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志物，用于无应答乳腺癌化学预防试验。为此，将开发替代生物标志物，由乳腺细胞（肿瘤前、早期肿瘤）分泌的蛋白组成，可在血清或乳头抽吸液中进行测定。将在大鼠逆转录病毒载体Neu-induced乳腺癌模型中鉴定和验证这些分泌的蛋白质生物标志物。将使用该模型的两种配置，其中一种是给予卵巢切除大鼠供试药物，第二种是用他莫昔芬和供试药物治疗完整大鼠。使用的药物包括：单萜紫苏醇;类视色素9-顺式视黄酸（或Targretin）和4-HPR;考克斯-2抑制剂塞来昔布;和EGFR激酶抑制剂。这些化合物中的几种已被证明可以预防所提出的大鼠模型中的卵巢非反应性癌症。将从给药和对照大鼠中采集DCIS和乳腺癌组织。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组织之间的基因表达差异。这些差异表达的基因将进行信息过滤，以产生那些（约10%）编码分泌蛋白。在使用定量PCR确认这些选定基因的差异基因表达后，将对其进行蛋白质组学分析。该分析将确定这些候选基因编码的蛋白质是否在药物治疗与对照大鼠的肿瘤前和肿瘤性乳腺细胞中差异表达，以及它们是否以药物治疗可改变的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来自患有各种良性、肿瘤前或肿瘤性乳腺疾病的患者的非应答性乳腺癌、乳腺导管液和血清中进行定量。该项目将为II期临床乳腺癌预防试验提供替代生物标志物和特异性检测，重点是对乳腺癌无反应的乳腺癌。这些基于分泌到血液或乳腺导管液中的蛋白质的替代预防生物标志物将能够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.