Development of a new treatment strategy for wound healing using recombinant type VII collagen

使用重组 VII 型胶原蛋白开发伤口愈合新治疗策略

• 批准号: 15390336
• 负责人: SHIMIZU Hiroshi
• 金额: $ 9.47万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390336/
• 关键词: wound healing; epidermolvsis bullosa; dystrophic; severe; type VII collagen; protein therapy; gene transfer; skin ulcer

Type VII collagen, a non-fibrillar collagen, is a major component of anchoring fibril structures beneath the epidermal basement membrane, and is of critical importance for integrity of the cutaneous basement membrane zone. Mutations within the type VII collagen gene are associated with the dystrophic forms of epidermolysis bullosa (DEB), a group of mechanobullous diseases characterized by cutaneous fragility and tendency to form sub-basal lamina densa blisters. The patients with DEB showed delayd wound healing. The aim of this study was to clarify therapeutic effect of recombinant type VII collagen on wound healing. First, we purified a large amount of recombinant type VII collagen. We introduced the type VII collagen gene into HaCaT cells, NIH3T3 cells and 293 cells and performed selection of stable transformants by G418.293 cells expressed type VII collagen mostly among those cell lines. We could show low mobility of keratinocytes and fibroblasts from type VII collagen-gene knockout mouse and DEB patients, but addtion of recombinant type VII collagen rescued impaired mobility. Furthermore, application of recombinant type VII collagen to skin ulcer improved wound healing and we proposed type VII collagen as an anti-ulcer drug. Thus, we think that we can achieve establishment of a new strategy for treatment of wound using recombinant type VII collagen.

VII 型胶原蛋白是一种非纤维状胶原蛋白，是锚定表皮基底膜下方纤维结构的主要成分，对于皮肤基底膜区域的完整性至关重要。 VII 型胶原蛋白基因内的突变与营养不良型大疱性表皮松解症 (DEB) 相关，DEB 是一组以皮肤脆弱和形成基底致密层水疱倾向为特征的机械性大疱性疾病。 DEB 患者表现出伤口愈合延迟。本研究的目的是阐明重组 VII 型胶原蛋白对伤口愈合的治疗作用。首先，我们纯化了大量的重组VII型胶原蛋白。我们将VII型胶原蛋白基因导入HaCaT细胞、NIH3T3细胞和293细胞中，并通过G418进行稳定转化体的选择。在这些细胞系中，293细胞主要表达VII型胶原蛋白。我们发现 VII 型胶原基因敲除小鼠和 DEB 患者的角质形成细胞和成纤维细胞的活动性较低，但添加重组 VII 型胶原可以挽救受损的活动性。此外，重组VII型胶原蛋白应用于皮肤溃疡可改善伤口愈合，我们建议将VII型胶原蛋白作为抗溃疡药物。因此，我们认为我们可以建立一种使用重组VII型胶原蛋白治疗伤口的新策略。

项目成果

Keratotic lesions in epidermolysis bullosa simplex with mottled pigmentation.

单纯性大疱性表皮松解症的角化病变伴有斑驳色素沉着。

• 发表时间: 2005
• 期刊: J Dermatol Sci 52
• 作者: Toki T;Katsuoka F;Ito E et al.;J.Sasaki;Nakamura H;T.Wada;Yasukawa K
• 通讯作者: Yasukawa K

Ultraviolet A-induced Production of Matrix Metalloproteinase-1 Is Mediated by Macrophage Migration Inhibitory Factor (MIF) in Human Dermal Fibroblasts*

• DOI: 10.1074/jbc.m303650200
• 发表时间: 2004-01
• 期刊: Journal of Biological Chemistry
• 影响因子: 4.8
• 作者: Hirokazu Watanabe;T. Shimizu;J. Nishihira†;R. Abe;T. Nakayama;M. Taniguchi;H. Sabe;T. Ishibashi;H. Shimizu

Tomita Y, et al.: "Simultaneous occurrence of three squamaous cell carcinomas in a recessive dystrophic epidermolysis bullosa patient"Acta Derm Venereol. 83. 225-226 (2003)

Tomita Y 等人：“隐性营养不良性大疱性表皮松解症患者同时发生三种鳞状细胞癌”Acta Derm Venereol。

The G2028R glycine substitution mutation in COL7A1 leads to marked inter-familiar clinical heterogeneity in dominant dystrophic epidermolysis bullosa.

COL7A1 中的 G2028R 甘氨酸取代突变导致显性营养不良性大疱性表皮松解症中明显的临床异质性。

• 发表时间: 2004
• 期刊: Arch Dermatol Res 295
• 作者: Toki T;Katsuoka F;Ito E et al.;J.Sasaki;Nakamura H;T.Wada;Yasukawa K;K.Reif;Sawamura D;H.Watanabe;Nakamura H
• 通讯作者: Nakamura H

Siegel DH, et al.: "Loss of kindlin-1, a human homolog of the Caenorhabditis elegans actin-extracellular-matrix linker protein UNC-112, causes Kindler syndrome."Am J Hum Genet. 73. 174-187 (2003)

Siegel DH 等人：“kindlin-1（秀丽隐杆线虫肌动蛋白细胞外基质连接蛋白 UNC-112 的人类同源物）的丢失会导致 Kindler 综合征。”Am J Hum Genet。