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 建议通过开发干扰异常的试剂来实现这一点 人肺癌和发育不良的支气管上皮细胞中的信号通路 细胞。我们将通过经气管滴注的方式给这些药物 雾化吸入在无菌大鼠临床前模型中的应用 正交异性人类肺癌。我们提出了一种循序渐进的方法 首先是对人肺癌和支气管上皮的体外研究 细胞系。这些研究之后是正常的动物研究，以 评价毒性、给药和药代动力学参数及原位试验 评估肿瘤传递和反应的动物研究。第一阶段和第二阶段 人类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，治愈方法仍然存在 低至13%，主要是因为当时癌症已经转移 诊断。因此，新的早期发现、预防和治疗战略 都是需要的。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和神经肽激活血管 这些细胞中的细胞内信号通路。这一途径由以下因素激活 多肽-受体结合。受体与异三聚体G相连 (AlphaGq家族、β和Gamma家族的蛋白质)，当激活时 转而激活磷脂酶Cβ(PLCβ)。激活的PLCβ裂解 与受体结合的三磷酸肌醇(IP3)的膜脂 内质网导致细胞内钙升高。这个 PLCβ的激活也会导致蛋白激酶C的激活 甘油二酯。游离钙激活胞质蛋白， 随后作为转录因子，导致细胞增殖。 在过去的孢子授予期间，我们支持G多肽的激活 蛋白质也可以导致Mekk途径的激活。更改 结构性活性GTPase缺陷Gq蛋白的信号通路 通过逆转录病毒或通过P物质治疗引入的基因 衍生品导致了不和谐的信号。不和谐的信号是 其特点是抑制PLCβ激活，抑制钙离子 通过刺激MEKK释放和抑制增殖 细胞凋亡的途径和诱导。我们还展示了一系列新的 二聚缓激肽拮抗剂也抑制钙释放和抑制 在诱导细胞凋亡的同时促进生长。我们开始探索这些产品的交付 经气管滴注和气雾剂的遗传和药理作用 在裸鼠和大鼠原位模型中吸入。我们还展示了 新型PKC/钙调蛋白抑制剂地塞米松抑制人小细胞肺癌细胞生长 体外和体内，通过完成第二阶段研究的 I期研究，并已将14名患者聚集到I期研究。在.期间 未来5年，我们将完成这项第二阶段的地塞米松研究；完成 G蛋白基因及其反义寡聚体的体外评价 聚阳离子脂类和人类逆转录病毒载体；优化体内递送 聚阳离子中反义寡核苷酸和突变细胞信号基因的研究 脂类和逆转录病毒载体在荷瘤裸鼠体内的实验研究 产后气管内滴注和雾化吸入； 并测定最有效的新物质P物质和缓激肽 对抗者。我们认为这些新的治疗和预防 策略将在未来5年应用于人类受试者，并将 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.