Genotype and phenotype predictors in therapy response in renal cell carcinoma

摘要 近年来，肾细胞癌（RCC）的治疗已被靶向化疗的疗效所改变。 抑制参与关键细胞信号传导途径的激酶和VEGF的药物， 肿瘤血管生成然而，患者对这些治疗剂的反应是高度可变的，并且目前 无法根据临床或病理学数据或现有的实验室/遗传检测进行预测。的 该提案的目标是为最常见的 RCC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- 抑制肿瘤血管活化，肿瘤血管的活化状态和稳定性是主要的 反应的决定因素。我们还假设，ccRCC肿瘤血管的表型与以下因素有关： 肿瘤细胞的分子病理学特别是，由于这些药物还可以抑制肿瘤细胞， 信号传导和调节它们的行为，ccRCC肿瘤细胞信号传导，HIF-1/HIF-2（缺氧诱导的 因子）表达和VHL功能是反应的潜在决定因素。检查参数 血管表型、肿瘤细胞表型和VHL基因型作为治疗反应的预测因子， ccRCC标本将接受适当生物标志物抗原的多重免疫染色， 通过新型计算机辅助图像分析系统进行分析，该系统客观地量化分析物 基于细胞（细胞计数）的染色以及通过传统的基于像素的分析。这些研究将 在接受单药贝伐珠单抗、舒尼替尼或 索拉非尼在正在进行的多机构II期（ECOG 2804）和III期（ECOG 2805）临床试验中， 使用来自90、170和170名适当ccRCC患者的亚组的肿瘤块， 各自的药物。本提案的具体目的是（目的1）分析血管和内皮细胞 ccRCC肿瘤中细胞活化表型和血管周细胞覆盖率;（目的2）分析肿瘤细胞 ccRCC肿瘤中的信号传导、HIF表型和VHL基因型;以及（目的3）相关参数 在先前的目标中量化彼此之间的关系，治疗结果和发展 使用生物统计学和生物信息学的治疗反应的简约预测模型 方法。这些研究的结果应该能够确定最合适的药物， 治疗个体ccRCC患者，并协助合理开发二线和 联合药物治疗。除了ccRCC，研究中的靶向药物也用于治疗 越来越多的癌症，以及为ccRCC开发的反应预测因子， 最好对这些药物进行研究，因为它们作为单一药物使用， 这些其他癌症对掺入靶向药剂的组合疗法的反应。

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.