Identifying micro RNA regulators of senescence

基于衰老细胞中的生物信息学分析和miRNA筛选，我们已经鉴定了miR-340- 5 p诱导并潜在地调节细胞衰老。 以前，miR-340- 5 p被描述为肿瘤抑制miRNA，并作为抗病毒机制被耗尽。 然而，在这项研究中，我们提出了miR-340- 5 p在衰老中的新作用。 为了研究miR-340- 5 p参与细胞衰老的机制，我们将鉴定miR-340- 5 p的mRNA靶点、基因表达（RNA和蛋白质）的变化以及miR-340- 5 p水平改变的细胞中的代谢（目标1）。 我们将进一步研究miR-340- 5 p在细胞对衰老清除药物如达沙替尼和槲皮素组合的敏感性中的作用（目的2）。 最后，我们将在人类血液中检查miR-340- 5 p驱动的衰老蛋白和代谢物是否作为年龄的函数差异存在（目标3）。 其中几项研究将作为供资期结束后继续开展进一步研究的基础。 目前的努力 为了实现本研究的目标，我们已经开始实验性地解决这些问题： miR-340- 5 p的mRNA靶点是什么？ miR-340- 5 p调节导致的蛋白表达模式的变化是什么？ miR-340- 5 p对细胞代谢组学有什么影响？ miR-340- 5 p是否改变WI-38成纤维细胞对衰老清除化合物的敏感性？ miR-340- 5 p在人体组织中的丰度是否随衰老而变化？

Based on bioinformatic analysis and miRNA screening in senescent cells, we have identified miR-340-5p to induce and potentially regulate cellular senescence. Previously, miR-340-5p was described as a tumor-suppressing miRNA and is depleted as an antiviral mechanism. However, in this study, we suggest a novel role for miR-340-5p in senescence. To investigate the mechanisms by which miR-340-5p participates in cellular senescence, we will identify mRNA targets of miR-340-5p, changes in gene expression (RNA and protein), and metabolism in cells with altered miR-340-5p levels (Objective 1). We will further examine the role of miR-340-5p in cellular sensitivity to senolytic drugs like Dasatinib and Quercetin combination (Objective 2). Finally we will examine in human blood whether miR-340-5p-driven senescence proteins and metabolites are differentially present as a function of age (Objective 3). Several of these studies will serve as the basis for further studies to be continued beyond this funding period. CURRENT EFFORTS To accomplish the objectives of this study, we have embarked on addressing these questions experimentally: what are the mRNA targets of miR-340-5p? what are the changes in protein expression patterns as a result of miR-340-5p modulation? what are the effects of miR-340-5p on cell metabolomics? does miR-340-5p alter the sensitivity of WI-38 fibroblasts to senolytic compounds? does miR-340-5p abundance change in human tissues with aging?