ONLINE SUPPLEMENT ASSOCIATION OF KCNJ1 VARIATION WITH CHANGE IN FASTING GLUCOSE AND NEW ONSET DIABETES DURING HCTZ TREATMENT Jason H Karnes, PharmD 1, Caitrin W McDonough, PhD 1, Yan Gong, PhD 1, Teresa T Vo 1, Taimour Y Langaee, PhD 1, Arlene B Chapman, MD 2, John G Gums, PharmD 1,3, Amber L Beitelshees, PharmD, MPH 4, Kent R Bailey, PhD 5, Jorge L Del-Aguila, MD 6, Eric A Boerwinkle, PhD 6, Carl J Pepine, MD 3, Stephen T Turner, MD 2, Julie A Johnson, PharmD 1,3, and Rhonda M Cooper-DeHoff, PharmD, MS 1,3 1. University of Florida College of Pharmacy, Gainesville, FL 2. Emory University School of Medicine, Atlanta, GA 3. University of Florida College of Medicine, Gainesville, FL 4. University of Maryland School of Medicine, Baltimore, MD 5. Mayo Clinic College of Medicine, Rochester, MN 6. University of Texas at Houston Center for Human Genetics, Houston, TX Corresponding Author: Rhonda M Cooper-DeHoff, PharmD, MS Department of Pharmacotherapy and Translational Research Center for Pharmacogenomics College of Pharmacy, University of Florida 1600 SW Archer Road PO Box 100486 Gainesville, FL 32610-0486 Phone: 352-273-6184 FAX: 352-273-6121 E-mail: dehoff@cop.ufl.edu
Table S1. Gene location, minor allele frequencies, and Hardy Weinberg equilibrium p values for KCNJ1 SNPs in PEAR SNP Location Black Non-black MAF HWE 1 MAF HWE 1 rs675388 3 UTR 0.16 0.20 0.16 0.68 rs1231254 3 UTR 0.82 0.49 0.16 0.69 rs2238009 Intron 0.41 0.69 0.12 0.60 rs6590353 Intron 0.02 0.79 0 - rs1148058 Intron 0.09 0.35 0.17 0.22 rs1148059 Intron 0.18 0.27 0.17 0.25 rs3758766 Intron 0.09 0.35 0.17 0.25 rs17137967 Intron 0.05 0.11 0 - rs581472 Intron 0.37 0.02 0.35 0.55 rs2301152 Intron 0-0.001 0.98 rs7120491 Intron 0.12 0.20 0.001 0.98 rs583352 Intron 0.40 0.01 0.34 0.60 rs7933427 Intron 0.05 0.33 0.04 0.01 rs675482 Intron 0.42 0.84 0.27 0.77 rs658903 Intron 0.05 0.35 0.15 0.06 rs2846680 Intron 0.19 0.63 0.22 0.31 rs12795437 Intron 0.01 0.86 0.07 0.89 rs2855800 Intron 0.29 0.94 0.26 0.18 rs11600347 Intron 0.01 0.84 0.07 0.89 rs2855794 Intron 0.45 0.72 0.32 0.09 rs2186832 Intron 0.19 0.35 0.19 0.66 rs3016774 Intron 0.17 0.55 0.19 0.59 rs59172778 Exon 0-0.01 0.81 rs2855790 5 UTR 0.18 0.93 0.19 0.62 rs7116606 5 Upstream 0.37 0.09 0.29 0.89 HWE indicates Hardy Weinberg equilibrium p value; MAF, minor allele frequency; SNP, single nucleotide polymorphism; UTR, un-translated region. 1 Hardy Weinberg Equilibrium p value calculated according to Fisher s Exact Test.
Table S2. Association of linear regression model covariates and change in fasting glucose in PEAR Covariate Parameter Estimate 1 p value 1 (standard error) Baseline fasting glucose (mg/dl) -0.39 (0.04) <0.0001 Age (years) 0.09 (0.05) 0.09 Gender (male) -0.28 (0.93) 0.76 Waist circumference (cm) 0.18 (0.03) <0.0001 Potassium supplementation 0.37 (1.48) 0.80 Randomized drug arm (HCTZ) -0.44 (1.01) 0.67 Systolic blood pressure (mmhg) 2 0.03 (0.05) 0.62 Diastolic blood pressure (mmhg) 2-0.06 (0.09) 0.52 Time of HCTZ treatment (days) -0.07 (0.03) 0.05 Race/ethnicity (Black) -0.62 (5.97) 0.92 Principal component 1 26.73 (80.37) 0.74 Principal component 2 3.02 (11.98) 0.80 Principal component 3-7.74 (13.00) 0.55 HCTZ indicates hydrochlorothiazide; mg/dl, milligrams per deciliter. 1 Generated using linear regression of change in fasting glucose for all PEAR patients. 2 Represents average of home blood pressure values.
