LCK rs10914542-G 等位基因通过 T 细胞低反应性与儿童 1 型糖尿病相关联。

LCK rs10914542-G allele associates with type 1 diabetes in children via T cell hyporesponsiveness.

机构信息

Prenatal Screening and Diagnosis Center, Nantong Municipal Maternal and Child Health Hospital, 226010, Nantong, China.

出版信息

Pediatr Res. 2019 Sep;86(3):311-315. doi: 10.1038/s41390-019-0436-2. Epub 2019 May 21.

DOI:10.1038/s41390-019-0436-2

PMID:31112992

Abstract

BACKGROUND

Abnormal lymphocyte-specific protein tyrosine kinase (LCK)-related T cell hyporesponsiveness was discovered in type 1 diabetes (T1D). This study aims to investigate the potential associations between LCK single-nucleotide polymorphisms (SNPs) and the susceptibility of T1D.

METHODS

DNAs were extracted from blood samples of 589 T1D patients and 596 healthy controls to genotype seven SNPs of the LCK gene using PCR and Sanger sequencing. Associations of these SNPs with the susceptibility of T1D were determined by χ test. LCKs were knocked out in peripheral blood mononuclear cells (PBMCs) using CRISPR-Cas9 to investigate the role of LCK SNP in T-lymphocyte activation in T1D.

RESULTS

SNP rs10914542 but not the other six SNPs of the LCK gene was significantly associated with (C vs. G, odds ratio (OR) = 0.581, 95% confidence interval (CI) = 0.470-0.718, P value = 4.13E - 7) the susceptibility of T1D. Peripheral T-lymphocyte activation in response to T cell receptor (TCR)/CD3 stimulation is significantly lower in the rs10914542-G-allele group than in the C-allele group. In vitro experiments revealed that rs10914542 G allele impaired the TCR/CD3-mediated T-cell activation in PBMCs.

CONCLUSIONS

This study reveals that the G allele of SNP rs10914542 of LCK impairs the TCR/CD3-mediated T-cell activation and increases the risk of T1D.

摘要

背景

1 型糖尿病（T1D）患者中发现了异常淋巴细胞特异性蛋白酪氨酸激酶（LCK）相关 T 细胞反应低下。本研究旨在探讨 LCK 单核苷酸多态性（SNP）与 T1D 易感性之间的潜在关联。

方法

从 589 例 T1D 患者和 596 例健康对照者的血液样本中提取 DNA，采用 PCR 和 Sanger 测序方法对 LCK 基因的 7 个 SNP 进行基因分型。采用 χ 检验确定这些 SNP 与 T1D 易感性的关系。使用 CRISPR-Cas9 敲除外周血单个核细胞（PBMC）中的 LCK，以研究 LCK SNP 在 T1D 中 T 淋巴细胞活化中的作用。

结果

LCK 基因的 SNP rs10914542 与 T1D 的易感性显著相关（C 与 G，比值比（OR）=0.581，95%置信区间（CI）=0.470-0.718，P 值=4.13E-7），但其他六个 LCK 基因的 SNP 则无显著相关性。T 细胞受体（TCR）/CD3 刺激后外周 T 淋巴细胞的活化在 rs10914542-G 等位基因组明显低于 C 等位基因组。体外实验表明，rs10914542 G 等位基因损害了 TCR/CD3 介导的 PBMC 中的 T 细胞活化。