INHIBITION OF PEPTIDE SIGNAL TRANSDUCTION--THERAPY & PREVENTION OF LUNG CANCER

该项目的总体目标是开发新的代理商和路线 用于人类肺癌的化学预防和治疗。 我们 我建议通过开发一种干扰异常的 肺癌与支气管上皮异型增生信号通路 细胞 我们将通过经气管滴注提供这些药物， 在临床前模型中使用无胸腺大鼠进行雾化吸入， 正交异性人类肺癌。 我们提出了一个逐步的方法， 从人类肺癌和支气管上皮细胞的体外研究开始， 细胞系 这些研究之后是正常的动物研究， 评价毒性、递送和药代动力学参数， 动物研究以评估肿瘤递送和反应。 I期和II期 人体临床研究是评估过程的最后一步。 肺 癌症是美国最致命的癌症， 低至13%，主要是因为癌症已经转移， 诊断. 因此，新的早期发现、预防和治疗战略 是必要的。 许多肺癌（所有小细胞肺癌和一些非小细胞肺癌） 小细胞肺癌）具有神经内分泌表型。 这些细胞 表达多种神经肽受体，神经肽激活 这些细胞中的细胞内信号通路。 这条通路被激活， 肽-受体结合。 受体与异源三聚体G 蛋白质（alphaGq家族和β和γ），当激活时， 进而激活磷脂酶C β（PLC β）。 活化的PLC β裂解 三磷酸肌醇（IP 3），它结合到 使内质网内钙离子升高。 的 PLC β激活还导致蛋白激酶C的激活， 甘油二酯 游离钙激活细胞质蛋白， 随后作为转录因子导致细胞增殖。 在过去的SPORE资助期间，我们发现G 蛋白质也可以导致MEKK途径的激活。 改变 组成型活性GT3缺陷Gq蛋白信号通路 通过逆转录病毒或P物质处理引入的基因 衍生物导致不一致的信号传导。 不和谐的信号是 其特征在于抑制PLC β活化，抑制钙离子通道， 通过刺激MEKK释放和抑制增殖 途径和诱导凋亡。 我们还发现了一系列新的 二聚体缓激肽拮抗剂也抑制钙释放， 生长同时诱导凋亡。 我们开始探索如何将这些 通过气管滴注和气雾剂的遗传和药理学试剂 在原位裸鼠和大鼠模型中吸入。 我们还发现， 一种新的PKC/钙调蛋白抑制剂，dexniguldipine，抑制SCLC生长， 在体外和体内，通过完成一项 I期研究，并已将14名患者纳入I期研究。 期间 未来5年，我们将完成这项II期右尼古地平研究;完成 在体外评价G蛋白基因和反义寡聚体在 聚阳离子脂质和人逆转录病毒载体;优化体内递送 反义寡核苷酸和突变细胞信号基因在聚阳离子 脂质和逆转录病毒载体在荷原位肺癌裸鼠中的应用 经阴道滴注和雾化吸入给药后; 确定最有效的新P物质和缓激肽 对手。 我们相信这些新的治疗和预防方法 在未来5年内，这些策略将应用于人类受试者， 最终导致肺癌死亡率的提高。

The overall objective of this project is to develop new agents and routes of delivery for the chemoprevention and treatment of human lung cancer. We propose to do this by developing agents which interfere with aberrant signal pathways in human lung cancer and dysplastic bronchial epithelial cells. We will deliver these agents by transtracheal instillation and aerosolized inhalation in a preclinical model using athymic rats bearing orthotropic human lung cancers. We propose a stepwise approach which begins with in vitro studies in human lung cancer and bronchial epithelial cell lines. These studies are followed by normal animal studies to evaluate toxicity, delivery and pharmacokinetic parameters and orthotopic animal studies to evaluate tumor delivery and response. Phase I and II human clinical studies re athe last step in the evaluation process. Lung cancer is the most lethal cancer in the United States and the cure remains low at 13% primarily because th cancers have metastasized at the time of diagnosis. Thus, new early detection, prevention and treatment strategies are needed. Many lung cancers (all small cell lung cancers and some non- small cell lung cancers) have a neuroendocrine phenotype. These cells express multiple neuropeptide receptors and neuropeptides activate an intracellular signal pathway in these cells. This pathway is activated by peptide-receptor binding. The receptors are linked to heterotrimeric G proteins ( of the alphaGq family and beta and gamma) which when activate in turn activate phospholipase Cbeta (PLCbeta). Activated PLCbeta cleaves membrane lipids to inositol trisphosphate (IP3) which binds to receptors on the endoplasmic reticulum resulting in a rise intracellular calcium. The PLCbeta activation also results in activation of protein kinase C through diacylglycerol. The free calcium activates cytoplasmic protein which subsequently act as transcription factors leading to cell proliferation. During the past SPORE grant period, we shoed that peptide activation of G proteins can also lead to activation of the MEKK pathway. Alteration of the signal pathway by constitutively active GTPase deficient Gq protein genes introduced in a retrovirus or by treatment with substance p derivatives led to discordant signaling. The discordant signaling was characterized by inhibition of PLCbeta activation, inhibition of calcium release and inhibition of proliferation with stimulation of the MEKK pathway and induction of apoptosis. We also showed that a series of new dimeric bradykinin antagonists also inhibited calcium release and inhibited growth while inducing apoptosis. We began to explore the delivery of these genetic and pharmacologic agents via transtracheal instillation and aerosol inhalation in an orthotopic nude mouse and rat model. We also showed that a novel PKC/calmodulin inhibitor, dexniguldipine, inhibited SCLC growth in vitro and in vivo, determined a dose the phase II studies by completing a phase I study and have accrued 14 patients to a phase I study. During the next 5 years, we will complete this phase II dexniguldipine study; complete in vitro evaluation of G protein genes and antisense oligomers delivered in polycationic lipids and human retrovirus vectors; optimize in vivo delivery of antisense oligonucleotides and mutant cell signal genes in polycationic lipid and retroviral vectors in nude rats bearing orthotopic lung cancers after delivery by intratracheal instillation and aerosolized inhalation; and determine maximally effective new substance P and bradykinin antagonists. We believe that these new treatment and prevention strategies will be applied to human subjects in the next 5 years and will ultimately lead to advances in lung cancer mortality.