Defining the activity of pro-reparative extracellular vesicles in wound healing based on miRNA payloads and cell type-specific lineage mapping

Dong Jun Park; Wooil Choi; Sakeef Sayeed; Robert A Dorschner; Joseph Rainaldi; Kayla Ho; Jenny Kezios; John P Nolan; Prashant Mali; Todd Costantini; Brian P Eliceiri

doi:10.1016/j.ymthe.2024.02.019

Defining the activity of pro-reparative extracellular vesicles in wound healing based on miRNA payloads and cell type-specific lineage mapping

Mol Ther. 2024 Feb 19:S1525-0016(24)00088-1. doi: 10.1016/j.ymthe.2024.02.019. Online ahead of print.

Authors

Affiliations

¹ Department of Surgery, University of California San Diego, La Jolla, CA 92093, USA.
² Department of Dermatology, University of California San Diego, La Jolla, CA 92093, USA.
³ Department of Bioengineering, University of California San Diego, La Jolla, CA 92093, USA.
⁴ Scintillon Institute, San Diego, CA 92121, USA.
⁵ Department of Surgery, University of California San Diego, La Jolla, CA 92093, USA; Department of Dermatology, University of California San Diego, La Jolla, CA 92093, USA. Electronic address: beliceiri@health.ucsd.edu.

PMID: 38379282
DOI: 10.1016/j.ymthe.2024.02.019

Abstract

Small extracellular vesicles (EVs) are released by cells and deliver biologically active payloads to coordinate the response of multiple cell types in cutaneous wound healing. Here we used a cutaneous injury model as a donor of pro-reparative EVs to treat recipient diabetic obese mice, a model of impaired wound healing. We established a functional screen for microRNAs (miRNAs) that increased the pro-reparative activity of EVs and identified a down-regulation of miR-425-5p in EVs in vivo and in vitro associated with the regulation of adiponectin. We tested a cell type-specific reporter of a tetraspanin CD9 fusion with GFP to lineage map the release of EVs from macrophages in the wound bed, based on the expression of miR-425-5p in macrophage-derived EVs and the abundance of macrophages in EV donor sites. Analysis of different promoters demonstrated that EV release under the control of a macrophage-specific promoter was most abundant and that these EVs were internalized by dermal fibroblasts. These findings suggested that pro-reparative EVs deliver miRNAs, such as miR-425-5p, that stimulate the expression of adiponectin that has insulin-sensitizing properties. We propose that EVs promote intercellular signaling between cell layers in the skin to resolve inflammation, induce proliferation of basal keratinocytes, and accelerate wound closure.

Keywords: adiponectin; diabetic wound closure; extracellular vesicles; macrophage; miR-425-5p; single-cell RNA sequence.