VCRFBI诱导脑出血反应性星形胶质细胞重编程为功能性神经元的研究

脑出血导致大量神经元损伤，同时胶质细胞增殖活化形成胶质瘢痕，阻碍神经再生，这是改善患者神经功能缺损的两大难点。很多研究通过移植干细胞替代损伤的神经元，或通过抑制胶质细胞增殖等方式改善神经再生的微环境，但疗效有限。我们前期研究发现，在体外脑出血细胞模型中，适宜浓度组合的六种简单易得、无免疫原性、组织渗透性强的小分子化合物（VCRFBI）能成功诱导大鼠反应性星形胶质细胞（RA）直接重编程为神经元。本项目拟利用小分子化合物介导的体细胞重编程技术同时解决这两大难点。因此我们设想：利用VCRFBI改变脑出血后RA的表观遗传特性，诱导RA直接重编程为功能性神经元，从而减轻胶质瘢痕，补充损伤的神经元，改善神经损伤。本项目拟在体外脑出血细胞模型、猕猴基底节区脑出血模型中验证上述假说，并利用显微光学切片断层成像（MOST）等技术三维可视化检测功能性神经再生。为临床治疗脑出血提供一种安全、有效的方法。

There are two major difficulties in improving the neurological deficits after intracerebral hemorrhage(ICH). On one hand, there is massive damage and loss of neurons which are difficult to achieve self-repair after injury. On the other hand, microenvironment such as glial scar resulted from activation and proliferation of astrocytes is not conducive to nerve regeneration. Aiming at these two difficulties, the existing researches either replace damaged neurons by transplanting stem cells, or improve the microenvironment of neural regeneration by inhibiting astrocyte proliferation, but only have limited efficacy. We have already found that six easily available、non-immunogenic、highly permeable to tissues small molecule compounds (VCRFBI) in appropriate concentration, can truely induce mouse reactive astrocytes'(RA) direct reprogramming to neurons in the in vitro ICH model. This project intends to solve these two difficulties at the same time using small molecule compounds-mediated somatic reprogramming technology. We envisage, using VCRFBI to change the epigenetic characteristics of reactive astrocytes (RA) after intracerebral hemorrhage, and induce RA's direct reprogramming to functional neurons, thereby reducing glial scars, replenishing damaged neurons, and improving neurological deficits after ICH. We intend to verify the above hypotheses in an in vitro ICH model and a basal ganglia cerebral hemorrhage model in cynomolgus monkeys, and visualize the recovery of nerve connections around the hemorrhage in an three-dimensional way using Micro-Optic Slice Tomography (MOST) technology. Finally, we will provide a safe and effective treatment for neural regeneration after clinical intracerebral hemorrhage.