Nuclear Snail-1 in the progression of Estrogen Receptor(-) invasive breast cancer

核Snail-1在雌激素受体(-)浸润性乳腺癌进展中的作用

• 批准号: 8111466
• 负责人: ROBIN Elizabeth BACHELDER
• 金额: $ 20.49万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8111466
• 关键词: Address; Antibodies; Area; Aspirate substance; Atypia; Automobile Driving; Behavior; Biological Markers; Biology; Breast; Breast Cancer Cell; Breast Diseases; Cancer cell line; Development; Diagnosis; Ductal; Early Diagnosis; Epithelial; Estrogen Receptors; Estrogen receptor negative; Family member; Foundations; Frequencies; Future; Genes; Genetic Transcription; Growth; High Risk Woman; Human; Hyperplasia; Immunohistochemistry; In Vitro; Laboratory Finding; Lesion; Malignant - descriptor; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Messenger RNA; Molecular; Noninfiltrating Intraductal Carcinoma; Nuclear; Patients; Pilot Projects; Post-Translational Protein Processing; Premalignant; Prevention; Prospective Studies; Proteins; Protocols documentation; Regulation; Relative (related person); Risk; Signal Transduction; Snails; Specimen; Testing; Transcription Repressor/Corepressor; Tumor Cell Invasion; Validation; Woman; breast lesion; cancer initiation; cohort; high risk; malignant breast neoplasm; neoplastic cell; novel; outcome forecast; protein expression; research study; translational study; tumor; tumor growth

DESCRIPTION (provided by applicant): Currently we lack biomarkers to predict development of Estrogen Receptor-negative [ER(-)] invasive breast cancer. Progress in this area is hindered by our lack of understanding of the biology of ER(-) breast cancer initiation/progression. This application studies the regulation of ER(-) breast cancer invasion by a novel signaling network involving Snail-1, a transcriptional repressor silenced by ER. In Aim 1, we will investigate the regulation of ER(-) breast tumor cell invasion by a novel signaling network involving two transcriptional repressors (Snail-1 and ZEB1). Snail-1 and ZEB1 promote invasive tumor behavior by suppressing transcription of epithelial genes. Snail-1 and ZEB1 are frequently co-expressed in breast cancer cell lines, but it is unknown how they cooperate to promote invasive behavior. MicroRNA200 (miRNA200) family members suppress breast tumor cell invasion, and their expression levels are reduced in invasive relative to non-invasive breast tumor cells. However, the factors that initially suppress miRNA200 in breast tumor cells, thus driving ZEB1 expression and invasive behavior, are unknown. We will investigate the hypothesis that Snail-1 promotes ER(-) breast tumor cell invasion by regulating the miRNA200/ZEB1 axis. To validate our in vitro findings, we will investigate expression of the nuclear Snail-1/miRNA200/nuclear ZEB1 network in primary human ER(-) breast cancers, and retrospectively determine if expression of this network predicts patient prognosis. Snail-1 mRNA levels do not reflect Snail-1 activity because: 1) Snail-1 activity is dependent on its nuclear localization, and 2) Snail-1 localization is regulated by Snail-1 post-translational modifications. Previous studies of Snail-1 protein expression in human cancers used poorly-characterized antibodies that in some cases react with other Snail-1 family members. We have optimized an immunohistochemistry protocol for detecting nuclear Snail-1 protein using a highly-specific Snail-1 antibody. Our preliminary studies indicate that nuclear Snail-1 is expressed frequently in ER(-) but not in ER(+) ductal breast cancers. In Aim 2, we will study expression of this network in pre-malignant breast lesions obtained from a unique cohort of women at high risk for developing breast cancer. We will test if expression of this network in these pre-malignant lesions retrospectively predicts subsequent development of invasive ER(-) breast cancer. The results from this two-year study will allow us to perform power calculations necessary for a future prospective study investigating the value of a nuclear Snail-1/miRNA200/nuclear ZEB1 network as a biomarker in pre-malignant lesions that predicts development of invasive ER(-) breast cancer. Collectively, the studies in this two-year proposal will define molecular determinants of ER(-) breast tumor cell invasive behavior as a means of identifying: 1) biomarkers for early detection of invasive breast cancer, and 2) targets for prevention and treatment of invasive ER(-) breast disease. PUBLIC HEALTH RELEVANCE: Only a fraction of women diagnosed with pre-malignant breast lesions develop invasive breast tumors. Currently we lack biomarkers that predict a high risk for invasive tumor growth. Identification and validation of such a biomarker will require: 1) molecular studies of key regulators of invasion, 2) studies determining the frequency of expression of these invasion markers in pre-malignant and malignant lesions, and 3) prospective studies determining if expression of these markers in pre-malignant lesions predicts subsequent development of invasive cancer. The experiments in this two year proposal will: 1) investigate the regulation of Estrogen Receptor-negative [ER(-)] breast cancer invasion by a novel signaling network involving the transcriptional repressor Snail-1, 2) examine the frequency of expression of determinants of this Snail-1 network in malignant and pre-malignant breast lesions, and 3) retrospectively examine if expression of this Snail-1 network in pre-malignant breast lesions predicts development of invasive ER(-) breast cancer. Using the results obtained, we will perform a power calculation necessary for planning a future prospective study that tests if this signaling network is a valuable biomarker for predicting development of invasive ER(-) breast cancer. Collectively, these studies will define molecular determinants of ER(-) breast tumor cell invasive behavior as a means of identifying: 1) biomarkers for early detection of invasive ER(-) breast cancer, and 2) targets for prevention and treatment of invasive ER(-) breast disease.

