Genome-wide meta-analysis of 241,258 adults accounting for smoking behaviour identifies novel loci for obesity traits
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Recei ed 23 Jun 2016 |Accep ed 15 Feb 2017 |Published 26 Ap 2017
Genome-wide me a-analysis o 241,258 adul s
accoun ing o smoking beha iou iden ifies no el
loci o obesi y ai s
Anne E. Jus ice e al.#
Few genome-wide associa ion s udies (GWAS) accoun o en i onmen al exposu es, like
smoking, po en ially impac ing he o e all ai a iance when in es iga ing he gene ic
con ibu ion o obesi y- ela ed ai s. He e, we use GWAS da a om 51,080 cu en smoke s
and 190,178 nonsmoke s (87% Eu opean descen ) o iden i y loci influencing BMI and cen al
adiposi y, measu ed as wais ci cum e ence and wais - o-hip a io bo h adjus ed o BMI. We
iden i y 23 no el gene ic loci, and 9 loci wi h con incing e idence o gene-smoking in e ac ion
(GxSMK) on obesi y- ela ed ai s. We show consis en di ec ion o e ec o all iden ified
loci and significance o 18 no el and o 5 in e ac ion loci in an independen s udy sample.
These loci highligh no el biological unc ions, including esponse o oxida i e s ess,
addic i e beha iou , and egula o y unc ions emphasizing he impo ance o accoun ing o
en i onmen in gene ic analyses. Ou esul s sugges ha obacco smoking may al e he
gene ic suscep ibili y o o e all adiposi y and body a dis ibu ion.
Co espondence and eques s o ma e ials should be add essed o A.E.J. (email: [email p o ec ed]) o o L.A.C. (email: [email p o ec ed]).
#A ull lis o au ho s and hei a filia ions appea s a he end o he pape .
DOI: 10.1038/ncomms14977 OPEN
NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions 1
Recen genome-wide associa ion s udies (GWAS) ha e
desc ibed loci implica ed in obesi y, body mass index
(BMI) and cen al adiposi y. Ye mos s udies ha e igno ed
en i onmen al exposu es wi h possibly la ge impac s on he ai
a iance1,2. Va ian s ha exe gene ic e ec s on obesi y h ough
in e ac ions wi h en i onmen al exposu es o en emain
undisco e ed due o he e ogeneous main e ec s and s ingen
significance h esholds. Thus, s udies may miss gene ic a ian s
ha ha e e ec s in subg oups o he popula ion, such as
smoke s3.
I is o en no ed ha cu en ly smoking indi iduals display
lowe weigh /BMI and highe wais ci cum e ence (WC) as
compa ed o nonsmoke s4–6. Smoke s also ha e he smalles
fluc ua ions in weigh o e B20 yea s compa ed o hose who
ha e ne e smoked o ha e s opped smoking7,8. Also, hea y
smoke s (420 ciga e es pe day [CPD]) and hose ha ha e
smoked o mo e han 20 yea s a e a g ea e isk o obesi y han
non-smoke s o ligh o mode a e smoke s (o20 CPD)9,10. Men
and women gain weigh apidly a e smoking cessa ion and
many people in en ionally smoke o weigh managemen 11.I
emains unclea why smoking cessa ion leads o weigh gain o
why long- e m smoke s main ain weigh h oughou adul hood,
al hough s udies sugges ha obacco use supp esses appe i e12,13
o al e na i ely, smoking may esul in an inc eased me abolic
a e12,13. Iden i ying genes ha influence adiposi y and in e ac
wi h smoking may help us cla i y pa hways h ough which
smoking influences weigh and cen al adiposi y13.
A comp ehensi e s udy ha e alua es smoking in conjunc ion
wi h gene ic con ibu ions is wa an ed. Using GWAS da a om
he Gene ic In es iga ion o An h opome ic T ai s (GIANT)
Conso ium, we iden ified 23 no el gene ic loci, and 9 loci wi h
con incing e idence o gene-smoking in e ac ion (GxSMK) on
obesi y, assessed by BMI and cen al obesi y independen o o e all
body size, assessed by WC adjus ed o BMI (WCadjBMI) and
wais - o-hip a io adjus ed o BMI (WHRadjBMI). By accoun ing
o smoking s a us, we ocus bo h on gene ic a ian s obse ed
h ough hei main e ec s and GxSMK e ec s o inc ease ou
unde s anding o hei ac ion on adiposi y- ela ed ai s. These loci
highligh no el biological unc ions, including esponse o
oxida i e s ess, addic i e beha iou and egula o y unc ions
emphasizing he impo ance o accoun ing o en i onmen in
gene ic analyses. Ou esul s sugges ha smoking may al e he
gene ic suscep ibili y o o e all adiposi y and body a dis ibu ion.
Resul s
GWAS disco e y o e iew. We me a-analysed s udy-specific
associa ion esul s om 57 Hapmap-impu ed GWAS and 22
s udies wi h Me abochip, including up o 241,258 (87% Eu opean
descen ) indi iduals (51,080 cu en smoke s and 190,178
nonsmoke s) while accoun ing o cu en smoking (SMK)
(Me hods sec ion, Supplemen a y Fig. 1, Supplemen a y
Tables 1–4). Fo p ima y analyses, we conduc ed me a-analyses
ac oss ances ies and sexes. Fo seconda y analyses, we conduc ed
me a-analyses in Eu opean-descen s udies alone and sex-specific
me a-analyses (Tables 1–4, Supplemen a y Da a 1–6). We con-
side ed ou analy ical app oaches o e alua e he e ec s o
smoking on gene ic associa ions wi h adiposi y ai s (Fig. 1,
Me hods sec ion). App oach 1 (SNPadjSMK) examined gene ic
associa ions a e adjus ing o SMK. App oach 2 (SNPjoin )
conside ed he join impac o main e ec s adjus ed o SMK þ
in e ac ion e ec s14. App oach 3 ocused on in e ac ion e ec s
(SNPin ); App oach 4 ollowed up loci om App oach 1 o
in e ac ion e ec s (SNPsc een). Resul s om App oaches
1–3 we e conside ed genome-wide significan (GWS) wi h a
P- alueo5108while App oach 4 used Bon e oni
adjus men a e sc eening. Lead a ian s 4500 kb om
p e ious associa ions wi h BMI, WCadjBMI, and WHRadjBMI
we e conside ed no el. All associa ion esul s a e epo ed wi h
e ec es ima es o ien ed on he ai inc easing allele in he
cu en smoking s a um.
Ac oss he h ee adiposi y ai s, we iden ified 23 no el
associa ed gene ic loci (6 o BMI, 11 o WCadjBMI, 6 o
WHRadjBMI) and nine ha ing significan GxSMK in e ac ion
e ec s (2 o BMI, 2 o WCadjBMI, 5 o WHRadjBMI;
Fig. 1, Tables 1–4, Supplemen a y Da a 1–6). We p o ide a
comp ehensi e compa ison wi h p e iously-iden ified loci1,2
by ai in supplemen a y ma e ial (Supplemen a y Da a 7,
Supplemen a y No e 1).
Accoun ing o smoking s a us. Fo p ima y me a-analyses o
BMI (combined ances ies and sexes), 58 loci eached GWS in
App oach 1 (SNPadjSMK; Supplemen a y Da a 1, Supplemen a y
Figs 2 and 3), including wo no el loci nea SOX11,andSRRM1P2
(Table 1). Th ee mo e BMI loci we e iden ified using App oach 2
(SNPjoin ), including a no el locus nea CCDC93 (Supplemen a y
Figs 4 and 5). Fo WCadjBMI, 62 loci eached GWS o App oach
1 (SNPadjSMK) and wo mo e o App oach 2 (SNPjoin ),
including eigh no el loci nea KIF1B,HDLBP,DOCK3,
ADAMTS3,CDK6,GSDMC,TMEM38B and ARFGEF2 (Table 1,
Supplemen a y Da a 2, Supplemen a y Figs 2–5). Lead a ian s
nea PSMB10 om App oaches 1 and 2 ( s14178 and s113090,
espec i ely) a e 4500kb om a p e iously-iden ified
WCadjBMI-associa ed a ian ( s16957304); howe e , a e
condi ioning on he known a ian , ou signal is a enua ed
(P
Condi ional
¼3.02102and P
Condi ional
¼5.22103), indi-
ca ing ha his finding is no no el. Fo WHRadjBMI, 32 loci we e
iden ified in App oach 1 (SNPadjSMK), including one no el locus
nea HLA-C, wi h no addi ional loci in App oach 2 (SNPjoin ;
Table 1, Supplemen a y Da a 3, Supplemen a y Figs 2–5).
We used GCTA15 o iden i y loci om ou p ima y me a-
analyses ha ha bou mul iple independen SNPs (Me hods
sec ion, Supplemen a y Tables 5–7). Condi ional analyses
e ealed no seconda y signals wi hin 500 kb o ou no el
lead SNPs. Addi ionally, we pe o med condi ional associa ion
analyses o de e mine whe he ou no el a ian s we e
independen o p e ious GWAS loci wi hin 500 kb ha a e
associa ed wi h ela ed ai s o in e es . All BMI-associa ed SNPs
we e independen o p e iously iden ified GWS associa ions
wi h an h opome ic and obesi y- ela ed ai s. Se en no el loci
o WCadjBMI we e nea p e ious associa ions wi h ela ed
an h opome ic ai s. O hese, associa ion signals o s6743226
nea HDLBP, s10269774 nea CDK6, and s6012558 nea
ARFGEF2 we e a enua ed (P
Condi ional
41E5and bdec eased
by hal ) a e condi ioning on a leas one nea by heigh and
hip ci cum e ence adjus ed o BMI (HIPadjBMI) SNP,
bu associa ion signals emained independen o o he ela ed
SNP- ai associa ions. Fo WHRadjBMI, ou GWAS signal was
a enua ed by condi ioning on wo known heigh a ian s
( s6457374 and s2247056), bu emained significan in o he
condi ional analyses. Gi en high co ela ions among wais , hip
and heigh , hese esul s a e no su p ising.
Se e al addi ional loci we e iden ified o App oaches 1 and 2
in seconda y me a-analysis (Table 2, Supplemen a y Da a 1–6,
Supplemen a y Fig. 6). Fo BMI, 2 no el loci we e iden ified by
App oach 1, including 1 nea EPHA3 and 1 nea INADL. Fo
WCadjBMI, 2 no el loci we e iden ified nea RAI14 and PRNP.
Fo WHRadjBMI, fi e no el loci we e iden ified in seconda y
me a-analyses nea BBX, TRBI1,EHMT2,SMIM2 and EYA4.
A comp ehensi e summa y o nea by genes o all no el loci and
hei po en ial biological ele ance is a ailable in Supplemen a y
No e 2.
ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977
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Figu e 3 p esen s analy ical powe o App oaches 1 and 2
while Supplemen a y Table 8 and Supplemen a y Fig. 7 p esen
simula ion esul s o e alua e ype 1 e o (Me hods sec ion). A
hea map c oss- abula es P- alues o App oaches 1 and 2 along
wi h App oach 3 examining in e ac ion only (Supplemen a y
Fig. 8). We demons a e ha he wo app oaches yield alid ype
1 e o a es and ha App oach 1 can be mo e powe ul o find
associa ions gi en ze o o negligible quan i a i e in e ac ions,
whe eas App oach 2 is mo e e ficien in finding associa ions
when in e ac ion exis s.
Modifica ion o gene ic p edisposi ion by smoking. App oach
3 di ec ly e alua ed GxSMK in e ac ion (SNPin ; Table 3,
Supplemen a y Da a 1–6, Fig. 2, Supplemen a y Figs 9 and 10).
