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Rinnakkaistallenteet Luonnontieteiden ja metsätieteiden tiedekunta
2021
A Novel Genetic Marker for the C9orf72 Repeat Expansion in the Finnish Population
Rostalski, H
IOS Press
Tieteelliset aikakauslehtiartikkelit þÿ© 2021 IOS Press
All rights reserved
http://dx.doi.org/10.3233/JAD-210599
https://erepo.uef.fi/handle/123456789/26367
Downloaded from University of Eastern Finland's eRepository
1 Article
1
A novel genetic marker for the C9orf72 repeat expansion in the Finnish population 2
Hannah Rostalskia, Ville Korhonenb, Teemu Kuulasmaac, Eino Soljed,e, Johanna Krügerf,g, 3
FinnGen, Karri Kaivolah, Per Kristian Eidei, Jean-Charles Lambertj, Valtteri Julkunend,e, 4
Pentti J. Tienarih, Anne M. Remesf,g, Ville Leinonenb, Mikko Hiltunenc*, Annakaisa 5
Haapasaloa* 6
aA. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, P. O. Box 1627 7
(Neulaniementie 2), 70211 Kuopio, Finland.
8
bNeurocenter, Neurosurgery, Kuopio University Hospital and University of Eastern Finland, 9
Kuopio, Finland.
10
cInstitute of Biomedicine, Yliopistonranta 1E, University of Eastern Finland, 70211 Kuopio, 11
Finland.
12
dInstitute of Clinical Medicine - Neurology, University of Eastern Finland.
13
eNeuro Center, Neurology, Kuopio University Hospital, Kuopio, Finland.
14
fResearch Unit of Clinical Neuroscience, Neurology, University of Oulu, Oulu, Finland.
15
gMedical Research Center (MRC), Oulu University Hospital, Oulu, Finland.
16
hDepartment of Neurology, Helsinki University Hospital and Translational Immunology 17
Program, Biomedicum, University of Helsinki, Helsinki, Finland.
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iOslo University Hospital-Rikshospitalet; and Institute of Clinical Medicine, Faculty of 19
Medicine, University of Oslo, Norway.
20
jUniv. Lille, Inserm, CHU Lille, Institut Pasteur de Lille, U1167-RID-AGE facteurs de risque 21
et déterminants moléculaires des maladies liés au vieillissement, Lille, France.
22
2
*These authors contributed equally to this work 23
Corresponding authors:
24
*Annakaisa Haapasalo, PhD, Adjunct Professor, Research Director 25
Email: annakaisa.haapasalo@uef.fi; Tel: +358403552768 26
and 27
*Mikko Hiltunen, PhD, Professor 28
Email: mikko.hiltunen@uef.fi; Tel: +358403552014 29
30
Running title: A novel C9orf72 repeat expansion-associated SNP 31
32
Disclosure of support for the work:
33
This work was supported by the Academy of Finland, under grant numbers 315459 (AH), 34
315460 (AMR), and 307866 (MH); the Strategic Neuroscience Funding of the University of 35
Eastern Finland; Finnish Brain Foundation (ES), Sigrid Juselius Foundation (ES, MH), 36
Instrumentarium Science Foundation (ES), Orion Research Foundation (ES). This publication 37
is part of a project that has received funding from the European Union’s Horizon 2020 research 38
and innovation programme under the Marie Skłodowska-Curie grant agreement No 740264.
39
40
41
3 Abstract
42
Background: C9orf72 repeat expansion (C9exp) is the most common genetic cause underlying 43
frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). However, 44
detection of the C9exp requires elaborative methods. Objective: Identification of C9exp carriers 45
from genotyped cohorts could be facilitated by using single nucleotide polymorphisms (SNPs) 46
as markers for the C9exp. Methods: We elucidated the potential of the previously described 47
Finnish risk haplotype, defined by the SNP rs3849942, to identify potential C9exp carriers 48
among 218,792 Finns using the FinnGen database. The haplotype approach was first tested in 49
an idiopathic normal pressure hydrocephalus (iNPH) patient cohort (European Alzheimer’s 50
Disease DNA BioBank) containing C9exp carriers by comparing intermediate (15-30) and full- 51
length (> 60 repeats) C9exp carriers (n = 41) to C9exp negative patients (< 15 repeats, n = 801).
