RGD peptide-modified adenovirus expressing hepatocyte growth factor and X-linked inhibitor of apoptosis improves islet transplantation.
RGD peptide-modified adenovirus expressing hepatocyte growth factor and X-linked inhibitor of apoptosis improves islet transplantation.
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DOI：
10.1002/jgm.1626
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发表时间：
2011-12
期刊：
影响因子：
--
通讯作者：
Mahato RI
中科院分区：
文献类型：
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作者：
Wu H;Yoon AR;Li F;Yun CO;Mahato RI
胰岛移植具有治疗I型糖尿病的潜力，但其广泛的临床应用受到移植胰岛移植物大量凋亡性细胞死亡和血运重建不良的限制。我们构建了编码人X-连锁凋亡抑制因子和肝细胞生长因子的RGD(Arg-Gly-Asp)基因的表面修饰的腺病毒载体(RGD-AdV-hHGF-hXIAP)。用酶联免疫吸附试验检测外源基因在人胰岛中的表达。将RGD-AdV-hHGF-hXIAP转导的人胰岛移植到链脲佐菌素诱导的糖尿病NOD/SCID小鼠肾被膜下。每周测量小鼠的血糖水平。实验结束时，分离小鼠肾脏，进行免疫荧光染色。RGD基因修饰的腺病毒对人胰岛的转导效率显著提高。病毒转导后HGF和XIAP基因的表达呈剂量依赖性。当暴露在炎性细胞因子的混合物中时，转导RGD-ADV-hHGF-hXIAP的人胰岛显示caspase3活性降低，并减少了细胞凋亡。通过移植RGD-AdV-hHGF-hXIAP转导的人胰岛可以实现长期的正常血糖控制。移植后200天，RGD-AdV-hHGF-hXIAP转导胰岛的肾脏切片免疫荧光染色显示胰岛素和von Willebrand因子(VWF)阳性。这些结果表明，RGD-AdV-hHGF-hXIAP体外转导胰岛可减少胰岛细胞的凋亡性死亡，促进胰岛血运重建，最终可能改善人胰岛移植的预后。
Islet transplantation has the potential for treating type I diabetes; however, its widespread clinical application is limited by the massive apoptotic cell death and poor revascularization of transplanted islet grafts. We constructed a surface-modified adenoviral vector with RGD (Arg-Gly-Asp) sequences encoding human X-linked inhibitor of apoptosis and hepatocyte growth factor (RGD-Adv-hHGF-hXIAP). In vitro transgene expression in human islets was determined by enzyme-liniked immunosorbent assay. RGD-Adv-hHGF-hXIAP-transduced human islets were transplanted under the kidney capsule of streptozotocin-induced diabetic NOD/SCID mice. The blood glucose levels of mice were measured weekly. The kidneys bearing islets were isolated at the end of the experiment and subjected to immunofluorescence staining. The transduction efficiency on human islets was significantly improved using RGD-modified adenovirus. HGF and XIAP gene expressions were dose-dependent after viral transduction. When exposed to a cocktail of inflammatory cytokines, RGD-Adv-hHGF-hXIAP-transduced human islets showed decreased caspase 3 activity and reduced apoptotic cell death. Prolonged normoglycemic control could be achieved by transplanting RGD-Adv-hHGF-hXIAP-transduced human islets. Immunofluorescence staining of kidney sections bearing RGD-Adv-hHGF-hXIAP-transduced islets was positive for insulin and von Willebrand factor (vWF) at 200 days after transplantation. These results indicated that ex vivo transduction of islets with RGD-Adv-hHGF-hXIAP decreased apoptotic islet cell death and improved islet revascularization, and eventually might improve the outcome of human islet transplantation.