a ian s a e loca ed a 17 di e en loci and include one dele ion (figu e 1 and able 1). These 19 a ian s we e selec ed o eplica ion. The o al numbe o indi iduals in he eplica ion s age was 84 598, 72 486, 83 739 and 73 519 o HDL-C, LDL-C, TC and TG, espec i ely (see online supplemen a y ables S1 and S2 o baseline cha ac e is ics and in o ma ion abou SNP geno yping and impu a ion de ails).The sample size in he eplica ion s age was la ge han he ini ial disco e y sample o 17 ou o he 19 a ian s. The equencies o he a ian s we e simila be ween he disco e y and eplica ion coho s. The di ec ions o e ec we e he same in he disco e y and eplica ion coho s o 16 ou o he 19 a ian s (see online supplemen a y figu e S7). We used a Bon e oni co ec ed h eshold o significance (p alue<2.63×10 −3 ). Fi e ou o he 19 a ian s we e significan ly eplica ed ( able 1): s6457374 (TC), s186696265 (LDL-C and TC), s77697917 (HDL-C) and s116843064 (TG). The equency o hese a ian s anged be ween 0.012 and 0.249 wi hin he disco e y sample. Online supplemen a y able S5 shows he he e ogenei y o he 19 a ian s a e he me a-analysis o all disco e y coho s and o all eplica ion coho s. We also me a-analysed all a ian s in he indi iduals o he disco e y coho s and eplica ion coho s combined ( able 1 and see online supplemen a y ables S5 and S6) and pe e hnici y (see online supplemen a y able S6) using a fixed-e ec me a-analysis app oach. We ound ha he fi e significan ly eplica ed a ian s we iden ified in his s udy a e only significan wi hin he Eu opean samples, he eby no icing ha he e a e much mo e Eu opean samples in his s udy, compa ed wi h he A ican and Asian samples. When using a andom-e ec me a-analysis o accoun o he mul iple e hnici ies in ou sample (see online sup- plemen a y able S7), we ound ha o he fi e eplica ed a ian s, one a ained genome-wide significance (p alue<5×10 −8 ) and he o he ou nominal significance (p alue<0.05). DISCUSSION We conduc ed a GWAS ha included GWAS da a impu ed o he 1kG o iden i y a e and low- equency, po en ially unc- ional, a ian s associa ed wi h ci cula ing lipid le els. To his end, we impu ed geno ypes in app oxima ely 60 000 indi iduals om 20 coho s in he CHARGE conso ium wi h he 1kG 442 an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om e e ence panel. The me a-analysis, ollowed by GCTA analysis e ealed 19 associa ions wi h MAF anging om 0.01 o 0.48. O he 19 associa ions, we we e able o eplica e fi e in an inde- penden sample o app oxima ely 90 000 indi iduals. One o he fi e associa ions we iden ified is be ween TG and s116843064, an exonic a ian in he ANGPTL4 gene on ch omosome 19 (figu e 2C). This missense a ian changes he amino acid glu amic acid in o lysine (Glu40Lys) and is p edic ed o be damaging o he s uc u e and unc ion o he p o ein by Polyphen2, 8 Mu a ionTas e 9 and likelihood a io es (LRT). 10 ANGPTL4 is significan ly associa ed wi h he Kyo o Encyclopedia o Genes and Genomes (KEGG) e m a y acid me abolism, he GO p ocess lipid s o age and he gene on ology (GO) cellula componen lipid pa icle (p alue o 1.10×10 −6 , 1.31×10 −10 and 2.87×10 −18 , espec i ely, genene wo k.nl). ANGPTL4 has been associa ed wi h HDL-C be o e using he GWAS app oach 2 and wi h TG be o e using an exome sequencing app oach 11 and mo e ecen ly using he GWAS app oach. 1 We he e o e do no claim his finding as no el, hough his is he smalles s udy in which his a ian was genome-wide significan ly associa ed wi h TG and eplica ed in an independen sample. The second new finding we iden ified is he associa ion be ween TC and s6457374, an in e genic a ian loca ed on ch omosome 6 be ween he genes HLA-C and HLA-B (figu e 2A). Bo h genes a e associa ed wi h he KEGG e m ATP binding casse e (ABC) anspo e s (p alue o 4.29×10 −5 and 3.84×10 −5 o HLA-C and HLA-B, espec i ely, genene wo k. nl) which is in line wi h, among o he s, a p e iously published associa ion be ween TC and an exonic a ian in he ABCA6 gene which is also an ABC anspo e . 12 ABC anspo e s anspo a wide a ie y o subs a es ac oss ex acellula and in acellula memb anes, including lipids. 13 The hi d finding o his s udy is he associa ion be ween HDL-C and s77697917, an in e genic a ian on ch omosome 17 be ween he genes SOST and DUSP3 (figu e 2B). DUSP3 is associa ed wi h he egula ion and unc ion o ca bohyd a e- esponsi e elemen -binding p o ein (ChREBP) in he li e (p alue=3.03×10 −5 , genene wo k.nl). ChREBP med- ia es he ac i a ion o se e al egula o y enzymes in ol ed in lipogenesis. 14–18 This a ian is in high linkage disequilib ium (D0=0.936) in he 1 kG wi h s72836561, an exonic a ian in he gene CD300LG (MAF=0.027, β=−2.437, se β =0.381, p alue=1.51×10 −10 in he disco e y s age). This missense a ian changes he amino acid a ginine in o cys eine (A g82Cys) and is p edic ed o be damaging o he s uc u e and unc ion o he p o ein by Polyphen2, 8 Mu a ionTas e 9 and LRT. 10 This amino acid polymo phism has been associa ed wi h HDL-C in exome-wide associa ion s udies 19 and TG in GWAS 1 be o e. The ou h a ian we iden ified is s186696265, which is loca ed on ch omosome 6 and associa ed wi h LDL-C and TC (figu e 2D, E). This in e genic a ian is be ween he LPA (Lipop o ein, Lp(A)) gene and he PLG (Plasminogen) gene. The LPA gene has been associa ed be o e wi h LDL-C and TC be o e. 2 The epo ed lead SNP was s1564348, which in he newe human genome e sions is anno a ed o he SLC22A1 (Solu e Ca ie Family 22 (O ganic Ca ion T anspo e ), Membe 1) gene ins ead o he LPA gene. This explains why we again iden ified a locus nea he LPA gene, which has been iden- ified by o he s as well. 1 Fou een ou o he 19 a ian s we e no eplica ed despi e simila sample sizes and simila equencies wi hin he eplica- ion s age as compa ed wi h he disco e y s age. O hose 14 a ian s, 11 exhibi ed e ec sizes in he same di ec ion in bo h s ages. A possible explana ion migh be ha he eplica ion sample size is much la ge compa ed wi h ha o he disco e y sample size. Two a ian s migh ha e lacked significan eplica- ion due o small sample size, s60839105 and s151198427. Figu e 1 Manha an plo s o HDL-C (A), LDL-C (B), TC (C) and TG (D) a e he me a-analysis o all disco e y coho s. Va ian s ha we