52
Results: In this analysis, rs3849942 was associated with carriership of C9exp (OR 8.44, 53
p < 2×10-15), while the strongest association was found with rs139185008 (OR 39.4, p < 5×10- 54
18). Unbiased analysis of rs139185008 in FinnGen showed the strongest association with FTLD 55
(OR 4.38, 3×10-15) and motor neuron disease ALS (OR 5.19, 3×10-21). rs139185008 was the 56
top SNP in all diseases (iNPH, FTLD, ALS), and further showed a strong association with ALS 57
in the UK Biobank (p = 9.0×10-8). Conclusion: Our findings suggest that rs139185008 is a 58
useful marker to identify potential C9exp carriers in the genotyped cohorts and biobanks 59
originating from Finland.
60
61
Keywords: Amyotrophic lateral sclerosis; C9orf72; DNA Repeat Expansion; Frontotemporal 62
lobar degeneration; Motor neuron disease; Polymorphism, Single Nucleotide 63
64
4 Introduction
65
Frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) are 66
neurodegenerative disorders sharing genetic and neuropathological similarities [1]. C9orf72 67
hexanucleotide repeat expansion (C9exp), the most common genetic cause of FTLD and ALS 68
[2,3], is exceptionally prevalent in Finnish FTLD and ALS patients [4,5]. Previous studies have 69
suggested that more than 30 units of the C9exp are pathogenic [3,6]. Recently, the C9exp was 70
shown to be an important genetic etiology for idiopathic normal pressure hydrocephalus (iNPH) 71
[7], which is the most common form of hydrocephalus and characterized by progressive gait 72
impairment, cognitive decline, and loss of bladder control [8]. Because it is not possible to 73
sequence the expanded region using whole genome-sequencing, the presence and estimated 74
length of the C9exp can only be determined using repeat-primed PCR, Southern blotting or long- 75
read sequencing [9,10]. The discovery of specific single nucleotide polymorphisms (SNPs) and 76
groups of SNPs (haplotypes) associating with the C9exp would enable identification of potential 77
C9exp carriers from large genotyped cohorts from which the C9exp cannot be detected using 78
current methods. Previously, a Finnish risk haplotype of 42 single nucleotide polymorphisms 79
(SNP) was reported to associate with ALS in Finland [11]. Moreover, another risk haplotype of 80
20 SNPs has been shown to associate with FTLD, ALS, and the C9exp in other European and 81
U.S. cohorts [12]. Here, we aimed to identify a SNP that could be used as a genetic marker to 82
identify C9exp carriers in Finnish cohorts. Our findings showed that the variant rs139185008 83
distinguishes C9exp carriers from non-carriers in the European Alzheimer’s Disease DNA 84
BioBank (EADB) and associates with the clinical diagnoses of FTLD and ALS in the large 85
population-based FinnGen database and UK Biobank. This suggests that rs139185008 might 86
be a powerful genetic marker for the identification of C9exp carriers in other Finnish cohorts as 87
well.
88
5 Subjects and Methods
89
Cohorts, genotyping and imputations of EADB samples, and clinical endpoints 90
This study includes GWAS data from the EADB and the FinnGen database. EADB data were 91
processed as previously described [13]. Finnish iNPH patients included in the EADB GWAS 92
were diagnosed according to published guidelines and procedures [14,15]. C9exp genotyping 93
was performed using repeat-primed PCR and amplicon length analysis [3]. Previous studies 94
have suggested a pathological threshold of > 30 units [3,6] or > 45 units [16] for the C9exp. 95
However, smaller repeats of < 30 units may also associate with disease [4,6,16,17]. In these 96
studies, the minimum lengths of the C9exp have been identified as 7 [4] and 17 [16,17] repeats 97
on the longer allele. Here, we chose a threshold of 15 repeats to define individuals positive for 98
the C9exp. The iNPH cohort contains 41 C9exp carriers [7 full-length (> 60 repeats) and 34 99
intermediate C9exp carriers (15-30 repeats)] and 801 controls (< 15 repeats). Forty-eight percent 100
of C9exp carriers and controls were male.
101
Detailed information of the FinnGen data is described at https://www.finngen.fi/fi. Genome and 102
clinical data from 218,792 individuals were obtained from FinnGen study data release 5.
103
Clinical diagnoses were derived from the International Statistical Classification of Diseases and 104
Related Health Problems, version 10 (ICD-10) codes in Finnish national hospital registries and 105
cause-of-death registry as part of FinnGen project. UK Biobank data were used for validation 106
of the identified SNPs and haplotypes.
