Genetic analysis for a shared biological basis between migraine and coronary artery disease

ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. Gene ic analysis o a sha ed biological basis be ween mig aine and co ona y a e y disease Bendik S. Wins old, MD * Ch is ophe P. Nelson, PhD * Raine Malik, PhD Padh aig Go mley, PhD Ve ne i An ila, PhD Jason Vande Heiden, MSc Ka he ine S. Ellio , PhD Line M. Jacobsen, PhD P ii Pal a, PhD Naja Amin, PhD Boukje de V ies, PhD ABSTRACT Objec i e:T o a p p l y g en e i c a n a l y s i s o g e n o m e - w i d e a s s o c i a i o n d a a o s u d y h e e x e n a n d n a u e o a s h a e d b i o - logical basis be ween mig aine and co ona y a e y disease (CAD). Me hods: Fou sepa a e me hods o c oss-pheno ype gene ic analysis we e applied on da a om 2 la ge-scale genome-wide associa ion s udies o mig aine (19,981 cases, 56,667 con ols) and CAD (21,076 cases, 63,014 c on ols) . T he i s 2 me hod s quan i ie d he e x e n o o e lap ping is k a ian s and as s e ssed h e load o CAD isk loci in mig aineu s. Genomic egions o sha ed isk we e hen iden i ied by analysis o co a iance pa e ns be ween he 2 pheno ypes and by que ying known genome-wide signi ican loci. Resul s:We oun d a sig ni ic an o e lap o ge n e ic i s k loc i o m i g aine and CA D. Whe n s a i i e d by mig ain e sub - ype, his was limi ed o mig aine wi hou au a, and he o e lap was p o ec i e in ha pa ien s wi h mig aine had a lowe load o CAD isk alleles han con ols. Genes indica ed by 16 sha ed isk loci poin o mechanisms wi h po en ial oles in mig aine pa hogenesis and CAD, including endo helial dys unc ion (PHACTR1) and insulin homeos asis (GIP). Conclusions:The e sul s sugg es ha sha ed biolog ical p o cess es con ibu e o isk o mig aine and C AD, b u s u - p isingly his commonali y is es ic ed o mig aine wi hou au a and he impac is in opposi e di ec ions. Unde s and- ing he mechanisms unde lying hese p ocesses and hei opposi e ela ionship o mig aine and CAD may imp o e ou unde s anding o bo h diso de s. Neu ol Gene 2015;1:e10; doi: 10.1212/NXG.0000000000000010 Eija Hämäläinen, BS Tobias F eilinge , MD M. A an Ik am, MD Tho s en Kessle , MD GLOSSARY CAD 5 co ona y a e y disease; CARDIoGRAM 5 Co ona y AR e y DIsease Genome-Wide Replica ion And Me a-Analysis; CPSM 5 C oss-Pheno ype Spa ial Mapping; GWAS 5 genome-wide associa ion s udies; IHGC 5 In e na ional Headache Gene ics Conso ium; LD 5 linkage disequilib ium; MA 5 mig aine wi h au a; MO 5 mi g ai ne wi hou au a; S NP 5 single nucleo ide polymo phism. Ma kku Koi anen, MS Lannie Lig ha , PhD Geo ge McMahon, PhD Linda M. Pede sen, PhD Ch is ina Willenbo g, PhD Hong-Hee Won, PhD Jes Olesen, PhD Ville A o, MD Themis ocles L. Assimes, PhD S e an Blankenbe g, MD Do e I. Boomsma, PhD Lynn Che kas, PhD Geo ge Da ey Smi h, Dsc S ephen E. Eps ein, MD Jeane e E dmann, PhD Michel D. Fe a i, PhD Ha mu Göbel, PhD Alis ai S. Hall, PhD Ma jo-Rii a Ja elin, PhD Mikko Kallela, PhD Jaakko Kap io, PhD A u h o li s c on in ued o n nex pa g e Mig aine a ec s 19% o women and 11% o men wo ldwide and causes mo e yea s los o dis- abili y han any o he neu ologic diso de . 1,2 In abou one- hi d o pa ien s, headache a acks a e p eceded by ansien neu ologic symp oms e med mig aine au a, and mig aine wi h and wi hou au a (MA and MO, espec i ely) a e belie ed o ha e a pa ially dis inc pa hogenic basis. 3 I has long been assumed ha he ascula sys em is in ol ed in mig aine pa hogenesis, bu li le is known o he speci ic biological p ocesses in ol ed, and he ela i e impo ance o neu onal and ascula mechanisms emains con o e sial. 3 – 6 Suppo ing a ascula basis, epide- miologic s udies ha e ound an inc eased isk o s oke among pa ien s wi h mig aine, mos p onounced o MA. 7 Some ecen s udies indica e a simila isk inc ease o co ona y a e y disease (CAD), he mos common ascula diso de , al hough he associa ion is less ce ain han o s oke. 8 – 11 This aises he ques ion o whe he mig aine and ca dio ascula disease ha e a sha ed biological basis. Bo h mig aine and CAD ha e a s ong gene ic de e mina ion, and ecen genome-wide associa- ion s udies (GWAS) ha e iden i ied isk a ian s o each. I mig aine and CAD ha e a sha ed bio- logical basis, one migh an icipa e ha hey will also sha e gene ic a ian s ha a ec hei isk. In his s udy, we u ilized da a om 2 la ge-scale nono e lapping GWAS me a-analyses o mig aine ( he In e na ional Headache Gene ics Conso ium, IHGC) 12 and CAD (Co ona y AR e y DIsease Genome-Wide Replica ion And Me a-Analysis, CARDIoGRAM) 13 o quan i y sha ed gene ic isk. METHODS S udy coho s. Summa y s a is ics (p alue and e ec size) a single nucleo ide polymo phism (SNP) le el om 2 ecen ly pe o med me a-analyses o genome-wide associa ion da a on mig aine (IHGC)12 and CAD (CARDIoGRAM)13 we e used in *These au ho s con ibu ed equally o he manusc ip . Au ho a ilia ions a e p o ided a he end o he a icle. Funding in o ma ion and disclosu es a e p o ided a he end o he a icle. Go o Neu ology.o g/ng o ull disclosu e o ms. The A icle P ocessing Cha ge was paid by he au ho s. The CARDIoGRAM Conso ium and he In e na ional Headache Gene ics Conso ium coin es iga o s a e lis ed a Neu ology.o g/ng. This is an open access a icle dis ibu ed unde he e ms o he C ea i e Commons A ibu ion License 4.0 (CC BY), which pe mi s un es ic ed use, dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed. Neu ology.o g/ng © 2015 Ame ican Academy o Neu ology 1 ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. Seka Ka hi esan, MD Te ho Leh imäki, MD Ru h McPhe son, PhD Win ied Mä z, PhD Dale R. Nyhol , PhD Ch is ophe J. O ’ Donnell, MD Lydia Quaye, PhD Daniel J. Rade , MD Olli Rai aka i, PhD Robe Robe s, MD He ibe Schunke , MD Ma kus Schü ks, MD Alexand e F.R. S ewa , PhD Gisela M. Te wind , PhD Unnu Tho s einsdo i , PhD A n M.J.M. an den Maagdenbe g, PhD Co nelia an Duijn, PhD he p esen s udy. A e excluding o e lapping samples, he 2 s udies consis ed o 19,981 cases wi h mig aine s 56,667 con ols, and 21,076 cases wi h CAD s 63,014 con ols. A p opo ion o he mig aine cases we e pheno yped in su icien de ail o allow subclassi ica ion in o MO (6,413 cases, 32,745 con ols) and MA (4,940 cases, 37,557 con ols). In addi ion, indi idual-le el geno ype da a we e a ailable o a p opo ion o he mig aine coho s (6,350 mig aine cases s 15,069 con ols om he Ge man MA and MO coho s, Du ch LUMINA s udy, Finnish MA s udy, and he HUNT S udy, No way). All da a se s we e impu ed by