Fo p ima y me a-analysis o BMI, wo loci eached GWS
including a p e iously iden ified GxSMK in e ac ion locus nea
CHRNB4 ( e . 3), and a no el locus nea INPP4B. Bo h loci
exhibi GWS e ec s on BMI in smoke s and no e ec s in
nonsmoke s. Fo CHRNB4 (choline gic nico ine ecep o B4), he
a ian mino allele (G) exhibi s a dec easing e ec on BMI in
cu en smoke s (bsmk ¼0.047) bu no e ec in nonsmoke s
(bnonsmk ¼0.002). P e ious s udies iden ified nea by SNPs in
high LD associa ed wi h smoking (nonsynonymous, s16969968
in CHRNA5)3and a e ial calcifica ion ( s3825807, a missense
a ian in ADAMTS7)16. Condi ioning on hese a ian s
a enua ed ou in e ac ion e ec bu did no elimina e i
(Supplemen a y Table 7), sugges ing a complex ela ionship
be ween smoking, obesi y, hea disease, and gene ic a ian s in
his egion. Impo an ly, he CHRNA5-CHRNA3-CHRNB4 gene
clus e has been associa ed wi h lowe BMI in cu en smoke s3,
bu wi h highe BMI in ne e smoke s3, e idence suppo ing he
lack o associa ion in nonsmoke s as well as a lack o p e ious
GWAS findings on 15q25 (Supplemen a y Da a 8)1. The
CHRNA5-CHRNA3-CHRNB4 genes encode he nico inic
ace ylcholine ecep o (nAChR) subuni s a3, a5 and b4, which
a e exp essed in he cen al ne ous sys em17. Nico ine has
di e ing e ec s on he body and b ain, causing changes in
me abolism and eeding beha iou s18. These findings sugges
smoking exposu e may modi y gene ic e ec s on 15q24-25 o
influence smoking- ela ed diseases, such as obesi y, h ough
dis inc pa hways.
In p ima y me a-analyses o WCadjBMI, one no el GWS locus
(nea GRIN2A) wi h opposi e e ec di ec ions by smoking s a us
was iden ified o App oach 3 (SNPin ; Table 3, Supplemen a y
Da a 2, Fig. 2, Supplemen a y Figs 9 and 10). The T allele o
s4141488 inc eases WCadjBMI in cu en smoke s and dec eases
i in nonsmoke s (bsmk ¼0.037, bnonsmk ¼0.015). In
seconda y me a-analysis o Eu opean women-only, we iden ified
an in e ac ion be ween s6076699, nea PRNP, and SMK on
WCadjBMI (Table 4, Supplemen a y Da a 5, Supplemen a y
Fig. 6), a locus also iden ified in App oach 2 (SNPjoin ) o
Eu opean women. The majo allele, A, has a posi i e e ec on
cu en smoke s as compa ed o a weake and nega i e e ec
on WC in nonsmoke s (bsmk ¼0.169, bnonsmk ¼0.070),
sugges ing why his a ian emained unde ec ed in p e ious
GWAS o WCadjBMI (Supplemen a y Da a 8).
App oach 4 (SNPsc een; Fig. 1, Me hods sec ion) e alua ed
GxSMK in e ac ions a e sc eening SNPadjSMK esul s ( om
Tes ing o In e ac ion o SNP wi h
Cu en SMK
Tes ing o SNP accoun ing o
cu en SMK
WHRadjBMI: 45 / 6 WCadjBMI: 76 / 11BMI: 68 / 6
App oach 4.b
Tes SNP
adjus ing o
SMK
P
SNPadjSMK
<5E
–8
App oach 1
To al numbe o signi ican loci/numbe o no el
In e ac ion o SNP wi h
cu en SMK
E ec o SNP
accoun ing o cu en SMK
App oach 1 App oach 2 App oach 3 App oach 4
0
0
App oach 3
App oach 2
Tes SNP +
in e ac ion wi h
SMK
P
SNPjoin
<5E
–8
Tes SNP
in e ac ion wi h
SMK
P
SNPin
<5E
–8
Sc een
app oach 1
esul s
P
SNPadjSMK
<5E
–8
Tes selec ed
SNP in e ac ion
P
SNPin
< 0.05/# loci
T ai
BMI
WCadjBMI
WHRadjBMI 44 / 5
72 / 9
65 / 4 57 / 2
66 / 5
24 / 2
2 / 2
2 / 1 1 / 0
5 / 0
To al non-o e lapping loci
App oach 4.a
Figu e 1 | Summa y o s udy design and esul s. App oach 1 uses bo h SNP and SMK in he associa ion model. App oaches 2 and 3 use he SMK-s a ified
me a-analyses. App oach 4 sc eens loci based on App oach 1, hen uses SMK-s a ified esul s o iden i y loci wi h significan in e ac ion e ec s
(Me hods sec ion).
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NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions 3
App oach 1) using Bon e oni-co ec ion (Me hods sec ion,
Tables 3–4, Supplemen a y Da a 1–6). We iden ified wo SNPs,
nea LYPLAL1 and RSPO3, wi h significan in e ac ion; bo h ha e
p e iously published main e ec s on an h opome ic ai s. These
loci exhibi e ec s on WHRadjBMI in nonsmoke s, bu no in
smoke s (Fig. 2). In seconda y me a-analyses, we iden ified
h ee known loci wi h significan GxSMK in e ac ion e ec s
on WHRadjBMI nea MAP3K1,HOXC4-HOXC6 and JUND
(Table 4, Supplemen a y Da a 3 and 6). We iden ified s1809420,
nea CHRNA5-CHRNA3-CHRNB4, o BMI in he men-only,
combined-ances ies me a-analysis (Supplemen a y Da a 1).
Powe calcula ions demons a e ha App oach 4 has inc eased
powe o iden i y SNPs ha show (i) an e ec in one s a um
(smoke s o nonsmoke s) and a less p onounced bu conco dan
e ec in he o he s a um, o (ii) an e ec in he la ge
nonsmoke s a um and no e ec in smoke s (Fig. 3). In con as ,
App oach 3 has inc eased powe o SNPs ha show (i) an e ec
in he smalle smoke s a um and no e ec in nonsmoke s, o
(ii) an opposi e e ec be ween smoke s and nonsmoke s (Fig. 3).
Ou findings o bo h app oaches ag ee wi h hese powe
p edic ions, suppo ing using bo h analy ical app oaches o
iden i y GxSMK in e ac ions.
En ichmen o gene ic e ec s by smoking s a us. When exam-
ining he smoking specific e ec s o BMI and WCadjBMI loci in
ou me a-analyses, no significan en ichmen o gene ic e ec s by
smoking s a us we e no ed. (Fig. 2, Supplemen a y Figs 11 and 12).
Howe e , ou esul s o WHRadjBMI we e en iched o loci
wi h a s onge e ec in nonsmoke s as compa ed o smoke s,
wi h 35 o 45 loci displaying nume ically la ge e ec s in
nonsmoke s (P
binomial
¼1.2104).
We calcula ed he a iance explained by subse s o SNPs
selec ed on 15 significance h esholds o App oach 1 om
P
SNPadjSMK
¼1108 o P
SNPadjSMK
¼0.1 (Supplemen a y
Table 9, Fig. 4). Di e ences in a iance explained be ween smoke s
and nonsmoke s we e significan (P
RsqDi
o0.003¼0.05/15,
Bon e oni-co ec ed o 15 h esholds) o BMI a each
h eshold, wi h mo e a iance explained in smoke s. Fo
WCadjBMI, he di e ence was significan o SNP se s beginning
wi h P
SNPadjSMK
Z3.16104, and o WHRadjBMI a
P
SNPadjSMK
Z1106. In con as o BMI, SNPs om App oach
1 explained a g ea e p opo ion o he a iance in nonsmoke s
o WHRadjBMI. Di e ences in a iance explained we e g ea es
o BMI (di e ences anged om 1.8 o 21% o smoke s)
and lowes o WHRadjBMI ( anging om 0.3 o 8.8% o
nonsmoke s).
These esul s sugges ha smoking may inc ease gene ic
suscep ibili y o o e all adiposi y, bu a enua e gene ic e ec s
on body a dis ibu ion. This con as is conco dan wi h
pheno ypic obse a ions o highe o e all adiposi y and lowe
cen al adiposi y in smoke s4,6,7. Addi ionally, smoking inc eases
oxida i e s ess and gene al inflamma ion in he body19 and may
exace ba e weigh gain20. Many genes implica ed in BMI a e
in ol ed in appe i e egula ion and eeding beha iou 1. Fo wais
ai s, ou esul s adjus ed o BMI likely highligh dis inc
pa hways h ough which smoking al e s gene ic suscep ibili y o
body a dis ibu ion. O e all, ou esul s indica e ha mo e loci
emain o be disco e ed as mo e a iance in he ai can be
explained as we d op he h eshold o significance.
Func ional o biological ole o no el loci. We conduc ed
ho ough sea ches o he li e a u e and publicly a ailable
bioin o ma ics da abases o unde s and he unc ional ole o all
genes wi hin 500kb o ou lead SNPs. We sys ema ically
explo ed he po en ial ole o ou no el loci in a ec ing gene
exp ession bo h wi h and wi hou accoun ing o he influence o
smoking beha iou (Me hods sec ion, Supplemen a y No e 3,
Supplemen a y Tables 10–12).
We ound he majo i y o no el loci a e nea s ong candida e
genes wi h biological unc ions simila o p e iously iden ified
adiposi y- ela ed loci, including egula ion o body a /weigh ,
angiogenesis/adipogenesis, glucose and lipid homeos asis, gene al
g ow h and de elopmen . (Supplemen a y No es 1 and 3).
We iden ified s17396340 o WCadjBMI (App oaches 1 and 2),
an in onic a ian in he KIF1B gene. This a ian is associa ed
wi h exp ession o KIF1B in whole blood wi h and wi hou
accoun ing o SMK (GTeX and Supplemen a y Tables 10 and 12)
and is highly exp essed in he b ain21. Knockou and mu an o ms
o KIF1B in mice esul ed in mul iple b ain abno mali ies,
including hippocampus mo phology22, a egion in ol ed in
( ood) memo y and cogni ion23. Va ian s17396340 is
associa ed wi h exp ession le els o ARSA in LCL issue. Human
adipocy es exp ess unc ional ARSA, which u ns dopamine sul a e
in o ac i e dopamine. Dopamine egula es appe i e h ough lep in
INPP4B*
CCDC39
CHRNB4*
SRRM1P2
SOX11 ADAMTS3
KIF1B
LYPLAL1 ¥
RSPO3 ¥
TMEM38B
HLA-C
HDLBP
CDK6
ARFGEF2
DOCK3
GRIN2A*
0.08
0.1
0.06
0.04
0.02
E ec on BMI (SD pe allele)
0
–0.02
–0.04
0.08
–0.01
0
0.01
0.02
0.03
0.04
0.05
0.06
0.04
0.02
E ec on WCadjBMI (SD pe allele) E ec on WHRadjBMI (SD pe allele)
0
–0.02
–0.04
Smoke s
Non-smoke s
Smoke s
Non-smoke s
Smoke s
Non-smoke s
abc
Figu e 2 | Fo es plo o no el and GxSMK loci s a ified by smoking s a us. Es ima ed e ec s (b±95% CI) o smoke s (Nup o 51,080) and
nonsmoke s (Nup o 190,178 ) pe isk allele o (a) BMI, (b) WCadjBMI and (c) WHRadjBMI o no el loci om App oaches 1 and 2 (SNPadjSMK and
SNPjoin , espec i ely) and all loci om App oaches 3 and 4 (SNPin and SNPsc een) iden ified in he p ima y me a-analyses. Loci a e o de ed by g ea e
magni ude o e ec in smoke s compa ed o nonsmoke s and labelled wi h he nea es gene. Fo he locus nea TMEM38B, s9409082 was used o e ec
es ima es in his plo . ( loci iden ified o App oach 4, *loci iden ified o App oach 3).
ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977
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and adiponec in le els, sugges ing a ole o ARSA in egula ing
appe i e24.
Exp ession o CD47 (CD47 molecule), nea s670752 o
WHRadjBMI (App oach 1, women-only), is significan ly
dec eased in obese indi iduals and nega i ely co ela ed wi h
BMI, WC and Hip ci cum e ence25. Con e sely, in mouse
models, CD47-deficien mice show dec eased weigh gain on
high- a die s, inc eased ene gy expendi u e, imp o ed glucose
p ofile and dec eased inflamma ion26.
Se e al no el loci ha bou genes in ol ed in unique biological
unc ions and pa hways including addic i e beha iou s and
esponse o oxida i e s ess. These po en ial candida e genes
nea ou associa ion signals a e highly exp essed in ele an
issues o egula ion o adiposi y and smoking beha iou
( o example, b ain, adipose issue, li e , lung and muscle;
Supplemen a y No e 2, Supplemen a y Table 10).