e p esen in a leas ou coho s and ha a e no wi hin 0.5 Mb o a p e iously published loci 23 we e included. The black line indica es he genome-wide significan line (5×10 −8 ), he black and ed do s he a ian s iden ified by GCTA which a e no genome-wide significan and which a e genome-wide significan , espec i ely. HDL-C, high-densi y lipop o ein choles e ol; LDL-C, low-densi y lipop o ein choles e ol; TC, o al choles e ol; TG, iglyce ides. an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 443 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om Table 1 The esul s o he 19 a ian s a e he me a-analysis o all disco e y coho s, all eplica ion coho s and all coho s combined Disco e y coho s Replica ion coho s All coho s combined T ai Ch :Posi ion s iden i ie nea es gene A1/A2 F eq N βSE β p Value F eq N βSE β p Value F eq βSE β p Value HDL-C 3:72 067 255 s75909755 PROL2-EIF4E3 T/C 0.03 62 607 1.593 0.275 7.27E-09 0.03 86 252 −0.019 0.031 5.45E-01 0.03 0.002 0.031 9.57E-01 TC 6:31 272 261 s6457374 HLA-B T/C 0.75 46 839 2.339 0.339 5.32E-12 0.81 74 417 0.057 0.016 4.23E-04 0.81 0.062 0.016 1.18E-04 LDL-C 6:31 325 323 s9266229 HLA-B C/G 0.53 37 981 −2.201 0.344 1.62E-10 0.41 61 582 −0.025 0.014 7.37E-02 0.41 −0.029 0.014 4.04E-02 TG 6:36 648 275 –CDKN1A CAG/C 0.45 53 425 −0.019 0.003 7.63E-09 0.49 59 018 −0.003 0.004 5.20E-01 0.46 −0.013 0.003 5.93E-07 TG 6:13 983 949 8 s608736 –C/G 0.48 53 425 −0.019 0.003 5.67E-09 0.49 73 512 −0.008 0.003 2.67E-02 0.48 −0.013 0.002 9.10E-09 TG 6:16 085 176 6 s376563 SLC22A3 T/C 0.46 47 036 −0.02 0.003 3.37E-09 0.46 73 512 −0.001 0.003 8.22E-01 0.46 −0.010 0.002 1.36E-05 LDL-C 6:16 111 170 0 s186696265 LPA-PLG T/C 0.01 49 221 11.247 1.241 1.31E-19 0.01 59 497 0.263 0.076 5.42E-04 0.01 0.304 0.076 6.17E-05 TC 6:16 111 170 0 s186696265 LPA-PLG T/C 0.01 59 859 10.004 1.162 7.20E-18 0.01 75 821 0.238 0.075 1.46E-03 0.01 0.278 0.075 1.93E-04 HDL-C 7:80 492 357 s60839105 SEMA3C T/C 0.07 7882 3.355 0.571 4.26E-09 0.08 4971 1.067 1.228 3.85E-01 0.07 2.948 0.518 1.25E-08 TC 8:68 351 787 s151198427 CPA6 A/G 0.11 17 361 6.552 1.147 1.12E-08 0.13 1419 −2.858 2.396 2.33E-01 0.11 4.797 1.035 3.56E-06 LDL-C 9:78 728 065 s146369471 PCSK5 T/C 0.99 43 398 8.529 1.449 3.99E-09 0.99 51 367 0.068 0.103 5.11E-01 0.99 0.110 0.103 2.84E-01 TC 9:78 728 065 s146369471 PCSK5 T/C 0.99 53 787 7.978 1.413 1.64E-08 0.99 70 241 0.015 0.103 8.84E-01 0.99 0.057 0.103 5.79E-01 TC 12:51 207 704 s829112 ATF1 A/G 0.68 56 924 1.448 0.258 2.02E-08 0.73 87 659 0.009 0.012 4.63E-01 0.73 0.012 0.012 3.18E-01 TG 13:11 454 402 4 s7140110 GAS6 T/C 0.71 48 221 −0.021 0.004 3.65E-08 0.72 60 437 −0.006 0.005 2.68E-01 0.72 −0.015 0.003 5.13E-07 TG 15:43 726 625 s150844304 TP53BP1 A/C 0.97 52 720 −0.083 0.01 2.52E-17 0.95 63 884 −0.026 0.015 8.85E-02 0.96 −0.066 0.008 9.52E-16 TC 17:18 046 290 s8065026 MYO15A T/C 0.79 56 924 −1.644 0.292 1.76E-08 0.81 76 913 −0.026 0.013 4.93E-02 0.81 −0.029 0.013 2.66E-02 HDL-C 17:41 840 849 s77697917 SOST-DUSP3 T/C 0.02 45 052 −2.717 0.407 2.38E-11 0.03 67 843 −0.222 0.036 4.27E-10 0.03 −0.241 0.035 1.04E-11 TG 19:8 429 323 s116843064 ANGPTL4 A/G 0.03 35 643 −0.101 0.016 6.46E-11 0.03 44 194 −0.065 0.019 4.53E-04 0.03 −0.087 0.012 3.83E-13 TC 20:17 844 684 s2618566 BANF2-SNX5 T/G 0.65 63 300 −1.566 0.251 4.68E-10 0.60 88 946 −0.024 0.011 2.83E-02 0.60 −0.027 0.011 1.38E-02 The a ian s in bold a e he signi ican ly eplica ed a ian s. A1 is allele 1 and A2 is allele 2, F eq is he equency o A1, βis he e ec o A1. HDL-C, high-densi y lipop o ein choles e ol; LDL-C, low-densi y lipop o ein choles e ol; TC, o al choles e ol; TG, iglyce ides. 444 an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om Bo h a ian s only pass quali y con ol in he coho s in he disco e y s age ha con ain indi iduals o A ican ances y (see online supplemen a y figu e S7). Al hough he e a e se e al coho s wi h indi iduals o A ican ances y in he eplica ion s age, bo h a ian s did no pass quali y con ol in mos coho s which leads o he conclusion ha hese a ian s migh be popula ion-specific. This is also sugges ed by he 1 kG da a (Phase 3) as he equency o he C-allele is 92% in A ican samples and 100% in he Eu opean samples o s60839105 and he equency o he G-allele is 86% in he A ican samples and 100% in he Eu opean samples o s151198427. Impu a ions o coho s wi h indi iduals o A ican ances y wi h he A ican Genome Va ia ion P ojec 20 migh confi m he associa ion o s60839105 wi h HDL-C and s151198427 wi h TC. To ou knowledge, his is he fi s GWAS o ci cula ing lipid le els using he Phase 1 in eg a ed elease V.3 o he 1 kG, he e- o e we canno compa e he posi i e eplica ion a e wi h o he s udies. Howe e , we did eplica e 88.1% o he findings o Teslo ich e al 2 and 43.4% o he findings o GLGC 3 despi e ou smalle sample. A high eplica ion a e is expec ed based on he high o e lap o ou samples wi h he samples o Teslo ich e al 2 and wi h he samples o GLGC 3 hough i indica es ha when using he 1000 Genomes ins ead o he HapMap e e - ence panel, we can achie e a high eplica ion a e using a smalle sample size. We also ied o eplica e findings om Figu e 2 The egional associa ion esul s o he ini ial me a-analysis o all disco e y coho s o (A) TC on ch omosome 6, (B) HDL-C on ch omosome 17, (C) TG on ch omosome 19, (D) LDL-C on ch omosome 6 and (E) TC on ch omosome 6. HDL-C, high-densi y lipop o ein choles e ol; LDL-C, low-densi y lipop o ein choles e ol; TC, o al choles e ol; TG, iglyce ides. an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 445 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om exome sequencing o candida e genes. The p.A g406X mu a ion in he NPC1L1 gene ( s145297799), which was epo ed o be associa ed wi h educed LDL-C le els and educed isk o co - ona y hea disease, 4 is no a ailable in he 1kG e e ence panel and, he e o e, we we e no able o eplica e his finding. Do e al 5 desc ibed he exome sequencing o he genes LDLR and APOA5 and iden ified a e a ian s associa ed wi h an inc eased isk o myoca dial in a c ion, inc eased LDL-C and TG le els. O hose a e a ian s, only wo in he LDLR gene and se en in he APOA5 gene exis in ou disco e y me a-analysis. Bo h LDLR a ian s a e associa ed wi h TG in