Relationship between periodontal disease and systemic diseases: molecular mechanisms and novel therapy discovery

牙周病与全身性疾病的关系：分子机制和新疗法的发现

• 批准号: 22390350
• 负责人: YAMAMOTO Kenji
• 金额: $ 12.31万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22390350/
• 关键词: 感染症; 酵素; 細菌; 歯学; 薬学

To establish the molecular basis of the possible linkage between periodontal diseases and a variety of systemic diseases such as atherosclerosis, obesity, diabetes, and preterm birth and fetal death, we used various animal models and different cell types and manipulated the endolysosomal aspartic proteinase cathepsin E (CatE) as a host-derived protease and gingipains (GP) as a periodontopathogen-derived protease. In this study, we first found that CatE plays a crucial role in host defense against infection with Porphyromonas gingivalis, a major etiological bacterium of adult periodontal disease through proper trafficking and cell surface expressionof Toll-like receptors such as TLR4 in macrophages. Second, GP mediates atherosclerosis progression accelerated by P. gingivalisinfection through selective proteolysis of apolipoprotein B-100 in LDL cholesterol. Third,GP acts as a strong virulence factor to induce preterm birth and low birth weight through the enhancement of immune responses to pregnant mice infected with P. gingivalis. Fourth, CatE plays a crucial role in normal development of adipose tissues through biochemical analysis of CatE-/-mice that were fed a high-fat diet as an obesity mouse model. Forth, using techniques of cDNA display, selection-by-function, γ-ligation-based block shuffling and others, we generated peptide inhibitors for CatE and GP. The newly developed inhibitors for each enzyme showed an IC50 of nM order and high selectivity. This method is thus expected to be widely applicable for the development of peptide inhibitors of various proteases.

为了建立牙周病和各种系统性疾病如动脉粥样硬化、肥胖、糖尿病、早产和胎儿死亡之间可能联系的分子基础，我们使用各种动物模型和不同的细胞类型，并操纵作为宿主来源的蛋白酶的内溶酶体天冬氨酸蛋白酶组织蛋白酶E（CatE）和作为牙周病病原体来源的蛋白酶的牙龈卟啉酶（GP）。在本研究中，我们首次发现CatE在宿主防御牙龈卟啉单胞菌感染中起着至关重要的作用，牙龈卟啉单胞菌是成人牙周病的主要病原菌，通过适当的运输和细胞表面表达Toll样受体，如TLR 4在巨噬细胞中。第二，GP通过选择性水解LDL胆固醇中的载脂蛋白B-100介导牙龈卟啉单胞菌感染加速的动脉粥样硬化进展。第三，GP作为一种强毒力因子，通过增强感染牙龈卟啉单胞菌的孕鼠的免疫应答，诱导早产和低出生体重。第四，通过对作为肥胖小鼠模型的喂食高脂肪饮食的CatE-/-小鼠的生化分析，CatE在脂肪组织的正常发育中起关键作用。第四，利用cDNA展示、功能选择、基于γ-连接的区块改组等技术，筛选出了CatE和GP的抑制肽。新开发的每种酶的抑制剂显示出nM量级的IC 50和高选择性。因此，该方法有望广泛应用于各种蛋白酶的肽抑制剂的开发。

项目成果

カテプシンEによるガン細胞の増殖抑制とアポトーシス誘導および関連シグナリング

组织蛋白酶 E 及相关信号传导抑制癌细胞生长并诱导细胞凋亡

• 发表时间: 2012
• 作者: 山本健二;川久保友世;安河内篤;筑波隆幸
• 通讯作者: 筑波隆幸

ヒト卵胞液中のcathepsin Eの存在と意義

人卵泡液中组织蛋白酶 E 的存在及其意义

• 发表时间: 2010
• 作者: 後藤志信、尾崎康彦;他8名
• 通讯作者: 他8名

Emerging roles of cathepsin E in host defense mechanisms.

• DOI: 10.1016/j.bbapap.2011.05.022
• 发表时间: 2012
• 期刊: Biochimica et biophysica acta
• 作者: Kenji Yamamoto;Tomoyo Kawakubo;A. Yasukochi;T. Tsukuba

Idetification of two transcripts and in vivo promoter analysis for cathepsin E

组织蛋白酶 E 的两个转录本的鉴定和体内启动子分析

• 发表时间: 2012
• 作者: Okamoto K;Sakai E;Nishishita K;Yamamoto K;Tsukuba T.
• 通讯作者: Tsukuba T.

DMBA/TPA二段階乳頭腫形成モデルにおける表皮カテプシンEの役割

表皮组织蛋白酶 E 在 DMBA/TPA 两阶段乳头状瘤形成模型中的作用

• 发表时间: 2011
• 作者: 安河内篤;川久保友世;中村誠司;山本健二
• 通讯作者: 山本健二