Novel Sources of Transplantable Beta-cell Replacements

描述(由申请人提供)：可以设想几种不同来源的细胞，用于产生胰岛素产生细胞，用于治疗糖尿病的β细胞。其中包括多能干细胞和成体内胚层衍生物，如肝细胞。目前从多能前体细胞产生β细胞的重点是使用外部因子，而从肝细胞产生β细胞涉及到使用基因转移策略的遗传重新编程。该项目将致力于这些不同方法的研究人员聚集在一起，目的是整合相关信息，并利用这些信息指导我们以最有效的方式生产可移植的β细胞等效物。具体地说，对所有项目的分化中间体的表观遗传学分析将被用于为理性决策提供信息，以了解每个系统为实现治疗有用细胞的最终目标所需的外在/遗传操作。由于肝细胞体内重编程需要使用腺病毒载体，因此必须阐明该病毒在重编程中的作用。体外产物的异质性在每个系统中都是预期的，因此需要试剂来纯化更多表观遗传均一的种群。为此，我们建议进一步开发表面反应性单抗。最后，分化产物的功能验证需要在移植到动物模型后进行体内测试。 公共卫生相关性：来自身体捐赠者的胰岛移植表明，通过细胞疗法可以成功地治疗1型糖尿病。然而，高质量的身体胰岛供体是罕见的，必须找到新的可移植的β细胞来源，以便这种方法实现其临床潜力。该项目将产生与患者匹配的自体β细胞，用于治疗1型糖尿病。成功的执行将影响世界各地的大量患者。

DESCRIPTION (provided by applicant): Several different sources of cells for the generation of insulin producing cells beta-cells for the treatment of diabetes can be envisioned. These include pluripotent stem cells as well as adult endoderm derivatives such as liver cells. The current focus in producing beta-cells from pluripotent precursors is on the use of extrinsic factors, whereas the generation of beta-cell from hepatic cells involves genetic reprogramming using gene transfer strategies. This project brings together investigators working on these different approaches for the purpose of integrating relevant information and using it to guide us towards the most efficient way of producing transplantable beta-cell equivalents. Specifically, epigenetic analysis of differentiation intermediates from all projects will be used to inform rational decisions about the extrinsic/genetic manipulations required in each system to achieve the final goal of therapeutically useful cells. Because in vivo reprogramming of liver cells requires the use of adenoviral vectors, the role of this virus in reprogramming must be elucidated. Heterogeneity of the in vitro products is expected in each system and hence reagents to purify more epigenetically homogeneous populations are needed. We propose further development of surface reactive monoclonal antibodies for this purpose. Finally, functional validation of the differentiation products requires in vivo testing upon transplantation into animal models. PUBLIC HEALTH RELEVANCE: Transplantation of islets from cadaveric donors has shown that type 1 diabetes can be successfully treated by cell therapy. However, high quality cadaveric islet donors are rare and new sources of transplantable beta-cells must be found in order for this approach to realize its clinical potential. This proejct will produce patient-matched autologous beta-cells for the treatment of type 1 diabetes. Successful execution will impact large numbers of patients world-wide.