Table S3. Minor allele frequencies, and Hardy Weinberg equilibrium p values for KCNJ1 SNPs in INVEST White Hispanic Black SNP MAF HWE 1 MAF HWE a MAF HWE 1 rs675388 0.15 0.19 0.14 0.85 0.18 0.52 rs1231254 0.15 0.19 0.15 0.59 0.20 1 rs2238009 0.15 0.69 0.24 0.54 0.44 0.71 rs6590353 0-0.01 1 0.02 0.06 rs1148058 0.17 1 0.16 0.33 0.08 1 rs1148059 0.17 0.86 0.18 0.35 0.14 0.13 rs3758766 0.17 1 0.16 0.33 0.08 1 rs17137967 0-0.01 1 0.05 1 rs581472 0.37 0.46 0.45 0.65 0.37 1 rs2301152 0-0 - 0 - rs7120491 0-0.03 0.45 0.12 0.37 rs583352 0.37 0.46 0.45 0.78 0.38 1 rs7933427 0.05 1 0.07 0.50 0.06 1 rs675482 0.31 0.41 0.31 0.40 0.42 0.58 rs658903 0.16 0.06 0.12 1 0.06 1 rs2846680 0.23 0.89 0.26 0.13 0.24 0.46 rs12795437 0.07 0.71 0.10 0.81 0.01 1 rs2855800 0.27 0.61 0.34 0.06 0.32 1 rs11600347 0.07 0.71 0.10 0.81 0.02 1 rs2855794 0.34 0.58 0.45 1 0.46 0.71 rs2186832 0.19 0.15 0.22 0.69 0.24 0.46 rs3016774 0.19 0.15 0.21 1 0.21 1 rs59172778 0.01 1 0.01 1 0 - rs2855790 0.19 0.11 0.21 0.79 0.21 1 rs7116606 0.29 0.55 0.32 0.76 0.32 0.68 HWE indicates Hardy Weinberg Equilibrium p value; MAF, minor allele frequency; SNP, single nucleotide polymorphism. 1 Hardy Weinberg equilibrium p value determined by Fisher s Exact test in controls.
Table S4. Association of logistic regression model covariates and new onset diabetes in INVEST Characteristic Odds ratio (95% Confidence Interval) p value Baseline Age (years) 1.00 (0.99-1.02) 0.54 Gender (female) 0.92 (0.71-1.20) 0.54 BMI (kg/m 2 ) 1.06 (1.03-1.08) <0.0001 Hypercholesterolemia 1 1.06 (0.83-1.36) 0.63 History of smoking 1.08 (0.84-1.40) 0.55 Principal component 1 45.04 (0.12- >999) 0.21 Principal component 2 <0.001 (<0.001-0.10) 0.006 Principal component 3 0.08 (<0.001-12.63) 0.33 During INVEST Systolic blood pressure (mmhg) 2 1.01 (1.00-1.02) 0.02 Atenolol treatment 1.08 (0.85-1.37) 0.51 Trandolapril treatment 0.68 (0.53-0.88) 0.003 Potassium supplementation 1.93 (1.34-2.77) 0.0004 BMI indicates body mass index; INVEST, INternational VErapamil SR and Trandolapril Study. 1 History of or currently taking lipid-lowering medications. 2 Average of clinic blood pressure measurements during study.
Figure S1: Odds ratios per copy of allele and 95% confidence intervals for KCNJ1 SNPs nominally associated (p<0.05) with new onset diabetes during hydrochlorothiazide treatment in INVEST patients by race/ethnicity. All odds ratios are adjusted for age, gender, body mass index, average on treatment systolic blood pressure, hypercholesterolemia, history of smoking, potassium supplementation, principal components one, two, and three, and trandolapril or atenolol treatment. NOD indicates new onset diabetes; SNP, single nucleotide polymorphism.
Figure S2: Haploview-generated linkage disequilibrium (LD) plot of KCNJ1 SNPs in PEAR whites. Regions of higher LD are shaded darker according to higher r 2 values. The number within each box indicates the r 2 value. Monomorphic SNPs are not included.
Figure S3: Haploview-generated linkage disequilibrium (LD) plot of KCNJ1 SNPs in PEAR blacks. Regions of higher LD are shaded darker according to higher r 2 values. The number within each box indicates the r 2 value. Monomorphic SNPs are not included.
Figure S4: Haploview-generated linkage disequilibrium (LD) plot of KCNJ1 SNPs in INVEST whites. Regions of higher LD are shaded darker according to higher r 2 values. The number within each box indicates the r 2 value. Monomorphic SNPs are not included.
Figure S5: Haploview-generated linkage disequilibrium (LD) plot of KCNJ1 SNPs in INVEST Hispanics. Regions of higher LD are shaded darker according to higher r 2 values. The number within each box indicates the r 2 value. Monomorphic SNPs are not included.
Figure S6: Haploview-generated linkage disequilibrium (LD) plot of KCNJ1 SNPs in INVEST Blacks. Regions of higher LD are shaded darker according to higher r 2 values. The number within each box indicates the r 2 value. Monomorphic SNPs are not included.