Endograft technology: a delivery vehicle for intravascular gene therapy.
Endograft technology: a delivery vehicle for intravascular gene therapy.
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内移植技术：血管内基因治疗的递送载体。
DOI：
10.1016/j.jvs.2003.11.033
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发表时间：
2004
影响因子：
4.3
通讯作者：
Livingstone,Alan
中科院分区：
文献类型：
--
作者：
Eton,Darwin;Yu,Hong;Wang,Yingcai;Raines,Jeffrey;Striker,Gary;Livingstone,Alan
目的：观察血管平滑肌细胞（SMC）在体内7个月内是否保留和表达逆转录病毒转导的基因。方法：用逆转录病毒转导犬颈静脉平滑肌细胞，导入组织型纤溶酶原激活物（t-PA）和β-半乳糖苷酶基因。然后将这些细胞充满到新型双层涤纶移植物中并培养36至48小时。将移植物安装在Palmaz支架上，并在SMC供体犬（n = 6）的肾下主动脉中球囊扩张。移植物在1、2、3、4、5和7个月时回收。将充满仅用β-半乳糖苷酶基因转导的SMC的对照覆膜支架置于犬中，并在2、3和4个月时恢复移植物。用酶联免疫吸附试验分析t-PA抗原的浓度和表达，用X-gal染色鉴定移植物、新生内膜和下面的主动脉中保留的工程SMC（蓝色核）。从移植物中回收的SMC的t-PA抗原浓度和t-PA活性在实验期间保持升高（7个月），（3.7 ± 0.2 ng/mL/105个细胞/24小时和1.4 ± 0.1 IU/mL/105个细胞/24小时）（0.5 ± 0.03 ng/mL/105个细胞/24小时和0.07 ± 0.00 IU/mL/105个细胞/24小时; P <0.001）。没有移植物狭窄observed. CONCLUSION逆转录病毒工程血管平滑肌细胞存活的植入创伤，重新填充每个移植物，迁移到下面的主动脉，并表达了7个月的实验持续时间的转导基因。这种双层涤纶覆膜支架模型为研究直接血管内基因治疗提供了一个平台。
PURPOSEThe purpose of this study was to determine whether vascular smooth muscle cells (SMCs) suffused into a bilayered stent graft retain and express a retrovirally transduced gene for 7 months in vivo.METHODSSMCs harvested from dog jugular vein were retrovirally transduced to introduce genes for tissue plasminogen activator (t-PA) and β-galactosidase. These cells were then suffused into a novel dual-layered Dacron graft and cultured for 36 to 48 hours. The grafts were mounted on a Palmaz stent and balloon- expanded in the infrarenal aorta of the SMC donor dogs (n = 6). Grafts were recovered at 1, 2, 3, 4, 5, and 7 months. A control endograft suffused with SMCs transduced with only the β-galactosidase gene was placed in the dogs with grafts recovered at 2, 3, and 4 months. t-PA antigen concentration and expression were analyzed with an enzyme-linked immunosorbent assay.RESULTSRetained engineered SMCs (blue nuclei) were identified in the explanted grafts, neointima, and underlying aorta with X-gal staining. The t-PA antigen concentration and t-PA activity from the SMCs recovered from the grafts remained elevated for the duration of the experiment (7 months) at levels significantly higher (3.7 ± 0.2 ng/mL per 105cells per 24 hours and 1.4 ± 0.1 IU/mL per 105cells per 24 hours) than in control endografts (0.5 ± 0.03 ng/mL per 105cells per 24 hours and 0.07 ± 0.00 IU/mL per 105cells per 24 hours; P < .001). No graft stenosis was observed.CONCLUSIONRetrovirally engineered vascular SMCs survived the implantation trauma, repopulated each graft, migrated into the underlying aorta, and expressed the transduced genes for the 7-month duration of the experiment. This bilayered Dacron endograft model provides a platform to study direct intravascular gene therapy.