The CHRNA5-CHRNA3-CHRNB4 clus e is in ol ed in
he eNOS signalling pa hway (Ingenui y KnowledgeBase,
h p://www.ingenui y.com) ha is key o neu alizing eac i e
oxygen species in oduced by obacco smoke and obesi y27.
Dis up ion o his pa hway has been associa ed wi h
dys egula ion o adiponec in in adipocy es o obese mice,
implica ing his pa hway in downs eam e ec s on weigh
egula ion27,28. This finding is especially impo an due o he
compounded s ess adiposi y places on he body as i inc eases
ch onic oxida i e s ess i sel 28.INPP4B has been implica ed in
he egula ion o he PI3K/Ak signalling pa hway29 ha is
impo an o cellula g ow h and p oli e a ion, bu also eNOS
signalling, ca bohyd a e me abolism, and angiogenesis30.
GRIN2A, nea s4141488, con ols long- e m memo y
and lea ning h ough egula ion and e ficiency o synap ic
ansmission31 and has been associa ed wi h he oin addic ion32.
Nico ine inc eases he exp ession o GRIN2A in he p e on al
co ex in mu ine models33. The e a e no es ablished ela ionships
be ween GRIN2A and obesi y- ela ed pheno ypes in he li e a u e,
ye meman ine and ke amine, pha macological an agonis s o
0.00010
0.0
0.2
0.4
0.6
0.8
1.0
Powe
0.00005
0.00070
0.0034 0.0017 0.0000 0.0017 0.0034 0.0034 0.0017 0.0000 0.0017 0.0034
0.00035 0.00000 0.00035 0.00070 0.00070 0.00035 0.00000 0.00035 0.00070
Opposi e di ec ion Consis en di ec ion
0.00000 0.00005 0.00010 0.00010
0.0
0.2
0.4
0.6
0.8
1.0
Powe
0.0
0.2
0.4
0.6
0.8
1.0
Powe
0.0
0.2
0.4
0.6
0.8
1.0
Powe
0.0
0.2
0.4
0.6
0.8
1.0
Powe
0.0
0.2
0.4
0.6
0.8
1.0
Powe
0.00005 0.00000 0.00005 0.00010
R2
SMK = 0.01%
R2
SMK = 0.07%
R2
NONSMK = 0.01%
R2
NONSMK = 0.07%
R2
SMK = 0.34% R2
NONSMK = 0.34%
R2
NONSMK Opposi e di ec ion Consis en di ec ion
R2
SMK
Opposi e di ec ion Consis en di ec ion
R2
NONSMK Opposi e di ec ion Consis en di ec ion
R2
SMK
Opposi e di ec ion Consis en di ec ion
R2
NONSMK Opposi e di ec ion Consis en di ec ion
R2
SMK
App oach 1 (PadjSMK < 5 x 10–8)
App oach 3 (Pin < 5 x 10–8)
App oach 2 (PJoin 2d < 5 x 10–8)
App oach 4 (PadjSMK < 5 x 10–8 → Pin <0.05 / # loci)
ab
cd
e
Figu e 3 | Powe compa ison ac oss App oaches. Shown is he powe o iden i y adjus ed (App oach 1, dashed black lines), join (App oach 2,
do ed g een lines) and in e ac ion (App oach 3 and 4, solid magen a and o ange lines) e ec s o a ious combina ions o SMK- and NonSMK-specific
e ec s and assuming 50,000 smoke s and 180,000 nonsmoke s. Fo (a,c,e), he e ec in smoke s was fixed a a small (R2
SMK ¼0.01%, simila o he
ealis ic NUDT3 e ec on BMI), medium (R2
SMK ¼0.07%, simila o he ealis ic BDNF e ec on BMI) o la ge (R2
SMK ¼0.34%, simila o he ealis ic FTO
e ec on BMI) gene ic e ec , espec i ely, and a ied in nonsmoke s. Fo (b,d, ), he e ec in nonsmoke s was fixed o he small, medium and la ge BMI
e ec s, espec i ely, and a ied in smoke s.
NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977 ARTICLE
NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions 5
GRIN2A ac i i y34,35, a e implica ed in ea men o obesi y-
associa ed diso de s, including binge-ea ing diso de s and
mo bid obesi y (ClinicalT ials.go iden ifie s: NCT00330655,
NCT02334059, NCT01997515, NCT01724983). Meman ine is
unde clinical in es iga ion o ea men o nico ine dependence
(ClinicalT ials.go iden ifie s: NCT01535040, NCT00136786 and
NCT00136747). While ou lead SNP is no wi hin a cha ac e ized
gene, s4141488 and a ian s in high LD ( 240.7) a e wi hin
ac i e enhance egions o se e al issues, including li e , e al leg
muscle, smoo h s omach and in es inal muscle, co ex and se e al
emb yonic and plu ipo en cell ypes (Supplemen a y No e 2),
and he e o e may ep esen an impo an egula o y egion o
nea by genes like GRIN2A.
In seconda y me a-analysis o Eu opean women-only, we
iden ified a significan GxSMK in e ac ion o s6076699 on
WCadjBMI (Table 4, Supplemen a y Da a 4, Supplemen a y
Fig. 6). This SNP is 100kb ups eam o PRNP (p ion p o ein),
a signalling ansduce in ol ed in mul iple biological p ocesses
ela ed o he ne ous sys em, immune sys em, and o he cellula
unc ions (Supplemen a y No e 2)36. Al e na e o ms o he
oligome s may o m in esponse o oxida i e s ess caused by
coppe exposu e37. Coppe is p esen in ciga e e smoke and
Va iance explained o BMI
in smoke s and nonsmoke s
The di e ence in he a iance explained o BMI
in smoke s and nonsmoke s
0.30
0.25
0.20
0.15
To al explained a iance
To al explained
a iance smoke s s nonsmoke s
0.10
0.05
0.00
0.30
0.25
0.20
0.15
0.10
0.05
0.00
1E–08 1E–06
P adjus ed o smoking h eshold
1E–04 1E–02 1E–08 1E–06
P adjus ed o smoking h eshold
1E–04 1E–02
1E–08 1E–06
P adjus ed o smoking h eshold
1E–04 1E–02 1E–08 1E–06
P adjus ed o smoking h eshold
1E–04 1E–02
1E–08 1E–06
P ad
j
us ed o smokin
g
h eshold
1E–04 1E–02 1E–08 1E–06
P ad
j
us ed o smokin
g
h eshold
1E–04 1E–02
Smoke s
Nonsmoke s
Va iance explained o WCadjBMI
in smoke s and nonsmoke s
The di e ence in he a iance explained o WCadjBMI
in smoke s and nonsmoke s
0.30
0.25
0.20
0.15
To al explained a iance
To al explained
a iance nonsmoke s s smoke s
0.10
0.05
0.00
0.30
0.25
0.20
0.15
0.10
0.05
0.00
Va iance explained o WHRadjBMI
in smoke s and nonsmoke s
The di e ence in he a iance explained o WHRadjBMI
in smoke s and nonsmoke s
0.30
0.25
0.20
0.15
To al explained a iance
To al explained
a iance smoke s s nonsmoke s
0.10
0.05
0.00
0.5
0.4
0.3
0.2
0.1
0.0
ab
cd
e
Smoke s
Nonsmoke s
Smoke s
Nonsmoke s
Figu e 4 | S a um specific es ima es o a iance explained. To al smoking s a us-specific explained a iance (±s.e.) by SNPs mee ing a ying
h esholds o o e all associa ion in App oach 1 (SNPadjSMK) and he di e ence be ween he p opo ion o a iance explained be ween smoke s and
nonsmoke s o hese same se s o SNPs in BMI (a,b), WCadjBMI (c,d), and o WHRadjBMI (e, ).
ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977
6NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions
ele a ed in he se um o smoke s, bu is wi hin sa e anges38,39.
Ano he gene nea s6076699, SLC23A2 (Solu e Ca ie Family 23
(Asco bic Acid T anspo e ), Membe 2), is essen ial o he
up ake and anspo o Vi amin C, an impo an nu ien o
DNA and cellula epai in esponse o oxida i e s ess bo h
di ec ly and h ough suppo ing he epai o Vi amin E a e
exposu e o oxida i e agen s40,41. SLC23A2 is p esen in he
ad enal glands and mu ine models indica e ha i plays an
impo an ole in egula ing dopamine le els42. This egion is
associa ed wi h success in smoking cessa ion and is implica ed in
addic i e beha iou s in gene al43,44. Ou ag SNP is loca ed
wi hin an ac i e enhance egion (ma ked by open ch oma in
ma ks, DNAse hype sen i i y, and ansc ip ion ac o binding
mo i s); his egula o y ac i i y appea s issue specific
(sex-specific issues and lungs; HaploReg and UCSC Genome
B owse ).
Nico inamide mononucleo ide adenylyl ans e ease (NMNAT1),
ups eam o WCadjBMI a ian s17396340, is esponsible o
he syn hesis o NAD om ATP and NMN45. NAD is necessa y
o cellula epai ollowing oxida i e s ess. Up egula ion o
NMNAT p o ec s agains damage caused by eac i e oxygen
species in he b ain, specifically he hippocampus46. Also o
WCadjBMI, bo h CDK6, nea SNP s10269774, and FAM49B,
nea SNP s6470765, a e a ge s o he BACH1 ansc ip ion
ac o , in ol ed in cellula esponse o oxida i e s ess and
managemen o he cell cycle47.
Influence o no el loci on ela ed ai s. In a look-up in exis ing
GWAS o smoking beha iou s (E e /Ne e , Cu en /No -
Cu en , Smoking Quan i y (SQ))48 (Supplemen a y Da a 8),
eigh o ou 26 SNPs we e nominally associa ed wi h a leas one
smoking ai . A e mul iple es co ec ion (P
Reg ession
o0.05/
26 ¼0.0019), only one SNP emains significan : s12902602,
iden ified o App oaches 2 (SNPjoin ) and 3 (SNPin ) o BMI,
showed associa ion wi h SQ (P¼1.45 109).
We conduc ed a sea ch in he NHGRI-EBI GWAS Ca alog49,50
o de e mine i any o ou newly iden ified loci a e in high LD
wi h a ian s associa ed wi h ela ed ca diome abolic and
beha iou al ai s o diseases. O he se en no el BMI SNPs,
only s12902602 was in high LD ( 240.7) wi h SNPs p e iously
associa ed wi h smoking- ela ed ai s ( o example, nico ine
dependence), lung cance , and ca dio ascula diseases ( o
example, co ona y hea disease; Supplemen a y Table 13).
O he 12 no el WCadjBMI SNPs, 5 we e in high LD wi h
p e iously epo ed GWAS a ian s o mean pla ele
olume, heigh , in an leng h, and melanoma. O he six no el
WHRadjBMI SNPs, h ee we e nea se e al p e iously associa ed
a ian s, including ca diome abolic ai s ( o example, LDL
choles e ol, iglyce ides and measu es o enal unc ion).
Gi en high pheno ypic co ela ion be ween WC and WHR
wi h heigh , and es ablished sha ed gene ic associa ions ha
o e lap ou adiposi y ai s and heigh 1,2,51 we expec c oss- ai
associa ions be ween ou no el loci and heigh . The e o e,
we conduc ed a look-up o all o ou no el SNPs o
iden i y o e lapping associa ion signals (Supplemen a y Da a 8).
No no el BMI loci we e significan ly associa ed wi h heigh
(P
Reg ession
o0.002(0.05/24) SNPs). Howe e , he e a e addi ional
a ian s ha may be associa ed wi h heigh , bu no p e iously
epo ed in GWAS examining heigh , including wo o
WHRadjBMI nea EYA4 and TRIB1, and wo o WCadjBMI
nea KIF1B and HDLBP (P
Reg ession
o0.002).
Finally, as smoking has a nega i e (weigh dec easing) e ec on
BMI, i is likely ha smoking-associa ed gene ic a ian s ha e an
e ec on BMI in cu en smoke s. The e o e, we expec ed ha
smoking-associa ed SNPs exhibi some in e ac ion wi h smoking
on BMI. We looked up published smoking beha iou SNPs49,50,
10 a ian s in 6 loci, in ou own esul s. Two a ian s eached
nominal significance (P
SNPin
o0.05) o GxSMK in e ac ion
on BMI (Supplemen a y Table 14), bu only one eached
Bon e oni-co ec ed significance (Po0.005). No smoking-
associa ed SNPs exhibi ed GxSMK in e ac ion. The e o e, we
did no see a s ong en ichmen o low in e ac ion P alues
among p e iously iden ified smoking loci.
