Targeting pERK1/2 in Human Mammary Cancer Stem Cells

靶向人类乳腺癌干细胞中的 pERK1/2

• 批准号: 8677817
• 负责人: SHEHLA PERVIN
• 金额: $ 27.35万
• 依托单位: CHARLES R. DREW UNIVERSITY OF MED & SCI
• 依托单位国家: 美国
• 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/8677817
• 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; MCF10A cells; MDA MB 231; 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

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) 患者的健康状况存在显着差异，且死亡率较高，他们患有独特且高度侵袭性的乳腺肿瘤。与白人女性相比，AA 三阴性 (TN) 乳腺癌患者的化疗结果较差。侵袭性 TN 乳腺肿瘤含有分化不良的细胞并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）是驱动乳腺癌发生和进展的最具致瘤性的细胞类型。研究发现 RAS/Raf/ERK1/2 信号级联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侵袭行为。我们假设“乳腺癌干细胞通过持续的 ERK1/2 信号传导在 AA 女性中驱动侵袭性 TN 乳腺肿瘤。”我们将通过以下具体目标来检验这一假设：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 是否会增加来自 AA TN 乳腺肿瘤的富含乳腺癌干细胞的乳腺球的侵袭行为。 3) 检查抑制 ERK1/2 信号传导是否能有效减弱来自 AA TN 乳腺肿瘤的乳腺癌干细胞的侵袭特性。将使用荧光激活细胞分选（FACS）从TN AA和高加索乳腺肿瘤中富集MCSC（Lin-/CD44+/CD24-/ALDH1+），将其注射到裸鼠中形成异种移植。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（VEGF、CD31、MMP9）的表达和细胞运动性（Boyden Chamber Assay）。 MCSC 也将被富集为乳腺球，并且将分析 pERK1/2 介导的细胞运动和血管生成刺激因子表达的影响。 Nup153 shRNA 将减弱 MCSC 中的 ERK1/2 信号传导，并将在体外和体内分析其对肿瘤促进行为的影响。加深对 AA 患者 MCSC 中 ERK1/2 信号传导关键作用的了解，并可能为治疗药物设计提供新靶点。