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(申请人提供)：microRNAs是17-24个核苷酸长的单链RNA分子，通过使许多目标基因的表达受到抑制而对细胞的基因表达程序产生深远的影响。有数百种microRNAs，不同的细胞类型表达不同水平的特定microRNAs。近来发现，microRNAs表达的变化与不同类型的恶性肿瘤有关。此外，新的短RNA家族，如pi-RNAs、Rasi-RNAs和TRFs正在被发现，它们与microRNAs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的进展很感兴趣。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，但其对microRNA和其他短RNA表达的影响尚未被研究。此外，尽管前列腺癌接受了抗雄激素的治疗，并且最初对其非常敏感，但它们通常会复发为雄激素非依赖性癌症，最终会转移并导致死亡。为了逆转这一进程，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。在这项建议中，我们将通过Solexa/Illumina克隆和测序前列腺癌细胞中的小RNA，以确定(A)受雄激素调控和(B)参与前列腺癌进展的microRNAs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microRNAs的变化。这些结果不仅将识别与前列腺癌进展有关的新的短RNA，而且还将检验雄激素抑制的短RNA通常在前列腺癌进展过程中也被抑制的假说。然后，我们将测试这一假设，即前列腺癌进展过程中被抑制的microRNA在体外和体内都有助于进展的表型，首先是已经在初步结果中得到验证的四个microRNA(在两个家庭中)。对这一假说的最终检验将是确定这些microRNAs的靶点，这些microRNAs在癌症进展过程中的抑制会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，从已经通过实验过滤器的八个目标开始，确定参与癌症进展的两个关键microRNA家族的目标。因此，新的技术、短RNA克隆和超高通量测序以及鉴定相关靶点的新的检测组合将被应用于前列腺癌进展的分子分析和关键microRNAs在这一过程中的功能剖析。这一结果有望为前列腺癌的研究、治疗和诊断开辟新的途径。 与公共健康相关：受雄激素抑制的微小RNA和短RNA将开启雄激素调控基因表达的新篇章。在前列腺癌进展过程中也被抑制的特定的microRNA和短RNA可以出于治疗目的而增加。研究这些微RNA(和短RNA)对靶基因的影响将揭示前列腺癌的进展是如何伴随着抑制特定靶基因的微RNA而来的，从而使癌细胞对雄激素的依赖性更低，更具增殖、迁移和侵袭性，更具转移性。因此，这些雄激素抑制和进展抑制的microRNA和非微型短RNA将是有用的，既可以作为癌症进展的生物标志物，也可以作为工具来识别其下调抑制对癌症进展至关重要的基因。这里将开发两种方法来改进癌症中的microRNAs的研究：(1)克隆/测序与锁定核酸微阵列相结合，以获得在癌症进展过程中发生变化的经过充分验证的microRNAs列表。(2)生物信息学、mRNAs微阵列和mRNAs多聚核糖体分离相结合，以产生具有高真阳性率的靶点列表，成为microRNAs的直接靶标。