107
Generation of risk haplotypes associating with C9exp 108
Trans-Omics for Precision Medicine (TOPMed) imputed genotype data from EADB 109
consortium was used [13]. Genotypes were phased with Eagle v2.4105 and imputed with 110
Minimac4 v4-1.0.2. Only SNPs having Hardy-Weinberg equilibrium p > 10-5 and imputation 111
quality greater than 0.6 were considered. The imputation quality score for rs139185008 was 112
0.75. The previously published 20-SNP Finnish risk haplotype [12] was used to test for 113
6
association with the C9exp (iNPH cohort) and clinical diagnoses (“motor neuron disease” for 114
ALS, “circumscribed brain atrophy” for FTLD; FinnGen). Additional upstream and 115
downstream SNPs were added to create longer haplotypes that were able to distinguish C9exp
116
carriers better from non-carriers. The SNP selection was conducted based on a side by side 117
inspection of an individual C9exp carrier and non-carrier haplotypes of the phased and imputed 118
most probable genotype data. Minor and major alleles included in the haplotypes are presented 119
in Supplementary Table 1.
120
Analysis of SNP and haplotype association with C9exp and clinical endpoints 121
Both LD-statistics (D’) and case vs. control logistic regression analysis with covariates were 122
conducted on pre-processed imputed genotypes using PLINK software [version 1.9; [18]]. For 123
iNPH cohort, only principal component (PC) 1-2, and for FinnGen PC1-5 were used as 124
covariates. UK Biobank data were extracted through
125
http://big.stats.ox.ac.uk/variant/9:27491942-T-C and http://big.stats.ox.ac.uk/pheno/traits_011.
126
Data Presentation 127
Manhattan and regional association plots were drawn using LocusZoom software (v0.12.0). For 128
LD calculation, European reference population was used. Images were modified using 129
LibreOffice Draw (version: 6.0.2.1). Bar graphs and geographical plot of minor allele 130
frequencies (MAFs) were generated, and Pearson’s Chi-square test on minor vs. major allele 131
counts among Finnish regions was performed using RStudio software (version: 1.1.463) and 132
ggplot2 [19] and geofi packages [20].
133
Data availability 134
Data are available on reasonable request from the corresponding authors. Due to privacy 135
policies, the data are not publicly available.
136
7 Ethics statement
137
All experimental procedures complied with the standards of the Declaration of Helsinki. The 138
Ethics Committee of Hospital District of Northern Savo approved the iNPH study and all 139
patients provided an informed consent. Patients and controls in FinnGen provided informed 140
consent for biobank research, based on the Finnish Biobank Act (https://www.finngen.fi/fi). All 141
DNA samples and data were pseudonymized (iNPH cohort and FinnGen cohort).
142
8 Results
143
C9exp associates with SNPs near MOB3B and C9orf72 genes 144
Based on genotype data obtained from a global screening array, SNP association analysis was 145
performed in a well-characterized iNPH patient cohort, comprising intermediate (15-30) and 146
full-length (> 60 repeats) C9exp carriers (n = 41) who were compared to non-carriers (< 15 147
repeats, n = 801). Except for two SNPs, all significantly C9exp-associated SNPs (p < 5×10-8) 148
were located on chromosome 9 (Figure 1A). Several of these were close or within the MOB 149
kinase activator 3B (MOB3B) or C9orf72 genes, spanning an approx. 94 kb region, and showed 150
a strong linkage disequilibrium (LD) (r2 ≥ 0.8) with the reference SNP rs3849942 (Figure 1B), 151
a previously reported surrogate marker for the chromosome 9p risk haplotype [2,12].
152
Interestingly, rs139185008 (odds ratio, OR = 39.4, 95% CI [17.2-90.5], p = 4.6×10-18), 153
localizing within a recombination-poor region 81 541 bp upstream of the C9exp, showed the 154
strongest single SNP association with C9exp carriership. Also, rs139185008 (MAF 0.016) was 155
in complete LD (D’ = 1.00) with the reference SNP rs3849942 (MAF 0.17), which showed a 156
weaker association with C9exp carriership (OR 8.44, 95% CI [4.99-140.29], p = 2.0×10-15).
157
Importantly, rs139185008 was highly abundant in C9exp carriers (minor allele frequency, MAF 158
for full-length and intermediate carriers = 0.21 and 0.19, respectively), but rare in non-carriers 159
(< 15 repeats, MAF = 0.008). Several C9exp-associated haplotypes were significantly 160
overrepresented in C9exp carriers as compared to non-carriers in the iNPH cohort (Table 1).
