Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Feb 26, 2020; 12(2): 152-167
Published online Feb 26, 2020. doi: 10.4252/wjsc.v12.i2.152
C-C chemokine receptor type 2-overexpressing exosomes alleviated experimental post-stroke cognitive impairment by enhancing microglia/macrophage M2 polarization
Huai-Chun Yang, Min Zhang, Rui Wu, Hai-Qing Zheng, Li-Ying Zhang, Jing Luo, Li-Li Li, Xi-Quan Hu
Huai-Chun Yang, Rui Wu, Hai-Qing Zheng, Li-Ying Zhang, Jing Luo, Li-Li Li, Xi-Quan Hu, Department of Rehabilitation Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China
Min Zhang, Department of Andrology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China
Author contributions: Hu XH, Yang HC, and Zhang M conceived and designed the experiments. Yang HC and Zhang M performed the experiments. Yang HC, Zhang M and Wu R acquired, analyzed and interpreted the data. Hu XH, Yang HC, and Zhang M wrote the manuscript. All authors have read and revised the final manuscript and approved it for publication.
Supported by the National Natural Science Foundation of China, No. 81871847 and No. 81672261.
Institutional review board statement: The study was reviewed and approved by the Institutional Animal Ethics Committee of Life Sciences School, Sun Yat-sen University.
Institutional animal care and use committee statement: Animal studies were reviewed and approved by the Institutional Animal Ethics Committee of Life Sciences School, Sun Yat-sen University.
Conflict-of-interest statement: The authors declare no conflicts of interest.
Data sharing statement: The data used to support the findings of this study are available from the corresponding author upon request.
ARRIVE guidelines statement: The manuscript has been prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Xi-Quan Hu, MD, PhD, Chief Physician, Professor, Department of Rehabilitation Medicine, the Third Affiliated Hospital, Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou 510000, Guangdong Province, China. sysu_hu@163.com
Received: October 22, 2019
Peer-review started: October 22, 2019
First decision: November 18, 2019
Revised: December 27, 2019
Accepted: January 19, 2020
Article in press: January 19, 2020
Published online: February 26, 2020
Processing time: 128 Days and 3.2 Hours
Abstract
BACKGROUND

Human-derived mesenchymal stromal cells have been shown to improve cognitive function following experimental stroke. The activity of exosomes has been verified to be comparable to the therapeutic effects of mesenchymal stromal cells. However, the effects of exosomes derived from human umbilical cord mesenchymal stem cells (HUC-MSCs) (ExoCtrl) on post-stroke cognitive impairment (PSCI) have rarely been reported. Moreover, whether exosomes derived from C-C chemokine receptor type 2 (CCR2)-overexpressing HUC-MSCs (ExoCCR2) can enhance the therapeutic effects on PSCI and the possible underlying mechanisms have not been studied.

AIM

To investigate the effects of ExoCtrl on PSCI and whether ExoCCR2 can enhance therapeutic effects on PSCI.

METHODS

Transmission electron microscopy, qNano® particles analyzer, and Western blotting were employed to determine the morphology and CCR2 expression of ExoCtrl or ExoCCR2. ELISA was used to study the binding capacity of exosomes to CC chemokine ligand 2 (CCL2) in vivo. After the intravenous injection of ExoCtrl or ExoCCR2 into experimental rats, the effect of ExoCtrl and ExoCCR2 on PSCI was assessed by Morris water maze. Remyelination and oligodendrogenesis were analyzed by Western blotting and immunofluorescence microscopy. QRT-PCR and immunofluorescence microscopy were conducted to compare the microglia/macrophage polarization. The infiltration and activation of hematogenous macrophages were analyzed by Western blotting and transwell migration analysis.

RESULTS

CCR2-overexpressing HUC-MSCs loaded the CCR2 receptor into their exosomes. The morphology and diameter distribution between ExoCtrl and ExoCCR2 showed no significant difference. ExoCCR2 bound significantly to CCL2 but ExoCtrl showed little CCL2 binding. Although both ExoCCR2 and ExoCtrl showed beneficial effects on PSCI, oligodendrogenesis, remyelination, and microglia/macrophage polarization, ExoCCR2 exhibited a significantly superior beneficial effect. We also found that ExoCCR2 could suppress the CCL2-induced macrophage migration and activation in vivo and in vitro, compared with ExoCtrl treated group.

CONCLUSION

CCR2 over-expression enhanced the therapeutic effects of exosomes on the experimental PSCI by promoting M2 microglia/macrophage polarization, enhancing oligodendrogenesis and remyelination. These therapeutic effects are likely through suppressing the CCL2-induced hematogenous macrophage migration and activation.

Keywords: Cognitive impairment; Stroke; Exosomes; C-C chemokine receptor type 2; Microglia/macrophage polarization; Remyelination

Core tip: Exosomes have been reported to possess the therapeutic benefit comparable to the therapeutic effects of mesenchymal stromal cells. However, the effects of exosomes derived from human umbilical cord mesenchymal stem cells (ExoCtrl) on post-stroke cognitive impairment (PSCI) have rarely been reported. Moreover, whether exosomes derived from C-C chemokine receptor type 2 (CCR2)-overexpressing human umbilical cord mesenchymal stem cells (ExoCCR2) have better therapeutic effects on PSCI and the possible mechanisms underlying these effects remained unclear. This study provides new insights into the use of genetically modified exosomes for PSCI treatment, offering new ideas for the clinical application of exosome-based therapies for PSCI.