Title: Association of CDX1 Binding Site of Periostin Gene with Bone Mineral Density And

Title: Association of CDX1 Binding Site of Periostin Gene with Bone Mineral Density and Vertebral Fracture Risk

Authors: Su-Mei Xiao1,2, Yi Gao1,Ching-Lung Cheung1, Cora H Bow1, Kam-Shing Lau1, Pak C Sham3,4, Kathryn CB Tan1,2,Annie WC Kung1,2

Institutions: Department of Medicine1, and Research Centre of Heart, Brain, Hormone & Healthy Aging2, and Department of Psychiatry3, and Centre for Reproduction, Development and Growth4, Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital, Hong Kong, China

Running title: Periostin and osteoporosis

Corresponding author:

Su-Mei Xiao, PhD

Department of Medicine,

Faculty of MedicineBuilding,

The University of Hong Kong,

Hong Kong,

China

Phone: (852)–2819 9749

Fax: (852) – 28162095

Email:

Table S1. Basic characteristics of HKSC cohort with extreme BMD (n=1,572)

Total subjects / Lumbar spine Group / Femoral neck Group
High BMD Group / Low BMD Group / P / High BMD Group / Low BMD Group / P / High BMD Group / Low BMD Group / P
Subjects number / 663 / 909 / 425 / 619 / 412 / 520
Age (years) / 47.7 (15.46) / 50.5 (16.02) / <0.05 / 49.6 (15.31) / 51.7 (14.68) / <0.05 / 46.6 (15.71) / 48.5 (17.07) / 0.08
Height (m) / 1.61 (0.08) / 1.55 (0.08) / <0.01 / 1.61 (0.08) / 1.55 (0.08) / <0.01 / 1.60 (0.08) / 1.54 (0.08) / <0.01
Weight (kg) / 63.59 (10.91) / 50.73 (8.66) / <0.01 / 63.46 (10.79) / 51.47 (8.96) / <0.01 / 64.77 (11.15) / 48.85 (7.28) / <0.01
BMD (g/cm2)
Lumbar spine / 1.09 (0.13) / 0.74 (0.13) / <0.01 / 1.15 (0.10) / 0.71 (0.09) / <0.01 / 1.08 (0.14) / 0.76 (0.14) / <0.01
Femoral neck / 0.86 (0.12) / 0.58 (0.09) / <0.01 / 0.83 (0.13) / 0.60 (0.09) / <0.01 / 0.91 (0.10) / 0.54 (0.08) / <0.01
BMD z-score
Lumbar spine / 1.16 (0.84) / -1.43 (0.68) / <0.01 / 1.62 (0.62) / -1.76 (0.42) / <0.01 / 1.03 (0.98) / -1.31 (0.75) / <0.01
Femoral neck / 1.10 (0.82) / -1.25 (0.66) / <0.01 / 0.92 (0.91) / -1.10 (0.71) / <0.01 / 1.58 (0.56) / -1.68 (0.35) / <0.01

Data are expressed as mean (SD). The T-test was conducted for phenotype comparison between high BMD group and low BMD group.

Table S2. Association results of common haplotype with BMD variation in all of the 1,572 HKSC subjects with extreme BMD

Global test (P) / Haplotype / Frequency / OR / P
0.038 / CGCTGAAC / 0.07 / 1.31 / >0.1
CGTTGAGG / 0.06 / 1.35 / >0.1
CGCTGAGC / 0.28 / 0.83 / 0.049
CGTTGAAG / 0.13 / 1.34 / 0.025
CACTGAAC / 0.33 / 0.85 / >0.1
TGCCGTAG / 0.07 / 1.13 / >0.1
CGCCAAAG / 0.03 / 0.75 / >0.1

The results were adjusted for age, height, weight and gender. The direction of haplotype follows the sequence of SNPs listed in the table 2.

