MicroRNA Biogenesis and the Cancer Proteome

MicroRNA 生物发生和癌症蛋白质组

• 批准号: 7431456
• 负责人: MARK D JOHNSON
• 金额: $ 262.5万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7431456
• 关键词: Accounting; Acids; Actins; Address; Affect; Affinity; Agreement; Algorithms; Anabolism; Anecdotes; Animals; Apoptosis; Area; Attention; Award; Behavior; Bibliography; Binding; Biogenesis; Bioinformatics; Biological; Biological Assay; Biological Process; Biomedical Computing; Biostatistical Methods; Biotin; Brain; Brain Glioblastoma; Brain Neoplasms; Breast; Camptothecin; Cancer Biology; Cancer Model; Cancer Patient; Cancer Survivorship; Caring; Caspase; Categories; Cations; Cell Cycle; Cell Cycle Progression; Cell Death; Cell Differentiation process; Cell Line; Cell Proliferation; Cells; Cellular Assay; Cellular biology; Cessation of life; Characteristics; Chemicals; Chromatography; Chromosome Pairing; Cleaved cell; Clinic; Clinical; Clinical Data; Clinical Research; Cluster Analysis; Code; Collaborations; Color; Complex; Computer software; Computers; Condition; Control Animal; Core Facility; Coupled; Creativeness; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinases; Cyclotrons; DICER1 gene; DNA; DNA Damage; DNA Integration; DNA Sequence; DNA analysis; Data; Data Analyses; Data Set; Defect; Development; Disease regression; Disruption; Distress; Doctor of Philosophy; Down-Regulation; Dyes; Elements; Embryo; Exhibits; Failure; Family; Figs - dietary; Fostering; Fourier Transform; Freezing; Frequencies; Functional RNA; Gene Cluster; Gene Expression; Gene Mutation; Gene Order; Genes; Genetic; Genets; Genome; Genome Mappings; Genomics; Glioblastoma; Glioma; Glutamates; Growth; Guidelines; HDAC1 gene; Hematoxylin and Eosin Staining Method; Heterogeneity; Histology; Hospital Records; Hospitals; Human; Human Resources; Immigration; Implant; In Vitro; Individual; Institution; Ions; Isotopes; Isotopically-Coded Affinity Tagging; Journals; Kolmogorov-Smirnov Test; Label; Laboratories; Lead; Light; Liquid Chromatography; Location; Lung; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of lung; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Mediating; Mentors; Messenger RNA; Metabolic; Methodology; Methods; Methylation; MicroRNAs; Microarray Analysis; Mitogens; Modeling; Molecular; Molecular Profiling; Molecular and Cellular Biology; Mus; Mutation; National Cancer Institute; Nature; Nervous system structure; Neurobiology; Neurons; Neurosurgeon; Normal Cell; Numbers; Oncogenes; Operative Surgical Procedures; Outcome; Output; Ovarian; PDGFRA gene; Paper; Pathway Analysis; Pathway interactions; Patients; Pattern; Peptides; Physical Map of the Human Genome; Play; Polymerase Chain Reaction; Positioning Attribute; Primary Neoplasm; Probability; Process; Production; Protein Overexpression; Proteins; Proteome; Proteomics; Protocols documentation; Publishing; RLK5-associated protein phosphatase; RNA; RNA Polymerase III; RNA Splicing; RNA analysis; Range; Rate; Rattus; Recording of previous events; Regression Analysis; Regulation; Relative (related person); Reporting; Research; Research Ethics Committees; Research Personnel; Residencies; Resolution; Resources; Role; Sampling; Scanning; Science; Scientist; Seminal; Signal Pathway; Signal Transduction; Skin; Slide; Solid; Source; Specificity; Specimen; Spottings; Staging; Students; Subfamily lentivirinae; Subgroup; Survival Analysis; Synapses; System; TP53 gene; Techniques; Technology; Testing; Time; Tissue Banks; Tissue Extracts; Tissue Sample; Tissues; Training; Translational Research; Translations; Trypsin; Tumor Suppressor Proteins; Tumor Tissue; Tumorigenicity; Universities; Up-Regulation; Validation; Washington; Week; Western Blotting; Width; Woman; Work; abstracting; animal care; base; cancer cell; cancer genome; carcinogenesis; career; cell growth; cell motility; cellular transduction; clinically relevant; comparative; cyanine dye 5; cyclin-dependent kinase 6; density; fight against; follow-up; forging; frontier; functional genomics; genome-wide analysis; hazard; human