Shigematsu Megumi, Kawamura Takuya, Deshpande Deepak A, Kirino Yohei
Computational Medicine Center, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Center for Translational Medicine, Jane and Leonard Korman Respiratory Institute, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Mol Ther Nucleic Acids. 2024 Jul 19;35(3):102285. doi: 10.1016/j.omtn.2024.102285. eCollection 2024 Sep 10.
Chronic obstructive pulmonary disease (COPD) is the most prevalent lung disease, and macrophages play a central role in the inflammatory response in COPD. We here report a comprehensive characterization of circulating short non-coding RNAs (sncRNAs) in plasma from patients with COPD. While circulating sncRNAs are increasingly recognized for their regulatory roles and biomarker potential in various diseases, the conventional RNA sequencing (RNA-seq) method cannot fully capture these circulating sncRNAs due to their heterogeneous terminal structures. By pre-treating the plasma RNAs with T4 polynucleotide kinase, which converts all RNAs to those with RNA-seq susceptible ends (5'-phosphate and 3'-hydroxyl), we comprehensively sequenced a wide variety of non-microRNA sncRNAs, such as 5'-tRNA halves containing a 2',3'-cyclic phosphate. We discovered a remarkable accumulation of the 5'-half derived from tRNA in plasma from COPD patients, whereas the 5'-tRNA half is predominant in healthy donors. Further, the 5'-tRNA half activates human macrophages via Toll-like receptor 7 and induces cytokine production. Additionally, we identified circulating rRNA-derived fragments that were upregulated in COPD patients and demonstrated their ability to induce cytokine production in macrophages. Our findings provide evidence of circulating, immune-active sncRNAs in patients with COPD, suggesting that they serve as inflammatory mediators in the pathogenesis of COPD.
慢性阻塞性肺疾病（COPD）是最常见的肺部疾病，巨噬细胞在COPD的炎症反应中起核心作用。我们在此报告了对COPD患者血浆中循环短链非编码RNA（sncRNAs）的全面表征。虽然循环sncRNAs因其在各种疾病中的调节作用和生物标志物潜力而越来越受到认可，但传统的RNA测序（RNA-seq）方法由于其异质的末端结构而无法完全捕获这些循环sncRNAs。通过用T4多核苷酸激酶预处理血浆RNA，将所有RNA转化为具有RNA-seq易感性末端（5'-磷酸和3'-羟基）的RNA，我们全面测序了多种非微小RNA的sncRNAs，例如含有2',3'-环磷酸的5'-tRNA半体。我们发现COPD患者血浆中来自tRNA的5'-半体显著积累，而在健康供体中5'-tRNA半体占主导。此外，5'-tRNA半体通过Toll样受体7激活人巨噬细胞并诱导细胞因子产生。此外，我们鉴定出在COPD患者中上调的循环rRNA衍生片段，并证明了它们在巨噬细胞中诱导细胞因子产生的能力。我们的研究结果提供了COPD患者中循环免疫活性sncRNAs的证据，表明它们在COPD发病机制中作为炎症介质起作用。
