Human umbilical cord mesenchymal stem cell-derived exosomes promote murine skin wound healing by neutrophil and macrophage modulations revealed by single-cell RNA sequencing

Yuanyuan Liu; Mingwang Zhang; Yong Liao; Hongbo Chen; Dandan Su; Yuandong Tao; Jiangbo Li; Kai Luo; Lihua Wu; Xingyue Zhang; Rongya Yang

doi:10.3389/fimmu.2023.1142088

Human umbilical cord mesenchymal stem cell-derived exosomes promote murine skin wound healing by neutrophil and macrophage modulations revealed by single-cell RNA sequencing

Front Immunol. 2023 Mar 6:14:1142088. doi: 10.3389/fimmu.2023.1142088. eCollection 2023.

Authors

Yuanyuan Liu^{1

2}, Mingwang Zhang³, Yong Liao², Hongbo Chen⁴, Dandan Su⁴, Yuandong Tao⁵, Jiangbo Li⁶, Kai Luo⁷, Lihua Wu⁷, Xingyue Zhang², Rongya Yang²

Affiliations

¹ Medical School of Chinese People's Liberation Army, Beijing, China.
² Department of Dermatology, the Seventh Medical Center of Chinese People's Liberation Army (PLA) General Hospital, Beijing, China.
³ Department of Dermatology, Southwest Hospital, Army Medical University, Chongqing, China.
⁴ School of Pharmaceutical Sciences, Sun Yat-sen University, Shenzhen, China.
⁵ Department of Pediatric Urology, the Seventh Medical Center of Chinese People's Liberation Army (PLA) General Hospital, Beijing, China.
⁶ Bioinformatics Center of Academy of Military Medical Sciences, Beijing, China.
⁷ Biomedical Treatment Center, the Seventh Medical Center of Chinese People's Liberation Army (PLA) General Hospital, Beijing, China.

Abstract

Introduction: Full-thickness skin wound healing remains a serious undertaking for patients. While stem cell-derived exosomes have been proposed as a potential therapeutic approach, the underlying mechanism of action has yet to be fully elucidated. The current study aimed to investigate the impact of exosomes derived from human umbilical cord mesenchymal stem cells (hucMSC-Exosomes) on the single-cell transcriptome of neutrophils and macrophages in the context of wound healing.

Methods: Utilizing single-cell RNA sequencing, the transcriptomic diversity of neutrophils and macrophages was analyzed in order to predict the cellular fate of these immune cells under the influence of hucMSC-Exosomes and to identify alterations of ligand-receptor interactions that may influence the wound microenvironment. The validity of the findings obtained from this analysis was subsequently corroborated by immunofluorescence, ELISA, and qRT-PCR. Neutrophil origins were characterized based on RNA velocity profiles.

Results: The expression of RETNLG and SLC2A3 was associated with migrating neutrophils, while BCL2A1B was linked to proliferating neutrophils. The hucMSC-Exosomes group exhibited significantly higher levels of M1 macrophages (215 vs 76, p < 0.00001), M2 macrophages (1231 vs 670, p < 0.00001), and neutrophils (930 vs 157, p < 0.00001) when compared to control group. Additionally, it was observed that hucMSC-Exosomes elicit alterations in the differentiation trajectories of macrophages towards more anti-inflammatory phenotypes, concomitant with changes in ligand-receptor interactions, thereby facilitating healing.

Discussion: This study has revealed the transcriptomic heterogeneity of neutrophils and macrophages in the context of skin wound repair following hucMSC-Exosomes interventions, providing a deeper understanding of cellular responses to hucMSC-Exosomes, a rising target of wound healing intervention.

Keywords: cellular heterogeneity; exosomes; macrophages; neutrophils; single-cell RNA sequencing; wound healing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Exosomes* / genetics
Exosomes* / metabolism
Humans
Ligands
Macrophages / metabolism
Mesenchymal Stem Cells* / metabolism
Mice
Neutrophils
Sequence Analysis, RNA
Umbilical Cord
Wound Healing / genetics

Substances

Ligands

Grants and funding

This work was supported by the National Science Foundation of China (81571972) and the National Science Foundation of Chongqing, China (No. cstc2021jcyj-msxmX0140).