Genome-wide association study reveals HSF2, GJA1 and TRIM36 as susceptibility genes for preeclampsia: a community-based population study in Tianjin, China

Figures & data

Figure 1. Flow chart of the study subject recruitment process.

Table 1. Summary of patient characteristics in discovery cohort.

	case(n=92)	control(n=187)	p
Age	31.25 ± 3.87	31.62 ± 3.79	0.45
Height (cm)	162.65 ± 5.95	163.13 ± 4.93	0.48
Weight (kg)	65.58 ± 13.95	63.05 ± 11.10	0.13
Gravidity	1.65 ± 1.02	1.84 ± 1.10	0.16
Parity	0.20 ± 0.43	0.29 ± 0.47	0.09
Initial systolic blood pressure (mmHg)	113.89 ± 15.29	111.91 ± 10.82	0.27
Initial diastolic blood pressure (mmHg)	72.75 ± 9.96	70.51 ± 8.50	0.07
Red blood cell (×10^12/L)	4.43 ± 0.55	4.25 ± 0.35	0.00
White blood cell (×10^9/L)	8.60 ± 2.04	8.24 ± 1.86	0.15
Hemoglobin (g/L)	128.64 ± 11.69	126.62 ± 9.94	0.13
Platelet (×10^9/L)	247.48 ± 61.89	244.27 ± 59.16	0.68
Fasting blood glucose (mmol/L)	4.91 ± 0.64	4.54 ± 0.80	0.00
Total protein (g/L)	69.75 ± 3.73	67.41 ± 3.79	0.00
Albumin (g/L)	43.03 ± 3.16	42.02 ± 2.47	0.03
Alanine aminotransferase	27.50 ± 28.75	23.01 ± 14.83	0.16
Urea nitrogen (mmol/L)	3.17 ± 0.70	3.25 ± 2.42	0.74
Creatinine (μmol/L)	58.35 ± 11.40	63.38 ± 16.08	0.00
Uric acid (μmol/L)	229.16 ± 61.96	218.34 ± 52.79	0.13
Gestational age of delivery	38.31 ± 2.14	39.39 ± 1.25	0.00
Weight before delivery	77.28 ± 13.46	75.71 ± 11.90	0.33
Neonatal weight	3008.02 ± 766.22	3260.58 ± 507.94	0.01
Length of newborn	48.04 ± 6.01	49.57 ± 1.95	0.02
42 days postpartum systolic blood pressure (mmHg)	117.34 ± 14.16	113.63 ± 10.14	0.04
42 days postpartum diastolic blood pressure (mmHg)	75.99 ± 11.39	74.65 ± 8.17	0.35

Table 2. Summary of patient characteristics in validation cohort.

	case(n=52)	control(n=104)	p
Age	30.83 ± 3.99	30.73 ± 3.34	0.87
Height(cm)	163.39 ± 5.49	161.82 ± 16.60	0.51
Weight(kg)	64.55 ± 11.27	63.13 ± 10.12	0.43
Gravidity	1.54 ± 0.92	1.93 ± 0.96	0.02
Parity	0.12 ± 0.32	0.43 ± 0.54	0.00
Initial systolic blood pressure(mmHg)	111.62 ± 14.48	105.46 ± 9.31	0.01
Initial diastolic blood pressure(mmHg)	72.69 ± 9.67	66.56 ± 7.34	0.00
Red blood cell (×10^12/L)	4.29 ± 0.44	4.24 ± 0.35	0.53
White blood cell (×10^9/L)	8.06 ± 1.57	7.76 ± 1.67	0.28
Hemoglobin (g/L)	129.81 ± 11.26	127.80 ± 10.49	0.27
Platelet (×10^9/L)	231.63 ± 64.84	229.34 ± 52.19	0.81
Fasting blood glucose (mmol/L)	4.95 ± 0.65	4.80 ± 0.44	0.10
Total protein (g/L)	69.23 ± 3.73	69.90 ± 4.60	0.48
Albumin (g/L)	43.79 ± 2.38	44.09 ± 3.16	0.64
Alanine aminotransferase	28.65 ± 29.54	21.74 ± 26.62	0.14
Urea nitrogen (mmol/L)	2.95 ± 0.57	3.03 ± 0.62	0.47
Creatinine (μmol/L)	53.19 ± 10.62	51.46 ± 11.93	0.38
Uric acid(μmol/L)	219.48 ± 48.97	210.04 ± 50.08	0.27
Gestational age of delivery	38.31 ± 2.14	39.39 ± 1.25	0.00
Weight before delivery	75.88 ± 10.46	77.57 ± 22.10	0.60
Neonatal weight	3055.77 ± 606.84	3418.21 ± 443.57	0.00
Length of newborn	48.21 ± 7.07	50.27 ± 1.89	0.05
42 days postpartum systolic blood pressure(mmHg)	115.24 ± 10.19	109.40 ± 10.04	0.00
42 days postpartum diastolic blood pressure (mmHg)	74.72 ± 7.81	70.48 ± 7.42	0.00

