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，是个体易感性与终生暴露于营养性和毒性化合物相互作用的结果。癌症的干细胞理论假定，干细胞自我更新和分化途径的畸变在癌变中是不可或缺的。大多数体外癌症研究是在克隆群体扩增的癌细胞或适应培养条件的细胞系中进行的。这些研究通常没有考虑到个体间的差异或以前的暴露，并且可能在提供化学预防营养治疗的生理学相关数据方面受到限制。在这里，我们提出了一个新的体外模型来了解两种化合物的转录组效应，这两种化合物已被证明可以影响干细胞的自我更新：饮食中的多酚姜黄素和生物碱胡椒碱。我们之前使用乳腺球模型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，同时对正常分化的细胞无毒。在Specific Aim 1中，我们将在锚定独立条件下培养原代乳腺细胞和癌细胞系，并使用姜黄素和胡椒碱单独或联合治疗。我们将提取RNA进行深度测序，采用条形码分析测序法（Bar-seq）。生物信息学分析将确定姜黄素和胡椒碱在癌细胞和正常原代乳腺细胞中的作用机制的差异。在Specific Aim 1中鉴定的差异表达基因将在Specific Aim 2中扩大的原发组织和乳腺癌细胞系样本集中进行验证。我们在体外应用这种新颖的方法将为理解姜黄素和胡椒碱特异性靶向干细胞的机制以及在正常细胞和肿瘤细胞中的作用机制差异提供关键环节。此外，本研究将提供姜黄素和胡椒碱治疗效果的干细胞特异性生物标志物，这将扩展到未来的临床研究中。这些数据将为营养化合物，如姜黄素和胡椒碱，是否可以在易感人群中作为预防乳腺癌的干预措施提供基础证据。