MEG3 deletion drives lung tumorigenesis due to environmental nickel exposure

由于环境镍暴露，MEG3 缺失导致肺部肿瘤发生

基本信息

批准号：
9852426
负责人：
Max Costa
金额：
$ 27.75万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9852426
关键词：
Affect Automobile Driving Biological Biological Assay CRISPR/Cas technology Cancer Patient Carcinogens Cell Culture Techniques Cells Clinical Development Down-Regulation Epigenetic Process Epithelial Cells Event Exposure to Gene Expression Regulation Genes Goals Human In Vitro Individual Inhalation International Agency for Research on Cancer Knock-out Knockout Mice Knowledge Laboratory Study Lung Lung Neoplasms Malignant - descriptor Malignant neoplasm of lung Mediating Mediator of activation protein Modeling Molecular Mus Nickel Nude Mice Oncogenes Oncogenic Outcome Oxidative Stress Play Pollution Prevention therapy Prognostic Marker Reporting Research Role Signal Pathway Signal Transduction Structure of parenchyma of lung System Testing Time Tissues Transducers Tumor Suppressor Genes Tumorigenicity Untranslated RNA Up-Regulation Xenobiotics attenuation base bronchial epithelium c-myc Genes carcinogenicity cell transformation deep sequencing design epidemiology study experimental study gain of function improved in vivo insight knock-down loss of function lung carcinogenesis lung tumorigenesis mouse model new therapeutic target novel overexpression professional atmosphere small hairpin RNA therapeutic target

Affect Automobile Driving Biological Biological Assay CRISPR/Cas technology Cancer Patient Carcinogens Cell Culture Techniques Cells Clinical Development Down-Regulation Epigenetic Process Epithelial Cells Event Exposure to Gene Expression Regulation Genes Goals Human In Vitro Individual Inhalation International Agency for Research on Cancer Knock-out Knockout Mice Knowledge Laboratory Study Lung Lung Neoplasms Malignant - descriptor Malignant neoplasm of lung Mediating Mediator of activation protein Modeling Molecular Mus Nickel Nude Mice Oncogenes Oncogenic Outcome Oxidative Stress Play Pollution Prevention therapy Prognostic Marker Reporting Research Role Signal Pathway Signal Transduction Structure of parenchyma of lung System Testing Time Tissues Transducers Tumor Suppressor Genes Tumorigenicity Untranslated RNA Up-Regulation Xenobiotics attenuation base bronchial epithelium c-myc Genes carcinogenicity cell transformation deep sequencing design epidemiology study experimental study gain of function improved in vivo insight knock-down loss of function lung carcinogenesis lung tumorigenesis mouse model new therapeutic target novel overexpression professional atmosphere small hairpin RNA therapeutic target

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项目摘要

Abstract Decades of both epidemiological research and in vitro and in vivo laboratory studies have consistently reported an association between environmental nickel exposure and lung cancer. As early as 1990, the International Agency for Research on Cancer (IARC) classified nickel compounds as Group 1 carcinogens: substances confirmed as carcinogenic to humans. Although the multiple mechanisms, including oxidative stress, epigenetic effects, and activation of signaling pathways that trigger differential gene regulation has been thought to be associated with nickel carcinogenic effect, there is no evidence that a single nickel- inducible factor can drive transformation of human bronchial cells, to the best of our knowledge. In our Preliminary Studies, we performed long noncoding RNA (lncRNA) deep sequencing using the Illumina HiSeqTM2000/2500 high throughout system to evaluate whether nickel affects the abundance of 5,929 known lncRNAs. Nickel treatment altered the levels of 24 of these lncRNAs in normal primary human bronchial epithelial cells (HBECs), while 16 of them were verified in Real-time PCR assay. With overexpression and/or knockdown analyses of the 16 lncRNAs, we further discovered that a reduced abundance of maternally-expressed gene 3 (MEG3) was sufficient for malignant transformation of HBECs. Our demonstration that MEG3 inhibition can independently transform HBECs provides the basis of our central hypothesis that MEG3 downregulation drives transformation and tumorigenecity of HBECs after nickel exposure. Here we propose to elucidate the molecular mechanisms that underlie our novel findings that MEG3 downregulation is a crucial driver for nickel-induced malignant transformation of HBECs with the following Specific Aims: 1, To test the hypothesis that p62 and C-Myc define an important MEG3-regulated axis that promotes transformation of HBECs upon MEG3 deficiency; 2, To test the hypothesis that the crosstalk between a p62/C-Myc cascade and the PHLPP/HIF-1α axis causes the MEG3-deficiency- correlated malignant transformation of HBECs; 3, To explore the biological significance of MEG3 deletion and its activated downstream molecules in lung tumoriginecity. Our novel Preliminary Findings suggest that MEG3 is downregulated by nickel exposure both in vitro and in vivo, that knockdown of MEG3 alone can transform HBECs, and that crosstalk between the putative signaling transducers downstream of MEG3 mediate the malignant transformation of HBECs caused by MEG3 deficiency. This proposal's strengths are the complementary use of cell culture models and novel conditional MEG3 knockout mouse models to examine integration of the molecular events that account for nickel-mediated lung carcinogenesis. Clarifying these issues will provide valuable insights regarding MEG3 as a prognostic biomarker and/or as a therapeutic target. Ultimately, both uses could improve clinical outcomes in lung cancer patients.

抽象的流行病学研究以及体外和体内实验室研究的数十年始终如一报道了环境镍暴露与肺癌之间的关联。早在1990年，国际癌症研究机构（IARC）将镍化合物归类为第1组致癌物：对人类的致癌物质。虽然多种机制，包括氧化触发差异基因调节的压力，表观遗传效应和信号通路的激活已被认为与镍致癌作用有关，没有证据表明单个镍据我们所知，诱导因子可以驱动人支气管细胞的转化。在我们的初步研究，我们使用Illumina进行了长的非编码RNA（LNCRNA）深度测序 HISEQTM2000/2500在整个系统中高高，以评估镍是否影响5,929的抽象已知的lncrnas。镍治疗改变了正常原代人中24个LNCRNA的水平支气管上皮细胞（HBEC），其中16个在实时PCR测定中进行了验证。和 16个LNCRNA的过表达和/或敲低分析，我们进一步发现母体表达的基因3（MEG3）的抽象足以使HBEC的恶性转化。我们表明MEG3抑制可以独立改变HBEC的演示提供了我们的基础 MEG3下调驱动HBEC的转化和肿瘤性的中心假设镍暴露。在这里，我们建议阐明我们新发现的基础的分子机制 MEG3下调是镍引起的HBEC的恶性转化的关键驱动力以下特定目的：1，检验p62和c-myc定义重要的MEG3调节的假设在MEG3缺乏症上促进HBEC转换的轴； 2，检验以下假设 p62/c-myc级联与PHLPP/HIF-1α轴之间的串扰导致MEG3缺乏症 HBEC相关的恶性转化； 3，探索MEG3缺失的生物学意义并在肺肿瘤中激活的下游分子。我们的小说初步发现表明 MEG3在体外和体内都被镍暴露而下调，仅MEG3就可以转换HBEC，并在MEG3下游的假定信号传感器之间进行串扰调解由MEG3缺乏引起的HBEC的恶性转化。该提议的优势是细胞培养模型和新颖有条件的MEG3基因敲除鼠标模型的完整使用检查解释镍介导的肺癌发生的分子事件的整合。澄清这些问题将提供有关MEG3作为预后生物标志物和/或AS的宝贵见解治疗靶标。最终，两种用途都可以改善肺癌患者的临床结果。