161
rs139185008 was part of the haplotypes 2, 5, 6, 8 and 10 showing the most prominent risk 162
effects (OR > 42.0). Moreover, as compared to the previously reported 20-SNP Finnish risk 163
haplotype, including e.g., rs868856, rs7046653, rs2814707, rs3849942, and rs774359 [12]
164
(Figure 1B), the inclusion of rs139185008 to haplotypes (“haplo”) 2, 5, 6, 8 and 10 markedly 165
improved the specificity to identify C9exp carriers from non-carriers in the iNPH cohort (Table 166
1, Supplementary Table 1). E.g., the OR for haplotype 2 (OR = 11.33, 95% CI [6.38-20.14], 167
9
p = 1.28×10-16) substantially increased after the inclusion of rs139185008 (OR = 42.74, 95%
168
CI [18.35-99.53], p = 3.16×10-18).
169
rs139185008 strongly associates with FTLD and ALS in FinnGen 170
Next, we unbiasedly examined in the FinnGen database which clinical diagnoses associate with 171
the SNPs and haplotypes identified in the iNPH cohort. The FinnGen database contains 172
comprehensive genome-wide genotype data and life-long medical history from >200,000 Finns.
173
However, FinnGen does not include genetic data on complex genomic alterations, such as 174
C9exp. rs139185008 and haplotypes 2, 5, 6, 8, and 10 containing the minor allele of rs139185008 175
strongly associated with ALS and FTLD (Table 1). In comparison, the previously reported 20- 176
SNP Finnish risk haplotype [12] showed a weaker association with ALS and FTLD (haplo 2, 177
Table 1). In FinnGen, rs139185008 was the top SNP that associated with ALS and FTLD, 178
confirming the result obtained in the iNPH GWAS (Figure 1B). Importantly, rs139185008 also 179
significantly associated with ALS in UK Biobank (p = 9.0×10-8), but it was not among the top 180
SNPs associated with ALS (beta value = -0.4; p values ≤ 1.2×10-18; Supplementary Figure 1).
181
rs139185008 is regionally enriched to South-Eastern Finland 182
Finally, we used FinnGen data to calculate the MAF of the rs139185008 according to the region 183
of birth in Finland (Figure 2). Geographically, the rs139185008 minor allele showed the highest 184
prevalence in Southern Savonia (MAF = 0.025) and the lowest in Ostrobothnia (MAF = 0.008) 185
(Figure 2). Pearson’s Chi-square test of the frequency of rs139185008 minor allele revealed 186
significant differences in the geographic distribution of rs139185008 in Finland (p < 2.2×10-16, 187
X2 = 282.43, df = 18).
188
Discussion 189
We report that rs139185008 strongly associates with C9exp in a cohort of iNPH cases, suggesting 190
surrogate marker potential for identifying C9exp carriers in large population-based cohorts and 191
10
biobank databases. rs139185008 indicated stronger association with FTLD and ALS clinical 192
diagnoses in FinnGen (OR 4.4 and 5.2, respectively) as compared to the previously reported 193
C9exp proxy marker rs3849942 (OR 1.2 and 1.6 respectively). The top SNPs differed in FinnGen 194
and UK Biobank, which indicates that there are differences in the C9exp haplotype structures 195
among European populations. In Finland, the frequency of rs139185008 minor allele was 196
highest in South-Eastern Finland, and lowest in the west-coastal Ostrobothnia, which represent 197
genetically different geographical regions. The regional distribution of rs139185008 in Finland 198
is consistent with the most enriched areas of haplotypes of Finnish Heritage Diseases in regard 199
of the low enrichment in the west coast area and high enrichment in Savonia regions [21], a 200
phenomenon traceable back to the population migration history within Finland and the resulting 201
genetic isolation due to bottleneck events and founder effects [22]. However, rs139185008 is 202
also highly prevalent in Helsinki and surrounding areas and showed a link to ALS in the UK 203
Biobank, consisting of a more heterogeneous population. In this context, however, it should be 204
emphasized that the beta-value provided by UK Biobank for rs139185008 was negative, 205
indicating an odds ratio below one for this SNP. Importantly, similar results (negative beta- 206
values) were also observed with some other SNPs significantly associated with ALS in the UK 207
Biobank in the MOB3B/C9orf72 region. Thus, further investigations on the prevalence of 208
rs139185008 and its association with C9exp-linked diseases in other populations and cohorts are 209
warranted in the future to evaluate its importance beyond Finland and the UK Biobank.
