Edward Mallinckrodt Department of Pediatrics, Washington University School of Medicine, 660 South Euclid, St. Louis, MO 63110, USA.
Pediatr Res. 2010 Sep;68(3):216-20. doi: 10.1203/PDR.0b013e3181eb5d68.
Dominant mutations in coding regions of the surfactant protein-C gene, SFTPC, cause respiratory distress syndrome (RDS) in infants. However, the contribution of variants in noncoding regions of SFTPC to pulmonary phenotypes is unknown. By using a case-control group of infants > or =34 weeks gestation (n = 538), we used complete resequencing of SFTPC and its promoter, genotyping, and logistic regression to identify 80 single nucleotide polymorphisms (SNPs). Three promoter SNPs were statistically associated with neonatal RDS among European descent infants. To assess the transcriptional effects of these three promoter SNPs, we selectively mutated the SFTPC promoter and performed transient transfection using MLE-15 cells and a firefly luciferase reporter vector. Each promoter SNP decreased SFTPC transcription. The combination of two variants in high linkage dysequilibrium also decreased SFTPC transcription. In silico evaluation of transcription factor binding demonstrated that the rare allele at g.-1167 disrupts a SOX (SRY-related high mobility group box) consensus motif and introduces a GATA-1 site, at g.-2385 removes a MZF-1 (myeloid zinc finger) binding site, and at g.-1647 removes a potential methylation site. This combined statistical, in vitro, and in silico approach suggests that reduced SFTPC transcription contributes to the genetic risk for neonatal RDS in developmentally susceptible infants.
编码区域的显性突变表面活性蛋白-C 基因,SFTPC,导致呼吸窘迫综合征(RDS)在婴儿。然而,变异在非编码区域的 SFTPC 对肺表型是未知的。通过使用病例对照组的婴儿>或=34 周妊娠(n = 538),我们使用完整的测序 SFTPC 和其启动子,基因分型和逻辑回归来确定 80 个单核苷酸多态性(SNPs)。三个启动子 SNP 统计学上与新生儿 RDS 相关的欧洲血统的婴儿。为了评估这三个启动子 SNP 的转录效应,我们选择性地突变 SFTPC 启动子,并进行瞬时转染使用 MLE-15 细胞和萤火虫荧光素酶报告载体。每个启动子 SNP 降低 SFTPC 转录。两种变体在高连锁不平衡的组合也降低了 SFTPC 转录。转录因子结合的计算评估表明,在 g.-1167 处的罕见等位基因破坏了 SOX(SRY 相关的高迁移率组盒)的一个保守基序,并引入了一个 GATA-1 位点,在 g.-2385 处去除了一个 MZF-1(髓系锌指)结合位点,而在 g.-1647 处去除了一个潜在的甲基化位点。这种综合的统计、体外和计算机模拟方法表明,SFTPC 转录的降低导致新生儿 RDS 的遗传风险在发育易感婴儿。