新生兒耳聾基因篩檢
• 輕度聽損
• 遲發性聽損
新生兒耳聾基因篩檢，可輔助傳統新生兒聽力篩檢，增加檢出率
我們所進行的新生兒耳聾基因篩檢，初步結果顯示我們現在所採行之新生兒聽力 篩檢流程，有可能遺漏較輕度或晚發性之兒童聽損，而此一部分，似可以耳聾基 因篩檢加以補足。
圖四十
Endolymph Stria vascularis Epithelial cell
Pendrin
HCO3 -HCO3
-Cl
-H+ H+
SLC26A4 mutation
pH PP
PPI
Free radical
stress
Predisposition to hearing loss
質離子幫浦阻斷劑(PPI)作用於 SLC26A4 基因突變病人急性聽損之可能機制 SLC26A4 基因所製造的蛋白質—pendrin，在內耳之功能為將重碳酸根離子(HCO3-) 從血管紋(stria vascularis)運送至內淋巴，故若發生突變，將導致血管紋內 pH 值 升高，並引起自由基累積及急性聽損。而質離子幫浦於血管紋之功能，則為將質 子從血管紋運送至內淋巴，因此，在質離子幫浦阻斷劑的作用下，恰可防止血管 紋內pH 值的升高。
表一、420 個家族初始受試者之基本資料
Variables
Total probands
(n=420)
Probands from hospitals
(n=325)
Probands from rehabilitation
facilities (n=95) Age (yr, median [range]) 8 [1-57] 10 [1-57] a 5 [1-20] a Gender (M : F) 215 : 205 167 : 158 b 48 : 47 b Familial : Sporadic 114 : 306 95 : 230 b 19 : 76 b Clinical diagnosis
Syndromic 29 22 c 7 c
Non-syndromic 391 303 88 Without IEMs 288 217 c 71 c
With EVA or MD 85 73 c 12 c
With other IEMs 18 13 c 5 c
Cochlear implantation
Yes : No 89 : 331 58 : 267 d 31 : 64 d IEM = inner ear malformation; EVA = enlarged vestibular aqueduct; MD = Mondini’s dysplasia (incomplete partition of cochlea).
a Mann-Whitney U test, p < 0.01.
b Chi-square test, p > 0.05.
c Chi-square test for heterogeneity in distribution of clinical diagnosis, df = 3, p >
0.05.
d Chi-square test, p < 0.05.
表二、於 420 個家族中三段常見耳聾基因之基因變異光譜
Exon Nucleotide change Codon
change
No. of variant alleles SLC26A4
3 c.230A>T p.K77I 1
3’ Int 7 c.919-2A>G splice acceptor 102 5’ Int 8 c.1001+5G>C splice donor 1
9 c.1115C>T p.A372V 3
10 c.1160C>T p.A387V 2
10 c.1229C>T p.T410M 3
12 c.1343C>T p.S448L 3
13 c.1489G>C p.G497R 1
19 c.2162C>T p.T721M 1
19 c.2168A>G p.H723R 4
GJB2
2 c.109G>A* p.V37I* 129
2 c.124G>A p.E42K 2
2 c.187G>T p.V63L 1
2 c.230G>A p.W77X 1
2 c.235delC frameshift 38
2 c.299_300delAT frameshift 8
2 c.427C>T p.R143W 1
2 c.508_509insAACG frameshift 1
2 c.571T>C p.F191L 1
Mito. 12S rRNA
- m.1555A>G - 12
- m.961delT+C(n) - 4
* Pathogenicity unclear.
表三、醫院(Group A)及聽語復健中心(Group B)初始受試者中常見耳聾基因變異對偶基因頻率的差異
Genes/Variant alleles
Total
Variant alleles no.
(%) in the 420 probandsa
Variant alleles no.
(%) in Group Ab
Variant alleles no.