Figure 2. Q‒Q plots with P values under the expected zero distribution relative to theoretical P values.

Figure 3. Manhattan chart at the preliminary GWAS discovery stage of severe preeclampsia.

Figure 4. GO bubble analysis regarding preeclampsia-related pathways.

Table 3. The initial GWAS result in the discovery stage.

CHR	dbSNPID	POSITION	gene	gene_relationship	A1	A2	MAF		OR	P (fisher)
							case	control
13	rs9523993	93930403	GPC6	genic	G	A	0.40	0.23	2.219	3.59×10^−5
2	rs886043316	20130280	WDR35	genic	A	C	0.10	0.02	5.216	7.07×10^−5
2	rs78613705	163507406	KCNH7	genic	A	G	0.19	0.07	3.165	2.69×10^−5
1	rs77356340	245842272	KIF26B	genic	A	G	0.09	0.01	7.052	4.42×10^−5
2	rs76289759	163460529	KCNH7	genic	A	G	0.27	0.13	2.575	3.30×10^−5
13	rs75154137	95626009	LINC00557	intergenic	T	C	0.09	0.01	7.604	1.61×10^−5
20	rs73299501	56168784	ZBP1,PCK1	intergenic	A	G	0.15	0.05	3.622	3.61×10^−5
12	rs73095909	47679298	PCED1B,MIR4494	intergenic	G	A	0.13	0.03	4.41	3.13×10^−5
10	rs7068679	130318498	LINC01163,MGMT	intergenic	A	G	0.13	0.03	4.171	6.99×10^−5
17	rs56309431	63645289	CEP112	genic	T	G	0.60	0.42	2.108	4.07×10^−5
14	rs148761074	104502811	TDRD9	genic	T	C	0.08	0.01	10.67	1.49×10^−5
1	rs147968844	30829270	LOC101929406,MATN1	intergenic	T	C	0.06	0.00	NA	4.82×10^−5
3	rs146895271	57582163	ARF4	genic	G	T	0.11	0.02	5.34	6.15×10^−5
2	rs146718393	137647360	THSD7B	genic	A	G	0.09	0.01	7.105	4.07×10^−5
6	rs13210237	122346667	HSF2,GJA1	intergenic	A	G	0.24	0.10	2.793	2.54×10^−5
5	rs13176432	114369723	TRIM36	intergenic	G	T	0.11	0.02	4.85	6.67×10^−5
8	rs117487811	1647942	DLGAP2	genic	A	C	0.09	0.02	5.576	9.11×10^−5
16	rs117222557	11147978	CLEC16A	genic	A	G	0.10	0.02	5.642	8.75×10^−5
7	rs10229442	146503040	CNTNAP2	genic	T	C	0.05	0.00	NA	4.76×10^−5

Table 4. SNP screened from validation cohort.

