Division of Life Sciences, College of Life Sciences and Biotechnology, Korea University, 145 Anam-dong, Sungbuk-ku, Seoul 136701, South Korea.
Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, 145 Anam-dong, Sungbuk-ku, Seoul 136701, South Korea.
Wound Repair Regen. 2019 Jul;27(4):345-359. doi: 10.1111/wrr.12713. Epub 2019 Apr 4.
Proliferative and migratory abilities of fibroblasts are essential for wound healing at the skin surface. Cytoplasmic linker-associated protein-2 (CLASP2) was originally found to interact with cytoplasmic linker protein (CLIP)-170. CLASP2 plays an important role in microtubule stabilization and the microtubule-stabilizing activity of CLASP2 depends on its interactions with end binding (EB)-1 and CLIP-170. Although the microtubule-stabilizing role of CLASP2 is well established, the effects of CLASP2 on the migration and proliferation of fibroblasts remain unclear in the context of wound healing. Therefore, we tested the utilization of CLASP2 as a directly applied protein drug to improve wound healing by promoting the migration of effector cells, including skin fibroblasts, to the site of repair or injury using an in vivo excisional wound mouse model and in vitro Hs27 skin fibroblast model. Epidermal growth factor, which is a recognized contributor to cell proliferation and migration, was used as positive control. In vitro and in vivo, CLASP2 treatment significantly enhanced cell migration and accelerated wound closure. Furthermore, in vivo, the CLASP2-treated animal group displayed enhanced epidermal repair and collagen deposition. Next, we studied the mechanism of CLASP2 for wound healing. Increasing the abundance of intracellular free CLASP2 in skin fibroblasts by supplying exogenous CLASP2 seemed to stabilize microtubules through an interaction between CLASP2 and CLIP-170, as well as EB1. Exogenous CLASP2 also showed direct binding with IQGAP1, increasing both cyclic adenosine monophosphate activity and phosphorylation of glycogen synthase kinase 3β, which in turn reinstated the binding between free CLASP2 and IQGAP1. In summary, exogenous CLASP2 increased Hs27 skin fibroblast migration by interacting with IQGAP1 and other cytoskeletal linker proteins, such as CLIP-170 and EB1. Our results strongly suggest that CLASP2 can be developed in wound healing drugs for skin repair and/or regenerating cosmetic products.
成纤维细胞的增殖和迁移能力对于皮肤表面的伤口愈合至关重要。细胞质连接蛋白-2（CLASP2）最初被发现与细胞质连接蛋白（CLIP）-170相互作用。CLASP2 在微管稳定中发挥重要作用，CLASP2 的微管稳定活性取决于其与末端结合蛋白（EB）-1 和 CLIP-170 的相互作用。尽管 CLASP2 的微管稳定作用已得到充分证实，但在伤口愈合的背景下，CLASP2 对成纤维细胞迁移和增殖的影响仍不清楚。因此，我们使用体内切除性伤口小鼠模型和体外 Hs27 皮肤成纤维细胞模型，测试了将 CLASP2 用作直接应用的蛋白质药物，通过促进包括皮肤成纤维细胞在内的效应细胞迁移到修复或损伤部位，来改善伤口愈合的可能性。表皮生长因子被用作细胞增殖和迁移的公认促进剂。在体外和体内，CLASP2 处理均显著增强了细胞迁移并加速了伤口闭合。此外，在体内，CLASP2 处理的动物组显示出增强的表皮修复和胶原蛋白沉积。接下来，我们研究了 CLASP2 促进伤口愈合的机制。通过提供外源性 CLASP2 增加皮肤成纤维细胞内游离 CLASP2 的丰度，似乎通过 CLASP2 与 CLIP-170 以及 EB1 的相互作用稳定了微管。外源性 CLASP2 还与 IQGAP1 直接结合，增加环磷酸腺苷活性和糖原合酶激酶 3β 的磷酸化，进而重新建立游离 CLASP2 与 IQGAP1 之间的结合。总之，外源性 CLASP2 通过与 IQGAP1 以及其他细胞骨架连接蛋白（如 CLIP-170 和 EB1）相互作用，增加了 Hs27 皮肤成纤维细胞的迁移。我们的结果强烈表明，CLASP2 可用于开发用于皮肤修复和/或再生美容产品的伤口愈合药物。
