JAG1 Variants Confer Genetic Susceptibility to Thyroid Dysgenesis and Thyroid Dyshormonogenesis in 813 Congenital Hypothyroidism in China

Figures & data

Table 1 The Clinical Characteristics of All CH Subjects and JAG1 Variant Carriers

		All Subjects	JAG1 Variant Carriers	p value
Gender (males/females)(n)		393/420	12/13	0.973
Birth weight, median (IQR) (kg)		3.4(3.1–3.7)	3.5(3.1–3.6)	0.787
Diagnostic serum	TSH level, median (IQR) (mIU/L)	>75(44.3–75)	>75(47.5–79)	0.558
	T3 level, median (IQR) (nmol/L)	1.7(1.0–2.3)	1.6(1.0–2.2)	0.560
	T4 level, median (IQR) (nmol/L)	34.4(12.9–66.3)	26.5(14.45–59.7)	0.653
	fT3 level, median (IQR) (pmol/L)	5.0(3.2–6.0)	4.2(3.56–5.7)	0.616
	fT4 level, median (IQR) (pmol/L)	6.2(3.9–9.6)	4.7(3.0–7.1)	0.043
CH severity (N, percentage)	severe (<5 pmol/L)	333, 41.0%	14, 56%	0.306
	moderate (5 to 10 pmol/L)	268, 33.0%	8, 32.0%
	mild (>10 pmol/L)	181, 22.3%	3, 12.0%
Pathogenesis (N, percentage)	thyroid dysgenesis	224, 27.6%	8, 32.0%	0.287
	thyroid dyshormonogenesis	589, 72.4%	17, 68.0%
CH phenotype (N, percentage)	PCH	296,	12,	0.183
	TCH	211,	4,
	unclassified	306	9
Other clinical features (N)	Prolonged jaundice	149	3
	Abdominal distension	81	5
	Umbilical hernia	40	2
	Low heart sounds	32	2
	Hoarse cry	21	1
	Drowsiness	9	0
	Cardiac murmurs	4	0
	Protruding tongue	1	0
	High muscular tension in both lower limbs	1	0

Notes: Mann Whitney U-test and chi-square test were used for statistical analysis.

Abbreviations: IQR, interquartile range; PCH, permanent congenital hypothyroidism; TCH, transient congenital hypothyroidism; NBS, newborn screening; L-T4, L-thyroxine.

Table 2 Clinical Characteristics of Patients with Potential Pathological Variants

Patient	Sex	Diagnostic Evaluation					CH Severity	CH Phenotype	Thyroid Morphology	Other Clinical Features	Variant 1 in JAG1	Variant 2	Variant 3
		FT3	FT4	T3	T4	TSH
		pmol/L		nmol/L		mIU/L
1	M	9.17	6.5	2.65	37.7	74.5	moderate	PCH	Agenesis		p.V45L
2	M	3.65	12	1.8	110	37.7	mild	NA	Normal
3	F	5.4	4.39	1.4	20.8	>75	severe	NA	Normal	Prolonged jaundice Abdominal distension
4	F	1.9	<3.86	<0.62	<12.9	>75	severe	PCH	Hypoplasia	Prolonged jaundice Abdominal distension Umbilical hernia Low heart sounds Hoarse cry
5	M	6	5.1	1.8	22.3	50.3	moderate	TCH	Goiter
6	F	3.9	0.42	2.21	2.43	83	severe	PCH	Normal			DUOX2 p.R1485G
7	F	2.5	10.3	0.6	36	50.3	mild	NA	Goiter			DUOX2 p.R1110Q
8	M	4.6	4.71	1.6	17.2	>75	severe	NA	Goiter	Abdominal distension		DUOX2 p.R1110Q	DUOX2 p.V407F
9	M	5.5	6.34	1.7	32.6	>75	moderate	NA	Normal		p.V272I
10	M	5	6.24	2.17	27.2	100	moderate	PCH	Normal		p.P552L
11	F	1.4	1.61	0.5	20	100	severe	PCH	Agenesis
12	M	3.47	3.86	1.04	14.5	100	severe	PCH	Hypoplasia
13	M	7.3	7.26	4.3	59.3	>75	moderate	NA	Goiter
14	M	<1	<3	<0.4	<10	>75	severe	TCH	Normal	Prolonged jaundice		DUOXA2 p.I26M
15	M	3.76	1.27	1.43	89.4	12.75	severe	NA	Normal		p.G610E
16	F	8.7	0.05	2.2	180	21.8	severe	PCH	Goiter			DUOX2 p.R1110Q
17	F	3.8	4.7	1.2	20.8	>75	severe	PCH	Agenesis		p.G852D
18	M	4.1	9.2	1.1	61.8	44.6	moderate	PCH	Hypoplasia		p.A891T
19	F	5.4	7.16	2.5	52.5	39.7	moderate	TCH	Normal	Abdominal distension Umbilical hernia	p.E1030K
20	M	6.99	5	2	26.5	100	severe	NA	Normal		p.R1060W	DUOX2 p.P76L	DUOX2 p.R625X
21	F	1.8	3	<0.4	<10	>75	severe	PCH	Agenesis	Abdominal distension Low heart sounds	p.A1131T
22	F	5.9	7.6	2.4	60.1	42.7	moderate	TCH	Normal
23	F	4.7	<7	2.23	66.7	100	moderate	PCH	Agenesis			DUOX2 p.I967T
24	F	3.8	<3.86	1	<12.9	>75	severe	NA	Goiter		p.P1174L	DUOX2 p.R1110Q
25	F	4.2	<3	1.3	14.4	>75	severe	PCH	Goiter				DUOX2 p.R82C

Abbreviations: PCH, permanent congenital hypothyroidism; TCH, transient congenital hypothyroidism.

Figure 1 Multiple sequences alignment. Multiple sequences alignment were conducted using ClustalW2 among Homo sapiens, Mus musculus, Rattus norvegicus, Sus scrofa, Pan troglodytes, Bos Taurus, Macaca mulatta and Felis catus. (A) Multiple sequences alignment of JAG1; (B) Multiple sequences alignment of DUOX2; (C) Multiple sequences alignment of DUOXA2. The mutation sites were marked by red arrows.

Figure 2 The changes of hydrophobic parameters caused by amino acid substitution. Wild-type amino acids were marked with black, substituted amino acids were marked with red; “+” means increased hydrophobicity, “-” means decreased hydrophobicity.

Figure 3 Analysis of genotype and phenotype relationship (A) The distribution of the number of variants carried by each patients; (B) The comparison of fT4 concentration between monogenic group and oligogenic group; (C) The comparison of fT4 concentration between monogenic DH and oligogenic DH; (D) The comparison of fT4 concentration between monogenic TD and monogenic DH. “ns” means P>0.05, “*” means P<0.05.

Figure 4 Clustering analysis of JAG1 variants in CH. Distance in nucleotides between missense mutations within exons 1–21 and exons 22–26. Statistical significance was calculated using an unpaired, two-tailed t test, “*” means P<0.05.