using he HapMap elease 21 o 22 as e e ence. An o e iew o he s udy design and he included coho s is gi en in igu e 1. A de ailed desc ip ion o samples, geno yping, and associa ion analyses is gi en in e-Me hods, ables e-1 and e-2, and igu e e-1 a Neu ology.o g/ng. S anda d p o ocol app o als, egis a ions, and pa ien consen s. Fo all s udy coho s, pa icipa ion was based on in o med consen . Each s udy was app o ed by local esea ch e h- ics boa ds in he coun y whe e he s udy coho was collec ed. See o iginal publica ions o he 2 s udies o ull de ails o e hics and consen p ocedu es.12,13 Analy ic app oach. E alua ing ex en o o e lapping signals. To assess whe he mo e associa ion signals we e sha ed be ween p uned SNP se s in o de o op imize sensi i i y. Using each se o CAD isk SNPs, we calcula ed a pe -indi idual CAD polygenic isk sco e by summing he numbe o CAD isk alleles (o expec ed allele coun s o impu ed SNPs), each weigh ed by he log odds a io om he CAD s udy. We subsequen ly assessed whe he CAD polygenic isk sco e was associa ed wi h mig aine s a us by applying a logis ic eg ession model o he e ec o CAD polygenic isk sco e (con inuous) on mig aine s a us (case, con- ol), adjus ed o sex and dummy-coded co a ia es ep esen ing he 6 indi idual mig aine s udy coho s. Iden i ying sha ed isk loci. In o de o iden i y sha ed isk loci be ween mig aine and CAD, we applied a no el me hod, C oss-Pheno ype Spa ial Mapping (CPSM; see e-Me hods o an o e iew). This me hod compa es 2 se s o p alues om GWAS in o de o ind g oups o SNPs a which hey a e co ela ed and hus iden i y sha ed pa e ns o associa ion. We applied his me hod o he 2,342,101 o e lapping SNPs om he mig aine and CAD s udies and selec ed genomic egions wi h signal abo e he 99.95 h pe cen ile o 1,000 pe mu a ions o u he analysis. Po en ial e - ec s o he sha ed associa ion loci on egional gene exp ession (cis e ec ) we e examined using an exis ing exp ession quan i a i e ai locus da abase om pe iphe al blood17 (e-Me hods). Las ly, we analyzed loci wi h p e iously epo ed genome- wide signi ican associa ion o mig aine o CAD (summa ized 12,13 Maija Wessman, PhD he mig aine and CAD s udies han would be expec ed by chance, in he o iginal publica ions). The lead SNP a each locus Tobias Ku h, MD Ch is ian Kubisch, MD Ma in Dichgans, MD Daniel I. Chasman, PhD Ch is Co sapas, PhD John-Anke Zwa , MD Nilesh J. Samani, FRCP Aa no Palo ie, MD Fo he CARDIoGRAM Conso ium and he we used a se o 2,342,101 o e lapping SNPs ha we e di ec ly yped o impu ed in bo h s udies. Following he same p ocedu e as desc ibed in a p e ious s udy, 14 we i s so ed he SNPs by associa ion p alue o mig aine. S a ing om he op o he lis , all subsequen SNPs wi h linkage disequilib ium (LD) 2 . 0.05 (based on HapMap CEU elease 27) we e emo ed. This p ocess was epea ed un il a se o 92,654 SNPs in app oxima e linkage equilib ium emained. Fo each o 5 sepa a e p alue cu o s (1 3 10 2 2 ,1 3 10 2 3 ,1 3 10 2 4 ,1 3 10 2 5 , and 1 3 10 2 6 ), we coun ed he numbe o SNPs abo e and below he cu o in each o he 2 s udies, esul ing in one 2 3 2 able o each p alue cu o . The was c oss-analyzed o associa ion o he o he pheno ype, Bon- e oni co ec ing o he numbe o SNPs es ed. All 13 o 13 epo ed mig aine loci and 22 o 25 epo ed CAD loci we e a ailable in ou da a se and could be es ed (excluding CAD isk SNPs s17465637, s1746048, and s12413409). RESULTS Compa ing nominally signi ican SNPs om he mig aine and CAD GWAS, we ound an o e lap o associa ion signals in excess o wha would be expec ed by chance ( able 1). An o e lap o signals was seen o SNPs wi h p alues #1 3 1022, Fishe exac es was used o es ima e de ia ion om he expec ed 2 4 2 6 In e na ional Headache dis ibu ion, and alse disco e y a e co ec ion was pe o med on 1 3 10 , and 1 3 10 . This was suppo ed by Gene ics Conso ium Co espondence o D . Palo ie: aa no@b oadins i u e.o g See edi o ial Supplemen al da a a Neu ology.o g/ng all 6 es s using he p.adjus unc ion in R.15 A co ec ed Fishe p , 0.01 was aken o indica e an excess o o e lapping signals. In o de o ob ain a mo e obus es ima e o he signi icance o he obse ed o e lap, his was also assessed h ough pe mu a ions. In each pe mu a ion cycle, he ela ion o p alues o SNPs was andomized wi hin each o he LD-p uned mig aine and CAD da a se s, and a Fishe p o o e lap was calcula ed o each p alue cu o . We gene a ed 100,000 pe mu a ions o p oduce an empi ical null dis ibu ion o p alues. In an equi alen manne , seconda y analyses we e pe o med o MO (83,373 o e lapping SNPs a e LD p uning) and MA (88,031 o e lapping SNPs a e LD p uning). Polygenic isk sco e analysis. I sha ed gene ic isk a ian s a e in pa o ully esponsible o como bidi y be ween mig aine and CAD, we would expec an accumula ion o CAD isk alleles in mig aineu s. To es his hypo hesis, we used he 6 mig aine coho s in which indi idual-le el geno ype da a we e a ailable o analysis (6,350 mig aineu s s 15,069 con ols; igu e 1). Fo each mig aine case o con ol, we calcula ed a CAD polygenic isk sco e based on a p e iously published me hod.16 We i s gen- e a ed 3 se s o CAD isk SNPs by selec ing SNPs wi h s ong (p , 5 3 1028; 149 SNPs), mode a e (p , 1 3 1024; 1,631 SNPs), o weak ( p , 1 3 10 2 2 ; 36,384 SNPs) associa ion o CAD among he 2,342,101 SNPs wi h in o ma ion in bo h mig aine and CAD s udies. As sugges ed in he o iginal desc ip ion o he me hod,16 he analysis was based on non–LD- pe mu a ion es ing, which indica ed a sha ing o associa ion signals a p alue cu o 1 3 10 2 5 as well. Fo e e ence, he ull lis o SNPs wi h associa ion p alue # 1 3 10 2 2 o bo h CAD and mig aine is gi en in able e-3. Seconda y analyses by mig aine sub ype e ealed an o e lap o associa ion signals be ween MO and CAD a all p alue cu o s (1 3 10 2 2 ,1 3 10 2 3 ,1 3 10 2 4 ,1 3 10 2 5 , and 1 3 10 2 6 ), while no o e lap was seen be ween MA and CAD a any o he p alue cu o s. The di ec ion o e ec o o e - lapping associa ion signals did no consis en ly ag ee be ween mig aine and CAD, as e idenced by nonsigni - ican binomial p alues o conco dance ( able 1). To examine his u he , he second analysis com- pa ed he load o gene ic isk a ian s o CAD be ween mig aineu s and con ols, using indi idual- le el da a. The esul s indica ed ha a high CAD pol- ygenic isk sco e was associa ed wi h a educed isk o mig aine ( igu e 2, u he de ails in able e-4). Fo mig aine o e all, his associa ion was seen o only he mode a e CAD isk SNP se ( p 5 