Importance of microRNAs by mRNA-microRNA integration analysis in acute ischemic stroke patients

Hideaki Kanki; Hisatake Matsumoto; Yuki Togami; Daisuke Okuzaki; Hiroshi Ogura; Tsutomu Sasaki; Hideki Mochizuki

doi:10.1016/j.jstrokecerebrovasdis.2023.107277

Importance of microRNAs by mRNA-microRNA integration analysis in acute ischemic stroke patients

J Stroke Cerebrovasc Dis. 2023 Sep;32(9):107277. doi: 10.1016/j.jstrokecerebrovasdis.2023.107277. Epub 2023 Aug 8.

Authors

Hideaki Kanki¹, Hisatake Matsumoto², Yuki Togami², Daisuke Okuzaki³, Hiroshi Ogura², Tsutomu Sasaki⁴, Hideki Mochizuki¹

Affiliations

¹ Department of Neurology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
² Department of Traumatology and Acute Critical Medicine, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan.
³ Genome Information Research Center, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
⁴ Department of Neurology, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan; Department of Neurotherapeutics, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan. Electronic address: sasaki@neurol.med.osaka-u.ac.jp.

PMID: 37562178
DOI: 10.1016/j.jstrokecerebrovasdis.2023.107277

Abstract

Objects: 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.

Material and methods: 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.

Results: 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.

Conclusions: 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.

Keywords: Ischemic stroke; MSP-RON signaling; RNA sequencing; mRNA; mRNA and miRNA integration analysis; microRNA.