Cholangiocyte microRNAs Regulate Mcl-1 and Cell Death

胆管细胞 microRNA 调节 Mcl-1 和细胞死亡

• 批准号: 8128448
• 负责人: Justin L. Mott
• 金额: $ 13.23万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-20 至 2011-09-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8128448
• 关键词: 3' Untranslated Regions; Address; Apoptosis; Apoptosis Regulator; Area; Basic Science; Bile duct carcinoma; Binding; Biological Assay; Biology; Cancer Biology; Cell Culture Techniques; Cell Death; Cell model; Cells; Cellular biology; Cholangiocarcinoma; Chronic; Clinic; Communication Research; Data; Development Plans; Digestive System Disorders; Disease; Environment; Goals; Grant; Hepatobiliary; Human; Inflammation; Inflammatory; Lead; MCL1 protein; Malignant Epithelial Cell; Malignant Neoplasms; Mentors; MicroRNAs; Mitochondria; Molecular Biology; Molecular Models; Mutation; Pathway interactions; Proteins; Public Health; Regulation; Research; Research Personnel; Resistance; Role; Techniques; Testing; Training; Transcript; Translational Research; age related; bile duct; cancer cell; career; career development; chemotherapy; cholangiocyte; cytokine; experience; improved; molecular modeling; new therapeutic target; novel; overexpression; protein expression; skills; stem; tumor

DESCRIPTION (provided by applicant): My long-term career goals are to become an independent investigator studying the role of cell death in diseases of the Gl tract. My previous research experience in cell and molecular biology related to cell death included my doctoral dissertation on the role of mitochondrial mutations in age-related disease. I have identified a highly successful mentor in the area of cell death in hepatobiliary diseases, including cancer. The lab is in the conducive environment of the Center for Basic Research in Digestive Diseases, at Mayo Clinic, Rochester. This proposal outlines a testable hypothesis to define the role of microRNAs as regulators of apoptosis in cholangiocarcinoma, a malignant neoplasm that occurs in the presence of chronic inflammation of bile ducts. The current proposal will allow the candidate to change research directions by providing advanced training in hepatobiliary disease and cancer biology as it relates to cell death. Short-term career goals include advanced training in cancer biology and Gl-related translational research. These immediate goals will be achieved by the career development plan, including research, communication skills, and mentoring training. The research plan stems from our previous observations that the antiapoptotic protein Mcl-1 is over-expressed in human cholangiocarcinoma and is a key regulator of cell death. One potential mechanism of Mcl-1 regulation is through microRNAs, short RNAs that act as sequence-specific silencers of protein expression. Our preliminary data shows that Mcl-1 expression is a predicted target of mir-29b, and antagonism of mir-29 increases Mcl-1 protein expression. Further, Mcl-1 is over-expressed in cholangio- carcinoma cells while mir-29b expression is decreased. Finally, blocking mir-29 protects cholangiocytes from cell death. These extensive and original observations support the OVERALL HYPOTHESIS of this grant that dysregulated expression of mir-29b results in over expression of Mcl-1, rendering this cancer apoptosis resistant. To address this hypothesis, we have established advanced cell culture models and molecular techniques to detect and alter expression of microRNAs. This project is relevant to public health as it addresses how bile duct cancers resist cell death, and as such resist chemotherapy. In addition, by testing a new pathway governing cancer cell death, we may define a novel set of targets to improve treatment of this devastating disease.

描述（由申请人提供）： 我的长期职业目标是成为一名独立研究者，研究细胞死亡在胃肠道疾病中的作用。我之前在与细胞死亡相关的细胞和分子生物学方面的研究经验包括我的博士论文，主题是线粒体突变在与年龄相关的疾病中的作用。我在肝胆疾病（包括癌症）的细胞死亡领域找到了一位非常成功的导师。该实验室位于罗切斯特梅奥诊所消化疾病基础研究中心的有利环境中。该提案概述了一个可检验的假设，以定义 microRNA 作为胆管癌细胞凋亡调节剂的作用，胆管癌是一种在胆管慢性炎症存在时发生的恶性肿瘤。目前的提案将允许候选人通过提供肝胆疾病和与细胞死亡相关的癌症生物学方面的高级培训来改变研究方向。短期职业目标包括癌症生物学和胃肠道相关转化研究方面的高级培训。这些近期目标将通过职业发展计划来实现，包括研究、沟通技巧和指导培训。该研究计划源于我们之前的观察，即抗凋亡蛋白Mcl-1在人胆管癌中过度表达，并且是细胞死亡的关键调节因子。 Mcl-1 调节的一种潜在机制是通过 microRNA，即充当蛋白质表达的序列特异性沉默子的短 RNA。我们的初步数据表明，Mcl-1 表达是 mir-29b 的预测目标，并且 mir-29 的拮抗作用增加了 Mcl-1 蛋白表达。此外，Mcl-1 在胆管癌细胞中过度表达，而 mir-29b 表达减少。最后，阻断 mir-29 可保护胆管细胞免于细胞死亡。这些广泛而原始的观察结果支持了本次资助的总体假设，即 mir-29b 的表达失调会导致 Mcl-1 的过度表达，从而使这种癌症具有细胞凋亡抵抗性。为了解决这一假设，我们建立了先进的细胞培养模型和分子技术来检测和改变 microRNA 的表达。该项目与公共卫生相关，因为它探讨了胆管癌如何抵抗细胞死亡，从而抵抗化疗。此外，通过测试控制癌细胞死亡的新途径，我们可以定义一组新的目标来改善这种破坏性疾病的治疗。