Secreted Proteins as Breast Cancer Prevention Markers

描述(由申请人提供) 该项目的目标是开发易于翻译的替代生物标记物，用于激素无反应的乳腺癌化学预防试验。为此，将开发替代生物标记物，由乳腺细胞(肿瘤前期、早期肿瘤)分泌的蛋白组成，可在血清或乳房乳头吸出液中进行检测。这些分泌蛋白生物标记物将在逆转录病毒载体Neu诱导的大鼠乳腺癌模型中得到识别和验证。这个模型将使用两种配置，一种是去卵巢的大鼠给予试验药物，另一种是完整的大鼠给予他莫昔芬和试验药物(S)治疗。使用的药物包括：单萜紫苏醇；维甲酸，9-顺式维甲酸(或塔格列汀)和4-羟丙酸；环氧合酶-2抑制剂塞来昔布；以及EGFR激酶抑制剂。在所提出的大鼠模型中，这些化合物中的几个已经被证明可以预防激素不敏感的癌症。DCIS和癌乳腺组织将从药物治疗组和对照组大鼠中收集。这些组织之间的基因表达差异将使用大鼠基因表达DNA芯片阵列和严格的统计分析来确定。这些差异表达的基因将被信息过滤，以产生那些(约10%)编码分泌蛋白的基因。在使用定量-聚合酶链式反应确认这些选定基因的差异基因表达后，将对它们进行蛋白质组学分析。这项分析将确定这些候选基因编码蛋白在药物治疗组和对照组大鼠的癌前和肿瘤乳腺细胞中是否存在差异表达，以及它们是否以可通过药物治疗而改变的数量分泌到血液或乳腺导管液中。接下来，这些大鼠分泌蛋白的人类同源物将在激素反应不敏感的乳腺癌、乳腺导管液和各种良性、癌前病变或肿瘤性乳腺疾病患者的血清中进行定量。该项目将为第二阶段临床乳腺癌预防试验提供替代生物标记物和特定的分析方法，这些试验的重点是激素反应不敏感的乳腺癌。这些代孕预防生物标记物基于分泌到血液或乳腺导管液中的蛋白质，将能够很容易地转化为临床试验，因为可以以微创方式收集用于分析的患者样本。

DESCRIPTION (provided by applicant) The goal of this project is to develop readily translatable surrogate biomarkers for hormonalty non-responsive breast cancer chemoprevention trials. For this purpose, surrogate biomarkers will be developed, consisting of breast cell (pre-neoplastic, early neoplastic) secreted proteins that can be assayed for in sera or breast nipple aspirate fluids. These secreted protein biomarkers will be identified and validated in a rat retroviral vector Neu-induced mammary cancer model. Two configurations of this model will be used including one in which ovariectomized rats are given test drugs and the second in which intact rats are treated with tamoxifen and the test drug(s). Drugs to be used include: the monoterpene Perillyl alcohol; the retinoids 9-cis retinoic acid (or Targretin) and 4-HPR; the COX-2 inhibitor Celecoxib; and an EGFR kinase inhibitor. Several of these compounds have been shown to prevent hormonally non-responsive cancer in the rat model proposed. DCIS and carcinoma mammary tissues will be collected from drug-treated and control rats. Differences in gene expression between these tissues will be determined using rat gene expression DNA-chip arrays and rigorous statistical analysis. These differentially expressed genes will be informatically filtered to yield those (approximately 10%) that encode for secreted proteins. Following confirmation of differential gene expression for these selected genes using quantitative-PCR they will be subjected to a proteomic analysis. This analysis will determine if these candidate gene-encoded proteins are differentially expressed in preneoplastic and neoplastic mammary cells of drug treated versus control rats, and whether they are secreted into the blood or mammary ductal fluids in amounts that are modifiable by the drug treatment. Next the human homologues of these rat secreted proteins will be quantified in hormonalty non- responsive breast cancers, breast ductal fluids and sera from patients with various benign, preneoplastic, or neoplastic breast diseases. This project will provide surrogate biomarkers and specific assays for phase II clinical breast cancer prevention trials focused on hormonally non-responsive breast cancer. These surrogate prevention biomarkers, based on proteins that are secreted into blood or breast ductal fluids, will be able to be readily translated into clinical trials in that patient samples for analysis can be collected in a minimally invasive manner.