Formation of endocrine pancreas progenitors

描述（申请人提供）：用于1型糖尿病的基于细胞的疗法的方法主要集中在可以驱动基因表达的细胞特异性模式的单个转录因子或转录因子组合的表达，并且大多数至少部分依赖于Pdx 1，一种在早期胰腺发育和成熟细胞中具有关键调节作用的同源结构域转录因子，以及Ngn 3，一种关键的前内分泌转录因子，其瞬时表达是内分泌祖细胞的特征。无论多么有希望，通过这些方法中的任何一种获得细胞表型的效率仍然很低，并且这些努力的保真度和稳定性尚未得到充分评估。虽然内胚层可重复且有效地实现，但是随后的转变，例如从胰腺祖细胞到内分泌胰腺祖细胞，以特别低的效率实现。我们最近确定，Pdx 1有助于规范的内分泌祖细胞在小鼠中直接调节Ngn 3与Hnf 6和参与在早期胰腺发育过程中的交叉调节转录因子网络。我们假设，理解转录因子Ngn 3的表观遗传调控以及Pdx 1和Hnf 6在发育细胞的交叉调控转录因子网络中的作用，将有助于提高通过引导非细胞群体分化实现的细胞表型的效率、保真度和稳定性。因此，我们在AIM 1中提出：检查Ngn 3在人胰腺祖细胞中的遗传和表观遗传调节，在AIM 2中：分析Pdx 1和Hnf 6在促进内分泌祖细胞特化中的遗传和分子相互作用，在AIM 3中：通过全局占有和基因表达分析来确定Pdx 1和Hnf 6在内分泌胰腺祖细胞转录网络中的作用。首要的长期目标是利用从这些研究中获得的信息来优化ES和iPS细胞以及可能的成熟谱系向细胞命运的引导转变。 公共卫生相关性：对1型糖尿病细胞治疗的高度希望已经被有限的可获得性和短期功能的人类胰岛移植所冲淡，导致对替代来源的强烈关注。细胞所提出的研究的总体目标是优化ES和iPS细胞向细胞命运的引导转变，以使潜在的患者特异性细胞可用于移植。

DESCRIPTION (provided by applicant): Approaches to cell based therapies for type 1 diabetes have largely focused on the expression of a single or a combination of transcription factors that can drive ( cell specific patterns of gene expression, and most have relied at least in part on Pdx1, a homeodomain transcription factor with critical regulatory roles in early pancreas development and in the mature ( cell, and on Ngn3, a critical pro-endocrine transcription factor whose transient expression characterizes endocrine progenitors. However promising, the efficiency of achieving a ( cell phenotype by any of these approaches remains low, and the fidelity and stability of these efforts have not been fully evaluated. Although endoderm is achieved reproducibly and efficiently, subsequent transitions, such as from pancreatic progenitor to endocrine pancreatic progenitor, are achieved with particularly low efficiency. We recently determined that Pdx1 contributes to specification of endocrine progenitors in mice both by regulating Ngn3 directly in concert with Hnf6 and by participating in a cross-regulatory transcription factor network during early pancreas development. We hypothesize that understanding the epigenetic regulation of the transcription factor Ngn3 and the roles of Pdx1 and Hnf6 in the cross-regulatory transcription factor network of developing ( cells will inform efforts to improve the efficiency, fidelity and stability of the ( cell phenotype achieved through the guided differentiation of non-( cell populations. Therefore, we propose in AIM 1: To examine the genetic and epigenetic regulation of Ngn3 in human pancreas progenitors, in AIM 2: To analyze the genetic and molecular interactions between Pdx1 and Hnf6 in promoting endocrine progenitor specification, and in AIM 3: To define the role of Pdx1 and Hnf6 in the transcriptional network of endocrine pancreas progenitors through global occupancy and gene expression analysis. The overarching long-term goal is to utilize this information gained from these studies to optimize the guided transition of ES and iPS cells, and possibly mature lineages, toward a ( cell fate. PUBLIC HEALTH RELEVANCE: The high hopes for cell based therapy for type 1 diabetes have been tempered by the limited availability and short-term function of human islets for transplantation, leading to intense focus on alternate sources of (?cells. The overarching goal of the proposed studies is to optimize the guided transitions of ES and iPS cells toward a ( cell fate to make potentially patient-specific ( cells available for transplantation.