类脑移植对脑缺血后血管新生的作用及机制

脑卒中是中国第一致死及主要致残病因，但缺乏有效的治疗手段。三维类脑具有人源化、细胞数量丰富、移植后存活率高等特点，有望发展成为脑卒中救治新策略。我们前期研究已证明类脑移植对脑缺血治疗有效，通过多重途径促进神经修复与再生，但对脑缺血后血管调控作用尚不清楚。因此，本项目拟采用同步辐射、基因编辑等技术，首次阐明类脑移植对脑缺血后血管新生的作用及机制：基于预实验发现类脑移植促进脑缺血后血管新生，进一步明确新生血管形态、功能与三维成像；探究移植的类脑中是否有细胞转分化为血管细胞、参与血管形成；最后基于预实验发现类脑可分泌高表达量的促血管生成经典因子HGF，制备HGF特异性敲除和过表达类脑，阐明类脑移植后通过分泌蛋白HGF调控脑缺血后血管新生的作用及机制。该项目有望证明类脑作为脑缺血潜在生物药的优越性(兼具神经和血管修复)，推动未来发展靶向类脑分泌蛋白的干预策略，为脑卒中类脑移植转化医学研究奠定基础。

Stroke is the first leading cause of death and main cause of disability in China, but with few effective therapies. With characteristics of humanization, large number of cells, high cell survival rate after transplantation and so on, the three-dimensional cerebral organoid (CO) shows the possibility to develop into a novel treatment strategy for stroke. Our previous study demonstrated that CO transplantation is an effective treatment for ischemic stroke, which promotes neural repair and regeneration of damaged brain tissue though multiple approaches. However, the effect of CO transplantation on angiogenesis after ischemic stroke is still unclear. Therefore, with the method of synchrotron radiation, CRISPR-Cas9 gene-editing system and others, this project plans to explore the effect and mechanism of CO transplantation on angiogenesis in ischemic stroke for the first time. According to preliminary experiment found that CO transplantation promotes angiogenesis after ischemic stroke, the project will further verify the morphology, function and three-dimensional imaging of new formed vessels, and examine the trans-differentiation of cells from transplanted CO to confirm the cell source of new formed vessels. And also, according to preliminary experiment found that CO can secrete high expression of classical angiogenic factor hepatocyte growth factor (HGF)，the project will prepare CO with HGF knockout and overexpression respectively, and elucidate the role and mechanism of CO secreted protein HGF on angiogenesis in ischemic stroke. The project hopes to prove the superiority of CO transplantation as a potential biological drug for ischemic stroke (with effect of neural repair and vascular repair), promote future development of novel intervention strategy targeting CO secreted protein, and further accumulate research foundation for the translational medicine of CO transplantation in stroke.