ou disco e y me a-analysis ( s34282181, β=−0.093, SE β =0.023, p alue=4.827×10 −5 and s2075291, β=0.219, SE β =0.046, p alue=2.092×10 −6 ), bu no significan ly associa ed wi h LDL-C ( s34282181, β=−3.939, SE β =1.861, p alue=0.034 and s2075291, β=−2.316, SE β =3.001, p alue=0.440). None o he se en APOA5 a ian s we e significan ly associa ed wi h TG o LDL-C in ou disco e y me a-analysis (lowes p alue is o LDL-C wi h s72658860, β=−18.430, SE β =7.140, p alue=9.848×10 −3 ). The hi d published finding we ied o eplica e, was he associa ion be ween APOC3 and TG le els. 6 O he se en a ian s epo ed, only one exis ed in ou disco - e y me a-analysis (ch omosome 11, posi ion 116 701 354), which is associa ed wi h TG (β=−0.343, SE β =0.113, p alue=2.311×10 −3 ). Those au ho s also epo ed an associa ion be ween an APOA5 a ian ( s3135506) and TG as he mos sig- nifican finding. This a ian was also significan ly associa ed wi h TG in ou disco e y me a-analysis (β=0.129, SE β =0.007, p alue=1.099×10 −87 ). These eplica ion e o s demons a e ha many o he published esul s o exome sequencing can be eplica ed h ough he use o 1 kG impu a ions. In conclusion, we iden ified and eplica ed fi e a ian s asso- cia ed wi h ci cula ing lipid le els. These a ian s a e in genes ha can be linked biologically o lipid me abolism. Al hough he e we e a la ge numbe o a ian s ha did no eplica e a he accep ed genome-wide significance h eshold, he low-cos , hypo hesis- ee app oach ha we applied unco e ed fi e a - ian s. This s udy, he e o e, illus a es ha GWAS may s ill help us un a el he biological mechanisms behind ci cula ing lipid le els. Au ho a filia ions 1 Depa men o Epidemiology, E asmus Medical Cen e , Ro e dam, The Ne he lands 2 Human Genome Sequencing Cen e , Baylo College o Medicine, Hous on, USA 3 Depa men o Medicine, Uni e si y o Washing on, Sea le, USA 4 Medical Resea ch Council Human Gene ics Uni , Ins i u e o Gene ics and Molecula Medicine, Uni e si y o Edinbu gh, Edinbu gh, UK 5 The F amingham Hea S udy, NHLBI Ca dio ascula Epidemiology and Human Genomics B anch, F amingham, USA 6 Cen e o Public Heal h Genomics, Uni e si y o Vi ginia, Cha lo es ille, USA 7 Icelandic Hea Associa ion, Kopa ogu , Iceland 8 Facul y o Medicine, Uni e si y o Iceland, Reykja ik, Iceland 9 Depa men o Gene ics, Washing on Uni e si y School o Medicine, S Louis, USA 10 Depa men o Bios a is ics, Bos on Uni e si y School o Public Heal h, Bos on, USA 11 Ushe Ins i u e o Popula ion Heal h Sciences and In o ma ics, Uni e si y o Edinbu gh, Edinbu gh, UK 12 Depa men o Gene ics, Uni e si y o No h Ca olina, Chapel Hill, USA 13 Depa men o Human Gene ics, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 14 Human Genomics Uni , Ins i u e o Molecula Medicine, Uni e si y o Helsinki, Helsinki, Finland 15 Depa men o Epidemiology, Uni e si y o G oningen, Uni e si y Medical Cen e G oningen, G oningen, The Ne he lands 16 Depa men o Epidemiology and Bios a is ics, School o Public Heal h, Impe ial College London, London, UK 17 Depa men o Ca diology, Ealing Hospi al NHS T us , Middlesex, UK 18 Depa men o Biological Psychology, VU Uni e si y Ams e dam and EMGO+ Ins i u e o Heal h and Ca e Resea ch, Ams e dam, The Ne he lands 19 Depa men o Clinical Epidemiology, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 20 Depa men o Ca diology, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 21 Depa men o Ge on ology and Ge ia ics, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 22 Depa men o Ca diology, Uni e si y Medical Cen e G oningen, Uni e si y o G oningen, G oningen, The Ne he lands 23 Depa men o Hygiene and Epidemiology, Uni e si y o Ioannina Medical School, Ioannina, G eece 24 Depa men o Clinical Chemis y, Fimlab Labo a o ies, Tampe e, Finland 25 Depa men o Clinical Chemis y, Uni e si y o Tampe e School o Medicine, Tampe e, Finland 26 Public Heal h Sciences, Loyola Uni e si y Chicago S i ch School o Medicine, Maywood, USA 27 Depa men o Molecula Epidemiology, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 28 Labo a o y o Expe imen al Ca diology, Uni e si y Medical Cen e U ech , U ech , The Ne he lands 29 McKusick-Na hans Ins i u e o Gene ic Medicine, Johns Hopkins Uni e si y School o Medicine, Bal imo e, USA 30 Human Gene ics Cen e , The Uni e si y o Texas School o Public Heal h, Hous on, USA 31 Depa men o In e nal Medicine, E asmus Medical Cen e , Ro e dam, The Ne he lands 32 Gene a ion Sco land, Cen e o Genomic and Expe imen al Medicine, Ins i u e o Gene ics and Molecula Medicine, Uni e si y o Edinbu gh, Edinbu gh, UK 33 Musculoskele al Resea ch P og amme, Di ision o Applied Medicine, Uni e si y o Abe deen, Abe deen, UK 34 B i ish Hea Founda ion Glasgow Ca dio ascula Resea ch Cen e, Ins i u e o Ca dio ascula and Medical Sciences, College o Medical, Ve e ina y and Li e Sciences, Uni e si y o Glasgow, Glasgow, UK 35 Depa men o Bios a is ics, Uni e si y o Washing on, Sea le, USA 36 B igham and Women’s Hospi al, Bos on, USA 37 Labo a o y o Epidemiology and Popula ion Sciences, Na ional Ins i u e on Aging, Na ional Ins i u es o Heal h, Be hesda, USA 38 Cen e o Popula ion Heal h Sciences, Uni e si y o Edinbu gh, Edinbu gh, UK 39 Facul y o Medicine, Uni e si y o Spli , Spli , C oa ia 40 Depa men o Heal h, Na ional Ins i u e o Heal h and Wel a e, Helsinki, Finland 41 Ca dio ascula Science, Na ional Hea and Lung Ins i u e, Impe ial College London, London, UK 42 Impe ial College Heal hca e NHS T us, Impe ial College London, London, UK 43 Depa men o Psychia y, VU Uni e si y Medical Cen e Ams e dam/GGZinGees and EMGO+ Ins i u e o Heal h and Ca e Resea ch and Neu oscience Campus Ams e dam, Ams e dam, The Ne he lands 44 Robe son Cen e o Bios a is ics, Uni e si y o Glasgow, Glasgow, UK 45 Depa men o Neph ology, Uni e si y o G oningen, Uni e si y Medical Cen e G oningen, G oningen, The Ne he lands 46 Depa men o Clinical Physiology, Tu ku Uni e si y Hospi al, Tu ku, Finland 47 Resea ch Cen e o Applied and P e en i e Ca dio ascula Medicine, Uni e si y o Tu ku, Tu ku, Finland 48 Di ision o Medicine, Tu ku Uni e si y Hospi al, Tu ku, Finland 49 Depa men o Medicine, Uni e si y o Tu ku, Tu ku, Finland 50 Depa men o Ca diology, Hea Hospi