Angiogenesis-targeting microbubbles combined with ultrasound-mediated gene therapy in brain tumors
Angiogenesis-targeting microbubbles combined with ultrasound-mediated gene therapy in brain tumors
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DOI：
10.1016/j.jconrel.2017.04.010
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发表时间：
2017-01-10
影响因子：
10.8
通讯作者：
Yeh, Chih-Kuang
中科院分区：
文献类型：
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作者：
Chang, En-Ling;Ting, Chien-Yu;Yeh, Chih-Kuang
脑癌基因治疗的主要挑战是由于血脑屏障（BBB）和传统脑内注射引起的神经损伤导致转基因表达不佳。通过系统性经血管途径使用超声靶向微泡（MB）振荡的非病毒基因递送很有吸引力，但目前还没有高产且靶向的基因表达方法。在这项研究中，我们开发了一种非病毒和血管生成靶向基因递送方法，可在不损伤脑的情况下进行有效的脑肿瘤基因治疗。我们开发了一种针对 VEGFR2 的阳离子微泡 (VCMB) 基因载体，用于与经颅聚焦超声 (FUS) 暴露一起使用，以实现瞬时基因传递。该系统在脑肿瘤模型中进行了测试，使用萤火虫荧光素酶基因和单纯疱疹病毒 1 型胸苷激酶/更昔洛韦 (pHSV-TK/GCV) 和 VCMB，在 FUS 暴露下进行转基因表达和抗肿瘤作用。体外数据表明，VCMBs 具有高 DNA 负载效率和对癌细胞的高亲和力。体内数据证实，该技术增强了基因向肿瘤组织的传递，而不影响正常脑组织。相对于 CMB 组（1.9 倍）和直接注射组，VCMB 组产生更高的荧光素酶表达（3.8 倍）。在 FUS 下使用 VCMB 进行 pHSV-TK/GCV 系统治疗的大鼠中，第 25 天的肿瘤体积 (9.7 +/- 5.2 mm(3)) 明显小于直接注射组 (40.1 +/- 4.3 mm(3))。我们展示了负载 DNA 的 VCMB 和 FUS 的成功使用，以非病毒、非侵入性和靶向基因递送至脑肿瘤。
The major challenges in gene therapy for brain cancer are poor transgene expression due to the blood-brain barrier (BBB) and neurologic damage caused by conventional intracerebral injection. Non-viral gene delivery using ultrasound-targeted microbubbles (MBs) oscillation via the systematic transvascular route is attractive, but there is currently no high-yielding and targeted gene expression method. In this study, we developed a non-viral and angiogenesis-targeting gene delivery approach for efficient brain tumor gene therapy without brain damage. We developed a VEGFR2-targeted and cationic microbubbles (VCMBs) gene vector for use with transcranial focused ultrasound (FUS) exposure to allow transient gene delivery. The system was tested in a brain tumor model using the firefly luciferase gene and herpes simplex virus type 1 thymidine kinase/ganciclovir (pHSV-TK/GCV) with VCMBs under FUS exposure for transgene expression and anti-tumor effect. In vitro data showed that VCMBs have a high DNA-loading efficiency and high affinity for cancer cells. In vivo data confirmed that this technique enhanced gene delivery into tumor tissues without affecting normal brain tissues. The VCMBs group resulted in higher luciferase expression (3.8 fold) relative to the CMBs group (1.9 fold), and the direct injection group. The tumor volume on day 25 was significantly smaller in rats treated with the pHSV-TK/GCV system using VCMBs under FUS (9.7 +/- 5.2 mm(3)) than in the direct injection group (40.1 +/- 4.3 mm(3)). We demonstrated the successful use of DNA-loaded VCMBs and FUS for non-viral, non-invasive and targeted gene delivery to brain tumors.