Molecular Genetics Unit, Instituto de Investigación de Enfermedades Raras (IIER), Instituto de Salud Carlos III (ISCIII), CIBER de Enfermedades Raras (CIBERER), Madrid, Spain.
Respiratory Medicine Department, Coventry University Hospital, Coventry, UK.
Arch Bronconeumol. 2021 Jul;57(7):457-463. doi: 10.1016/j.arbr.2020.03.031.
Alpha-1 antitrypsin deficiency (AATD) is a genetic condition resulting in lung and liver disease with a great clinical variability. MicroRNAs have been identified as disease modifiers; therefore miRNA deregulation could play an important role in disease heterogeneity. Members of miR-320 family are involved in regulating of multiple processes including inflammation, and have potential specific binding sites in the 3'UTR region of SERPINA1 gene. In this study we explore the involvement of miR-320c, a member of this family, in this disease.
Firstly in vitro studies were carried out to demonstrate regulation of SERPINA1 gene by miR-320. Furthermore, the expression of miR-320c was analyzed in the blood of 98 individuals with different AAT serum levels by using quantitative PCR and expression was correlated to clinical parameters of the patients. Finally, HL60 cells were used to analyze induction of miR-320c in inflammatory conditions.
Overexpression of miR-320 members in human HepG2 cells led to inhibition of SERPINA1 expression. Analysis of miR-320c expression in patient's samples revealed significantly increased expression of miR-320c in individuals with pulmonary disease. Additionally, HL60 cells treated with the pro-inflammatory factor lipopolysaccharide (LPS) showed increase in miR-320c expression, suggesting that miR-320c responds to inflammation.
Our findings demonstrate that miR-320c inhibits SERPINA1 expression in a hepatic cell line and its levels in blood are associated with lung disease in a cohort of patients with different AAT serum levels. These results suggest that miR-320c can play a role in AAT regulation and could be a biomarker of inflammatory processes in pulmonary diseases.
α-1 抗胰蛋白酶缺乏症（AATD）是一种导致肺部和肝脏疾病的遗传病症，具有很大的临床变异性。 microRNAs 已被确定为疾病修饰因子；因此，miRNA 的失调可能在疾病异质性中发挥重要作用。miR-320 家族的成员参与调节多种过程，包括炎症，并且在 SERPINA1 基因的 3'UTR 区域具有潜在的特异性结合位点。在这项研究中，我们探讨了该家族成员 miR-320c 在该疾病中的作用。
首先进行了体外研究，以证明 miR-320 对 SERPINA1 基因的调节作用。此外，通过定量 PCR 分析了 98 名具有不同 AAT 血清水平的个体血液中的 miR-320c 表达，并将表达与患者的临床参数相关联。最后，使用 HL60 细胞分析炎症条件下 miR-320c 的诱导。
在人 HepG2 细胞中过表达 miR-320 成员导致 SERPINA1 表达受到抑制。在患者样本中分析 miR-320c 的表达显示，在患有肺部疾病的个体中，miR-320c 的表达显著增加。此外，用促炎因子脂多糖（LPS）处理的 HL60 细胞显示 miR-320c 表达增加，表明 miR-320c 对炎症有反应。
我们的研究结果表明，miR-320c 在肝系细胞中抑制 SERPINA1 的表达，其在血液中的水平与不同 AAT 血清水平患者的肺部疾病有关。这些结果表明，miR-320c 可在 AAT 调节中发挥作用，并且可能是肺部疾病中炎症过程的生物标志物。
