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）：此申请涉及干细胞（14）和特定挑战主题的广泛挑战领域“ 14-HD-102：确定卵母细胞的重编程因素”。在这项建议中，我们基于卵母细胞的当前数据，信息学和实验专业知识，以胚胎过渡，干细胞和重编程，以根据我们对人类和小鼠卵母细胞的数据重编程人类细胞进行重编程的创新程序，以启动胚胎过渡。我们准备了一组基因，可以根据单个卵母细胞转录分析进行重新编程，以及其他中的其他基因，生物信息学基础架构的到位，但需要通过新员工重新焕发活力以及有效的研究计划。我们的假设是，选择性转录本中的变化是导致小鼠和女性卵母细胞的细胞质成熟，从而将其转化为能够重编程配子和体细胞核的状态。为了解决这一假设，我们提出了三个特定的目的：1）在用甲状腺激素（LH）和卵巢旁分泌激素治疗之前和之后，单个小鼠和人类卵母细胞中的谱图基因表达，能够刺激细胞质成熟。 2）在遗传操作后，监测卵母细胞的发育至胚泡阶段，以抑制或增加我们在细胞质成熟过程中鉴定为上调的关键卵母细胞基因的表达。 3）从我们的续订生物库中重新编程具有“传统因素”加/减去候选增强子的体细胞。我们注意到，所有目标都在进行两年的时间范围内。斯坦福大学医学院，斯坦福医院和诊所以及卢西尔·帕卡德儿童医院的工作和发现，不仅受益于当前和未来的患者，而且在当地经济中受益。根据一项独立顾问的研究，2006年，该大学向我们的两个当地县分别提供了超过21亿美元的加利福尼亚州圣克拉拉和圣马特奥县，仅这两家医院的经济影响是4.44亿美元。斯坦福医学的发现导致在硅谷成立了数十家公司，这些公司仍在继续提供就业机会并刺激经济，即使在当前的经济危机中也是如此。我们目前的提案旨在确定细胞质成熟卵母细胞中的重编程因素建立在我们的大量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范围内迅速发展，从而推动科学和技术向前发展，并在工作，知识和潜力促进私人技术投资方面为我们的经济做出了贡献。我们估计，这项提案将通过在斯坦福医学上创造4个就业机会，为保留4个工作做出贡献，并为另外10至20的创造做出贡献（根据加利福尼亚生物医学行业的增加，为生物医学组织的每个雇员说，另外三到五个在该行业的公司中都将雇用3至5个）。 公共卫生相关性：该项目的三个旨在探讨一个假设，即选择性成绩单中的变化是导致小鼠和女性卵母细胞的细胞质成熟，从而将其转化为能够重新编程Gamentic和Sompation Cell Nuclei的状态。该项目需要识别其他重编程基因，验证卵母细胞功能以及测试重编程的增强。我们建立在一个实质性的初步数据集的基础上，该数据集使我们能够通过进一步的资金和拟议研究人员的身份立即在所有目标上进步。