0.007). Seconda y analyses e ealed a simila , bu mo e p onounced, associa ion be ween CAD polygenic isk sco e and 2 Neu ology: Gene ics ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. CPSM 5C oss-Pheno ype Spa ial Mapping. MO ( p 5 1.5 3 10 2 4 and 5.1 3 10 2 4 o he mod- e a e and s ong CAD isk SNP se s, espec i ely). No associa ion was seen o MA. In he analysis o he weak CAD isk SNP se , he e was no associa ion o CAD gene ic isk sco e o ei he mig aine ca ego y, indica ing ha he obse ed associa ions we e d i en by a ai ly limi ed numbe o loci ha a e a leas mode a ely associa ed wi h CAD. These indings we e consis en ac oss men and women ( igu e e-2) and ac oss indi idual independen coho s wi hin he same mig aine sub ype ( igu e e-3). CPSM yielded 16 loci ha o e lapped be ween mig aine and CAD ( able 2; igu e e-4). De ails o he mos signi ican mig aine and CAD SNPs a each locus a e gi en in able e-5. The s onges e i- dence o sha ed associa ion was seen a 6p24 (locus no. 1 o able 2), whe e bo h CAD and mig aine showed genome-wide signi ican signals wi hin he PHACTR1 gene (CAD: s4714955, p 5 9.8 3 10 2 11 ; mig aine: s9349379, p 5 5.9 3 10 2 9 ). The second s onges o e lapping signal was on 17q21 (locus no. 2), whe e he lead CAD SNP ( s46522, p 5 2.6 3 10 2 7 ) was in agenic in UBE2Z , whe eas he lead mig aine SNP ( s11079844, p 5 3.1 3 10 2 5 ) was in e genic be ween SNF8 and GIP . I is in e es ing ha bo h lead SNPs a e in high LD ( 2 . 0.9) wi h 2 unc ional a ian s in GIP : Se 103Gly ( s2291725) and a splice si e a ian ( s2291726) ha is p edic ed o lead o a p ema u ely unca ed an- sc ip 18 ( able e-6). The locus was also ound o ha e a po en ial e ec on he exp ession le el o UBE2Z ( able e-7). Lead SNPs in 5 loci we e in high LD ( 2 . 0.8) wi h nonsynonymous o splice si e a ian s in nea by genes ( able e-6). Ten o he 16 loci showed opposi e di ec ion o e ec o mig aine and CAD. In he seconda y analyses, 12 o he 16 lead mig aine SNPs had a lowe associa ion p alue in MO han in MA (2- ailed binomial p 5 0.08), and all 16 SNPs Neu ology: Gene ics 3 Figu e 1 S udy design and included coho s ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. Table 1 Analysis o he ex en o o e lapping signals be ween mig aine and CAD pValue o o e lap pValue o o e lap Conco dance o Binomial p alue Signal de ini ion (p alue cu o ) O e lapping SNPs (Fishe exac es ) a (pe mu a ion es ) a o e lapping SNPs b o conco dance All mig aine ( o al no. SNPs: 92,654) 1E-2 146 1.1E-05 c 6.0E-05 c 0.534 0.18 1E-3 7 0.099 0.056 0.571 0.23 1E-4 2 9.3E-03 c 7.9E-03 c 0 0.75 1E-5 1 0.014 2.1E-04 c 0 0.50 1E-6 1 5.2E-03 c 3.5E-03 c 0 0.50 Mig aine wi hou au a ( o al no. SNPs: 83,373) 1E-2 113 1.6E-04 c 1.5E-05 c 0.510 0.87 1E-3 8 1.3E-03 c 1.1E-04 c 0.442 0.86 1E-4 3 2.0E-04 c 1.5E-05 c 0.250 0.88 1E-5 1 8.5E-03 c 1.5E-05 c 0 0.50 1E-6 1 3.0E-03 c 1.5E-05 c 0 0.50 Mig aine wi h au a ( o al no. SNPs: 88,031) 1E-2 107 0.13 0.11 0.523 0.28 1E-3 1 1.0 0.64 0.523 0.50 1E-4 0 1.0 1.0 NA NA 1E-5 0 1.0 1.0 NA NA 1E-6 0 1.0 1.0 NA NA Abb e ia ions: CAD 5co ona y a e y disease; NA 5no applicable; SNP 5single nucleo ide polymo phism. aFalse disco e y a e co ec ed p alues. bP opo ion o o e lapping associa ion signals ha ing he same di ec ion o e ec in mig aine and CAD. c p , 0.01. Re sul s a e gi e n as od d s a i os wi h 9 5 % co n i d e nc e i n e al s . S ep a a e l i nes a e s ho wn o al l mig aine (blue), mig aine wi hou au a (g een), and mig aine wi h au a ( ed). The co ona y a e y disease (CAD) polygenic isk sco e was calcula ed based on single nucleo ide polymo phisms (SNPs) wi h weak (p , 1 3 1022), mode a e (p , 1 3 1024), o s ong (p , 5 3 1028) associa ion o CAD in he Co ona y AR e y DIsease Genome-Wide Replica ion And Me a-Analysis s udy. 4 Neu ology: Gene ics had he same e ec di ec ion in each o he 2 mig aine sub ypes. Local Manha an plo s and co a iance plo s o he iden i ied loci a e gi en in igu e e-4. When conside ing p e iously epo ed isk loci o mig aine and CAD, 3 CAD isk SNPs we e associa ed o mig aine a s udy-wide signi icance, and 2 mig aine isk SNPs we e associa ed o CAD ( able 3). These co espond o loci no. 1, 2, 3, 11, and 14 as iden i ied by he CPSM me hod and co obo a e he e idence o sha ed gene ic isk a hese loci. DISCUSSION In his s udy, we used da a om 2 ecen ly pe o med la ge-scale nono e lapping GWAS o examine sha ed gene ic isk be ween mig aine and CAD. We ound ha associa ion signals o e lapped in excess o wha would be expec ed by chance. S a i ying by mig aine sub ype u he e ealed ha MO and MA beha ed di e en ly. MO had a gene ic o e lap wi h CAD, whe eas MA did no . These esul s a e unexpec ed, gi en he epidemiologic e idence ha como bidi y wi h CAD is mo e common in MA han MO. Pa ien s wi h MA we e ound o ha e a 2- old inc eased isk o CAD 8,10 and an inc eased isk o CAD- ela ed mo ali y, 9,11 al hough one c oss- sec ional s udy ailed o ind an associa ion be ween Figu e 2 Associa ion be ween co ona y a e y disease polygenic isk sco e and he p esence o mig aine ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. Table 2 O e lapping associa ion loci be ween mig aine and CAD as iden i ied by CPSM analysis Posi ion (Mb)b Locus Locus no. a Ch band Le ma gin Righ ma gin Size (kb) Peak SNP Peak heigh Genes wi hin locus c 16p24 12.929 13.187 257.386 s7454157 25.2 PHACTR1 217q21 44.282 44.512 230.813 s12601858 13.2 CALCOCO2,ATP5G1,UBE2Z,SNF8,GIP,IGF2BP1 36q16 96.917 97.188 271.548 s12529248 7.8 UFL1,FHL5 412q24 110.241 111.064 823.633 s7962138 7.4 CUX2,FAM109A,SH2B3,ATXN2,BRAP,ACAD10, ALDH2 , MAPKAPK5 - AS1 , MAPKAPK5 , ADAM1A , TMEM116 , ERP29 , NAA25 , TRAFD1 517p11 17.603 17.992 389.086 s9890341 6.9 RAI1,SMCR5,SREBF1,MIR33B,TOM1L2,LRRC48, ATPAF2 , GID4 , DRG2 , MYO15A 616q23 73.827 74.094 266.738 s4888396 5.2 BCAR1,CFDP1,TMEM170A,CHST6 710q24 104.823 105.178 354.585 s7067970 5.1 CNNM2,NT5C2,LOC729020,INA,PCGF6,TAF5, USMG5 , MIR1307 , PDCD11 82q33 203.375 203.52 145.53 s6435169 4.8 ICA1L,WDR12,ALS2CR8 910q24 104.569 104.785 216.416 s1538204 4.1 CYP17A1,C10o 32,C10o 32-AS3MT,AS3MT, CNNM2 10 6q13 72.182 72.403 220.605 s12207845 3.1 LINC00472 11 8q21 89.53 89.69 160.492 s1352317 3.1 None 12 19q13 46.543 46.664 120.921 s3810174 2.9 TGFB1,B9D2,TMEM91,EXOSC5,BCKDHA,B3GNT8, ATP5SL , C19o 69 , LOC100505495 13 12q24 109.501 109.714 213.015 s16940933 2.8 PPTC7,TCTN1,HVCN1,PPP1CC 14 8q21 89.382 89.525 143.754 s6984041 2.7 MMP16 15 16q24 88.099 88.191 91.257 s17775174 2.6 