Joint Study on the Therapy of Diseases by Gene Transfer

基因转移治疗疾病的联合研究

• 批准号: 05044170
• 负责人: KANEDA Yasufumi
• 金额: $ 5.44万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05044170/
• 关键词: HVJ-liposomes; Neointima formation; Antisense oligonucleotides; E2F decoys; NO synthase; Antistenosis; 遺伝子導入; 遺伝子治療; 循環器疾患

We developed efficient gene delivery system called HVJ-liposomes, Using this delivery system, we attempted to prevent restsnosis after angioplasty which is a barrier to the therapy of myocardial infarction. We examined the effect of AS-ODNs for the therapy of restenosis. First we introduced AS-PCNA and AS-cdc2 kinase into balloon injured vessels by HVJ-liposome. Cotransfer of both AS-ODNs resulted in the simultanuous inhibition of PCNA and cdc2 kinase mRNA expression to undetectable levels. Then HVJ-liposome containing 15 uM AS-PCNA and AS-cdc2 kinase was injected into balloon injured rat carotid artery. Neointima formation was completely inhibited by this treatment, whereas the sense-ODNs had no effect. The blockade of neointima formation by these AS continiued for 2 weeks by a single intraluminal administration and the inhibitory effect lasted for a period of 8 weeks after transfection.We further examined effect of AS-ODN against various cell-cycle regulators on neointima formation. … More The combination of AS-PCNA and cdc2 kinase or AS-cdc2 kinase and cdk2 kinase were most effective for the prevention of neointima formation. Then, we developed new strategy to inhibit cell proliferation by the introduction of a single molecule. The transcription of PCNA,cdc2 kinase and some protooncogene were activated by a common transcription factor, E2F,and the consensus DNA sequence recognized by E2F is known to be TTTCGCGC.We investigated the effect of double-stranded ODN designated as competitor of E2F on the prevention of neointima formation. We examined the effect of E2F decoy on the prevention of restenosis. E2F decoy was introduced into balloon injured rat carotid artery by HVJ-liposome. Our results demonstrated a marked suppression of neointima formation at 2 weeks sfter angioplasty by the decoy against E2F.At a dosage of 3 uM,decoy ODN inhibited neointima formation by approximately 80 % compared to vessels treated with HVJ-liposome alone or mismatched decoy ODN-treated vessels. Then, we employed NO synthase cDNA to inhibit neointima formation.NOS introduced into blood walls by HVJ-liposomes inhibited neointima formation for at least one month. These strategies will be able to be applied for the treatment of restenosis in humans. Less

我们开发了一种名为 HVJ-脂质体的高效基因传递系统，利用这种传递系统，我们试图预防血管成形术后的休息，这是心肌梗塞治疗的障碍。我们检查了 AS-ODN 治疗再狭窄的效果。首先，我们通过 HVJ 脂质体将 AS-PCNA 和 AS-cdc2 激酶引入球囊损伤的血管中。两种 AS-ODN 的共转移导致 PCNA 和 cdc2 激酶 mRNA 表达同时抑制至不可检测的水平。然后将含有15 uM AS-PCNA和AS-cdc2激酶的HVJ-脂质体注射到球囊损伤的大鼠颈动脉中。这种处理完全抑制了新内膜的形成，而正义ODN则没有效果。通过单次腔内给药，这些AS对新内膜形成的阻断持续2周，并且抑制作用在转染后持续8周。我们进一步检查了AS-ODN针对各种细胞周期调节剂对新内膜形成的影响。 … 更多 AS-PCNA 和 cdc2 激酶或 AS-cdc2 激酶和 cdk2 激酶的组合对于预防新内膜形成最有效。然后，我们开发了通过引入单分子来抑制细胞增殖的新策略。 PCNA、cdc2激酶和一些原癌基因的转录是由一个共同的转录因子E2F激活的，E2F识别的共有DNA序列为TTTCGCGC。我们研究了被指定为E2F竞争者的双链ODN在预防新内膜形成中的作用。我们检查了 E2F 诱饵对预防再狭窄的效果。通过HVJ-脂质体将E2F诱饵引入球囊损伤的大鼠颈动脉。我们的结果表明，血管成形术后 2 周，针对 E2F 的诱饵明显抑制了新内膜形成。与单独使用 HVJ 脂质体处理的血管或经过错配诱饵 ODN 处理的血管相比，3 uM 剂量的诱饵 ODN 抑制了约 80% 的新内膜形成。然后，我们利用NO合酶cDNA抑制新内膜形成。通过HVJ-脂质体将NOS引入血壁，抑制新内膜形成至少1个月。这些策略将能够应用于人类再狭窄的治疗。较少的

项目成果

R.Morishita: "Novel and effective gene transfer technique for study of vascular renin angiotensin system" Journal of Clinical Investigation. 91. 2580-2585 (1993)

R.Morishita：“用于研究血管肾素血管紧张素系统的新颖有效的基因转移技术”临床研究杂志。

Naruya Tomita: "A novel gene transfer technique mediated by HVJ(Sendai virus),nuclear protein and liposomes." Cancer Diagnosis and Prevention. 18. 485-491 (1994)

Naruya Tomita：“一种由 HVJ（仙台病毒）、核蛋白和脂质体介导的新型基因转移技术。”

Ryuichi Morishita: "Novel molecular strategy "decoys" against E2F binding site inhibits neointimal hyperplasia by intra-luminal approach." Proc.Natl.Acad.Sci.(USA). (in press).

Ryuichi Morishita：“针对 E2F 结合位点的新型分子策略‘诱饵’通过腔内方法抑制新内膜增生。”

R.Morishita: "Novel in vitro gene method for study of local modulators in vascular smooth muscle cells" Hypertension. 21. 894-899 (1993)

R.Morishita：“用于研究血管平滑肌细胞局部调节剂的新型体外基因方法”高血压。

R.Morishita: "Single intraluminal delivery of antisense cdc2・kinase and PCNA oligo-nucleotides results in chronic inhibition of neointimal hyperplasia" Proc.Natl.Acad.Sci.(USA). 90. 8874-8878 (1993)

R.Morishita：“反义 CDC2·激酶和 PCNA 寡核苷酸的单次腔内递送导致新内膜增生的慢性抑制”Proc.Natl.Acad.Sci.(USA) 90. 8874-8878 (1993)。