Discovery of novel heart rate-associated loci using the Exome Chip
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ASSOCIATION STUDIES ARTICLE
Disco e y o no el hea a e-associa ed loci using he
Exome Chip
Ma en E. an den Be g
1,†
, Helen R. Wa en
2,3,†
, Claudia P. Cab e a
2,3
, Niek
Ve weij
4
, Bo bala Mi sud
2,3
, Je ey Haessle
5
, Na han A. Bihlmeye
6
, Yi-Ping
Fu
7
, S e an Weiss
8,9
, Hen y J. Lin
10,11
, Niels G a up
12
, Rui ang Li-Gao
13
,
Gio gio Pis is
14,15
, Nabi Shah
16,17
, Jenni e A. B ody
18
,
Ma ina Mu¨ lle -Nu asyid
19,20,21
, Honghuang Lin
22
, Hao Mei
23
, Albe V.
Smi h
24,25
, Leo-Pekka Lyy ik€
ainen
26
, Leanne M. Hall
27,28
, Jessica an Se en
29
,
S ella T ompe
30,31
, B am P. P ins
32,33
, Aa on Isaacs
34
, Fa id Radmanesh
35,36
,
Jona han Ma en
37
, Aiman En wis le
2,3
, Jan A. Ko s
1
, Claudia T. Sil a
38,39,40
,
Al a o Alonso
41
, Joshua C. Bis
18
, Rudol de Boe
4
, Hugoline G. de Haan
13
,
Rene´e de Mu se
13
, Geo ge Dedoussis
42
, Anna F. Dominiczak
43
, Alex S. F.
Doney
16
, Pa ick T. Ellino
44,36
, Ruben N. Eppinga
4
, S ephan B. Felix
45
, Xiuqing
Guo
10
, Yanick Hagemeije
4
, To ben Hansen
12
, Tama a B. Ha is
46
, Susan R.
Heckbe
47,48
, Paul L. Huang
44
, Shih-Jen Hwang
49
, Mika K€
aho¨nen
50
, Jø gen K.
Kan e s
51
, I ana Kolcic
52
, Leno e J. Laune
46
, Man Li
53
, Jie Yao
10
, Allan
Linnebe g
54,55,56
, Simin Liu
57
, Pe e W. Mac a lane
58
, Massimo Mangino
59,60
,
And ew D. Mo is
61
, An onella Mulas
14
, Alison D. Mu ay
62
, Ch is ophe P.
Nelson
27,28
, Ma co O
u
63
, Sandosh Padmanabhan
43,64
, Anne e Pe e s
65,20,66
,
Da id J. Po eous
67
, Neil Poul e
68
, B uce M. Psa y
69,70
, Lihong Qi
71
, Olli T.
Rai aka i
72
, Fe nando Ri adenei a
73
, Ca olina Roselli
74
, Igo Rudan
61
, Na eed
Sa a
64
, Pe e Se e
75
, Mo i z F. Sinne
20,21
, Elsayed Z. Soliman
76
, Timo hy D.
Spec o
59
, Alice V. S an on
77
, Ka hleen E. S i ups
2,78
, Ken D. Taylo
79,80,81
,
Ma in D. Tobin
82
, And e´ Ui e linden
83
, Ilonca Vaa jes
84
, A no W. Hoes
85
,
Pe e an de Mee
4
, Uwe Vo¨lke
8,9
, Melanie Waldenbe ge
61,65,20
,
†
The au ho s wish i o be known ha in hei opinion, he i s wo au ho s should be ega ded as join i s au ho s because o equal con ibu ion.
Recei ed: Janua y 16, 2017. Re ised: Ma ch 13, 2017. Accep ed: Ma ch 18, 2017
V
CThe Au ho 2017. Published by Ox o d Uni e si y P ess.
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 (h p://c ea i ecommons.o g/licenses/by/4.0/),
which pe mi s un es ic ed euse, dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed.
2346
Human Molecula Gene ics, 2017, Vol. 26, No. 12 2346–2363
doi: 10.1093/hmg/ddx113
Ad ance Access Publica ion Da e: 3 Ap il 2017
Associa ion S udies A icle
Zhijun Xie
22
, Magdalena Zoledziewska
14
, And ew Tinke
2,3
, Oz en
Polasek
52,61
, Jona han Rosand
35,36
, Yalda Jamshidi
33
, Co nelia M. an Duijn
38
,
Ele he ia Zeggini
32
, J. Wou e Jukema
30
, Folke W. Asselbe gs
29,86,87
, Nilesh
J. Samani
27,28
, Te ho Leh im€
aki
88
, Vilmundu Gudnason
24,25
, James Wilson
89
,
S e en A. Lubi z
44,36
, S e an K€
a€
ab
21,20
, Nona So oodehnia
90
,
Ma k J. Caul ield
2,3
, Colin N. A. Palme
16
, Se ena Sanna
14
,
Dennis O. Mook-Kanamo i
13,91
, Panos Deloukas
2
, Olu Pede sen
12
, Je ome I.
Ro e
92
, Ma cus Do¨
45
, Ch is J. O’Donnell
93
, Ca oline Haywa d
37
, Dan E.
A king
94
, Cha les Koope be g
5
, Pim an de Ha s
4
, Ma k Eijgelsheim
95
,
B uno H. S icke
95
and Pa icia B. Mun oe
2,3,
*
1
Depa men o Medical In o ma ics E asmus MC - Uni e si y Medical Cen e Ro e dam, P.O. Box 2040, 3000CA,
Ro e dam, he Ne he lands,
2
Clinical Pha macology, William Ha ey Resea ch Ins i u e, Queen Ma y
Uni e si y o London, London, EC1M 6BQ, UK,
3
NIHR Ba s Ca dio ascula Biomedical Resea ch Uni , Queen
Ma y Uni e si y o London, London, EC1M 6BQ, UK,
4
Uni e si y Medical Cen e G oningen, Uni e si y o
G oningen, Depa men o Ca diology, he Ne he lands,
5
Di ision o Public Heal h Sciences, F ed Hu chinson
Cance Resea ch Cen e , Sea le, WA 98109, USA,
6
P edoc o al T aining P og am in Human Gene ics, McKusick-
Na hans Ins i u e o Gene ic Medicine, Johns Hopkins Uni e si y School o Medicine, Bal imo e, MD, USA,
21205,
7
Di ision o Ca dio ascula Sciences, Na ional Hea , Lung, and Blood Ins i u e, Na ional Ins i u es o
Heal h, Be hesda, MD, USA ,
8
In e acul y Ins i u e o Gene ics and Func ional Genomics; Uni e si y Medicine
and E ns -Mo i z-A nd -Uni e si y G ei swald; G ei swald, 17475, Ge many,
9
DZHK (Ge man Cen e o
Ca dio ascula Resea ch); pa ne si e G ei swald; G ei swald, 17475, Ge many,
10
The Ins i u e o
T ansla ional Genomics and Popula ion Sciences, Depa men o Pedia ics, Los Angeles Biomedical Resea ch
Ins i u e a Ha bo -UCLA Medical Cen e , 1124 W. Ca son S ee , To ance, CA 90502, USA,
11
Di ision o Medical
Gene ics, Depa men o Pedia ics, Ha bo -UCLA Medical Cen e , To ance, CA, USA,
12
The No o No disk
Founda ion Cen e o Basic Me abolic Resea ch, Facul y o Heal h and Medical Sciences, Uni e si y o
Copenhagen, Copenhagen, Denma k,
13
Depa men o Clinical Epidemiology, Leiden Uni e si y Medical Cen e ,
Leiden, he Ne he lands,
14
Is i u o di Rice ca Gene ica e Biomedica (IRGB), CNR, Monse a o, I aly ,
15
Cen e o
S a is ical Gene ics, Uni e si y o Michigan, Ann A bo , MI, USA,
16
Di ision o Molecula and Clinical Medicine,
School o Medicine, Uni e si y o Dundee, DD1 9SY, UK,
17
Depa men o Pha macy, COMSATS Ins i u e o
In o ma ion Technology, Abbo abad, 22060, Pakis an,
18
Ca dio ascula Heal h Resea ch Uni , Depa men o
Medicine, Uni e si y o Washing on, 1730 Mino A e, Sui e 1360, Sea le, WA 98101, USA,
19
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,
Ge many,
20
DZHK (Ge man Cen e o Ca dio ascula Resea ch), pa ne si e Munich Hea Alliance, Munich,
Ge many,
21
Depa men o Medicine I, Uni e si y Hospi al Munich, Ludwig-Maximilians-Uni e si €
a , Munich,
Ge many,
22
Sec ion o Compu a ional Biomedicine, Depa men o Medicine, Bos on Uni e si y School o
Medicine, Bos on, MA,
23
Depa men o Da a Science, Uni e si y o Mississippi Medical Cen e , Jackson, MI,
USA,
24
Icelandic Hea Associa ion, 201 Kopa ogu , Iceland,
25
Facul y o Medicine, Uni e si y o Iceland, 101
Reykja ik, Iceland,
26
Depa men o Clinical Chemis y, Fimlab Labo a o ies and Uni e si y o Tampe e School
o Medicine, A o, D339, P.O. Box 100, FI-33014 Tampe e, Finland,
27
Depa men o Ca dio ascula Sciences,
Uni e si y o Leices e , Ca dio ascula Resea ch Cen e, Glen ield Hospi al, Leices e , LE3 9QP, UK,
28
NIHR
Leices e Ca dio ascula Biomedical Resea ch Uni , Glen ield Hospi al, Leices e LE3 9QP, UK,
29
Depa men o
Ca diology, Di ision Hea & Lungs, Uni e si y Medical Cen e U ech , U ech , he Ne he lands,
30
Depa men
o Ca diology, Leiden Uni e si y Medical Cen e , 2300 RC, Leiden, he Ne he lands,
31
Depa men o
Ge on ology and Ge ia ics, Leiden uni e si y Medical Cen e , Leiden, he Ne he lands,
32
Depa men o
Human Gene ics, Wellcome T us Sange Ins i u e, Hinx on, Uni ed Kingdom, CB10 1SA,
33
Ca diogene ics Lab,
Gene ics and Molecula Cell Sciences Resea ch Cen e, Ca dio ascula and Cell Sciences Ins i u e, S Geo ge’s,
2347Human Molecula Gene ics, 2017, Vol. 26, No. 12 |
Uni e si y o London, C anme Te ace, London, SW17 0RE, UK,
34
CARIM School o Ca dio ascula Diseases,
Maas ich Cen e o Sys ems Biology (MaCSBio), Dep . o Biochemis y, Maas ich Uni e si y,
Uni e si ei ssingel 60, 6229 ER Maas ich , NL,
35
Cen e o Human Gene ic Resea ch, Massachuse s Gene al