SPG7,RPL13,SNORD68,CPNE7 16 9p21 23.429 23.505 75.25 s10811931 2.5 None Abb e ia ions: Ch 5ch omosome; CAD5co on a y a e y di sea se; CPSM 5C oss-Pheno ype Spa ial Mapping; peak heigh 5 alue o co a iance signal a a p ex o h e p e a k ; S NP 5single nucleo ide polymo phism. aSo ed by dec easing peak heigh . bPosi ions e e o build NCBI36/hg18. cRe Seq genes. CAD and any mig aine sub ype.19 S udies no di e en ia ing on mig aine sub ype ha e been less conclusi e, wi h some8,9,20,21 bu no o he s19,22 indica ing an inc eased isk o CAD ela ed o mig aine o e all. Fo MO, we ound a clea o e lap o associa ion signals wi h CAD, whiche e p alue cu o was used o de ine signals. In iguingly, he impac was in he opposi e di ec ion, in ha pa ien s wi h MO had a lowe load o CAD isk alleles han mig aine- ee con ols. This associa ion seemed o be d i en by a limi ed numbe o loci. Only a p opo ion o he included mig aine pa ien s we e pheno yped in su i- cien de ail o allow subclassi ica ion in o MA o MO. When using he conside ably la ge se o all mig aine pa ien s, a simila associa ion was seen as o MO, likely d i en by his mig aine sub ype. While he esul s sugges ha he e a e sha ed common isk a ian s be ween mig aine and CAD, hey do no indica e ha hese a ian s explain como bidi y be ween he 2 diso de s. The opposi e di ec ion o e ec o some o he loci is consis en wi h a ecen GWAS in which he mig aine and CAD isk SNP s9349379 (in PHACTR1) was associa ed wi h ce ical a e y dissec- ion, wi h e ec in he same di ec ion as o mig aine bu opposi e o CAD. 23 Two u he mig aine SNPs showed e idence o associa ion o ce ical a e y dis- sec ion wi h he same e ec di ec ion as o mig aine ( s11172113 in LRP1 and s13208321 in FHL5 , he la e iden i ied as locus 3 in he cu en s udy) bu opposi e di ec ion o CAD. The signi ican sha ing o isk loci be ween mig aine and CAD may e lec ha hey in ol e some o he same biological p ocesses. Expe imen al s udies will be needed o cla i y his and whe he he sha ed isk loci can gi e in o ma ion on ascula mechanisms in ol ed in mig aine pa hogenesis. The lack o o e lapping associa ion signals be ween MA and CAD may indica e ha he 2 diso - de s ha e sepa a e and non ela ed gene ic back- g ounds. Howe e , i may also esul om insu icien powe o de ec sha ed common gene ic isk ac o s o his mig aine sub ype. This is consis- en wi h he ela i e ailu e so a in iden i ying com- mon isk a ian s o MA; despi e a leas as high he i abili y and compa able s udy sample sizes, only one genome-wide signi ican locus has been iden i ied Neu ology: Gene ics 5 ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. o MA, compa ed o 9 o MO.12,24–27 I is possible ha MA is a mo e he e ogeneous diso de o is in lu- enced by a e and low- equency a ian s no cap- u ed by cu en impu a ion panels.12,28 La ge s udies ha also in e oga e a e a ian s will be needed o de e mine he gene ic basis o MA and i s po en ial o e lap wi h ca dio ascula disease. Six o he o e lapping loci ha e p e iously been associa ed wi h CAD a genome-wide signi ican le - els (loci 1, 2, 4, 7, 8, and 9 o able 2),13,29 and 2 wi h mig aine (loci 1 and 3).12,25 The s onges o e lapping egion (locus 1) is en i ely in agenic in PHACTR1 (which encodes phospha ase and ac in egula o 1 p o- ein). This locus is associa ed wi h bo h mig aine and CAD a genome-wide signi ican le els in he cu en and p e ious s udies13,25,30 and has also been associ- a ed wi h co ona y a e y calci ica ion and s oke.31,32 PHACTR1 is highly exp essed in he b ain, and i s ansc ip is an impo an egula o o synap ic ac i - i y and dend i ic mo phology h ough he con ol o p o ein phospha ase 1 and ac in binding.33 Mo e ecen ly, PHACTR1 has been iden i ied as a key eg- ula o o endo helial unc ion, including endo helial cell su i al and angiogenesis,34 and i is associa ed wi h al e ed asomo o one.35 Bo h endo helial and asomo o dys unc ions ha e been implica ed in mig aine,36,37 and his locus o e s a po en ial ocus o u u e s udies. Al e na i ely, he pleio opic e ec s o his gene on bo h synap ic and ascula unc ions may gi e ise o independen causal pa hways o he 2 diso de s. The second s onges o e lapping egion (locus 2) is a p e iously iden i ied isk locus o CAD. 13 The lead CAD ( s46522) and mig aine a ian s ( s11079844) a e in s ong LD ( 2 5 0.94), and bo h a e in s ong LD ( 2 . 0.90) wi h 2 po en ially unc- ional a ian s in GIP (which encodes gas ic inhibi- o y polypep ide). GIP egula es glucose-induced insulin elease om panc ea ic b -cells and helps e- sensi ize he insulin esponse. 38 I is also exp essed in he b ain, whe e i may be in ol ed in p oli e a ion o neu onal p ogeni o cells. 39 Whe he GIP is in ol ed in he obse ed endency o insulin esis ance and me abolic synd ome in mig aineu s should be in es iga ed. 40 S eng hs o his s udy include he use o la ge- scale nono e lapping GWAS o mig aine and CAD, s ingen quali y con ol measu es, and su icien ly ich pheno yping o allow seconda y analyses o he 2 mig aine sub ypes. Ne e heless, some limi a ions should also be acknowledged. Fi s , only summa y s a is ics and no indi idual-le el geno ype da a we e a ailable o he majo i y o he samples included in his s udy. Second, in each included coho , pheno- ype in o ma ion was a ailable on only mig aine o CAD, no bo h. This p e en ed us om pe o ming 6 Neu ology: Gene ics Abb e ia ions: Ch 5 ch omosome; CAD 5 co ona y a e y disease; SNP 5 single nucleo ide polymo phism . Only SNPs wi h signi ican associa ion o he o he pheno ype a e Bon e oni co ec ion a e shown ( p , 0.0038 o p e iously epo ed m ig aine l oci in he CAD sample, and p , 0.0023 o p e iously epo ed CAD loci in he m ig aine s am ple). p alues may di e om hose epo ed in he o iginal s udies since o e lapping samples we e excluded in he cu en s udy. a Di ec ion o e ec : SNPs wi h he same e ec di ec ion o associa ion o CAD and mig aine a e ma ked as plus ( 1 ) , opposing e ec di ec ion i s ma ked as minus ( 2 ) . b Signi ican c oss-pheno ype p alue. Table 3 C oss-analysis o loci p e iously epo ed o show genome-wide signi ican associa ion wi h mig aine o CAD CAD Mig aine Mig aine wi hou au a Mig aine wi h au a Lead SNP Ch band Repo ed gene(s) pValue Odds a io(SE) pValue Odds a io(SE) Di .a pValue Odds a io(SE) Di .a pValue Odds a io(SE) Di . a P e iously epo ed loci o mig aine in he CAD sample s9349379 6p24 PHACTR1 8.97E-08 b 1.1 (0.019) 5.88E-09 1.09 (0.015) 2 2.52E-10 1.17 (0.029) 2 0.236 1.03(0.026) 2 s10504861 8q21 