Genomic stability of human pluripotent stem cells

人类多能干细胞的基因组稳定性

• 批准号: 8447348
• 负责人: Paul Hubert Lerou
• 金额: $ 23.28万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8447348
• 关键词: Accounting; Affect; Aneuploidy; Award; Biological; Biological Assay; CDK2 gene; Cell Culture Techniques; Cell Cycle; Cell Cycle Regulation; Cell Cycle Regulation Pathway; Cell Line; Cell division; Cell model; Cells; Cellular biology; Chromosomes; Complement; Coupled; Cyclin E; Cytogenetic Analysis; DNA; DNA Damage; DNA Repair; DNA biosynthesis; Defect; Development; Disease; Embryo; Ensure; Environment; Fibroblasts; Funding; Future; Generations; Genes; Genetic; Genome Stability; Genomic Instability; Germ Cells; Goals; Hematology; Hospital Departments; Human; Imaging Techniques; Imaging technology; Infertility; Institutes; Institution; Instruction; Life; Maintenance; Malignant Neoplasms; Mediating; Mentors; Mentorship; Mitotic; Molecular; Mutation; Pathway interactions; Patients; Pediatric Hospitals; Phase; Pluripotent Stem Cells; Production; RNA Interference; Regulation; Reporter; Research; Research Proposals; Resources; Role; Scientist; Somatic Cell; Stem cells; Syndrome; Systems Biology; Techniques; Telomere Maintenance; Testing; Training; Training Programs; Viral; base; cancer genetics; career; cell fixing; cellular imaging; daughter cell; embryonic stem cell; human embryonic stem cell; improved; induced pluripotent stem cell; insight; knock-down; medical schools; novel; oncology; repaired; response; segregation; self-renewal; senescence; small molecule; stem; stem cell biology; telomere; tool; tumorigenesis

The cell cycle ensures faithful duplication and segregation of duplicated chromosomes into daughter cells. Dysregulation of the cell cycle results in genomic instability, including aneuploidy, a hallmark of cancer and a significant cause of infertility and genetic syndromes when aneuploidy arises in the early embryo or gametes. ES cells maintain genomic stability even though they have altered Rb and p53 function and they are able to escape senescence, dividing indefinitely without the accruing DNA damage. This research proposal will test the hypothesis that hES cells possess unique and specialized mechanisms to maintain genomic stability. My career goal is to become an independently funded clinician-scientist in the field of stem cell biology with expertise in hES and induced pluripotent stem (iPS) cell manipulation and cell cycle regulation. The pathway to Independence Award (K99/R00) will greatly assist my transition to full independence. In the immediate future, this award will allow me to gain in-depth expertise in pluripotent cell biology, live-cell imaging and the advanced molecular biological techniques to study cell cycle regulation under the mentorship of Dr. George Q. Daley and co-mentorship of Dr. Marc W. Kirschner. The training program in the hematology/oncology division at Children's Hospital and department of systems biology at Harvard Medical School brings together resources from both institutions as well as those ofthe Harvard Stem Cell Institute, and provides an outstanding environment for the completion of training during the mentored phase. This will greatly facilitate my effective transition to independence. ^ The specific aims of this research proposal are to: 1) characterize the Rb and p53 pathways and their role in genomic stability maintenance in human pluripotent cells, and 2) generate IPS cells from somatic cells that harbor mutations implicated in genomic instability. Advanced techniques, including live and fixed cell imaging, will be applied to human pluripotent stem cells to perform in depth analysis ofthe pathways maintaining genomic stability. Viral-mediated gene over expression / RNAi gene knock-down techniques and small molecules will be used to interrogate these pathways and will be complemented by the development genomic instability disease specific iPS cell lines. The goal of this research is to study the mechanisms of genomic stability maintenance in pluripotent cells in order to better understand the molecular basis of aneuploidy.

细胞周期确保复制的染色体忠实地复制和分离到子细胞中。 细胞周期失调会导致基因组不稳定，包括非整倍性，这是癌症和癌症的一个标志。 当早期胚胎或配子中出现非整倍性时，这是不育和遗传综合征的重要原因。 ES 细胞即使改变了 Rb 和 p53 功能，仍保持基因组稳定性，并且能够 逃避衰老，无限期分裂，而不会造成 DNA 损伤。本研究计划将测试 假设 hES 细胞拥有独特且专门的机制来维持基因组稳定性。 我的职业目标是成为干细胞生物学领域的一名独立资助的临床医生科学家 在 hES 和诱导多能干 (iPS) 细胞操作和细胞周期调控方面拥有丰富的专业知识。 获得独立奖（K99/R00）的途径将极大地帮助我过渡到完全独立。在 在不久的将来，这个奖项将使我获得多能细胞生物学、活细胞方面的深入专业知识 成像和先进的分子生物学技术来研究细胞周期调控 George Q. Daley 博士的指导和 Marc W. Kirschner 博士的共同指导。培训计划在 儿童医院血液学/肿瘤科和哈佛医学院系统生物学系 学校汇集了两个机构以及哈佛干细胞研究所的资源， 并为指导阶段完成培训提供良好的环境。这将 极大地促进了我向独立的有效过渡。 ^ 本研究计划的具体目标是：1) 表征 Rb 和 p53 通路及其在 维持人类多能细胞的基因组稳定性，2) 从体细胞产生 IPS 细胞 存在与基因组不稳定有关的突变。先进技术，包括活细胞和固定细胞 成像，将应用于人类多能干细胞，对通路进行深入分析 维持基因组稳定性。病毒介导的基因过表达/RNAi基因敲低技术和 小分子将被用来探究这些途径，并将通过开发来补充 基因组不稳定疾病特异性 iPS 细胞系。 本研究的目的是研究多能细胞基因组稳定性维持的机制。 为了更好地了解非整倍体的分子基础。