人牙周膜干细胞成骨分化相关环状RNA的表达谱及circFAT1作用和机制的研究

项目来源

国家自然科学基金(NSFC)

项目主持人

郑云飞

项目受资助机构

北京大学

项目编号

81700938

立项年度

2017

立项时间

未公开

研究期限

未知 / 未知

项目级别

国家级

受资助金额

20.00万元

学科

医学科学-口腔颅颌面科学-口腔颅颌面组织器官缺损修复与再生

学科代码

H-H15-H1502

基金类别

青年科学基金项目

关键词

成骨分化 ; FAT1 ; 环状RNA ; 人牙周膜干细胞 ; hPDLSCs ; circRNA ; osteogenic differentiation ; circFAT1

参与者

贾凌飞;黄一平;李晓蓓;黄文雪;杨乔林;朱云艳

参与机构

首都医科大学

项目标书摘要:牙周炎和外伤等导致的牙周组织破坏是成年人失牙的主要原因,因而实现牙周组织再生是口腔医学研究的要务之一。人牙周膜干细胞(hPDLSCs)是牙周组织再生理想的种子细胞,其成骨分化能力是牙周组织再生的基础。非编码RNA对hPDLSCs成骨分化具有重要调控作用,但环状RNA(circRNA)在其间的作用尚无报道。本项目前期检测了hPDLSCs 成骨分化相关的 circRNA 表达谱,并针对特定的差异表达基因circFAT1进行了生物信息学研究,结果提示circFAT1可能通过结合 miR-181b、miR-34a、miR-3960 和 miR-376c,抑制hPDLSCs分化为成骨细胞。本研究拟通过体外细胞学、分子生物学和动物体内实验,进一步明确circFAT1对hPDLSCs成骨分化的作用,阐明其竞争性内源作用机制,试图以circFAT1为靶点促进hPDLSCs成骨分化以及牙周骨组织再生。

Application Abstract: The destruction of periodontal tissue caused by periodontal disease and trauma is the major cause of tooth loss in adults.Thus,to regenerate periodontal tissue is always highlighted in the field of stomatology.Human periodontal ligament stem cells(hPDLSCs)are the ideal seed cells for periodontal tissue regeneration,and the osteogenic potential of hPDLSCs is the basis of periodontal tissue repair and regeneration.Noncoding RNA has been shown to play an important role in regulating the osteogenic differentiation of hPDLSCs,however,the role of circular RNA(circRNA)was still unknown.In this study,we investigated the circRNA profiles during the osteogenic differentiation of hPDLSCs,and found that circRNA displayed stage-specific expression.Moreover,we performed bioinformatic analysis of circFAT1,one of the differentially expressed genes,and the results suggested that circFAT1 inhibited the osteogenic differentiation of hPDLSCs by binding miR-181b,miR-34a,miR-3960,and miR-376c.This study is conducive to determine the role of circFAT1 in regulating the osteogenic differentiation of hPDLSCs in vitro and in vivo,and to clarify the underlying competitive endogenous mechanism,shedding light on the enhancement of periodontal tissue regeneration by targeting circFAT1.

项目受资助省

北京市

项目结题报告(全文)

人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)具有多向分化潜能,是修复牙周组织较为理想的种子细胞。针对hPDLSCs成骨分化调控机制的研究有助于优化基于干细胞的牙周组织再生治疗。鉴于非编码RNA对间充质干细胞成骨分化的重要调控作用,本项目通过高通量测序的技术手段检测hPDLSCs成骨分化相关的全转录组,筛选了具有重要调控作用的circRNA和lncRNA,结果显示circFAT1、CDR1as、GAS5、MIR31HG、MEG3、FER1L4、MIR22HG等均在成骨细胞分化中具有潜在的调控作用。本项目进一步构建了circRNA-miRNA-mRNA以及lncRNA-mRNA的调控网络,并针对环状RNA circFAT1对hPDLSCs成骨分化的作用进行研究。体内和体外实验均显示敲低circFAT1能够促进hPDLSCs向成骨细胞分化,且circFAT1对NF-κB信号通路和miR-29b表达具有调控作用。以上研究结果对于全面认识非编码RNA在成骨细胞分化过程中的功能具有促进作用,完善了现有成骨细胞分化的调控网络,为临床利用干细胞进行牙周组织再生治疗提供依据。

