Oocyte factors for reprogramming to pluripotency

重编程为多能性的卵母细胞因子

• 批准号: 7815481
• 负责人: AARON JW HSUEH
• 金额: $ 99.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-19 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7815481
• 关键词: Address; Aftercare; Area; Beginning of Human Life; Bioinformatics; California; Cell Nucleus; Cell fusion; Cells; Clinics and Hospitals; Consent; County; Data; Data Set; Development; Economics; Embryo; Embryonic Development; Employee; Enhancers; Epigenetic Process; Funding; Future; Gene Activation; Gene Expression; Gene Silencing; Genes; Hormones; Hospitals; Human; Human Development; Individual; Industry; Informatics; Injection of therapeutic agent; Investments; Knowledge; Life; Luteinizing Hormone; Medicine; Methylation; Microfluidics; Modification; Monitor; Mus; Occupations; Oocytes; Organism; Ovarian; Ovarian hormone; Pathway interactions; Patients; Pediatric Hospitals; Pharmacologic Substance; Preclinical Drug Evaluation; Primates; Process; RNA Degradation; Rana; Rejuvenation; Research; Research Infrastructure; Safety; Sampling; Science; Services; Silicon; Somatic Cell; Staging; Stem cells; System; Technology; Testing; Therapeutic; Time; Transcript; Transcriptional Activation; Universities; Vesicle; Woman; Work; base; biobank; blastocyst; cell bank; clinical application; demethylation; economic impact; egg; experience; gene function; genetic manipulation; imprint; innovation; medical schools; novel; oocyte maturation; overexpression; paracrine; pluripotency; programs; public health relevance; response; single cell analysis; somatic cell nuclear transfer; sperm cell; stem cell biology; vector; zygote

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.

描述（由申请人提供）：本申请涉及干细胞（14）的广泛挑战领域和特定挑战主题“14- hd - 102：识别卵母细胞的重编程因子”。在本提案中，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，基于我们在人类和小鼠卵母细胞到胚胎转化方面的数据，启动一个创新的人类细胞重编程项目。我们已经准备好了一组基因，可以基于单个卵母细胞转录谱和其他正在进行的基因重编程测试，生物信息学基础设施已经到位，但需要通过新员工来振兴，以及一个有效的研究计划。我们的假设是，一组选择性转录本的变化负责小鼠和女性卵母细胞的细胞质成熟，将它们转化为能够重新编程配子细胞核和体细胞细胞核的状态。为了解决这一假设，我们提出了三个具体目标：1)在使用亮氨酸生成素（LH）和卵巢旁分泌激素刺激细胞质成熟之前和之后，分析个体小鼠和人类卵母细胞的基因表达。2)通过基因操作监测卵母细胞发育到囊胚期，抑制或增加在细胞质成熟过程中被上调的关键卵母细胞基因的表达。3)用“传统因子”加上/减去我们的候选增强因子，对我们RENEW生物银行的体细胞进行重编程。我们注意到，所有目标都在进行中，并可在两年时间内实现。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所做的工作和发现，不仅广泛造福于现在和未来的病人，也造福于当地经济。根据一份独立顾问的研究，2006年，该校向加州的圣克拉拉县和圣马特奥县这两个地方县提供了超过21亿美元的资金，仅这两家医院的经济影响就达到4.44亿美元。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并刺激经济。我们目前的建议，旨在确定细胞质成熟卵母细胞中的重编程因子，建立在我们丰富的专业知识和初步数据的基础上，从而使我们能够在两年的时间框架内迅速行动，推动科学技术向前发展，并在就业、知识和潜力方面为我们的经济做出贡献，以推动私人技术投资。我们估计，这一提议将对经济产生积极影响，在斯坦福医学院创造4个就业机会，有助于留住另外4个就业机会，并间接创造10到20个就业机会（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，又会有3到5名员工受雇于为该行业服务的公司）。