Volume 3 Supplement 7

Genetic Analysis Workshop 16

Open Access

Detecting susceptibility genes for rheumatoid arthritis based on a novel sliding-window approach

BMC Proceedings20093(Suppl 7):S14

DOI: 10.1186/1753-6561-3-S7-S14

Published: 15 December 2009

Abstract

With the recent rapid improvements in high-throughout genotyping techniques, researchers are facing a very challenging task of large-scale genetic association analysis, especially at the whole-genome level, without an optimal solution. In this study, we propose a new approach for genetic association analysis based on a variable-sized sliding-window framework. This approach employs principal component analysis to find the optimal window size. Using the bisection algorithm in window size searching, the proposed method tackles the exhaustive computation problem. It is more efficient and effective than currently available approaches. We conduct the genome-wide association study in Genetic Analysis Workshop 16 (GAW16) Problem 1 data using the proposed method. Our method successfully identified several susceptibility genes that have been reported by other researchers and additional candidate genes for follow-up studies.

Background

With the availability of large-scale genotyping technologies, the cost of genome-wide analyses has been greatly reduced and a boom of large-scale genetic association studies is underway. A sliding-window approach, in which several neighboring single-nucleotide polymorphisms (SNPs) together included in a "window frame", is a popular strategy of multiple allelic association analysis. During the test the window slides across the genome region under study in a stepwise fashion [13]. Variable sized sliding-window approaches with variable window sizes decided by the underlying linkage disequilibrium (LD) pattern perform more efficiently in large-scale data analysis. The problem for variable sized sliding-window approaches is how to search the optimal window size with being not only computationally practical but also statistically sufficient to gain higher detection power for both common and rare risk factors.

In this report, based on the variable sized sliding-window frame, we adapt the optimal window size to the local LD pattern by employing principal components (PC) approach. The PC approach is known as a linear projection method that defines a lower-dimensional space and captures the maximum information of the initial data [4]. Each optimal window size is defined by the first few PCs (i.e., 3 or 5) that could explain a main fraction of the total amount (i.e., 90% or 95%) of information in the data.

Data

In our study, we used the Genetic Analysis Workshop (GAW) 16 Problem 1 data, which is the initial batch of the whole-genome association data for the North American Rheumatoid Arthritis Consortium (NARAC). Data were available for 868 cases and 1194 controls. There are 22 chromosomes with 545,080 SNP-genotype fields from the Illumina 550k chip. To avoid the missing value problem, any subject who had missing values in that window was excluded from the current window. Thus, some subjects may not be in the current window but will still be included in the study in other windows. In this way, we retained the most information we could.

Methods

Optimal window size defined by PC analysis

We consider a study with total M individuals in a data set and with genotype information denoted by vectors G i = (gi1, gi2,, g iN ) T (i = 1,2,, M) at N SNP loci for the ith individual. We code the genotype g ij as 0, 1, or 2 for the number of minor (less frequent) alleles at SNP j, j = 1,2, N of individual i. Let y i denote the trait value of individual i.

In the sliding-window frame, a window denoted as https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif is a set of neighboring SNPs {b, b + 1, b + 2, , b + l - 1}. A variable sized sliding window which begins with SNP b, denoted as Ω b , is a collection of windows https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif with l ranging from s to Γ b , where s and Γ b are the smallest and largest window sizes.

In this study, we apply PC method to define the optimal window size. The basic idea is that we attempt to find the largest window size in which c0 proportion of the total information can be explained by the first k PCs and c0 and k are predefined criteria. We define this largest window size as the optimal window size. Start with a window https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif with l = s= k + 1, so that at least the window length is longer than the number of the important PCs.

Let https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq2_HTML.gif denote the sample variance-covariance matrix of genotypic numerical codes in window https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif and https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq3_HTML.gif denote the jth largest eigenvalue of https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq2_HTML.gif Thus, in window https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif , the total variance in the original dataset explained by the jth PC is https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq4_HTML.gif . Let https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq5_HTML.gif as the proportion of the total variability explained by the first k PCs. Our main idea of choosing the optimal window size of each sliding window is to find the largest window size in which c0 proportion of the total variability can be explained by the first k PCs among a set of windows Ω b .

Bisection method for searching the optimal window size and computational consideration

Using the exhaustive searching method may be computational demanding for determining the optimal window size. We propose to use bisection method. Let s and Γ denote the predefined smallest and largest window sizes among a set of windows Ω b , where b is the starting SNP of the set of windows.

By adapting bisection method, the searching procedure for the optimal window size in Ω b includes following steps:

Step 1: Let l be the middle point of s and Γ, that is, l = [(s = Γ)2/], where [a] is the largest integer that is less than or equal to a.

Step 2: Conduct PC analysis within the window https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif , where a window begins at SNP b and has a size l.