210
Collectively, the present data suggest that specific haplotypes containing rs139185008 are 211
useful proxy markers to identify potential C9exp carriers. Since gene-based therapies are 212
emerging in C9exp-linked diseases, rs139185008 may be utilized in the identification of 213
potential C9exp carriers already at an early phase from biobanks and population cohorts for 214
confirmatory C9exp genotyping and subsequent clinical trials in the future.
215
Acknowledgements 216
11
HR is a PhD student in the GenomMed and Molecular Medicine (DPMM) Doctoral Programs 217
of the University of Eastern Finland, Kuopio, Finland. This study is part of the research 218
activities of the Finnish FTD Research Network (FinFTD).
219
Conflict of Interest 220
Pentti J. Tienari holds a patent on C9orf72 in diagnostics and treatment of ALS/FTLD. The 221
other authors have no conflicts of interest to disclose.
222
Funding 223
This work was supported by the Academy of Finland, under grant numbers 315459 (AH), 224
315460 (AMR), and 307866 (MH); the Strategic Neuroscience Funding of the University of 225
Eastern Finland; Finnish Brain Foundation (ES), Sigrid Juselius Foundation (ES, MH, PJT), 226
Finnish Cultural Foundation (KK) Instrumentarium Science Foundation (ES), Orion Research 227
Foundation (ES). This work was supported by a grant (European Alzheimer DNA BioBank, 228
EADB) from the EU Joint Programme – Neurodegenerative Disease Research (JPND). Inserm 229
UMR1167 is also funded by Inserm, Institut Pasteur de Lille, the Lille Métropole Communauté 230
Urbaine, the French government’s LABEX DISTALZ program (development of innovative 231
strategies for a transdisciplinary approach to Alzheimer’s disease). This publication is part of a 232
project that has received funding from the European Union’s Horizon 2020 research and 233
innovation programme under the Marie Skłodowska-Curie grant agreement No 740264.
234
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Table 1. Haplotypes and individual SNPs significantly associating with C9orf72 repeat expansion (C9exp) in the iNPH cohort and ALS and FTLD in the FinnGen cohort
Haplotype/
SNP
iNPH cohort (C9orf72 repeat expansion) Motor neuron disease ALS (FinnGen) Frontotemporal lobar degeneration (FinnGen)
OR (95% CI) p MAF in
C9exp carriers
MAF in control
OR (95% CI) p MAF in
disease
MAF in control
OR (95% CI) p MAF in
disease
MAF in control haplo 1 10.04 (5.79-17.39) 2.01×10-16 0.5119 0.1038 1.68 (1.34-2.12) 8.40×10-06 0.1933 0.1239 1.39 (1.09-1.77) 7.74×10-3 0.1653 0.1248 haplo 2 11.33 (6.38-20.14) 1.28×10-16 0.5000 0.0943 1.81 (1.43-2.28) 7.05×10-07 0.1849 0.1110 1.49 (1.16-1.90) 1.8×10-3 0.1570 0.1115 haplo 2* 42.74 (18.35-99.53) 3.16×10-18 0.1905 0.0071 5.26 (3.72-7.42) 4.05×10-21 0.0777 0.0156 4.41 (3.06-6.36) 1.97×10-15 0.0641 0.0159 haplo 3 7.08 (4.04-12.41) 7.59×10-12 0.3452 0.0802 1.94 (1.51-2.49) 2.72×10-07 0.1534 0.0847 1.59 (1.22-2.08) 0.7×10-3 0.1281 0.0857 haplo 4 7.08 (4.04-12.41) 7.59×10-12 0.3452 0.0802 1.94 (1.51-2.5) 2.57×10-07 0.1534 0.0846 1.59 (1.22-2.08) 6.77×10-4 0.1281 0.0856 haplo 5 8.23 (4.64-14.63) 6.45×10-13 0.3690 0.0820 2.12 (1.66-2.71) 1.65×10-09 0.1660 0.0860 1.62 (1.24-2.11) 