Utilizing single cell approaches to decipher the mechanism underlying somatic cel

描述(由申请人提供)：了解正在重新编程为多能性的体细胞的基本分子元素是推动干细胞研究走向临床应用的先决条件。该领域的一个重要问题是，重新编程过程是由不按特定顺序发生的随机事件驱动的，还是更具确定性的过程，具有一定的转录变化模式，必须按顺序发生才能将分化的细胞转化为诱导多能干细胞(IPSCs)。破译这些元素的主要障碍之一是转导细胞群体的异质性，以及最终将产生IPSCs的一小部分细胞。该领域的大多数研究都是基于细胞群体分析的，这一事实有力地表明，利用单细胞技术对于理解重新编程过程至关重要。因此，我博士后培训的主要目标将是使用一种监测单细胞转录图谱的创新单细胞技术来破译定义重新编程过程的分子机制。这项技术将让我了解在重新编程过程中发生的主要事件是什么，它们发生的时间顺序是什么，以及哪些基因负责执行这些事件。然后，我将描述将在我的分析中突出显示的事件和关键参与者的特征，以了解它们如何在重新编程过程中执行其功能。最后，我将使用突出显示的基因生成敲入二级系统，用于未来的重新编程研究。解决这些问题将有助于阐明重新编程过程背后的分子机制，并将成为确定重新编程过程是确定性还是随机性的有力工具。 公共卫生相关性：终末分化细胞可以改变它们的命运成为ES样细胞(即重新编程过程)，这一发现为药物筛选、医学研究和患者特定细胞治疗提供了宝贵的资源。这项拟议的研究将阐明定义“真正的”重新编程过程的机制。

DESCRIPTION (provided by applicant): Understanding the basic molecular elements that characterize somatic cells undergoing reprogramming to pluripotency is a prerequisite for advancing stem cell research towards clinical applications. One of the important questions in the field is whether the reprogramming process is driven by stochastic events that occur in no particular sequence or rather it is a more deterministic process that has a defined pattern of transcriptional changes that must occur sequentially to convert the differentiated cells to induced pluripotent stem cells (iPSCs). One of the major obstacles in deciphering these elements is the heterogeneity of the transduced cell population and the small fraction of cells that eventually will yield iPSCs. The fact that the majority of the studies in the field are based on cell-population analyses, which presumably do not represent the small fraction of cells that undergo reprogramming, strongly suggests that utilizing single cell techniques will be of paramount importance to understand the reprogramming process. Therefore, the main objective of my postdoctoral training will be to decipher the molecular mechanisms that define the reprogramming process using an innovative single cell technique that monitors the transcriptional profile of single cells. This technique will allow me to understand what the main events that occur during the reprogramming process are, in what chronological order they occur, and which genes are responsible for executing these events. I will then characterize the events and the key players that will be highlighted in my analysis to understand how they execute their function in the reprogramming process. Finally, I will generate knock-in secondary systems using the highlighted genes for future reprogramming studies. Addressing these questions will shed light on the molecular mechanisms that underlie the reprogramming process and will be a powerful tool to determine whether the reprogramming process is deterministic or stochastic. PUBLIC HEALTH RELEVANCE: The discovery that terminally differentiated cells can change their fate to become ES-like cells (i.e. reprogramming process) provides an invaluable resource for drug screening, medical research, and patient specific cell-based therapy. The proposed research will shed light on the mechanisms that define the "true" reprogramming process.