Table S3. Association results of POSTNand SOSTgenes with BMD variation in the GWAS of Hong Kong Southern Chinese with extreme BMD (n=800 females)

Either LS or FN / LS / FN
SNP / Chr / Position / MAF / A1 / A2 / P / OR / 95%CI / P / OR / 95%CI / P / OR / 95%CI
POSTN
rs9547947 / 13 / 37029993 / 0.029 / A / C / >0.1 / 0.92 / 0.43-1.95 / >0.1 / 1.14 / 0.46-2.82 / >0.1 / 0.54 / 0.19-1.57
rs9547952 / 13 / 37036689 / 0.074 / T / C / 0.065 / 1.59 / 0.97-2.59 / >0.1 / 1.45 / 0.77-2.74 / >0.1 / 1.53 / 0.83-2.82
rs9315503 / 13 / 37037260 / 0.189 / G / A / 0.010 / 1.59 / 1.12-2.28 / 0.006 / 1.82 / 1.19-2.80 / >0.1 / 1.23 / 0.75-2.02
rs9603226 / 13 / 37041586 / 0.358 / A / G / >0.1 / 0.87 / 0.66-1.15 / >0.1 / 0.81 / 0.58-1.14 / >0.1 / 1.02 / 0.70-1.49
rs7323378 / 13 / 37051351 / 0.101 / C / T / >0.1 / 1.34 / 0.89-2.02 / >0.1 / 1.31 / 0.79-2.18 / >0.1 / 1.17 / 0.69-1.99
rs6563562 / 13 / 37056718 / 0.074 / C / T / 0.065 / 1.59 / 0.97-2.59 / >0.1 / 1.45 / 0.77-2.74 / >0.1 / 1.53 / 0.83-2.82
rs12871092 / 13 / 37057633 / 0.351 / G / A / 0.056 / 0.77 / 0.59-1.01 / >0.1 / 0.78 / 0.56-1.07 / >0.1 / 0.83 / 0.59-1.18
rs2025405 / 13 / 37068523 / 0.291 / A / G / 0.003 / 1.57 / 1.17-2.11 / 0.003 / 1.73 / 1.21-2.47 / >0.1 / 1.20 / 0.81-1.79
rs2985159 / 13 / 37074489 / 0.189 / G / T / 0.014 / 1.56 / 1.09-2.22 / 0.006 / 1.83 / 1.19-2.81 / >0.1 / 1.16 / 0.71-1.91
SOST
rs9899889 / 17 / 39178751 / 0.388 / G / T / >0.1 / 0.91 / 0.69-1.20 / >0.1 / 0.97 / 0.70-1.34 / >0.1 / 0.77 / 0.52-1.12
rs865429 / 17 / 39190741 / 0.246 / C / T / >0.1 / 1.25 / 0.93-1.68 / >0.1 / 1.29 / 0.91-1.84 / >0.1 / 1.38 / 0.92-2.10
rs1234612 / 17 / 39196328 / 0.139 / C / T / >0.1 / 1.32 / 0.92-1.91 / >0.1 / 1.20 / 0.78-1.84 / 0.041 / 1.72 / 1.02-2.89
rs2301682 / 17 / 39213552 / 0.126 / T / G / >0.1 / 1.31 / 0.89-1.94 / >0.1 / 1.13 / 0.71-1.81 / 0.054 / 1.74 / 0.99-3.06

Genomic position, B36; A1, minor/effectallele; A2, major allele; MAF, minor allele frequency;LS, lumbar spine; FN, femoral neck; The results were adjusted for age, height, weight.OR >1, theeffectallele is associated with the higher risk of low BMD.

FigureS1. The genomic location (B36) and pair-wise linkage disequilibrium (LD) coefficients (r2) of eight polymorphic SNPs in the POSTN gene. Vertical bars represent exons. Blocks connecting SNP pairs are shaded according to the LD strength between SNPs by using the r2, which are ranged from 0 (white) to 1.0 (dark black).

Figure S2. The measured raw BMD value (mean±SE) at lumbar spine (LS) and femoral neck (FN)for AAand GG genotype groups of SNP rs9547970 in the Hong Kong Osteoporosis Study prospective cohort (n = 2,509). The raw BMD value was 0.030 and 0.011 (g/cm2) less in minor allele GG carriers compared with AA carriers for LS and FN, respectively.