4.23×10-4 0.1302 0.0862 haplo 5* 46.76 (19.71-110.9) 2.67×10-18 0.1905 0.0065 5.12 (3.61-7.26) 5.40×10-20 0.0756 0.0155 4.57 (3.19-6.56) 1.43×10-16 0.0661 0.0158 haplo 6 8.23 (4.64-14.63) 6.45×10-13 0.3690 0.0820 2.12 (1.66-2.71) 1.64×10-09 0.1660 0.0860 1.62 (1.24-2.11) 4.23×10-4 0.1302 0.0862 haplo 6* 46.76 (19.71-110.9) 2.67×10-18 0.1905 0.0065 5.12 (3.61-7.26) 5.40×10-20 0.0756 0.0155 4.57 (3.19-6.56) 1.43×10-16 0.0661 0.0158 haplo 7 7.12 (4.07-12.47) 6.73×10-12 0.3452 0.0796 1.94 (1.51-2.5) 2.40×10-07 0.1534 0.0844 1.57 (1.2-2.05) 1.15×10-3 0.1260 0.0854 haplo 8 9.48 (5.12-17.56) 8.87×10-13 0.3333 0.0696 2.15 (1.66-2.79) 8.82×10-09 0.1429 0.0714 1.74 (1.32-2.31) 9.32×10-5 0.1178 0.0720 haplo 8* 46.76 (19.71-110.9) 2.67×10-18 0.1905 0.0065 5.12 (3.61-7.27) 5.27×10-20 0.0756 0.0155 4.42 (3.07-6.38) 1.71×10-15 0.0641 0.0158 haplo 9 9.87 (5.32-18.32) 3.94×10-13 0.3333 0.0666 2.23 (1.72-2.89) 1.85×10-09 0.1429 0.0690 1.81 (1.37-2.39) 3.33×10-5 0.1178 0.0697 haplo 10 7.68 (4.13-14.3) 1.24×10-10 0.2500 0.0495 2.39 (1.83-3.13) 2.24×10-10 0.1303 0.0589 1.86 (1.39-2.5) 3.17×10-5 0.1054 0.0597 haplo 10* 46.51 (19.09-113.3) 2.88×10-17 0.1786 0.0059 5.68 (3.98-8.1) 1.12×10-21 0.0735 0.0137 4.84 (3.33-7.02) 1.13×10-16 0.0620 0.0140 rs139185008 39.41 (17.17-90.49) 4.58×10-18 0.1905 0.007665 5.19 (3.69-7.3) 2.57×10-21 0.0798 0.0163 4.38 (3.05-6.28) 1.08×10-15 0.0661 0.0166 rs3849942 8.44 (4.99-14.29) 2.04×10-15 0.5595 0.1486 1.58 (1.28-1.95) 1.94×10-05 0.2437 0.1698 1.22 (0.97-1.53) 8.53×10-2 0.1983 0.1704 Notes: iNPH cohort Ncarriers/control = 41/801; Motor neuron disease ALS Ncases/control = 238/111,855; Frontotemporal lobar degeneration Ncases/control = 242/214,474; * denotes that variant chr9_27491944_T_C (rs139185008) was added to haplotype analysis; chromosomal positions of SNPs constituting haplotypes are listed in Supplementary Table 1
Abbreviations: CI, confidence interval; FTLD, frontotemporal lobar degeneration; haplo, haplotype; iNPH, idiopathic normal pressure hydrocephalus; MAF, minor allele frequency; N, number of subjects; OR, odds ratio; SNP, single-nucleotide polymorphism
17
18
Figure 1: SNPs associating with the C9orf72 repeat expansion in iNPH cohort locate near the MOB3B and C9orf72 genes. Manhattan plot of genome-wide association (GWA) of SNPs associated with the C9orf72 expansion in a Finnish iNPH cohort. Chromosome numbers are indicated below the x-axis (A). Regional association plot of chromosome 9 locus, which contained significant association from the GWA study. SNPs of the previously described Finnish risk haplotype [12] and rs139185008 (arrow) are indicated above the plot. Significantly associated SNPs are indicated in bold. Linkage disequilibrium is indicated as color-coded r2 values. Recombination rates are depicted by continuous line. The reference variant rs3849942 is shown as a diamond. (B). Gray lines indicate significance level (p < 5×10-8). Abbreviations: iNPH, idiopathic normal pressure hydrocephalus; SNP, single nucleotide polymorphism
19
20
Figure 2: Geographical distribution of rs139185008 minor allele frequencies in different regions in Finland. Minor allele frequencies (MAF) of rs139185008 in Finland.