(%) in Group Bc
Comparison between group
A & B
a 840 GJB2 and SLC26A4 alleles, but 420 mitochondria 12S rRNA alleles.
b 650 GJB2 and SLC26A4 alleles, but 325 mitochondria 12S rRNA alleles.
c 190 GJB2 and SLC26A4 alleles, but 95 mitochondria 12S rRNA alleles.
d Chi-square test.
e Fisher’s exact test.
表四、醫院(Group A)及聽語復健中心(Group B)初始受試者中 SLC26A4 基因 型分佈的比較
Genotypes
Probands of group A
(n=325)
no. (%)
Probands of group B
(n=95)
no. (%)
2 variant alleles detected 40 (12 %)a 7 (7%)a
1 variant allele detected 25 (8%)a 2 (2%)a
No variant detected 260 (80%)a 86 (91%)a
a Chi-square test for heterogeneity in distribution of genotype prevalence, df = 2, p
< 0.05.
表五、醫院(Group A)及聽語復健中心(Group B)初始受試者中聽力學特徵之比 較
Variable
Probands of group A (n=325)
no. (%)
Probands of group B Hearing loss patterns
Fluctuating 67 (21%) 13 (14%) 80 (19%) Progressive 108 (33%) 10 (11%) 118 (28%) Stationary 150 (46%) 72 (76%) 222 (53%)
― p < 0.01 a Audiogram configurations
Sloping 196 (60%) 46 (48%) 242 (58%) Flat type 120 (37%) 46 (48%) 166 (40%) Low tone 3 (1%) 1 (1%) 4 (1%)
Others 6 (2%) 2 (2%) 8 (2%)
― p > 0.05 b Hearing levels
Mild (20~40dBHL) 23 (7%) 4 (4%) 27 (6%) Moderate (41~70 dBHL) 108 (33%) 16 (17%) 124 (30%) Severe (71~95 dBHL) 94 (29%) 33 (35%) 127 (30%) Profound (> 95 dBHL) 100 (31%) 42 (44%) 142 (34%)
― p < 0.01 c
a Chi-square test for heterogeneity in distribution of hearing loss patterns between probands from hospitals and those from rehabilitation facilities, df = 2, p < 0.01.
b Chi-square test for heterogeneity in distribution of audiogram configurations between probands from hospitals and those from rehabilitation facilities, df = 3, p
> 0.05.
c Chi-square test for heterogeneity in distribution of hearing levels between probands from hospitals and those from rehabilitation facilities, df = 3, p < 0.01.
表六、以羅吉斯迴歸分析聽力學特徵與 SLC26A4 基因型之相關性 Variable Adjusted OR 95% CI p-value a Age 1.01b 0.97 – 1.05 0.580 Gender (male) 1.01 0.45 – 2.23 0.988 Hearing loss pattern
Stationary 1.00
Fluctuating 148.83 50.84 – 435.72 < 0.001 Progressive 4.51 1.31 – 15.55 0.017 Audiogram configuration
Non-sloping 1.00
Sloping 2.49 1.05 – 5.89 0.039 Hearing level
Mild + Moderate 1.00
Severe + Profound 3.92 1.62 – 9.49 0.003 OR = odds ratio; CI = confidence interval.
a Wald test.
b Odds ratio for per one-year change.
表七、 於 101 個 Pendred 氏症候群和非症候群型大前庭導水管家族之 SLC26A4 基因型分布
No. (%) of families by No. of SLC26A4 mutations
Zero One Two Multiplex families (n=32) 3 4 25
Simplex families (n=69) 11 20 38
Total (n=101) 14 24 63
Multiplex families, more than one familial member affected； simplex families, only one familial member affected.
