Targeting pERK1/2 in Human Mammary Cancer Stem Cells

靶向人类乳腺癌干细胞中的 pERK1/2

基本信息

批准号：
8509637
负责人：
SHEHLA PERVIN
金额：
$ 26.51万
依托单位：
CHARLES R. DREW UNIVERSITY OF MED & SCI
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：
2011-07-11 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8509637
关键词：
African African American Aggressive behavior Attenuated Behavior Biological Assay Biology Breast Cancer Cell CD44 gene Cancer Patient Cancer cell line Caucasians Caucasoid Race Cell Line Cells Clinical Complex Data Diagnosis Disease Drug Design ERBB2 gene Effectiveness Estrogen receptor negative Event Fluorescence-Activated Cell Sorting Fresh Tissue Gelatinase B Genes Goals HRAS gene Human Immunohistochemistry In Vitro MAP Kinase Gene MAPK3 gene MMP9 gene Malignant - descriptor Mammary Neoplasms Mammary gland Mediating Mesenchymal Metastatic Neoplasm to the Breast Molecular N-Cadherin Nuclear Nuclear Pore Complex Proteins Nude Mice Oncogenic Outcome Oxidative Stress PECAM1 gene Paraffin Embedding Pathway interactions Patients Pharmaceutical Preparations Play Population Postmenopause Premenopause Property Reporting Role Signal Transduction Small Interfering RNA Snails Sorting - Cell Movement Stem cells Testing Therapeutic Time Tissue Sample Tumor Biology Tumor Volume Undifferentiated Vascular Endothelial Growth Factors Vimentin Western Blotting Woman Xenograft procedure aldehyde dehydrogenase 1 angiogenesis base cancer stem cell cell motility cell type chemotherapy clinical practice design embryonic stem cell epithelial to mesenchymal transition health disparity in vivo mRNA Expression malignant breast neoplasm matrigel mortality new therapeutic target progesterone receptor negative protein expression self-renewal slug small hairpin RNA triple-negative invasive breast carcinoma tumor tumor initiation tumor progression tumorigenic

项目摘要

DESCRIPTION (provided by applicant): Breast cancer is a complex disease associated with specific morphological and clinical features. Significant health disparity and high mortality rate is reported in African American (AA) patients, who suffer from unique and highly aggressive breast tumors. AA triple negative (TN) breast cancer patients suffer worse outcomes to chemotherapy compared with Caucasian women. Aggressive TN breast tumors contain poorly differentiated cells and express embryonic stem cell specific gene sets. These poorly differentiated mammary cancer stem cells (MCSCs) are the most tumorigenic cell types that drive initiation and progression of breast cancers. RAS/Raf/ERK1/2 signaling cascade has been found to promote every aspect of breast tumor progression including aggressive behavior like high angiogenesis and motility. We hypothesize that "Mammary cancer stem cells drive aggressive TN breast tumors in AA women through sustained ERK1/2 signaling." We will test this hypothesis with the following Specific Aims: 1) Establish whether mammary cancer stem cells from AA TN breast tumors form aggressive xenografts in nude mice. 2) Examine whether sustained ERK1/2 increases aggressive behavior of mammary cancer stem cells enriched mammospheres from AA TN breast tumors. 3) Examine whether inhibition of ERK1/2 signaling is effective in attenuating aggressive properties of mammary cancer stem cells from AA TN breast tumors. Fluorescence-activated cell sorting (FACS) will be used to enrich MCSCs (Lin-/CD44+/CD24- /ALDH1+) from TN AA and Caucasian breast tumors, which will injected into the nude mice to form xenotransplants. These xenografts will be used to compare tumor volume, expression of angiogenesis stimulating factors (VEGF, CD31, MMP9), and motility of cells (Boyden Chamber Assay) between the two groups. MCSCs will also be enriched as mammospheres and pERK1/2-mediated effect on cell motility and expression of angiogenesis stimulating factors will be analyzed. ERK1/2 signaling will be attenuated in MCSCs by Nup153 shRNA and its effect on tumor promoting behavior will be analyzed both in vitro and in vivo. Increased understanding of the critical role of ERK1/2 signaling in MCSCs from AA patients and may provide novel targets for therapeutic drug design.

描述（由申请人提供）：乳腺癌是一种与特定形态和临床特征相关的复杂疾病。据报道，非裔美国人（AA）患者有明显的健康差异和高死亡率，这些患者患有独特且高度侵略性的乳腺肿瘤。与高加索妇女相比，AA三重阴性（TN）乳腺癌患者对化学疗法的预后较差。侵袭性TN乳腺肿瘤包含分化较差的细胞和表达胚胎干细胞特异性基因集。这些分化差的乳腺癌干细胞（MCSC）是驱动乳腺癌启动和进展的最肿瘤细胞类型。发现RAS/RAF/ERK1/2信号传导级联反应可促进乳腺肿瘤进展的各个方面，包括高血管生成和运动等攻击行为。我们假设“乳腺癌干细胞通过持续的ERK1/2信号传导促进AA女性的侵袭性TN乳腺肿瘤。”我们将以以下特定目的检验该假设：1）确定来自AA TN乳腺肿瘤的乳腺癌干细胞是否在裸鼠中形成侵袭性异种移植物。 2）检查持续的ERK1/2是否增加了乳腺癌干细胞的侵略性行为，富含AA TN乳腺肿瘤的哺乳动物。 3）检查抑制ERK1/2信号传导是否有效地减弱了来自AA TN乳腺肿瘤的乳腺癌干细胞的侵袭性。荧光激活的细胞分选（FACS）将用于富含TN AA和高加索乳腺肿瘤的MCSC（LIN-/CD44+/CD24-/ALDH1+），该乳腺肿瘤将注射到裸鼠中以形成Xenotplansplants。这些异种移植物将用于比较两组之间的肿瘤体积，血管生成刺激因子的表达（VEGF，CD31，MMP9）以及两组之间细胞（Boyden室分析）的运动。 MCSC也将作为哺乳动物球富集，PERK1/2介导对细胞运动的影响和血管生成刺激因子的表达。 NUP153 SHRNA将在MCSC中衰减ERK1/2信号，其对肿瘤促进行为的影响将在体外和体内分析。对AA患者的MCSC中ERK1/2信号传导的关键作用的了解增加，并可能为治疗药物设计提供新的靶标。