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: 8236460
• 负责人: FRANK U SAUER
• 金额: $ 30.96万
• 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/8236460
• 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). PUBLIC HEALTH RELEVANCE: Epigenetic mechanisms establish and maintain the identity of cells during development. Errors in epigenetic mechanisms have been correlated with various human diseases such as cancer. Long, non-coding RNAs (ncRNAs) have emerged as important regulators of epigenetic phenomena. The proposed project will assess the role and function of the ncRNA Scirocco in epigenetic gene expression in development and cancer. Accomplishing the described project will provide novel insights into epigenetic mechanisms and open novel avenues for the development of therapeutic assays that attenuate cancer.

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因子。脊椎动物 lncRNA 通过向靶基因招募表观遗传抑制子而与表观遗传沉默密切相关。这项研究背后的具体假设是，新型 lncRNA Scirocco (SCIR) 控制着发育和癌症中的细胞分化和增殖。具体目标旨在全面评估 SCIR 在发育和疾病中细胞分化和增殖中的作用。目标 1. 剖析 SCIR 与 EZH2 的相互作用：SCIR 在体外与 PRC2 的 EZH2 亚基相互作用，并且 SCIR 与 EZH2 的相互作用将 PRC2 从染色质中驱逐。 SCIR 如何与 EZH2 相互作用仍不清楚。参与 SCIR 与 EZH2 关联的 RNA 和蛋白质基序将通过体外 RNA-蛋白质结合测定来揭示，该测定研究 EZH2 与突变 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SCIR在表达突变型SCIR转录本或突变型EZH2蛋白的细胞中的关联。目标 2. 剖析 SCIR 在发育中的作用：我们的数据表明，全反式视黄酸介导的 SCIR 激活导致 HOXA1 和 HOXA2 激活，并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 激活如何控制细胞增殖和分化仍然未知。我们将 RNA 和 ChIP 测序分析与功能分析相结合，以确定 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 信号转导通路成分的失活是否会减弱 SCIR 转录。目标 3：剖析 SCIR 在癌症中的作用：我们在原发性乳腺癌和肺癌中检测到 SCIR 的异常转录，并证明 SCIR 控制肺癌和乳腺癌细胞的增殖、迁移和侵袭。我们的结果表明异常 SCIR 转录参与癌症的发生和进展。我们将研究 SCIR 的异位表达是否会导致小鼠癌症。实现该提案的三个目标将通过提供lncRNA SCIR直接与表观遗传阻遏物EZH2相互作用的证据来支持SCIR在发育和癌症中的作用（目标1），阐明SCIR在细胞分化和增殖中的作用（目标2），并揭示 SCIR 在癌症发生和进展中的作用（目标 3）。 公共卫生相关性：表观遗传机制在发育过程中建立并维持细胞的身份。表观遗传机制的错误与癌症等多种人类疾病有关。长链非编码 RNA (ncRNA) 已成为表观遗传现象的重要调节因子。拟议的项目将评估 ncRNA Scirocco 在发育和癌症表观遗传基因表达中的作用和功能。完成所描述的项目将为表观遗传机制提供新的见解，并为开发减轻癌症的治疗方法开辟新途径。