Analysis of transcriptional control mechanism for the expression of type I collagen and related gene in fibrotic skin disorders

纤维化皮肤病中I型胶原及相关基因表达的转录控制机制分析

• 批准号: 14570799
• 负责人: HATAMOCHI Atsushi
• 金额: $ 1.28万
• 依托单位: Dokkyo University School of Medicine (2003-2004)Chiba University (2002)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570799/
• 关键词: type I collagen; transcription; fibrotic skin diseases; Co1F1; type III collagen; Erythromycin analog; マクロライド誘導体; コラゲナーゼ; 転写因子; Ehlers-Danlos症候群

Fibrotic skin diseases such as systemic sclerosis(SSc) are characterized by excessive accumulation of collagen in the skin and a variety of internal organs. The accumulation of collagen is thought to result from enhanced transcription of collagen in fibroblasts. Collagen type I, a most abundant protein in the dermis, consists of two α1(I) chain and one α2(I) chain which are coordinately expressed. Co1F1, a DNA binding protein which specifically binds to a segment of the α 2(I)collagen promoter at -400bp upstream of the start of transcription, activates transcription of the α2(I)collagen gene in vitro. We investigated on the partial sequences of polypeptide and the cDNA cloning of this transcriptional factor of the α2(T)collagen gene. The protein purified by sequence-specific DNA affinity chromatography were found to consist of 42kDa and 40.5 kDa polypeptides. Sequences of peptide fragments from 42kDa polypeptide were identical to Purα, which is a nuclear protein which has been reported … More to binds to a upstream region of human c-myc gene. Some of those from 40kDa polypeptide were identical to Purβ, has been partially sequenced and has regions of strong homology to Purα. Full length of Purβ cDNA were sequenced. The deductive amino acid sequences of Purα and Purβ showed 61.4% homology. None of treatment modalities for patients with SSc has brought satisfactory improvement. We, therefore, would like to present effects of the Erythromycin analog EM703 on collagen transcription in normal and scleroderma fibroblasts. EM703 reduced collagen production(to 40% in maximum) and mRNA levels of α1(I) collagen(to 30% in maximum) in a dose dependent manner in normal fibroblasts. EM703 markedly reduced those in all 9 SSc fibroblasts. COL1A1 transcription was down-regulated by 30% in the Luciferase assay. Nuclear extracts from fibroblasts treated with EM703 reduced the CCAAT-binding factor(CBF) binding activity, whereas SP1 binding activity was unchanged. These results suggest that EM703 reduce the type I collagen transcription not only in normal but also in SSc fibroblasts and that the CBF is involved in this reduced expression. Less

纤维化皮肤病如系统性硬化症（SSc）的特征在于胶原蛋白在皮肤和各种内脏器官中的过度积累。胶原蛋白的积累被认为是由成纤维细胞中胶原蛋白的转录增强引起的。Ⅰ型胶原是真皮中含量最丰富的蛋白质，由两条α1（Ⅰ）链和一条α2（Ⅰ）链组成，它们协同表达。Co 1F 1是一种DNA结合蛋白，它特异性地结合于α 2（I）胶原基因启动子上游-400bp处，在体外激活α2（I）胶原基因的转录。本研究对α2（T）胶原基因的部分多肽序列进行了测定，并克隆了该转录因子的cDNA。序列特异性DNA亲和层析纯化的蛋白质由42 kDa和40.5kDa的多肽组成。42 kDa多肽的肽段序列与已报道的核蛋白Purα完全一致 ...更多信息 与人c-myc基因的上游区结合。40 kDa多肽中的部分序列与Purβ相同，已部分测序，并与Purα有很强的同源性。测定了Purβ cDNA的全长序列。推导的Purα和Purβ氨基酸序列同源性为61.4%。SSc患者的治疗方式都没有带来令人满意的改善。因此，我们想介绍红霉素类似物EM 703对正常和硬皮病成纤维细胞中胶原蛋白转录的影响。在正常成纤维细胞中，EM 703以剂量依赖性方式降低胶原蛋白产生（最大至40%）和α1（I）胶原蛋白的mRNA水平（最大至30%）。EM 703显著降低所有9个SSc成纤维细胞中的这些。在荧光素酶测定中，COL 1A 1转录下调30%。EM 703处理的成纤维细胞的核提取物降低了CCAAT结合因子（CBF）的结合活性，而SP1结合活性不变。这些结果表明，EM 703不仅在正常成纤维细胞中而且在SSc成纤维细胞中减少I型胶原蛋白转录，并且CBF参与这种减少的表达。少

项目成果

Ehlers-Danlos syndrome type IV with few extrathoracic finding : a newly recognized point mutation in the COL3A1 gene.

Ehlers-Danlos 综合征 IV 型，很少有胸腔外发现：COL3A1 基因中新发现的点突变。

• 发表时间: 2002
• 期刊: Eur Respir J 19・1
• 作者: Kakimura T;Saeki H;et al.;Watanabe A et al.
• 通讯作者: Watanabe A et al.

Akira Watanabe: "Ehlers-Danlos syndrome type IV with few extrathoracic findings : a new recognized point mutation in the COL3A1 gene"Eur Respir J. 19(1). 195-198 (2002)

Akira Watanabe：“几乎没有胸腔外发现的 IV 型 Ehlers-Danlos 综合征：COL3A1 基因中新识别的点突变”Eur Respir J. 19(1)。

Evaluation of functional disability using the health assessment questionnaire in Japanese patients with systemic sclerosis

使用健康评估问卷评估日本系统性硬化症患者的功能障碍

• 发表时间: 2003
• 期刊: J Rheumatol 30・6
• 作者: Kuwana M et al.

Ehlers-Danlos syndrome type IV with few extrathoracic findings: a newly recognized point mutation in the COL3A1 gene

• DOI: 10.1183/09031936.02.00219202
• 发表时间: 2002-01-01
• 期刊: EUROPEAN RESPIRATORY JOURNAL
• 影响因子: 24.3
• 作者: Watanabe, A;Kawabata, Y;Kuriyama, T
• 通讯作者: Kuriyama, T

Ehlers-Danlos Syndrome type IV with few extrathoracic findings : a newly recongnized point mutation in COL3A1 gene.

几乎没有胸腔外发现的 IV 型埃勒斯-当洛斯综合征：COL3A1 基因中新识别的点突变。

• 发表时间: 2002
• 期刊: Eur Reapir J 19・1
• 作者: Hattori N;Komine M;et al.;Watanabe A et al.
• 通讯作者: Watanabe A et al.