Step 3: Calculate C (the proportion of the total variability explained by the first k PCs) for the window https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif . If C > c0, we let s = l, that is, we update the smallest window size s. Otherwise, we let Γ = l, that is, we update the largest window size Γ.

Step 4: Repeat Step 1 to Step 3 until Γ - s ≤ 1.

In the window https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq6_HTML.gif , if the proportion of the total variability explained by the first k PCs is greater than c0, the optimal window size will be Γ; otherwise, the optimal window size will be s.

Until now, we have not mentioned how to choose the starting SNP b. Of course for the first window, b = 1. To choose b for other windows, the following three methods are typically used. For the ith (i > 1) window, choose 1) b = i; 2) b = n i , where n i is the middle SNP of the (i-1)th window; 3) b = m i +1, where m i is the last SNP of the (i-1)th window. In this article, we use the first method to choose the starting SNP b.

By using bisection method, our proposed variable length sliding-window method is computationally efficient. Consider a set of windows Ω b with the smallest window size s, largest window size Γ, and starting SNP b. The computational complexity to find the optimal window size in Ω b using the bisection algorithm is Γ3log2(Γ - s). If we have N SNPs in total, the computational complexity to find all the optimal window sizes is NΓ3log2(Γ - s). In this article, we use Γ = 35 and s = 4. Suppose N = 500,000 in a genome-wide association study. Then, NΓ3log2(Γ - s) <N2. As pointed out by one of the reviewers, HAPLOVIEW program may be used to find beginning and end of a window. Using HAPLOVIEW, N2 of pair-wise r2 need to be calculated. To calculate r2, we need to estimate haplotype frequencies. Theoretically, our proposed method should be computationally more efficient than HAPLOVIEW. In fact, we have done a preliminary simulation study. The results show that the computation time of our proposed method is about a hundred times faster than HAPLOVIEW.

Score test

After we find the optimal window size for each sliding window, we use the score test statistic based on a logistic model [5] to test for association within each sliding window. Consider https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq1_HTML.gif , a window beginning at SNP b with an optimal window size l. Take b = 1 as an example for windows that start at the first SNP. Let https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq7_HTML.gif denote its first k PCs of the ith individual, where i = 1,2,, M. Suppose that the k PCs follow a logistic model, then, the score test statistic is given by T2 = U'V-1U, where https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq8_HTML.gif , https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq9_HTML.gif , https://static-content.springer.com/image/art%3A10.1186%2F1753-6561-3-S7-S14/MediaObjects/12919_2009_Article_2761_IEq10_HTML.gif , and M is the sample size. The statistic T2 asymptotically follows a χ2 distribution with k degrees of freedom. We select significant windows after adjusting for multiple testing using a Bonferroni correction.

Result

We applied the proposed approach to GAW16 Problem 1. In our application, we set s = 4; Γ = 35; c0 = 90% and k = 3. Originally the dataset contained genotypes at 545,080 SNPs on chromosomes 1 to 22. In our analysis we ended up with 531,501 windows. The size of the windows varied from 4 to 29 SNPs, with the median window size of 7 (see Table 1 for the distribution of the window sizes). After Bonferroni correction, we found 1,155 significant windows. Due to the strong LD among SNPs, many of the significant windows overlapped with the nearby windows. In order to report the result thoroughly, we combined the significant windows with all overlapped windows as one larger window. Thus, we end up with 76 significant larger windows. Due to the limited pages, in Table 2 we only report the top 30 windows after the combination. The order of the windows is according to their most significant sub-windows (the original window before combinations). Our result matches most of the genes reported in recent studies [611] and also identify more rheumatoid arthritis (RA) susceptibility genes for follow-up studies.
Table 1

The distribution of the window sizes based on 531,501 windows

Window size

Percentage of total windows

4

18.32

5

11.92

6

11.69

7

11.57

8

8.94

9

7.34

10

6.34

11

4.67

12

3.73

13-29

15.48

Table 2

Genetic and physical map locations of window region identified using PC-sliding-window analysis based on the Bonferroni correction

Window ID

Chra

Physical location

Genesb

CRASGc

1

6

30014670, 33187144

TNF, HLA-A HLA-B, HLA-C

TNF, HLA-A HLA-B, HLA-C

2

1

792429, 1101089

AGRN, C1orf159, ISG15, SAMD11

 

3

2

172768404, 172807000

DLX1, DLX2

STAT4, ITGAV

4

12

46666298, 46718200

COL2A1, LOC728181 LOC728114

 

5

13

113656958, 113861908

FAM70B, RASA3

 

6

7

154133201, 154241160

PAXIP1, LOC202781

 

7

17

68283979, 68361160

SLC39A11

 

8

2

98261543, 98370780

VWA3B

IL1B

9

13

49336428, 49340230

KPNA3

 

