Functional characterization of the long non-coding RNA Scirocco in development an

长链非编码 RNA Scirocco 开发中的功能表征

• 批准号: 8434861
• 负责人: FRANK U SAUER
• 金额: $ 29.04万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8434861
• 关键词: A Mouse; Amino Acid Motifs; Attenuated; Biological Assay; Breast Cancer Cell; Cancer Etiology; Cancerous; Cell Differentiation process; Cell Proliferation; Cells; ChIP-seq; Chromatin; Chromatin Structure; Data; Development; Disease; EZH2 gene; Ectopic Expression; Epigenetic Process; Functional RNA; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Homeobox Genes; Human; In Vitro; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Mammary Neoplasms; Mammary gland; Mediating; Mus; Proteins; RNA; RNA-Binding Proteins; Recruitment Activity; Research; Role; SCID Mice; Signal Transduction Pathway; Transcript; Tretinoin; Tumor-Derived; Xenograft procedure; base; cancer stem cell; design; human disease; in vivo; insight; malignant breast neoplasm; migration; mutant; novel; therapeutic development; tumor progression

DESCRIPTION (provided by applicant): Long, non-coding RNAs (lncRNAs) have emerged as important regulators of chromatin structure and function. Vertebrate lncRNAs have been closely associated with epigenetic silencing by recruiting epigenetic repressors to target genes. The specific hypothesis behind the proposed research is that the novel lncRNA Scirocco (SCIR) controls cell differentiation and proliferation in development and cancer. The specific aims are designed to provide a comprehensive assessment of the role of SCIR in cell differentiation and proliferation in development and disease. Aim 1. To dissect the interaction of SCIR with EZH2: SCIR interacts with the EZH2 subunit of PRC2 in vitro and the interaction of SCIR with EZH2 evicts PRC2 from chromatin. How SCIR interacts with EZH2 remains unclear. The RNA and protein motifs involved in the association of SCIR with EZH2 will be uncovered by in vitro RNA-protein binding assays, which investigate the interaction of EZH2 with mutant SCIR transcripts and vice versa. To assess whether the identified RNA and protein motifs support the EZH2-SCIR interaction in vivo, we shall investigate the association of EZH2 with SCIR in cells, which express mutant SCIR transcripts or mutant EZH2 proteins. Aim 2. To dissect the role of SCIR in development: Our data suggest that all-trans retinoic acid mediated activation of SCIR results in activation of HOXA1 and HOXA2 and culminates in cell differentiation and proliferation. How RA activates SCIR transcription and how the SCIR-mediated activation of HOXA1 and HOXA2 controls cell proliferation and differentiation remains unknown. We shall combine RNA- and ChIP- sequencing assays with functional assays to identify target genes for HOXA1 and HOXA2. To uncover the role of SCIR in development, we shall investigate the development SCIR-deficient cells and mice. We shall investigate whether the inactivation of components of the RA signal transduction pathway attenuates SCIR transcription. Aim 3: To dissect the role of SCIR in cancer: We have detected aberrant transcription of SCIR in primary breast and lung cancer and demonstrated that SCIR controls the proliferation, migration and invasiveness of lung and breast cancer cells. Our results suggest that anomalous SCIR transcription is involved in initiation and progression of cancer. We shall investigate whether the ectopic expression of SCIR causes cancer in mice. Accomplishing the three aims of this proposal will support he role of SCIR in development and cancer by providing evidence that the lncRNA SCIR directly interacts with epigenetic repressor EZH2 (Aim 1), elucidating the role of SCIR in cell differentiation and proliferation (Aim 2), and uncovering the role of SCIR in initiation and progression of cancer (Aim 3).

描述(申请人提供)：长的非编码RNA(LncRNAs)已经成为染色质结构和功能的重要调节因子。脊椎动物的lncRNAs通过招募表观遗传抑制因子来靶向基因，从而与表观遗传沉默密切相关。这项研究背后的具体假设是，新的lncRNA Scirocco(SCIR)在发育和癌症过程中控制细胞的分化和增殖。这些具体目标旨在全面评估SCIR在发育和疾病过程中细胞分化和增殖方面的作用。目的1.分析SCIR与EZH2的相互作用：SCIR在体外与PRC2的EZH2亚基相互作用，并将PRC2从染色质中剔除。SCIR如何与EZH2互动仍不清楚。参与SCIR与EZH2结合的RNA和蛋白质基序将通过体外RNA-蛋白质结合分析来揭示，该分析研究EZH2与突变的SCIR转录本之间的相互作用，反之亦然。为了评估所识别的RNA和蛋白质基序是否支持体内EZH2-SCIR的相互作用，我们将研究EZH2与细胞中表达突变的SCIR转录本或突变的EZH2蛋白的相关性。目的2.分析SCIR在发育中的作用：我们的数据表明，全反式维甲酸介导的SCIR激活导致HOXA1和HOXA2的激活，最终导致细胞分化和增殖。RA如何激活SCIR转录，以及SCIR介导的HOXA1和HOXA2的激活如何控制细胞的增殖和分化仍是未知的。我们将结合RNA测序和芯片测序以及功能分析来确定HOXA1和HOXA2的靶基因。为了揭示SCIR在发育中的作用，我们将研究SCIR缺陷细胞和小鼠的发育。我们将研究RA信号转导通路组件的失活是否减弱了SCIR的转录。目的3：分析SCIR在癌症中的作用：我们在乳腺癌和肺癌中检测到了SCIR的异常转录，并证实了SCIR控制着肺癌和乳腺癌细胞的增殖、迁移和侵袭。我们的结果提示，SCIR转录异常参与了癌症的发生和发展。我们将调查SCIR的异位表达是否会导致小鼠癌症。实现本提案的三个目标将支持SCIR在发育和癌症中的作用，因为它提供了lncRNA SCIR与表观遗传抑制因子EZH2直接相互作用的证据(目标1)，阐明了SCIR在细胞分化和增殖中的作用(目标2)，并揭示了SCIR在癌症的发生和发展中的作用(目标3)。