al, Tampe e Uni e si y Hospi al, Tampe e, Finland 51 School o Medicine, Uni e si y o Tampe e, Tampe e, Finland 52 T opical Me abolism Resea ch Uni , T opical Medicine Resea ch Ins i u e, Uni e si y o he Wes Indies, Mona, Jamaica 53 Labo a o y o Clinical Chemis y and Hema ology, Uni e si y Medical Cen e U ech , U ech , The Ne he lands 54 Depa men o Psychia y, Uni e si y o G oningen, Uni e si y Medical Cen e G oningen, G oningen, The Ne he lands 55 Depa men o Clinical Physiology, Tampe e Uni e si y Hospi al, Tampe e, Finland 56 Depa men o Clinical Physiology, Uni e si y o Tampe e School o Medicine, Tampe e, Finland 57 Depa men o Epidemiology and Bios a is ics, MRC-PHE Cen e o En i onmen and Heal h, School o Public Heal h, Impe ial College London, London, UK 58 Depa men o Gene ics, Uni e si y o G oningen, Uni e si y Medical Cen e G oningen, G oningen, The Ne he lands 59 Depa men o Public Heal h and P ima y Ca e, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 60 Epidemiology Sec ion, Depa men o BESC, King Faisal Medical Hospi al and Resea ch Cen e, Riyadh, Saudi A abia 61 Genomics Coo dina ion Cen e , Uni e si y o G oningen, Uni e si y Medical Cen e G oningen, G oningen, The Ne he lands 62 Public Heal h, Uni e si y o Helsinki, Helsinki, Finland 63 Wellcome T us Sange Ins i u e, UK 446 an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om 64 Depa men o Gene al In e nal Medicine, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands 65 Physiology and Biophysics, Uni e si y o Mississippi Medical Cen e , Jackson, USA 66 Depa men o Epidemiology, Uni e si y o Washing on, Sea le, USA 67 Depa men o Heal h Se ices, Uni e si y o Washing on, Sea le, USA 68 G oup Heal h Coope a i e, G oup Heal h Resea ch Ins i u e, Sea le, USA 69 Ins i u e o T ansla ional Genomics and Popula ion Sciences, Los Angeles BioMedical Resea ch Ins i u e a Ha bo -UCLA Medical Cen e , To ance, USA 70 Di ision o Genomic Ou comes, Depa men o Pedia ics, Ha bo -UCLA Medical Cen e , To ance, USA 71 Depa men s o Pedia ics, Medicine, and Human Gene ics, UCLA, Los Angeles, USA Acknowledgemen s The au ho s especially hank all olun ee s who pa icipa ed in he s udy. The au ho s hank he s a and pa icipan s o he ARIC s udy o hei impo an con ibu ions. In as uc u e was pa ly suppo ed by G an Numbe UL1RR025005, a componen o he Na ional Ins i u es o Heal h and NIH Roadmap o Medical Resea ch. We a e g a e ul o all s udy pa icipan s and hei ela i es, gene al p ac i ione s and neu ologis s o hei con ibu ions o he ERF s udy and o P Ve aa o he help in genealogy, J Ve gee o he supe ision o he labo a o y wo k and P Snijde s o his help in da a collec ion. The au ho s hank Beh ooz Alizadeh, Annemieke Boesjes, Ma cel B uinenbe g, Noo je Fes en, Pim an de Ha s , Ilja Nol e, Lude F anke, Mi a Valimohammadi o hei help in c ea ing he GWAS da abase, and Rob Bie inga, Joos Kee s, René Oos e go, Rosalie Visse , Judi h Vonk o hei wo k ela ed o da a collec ion and alida ion. The au ho s a e g a e ul o he s udy pa icipan s, he s a om he Li eLines Coho S udy and he con ibu ing esea ch cen es deli e ing da a o Li eLines and he pa icipa ing gene al p ac i ione s and pha macis s. MESA and he MESA SHARe p ojec a e conduc ed and suppo ed by con ac s N01-HC-95159, N01-HC-95160, N01-HC-95161, N01-HC-95162, N01-HC-95163, N01-HC-95164, N01-HC-95165, N01-HC-95166, N01-HC-95167, N01-HC-95168, N01-HC-95169, UL1-TR-001079 and UL1-TR-000040 om he Na ional Hea , Lung, and Blood Ins i u e (NHLBI). Funding o MESA SHARe geno yping was p o ided by NHLBI Con ac N02-HL6-4278. The p o ision o geno yping da a was suppo ed in pa by he Na ional Cen e o Ad ancing T ansla ional Sciences, CTSI g an UL1TR000124, and he Na ional Ins i u e o Diabe es and Diges i e and Kidney Disease Diabe es Resea ch Cen e (DRC) g an DK063491 o he Sou he n Cali o nia Diabe es Endoc inology Resea ch Cen e . This publica ion was de eloped unde a STAR esea ch assis ance ag eemen , No. RD831697 (MESA Ai ), awa ded by he U.S En i onmen al p o ec ion Agency. I has no been o mally e iewed by he EPA. The iews exp essed in his documen a e solely hose o he au ho s and he EPA does no endo se any p oduc s o comme cial se ices men ioned in his publica ion. The au ho s o he NEO s udy hank all indi iduals who pa icipa ed in he Ne he lands Epidemiology in Obesi y s udy, all pa icipa ing gene al p ac i ione s o in i ing eligible pa icipan s and all esea ch nu ses o collec ion o he da a. The au ho s hank he NEO s udy g oup, Pe a Noo dijk, Pa an Beelen and Ingebo g de Jonge o he coo dina ion, lab and da a managemen o he NEO s udy. The geno yping in he NEO s udy was pe o med a he Cen e Na ional de Géno ypage (Pa is, F ance), headed by Jean-F ancois Deleuze. The au ho s o he ORCADES s udy would like o acknowledge he in aluable con ibu ions o Lo aine Ande son and he esea ch nu ses in O kney, he adminis a i e eam in Edinbu gh and he people o O kney. On behal o he Ro e dam S udy, he au ho s hank Pascal A p, Mila Jhamai, Ma ijn Ve ke k, Lizbe h He e a and Ma jolein Pe e s o hei help in c ea ing he GWAS da abase, and Ka ol Es ada and Maksim V. S uchalin o hei suppo in c ea ion and analysis o impu ed da a. The au ho s a e g a e ul o he s udy pa icipan s, he s a om he Ro e dam S udy and he pa icipa ing gene al p ac i ione s and pha macis s. Collabo a o s The Li eLines Coho S udy: see online supplemen a y appendix 1. CHARGE Lipids Wo king G oup: see online supplemen a y appendix 2. Con ibu o s EM L o ganised he s udy and designed he s udy wi h subs an ial inpu o AI, LAC and CM D. EM L d a ed he manusc ip wi h subs an ial inpu om CM D. All au ho s had he oppo uni y o commen on he manusc ip . Da a collec ion, GWAS and s a is ical analysis was done by SW dL, HMdR, GP (AEGS); AVS, VG, TBH (AGES); EE, MPS, PE (Ai wa e); AS, DEA, ACM, EB (ARIC); JCB, JAB, KMC, BMP (CHS); AI, EM L, CM D (ERF); MFF, IBB (FamHS); LPL, KN, MK (FINCAVAS); IS, VS, SR (FINRISK); SD, CCW, LAC (FHS); JEH, AC, LJH, SP (GS); QD, LAL, JGW (JHS); JEH, IK, PN, OP (CROATIA Ko cula); IMN, MS (Li elines); JD, AJMdC, PES (LLS); WZ, JSK, BL, WRS, STT, JCC (LOLIPOP); BOT, TF, CAM, RSC (Loyola); AM, JCM, SSR, JIR (MESA); RLG, RdM, DOMK (NEO); HM, EJdG, YM, BWJHP, GW, DIB (NTR-NESDA); PKJ, HC, JFW (ORCADES); NV, RTG, P dH (PREVEND); ST, IF, JWJ (PROSPER); EM L, AD, OHF, AH, FR, EJS, AGU, CM D (RS); JEH, TZ, VV (CROATIA Spli ); PJ dM, AJO, HS (TRAILS); JEH, JM, CH, IR (CROATIA Vis); JSV, OTR, TL (YFS). EM L pe o med he me a-analysis and all ollow-up s eps. Biological associa ion o loci and bioin o ma ics we e ca ied ou by EM L, JB K, KW D and CM D. Funding The AGES S udy has been unded by NIH con ac s N01-AG-1-2100 and HHSN271201200022C, he NIA In amu al Resea ch P og am, Hja a e nd ( he Icelandic Hea Associa ion), and he Al hingi ( he Icelandic Pa liamen ). The Ai wa e S udy is unded by he Home O fice (g an numbe 780-TETRA) wi h addi ional suppo om he Na ional Ins i u e o Heal h Resea ch (NIHR) Impe ial College Heal hca e NHS T us (ICHNT) and Impe ial College Biomedical Resea ch Cen e (BRC). PE is an NIHR Senio In es iga o and is suppo ed by he ICHNT and Impe ial College BRC, he MRC-PHE Cen e o En i onmen and Heal h and he NIHR Heal h P o ec ion Resea ch Uni on Heal h Impac o En i onmen al Haza ds. The ARIC S udy is ca ied ou as a collabo a i e s udy suppo ed by Na ional Hea , Lung, and Blood Ins i u e (NHLBI) con ac s (HHSN268201100005C, HHSN268201100006C, HHSN268201100007C, HHSN268201100008C, HHSN268201100009C, HHSN268201100010C, HHSN268201100011C, and HHSN268201100012C), R01HL087641, R01HL59367 and R01HL086694; Na ional Human Genome Resea ch Ins i u e con ac U01HG004402; and Na ional Ins i u es o Heal h con ac HHSN268200625226C. Ca dio ascula Heal h S udy: This CHS esea ch was suppo ed by NHLBI con ac s HHSN268201200036C, HHSN268200800007C, N01HC55222, N01HC85079, N01HC85080, N01HC85081, N01HC85082, N01HC85083, N01HC85086; and NHLBI g an s U01HL080295, R01HL087652, R01HL105756, R01HL103612, and R01HL120393 wi h addi ional con ibu ion om he Na ional Ins i u e o Neu ological Diso de s and S oke (NINDS). Addi ional suppo was p o ided h ough R01AG023629 om he Na ional Ins i u e on Aging (NIA). A ull lis o p incipal CHS in es iga o s and ins i u ions can be ound a CHS-NHLBI.o g. The p o ision o geno yping da a was suppo ed in pa by he Na ional Cen e o Ad ancing T ansla ional Sciences, CTSI g an UL1TR000124, and he Na ional Ins i u e o Diabe es and Diges i e and Kidney Disease Diabe es Resea ch Cen e (DRC) g an DK063491 o he Sou he n Cali o nia Diabe es Endoc inology Resea ch Cen e . The con en is solely he esponsibili y o he au ho s and does no necessa ily ep esen he o ficial iews o he Na ional Ins i u es o Heal h. CROATIA-Ko cula, CROATIA-Spli and CROATIA-Vis (CR-Ko cula, CR-Spli , CR-Vis) we e unded by he Medical Resea ch Council UK, The C oa ian Minis y o Science, Educa ion and Spo s (g an 216-1080315-0302), he Eu opean Union amewo k p og am 6 EUROSPAN p ojec (con ac no. LSHG-CT-2006-018947) and he C oa ian Science Founda ion (g an 8875). The ERF s udy as a pa o EUROSPAN (Eu opean Special Popula ions Resea ch Ne wo k) was suppo ed by Eu opean Commission FP6 STRP g an numbe 018947 (LSHG-CT-2006-01947) and also ecei ed unding om he Eu opean Communi y’s Se en h F amewo k P og amme (FP7/2007-2013)/g an ag eemen HEALTH-F4-2007-201413 by he Eu opean Commission unde he p og amme “Quali y o Li e and Managemen o he Li ing Resou ces”o 5 h F amewo k P og amme (no. QLG2-CT-2002-01254). The ERF s udy was u he suppo ed by ENGAGE conso ium and CMSB. High- h oughpu analysis o he ERF da a was suppo ed by join g an om Ne he lands O ganisa ion o Scien ific Resea ch and he Russian Founda ion o Basic Resea ch (NWO-RFBR 047.017.043). Exome sequencing in ERF was suppo ed by he ZonMw g an (p ojec 91111025). The Family Hea S udy was suppo ed by he by g an s R01-HL-087700, R01-HL-088215 and R01-HL-117078 om he Na ional Hea , Lung, and Blood Ins i u e. Gene a ion Sco land ecei ed co e unding om he Chie Scien is O fice o he Sco ish Go e nmen Heal h Di ec o a e CZD/16/6 and he Sco ish Funding Council HR03006. Geno yping o he GS:SFHS samples was ca ied ou by he Gene ics Co e Labo a o y a he Wellcome T us Clinical Resea ch Facili y, Edinbu gh, Sco land and was unded by he UK’s Medical Resea ch Council. The Li eLines Coho S udy, and gene a ion and managemen o GWAS geno ype da a o he Li eLines Coho S udy is suppo ed by he Ne he lands O ganiza ion o Scien ific Resea ch NWO (g an 175.010.2007.006), he Economic S uc u e Enhancing Fund (FES) o he Du ch go e nmen , he Minis y o Economic A ai s, he Minis y o Educa ion, Cul u e and Science, he Minis y o Heal h, Wel a e and Spo s, he No he n Ne he lands Collabo a ion o P o inces (SNN), he P o ince o G oningen, Uni e si y Medical Cen e G oningen, he Uni e si y o G oningen, Du ch Kidney Founda ion and Du ch Diabe es Resea ch Founda ion. The Leiden Longe i y S udy (LLS) has ecei ed unding om he Eu opean Union’s Se en h F amewo k P og amme (FP7/2007–2011) unde g an ag eemen no 259679. This s udy was suppo ed by a g an om he Inno a ion-O ien ed Resea ch P og am on Genomics (Sen e No em IGE05007), he Cen e o Medical Sys ems Biology, and he Ne he lands Conso ium o Heal hy Ageing (g an 050-060-810), all in he amewo k o he Ne he lands Genomics Ini ia i e, Ne he lands O ganiza ion o Scien ific Resea ch (NWO), Unile e Colwo h and by BBMRI-NL, a Resea ch In as uc u e financed by he Du ch go e nmen (NWO 184.021.007). The LOLIPOP s udy is suppo ed by he Na ional Ins i u e o Heal h Resea ch (NIHR) Comp ehensi e Biomedical Resea ch Cen e Impe ial College Heal hca e NHS T us , he B i ish Hea Founda ion (SP/04/002), he Medical Resea ch Council (G0601966,G0700931), he Wellcome T us (084723/Z/08/Z) he NIHR an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 447 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om (RP-PG-0407-10371),Eu opean Union FP7 (EpiMig an , 279143) and Ac ion on Hea ing Loss (G51). The wo k was ca ied ou in pa a he NIHR/Wellcome T us Impe ial Clinical Resea ch Facili y. The NEO s udy is suppo ed by he pa icipa ing Depa men s, he Di ision and he Boa d o Di ec o s o he Leiden Uni e si y Medical Cen e , and by he Leiden Uni e si y, Resea ch P ofile A ea ‘Vascula and Regene a i e Medicine’. Dennis Mook-Kanamo i is suppo ed by Du ch Science O ganiza ion (ZonMW-VENI G an 916.14.023). ORCADES was suppo ed by he Chie Scien is O fice o he Sco ish Go e nmen , he Royal Socie y, he MRC Human Gene ics Uni , A h i is Resea ch UK and he Eu opean Union amewo k p og am 6 EUROSPAN p ojec (con ac no. LSHG-CT-2006-018947). DNA ex ac ions we e pe o med a he Wellcome T us Clinical Resea ch Facili y in Edinbu gh. The PROSPER s udy was suppo ed by an in es iga o ini ia ed g an ob ained om B is ol-Mye s Squibb. JWJ is an Es ablished Clinical In es iga o o he Ne he lands Hea Founda ion (g an 2001 D 032). Suppo o geno yping was p o ided by he se en h amewo k p og am o he Eu opean commission (g an 223004) and by he Ne he lands Genomics Ini ia i e (Ne he lands Conso ium o Heal hy Aging g an 050-060-810). The gene a ion and managemen o GWAS geno ype da a o he Ro e dam S udy is suppo ed by he Ne he lands O ganisa ion o Scien ific Resea ch NWO In es men s (n . 