10

1

2243956, 3359357

ARHGEF16, PRDM16

 

11

17

66647226, 66750860

Intergenic 17q24

 

12

16

67482002, 67660490

TMC07, FLJ12331

 

13

18

75300466, 75314140

NFATC1

 

14

13

74883232, 74941310

TBC1D4

 

15

9

123211883, 123248000

CRB2, MIRN601, DENND1A

TRAF1/C5

16

20

57796484, 57832810

  

17

1

15181683, 151846600

ADAM15, EFNA4

 

18

20

35438689, 35501280

SRC, RPL7AL4

 

19

22

28164734, 28237820

RFPL1S, RFPL1, NEFH

 

20

11

64593946, 64661480

SAC3D1, NAALADL1 CDCA5, ZFPL1, ZHIT2

 

21

11

45207308, 45314100

SYT13, FLJ41423

 

22

8

20327035, 20435200

  

23

5

137614229, 137825000

GFRA3, CDC25C, FAM53C, JMJD1B

 

24

7

129525353, 129580000

CPA2, CPA4

 

25

3

134954925, 134966000

TF, SRPRB

 

26

9

104811123, 104815000

ABCA1

 

27

12

6924169, 6932652

ATN1, CL2orf57, PTPN6

 

28

10

10531181, 10542841

SH3PXD2A

 

29

11

3335218, 3524620

ZNF195, OR7Z12p

 

30

19

19106771, 19154190

TMEM16A1, MEF2B

 

aChr, chromosome

bWe found the significant genes using the NCBI dbSNP database http://www.ncbi.nlm.nih.gov/sites/entrez?db=snp.

cThe confirmed RA susceptibility genes (CRASG) are shown in the last column if they are within or near our significant region.

Discussion

As the most exhaustive searching engine in genome-wide association studies, sliding-window approaches are receiving more and more attention recently. Based on the variable sized sliding-window frame, we adapt the optimal window size to the local LD pattern by employing the PC approach. We applied this novel sliding-window approach to the GAW16 RA data and successfully validated nine genes that have been reported by recent studies and also identified new candidate genes for follow-up studies.

Our approach has several advantages. It provides a stable method to choose the window size with the maximum information extraction and it automatically balances degrees of freedom and number of tests, which results in higher power to detect association. It is flexible enough to conduct different association tests within the windows. The method is computational efficient when applied to large-scale data compared with other variable sized sliding-window methods. It requires only genotype data so there is no need to go through any computationally intensive phasing program to account for uncertain haplotype phases.

Further efforts are needed to improve the proposed method, such as determining the optimal c0 (the proportion of the total variability explained by the top k PCs) and the initial window lengths in the bisection method.

Conclusion

In this study, we applied our novel genome-wide PC sliding-window approach to detect the association between SNP windows and disease status using GAW16 Problem 1 RA dataset. We validated nine genes which have been identified to be responsible for RA in the literature and discovered more genes and non-gene regions for follow-up studies.

List of abbreviations used

GAW: 

Genetic Analysis Workshop

LD: 

Linkage disequilibrium

NARAC: 

North American Rheumatoid Arthritis Consortium

PC: 

Principal components

RA: 

Rheumatoid arthritis

SNP: 

Single-nucleotide polymorphism

Declarations

Acknowledgements

The Genetic Analysis Workshops are supported by NIH grant R01 GM031575 from the National Institute of General Medical Sciences.

This work was supported by NIH grants R01 GM069940 and the Overseas-Returned Scholars Foundation of Department of Education of Heilongjiang Province (1152HZ01).

This article has been published as part of BMC Proceedings Volume 3 Supplement 7, 2009: Genetic Analysis Workshop 16. The full contents of the supplement are available online at http://www.biomedcentral.com/1753-6561/3?issue=S7.

Authors’ Affiliations

(1)
Department of Mathematical Sciences, Michigan Technological University
(2)
Mathematical Sciences, Heilongjiang University

