Generation and integration of new CNS neurons by in vivo directed conversion

描述（由申请人提供）：本申请的目的是研究使用原位定向神经元转化（IDNC）将成人CNS的现有体内分化细胞（如星形胶质细胞）转化为功能性和适当整合的神经元的可行性和要求。IDNC是一种新颖的方法，与干细胞方法不同。使用IDNC，使用多顺反子慢病毒载体或其它方式将转录调节剂的混合物转导到成熟CNS细胞，例如星形胶质细胞中。这些因子导致快速和定向转化为独特的成熟神经元表型。这种定向转化挑战了分化的细胞表型固有稳定的基本原则，特别是在细胞中原位稳定。 CNS。定向转化潜在地克服了其他治疗性细胞替代策略（例如干细胞方法）的一些固有困难。直接转换现有单元对现有电路的一般架构的干扰可能最小。第二个，也许是更大的挑战是新转换的神经元到中枢神经系统局部电路的功能整合。这个问题将使用各种生理和光学成像方法来解决。 公共卫生相关性：适当整合到成熟哺乳动物CNS中的新CNS神经元的产生代表了再生医学的主要挑战。我们提出了一种新的和概念上简单的方法：产生新的哺乳动物中枢神经系统神经元原位在体内直接转换的其他中枢神经系统细胞。我们将确定这些细胞是否可以整合到现有的神经元回路中。

DESCRIPTION (provided by applicant): The goal of this application is to investigate the feasibility and requirements of converting existing in vivo, differentiated cells of the adult CNS - such as astroglia - into functional and appropriately integrated neurons, using in situ directed neuronal conversion (IDNC). IDNC is novel and distinct from stem cell approaches. With IDNC, a cocktail of transcription regulators is transduced into mature CNS cells, such as astroglia, using polycistronic lentiviral vectors or otherwise. These factors lead to a rapid and directed conversion into a distinct mature neuronal phenotype. Such directed conversion challenges the basic tenet that differentiated cell phenotypes are inherently stable - particularly in situ in the CNS. Directed conversion potentially overcomes some of the inherent difficulties with other therapeutic cell replacement strategies, such as stem cell approaches. Directly converting existing cells may be least intrusive on the general architecture of existing circuitry. A second and perhaps greater challenge is the functional integration of newly converted neurons into CNS local circuitry. This problem will be address using a variety of physiological and optical imaging approaches. PUBLIC HEALTH RELEVANCE: The generation of new CNS neurons that appropriately integrate into the mature mammalian CNS represents a major challenge in regenerative medicine. We present a novel and conceptually simple approach: to generate new mammalian CNS neurons in situ by in vivo directed conversion of other CNS cells. We will determine whether such cells can integrate into the existing neuronal circuitry.