血管内皮来源的模式识别受体NLRC5改善血管新生的机制及治疗研究

Cardiovascular disease is one of the commonest disease, which would risk the health and life of Chinese people. Angiogenesis plays an important role in alleviating and treating tissue damage and necrosis caused by ischemic disease, but the definite mechanism is still unknown. Previous studies have shown that the innate immune protein, NLRs family, play important roles in the inflammation as well as ischemic injury response. Our studies have found that NLRC5 protein in the NLRs family was involved in the pathological process of vascular diseases. Besides, it has found that NLRC5 is translocated into the nucleus of endothelial cells in vivo and in vitro and is involved in angiogenesis after the stimulation. In the model of femoral artery ligation, NLRC5 knockdown can significantly inhibit angiogenesis. In this project, we intend to explore the function of NLRC5 on angiogenesis in vivo by using femoral artery ligation, bone marrow transplantation and genetic lineage tracking. Western blot, CUT&Tag -sequence, protein spectrum analysis and immunoprecipitation will be used to investigate the effect and mechanism of NLRC5 on angiogenesis. By constructing plasmids of different domains, the main domains that NLRC5 plays a role in angiogenesis were discussed. This study will clarify the role and mechanism of NLRC5 on blood vessels and provide new ideas and targets for the treatment of diseases involved angiogenesis.

心血管疾病是危害国人健康的最主要疾病，血管新生对于治疗缺血性疾病所致的损伤和坏死意义重大，但机制仍然未完全阐明。以往研究显示模式识别受体NLRs家族主要调控炎症和缺血损伤反应，我们研究发现NLRs家族中的NLRC5蛋白参与血管重构的病理过程。前期研究发现体内、外的内皮细胞激活后，NLRC5蛋白进入内皮细胞核，并参与血管新生。在下肢缺血模型中，发现NLRC5敲除小鼠血管新生反应明显受到抑制。本项目中，拟采取下肢缺血模型以及骨髓移植和遗传谱系追踪方法探讨NLRC5对体内血管新生作用。机制上将采用Western blot、CUT&Tag-sequence、蛋白质谱、免疫共沉淀等技术探讨NLRC5对血管新生的作用和机制。并通过构建不同结构域的质粒，探讨NLRC5对血管新生起作用的主要分子结构域。通过本研究将明确NLRC5对血管新生的作用和机制，为血管新生治疗提供新的思路和靶点。

心血管疾病是危害国人健康的最主要疾病，血管新生对于治疗缺血性疾病所致的损伤和坏死意义重大。在本项目中，我们发现模式识别受体NLRs家族中的NLRC5蛋白参与血管重构的病理过程。该蛋白在体内、外的内皮细胞激活后，NLRC5蛋白进入内皮细胞核，并参与血管新生。在下肢缺血模型中，发现NLRC5敲除小鼠血管新生反应明显受到抑制。采取下肢缺血模型以及骨髓移植和遗传谱系追踪方法明确证明NLRC5可促进病理性血管新生。随后通过Western blot、RNA-seq、免疫共沉淀等技术发现并证实NLRC5入核后通过增强STAT3蛋白在细胞核的转录作用，进而发挥血管新生促进作用。并通过构建不同结构域的质粒，证实NLRC5的NACHT+DD结构域蛋白就能发挥血管新生起主要作用（该结构域已申请专利）。通过本研究将明确NLRC5对血管新生的作用和机制，为血管新生治疗提供新的思路和靶点。