Valida ion o no el loci. We pu sued alida ion o ou no el and
in e ac ion SNPs in an independen s udy sample o up o
119,644 Eu opean adul s om he UK Biobank s udy (Tables 1–4,
Supplemen a y Table 15, Supplemen a y Fig. 9). We ound con-
sis en di ec ions o e ec s in smoking s a a ( o App oaches 2
and 3) and in SNPadjSMK esul s (App oach 1) o each locus
examined (Supplemen a y Fig. 13). Fo BMI, h ee SNPs we e
no GWS (P
SNPadjSMK
,P
SNPjoin
,P
SNPIn
45E8) ollowing me a-
analysis wi h ou GIANT esul s: s12629427 nea EPAH3
(App oach 1); s1809420 wi hin a known locus nea ADAMTS7
(App oach 4) emained significan o in e ac ion, bu no o
SNPadjSMK; and s336396 nea INPP4B (App oach 3). Fo
WCadjBMI, 3 SNPs we e no GWS (P
SNPadjSMK
,P
SNPjoin
,
P
SNPIn
45E8) ollowing me a-analysis wi h ou esul s:
Table 1 | Summa y o associa ion esul s o no el loci eaching genome-wide significance in App oach (App) 1 (P
SNPadjSMK
o5E8) o App oach 2 (P
SNPjoin
o5E8) o ou p ima y me a-analysis in combined ances ies and combined sexes.
App Ma ke Ch :Pos
(hg19)
Nea es
Gene
NEAF Alleles
E/O
Smoke s Non-smoke s Main and in e ac ion e ec s GIANT þUKBB
bP- alue bP- alue b
adj
P
SNPadjSMK
P
SNPin
P
SNPjoin
P
SNPadjSMK
P
SNPin
P
SNPjoin
BMI
1,2 s10929925 2:6155557 SOX11 225,067 0.55 C/A 0.019 7.80E03 0.02 8.40E08 0.020 1.1E09 8.2E 01 1.6E08 1.5E 13 4.5E 01 9.8E 13
1 s6794880 3:84451512 SRRM1P2 186,968 0.85 A/G 0.025 2.30E02 0.027 3.90E06 0.028 4.3E08 8.5E01 1.8E06 4.9E09 4.5E01 9.7E08
2 s13069244 3:180441172 CCDC39 233,776 0.08 A/G 0.061 1.80E05 0.031 6.60E05 0.035 1.2E07 4.6E02 3.5E 08 6.1E 10 1.1E 02 9.6E 11
WCadjBMI
1,2 s17396340 1:10286176 KIF1B 206,485 0.14 A/G 0.016 1.40E01 0.035 4.70E10 0.028 3.0E08 9.8E02 9.1E10 1.0E11 2.9E02 1.5E13
1,2 s6743226 2:242236972 HDLBP 200,666 0.53 C/T 0.018 1.30E02 0.023 2.60E09 0.022 1.2E 10 5.5E01 5.8E10 6.7E12 7.0E01 2.8E11
1 s4378999 3:51208646 DOCK3 156,566 0.13 T/A 0.035 1.30E02 0.035 1.30E06 0.036 4.1E08 9.7E 01 4.1E 07 7.6E 11 5.3E 01 3.2E 10
1,2 s7697556 4:73515313 ADAMTS3 206,017 0.49 T/C 0.004 6.30E01 0.025 7.30E11 0.021 5.2E09 6.7E 03 7.6E 10 5.4E 19 1.9E 02 2.7E 19
1 s10269774 7:92253972 CDK6 157,552 0.34 A/G 0.024 6.60E 03 0.023 1.10E 06 0.023 2.9E 08 8.8E 01 1.6E 07 2.9E 10 7.7E 01 2.1E 09
1 s6470765 8:130736697 GSDMC 157,450 0.76 A/C 0.032 1.90E03 0.023 1.70E05 0.026 4.8E08 4.3E01 9.5E07 2.5E 12 8.9E 01 9.0E 11
2 s9408815 9:108890521 TMEM38B 156,427 0.75 C/G 0.012 2.30E01 0.03 4.20E09 0.026 2.3E08 8.5E02 1.7E08 1.2E 11 3.0E01 2.8E11
1 s9409082 9:108901049 157,785 0.76 C/T 0.017 8.10E 02 0.029 2.60E 08 0.027 1.5E 08 2.7E 01 4.6E 08 9.5E 12 6.6E 01 6.5E 11
1 s6012558 20:47531286 ARFGEF2 208,004 0.41 A/G 0.026 5.40E04 0.018 6.50E06 0.020 1.9E 08 3.3E01 1.3E07 1.5E 09 7.0E02 3.0E09
WHRadjBMI
1,2 s1049281 6:31236567 HLA-C149,285 0.66 C/T 0.022 1.30E02 0.027 2.00E08 0.025 2.2E09 5.6E01 5.3E09 1.2E 18 8.3E01 1.8E10
Adj, adjus ed o smoking; app, app oach; in , in e ac ion; ch , ch omosome; EAF, e ec allele equency; E/O, e ec /o he ; Pos, posi ion (bp). Significan P- alues ha each genome-wide significance
(Po5108) h eshold a e in bold.
NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977 ARTICLE
NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions 7
s1545348 nea RAI14 (App oach 1); s4141488 nea GRIN2A
(App oach 3); and s6012558 nea PRNP (App oach 3). Fo
WHRadjBMI, only 1 SNP om App oach 4 was no significan
ollowing me a-analysis wi h ou esul s: s12608504 nea
JUND emained GWS o SNPadjSMK, bu was only nominally
significan o in e ac ion (P
SNPin
¼0.013).
Challenges in accoun ing o en i onmen al exposu es in GWAS.
A possible limi a ion o ou s udy may be he defini ion and
ha moniza ion o smoking s a us. We chose o s a i y on cu en
smoking s a us wi hou conside a ion o ype o smoking
( o example, ciga e e, pipe) o wo easons. Fi s , ocusing on
weigh alone, o me smoke s end o e u n o hei expec ed
weigh quickly ollowing smoking cessa ion7,13,52. Second, his
defini ion allowed us o maximize sample size, as many
pa icipa ing s udies only had cu en smoking s a us a ailable.
Howe e , WC and WHR may no beha e in he same manne
as weigh and BMI wi h o me smoke s e aining excess a
a ound hei wais . Thus, esul s may di e wi h al e na i e
ha moniza ion o smoking exposu e.
Ano he limi a ion may be po en ial bias in ou e ec es ima es
when adjus ing o a co ela ed co a ia e ( o example, collide
bias)53. This phenomenon is o pa icula conce n when he
co ela ion be ween he ou come and he co a ia e is high and
when significan gene ic associa ions occu wi h bo h ai s in
opposi e di ec ions. Ou analyses adjus ed bo h WC and WHR
o BMI. WHR has a co ela ion o 0.49 wi h BMI, while WC has
a co ela ion o 0.85 ( e . 53). Using p e iously published esul s
o BMI, WCadjBMI and WHRadjBMI, we find h ee no el loci
o WCadjBMI (nea DOCK3, ARFGEF2 and TMEM38B) and
wo o WHRadjBMI (nea EHMT2 and HLA-C; Supplemen a y
Da a 8) wi h nominally significan associa ions wi h BMI and
opposi e di ec ions o e ec . A hese loci, he gene ic e ec
es ima es should be in e p e ed wi h cau ion. Addi ionally, we
adjus ed o SMK in App oach 1 (SNPadjSMK). Howe e bina y
smoking s a us, as we used, has a low co ela ion o BMI, WC,
and WHR, as es ima ed in he ARIC s udy’s Eu opean descen
pa icipan s ( 0.13, 0.08 and 0.12, espec i ely) and in he
F amingham Hea S udy ( 0.05, 0.08 and 0.16). Addi ionally,
he e a e no loci iden ified in App oach 1 (SNPadjSMK) ha
a e associa ed wi h any smoking beha iou ai and ha exhibi
an opposi e di ec ion o e ec om ha iden ified in ou
adiposi y ai s (Supplemen a y Da a 8). We he e o e p eclude
po en ial collide bias and pos ula e ue gain in powe h ough
SMK-adjus men a hese loci.
To assess how much addi ional in o ma ion is p o ided by
accoun ing o SMK and GxSMK in GWAS o obesi y ai s, we
compa ed gene ic isk sco es (GRSs) based on a ious subse s o
lead SNP geno ypes in a ious eg ession models (Me hods
sec ion). While any GRS was associa ed wi h i s obesi y ai
(P
GRS
o1.6 107, Supplemen a y Table 16), adding SMK and
GxSMK e ms o he eg ession model along wi h no el a ian s
o he GRSs subs an ially inc eased a iance explained. Fo
example, a iance explained inc eased by 38% o BMI ( om
Table 2 | No el loci showing significan associa ion in App oaches 1 (SNPadjSMK) and/o 2 (SNPjoin ) iden ified in seconda y
me a-analyses and no significan in p ima y me a-analyses.
App oach:
S a a
Ma ke Ch :Pos
(hg19)
Nea es
Gene
NEAF Alleles
E/O
Smoke s Non-smoke s Main and in e ac ion e ec s GIANT þUKBB
bP- alue bP- alue b
adj
P
SNPadj
P
SNPin
P
SNPjoin
P
SNPadjSMK
P
SNPin
P
SNPjoin
BMI
1:EC s2481665 1:62594677 INADL 209,453 0.56 T/C 0.015 4.60E 02 0.021 8.90E 08 0.019 3.50E 08 4.00E 01 6.70E 08 3.3E 11 7.8E 01 2.0E 08
1:AW s12629427 3:89145340 EPHA3 137,961 0.26 C/T 0.025 2.10E 02 0.028 3.60E 07 0.027 4.80E 08 8.00E 01 2.00E07 7.7E 08 9.1E01 3.0E 07
1:EW s2173039 3:89142175 117,942 0.26 C/G 0.024 3.10E 02 0.032 8.90E 08 0.031 7.30E 09 5.70E 01 6.50E 08 2.4E 09 9.3E 01 2.2E 07
WCadjBMI
1:EM s1545348 5:34718343 RAI14 77,677 0.73 T/G 0.044 3.10E 04 0.03 1.90E 05 0.034 1.80E 08 3.20E 01 1.70E 07 1.2E 07 1.2E 01 4.8E 07
2:EW s6076699 20:4566688 PRNP 76,930 0.97 A/G 0.169 1.40E 05 0.07 1.20E 04 0.034 3.50E 02 1.40E 08 4.80E 08 4.2E 02 2.3E06 3.4E06
WHRadjBMI
1:AW s670752 3:107312980 BBX 107,568 0.32 A/G 0.012 5.50E 02 0.009 1.50E 02 0.027 4.90E 08 6.80E 01 7.80E 03 3.1E 10 3.8E 01 9.5E 05
1:EC s589428 6:31848220 EHMT2 162,918 0.66 G/T 0.006 1.20E 01 0.011 4.10E 04 0.022 2.80E 08 3.50E 01 7.00E 04 1.1E 17 8.4E 02 1.6E 10
2:EC s1856293 6:133480940 EYA4 127,431 0.52 A/C 0.006 5.30E 01 0.028 9.10E 09 0.019 6.50E 06 5.40E 04 4.70E 08 9.6E 08 1.3E 02 1.5E 08
1:AW s2001945 8:126477978 TRIB1 103,446 0.4 G/C 0.009 1.20E 01 0.013 1.00E 04 0.025 4.70E 08 5.90E 01 1.30E 04 1.1E 09 3.0E 01 1.4E 06
1:EC s17065323 13:44627788 SMIM2* 69,968 0.01 T/C 0.154 1.90E 01 0.23 1.20E 10 0.181 9.20E 09 1.40E 03 3.90E 10 9.6E 09 3.6E 03 1.3E 09
A, all ances ies; C, combined sexes; Ch , ch omosome; E, Eu opean-only; EAF, e ec allele equency; E/O, e ec /o he ; in , in e ac ion; M, men only; Pos, posi ion (bp); Padj, adjus ed o smoking;
W, women only.