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  • 1.Involvement of Noncoding RNAs in the Differentiation of Osteoclasts

    • 关键词:
    • NF-KAPPA-B; COLLAGEN-INDUCED ARTHRITIS; ACTIVATED PROTEIN-KINASE;GIANT-CELL TUMOR; BONE-RESORPTION; DOWN-REGULATION; REGULATESOSTEOCLASTOGENESIS; PROMOTES OSTEOCLASTOGENESIS; INHIBITSOSTEOCLASTOGENESIS; THERAPEUTIC TARGETS

    As the most important bone-resorbing cells, osteoclasts play fundamental roles in bone remodeling and skeletal health. Much effort has been focused on identifying the regulators of osteoclast metabolism. Noncoding RNAs (ncRNAs) reportedly regulate osteoclast formation, differentiation, survival, and bone-resorbing activity to participate in bone physiology and pathology. The present review intends to provide a general framework for how ncRNAs and their targets regulate osteoclast differentiation and the important events of osteoclastogenesis they are involved in, including osteoclast precursor generation, early differentiation, mononuclear osteoclast fusion, and multinucleated osteoclast function and survival. This framework is beneficial for understanding bone biology and for identifying the potential biomarkers or therapeutic targets of bone diseases. The review also summarizes the results ofin vivoexperiments and classic experiment methods for osteoclast-related researches.

    ...
  • 2.Involvement of Noncoding RNAs in the Differentiation of Osteoclasts

    As the most important bone-resorbing cells, osteoclasts play fundamental roles in bone remodeling and skeletal health. Much effort has been focused on identifying the regulators of osteoclast metabolism. Noncoding RNAs (ncRNAs) reportedly regulate osteoclast formation, differentiation, survival, and bone-resorbing activity to participate in bone physiology and pathology. The present review intends to provide a general framework for how ncRNAs and their targets regulate osteoclast differentiation and the important events of osteoclastogenesis they are involved in, including osteoclast precursor generation, early differentiation, mononuclear osteoclast fusion, and multinucleated osteoclast function and survival. This framework is beneficial for understanding bone biology and for identifying the potential biomarkers or therapeutic targets of bone diseases. The review also summarizes the results ofin vivoexperiments and classic experiment methods for osteoclast-related researches.

    ...
  • 3.Long Noncoding RNAs: New Players in the Osteogenic Differentiation of Bone Marrow- and Adipose-Derived Mesenchymal Stem Cells

    • 关键词:
    • Long noncoding RNAs; Mesenchymal stem cells; Osteogenic differentiation;Epigenetics; BMSCs; ADSCs;PROMOTES OSTEOBLAST DIFFERENTIATION; COMPETING ENDOGENOUS RNA;UP-REGULATION; HUMAN-DISEASE; GENE-EXPRESSION; STROMAL CELLS; IN-VITRO;LNCRNA; CANCER; IDENTIFICATION

    Mesenchymal stem cells (MSCs) are an important population of multipotent stem cells that differentiate into multiple lineages and display great potential in bone regeneration and repair. Although the role of protein-coding genes in the osteogenic differentiation of MSCs has been extensively studied, the functions of noncoding RNAs in the osteogenic differentiation of MSCs are unclear. The recent application of next-generation sequencing to MSC transcriptomes has revealed that long noncoding RNAs (lncRNAs) are associated with the osteogenic differentiation of MSCs. LncRNAs are a class of non-coding transcripts of more than 200 nucleotides in length. Noncoding RNAs are thought to play a key role in osteoblast differentiation through various regulatory mechanisms including chromatin modification, transcription factor binding, competent endogenous mechanism, and other post-transcriptional mechanisms. Here, we review the roles of lncRNAs in the osteogenic differentiation of bone marrow- and adipose-derived stem cells and provide a theoretical foundation for future research.

