Androgen and MicroRNAs in Prostate Cancer

前列腺癌中的雄激素和 MicroRNA

• 批准号: 8038178
• 负责人: Anindya Dutta
• 金额: $ 31.96万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8038178
• 关键词: Advanced Malignant Neoplasm; Androgen Antagonists; Androgens; Bioinformatics; Biological Assay; Biological Markers; Cancer cell line; Castration; Cell Culture Techniques; Cell Line; Cells; Cessation of life; Chromatin Remodeling Factor; Clinical; Cloning; Diagnosis; Ensure; Epithelial Cells; Family; Fractionation; Future; Gene Expression; Gene Targeting; Genes; Genetic Transcription; In Vitro; Kinetics; Lead; Libraries; Malignant - descriptor; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Messenger RNA; Methods; MicroRNAs; Molecular Analysis; Mus; Neoplasm Metastasis; Nucleotides; Phenotype; Polyribosomes; Process; Prostate; RNA; Relative (related person); Repression; Research Institute; SMARCA5 gene; SMARCD1 gene; Sampling; Signal Transduction; Small RNA; Staging; Testing; Therapeutic; Therapeutic Studies; Tissues; Universities; Urologic Cancer; Virginia; Xenograft procedure; anticancer research; cancer cell; cell type; cellular engineering; gene induction; gene repression; improved; in vivo; interest; locked nucleic acid; mTOR protein; male; malignant phenotype; migration; new technology; novel; programs; research study; response; tool; tumor; tumor progression

DESCRIPTION (provided by applicant): MicroRNAs are 17-24 nucleotide long single-stranded RNA molecules that profoundly impact the gene expression program of a cell by annealing to and suppressing the expression of many target genes. There are hundreds of microRNAs and different cell types express different levels of specific microRNAs. It has recently become apparent that changes in expression of microRNAs are involved in different types of malignancies. In addition, new families of short RNAs, like pi-RNAs, rasi-RNAs and tRFs are being discovered that are not related to microRNAs but also impact the phenotype of a cell. As part of the Mellon Urological Cancer Research Institute at the University of Virginia we are interested in studying and reversing prostate cancer progression. Prostate cancers are highly dependent on androgens for their proliferation. Androgens act on prostate epithelial cells by changing the gene expression program, but their effects on microRNA and other short RNA expression have not been studied. In addition, although prostate cancers are treated with and initially very responsive to anti-androgens, they usually recur as androgen-independent cancers that eventually metastasize and lead to death. To reverse this progression, we have to understand the mechanism by which prostate cancer cells become androgen-independent and acquire other features of progression. In this proposal we will clone and sequence by Solexa/Illumina libraries of small RNAs from prostate cancer cells to identify microRNAs and other short RNAs that are (a) regulated by androgens and (b) involved in prostate cancer progression. We will also follow the changes in known microRNAs by hybridization to locked-nucleic acid microarrays. These results will not only identify new short RNAs involved in prostate cancer progression, but also test the hypothesis that androgen-repressed short RNAs are often also repressed during progression of prostate cancer. We will then test the hypothesis that microRNAs repressed during prostate cancer progression contribute to the phenotype of progression, both in vitro and in vivo, starting with four microRNAs (in two families) that have already been validated in preliminary results. The final test of the hypothesis will be the identification of targets of these microRNAs whose de-repression during cancer progression leads to worsening of the malignant phenotype. We will identify the targets of two key microRNA families involved in cancer progression by a novel combination of combination of experiments and computational target prediction, starting with eight targets that have already passed the experimental filters. Thus new technologies, short RNA cloning and ultra-high throughput sequencing, and new combinations of assays for identification of relevant targets will be applied to the molecular analysis of prostate cancer progression and the function of key microRNAs in this process dissected. The results are expected to open new avenues of research, therapy and diagnosis of prostate cancers. PUBLIC HEALTH RELEVANCE: MicroRNAs and short RNAs that are repressed by androgens will open a new chapter in how androgens regulate gene expression. Specific microRNAs and short RNAs that are also repressed during prostate cancer progression can be increased for therapeutic purposes. Studying the effects of these microRNAs (and short RNAs) on target genes will reveal how progression of prostate cancer is accompanied by repression of microRNAs to de-repress specific target genes, rendering the cancer cells less dependent on androgens, more proliferative, migratory and invasive and more metastatic. Thus, these androgen-repressed and progression-repressed microRNAs and non-micro-short RNAs will be useful both as biomarkers of cancer progression and as tools to identify genes whose de-repression is critical for cancer progression. Two methods will be developed here that will improve the study of microRNAs in cancer: (1) The combination of cloning/sequencing with locked-nucleic acid microarrays to obtain a well validated list of microRNAs that are changed during cancer progression. (2) The combination of bioinformatics, microarrays of mRNAs and polyribosome fractionation of mRNAs to produce a list of targets with a high true-positive rate of being direct targets of the microRNAs.