175.010.2005.011, 911-03-012). This s udy is unded by he Resea ch Ins i u e o Diseases in he Elde ly (014-93-015; RIDE2), he Ne he lands Genomics Ini ia i e (NGI)/Ne he lands O ganisa ion o Scien ific Resea ch (NWO) p ojec n . 050-060-810. The Ro e dam S udy is unded by E asmus Medical Cen e and E asmus Uni e si y, Ro e dam, Ne he lands O ganiza ion o he Heal h Resea ch and De elopmen (ZonMw), he Resea ch Ins i u e o Diseases in he Elde ly (RIDE), he Minis y o Educa ion, Cul u e and Science, he Minis y o Heal h, Wel a e and Spo s, he Eu opean Commission (DG XII), and he Municipali y o Ro e dam. Pa icipa ing cen es o TRAILS include he Uni e si y Medical Cen e and Uni e si y o G oningen, he E asmus Uni e si y Medical Cen e Ro e dam, he Uni e si y o U ech , he Radboud Medical Cen e Nijmegen, and he Pa nassia Ba o g oup, all in he Ne he lands. TRAILS has been financially suppo ed by a ious g an s om he Ne he lands O ganiza ion o Scien ific Resea ch NWO (Medical Resea ch Council p og am g an GB-MW 940-38-011; ZonMW B ainpowe g an 100-001-004; ZonMw Risk Beha io and Dependence g an s 60-60600-97-118; ZonMw Cul u e and Heal h g an 261-98-710; Social Sciences Council medium-sized in es men g an s GB-MaGW 480-01-006 and GB-MaGW 480-07-001; Social Sciences Council p ojec g an s GB-MaGW 452-04-314 and GB-MaGW 452-06-004; NWO la ge-sized in es men g an 175.010.2003.005; NWO Longi udinal Su ey and Panel Funding 481-08-013 and 481-11-001), he Du ch Minis y o Jus ice (WODC), he Eu opean Science Founda ion (Eu oSTRESS p ojec FP-006), Biobanking and Biomolecula Resou ces Resea ch In as uc u e BBMRI-NL (CP 32), and he pa icipa ing uni e si ies. Compe ing in e es s BMP se es on he DSMB o a clinical ial o a de ice unded by he manu ac u e (Zoll Li eCo ) and on he S ee ing Commi ee o he Yale Open Da a Access P ojec unded by Johnson & Johnson. OHF wo ks in E asmusAGE, a cen e o aging esea ch ac oss he li e cou se unded by Nes lé Nu i ion (Nes ec); Me agenics; and AXA. Nes lé Nu i ion (Nes ec); Me agenics; and AXA had no ole in design and conduc o he s udy; collec ion, managemen , analysis, and in e p e a ion o he da a; and p epa a ion, e iew o app o al o he manusc ip . Pa ien consen Ob ained. E hics app o al All s udies we e pe o med wi h he app o al o he Local Medical E hics Commi ees. P o enance and pee e iew No commissioned; ex e nally pee e iewed. Da a sha ing s a emen All me a-analysis esul s and he esul s o he ollow-up analysis o his p ojec a e a ailable. Open Access This is an Open Access a icle dis ibu ed in acco dance wi h he C ea i e Commons A ibu ion Non Comme cial (CC BY-NC 4.0) license, which pe mi s o he s o dis ibu e, emix, adap , build upon his wo k non-comme cially, and license hei de i a i e wo ks on di e en e ms, p o ided he o iginal wo k is p ope ly ci ed and he use is non-comme cial. See: h p://c ea i ecommons.o g/ licenses/by-nc/4.0/ REFERENCES 1 Su akka I, Ho ikoshi M, Mägi R, Sa in AP, Mahajan A, Lagou V, Ma ullo L, Fe ei a T, Mi aglio B, Timonen S, Ke unen J, Pi inen M, Ka jalainen J, Tho lei sson G, Hägg S, Ho enga JJ, Isaacs A, Laden all C, Beekman M, Esko T, Ried JS, Nelson CP, Willenbo g C, Gus a sson S, Wes a HJ, Blades M, de C aen AJ, de Geus EJ, Deelen J, G alle H, Hams en A, Ha ulinna AS, Hengs enbe g C, Houwing-Duis e maa JJ, Hyppönen E, Ka ssen LC, Leh imäki T, Lyssenko V, Magnusson PK, Mihailo E, Mülle -Nu asyid M, Mpindi JP, Pede sen NL, Penninx BW, Pe ola M, Pe s TH, Pe e s A, Rung J, Smi JH, S ein ho sdo i V, Tobin MD, Tse niko a N, an Leeuwen EM, Viika i JS, Willems SM, Willemsen G, Schunke H, E dmann J, Samani NJ, Kap io J, Lind L, Giege C, Me spalu A, Slagboom PE, G oop L, an Duijn CM, E iksson JG, Jula A, Salomaa V, Boomsma DI, Powe C, Rai aka i OT, Ingelsson E, Jä elin MR, Tho s einsdo i U, F anke L, Ikonen E, Kallioniemi O, Pie iäinen V, Lindg en CM, S e ansson K, Palo ie A, McCa hy MI, Mo is AP, P okopenko I, Ripa i S, ENGAGE Conso ium. The impac o low- equency and a e a ian s on lipid le els. Na Gene 2015;47:589–97. 2 Teslo ich TM, Musunu u K, Smi h AV, Edmondson AC, S ylianou IM, Koseki M, Pi uccello JP, Ripa i S, Chasman DI, Wille CJ, Johansen CT, Fouchie SW, Isaacs A, Peloso GM, Ba balic M, Ricke s SL, Bis JC, Aulchenko YS, Tho lei sson G, Fei osa MF, Chambe s J, O ho-Melande M, Melande O, Johnson T, Li X, Guo X, Li M, Shin Cho Y, Jin Go M, Jin Kim Y, Lee JY, Pa k T, Kim K, Sim X, Twee-Hee Ong R, C o eau-Chonka DC, Lange LA, Smi h JD, Song K, Hua Zhao J, Yuan X, Luan J, Lamina C, Ziegle A, Zhang W, Zee RY, W igh AF, Wi eman JC, Wilson JF, Willemsen G, Wichmann HE, Whi field JB, Wa e wo h DM, Wa eham NJ, Waebe G, Vollenweide P, Voigh BF, Vi a V, Ui e linden AG, Uda M, Tuomileh o J, Thompson JR, Tanaka T, Su akka I, S ingham HM, Spec o TD, So anzo N, Smi JH, Sinisalo J, Silande K, Sijb ands EJ, Scu e i A, Sco J, Schlessinge D, Sanna S, Salomaa V, Saha inen J, Saba i C, Ruokonen A, Rudan I, Rose LM, Robe s R, Riede M, Psa y BM, P ams alle PP, Pichle I, Pe ola M, Penninx BW, Pede sen NL, Pa a o C, Pa ke AN, Pa e G, Oos a BA, O’Donnell CJ, Nieminen MS, Nicke son DA, Mon gome y GW, Mei inge T, McPhe son R, McCa hy MI, McA dle W, Masson D, Ma in NG, Ma oni F, Mangino M, Magnusson PK, Lucas G, Luben R, Loos RJ, Lokki ML, Le e G, Langenbe g C, Laune LJ, Laka a EG, Laaksonen R, Ky ik KO, K onenbe g F, König IR, Khaw KT, Kap io