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（由申请人提供）：长，非编码RNA（LNCRNA）已成为染色质结构和功能的重要调节剂。脊椎动物LNCRNA与表观遗传沉默密切相关，通过将表观遗传阻遏物募集到靶基因。拟议的研究背后的特定假设是，新型的LNCRNA SCIROCCO（SCIR）控制了发育和癌症中的细胞分化和增殖。该特定目的旨在对SCIR在细胞分化和发育和疾病中的增殖中的作用进行全面评估。目的1。剖析SCIR与EZH2：SCIR的相互作用：Scir在体外与PRC2的EZH2亚基相互作用，而SCIR与EZH2与EZH2的相互作用从染色质中驱逐PRC2。 Scir如何与EZH2相互作用尚不清楚。体外RNA-蛋白结合测定法将发现与SCIR与EZH2缔合的RNA和蛋白基序，这些分析研究了EZH2与突变体Scir转录本的相互作用，反之亦然。为了评估所鉴定的RNA和蛋白质基序是否支持体内EZH2-SCIR相互作用，我们将研究EZH2与细胞中SCIR的关联，这些细胞表达突变体SCIR转录本或突变体EZH2蛋白。目的2。剖析SCIR在发展中的作用：我们的数据表明，全反式视黄酸介导的SCIR激活会导致Hoxa1和Hoxa2的激活以及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增殖和分化。我们将将RNA和芯片测序测定法与功能测定法相结合，以鉴定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缺陷型细胞和小鼠的发育。我们将研究RA信号转导途径的分量失活是否减弱SCIR转录。 AIM 3：剖析SCIR在癌症中的作用：我们已经检测到了原发性乳腺癌和肺癌中SCIR的异常转录，并证明SCIR控制了肺和乳腺癌细胞的增殖，迁移和侵入性。我们的结果表明，异常的SCIR转录参与癌症的开始和进展。我们将研究SCIR的异位表达是否会导致小鼠癌症。通过提供证据表明，LNCRNA SCIR直接与表观遗传抑制剂EZH2（AIM 1）相互作用，阐明Scir在细胞分化和增殖中的作用（AIM 2），并揭示Scir在癌症的起始和进展中的作用（AIM 3），实现了该提案的三个目标将支持SCIR在发育和癌症中的作用。