Inhibition of proliferative vitreo retinopathy by controling the related transcription factors.

通过控制相关转录因子抑制增殖性玻璃体视网膜病变。

• 批准号: 09671797
• 负责人: TAKAHASHI Masayo
• 金额: $ 1.92万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671797/
• 关键词: PVR; NF_<kappa>B; E2F; gene transfer; NFkBデコイ; 網膜芽細胞腫; 変異型Rb遺伝子導入; 遺伝子治療; NFkB転写因子

Purpose. Previous studies suggested that proliferation and migration of retinal pigment epithelial cells are deeply involved in pathogenesis of proliferative vitreo retinopathy (PVR). Recently it is reported that decoys, double-stranded phosphorothioate oligonucleotides, have the same sequence of binding site of transcription factors. In this study we tested the effect of E2F decoys and NFkappaB decoys in the proliferation of cultured human retinal pigment epithelial (RPE) cells and in vivo animal model of PVR.Methods. HVJ cationic liposomes were prepared by replacing phosphatidylserine of HVJ liposomes (Hangai, et al, 1996) to DC cholesterol. Results. It was shown that E2F decoys highly combined to nuclear extracts from human fibroblast cells and NFkappaB decoys also combined to nuclear extracts from human RPE cells stimulated by interleukin 1beta (IL-1beta). RT-PCR indicated that E2F decoys introduced into human fibroblast decreased expression of important factor that is regulating cell cycle, it showed that NFkappaB decoys introduced into human RPE cells reduced transcripts of IL-1beta in IL-1beta stimulated RPE cells. BrdU labeling index and DNA synthesis (H-thymidine uptake) clarified that cell proliferation was highly inhibited with E2F decoys. Cells on culture dishes with NFkappaB decoys were significantly inhibited cell migrations compared with scrambled decoys. In vivo experiment NFkappaB decoys, transferred by HVJ cationic liposomes into human cultured fibroblast, were injected in the vitreous cavity of rabbit PVR model. The appearances of rabbit PVR were scored by a classification of Blumenkraz et al. The incidences of PVR were not significantly different between with NFkappaB decoys and with scrambled decoys. Conclusions. These results suggest that E2F decoys and NFkappaB decoys have the potential to treat proliferative vitreoretinopathy in vitro and it is needed more study to vitro use.

目的。既往研究表明，视网膜色素上皮细胞的增殖和迁移与增殖性玻璃体视网膜病变（PVR）的发病机制密切相关。最近有报道称，诱饵双链硫代磷酸寡核苷酸具有相同的转录因子结合位点序列。在本研究中，我们测试了 E2F 诱饵和 NFkappaB 诱饵对培养的人视网膜色素上皮 (RPE) 细胞和 PVR 体内动物模型增殖的影响。方法。通过将HVJ脂质体(Hangai等人，1996)的磷脂酰丝氨酸替换为DC胆固醇来制备HVJ阳离子脂质体。结果。结果表明，E2F 诱饵与人成纤维细胞的核提取物高度结合，NFkappaB 诱饵也与白细胞介素 1β (IL-1β) 刺激的人 RPE 细胞的核提取物结合。 RT-PCR表明，引入人成纤维细胞的E2F诱饵降低了调节细胞周期的重要因子的表达，表明引入人RPE细胞的NFkappaB诱饵降低了IL-1β刺激的RPE细胞中IL-1β的转录本。 BrdU 标记指数和 DNA 合成（H-胸苷摄取）表明 E2F 诱饵高度抑制细胞增殖。与乱序诱饵相比，含有 NFkappaB 诱饵的培养皿上的细胞显着抑制细胞迁移。体内实验将NFkappaB诱饵通过HVJ阳离子脂质体转移到人培养的成纤维细胞中，注射到兔PVR模型的玻璃体腔中。兔子 PVR 的外观按照 Blumenkraz 等人的分类进行评分。 NFkappaB 诱饵和乱序诱饵之间的 PVR 发生率没有显着差异。结论。这些结果表明E2F诱饵和NFkappaB诱饵具有体外治疗增殖性玻璃体视网膜病变的潜力，并且需要更多的体外使用研究。

项目成果

M Akimoto: "Growth inhibition of cultured human Tenon's fibroblastic cells by targeting the E2F transcription Factor." Exp.Eye Res. 67. 395-401 (1998)

M Akimoto：“通过靶向 E2F 转录因子抑制培养的人 Tenon 成纤维细胞的生长。”

Akimoto,: "Growth inhibition of cultured human Tenon's fibroblastic cells by targeting the E2F transcription Factor." Exp. Eye Res.67. 395-401 (1998)

Akimoto：“通过靶向 E2F 转录因子抑制培养的人 Tenon 成纤维细胞的生长。”

Akimoto M: "Adenovirally expressed basic fibroblast growth factor rescues photoreceptor cells in RCS rats." Invest Ophthalmol Vis Sci. 40 (2). 273-9 (1999)

Akimoto M：“腺病毒表达的碱性成纤维细胞生长因子可以拯救 RCS 大鼠的感光细胞。”