Gene Therapy for HPV-Associated Malignancies

描述（由申请人提供）：在本申请中，我们建议构建并测试一种用于治疗HPV相关宫颈癌的新型腺病毒（Ad）载体，该载体将结合直接溶瘤作用和诱导针对HPV阳性肿瘤细胞的强细胞免疫反应。本文描述的条件复制Ad载体（Ad. ir）将在肿瘤细胞中特异性表达热休克蛋白（HSP） gp96和hpv - 16e7蛋白。我们将采用一种免疫能力强的小鼠宫颈癌肝转移模型来测试以下内容：a . E7和热休克蛋白都有能力增加Ad载体的溶瘤活性，我们假设我们的载体可以有效地杀死肿瘤细胞，因为这些蛋白的肿瘤特异性表达。B.基于热休克蛋白的免疫刺激特性和E7作为细胞毒性t细胞（CTL）反应靶点的潜力，我们假设gp96的表面表达和E7的过表达将刺激有效的抗E7和抗肿瘤CTL反应。C.我们假设，Ad载体介导的肿瘤溶解后肿瘤细胞释放的坏死物质的免疫处理和呈递将增强CTL反应，并且这些活动将相互补充。最后，即使病毒载体的肿瘤转导率有限，抗肿瘤免疫反应也会导致最初未转导的肿瘤细胞溶解。

DESCRIPTION (provided by applicant): In this application we propose to construct and test a novel adenovirus (Ad) vector for therapy of HPV associated cervical carcinoma that will combine a direct oncolytic effect with an induction of strong cellular immune responses against HPV-positive tumor cells. The conditionally replicating Ad vector (Ad.IR) described here will express the heat shock protein (HSP) gp96 and the HPV-16 E7 protein exclusively in tumor cells. We will employ an immunocompetent mouse model of cervical carcinoma with liver metastases derived from E7+ murine tumor cells to test the following: A. Both E7 and HSPs have an ability to increase the oncolytic activity of Ad vectors and we hypothesize that our vector will efficiently kill tumor cells as a result of tumor-specific expression of these proteins. B. Based on the immunostimulatory properties of HSPs and the potential of E7 to serve as a target for a cytotoxic T-cell (CTL) response, we hypothesize that surface expression of gp96 and overexpression of E7 will stimulate efficient anti-E7 and anti-tumor CTL responses. C. We hypothesize that the immune processing and presentation of necrotic material released from tumor cells lysed following Ad vector mediated oncolysis will augment the CTL response and that these activities will complement each other. Finally, even if the rate of tumor transduction with the viral vector is limited, an anti-tumor immune response will lead to lysis of tumor cells that were originally not transduced.