INTRAPERITONEAL AD5-P53 GENE THERAPY FOR OVARIAN CANCER

申请人的描述（Applicant's Description） 卵巢癌仍然是最致命的妇科恶性肿瘤， 通常失败的外科减积和多种方案的传统 化疗 数十年来在剂量递增、药物开发和 新的递送系统在以下方面没有显示出任何显著的进步： 生存 了解卵巢癌的分子基础， 特别是肿瘤抑制基因p53的作用，激发了人们的想法， 新的治疗基因靶向策略。 体外和体外 临床前数据支持使用腺病毒p53基因治疗（Ad 5-p53） 用于治疗人实体瘤。 自从 晚期卵巢癌通常留在腹膜腔室中， 腹水中的肿瘤细胞易于用于生物学研究， 腹腔注射Ad 5-p53是一种理想的模型， 将最近的实验室发展转化为临床试验。 这 拟议的I期临床试验将评估以下假设： 腹腔注射腺病毒p53基因治疗 与卵巢癌将导致最低限度的毒性生长抑制， 通过瞬时诱导肿瘤细胞凋亡减少肿瘤。 我们将 通过以下主要具体目标实现这一目标：具体目标1： 确定腹腔注射的最大耐受剂量和毒性 给予基于腺病毒的p53基因治疗患有腹水的患者 晚期、复发性或持续性卵巢癌常规治疗失败 化疗 腹膜内施用Ad 5-p53的剂量递增 将在标准临床I期试验中累积的患者中进行研究。 器官毒性、副作用特征以及最大耐受剂量 (MTD)将被确定。 具体目的2：评估Ad 5-p53载体 包括基因转导在内的药代动力学和生物学功效， 转基因表达和肿瘤细胞杀伤机制。 肿瘤细胞 腹水将被用于研究的效果和时间过程的作用 AdS-p53。 免疫组化、PCR、RT-PCR等方法 将使用特异性引物、流式细胞术和TUNEL测定。 具体 目的3：测定人全身和腹腔免疫功能 对病毒载体的反应，全身生物分布，载体的可能性 感染性和载体灭活，腹膜内给药后 Ad 5-p53的施用。 宿主对Ad 5-p53的反应将在 通过细胞病变效应（CPE）试验测定血清和腹水上清液，补体 以及CH 50测量和宿主抗体测量。 这将决定 安全性和不太可能的传染性。 具体目标4：确定 在晚期、复发性和复发性乳腺癌患者中， 或是卵巢癌 缓解的次要终点为 通过临床或放射学肿瘤测量测量，间接 测量腹水产生和症状特征的变化。

DESCRIPTION (Applicant's Description) Ovarian cancer remains the most lethal gynecologic malignancy, as patients commonly fail surgical debulking and multiple regimens of conventional chemotherapy. Decades of research in dose escalation, drug development and novel delivery systems has failed to show any significant advance in survival. Understanding of the molecular basis of ovarian cancer, in particular the role of the tumor suppressor gene p53, has stimulated ideas on new therapeutic gene targeting strategies. In vitro and in vitro preclinical data support the use of adenoviral p53 gene therapy (Ad5-p53) for use in the treatment of human solid tumors. Since the spread of advanced ovarian cancer typically remains in the peritoneal compartment and tumor cells in ascites are easily accessible for biologic studies, intraperitoneal administration of Ad5-p53 is an ideal model to use in translating recent laboratory developments into clinical trials. This proposed phase I clinical trial will evaluate the hypothesis that intraperitoneal administration of adenoviral p53 gene therapy in patients with ovarian cancer will result in a minimally toxic growth inhibition and tumor reduction by transient induction of tumor cell apoptosis. We will accomplish this by the following primary specific aims: Specific Aim 1: To determine the maximal tolerated dose and toxicities of intraperitoneally administered adenoviral-based p53 gene therapy in patients wit ascites from advanced, recurrent or persistent ovarian carcinoma failing conventional chemotherapy. Dose escalations of intraperitoneal administration of Ad5-p53 will be studied in patients accrued in a standard clinical phase I trial. Organ toxicities, side effect profiles as well as the maximal tolerated dose (MTD) will be determined. Specific Aim 2: To evaluate the Ad5-p53 vector pharmacokinetics and biologic efficacy including gene transduction, transgene expression, and mechanism bf tumor cell kill. Tumor cells in ascites will be used to study the effect and time course of action of AdS-p53. Methods such as immunohistochemistry, PCR, RT-PCR with vector specific primers, flow cytometry and TUNEL assays will be used. Specific Aim 3: To determine the human systemic and intraperitoneal immunologic response to the viral vector, systemic biodistribution, potential for vector infectivity and vector inactivation, following intraperitoneal administration of ad5-p53. Host response to Ad5-p53 will be studied on serum and ascitic supernatant by cytopathic effect (CPE) assays, complement and CH50 measurement and host antibody measurements. This will determine the safety and unlikely infectivity profile. Specific Aim 4: To determine the response rate to the Ad-p53 vector in patients with advanced, recurrent or persistent ovarian cancer. A secondary endpoint of response will be measured by clinical or radiographic tumor measurements, indirect measurements of ascites production and changes in symptom profile.