    ...
  • 4.Salivary Exosomes: Emerging Roles in Systemic Disease

    • 关键词:
    • saliva; exosome; biomarker; proteomics; genomics;SQUAMOUS-CELL CARCINOMA; EXTRACELLULAR VESICLES; PROTEOMIC ANALYSIS;ORAL-CANCER; MEDIATED ENDOCYTOSIS; POTENTIAL BIOMARKERS; PROMISINGBIOMARKER; DENDRITIC CELLS; RNA; PROTEIN

    Saliva, which contains biological information, is considered a valuable diagnostic tool for local and systemic diseases and conditions because, similar to blood, it contains important molecules like DNA, RNA, and proteins. Exosomes are cell-derived vesicles 30-100 nm in diameter with substantial biological functions, including intracellular communication and signalling. These vesicles, which are present in bodily fluids, including saliva, are released upon fusion of multivesicular bodies (MVBs) with the cellular plasma membrane. Salivary diagnosis has notable advantages, which include noninvasiveness, ease of collection, absence of coagulation, and a similar content as plasma, as well as increased patient compliance compared to other diagnostic approaches. However, investigation of the roles of salivary exosomes is still in its early years. In this review, we first describe the characteristics of endocytosis and secretion of salivary exosomes, as well as database and bioinformatics analysis of exosomes. Then, we describe strategies for the isolation of exosomes from human saliva and the emerging role of salivary exosomes as potential biomarkers of oral and other systemic diseases. Given the ever-growing role of salivary exosomes, defining their functions and understanding their specific mechanisms will provide novel insights into possible applications of salivary exosomes in the diagnosis and treatment of systemic diseases.

    ...
  • 5.Bone remodeling induced by mechanical forces is regulated by miRNAs

    • 关键词:
    • MESENCHYMAL STEM-CELLS; PERIODONTAL-LIGAMENT CELLS; FLUID SHEAR-STRESS;INDUCED OSTEOGENIC DIFFERENTIATION; ORTHODONTIC TOOTH MOVEMENT;DENTOALVEOLAR DISTRACTION OSTEOGENESIS; RAPID CANINE RETRACTION;SHOCK-WAVE THERAPY; OSTEOBLAST DIFFERENTIATION; PROMOTES OSTEOGENESIS

    The relationship between mechanical force and alveolar bone remodeling is an important issue in orthodontics because tooth movement is dependent on the response of bone tissue to the mechanical force induced by the appliances used. Mechanical cyclical stretch (MCS), fluid shear stress (FSS), compression, and microgravity play different roles in the cell differentiation and proliferation involved in bone remodeling. However, the underlying mechanisms are unclear, particularly themolecular pathways regulated by non-coding RNAs (ncRNAs) that play essential roles in bone remodeling. Amongst the various ncRNAs, miRNAs act as post-transcriptional regulators that inhibit the expression of their target genes. miRNAs are considered key regulators of many biologic processes including bone remodeling. Here, we review the role of miRNAs in mechanical force-induced bone metabolism.

    ...
  • 6.Salivary Exosomes: Emerging Roles in Systemic Disease

    • 关键词:
    • saliva; exosome; biomarker; proteomics; genomics;SQUAMOUS-CELL CARCINOMA; EXTRACELLULAR VESICLES; PROTEOMIC ANALYSIS;ORAL-CANCER; MEDIATED ENDOCYTOSIS; POTENTIAL BIOMARKERS; PROMISINGBIOMARKER; DENDRITIC CELLS; RNA; PROTEIN

    Saliva, which contains biological information, is considered a valuable diagnostic tool for local and systemic diseases and conditions because, similar to blood, it contains important molecules like DNA, RNA, and proteins. Exosomes are cell-derived vesicles 30-100 nm in diameter with substantial biological functions, including intracellular communication and signalling. These vesicles, which are present in bodily fluids, including saliva, are released upon fusion of multivesicular bodies (MVBs) with the cellular plasma membrane. Salivary diagnosis has notable advantages, which include noninvasiveness, ease of collection, absence of coagulation, and a similar content as plasma, as well as increased patient compliance compared to other diagnostic approaches. However, investigation of the roles of salivary exosomes is still in its early years. In this review, we first describe the characteristics of endocytosis and secretion of salivary exosomes, as well as database and bioinformatics analysis of exosomes. Then, we describe strategies for the isolation of exosomes from human saliva and the emerging role of salivary exosomes as potential biomarkers of oral and other systemic diseases. Given the ever-growing role of salivary exosomes, defining their functions and understanding their specific mechanisms will provide novel insights into possible applications of salivary exosomes in the diagnosis and treatment of systemic diseases.

    ...
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