Adeno-associated viral vectors penetrate human solid tumor tissue in vivo more effectively than adenoviral vectors
Adeno-associated viral vectors penetrate human solid tumor tissue in vivo more effectively than adenoviral vectors
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DOI：
10.1089/104303402753812511
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发表时间：
2002-06-01
影响因子：
4.2
通讯作者：
Hoover, F
中科院分区：
文献类型：
--
作者：
Enger, PO;Thorsen, F;Hoover, F
研究了腺相关病毒载体（AAV，血清2型）和腺病毒载体（ADV，血清5型）在三种不同癌症模型中的转导效率。首先，我们使用流式细胞术定量分析了来自恶性组织（6例胶质瘤、6例乳腺癌和1例白血病）的13种细胞系中AAV-GFP或ADV-GFP的转导。这些实验表明，不同细胞系之间的转导效率不同（0%-81%），在13个细胞系中，有9个细胞系的ADV比AAV更有效。其次，用表达GFP或lacZ磁带的ADV或AAV感染人胶质母细胞瘤制备的球体，2周后，在球体上观察到一致的报告基因表达。尽管AAV在整个球体中产生一致的转导，但ADV感染主要局限于球体的外层细胞层，这表明AAV在穿透实体肿瘤组织时更有效。第三，将多形性胶质母细胞瘤患者的人体活检组织移植到裸鼠体内，培养4周后注射病毒载体。结合使用高分辨率磁共振成像（MRI）和组织学分析，可以识别转导细胞及其在肿瘤内的空间分布。aav介导的转基因表达可以在整个肿瘤的细胞簇中观察到，而av介导的转导仅限于肿瘤周围的细胞。因此，虽然AAV和ADV载体对肿瘤来源细胞系的感染程度相似，但与ADV载体相比，AAV对胶质母细胞瘤球体和异种移植物的渗透效率更高。这些结果提示AAV可能适合用于恶性肿瘤的治疗性基因传递。
The transduction efficiencies of adeno-associated viral vectors (AAV, serotype 2) and adenovirus vectors (ADV, serotype 5) were examined in three different models of cancer. First, we used flow cytometry to quantitate AAV-GFP or ADV-GFP transduction in 13 cell lines derived from malignant tissue (6 gliomas, 6 mammary cancers, and 1 leukemia). These experiments showed variable transduction efficiency (0%-81%) between the cell lines, with ADV being more effective compared to AAV in 9 of 13 cell lines. Second, spheroids prepared from human glioblastomas were infected with ADV or AAV expressing GFP or lacZ cassettes, and after 2 weeks, uniform reporter gene expression was observed on the spheroid. Whereas AAV produced consistent transduction throughout the spheroids, ADV infection was mainly limited to the outer cell layers of the spheroids, suggesting that AAV were more efficient at penetrating solid tumor tissue. Third, human biopsies from glioblastoma multiforme patients were xenografted into nude rats and grown for 4 weeks followed by viral vector injection. Combined use of high-resolution magnetic resonance imaging (MRI) and histologic analysis allowed the identification of transduced cells and their spatial distribution within the tumors. AAV-mediated transgene expression was observed in cell clusters through the entire tumor, while ADV-mediated transduction was restricted to cells at the tumor periphery. Thus, while AAV and ADV vectors may infect tumor-derived cell lines to a similar degree, AAV penetrated glioblastoma spheroids and xenografts more efficiently compared to ADV vectors. These results suggest that AAV may be suitable for therapeutic gene delivery to malignant tumors.