Division of Respiratory Medicine, National Institute for Health Research, Nottingham Biomedical Research Centre, University of Nottingham, Nottingham, UK.
Respir Res. 2019 Aug 1;20(1):172. doi: 10.1186/s12931-019-1146-3.
Genome wide association (GWA) studies have reproducibly identified signals on chromosome 4q24 associated with lung function and COPD. GSTCD (Glutathione S-transferase C-terminal domain containing) represents a candidate causal gene in this locus, however little is currently known about the function of this protein. We set out to further our understanding of the role of GSTCD in cell functions and homeostasis using multiple molecular and cellular approaches in airway relevant cells. Recombinant expression of human GSTCD in conjunction with a GST activity assay did not identify any enzymatic activity for two GSTCD isoforms questioning the assignment of this protein to this family of enzymes. Protein structure analyses identified a potential methyltransferase domain contained within GSTCD, with these enzymes linked to cell viability and apoptosis. Targeted knockdown (siRNA) of GSTCD in bronchial epithelial cells identified a role for GSTCD in cell viability as proliferation rates were not altered. To provide greater insight we completed transcriptomic analyses on cells with GSTCD expression knocked down and identified several differentially expressed genes including those implicated in airway biology; fibrosis e.g. TGFBR1 and inflammation e.g. IL6R. Pathway based transcriptomic analyses identified an over-representation of genes related to adipogenesis which may suggest additional functions for GSTCD. These findings identify potential additional functions for GSTCD in the context of airway biology beyond the hypothesised GST activity and warrant further investigation.
全基因组关联 (GWA) 研究已经反复证实,4q24 染色体上的信号与肺功能和 COPD 相关。GSTCD(谷胱甘肽 S-转移酶 C 端结构域包含)是该基因座中候选因果基因,但目前对该蛋白的功能知之甚少。我们使用多种与气道相关的细胞中的分子和细胞方法,着手进一步了解 GSTCD 在细胞功能和动态平衡中的作用。人类 GSTCD 的重组表达与 GST 活性测定相结合,并未鉴定出两种 GSTCD 同工型的任何酶活性,这对将该蛋白分配到该酶家族提出了质疑。蛋白结构分析鉴定出 GSTCD 中包含一个潜在的甲基转移酶结构域,这些酶与细胞活力和细胞凋亡有关。在支气管上皮细胞中靶向敲低 (siRNA) GSTCD 发现 GSTCD 在细胞活力中发挥作用,因为增殖率没有改变。为了提供更深入的了解,我们对 GSTCD 表达敲低的细胞完成了转录组分析,鉴定出了几个差异表达的基因,包括与气道生物学相关的基因;纤维化,例如 TGFBR1 和炎症,例如 IL6R。基于途径的转录组分析鉴定出与脂肪生成相关的基因过度表达,这可能表明 GSTCD 具有其他功能。这些发现确定了 GSTCD 在气道生物学背景下除了假设的 GST 活性之外的潜在额外功能,并值得进一步研究。