Constitutive expression of phVEGF165 after intramuscular gene transfer promotes collateral vessel development in patients with critical limb ischemia
Constitutive expression of phVEGF165 after intramuscular gene transfer promotes collateral vessel development in patients with critical limb ischemia
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DOI：
10.1161/01.cir.97.12.1114
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发表时间：
1998-03-31
期刊：
影响因子：
37.8
通讯作者：
Isner, JM
中科院分区：
文献类型：
--
作者：
Baumgartner, I;Pieczek, A;Isner, JM
背景 - 临床前研究表明，血管生成生长因子可以刺激侧支动脉的发育，这一概念称为“治疗性血管生成”。该 1 期临床试验的目的是 (1) 证明使用编码内皮细胞有丝分裂原的裸质粒 DNA 进行肌肉内基因转移的安全性和可行性，以及 (2) 分析对严重肢体缺血患者的潜在治疗益处。 方法和结果 - 在 9 名患有不愈合缺血性溃疡 (n = 7/10) 和/或由于外周原因引起的静息痛 (n = 10/10) 的患者的 10 条肢体中进行了基因转移。动脉疾病。将总剂量为4000μg的编码人血管内皮生长因子(phVEGF(165))的165个氨基酸亚型的裸露质粒DNA直接注射到缺血肢体的肌肉中。通过 ELISA 监测的 VEGF 血清水平短暂增加来记录基因表达。踝臂指数显着改善（0.33 +/- 0.05 至 0.48 +/- 0.03，P = .02）；通过造影血管造影直接记录 7 个肢体新可见的侧支血管；磁共振血管造影显示 8 个肢体远端血流改善的定性证据。 7 条肢体中有 4 条的缺血性溃疡愈合或显着改善，其中 3 名建议进行膝下截肢的患者成功保肢。基因治疗 10 周后从截肢者获得的组织样本通过免疫组织化学显示有增殖的内皮细胞灶。 PCR 和 Southern blot 分析表明，少量质粒 DNA 持续存在，6 名患者的并发症仅限于短暂的下肢水肿，这与 VEGF 增强血管通透性的情况一致。结论 - 这些发现可以谨慎解释，表明肌内注射裸露质粒 DNA 可以实现 VEGF 的组成型过度表达，足以在选定的严重肢体缺血患者中诱导治疗性血管生成。
Background - Preclinical studies have indicated that angiogenic growth factors can stimulate the development of collateral arteries, a concept called "therapeutic angiogenesis." The objectives of this phase 1 clinical trial were (1) to document the safety and feasibility of intramuscular gene transfer by use of naked plasmid DNA encoding an endothelial cell mitogen and (2) to analyze potential therapeutic benefits in patients with critical limb ischemia.Methods and Results - Gene transfer was performed in 10 limbs of 9 patients with nonhealing ischemic ulcers (n = 7/10) and/or rest pain (n = 10/10) due to peripheral arterial disease. A total dose of 4000 mu g of naked plasmid DNA encoding the 165-amino-acid isoform of human vascular endothelial growth factor (phVEGF(165)) was injected directly into the muscles of the ischemic limb. Gene expression was documented by a transient increase in serum levels of VEGF monitored by ELISA. The ankle-brachial index improved significantly (0.33 +/- 0.05 to 0.48 +/- 0.03, P = .02); newly visible collateral blood vessels were directly documented by contrast angiography in 7 limbs; and magnetic resonance angiography showed qualitative evidence of improved distal flow in 8 limbs. Ischemic ulcers healed or markedly improved in 4 of 7 limbs, including successful limb salvage in 3 patients recommended for below-knee amputation. Tissue specimens obtained from an amputee 10 weeks after gene therapy showed foci of proliferating endothelial cells by immunohistochemistry. PCR and Southern blot analyses indicated persistence of small amounts of plasmid DNA, Complications were limited to transient lower-extremity edema in 6 patients, consistent with VEGF enhancement of vascular permeability.Conclusions - These findings may be cautiously interpreted to indicate that intramuscular injection of naked plasmid DNA achieves constitutive overexpression of VEGF sufficient to induce therapeutic angiogenesis in selected patients with critical limb ischemia.