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

基本信息

批准号：
8230596
负责人：
ROBIN Elizabeth BACHELDER
金额：
$ 17.07万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8230596
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 public health relevance research study translational study tumor tumor growth

项目摘要

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（ - ）]浸润性乳腺癌的发展。由于我们对ER（ - ）乳腺癌启动/进展的生物学缺乏了解，这一领域的进步受到了阻碍。该应用研究了通过涉及蜗牛1的新型信号网络对ER（ - ）乳腺癌侵袭的调节，蜗牛1（一种被ER沉默的转录阻遏物）调节。在AIM 1中，我们将通过一个涉及两个转录阻遏物（Snail-1和Zeb1）的新型信号网络来调查ER（ - ）乳腺肿瘤细胞侵袭。 Snail-1和Zeb1通过抑制上皮基因的转录来促进侵入性肿瘤行为。 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以促进侵入性行为。 MicroRNA200（miRNA200）家庭成员抑制乳腺肿瘤细胞的侵袭，相对于非侵入性乳腺肿瘤细胞，其侵入性降低了其表达水平。然而，最初抑制乳腺肿瘤细胞中miRNA200的因素，从而驱动Zeb1表达和侵入性行为。我们将研究蜗牛-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发现，我们将研究原代人ER（ - ）乳腺癌中核蜗牛-1/miRNA200/核Zeb1网络的表达，并回顾性地确定该网络的表达是否预测了患者的预后。蜗牛-1 mRNA水平不反映蜗牛1活性，因为：1）蜗牛1活性取决于其核定位，而2）蜗牛-1定位受到蜗牛-1翻译后修饰的调节。先前对人类癌症中蜗牛-1蛋白表达的研究使用了特征较差的抗体，在某些情况下，这些抗体与其他蜗牛-1家族成员有反应。我们优化了一种使用高度特异性的蜗牛-1抗体检测核蜗牛-1蛋白的免疫组织化学方案。我们的初步研究表明，核蜗牛1在ER（ - ）中经常表达，但在ER（+）导管性乳腺癌中不经常表达。在AIM 2中，我们将研究该网络的表达，这些乳腺癌具有高风险的独特女性乳腺癌的乳腺癌病变。我们将测试该网络在这些恶性病变中的表达是否回顾性地预测随后的侵入性ER（ - ）乳腺癌的发展。这项为期两年的研究的结果将使我们能够进行未来的前瞻性研究所必需的功率计算，以研究核蜗牛-1/miRNA200/核Zeb1网络的价值作为预防性病变中的生物标志物，以预测侵入性ER（ - ）乳腺癌的发展。总的来说，这项为期两年的提案中的研究将定义ER（ - ）乳腺肿瘤细胞侵入性行为的分子决定因素，作为识别的一种手段：1）生物标志物，用于早期发现浸润性乳腺癌的生物标志物，以及2）预防和治疗侵入性ER（ - ）乳腺病的靶标。公共卫生相关性：只有一小部分被诊断出患有恶性乳房病变的妇女会出现侵入性乳腺肿瘤。目前，我们缺乏预测侵入性肿瘤生长的高风险的生物标志物。对这种生物标志物的鉴定和验证将需要：1）对入侵关键调节剂的分子研究，2）确定这些侵袭标记物在恶性病变和恶性病变中表达的频率的研究，以及3）前瞻性研究，确定这些标志物在预防性病变中的表达是否预测侵入性癌症的随后发展。这两年的建议中的实验将：1）调查通过一个新的信号传导网络侵入雌激素受体受体阴性[ER（ - ）]涉及转录抑制剂蜗牛1的新型信号网络的侵袭侵入性ER（ - ）乳腺癌。使用获得的结果，我们将执行计划未来的前瞻性研究所必需的功率计算，该研究是否是该信号网络是否是预测侵入性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（ - ）乳腺肿瘤细胞侵入性行为的分子决定因素，作为识别的一种手段：1）用于早期检测侵入性ER（ - ）乳腺癌的生物标志物，以及2）靶标，以预防和治疗侵入性ER（ - ）乳腺病。