Genotype and phenotype predictors in therapy response in renal cell carcinoma

摘要 近年来，肾细胞癌(RCC)的治疗因靶向疗效的改变而发生变化 抑制关键细胞信号通路中的激酶和血管内皮生长因子的药物，血管内皮生长因子是 肿瘤血管生成。然而，患者对这些疗法的反应是高度不同的，目前 不能根据临床或病理数据或现有的实验室/基因检测进行预测。这个 这项建议的目标是开发治疗反应的预测因子，用于最常见的 肾细胞癌的亚型，透明细胞(CcRCC)，基于新的测试。我们的方法主要基于 假设ccRCC治疗中使用的靶向药物-贝伐单抗、舒尼替尼和索拉非尼- 抑制肿瘤血管的激活，肿瘤血管的激活状态和稳定性是主要的 反应的决定因素。我们还假设ccRCC肿瘤血管的表型与 肿瘤细胞的分子病理生物学。特别是，由于这些药物还可以抑制肿瘤细胞 信号和调节其行为，ccRCC肿瘤细胞信号，HIF-1/HIF-2(低氧诱导 因子)表达和VHL功能是反应的潜在决定因素。检查参数的步骤 血管表型、肿瘤细胞表型和VHL基因作为治疗反应的预测因子 CcRCC样本将接受适当的生物标记物抗原和 由一种新的计算机辅助图像分析系统进行分析，该系统可以客观地定量分析物 在细胞(细胞学)基础上以及传统的基于像素的分析的基础上进行染色。这些研究将 单药贝伐单抗、舒尼替尼或单药治疗慢性肾细胞癌患者的肿瘤 正在进行的多机构第二阶段(ECOG2804)和第三阶段(ECOG2805)临床试验中的索拉非尼， 使用来自90、170和170名合适的ccRCC患者子集的肿瘤块进行治疗 各自的药物。这项建议的具体目标是(目标1)分析血管和内皮 肾细胞癌的细胞活化表型和血管周细胞覆盖率；(目标2)分析肿瘤细胞 CcRCC肿瘤的信号转导、HIF表型和VHL基因分型；以及(目标3)相关参数 在之前的目标中量化，目的是相互之间的关系，治疗结果和发展 用生物统计学和生物信息学建立简约的疗效预测模型 接近了。这些研究的结果应该能够确定最合适的治疗药物 治疗单个慢性肾细胞癌患者，并协助合理开发二线和 联合用药疗法。除了ccrcc外，正在研究的靶向药物也用于治疗。 不断扩大的癌症数量，以及为ccRCC开发的反应预报器，其中 这些试剂是最好的研究对象，因为它们是作为单一试剂使用的，可能对预测有用 这些其他癌症对合并靶向药物的联合治疗的反应。

ABSTRACT Therapy of renal cell carcinoma (RCC) has been transformed in recent years by the efficacy of targeted agents that inhibit kinases involved in critical cellular signaling pathways and VEGF, a primary driver of tumor angiogenesis. However, patient response to these therapeutics is highly variable and currently cannot be predicted based on clinical or pathological data or available laboratory/genetic testing. The goal of this proposal is to develop predictors of therapeutic response for patients with the most common subtype of RCC, clear cell (ccRCC), based on novel tests. Our approach is based primarily on the hypothesis that the targeted agents used in ccRCC therapy - bevacizumab, sunitinib and sorafenib - inhibit tumor vessel activation and that the activation status and stability of tumor vessels are major determinants of response. We also hypothesize that the phenotype of ccRCC tumor vessels is linked to the molecular pathobiology of the tumor cells. In particular, as these drugs also can inhibit tumor cell signaling and modulate their behavior, ccRCC tumor cell signaling, HIF-1/HIF-2 (hypoxia-inducible factor) expression and VHL function are potential determinants of response. To examine parameters of vascular phenotype, tumor cell phenotype and VHL genotype as predictors of response to therapy, ccRCC specimens will undergo multiplex immunostaining for the appropriate biomarker antigens and be analyzed by a novel computer-assisted image analysis system that objectively quantifies analyte staining on a cellular (cytometric) basis as well as by traditional pixel-based analysis. These studies will be performed on tumors of ccRCC patients treated with single-agent bevacizumab, sunitinib or sorafenib in ongoing multi-institutional phase II (ECOG2804) and phase III (ECOG2805) clinical trials, using tumor blocks from a subset of 90, 170 and 170 appropriate ccRCC patients for therapy with the respective drugs. The specific aims of this proposal are (Aim 1) to analyze the vascular and endothelial cell activation phenotype and vessel pericyte coverage in ccRCC tumors; (Aim 2) to analyze tumor cell signaling, HIF phenotype and VHL genotype in ccRCC tumors; and (Aim 3) to correlate parameters quantified in the prior aims for relationships to each other, to therapeutic outcome and to develop parsimonious predictive models of therapeutic response using biostatistical and bioinformatics approaches. The results of these studies should allow identification of the most appropriate drugs for treating individual patients with ccRCC and assist in the rational development of second-line and combination drug therapies. Beyond ccRCC, the targeted agents under study are used in the therapy of an ever expanding number of cancers, and the response predictors developed for ccRCC, where these agents are best studied because they are used as single-agents, may be useful for predicting response of these other cancers to combination therapy incorporating the targeted agents.