Online Appendix for the following October 28 JACC article
TITLE: Naturally Occurring Human Genetic Variation in the 3'-Untranslated Region of the Secretory
Protein Chromogranin A Is Associated With Autonomic Blood Pressure Regulation and Hypertension in a
Gender-Dependent Fashion
AUTHORS: Yuqing Chen, MD, PHD, Fangwen Rao, MM, Juan L. Rodriguez-Flores, MS, Manjula
Mahata, PHD, Maple M. Fung, MD, Mats Stridsberg, PHD, Sucheta M. Vaingankar, PHD, Gen Wen, MD,
Rany M. Salem, MPH, Madhusudan Das, PHD, Myles G. Cockburn, PHD, Nicholas J. Schork, PHD,
Michael G. Ziegler, MD, Bruce A. Hamilton, PHD, Sushil K. Mahata, PHD, Laurent Taupenot, PHD,
Daniel T. O’Connor, MD
APPENDIX
Online Figure I. Patterns of Linkage Disequilibrium at the Human CHGA Locus
Data are shown for 13 common (minor allele frequency >5%) biallelic polymorphisms spanning the
CHGA gene, discovered by systematic resequencing with amplicons encompassing each exon,
exon/intron border, 5'- and 3'-UTR, and proximal promoter (21). Pairwise results are plotted on a
pseudocolor scale for the linkage disequilibrium (LD) parameter D', with the GOLD (Graphical
Observation of Linkage Disequilibrium) algorithm, for subjects self-identified as white (European
ancestry, 2n = 102 chromosomes).
Online Figure II. CHGA 3'-UTR Common Variant C+87T and BP in Population
Panel A. CHGA 3'-UTR common variant C+87T: Sex-dependent effect on blood pressure (BP) status
(dichotomous trait) in population BP extremes. Results are shown separately for men and women. Alleles:
C = 73%, T = 27%; HWE: 2 = 0.286, p = 0.593. Panel B. CHGA 3'-UTR common variant C+87T: Sexdependent effect on BP as a quantitative trait in population BP extremes. Data were analyzed by ANOVA
(dependent variables: DBP or SBP, in mm Hg), in order to accommodate the covariate of sex, and to
estimate gene-by-sex interactions on the trait. Results are shown separately for men and women.
On-line Table I. CHGA Genotype and BP Study: Subject Characteristics. 1019 individuals of
European (white) ancestry, both male and female, were selected from the most extreme (upper and lower
5th percentiles) blood pressures in a primary care population of >53,000 individuals. Hypertension was
treated by antihypertensive medications in 48% of subjects with hypertension. Subjects were phenotyped
for not only blood pressure but also metabolic and renal traits. These individuals then gave rise to the
CHGA genetic case/control study.
Sex
2-Way ANOVA
Male
BP Status
Female
BP Status
Sex
BP-by-Sex
p Value
p Value
p Value
153.5 ± 1.0
<0.001
<0.001
0.006
55.1 ± 0.3
97.4 ± 0.3
<0.001
<0.001
0.213
73.3 ± 0.7
65.6 ± 0.6
73.7 ± 0.8
<0.001
0.002
0.01
25.7 ± 1.2
30.3 ± 1.1
24.0 ± 1.0
30.3 ± 1.1
<0.001
0.08
0.8
196.1 ± 2.7
218.8 ± 2.6
206.5 ± 2.3
228.8 ± 2.6
<0.001
<0.001
0.938
48.5 ± 1.4
47.0 ± 1.2
66.3 ± 1.1
60.6 ± 1.2
0.03
<0.001
0.085
1.10 ± 0.01
1.07 ± 0.01
0.87 ± 0.01
0.86 ± 0.01
0.771
<0.001
0.213
Normotensive
Hypertensive
Normotensive
Hypertensive
249
221
230
319
SBP, mm Hg
111.8 ± 1.0
154.8 ± 1.0
105.1 ± 0.9
DBP, mm Hg
58.3 ± 0.3
101.3 ± 0.3
HR, beats/min
61.5 ± 0.8
BMI, kg/m^2
METABOLIC
Total cholesterol,
mg/dl
HDL cholesterol,
mg/dl
N
PHYSICAL
RENAL
Serum creatinine,
mg/dl
On-line Table II. Phenotype (CHGA and Catecholamine) and BP Study: Subject Characteristics.
724 individuals of diverse ancestries were phenotyped for not only blood pressure, but also indices of
sympathochromaffin activity (plasma norepinephrine and CHGA concentrations), as well as metabolic
and renal traits. Hypertension was defined in Methods, and was treated with antihypertensive medications
in 75% of such subjects.
Sex
2-Way ANOVA
Male
BP status
Female
BP Status
Sex
BP-by-Sex
p Value
P Value
p Value
Normotensive
Hypertensive
Normotensive
Hypertensive
174
24
470
56
SBP, mm Hg
133.7 ± 1.1
157.2 ± 2.8
127.4 ± 0.7
145.8 ± 2.1
<0.001
<0.001
0.175
DBP, mm Hg
73.0 ± 0.8
89.3 ± 1.89
70.08 ± 0.46
78.6 ± 1.4
<0.001
<0.001
0.002
HR, beats/min
68.7 ± 0.9
70.5 ± 2.2
70.5 ± 0.5
71.6 ± 1.7
0.711
0.102
0.697
BMI, kg/m^2
25.9 ± 0.5
29.3 ± 1.3
24.3 ± 0.3
29.6 ± 0.8
<0.001
0.341
0.044
SYMPATHETIC
Plasma CHGA[116-439],
nM
Plasma norepinephrine,
pg/ml
3.93 ± 0.28
8.07 ± 0.52
3.84 ± 0.17
4.38 ± 0.51
<0.001
<0.001
0.001
342.8 ± 12.1
474.3 ± 32.7
323.9 ± 7.3
367.8 ± 23.0
<0.001
0.002
0.033
0.955 ± 0.02
0.982 ± 0.04
0.773 ± 0.01
0.762 ± 0.03
0.757
<0.001
0.458
Total cholesterol, mg/dl
164.0 ± 4.4
177.0 ± 9.7
178.5 ± 2.25
190.2 ± 7.26
0.060
0.035
0.922
HDL cholesterol, mg/dl
40.1 ± 1.8
43.8 ± 4.0
54.72 ± 0.92
50.17 ± 2.97
0.873
<0.001
0.124
107.7 ± 8.58
126.1 ± 18.9
103.7 ± 4.40
170.1 ± 14.2
0.001
0.118
0.061
N
PHYSICAL
RENAL
Serum creatinine, mg/dl
METABOLIC
Triglycerides, mg/dl