Molecular & Genetic Determinants of Outcome in Breast Ca

描述（由申请人提供）：乳腺癌仍然是一个主要的公共卫生问题，在美国每年有超过190，000例新发病例。这些病例中约有1/3为局限于乳腺和腋窝淋巴结疾病的妇女。在这些患者中，积极的多模态治疗可以治愈超过50%的病例。然而，目前的预后标志物无法区分那些将被治愈的治疗，那些谁是治愈没有积极的治疗和那些注定要复发，无论治疗。为了提高生存率和适当的靶向治疗，显然需要更好的标记物和对复发生物学的更好理解。这项拟议的研究旨在通过检查宿主遗传特征对治疗结果的作用，以及检查这些宿主因素与肿瘤分子特征之间的联系，扩大我们对乳腺癌复发和预后的了解。本研究中感兴趣的宿主遗传因子是白细胞介素-6（IL-6），一种参与宿主对癌症免疫应答的多效性细胞因子。高血清IL-6水平是乳腺癌患者的特征，并且水平升高与侵袭性疾病相关，包括更多的转移部位和对标准疗法的抗性。IL-6基因启动子区的遗传多态性赋予细胞因子转录速率的功能意义。我们建议检查这些多态性单独或组合对淋巴结阳性乳腺癌患者的无病生存期（DFS）和总生存期（OS）的影响。我们建议在东部肿瘤协作组（ECOG）进行的大型多机构临床试验的背景下研究这个问题。这项研究是一项随机试验，在540名至少有10个阳性淋巴结的妇女中比较了标准蒽环类药物辅助化疗与高剂量化疗联合自体干细胞拯救，将提供生殖系DNA（来自储存的干细胞）、肿瘤标本和实质性随访（5.7年），以解决四个主要假设：（1）IL-6启动子多态性与淋巴结阳性乳腺癌患者的DFS和OS相关，（2）这种关系被肿瘤的雌激素受体状态和患者的绝经状态改变（3）这些多态性与介导临床结果的肿瘤受体和/或信号传导的特异性改变相关，和（4）这些多态性与所给予的治疗的毒性有关。我们将利用基于PCR的基因分型技术来鉴定IL-6启动子中的以下多态性：-174G/C、-597G/A、-572G/C和-373AnTn。我们将利用免疫组织化学染色来鉴定肿瘤受体（IL-6 R、gp 130、Her 2/neu、EGFR）、细胞凋亡（通过TUNEL、STAT-1、STAT-3、bcl-2）、细胞周期控制（通过MIB-1、AKT、p27）和环氧合酶-2酶活性（考克斯-2）的改变，所有这些都已被证明在体外与IL-6相互作用。Kaplan-Meier和考克斯比例风险技术将用于研究关注的相关性。这项研究将提供关于宿主环境和肿瘤行为的遗传决定因素之间相互作用的重要信息，并将提供急需的机制数据，这些数据对于利用这些信息开发这种毁灭性疾病的靶向治疗非常重要。

DESCRIPTION (provided by applicant): Breast cancer remains a major public health problem, with over 190,000 new cases in the U.S. annually. Approximately 1/3 of these cases comprise women with disease limited to the breast and axillary lymph nodes. In these patients, aggressive multi-modality treatment can be curative in just over 50% of cases. However, current prognostic markers are unable to discriminate between those who will be cured with therapy, those who are cured without aggressive therapy and those who are destined to recur regardless of therapy. In order to improve survival and appropriately target therapies, better markers and improved understanding of the biology of recurrence is clearly needed. The proposed study seeks to expand our knowledge of breast cancer recurrence and prognosis by examining the role of host genetic characteristics on treatment outcome, and examining links between these host factors and tumor molecular characteristics. The host genetic factor of interest in this study is Interleukin-6 (IL-6), a pleiotropic cytokine involved in the host immune response to cancer. High serum levels of IL-6 are characteristic in breast cancer patients, and increased level is associated with aggressive disease, including more metastatic sites and resistance to standard therapies. Genetic polymorphisms in the promoter region of the IL-6 gene confer functional significance in transcription rates of the cytokine. We propose to examine the impact of these polymorphisms, individually and in combination, on disease-free survival (DFS) and overall survival (OS) in node-positive breast cancer patients. We propose to study this issue in the context of a large, multi-institution clinical trial conducted by the Eastern Cooperative Oncology Group (ECOG). This study, a randomized trial comparing standard anthracycline-based adjuvant chemotherapy to high dose chemotherapy with autologous stem cell rescue in 540 women with at least 10 positive lymph nodes, will provide germ-line DNA (from stored stem cells), tumor specimens and substantial follow-up (5.7 years) to address four main hypotheses: (1) that IL-6 promoter polymorphisms are associated with DFS and OS in patients with node-positive breast cancer, (2) that this relationship is modified by estrogen receptor status of the tumor and menopausal status of the patient (3) that these polymorphisms are associated with specific alterations in tumor receptors and/or signaling that mediate the clinical outcomes and (4) that these polymorphisms are related to the toxicity of the treatments administered. We will utilize PCR-based genotyping techniques to identify the following polymorphism in the IL-6 promoter: -174G/C, -597G/A, -572G/C and -373AnTn. We will utilize immunohistochemical staining to identify alterations in tumor receptors (IL-6R, gp130, Her2/neu, EGFR), apoptosis (via TUNEL, STAT-1, STAT-3, bcl-2), cell cycle control (via MIB-1, AKT, p27) and cyclooxygenase-2 enzyme activity (COX-2), all of which have been shown to interact with IL-6 in vitro. Kaplan-Meier and Cox proportional hazards techniques will be used to investigate the associations of interest. This study will provide important information on interactions between genetic determinants of the host environment and tumor behavior, and will provide much-needed mechanistic data that will be important in utilizing this information for the development of targeted therapies for this devastating disease.