描述（由申请人提供）：MicroRNA是17-24个核苷酸长的单链RNA分子，其通过与许多靶基因退火并抑制其表达而深刻地影响细胞的基因表达程序。有数百种microRNA，不同的细胞类型表达不同水平的特定microRNA。最近已经变得明显的是，microRNA表达的变化涉及不同类型的恶性肿瘤。此外，还发现了新的短RNA家族，如pi-RNA、rasi-RNA和tRF，它们与微小RNA无关，但也影响细胞的表型。作为弗吉尼亚大学梅隆泌尿系癌症研究所的一部分，我们对研究和逆转前列腺癌进展感兴趣。前列腺癌的增殖高度依赖雄激素。雄激素通过改变基因表达程序作用于前列腺上皮细胞，但它们对microRNA和其他短RNA表达的影响尚未研究。此外，尽管前列腺癌用抗雄激素治疗并且最初对抗雄激素非常敏感，但它们通常作为雄激素非依赖性癌症复发，最终转移并导致死亡。为了逆转这种进展，我们必须了解前列腺癌细胞变得不依赖雄激素并获得其他进展特征的机制。在本提案中，我们将通过Solexa/Illumina对来自前列腺癌细胞的小RNA文库进行克隆和测序，以鉴定（a）受雄激素调节和（B）参与前列腺癌进展的microRNA和其他短RNA。我们还将通过与锁定核酸微阵列杂交来跟踪已知microRNA的变化。这些结果不仅将鉴定参与前列腺癌进展的新的短RNA，而且还将检验雄激素抑制的短RNA在前列腺癌进展期间通常也被抑制的假设。然后，我们将测试这一假设，即在前列腺癌进展过程中被抑制的microRNA有助于体外和体内进展的表型，从四种microRNA（两个家族）开始，这些microRNA已经在初步结果中得到验证。该假设的最终测试将是鉴定这些microRNA的靶点，这些microRNA在癌症进展期间的去抑制导致恶性表型的恶化。我们将通过实验和计算目标预测的新组合来确定参与癌症进展的两个关键microRNA家族的目标，从已经通过实验过滤器的八个目标开始。因此，新技术，短RNA克隆和超高通量测序，以及用于鉴定相关靶点的新的测定组合将被应用于前列腺癌进展的分子分析，并解剖了该过程中关键microRNA的功能。这些结果有望为前列腺癌的研究、治疗和诊断开辟新的途径。 公共卫生相关性：被雄激素抑制的microRNA和短RNA将为雄激素如何调节基因表达开辟新的篇章。在前列腺癌进展过程中也被抑制的特定microRNA和短RNA可以出于治疗目的而增加。研究这些microRNA（和短RNA）对靶基因的影响将揭示前列腺癌的进展如何伴随着microRNA的抑制以解除特定靶基因的抑制，使癌细胞对雄激素的依赖性降低，更具增殖性，迁移性和侵袭性，更具转移性。因此，这些雄激素抑制和进展抑制的微小RNA和非微小短RNA将可用作癌症进展的生物标志物和鉴定其去抑制对癌症进展至关重要的基因的工具。本文将开发两种方法来改善癌症中microRNA的研究：（1）将克隆/测序与锁定核酸微阵列相结合，以获得在癌症进展期间发生变化的microRNA的有效列表。(2)结合生物信息学、mRNA微阵列和mRNA的多核糖体分级分离，产生一系列具有高真阳性率的靶标，这些靶标是microRNA的直接靶标。