Genetic control of pancreatic endocrine cell development

描述(由申请者提供)：这个项目将使用基于团队的方法来追求三个直接相关的研究领域，这些领域对于学习从其他细胞类型制造新的胰岛β细胞非常重要。在目标1中，我们试图更深入地了解在胰腺发育过程中决定多种内分泌细胞类型形成的分子事件。这将通过分离一系列高度纯化的祖细胞群体，进行数字基因表达分析，并开发生物信息学策略来表征离散细胞群体之间的差异，这些差异将在时间上或遗传上提供信息。在目标2中，我们将寻求利用在目标1中获得的知识来确定用于指导hESCs向胰腺细胞命运分化的方案的准确性，并通过监测在小鼠发育过程中被特异性激活或抑制的基因簇的表达来合理地改进这些方案。在目标3中，我们将开发新的小鼠品系，在这些品系中，三种转录因子的表达被报道能够将胰腺腺泡转化为β细胞，这些转录因子的表达可以很容易地通过给予多西环素来调节。这些小鼠将被用来探索转分化实际上是如何发生的，并确定新产生的贝塔细胞具有多大的功能，并显示出类似于真正的贝塔细胞的基因表达谱。这个项目基于这样的前提/假设，即在小鼠发育过程中通常建立的多个基因调控网络必须在其他类型的细胞定向或转分化过程中建立，以实现临床使用所需的类似β细胞的功能。更多关于胰腺特异性基因调控网络的知识，它们是如何建立的，在相关细胞群体中是如何建立和不同的，以及确定它们是否存在于实验衍生的细胞群体中，将成为新发现和方案改进的平台。该项目中的六名研究人员拥有卓有成效的协作互动的记录，并带来了实现这些目标所需的特定知识和能力。因此，预计该项目将产生重要的资源，不仅将促进Beta细胞生物学联合会的两个总体目标，而且还将产生广泛的科学影响和实用价值。 公共卫生相关性：1型和2型糖尿病是导致严重发病率和死亡率的疾病，因此具有不利的经济影响。这两种疾病的特征都是分泌胰岛素的胰岛β细胞遭到破坏或功能障碍。这项申请寻求获得关键信息，以开发新的、基于细胞的替代疗法，这些疗法有望实现比目前可能的更好的血糖控制。

DESCRIPTION (provided by applicant): This project will use a team-based approach to pursue three directly interrelated areas of investigation important for learning to make new pancreatic beta cells from other cell types. In Aim 1 we seek to gain a deeper understanding of the molecular events that dictate formation of multiple endocrine cell types during pancreas development. This will be achieved by isolating a series of highly purified progenitor cell populations, performing digital gene expression analysis, and developing bioinformatics strategies for characterizing the differences among discrete cellular populations that will be temporally or genetically informative. In Aim 2 we will seek to utilize the knowledge gained in Aim 1 to determine the veracity of protocols being used to direct the differentiation of hESCs towards pancreatic cell fates and to rationally improve these protocols by monitoring the expression of gene clusters that are specifically activated or repressed during mouse development. In Aim 3 we will develop new mouse lines in which the expression of three transcription factors that have been reported to be capable of transdifferentiating pancreatic acinar to beta cells can be easily modulated by the administration of doxycycline. These mice will be used to explore how transdifferentiation actually occurs and to determine the extent to which the newly generated beta cells are functional and exhibit a gene expression profile similar to authentic beta cells. This project is based on the premise/hypothesis that multiple gene regulatory networks, which are normally set up during mouse development, must be established during directed or trans-differentiation of other cell types to achieve the beta cell-like functionalities necessary for clinical use. Greater knowledge of pancreas-specific gene regulatory networks, how they are established and differ among related cell populations, and determining whether they are present or absent in experimentally-derived cellular populations, will serve as a platform both for new discovery and protocol improvements. The six investigators in this project have a track record of productive collaborative interactions and bring specific knowledge and abilities necessary to accomplish these goals. Thus, it is anticipated that this project will generate important resources that will not only advance two of the overarching goals of the Beta Cell Biology Consortium but also have broad scientific impact and utility. PUBLIC HEALTH RELEVANCE: Type 1 and Type 2 diabetes are diseases that cause significant morbidity and mortality and thus have an adverse economic impact. Both diseases are characterized by the destruction or dysfunction of insulin-secreting pancreatic beta cells. This application seeks to gain key information for developing new, cell-based replacement therapies that hold promise for achieving better glucose control than is currently possible.