J, Kaplan LM, Johansson A, Ja elin MR, Janssens AC, Ingelsson E, Igl W, Kees Ho ingh G, Ho enga JJ, Ho man A, Hicks AA, Hengs enbe g C, Heid IM, Haywa d C, Ha ulinna AS, Has ie ND, Ha is TB, Ha i unians T, Hall AS, Gyllens en U, Guiducci C, G oop LC, Gonzalez E, Giege C, F eime NB, Fe ucci L, E dmann J, Ellio P, Ejebe KG, Dö ing A, Dominiczak AF, Demissie S, Deloukas P, de Geus EJ, de Fai e U, C aw o d G, Collins FS, Chen YD, Caulfield MJ, Campbell H, Bu NP, Bonnycas le LL, Boomsma DI, Boekhold SM, Be gman RN, Ba oso I, Bandinelli S, Ballan yne CM, Assimes TL, Que e mous T, Al shule D, Seiels ad M, Wong TY, Tai ES, Fe anil AB, Kuzawa CW, Adai LS, Taylo HA J , Bo ecki IB, Gab iel SB, Wilson JG, Holm H, Tho s einsdo i U, Gudnason V, K auss RM, Mohlke KL, O do as JM, Mun oe PB, Koone JS, Tall AR, Hegele RA, Kas elein JJ, Schad EE, Ro e JI, Boe winkle E, S achan DP, Moose V, S e ansson K, Reilly MP, Samani NJ, Schunke H, Cupples LA, Sandhu MS, Ridke PM, Rade DJ, an Duijn CM, Pel onen L, Abecasis GR, Boehnke M, Ka hi esan S. Biological, clinical and popula ion ele ance o 95 loci o blood lipids. Na u e 2010;466:707–13. 3 Global Lipids Gene ics Conso iumWille CJ, Schmid EM, Sengup a S, Peloso GM, Gus a sson S, Kanoni S, Ganna A, Chen J, Buchko ich ML, Mo a S, Beckmann JS, B agg-G esham JL, Chang HY, Demi kan A, Den He og HM, Do R, Donnelly LA, Eh e GB, Esko T, Fei osa MF, Fe ei a T, Fische K, Fon anillas P, F ase RM, F ei ag DF, Gu dasani D, Heikkilä K, Hyppönen E, Isaacs A, Jackson AU, Johansson A, Johnson T, Kaakinen M, Ke unen J, Klebe ME, Li X, Luan J, Lyy ikäinen LP, Magnusson PK, Mangino M, Mihailo E, Mon asse ME, Mülle -Nu asyid M, Nol e IM, O’Connell JR, Palme CD, Pe ola M, Pe e sen AK, Sanna S, Saxena R, Se ice SK, Shah S, Shungin D, Sido e C, Song C, S awb idge RJ, Su akka I, Tanaka T, Teslo ich TM, Tho lei sson G, Van den He ik EG, Voigh BF, Volcik KA, Wai e LL, Wong A, Wu Y, Zhang W, Abshe D, Asiki G, Ba oso I, Been LF, Bol on JL, Bonnycas le LL, B ambilla P, Bu ne MS, Cesana G, Dimi iou M, Doney AS, Dö ing A, Ellio P, Eps ein SE, Eyjol sson GI, Gigan e B, Gooda zi MO, G alle H, G a i o ML, G o es CJ, Hallmans G, Ha ikainen AL, Haywa d C, He nandez D, Hicks AA, Holm H, Hung YJ, Illig T, Jones MR, Kaleebu P, Kas elein JJ, Khaw KT, Kim E, Klopp N, Komulainen P, Kuma i M, Langenbe g C, Leh imäki T, Lin SY, Linds öm J, Loos RJ, Mach F, McA dle WL, Meisinge C, Mi chell BD, Mülle G, Naga aja R, Na isu N, Nieminen TV, Nsubuga RN, Ola sson I, Ong KK, Palo ie A, Papama kou T, Pomilla C, Pou a A, Rade DJ, Reilly MP, Ridke PM, Ri adenei a F, Rudan I, Ruokonen A, Samani N, Scha nagl H, Seeley J, Silande K, S ancáko á A, S i ups K, Swi AJ, Ti e L, Ui e linden AG, an Pel LJ, Vedan am S, Wainw igh N, Wijmenga C, Wild SH, Willemsen G, Wilsgaa d T, Wilson JF, Young EH, Zhao JH, Adai LS, A eile D, Assimes TL, Bandinelli S, Benne F, Bochud M, Boehm BO, Boomsma DI, Bo ecki IB, Bo ns ein SR, Bo e P, Bu nie M, Campbell H, Chak a a i A, Chambe s JC, Chen YD, Collins FS, Coope RS, Danesh J, Dedoussis G, de Fai e U, Fe anil AB, Fe iè es J, Fe ucci L, F eime NB, Giege C, G oop LC, Gudnason V, Gyllens en U, Hams en A, Ha is TB, Hingo ani A, Hi schho n JN, Ho man A, Ho ingh GK, Hsiung CA, Humph ies SE, Hun SC, H eem K, I iba en C, Jä elin MR, Jula A, Kähönen M, Kap io J, Kesäniemi A, Ki imaki M, Koone JS, Kouds aal PJ, K auss RM, Kuh D, Kuusis o J, Ky ik KO, Laakso M, Lakka TA, Lind L, Lindg en CM, Ma in NG, Mä z W, McCa hy MI, McKenzie CA, Mene on P, Me spalu A, Moilanen L, Mo is AD, Mun oe PB, Njøls ad I, Pede sen NL, Powe C, P ams alle PP, P ice JF, Psa y BM, Que e mous T, Rau amaa R, Saleheen D, Salomaa V, Sanghe a DK, Sa amies J, Schwa z PE, Sheu WH, Shuldine AR, Siegbahn A, Spec o TD, S e ansson K, S achan DP, Tayo BO, T emoli E, Tuomileh o J, Uusi upa M, an Duijn CM, Vollenweide P, Wallen in L, Wa eham NJ, Whi field JB, Wol enbu el BH, O do as 448 an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om JM, Boe winkle E, Palme CN, Tho s einsdo i U, Chasman DI, Ro e JI, F anks PW, Ripa i S, Cupples LA, Sandhu MS, Rich SS, Boehnke M, Deloukas P, Ka hi esan S, Mohlke KL, Ingelsson E, Abecasis GR. Disco e y and efinemen o loci associa ed wi h lipid le els. Na Gene 2013;45:1274–83. 4 Myoca dial In a c ion Gene ics Conso ium In es iga o s. S i ziel NO, Won HH, Mo ison AC, Peloso GM, Do R, Lange LA, Fon anillas P, Gup a N, Duga S, Goel A, Fa all M, Saleheen D, Fe a io P, König I, Assel a R, Me lini PA, Ma ziliano N, No a angelo MF, Schick U, Aue P, Assimes TL, Reilly M, Wilensky R, Rade DJ, Ho ingh GK, Mei inge T, Kessle T, Kas a i A, Laugwi z KL, Sisco ick D, Ro e JI, Hazen SL, T acy R, C esci S, Spe us J, Jackson R, Schwa z SM, Na a ajan P, C osby J, Muzny D, Ballan yne C, Rich SS, O’Donnell CJ, Abecasis G, Sunyae S, Nicke son DA, Bu ing JE, Ridke PM, Chasman DI, Aus in E, Ye Z, Kullo IJ, Weeke PE, Sha e CM, Bas a ache LA, Denny JC, Roden DM, Palme C, Deloukas P, Lin DY, Tang ZZ, E dmann J, Schunke H, Danesh J, Ma uga J, Elosua R, A dissino D, McPhe son R, Wa kins H, Reine AP, Wilson JG, Al shule D, Gibbs RA, Lande ES, Boe winkle E, Gab iel S, Ka hi esan S. Inac i a ing mu a ions in NPC1L1 and p o ec ion om co ona y hea disease. N Engl J Med 2014;371:2072–82. 5 Do R, S i ziel NO, Won HH, Jø gensen AB, Duga S, Angelica Me lini P, Kiezun A, Fa all M, Goel A, Zuk O, Guella I, Assel a R, Lange LA, Peloso GM, Aue PL; NHLBI Exome Sequencing P ojec , Gi elli D, Ma inelli N, Fa low DN, DeP is o MA, Robe s R, S ewa AF, Saleheen D, Danesh J, Eps ein SE, Si apala a nam S, Ho ingh GK, Kas elein JJ, Samani NJ, Schunke H, E dmann J, Shah SH, K aus WE, Da ies R, Nikpay M, Johansen CT, Wang J, Hegele RA, Hech e E, Ma z W, Klebe ME, Huang J, Johnson AD, Li M, Bu ke GL, G oss M, Liu Y, Assimes TL, Heiss G, Lange EM, Folsom AR, Taylo HA, Oli ie i O, Hams en A, Cla ke R, Reilly DF, Yin W, Ri as MA, Donnelly P, Rossouw JE, Psa y BM, He ing on DM, Wilson JG, Rich SS, Bamshad MJ, T acy RP, Cupples LA, Rade DJ, Reilly MP, Spe us JA, C esci S, Ha iala J, Tang WH, Hazen SL, Allayee H, Reine AP, Ca lson CS, Koope be g C, Jackson RD, Boe winkle E, Lande ES, Schwa z SM, Sisco ick DS, McPhe son R, Tybjae g-Hansen A, Abecasis GR, Wa kins H, Nicke son DA, A dissino D, Sunyae SR, O’Donnell CJ, Al shule D, Gab iel S, Ka hi esan S. Exome sequencing iden ifies a e LDLR and APOA5 alleles con e ing isk o myoca dial in a c ion. Na u e 2015;518:102–6. 