Prevention of Mammary Cancer in Her-2neu Transgenic Mice

描述（由申请人提供）：抗癌药物作用机制的详细知识是设计针对其功能的临床试验的要求。在我们过去3年的工作中，我们专注于了解四硫代钼酸盐（一种铜降低剂）如何抑制肿瘤生长和血管生成。我们一开始就推测，由于铜参与了几种血管生成因子的分泌和功能，因此铜缺乏将具有相当全面的、可能是普遍的抑制肿瘤血管生成的作用。我们在乳腺癌、前列腺癌、肺癌和鳞状细胞癌方面的工作为支持铜缺乏的全球性和普遍性影响提供了重要证据。具体来说，我们以前的工作在体外和体内乳腺癌模型的资助下，强烈暗示抑制NF κ B激活铜缺乏症作为一个关键的致病事件。武装与最近爆炸的知识从许多实验室的NF κ B系统的各种组件的功能，在这里，我们建议详细定义铜缺乏如何抑制NF κ B在癌细胞和正常和肿瘤相关的内皮细胞的激活。我们的目的是将铜缺乏对NF κ B的关键活化剂（如IL-1）分泌的抑制作用与NF κ B活化和随后的靶基因（TNF、IL-6、IL-8、IAP、MMP）合成的内在抑制分开。我们总体的经验性假设是，铜缺乏通过改变转录因子定位于细胞核后的转录激活来影响NF κ B的激活。NF κ B的活化过程包括许多步骤，从组分的合成到DNA结合和转录活化，其中Cu可以发挥作用。我们还假设，由于铜缺乏引起的生长停滞在巨大的肿瘤，抑制NF κ B活性也延伸到肿瘤细胞侵袭的关键介质，如膜1型基质金属蛋白酶（MT 1-MMPs）。在这个修订后的申请中，此外，我们考虑了另一种假设，即铜缺乏对AP-1和SP-1转录因子的影响，可能是由ERK信号的改变介导的。为了验证这些机制假说并继续详细描述铜缺乏的作用，我们提出了以下具体目标：1）了解在乳腺癌和其他癌症的体外模型中铜缺乏干扰TNF α和IL-1诱导的NF κ B活化的信号传导水平以及铜缺乏对AP-1和SP-1转录的影响。2)描述NF κ B活化和肿瘤条件内皮细胞中微管生长之间的相互作用。A）研究肿瘤细胞在暴露于肿瘤调节环境的内皮细胞中引发原始血管生长的过程的铜依赖性; B）将铜缺乏环境中IL-1分泌减少的作用与NF κ B B活化的潜在内在抑制分开。3)在Her 2/neu转基因小鼠（高NF κ B活性）和wnt转基因小鼠（正常NF κ B活性）中研究铜缺乏对早期肿瘤和导管增生和早期肿瘤病变影响的分子替代物。确定肿瘤病变、内皮和基质成分中铜缺乏作用的关键介质表达的变化。了解TM是否调节her 2和wnt转基因肿瘤中uPAR表达和erk信号传导。

DESCRIPTION (provided by applicant): Detailed knowledge of the mechanism of action of anti-cancer drugs is a requirement for the design of clinical trials tailored to their function. In our work conducted over the past 3 years, we have focused on understanding how tetrathiomolybdate, a copper lowering agent, inhibits tumor growth and angiogenesis. We surmised at the outset that since copper is involved in the secretion and function of several angiogenic factors, copper deficiency would have a fairly global, possibly general effect of inhibiting angiogenesis in tumors. Our work in breast cancer, prostate cancer, lung cancer, and squamous cell cancer provide important evidence in support of the global and generalizable effects of copper deficiency. Specifically, our previous work under this grant on in vitro and in vivo models of breast cancer has strongly implicated inhibition of NFkappaB activation by copper deficiency as a key causative event. Armed with a recent explosion of knowledge from many laboratories on the function of various components of the NFkappaB system, here we propose to define in detail how copper deficiency inhibits NFkappaB activation in cancer cells and in normal and tumor associated endothelial cells. We aim to separate the inhibitory effects of copper deficiency on the secretion of key activators of NFkappaB, such as IL-1 from the intrinsic inhibition of NFkappaB activation and subsequent synthesis if target genes (TNF, IL-6, IL-8, lAP, MMPs). Our overarching, empirical hypothesis is that copper deficiency affects NFkappaB activation by altering the activation of transcription after the factors are localized to the nucleus. There are many steps in the process of activation of NFkappaB, from synthesis of the components to DNA binding and transcription activation where Cu could play a role. We also postulate that due to the growth arrest elicited by copper deficiency in bulky tumors, the inhibition of NFkappaB activity also extends to key mediators of tumor cell invasion, such as membrane type 1 matrix metalloproteases (MT1-MMPs). In this revised application, in addition, we consider the alternative hypotheses that Cu deficiency has effects on the AP-1 and SP-1 transcription factors, possibly mediated by alterations of erk signaling. In order to test these mechanistic hypotheses and continue to delineate the action of copper deficiency in detail, we propose the following specific aims: 1) Understand the signaling level at which copper deficiency interferes with TNFalpha and IL-1- induced NFkappaB activation in in vitro models of breast and other cancers and the effects of Cu deficiency on AP-1 and SP-1 transcription. 2) Delineate the interaction between NFkappaB activation and microtubular outgrowth in tumor conditioned endothelial cells. A) Investigate the copper dependence of the process whereby tumor cells elicit outgrowth of primordial vessels in endothelial cells exposed to a tumor-conditioned milieu; B) Separate the effects of decreased IL-1 secretion in a copper poor environment from the potential intrinsic inhibition of NFkappaB activation by copper deficiency. 3) Investigate the molecular surrogates of copper deficiency effects in incipient tumor and ductal hyperplastic and early tumor lesions in Her2/neu transgenic mice (high NFkappaB activity), wnt transgenic mice (normal NFkappaB activity). Define changes in expression of key mediators of the action of copper deficiency in the tumor lesions, the endothelium, and the stromal components. Understand whether TM regulates uPAR expression and erk signaling in her2 and wnt transgenic tumors.