Non-coding RNA functions in tumor metastasis

非编码RNA在肿瘤转移中的作用

基本信息

批准号：
10531262
负责人：
Li Ma
金额：
$ 35.28万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10531262
关键词：
Affect Antisense RNA Back Binding Binding Proteins Biological Markers Breast Cancer Cell Breast Cancer Model Breast Cancer cell line Breast cancer metastasis CRISPR/Cas technology Cells Chromatin Clinical DNA Disseminated Malignant Neoplasm Family member Gene Deletion Gene Expression Profiling Genes Genetic Genetic Models Genetic Transcription Genetically Engineered Mouse Genomic DNA Goals Human Knock-out MALAT1 gene Malignant Neoplasms Mammary Neoplasms Mass Spectrum Analysis Mediating Metastasis Induction Metastatic Neoplasm to the Lung MicroRNAs Modeling Molecular Mus Neoplasm Metastasis Normal tissue morphology Phenotype RNA RNA purification Regulatory Element Reporting Research Role Small Interfering RNA Testing Transgenic Mice Transgenic Model Tumor Tissue Untranslated RNA Up-Regulation Validation Xenograft Model Xenograft procedure breast cancer progression experimental study gain of function genome editing genomic locus insertion/deletion mutation insight loss of function mouse model neoplastic cell novel overexpression promoter reverse genetics therapeutic target transcription factor transcriptome sequencing transgene expression transplant model tumor

项目摘要

Although great advances have been made in combatting cancer, particularly at its early stages, metastasis remains a formidable and frequently fatal challenge. It has become evident that non-coding RNAs, including microRNAs and long non-coding RNAs (lncRNAs), are components of molecular networks regulating metastasis. Some lncRNAs are known to have opposing functions to their genomic locus; for instance, opposite phenotypes have been reported from the lncRNA Haunt gene deletion and insertional inactivation, and interestingly, the Haunt gene deletion effect was due to the loss of the genomic DNA but not the loss of Haunt RNA. Thus, a major challenge in lncRNA research is whether phenotypes resulting from deleting or inactivating a lncRNA gene can be unequivocally attributed either to the loss of the lncRNA per se or to the loss of overlapping regulatory elements. MALAT1 (metastasis associated lung adenocarcinoma transcript 1) is among the most abundant and conserved lncRNAs in normal tissues, and has previously been described as a metastasis promoter. However, there is no evidence that the previously reported Malat1 gene deletion (which led to upregulation of multiple Malat1's adjacent genes) or antisense RNA (which has never been validated by rescue experiments or by MALAT1 knockout cells) effect was specific to Malat1 lncRNA loss. Unexpectedly, using a transcriptional terminator insertion strategy, we found that disrupting the Malat1 gene without altering the expression of its adjacent genes in a transgenic mouse model of breast cancer drastically promoted lung metastasis, and importantly, this phenotype was completely reversed by genetic add-back of Malat1. Moreover, CRISPR-Cas9- mediated knockout of MALAT1 in human breast cancer cells induced their metastatic ability, which was reversed by Malat1 re-expression. Conversely, overexpression of Malat1 suppressed breast cancer metastasis in both transgenic mice and xenograft models. Mechanistically, we used a recently developed chromatin isolation by RNA purification-mass spectrometry (ChIRP-MS) approach to identify TEAD family members as binding proteins for MALAT1 at the endogenous level from primary mammary tumor tissues, and discovered that MALAT1 binds, sequesters, and inactivates the pro-metastatic transcription factor TEAD. We also found an inverse correlation of MALAT1 levels with breast cancer progression and metastasis. Based on these important findings, we propose to comprehensively characterize the loss-of-function and gain-of-function effects of MALAT1 in breast cancer metastasis, using genetically engineered mouse models, transplantation models, syngeneic models, xenograft models, and CRISPR-Cas9 genome editing approaches (Specific Aim 1); we will also elucidate the mechanism by which MALAT1 regulates metastasis (Specific Aim 2). This project will lead to a major revision of the current model for a highly abundant and conserved lncRNA, and will profoundly advance the understanding of lncRNA's functions and mechanisms of action in tumor metastasis.

尽管在打击癌症方面取得了巨大进步，尤其是在早期阶段，转移仍然是一个艰巨而经常致命的挑战。显而易见的是，非编码RNA，包括 microRNA和长的非编码RNA（LNCRNA）是调节转移的分子网络的组成部分。一些LNCRNA与其基因组基因座具有相反的功能。例如，相反的表型已经从lncrna Haunt基因缺失和插入灭活中报道了，有趣的是，困扰基因缺失效应是由于基因组DNA的丧失，而不是丧失了Haunt RNA。因此，一个专业 lncRNA研究中的挑战是删除或灭活lncRNA基因引起的表型是否可以明确地归因于lncRNA本身的丢失或重叠调节的损失元素。 MALAT1（转移相关的肺腺癌转录本1）是最丰富的，并且正常组织中的保守LNCRNA，以前已被描述为转移启动子。然而，没有证据表明先前报道的malat1基因缺失（这导致了多个上调 MALAT1的相邻基因）或反义RNA（从未通过救援实验验证或通过 Malat1基因敲除细胞）效应特异性是malat1 lncRNA丢失。出乎意料的是，使用转录终止插入策略，我们发现破坏Malat1基因而不改变其表达乳腺癌的转基因小鼠模型中的相邻基因急剧促进了肺转移和重要的是，这种表型完全被malat1的遗传添加反转。此外，crispr-cas9- 在人乳腺癌细胞中介导的Malat1敲除诱导其转移能力，这被逆转由malat1重新表达。相反，MALAT1的过表达抑制了两者的乳腺癌转移转基因小鼠和异种移植模型。从机械上讲，我们使用了最近开发的染色质分离。 RNA纯化 - 质谱法（CHIRP-MS）方法，以识别Tead家族成员为结合蛋白对于原发性乳腺肿瘤组织的内源水平上的Malat1，MALAT1结合，隔离器，并灭活促旋转转录因子tead。我们还发现了一个反相关乳腺癌进展和转移的Malat1水平。基于这些重要发现，我们建议全面地表征Malat1在乳房中的功能丧失和功能障碍癌症转移，使用基因工程的小鼠模型，移植模型，合成模型，异种移植模型和CRISPR-CAS9基因组编辑方法（特定目标1）；我们还将阐明 MALAT1调节转移的机制（特定目的2）。该项目将导致对高度丰富且保守的LNCRNA的当前模型，并将深刻促进理解 lncRNA在肿瘤转移中的作用和作用机制。

项目成果

期刊论文数量（32）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

ATM-mediated stabilization of ZEB1 promotes DNA damage response and radioresistance through CHK1.

DOI：
10.1038/ncb3013
发表时间：
2014-09
期刊：
Nature cell biology
影响因子：
21.3
作者：
通讯作者：

Long noncoding RNA MALAT1 suppresses breast cancer metastasis.

DOI：
10.1038/s41588-018-0252-3
发表时间：
2018-12
期刊：
Nature genetics
影响因子：
30.8
作者：
Kim J;Piao HL;Kim BJ;Yao F;Han Z;Wang Y;Xiao Z;Siverly AN;Lawhon SE;Ton BN;Lee H;Zhou Z;Gan B;Nakagawa S;Ellis MJ;Liang H;Hung MC;You MJ;Sun Y;Ma L
通讯作者：
Ma L

Monoubiquitination in Homeostasis and Cancer.

稳态和癌症中的单泛素化