Hospi al, Bos on, MA 02114,
36
P og am in Medical and Popula ion Gene ics, B oad Ins i u e, Camb idge, MA
02142,
37
MRC Human Gene ics Uni , MRC Ins i u e o Gene ics and Molecula Medicine, Uni e si y o
Edinbu gh, Wes e n Gene al Hospi al, C ewe Road Sou h, Edinbu gh, EH4 2XU, UK,
38
Gene ic Epidemiology
Uni , Dep . o Epidemiology, E asmus Uni e si y Medical Cen e , PO Box 2040, 3000 CA Ro e dam, NL,
39
Doc o al P og am in Biomedical Sciences, Uni e sidad del Rosa io, Bogo
a, Colombia,
40
GENIUROS G oup,
Gene ics and Genomics Resea ch Cen e CIGGUR, School o Medicine and Heal h Sciences, Uni e sidad del
Rosa io, Bogo
a, Colombia,
41
Depa men o Epidemiology, Rollins School o Public Heal h, Emo y Uni e si y,
A lan a, GA, 30322,
42
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 17671, G eece,
43
Ins i u e o Ca dio ascula and Medical Sciences, College o Medical,
Ve e ina y and Li e Sciences, Uni e si y o Glasgow, Glasgow, UK,
44
Ca dio ascula Resea ch Cen e ,
Massachuse s Gene al Hospi al, Cha les own, MA, USA,
45
Depa men o In e nal Medicine B - Ca diology,
Pneumology, In ec ious Diseases, In ensi e Ca e Medicine; Uni e si y Medicine G ei swald; G ei swald, 17475,
Ge many & DZHK (Ge man Cen e o Ca dio ascula Resea ch); pa ne si e G ei swald; G ei swald, 17475,
Ge many,
46
Labo a o y o Epidemiology and Popula ion Sciences, Na ional Ins i u e on Aging, In amu al
Resea ch P og am, Na ional Ins i u es o Heal h, Be hesda, Ma yland, 20892, USA,
47
Ca dio ascula Heal h
Resea ch Uni and Depa men o Epidemiology, Uni e si y o Washing on, 1730 Mino A e, Sui e 1360, Sea le,
WA 98101, USA,
48
G oup Heal h Resea ch Ins i u e, G oup Heal h Coope a i e, 1730 Mino A e, Sui e 1600,
Sea le, WA, USA,
49
Popula ion Sciences B anch, Di ision o In amu al Resea ch, NHLBI, NIH, Be hesda MD,
USA,
50
Depa men o Clinical Physiology, Tampe e Uni e si y Hospi al and Uni e si y o Tampe e School o
Medicine, Finn-Medi 1, 3 h loo , P.O. Box 2000, FI-33521 Tampe e, Finland,
51
Labo a o y o Expe imen al
Ca diology, Uni e si y o Copenhagen, Copenhagen, Denma k,
52
Facul y o Medicine, Uni e si y o Spli , Spli ,
C oa ia,
53
Di ision o Neph ology & Hype ension, In e nal Medicine, School o Medicine, Uni e si y o U ah,
Sal Lake Ci y, UT 84109, USA,
54
Resea ch Cen e o P e en ion and Heal h, Capi al Region o Denma k,
Copenhagen, Denma k,
55
Depa men o Clinical Expe imen al Resea ch, Rigshospi ale , Glos up, Denma k,
56
Depa men o Clinical Medicine, Facul y o Heal h and Medical Sciences, Uni e si y o Copenhagen,
Copenhagen, Denma k,
57
B own Uni e si y School o Public Heal h, P o idence, Rhode Island 02912, USA,
58
Ins i u e o Heal h and Wellbeing, Uni e si y o Glasgow, Glasgow, UK,
59
Depa men o Twin Resea ch and
Gene ic Epidemiology, King’s College London, London, UK,
60
NIHR Biomedical Resea ch Cen e a Guy’s and S
Thomas’ Founda ion T us , London SE1 9RT, UK,
61
Ushe Ins i u e o Popula ion Heal h Sciences and
In o ma ics, Uni e si y o Edinbu gh, Edinbu gh, EH8 9AG, UK,
62
Abe deen Biomedical Imaging Cen e, Lilian
Su on Building, Uni e si y o Abe deen, Fo es e hill, Abe deen AB25 2ZD, UK,
63
Uni a Ope a i a Complessa di
Ca diologia, P esidio Ospedalie o Oncologico A mando Businco Caglia i , Azienda Ospedalie a B o zu Caglia i,
Cagllia i, I aly,
64
Ins i u e o Ca dio ascula and Medical Sciences, Uni e si y o Glasgow, BHF GCRC, Glasgow
G12 8TA, UK,
65
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, Neuhe be g, Ge many,
66
Ge man Cen e o Diabe es Resea ch, Neuhe be g, Ge many,
67
Cen e o Genomic & Expe imen al Medicine, Ins i u e o Gene ics & Molecula Medicine, Uni e si y o
Edinbu gh, Wes e n Gene al Hospi al, C ewe Road Sou h, Edinbu gh EH4 2XU, UK,
68
School o Public Heal h,
Impe ial College London, W2 1PG, UK,
69
Ca dio ascula Heal h Resea ch Uni , Depa men o Heal h Se ices,
Uni e si y o Washing on, 1730 Mino A e, Sui e 1360, Sea le, WA 98101, USA,
70
G oup Heal h Resea ch
Ins i u e, G oup Heal h Coope a i e, Sea le, WA, USA,
71
Uni e si y o Cali o nia Da is, One Shields A e Ms1c
145, Da is, CA 95616 USA,
72
Depa men o Clinical Physiology and Nuclea Medicine, Tu ku Uni e si y
Hospi al, and Resea ch Cen e o Applied and P e en i e Ca dio ascula Medicine, Uni e si y o Tu ku, P.O.
Box 52, FI-20521 Tu ku, Finland,
73
Human Genomics Facili y E asmus MC - Uni e si y Medical Cen e
Ro e dam, P.O. Box 2040, 3000CA, Ro e dam, he Ne he lands,
74
P og am in Medical and Popula ion Gene ics,
B oad Ins i u e o MIT and Ha a d, Camb idge, MA, USA,
75
Na ional Hea and Lung Ins i u e, Impe ial College
London, W2 1PG, UK,
76
Epidemiological Ca diology Resea ch Cen e (EPICARE), Wake Fo es School o Medicine,
Wins on-Salem, NC 27157, USA,
77
Molecula and Cellula The apeu ics, Royal College o Su geons in I eland,
Dublin 2, I eland,
78
Depa men o Haema ology, Uni e si y o Camb idge, Camb idge, UK,
79
Ins i u e o
2348 |Human Molecula Gene ics, 2017, Vol. 26, No. 12
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, CA, USA,
80
Di ision o Genomic Ou comes, Depa men o Pedia ics, Ha bo -UCLA
Medical Cen e , To ance, CA, USA,
81
Depa men s o Pedia ics, Medicine, and Human Gene ics, UCLA, Los
Angeles, CA, USA,
82
Depa men o Heal h Sciences, Uni e si y o Leices e , Leices e LE1 7RH, UK,
83
Human
Geno yping Facili y E asmus MC - Uni e si y Medical Cen e Ro e dam, P.O. Box 2040, 3000CA, Ro e dam, he
Ne he lands,
84
Julius Cen e o Heal h Sciences and P ima y Ca e, Uni e si y Medical Cen e , PO Box 85500,
3508 GA U ech , he Ne he lands,
85
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, Neuhe be g, Ge many,
86
Du e Cen e o Ca diogene ic
Resea ch, ICIN-Ne he lands Hea Ins i u e, U ech , he Ne he lands,
87
Ins i u e o Ca dio ascula Science,
Facul y o Popula ion Heal h Sciences, Uni e si y College London, London, UK,
88
Depa men o Clinical
Chemis y, Fimlab Labo a o ies and Uni e si y o Tampe e School o Medicine, A o, D338, P.O. Box 100, FI-
33014 Tampe e, Finland,
89
Physiology & Biophysics, Uni e si y o Mississippi Medical Cen e , Jackson, MI, USA,
90
Ca dio ascula Heal h Resea ch Uni , Di ision o Ca diology, Depa men s o Medicine and Epidemiology,
Uni e si y o Washing on, 1730 Mino A e, Sui e 1360, Sea le, WA 98101, USA,
91
Depa men o Public Heal h
and P ima y Ca e, Leiden Uni e si y Medical Cen e , Leiden, he Ne he lands,
92
The Ins i u e o T ansla ional
Genomics and Popula ion Sciences, Depa men s o Pedia ics and Medicine, Los Angeles Biomedical Resea ch
Ins i u e a Ha bo -UCLA Medical Cen e , 1124 W. Ca son S ee , To ance, CA 90502, USA,
93
Bos on Ve e an’s
Adminis a ion Heal hca e, Bos on MA, USA,
94
McKusick-Na hans Ins i u e o Gene ic Medicine, Johns Hopkins
Uni e si y School o Medicine, Bal imo e, MD, USA, 21205 and
95
Depa men o Epidemiology E asmus MC -
Uni e si y Medical Cen e Ro e dam, P.O. Box 2040, 3000CA, Ro e dam, he Ne he lands
*To whom co espondence should be add essed a : Depa men o Clinical Pha macology, 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, Cha e house Squa e, London EC1M 6BQ, UK. Tel: þ44 2078823586; Fax:
þ442078823408; Email: [email p o ec ed]
Abs ac
Res ing hea a e is a he i able ai , and an inc ease in hea a e is associa ed wi h inc eased mo ali y isk. Genome-wide
associa ion s udy analyses ha e ound loci associa ed wi h es ing hea a e, a he ime o ou s udy hese loci explained
0.9% o he a ia ion. This s udy aims o disco e new gene ic loci associa ed wi h hea a e om Exome Chip me a-
analyses.
