Microenvironmental cues control pancreas cell fate and beta-cell maturation

摘要 目前通过细胞替代治疗1型糖尿病的策略受到以下限制 供体胰岛短缺；人多能干细胞来源的β(hPSC-β)细胞 代表着解决这一困境的一个有希望的解决方案。要改进当前的方法，以 人骨髓间充质干细胞向功能性β细胞分化的研究 内分泌发育和β细胞发育过程的分子程序 成熟仍然是需要的。我们的初步研究表明，整合素信号和 微环境僵硬在协调胰岛和β细胞中的作用 发展。本提案试图阐明整合素和 微环境僵硬在协调这些复杂的过程中起着作用。我们的目标是 揭示整合素信号控制内分泌细胞命运的机制 决定，2)胰岛发育，3)β细胞成熟。我们将结合使用 胰腺、胰岛和β细胞特异性缺失小鼠模型灭活整合素亚单位 β1(Itgb1，整合素信号转导所必需的亚基)。我们的目标也是在体外促进 通过控制微环境硬度使hPSC-β细胞成熟。要实现 为此，我们建立了一种新型的细胞培养平台，用于时间控制 微环境刚性。通过培养和分化hPSC-β祖细胞 在这个平台上，我们将阐述微环境刚性对β-CELL的作用 成熟。总之，我们的研究将使我们更好地理解 整合素信号和微环境刚性在调节β细胞发育中的作用。这 将为未来功能性hPSC-β的生成策略提供信息。

ABSTRACT Current strategies for treating type 1 diabetes via cell replacement are limited by a shortage of donor islets; human pluripotent stem cell (hPSC)-derived β (hPSC-β) cells represent a promising solution to this dilemma. To improve current methodologies for the differentiation of hPSCs towards functional β-cells, a comprehensive understanding of the molecular programs for the processes of endocrine development and β-cells maturation is still required. Our preliminary studies suggested that integrin signaling and microenvironment stiffness play an integral role in coordinating islet and β-cell development. The present proposal seeks to elucidate the role that integrin and microenvironment stiffness play in coordinating these complex processes. We aim to uncover the mechanisms of integrin signaling in controlling 1) endocrine cell fate decision, 2) islet development, and 3) β-cell maturation. We will utilize a combination of pancreas-, islet-, and β-cell-specific deletion mouse models to inactivate integrin subunit β1 (Itgb1, a subunit essential for integrin signaling). We also aim to promote in vitro maturation of hPSC-β cells by controlling the microenvironments stiffness. To achieve this, we established a novel cell culture platform for temporal control of microenvironment stiffness. By culturing and differentiating the hPSC-β progenitors on this platform, we will elucidate the role which microenvironment stiffness plays on β-cell maturation. Together, our study will provide a greater understanding of the role of integrin signaling and microenvironment stiffness in regulating β-cell development. This will inform future functional hPSC-β generation strategies.