Transcriptomic effects of curcumin and piperine in breast stem cells

姜黄素和胡椒碱对乳腺干细胞的转录组作用

• 批准号: 8304701
• 负责人: LAURA ROZEK
• 金额: $ 7.78万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8304701
• 关键词: Accounting; Address; Affect; Agonist; Alkaloids; Area; Bioinformatics; Biological; Biological Assay; Biological Markers; Breast; Breast Cancer Cell; Breast Cancer Prevention; Cancer cell line; Cell Extracts; Cell Line; Cell model; Cells; Chemopreventive Agent; Clinical; Clinical Research; Clonal Expansion; Complex; Curcumin; Data; Dietary Factors; Dietary Polyphenol; Disease; Estrogen Receptors; Estrogen receptor negative; Estrogen receptor positive; Exposure to; Future; Gene Expression Profile; Genes; Genetic Predisposition to Disease; Genetic Transcription; Genome; In Vitro; Individual; Intervention; Link; Malignant Neoplasms; Mammary Gland Parenchyma; Measures; Methodology; Methods; Modeling; Nutrient; Nutritional; Pathway interactions; Patients; Poison; Population; Predisposition; Prevention strategy; Preventive; Property; RNA; RNA Sequences; Research; Risk Reduction; Sampling; Sequence Analysis; Stem cells; Therapeutic; Tissue Sample; Tissues; Treatment Efficacy; Work; cancer cell; cancer prevention; carcinogenesis; effective therapy; genome-wide; in vitro Model; malignant breast neoplasm; member; neoplastic cell; next generation; novel; piperine; preclinical study; response; self-renewal; stem; stem cell population; theories; toxicant; transcriptomics; treatment effect

DESCRIPTION (provided by applicant): Sporadic cancer is a complex disease, a consequence of the interaction of individual susceptibility with a lifetime of exposures to both nutritive and toxic compounds. The stem cell theory of cancer postulates that aberrations in stem cell self renewal and differentiation pathways are integral in carcinogenesis. Most in vitro cancer studies are undertaken in cancer cells expanded from a clonal population or in cell lines that have adapted to culture conditions. These studies often do not account for interindividual variability or previous exposures and may be limited in their ability to produce physiologically relevant data about chemopreventive nutrient treatment. Here, we propose a novel in vitro model to understand the transcriptomic effects of treatment of two compounds that have been shown to affect stem cell self renewal: the dietary polyphenol curcumin and the alkaloid piperine. We have previously used the mammosphere model to show that both curcumin and piperine specifically target breast stem cells and synergistically inhibit self-renewal while remaining non-toxic to normal differentiated cells. In Specific Aim 1, we will culture primary breast cells as well as cancer cell lines in anchorage independent conditions, and treat with curcumin and piperine, both individually and in combination. We will extract RNA for deep sequencing using the barcode analysis by sequencing method (Bar-seq). Bioinformatic analyses will indentify differences in mechanism of action of curcumin and piperine in cancer cells and normal primary breast cells. Differentially expressed genes identified in Specific Aim 1 will be validated in an expanded primary tissue and breast cancer cell line sample set in Specific Aim 2. Our application of this novel in vitro methodology will provide a crucial link in understanding the mechanism by which curcumin and piperine specifically target stem cells as well as differences in mechanisms of action in normal vs. tumor cells. Additionally, this study wil provide stem cell specific biomarkers of efficacy of treatment for curcumin and piperine that will be extended to future clinical studies. These data will provide foundational evidence as to whether nutritional compounds, such as curcumin and piperine, can be used as an intervention in breast cancer prevention in susceptible populations. PUBLIC HEALTH RELEVANCE: Sporadic breast cancer is a disease that involves the interaction between individual susceptibility and a lifetime of exposure to toxicants and nutritive compounds. In this study, we propose to develop a new in vitro model to understand how nutrients have cancer preventive properties in normal breast stem cells. Results from this study could identify methods to tell if nutritive compounds are effective treatments in future clinical studies and identify areas of the genome that are important in cancer prevention

描述（由申请人提供）：散发性癌症是一种复杂的疾病，这是个人易感性与对营养和有毒化合物的一生相互作用的相互作用。癌症的干细胞理论假设干细胞自我更新和分化途径中的畸变在癌变中不可或缺。大多数体外癌症研究都是在从克隆人群或适应培养条件的细胞系扩展的癌细胞中进行的。这些研究通常不会说明个体的变异性或以前的暴露，并且可能会受到产生有关化学预防养分治疗的生理相关数据的能力。在这里，我们提出了一个新型的体外模型，以了解两种化合物的治疗的转录组作用，这些化合物已显示出影响干细胞自我更新：饮食中的多酚姜黄素和生物碱管碱。我们先前已经使用乳球模型来表明姜黄素和胡椒碱都专门针对乳腺干细胞，并协同抑制自我更新，同时对正常分化细胞保持无毒。在特定的目标1中，我们将在独立条件下培养原代乳腺细胞以及癌细胞系，并单独和组合用姜黄素和管碱治疗。我们将通过测序方法（BAR-SEQ）提取使用条形码分析来提取RNA进行深层测序。生物信息学分析将降低癌细胞和正常原代乳腺细胞中姜黄素和果皮作用机理的差异。在特定目标1中鉴定出的差异表达的基因将在特定目标2中设置的扩展的原代组织和乳腺癌细胞系样本中进行验证。我们对这种新颖的体外方法的应用将在理解姜黄素和哌啶的机制方面提供至关重要的联系。特异性靶向干细胞以及正常和肿瘤细胞中作用机理的差异。此外，这项研究将提供干细胞特异性生物标志物的姜黄素和胡椒碱治疗功效的生物标志物，并将扩展到将来的临床研究。这些数据将提供基本的证据，表明营养化合物（例如姜黄素和胡椒碱）是否可以用作易感人群预防乳腺癌的干预措施。 公共卫生相关性：零星乳腺癌是一种疾病，涉及个人敏感性与暴露于毒物和营养的寿命之间的相互作用 化合物。在这项研究中，我们建议开发一种新的体外模型，以了解养分如何在正常乳腺干细胞中具有癌症预防特性。这项研究的结果可以识别方法来判断营养化合物在未来的临床研究中是否是有效的治疗方法，并确定基因组中对预防癌症很重要的领域