Hea a e was measu ed om ei he elec oca diog ams o pulse eco dings. We me a-analysed hea a e associa ion e-
sul s om 104 452 Eu opean-ances y indi iduals om 30 coho s, geno yped using he Exome Chip. Twen y- ou a ian s
we e selec ed o ollow-up in an independen da ase (UK Biobank, N¼134 251). Condi ional and gene-based es ing was
unde aken, and a ian s we e in es iga ed wi h bioin o ma ics me hods.
We disco e ed i e no el hea a e loci, and one new independen low- equency non-synonymous a ian in an es ablished
hea a e locus (KIAA1755). Lead a ian s in ou o he no el loci a e non-synonymous a ian s in he genes C10o 71,
DALDR3, TESK2 and SEC31B. The a ian a SEC31B is signi ican ly associa ed wi h SEC31B exp ession in hea and ibial ne e
issue. Fu he candida e genes we e de ec ed om long- ange egula o y ch oma in in e ac ions in hea issue (SCD,SLF2
and MAPK8). We obse ed signi ican en ichmen in DNase I hype sensi i e si es in e al hea and lung. Mo eo e , en ich-
men was seen o he i s ime in human neu onal p ogeni o cells (de i ed om emb yonic s em cells) and e al muscle
samples by including ou no el a ian s.
Ou indings ad ance he knowledge o he gene ic a chi ec u e o hea a e, and indica e new candida e genes o ollow-up
unc ional s udies.
In oduc ion
Inc eased es ing hea a e (HR) is a known isk ac o o ca -
dio ascula mo bidi y and mo ali y (1–3), including s oke (4)
and sudden ca diac dea h (5,6). Hea a e inc eased by 20 bea s
pe minu e (BPM) is associa ed wi h 30-50% highe mo ali y
and appea s o be independen o con ounde ac o s (7). High
HR inc eases myoca dial oxygen consump ion ye lessens oxy-
gen deli e y o myoca dial issue. I also inc eases a e ial s i -
ness and isk o plaque up u e (8). Al hough HR can be
in luenced by many non-gene ic ac o s (e.g. exe cise, smoking
and ca dio ascula d ugs), he he i abili y o es ing HR is es i-
ma ed o be 26–32% om amily s udies (9,10), and 55–63% om
win s udies (11).
Se e al me a-analyses o genome-wide associa ion s udies
(GWASs) ha e been unde aken o de ec gene ic de e minan s
o HR (12–16). The e we e 21 HR loci p e iously epo ed a he
ime o ou s udy by den Hoed e al. (12) in a GWAS analysis o
180 000 indi iduals, p edominan ly o Eu opean ances y. The
s udy implica ed 20 candida e genes om ollow-up unc ional
2349Human Molecula Gene ics, 2017, Vol. 26, No. 12 |
s udies in Danio e io and D osophila melanogas e models.
Smalle GWAS analyses ha e also been pe o med in Icelandic
and No wegian popula ions (15), A ican Ame icans (13) and
gene ically isola ed Eu opean popula ions (16). The a ian s dis-
co e ed by GWAS a e common, and a e mos ly in in ons o
in e genic egions. Toge he he p e ious loci om GWAS a he
ime o ou s udy only explain a small pe cen age [0.9% o he
a iabili y in HR (12,17)].
To inc ease ou knowledge o gene ic de e minan s in lu-
encing HR and disco e no el loci, especially a e o low e-
quency coding a ian s wi h la ge e ec s, we me a-analysed
da a om 104 452 indi iduals o Eu opean-ances y using he
Exome Chip, om coho s ha pa icipa ed in he Coho s o
Hea & Aging Resea ch in Genomic Epidemiology (CHARGE)
EKG conso ium. The Exome Chip pe mi s a cos -e icien
analysis o coding a ian s de i ed om sequencing
o >12 000 indi iduals and includes many a e and low- e-
quency a ian s (18). We pe o med a alida ion expe imen
using independen eplica ion samples om UK Biobank
da a, and bioin o ma ics in es iga ions o gain an unde -
s anding o he new HR loci.
Resul s
Single-nucleo ide a ian analysis in indi iduals o
Eu opean-ances y
In he disco e y phase, associa ion esul s o 235 677 single-nu-
cleo ide a ian s (SNVs) om 104 452 indi iduals we e me a-
analysed using a ixed-e ec s model (Supplemen a y Ma e ial,
Fig. S1). Two analyses we e pe o med. The i s used RR-in e -
als (RR in milliseconds¼60 000/HR, in bea s pe minu e, ac-
co ding o he in e se ela ionship be ween HR and RR). The
second used he in e se-no malized esiduals o he linea e-
g ession RR-in e al adjus ed o age þsex þbody mass index
(BMI) as co a ia es (deno ed as RR-INVN). An o e iew o he
s udy design is p o ided in Figu e 1.
We obse ed a high co ela ion o e ec sizes and P- alues
be ween he RR-in e al and RR-INVN me a-analyses (
2
¼0.99
and 0.98, espec i ely; Supplemen a y Ma e ial, Fig. S2).
Fu he mo e, he RR-in e al was nea -no mally dis ibu ed,
so in e se no maliza ion was deemed unnecessa y
(Supplemen a y Ma e ial, Fig. S3).
Be a-blocke s a e clinically known o lowe HR, he e o e
he pheno ype measu emen s o be a-blocke use s may be
unde -es ima ed, and hence he inclusion o be a-blocke
use s in ou analysis may po en ially bias ou analysis esul s.
We he e o e pe o med a sensi i i y analysis by also me a-
analysing a subg oup o coho s ha p o ided be a-blocke
da a (N¼48 347; 17 coho s). Resul s including o excluding
be a-blocke use s we e highly co ela ed (
2
o he
be as ¼0.97;
2
o he P- alues ¼0.74; Supplemen a y Ma e ial,
Fig. S4), sugges ing he e is li le o no bias om including
be a-blocke use s in he analysis. The e o e we epo he
me a-analysis esul s om he ull da ase o he RR-in e al,
o maximize sample size and powe .
Replica ion and me a-analysis wi h he UK Biobank
da ase
To iden i y no el associa ed loci, we selec ed 12 a ian s wi h
P<110
5
ha mapped ou side he 21 HR loci epo ed in he
p e ious GWAS (12) o ollow-up in an independen da ase .
Wi hin each unknown locus, he e we e no po en ial seconda y
SNVs no in linkage disequilib ium (LD) wi h he lead SNV
(
2
<0.2) and mee ing ou look-up signi icance h eshold
(P<110
5
). Hence only 12 new lead SNVs we e ca ied o -
wa d. We also ollowed-up 12 po en ial seconda y signals a 9 o
he 21 p e iously epo ed HR loci ( u he de ails on selec ion
c i e ia a e p o ided in he Ma e ials and Me hods) (12). None o
he selec ed a ian s was in LD (
2
<0.2) wi h each o he , o wi h
he published SNVs. Thus, a o al o 24 a ian s we e aken o -
wa d in o eplica ion. The UK Biobank da ase p o ided esul s
o he selec ed gene ic a ian s (N¼134 251 indi iduals).
Nine o he 12 p e iously unknown a ian s we e alida ed
based on exome-wide signi icance (P2.12 10
7
) in he com-
bined me a-analysis o CHARGE and UK Biobank da a, and on
Bon e oni-adjus ed signi icance (P0.0042 o 12 es s) in he
eplica ion da ase alone, wi h conco dan di ec ions o e ec s
aking in o accoun he in e se ela ionship be ween he RR-
in e al om he disco e y da a and HR om he eplica ion
da a (Table 1;Fig. 2). Indeed, all nine SNV associa ions we e
genome-wide signi ican in he combined me a-analysis
(P<5.0 10
8
). Fou o ou nine alida ed no el loci we e e-
po ed in a UK Biobank s udy (17) ha was published a e com-
ple ion o ou s udy (Table 1B). Hence, we p esen esul s he e
o i e un epo ed no el loci (Table 1A; Supplemen a y
Ma e ial, Figs S5 and S6).
