circPOMT1靶向hsa-miR-6881-3p调控人脂肪干细胞成骨向分化的作用机制

Maxillofacial bone defects affect the quality of life of patients seriously. Bone tissue engineering is expected to become a new way of bone defect repair. However, the specific mechanism of osteogenic differentiation of seed cells is still unclear, and how to effectively and safely regulate osteogenic differentiation of seed cells is one core issue in bone tissue engineering. . Our preliminary RNA high-throughput sequencing experiment showed that 210 circRNAs were related with the osteogenic differentiation of human adipose stem cells (hASCs). circPOMT1 was identified by qRT-PCR and RNA enzyme tolerance test. We found that circPOMT1 could inhibited the osteogenic differentiation of hASCs, while hsa-miR-6881-3p could promote the osteogenic differentiation of hASCs. There was one hsa-miR-6881-3p binding site in the base sequence of circPOMT1, and the expression value of hsa-miR-6881-3p was negatively correlated with circPOMT1. There were binding sites of Smad6 and Chordin in the base sequence of hsa-miR-6881-3p. Smad6 can inhibit the formation of R-Smad and co-Smad complex, and thus serves as an endogenous inhibitor of BMPs signaling pathway. Chordin can be bound to BMP ligands to prevent the ligand-receptor interaction. Therefore, it is speculated that circPOMT1 may target hsa-miR-6881-3p to affect the osteogenic differentiation of hASCs via BMPs signaling pathway. . On this basis, we intend to use series experiments in vitro and in vivo, and methods of mechanism study to clarify the interaction relationship and specific mechanism between circPOMT1 and hsa-miR-6881-3p, the effect and specific mechanism of circPOMT1–hsa-miR-6881-3p on the osteogenic differentiation of hASCs. . Through the implementation of this project, it is hopeful to elucidate the specific roles of the cross-talking of circPOMT1–hsa-miR-6881-3p in the osteogenic differentiation of hASCs, which will contribute to the safe and efficient repair of maxillofacial bone defects, and provide a novel way for maxillofacial bone defects repair.

颌面部骨缺损严重影响患者生活质量，骨组织工程有望成为骨缺损修复新模式。但如何高效、安全地调控种子细胞成骨分化是目前骨组织工程的一项核心问题。申请人前期研究表明，circPOMT1可抑制hASCs成骨分化，而miR-6881-3p可促进成骨分化；circPOMT1碱基序列中存在miR-6881-3p结合位点且两者表达呈负相关，miR-6881-3p存在BMPs信号抑制物Smad6和Chordin的结合位点。因此推测，circPOMT1可能通过靶向miR-6881-3p，调控BMPs信号，影响成骨分化。在此研究基础上，本项目拟通过体外及体内实验验证、作用机制探索的技术路线，明确circPOMT1–miR-6881-3p交互作用及成骨调控机制。本项目有望阐明circPOMT1–miR-6881-3p在成骨向分化中的功能及作用机制，为临床特异性地修复颌面骨缺损提供新思路。

颌面部骨缺损可造成患者颜面部塌陷、牙列缺损以及咬合紊乱等症状，破坏患者颜面美观，并造成患者咀嚼发音等口腔功能障碍，严重影响患者的身心健康和生活质量，如何安全高效地治疗颌面部骨缺损是目前口腔医学中的一大难题。基于hASCs再生修复的骨组织工程是目前治疗颌面部骨缺损的新兴技术。circRNAs是近年来被发现的一类广泛存在于真核细胞转录组的ncRNAs，作为理想的诊疗标记物，circRNAs对成骨相关信号通路有调控作用，本研究通过探究 circPOMT1-hsa-miR-6881-3p 调控 hASCs 成骨向分化的机制及相关分子靶点，明确了circPOMT1与hsa-miR-6881-3p的结合位点，以及hsa-miR-6881-3p与Smad6的结合位点，基本阐明circPOMT1与hsa-miR-6881-3p以及hsa-miR-6881-3p与Smad6的交互作用机制，并明确circPOMT1/hsa-miR-6881-3p/Smad6对hASCs成骨分化的作用，有望利用该机制精细调控 hASCs 成骨向分化。同时对miR-20a-5p调控成骨及成软骨分化的作用及机制进行了研究，基本明确了miR-20a-5p对骨软骨再生的调控作用，为临床颌面部骨缺损修复提供新思路新证据。