Transcriptional Regulatory Networks in Living Cells

描述(申请人提供)：胚胎干细胞(ES)和诱导多能干细胞(IPS)具有巨大的医疗应用潜力，主要受限于我们将这些细胞的活动引导到特定治疗目标的方法。更全面地了解多能干细胞的调节电路几乎肯定会提供有助于克服这一限制的见解。我们建议通过鉴定新的ES细胞转录因子、染色质调节因子和信号蛋白，并确定它们如何共同作用来控制负责多能性和自我更新的基因表达程序，从而扩大对小鼠和人类胚胎干细胞转录调控电路的了解。此外，我们建议利用对ES细胞状态控制的更好的理解来开发更强大的细胞重编程方法来产生iPS细胞。为了实现这些目标，该建议的具体目标是：1)结合并进一步开发强大的实验和分析技术，以确定ES细胞状态的新型调节因子并确定其在全基因组的占有率和功能；2)确定在小鼠和人类ES细胞的识别中发挥关键作用的新型转录因子、染色质调节因子和信号蛋白；3)确定新型转录因子、染色质调节因子和信号蛋白如何促进小鼠和人类ES细胞的多能性和自我更新；以及4)利用细胞重编程试验来深入了解细胞状态的控制并开发更有效的细胞重编程方法。从这些研究中对转录调控电路的了解将带来对ES细胞状态控制的新见解，揭示关键调控因子如何控制ES细胞的基因表达程序，促进为再生医学操纵细胞命运的努力，并为进一步绘制人类和其他脊椎动物细胞的调控电路提供基础。 与公共健康相关：我们计划通过识别新的ES细胞调节器并确定它们如何发挥作用来控制这些多能细胞特有的基因表达程序，来扩大对胚胎干细胞调控电路的知识。胚胎干细胞在不同的医学应用中具有巨大的潜力，这些研究对调控电路的理解应该会促进为再生医学操纵细胞命运的努力。

DESCRIPTION (provided by applicant): Embryonic stem (ES) and induced pluripotent stem (iPS) cells hold enormous potential for diverse medical applications, limited primarily by our methods to direct the activities of these cells toward specific therapeutic goals. A more complete understanding of the regulatory circuitry of pluripotent stem cells would almost certainly provide insights useful to overcome this limitation. We propose to expand knowledge of the transcriptional regulatory circuitry of murine and human embryonic stem (ES) cells by identifying novel ES cell transcription factors, chromatin regulators and signaling proteins and by determining how they function together to control the gene expression program responsible for pluripotency and self-renewal. Furthermore, we propose to use improved understanding of the control of ES cell state to develop more powerful methods for cellular reprogramming to generate iPS cells. To accomplish these goals, the specific aims of the proposal are: 1) Combine and further develop powerful experimental and analytical technologies that can identify novel regulators of ES cell state and determine their genome-wide occupancy and function; 2) Identify novel transcription factors, chromatin regulators and signaling proteins that play key roles in murine and human ES cell identity; 3) Determine how novel transcription factors, chromatin regulators and signaling proteins contribute to pluripotency and self-renewal in murine and human ES cells; and 4) Use cellular reprogramming assays to gain insights into the control of cell state and to develop more powerful methods for cellular reprogramming. Improved understanding of transcriptional regulatory circuitry from these studies will lead to new insights into the control of ES cell state, reveal how key regulators control the gene expression program of ES cells, facilitate efforts to manipulate cell fates for regenerative medicine, and provide the foundation for further mapping regulatory circuitry in human and other vertebrate cells. PUBLIC HEALTH RELEVANCE: We plan to expand knowledge of the regulatory circuitry of embryonic stem cells by identifying novel ES cell regulators and determining how they function to control the gene expression program that is unique to these pluripotent cells. Embryonic stem cells hold enormous potential for diverse medical applications and improved understanding of regulatory circuitry from these studies should facilitate efforts to manipulate cell fates for regenerative medicine.