The role of Twist in inducing breast cancer initiating cells and metastasis

Twist 在诱导乳腺癌起始细胞和转移中的作用

• 批准号: 8039739
• 负责人: Venu Raman
• 金额: $ 23.39万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-14 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8039739
• 关键词: ABCC1 gene; Autopsy; BHLH Protein; Benign; Biogenesis; Biological Markers; Biological Process; Breast; Breast Cancer Cell; Breast Carcinoma; CD44 gene; Cancer cell line; Cataloging; Catalogs; Cell Cycle Regulation; Cell Line; Cells; Choline; Choline Kinase; Clinical; Development; Epithelial; Epithelial Cells; Epithelium; Exhibits; Fatty acid glycerol esters; Gene Expression; Genetic; Goals; Growth; Human; Image; Implant; In Vitro; Knowledge; Lead; Link; Longitudinal Studies; Loss of E-cadherin Expression; MCF7 cell; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Mesenchymal; Metabolic; Metastatic Lesion; Modeling; Molecular; Mucin 1 protein; Myoepithelial; Neoplasm Metastasis; Neoplastic Cell Transformation; Noninfiltrating Intraductal Carcinoma; Oncogenic; Organ; Outcome; Pathogenesis; Pathway interactions; Patients; Permeability; Pharmaceutical Preparations; Phenotype; Population; Pre-Clinical Model; Property; Protein p53; Regulation; Research; Research Design; Research Project Grants; Research Proposals; Rhodamine 123; Role; SCID Mice; Sampling; Sorting - Cell Movement; Staging; Stem cells; Technology; Testing; Time; Tissue Sample; Transcriptional Regulation; Treatment outcome; Tumor Cell Invasion; Visceral; Work; Xenograft procedure; advanced disease; aldehyde dehydrogenases; angiogenesis; base; breast tumorigenesis; cancer cell; clinical application; improved; in vivo; insight; malignant breast neoplasm; metabolomics; novel; optical imaging; progenitor; programs; radiation resistance; research study; therapy resistant; tumor; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): The overall objective of this research proposal is to delineate the biological functions of Twist in promoting cancer initiating cells and metastasis in the pathogenesis of breast cancer. In support for this objective, we had earlier demonstrated that over-expression of Twist, a basic helix-loop-helix transcription factor, in breast cells (MCF-7) promotes epithelial-mesenchymal transition, enhanced tumorigenesis, increased genetic instability, and increased vasculature and permeability within the tumor microenvironment of orthotopic xenografts. We also demonstrated that the over-expression of Twist in breast cells can promote breast cancer initiating cell phenotype characterized by high CD44 expression, little or no CD24 expression and increased ALDH activity. In addition, Twist over-expressing cells exhibit high efflux of Hoechst 33342 and Rhodamine 123 as a result of increased expression of ABCC1 (MRP1) transporters, a property of cancer initiating cells. Importantly, we show that inoculums of only twenty cells of the Twist over-expressing CD44?/low sub-population are capable of forming tumors in the mammary fat pad of SCID mice. In our recent studies, a direct association between Twist expression and the dysregulation of Mucin 1 (MUC1) was demonstrated in breast cancer cells. Interestingly, our studies have also revealed that Twist expression regulates the expression of p27 and choline kinase, which are well-established markers of aggressive cancer phenotypes. In this revised application, we aim to establish the mechanistic basis for the role of Twist in pathogenesis of breast cancer and in altering metabolomics during the ontogeny of breast cancer. Our hypothesis is that Twist expression alters normal cellular pathways, facilitating the differentiation towards myoepithelial progenitor cells and increasing metastatic potential. To test this hypothesis, we propose three specific aims. In Aim 1, we will identify cancer initiating cells induced by Twist and functionally demonstrate their tumor forming potential. In Aim 2, we will determine the molecular mechanisms of Twist induced breast cancer progression and metastasis through its regulation of p27 and choline kinase. In addition, we will longitudinally determine the metabolic changes induced by Twist expression in preclinical models. In Aim 3, we will identify early markers of aggressive phenotypes and cancer initiating markers induced by Twist expression in pure DCIS samples and utilize the samples from the rapid autopsy program to determine if Twist expression can be associated with organ specific breast cancer metastasis. Importantly, we will identify breast cancer initiating cells in patient breast tumors and determine their functional implications in breast tumorigenesis. Taken together these studies will establish the role of Twist in inducing breast cancer initiating cells and metastasis as well as its utility as an early marker of breast cancer. PUBLIC HEALTH RELEVANCE: The project will help define the role of Twist in inducing cancer initiating cells and in metastasis during the pathogenesis of breast cancer. The clinical translatability of these findings will be confirmed in human breast cancer tissue samples such as pure DCIS and that obtained from the rapid autopsy program at Johns Hopkins. We anticipate that the clinical applications derived from the insights and knowledge provided by the successful completion of this research project will lead to molecular approaches to identify and target this lethal subpopulation of cells, which will improve treatment outcomes. In the long term, these studies will help characterize breast cancer initiating cells and provide insights that will contribute to the development of novel imaging markers and chemotherapeutic drugs for breast cancer.

