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Novel gene therapy of keloid by using siRNA

使用 siRNA 治疗瘢痕疙瘩的新型基因疗法

基本信息

批准号：
16591791
负责人：
SASAKI Satoru
金额：
$ 2.3万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591791/
关键词：
keloid siRNA TGF-β1 MIF gene therapy PGE_2 cAMP

项目摘要

We hypothesized that gene therapy by using small interfering RNA (siRNA) would be valid on keloid. Firstly, we tried to knockdown TGF- 1 mRNA by transduction of siRNA. As a result, TGF- 1 production was down-regulated. We found that collgen production and cell growth in keloid-derived fibroblasts (KF) down-regulated by decreased TGF- 1 production. Therefore we believe that gene therapy by using siRNA may be useful for the treatment of keloid.In this study, We investigated the metabolism of arachidonic acid in normal skin-derived fibroblasts (NF) as well as in keloid-derived fibroblasts (KF) in response to macrophage migration inhibitory factor (MIF), a pluripotent cytokine. We found that MIF enhanced cyclooxygenase-2 (COX-2) activity in NF more than in KF. Consistent with this finding, prostaglandin E_2 (PGE_2), an antifibrogenic molecule, was more significantly increased in NF than in KF by MIF treatment. As regarding E prostanoid receptor 2 (EP2), the level of expression was significantly lower in KF than in NF. On the other hand, Forskolin, a direct activator of adenylcyclase, decreased collagen synthesis in both NF and KF, which indicates that cAMP plays an important role in regulating collagen synthesis. Since PGE_2 induces cAMP production, it is conceivable that increased collagen synthesis in KF might be due to decreased PGE_2 and cAMP production. These findings may aid in the development of a therapeutic strategy for the regulation of collagen synthesis in keloid fibroblasts.

我们假设使用小干扰RNA(SiRNA)对瘢痕疙瘩进行基因治疗是有效的。首先，我们尝试通过siRNA的转导来敲除转化生长因子-1的mRNA。结果，转化生长因子-1的产生被下调。我们发现瘢痕疙瘩成纤维细胞(KF)的胶原生成和细胞生长受转化生长因子-1(TGF-1)生成减少的影响。因此，我们认为使用siRNA进行基因治疗可能对治疗瘢痕疙瘩有用。在本研究中，我们研究了多能细胞因子巨噬细胞移动抑制因子(MIF)对正常皮肤成纤维细胞(NF)和瘢痕疙瘩成纤维细胞(KF)中花生四烯酸代谢的影响。我们发现MIF对NF环氧合酶-2(COX-2)活性的促进作用强于KF。与此相一致的是，经MIF治疗后，抗纤维化分子前列腺素E_2(PGE_2)在NF组较KF组显著升高。E前列腺素受体2(EP2)在KF中的表达水平显著低于NF。另一方面，腺苷环化酶的直接激活剂Forskolin减少了Nf和KF的胶原合成，表明cAMP在调节胶原合成中起重要作用。由于PGE_2可诱导cAMP的产生，推测KF胶原合成的增加可能是由于PGE_2和cAMP的减少所致。这些发现可能有助于开发一种治疗策略来调节瘢痕疙瘩成纤维细胞的胶原合成。