Immunologic aspects of targeted therapy of erbB tumors

我们的总体假设是，使用去癌蛋白单抗和干扰素-γ的顺序治疗会导致 单抗本身无法实现肿瘤细胞的表型变化。单抗诱导的表型变化敏感 细胞或允许干扰素-γ的意外行为直接作用于肿瘤。这项提案的总体目标是 是为了更深入地了解erbB介导的肿瘤发生过程，并开发出更强大的 治疗药物，用于治疗和防止出现耐药肿瘤的形成和扩散。我们将聘用 在番茄标记的乳房中表达MMTV-neu蛋白的转基因小鼠 这些细胞随机地发展成乳腺癌，就像人类乳腺癌一样。这些独特的 转基因将受到针对p185癌蛋白的靶向治疗的影响，随后是或 同时给予干扰素-γ治疗。肿瘤的出现和扩散将被跟踪，细胞系将从 进展性肿瘤用于进一步的生化研究。其他研究将采用配对注射抗p185的方法。 单抗和干扰素-γ，以及一种新的具有修饰的效应结构域的抗erbB-ScFv的使用，其中 我们从基因上连接了干扰素-γ分子。MMTV Neut-tdTomato转基因小鼠将成为 观察单抗单独或联合干扰素-γ对肿瘤进展和转移扩散的影响。 对使用抗p185neu单抗治疗后出现的耐药肿瘤进行表型和等位基因研究。 通过使用繁殖的细胞系，特别是我们拥有的基因，在肿瘤本身发生的变化 被认为与恶性表型有关。一种新型人源化单抗的研究 MAb-干扰素-γ重组蛋白与人源化ZZ结构域(我们称之为ZED结构域)融合将是 扩展到对赫赛汀耐药的乳腺癌。这种高度工程化的单抗物种同时使 P185erbB2/neu激酶；通过Zed结构域放大免疫球蛋白效应；最后，提供 干扰素-γ直接作用于人肿瘤细胞。我们将评估靶向抗erbB2单抗的效应域是否也被修饰 影响由erbB2/neu癌基因和其他体细胞突变(如kiras)转化的细胞。这 构建可能是一种治疗人类疾病的原型。最后，我们还将评估是否 细胞毒性/遗传毒性信号(如紫杉烷、多西紫杉醇)与这种新的抗体进一步结合使用 消除对赫赛汀有抗药性的肿瘤细胞。

Our overall hypothesis is that ordered therapy with oncoprotein disabling mAb followed by IFN-γ leads to phenotypic changes in tumor cells never achieved with mAb itself. MAb induced phenotypic changes sensitize cells or are permissive for unexpected actions of IFN-γ directly on the tumor. The overall goal of this proposal is to gain a deeper understanding of events of erbB mediated tumorigenesis and to develop more potent therapeutics to treat and prevent the emergence of resistant tumor formation and spread. We will employ MMTV NeuT-tdTomato transgenic mice created to express MMTV-neu proteins in Tomato tagged mammary cells and which develop breast tumors stochastically much like human breast cancer. These unique transgenics will be subjected to the effects of targeted therapy against the p185 oncoprotein followed by or concurrent with IFN-γ therapy. Tumor emergence and spread will be followed and cell lines developed from progressive tumors for further biochemical studies. Other studies will employ paired administration of anti-p185 mAb and IFN-γ, as well as use of a new species of anti-erbB2-scFv with a modified effector domain to which we have genetically attached the IFN-γ molecule. The MMTV NeuT-tdTomato transgenic mice will be hosts to follow the effects of mAb alone, or in combination with IFN-γ, on tumor progression and metastatic spread. Resistant tumors that arise despite treatment with mAb to p185neu will be studied for phenotypic and allelic changes that occur in the tumor itself, through the use of propogated cell lines, in particular genes we have considered to be relevant to the malignant phenotype. Our studies on a newly engineered mAb humanized mAb-IFN-γ recombinant protein fused with a humanized ZZ domain, (which we term the ZED domain) will be extended to Herceptin resistant breast cancers. This highly engineered mAb species simultaneously disables p185erbB2/neu kinases; amplifies immunoglobulin effects through the ZED domain; and, finally, delivers IFN-γ directly to human tumor cells. We will evaluate if effector domain modified targeting anti-erbB2 mAb also affects cells transformed by both the erbB2/neu oncogene and other somatic mutations such as KiRas. This construct may be a prototype of a therapeutic for human disease. Finally, we will also evaluate if cytotoxic/genotoxic signals (such as the taxane, docetaxol) used with this new species of antibody further eliminates Herceptin resistant tumor cells.