Preclinical evaluation of inducible nitric oxide synthase lipoplex gene therapy for inhibition of stent-induced vascular neointimal lesion formation
Preclinical evaluation of inducible nitric oxide synthase lipoplex gene therapy for inhibition of stent-induced vascular neointimal lesion formation
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DOI：
10.1089/104303403321208970
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发表时间：
2003-03-01
影响因子：
4.2
通讯作者：
Von der Leyen, HE
中科院分区：
文献类型：
--
作者：
Muhs, A;Heublein, B;Von der Leyen, HE
一些报告已经确立了一氧化氮合酶（NOS）基因转移用于抑制血管损伤后平滑肌细胞（SMC）增殖的概念。为了最大限度地减少与病毒基因转移相关的潜在风险，我们开发了一种基于脂质体的基因转移方法，采用诱导型一氧化氮合酶（iNOS）过表达来抑制支架诱导的新内膜病变形成。使用 Infiltrator 局部药物输送装置将治疗性脂质复合物转移至哥廷根小型猪的股动脉或冠状动脉。通过 iNOS 特异性免疫组织化学分析局部 iNOS 脂质复合物转移的效率。通过血管内超声 (IVUS) 和计算机形态测定法分析 NO 介导的支架诱导的新内膜病变形成的抑制作用。基因转移效率随剂量增加而增加，iNOS 阳性血管面积最大可达 44.3+/- 4.2%（剂量，2 杯 iNOS lipoplex）。转移 2 mg iNOS 脂质复合物可显着抑制内侧 SMC 的增殖细胞核抗原 (PCNA) 表达（免疫组织化学）（111 +/- 27 细胞 [iNOS] 对比 481 +/- 67 细胞[对照；PCNA 阳性内侧细胞]）。 IVUS 分析表明，iNOS 脂质复合物的局部转移导致股骨支架内斑块面积显着减少（对照，40.85 +/- 6.37 mm(2)；iNOS，24.69 +/- 1.8 mm(2)；p = 0.03）。根据组织学形态计量学测定，冠状动脉支架内病变形成减少约 45%（对照，4.0 +/- 0.29；iNOS，2.2 +/- 0.30；p < 0.01）。总之，本研究表明，iNOS 脂质复合物的局部壁内递送可以在抑制支架诱导的新内膜病变形成方面发挥治疗作用。加上这种基因治疗方法的非病毒特征，这些发现可能对基于 NOS 的基因治疗向临床实践的转变产生重要影响。
Several reports have established the concept of nitric oxide synthase ( NOS) gene transfer for inhibiting smooth muscle cell (SMC) proliferation after vascular injury. To minimize potential risks associated with viral gene transfer, we developed a liposome-based gene transfer approach employing inducible NOS ( iNOS) overexpression for inhibition of stent-induced neointimal lesion formation. Therapeutic lipoplexes were transferred to femoral or coronary arteries of Goettingen minipigs, using the Infiltrator local drug delivery device. Efficiency of local iNOS lipoplex transfer was analyzed by iNOS-specific immunohistochemistry. NO-mediated inhibition of stent-induced neointimal lesion formation was analyzed by intravascular ultrasound (IVUS) and computerized morphometry. Gene transfer efficiency increased dose dependently to a maximum of 44.3+/- 4.2% iNOS-positive vessel area (dose, 2 mug of iNOS lipoplex). Proliferating cell nuclear antigen ( PCNA) expression of medial SMCs ( immunohistochemistry) was inhibited significantly by transfer of 2 mg of iNOS lipoplexes (111 +/- 27 cells [iNOS] versus 481 +/- 67 cells [control; PCNA-positive medial cells]). IVUS analysis demonstrated that local transfer of iNOS lipoplexes resulted in a significant reduction of femoral in-stent plaque area (control, 40.85 +/- 6.37 mm(2); iNOS, 24.69 +/- 1.8 mm(2); p = 0.03). Coronary in-stent lesion formation was reduced by about 45% as determined by histologic morphometry ( control, 4.0 +/- 0.29; iNOS, 2.2 +/- 0.30; p< 0.01). In conclusion, this study demonstrates that local intramural delivery of iNOS lipoplexes can exert therapeutic effects in inhibiting stent-induced neointimal lesion formation. Together with the nonviral character of this gene therapy approach, these findings may have important impact on the transition of NOS-based gene therapy to clinical practice.