DICER1 protein; human subject; improved; in vitro Assay; in vitro Model; in vivo; innovation; insight; interest; mRNA Expression; mRNA Stability; mass spectrometer; medical schools; member; metaplastic cell transformation; mouse model; neoplastic cell; neuron apoptosis; neuron loss; neurosurgery; novel; novel therapeutics; outcome forecast; peripheral blood; physical mapping; postnatal; programs; protein expression; research study; response; reversed phase chromatography; size; small hairpin RNA; statistics; success; synthetic protein; therapeutic target; therapy outcome; tool; tumor; tumorigenesis; vector; vector control; willingness; Accounting; Acids; Actins; Address; Affect; Affinity; Agreement; Algorithms; Anabolism; Anecdotes; Animals; Apoptosis; Area; Attention; Award; Behavior; Bibliography; Binding; Biogenesis; Bioinformatics; Biological; Biological Assay; Biological Process; Biomedical Computing; Biostatistical Methods; Biotin; Brain; Brain Glioblastoma; Brain Neoplasms; Breast; Camptothecin; Cancer Biology; Cancer Model; Cancer Patient; Cancer Survivorship; Caring; Caspase; Categories; Cations; Cell Cycle; Cell Cycle Progression; Cell Death; Cell Differentiation process; Cell Line; Cell Proliferation; Cells; Cellular Assay; Cellular biology; Cessation of life; Characteristics; Chemicals; Chromatography; Chromosome Pairing; Cleaved cell; Clinic; Clinical; Clinical Data; Clinical Research; Cluster Analysis; Code; Collaborations; Color; Complex; Computer software; Computers; Condition; Control Animal; Core Facility; Coupled; Creativeness; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinases; Cyclotrons; DICER1 gene; DNA; DNA Damage; DNA Integration; DNA Sequence; DNA analysis; Data; Data Analyses; Data Set; Defect; Development; Disease regression; Disruption; Distress; Doctor of Philosophy; Down-Regulation; Dyes; Elements; Embryo; Exhibits; Failure; Family; Figs - dietary; Fostering; Fourier Transform; Freezing; Frequencies; Functional RNA; Gene Cluster; Gene Expression; Gene Mutation; Gene Order; Genes; Genetic; Genets; Genome; Genome Mappings; Genomics; Glioblastoma; Glioma; Glutamates; Growth; Guidelines; HDAC1 gene; Hematoxylin and Eosin Staining Method; Heterogeneity; Histology; Hospital Records; Hospitals; Human; Human Resources; Immigration; Implant; In Vitro; Individual; Institution; Ions; Isotopes; Isotopically-Coded Affinity Tagging; Journals; Kolmogorov-Smirnov Test; Label; Laboratories; Lead; Light; Liquid Chromatography; Location; Lung; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of lung; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Mediating; Mentors; Messenger RNA; Metabolic; Methodology; Methods; Methylation; MicroRNAs; Microarray Analysis; Mitogens; Modeling; Molecular; Molecular Profiling; Molecular and Cellular Biology; Mus; Mutation; National Cancer Institute; Nature; Nervous system structure; Neurobiology; Neurons; Neurosurgeon; Normal Cell; Numbers; Oncogenes; Operative Surgical Procedures; Outcome; Output; Ovarian; PDGFRA gene; Paper; Pathway Analysis; Pathway interactions; Patients; Pattern; Peptides; Physical Map of the Human Genome; Play; Polymerase Chain Reaction; Positioning Attribute; Primary Neoplasm; Probability; Process; Production; Protein Overexpression; Proteins; Proteome; Proteomics; Protocols documentation; Publishing; RLK5-associated protein phosphatase; RNA; RNA Polymerase III; RNA Splicing; RNA analysis; Range; Rate; Rattus; Recording of previous events; Regression Analysis; Regulation; Relative (related person); Reporting; Research; Research Ethics Committees; Research Personnel; Residencies; Resolution; Resources; Role; Sampling; Scanning; Science; Scientist; Seminal; Signal Pathway; Signal Transduction; Skin; Slide; Solid; Source; Specificity; Specimen; Spottings; Staging; Students; Subfamily lentivirinae; Subgroup; Survival Analysis; Synapses; System; TP53 gene; Techniques; Technology; Testing; Time; Tissue Banks; Tissue Extracts; Tissue Sample; Tissues; Training; Translational Research; Translations; Trypsin; Tumor Suppressor