Pancreatic Cancer Treatment Using Surgery & Gene Therapy

描述（由申请人提供）：胰腺癌是第五大 美国癌症死亡原因。由于没有有效的 治疗绝大多数患有这种可怕的 胰腺癌是少数几种发病率高的疾病之一， 等于死亡治疗这些患者的新方法是 迫切需要。我们已经开发出一种癌症特异性细胞毒性基因疗法， 在SCID小鼠模型中似乎是成功的人胰腺癌。 这项提议的核心令人兴奋的新发现是， 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 可以使用大鼠胰岛素启动子-胸苷激酶 构建体和脂质体基因递送系统，随后是更昔洛韦 治疗该提案的具体目的是为了测试 假设：具体目的I：确定a）PDX-1、BETA-2、加塔-4和/或 E47转录因子调控大鼠胰岛素启动子在人胰岛素细胞中的表达 胰腺癌细胞系，B）这些转录因子存在于 切除的人胰腺癌和转移瘤，和c） 大鼠胰岛素启动子-胸苷激酶和更昔洛韦体外可增强 用PDX-1，BETA 2，GATA 4， 或E47。具体目的二：确定大鼠胰岛素 启动子-胸苷激酶和更昔洛韦可用于消融人 胰腺癌细胞在SCID小鼠体内通过a）iv相对于ip脂质体相对于 腺病毒递送系统B）重复的大鼠胰岛素启动子胸苷激酶 基因递送和延长的更昔洛韦治疗，c）递增剂量的大鼠 胰岛素启动子胸苷激酶和d）确定细胞毒性 大鼠胰岛素启动子-胸苷激酶和更昔洛韦体内作用 体外或体内共转染增强人胰腺癌细胞 具有PDX-1、BETA2、GATA 4或E47的细胞系。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治疗将影响 使用脂质体抗腺病毒基因转染分离的小鼠和人胰岛 递送系统和B）体内脂质体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、胰岛素分泌和葡萄糖调节 在SCID小鼠中。该提案的目的是：（1）获得更大的 了解参与大鼠胰岛素激活的机制 2）获得胰腺癌细胞内启动子所需的临床前数据， 一项将联合收割机手术和系统性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-胸苷激酶是否会 在体内和体外均影响小鼠和人类胰岛。我们希望这个策略 最终将为胰腺癌患者提供更有效的治疗方法。 使用手术切除原发癌和全身胰腺癌的癌症 癌症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.