表八、於 101 個 Pendred 氏症候群和非症候群型大前庭導水管家族之 SLC26A4 基因變異
Exon Nucleotide change Codon change No. of variant alleles*
3 c.230A>T p.K77I 1
3 c.235C>T p.R79X 1
3 c.281C>T p.T94I 2
7 c.916_917insG p.V306GfsX24 1
3’ Int 7 c.919-2A>G splice acceptor 115 5’ Int 8 c.1001+5G>C splice donor 1
9 c.1115C>T p.A372V 4
10 c.1160C>T p.A387V 2
10 c.1174A>T p.N392Y 1
10 c.1225C>T p.R409C 1
10 c.1229C>T p.T410M 4
12 c.1343C>T p.S448L 3
13 c.1489G>C p.G497R 1
5’ Int 15 c.1705+5G>A splice donor 1
16 c.1786C>T p.Q596X 1
19 c.2162C>T p.T721M 1
19 c.2168A>G p.H723R 10
Total 150
* Among the 202 SLC26A4 alleles of the 101 probands.
表九、 於 c.919-2A>G 同型合子、異型合子及對照組所進行之 SNP 分析 JST160568
Subjects CC CT TT P-value
c.919-2A>G x2 (n=16) 0 0 16a P < .001 a c.919-2A>G x1 (n=16) 0 3 13
Controls (n=50) 3 19 28a JST089508
Subjects CC CA AA P-value
c.919-2A>G x2 (n=16) 16b 0 0 P < .0001 b c.919-2A>G x1 (n=16) 8 8 0
Controls (n=50) 8b 22 20 JST160566
Subjects CC CG GG P-value
c.919-2A>G x2 (n=16) 15c 1 0 P < .0001 c c.919-2A>G x1 (n=16) 7 9 0
Controls (n=50) 4c 20 26 JST160565
Subjects GG GC CC P-value
c.919-2A>G x2 (n=16) 14d 2 0 P < .0001 d c.919-2A>G x1 (n=16) 5 7 4
Controls (n=50) 6d 23 21
a Rows combined for Fisher’s exact test; b Rows combined for Fisher’s exact test;
c Rows combined for Fisher’s exact test; d Rows combined for Fisher’s exact test
表十、SLC26A4 基因型與表現型之關連性 Probands with 2 alleles
of truncating mutations
(n=39)
Other probands with 2 mutated
alleles (n=24)
Probands with 1 mutated
allele (n=24)
Probands with 0 mutated
allele (n=14)
p-value*
Types of IEMs, n (%)
Isolated EVA 15 (38) 12 (50) 10 (42) 8 (57) 0.60 EVA + other IEMs 24 (62) 12 (50) 14 (58) 6 (43)
Goiter, n (%) 3 (8) 2 (8) 2 (8) 0 (0) 0.86 Hearing levels, dBHL, mean (SD) 86.2 (17.7) 87.4 (17.1) 82.0 (19.0) 81.1 (23.9) 0.63 Fluctuating hearing loss, n (%) 33 (85) 19 (79) 16 (67) 7 (50) 0.06 Sloping audiogram, n (%) 29 (74) 17 (71) 18 (75) 7 (50) 0.36
IEM = inner ear malformation; EVA = enlarged vestibular aqueduct.
* p-value by χ2 or Fisher’s exact test for categorical variables and ANOVA for continuous variables.
表十一、SLC26A4 基因促進區(promoter)及 FOXI1 基因之基因變異
Gene/Region Nucleotide change Codon change No. (%) of alleles*
SLC26A4 promoter
Region 1 (c.-2988 to c.-2258) c.-2554G>A - 1 (1) Region 2 (c.-1063 to c.-325) c.-964A>C - 19 (25) FOXI1
Exon 1 c.279G>A p.R93R 35 (46)
* Among the 76 alleles of the 38 probands with 1 or 0 mutated SLC26A4 allele detected.
