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.

为了建立牙周疾病与动脉粥样硬化、肥胖、糖尿病、早产和胎儿死亡等多种全身性疾病之间可能联系的分子基础，我们使用了不同的动物模型和不同的细胞类型，并操纵内溶酶体天冬氨酸蛋白酶组织蛋白酶E(CATE)作为宿主来源的蛋白酶，牙周疼痛(GP)作为牙周病原体来源的蛋白酶。在这项研究中，我们首次发现Cate通过在巨噬细胞中适当运输和细胞表面表达Toll样受体如TLR4，在宿主防御牙龈卟啉单胞菌感染中发挥关键作用。第二，GP通过选择性地分解低密度脂蛋白中的载脂蛋白B-100来介导牙龈假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，通过增强感染牙龈假单胞菌的孕鼠的免疫反应，导致早产和低出生体重。第四，通过对喂食高脂饮食的Cate-/-小鼠进行生化分析，作为肥胖小鼠模型，Cate在脂肪组织的正常发育中发挥了关键作用。第四，利用基因展示、功能选择、基于γ连接的区块改组等技术，我们合成了CATE和GP的多肽抑制剂。新开发的每种酶的抑制剂都显示出NM量级的IC50和高选择性。因此，该方法有望广泛应用于各种蛋白酶的多肽抑制剂的开发。

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