Creating a Localized Pool of iPSCs In Vivo

 描述：慢性和退行性疾病会导致器官或组织随着时间的推移而恶化，是美国死亡和残疾的主要原因。干细胞具有恢复受损组织功能的能力，已被广泛应用于各种临床前模型以促进组织修复。然而，这些疗法的临床翻译由于与细胞分离、体外操作和移植相关的问题而变得复杂。体内诱导多能干细胞(IPSCs)的产生将绕过这些限制。研究表明，组织微环境可以支持体内的重新编程和转分化，但到目前为止还没有人尝试在特定的组织环境中在体内产生IPSCs。在这项研究中，我们将开发一种有效的方法来在体内重新编程细胞，使用微RNA簇miR302/367进行细胞重新编程和微环DNA进行基因传递。将讨论三个关键因素：1)体内重新编程的效率(AIM I)，2)体内产生的IPSCs的多能性状态(AIM II)，以及3)与重新编程相关的脱靶效应的记录(AIM III)。我们相信，在体内产生IPSCs是传统干细胞疗法的一个可行的替代方案。开发一种有效的组织特异性体内重编程方法为设计具有广泛应用范围的再生疗法奠定了必要的基础。

 DESCRIPTION: Chronic and degenerative diseases result in organ or tissue deterioration over time and are the leading cause of death and disability in the United States. Stem cells have the capacity to restore function to damaged tissues and have been used in a wide range of pre-clinical models to promote tissue repair. However, clinical translation of these therapies has been complicated by issues associated with cell isolation, ex vivo manipulation, and transplantation. In vivo generation of induced pluripotent stem cells (iPSCs) would circumvent these limitations. Studies have demonstrated that tissue microenvironments can support in vivo reprogramming and transdifferentiation, but in vivo generation of iPSCs within a specific tissue environment has not been attempted to date. In this study we will develop an efficient method for reprogramming cells in vivo using a combination of the micro RNA cluster miR302/367 for cellular reprogramming and minicircle DNA for gene delivery. Three key factors will be addressed: 1) efficiency of in vivo reprogramming (Aim I), 2) pluripotency state of the in vivo generated iPSCs (Aim II), and 3) documentation of off target effects associated with reprogramming (Aim III). We believe that generating iPSCs in vivo is a viable alternative to traditional stem cell therapies. Developing an efficient method for tissue specific in vivo reprogramming lays the necessary foundation for designing regenerative therapies with a broad range of applications.