MMP16 1.96E-03 b 1.06 (0.02) 1.71E-03 1.05 (0.017) 1 3.36E-08 1.17 (0.032) 1 0.329 1.03(0.031) 1 s13208321 6q16 FHL5 2.53E-03 b 1.05 (0.018) 1.41E-10 1.1 (0.016) 2 1.35E-12 1.19 (0.029) 2 7.40E-04 1.1(0.03) 2 P e iously epo ed loci o CAD in he mig aine sample s12526453 6p24 PHACTR1 9.72E-09 1.09 (0.017) 8.18E-06 b 0.94 (0.013) 2 7.80E-08 b 0.88 (0.02) 2 0.399 0.98(0.025) 2 s46522 17q21 UBE2Z,GIP, 2.58E-07 0.93 (0.013) 1.24E-04 b 1.05 (0.013) 2 6.93E-03 1.06 (0.022) 2 0.081 1.04(0.024) 2 ATP5G1,SNF8 ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. mo e in-dep h analyses, including analysis o po en- ial gene-gene in e ac ions o iden i ica ion o CAD isk loci speci ic o mig aineu s. Thi d, conside able e o was de o ed o he ca e ul a oidance o sha ed con ols be ween s udies, and s ingen quali y con- ol measu es wi hin each da a se we e en o ced o educe he isk o spu ious e ec s esul ing om biases wi hin he da a se s. Ne e heless, we canno ule ou sub le biases ha could a ec he cu en e- sul s. Two such conce ns a e he e ec s o mig aine on su i al and he possibili y ha mig aineu s may be mo e likely o seek medical ea men and he e- o e be unde close su eillance wi h ega ds o o he diso de s. Fu u e e o s should aim o eplica e hese indings in su icien ly la ge p ospec i e da a se s whe e bo h pheno ypes a e measu ed in he same indi iduals. Ou s udy p o ides no el insigh s in o he ela- ionship be ween mig aine and CAD. In iguingly, and unexpec edly, he e was no gene ic o e lap be ween MA and CAD, o which epidemiologic s udies sugges como bidi y, bu he e was compel- ling e idence o a gene ic o e lap be ween MO and CAD, whe e he impac o isk a ian s o e all was in opposi e di ec ion o he 2 diso de s. The e- sul s do no demons a e ha sha ed common gene ic isk ac o s d i e como bidi y be ween he 2 diso - de s. Howe e , dissec ing he mechanisms unde lying he sha ed isk loci may imp o e ou unde s anding o bo h diso de s. AUTHOR AFFILIATIONS F om he Depa men o Neu ology (B.S.W., J.-A.Z.) and FORMI (B.S.W., L.M.J., L.M.P., J.-A.Z.), Oslo Uni e si y Hospi al, Oslo, No way; Ins i u e o Clinical Medicine (B.S.W., J.-A.Z.), Uni e si y o Oslo, No way; Well- come T us Sange Ins i u e (B.S.W., P.G., V. An ila, P.P., E.H., A.P.), Wellcome T us Genome Campus, Camb idge, Uni ed Kingdom; Depa - men o Ca dio ascula Sciences (C.P.N., N.J.S.), Uni e si y o Leices e , Clinical Sciences Wing and Na ional Ins i u e o Heal h Resea ch Leices e Biomedical Resea ch Uni in Ca dio ascula Disease (C.P.N., N.J.S), Glen- ield Hospi al, Leices e , Uni ed Kingdom; Ins i u e o S oke and Demen ia Resea ch (R. Malik, T.F., M.D.), Klinikum de Uni e si a Munchen, Ludwig-Maximilians-Uni e si a , Munich, Ge many; Munich Clus e o Sys ems Neu ology (SyNe gy) (R. Malik, M.D.), Munich, Ge many; P o- g am in Medical and Popula ion Gene ics (P.G., V. An ila, H.-H.W., S.K., C.C., A.P.) and S anley Cen e o Psychia ic Resea ch (V. An ila, A.P.), B oad Ins i u e, Camb idge, MA; Psychia ic & Neu ode elopmen al Gene - ics Uni (P.G., A.P.), Depa men o Psychia y, Analy ic and T ansla ional Gene ics Uni (V. An ila, A.P.), Depa men o Medicine, Cen e o Human Gene ic Resea ch (H.-H.W., S.K.), Ca dio ascula Resea ch Cen e (H.-H.W., S.K.), and Depa men o Neu ology (A.P.), Massachuse s Gen- e al Hospi al, Bos on, MA; Di ision o P e en i e Medicine (T. Ku h, D.I.C.), B igham and Women’s Hospi al and Depa men o Medicine (V. An ila, H.-H.W., S.K.), Ha a d Medical School, Bos on, MA; Depa - men o Gene ics (J.V.H., C.C.) and Depa men o Neu ology (C.C.), Yale Uni e si y School o Medicine, New Ha en, CT; Wellcome T us Cen e o Human Gene ics (K.S.E.), Uni e si y o Ox o d, Uni ed Kingdom; Depa men o Public Heal h (J.K.), Hjel Ins i u e and Ins i u e o Molec- ula Medicine Finland (P.P., E.H., J.K., M.W., A.P.), Uni e si y o Helsinki, Finland; Depa men o Epidemiology (N.A., M.A.I., C. .D.), Depa men o Radiology (M.A.I.), and Depa men o Neu ology (M.A.I.), E asmus Uni e si y Medical Cen e, Ro e dam, he Ne he lands; Depa men o Human Gene ics (B.d.V., A.M.J.M. .d.M.) and Depa men o Neu ology (M.D.F., G.M.T., A.M.J.M. .d.M.), Leiden Uni e si y Medical Cen e, Leiden, he Ne he lands; Depa men o Neu ology and Epilep ology and He ie-Ins i u e o Clinical B ain Resea ch (T.F.), Uni e si y o Tübingen, Ge many; Deu sches He zzen um München (T. Kessle , H.S.), Technische Uni e si ä München, Munich, Ge many; DZHK (Ge man Cen e o Ca - dio ascula Resea ch) (T. Kessle , H.S.), Pa ne Si e Munich Hea Alliance, Munich, Ge many; Ins i u e o Heal h Sciences (M. Koi anen, M.-R.J.) and Biocen e Oulu (M.-R.J.), Uni e si y o Oulu, Finland; Depa men o Bio- logical Psychology (L.L., D.I.B.), VU Uni e si y and EMGO1Ins i u e o Heal h and Ca e Resea ch (L.L.), VU Uni e si y Medical Cen e, Ams e - dam, he Ne he lands; Medical Resea ch Council (MRC) In eg a i e Epide- miology Uni a he Uni e si y o B is ol (G.M., G.D.S.), Uni ed Kingdom; Ins i u ü In eg a i e und Expe imen elle Genomik (C.W., J.E.), Uni e si- ä zu Lübeck, Lübeck, Ge many; DZHK (Ge man Resea ch Cen e o Ca dio ascula Resea ch) (C.W., J.E.), Pa ne Si e Hambu g/Lübeck/Kiel, Lübeck, Ge many; Depa men o Neu ology (J.O.), Glos up Hospi al, Uni e si y o Copenhagen, Denma k; Depa men o Neu ology (V. A o, M. Kallela), Helsinki Uni e si y Cen al Hospi al, Helsinki, Finland; Depa - men o Medicine (T.L.A.), S an o d Uni e si y School o Medicine, S an o d, CA; Medizinische Klinik und Poliklinik (S.B.), Uni e si ä smedizin Mainz, Johannes-Gu enbe g Uni e si ä Mainz, Ge many; Depa men o Twin Resea ch and Gene ic Epidemiology (L.C., L.Q.), King’s College London, Uni ed Kingdom; MedS a Hea Ins i u e (S.E.E.), Washing on Hospi al Cen e , Washing on, DC; Kiel Pain and Headache Cen e (H.G.), Kiel, Ge many; Di ision o Ca dio ascula and Neu onal Remodelling (A.S. H.), Mul idisciplina y Ca dio ascula Resea ch Cen e, Leeds Ins i u e o Gene ics, Heal h and The apeu ics, Uni e si y o Leeds, Uni ed Kingdom; Depa men o Child en, Young People and Families (M.-R.J.) and Depa - men o Men al Heal h and Subs ance Abuse Se ices (J.K.), Na ional Ins i- u e o Heal h and Wel a e, Helsinki, Finland; Depa men o Epidemiology and Bios a is ics (M.-R.J.), School o Public Heal h, MRC– Heal h P o ec ion Agency (HPA) Cen e o En i onmen and Heal h, Fac- ul y o Medicine, Impe ial College, London, Uni ed Kingdom; Uni o P ima y Ca e (M.