Department of Oral Science, Graduate School of Medicine, Chiba University, Chiba, Japan.
Department of Dentistry and Oral-Maxillofacial Surgery, Chiba University Hospital, Chiba, Japan.
Biochem Biophys Res Commun. 2022 Jan 8;587:146-152. doi: 10.1016/j.bbrc.2021.11.100. Epub 2021 Nov 30.
Lysyl hydroxylase 2 (LH2) regulates intermolecular cross-linking of collagen molecules. Accumulation of LH2-modified collagen, which is highly stable and resistant to collagenase cleavage, is one cause of fibrosis. We previously demonstrated that conventional LH2 knockout mice showed embryonic lethality. Here we established LH2 conditional knockout mice using a tamoxifen-inducible Cre system. Morphological analysis of LH2-deficient fibroblasts by microscopy showed a dramatic increase in the number of filopodia, the finger-like cell surface projections that enable cell movement. The tips and leading edges of these filopodia exhibited up-regulated expression of Myosin-X (Myo10), a regulator of filopodial integrity. Wound healing assays demonstrated that migration of LH2-deficient cells was significantly faster than that of control cells. Gene expression profiling data also supported this phenotype. Together these findings indicate that LH2 deficiency may prevent fibrosis through decreased accumulation of LH2-cross-linked collagen, and that fibroblasts with faster migration contribute to enhanced wound healing activity. In conclusion, our cellular models provide evidence that LH2 deficiency plays a critical role in cell migration mediated through filopodia formation. Understanding the precise role of this phenotype in LH2-deficient cells may be helpful to define the pathogenesis of fibrosis. As such, detailed analyses of fibrosis and wound healing using LH2-deficient mouse models are needed.
赖氨酰羟化酶 2(LH2)调节胶原蛋白分子的分子间交联。LH2 修饰的胶原蛋白的积累是纤维化的一个原因,这种胶原蛋白高度稳定,不易被胶原酶切割。我们之前证明了常规的 LH2 敲除小鼠表现出胚胎致死性。在这里,我们使用他莫昔芬诱导的 Cre 系统建立了 LH2 条件性敲除小鼠。通过显微镜对 LH2 缺陷型成纤维细胞的形态分析表明,丝状伪足的数量显著增加,丝状伪足是一种使细胞运动的细胞表面突起。这些丝状伪足的尖端和前缘表现出肌球蛋白 X(Myo10)的上调表达,Myo10 是丝状伪足完整性的调节剂。伤口愈合试验表明,LH2 缺陷型细胞的迁移速度明显快于对照细胞。基因表达谱数据也支持这种表型。这些发现表明,LH2 缺乏可能通过减少 LH2 交联的胶原蛋白积累来预防纤维化,并且迁移速度更快的成纤维细胞有助于增强伤口愈合活性。总之,我们的细胞模型为 LH2 缺乏通过形成丝状伪足介导的细胞迁移中发挥关键作用提供了证据。了解这种表型在 LH2 缺陷型细胞中的精确作用可能有助于确定纤维化的发病机制。因此,需要使用 LH2 缺陷型小鼠模型对纤维化和伤口愈合进行详细分析。