All es ima es a e om he s a um specified in he App oach:Sample column.
*This locus was fil e ed om app oaches 2–4 due o low sample size in he SMK s a a, and only P alues o App oach 1 a e conside ed significan . Significan P- alues ha each genome-wide
significance (Po5108) h eshold a e in bold.
Table 3 | Summa y o associa ion esul s o loci showing significance o in e ac ion wi h smoking in App oach (App) 3 (SNPin )
and/o App oach 4 (SNPsc een) in ou p ima y me a-analyses o combined ances ies and combined sexes.
App Ma ke Ch :Pos
(hg19)
Nea es
Gene
NEAF Alleles
E/O
Smoke s Non-smoke s Main and in e ac ion e ec s GIANT þUKBB
bP- alue bP- alue b
adj
P
SNPadj
P
SNPin
P
SNPjoin
P
SNPadjSMK
P
SNPin
P
SNPjoin
BMI
3 s336396 4:143062811 INPP4B 169,646 0.18 T/C 0.063 4.8E08 0.006 3.4E01 0.007 2.3E01 2.1E08 1.9E07 7.4E01 2.7E06 1.3E05
3 s12902602* 15:78967401 CHRNB4 240,135 0.62 A/G 0.047 1.8E 11 0.002 5.5E01 0.009 8.6E03 4.1E11 1.1E10 1.1E01 6.0E13 1.6E12
WCadjBMI
3 s4141488 16:9629067 GRIN2A 153,892 0.5 T/C 0.037 2.2E 05 0.015 9.6E04 0.003 4.4E 01 2.7E 08 5.0E 07 9.5E 01 1.8E 06 1.1E 05
WHRadjBMI
4 s765751* 1:219669226 LYPLAL1 189,028 0.64 C/T 0.003 3.9E 01 0.019 3.1E11 0.029 3.1E16 7.3E 04 2.1E 10 9.1E31 1.4E 04 7.8E22
4 s7766106* 6:127455138 RSPO3 188,174 0.48 T/C 0.007 7.9E02 0.022 2.2E15 0.037 3.7E27 9.7E04 3.8E15 4.4E51 1.0E 05 3.4E 34
Adj, adjus ed o smoking; app, app oach; in , in e ac ion; ch , ch omosome; EAF, e ec allele equency; E/O, e ec /o he ; Pos, posi ion (bp).
*Known locus.
Significan P- alues a e mul iple es co ec ion a e i alicized.
Significan P- alues ha each genome-wide significance (Po5108) h eshold a e in bold.
ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977
8NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions
1.53 o 2.11%, P
GRSDi
¼4.3105), by 27% o WCadjBMI
( om 2.59 o 3.29%, P
GRSDi
¼3.9 106) and by 168% o
WHRadjBMI ( om 0.82 o 2.20%, P
GRSDi
¼3.21011).
The e o e, despi e po en ial limi a ions, much is gained by
accoun ing o en i onmen al exposu es in GWAS s udies.
Discussion
To be e unde s and he e ec s o smoking on gene ic
suscep ibili y o obesi y, we conduc ed me a-analyses o unco e
gene ic a ian s ha may be masked when he en i onmen al
influence o smoking is no conside ed, and o disco e gene ic
loci ha in e ac wi h smoking on adiposi y- ela ed ai s. We
iden ified 161 loci in o al, including 23 no el loci (6 o BMI, 11
o WCadjBMI, and 6 o WHRadjBMI). While many o ou
newly iden ified loci suppo he hypo hesis ha smoking may
influence weigh fluc ua ions h ough appe i e egula ion, hese
no el loci also ha e highligh ed new biological p ocesses and
pa hways implica ed in he pa hogenesis o obesi y.
Impo an ly, we iden ified nine loci wi h con incing e idence o
GxSMK in e ac ion on obesi y- ela ed ai s. We we e able o
eplica e he p e ious GxSMK in e ac ion wi h BMI wi hin he
CHRNA5-CHRNA3-CHRNB4 gene clus e . One no el BMI-
associa ed locus nea INPP4B and wo no el WCadjBMI-associa ed
loci nea GRIN2A and PRNP displayed significan GxSMK
in e ac ion. We we e also able o iden i y significan GxSMK
in e ac ion o one known BMI-associa ed locus nea ADAMTS7
and o fi e known WHRadjBMI-associa ed loci nea LYPLAL1,
RSPO3,MAP3K1,HOXC4-HOXC6 and JUND. The majo i y o
hese loci ha bou s ong candida e genes o adiposi y wi h a
possible ole o he modula ion o e ec s h ough obacco use.
We iden ified 18 new loci in App oach 1 (P
SNPadjSMK
)by
adjus ing o cu en smoking s a us. Ou analyses did no allow
us o de e mine whe he hese disco e ies a e due o di e en
subse s o subjec s included in he analyses compa ed o p e ious
s udies1,2 o due only o adjus ing o cu en smoking.
Adjus men o cu en smoking in ou analyses, howe e , did
e eal no el associa ions. Specifically a e accoun ing o
smoking in ou analyses, all no el BMI loci exhibi P- alues
ha a e a leas one o de o magni ude lowe han in
p e ious GIANT in es iga ions, despi e smalle samples in he
cu en analysis2. While sample sizes o bo h WCadjBMI
and WHRadjBMI a e compa able wi h p e ious GIANT
in es iga ions, ou P alues o a ian s iden ified in App oach
1 a e a leas wo o de s o magni ude lowe han p e ious
findings. Thus, adjus men o smoking may ha e indeed e ealed
new loci. Fu he , loci iden ified in App oach 2, including nine
no el loci, sugges ha accoun ing o in e ac ion imp o es ou
abili y o de ec hese loci e en in he p esence o only modes
e idence o GxSMK in e ac ion.
The e a e se e al challenges in alida ing gene ic associa ions
ha accoun o en i onmen al exposu e. In addi ion o exposu e
ha moniza ion and po en ial bias due o adjus men o smoking
exposu e, di e ences in ai dis ibu ion, en i onmen al expo-
su e equency, ances y-specific LD pa e ns and allele equency
ac oss s udies may lead o di ficul ies in eplica ion, especially o
gene-by-en i onmen s udies54. Fu he mo e, he ‘winne ’s cu se’
(infla ed disco e y e ec s es ima es) equi es la ge sample
sizes o adequa e powe in eplica ion s udies55. Despi e hese
challenges, we we e able o de ec consis en di ec ion o e ec in
an independen sample o all no el loci. Some esul s ha did
no emain GWS in he GIANT þUKBB me a-analysis had
esul s ha we e jus unde he h eshold o significance,
sugges ing ha a la ge sample may be needed o confi m hese
esul s, and hus he associa ions nea INPP4B,GRIN2A,RAI14,
PRNP and JUND should be in e p e ed wi h cau ion.
While we ound ha e ec s we e no significan ly en iched in
smoke s o BMI, he e is a g ea e p opo ion o a iance in BMI
explained by a ian s ha a e significan o App oach 1
(SNPadjSMK), which may be expec ed gi en ha he e a e a
g ea e numbe o a ian s wi h highe e ec es ima es in
smoke s. Fo WCadjBMI, he e was no en ichmen o s onge
e ec s in one s a um compa ed o he o he o ou significan
loci; howe e , he e was a g ea e p opo ion o explained
a iance in WCadjBMI o loci iden ified in App oach 1
(SNPadjSMK) in nonsmoke s. Fo WHRadjBMI, he e we e
significan ly mo e loci ha exhibi g ea e e ec s in nonsmoke s,
and his pa e n was mi o ed in he a iance explained analysis.
The la ge di e ence be ween e ec s in smoke s and nonsmoke s
likely explains he sub-GWS le els o ou loci in p e ious GIANT
in es iga ions2. Fo example, he T allele o s7697556, 81kb om
he ADAMTS3 gene, was associa ed wi h inc eased WCadjBMI
and exhibi s a six old g ea e e ec in nonsmoke s compa ed o
smoke s, al hough he in e ac ion e ec was only nominal; in
p e ious GWAS his a ian was nea ly GWS. These di e ences
in e ec es ima es be ween smoke s and nonsmoke s may help
explain inconsis en findings in p e ious analyses ha show
cen al adiposi y inc eases wi h inc eased smoking, bu is
associa ed wi h dec eased weigh and BMI5,9,10.
Ou esul s suppo p e ious findings ha implica e genes
in ol ed in ansc ip ion and gene exp ession, appe i e egula-
ion, mac onu ien me abolism, and glucose homeos asis. Se e al
o ou no el loci ha e candida e genes wi hin 500 kb o ou ag
Table 4 | Summa y o associa ion esul s o loci showing significance o in e ac ion wi h smoking in App oach 3 (SNPin )
and/o App oach 4 (SNPsc een) in ou seconda y me a-analyses no iden ified in p ima y me a-analyses.
App oach:
S a a
Ma ke Ch :Pos
(hg19)
Nea es
Gene
NEAF Alleles
E/O
Smoke s Non-smoke s Main and in e ac ion e ec s GIANT þUKBB
bPbPb
adj
P
SNPadj
P
SNPin
P
SNPjoin
P
SNPadjSMK
P
SNPin
P
SNPjoin
BMI
4:AM s1809420* 15:79056769 ADAMTS7 57,081 0.59 T/C 0.074 9.8E 08 0.023 2.0E 03 0.036 4.9E 08 9.4E 04 5.6E09 9.8E 05 3.3E 05 1.9E 07
WCadjBMI
3:EW s6076699 20:4566688 PRNP 76,930 0.97 A/G 0.169 1.4E 05 0.07 1.2E 04 0.034 3.5E 02 1.4E 08 4.8E 08 4.2E02 2.3E 06 3.4E06
WHRadjBMI
4:EM s30000* 5:55803533 MAP3K1 71,424 0.27 G/A 0.002 7.8E 01 0.031 3.7E 08 0.04 1.7E 10 1.6E04 2.7E 07 2.7E 17 3.2E 07 3.8E 15
4:AM s459193* 5:55806751 80,852 0.27 A/G 0.004 5.0E 01 0.034 4.1E10 0.043 2.3E 13 6.8E 05 2.2E 09 3.5E 20 2.5E07 1.6E 17
4:AM s2071449* 12:54428011 HOXC4- 70,868 0.37 A/C 0.003 6.0E 01 0.026 1.0E06 0.034 9.1E 09 1.1E03 5.7E 06 2.7E 12 8.0E 04 2.8E 09
4:EM s754133* 12:54418920 HOXC6 71,136 0.36 A/G 0.003 6.2E 01 0.026 8.2E 07 0.034 3.0E 09 1.1E 03 4.0E 06 2.1E 12 9.7E 04 4.0E09
4:AM s12608504* 19:18389135 JUND 80,087 0.37 A/G 0.006 2.6E01 0.025 5.0E 07 0.032 4.7E09 5.5E 03 1.8E 06 2.9E 11 1.3E 02 1.6E 08
A, all ances ies; Adj, adjus ed o smoking; app, app oach; C, combined sexes; Ch , ch omosome; E, Eu opean-only; EAF, e ec allele equency; E/O, e ec /o he ; in , in e ac ion; M, men only;
Pos, posi ion (bp); W, women only.
All es ima es a e om he s a um specified in he App oach:Sample column The R2be ween he ADAMTS7 ( s1809420) and CHRNB4 a ian ( s1290362) in Table 3 is 0.72 (HapMap 2, CEU).
Addi ionally, he PRNP a ian ( s6076699) is he same as he a ian ha came up om App oach 2 (Table 2).
*Known locus.
Significan P- alues a e mul iple es co ec ion a e i alicized.
Significan P- alues ha each genome-wide significance (Po5108) h eshold a e in bold.
NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977 ARTICLE
NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions 9
Jaakko Tuomileh o187,188,190, And e G. Ui e linden32,191, Liesbe h Vandenpu 34, Ma ie-Claude Vohl58,192,
Hen y Vo
¨lzke68, Judi h M. Vonk72,Ge
´ a d Waebe 158, Melanie Waldenbe ge 108,109, R.G.J. Wes endo p193,
Sa ah Wild98, Gonneke Willemsen77, B uce H.R. Wol enbu el73, And ew Wong194, Alan F. W igh 39,
Wei Zhao25, M. Ca ola Zillikens191, Damiano Baldassa e195,196, Be e ley Balkau197, S e ania Bandinelli198,
Ca s en A. Bo
¨ge 36, Do e I. Boomsma77, Claude Boucha d170, Ma cel B uinenbe g199, Daniel I. Chasman11,200,
Yii-De Ida Chen201, Pe e S. Chines93, Richa d S. Coope 186, F ancesco Cucca174,202, Daniele Cusi203,
Ul de Fai e106, Luigi Fe ucci67, Paul W. F anks19,21,204, Philippe F oguel26,205, Penny Go don-La sen83,206,
Hans-Jo
¨ gen G abe207,208, Vilmundu Gudnason61,62, Ch is ophe A. Haiman209, Ca oline Haywa d39,118,
K is ian H eem145, And ew D. Johnson13, J. Wou e Jukema69,210,211, Sha on L.R. Ka dia25, Mika Ki imaki45,
Jaspal S. Koone 81,99,212, Diana Kuh194, Ma kku Laakso144, Te ho Leh ima
¨ki48,49, Loic Le Ma chand46,
Win ied Ma
¨ z213,214, Ma k I. McCa hy37,17,215, And es Me spalu35, And ew P. Mo is17,216, Claes Ohlsson34,
Lyle J. Palme 217, Ge a d Pas e kamp71,218, Olu Pede sen8, Anne e Pe e s109,110, Ul ike Pe e s82, Oz en
Polasek98,142, B uce M. Psa y219,220,221,LuQi
21,222, Raine Rau amaa43,223, Blai H. Smi h118,224, Tho kild I.A.
Sø ensen8,225,226, Kons an in S auch54,157, Henning Tiemeie 227, Elena T emoli195,196, Pim an de
Ha s 76,183,228, Hen ik Ves e gaa d8,9, Pe e Vollenweide 158, Nicholas J. Wa eham47, Da id R. Wei 103,
John B. Whi field53, James F. Wilson39,229, Jessica Ty ell230,231, Timo hy M. F ayling232, Ine
ˆs Ba oso233,234,235,
Michael Boehnke28, Panagio is Deloukas40,233,236, Ca oline S. Fox10, Joel N. Hi schho n74,75,237,
Da id J. Hun e 21,75,238,239, Tim D. Spec o 50, Da id P. S achan5,240, Co nelia M. an Duijn24,241,242,
I is M. Heid2,243, Ka en L. Mohlke79, Jona han Ma chini244, Ru h J.F. Loos22,23,47,245,246,**,
Tuomas O. Kilpela
¨inen8,47,247,**, Ching-Ti Liu4,**, Ing id B. Bo ecki3,**, Ka i E. No h1,**
& L. Ad ienne Cupples4,10,**
1Depa men o Epidemiology, Uni e si y o No h Ca olina, Chapel Hill, No h Ca olina 27599, USA. 2Depa men o Gene ic Epidemiology, Ins i u e o
Epidemiology and P e en i e Medicine, Uni e si y o Regensbu g, D-93053 Regensbu g, Ge many. 3Di ision o S a is ical Genomics, Depa men o
Gene ics, Washing on Uni e si y School o Medicine; S . Louis, Missou i 63108, USA. 4Depa men o Bios a is ics, Bos on Uni e si y School o Public Heal h,
Bos on, Massachuse s 02118, USA. 5Popula ion Heal h Resea ch Ins i u e, S . Geo ge’s, Uni e si y o London, London SW17 0RE, UK. 6T ansMed Sys ems,
Inc., Cupe ino, Cali o nia 95014, USA. 7Depa men o Bios a is ics, Johns Hopkins Bloombe g School o Public Heal h, Bal imo e, Ma yland, USA.
8The No o No disk Founda ion Cen e o Basic Me abolic Resea ch, Sec ion o Me abolic Gene ics, Facul y o Heal h and Medical Sciences, Uni e si y o
Copenhagen, Copenhagen, Denma k. 9S eno Diabe es Cen e , Gen o e, Denma k. 10 NHLBI F amingham Hea S udy, F amingham, Massachuse s 01702,
USA. 11 Di ision o P e en i e Medicine, B igham and Women’s Hospi al and Ha a d Medical School, Bos on, Massachuse s, USA. 12 Depa men o
Neu ology, Bos on Uni e si y School o Medicine, Bos on, Massachuse s 02118, USA. 13 Popula ion Sciences B anch, Na ional Hea , Lung, and Blood
Ins i u e, Na ional Ins i u es o Heal h, The F amingham Hea S udy, F amingham, Massachuse s, USA. 14 Ins i u e o Social and P e en i e Medicine
(IUMSP), Cen e Hospi alie Uni e si ai e Vaudois (CHUV), Lausanne, Swi ze land. 15 Depa men o Compu a ional Biology, Uni e si y o Lausanne,
Lausanne, Swi ze land. 16 Swiss ins i i u e o Bioin o ma ics, 1015 Lausanne, Swi ze land. 17 Wellcome T us Cen e o Human Gene ics, Uni e si y o Ox o d,
Ox o d OX3 7BN, UK. 18 Depa men o Biobank Resea ch, Umeå Uni e si y, Umeå, Sweden. 19 Depa men o Clinical Sciences, Gene ic and Molecula
Epidemiology Uni , Lund Uni e si y, SE-205 02 Malmo¨, Sweden. 20 Depa men o Epidemiology and Popula ion Heal h, Albe Eins ein College o Medicine,
B onx, New Yo k, USA. 21 Depa men o Nu i ion, Ha a d T.H. Chan School o Public Heal h, Bos on, Massachuse s 02115, USA. 22 The Cha les B on man
Ins i u e o Pe sonalized Medicine, Icahn School o Medicine a Moun Sinai, New Yo k, USA. 23 The Gene ics o Obesi y and Rela ed Me abolic T ai s
P og am, Icahn School o Medicine a Moun Sinai, New Yo k, USA. 24 Gene ic Epidemiology Uni , Depa men o Epidemiology, E asmus Uni e si y Medical
Cen e , Ro e dam 3015GE, The Ne he lands. 25 Depa men o Epidemiology, School o Public Heal h, Uni e si y o Michigan, Ann A bo , Michigan, USA.
26 Uni e si y o Lille, CNRS, Ins i u Pas eu o Lille, UMR 8199 - EGID, Lille, F ance. 27 In e nal Medicine - Neph ology, Uni e si y o Michigan, Ann A bo ,
Michigan, USA. 28 Depa men o Bios a is ics and Cen e o S a is ical Gene ics, Uni e si y o Michigan, Ann A bo , Michigan 48109, USA. 29 Cen e o
Gene ic O igins o Heal h and Disease, Uni e si y o Wes e n Aus alia, C awley 6009, Aus alia. 30 Depa men o Heal h Sciences, Uni e si y o Milan,Via
A. Di Rudinı
´, 8 20142, Milano, I aly. 31 Depa men o Bios a is ics, Uni e si y o Washing on, Sea le, Washing on 98195, USA. 32 Depa men o
Epidemiology, E asmus Medical Cen e , Ro e dam, The Ne he lands. 33 Depa men o Psychia y, Dokuz Eylul Uni e si y, Izmi , Tu key. 34 Cen e o Bone
and A h i is Resea ch, Depa men o In e nal Medicine and Clinical Nu i ion, Ins i u e o Medicine, Sahlg enska Academy a he Uni e si y o Go henbu g,
Go henbu g, Sweden. 35 Es onian Genome Cen e , Uni e si y o Ta u, Ta u 51010, Es onia. 36 Depa men o Neph ology, Uni e si y Hospi al Regensbu g,
Regensbu g, Ge many. 37 Ox o d Cen e o Diabe es, Endoc inology and Me abolism, Uni e si y o Ox o d, Chu chill Hospi al, Ox o d OX3 7LJ, UK.
38 Epidemiology Domain, Saw Swee Hock School o Public Heal h, Na ional Uni e si y o Singapo e, Singapo e 117549, Singapo e. 39 MRC Human Gene ics
Uni , Ins i u e o Gene ics and Molecula Medicine, Uni e si y o Edinbu gh, Edinbu gh, Sco land. 40 William Ha ey Resea ch Ins i u e, Ba s and The London
School o Medicine and Den is y, Queen Ma y Uni e si y o London, London, UK. 41 Depa men o Heal h, Na ional Ins i u e o Heal h and Wel a e, Helsinki
FI-00271, Finland. 42 V h Depa men o Medicine, Medical Facul y Mannheim, Heidelbe g Uni e si y, Mannheim, Ge many. 43 Kuopio Resea ch Ins i u e o
Exe cise Medicine, Kuopio, Finland. 44 ISER, Uni e si y o Essex, Colches e CO43SQ, UK. 45 Depa men o Epidemiology and Public Heal h, UCL, London,
WC1E 6BT, UK. 46 Epidemiology P og am, Uni e si y o Hawaii Cance Cen e , Honolulu, Hawaii 96813, USA. 47 MRC Epidemiology Uni , Uni e si y o
Camb idge School o Clinical Medicine, Ins i u e o Me abolic Science, Camb idge CB2 0QQ, UK. 48 Depa men o Clinical Chemis y, Fimlab Labo a o ies,
Tampe e 33520, Finland. 49 Depa men o Clinical Chemis y, Facul y o Medicine and Li e Sciences, Uni e si y o Tampe e, Tampe e 33014, Finland.
ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977
16 NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions
50 Depa men o Twin Resea ch and Gene ic Epidemiology, King’s College London, London, UK. 51 NIHR Biomedical Resea ch Cen e a Guy’s and S .
Thomas’ Founda ion T us , London, UK. 52 Cen e o Public Heal h Genomics and Bios a is ics Sec ion, Depa men o Public Heal h Sciences, Uni e si y o
Vi ginia, Cha lo es ille, Vi ginia 22903, USA. 53 Gene ic Epidemiology, QIMR Be gho e Medical Resea ch Ins i u e, B isbane 4029 , Aus alia. 54 Ins i u e o
Gene ic Epidemiology, Helmhol z Zen um Mu
¨nchen - Ge man Resea ch Cen e o En i onmen al Heal h, D-85764 Neuhe be g, Ge many. 55 Depa men o
Medicine I, Uni e si y Hospi al G osshade n, Ludwig-Maximilians-Uni e si a
¨ , D-81377 Munich, Ge many. 56 DZHK (Ge man Cen e o Ca dio ascula
Resea ch), pa ne si e Munich Hea Alliance, Munich, Ge many. 57 Depa men o Kinesiology, Facul y o Medicine, Uni e si e
´La al, Quebec Ci y, Que
´bec,
Canada, G1V 0A6. 58 Ins i u e o Nu i ion and Func ional Foods, Uni e si e
´La al, Quebec Ci y, Que
´bec, Canada, G1V 0A6. 59 Depa men o Bio echnology,
Ins i u e o Molecula and Cell Biology, Uni e si y o Ta u, Ta u 51010, Es onia. 60 Depa men o Social and Heal h Ca e, Ci y o Helsinki, Helsinki, Finland.
61 Icelandic Hea Associa ion, Kopa ogu , Iceland. 62 Facul y o Medicine, Uni e si y o Iceland, Reykja ik, Iceland. 63 Depa men o Medicine, Ins i u e o
Clinical Medicine, Uni e si y o Eas e n Finland, 70210 Kuopio, Finland.. 64 Ca dio ascula Medicine Uni , Depa men o Medicine Solna, Ka olinska Ins i u e ,
S ockholm, Sweden. 65 Cen e o Molecula Medicine, Ka olinska Uni e si y Hospi al Solna, S ockholm, Sweden. 66 Di ision o Bios a is ics, Washing on
Uni e si y School o Medicine, S Louis, Missou i, USA. 67 T ansla ional Ge on ology B anch, Na ional Ins i u e on Aging, Bal imo e, Ma yland, USA.
