Investigating regulators controlling differentiation potential of ES cells

研究控制 ES 细胞分化潜能的调节因子

• 批准号: 10237975
• 负责人: Jonghwan Kim
• 金额: $ 32.49万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10237975
• 关键词: Address; Adult; Affect; Apoptosis; Award; Biomedical Research; CASP3 gene; Caspase; Cell Death; Cell Density; Cell Fate Control; Cell Survival; Cell Therapy; Cells; Cessation of life; Characteristics; Decision Making; Disease model; ES Cell Line; Embryo; Embryonic Development; Enhancers; Environment; Epigenetic Process; Factor Analysis; Foundations; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genetic Transcription; Genomics; Heterogeneity; In Vitro; Individual; Knowledge; Light; Link; Mechanics; Mediating; Metabolic; MicroRNAs; Molecular; Monitor; National Institute of General Medical Sciences; Organism; Outcome; Pathway interactions; Pluripotent Stem Cells; Population; Proliferating; Proteins; Regulation; Reporter; Reproducibility; Research; Research Support; Role; Signal Pathway; Signal Transduction; Stem Cell Development; Techniques; Testing; Therapeutic; Tissue-Specific Gene Expression; Work; YY1 Transcription Factor; beta catenin; cell behavior; cell fate specification; cell type; density; drug discovery; embryonic stem cell; induced pluripotent stem cell; insight; instrument; interest; neglect; novel; overexpression; pluripotency; programs; protein expression; self-renewal; single-cell RNA sequencing; stem; stem cell differentiation; stem cell fate; stem cell self renewal; stem cell therapy; stem cells; tool; transcription factor; transcriptome

SUMMARY Pluripotent stem cells, such as embryonic stem (ES) cells and induced pluripotent stem (iPS) cells can proliferate indefinitely in vitro without changes in their characteristics (self-renewal) while keeping their potential to give rise to almost all cell types in adult organisms (pluripotency). Due to such exceptional characteristics, ES and iPS cells have been extensively studied and used as tools for understanding the molecular basis of early embryo development and also serve as useful instruments in drug discovery and establishing various disease models. To fully utilize their potential in therapeutic applications, it is crucial to completely understand how these two unique characteristics are modulated. Prior studies have largely focused on understanding of self-renewal, allowing us to better illuminate the regulatory mechanisms mediated by key transcription factors (TFs), signaling pathways, and other associated genomic features. On the other hand, understanding of exit mechanisms from self-renewal towards cell fate specification, and factors involved in proper differentiation of pluripotent stem cells have not yet been systematically examined. The long-term objective of the proposed research is to investigate regulators controlling differentiation potential of pluripotent stem cells. In our previous research supported by NIGMS awards, we have revealed multiple TFs, epigenetic regulators, and genomic features that influence the differentiation potential of ES cells. Among those factors, we showed that Yap1, a transcriptional co- regulator, downstream of the Hippo pathway, is dispensable for self-renewal but required for differentiation of ES cells. We furthermore revealed the roles of Yap1 in safeguarding ES cells from excessive cell death during differentiation. We additionally observed that cell density, tightly linked to the Hippo signaling activity, significantly affects global gene expression programs of not only self-renewing ES cells, but also their differentiation potential. However, underlying mechanisms of ES cell differentiation in the context of cell density and survival vs. death decision have been elusive. To address this critical gap in knowledge, our objectives of the proposal will be 1) to determine, at the single cell level, how the survival vs. death decision is made when ES cells differentiate, 2) to define outcomes of density- dependent gene expression signatures and enhancer usage during ES cell self-renewal and differentiation, and 3) to identify effectors controlling density-dependent gene expression programs and elucidate their regulatory mechanisms. The information obtained from the proposal will provide novel insights into the reproducibility issues in biomedical studies caused by inconsistencies in cell density between different experimental techniques. Furthermore, outcomes of this proposal will provide a foundation for manipulation of stem cells to control cell fates towards desired lineages and contribute to the advances in stem cell-based cell therapies.

概括 多能干细胞，例如胚胎干（ES）细胞和诱导多能干（iPS）细胞可以 在体外无限增殖而不改变其特性（自我更新），同时保持其特性 具有在成体生物体中产生几乎所有细胞类型的潜力（多能性）。由于如此特殊的 ES 和 iPS 细胞的特征、ES 和 iPS 细胞已被广泛研究并用作了解 早期胚胎发育的分子基础，也可作为药物发现和药物发现的有用工具 建立各种疾病模型。为了充分发挥其在治疗应用中的潜力，至关重要 完全理解这两个独特的特性是如何调制的。先前的研究很大程度上 聚焦于对自我更新的理解，让我们更好地阐明调控机制 由关键转录因子 (TF)、信号​​通路和其他相关基因组特征介导。 另一方面，了解从自我更新到细胞命运规范的退出机制， 参与多能干细胞正确分化的因素尚未得到系统的研究 检查了。拟议研究的长期目标是调查监管机构控制 多能干细胞的分化潜能。在我们之前获得 NIGMS 奖项支持的研究中， 我们已经揭示了多种影响转录因子、表观遗传调节因子和基因组特征 ES细胞的分化潜能。在这些因素中，我们发现 Yap1，一种转录共 Hippo 通路下游的调节器对于自我更新来说是可有可无的，但对于自我更新来说是必需的 ES细胞的分化。我们还揭示了 Yap1 在保护 ES 细胞免受 分化过程中细胞过度死亡。我们还观察到细胞密度与 Hippo 信号传导活动，不仅显着影响自我更新的全局基因表达程序 ES细胞，还有它们的分化潜力。然而，ES细胞的潜在机制 细胞密度和生存与死亡决策方面的差异一直难以捉摸。致地址 对于这一知识上的关键差距，我们提案的目标将是 1) 在单细胞水平上确定， ES 细胞分化时如何做出生存与死亡决策，2) 定义密度结果- ES细胞自我更新过程中依赖的基因表达特征和增强子的使用 分化，3) 识别控制密度依赖性基因表达程序的效应子和 阐明其调控机制。从提案中获得的信息将提供新颖的 深入了解生物医学研究中由细胞密度不一致引起的重现性问题 不同的实验技术之间。此外，该提案的结果将提供 为操纵干细胞以控制细胞命运向所需谱系奠定基础，并有助于 基于干细胞的细胞疗法的进展。