References

  1. Yang HC, Lin CY, Fann CSJ: A sliding-window weighted linkage disequilibrium test. Genet Epidemiol. 2006, 30: 531-545. 10.1002/gepi.20165.View ArticlePubMedGoogle Scholar
  2. Li Y, Sung W, Liu JJ: Association mapping via regularized regression analysis of single-nucleotide-polymorphism haplotypes in variable-sized sliding windows. Am J Hum Genet. 2007, 80: 705-715. 10.1086/513205.PubMed CentralView ArticlePubMedGoogle Scholar
  3. Huang BE, Amos CI, Lin DY: Detecting haplotype effects in genomewide association studies. Genet Epidemiol. 2007, 31: 803-812. 10.1002/gepi.20242.View ArticlePubMedGoogle Scholar
  4. Sha Q, Dong J, Jiang R, Zhang S: Test of association between quantitative traits and haplotypes in a reduced-dimensional space. Ann Hum Genet. 2005, 69: 715-732. 10.1111/j.1529-8817.2005.00216.x.View ArticlePubMedGoogle Scholar
  5. Chapman JM, Cooper JD, Todd JA, Clayton DG: Detecting disease associations due to linkage disequilibrium using haplotype tags: a class of tests and the determinants of statistical power. Hum Hered. 2003, 56: 18-31. 10.1159/000073729.View ArticlePubMedGoogle Scholar
  6. Begovich AB, Carlton VE, Honigberg LA, Schrodi SJ, Chokkalingam AP, Alexander HC, Ardlie KG, Huang Q, Smith AM, Spoerke JM, Conn MT, Chang M, Chang SY, Saiki RK, Catanese JJ, Leong DU, Garcia VE, McAllister LB, Jeffery DA, Lee AT, Batliwalla F, Remmers E, Criswell LA, Seldin MF, Kastner DL, Amos CI, Sninsky JJ, Gregersen PK: A missense single-nucleotide polymorphism in a gene encoding a protein tyrosine phosphatase (PTPN22) is associated with rheumatoid arthritis. Am J Hum Genet. 2004, 75: 330-337. 10.1086/422827.PubMed CentralView ArticlePubMedGoogle Scholar
  7. Thomson W, Barton A, Ke X, Eyre S, Hinks A, Bowes J, Donn R, Symmons D, Hider S, Bruce IN, Wellcome Trust Case Control Consortium, Wilson AG, Marinou I, Morgan A, Emery P, YEAR Consortium, Carter A, Steer S, Hocking L, Reid DM, Wordsworth P, Harrison P, Strachan D, Worthington J: Rheumatoid arthritis association at 6q23. Nat Genet. 2007, 39: 1431-1433. 10.1038/ng.2007.32.PubMed CentralView ArticlePubMedGoogle Scholar
  8. Remmers EF, Plenge RM, Lee AT, Graham RR, Hom G, Behrens TW, de Bakker PI, Le JM, Lee HS, Batliwalla F, Li W, Masters SL, Booty MG, Carulli JP, Padyukov L, Alfredsson L, Klareskog L, Chen WV, Amos CI, Criswell LA, Seldin MF, Kastner DL, Gregersen PK: STAT4 and the risk of rheumatoid arthritis and systemic lupus erythematosus. N Engl J Med. 2007, 357: 977-986. 10.1056/NEJMoa073003.PubMed CentralView ArticlePubMedGoogle Scholar
  9. Plenge RM, Seielstad M, Padyukov L, Lee AT, Remmers EF, Ding B, Liew A, Khalili H, Chandrasekaran A, Davies LR, Li W, Tan AK, Bonnard C, Ong RT, Thalamuthu A, Pettersson S, Liu C, Tian C, Chen WV, Carulli JP, Beckman EM, Altshuler D, Alfredsson L, Criswell LA, Amos CI, Seldin MF, Kastner DL, Klareskog L, Gregersen PK: TRAF1-C5 as a risk locus for rheumatoid arthritis-a genomewide study. N Engl J Med. 2007, 357: 1199-1209. 10.1056/NEJMoa073491.PubMed CentralView ArticlePubMedGoogle Scholar
  10. Kurreeman FA, Padyukov L, Marques RB, Schrodi SJ, Seddighzadeh M, Stoeken-Rijsbergen G, Helm-van Mil van der AH, Allaart CF, Verduyn W, Houwing-Duistermaat J, Alfredsson L, Begovich AB, Klareskog L, Huizinga TW, Toes RE: A candidate gene approach identifies the TRAF1/C5 region as a risk factor for rheumatoid arthritis. PLoS Med. 2007, 4: e278-10.1371/journal.pmed.0040278.PubMed CentralView ArticlePubMedGoogle Scholar
  11. Jacq L, Garnier S, Dieudé P, Michou L, Pierlot C, Migliorini P, Balsa A, Westhovens R, Barrera P, Alves H, Vaz C, Fernandes M, Pascual-Salcedo D, Bombardieri S, Dequeker J, Radstake TR, Van Riel P, Putte van de L, Lopes-Vaz A, Glikmans E, Barbet S, Lasbleiz S, Lemaire I, Quillet P, Hilliquin P, Teixeira VH, Petit-Teixeira E, Mbarek H, Prum B, Bardin T, Cornélis F, European Consortium on Rheumatoid Arthritis Families: The ITGAV rs3738919-C allele is associated with rheumatoid arthritis in the European Caucasian population: a family-based study. Arthritis Res Ther. 2007, 9: R63-10.1186/ar2221.PubMed CentralView ArticlePubMedGoogle Scholar

Copyright

© Sha et al; licensee BioMed Central Ltd. 2009

This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Advertisement