Pancreatic Cancer Treatment Using Surgery & Gene Therapy

简介(申请人提供)：胰腺癌居第五位 导致美国癌症死亡的原因。由于没有有效的 对绝大多数患有这种可怕疾病的患者的治疗 胰腺癌是为数不多的发病率较高的疾病之一。 等于死亡。治疗这些患者的一种新方法是 急需之物。我们已经开发出一种癌症特异性细胞毒基因疗法 人类胰腺癌在SCID小鼠模型中似乎是成功的。 这一提议的核心是令人兴奋的新发现，即大鼠胰岛素 启动子在胰腺癌中被激活，从而允许肿瘤特异性 自杀基因的激活。我们的假设是人类胰腺癌 使用大鼠胰岛素启动子-胸苷激酶可以选择性地消融细胞 更昔洛韦脂质体基因递送系统的构建及应用 治疗。该提案的具体目标旨在测试 假设：具体目标一：确定a)PDX-1、β-2、GATA-4和/或 E47转录因子调控大鼠胰岛素启动子在人体内的表达 胰腺癌细胞株，b)这些转录因子存在于 切除的人胰腺癌和转移，以及c)细胞毒作用 大鼠胰岛素启动子-胸苷激酶和更昔洛韦的体外增强作用 PDX-1、β_2、GATA_4、GATA_4共转染人胰腺癌细胞系的实验研究 或者E47。特定目的II：确定大鼠胰岛素是否 启动子-胸苷激酶和更昔洛韦可用于消融人 胰腺癌细胞通过a)iv与ip脂质体在SCID小鼠体内的比较 腺病毒递送系统b)重复的大鼠胰岛素启动子胸苷激酶 基因传递和更昔洛韦的长期治疗，c)大鼠剂量增加 胰岛素启动子胸苷激酶和d)测定是否有细胞毒作用 大鼠胰岛素启动子-胸苷激酶和更昔洛韦的体内效应 体内外共转染人胰腺癌细胞的增强作用 带有PDX-1、Beta2、GATA4或E47的线路。具体目标三：确定是否 A)大鼠胰岛素启动子-胸苷激酶和更昔洛韦的治疗将影响 分离的小鼠和人胰岛脂质体对腺病毒基因的诱导作用 给药系统和b)体内给药与腺病毒大鼠胰岛素 启动子-胸苷激酶和更昔洛韦将影响天然小鼠胰岛和/或 移植人胰岛形态、胰岛素分泌及血糖调节 在SCID小鼠身上。这项建议的目的是1)获得更大的 对大鼠胰岛素激活机制的认识 胰腺癌细胞内的启动子2)获得所需的临床前数据 外科手术与系统性RIP-TK脂质体基因结合的临床试验 治疗，以及3)确定大鼠胰岛素启动子-胸苷激酶是否 影响小鼠和人类的胰岛，包括活体和体内。我们希望这一战略 最终将为胰腺癌患者带来更有效的治疗方法 手术切除原发癌和全身性胰腺癌 肿瘤特异性基因治疗，以消除微转移。

DESCRIPTION (provided by applicant): Pancreatic cancer is the fifth leading cause of cancer death in the United States. Since there are no effective treatments for the overwhelming majority of patients with this terrible disease, pancreatic cancer is one of the few diseases where the incidence equals mortality. A new approach for the treatment of these patients is urgently needed. We have developed a cancer-specific cytotoxic gene therapy for human pancreatic cancer that appears to be successful in a SCID mouse model. The exciting new finding central to this proposal is that the rat insulin promoter is activated in pancreatic cancer, thus permitting tumor-specific activation of suicide genes. Our hypothesis is that human pancreatic cancer cells can be selectively ablated using a rat insulin promoter-thymidine kinase construct and a liposomal gene delivery system followed by ganciclovir treatment. The specific aims of the proposal have been designed to test the hypothesis: Specific Aim I: todetermine whether a) PDX-1, BETA-2, GATA-4 and/or E47 transcription factors regulate expression of rat insulin promoter in human pancreatic cancer cell lines, b) these transcription factors are present in resected human pancreatic cancer and metastases, and c) the cytotoxic effect of rat insulin promoter-thymidine kinase and ganciclovir invitrocan be enhanced by cotransfection of human pancreatic cancer cell lines with PDX-1, BETA2, GATA4, or E47. Specific Aim II: to determine whether the rat insulin promoter-thymidine kinase and ganciclovir can be used to ablate human pancreatic cancer cells invivoin SCID mice via a) iv versus ip liposomal versus adenoviral delivery systems b) repeated rat insulin promoter thymidine kinase gene delivery and prolonged ganciclovir treatment, c) escalating doses of rat insulin promoter thymidine kinase and d) to determine whether the cytotoxic effect of rat insulin promoter-thymidine kinase and ganciclovir invivocan be enhanced by invitroor invivocotransfection of human pancreatic cancer cell lines with PDX-1, BETA2, GATA4, or E47. Specific Aim III: to determine whether a)rat insulin promoter-thymidine kinase and ganciclovir treatment will affect isolated mouse and human islets invitrousing liposomal versus adenoviral gene delivery systems and b) invivoliposomal versus adenoviral rat insulin promoter-thymidine kinase and ganciclovir will affect native mouse islet and/or transplanted human islet morphology, insulin secretion and glucose regulation in SCID mice. The purposes of this proposal are to 1) acquire a greater understanding of the mechanisms involved in the activation of the rat insulin promoter within pancreas cancer cells 2) obtain the preclinical data needed for a clinical trial which will combine surgery and systemic RIP-TK liposomal gene therapy, and 3) to determine whether rat insulin promoter-thymidine kinase will affect mouse and human islets both invitroand invivo. We hope this strategy will ultimately lead to a more effective treatment for patients with pancreatic cancer using surgery to resect the primary cancer and systemic pancreatic cancer-specific gene therapy to ablate the micrometastases.