Gene Therapy for HPV-Associated Malignancies

描述（由申请人提供）：在本申请中，我们提出构建和测试用于治疗HPV相关宫颈癌的新型腺病毒（Ad）载体，其将直接溶瘤作用与诱导针对HPV阳性肿瘤细胞的强细胞免疫应答结合联合收割机。本文所述的条件复制型Ad载体（Ad.IR）将仅在肿瘤细胞中表达热休克蛋白（HSP）gp 96和HPV-16 E7蛋白。我们将使用具有源自E7+鼠肿瘤细胞的肝转移的宫颈癌的免疫活性小鼠模型来测试以下：E7和HSP都具有增加Ad载体的溶瘤活性的能力，并且我们假设我们的载体将由于这些蛋白质的肿瘤特异性表达而有效地杀死肿瘤细胞。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）应答的靶点的潜力，我们假设gp 96的表面表达和E7的过表达将刺激有效的抗E7和抗肿瘤CTL应答。C.我们假设，免疫处理和呈现的坏死物质释放的肿瘤细胞裂解后，广告载体介导的溶瘤将增强CTL反应，这些活动将相互补充。最后，即使用病毒载体转导肿瘤的速率有限，抗肿瘤免疫应答也会导致最初未转导的肿瘤细胞裂解。

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.