Twel e o he 21 HR-associa ed SNVs om he p e iously e-
po ed GWAS (12) we e co e ed on he Exome Chip, ei he di -
ec ly o by a p oxy SNV in high LD (
2
>0.8). Ou disco e y me a-
analysis showed s ong suppo o he p e ious indings, wi h
11 o he 12 SNVs alida ed a Bon e oni-adjus ed signi icance
(P0.0042 o 12 es s), o which nine we e alida ed a exome-
wide signi icance (P<210
7
;Fig. 2). Only s4140885 a he TFPI
locus was no suppo ed in ou da a (P¼0.10; Supplemen a y
Ma e ial, Table S1).
Independen seconda y signals a known loci
All 12 po en ial seconda y signals a loci p e iously epo ed by
den Hoed e al. (12) we e genome-wide signi ican in he com-
bined me a-analysis (Supplemen a y Ma e ial, Table S2) and a e
independen o he known SNPs acco ding o LD (
2
<0.2). We
pe o med a condi ional analysis using Genome-wide Complex
T ai s Analysis (GCTA) o o mally iden i y seconda y signals o
associa ion. Fi e o he 12 alida ed po en ial seconda y SNVs
(wi hin CD46,CCDC141,SLC35F1,ACHE and KIAA1755 loci) we e
selec ed wi hin he inal GCTA model (Supplemen a y Ma e ial,
Table S3). A ou o he p e iously epo ed HR egions he sec-
onda y signals ha we iden i ied we e con i med o be s a is ic-
ally independen signals o associa ion: CD46 ( s2745967),
CCDC141 ( s10497529), SLC35F1 ( s12210810) and KIAA1755
( s41282820) in addi ion o he known SNV, as bo h he pub-
lished SNV and he new seconda y SNV we e p esen in he
inal GCTA model o join ly independen associa ed a ian s.
Hence, we iden i ied wo dis inc signals o associa ion a each
o hese ou known HR loci. Howe e , he published SNV a he
ACHE locus ( s13245899) is no co e ed on he Exome Chip, o by
any p oxies (Supplemen a y Ma e ial, Table S1), so he GCTA
analysis does no include he known a ian . As we a e no able
o condi ion on he una ailable published SNV and o mally
es associa ion join ly wi h he known SNV, we a e unable o
s a is ically con i m he o al numbe o independen signals a
he ACHE locus.
The seconda y SNVs a CCDC141, ACHE and KIAA1755 a e
non-synonymous a ian s. Fu he mo e, he SNVs a CCDC141
2350 |Human Molecula Gene ics, 2017, Vol. 26, No. 12
and KIAA1755 a e low- equency wi h mino allele equencies
(MAFs) o 3.6 and 1.7%, espec i ely. Seconda y signals ha e
also ecen ly been obse ed a ou o he i e loci (CD46,
CCDC141,SLC35F1 and ACHE) in UK Biobank da a (17), since
comple ion o ou me a-analysis. A CD46, ou seconda y SNV
( s2745967) is in high LD (
2
¼0.78) wi h he seconda y SNV
( s2745959) epo ed in UK Biobank, so likely o be he same sig-
nal. A CCDC141 ou seconda y a ian is exac ly he same SNV
as om UK Biobank ( s10497529). Simila ly, a SLC35F1, ou sec-
onda y SNV ( s12210810) is in e y high LD (
2
¼0.98), so is likely
o be he same signal. Hence a hese h ee known loci (CD46,
CCDC141,SLC35F1), all exis ing da a sugges he e a e wo inde-
penden signals o associa ion. A he ACHE locus, ou second-
a y SNV ( s542137; 38 kb and
2
<0.2 om he published SNV)
is no in LD (
2
<0.2) wi h he seconda y SNV om UK Biobank
( s140367586; 659 kb and
2
<0.2 om he published SNV). We
a e unable o clea ly de e mine he numbe o dis inc signals a
he ACHE locus om ou Exome Chip RR-in e al disco e y
me a-analysis da a, wi hou he published SNV being co e ed
on he Exome Chip. The low- equency non-synonymous a i-
an ( s41282820) a he known KIAA1755 locus is a new, second-
a y a ian , wi h s ong e idence o independen associa ion, i
does no o e lap wi h o he published indings.
Va iance explained
Twel e o he 21 p e iously epo ed HR-associa ed SNVs (12)
co e ed on he Exome Chip explain 1.14% o RR-in e al a i-
ance (P¼3.96 10
10
) wi hin he 1958 Bi h Coho s udy (see
Ma e ials and Me hods). The added con ibu ion o he lead
SNVs a ou i e un epo ed no el loci, combined wi h he 12
Figu e 1. Schema ic low diag am o he s udy design. N, sample size; SKAT, SNV-se Ke nel Associa ion Tes ; P,P- alue; LD, linkage disequilib ium; SNV, single nucleo-
ide a ian ; GCTA, Genome-wide Complex T ai s Analysis so wa e; 1958BC, 1958 Bi h Coho ; UKB, UK Biobank.
2351Human Molecula Gene ics, 2017, Vol. 26, No. 12 |
p e iously epo ed SNVs, inc eases he a iance explained o
1.28% o e all (P¼9.17 10
11
).
Compa ison o esul s be ween Eu opean and
non-Eu opean popula ions
To in es iga e ou da a om non-Eu opean samples [9358
A ican Ame icans (AA), 1411 Hispanic (HIS) and 754
Chinese-Ame icans (CH); Supplemen a y Ma e ial, Table S4], we
i s ex ac ed esul s o he 12 o he 21 p e iously epo ed
HR-associa ed SNVs co e ed on he Exome Chip (12). In con as
o p e ious esul s o Eu opeans, only wo known HR-SNVs
showed e idence o associa ion (P<0.05), a he GJA1 and MYH6
loci, in he AA popula ion only. This is likely due o a lack o
powe om he smalle non-Eu opean sample sizes, conside -
ing he powe was calcula ed o be only 48, 11.7 and 8.5% o AA,
HIS and CH, espec i ely. Conco dance in he di ec ion o e ec s
compa ed wi h Eu opeans was only signi ican o AA, wi h 92,
64 and 50% conco dance, co esponding o P- alues o
2.9 10
3
, 0.16 and 0.23 om binomial es s o AA, HIS and CH,
Figu e 2. Manha an plo o he RR-in e al disco e y me a-analysis in Eu opean indi iduals. The Manha an plo displays he esul s om he disco e y me a-analy-
sis o RR-in e als om N¼104,452 indi iduals o Eu opean ances y ( om 30 coho s). On he X axis, P- alues a e exp essed as log
10
(P) a e plo ed acco ding o phys-
ical genomic loca ions by ch omosome. The Y-axis is unca ed o log
10
(P)¼20 wi h any a ian s wi h P <110
20
displayed on he log
10
(P)¼20 line. The nine
no el a ian s alida ed om he combined me a-analysis wi h UK Biobank da a a e ep esen ed by squa es. Va ian s in linkage disequilib ium (LD;
2
>0.8) wi h pub-
lished GWAS a ian s a e highligh ed wi h black ci cles (12). New seconda y a ian s alida ed in ou analysis a e indica ed as iangles. Locus names o he no el loci
co espond o he nea es anno a ed gene, wi h 5p13.3 deno ing an in e genic a ian . The dashed line indica es a P- alue h eshold o 1 10
5
, co esponding o he
lookup signi icance h eshold and he con inuous line indica es a P- alue h eshold o 2 10
7
, co esponding o exome-wide signi icance.
Table 1. Hea a e-associa ed loci iden i ied om Exome Chip analysis
SNV Locus Ch :Pos EA EAF Ndisco e y BETA-RR (SE) Pdisco e y BETA-HR (SE) P eplica ion Pcombined
(A) Fi e un epo ed no el loci
s17853159
a
TESK2 1:45810865 A 0.07 104 452 6.03 (1.20) 5.02 10
7
0.31 (0.08) 9.55 10
5
4.09 10
10
s3087866
a
DALRD3 3:49054692 T 0.25 104 452 3.29 (0.72) 4.92 10
6
0.31 (0.05) 7.06 10
10
2.09 10
14
s1635852 JAZF1 7:28189411 C 0.50 104 452 2.96 (0.62) 2.04 10
6
0.15 (0.04) 4.10 10
4
6.97 10
9
s10857472
a
C10o 71 10:50534599 A 0.45 104 452 2.97 (0.63) 2.11 10
6
0.16 (0.04) 1.49 10
4
2.21 10
9
s3793706
a,b
SEC31B 10:102269085 A 0.22 104 452 3.52 (0.75) 2.54 10
6
0.19 (0.05) 2.06 10
4
3.72 10
9
(B) Fou loci alida ed in ou s udy and also ecen ly published in he UK Biobank s udy
s709209
a
RNF207 1:6278414 G 0.35 104 452 3.30 (0.66) 4.94 10
7
0.27 (0.04) 2.14 10
9
5.44 10
15
s6795970
a
SCN10A 3:38766675 A 0.40 104 452 2.97 (0.64) 3.10 10
6
0.24 (0.04) 1.81 10
8
2.73 10
13
s4282331 5p13.3 5:30881510 G 0.42 104 452 3.56 (0.63) 2.03 10
8
0.26 (0.04) 2.97 10
9
3.34 10
16
s12004
a
KDELR3 22:38877461 G 0.30 104 452 3.30 (0.68) 1.24 10
6
0.31 (0.05) 4.92 10
11
4.04 10
16
Due o he in e se ela ionship be ween R-R in e al and HR he opposi e be a di ec ions do ela e o conco dan di ec ions o e ec be ween disco e y and eplica ion.