68 Ins i u e o Communi y Medicine, Uni e si y Medicine G ei swald, Ge many. 69 Depa men o Ca diology, Leiden Uni e si y Medical Cen e , Leiden, The
Ne he lands. 70 Depa men o Ge on ology and Ge ia ics, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands. 71 Labo a o y o Expe imen al
Ca diology, Depa men o Ca diology, Di ision Hea & Lungs, UMC U ech , U ech , The Ne he lands. 72 Depa men o Epidemiology, Uni e si y o
G oningen, Uni e si y Medical Cen e G oningen, The Ne he lands. 73 Depa men o Endoc inology, Uni e si y o G oningen, Uni e si y Medical Cen e
G oningen, G oningen, The Ne he lands. 74 Di isions o Endoc inology and Gene ics and Cen e o Basic and T ansla ional Obesi y Resea ch, Bos on
Child en’s Hospi al, Bos on, Massachuse s 02115, USA. 75 B oad Ins i u e o Ha a d and MIT, Camb idge, Massachuse s 02142, USA. 76 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. 77 Depa men o Biological Psychology, V ije
Uni e si ei , Ams e dam, The Ne he lands. 78 Beha iou al Science Ins i u e, Radboud Uni e si y, Nijmegen, The Ne he lands. 79 Depa men o Gene ics,
Uni e si y o No h Ca olina, Chapel Hill, No h Ca olina 27599, USA. 80 Dep Epidemiology and Bios a is ics, School o Public Heal h, Impe ical College
London, UK. 81 Ca diology, Ealing Hospi al NHS T us , Middlesex, UK. 82 Di ision o Public Heal h Sciences, F ed Hu chinson Cance Resea ch Cen e , Sea le,
Washing on, USA. 83 Depa men o Nu i ion, Gillings School o Global Public Heal h, Uni e si y o No h Ca olina a Chapel Hill, Chapel Hill, No h Ca olina
27599, USA. 84 Depa men o Medicine, Uni e si y Medical Cen e G oningen, Uni e si y o G oningen, G oningen, The Ne he lands. 85 Ca dio ascula
Heal h Resea ch Uni , Depa men o Medicine, Uni e si y o Washing on, Sea le, Washing on 98101, USA. 86 Bussel on Popula ion Medical Resea ch
Ins i u e, Nedlands, Wes e n Aus alia 6009, Aus alia. 87 Pa hWes Labo a o y Medicine o WA, Si Cha les Gai dne Hospi al, Nedlands, Wes e n Aus alia
6009, Aus alia. 88 School o Pa hology and Labo a y Medicine, The Uni e si y o Wes e n Aus alia, 35 S i ling Hwy, C awley, Wes e n Aus alia 6009,
Aus alia. 89 Diabe es and Obesi y Resea ch Ins i u e, Ceda s-Sinai Medical Cen e , Los Angeles, Cali o nia, USA. 90 Clinic o P os he ic Den is y,
Ge os oma ology and Ma e ial Science, Uni e si y Medicine G ei swald, G ei swald, Ge many. 91 Sou h Texas Diabe es and Obesi y Ins i u e, Uni e si y o
Texas Rio G ande Valley, B owns ille, Texas, USA. 92 Human Gene ics Cen e , The Uni e si y o Texas Heal h Science Cen e , PO Box 20186, Hous on, Texas
77225, USA. 93 Medical Genomics and Me abolic Gene ics B anch, Na ional Human Genome Resea ch Ins i u e, NIH, Be hesda, Ma yland 20892, USA.
94 Depa men o Pha macology and Sys ems The apeu ics, Icahn School o Medicine a Moun Sinai, New Yo k, USA. 95 Depa men o Pha macology and
The apeu ics, Uni e si y College Co k, Co k, I eland. 96 Depa men o Gene ics, Ru ge s Uni e si y, Pisca away, New Je sey 08854, USA. 97 Depa men o
S a is ics and Bios a is ics, Ru ge s Uni e si y, Pisca away, New Je sey 08854, USA. 98 Ushe Ins i u e o Popula ion Heal h Sciences and In o ma ics, The
Uni e si y o Edinbu gh, Sco land, UK. 99 Impe ial College Heal hca e NHS T us , London, UK. 100 Depa men o Vascula Su ge y, Di ision o Su gical
Special ies, UMC U ech , U ech , The Ne he lands. 101 EMGO þIns i u e V ije Uni e si ei & V ije Uni e si ei Medical Cen e , Ams e dam, he Ne he lands.
102 Depa men o Nu i ion and Die e ics, School o Heal h Science and Educa ion, Ha okopio Uni e si y, A hens, G eece. 103 Su ey Resea ch Cen e ,
Ins i u e o Social Resea ch, Uni e si y o Michigan, Ann A bo , Michigan, USA. 104 Robe son Cen e o Bios a is ics, Uni e si y o Glasgow, Glasgow, UK.
105 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 JMAAW15, Jamaica. 106 Uni o
Ca dio ascula Epidemiology, Ins i u e o En i onmen al Medicine, Ka olinska Ins i u e , S ockholm, Sweden. 107 Hype ension and Rela ed Disease Cen e,
AOU-Uni e si y o Sassa i, Sassa i, I aly. 108 Resea ch Uni o Molecula Epidemiology, Helmhol z Zen um Mu
¨nchen, Ge man Resea ch Cen e o
En i onmen al Heal h, D-85764 Neuhe be g, Ge many. 109 Ins i u e o Epidemiology II, Helmhol z Zen um Mu
¨nchen - Ge man Resea ch Cen e o
En i onmen al Heal h, D-85764 Neuhe be g, Ge many. 110 Ge man Cen e o Diabe es Resea ch, D-85764 Neuhe be g, Ge many. 111 Depa men o Public
Heal h and Clinical Medicine, Sec ion o Nu i ional Resea ch, Umeå Uni e si y, Umeå, Sweden. 112 Labo a o y o Epidemiology, Demog aphy, and Biome y,
Na ional Ins i u e on Aging, Na ional Ins i u es o Heal h, Be hesda, Ma yland, USA. 113 In e disciplina y Cen e Psychopa hology and Emo ion Regula ion
(ICPE), Uni e si y o G oningen, Uni e si y Medical Cen e G oningen, G oningen, The Ne he lands. 114 Depa men o Psychia y, Washing on Uni e si y
School o Medicine, S . Louis, Missou i, USA. 115 Labo a o y o Neu ogene ics, Na ional Ins i u e on Aging, Be hesda, Ma yland, USA. 116 Di ision o Genomic
Medicine, Na ional Human Genome Resea ch Ins i u e, Na ional Ins i u es o Heal h, Be hesda, Ma yland 20892, USA. 117 Ins i u e o Medical Sciences,
Uni e si y o Abe deen, Fo es e hill, Abe deen AB25 2ZD, UK. 118 Gene a ion Sco land, Cen e o Genomic and Expe imen al Medicine, Uni e si y o
Edinbu gh, Edinbu gh, Sco land. 119 S . Ola Hospi al, T ondheim Uni e si y Hospi al, T ondheim, No way. 120 In e acul y Ins i u e o Gene ics and Func ional
Genomics, Uni e si y Medicine G ei swald, G ei swald, Ge many. 121 Depa men o Human Gene ics, Wellcome T us Sange Ins i u e, Hinx on, Camb idge,
UK. 122 School o Medicine and Pha macology, The Uni e si y o Wes e n Aus alia, 25 S i ling Hwy, C awley, Wes e n Aus alia 6009, Aus alia.
123 Depa men o Ca dio ascula Medicine, Si Cha les Gai dne Hospi al, Nedlands, Wes e n Aus alia 6009, Aus alia. 124 Depa men o Pedia ics,
Tampe e Uni e si y Hospi al, Tampe e 33521, Finland. 125 Depa men o Pedia ics, Facul y o Medicine and Li e Sciences, Uni e si y o Tampe e, Tampe e
33014, Finland. 126 Depa men o Medical Sciences, Molecula Epidemiology, Uppsala Uni e si y, Uppsala, 751 85, Sweden. 127 Depa men o Medicine,
Di ision o Ca dio ascula Medicine, S an o d Uni e si y School o Medicine, S an o d, Cali o nia 94305, USA. 128 Science o Li e Labo a o y, Uppsala
Uni e si y, Uppsala 750 85, Sweden. 129 Depa men o Pulmona y Physiology and Sleep Medicine, Si Cha les Gai dne Hospi al, Nedlands, Wes e n
Aus alia 6009, Aus alia. 130 Depa men o Physiology, Ins i u e o Neu oscience and Physiology, he Sahlg enska Academy a he Uni e si y o Go henbu g,
Go henbu g, Sweden. 131 Depa men o Epidemiology and Bios a is ics, MRC–PHE Cen e o En i onmen & Heal h, School o Public Heal h, Impe ial College
London, No olk Place, London, UK. 132 Cen e o Li e Cou se Epidemiology, Facul y o Medicine, Uni e si y o Oulu, P.O.Box 5000, FI-90014, Oulu, Finland.
133 Biocen e Oulu, Uni e si y o Oulu, Oulu, Finland. 134 Uni o P ima y Ca e, Oulu Uni e si y Hospi al, Kajaanin ie 50, P.O.Box 20, FI-90220, 90029 Oulu,
Finland. 135 Depa men o Medicine, Uni e si y o Tu ku, Tu ku 20520, Finland. 136 Di ision o Medicine, Tu ku Uni e si y Hospi al, Tu ku 20521, Finland.
137 Depa men o Clinical Physiology, Tampe e Uni e si y Hospi al, Tampe e 33521, Finland. 138 Depa men o Clinical Physiology, Facul y o Medicine and
Li e Sciences, Uni e si y o Tampe e, Tampe e 33014, Finland. 139 Clinical and Molecula Os eopo osis Resea ch Uni , Depa men o O hopedics and Clinical
Sciences, Skåne Uni e si y Hospi al, Lund Uni e si y, Malmo¨, Sweden. 140 Depa men o Medicine and Abdominal Cen e : Endoc inology, Uni e si y o
Helsinki and Helsinki Uni e si y Cen al Hospi al, Helsinki FI-00029, Finland. 141 Mine a Founda ion Ins i u e o Medical Resea ch, Biomedicum 2U, Helsinki
FI-00290, Finland. 142 Depa men o Public Heal h, Facul y o Medicine, Uni e si y o Spli , Spli , C oa ia. 143 Depa men o Ca diology, Onassis Ca diac
Su ge y Cen e , A hens, G eece. 144 Depa men o Medicine, Uni e si y o Eas e n Finland and Kuopio Uni e si y Hospi al, 70210 Kuopio, Kuopio, Finland.
145 HUNT Resea ch Cen e, Depa men o Public Heal h and Nu sing, No wegian Uni e si y o Science and Technology, 7600 Le ange , No way. 146 Ins i u e
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NATURE COMMUNICATIONS | 8:14977 | DOI: 10.1038/ncomms14977 | www.na u e.com/na u ecommunica ions 17
o Biomedicine/Physiology, Uni e si y o Eas e n Finland, Kuopio Campus, Finland. 147 USC-O fice o Popula ion S udies Founda ion, Inc., Uni e si y o San
Ca los, Cebu Ci y 6000, Philippines. 148 Depa men o An h opology, Sociology and His o y, Uni e si y o San Ca los, Cebu Ci y 6000, Philippines.
149 Depa men o Medical Sciences, Ca dio ascula Epidemiology, Uppsala Uni e si y, Uppsala 751 85, Sweden. 150 Li Ka Shing Cen e o Heal h In o ma ion
and Disco e y, The Big Da a Ins i u e, Uni e si y o Ox o d, Ox o d OX3 7BN, UK. 151 Resea ch Cen e o P e en ion and Heal h, he Capi al Region o
Denma k, Copenhagen, Denma k. 152 Depa men o Clinical Expe imen al Resea ch, Rigshospi ale , Glos up, Denma k. 153 Depa men o Clinical Medicine,
Facul y o Heal h and Medical Sciences, Uni e si y o Copenhagen, Copenhagen, Denma k. 154 T ansla ional Labo a o y in Gene ic Medicine (TLGM), Agency
o Science, Technology and Resea ch (A*STAR), 8A Biomedical G o e, Immunos, Le el 5, Singapo e 138648, Singapo e. 155 Depa men o Public Heal h and
P ima y Ca e, Uni e si y o Camb idge, Camb idge, UK. 156 Depa men o Psychology, Uni e si y o No e Dame, No e Dame, USA. 157 Ins i u e o Medical
In o ma ics, Biome y and Epidemiology, Chai o Gene ic Epidemiology, Ludwig-Maximilians-Uni e si a
¨ , D-81377 Munich, Ge many. 158 Depa men o
Medicine, In e nal Medicine, Lausanne uni e si y hospi al (CHUV), Lausanne, Swi ze land. 159 P og am in Bios a is ics and Bioma hema ics, F ed Hu chinson
Cance Resea ch Cen e , Sea le, Washing on 98109, USA. 160 Molecula Epidemiology, QIMR Be gho e Medical Resea ch Ins i u e, B isbane, Queensland
4029, Aus alia. 161 School o Popula ion Heal h, The Uni e si y o Wes e n Aus alia, 35 S i ling Hwy, C awley, Wes e n Aus alia 6009, Aus alia.
