Deregulation of long noncoding RNAs in cancer

癌症中长非编码RNA的失调

• 批准号: 10674961
• 负责人: Nadya M Dimitrova
• 金额: $ 49.57万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674961
• 关键词: Acceleration; Acute; Address; Affect; Animal Model; Animals; Binding Proteins; Bioinformatics; Biology; CRISPR-mediated transcriptional activation; CRISPR/Cas technology; Cell Line; Cell model; Cells; Chemicals; Chromatin; Collaborations; Development; Disease; Disease Progression; Dissociation; Dominant-Negative Mutation; Down-Regulation; Doxycycline; Effectiveness; Event; Gene Expression; Genes; Genetic Models; Genetic Transcription; Goals; Human; Inflammatory; KRAS2 gene; Knock-out; Lung Adenocarcinoma; MALAT1 gene; Malignant Neoplasms; Mediating; Mediator; Modeling; Molecular; Molecular Biology; Mus; Neoplasm Metastasis; Patients; Predisposition; Prognosis; RNA Splicing; Regulation; Role; Series; Signal Transduction; System; TP53 gene; Technology; Testing; Therapeutic; Time; Tumor Promotion; Untranslated RNA; Up-Regulation; Work; cohort; conditional knockout; cytokine; design; differential expression; experimental study; feasibility testing; gain of function; innovation; mouse model; novel; novel therapeutic intervention; overexpression; patient prognosis; pleiotropism; posttranscriptional; programs; therapeutic target; tumor; tumor initiation; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic

Project Summary/Abstract Aberrantly high expression of the long noncoding RNA (lncRNA) Malat1 is strongly associated with lung adenocarcinoma (LUAD) progression and poor patient prognosis and has been pursued as a potential therapeutic target. However, these correlative observations do not reveal whether the aberrant expression of Malat1 is a driver of tumorigenesis and what are the mechanisms by which Malat1 overexpression promotes disease progression. In preliminary studies, we developed an innovative CRISPR activation (CRISPRa) system that successfully models Malat1 overexpression in patient-derived cell lines and autochthonous murine models of LUAD. Strikingly, we uncovered that tumor-specific overexpression of Malat1 at the time of tumor initiation is sufficient to accelerate the progression of murine LUAD to aggressive and metastatic disease, demonstrating that Malat1 overexpression is a driver of cancer development. We further determined that Malat1 overexpression leads to pleiotropic effects on the tumor microenvironment through the altered expression of cytokines and stromal factors. Based on these promising findings, our central hypothesis is that progressive accumulation of Malat1 in LUAD drives the development of aggressive disease through the activation of a metastasis-promoting gene expression program and the establishment of a pro-metastatic tumor microenvironment. In Aim 1, we describe our use of advanced, temporally controlled mouse models to clarify the stage of tumorigenesis when overexpression of Malat1 promotes tumor progression, the requirement for sustained Malat1 overexpression, and the window of opportunity when downregulation of Malat1 may have a therapeutic impact. Aim 2 seeks to investigate the non-cell autonomous effects of Malat1 overexpression in the establishment of a pro-metastatic niche. We propose to perform detailed characterization of Malat1-dependent changes in the tumor stroma at different stages of tumorigenesis as well as use genetic models to explore the contributions of candidate secreted factors, such as inflammatory cytokines. Finally, Aim 3 outlines our plans to elucidate the molecular mechanism by which overexpressed Malat1 alters the expression of tumor microenvironment effectors. We propose to utilize state-of-the-art molecular biology approaches and work with long-term collaborators, who are experts in bioinformatics and chemical biology, to determine whether Malat1 overexpression affects target genes at the transcriptional or post-transcriptional level, through dominant negative or gain-of-function mechanisms, and through direct or indirect activities. In summary, this proposal outlines a highly innovative experimental and conceptual framework for dissecting the poorly understood role of overexpressed Malat1 as a driver in LUAD. Beyond Malat1, the broad significance of this work lies in its potential to elucidate how aberrantly expressed lncRNAs modulate cancer development and to uncover whether targeting aberrantly expressed lncRNAs might hold a therapeutic promise.

项目总结/摘要 长链非编码RNA（lncRNA）Malat 1的异常高表达与肺 腺癌（LUAD）进展和患者预后差，并已被视为潜在的治疗方法。 治疗靶点然而，这些相关的观察结果并没有揭示是否异常表达， Malat 1是肿瘤发生的驱动因素，Malat 1过表达促进肿瘤发生的机制是什么？ 疾病进展。在初步研究中，我们开发了一种创新的CRISPR激活（CRISPRa） 在患者来源的细胞系和本地小鼠中成功模拟Malat 1过表达的系统 LUAD模型。令人惊讶的是，我们发现肿瘤特异性Malat 1的过度表达在肿瘤发生时， 起始足以加速鼠LUAD向侵袭性和转移性疾病的进展， 这表明Malat 1过表达是癌症发展的驱动因素。我们进一步确定， Malat 1过表达通过改变肿瘤微环境导致肿瘤微环境的多效性效应。 细胞因子和基质因子的表达。基于这些有希望的发现，我们的中心假设是， Malat 1在LUAD中的进行性积累通过以下途径驱动侵袭性疾病的发展： 转移促进基因表达程序的激活和促转移肿瘤的建立 微环境在目标1中，我们描述了我们使用先进的，时间控制的小鼠模型来阐明 在肿瘤发生的阶段，Malat 1的过表达促进肿瘤进展， 持续的Malat 1过表达，以及Malat 1下调的机会窗口可能具有 治疗效果目的2旨在研究Malat 1过表达在细胞中的非细胞自主作用。 建立促转移的小生境。我们建议对Malat 1依赖的 在肿瘤发生的不同阶段肿瘤间质的变化，以及使用遗传模型来探索 候选分泌因子的贡献，如炎性细胞因子。最后，目标3概述了我们的计划， 阐明过表达的Malat 1改变肿瘤表达的分子机制 微环境效应器我们建议利用最先进的分子生物学方法， 长期合作者，谁是生物信息学和化学生物学专家，以确定是否Malat 1 过表达在转录或转录后水平上影响靶基因，通过显性表达， 消极或功能获得机制，并通过直接或间接的活动。总而言之，这一提议 概述了一个高度创新的实验和概念框架，用于解剖人们知之甚少的角色， 过表达的Malat 1作为LUAD的驱动因子。除了马拉特1，这项工作的广泛意义在于其 有可能阐明异常表达的lncRNA如何调节癌症的发展， 靶向异常表达的lncRNA是否有治疗前景。