Oocyte factors for reprogramming to pluripotency

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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.