Microbubble-mediated delivery of human adenoviruses does not elicit innate and adaptive immunity response in an immunocompetent mouse model of prostate cancer
Microbubble-mediated delivery of human adenoviruses does not elicit innate and adaptive immunity response in an immunocompetent mouse model of prostate cancer
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DOI：
10.1186/s12967-019-1771-0
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发表时间：
2019-01-11
影响因子：
7.4
通讯作者：
Howard, Candace M.
中科院分区：
文献类型：
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作者：
De Carlo, Flavia;Thomas, Litty;Howard, Candace M.
背景利用人腺病毒(HADS)将基因转移到恶性部位，由于其免疫原性和宿主特异性，一直受到限制。小鼠细胞通常缺乏HADS附着所需的某些受体，因此小鼠细胞一般不允许人腺病毒感染和复制，这限制了翻译研究。方法我们开发了一种基因转移方法，该方法使用脂质包裹的全氟化碳微泡和超声波相结合的方法来保护和传递HADS到靶组织，而不需要特定的受体。结果在体外模型中，我们发现小鼠TRAMP-C2和人DU145前列腺癌细胞在参与HADS黏附和内化的受体的表达模式上是相似的。HAD-GFP(绿色荧光蛋白)转导细胞的百分率在24小时后呈剂量依赖性增加，48小时和72小时后GFP基因得到有效表达。为了评估我们的图像引导递送系统是否能够在体内有效地保护HAD免受免疫系统的攻击，我们将HAD-GFP/MB复合体注射到健康免疫活性小鼠(C57BL/6)或同基因前列腺癌小鼠(TRAMP-C2)中。值得注意的是，我们没有观察到先天(TNF-和IL-6细胞因子)或获得性免疫反应(中和抗体，INF-+CD8(+)T细胞)的激活。结论本研究使我们更接近于证明建立小鼠癌症模型的可行性，以研究超声靶向微泡破坏介导的图像引导的部位特异性腺病毒基因治疗的临床翻译。
BackgroundGene transfer to malignant sites using human adenoviruses (hAds) has been limited because of their immunogenic nature and host specificity. Murine cells often lack some of the receptors needed for hAds attachment, thus murine cells are generally non-permissive for human adenoviral infection and replication, which limits translational studies.MethodsWe have developed a gene transfer method that uses a combination of lipid-encapsulated perfluorocarbon microbubbles and ultrasound to protect and deliver hAds to a target tissue, bypassing the requirement of specific receptors.ResultsIn an in vitro model, we showed that murine TRAMP-C2 and human DU145 prostate cancer cells display a comparable expression pattern of receptors involved in hAds adhesion and internalization. We also demonstrated that murine and human cells showed a dose-dependent increase in the percentage of cells transduced by hAd-GFP (green fluorescent protein) after 24h and that GFP transgene was efficiently expressed at 48 and 72h post-transduction. To assess if our image-guided delivery system could effectively protect the hAds from the immune system in vivo, we injected healthy immunocompetent mice (C57BL/6) or mice bearing a syngeneic prostate tumor (TRAMP-C2) with hAd-GFP/MB complexes. Notably, we did not observe activation of innate (TNF- and IL-6 cytokines), or adaptive immune response (neutralizing antibodies, INF-+ CD8(+) T cells).ConclusionsThis study brings us a step closer to demonstrating the feasibility of murine cancer models to investigate the clinical translation of image guided site-specific adenoviral gene therapy mediated by ultrasound-targeted microbubble destruction.