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.

项目摘要间充质干细胞（MSC）可以发展成成骨细胞，脂肪细胞和软骨细胞，从而提供再生医学材料。特别是，MSC的骨相关应用是最有前途的 MSC的临床应用。 PI最近发现FKBP4是FK506结合蛋白（FKBP）的成员肽酰丙酰丙酰基异构酶（PPIASE）家族促进MSC分化成骨细胞。他们还发现 FKBP4与Mettl3复合物相互作用，该复合物诱导了新型RNA修饰称为N6-甲基读（M6A）。尽管已知M6A参与MSC分化，但确切的作用和机制在很大程度上仍然存在未知。通过全基因组方法，PI在MSC中发现了数千个由M6A修饰的mRNA，成骨细胞和脂肪细胞。 mRNA还包括用于分化的关键转录因子基因作为几种修饰酶基因的组蛋白。此外，他们发现FKBP4激活了Mettl3复合物 PPIASE域依赖性方式。基于这些发现，PI假设FKBP4激活了在成骨细胞分化过程中其亚基之一的异构化通过异构化。他们也假设 M6A修饰通过调节RNA代谢而促进成骨细胞分化蛋白质水平提高了基因。 PI将以以下三个目标来检验这些假设。在AIM 1中， PI将在单个核苷酸的MSC，成骨细胞和脂肪细胞的转录组中绘制M6A分布。等级。随后，它们将以序列特异性的方式抑制甲基化 M6a和RNA代谢之间的关系。 AIM 2将研究FKBP4敲除的骨骼表型小鼠，还研究成骨细胞基因的M6a如何影响其分化。 AIM 3将研究M6A修改 FKBP4 mRNA作为FKBP4和METTL3之间的反馈。此外，此目标将研究FKBP4 在糖皮质激素受体分化过程中，抑制表达。这些研究总的来说展示了FKBP4的METTL3的新型调节机制，以及M6A修改如何控制MSC 分化。这些发现有望促进基于MSC的再生医学。