Exosome-Mediated Crosstalk between Keratinocytes and Macrophages in Cutaneous Wound Healing

Xiaoju Zhou; Brooke A Brown; Amanda P Siegel; Mohamed S El Masry; Xuyao Zeng; Woran Song; Amitava Das; Puneet Khandelwal; Andrew Clark; Kanhaiya Singh; Poornachander R Guda; Mahadeo Gorain; Lava Timsina; Yi Xuan; Stephen C Jacobson; Milos V Novotny; Sashwati Roy; Mangilal Agarwal; Robert J Lee; Chandan K Sen; David E Clemmer; Subhadip Ghatak

doi:10.1021/acsnano.0c03064

Exosome-Mediated Crosstalk between Keratinocytes and Macrophages in Cutaneous Wound Healing

ACS Nano. 2020 Oct 27;14(10):12732-12748. doi: 10.1021/acsnano.0c03064. Epub 2020 Sep 25.

Authors

Xiaoju Zhou^{1

2}, Brooke A Brown³, Amanda P Siegel^{1

4}, Mohamed S El Masry^{1

5}, Xuyao Zeng³, Woran Song³, Amitava Das¹, Puneet Khandelwal¹, Andrew Clark¹, Kanhaiya Singh¹, Poornachander R Guda¹, Mahadeo Gorain¹, Lava Timsina^{1

6}, Yi Xuan¹, Stephen C Jacobson³, Milos V Novotny³, Sashwati Roy¹, Mangilal Agarwal⁴, Robert J Lee⁷, Chandan K Sen¹, David E Clemmer³, Subhadip Ghatak¹

Affiliations

¹ Indiana Center for Regenerative Medicine & Engineering, Department of Surgery, Indiana University School of Medicine, Indianapolis, Indiana 46202, United States.
² School of Pharmaceutical Sciences, Wuhan University, Wuhan, 430071, China.
³ Department of Chemistry, Indiana University, Bloomington, Indiana 47405, United States.
⁴ Integrated Nanosystems Development Institute, Indiana University-Purdue University, Indianapolis, Indiana 46202, United States.
⁵ Department of Plastic and Reconstructive Surgery, Zagazig University, Zagazig, 44519, Egypt.
⁶ Center for Outcomes Research, Department of Surgery, Indiana University School of Medicine, Indianapolis, Indiana 46202, United States.
⁷ Division of Pharmaceutics and Pharmacology, The Ohio State University, Columbus, Ohio 43210, United States.

Abstract

Bidirectional cell-cell communication involving exosome-borne cargo such as miRNA has emerged as a critical mechanism for wound healing. Unlike other shedding vesicles, exosomes selectively package miRNA by SUMOylation of heterogeneous nuclear ribonucleoproteinA2B1 (hnRNPA2B1). In this work, we elucidate the significance of exosome in keratinocyte-macrophage crosstalk following injury. Keratinocyte-derived exosomes were genetically labeled with GFP-reporter (Exo_κ-GFP) using tissue nanotransfection (TNT), and they were isolated from dorsal murine skin and wound-edge tissue by affinity selection using magnetic beads. Surface N-glycans of Exo_κ-GFP were also characterized. Unlike skin exosome, wound-edge Exo_κ-GFP demonstrated characteristic N-glycan ions with abundance of low-base-pair RNA and was selectively engulfed by wound macrophages (ωmϕ) in granulation tissue. In vitro addition of wound-edge Exo_κ-GFP to proinflammatory ωmϕ resulted in conversion to a proresolution phenotype. To selectively inhibit miRNA packaging within Exo_κ-GFPin vivo, pH-responsive keratinocyte-targeted siRNA-hnRNPA2B1 functionalized lipid nanoparticles (TLNP_κ) were designed with 94.3% encapsulation efficiency. Application of TLNP_{κ/si-hnRNPA2B1} to the murine dorsal wound-edge significantly inhibited expression of hnRNPA2B1 by 80% in epidermis compared to the TLNP_{κ/si-control} group. Although no significant difference in wound closure or re-epithelialization was observed, the TLNP_{κ/si-hnRNPA2B1} treated group showed a significant increase in ωmϕ displaying proinflammatory markers in the granulation tissue at day 10 post-wounding compared to the TLNP_{κ/si-control} group. Furthermore, TLNP_{κ/si-hnRNPA2B1} treated mice showed impaired barrier function with diminished expression of epithelial junctional proteins, lending credence to the notion that unresolved inflammation results in leaky skin. This work provides insight wherein Exo_κ-GFP is recognized as a major contributor that regulates macrophage trafficking and epithelial barrier properties postinjury.

Keywords: exosome; keratinocyte-targeted lipid nanoparticles; macrophage; tissue nanotransfection; wound healing.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Exosomes*
Keratinocytes
Macrophages
Mice
Skin
Wound Healing

Abstract

Publication types

MeSH terms

Grants and funding