Epitenetic regulation and enhancement of beta cell differentiation

描述（由申请人提供）：人胚胎干细胞（hESCs）可能为1型糖尿病的细胞替代治疗提供胰岛素分泌β样细胞的无限来源，1型糖尿病是一种自身免疫破坏朗格汉斯胰岛分泌胰岛素的β细胞的疾病。尽管最近在从hESCs中获得早期终代和胰腺内胚层样祖细胞方面取得了进展，但尚未在体外获得强大的晚期功能胰腺β样细胞。人们越来越认识到，染色质结构的深刻改变，包括表观遗传DNA甲基化和组蛋白修饰的变化，有助于控制细胞命运的决定。为了利用hESCs在体外有效分化的能力，有必要进一步定义胰腺发育过程中的表观遗传特征。在这个β细胞生物学联盟的申请中，我们建议确定和开发新的研究资源和工具，以推进我们的长期目标，即提高从培养的hESCs中获得临床应用的人类胰腺β样细胞的产量，用于1型糖尿病的细胞替代治疗。特异性目标1是对两种细胞群体进行DNA甲基化和关键组蛋白标记的表观基因组分析：一种是来自尸体器官的成熟成人β细胞，另一种是来自人类esc的胰腺内分泌样祖细胞，其表达神经原素3（一种形成内分泌胰腺所需的转录因子）。然后，我们将比较这两个群体的表观遗传特征，这将导致表观遗传目标的识别，并允许开发策略来操纵细胞分化。特异性目的2是提高体外hESCs中功能β样细胞的产量。我们将使用三种新兴技术，即大分子水凝胶，蛋白质转导域和聚酰胺，来传递或抑制文献中已知的影响β细胞承诺，成熟和功能的特定分子。如果成功完成，这项工作将是第一个描述不同人类胰腺细胞群表观遗传谱的数据集。所采用的新技术也将对社区在β细胞生物学领域的进一步探索有价值。最后，这项工作将使我们能够以一种新的方式评估hesc来源的细胞，增加我们对这些细胞是否适合未来临床应用的理解。

DESCRIPTION (provided by applicant): Human embryonic stem cells (hESCs) may provide an unlimited source of insulin-secreting beta-like cells for cell replacement therapy of type 1 diabetes, a disease in which insulin-secreting beta cells in the pancreatic islets of Langerhans are destroyed by autoimmunity. In spite of the recent progress in deriving early definitive and pancreatic endoderm-like progenitors from hESCs, a robust generation of later functional pancreatic beta-like cells has not yet been achieved in vitro. It is increasingly recognized that profound alterations in chromatin structure, including changes in epigenetic DNA methylation and histone modifications, contribute to the control of cell fate decisions. To harness the capability to efficiently differentiate hESCs in vitro, it is necessary to further define epigenetic signatures during pancreatic development. In this Beta Cell Biology Consortium application, we propose to identify and develop novel research resources and tools to advance our long-term goal of enhancing the yield of clinically-applicable human pancreatic beta-like cells from hESCs in culture for the purpose of cell replacement therapy for type 1 diabetes. Specific Aim 1 is to perform epigenome profiling for DNA methylation and key histone marks on two cell populations: mature adult human beta cells, obtained from cadaverous organs, and human ESC-derived pancreatic endocrine like progenitor cells that express neurogenin 3, a transcription factor necessary for the formation of the endocrine pancreas. We will then compare the epigenetic signature of these two populations, which will lead to identification of epigenetic targets and allow development of strategies to manipulate cell differentiation. Specific Aim 2 is to improve the yield of functional beta like cells from hESCs in vitro. We will use three emerging technologies, i.e., macromolecular hydrogels, protein transduction domains, and polyamides, to deliver or inhibit specific molecules that are known, in the literature, to affect beta cell commitment, maturation and function. If successfully completed, the proposed work will be the first data set describing the epigenetic profiles of different human pancreatic cell populations. The novel technologies to be employed will also be valuable for the community to further explore in the field of beta cell biology. Finally, the proposed work will allow us to assess hESC-derived cells in a novel manner, increasing our understanding about whether these cells may be suitable for future clinical applications. PUBLIC HEALTH RELEVANCE: In spite of the recent progress in deriving early definitive and pancreatic endoderm-like progenitors from human embryonic stem cells (hESCs), a robust generation of later functional pancreatic beta-like cells has not yet been achieved in culture. We propose to identify and develop novel research resources (such as epigenetic signatures of beta cells and progenitors) and tools (such as macromolecular hydrogels, protein transduction domains, and polyamides) to advance our long-term goal of enhancing the yield of clinically-applicable human pancreatic beta-like cells from hESCs in culture for the purpose of cell replacement therapy for type 1 diabetes.