Pearson’s Chi-square test revealed statistically significant deviation (p < 2.2×10-16, X2 = 282.43, df = 18) of the geographical distribution of the minor allele counts of rs139185008. Genotyped population sizes are given for each region. Mean MAF for all regions is indicated as black vertical line (A). MAF of rs139185008 within Finnish regions showed geographical clustering of high (dark magenta) and low (white) frequencies (B).
Supplementary file
Supplementary Table 1. List of SNPs included in haplotypes
Supplementary Figure 1. Manhattan plot of genome-wide association (GWA) of SNPs associated with ALS in the UK Biobank
Supplementary Information. List of FinnGen authors and their affiliation
21 Supplementary Table 1. List of SNPs included in haplotypes
Haplotypes listed in Table 1; SNPs which are included in haplotypes are marked with alternative allele (gray); "*" in haplotype name denotes that SNP rs139185008 was added to haplotype analysis
position SNP reference alternative haplo 1 haplo 2 haplo 2* haplo 3 haplo 4 haplo 5 haplo 5* haplo 6 haplo 6* haplo 7 haplo 8 haplo 8* haplo 9 haplo 10 haplo 10*
27451484 rs4879507 C T C C
27453329 rs10812599 C T C C
27455825 rs10967945 C T C C
27456930 rs7021930 G A G G
27461738 rs76444167 A G A A
27463312 rs62542379 G A G G
27467672 rs10757663 G A A A A A A A A
27468264 rs61349511 A G A A A A A A A
27474216 rs10967952 T C T T T T T T T
27477876 rs1444533 C T T T T T T T T
27478054 rs1822723 C T C C C C C C C
27478711 rs4879514 T C C C C C C C C C C
27482237 rs4879515 C T T T T T T T T T T T T
27484498 rs10812602 A G A A A A A A A A A A A
27488094 rs17779457 T G G G G G G G G G G G
27489253 rs868856 A G A A A A A A A A A A A A A A A
27490969 rs7046653 A G A A A A A A A A A A A A A A A
27491944 rs139185008 T C C C C C C
27495475 G A G G G G G G G
27502988 rs1977661 C A C C C C C C C C C C C C C C C
27508689 rs2166128 T C C C C C C C C C C C
27510494 rs2477522 C T T T T T T T T T T T T
27513838 rs74439636 C T C C C C C C C C C C
27521398 rs75741240 C T C C C C C C C C C C
27527739 rs117257581 T C T T T T T T T T T T
27529318 rs903603 G A G G G G G G G G G G G
27533986 rs10812610 C A C C C C C C C C C C C
27536399 rs2814707 C T T T T T T T T T T T T T T T T
27543283 rs3849942 T C T T T T T T T T T T T T T T T
27543384 rs3849943 C T C C C C C C C C C C
27546892 rs13691 G A G G G G G G G G G G
27549487 rs80067552 G T G G G G G G G G G G
27553878 rs12349820 T C T T T T T T T T T T T T T T T
22
27556782 rs10122902 G A G G G G G G G G G G G G G G G
27556833 rs62538126 C A C C C C C C C C C C
27557532 rs76925759 T C T T T T T T T T T T
27557921 rs10757665 T C T T T T T T T T T T T
27559735 rs1565948 G A G G G G G G G G G G G
27561051 rs774359 T C C C C C C C C C C C C C C C C
27572257 rs2282241 C A C C C C C C C C C C C C C C C
27575787 rs1948522 C T C C C C C C C C C C C
27579562 rs1982915 A G G G G G G G G G G G G
27580676 rs12350076 A C C C C C C C C C C C C
27582805 rs17769370 C T C C C C C C C C C C
27583130 rs7864502 G C C C C C C C C C
27583296 rs702230 T A A A A A A A A A
27583756 rs28522676 G T T T T T T T T T
27583819 rs4879585 A C A A A A A A A A A A
27584061 rs34555425 G C G
27585699 rs36062268 G A G G G G G G G G G
27586164 rs2453556 A G G G G G G G G G G G G
27586447 rs7848063 G A A A A A A A A A A A A
27587790 rs3910852 T C T T T T T T T T T T
27588733 rs702231 C A A A A A A A A A A A A
27589659 rs696826 A G G G G G G G G G G G G
27589698 rs112001877 T C T T T T T T T T
27591128 rs9792688 T A T T
27601619 rs75747155 G A G G
23
Supplementary Figure 1. Manhattan plot of genome-wide association (GWA) of SNPs associated with ALS in the UK Biobank
