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

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

World J Stem Cells. 2020 Feb 26;12(2):152-167. doi: 10.4252/wjsc.v12.i2.152.

Authors

Huai-Chun Yang¹, Min Zhang², Rui Wu¹, Hai-Qing Zheng¹, Li-Ying Zhang¹, Jing Luo¹, Li-Li Li¹, Xi-Quan Hu³

Affiliations

¹ Department of Rehabilitation Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China.
² Department of Andrology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China.
³ Department of Rehabilitation Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China. sysu_hu@163.com.

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) (Exo^Ctrl) 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 (Exo^CCR2) can enhance the therapeutic effects on PSCI and the possible underlying mechanisms have not been studied.

Aim: To investigate the effects of Exo^Ctrl on PSCI and whether Exo^CCR2 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 Exo^Ctrl or Exo^CCR2. ELISA was used to study the binding capacity of exosomes to CC chemokine ligand 2 (CCL2) in vivo. After the intravenous injection of Exo^Ctrl or Exo^CCR2 into experimental rats, the effect of Exo^Ctrl and Exo^CCR2 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 Exo^Ctrl and Exo^CCR2 showed no significant difference. Exo^CCR2 bound significantly to CCL2 but Exo^Ctrl showed little CCL2 binding. Although both Exo^CCR2 and Exo^Ctrl showed beneficial effects on PSCI, oligodendrogenesis, remyelination, and microglia/macrophage polarization, Exo^CCR2 exhibited a significantly superior beneficial effect. We also found that Exo^CCR2 could suppress the CCL2-induced macrophage migration and activation in vivo and in vitro, compared with Exo^Ctrl 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: C-C chemokine receptor type 2; Cognitive impairment; Exosomes; Microglia/macrophage polarization; Remyelination; Stroke.