RNA methylation and mesenchymal stem cell differentiation

RNA甲基化与间充质干细胞分化

基本信息

批准号：
10549380
负责人：
Nobuaki Kikyo
金额：
$ 30.3万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-15 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10549380
关键词：
3&apos Untranslated Regions Acceleration Adipocytes Affect Alternative Splicing Biological Biological Assay Biology Cell Differentiation process Cell Proliferation Cells Chondrocytes Clinical Trials Complex Consensus Sequence Dexamethasone Differentiated Gene Enzymes Feedback Genes Genetic Transcription Glucocorticoid Receptor Histone Acetylation Histones In Vitro Isomerase Isomerism Knockout Mice Maps Mediating Mesenchymal Differentiation Mesenchymal Stem Cells Messenger RNA Methylation Methyltransferase Modeling Modification Molecular Molecular Analysis Molecular Conformation Mus Nuclear Export Nucleotides Osteoblasts Pathway interactions Peptides Peptidylprolyl Isomerase Phenotype Population Post-Transcriptional Regulation Proline Protein Family Proteins RNA RNA metabolism RNA methylation Regenerative Medicine Regulation Role Runx2 protein Site Tacrolimus Binding Proteins Techniques Terminator Codon Testing Translations Untranslated RNA adipocyte differentiation bone cell type clinical application demethylation genome-wide histone methylation immunoregulation innovation invention member novel osteoblast differentiation osteogenic protein folding protein function stem cell differentiation stem cell population stemness transcription factor transcriptome translational medicine virtual

项目摘要

Project Summary Mesenchymal stem cells (MSCs) can develop into osteoblasts, adipocytes, and chondrocytes, providing materials for regenerative medicine. In particular, bone-related applications of MSCs is one of the most promising clinical applications of MSCs. The PI recently found that Fkbp4, a member of the FK506-binding protein (Fkbp) family of peptidyl prolyl isomerase (PPIase), promotes MSC differentiation into osteoblasts. They also found that Fkbp4 interacts with the Mettl3 complex, which induces the novel RNA modification called N6-methyladenosine (m6A). Although m6A is known to be involved in MSC differentiation, exact roles and mechanisms remain largely unknown. Through a genome-wide approach, PI found thousands of mRNAs modified by m6A in MSCs, osteoblasts, and adipocytes. The mRNAs included critical transcription factor genes for the differentiation as well as several histone modifying enzyme genes. In addition, they found that Fkbp4 activates the Mettl3 complex in a PPIase domain-dependent manner. Based on these findings, the PI hypothesized that Fkbp4 activates the Mettl3 complex by isomerization of one of its subunits during osteoblast differentiation. They also hypothesized that m6A modifications promote osteoblast differentiation by modulating RNA metabolism with a result of increased protein levels of the genes. The PI will test these hypotheses with the following three aims. In Aim 1, the PI will map m6A distributions in the transcriptome of MSCs, osteoblasts, and adipocytes at a single nucleotide level. Subsequently, they will inhibit the methylation in a sequence-specific manner to understand causal relationships between m6A and RNA metabolism. Aim 2 will investigate bone phenotypes of Fkbp4 knockout mice and also study how m6A of osteoblast genes affect their differentiation. Aim 3 will study m6A modification of Fkbp4 mRNA as a feedback between Fkbp4 and Mettl3. In addition, this aim will investigate how Fkbp4 expression is inhibited during adipocyte differentiation by glucocorticoid receptor. Collectively, these studies will demonstrate a novel regulatory mechanism of Mettl3 by Fkbp4 and how m6A modifications controls MSC differentiation. These findings are expected to promote MSC-based regenerative medicine.

项目概要间充质干细胞（MSC）可以发育成成骨细胞、脂肪细胞和软骨细胞，提供再生医学材料。特别是，间充质干细胞的骨相关应用是最有前途的应用之一间充质干细胞的临床应用。 PI 最近发现 Fkbp4，FK506 结合蛋白 (Fkbp) 的成员肽基脯氨酰异构酶 (PPIase) 家族，促进 MSC 分化为成骨细胞。他们还发现 Fkbp4 与 Mettl3 复合物相互作用，诱导称为 N6-甲基腺苷的新型 RNA 修饰 (m6A)。尽管已知 m6A 参与 MSC 分化，但确切的作用和机制仍然很大程度上未知。通过全基因组方法，PI 在 MSC 中发现了数千个被 m6A 修饰的 mRNA，成骨细胞和脂肪细胞。 mRNA 还包括分化的关键转录因子基因作为几个组蛋白修饰酶基因。此外，他们发现 Fkbp4 激活 Mettl3 复合物 PPIase 域依赖性方式。基于这些发现，PI 假设 Fkbp4 激活 Mettl3 在成骨细胞分化过程中通过其亚基之一的异构化而形成复合物。他们还假设 m6A 修饰通过调节 RNA 代谢促进成骨细胞分化，结果基因的蛋白质水平增加。 PI 将通过以下三个目标来测试这些假设。在目标 1 中， PI 将绘制 MSC、成骨细胞和脂肪细胞转录组中单个核苷酸的 m6A 分布图等级。随后，他们将以序列特异性的方式抑制甲基化，以了解因果关系 m6A 和 RNA 代谢之间的关系。目标 2 将研究 Fkbp4 敲除的骨表型小鼠并研究成骨细胞基因的 m6A 如何影响其分化。 Aim 3将研究m6A修饰 Fkbp4 mRNA 作为 Fkbp4 和 Mettl3 之间的反馈。此外，该目标将研究 Fkbp4 如何在脂肪细胞分化过程中，糖皮质激素受体的表达受到抑制。总的来说，这些研究将展示 Fkbp4 对 Mettl3 的新型调控机制以及 m6A 修饰如何控制 MSC 差异化。这些发现有望促进基于间充质干细胞的再生医学。