Joint Study on the Therapy of Diseases by Gene Transfer

基因转移治疗疾病的联合研究

基本信息

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

项目摘要

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

我们开发了称为HVJ-脂质体的有效基因输送系统，使用此输送系统，我们试图防止血管成形术后静息病，这是心肌梗塞治疗的障碍。我们检查了AS-ODN对再狭窄治疗的影响。首先，我们通过HVJ-脂质体将AS-PCNA和AS-CDC2激酶引入了气球受伤的血管中。两种AS-ODN的共转移导致PCNA和CDC2激酶mRNA表达对无法检测到的水平的简单抑制。然后将含有15个UM AS-PCNA和AS-CDC2激酶的HVJ-脂质体注射到球囊受伤的大鼠颈动脉中。这种治疗方法完全抑制了新内膜的形成，而感官odn则没有作用。通过单个腔内给药和抑制作用持续了2周，它们持续了2周，持续了两周，持续了两周。我们进一步检查了AS-ODN对各种细胞周期调节剂对新内膜形成的影响。 …更多的是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，我们制定了通过引入单个分子抑制细胞增殖的新策略。 PCNA，CDC2激酶和某些原子能的转录通过共同的转录因子E2F激活，并且已知通过E2F识别的共有DNA序列是TTTCGCGC。我们研究了指定为E2F竞争者对Neiminimae Neiminima的预防竞争的双链ODN的效果。我们检查了E2F诱饵对预防再狭窄的影响。 E2F诱饵被HVJ脂质体引入了气球受伤的大鼠颈动脉。我们的结果表明，与单独使用HVJ-liposome的血管相比，诱饵对E2F的SSFT血管成形术对NEINSIMA形成显着抑制了SSFT血管成形术。然后，我们不使用合成酶cDNA来抑制新内膜的形成。没有HVJ - 脂质体引入的血墙中，至少抑制了一个月的新内膜形成。这些策略将能够用于治疗人类再狭窄。