SNV, single-nucleo ide a ian ; Ch :Pos, Ch omosome:Posi ion based on HG build 19; EA, e ec allele; EAF, e ec allele equency om he disco e y da a; BETA-RR,
be a e ec es ima e o RR-in e al (milliseconds) aken om he ExomeRR disco e y da a; SE, s anda d e o o he e ec es ima e; N, sample size analysed pe a ian
(p o ided o geno yped disco e y da a only, as eplica ion da a was impu ed so N¼maximum N o all a ian s); BETA-HR, be a e ec o hea a e (in bea s pe mi-
nu e) aken om he UK Biobank eplica ion da a; P,P- alue om ei he he disco e y me a-analysis, he eplica ion da a, o he combined me a-analysis o disco e y
and eplica ion da a. Locus name indica es he nea es gene o he HR-associa ed SNV.
a
Indica es ha he lead o a p oxy SNV (
2
>0.8) is a non-synonymous SNV.
b
Indica es i he lead SNV is p edic ed o be damaging. Mapping o mo e han 500 kb om ei he side o a p e iously epo ed HR-associa ed SNV. A no el locus is a
genomic egion wi h no SNVs in LD (
2
<0.2) wi h HR-associa ed SNVs.
2352 |Human Molecula Gene ics, 2017, Vol. 26, No. 12
espec i ely. The lack o suppo o p e ious indings om he
unde -powe ed non-Eu opean da a led us o es ic ou p i-
ma y disco e y me a-analysis o Eu opeans only.
We also pe o med a look-up o he nine alida ed SNV asso-
cia ions in he non-Eu opean samples. Due o he lack o powe ,
and di e en allele equencies compa ed wi h Eu opeans,
none o he SNVs had esul s wi h P<0.05 wi hin any ances y
(Supplemen a y Ma e ial, Fig. S6), and he e was li le conco d-
ance in e ec di ec ions: 56% and P¼0.246 o AA; 33% and
P¼0.164 o HIS and CH.
Gene-based es s
Gene-based es ing was pe o med o iden i y genes which may
ha e mul iple a e a ian associa ions. None o he gene-based
es esul s was signi ican , a e excluding he single mos sig-
ni ican low- equency a ian om he es s (Supplemen a y
Ma e ial, Table S5).
Look-up o UK Biobank HR-SNVs
Since comple ion o ou me a-analysis o Exome Chip geno-
ypes, a genome-wide scan o HR has been comple ed in UK
Biobank (17). This s udy published 46 new HR loci. Fou o hese
no el loci we e simul aneously disco e ed in ou analyses
(RNF207,SCN10A,5p13.3,KDELR3:Table 1B). Among he 42 e-
maining UK Biobank loci, only i e o he lead SNVs we e co -
e ed on he Exome Chip a
2
0.8. Resul s om ou exome RR
Eu opean-ances y me a-analyses show suppo o all i e o
hese loci (P<0.01; Bon e oni-adjus ed signi icance o i e
es s; Supplemen a y Ma e ial, Table S6).
HR loci and associa ion wi h o he ai s
To p o ide insigh s in o possible sha ed ae iologies o mechan-
isms o disease, we assessed associa ion o ou i e un epo ed
no el HR-SNVs (and hei p oxies,
2
0.8) wi h o he ai s.
Genome-wide signi ican pheno ype–geno ype associa ions
we e obse ed o h ee no el loci (Supplemen a y Ma e ial,
Table S7). The SNV a he DLRD3 locus was associa ed wi h age
o mena che. The SNV a he JAZF1 locus was highly pleio opic,
as shown by associa ions wi h se e al au oimmune diso de s
(sys emic lupus e y hema osus, C ohn’s disease and selec i e
immunoglobulin A de iciency), heigh , ype 2 diabe es and
JAZF1 ansc ip le els in adipose issue. The SNV a he SEC31B
locus was associa ed wi h plasma palmi oleic acid le els and
di e en ial exon exp ession o SEC31B.
Func ional anno a ion o no el HR-SNVs and candida e
genes
Fou o he i e un epo ed no el HR-SNVs o hei p oxies
(
2
>0.8) a e non-synonymous SNVs in TESK2,DALRD3,C10o 71
and SEC31B (Table 1A). The non-synonymous SNV in SEC31B
( s2295774, c.1096T>G, p.Se 332Ala) is in a conse ed egion o
he p o ein, and is p edic ed o be damaging using h ee di e -
en algo i hms in ANNOVAR (19). We also in es iga ed whe he
he no el HR-associa ed SNVs o hei p oxies (
2
>0.8) we e
associa ed wi h changes in exp ession le els o nea by genes
(i.e. as exp ession quan i a i e ai loci, o eQTLs) in he
Geno ype-Tissue Exp ession da abase (GTEx) da ase (20). We
obse ed a signi ican eQTL associa ion a one no el HR locus
(Supplemen a y Ma e ial, Table S8). Speci ically, he HR
inc easing allele o he non-synonymous SNV a SEC31B was
associa ed wi h inc eased le els o SEC31B in ibial ne es
(P¼8.08 10
33
), lung (P¼1.22 10
23
), a ial appendage issue
(P¼4.56 10
11
) and he le en icle (P¼4.0 10
9
), issues
which may be ega ded as physiologically ele an o HR.
We also obse ed HR loci o be signi ican ly en iched o
DNase I hype sensi i e si es (DHSs; Fig. 3). We e alua ed egions
con aining he i e un epo ed no el HR loci and i e independ-
en seconda y a ian s a p e iously epo ed HR loci (12) o-
ge he wi h all 67 published HR-associa ed SNVs [21 loci
epo ed om he o iginal GWAS (12) plus 46 loci ecen ly pub-
lished om UK Biobank (17)]. Highes en ichmen o DHSs in
HR loci occu ed wi hin egions ha a e ansc ip ionally ac i e
in e al hea issue and e al lung, as epo ed in he UK
Biobank s udy. Mo eo e , o he i s ime we ound signi ican
en ichmen o DHSs in human neu onal p ogeni o cells
(de i ed om emb yonic s em cells) and e al muscle samples,
wi h he inclusion o ou no el loci.
Pa hway analyses
We used Ingenui y pa hway analyses o de e mine whe he
he e was any inc eased en ichmen in HR-associa ed pa hways
wi h he con ibu ion o ou i e newly iden i ied loci. We iden-
i ied 16 signi ican ly en iched pa hways a P<110
4
. Mos o
hese pa hways a e ela ed o he ca dio ascula sys em and in-
ol e, o example, sup a en icula a hy hmias, dila ed ca -
diomyopa hy and HR (Supplemen a y Ma e ial, Table S9).
Coding a ian s a HR loci
The Exome Chip p o ides a unique oppo uni y o sea ch o
coding a ian s wi hin known HR loci. Al hough GWAS analyses
ypically iden i y in on o in e genic a ian s, Exome Chip ana-
lysis may iden i y HR-associa ed coding a ian s, which would
poin o candida e causal genes. We conside ed all 67 published
HR loci [21 p e iously epo ed GWAS loci (12) plus 46 ecen ly
published loci om UK Biobank (17)] and ex ac ed all SNVs in
high LD wi h he lead a ian s (
2
0.8), agging he same asso-
cia ion signal, es ic ed o a ian s co e ed on he Exome Chip.
We u he il e ed a ian s o ob ain SNVs ha eached exome-
wide signi icance o associa ions wi h RR-in e al in ou p i-
ma y disco e y me a-analysis, o ensu e ha a ian s ha e a
highly signi ican associa ion wi h he ai . Coding SNVs we e
iden i ied, using he CHARGE Exome Chip anno a ion ile.
We only obse ed wo such coding a ian s in wo epo ed
loci: CCDC141 and KIAA1755. The published CCDC141 coding
a ian was p e iously anno a ed as being non-synonymous
(12), and is p edic ed o be damaging in ou anno a ion
( s17362588; p.A g935T p). The coding SNV a KIAA1755 is he
bes p oxy (
2
1) o he published non-synonymous SNV
( s6127471) co e ed on he Exome Chip (Supplemen a y
Ma e ial, Table S1). The o iginal GWAS (12) had epo ed his
signal as non-synonymous. The e o e, ou Exome Chip analyses
do no e eal any new e idence o likely causal coding a ian s
a well-es ablished HR loci.
Regula o y a ian s a HR loci
Ou analyses o coding a ian s a all known HR loci indica ed
ha he majo i y o HR-associa ed SNVs and he a ian s in
high LD wi h hem a e non-coding. We hus in es iga ed which
a ian s could ha e a causal e ec h ough egula o y
2353Human Molecula Gene ics, 2017, Vol. 26, No. 12 |
ch oma in in e ac ions, such as p omo e –enhance con ac s.