162 Depa men o Respi a o y Medicine, Si Cha les Gai dne Hospi al, Nedlands, Wes e n Aus alia 6009, Aus alia. 163 Ins i u e o Clinical Chemis y and
Labo a o y Medicine, Uni e si y Medicine G ei swald, G ei swald, Ge many. 164 Ins i u e o Ca dio ascula and Medical Sciences, BHF Glasgow
Ca dio ascula Resea ch Cen e, Uni e si y o Glasgow, Glasgow, Sco land. 165 Resea ch Cen e o P e en ion and Heal h, Glos up Hospi al, Glos up,
Denma k. 166 Depa men o Public Heal h, Facul y o Heal h Sciences, Uni e si y o Copenhagen, Copenhagen, Denma k. 167 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, Sco land. 168 Depa men o Clinical Physiology and
Nuclea Medicine, Tu ku Uni e si y Hospi al, Tu ku 20521, Finland. 169 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 20520, Finland. 170 Human Genomics Labo a o y, Penning on Biomedical Resea ch Cen e , Ba on Rouge, Louisiana, USA. 171 Depa men o
Gene ics, Washing on Uni e si y School o Medicine, S . Louis, Missou i, USA. 172 Depa men o P e en i e Medicine, No hwes e n Uni e si y Feinbe g
School o Medicine, Chicago, Illinois, USA. 173 Di ision o Ca diology, B igham and Women’s Hospi al, Bos on, Massachuse s, USA. 174 Is i u o di Rice ca
Gene ica e Biomedica (IRGB), Consiglio Nazionale Delle Rice che (CNR), Ci adella Uni e si a ia di Monse a o, 09042 Monse a o, I aly. 175 BHF Glasgow
Ca dio ascula Resea ch Cen e, Facul y o Medicine, Glasgow, UK. 176 Labo a o y o Gene ics, Na ional Ins i u e on Aging, Na ional Ins i u es o Heal h,
Bal imo e, Ma yland, USA. 177 Science o Li e Labo a o y, Ka olinska Ins i u e , S ockholm, Sweden. 178 Di ision o Angiology, Depa men o In e nal
Medicine, Medical Uni e si y o G az, G az, Aus ia. 179 Depa men o Molecula Epidemiology, Leiden Uni e si y Medical Cen e , Leiden, The Ne he lands.
180 Resea ch Uni Hype ension and Ca dio ascula Epidemiology, Depa men o Ca dio ascula Science , Uni e si y o Leu en, Campus Sin Ra ael,
Kapucijnen oe 35, Leu en, Belgium. 181 R&D Vi aK G oup, Maas ich Uni e si y, B ains Unlimi ed Building, Ox o dlaan 55, Maas ich , The Ne he lands.
182 Ins i u e o Ca dio ascula and Medical Sciences, Facul y o Medicine, Uni e si y o Glasgow, Glasgow, UK. 183 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. 184 Cen 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, Cali o nia, USA. 185 Depa men o Pedia ics, Uni e si y o Cali o nia Los Angeles,
Los Angeles, Cali o nia, USA. 186 Depa men o Public Heal h Sciences, S i ch School o Medicine, Loyola Uni e si y o Chicago, Maywood, Illinois 61053,
USA. 187 Resea ch Di ision, Dasman Diabe es Ins i u e, Dasman, Kuwai . 188 Depa men o Neu osciences and P e en i e Medicine, Danube-Uni e si y
K ems, 3500 K ems, Aus ia. 189 Ch onic Disease P e en ion Uni , Na ional Ins i u e o Heal h and Wel a e, Helsinki, Finland. 190 Saudi Diabe es Resea ch
G oup, King Abdulaziz Uni e si y, Jeddah, Saudi A abia. 191 Depa men o In e nal Medicine, E asmus Medical Cen e , Ro e dam, The Ne he lands.
192 School o Nu i ion, Uni e si e
´La al, La al, Que
´bec, Canada. 193 Depa men o Public Heal h and Cen e o Heal hy Aging, Uni e si y o Copenhagen,
1014 Copenhagen, Denma k. 194 MRC Uni o Li elong Heal h and Ageing a UCL, 33 Bed o d Place, London WC1B 5JU, UK. 195 Dipa imen o di Scienze
Fa macologiche e Biomolecola i, Uni e si a
`di Milano, Milan, I aly. 196 Cen o Ca diologico Monzino, IRCCS, Milan, I aly. 197 Inse m U-1018, CESP, 94807
Villejui cedex, F ance. 198 Ge ia ic Uni , Azienda USL Toscana cen o, Flo ence, I aly. 199 Li elines Coho S udy, PO Box 30001, 9700 RB G oningen, The
Ne he lands. 200 Di ision o Gene ics, B igham and Women’s Hospi al, Bos on, Massachuse s, USA. 201 Ins i u e o T ansla ional Genomics and Popula ion
Sciences, Los Angeles BioMedical Resea ch Ins i u e and Depa men o Pedia ics, Ha bo -UCLA, To ance, Cali o nia 90502, USA. 202 Dipa imen o di
Scienze Biomediche, Uni e si a’ degli S udi di Sassa i, Sassa i, I aly. 203 Sanipedia s l, B esso (Milano), I aly and Ins i u e o Biomedical Technologies Na ional
Cen e o Resea ch Seg a e, Milano, I aly. 204 Depa men o Public Heal h & Clinical Medicine, Umeå Uni e si y, Umeå, Sweden. 205 Depa men o
Genomics o Common Disease, Impe ial College London, London, UK. 206 Ca olina Popula ion Cen e , Uni e si y o No h Ca olina a Chapel Hill, Chapel Hill,
No h Ca olina 27516, USA. 207 Depa men o Psychia y and Psycho he apy, Uni e si y Medicine G ei swald, G ei swald, Ge many. 208 Ge man Cen e o
Neu odegene a i e Diseases (DZNE), Ros ock and G ei swald Si e, G ei swald, Ge many. 209 Depa men o P e en i e Medicine, No is Comp ehensi e
Cance Cen e , Keck School o Medicine, Uni e si y o Sou he n Cali o nia, Los Angeles, Cali o nia 90089, USA. 210 Du e Cen e o Ca diogene ic Resea ch,
Ams e dam, The Ne he lands. 211 In e uni e si y Ca diology Ins i u e o he Ne he lands, U ech , The Ne he lands. 212 Facul y o Med, Na ional Hea & Lung
Ins i u e, Ca dio ascula Science, Hamme smi h Campus, Hamme smi h Hospi al, Hamme smi h Campus, Impe ial College London, London, UK. 213 Synlab
Academy, Synlab Se ices GmbH, Mannheim, Ge many. 214 Clinical Ins i u e o Medical and Chemical Labo a o y Diagnos ics, Medical Uni e si y o G az,
G az, Aus ia. 215 Ox o d Na ional Ins i u e o Heal h Resea ch (NIHR) Biomedical Resea ch Cen e, Chu chill Hospi al, Ox o d, UK. 216 Depa men o
Bios a is ics, Uni e si y o Li e pool, Li e pool L69 3GL, UK. 217 School o Public Heal h, Uni e si y o Adelaide, Adelaide, Sou h Aus alia 5005, Aus alia.
218 Labo a o y o Clinical Chemis y and Hema ology, Di ision Labo a o ies & Pha macy, UMC U ech , U ech , The Ne he lands. 219 Depa men o
Medicine, Uni e si y o Washing on, Sea le, Washing on 98195, USA. 220 Depa men o Epidemiology, Uni e si y o Washing on, Sea le, Washing on
98101, USA. 221 G oup Heal h Resea ch Ins i u e, G oup Heal h Coope a i e, Sea le, Washing on 98101, USA. 222 Depa men o Epidemiology, School o
Public Heal h and T opical Medicine, Tulane Uni e si y, New O leans, Louisiana, USA. 223 Depa men o Clinical Physiology and Nuclea Medicine, Kuopio
Uni e si y Hospi al, Kuopio, Finland. 224 Di ision o Popula ion Heal h Sciences, Ninewells Hospi al and Medical School, Uni e si y o Dundee, Dundee, DD2
4RB, Sco land. 225 Depa men o Clinical Epidemiology ( o me ly Ins i u e o P e en i e Medicine), Bispebje g and F ede iksbe g Hospi al (2000
F ede iksbe g), The Capi al Region, Copenhagen, Denma k. 226 MRC In eg a i e Epidemiology Uni , B is ol Uni e si y, B is ol, UK. 227 Depa men o
Psychia y E asmus Medical Cen e , Ro e dam, The Ne he lands. 228 Du e Cen e o Ca diogene ic Resea ch, ICIN-Ne he lands Hea Ins i u e, U ech ,
The Ne he lands. 229 Ushe Ins i u e o Popula ion Heal h Sciences and In o ma ics, The Uni e si y o Edinbu gh, Sco land, UK. 230 Gene ics o Complex
T ai s, Uni e si y o Exe e Medical School, RILD Building Uni e si y o Exe e , Exe e EX2 5DW, UK. 231 Eu opean Cen e o En i onmen and Human Heal h,
Uni e si y o Exe e Medical School, The Knowledge Spa, T u o TR1 3HD, UK. 232 Gene ics o Complex T ai s, Uni e si y o Exe e Medical School, Uni e si y
o Exe e , Exe e EX1 2LU, UK. 233 Wellcome T us Sange Ins i u e, Hinx on, Camb idge, UK. 234 NIHR Camb idge Biomedical Resea ch Cen e, Le el 4,
Ins i u e o Me abolic Science Box 289 Addenb ooke’s Hospi al, Camb idge CB2 OQQ, UK. 235 Uni e si y o Camb idge Me abolic Resea ch Labo a o ies,
Le el 4, Ins i u e o Me abolic Science Box 289 Addenb ooke’s Hospi al, Camb idge CB2 OQQ, UK. 236 P incess Al-Jawha a Al-B ahim Cen e o Excellence
in Resea ch o He edi a y Diso de s (PACER-HD), King Abdulaziz Uni e si y, Jeddah, Saudi A abia. 237 Depa men o Gene ics, Ha a d Medical School,
Bos on Massachuse s 02115, USA. 238 Depa men o Epidemiology, Ha a d T.H. Chan School o Public Heal h, Bos on, Massachuse s 02115, USA.
239 Channing Di ision o Ne wo k Medicine, Depa men o Medicine, B igham and Women’s Hospi al and Ha a d Medical School, Bos on, Massachuse s
02115, USA. 240 Di ision o Popula ion Heal h Sciences and Educa ion, S Geo ge’s, Uni e si y o London, London SW17 0RE, UK. 241 Ne he lands Genomics
ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14977
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Ini ia i e (NGI)-sponso ed Ne he lands Conso ium o Heal hy Aging (NCHA). Leiden, The Ne he lands. 242 Cen e o Medical Sys ems Biology, Leiden, The
Ne he lands. 243 Ins i u e o Gene ic Epidemiology, Helmhol z Zen um Mu
¨nchen - Ge man Resea ch Cen e o En i onmen al Heal h, Neuhe be g 85764,
Ge many. 244 Depa men o S a is ics, Uni e si y o Ox o d, Ox o d, UK. 245 Moun Sinai School o Medicine, New Yo k 10029, USA. 246 The Mindich Child
Heal h and De elopmen Ins i u e, Icahn School o Medicine a Moun Sinai, New Yo k, New Yo k 10029, USA. 247 Depa men o P e en i e Medicine, The
Icahn School o Medicine a Moun Sinai, New Yo k, New Yo k 10029, USA. * These au ho s con ibu ed equally o his wo k. ** These au ho s join ly
supe ised his wo k.
zDeceased.
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