Kanki Hideaki, Matsumoto Hisatake, Togami Yuki, Okuzaki Daisuke, Ogura Hiroshi, Sasaki Tsutomu, Mochizuki Hideki
Department of Neurology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
Department of Traumatology and Acute Critical Medicine, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
J Stroke Cerebrovasc Dis. 2023 Sep;32(9):107277. doi: 10.1016/j.jstrokecerebrovasdis.2023.107277. Epub 2023 Aug 8.
The roles of mRNA and microRNA (miRNA) are well known in many diseases, including ischemic stroke; thus, integration analysis using mRNA and miRNA is important to elucidate pathogenesis. However, their contribution, especially that of miRNA-targeted mRNA, to the severity of acute ischemic stroke remains unclear. Therefore, we examined mRNA and miRNA integration analysis targeted for acute ischemic stroke to clarify the pathway related to acute stroke severity.
We performed Ingenuity Pathway Analysis (IPA) using RNA extracted from the whole blood of four healthy controls, six minor acute ischemic stroke patients (MS; National Institutes of Health Stroke Scale [NIHSS] < 8), and six severe acute ischemic stroke patients (SS; NIHSS ≥ 8) on admission. mRNA and miRNA were measured using RNA sequencing and RNA expression variation; canonical pathway analysis (CPA) and upstream regulator analyses were performed.
Acute ischemic stroke patients demonstrated different RNA expressions to healthy controls. Compared to MS patients, in the SS patients, 1222 mRNA, 96 miRNA, and 935 miRNA-targeted mRNA expressions were identified among differentially expressed RNA expressions (p<0.05, |log2 fold change| >1.1). CPA by IPA using mRNAs or miRNA-targeted mRNAs showed that macrophage-stimulating protein (MSP)-recepteur d'origine nantais (RON) signaling was mostly activated in SS patients compared to in MS patients. In addition, upstream regulator analysis in IPA showed that most mRNAs located upstream are miRNAs.
In severe acute stroke, integration of mRNA and microRNA analysis showed activated MSP-RON signaling in macrophages, and multiple miRNAs comprehensively controlled the overall pathophysiology of stroke.
信使核糖核酸（mRNA）和微小核糖核酸（miRNA）在包括缺血性中风在内的多种疾病中的作用已为人熟知；因此，利用mRNA和miRNA进行整合分析对于阐明发病机制很重要。然而，它们对急性缺血性中风严重程度的贡献，尤其是miRNA靶向mRNA的贡献仍不清楚。因此，我们对急性缺血性中风进行了mRNA和miRNA整合分析，以阐明与急性中风严重程度相关的途径。
我们对4名健康对照者、6名轻度急性缺血性中风患者（MS组；美国国立卫生研究院卒中量表[NIHSS]<8）和6名重度急性缺血性中风患者（SS组；NIHSS≥8）入院时采集的全血提取的RNA进行了 Ingenuity 通路分析（IPA）。使用RNA测序和RNA表达变异来测量mRNA和miRNA；进行了经典通路分析（CPA）和上游调节因子分析。
急性缺血性中风患者表现出与健康对照者不同的RNA表达。与MS组患者相比，在SS组患者中，在差异表达的RNA中鉴定出1222种mRNA、96种miRNA和935种miRNA靶向的mRNA表达（p<0.05，|log2倍数变化|>1.1）。使用mRNA或miRNA靶向的mRNA通过IPA进行的CPA显示，与MS组患者相比，巨噬细胞刺激蛋白（MSP）-源自南特的受体（RON）信号通路在SS组患者中大多被激活。此外，IPA中的上游调节因子分析表明，位于上游的大多数mRNA是miRNA。
在重度急性中风中，mRNA和微小核糖核酸分析的整合显示巨噬细胞中MSP-RON信号通路被激活，并且多种miRNA全面控制中风的整体病理生理学。
