The development of basic technologies for the cellular therapy of corneal epithelial cells by the regulation of cellular senescence and epigenetic changes

通过调控细胞衰老和表观遗传变化进行角膜上皮细胞治疗的基础技术的发展

• 批准号: 20390451
• 负责人: KINOSHITA Shigeru
• 金额: $ 12.56万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20390451/
• 关键词: 細胞老化; エピジェネティクス; 角膜上皮; 眼表面疾患; 移植再生医療; 移植再生治療; エピジェネティックス; 遺伝子導入; プラスミド; レンチウイルス

In this study, in order to establish the method to transplant corneal epithelial cells which are amplified ex vivo from the limited number of autologous corneal epithelial cells, we investigated the cellular senescence and epigenetic changes of the cultured corneal epithelial cells as well as the possible countermeasures against these culture-associated drawbacks. We have found that the expression of the keratin 12 gene significantly decreased rapidly after the cell dissociation of the corneal epithelial cells, which was found to be due to the activation of several signaling pathways such as MAPK accompanied with the epigenetic changes of the keratin 12 promoter. The proliferation of the corneal epithelial cell was found to be significantly enhanced by the introduction of SV40 large T antigen which is known to inhibit p53 and Rb tumor- suppressive pathways. Quite interestingly, when corneal epithelial stem cells were transfected with the SV40 large T antigen, the endogenous hTERT gene was found to be activated, which is thought to be the risk to lead the tumorigenic transformation of these cells.

在这项研究中，为了建立移植角膜上皮细胞的方法，这些方法是从有限数量的自体角膜上皮细胞中放大的，我们研究了培养的角膜上皮细胞的细胞衰老和表观遗传变化，以及可能针对这些培养的​​培养物相关的缺点。我们发现角蛋白12基因的表达在角膜上皮细胞的细胞解离后迅速迅速降低，这是由于几种信号传导途径的激活，例如MAPK伴随着角质素12启动子的表观遗传变化。通过引入SV40大T抗原，发现角膜上皮细胞的增殖可以显着增强，该抗原已知会抑制p53和RB肿瘤抑制途径。有趣的是，当用SV40大T抗原转染角膜上皮干细胞时，发现内源性HTERT基因被激活，这被认为是引导这些细胞肿瘤性转化的风险。

项目成果

ARVO-JOS Symposium.

ARVO-JOS 研讨会。

• 发表时间: 2009
• 期刊: Jpn J Ophthalmol 54
• 作者: 森実勲;白馬伸洋;兵頭純;暁清文;岸部幹;Kinoshita S.
• 通讯作者: Kinoshita S.

スリット所見で診る角膜疾患80,東京

东京利用狭缝检查诊断出 80 种角膜疾病

• 发表时间: 2008
• 作者: Tamaki Y;Iriyama A;Yanagi Y;木下茂(編集)
• 通讯作者: 木下茂(編集)

Long-Term Phenotypic Study after Allogeneic Cultivated Corneal Limbal Epithelial Transplantation for Severe Ocular Surface Diseases

• DOI: 10.1016/j.ophtha.2010.04.003
• 发表时间: 2010-12-01
• 期刊: OPHTHALMOLOGY
• 影响因子: 13.7
• 作者: Nakamura, Takahiro;Sotozono, Chie;Kinoshita, Shigeru
• 通讯作者: Kinoshita, Shigeru

角膜疾患の未来医療

角膜疾病的未来医疗

• 发表时间: 2009
• 作者: 島友子;青木藍子;福田香織;稲田貢三子;橋本佳子;中島彰俊;長谷川徹;日高隆雄;齋藤滋;木村 茂
• 通讯作者: 木村 茂

Corneal dystrophies

• DOI: 10.1016/j.jfo.2017.05.003
• 发表时间: 2017-06-01
• 期刊: JOURNAL FRANCAIS D OPHTALMOLOGIE
• 影响因子: 1.2
• 作者: Bourges, J. -L.