Transcription Factor Induced Reprogramming

转录因子诱导的重编程

• 批准号: 8042392
• 负责人: Konrad Hochedlinger
• 金额: $ 32.89万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-29 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8042392
• 关键词: Transcription; Factor; Induced; Reprogramming

DESCRIPTION (provided by applicant): This application seeks to investigate the mechanisms underlying cellular reprogramming, i.e. the conversion of adult cells into pluripotent stem cells. We will utilize transcription factor-mediated reprogramming into induced pluripotent stem cells (iPSCs) as a tool to pursue three major Aims. In Aim 1, we will (i) test whether iPSCs derived from fibroblasts, hematopoietic and myogenic cells are transcriptionally, epigenetically and functionally distinct, (ii) study the mechanism for the increased reprogramming efficiency of progenitors, and (iii) determine if iPSC formation introduces genetic mutations into cells. Addressing the epigenetic and genetic integrity of iPSCs will be crucial for any potential therapeutic applications of this technology and may identify the most suitable cell type for generating patient-specific iPSCs. In Aim 2, we will (i) map the transcriptional and epigenetic changes in intermediate cell populations undergoing reprogramming, (ii) test if reprogramming differentiated cells into pluripotent cells recapitulates stages of normal development, and (iii) perform a gain and loss-of-function screen, respectively, to identify novel regulators of reprogramming. This aim will identify new molecules important during reprogramming, whose manipulation may facilitate the efficient and safe generation of patient-specific iPSCs. In Aim 3, we will investigate the functionality of iPSCs compared with ESCs using in vitro and in vivo assays. Specifically, we will (i) assess whether neural stem cells and fibroblasts derived from iPSCs and ESCs show similar growth and differentiation characteristics in vitro, (ii) test if iPSC-derived hematopoietic stem cells are as potent as ESC-derived hematopoietic stem cells upon serial bone marrow transplantation, and (iii) produce and age entirely ESC and iPSC-derived mice to test if iPSC-derived animals age prematurely or develop cancer. This aim will assess the safety and long-term consequences of iPSCs-derived mature cells in vivo, a prerequisite for using iPSC technology in human cell therapy. PUBLIC HEALTH RELEVANCE: The goal of our lab is to dissect the mechanisms of cellular reprogramming by using induced pluripotent stem cells (iPSCs) as a tool. In this proposal, we will (i) study the role of the somatic cell-of-origin during cellular reprogramming into iPSCs, (ii) map the transcriptional and epigenetic events that occur in intermediate cells undergoing reprogramming, and (iii) compare the developmental and differentiation potentials of iPSCs and embryonic stem cells. Collectively, these experiments will give insight into the mechanisms of transcription-factor-mediated reprogramming and provide crucial information on the efficacy and safety of iPSC production for studying and potentially treating diseases.

描述（由申请人提供）：本应用程序旨在研究细胞重编程的基础机制，即成年细胞转化为多能干细胞。我们将使用转录因子介导的重编程为诱导的多能干细胞（IPSC）作为追求三个主要目标的工具。 在AIM 1中，我们将（i）测试源自成纤维细胞，造血和肌生成细胞的IPSC是否在转录，表观遗传和功能上是不同的，（ii）研究了祖细胞重编程效率提高的机制，以及（iii）确定IPSC是否将遗传突变引入细胞中。解决IPSC的表观遗传和遗传完整性对于该技术的任何潜在治疗应用至关重要，并可能确定最合适的细胞类型用于生成患者特异性IPSC。 在AIM 2中，我们将（i）绘制经过重编程的中间细胞群体的转录和表观遗传变化，（ii）测试如果将重编程分化为多能细胞中的分化细胞缩减了正常发育阶段，并且（iii）分别执行了功能障碍屏幕，以识别重编程的新型调节者。该目标将确定重新编程中重要的新分子，其操作可以促进患者特异性IPSC的有效生成。 在AIM 3中，我们将研究IPSC的功能与使用体外和体内测定的ESC相比。具体而言，我们将（i）评估来自IPSC和ESC的神经干细胞和成纤维细胞在体外表现出相似的生长和分化特征，（ii）测试是否具有IPSC衍生的造血干细胞是否有效，是否像ESC衍生的造血干细胞一样有效IPSC衍生的动物过早年龄或发展癌症。该目标将评估体内IPSC衍生的成熟细胞的安全性和长期后果，这是在人类细胞疗法中使用IPSC技术的前提。 公共卫生相关性：我们实验室的目的是通过使用诱导多能干细胞（IPSC）作为工具来剖析细胞重编程的机制。在此提案中，我们将（i）研究细胞重编程过程中的体细胞细胞在IPSC中的作用，（ii）绘制经过重编程的中间细胞中发生的转录和表观遗传事件，以及（iii）比较IPSC和胚胎干细胞的发育和分化势。总的来说，这些实验将深入了解转录因子介导的重编程的机制，并提供有关IPSC生产研究和潜在治疗疾病的功效和安全性的关键信息。