We conside ed all 67 published HR loci [21 p e iously epo ed
GWAS loci (12) plus 46 ecen ly published loci om UK Biobank
(17)], and he i e no el loci epo ed he e. We ound a ian s
ha po en ially a ec enhance unc ion using RegulomeDB
(21) and ound genes whose p omo e egions o m signi ican
ch oma in in e ac ion wi h hem om igh en icle Hi-C da a
(22). We ound 64 po en ial a ge genes in 49 HR loci (4 new loci,
18 loci om he GWAS and 27 loci om he UK Biobank s udy;
Supplemen a y Ma e ial, Table S10). Including hese long- ange
in e ac o s in he candida e causal genes lis inc eased he sig-
ni icance o en ichmen o many HR- ela ed e ms, such as a -
hy hmia and ca diac ib illa ion in ou Ingenui yV
RPa hway
Analysis (IPAV
R; Supplemen a y Ma e ial, Table S11).
Fo newly iden i ied loci, he TESK2 p omo e had a long-
ange in e ac ion wi h he SNVs wi h highes egula o y
po en ial in he locus, unde lining i as a candida e. LOC441204,
a gene o unknown unc ion was ound o in e ac wi h he
JAZF1 locus. A he SEC31B locus, he e we e in e ac ions wi h
wo genes, SCD and SLF2. A he C10o 71 locus, MAPK8 showed
he mos signi ican in e ac ion.
In he 21 loci om he p e iously published GWAS (12), we
iden i ied signi ican ch oma in con ac s o he egula o y
SNVs o 18 loci. We ound CALCRL,TTN,HTR2B,PLD1 and CHRM2
as s onges in e ac o s a he TFPI,CCDC141,B3GNT7,FNDC3B
and CHRM2 loci, espec i ely, ou o hese only CALCRL is in LD
(
2
>0.8) wi h he lead SNV. The p e ious s udy (12) unc ionally
es ed 31 candida e genes, hey ound 20 o hem o ha e an HR
pheno ype in ei he D osophila melanogas e o Danio e io expe i-
men s. All i e o he s onges in e ac o genes we e amongs
he 20 genes wi h an HR pheno ype.
Finally, we ound 41 po en ial causal genes ha ha e no
been implica ed by p e ious GWASs. A ew o hese genes ha e
a ca diac unc ion, including RAPGEF4 (18) and PIM1 (23),
whe eas some a e in ol ed in neu onal de elopmen and unc-
ion, e.g. PBX3,NRNX3. These candida es open up new a enues
ha may aid ou unde s anding o HR biology.
Discussion
Ou me a-analysis o Exome Chip geno ypes yielded i e un e-
po ed no el HR loci, and one un epo ed independen new sec-
onda y signal, which was a low- equency non-synonymous
SNV a he p e iously epo ed KIAA1755 locus.Ou da a
s ongly suppo ed he associa ion o SNVs a 11 o he 12 p e i-
ously epo ed GWAS loci ha we e co e ed on he Exome Chip.
All lead SNVs a all alida ed no el loci a e common (MAF 5%)
and ha e simila e ec sizes, which a e smalle han he e ec
sizes o he majo i y o p e iously epo ed SNVs
(Supplemen a y Ma e ial, Fig. S7). Ou s udy did no yield any
a e SNV associa ions wi h HR, indica ing ha much la ge
sample sizes will be equi ed in u u e s udies o ha e su icien
powe o de ec e ec s o any a e a ian s and assess hei con-
ibu ions o HR he i abili y.
The same obse a ion o he need o la ge sample sizes
applies o he analysis o HR loci iden i ied wi hin Eu opeans in
o he ances ies, whe e he lack o signi icance and conco d-
ance in he esul s om non-Eu opean popula ions is mos
likely due o a lack o powe , as well as di e ences in he allele
equencies and LD pa e ns be ween Eu opeans and non-
Eu opeans. As he non-Eu opean samples we e much smalle ,
we did no pe o m a comp ehensi e compa ison ac oss popu-
la ions o a obus ans-e hnic me a-analysis.
Anno a ion o no el HR-SNVs o hei close p oxies, eQTL
analyses and long- ange ch oma in in e ac ions in hea issue
e eal new po en ial causal candida e HR genes
(Supplemen a y Ma e ial, Tables S10 and S12). A he SEC31B
locus he e is a p edic ed damaging non-synonymous a ian in
SEC31B, and SNVs a his locus a e also signi ican ly associa ed
wi h SEC31B exp ession le els. Al hough i s p ecise unc ion is
unknown, he SEC31B gene encodes SEC31 homolog B, a COPII
coa complex componen . SEC31B has been p oposed o unc-
ion in esicle budding, and ca go expo om he endoplasmic
e iculum (24). The gene is ubiqui ously exp essed a low le els,
bu he e a e highe le els o exp ession in he ce ebellum.
The e a e 13 ansc ip s, and hus se e al p edic ed SEC31B p o-
eins. The majo iso o m is 129 kDa, bu he HR-associa ed non-
synonymous SNV maps o all SEC31B ansc ip s. The e a e no
exis ing mouse models, and he p edic ed p o ein does no di -
ec ly in e ac wi h o he p o eins o pa hways cu en ly
Figu e 3. En ichmen o HR-SNVs in DNase I hype sensi i e si es o 299 issue
samples. The igh panel shows he en ichmen o he combined known and
no el (all) HR-SNVs in DNase I hype sensi i i y si es o 212 Roadmap Epigenome
issue samples ( hose wi h posi i e Z-sco es). En ichmen is exp essed as a
Z-sco e compa ed wi h he dis ibu ion o 1000 ma ched backg ound SNV se s.
Signi ican en ichmen s a e shown in ed (Z-sco e 2.58, alse disco e y a e
(FDR) <1.5%), en ichmen s below his h eshold a e shown in blue. The le
panel shows he en ichmen di e ence (DZsco e¼Zsco e
all
Zsco e
known
) o
hose issue samples in which we ound signi ican en ichmen using all SNPs
and ha u he show a posi i e change using all SNVs compa ed wi h only
known SNVs, wi h inc eased en ichmen hence due o he no el loci iden i ied.
2354 |Human Molecula Gene ics, 2017, Vol. 26, No. 12
Danish Pha maceu ical Associa ion, he Augus inus
Founda ion, he Ib Hen iksen Founda ion, he Becke
Founda ion, and he Danish Diabe es Associa ion.
KORA: The KORA s udy was ini ia ed and inanced by he
Helmhol z Zen um Mu¨ nchen Ge man Resea ch Cen e o
En i onmen al Heal h, which is unded by he Ge man Fede al
Minis y o Educa ion and Resea ch (BMBF) and by he S a e o
Ba a ia. Fu he mo e, KORA esea ch was suppo ed wi hin he
Munich Cen e o Heal h Sciences (MC-Heal h), Ludwig-
Maximilians-Uni e si €
a , as pa o LMUinno a i .
Li eLines: The Li eLines Coho S udy, and gene a ion and man-
agemen o GWAS geno ype da a o he Li eLines Coho S udy
is suppo ed by he Ne he lands O ganiza ion o Scien i ic
Resea ch NWO (g an 175.010.2007.006), he Economic S uc u e
Enhancing Fund (FES) o he Du ch go e nmen , he Minis y o
Economic A ai s, he Minis y o Educa ion, Cul u e and
Science, he Minis y o Heal h, Wel a e and Spo s, he
No he n Ne he lands Collabo a ion o P o inces (SNN), he
P o ince o G oningen, Uni e si y Medical Cen e G oningen,
he Uni e si y o G oningen, Du ch Kidney Founda ion and
Du ch Diabe es Resea ch Founda ion.
MESA: This esea ch was suppo ed by he Mul i-E hnic S udy
o A he oscle osis (MESA) con ac s HHSN2682015000031, N01-
HC-95159, N01-HC-95160, N01-HC-95161, N01- HC-95162, N01-
HC-95163, N01-HC-95164, N01-HC-95165, N01-HC-95166, N01-
HC- 95167, N01-HC-95168, N01-HC-95169 and by g an s UL1-TR-
000040, UL1-TR-001079, and UL1-RR-025005 om Na ional
Cen e Resea ch Resou ces (NCRR). Funding o MESA Family
was p o ided by Na ional Ins i u es o Heal h g an s R01-HL-
071205, R01-HL- 071051, R01-HL-071250, R01-HL-071251, R01-HL-
071252, R01-HL-071258, and R01-HL071259, and by UL1-RR-
025005 and UL1RR033176 om NCRR. Funding o MESA SHARe
geno yping was p o ided by NHLBI Con ac N02-HL-6-4278.
The p o ision o geno yping da a was suppo ed in pa by he
Na ional Cen e o Ad ancing T ansla ional Sciences, CTSI
g an UL1TR000124, and he Na ional Ins i u e o Diabe es and
Diges i e and Kidney Disease Diabe es Resea ch Cen e (DRC)
g an DK063491 o he Sou he n Cali o nia Diabe es
Endoc inology Resea ch Cen e .
NEO: The NEO s udy is suppo ed by he pa icipa ing
Depa men s, he Di ision and he Boa d o Di ec o s o he
Leiden Uni e si y Medical Cen e , and by he Leiden Uni e si y,
Resea ch P o ile A ea Vascula and Regene a i e Medicine.
Dennis Mook-Kanamo i is suppo ed by Du ch Science
O ganiza ion (ZonMW-VENI G an 916.14.023).
PROSPER: The PROSPER s udy was suppo ed by an in es iga o
ini ia ed g an ob ained om B is ol-Mye s Squibb. P o . D . J.
