Oocyte factors for reprogramming to pluripotency

描述(由申请人提供)：本申请涉及干细胞的广泛挑战领域(14)和具体的挑战主题“14-HD-102：识别卵母细胞的重编程因素。”在这项提案中，我们基于我们在卵母细胞到胚胎转化、干细胞和重编程方面的现有数据、信息学和实验专业知识，基于我们关于人和小鼠卵母细胞到胚胎转化的数据，启动了一个人类细胞重编程的创新计划。我们有一套基于单卵母细胞转录图谱和其他正在筹备中的基因准备测试重新编程，生物信息学基础设施已经到位，但需要通过招聘新员工来恢复活力，以及有效的研究计划。我们的假设是，一组选择性转录本的变化导致了小鼠和女性卵母细胞的细胞质成熟，使它们转变为能够对配子和体细胞核进行重新编程的状态。为了解决这一假设，我们提出了三个特定的目标：1)用促黄体生成素(L H)和卵巢旁分泌激素处理能够刺激细胞质成熟的单个小鼠和人卵母细胞前后的基因表达。2)监测卵母细胞发育到囊胚期，通过基因操作抑制或增加我们认为在细胞质成熟过程中上调的关键卵母细胞基因的表达。3)用“传统因子”加/减我们的候选增强剂对我们的更新生物库中的体细胞进行重新编程。我们注意到，所有目标都在进行中，并可在两年的时间框架内交付。由斯坦福医学院、斯坦福医院和诊所以及露西尔·帕卡德儿童医院组成的斯坦福医学院的工作和发现，不仅使当前和未来的患者广泛受益，也使当地经济受益。2006年，根据一项独立顾问的研究，该大学向我们当地的两个县--加利福尼亚州的圣克拉拉县和圣马特奥县--提供了超过21亿美元的资金，仅这两家医院就产生了4.44亿美元的经济影响。斯坦福医学的发现导致了硅谷数十家公司的成立，即使在当前的经济危机中，这些公司仍在继续提供就业机会，刺激经济。我们目前的建议旨在确定细胞质成熟卵母细胞中的重新编程因素，以我们丰富的专业知识和初步数据为基础，从而使我们能够在两年的时间框架内迅速采取行动，推动科学和技术向前发展，并在就业、知识和推动私人技术投资的潜力方面为我们的经济做出贡献。我们估计，这项提议将对经济产生积极影响，为斯坦福医学创造4个工作岗位，帮助保留另外4个工作岗位，并间接创造额外的10到20个工作岗位(根据加州生物医学行业的数据，生物医学组织的每一名员工，将有另外3到5个工作岗位被服务于该行业的公司雇用)。 与公共卫生相关：该项目有三个目标，旨在探索这样一种假设，即一组选择性转录本的变化导致小鼠和女性卵母细胞的细胞质成熟，将它们转变为能够对配子细胞核和体细胞核进行重新编程的状态。该项目需要识别更多的重编程基因，验证卵母细胞的功能，并测试重编程的增强。我们建立在大量初步数据集的基础上，使我们能够通过为拟议的研究提供更多资金和人员，立即同时实现所有目标。

DESCRIPTION (provided by applicant): This application addresses the broad Challenge Area of Stem Cells (14) and specific Challenge Topic "14-HD- 102: Identifying Reprogramming Factors for Oocytes." In this proposal we build upon our current data, informatics and experimental expertise in the oocyte to embryo transition, stem cells and reprogramming, to initiate an innovative program in reprogramming of human cells based upon our data on the human and mouse oocyte to embryo transition. We have a set of genes ready to test for reprogramming based on single oocyte transcriptional profiling and others in the pipeline, a bioinformatics infrastructure in place but needing rejuvenation through new hires, and an effective research plan. Our hypothesis is that changes in a selective set of transcripts are responsible for the cytoplasmic maturation of oocytes in mice and women, transforming them to a state capable of reprogramming both gametic and somatic cell nuclei. To address this hypothesis, we propose three specific aims to: 1) Profile gene expression in individual mouse and human oocytes before and after treatment with leutinizing hormone (LH) and ovarian paracrine hormones capable of stimulating cytoplasmic maturation. 2) Monitor oocyte development to blastocyst stage following genetic manipulations to suppress or increase expression of key oocyte genes that we identify as upregulated during cytoplasmic maturation. 3) Reprogram somatic cells from our RENEW BioBank with "traditional factors" plus/minus our candidate enhancers. We note that all aims are underway and deliverable in a two-year time frame. The work and discoveries made at Stanford Medicine, composed of the Stanford School of Medicine, Stanford Hospital and Clinics and the Lucile Packard Children's Hospital, widely benefit not only current and future patients but also the local economy. In 2006, according to an independent consultant's study, the university provided more than $2.1 billion into our two local counties, Santa Clara and San Mateo counties in California, and the economic impact of the two hospitals alone was $444 million. Stanford Medicine discoveries have led to the creation of dozens of companies in Silicon Valley, which continue providing jobs and stimulating the economy even in the current economic crisis. Our current proposal, aimed at identifying reprogramming factors in cytoplasmically-mature oocytes builds upon our substantial expertise and preliminary data, thus allowing us to move quickly within the two-year time frame to move the science and technology forward and contribute to our economy in terms of jobs, knowledge and potential to fuel private technological investment. We estimate that this proposal would positively impact the economy by creating 4 jobs at Stanford Medicine, contributing to the retention of 4 others, and contributing to the creation of an additional 10 to 20 indirectly (according to the California Biomedical Industry, for every one employee of a biomedical organization, another three to five will be employed in firms that service that industry). PUBLIC HEALTH RELEVANCE: This project has three aims to explore the hypothesis that changes in a selective set of transcripts are responsible for the cytoplasmic maturation of oocytes in mice and women, transforming them to a state capable of reprogramming both gametic and somatic cell nuclei. The project entails identification of additional reprogramming genes, verifying oocyte function, and testing enhancement of reprogramming. We build on a substantial preliminary data set that allows us to immediately progress on all aims simultaneously through further funding and staffing of the proposed research.