描述（由申请人提供）：目前我们缺乏预测雌激素受体阴性[ER(-)]浸润性乳腺癌发展的生物标志物。 由于我们对 ER(-) 乳腺癌发生/进展的生物学缺乏了解，阻碍了这一领域的进展。 该应用研究了一种涉及 Snail-1 的新型信号网络对 ER(-) 乳腺癌侵袭的调节，Snail-1 是一种被 ER 沉默的转录抑制因子。 在目标 1 中，我们将研究涉及两种转录抑制因子（Snail-1 和 ZEB1）的新型信号网络对 ER(-) 乳腺肿瘤细胞侵袭的调节。 Snail-1 和 ZEB1 通过抑制上皮基因的转录来促进侵袭性肿瘤行为。 Snail-1 和 ZEB1 经常在乳腺癌细胞系中共表达，但尚不清楚它们如何合作促进侵袭行为。 MicroRNA200（miRNA200）家族成员抑制乳腺肿瘤细胞侵袭，其表达水平在侵袭性乳腺肿瘤细胞中相对于非侵袭性乳腺肿瘤细胞降低。 然而，最初抑制乳腺肿瘤细胞中 miRNA200 从而驱动 ZEB1 表达和侵袭行为的因素尚不清楚。 我们将研究 Snail-1 通过调节 miRNA200/ZEB1 轴促进 ER(-) 乳腺肿瘤细胞侵袭的假设。 为了验证我们的体外研究结果，我们将研究原发性人类 ER(-) 乳腺癌中核 Snail-1/miRNA200/核 ZEB1 网络的表达，并回顾性确定该网络的表达是否可以预测患者预后。 Snail-1 mRNA 水平不反映 Snail-1 活性，因为：1) Snail-1 活性取决于其核定位，2) Snail-1 定位受 Snail-1 翻译后修饰调节。 先前对人类癌症中 Snail-1 蛋白表达的研究使用了特征较差的抗体，这些抗体在某些情况下会与其他 Snail-1 家族成员发生反应。 我们优化了使用高度特异性 Snail-1 抗体检测核 Snail-1 蛋白的免疫组织化学方案。 我们的初步研究表明，核 Snail-1 在 ER(-) 导管癌中频繁表达，但在 ER(+) 导管癌中不表达。 在目标 2 中，我们将研究该网络在癌前乳腺病变中的表达，这些病变取自一组独特的乳腺癌高危女性群体。 我们将测试该网络在这些癌前病变中的表达是否能够回顾性预测侵袭性 ER(-) 乳腺癌的后续发展。 这项为期两年的研究结果将使我们能够进行未来前瞻性研究所需的功效计算，以调查核 Snail-1/miRNA200/核 ZEB1 网络作为癌前病变生物标志物的价值，预测侵袭性 ER 的发展（-） 乳腺癌。 总的来说，这项为期两年的提案中的研究将定义 ER(-) 乳腺肿瘤细胞侵袭行为的分子决定因素，作为识别：1）早期检测侵袭性乳腺癌的生物标志物，以及 2）预防和治疗乳腺癌的目标。侵袭性 ER(-) 乳腺疾病。 公共卫生相关性：只有一小部分被诊断患有癌前乳腺病变的女性会发展为侵袭性乳腺肿瘤。 目前，我们缺乏预测侵袭性肿瘤生长高风险的生物标志物。 这种生物标志物的鉴定和验证将需要：1）对侵袭的关键调节因子进行分子研究，2）确定这些侵袭标志物在癌前和恶性病变中的表达频率的研究，以及3）确定这些侵袭标志物的表达是否存在的前瞻性研究癌前病变中的标记物可预测随后发展为浸润性癌症。 这个为期两年的提案中的实验将：1) 研究涉及转录抑制因子 Snail-1 的新型信号网络对雌激素受体阴性 [ER(-)] 乳腺癌侵袭的调节，2) 检查决定簇的表达频率Snail-1 网络在恶性和癌前乳腺病变中的表达，以及 3) 回顾性检查该 Snail-1 网络在癌前乳腺病变中的表达是否预测侵袭性 ER(-) 乳腺的发展 癌症。 利用获得的结果，我们将进行必要的功效计算，以规划未来的前瞻性研究，测试该信号网络是否是预测侵袭性 ER(-) 乳腺癌发展的有价值的生物标志物。 总的来说，这些研究将定义 ER(-) 乳腺肿瘤细胞侵袭行为的分子决定因素，作为识别以下内容的一种手段：1) 用于早期检测侵袭性 ER(-) 乳腺癌的生物标志物，2) 预防和治疗侵袭性 ER 的靶点(-) 乳腺疾病。