HIF-1α诱导下LncRNA (TCONS_00072170)-miR143-KRAS调控内皮祖细胞参与牵张成骨血管形成的机制研究

牵张成骨（DO）成骨速率达婴幼儿生长发育速度的4至6倍，远超骨折愈合，其成骨机制复杂，目前尚无定论，成为当今国内外研究聚焦的热点之一。骨是高度血管化的组织，血管形成是新骨形成的先决条件。本课题组在前期DO成骨机制的序列研究中，发现DO组织内的血管快速形成，而缺氧微环境、内皮祖细胞和非编码RNA在其中起到举足轻重的作用。本课题以TCONS_00072170-miR143-KRAS信号轴为线索，利用RNA-Scope技术定位TCONS_00072170在组织或细胞中的位置，通过ChIP、RIP及RNA pull-down等研究核酸和蛋白互作的方法，构建非编码RNA及其靶基因敲除（CAS9介导）、沉默、过表达慢病毒质粒，结合体内外实验，探索HIF-1α诱导下以TCONS_00072170为中心调控DO中EPCs血管形成的DNA-RNA-蛋白质互作网络，为阐明DO快速成骨的生物学机制提供新的思路。

The osteogenesis rate of distraction osteogenesis (DO) is 4 to 6 times the growth rate of infants, far beyond fracture healing. The osteogenesis mechanism of DO is complicated and inconclusive, becoming one of the research hotspots at home and abroad. Bone is a highly vascularized tissue and neovasculogenesis serves as a prerequisite for new bone formation. In the sequence study on the pre-DO osteogenic mechanism, our team found rapid formation of blood vessels in the DO tissue, in which hypoxic microenvironment and EPC non-coding RNA played a pivotal role. Taking TCONS_00072170-miR143-KRAS signal axis as a clue, this project uses RNA-Scope technology to locate TCONS_00072170 in tissues or cells, constructs non-coding RNA and its target gene knockout (CAS9-mediated), silenced, over-expressed lentiviral plasmids through research methods on nucleic acid and protein interactions, such as ChIP, RIP, and RNA pull-down, and explores the DNA-RNA-protein interactions of TCONS_00072170-induced regulation of EPCs neovascularization in DO under HIF-1α combined with in vivo and in vitro experiments. The network will provide new ideas for clarifying the biological mechanism of DO rapid osteogenesis.