W. Jukema is an Es ablished Clinical In es iga o o he
Ne he lands Hea Founda ion (g an 2001 D 032). Suppo o
geno yping was p o ided by he se en h amewo k p og am o
he Eu opean commission (g an 223004) and by he
Ne he lands Genomics Ini ia i e (Ne he lands Conso ium o
Heal hy Aging g an 050-060-810).
RS: The Ro e dam S udy is unded by E asmus Medical Cen e
and E asmus Uni e si y, Ro e dam, Ne he lands O ganiza ion
o he Heal h Resea ch and De elopmen (ZonMw), he
Resea ch Ins i u e o Diseases in he Elde ly (RIDE), he
Minis y o Educa ion, Cul u e and Science, he Minis y o
Heal h, Wel a e and Spo s, he Eu opean Commission (DG XII),
and he Municipali y o Ro e dam. The Exome Chip a ay da a
se was unded by he Gene ic Labo a o y o he Depa men o
In e nal Medicine, E asmus MC, om he Ne he lands
Genomics Ini ia i e (NGI)/Ne he lands O ganisa ion o
Scien i ic Resea ch (NWO)-sponso ed Ne he lands Conso ium
o Heal hy Aging (NCHA; p ojec n . 050-060- 810); he
Ne he lands O ganiza ion o Scien i ic Resea ch (NWO; p ojec
numbe 184021007) and by he Rainbow P ojec (RP10;
Ne he lands Exome Chip P ojec ) o he Biobanking and
Biomolecula Resea ch In as uc u e Ne he lands (BBMRINL;
www.bbm i.nl).
Sa diNIA: This esea ch was suppo ed by Na ional Human
Genome Resea ch Ins i u e g an s HG005581, HG005552,
HG006513, HG007022 and HG007089; by Na ional Hea , Lung,
and Blood Ins i u e g an HL117626; by he In amu al Resea ch
P og am o he US Na ional Ins i u es o Heal h, Na ional
Ins i u e on Aging, con ac s N01-AG-1-2109 and
HHSN271201100005C; by Sa dinian Au onomous Region (L.R. 7/
2009) g an cRP3-154
SHIP: SHIP (S udy o Heal h in Pome ania) and SHIP-TREND
bo h ep esen popula ionbased s udies. SHIP is suppo ed by
he Ge man Fede al Minis y o Educa ion and Resea ch
(Bundesminis e ium u¨ Bildung und Fo schung (BMBF); g an s
01ZZ9603, 01ZZ0103, and 01ZZ0403) and he Ge man Resea ch
Founda ion (Deu sche Fo schungsgemeinscha (DFG); g an GR
1912/5-1). SHIP and SHIP-TREND a e pa o he Communi y
Medicine Resea ch ne (CMR) o he E ns -Mo i z-A nd
Uni e si y G ei swald (EMAU) which is unded by he BMBF as
well as he Minis y o Educa ion, Science and Cul u e and he
Minis y o Labo , Equal Oppo uni ies, and Social A ai s o he
Fede al S a e o Mecklenbu g-Wes Pome ania. The CMR
encompasses se e al esea ch p ojec s ha sha e da a om
SHIP. The EMAU is a membe o he Cen e o Knowledge
In e change (CKI) p og am o he Siemens AG. SNP yping o
SHIP and SHIP-TREND using he Illumina In inium
HumanExome BeadChip ( e sion 1.0) was suppo ed by he
Fede al Minis y o Educa ion and Resea ch (BMBF) g an
03Z1CN22.
TwinsUK: This wo k was unded by a g an om he B i ish
Hea Founda ion (PG/12/38/29615). The TwinsUK s udy was
unded by he Wellcome T us ; Eu opean 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
UHP: The U ech Heal h P ojec ecei ed g an s om he
Minis y o Heal h, Wel a e and Spo s (VWS), he Uni e si y o
U ech , he P o ince o U ech , he Du ch O ganisa ion o
Ca e. Resea ch, he Uni e si y Medical Cen e o U ech , and
he Du ch College o Heal hca e Insu ance Companies. The
Exome Chip da a we e gene a ed in a esea ch p ojec ha was
inancially suppo ed by Biobanking and Biomolecula esou ces
Resea ch In as uc u e (BBMRI-NL, a Resea ch In as uc u e
inanced by he Du ch go e nmen (NWO 184.021.007).
WHI: The WHI p og am is unded by he Na ional Hea , Lung,
and Blood Ins i u e, Na ional Ins i u es o Heal h, U.S.
Depa men o Heal h and Human Se ices h ough con ac s
HHSN268201600018C, HHSN268201600001C, HHSN268201600002C,
HHSN268201600003C, and HHSN268201600004C.
YFS: The Young Finns S udy has been inancially suppo ed by
he Academy o Finland: g an s 286284, 134309 (Eye), 126925,
121584, 124282, 129378 (Sal e), 117787 (Gendi), and 41071 (Skidi);
he Social Insu ance Ins i u ion o Finland; Kuopio, Tampe e
and Tu ku Uni e si y Hospi al Medical Funds (g an X51001);
Juho Vainio Founda ion; Paa o Nu mi Founda ion; Finnish
Founda ion o Ca dio ascula Resea ch; Finnish Cul u al
Founda ion; Tampe e Tube culosis Founda ion; Emil Aal onen
Founda ion; Y jo¨ Jahnsson Founda ion; Signe and Ane
2361Human Molecula Gene ics, 2017, Vol. 26, No. 12 |
Gyllenbe g Founda ion; and Diabe es Resea ch Founda ion o
Finnish Diabe es Associa ion. Funding o pay he Open Access
publica ion cha ges o his a icle was p o ided by he Medical
Resea ch Council G an Numbe : MR/N025083/1.
URLs
h p://www.g expo al.o g; da e las accessed Janua y 10, 2017.
h p://www.phenoscanne .medschl.cam.ac.uk; da e las accessed
Janua y 11, 2017.
h p://www.ingenui y.com
h p://www.geneca ds.o g; da e las accessed Ma ch 10, 2017.
h p://omim.o g; da e las accessed Augus 26, 2016.
h p://www.p o eina las.o g/; da e las accessed Ma ch 10, 2017.
h p://s ing-db.o g/; da e las accessed No embe 5, 2016.
h p://www.unip o .o g; da e las accessed Ma ch 10, 2017.
h p://www.in e na ionalgenome.o g/ o ge-analysis/; da e las
accessed No embe 29, 2016.
Re e ences
1. Aladin, A.I., Whel on, S.P., Al-Mallah, M.H., Blaha, M.J.,
Ke eyian, S.J., Ju aschek, S.P., Rubin, J., B awne , C.A. and
Michos, E.D. (2014) Rela ion o es ing hea a e o isk o
all-cause mo ali y by gende a e conside ing exe cise cap-
aci y ( he Hen y Fo d exe cise es ing p ojec ). Am. J. Ca diol.,
114, 1701–1706.
2. Ca lson, N., Dixen, U., Ma o , J.L., Jensen, M.T. and Jensen,
G.B. (2014) P edic i e alue o casual ECG-based es ing hea
a e compa ed wi h es ing hea a e ob ained om Hol e
eco ding. Scand. J. Clin. Lab. In es ., 74, 163–169.
3. Fox, K., Bousse , M.G., Ama enco, P., Chamo o, A., Fishe ,
M., Fo d, I., Henne ici, M.G., Ma le, H.P. and Ro hwell, P.M.
(2013) Hea a e is a p ognos ic isk ac o o myoca dial in-
a c ion: a pos hoc analysis in he PERFORM (P e en ion o
ce eb o ascula and ca dio ascula E en s o ischemic o i-
gin wi h eRu oban in pa ien s wi h a his o y oF ischemic
s Oke o Ransien ischeMic a ack) s udy popula ion. In . J.
Ca diol., 168, 3500–3505.
4. Woodwa d, M., Webs e , R., Mu akami, Y., Ba zi, F., Lam,
T.H., Fang, X., Suh, I., Ba y, G.D., Huxley, R. and Rodge s, A.
(2014) The associa ion be ween es ing hea a e, ca dio as-
cula disease and mo ali y: e idence om 112,680 men and
women in 12 coho s. Eu . J. P e . Ca diol., 21, 719–726.
5. Jou en, X., Zu eik, M., Desnos, M., Gue o , C. and
Ducime ie e, P. (2001) Res ing hea a e as a p edic i e isk
ac o o sudden dea h in middle-aged men. Ca dio asc.
Res., 50, 373–378.
6. Teodo escu, C., Reinie , K., Uy-E anado, A., Gunson, K., Jui, J.
and Chugh, S.S. (2013) Res ing hea a e and isk o sudden
ca diac dea h in he gene al popula ion: in luence o le
en icula sys olic dys unc ion and hea a e-modula ing
d ugs. Hea Rhy hm,10, 1153–1158.
7. Secca eccia, F., Pannozzo, F., Dima, F., Minop io, A.,
Mendi o, A., Lo Noce, C., Giampaoli, S. and Mala ie
Ca dio ascola i A e oscle o iche Is i u o Supe io e di Sani a
P ojec (2001) Hea a e as a p edic o o mo ali y: he
MATISS p ojec . Am. J. Public Heal h,91, 1258–1263.
8. Fox, K.M. and Fe a i, R. (2011) Hea a e: a o go en link in
co ona y a e y disease? Na . Re . Ca diol., 8, 369–379.
