Pancreatic Islet Design & Engineering (SysCODE 3 of 10)

内分泌胰腺既是发育生物学中基本问题的底物，也是 这种疾病的目标是糖尿病，它影响着全世界数百万人。详细的遗传基因 如果我们要操纵胰腺内分泌发育的分子机制将是必不可少的。 活体内胰岛细胞的命运和数量。我们对胰腺发育的新理解是， 通过与周围间充质的相互作用，最初未形成图案的胰腺上皮 先后细分为外分泌室和内分泌室，这些内分泌室随后分化， 后者包含产生胰岛素的|3eta细胞。在决定胰岛细胞规格的因素中有 许多基因的身份现在已为人所知。事实上，现在可以将这些基因排序为第一个 根据基因作用、表达和功能相互依赖的顺序对遗传调控网络进行排序， 和等级关系。因此，这笔赠款提出了一个问题：我们是否已经知道 足够了，我们能否充分增强这一知识，开始利用这些信息来系统地 在体外设计胰岛和胰岛细胞？ 为了实现这一雄心勃勃的目标，我们将实现三个具体目标，这些目标与 SysCODE的多个组件。在目标1中，我们将生成关键早期状态的完整基因列表 发育中的内分泌胰腺。这一领域的初步努力已经完成(Gu等人， 发展131、165-79、2004年)。我们现在将使用来自其他数据的数据来增强此信息 发育阶段、选定的突变状态和第一代蛋白质组学分析。在目标2中，在 与SysCODE计算团队合作，我们将开发方法来对这些基因和 选择的蛋白质以对两位生物学家都有用的形式进入最终的基因调控网络(CRN) 和组织工程师。最后，在目标3中，我们将与SysCODE组织工程团队合作 实施基于干细胞的胰岛发育工程模型，我们将使用GRN 在AIM 2中生成，以优化胰腺内分泌命运规范。在拨款的最后几年，我们将 将改造后的胰岛移植到糖尿病小鼠体内，并评估它们的生理功能。总而言之，这些 与SysCODE的其余部分一起努力，将为再生性建立一个转型范例 医药。

The endocrine pancreas is both the substrate for fundamental questions in developmental biology as well as the target of the disease diabetes mellitus, which affects millions of individuals worldwide. A detailed genetic and molecular understanding of pancreatic endocrine development will be essential if we are to manipulate islet cell fate and numbers in vivo. Our emerging understanding of pancreatic development is one in which, through interactions with surrounding mesenchyme, the initially unpatterned pancreatic epithelium is successively sub-divided into exocrine and endocrine compartments which subsequently differentiate, the latter containing the |3eta cells that produce insulin. Among the factors that dictate islet cell specification are many genes whose identities are now known. In fact, it is now possible to order these genes into a first order genetic regulatory network in terms of order of gene action, expression and functional interdependencies, and hierarchical relationships. This grant therefore poses the question: Do we already know enough, and can we sufficiently augment that knowledge, to begin to use this information to systematically engineer islets and islet cells in vitro? To accomplish this ambitious goal, we will undertake three Specific Aims that are highly integrated with multiple components of SysCODE. In Aim 1, we will generate complete gene lists for key early states in the developing endocrine pancreas. An initial effort in this area has already been accomplished (Gu et al., Development 131, 165-79, 2004). We will now augment this information with data from additional developmental stages, selected mutant states and first-generation proteomic analyses. In Aim 2, in conjunction with the SysCODE Computational Team, we will develop methodology to order these genes and selected proteins into a definitive gene regulatory network (CRN) in a format that is useful to both biologists and tissue engineers. Lastly, in Aim 3 we will collaborate with the SysCODE Tissue Engineering Team to implement a stem cell based, engineered model of pancreatic islet development and we will use the GRNs generated in Aim 2 to optimize pancreatic endocrine fate specification. In the out years of the grant, we will transplant the engineered islets into diabetic mice and assess their physiological function. Collectively, these efforts, in conjunction with the rest of SysCODE, will establish a transforming paradigm for regenerative medicine.