Exosome-Mediated Crosstalk between Keratinocytes and Macrophages in Cutaneous Wound Healing
| Autor: | Lava Timsina, Puneet Khandelwal, Mohamed S. El Masry, Stephen C. Jacobson, Subhadip Ghatak, Amanda P. Siegel, Amitava Das, Mahadeo Gorain, David E. Clemmer, Xuyao Zeng, Sashwati Roy, Robert J. Lee, Brooke A. Brown, Xiaoju Zhou, Andrew G. Clark, Chandan K. Sen, Woran Song, Kanhaiya Singh, Yi Xuan, Mangilal Agarwal, Poornachander R. Guda, Milos V. Novotny |
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| Rok vydání: | 2020 |
| Předmět: |
Keratinocytes
General Physics and Astronomy 02 engineering and technology 010402 general chemistry Exosomes 01 natural sciences Exosome Article Mice microRNA Animals General Materials Science Skin Wound Healing Chemistry Vesicle Macrophages General Engineering 021001 nanoscience & nanotechnology Microvesicles 0104 chemical sciences Cell biology Crosstalk (biology) Cutaneous wound 0210 nano-technology Wound healing |
| Zdroj: | ACS Nano |
| ISSN: | 1936-086X |
| Popis: | 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 and 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 were 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-GFP) in 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 murine dorsal wound-edge significantly inhibited expression of hnRNPA2B1 by 80% in epidermis compared to TLNP(κ/si-control) group. Although no significant difference in wound closure or re-epithelialization was observed, TLNP(κ/si-hnRNPA2B1) treated group showed significant increase in ωmϕ displaying proinflammatory markers in the granulation tissue at day 10 post-wounding compared to 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) are recognized as a major contributor that regulates macrophage trafficking and epithelial barrier properties post-injury. |
| Databáze: | OpenAIRE |
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