					MAF					95% Confidence Interval
chr	SNP	BP	A1	A2	case	control	CHISQ	P	OR	Lower	Upper
13	rs9523993	93930403	G	A	0.28	0.27	0.01	0.929	1.02	0.61	1.73
2	rs886043316	20130280	C	C	0.00	0.00			NS
2	rs78613705	163507406	A	G	0.07	0.13	2.44	0.118	0.51	0.21	1.21
1	rs77356340	245842272	A	G	0.01	0.02		0.668	0.39	0.05	3.42
2	rs76289759	163460529	A	G	0.13	0.19	1.95	0.163	0.62	0.31	1.22
13	rs75154137	95626009	T	C	0.01	0.03	1.17	0.431	0.33	0.04	2.75
20	rs73299501	56168784	A	G	0.05	0.04	0.16	0.766	1.26	0.40	3.96
12	rs73095909	47679298	G	A	0.04	0.05	0.32	0.575	0.72	0.22	2.31
10	rs7068679	130318498	A	G	0.06	0.03	1.00	0.371	1.76	0.58	5.37
17	rs56309431	63645289	T	G	0.50	0.51		0.936	0.98	0.61	1.57
14	rs148761074	104502811	T	C	0.02	0.04		0.505	0.49	0.10	2.35
1	rs147968844	30829270	T	C	0.01	0.03		0.431	0.33	0.04	2.75
3	rs146895271	57582163	G	T	1.00	0.99		0.553	0.00
2	rs146718393	137647360	A	G	0.01	0.01		1.000	0.66	0.07	6.46
6	rs13210237	122346667	A	G	0.16	0.09	4.12	0.042	2.06	1.01	4.19
5	rs13176432	114369723	G	T	0.07	0.02		0.047	0.27	0.08	0.95
8	rs117487811	1647942	A	C	0.04	0.04		1.000	0.88	0.27	2.94
16	rs117222557	11147978	A	G	0.05	0.06	0.12	0.724	0.83	0.28	2.41
7	rs10229442	146503040	T	C	0.02	0.01		0.259	4.06	0.36	45.29

Table 5. Consolidation calculation of two stages.

Chr	SNP	BP	A1	A2	case (n=144)	control (n=291)	CHISQ	P	OR	95% Confidence Interval
										Lower	Upper
5	rs13176432	114369723	G	T	0.090	0.022		5×10^−6	2.054	1.315	3.209
6	rs13210237	122346667	A	G	0.205	0.096	19.823	8×10^−6	1.431	1.179	1.736

Table 6. Comparison of genotypes between early-onset and late-onset preeclampsia.

Genotype and model	Early-onset (n=33)	Late-onset (n=111)	χ2	P
rs13176432
GG	0	2	Fisher exact	.872
TG	4	18
TT	29	91
Dominant model GG + TG vs TT			0.6369	.425
GG+TG	4	20
TT	29	91
Recessive model GG vs TT+ TG			Fisher exact	>.999
GG	0	2
TT+TG	33	109
rs13210237
AA	1	4	Fisher exact	.142
GA	16	33
GG	16	72
Dominant model AA + GA vs GG			3.318	.068
AA+GA	17	37
GG	16	72
Recessive model AA vs GA + GG			χ2Yates=0.132	.716
AA	1	4
GA+GG	32	105

Table 7. The demographic and clinical characteristics of PE patients and controls.

