Inhibition of ischemia induced-apoptosis in cardiac myocytes by ATF3 and its implication in gene therapy

ATF3抑制心肌细胞缺血诱导的凋亡及其在基因治疗中的意义

• 批准号: 13670696
• 负责人: ITO Hiroshi
• 金额: $ 2.24万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670696/
• 关键词: ATF3; doxorubicin; myocardial ischemia; reperfusion; c-jun; AP-1; p53; アポトーシス; アデノウイルスベクター

ATF3 (activating transcription factor 3) is a member of the ATF/CREB family, which binds to CRE and AP-1 consensus sequences. We have recently reported that ATF3 inhibits apoptosis of cultured neonatal rat cardiac myocytes induced by DOX. ATF3 was immediately induced at transcriptional and translational level by stimulation of DOX, peaking at 1 hour and at 3 hours respectively. To extend our understanding of the physiological functions of ATF3 in the DOX-induced apoptosis, we produced the adenovirus vector containing coding sequences of ATF3 (AdATF3) and directed overexpression of ATF3 in cardiac myocytes. The inhibition of DOX-induced apoptosis by AdATF3 was shown by flow cytometry, cefi viabih'ty assay and TUNEL staining analyzes. We further demonstrated that ATF3 formed heterodimer with c-Jun both in the cells with or without DOX treatment, suggesting that inappropriate formation of hetero- and homodimer of ATF3 may result in the inhibition of apoptosis in cardiac myocytes with AdATF3.We further investigated AdATF3 can inhibit apoptosis induced by ischemia in cardiac myocytes. AdATF3 inhibited apoptosis induced by hypoxic stimulation in vitro. We also showed that AdATF3 inhibited apoptosis induced by ischemia-reperfusion.In conclusion, these results indicate that overexpression of ATF3 inhibits apoptosis in cardiac myocytes, and suggest a cardio protective role of ATF3. Our data may open a new avenue for gene therapy against myocardial damage by a variety of cardiovascular diseases.

ATF3(激活转录因子3)是ATF/CREB家族的成员，与Cre和AP-1的共同序列结合。我们最近报道了ATF3抑制DOX诱导的培养乳鼠心肌细胞的凋亡。DOX刺激即刻在转录和翻译水平诱导ATF3，分别在1h和3h达到高峰。为了进一步了解ATF3在DOX诱导的细胞凋亡中的生理功能，我们构建了含有ATF3编码序列的腺病毒载体(AdATF3)，并在心肌细胞中定向过表达ATF3。流式细胞仪、头孢菲法和TUNEL染色分析显示AdATF3对DOX诱导的细胞凋亡有抑制作用。我们进一步证明，无论有无DOX处理，ATF3都与c-jun形成异源二聚体，提示ATF3异源二聚体和同源二聚体的不适当形成可能导致AdATF3抑制心肌细胞的凋亡。AdATF3在体外可抑制缺氧刺激诱导的细胞凋亡。我们还发现AdATF3抑制了缺血再灌流诱导的细胞凋亡。综上所述，这些结果表明ATF3的过表达抑制了心肌细胞的凋亡，提示ATF3具有心脏保护作用。我们的数据可能为针对多种心血管疾病造成的心肌损伤的基因治疗开辟一条新的途径。

项目成果

Tamamori-Adachi M, Ito H. Nobori K, Hayashida K, Kawauchi J, Adachi S, Ikeda M, Kitajima S: "Expression of cyclin Dl and CDK4 causes hypertrophic growth of cardiomyocytes in culture a possible implication for cardiac hypertrophy"Biochem. Biophys. Res. Com

Tamamori-Adachi M、Ito H. Nobori K、Hayashida K、Kawauchi J、Adachi S、Ikeda M、Kitajima S：“细胞周期蛋白 D1 和 CDK4 的表达导致培养物中心肌细胞肥大性生长，可能暗示心脏肥大”Biochem。

Nobori K, Ito H, Tamamori-Adachi M, Adachi S, Ono Y, et al.: "ATF3 Inhibits Doxorubicin-induced Apoptosis in Cardiac Myocytes ; A Novel Cardiorotective Role of ATF3"J. Mol. Cell Cardiol. (in press). (2002)

Nobori K、Ito H、Tamamori-Adachi M、Adachi S、Ono Y 等人：“ATF3 抑制多柔比星诱导的心肌细胞凋亡；ATF3 的新型心脏保护作用”J。

Tamanori-Adachi M, Ito H, S Piyamas, Adachi S, Hiroe M, Shimizu M, Kawauchi J, Sunamori M, Marumo F, Kitajima S, Ikeda M Tamamori-Adachi M, Ito H, S Piyamas et al.: "Critical role of Cyclin Dl nuclear import in cardiomyocyte proliferation"Circ Res.. 92. 1

Tamanori-Adachi M、Ito H、S Piyamas、Adachi S、Hiroe M、Shimizu M、Kawauchi J、Sunamori M、Marumo F、Kitajima S、Ikeda M Tamamori-Adachi M、Ito H、S Piyamas 等人：“关键

Nozato T, Ito H, Watanabe M, Ono Y Adachi S, Tanaka H, et al.: "Overexpression of cdk inhibitor p16^<INK4a>by adenovirus vector inhibits cardiac hypertrophy in vitro and in vivo : a novel strategy for therapy of cardiac hypertrophy"J. Mol. Cell Cardiol..

Nozato T、Ito H、Watanabe M、Ono Y Adachi S、Tanaka H 等人：“腺病毒载体过表达 cdk 抑制剂 p16^<INK4a> 可在体外和体内抑制心脏肥大：一种治疗心脏病的新策略

Tamamori-Adachi M, Ito H, Nobori K 他: "Expression of cyclin D1 and CDK4 causes hypertrophic growth of cardiomyocytes in cuture : a possible implication for cardiac hypertrophy"Biochem. Biophys. Res Commun.. 296(2). 274-280 (2002)

Tamamori-Adachi M、Ito H、Nobori K 等人：“细胞周期蛋白 D1 和 CDK4 的表达导致心肌细胞肥大生长：心脏肥大的可能暗示”Biochem Res Commun. 296(2)。 274-280 (2002)