Novel Sources of Transplantable Beta-cell Replacements

描述（由申请人提供）：可以设想几种不同的细胞来源，用于产生用于治疗糖尿病的胰岛素产生细胞β-细胞。这些包括多能干细胞以及成体内胚层衍生物，如肝细胞。目前从多能前体产生β细胞的焦点是使用外源因子，而从肝细胞产生β细胞涉及使用基因转移策略的遗传重编程。该项目汇集了研究这些不同方法的研究人员，目的是整合相关信息，并利用这些信息指导我们寻找生产可移植β细胞等效物的最有效方法。具体而言，所有项目的分化中间体的表观遗传分析将用于为每个系统所需的外在/遗传操作的合理决策提供信息，以实现治疗有用细胞的最终目标。由于肝细胞的体内重编程需要使用腺病毒载体，因此必须阐明这种病毒在重编程中的作用。在每个系统中，预期体外产物的异质性，因此需要用于纯化表观遗传学上更均一的群体的试剂。为此，我们建议进一步开发表面反应性单克隆抗体。最后，分化产物的功能验证需要在移植到动物模型中时进行体内测试。 公共卫生相关性：从尸体供体移植胰岛已经表明，1型糖尿病可以成功地通过细胞疗法治疗。然而，高质量的尸体胰岛供体是罕见的，必须找到新的可移植β细胞来源，以使这种方法实现其临床潜力。该项目将产生患者匹配的自体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.