表十二、21 名 c.919-2A>G 異型合子病人之未被偵測出突變 SLC26A4 對偶基因之單套型 Haplotype
rs#2248464 rs#2248465 rs#2712212 rs#3839817 rs#2395911 rs#2072064 rs#2072065 rs#2301634
No. (%) of alleles (n=21)
A C T TCTTT A T T T 4 (19)
A C T TCTTT C A C T 4 (19)
T C C TCTTT C A C T 1 (5)
T C T TCTTT A T T T 3 (14)
T C T TCTTT C A C T 1 (5)
T T C TCTTT C A C T 1 (5)
T T C - A T T T 1 (5)
T T T TCTTT A T T T 3 (14)
T T T TCTTT C A C T 3 (14)
196
表十三、不同組間基因變異對偶基因頻率之比較
Allele no. (%) Genes/Variant alleles Severe-to-
profound SNHI a
Mild-to- moderate
SNHI b
Control c
GJB2
p.V37I 91 (12.8) d 228 (30.3) e 184 (9.2) Other definite mutations 48 (6.7) 28 (3.7) 5 (0.2) SLC26A4
Definite mutations 144 (20.2) 48 (6.4) NA Mito. 12S rRNA
m.1555A>G 16 (4.5) 15 (4.0) NA
a 712 GJB2 and SLC26A4 alleles, but 356 mitochondria 12S rRNA alleles.
b 752 GJB2 and SLC26A4 alleles, but 376 mitochondria 12S rRNA alleles.
c 2010 GJB2 alleles.
d Chi-square test, p = 0.007 as compared to the control group.
e Chi-square test, p < 0.001 as compared to the control group.
NA = not available
197
表十四、聽損程度不同組間 p.V37I 變異基因型分布之比較 Genotype (n)
p.V37I/p.V37I p.V37I/others others/others
p-value
Mild-to-moderate SNHI (n=376)
Observed no. 77 74 225 p < 0.001*
Expected no.** 3.2 62.8 310
Severe-to-profound SNHI (n=356)
Observed no. 9 73 274 p < 0.001*
Expected no. 3.0 59.5 293.5
* Chi-square goodness-of-fit test.
** Assumed allele frequency of p.V37I = 0.092
表十五、不同 p.V37I 基因型病人基本資料與聽力表徵之比較
Variable p.V37I/p.V37I
(n=86)
p.V37I/MT*
(n=12)
Others
(n=634) p-value
Male, n (%) 52 (60) 6 (50) 295 (47) 0.55**
Age, median [range], y 16 [1-54] 18 [2-42] 12.5 [1-75] 0.61***
Hearing levels, mean (SD), dBHL 48.5 (18.7) 58.5 (27.0) 73.2 (25.9) < 0.001****
Hearing loss patterns, n (%)
Fluctuating 4 (5) 0 (0) 113 (18) 0.11**
Progressive 49 (57) 6 (50) 218 (34)
Stationary 33 (38) 6 (50) 303 (48)
Audiogram configurations, n (%)
Sloping 49 (57) 8 (67) 330 (52) 0.40**
Flat type 29 (34) 4 (33) 254 (40)
High tone 7 (8) 0 (0) 18 (3)
Others 1 (1) 0 (0) 32 (5)
* MT indicates other GJB2 mutations, including 9 c.235delC mutation and 3 c.299_300delAT mutation.
** P-value by χ2 or Fisher’s exact test.
*** Wilcoxon rank-sum test.
***** ANOVA, post hoc test for significance with the Tukey multiple comparison procedure: genotype p.V37I/p.V37I vs others, p < 0.001.
199
表十六、1005 名受試者基本資料 (mean (S.D.) or n (%))
Variables Value
Age (years) 56.4 (9.0)
Male sex (%) 487 (48.5)
Smoking (%) 137 (13.6)
Alcohol consumption (%) 292 (29.1) Coronary heart disease (%) 49 (4.9) Cerebral vascular disease (%) 5 (0.5) Hypertension (%) 188 (18.7)
Diabetes (%) 82 (8.2)
Hyperlipidemia (%) 84 (8.4) Renal disease (%) 14 (1.4)