Exosome-Mediated Crosstalk between Keratinocytes and Macrophages in Cutaneous Wound Healing.
Exosome-Mediated Crosstalk between Keratinocytes and Macrophages in Cutaneous Wound Healing.
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DOI：
10.1021/acsnano.0c03064
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发表时间：
2020-10-27
期刊：
影响因子：
17.1
通讯作者：
Ghatak S
中科院分区：
文献类型：
--
作者：
Zhou X;Brown BA;Siegel AP;El Masry MS;Zeng X;Song W;Das A;Khandelwal P;Clark A;Singh K;Guda PR;Gorain M;Timsina L;Xuan Y;Jacobson SC;Novotny MV;Roy S;Agarwal M;Lee RJ;Sen CK;Clemmer DE;Ghatak S
涉及外体携带的货物(如miRNA)的双向细胞-细胞通讯已成为伤口愈合的关键机制。与其他脱落小泡不同，Exosome通过异质核糖核蛋白A2B1(HnRNPA2B1)的SUMO化选择性地包装miRNA。在这项工作中，我们阐明了外切体在损伤后角质形成细胞-巨噬细胞串扰中的意义。用组织纳米转染法将角质形成细胞来源的外切体标记为绿色荧光蛋白报告基因(Exoκ-GFP)，并通过磁珠亲和选择从小鼠背部皮肤和创缘组织中分离得到。并对外源κ-绿色荧光蛋白的表面N-糖链进行了表征。与皮肤外体不同的是，伤口边缘的Exoκ-GFP显示了特有的N-糖离子和丰富的低碱基对核糖核酸，并选择性地被肉芽组织中的伤口巨噬细胞(ωmϕ)吞噬。在体外，将伤口边缘外源κ-绿色荧光蛋白加入到致炎的ωmϕ中，导致转化为前分解表型。为了在体内选择性地抑制外源κ-绿色荧光蛋白中miRNA的包装，设计了pH响应型角质形成细胞靶向siRNA-hnRNPA2B1功能化脂质纳米粒(κ)，包封率为94.3%。将TLNPκ/si-hnRNPA2B1应用于小鼠背部创缘，与TLNPκ/si对照组相比，可显著抑制表皮hnRNPA2B1的表达达80%。TLNPκ/si-hnRNPA2B1治疗组伤后第10天肉芽组织中炎性标志物ωmϕ的表达明显高于TLNPκ/si对照组，但创面愈合和再上皮化无明显差异。此外，TLNPκ/si-hnRNPA2B1治疗的小鼠表现出屏障功能受损，上皮连接蛋白表达减少，这证明了炎症未解决会导致皮肤渗漏的观点。这项工作提供了洞察力，其中外源性κ-绿色荧光蛋白被认为是调节巨噬细胞运输和损伤后上皮屏障属性的主要贡献者。
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 (Exoκ-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 Exoκ-GFP were also characterized. Unlike skin exosome, wound-edge Exoκ-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 Exoκ-GFP to proinflammatory ωmϕ resulted in conversion to a proresolution phenotype. To selectively inhibit miRNA packaging within Exoκ-GFP in vivo, pH-responsive keratinocyte-targeted siRNA-hnRNPA2B1 functionalized lipid nanoparticles (TLNPκ) were designed with 94.3% encapsulation efficiency. Application of TLNPκ/si-hnRNPA2B1 to murine dorsal wound-edge significantly inhibited expression of hnRNPA2B1 by 80% in epidermis compared to TLNPκ/si-control group. Although no significant difference in wound closure or re-epithelialization was observed, TLNPκ/si-hnRNPA2B1 treated group showed significant increase in ωmϕ displaying proinflammatory markers in the granulation tissue at day 10 post-wounding compared to TLNPκ/si-control group. Furthermore, TLNPκ/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 Exoκ-GFP are recognized as a major contributor that regulates macrophage trafficking and epithelial barrier properties post-injury.
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影响因子：
9
作者：
Barki KG;Das A;Dixith S;Ghatak PD;Mathew-Steiner S;Schwab E;Khanna S;Wozniak DJ;Roy S;Sen CK
通讯作者：
Sen CK
影响因子：
7.3
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通讯作者：
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影响因子：
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作者：
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影响因子：
1.2
作者：
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通讯作者：
Sanchez-Puig, Nuria
影响因子：
5.8
作者：
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通讯作者：
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