9. Singh, J.P., La son, M.G., O’Donnell, C.J., Tsuji, H., E ans, J.C.
and Le y, D. (1999) He i abili y o hea a e a iabili y: he
F amingham Hea S udy. Ci cula ion,99, 2251–2254.
10. Ma in, L.J., Comuzzie, A.G., Sonnenbe g, G.E., Myklebus , J.,
James, R., Ma ks, J., Blange o, J. and Kissebah, A.H. (2004)
Majo quan i a i e ai locus o es ing hea a e maps o a
egion on ch omosome 4. Hype ension,43, 1146–1151.
11. Wang, B., Liao, C., Zhou, B., Cao, W., L , J., Yu, C., Gao, W. and
Li, L. (2015) Gene ic con ibu ion o he a iance o blood
p essu e and hea a e: a sys ema ic e iew and me a-
eg ession o win s udies. Twin Res. Hum. Gene ., 18, 158–170.
12. den Hoed, M., Eijgelsheim, M., Esko, T., B undel, B.J., Peal,
D.S., E ans, D.M., Nol e, I.M., Seg e, A.V., Holm, H.,
Handsake , R.E. e al. (2013) Iden i ica ion o hea a e-
associa ed loci and hei e ec s on ca diac conduc ion and
hy hm diso de s. Na . Gene ., 45, 621–631.
13. Deo, R., Nalls, M.A., A e y, C.L., Smi h, J.G., E ans, D.S.,
Kelle , M.F., Bu le , A.M., Buxbaum, S.G., Li, G., Miguel
Quib e a, P. e al. (2013) Common gene ic a ia ion nea he
connexin-43 gene is associa ed wi h es ing hea a e in
A ican Ame icans: a genome-wide associa ion s udy o
13,372 pa icipan s. Hea Rhy hm,10, 401–408.
14. Eijgelsheim, M., New on-Cheh, C., So oodehnia, N., de
Bakke , P.I., Mulle , M., Mo ison, A.C., Smi h, A.V., Isaacs, A.,
Sanna, S., Do , M. e al. (2010) Genome-wide associa ion
analysis iden i ies mul iple loci ela ed o es ing hea a e.
Hum. Mol. Gene ., 19, 3885–3894.
15. Holm, H., Gudbja sson, D.F., A na , D.O., Tho lei sson, G.,
Tho gei sson, G., S e ansdo i , H., Gudjonsson, S.A.,
Jonasdo i , A., Ma hiesen, E.B., Njols ad, I. e al. (2010)
Se e al common a ian s modula e hea a e, PR in e al
and QRS du a ion. Na . Gene ., 42, 117–122.
16. Ma oni, F., P eu e , A., Aulchenko, Y.S., F anklin, C.S.,
Isaacs, A., Pichle , I., Wild, S.H., Oos a, B.A., W igh , A.F.,
Campbell, H. e al. (2009) A genome-wide associa ion scan o
RR and QT in e al du a ion in 3 Eu opean gene ically iso-
la ed popula ions: he EUROSPAN p ojec . Ci c. Ca dio asc.
Gene ., 2, 322–328.
17. Eppinga, R.N., Hagemeije , Y., Bu gess, S., Hinds, D.A.,
S e ansson, K., Gudbja sson, D.F., an Veldhuisen, D.J., Mun oe,
P.B., Ve weij, N. and an de Ha s , P. (2016) Iden i ica ion o gen-
omic loci associa ed wi h es ing hea a e and sha ed gene ic
p edic o s wi h all-cause mo ali y. Na . Gene ., 48, 1557–1563.
18. Sugawa a, K., Shibasaki, T., Takahashi, H. and Seino, S.
(2016) S uc u e and unc ional oles o Epac2 (Rapge 4).
Gene,575, 577–583.
19. Wang, K., Li, M. and Hakona son, H. (2010) ANNOVAR: unc-
ional anno a ion o gene ic a ian s om high- h oughpu
sequencing da a. Nucleic Acids Res., 38, e164.
20. GTEx Conso ium (2013) The Geno ype-Tissue Exp ession
(GTEx) p ojec . Na . Gene ., 45, 580–585.
21. Boyle, A.P., Hong, E.L., Ha iha an, M., Cheng, Y., Schaub,
M.A., Kasowski, M., Ka czewski, K.J., Pa k, J., Hi z, B.C., Weng,
S. e al. (2012) Anno a ion o unc ional a ia ion in pe sonal
genomes using RegulomeDB. Genome Res., 22, 1790–1797.
22. Schmi , A.D., Hu, M., Jung, I., Xu, Z., Qiu, Y., Tan, C.L., Li, Y.,
Lin, S., Lin, Y., Ba , C.L. e al. (2016) A compendium o ch o-
ma in con ac maps e eals spa ially ac i e egions in he
human genome. Cell Rep., 17, 2042–2059.
23. Wallne , M., Kolesnik, E., Ablasse , K., Kha aga, M., Wakula,
P., Ljuboje ic, S., Thon-Gu schi, E.M., Sou ij, H., Kapl, M.,
Edmunds, N.J. e al. (2015) Exena ide exe s a PKA-dependen
posi i e ino opic e ec in human a ial myoca dium: GLP-
1R media ed e ec s in human myoca dium. J. Mol. Cell
Ca diol., 89, 365–375.
24. S ankewich, M.C., S abach, P.R. and Mo ow, J.S. (2006)
Human Sec31B: a amily o new mammalian o hologues o
2362 |Human Molecula Gene ics, 2017, Vol. 26, No. 12
yeas Sec31p ha associa e wi h he COPII coa . J. Cell Sci.,
119, 958–969.
25. Dob zyn, P., Dob zyn, A., Miyazaki, M. and N ambi, J.M. (2010)
Loss o s ea oyl-CoA desa u ase 1 escues ca diac unc ion in
obese lep in-de icien mice. J. Lipid Res., 51, 2202–2210.
26. Rosok, O., Pedeu ou , F., Ree, A.H. and Aasheim, H.C. (1999)
Iden i ica ion and cha ac e iza ion o TESK2, a no el mem-
be o he LIMK/TESK amily o p o ein kinases, p edomin-
an ly exp essed in es is. Genomics,61, 44–54.
27. Rose, B.A., Fo ce, T. and Wang, Y. (2010) Mi ogen-ac i a ed
p o ein kinase signaling in he hea : angels e sus demons
in a hea -b eaking ale. Physiol. Re ., 90, 1507–1546.
28. Chen,Q.,Xu,T.,Li,D.,Pan,D.,Wu,P.,Luo,Y.,Ma,Y.andLiu,Y.
(2016) JNK/PI3K/Ak signaling pa hway is in ol ed in myoca -
dial ischemia/ epe usion inju y in diabe ic a s: e ec s o sal-
ianolic acid A in e en ion. Am.J.T ansl.Res., 8, 2534–2548.
29. G inchuk, O.V., Jenja oenpun, P., O lo , Y.L., Zhou, J. and
Kuzne so , V.A. (2010) In eg a i e analysis o he human cis-
an isense gene pai s, miRNAs and hei ansc ip ion egula-
ion pa e ns. Nucleic Acids Res., 38, 534–547.
30. Fukuda, T., Sugi a, S., Ina ome, R. and Yanagi, S. (2010) CAMDI,
a no el dis up ed in schizoph enia 1 (DISC1)-binding p o ein,
is equi ed o adial mig a ion. J. Biol. Chem., 285, 40554–40561.
31. LeWin e , M.M. and G anzie , H.L. (2013) Ti in is a majo
human disease gene. Ci cula ion,127, 938–944.
32. G o e, M.L., Yu, B., Coch an, B.J., Ha i unians, T., Bis, J.C.,
Taylo , K.D., Hansen, M., Bo ecki, I.B., Cupples, L.A., Fo nage,
M. e al. (2013) Bes p ac ices and join calling o he
HumanExome BeadChip: he CHARGE Conso ium. PLoS One,
8, e68095.
33. Wille , C.J., Li, Y. and Abecasis, G.R. (2010) METAL: as and
e icien me a-analysis o genomewide associa ion scans.
Bioin o ma ics,26, 2190–2191.
34. Yang, J., Fe ei a, T., Mo is, A.P., Medland, S.E., Madden,
P.A., Hea h, A.C., Ma in, N.G., Mon gome y, G.W., Weedon,
M.N., Loos, R.J. e al. (2012) Condi ional and join mul iple-
SNP analysis o GWAS summa y s a is ics iden i ies add-
i ional a ian s in luencing complex ai s. Na . Gene ., 44,
369–375, S361–S363.
35. Pu cell, S., Che ny, S.S. and Sham, P.C. (2003) Gene ic Powe
Calcula o : design o linkage and associa ion gene ic map-
ping s udies o complex ai s. Bioin o ma ics,19, 149–150.
36. S aley, J.R., Blackshaw, J., Kama , M.A., Ellis, S., Su end an,
P., Sun, B.B., Paul, D.S., F ei ag, D., Bu gess, S., Danesh, J. e al.
(2016) PhenoScanne : a da abase o human geno ype-
pheno ype associa ions. Bioin o ma ics,32, 3207–3209.
37. Dunham, I., Kulesha, E., Io chko a, V., Mo ganella, S. and
Bi ney, E. (2014) FORGE: a ool o disco e cell speci ic en ich-
men s o GWAS associa ed SNPs in egula o y egions.
bioRxi doi:h p://dx.doi.o g/10.1101/013045.
2363Human Molecula Gene ics, 2017, Vol. 26, No. 12 |