描述（由申请人提供）：本研究提案的总体目标是描述Twist在乳腺癌发病机制中促进癌症起始细胞和转移的生物学功能。为了支持这一目标，我们先前已经证明，在乳腺细胞（MCF-7）中过表达Twist（一种碱性螺旋-环-螺旋转录因子）可促进上皮-间充质转化，增强肿瘤发生，增加遗传不稳定性，并增加原位异种移植物肿瘤微环境中的血管和渗透性。我们还证明了乳腺细胞中Twist的过表达可以促进乳腺癌起始细胞表型，其特征在于高CD 44表达，很少或没有CD 24表达和增加的ALDH活性。此外，Twist过表达细胞表现出Hoechst 33342和罗丹明123的高外排，这是由于ABCC 1（MRP 1）转运蛋白表达增加所致，这是癌症起始细胞的一种特性。重要的是，我们表明，只有20个细胞的Twist过表达CD 44？/低亚群能够在SCID小鼠的乳腺脂肪垫中形成肿瘤。在我们最近的研究中，在乳腺癌细胞中证明了Twist表达与粘蛋白1（MUC 1）失调之间的直接关联。有趣的是，我们的研究还揭示了Twist表达调节p27和胆碱激酶的表达，这是侵袭性癌症表型的公认标志物。在这个修订后的应用程序中，我们的目标是建立机制的基础上，扭曲的作用，乳腺癌的发病机制，并在改变代谢组学在乳腺癌的个体发生。我们的假设是Twist表达改变了正常的细胞通路，促进了向肌上皮祖细胞的分化并增加了转移潜力。为了验证这一假设，我们提出了三个具体目标。在目标1中，我们将鉴定由Twist诱导的癌症起始细胞，并在功能上证明它们的肿瘤形成潜力。目的二：探讨Twist对p27和胆碱激酶的调节作用及其诱导乳腺癌进展和转移的分子机制。此外，我们将在临床前模型中纵向确定Twist表达诱导的代谢变化。在目标3中，我们将鉴定纯DCIS样品中由Twist表达诱导的侵袭性表型的早期标志物和癌症起始标志物，并利用来自快速尸检程序的样品来确定Twist表达是否与器官特异性乳腺癌转移相关。重要的是，我们将在患者乳腺肿瘤中识别乳腺癌起始细胞，并确定其在乳腺肿瘤发生中的功能意义。总之，这些研究将确立Twist在诱导乳腺癌起始细胞和转移中的作用，以及其作为乳腺癌早期标志物的实用性。 公共卫生相关性：该项目将有助于确定Twist在乳腺癌发病过程中诱导癌症起始细胞和转移中的作用。这些发现的临床可转化性将在人类乳腺癌组织样本中得到证实，例如纯DCIS和从约翰霍普金斯的快速尸检项目中获得的样本。我们预计，从成功完成该研究项目所提供的见解和知识中获得的临床应用将导致分子方法来识别和靶向这种致命的细胞亚群，这将改善治疗结果。从长远来看，这些研究将有助于表征乳腺癌起始细胞，并提供有助于开发新型乳腺癌成像标记物和化疗药物的见解。