MiR-548aq-3p is a novel target of Far infrared radiation which predicts coronary artery disease endothelial colony forming cell responsiveness

Wei-Che Tsai,Wei-Hui Chiang,Cheng-Chung Cheng,Shu-Meng Cheng,Po-Hsun Huang,Mel Campbell,Chi-Hung Lin,Pei-Ching Chang,Chun-Hsien Wu,Ming-Wei Lin,Yue-Cheng Li

doi:10.1038/s41598-020-63311-1

Abstract

Non-invasive far infrared radiation (FIR) has been observed to improve the health of patients with coronary artery disease (CAD). Endothelial colony forming cells (ECFCs) contribute to vascular repair and CAD. The goal of this study was to uncover the role of FIR in ECFCs function and to reveal potential biomarkers for indication of FIR therapy in CAD patients. FIR significantly enhanced in vitro migration (transwell assay) and tube formation (tube length) capacities in a subpopulation of CAD ECFCs. Clinical parameters associated with the responsiveness of ECFCs to FIR include smoking and gender. ECFCs from CAD patients that smoke did not respond to FIR in most cases. In contrast, ECFCs from females showed a higher responsiveness to FIR than ECFCs from males. To decipher the molecular mechanisms by which FIR modulates ECFCs functions, regardless of sex, RNA sequencing analysis was performed in both genders of FIR-responsive and FIR-non/unresponsive ECFCs. Gene Ontology (GO) analysis of FIR up-regulated genes indicated that the pathways enriched in FIR-responsive ECFCs were involved in cell viability, angiogenesis and transcription. Small RNA sequencing illustrated 18 and 14 miRNAs that are up- and down-regulated, respectively, in FIR-responsive CAD ECFCs in both genders. Among the top 5 up- and down-regulated miRNAs, down-regulation of miR-548aq-3p in CAD ECFCs after FIR treatment was observed in FIR-responsive CAD ECFCs by RT-qPCR. Down-regulation of miR-548aq-3p was correlated with the tube formation activity of CAD ECFCs enhanced by FIR. After establishment of the down-regulation of miR-548aq-3p by FIR in CAD ECFCs, we demonstrated through overexpression and knockdown experiments that miR-548aq-3p contributes to the inhibition of the tube formation of ECFCs. This study suggests the down-regulation of miR-548aq-3p by FIR may contribute to the improvement of ECFCs function, and represents a novel biomarker for therapeutic usage of FIR in CAD patients.

Highlights

Non-invasive far infrared radiation (FIR) has been observed to improve the health of patients with coronary artery disease (CAD)
By using 46 Coronary artery disease (CAD) Endothelial colony forming cells (ECFCs) isolated from CAD patients, the effect of Far infrared radiation (FIR) on the functionality of ECFCs was evaluated
Bioinformatics analysis of transcriptome data showed that FIR may modulate the angiogenesis function of CAD ECFCs. smRNA-seq revealed down-regulation of miR-548aq-3p may represent a unique mechanism to improve the tube formation ability of ECFCs following FIR treatment

Summary

Introduction

Non-invasive far infrared radiation (FIR) has been observed to improve the health of patients with coronary artery disease (CAD). After establishment of the down-regulation of miR-548aq-3p by FIR in CAD ECFCs, we demonstrated through overexpression and knockdown experiments that miR-548aq-3p contributes to the inhibition of the tube formation of ECFCs. This study suggests the down-regulation of miR-548aq-3p by FIR may contribute to the improvement of ECFCs function, and represents a novel biomarker for therapeutic usage of FIR in CAD patients. Small RNA sequencing (smRNA-seq) followed by RT-qPCR identified miR-548aq-3p as a FIR suppressed molecule that correlated with the FIR responsiveness of CAD ECFCs. Through experiments of functional validation, we further demonstrated that overexpression of miR-548aq-3p inhibited the tube formation of healthy ECFCs. In contrast, knockdown of miR-548aq-3p enhanced the tube formation of CAD ECFCs, suggesting this miRNA may contribute to FIR-mediated improvement of ECFCs function

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