-R.J.), Oulu Uni e si y Hospi al, Oulu, Finland; Depa - men o Clinical Chemis y (T.L.), Fimlab Labo a o ies, Tampe e, Finland; Uni e si y o Tampe e School o Medicine (T.L.), Finland; The John and Jenni e Ruddy Canadian Ca dio ascula Gene ics Cen e (R. McPhe son, R.R., A.F.R.S.) and A he ogenomics Labo a o y (R. McPhe son), Uni e si y o O awa Hea Ins i u e, On a io, Canada; Synlab Cen e o Labo a o y Diagnos ics Heidelbe g (W.M.), Heidelbe g, Ge many; Clinical Ins i u e o Medical and Chemical Labo a o y Diagnos ics (W.M.), Medical Uni e si y o G az, Aus ia; Ins i u e o Public Heal h (W.M.), Social and P e en i e Medicine, Medical Facul y Manneim, Uni e si y o Heidelbe g, Ge many; Ins i u e o Heal h and Biomedical Inno a ion (D.R.N.), Queensland Uni e si y o Technology, B isbane, Aus alia; Na ional Hea , Lung, and Blood Ins i u e’s F amingham Hea S udy (C.J.O.), F amingham, MA; Depa men o Medicine, Ins i u e o T ansla ional Medicine and The apeu ics, and Ca dio ascula Ins i u e (D.J.R.), Uni e si y o Pennsyl ania, Philadelphia, PA; Depa men o Clinical Physiology and Nuclea Medicine (O.R.), Tu ku Uni e si y Hospi al, Tu ku, Finland; Resea ch Cen e o Applied and P e en i e Ca dio ascula Medicine (O.R.), Uni e si y o Tu ku, Finland; Depa men o Neu ology (M.S.), Uni e si y Hospi al Essen, Essen, Ge many; deCODE gene ics (U.T.), Reykja ik, Iceland; Facul y o Medicine (U.T.), Uni e si y o Iceland, Reykja ik, Iceland; Ins i u e o Gene ics (M.W.), Folkhalsan Resea ch Cen e , Helsinki, Finland; Ins i u Na ional de la San ee de la Reche che Medicale (INSERM) Resea ch Cen e o Epidemiology and Bios a is ics (U897) Team–Neu oepidemiology (T. Ku h), Bo deaux, F ance; Uni e si y o Bo deaux (T. Ku h), F ance; and Ins i u e o Human Gene ics (C.K.), Uni- e si y Medical Cen e Hambu g-Eppendo , Hambu g, Ge many. AUTHOR CONTRIBUTIONS Bendik S. Wins old and Ch is ophe P. Nelson: pe o med s a is ical analysis; concei ed and designed he s udy; analyzed and in e p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con en . Raine Malik: concei ed and designed he s udy; analyzed and in e p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con en . Padh aig Go mley, Ve ne i An ila, Jason Vande Heiden, Ka he ine S. Ellio , Line M. Jacobsen, and P ii Pal a: analyzed and in e p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con en . Naja Neu ology: Gene ics 7 ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. Amin, Boukje de V ies, Eija Hämäläinen, Tobias F eilinge , M. A an Ik am, Tho s en Kessle , Ma kku Koi anen, Lannie Lig ha , Geo ge McMahon, Linda M. Pede sen, Ch is ina Willenbo g, and Hong-Hee Won: con ibu ed da a; d a ed/ e ised he manusc ip o con en . Jes Olesen: analyzed and in e p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con en . Ville A o, Themis ocles L. Assimes, S e an Blankenbe g, Do e I. Boomsma, Lynn Che kas, Geo ge Da ey Smi h, S ephen E. Eps ein, Jeane e E dmann, Michel D. Fe a i, Ha mu Göbel, Alis ai S. Hall, Ma jo-Rii a Ja elin, Mikko Kallela, Jaakko Kap io, Seka Ka hi esan, Te ho Leh imäki, Ru h McPhe son, Win ied Mä z, Dale R. Nyhol , Ch is ophe J. O’Donnell, Lydia Quaye, Daniel J. Rade , Olli Rai aka i, Robe Robe s, He ibe Schunke , Ma kus Schü ks, Alexand e F.R. S ewa , Gisela M. Te wind , Unnu Tho s einsdo i , A n M.J.M. an den Maagdenbe g, Co nelia an Duijn, and Maija Wessman: con ibu ed da a; d a ed/ e ised he man- usc ip o con en . Tobias Ku h and Ch is ian Kubisch: analyzed and in e p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con en . Ma in Dichgans: concei ed and designed he s udy; analyzed and in e p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con en . Daniel I. Chasman and Ch is Co sapas: analyzed and in e - p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con- en . John-Anke Zwa , Nilesh J. Samani, and Aa no Palo ie: concei ed and designed he s udy; join ly supe ised esea ch; analyzed and in e - p e ed he da a; con ibu ed da a; d a ed/ e ised he manusc ip o con- en . All au ho s accep esponsibili y o conduc o esea ch and will gi e inal app o al. ACKNOWLEDGMENT P.P. was suppo ed by he Eu opean Commission FP7 p ojec no. 261123 (gEUVADIS). C.K. and H.G. we e unded by he Ge man Fede al Min- is y o Educa ion and Resea ch (BMBF) wi hin he amewo k o he Na ional Genome Resea ch Ne wo k (NGFN-Plus; g an s 01GS08120 and 01GS1103 [ o C.K.]) and he Deu sche Fo schungsgemeinscha (DFG). The Academy o Finland (g an 139795 o M.W.); he Folkhälsan Resea ch Founda ion ( o M.W.); he Medicinska Unde s öds ö eningen Li & Hälsa ( o M.W.); he O ion Fa mos Resea ch Founda ion ( o V. An ila); and he Helsinki Uni e si y Cen al Hospi al ( o M. Koi anen and V. A o). The Women ’ s Genome Heal h S udy (WGHS) is sup- po ed by HL043851 and HL080467 om he Na ional Hea , Lung, and Blood Ins i u e and CA047988 om he Na ional Cance Ins i u e, wi h collabo a i e scien i ic suppo and unding o geno yping p o ided by Amgen. Gene ic analyses o mig aine in WGHS ha e been suppo ed by NS061836 om he Na ional Ins i u e o Neu ological Diso de s and S oke. The No d-T øndelag Heal h S udy (The HUNT S udy) is a col- labo a ion be ween HUNT Resea ch Cen e (Facul y o Medicine, No - wegian Uni e si y o Science and Technology, NTNU), No d-T øndelag Coun y Council, Cen al No way Heal h Au ho i y, and he No wegian Ins i u e o Public Ins i u e o Public Heal h. The Young Finns S udy has been inancially suppo ed by he Academy o Finland: g an s 134309, 126925, 121584, 124282, 129378, 117787, and 41071, he Social Insu - ance Ins i u ion o Finland, Kuopio, Tampe e and Tu ku Uni e si y Hos- pi al Medical Funds (g an 9N035 o T.L.), Juho Vainio Founda ion, Paa o Nu mi Founda ion, Finnish Founda ion o Ca dio ascula Resea ch and Finnish Cul u al Founda ion, Tampe e Tube culosis Founda ion, and Emil Aal onen Founda ion. The au ho s g a e ully acknowledge he expe echnical assis ance in he s a is ical analyses by I ina Lisinen and Ville Aal o. TwinsUK: he s udy was unded by he Wellcome T us and Eu o- pean Communi y ’ s Se en h F amewo k P og amme (FP7/2007 – 2013). The s udy also ecei es suppo om he Na ional Ins i u e o Heal h Resea ch (NIHR) BioResou ce Clinical Resea ch Facili y and Biomedical Resea ch Cen e based a Guy ’ s and S Thomas ’ NHS Founda ion T us and King ’ s College London. SNP Geno yping was pe o med by The Wellcome T us Sange Ins i u e and Na ional Eye Ins i u e ia NIH/ CIDR. The au ho s hank hei unde s, win olun ee s, and TwinsUK eam. The LUMINA s udy is suppo ed by g an s ob ained om he Ne he lands O ganiza ion o he Heal h Resea ch and De elopmen (ZonMw) no. 90700217 and VIDI (ZonMw) no. 91711319 ( o G.M. T.); he Ne he lands O ganisa ion o Scien i ic Resea ch (NWO) VICI (918.56.602) and Spinoza (2009) g an s ( o M.D.F.); he 7 h F amewo k EU p ojec EUROHEADPAIN (no. 602633); and he Cen e o Medical Sys ems Biology (CMSB) es ablished in he Ne he lands Genomics Ini ia i e/Ne he lands O ganisa ion o Scien i ic Resea ch (NGI/ NWO), p ojec no. 050-060-409 ( o M.D.F. and A.M.J.M. .d.M.). Phe- no ype and geno ype da a collec ion in he Finnish Twin Coho has been suppo ed by he Wellcome T us Sange Ins i u e, ENGAGE—Eu opean Ne wo k o Gene ic and Genomic Epidemiology, FP7-HEALTH-F4- 2007, g an 201413, Na ional Ins i u e o Alcohol Abuse and Alcoholism (g an s AA-12502, AA-00145, and AA-09203 o R.J. Rose and AA15416 and K02AA018755 o D.M. Dick), and he Academy o Finland (g an s 100499, 205585, 118555, 141054, 265240, 263278, and 264146 o J.K.). Pheno ype and geno ype da a collec ion in NTR/NESDA was unded by he Ne he lands O ganiza ion o Scien i ic Resea ch (NWO: MagW/ZonMW g an s 904-61-090, 985-10-002, 904-61-193, 480-04- 004, 400-05-717, Addic ion-31160008 Middelg oo -911-09-032, Spino- zap emie 56-464-14192, Gees k ach p og am g an 10-000-1002), Cen- e o Medical Sys ems Biology (CMSB, NWO Genomics), Biobanking and Biomolecula Resou ces Resea ch In as uc u e (BBMRI — NL, 184.021.007), he VU Uni e si y ’ s Ins i u e o Heal h and Ca e Resea ch (EMGO 1 ), Neu oscience Campus Ams e dam (NCA), ENGAGE (HEALTH-F4-2007-201413); Eu opean Science Council (ERC Ad anced, 230374), Ru ge s Uni e si y Cell and DNA Reposi o y (NIMH U24 MH068457-06), he A e a Ins i u e o Human Gene ics, Sioux Falls, SD, he Na ional Ins i u es o Heal h (NIH, R01D0042157- 01A), Gene ic Associa ion In o ma ion Ne wo k (GAIN) o he Founda- ion o he US Na ional Ins i u es o Heal h (NIMH, MH081802), and by he G and Oppo uni y g an s 1RC2MH089951-01 and 1RC2MH089995-01 om he NIMH. NFBC1966 ecei ed inancial suppo om he Academy o Finland (p ojec g an s 104781, 120315, 129269, 1114194, 24300796, Cen e o Excellence in Complex Disease Gene ics and SALVE), Oulu Uni e si y Hospi al, Finland, Biocen e , Uni e si y o Oulu, Finland 75617, 24002054, Uni e si y o Oulu, Fin- land (g an s 24000692 and 24500283: Well-being and heal h: Resea ch in he No he n Finland Bi h Coho s 1966 and 1986, Pheno ypic and Genomic analyses). NIH/NHLBI NHLBI g an 5R01HL087679-02 h ough he STAMPEED p og am (1RL1MH083268-01), NHLBI Conso ium o Neu opsychia ic Phenomics Co-o dina ing Cen e (1R01HL087679-01), and NIH/NIMH (5R01MH63706-02), Uni ed S a es. ENGAGE p ojec and g an ag eemen HEALTH-F4-2007- 201413. Medical Resea ch Council (g an G1002319). The DNA ex ac ions, sample quali y con ols, biobank upkeep, and aliquo ing we e pe o med in he Na ional Public Heal h Ins i u e, Biomedicum Helsinki, Finland and suppo ed inancially by he Academy o Finland and Biocen um Helsinki. The au ho s hank Ms. Ou i To nwall and Ms. Min u Jussila (DNA biobanking). STUDY FUNDING This wo k was suppo ed by Academy o Finland (g an 251704 o A.P.), Sig id Juselius Founda ion ( o A.P.), SynSys ( o A.P.), he Wellcome T us (g an 098051 o A.P.), EU FP7-242167 ( o A.P.), NIH/RFA- HL-12-007, Genomic and Me abolomic P o iling o Finnish Familial Dyslipidemia Families ( o A.P.), he Sou h-Eas e n No way Regional Heal h Au ho i y (g an s 2010075 and 2011083 o B.S.W., L.M.J., and J.-A.Z.), he Resea ch Council o No way (g an 231187/F20 o B.S.W.), and he NIHR Leices e Ca dio ascula Biomedical Resea ch Uni and BHF ( o C.P.N.). N.J.S. holds a Chai unded by he B i ish Hea Founda ion and is an NIHR Senio In es iga o . Funding o s udy coho s and emaining au ho s a e lis ed in he acknowledgmen . The unde s had no ole in s udy design, da a collec ion and analysis, decision o publish, o p epa a ion o he manusc ip . DISCLOSURE Bendik S. Wins old has ecei ed esea ch suppo om he Resea ch Council o No way, he Sou h-Eas e n No way Regional Heal h Au ho - i y, and Fo sbe g’s and Auli Endowmen . Ch is ophe P. Nelson, Raine Malik, and Padh aig Go mley epo no disclosu es. Ve ne i An ila has ecei ed esea ch suppo om O ion Fa mos Resea ch Founda ion. Jason Vande Heiden has ecei ed unding o a el and/o speake hono a ia om New England Biolabs; and has ecei ed esea ch suppo om Na ional Lib a y o Medicine and he Uni ed S a es-Is ael Bina- ional Science Founda ion. Ka he ine S. Ellio epo s no disclosu es. Line M. Jacobsen is employed by As aZeneca. P ii Pal a has ecei ed 8 Neu ology: Gene ics ª 2015 Ame ican Academy o Neu ology. Unau ho ized ep oduc ion o his a icle is p ohibi ed. esea ch suppo om he Finnish Cul u al Founda ion. Naja Amin has ecei ed esea ch suppo om he Ne he lands B ain Founda ion. Boukje de V ies and Eija Hämäläinen epo no disclosu es. Tobias F eilinge has ecei ed unding o a el and/o speake hono a ia om Boeh inge Ingelheim and Alle gan; and has ecei ed esea ch suppo om Deu sche Fo schungsgemeinscha (DFG). M. A an Ik am has ecei ed unding o a el and/o speake hono a ia om Kaizo, L d.; has se ed on he edi o ial boa ds o Neu oepidemiology,PLoS One, and Jou nal o Alzheime Disease; and has ecei ed esea ch suppo om Janssen P e en ion Cen e , Ne he lands O ganiza ion o Heal h Resea ch and De elopmen , he Ne he lands Hea Founda ion, In e na- ionaal Pa kinson Fonds, In e na ionale S ich ing Alzheime Onde zoek, and he Alzheime Associa ion. Tho s en Kessle and Ma kku Koi anen epo no disclosu es. Lannie Lig ha has ecei ed esea ch suppo om EFIC-G ünen hal. Geo ge McMahon, Linda M. Pede sen, Ch is ina Willenbo g, and Hong-Hee Won epo no disclosu es. Jes Olesen has consul ed o Jannsen Pha maceu ical P oduc s; and has se ed on speak- e s’bu eaus o Alle gan. Ville A o has ecei ed unding o a el and/o speake hono a ia om Boeh inge Ingelheim, O ion, Mena ini, Mig aine T us , and Baye ; and has ecei ed esea ch suppo om he Finnish Medical Founda ion and Helsinki Uni e si y Cen al Hospi al. Themis ocles L. Assimes has se ed on he edi o ial boa d o F on ie s in Ca dio ascula Medicine; has consul ed and ecei ed esea ch suppo om Telome e Diagnos ics, Inc.; and has ecei ed esea ch suppo om NHLBI. S e an Blankenbe g, Do e I. Boomsma, Lynn Che kas, and Geo ge Da ey Smi h epo no disclosu es. S ephen E. Eps ein has ecei ed esea ch suppo om MedS a Hea and Vascula Ins i u e, MedS a WA Hospi al Cen e ; and holds s ock/s ock op ions and/o ecei es Boa d o Di ec o s Compensa ion o Ca dioCell. Jeane e E dmann epo s no disclosu es. Michel D. Fe a i has