Generation of induced neurons by direct conversion of microglia and related cells

小胶质细胞是一种广泛分布于中枢神经系统并能不断自我再生的髓样细胞。小胶质细胞归巢损伤位点的自然行为可用于多灶性神经病理学和神经再生approaches.The拟议的研究旨在探索小胶质细胞作为直接谱系转化为神经元命运的源细胞的潜力的靶向治疗。基于耗尽的内源性小胶质细胞可以通过造血来源的细胞外源性地再增殖的发现，还研究了这种小胶质细胞样细胞和先前的造血细胞的直接神经元重编程潜力。这项研究计划的另一个目的是开发一种模型，用遗传修饰的细胞系统地进入中枢神经系统，并引起广泛的体内神经发生。此外，将评估这种方法在损伤部位的靶向神经再生中的应用。将对小胶质细胞和相关细胞进行遗传修饰，并在体外筛选合适且有效的转录因子用于神经元重编程。将采用免疫组织学和电生理学方法以及基因表达谱对所得细胞的分子和功能特性进行评价。在诱导型操纵子的控制下用鉴定的前神经转录因子转导的细胞将用于替换减少的小胶质细胞，并且将在体内诱导神经元转化。该方法将应用于急性脑损伤模型，为直接神经元重编程的机制提供新的见解，并提高对小胶质细胞的性质和体内诱导神经元细胞特性的理解。直接在体内细胞重编程可以证明作为一个有价值的转化技术的再生疗法的发展。

Microglia are myeloid cells with ubiquitous distribution throughout the central nervous system and constant self-regeneration. The natural behavior of microglia to home to injury sites could be used for targeted therapies in multifocal neurological pathologies and neuroregenerative approaches.The proposed study aims to explore the potential of microglia as a source cell for direct lineage conversion towards a neuronal fate. Based on the finding that depleted endogenous microglia can be exogenously repopulated by cells of hematopoietic origin, direct neuronal reprogramming potential of such microglia-like cells and preceding hematopoietic cells is investigated as well. A further aim of this research proposal is to develop a model to systemically access the central nervous system with genetically modified cells and provoke wide-spread in vivo neurogenesis. Additionally, application of this approach for targeted neuroregeneration at lesion sites will be evaluated.Microglia and related cells will be genetically modified and in vitro screening for suitable and effective transcription factors for neuronal reprogramming will be conducted. Resulting cells will be evaluated regarding molecular and functional properties employing immunohistological and electrophysiological methods as well as gene expression profiling. Cells transduced with identified proneural transcription factors under the control of an inducible operator will be used to replace diminished microglia and neuronal conversion will be induced in vivo. This approach will then be applied in a model of acute cerebral injury.This project will provide additional insight into the mechanisms of direct neuronal reprogramming as well as improve the understanding of the nature of microglia and characteristics of in vivo induced neuronal cells. Direct in vivo cellular reprogramming could proof as a valuable translational technique for the development of regenerative therapies.