	PE		non-PE		χ2 / Z	P
	N=144		N=291
Maternal age(year)	31	(29,33)	30	(29,34)	-0.284	0.776
height(cm)	162	(159,166)	163	(160,166)	-0.977	0.329
BMI (Kg/m^2)	24.06	(21.34,28.51)	21.02	(19.13,24.23)	2.563	0.000
Weight gain(kg)	16	(11.1,20)	15	(11,20)	-1.379	0.168
Diastolic BP (mm Hg)	100	(90,120)	70	(60,70)	-4.457	0.000
Systolic BP (mm Hg)	158	(130,180)	100	(100,110)	-5.167	0.000
diabetes [n(%)]	41	(28.47)	37	(12.71)	16.254	0.000
Multiple pregnancy [n(%)]	9	(6.25)	5	(1.72)	4.980	0.026
gravidity≥2[n(%)]	56	(38.89)	143	(49.14)	4.079	0.043
History of miscarriage [n(%)]	41	(28.47)	91	(31.27)	0.357	0.550
family history of hypertension [n(%)]	39	(27.08)	82	(28.18)	0.058	0.810
in vitro fertilization [n(%)]	8	(5.56)	16	(5.50)	0.001	0.980
Education background
≤9 Year (%)	11	(7.64)	20	(6.87)
9-12 Year (%)	16	(11.11)	27	(9.28)	-0.007	0.994
12-16 Year (%)	100	(69.44)	214	(73.54)
>16 Year (%)	17	(11.81)	30	(10.31)
Monthly household income (RMB)
<5000/case (%)	88	(61.11)	64	(21.99)
5000-10000/case (%)	138	(95.83)	55	(18.90)	-2.662	0.008
≥10000/case(%)	65	(45.14)	25	(8.59)
rs13176432 Dominant model [n(%)]	24	(16.67)	13	(4.47)	18.422	0.000
rs13176432 Recessive model [n(%)]	2	(1.39)	0	(0.00)	1.593	0.207
rs13210237 Dominant model [n(%)]	54	(37.50)	54	(18.56)	18.522	0.000
rs13210237 Recessive model [n(%)]	5	(3.47)	2	(0.69)	3.124	0.077
red blood cell (×10^12/L)	4.21	(3.755,4.5)	4.1	(3.7, 4.36)	-2.015	0.044
White blood cell (×10^9/L)	8.685	(7.325,9.975)	7.8	(6.4, 9.2)	-3.969	0.000
hemoglobin (g/L)	131	(124,136)	128	(122,135)	-2.013	0.044
platelet (×10^9/L)	245	(201.25,275.7)	218	(184,256)	-3.398	0.001
Fasting blood glucose (mmol/L)	4.895	(4.56,5.26)	4.84	(4.56,5.1)	-1.554	0.120
Alanine aminotransferase (U/L)	18.2	(13.15,26.37)	16	(11,24)	-2.632	0.008
urea nitrogen (mmol/L)	3	(2.575,3.5575)	3.02	(2.6,3.37)	-0.382	0.702
creatinine (μmol/L)	52	(46,61.2625)	49.2	(44,57.6)	-1.968	0.049
uric acid(μmol/L)	222.55	(190.25,268.8)	203.4	(171.8,232.1)	-3.99	0.000

The results showed that BMI before pregnancy, systolic blood pressure, gestational diabetes, multiple pregnancy, pregnancy times ≥ 2, rs13210237 dominant mode, rs13176432 dominant mode, platelet count in early pregnancy may be related to preeclampsia.

Table 8. Multiple logistic regression analysis.

						95% CI
	B	S.E.	Wald	P	Exp(B)	Lower	Upper
BMI	0.097	0.031	9.687	0.002	1.102	1.037	1.172
Diastolic BP	0.036	0.011	11.223	0.001	1.037	1.015	1.059
Diabetes mellitus	0.752	0.291	6.696	0.01	2.121
Multiple pregnancy	1.821	0.65	7.846	0.005	6.177	1.728	22.083
Gravidity≥2	−0.526	0.241	4.774	0.029	0.591	0.369	0.947
Rs13210237 Dominant model	1.144	0.263	18.871	0.000	3.138	1.873	5.256
Rs13176432 Dominant model	1.769	0.403	19.254	0.000	5.867	2.662	12.932
Platelet	0.006	0.002	9.496	0.002	1.006	1.002	1.009
	−10.492	1.499	49.008	0	0