Proteins; Tumor Tissue; Tumorigenicity; Universities; Up-Regulation; Validation; Washington; Week; Western Blotting; Width; Woman; Work; abstracting; animal care; base; cancer cell; cancer genome; carcinogenesis; career; cell growth; cell motility; cellular transduction; clinically relevant; comparative; cyanine dye 5; cyclin-dependent kinase 6; density; fight against; follow-up; forging; frontier; functional genomics; genome-wide analysis; hazard; human DICER1 protein; human subject; improved; in vitro Assay; in vitro Model; in vivo; innovation; insight; interest; mRNA Expression; mRNA Stability; mass spectrometer; medical schools; member; metaplastic cell transformation; mouse model; neoplastic cell; neuron apoptosis; neuron loss; neurosurgery; novel; novel therapeutics; outcome forecast; peripheral blood; physical mapping; postnatal; programs; protein expression; research study; response; reversed phase chromatography; size; small hairpin RNA; statistics; success; synthetic protein; therapeutic target; therapy outcome; tool; tumor; tumorigenesis; vector; vector control; willingness

Recent studies in human cancers have revealed defects in microRNA biogenesis that promote tumor aggressiveness and decrease survival via mechanisms that are poorly understood. Much of the control of protein expression by microRNAs occurs without alterations in mRNA expression, and single microRNAs may target dozens of mRNAs in a tissue specific manner. Thus, this broad-based decrease in microRNA synthesis presents a challenge to those trying to understand its downstream molecular effects on carcinogenesis. Here we propose a new strategy that will comprehensively identify the mechanisms by which defects in microRNA biogenesis decrease cancer survivorship. This approach involves the integration of genome-scale high throughput quantitative mass spectrometry analysis of the cancer proteome with genome-wide microRNA, mRNA and DNA analyses of human brain tumors (glioblastomas) that have intact or defective microRNA biogenesis. Novel algorithms that incorporate clinical variables will be used to generate an integrated genome- scale view of identified differences, and clinically-related targets will be investigated further using in vivo and in vitro models. Preliminary studies in our laboratory using aCGH and mRNA microarrays revealed deletion at the DICER1 locus and decreased DICER1 mRNA expression in a subset of glioblastomas, and this correlated with decreased microRNA expression and survival. Genome- wide analysis identified numerous microRNAs that were differentially expressed between glioblastomas with low versus high DICER1 expression. Cdk6, which promotes cell cycle progression, was a predicted target of several of these microRNAs. DICER1 knockdown in glioblastoma cells increased cell growth and upregulated Cdk6 protein expression. Importantly, Cdk6 protein was overexpressed in glioblastomas with low DICER1 expression, suggesting that Cdk6 upregulation may be one mechanism by which defective microRNA biogenesis contributes to increased tumor aggressiveness. This innovative and ambitious project will integrate mass spectrometry proteomics, genomics and clinical variables to comprehensively identify the mechanisms underlying the decreased cancer survivorship associated with dysregulated microRNA biogenesis.

在人类癌症中的最新研究表明，microRNA生物发生缺陷，促进 肿瘤的侵略性和通过知之甚少的机制降低生存率。大部分 microRNA对蛋白表达的控制发生不会改变mRNA表达，并且 单个microRNA可以以组织特异性方式靶向数十个mRNA。因此，这个基于广泛的 MicroRNA合成的减少给那些试图了解其下游的人带来了挑战 分子对癌变的影响。在这里，我们提出了一种新策略，将全面 确定microRNA生物发生缺陷降低癌症生存的机制。 这种方法涉及基因组规模高吞吐量定量质量的整合 用全基因组microRNA，mRNA和DNA对癌症蛋白质组的光谱法分析 对具有完整或有缺陷的MicroRNA生物发生的人脑肿瘤（胶质母细胞瘤）的分析。 结合临床变量的新型算法将用于产生综合基因组 - 确定差异的比例视图，将进一步研究与临床相关的目标。 体内和体外模型。使用ACGH和mRNA微阵列在我们的实验室进行初步研究 在DICER1基因座处揭示了缺失，并在一个子集中降低了DICER1 mRNA表达 胶质母细胞瘤，这与microRNA表达和存活率降低相关。基因组 广泛的分析确定了许多在 胶质母细胞瘤低于高diCER1表达。 CDK6，促进细胞周期 进展是其中几种microRNA的预测靶标。 DICER1击倒 胶质母细胞瘤细胞增加细胞生长和上调CDK6蛋白表达。重要的是， CDK6蛋白在胶质母细胞瘤中过表达，DICER1表达低，表明 CDK6上调可能是一种机制，有缺陷的microRNA生物发生有助于 肿瘤侵略性提高。这个创新且雄心勃勃的项目将整合质量 光谱法蛋白质组学，基因组学和临床变量可全面识别 与失调的microRNA相关的癌症生存降低的机制 生物发生。