6 The TG and HDL Wo king G oup o he Exome Sequencing P ojec , Na ional Hea , Lung, and Blood Ins i u eC osby J, Peloso GM, Aue PL, C osslin DR, S i ziel NO, Lange LA, Lu Y, Tang ZZ, Zhang H, Hindy G, Masca N, S i ups K, Kanoni S, Do R, Jun G, Hu Y, Kang HM, Xue C, Goel A, Fa all M, Duga S, Me lini PA, Assel a R, Gi elli D, Oli ie i O, Ma inelli N, Yin W, Reilly D, Spelio es E, Fox CS, H eem K, Holmen OL, Nikpay M, Fa low DN, Assimes TL, F anceschini N, Robinson J, No h KE, Ma in LW, DeP is o M, Gup a N, Esche SA, Jansson JH, Van Zuydam N, Palme CN, Wa eham N, Koch W, Mei inge T, Pe e s A, Lieb W, E bel R, Konig IR, K uppa J, Degenha d F, Go esman O, Bo inge EP, O’Donnell CJ, Psa y BM, Ballan yne CM, Abecasis G, O do as JM, Melande O, Wa kins H, O ho-Melande M, A dissino D, Loos RJ, McPhe son R, Wille CJ, E dmann J, Hall AS, Samani NJ, Deloukas P, Schunke H, Wilson JG, Koope be g C, Rich SS, T acy RP, Lin DY, Al shule D, Gab iel S, Nicke son DA, Ja ik GP, Cupples LA, Reine AP, Boe winkle E, Ka hi esan S. Loss-o - unc ion mu a ions in APOC3, iglyce ides, and co ona y disease. N Engl J Med 2014;371:22–31. 7 Yang J, Lee SH, Godda d ME, Vissche PM. GCTA: a ool o genome-wide complex ai analysis. Am J Hum Gene 2011;88:76–82. 8 Adzhubei IA, Schmid S, Peshkin L, Ramensky VE, Ge asimo a A, Bo k P, Kond asho AS, Sunyae SR. A me hod and se e o p edic ing damaging missense mu a ions. Na Me hods 2010;7:248–9. 9 Schwa z JM, Rödelspe ge C, Schuelke M, Seelow D. Mu a ionTas e e alua es disease-causing po en ial o sequence al e a ions. Na Me hods 2010;7:575–6. 10 Chun S, Fay JC. Iden ifica ion o dele e ious mu a ions wi hin h ee human genomes. Genome Res 2009;19:1553–61. 11 Romeo S, Pennacchio LA, Fu Y, Boe winkle E, Tybjae g-Hansen A, Hobbs HH, Cohen JC. Popula ion-based esequencing o ANGPTL4 unco e s a ia ions ha educe iglyce ides and inc ease HDL. Na Gene 2007;39:513–16. 12 an Leeuwen EM, Ka ssen LC, Deelen J, Isaacs A, Medina-Gomez C, Mba ek H, Kan e akis A, T ompe S, Pos mus I, Ve weij N, an Encke o DJ, Hu man JE, Whi e CC, Fei osa MF, Ba z TM, Manichaikul A, Joshi PK, Peloso GM, Deelen P, an Dijk F, Willemsen G, de Geus EJ, Milaneschi Y, Penninx BW, F ancioli LC, Menelaou A, Puli SL, Ri adenei a F, Ho man A, Oos a BA, F anco OH, Ma eo Leach I, Beekman M, , de C aen AJ, Uh HW, T oche H, Hocking LJ, Po eous DJ, Sa a N, Packa d CJ, Buckley BM, B ody JA, Bis JC, Ro e JI, Mychaleckyj JC, Campbell H, Duan Q, Lange LA, Wilson JF, Haywa d C, Polasek O, Vi a V, Rudan I, W igh AF, Rich SS, Psa y BM, Bo ecki IB, Kea ney PM, S o DJ, Ad ienne Cupples L; Genome o The Ne he lands Conso ium, Jukema JW, an de Ha s P, Sijb ands EJ, Ho enga JJ, Ui e linden AG, Swe z MA, an Ommen GJ, de Bakke PI, Eline Slagboom P, Boomsma DI, Wijmenga C, an Duijn CM. Genome o he Ne he lands popula ion-specific impu a ions iden i y an ABCA6 a ian associa ed wi h choles e ol le els. Na Commun 2015;6:6065. 13 Jones PM, Geo ge AM. The ABC anspo e s uc u e and mechanism: pe spec i es on ecen esea ch. Cell Mol Li e Sci 2004;61:682–99. 14 Towle HC. Glucose as a egula o o euka yo ic gene ansc ip ion. T ends Endoc inol Me ab 2005;16:489–94. 15 Den in R, Pégo ie JP, Benhamed F, Fou elle F, Fe é P, Fau eau V, Magnuson MA, Gi a d J, Pos ic C. Hepa ic glucokinase is equi ed o he syne gis ic ac ion o ChREBP and SREBP-1c on glycoly ic and lipogenic gene exp ession. J Biol Chem 2004;279:20314–26. 16 Den in R, Gi a d J, Pos ic C. Ca bohyd a e esponsi e elemen binding p o ein (ChREBP) and s e ol egula o y elemen binding p o ein-1c (SREBP-1c): wo key egula o s o glucose me abolism and lipid syn hesis in li e . Biochimie 2005;87:81–6. 17 Ma L, Robinson LN, Towle HC. ChREBP*Mlx is he p incipal media o o glucose-induced gene exp ession in he li e . J Biol Chem 2006;281:28721–30. 18 Uyeda K, Repa JJ. Ca bohyd a e esponse elemen binding p o ein, ChREBP, a ansc ip ion ac o coupling hepa ic glucose u iliza ion and lipid syn hesis. Cell Me ab 2006;4:107–10. 19 Alb ech sen A, G a up N, Li Y, Spa sø T, Tian G, Cao H, Jiang T, Kim SY, Ko neliussen T, Li Q, Nie C, Wu R, Sko e L, Mo is AP, Laden all C, Cauchi S, S ančáko á A, Ande sen G, As up A, Banasik K, Benne AJ, Bolund L, Cha pen ie G, Chen Y, Dekke JM, Doney AS, Do khan M, Fo sen T, F ayling TM, G o es CJ, Gui Y, Hallmans G, Ha e sley AT, He K, Hi man GA, Holmk is J, Huang S, Jiang H, Jin X, Jus esen JM, K is iansen K, Kuusis o J, Laje M, Lan ie i O, Li W, Liang H, Liao Q, Liu X, Ma T, Ma X, Manijak MP, Ma e M, Mok osiński J, Mo is AD, Mu B, Nielsen AA, Nijpels G, Nilsson P, Palme CN, Rayne NW, Rens öm F, Ribel-Madsen R, Robe son N, Rolandsson O, Rossing P, Schwa z TW; D.E.S.I.R. S udy G oup, Slagboom PE, S e ne M; DIAGRAM Conso ium, Tang M, Ta now L, Tuomi T, an’ Rie E, an Leeuwen N, Va ga TV, Ves ma MA, Walke M, Wang B, Wang Y, Wu H, Xi F, Yengo L, Yu C, Zhang X, Zhang J, Zhang Q, Zhang W, Zheng H, Zhou Y, Al shule D, ’ Ha LM, F anks PW, Balkau B, F oguel P, McCa hy MI, Laakso M, G oop L, Ch is ensen C, B andslund I, Lau i zen T, Wi e DR, Linnebe g A, Jø gensen T, Hansen T, Wang J, Nielsen R, Pede sen O. Exome sequencing-d i en disco e y o coding polymo phisms associa ed wi h common me abolic pheno ypes. Diabe ologia 2013;56:298–310. 20 Gu dasani D, Ca s ensen T, Tekola-Ayele F, Pagani L, Tachmazidou I, Ha ziko oulas K, Ka hikeyan S, Iles L, Polla d MO, Choudhu y A, Ri chie GR, Xue Y, Asimi J, Nsubuga RN, Young EH, Pomilla C, Ki inen K, Rocke K, Kamali A, Douma ey AP, Asiki G, Seeley J, Sisay-Joo F, Jallow M, Tollman S, Mekonnen E, Ekong R, Olji a T, B adman N, Bojang K, Ramsay M, Adeyemo A, Bekele E, Mo ala A, No is SA, Pi ie F, Kaleebu P, Kwia kowski D, Tyle -Smi h C, Ro imi C, Zeggini E, Sandhu MS. The A ican Genome Va ia ion P ojec shapes medical gene ics in A ica. Na u e 2015;517:327–32. an Leeuwen EM, e al.J Med Gene 2016;53:441–449. doi:10.1136/jmedgene -2015-103439 449 Genome-wide s udies g oup.bmj.com on No embe 11, 2016 - Published by h p://jmg.bmj.com/Downloaded om