MicroRNAs Regulating Gene Expression during Cellular Senescence and Aging

MicroRNA 在细胞衰老过程中调节基因表达

• 批准号: 8552404
• 负责人: Myriam Gorospe
• 金额: $ 35.46万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8552404
• 关键词: AUF1A protein; Affect; Aging; Aging-Related Process; Autophagocytosis; Biotin; Book Chapters; Cell Aging; Cell Cycle; Cells; Collection; Diploidy; Fibroblasts; Funding; Gene Expression; Gene Expression Profile; Human; Measures; Medicine; Messenger RNA; Metastatic malignant neoplasm to brain; Methods; MicroRNAs; Organism; Peroxisome Proliferator-Activated Receptors; Phenotype; Play; Polyribosomes; Process; Production; Proteins; Relative (related person); Reporter; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Stimulus; Stress; Translating; Translations; Untranslated RNA; age related; insight; interest; lipid biosynthesis; mRNA tagging; overexpression; programs; senescence; AUF1A protein; Affect; Aging; Aging-Related Process; Autophagocytosis; Biotin; Book Chapters; Cell Aging; Cell Cycle; Cells; Collection; Diploidy; Fibroblasts; Funding; Gene Expression; Gene Expression Profile; Human; Measures; Medicine; Messenger RNA; Metastatic malignant neoplasm to brain; Methods; MicroRNAs; Organism; Peroxisome Proliferator-Activated Receptors; Phenotype; Play; Polyribosomes; Process; Production; Proteins; Relative (related person); Reporter; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Stimulus; Stress; Translating; Translations; Untranslated RNA; age related; insight; interest; lipid biosynthesis; mRNA tagging; overexpression; programs; senescence

During aging, organisms show altered gene expression patterns and have an increasingly impaired ability to respond to stress-causing and mitogenic stimuli. Since post-transcriptional processes critically regulate changes in the collections of expressed proteins, the role of RBPs (described in other projects) and noncoding RNAs (particularly microRNAs and lncRNAs) are emerging as major factors controlling age-related gene expression patterns. To investigate ncRNA function during senescence, we employ approaches such as ncRNA reduction (by transfecting an antisense molecules), ncRNA overexpression (by transfecting precursors or mature ncRNA molecules), and identification of ncRNA-associated mRNAs by tagging the ncRNAs (using biotin or MS2 tags) and identifying target mRNAs through various methods (eg, microarray, RT-PCR). We investigate whether ncRNAs affect the stability of target mRNAs during senescence by measuring the steady-state levels and half-lives of the mRNAs of interest as a function of ncRNA abundance. We investigate whether ncRNAs affect the translation of target mRNAs by modulating ncRNA levels, and subsequently studying the relative association of the mRNA with translating polysomes and by quantifying the nascent translation rates of the encoded proteins. We also employ reporter constructs to gain additional insight into the processes modulated by ncRNAs and use different senescence-associated markers to examine changes in the senescence phenotype. During the past funding period, we have reported that the senescence-associated microRNA miR-519 plays a central role in autophagy (Abdelmohsen et al., Mol. Cell Biol, 2012), that miR-130 suppresses adipogenesis by lowering PPAR production (Lee et al., Mol. Cell Biol, 2011), and that miR-146 inhibits brain metastases (Hwang et al., Molecules and Cells, 2012). We have also shown that expression of a senescence-upregulated lncRNA (lincRNA-p21) is inhibited by the microRNA let-7 (Yoon et al., Mol. Cell 2012) and that a general factor in the production of microRNAs (Drosha) is inhibited by the senescence-downregulated protein AUF1 (Abdelmohsen et al., 2012). We reviewed the contributions of microRNAs in senescence and aging in several articles (Srikantan et al., Cell Cycle, 2011; Abdelmohsen and Gorospe, 2011; Srikantan et al., Cell Cycle, 2011) as well as in one book chapter (Grammatikakis and Gorospe, in MicroRNAs in Medicine 2012).

在衰老过程中，生物体显示出改变的基因表达模式，并且对应激和有丝分裂刺激的反应能力越来越受损。由于转录后过程严格调节表达蛋白的收集的变化，因此RBP（在其他项目中描述）和非编码RNA（尤其是microRNA和LNCRNA）的作用是控制年龄相关基因表达模式的主要因素。为了研究衰老过程中的NCRNA功能，我们采用了诸如NCRNA还原（通过转染反义分子），NCRNA过表达（通过转染前体或成熟的NCRNA分子）以及通过对NCRNA相关的MIRNA（使用NCRNAS）（使用生物2 tags）（使用生物2 tags）（使用生物2 tags）（通过识别NCRNA相关的MRNA）等方法（通过转染前体或成熟的NCRNA分子）（通过识别） RT-PCR）。我们研究了NCRNA是否通过测量感兴趣的mRNA的稳态水平和半衰期作为NCRNA丰度的函数来影响目标mRNA的稳定性。我们研究了NCRNA是否通过调节NCRNA水平来影响靶mRNA的翻译，然后研究mRNA与翻译多粒子体的相对关联并通过量化编码蛋白的新生翻译速率。我们还采用了记者构造来获得对NCRNA调节过程的更多见解，并使用不同的衰老相关标记来检查衰老表型的变化。 During the past funding period, we have reported that the senescence-associated microRNA miR-519 plays a central role in autophagy (Abdelmohsen et al., Mol. Cell Biol, 2012), that miR-130 suppresses adipogenesis by lowering PPAR production (Lee et al., Mol. Cell Biol, 2011), and that miR-146 inhibits brain metastases (Hwang et al., Molecules and细胞，2012年）。 我们还表明，衰老的lncRNA（Lincrna-p21）的表达受到microRNA let-7（Yoon等，Mol。Cell2012）的抑制，并且衰老蛋白质蛋白AUF1（Abdelmohsen等人，2012年）抑制了MicroRNAS（DROSHA）的一般因素（DROSHA）。我们回顾了microRNA在几篇文章中的衰老和衰老中的贡献（Srikantan等，Cell Cycer，2011； Abdelmohsen和Gorospe，2011; Srikantan等，Srikantan等，Cell Cycer，2011），以及一本书（Grammamatikakikis and Gorospe，Microsornas in Medicic in Medicine in Medicine in Medicate in Medicate in Medicate in Medicate in Medication in 2012）。