se ed on he edi o ial boa d o Cephalalgia; and has ecei ed esea ch suppo om he Ne he lands O ganiza ion o Scien i ic Resea ch (NWO), Eu opean Communi y, ZonMW, and he Du ch Hea Founda ion. Ha mu Göbel has se ed on Scien i ic Ad iso y Boa ds o Alle gan, Baye Vi al, and S . Jude Medical; has ecei ed unding o a el and/o speake hono a ia om Amgen, Alle gan, Ho mosan, Klos e au, MSD, Mundi- pha ma, S . Jude Medical, and Te a; has se ed on he edi o ial boa d o De Schme z, Pain Resea ch and T ea men ; has se ed on speake s’ bu eaus o Alle gan, Ho mosan, Klos e au, MSD, Mundipha ma, S . Jude Medical, and Te a; and has ecei ed esea ch suppo om S . Jude Medical. Alis ai S. Hall and Ma jo-Rii a Ja elin epo no disclosu es. Mikko Kallela has se ed on Ad iso y Boa ds o MSD and Alle gan; has ecei ed unding o a el and/o speake hono a ia om MSD, Alle gan, TEVA, No a is, and Genzyme; has ecei ed compen- sa ion o p oducing educa ional ma e ial om TEVA and Alle gan; has ecei ed esea ch suppo om Helsinki Uni e si y Cen al Hospi al; and holds s ock/s ock op ions and/o has ecei ed Boa d o Di ec o s com- pensa ion om Helsinki Headache Cen e . Jaakko Kap io has se ed on an Ad iso y Boa d o Copenhagen Uni e si y; has ecei ed unding o a el and/o speake hono a ia; and has consul ed o P ize L d. Seka Ka hi esan has se ed on scien i ic ad iso y boa ds o Regene on, Me ck, Eli Lilly, Aege ionm, Ca abasis, Ama in, and No a is; and has ecei ed esea ch suppo om Regene on, Aege ion, Me ck, NIH, and Fonda ion Leducq. Te ho Leh imäki epo s no disclosu es. Ru h McPhe son has se ed on he edi o ial boa d o A e ioscle osis, Th ombosis & Vascula Biology; and has ecei ed esea ch suppo om Canadian Ins i u es o Heal h Resea ch and Hea & S oke Founda ion o Canada. Win ied Mä z has se ed on Scien i ic Ad iso y Boa ds and speake s’bu eaus and consul ed o Aege ion Pha maceu icals, AMGEN, Danone Resea ch, Sano i/Genzyme, Ho mann LaRoche, MSD, Synage a, Eli Lilly, and BASF; has ecei ed unding o a el and/o speake hono a ia om Aege ion Pha maceu icals, AMGEN, Danone Resea ch, Sano i/ Genzyme, Ho mann LaRoche, MSD, Synage a, Eli Lilly, and BASF; has se ed on he edi o ial boa ds o Eu opean Hea Jou nal andJou nal o Labo a o y Medicine; has been employed by and holds s ock/s ock op ions and/o Boa d o Di ec o s compensa ion om Synlab Se ices GmbH; and has ecei ed esea ch suppo om Aege ion Pha maceu i- cals, AMGEN, Danone Resea ch, Sano i/Genzyme, Ho mann LaRoche, MSD, Synage a, Eli Lilly, BASF, Eu opean Union, Ge man Minis y o Resea ch, Ge man Minis y o Comme ce, and Wissenscha sini ia i e Obe hein. Dale R. Nyhol epo s no disclosu es. Ch is ophe J. O’Donnell is employed by and has ecei ed esea ch suppo om Na ional Ins i u es o Heal h. Lydia Quaye epo s no disclosu es. Daniel J. Rade has se ed on he scien i ic ad iso y boa ds o A e ioscle osis, Th ombosis & Vascula Biology; and has ecei ed esea ch suppo om NIH and he Leducq Founda ion. Olli Rai aka i and Robe Robe s epo no disclosu es. He ibe Schunke has se ed on scien i ic ad iso y boa ds, consul ed o , ecei ed unding o a el and/o hono a ia, and/o ecei ed esea ch suppo om As aZeneca, AMGEN, MSD SHARP & DOHME, Baye Vi al, Boeh inge Ingelheim, Med onic, No a is, P ize , Sano i-A en is, S . Jude, Bos on Scien i ic, and Daiichi Sankyo. Ma kus Schü ks has se ed on he edi o ial boa ds o The Jou nal o Headache and Pain, and BMC Neu ology; and consul s Baye Heal hCa e Pha maceu icals. Alexand e F.R. S ewa has se ed on he edi o ial boa d o F on ie s in Ca dio ascula Medicine — Ca dio ascula Gene ics . Gisela M. Te wind has ecei ed esea ch suppo om he Ne he lands O ganisa ion o Scien i ic Resea ch (NWO). Unnu Tho s einsdo i is an employee o deCODE gene ics/Amgen. A n M.J.M. an den Maagdenbe g and Co nelia an Duijn epo no disclosu es. Maija Wessman has ecei ed esea ch suppo om Folkhälsan Resea ch Founda ion, Academy o Finland, and Medicinska Unde s öds ö eningen Li och Hälsa. Tobias Ku h has se ed on edi o ial boa ds o BMJ and Cephalalgia; and has ecei ed esea ch suppo om he F ench Na ional Resea ch Agency and he Uni e si y o Bo deaux. Ch is ian Kubisch epo s no disclosu es. Ma in Dichgans has se ed on edi o ial boa ds o S oke, he In e na ional Jou nal o S oke , Ce eb o ascula Diseases , and Jou nal o Neu ochemis y ; has consul ed o Baye Vi al, Boeh inge Ingelheim, B is ol-Mye s Squibb, and Heel; and has ecei ed esea ch suppo om Wellcome T us , Eu opean Union, and Ge man Fede al Minis y o Educa ion and Resea ch. Daniel I. Chasman has se ed on he edi o ial boa d o A e ioscle osis, Th ombosis, and Vascula Biology; and ecei es publishing oyal ies o P o ein S uc u e: De e mina ion, Analysis, and Applica ions o D ug Disco e y (Ma cel Dekke , 2003). Ch is Co sapas has se ed on he edi o ial boa d o PLoS Gene ics; and has ecei ed esea ch suppo om NINDS, NIAID, and RE Child- en’s Conso ium. John-Anke Zwa epo s no disclosu es. Nilesh J. Samani has se ed on edi o ial boa ds o Ci cula ion: Ca dio ascula Gene - ics and Hea ; and has ecei ed esea ch suppo om B i ish Hea Foun- da ion and Na ional Ins i u e o Heal h Resea ch. Aa no Palo ie has been a membe o he P ize Gene ics Scien i ic Ad iso y Panel; has ecei ed a el expenses and/o hono a ia o lec u es o educa ional ac i i ies no unded by indus y; and has ecei ed esea ch suppo om he Finnish Academy, Eu opean Union NIH, NINDS, Juselius Founda ion, and he Finnish Founda ion o Ca dio ascula Resea ch. Go o Neu ology.o g/ng o ull disclosu e o ms. Recei ed Ap il 4, 2015. Accep ed in inal o m May 27, 2015. REFERENCES 1. Vos T, Flaxman AD, Nagha i M, e al. Yea s li ed wi h disabili y (YLDs) o 1160 sequelae o 289 diseases and inju ies 1990-2010: a sys ema ic analysis o he Global Bu den o Disease S udy 2010. Lance 2012; 380:2163–2196. 2. Headache Classi ica ion Commi ee o he In e na ional Headache Socie y (IHS). The In e na ional Classi ica ion o Headache Diso de s, 3 d edi ion (be a e sion). Ceph- alalgia 2013;33:629–808. 3. Cha les AC, Baca SM. Co ical sp eading dep ession and mig aine. Na Re Neu ol 2013;9:637–644. 4. Cha les A. Mig aine is no p ima ily a ascula diso de . Cephalalgia 2012;32:431–432. 5. Ashina M. Vascula changes ha e a p ima y ole in mig aine. Cephalalgia 2012;32:428–430. 6. Noseda R, Bu s ein R. Mig aine pa hophysiology: ana - omy o he igemino ascula pa hway and associa ed neu ological symp oms, co ical sp eading dep ession, sensi iza ion, and modula ion o pain. Pain 2013;154 (suppl 1):S44–S53. Neu ology: Gene ics 9