Selectivity of TAG-72-targeted adenovirus gene transfer to primary ovarian carcinoma cells versus autologous mesothelial cells in vitro.
Selectivity of TAG-72-targeted adenovirus gene transfer to primary ovarian carcinoma cells versus autologous mesothelial cells in vitro.
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发表时间：
2000-11
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通讯作者：
F. Kelly;Ryan Miller;D. Buchsbaum;J. Gómez-Navarro;M. Barnes;R. Alvarez;D. Curiel;General Surgery;Center;G-N D T C
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作者：
F. Kelly;Ryan Miller;D. Buchsbaum;J. Gómez-Navarro;M. Barnes;R. Alvarez;D. Curiel;General Surgery;Center;G-N D T C
重组腺病毒 (Ad) 载体的高效基因转移取决于靶细胞上 CAR 和 α(v) 整合素的表达。因为 Ad 还可能感染附近表达这些受体的非靶细胞，例如腹腔注射后的腹膜间皮细胞。注射后，我们假设将 Ad 基因递送至大多数卵巢癌细胞上过表达的受体（例如 TAG-72），在这种情况下使用时将增强 Ad 基因转移的选择性。一种单克隆抗体（即 CC49）已在临床上研究用于卵巢癌的免疫治疗和免疫检测，用于与中和性抗旋钮单克隆抗体的 Fab 片段构建双特异性缀合物，以通过 TAG-72 靶向 Ad 结合。这种缀合物促进了 TAG-72 特异性、不依赖于 CAR 的 Ad 报告基因转移到卵巢癌细胞系和从恶性腹水培养的原发性卵巢癌细胞中。 Fab-CC49 对肿瘤细胞具有很强的选择性，相对于非靶向 Ad，将 Ad 基因转移到原发性卵巢癌细胞的能力提高了 2 至 28 倍，同时也将基因转移到自体培养的间皮细胞的能力减少了 4 到 9 倍。这些数据表明，通过 TAG-72 靶向 Ad 可以将卵巢肿瘤腹膜内 Ad 基因转移的选择性提高 8 至 252 倍。矢量注入。这些结果还确定了合理设计靶向Ad载体以实现最终临床实用性时对候选靶受体的要求，该载体可以选择性地感染肿瘤细胞并在腹膜内注射时不伤害正常细胞。注射。这种载体可以增加基因转移并降低Ad载体的毒性，这将提高临床试验中卵巢癌细胞毒性基因治疗的治疗指数。
Efficient gene transfer by recombinant adenovirus (Ad) vectors depends on expression of CAR and alpha(v) integrin on target cells. Because Ad may also infect nearby nontarget cells expressing these receptors, such as peritoneal mesothelial cells after i.p. injection, we hypothesized that targeting Ad gene delivery to a receptor overexpressed on most ovarian carcinoma cells, such as TAG-72, would enhance the selectivity of Ad gene transfer when used in this context. A monoclonal antibody that has been investigated clinically for immunotherapy and immunodetection of ovarian carcinomas, namely CC49, was used to construct a bispecific conjugate with the Fab fragment of a neutralizing anti-knob mAb to target Ad binding via TAG-72. This conjugate facilitated TAG-72-specific, CAR-independent Ad reporter gene transfer to both ovarian cancer cell lines and primary ovarian cancer cells cultured from malignant ascites fluid. Fab-CC49 was very selective for tumor cells, augmenting Ad gene transfer to primary ovarian cancer cells 2- to 28-fold relative to untargeted Ad, while also decreasing gene transfer to autologous cultured mesothelial cells 4- to 9-fold. These data suggest that targeting Ad via TAG-72 may improve the selectivity of Ad gene transfer for ovarian tumors 8- to 252-fold on i.p. vector injection. These results also define the requirements for a candidate target receptor in the rational design of a targeted Ad vector for ultimate clinical utility, one that selectively infects tumor cells and spares normal cells on i.p. injection. Such a vector may increase gene transfer and decrease the toxicity of Ad vectors, which would improve the therapeutic index of cytotoxic gene therapy for ovarian cancer in clinical trials.