Transcriptional regulatory networks in spermatogonial stem cells

精原干细胞的转录调控网络

• 批准号: 8323107
• 负责人: Brian Peter Hermann
• 金额: $ 23.69万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8323107
• 关键词: Address; Adult; Area; Binding; Bioinformatics; Biological; Biological Assay; Cell Culture Techniques; Cells; Commit; Complex; Cytokinesis; Data; Data Analyses; Development; Development Plans; Doctor of Philosophy; EMSA; Environment; Faculty; Fertility; Foundations; Funding; Future; Gene Combinations; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Transcription; Germ Cells; Goals; Infertility; Instruction; K-Series Research Career Programs; Letters; Life; Luciferases; Male Infertility; Mentors; Mentorship; Methods; Mitosis; Molecular; Mus; Pathway interactions; Pattern; Phase; Plants; Positioning Attribute; Primates; Principal Investigator; Process; Production; Productivity; Program Evaluation; Property; Proteins; Regulation; Regulator Genes; Regulatory Element; Reporter; Reproductive Biology; Reproductive Physiology; Research; Research Institute; Research Personnel; Resources; Rodent; Role; Running; Science; Secure; Site; Spermatogenesis; Spermatogonia; Stem cell transplant; Stem cells; Structure; Subfamily lentivirinae; T-cell acute lymphocytic leukemia 1 protein; Testing; Testis; Tissues; Training; Transcriptional Regulation; Transfection; Translations; Transplantation; Undifferentiated; Universities; Walkers; Woman; Work; ZNF145 gene; abstracting; base; career; career development; cell behavior; chromatin immunoprecipitation; cohort; design; embryonic stem cell; experience; in vivo; innovation; insight; interest; male; medical schools; member; mouse model; network models; novel strategies; post-doctoral training; progenitor; programs; promoter; research study; self-renewal; sertoli cell; sperm cell; stem; stem cell biology; stem cell fate; transcription factor

Project Summary/Abstract Spermatogonial stem cells (SSCs) are at the foundation of mammalian spermatogenesis, maintaining sperm production throughout adult life. The molecular mechanisms that control the critical self-renewal vs. differentiation fate decision of SSCs are largely unknown. The hypothesis is that transcription factors primarily restricted to stem and progenitor spermatogonia form regulatory networks to execute gene expression programs important for SSC fate decisions and spermatogenesis. Three Specific Aims will test this hypothesis. Experiments in Specific Aim 1 will employ functional genetics to determine if specific transcription factors are essential for spermatogenesis using a knockdown strategy in ex vivo SSC cultures followed by transplantation to assess stem cell and spermatogenic activity. Specific Aim 2 will identify the target gene repertoire of SSC transcription factors by ChIP-Seq and reveal conserved and unique targets sets. Experiments in Specific Aim 3 will confirm mechanisms of target gene regulation by EMSA and transient transfection transcriptional analysis. These results will form the basis of gene regulatory network models incorporating complex interactions from multiple factors contributing to gene regulation and SSC potential. The proposed studies will be enhance the independent career trajectory of the PI (Brian Hermann) by establishing a new research focus investigating the fundamental regulation of SSCs in the rodent testis under the mentorship of Dr. Kyle Orwig. This is a new research direction for the PI who has established expertise in Sertoli cell gene regulation and primate spermatogenesis and stem cells. The intellectual and technical environment for research in reproductive biology and stem cells at the Magee-Womens Research Institute and University of Pittsburgh is outstanding, all resources required to complete the studies are available, and there is a strong reputation for promoting development of new investigators. This combination of a PI with outstanding potential for independence, an innovative mentor, and superior environment will facilitate all aspects of the proposed studies and advancement of PI's career towards independence. The long-term career goals of the PI are to establish an independent research program in a tenure-track faculty position, make significant scholarly contributions to the understanding of SSCs and spermatogenesis, and develop new approaches for treating male infertility. Three immediate objectives to promote the transition to independence are addressed by the career development award: 1) develop an independent research niche, 2) demonstrate outstanding productivity, and 3) secure additional independent funding. The career development plan involves conceptual mentoring in functional genetics, technical training relating to culture and transplantation of SSCs in the mouse model, instruction in bioinformatics data analysis, informal and structured mentoring in areas relevant to running a successful independent research program, and evaluation of career development progress by an external advisory board. Ultimately, these studies may provide insights about the mechanisms that initiate and maintain spermatogenesis, which has implications for treating male infertility. Investigating the biological properties of SSCs may also expand the understanding of how stem cells behave and contribute to the normal function of a variety of adult tissues. Moreover, additional nurturing of the candidate's career development will promote a competitive, independent research career to contribute substantively to the biomedical sciences.

项目摘要/摘要 精子干细胞（SSC）是哺乳动物的精子发生的基础，维持 精子在整个成人生活中产生。控制关键自我更新VS的分子机制 SSC的分化命运决策在很大程度上未知。假设是转录因子主要是 仅限于STEM和祖细胞精子形成调节网络以执行基因表达 对SSC命运决策和精子发生重要的程序。三个具体目标将检验这一假设。 特定目标1中的实验将采用功能遗传学来确定特定转录因子是否是否 对于在离体SSC培养中使用敲低策略的生理发生至关重要 移植以评估干细胞和精子活性。特定目标2将识别目标基因 通过ChIP-Seq和揭示了保守和独特的目标集的SSC转录因子的曲目。 特定目标3中的实验将确认EMSA和瞬态调节靶基因的机制 转染转录分析。这些结果将构成基因调节网络模型的基础 从多种因素促进基因调节和SSC电位的复杂相互作用。 拟议的研究将增强PI（Brian Hermann）的独立职业轨迹 建立一项新的研究重点，研究啮齿动物睾丸中SSC的基本调节 凯尔·奥维格（Kyle Orwig）博士的指导。对于已经建立了专业知识的PI，这是一个新的研究方向 Sertoli细胞基因调节和灵长类动物的精子发生和干细胞。智力和技术 Magee-Womens研究所的生殖生物学研究和干细胞研究环境 匹兹堡大学非常出色，完成研究所需的所有资源，那里都可以 在促进新研究人员的发展方面享有很高的声誉。 PI与 独立的杰出潜力，创新的导师和卓越的环境将有助于所有人 拟议的研究和PI职业发展独立的各个方面。 PI的长期职业目标是在终身制中建立独立的研究计划 教师职位，对理解SSC和精子发生做出重大的学术贡献， 并开发新的方法来治疗男性不育。促进过渡的三个直接目标 职业发展奖旨在解决独立性：1）开发一个独立的研究利基市场， 2）展示出色的生产力，3）获得额外的独立资金。职业 发展计划涉及功能遗传学的概念指导，与文化有关的技术培训 SSC在鼠标模型中的移植，生物信息学数据分析中的指导，非正式和 与运行成功的独立研究计划相关的领域的结构化指导和评估 外部顾问委员会的职业发展进步。 最终，这些研究可能会提供有关启动和维护机制的见解 精子发生，对治疗男性不育症具有影响。研究的生物学特性 SSC还可以扩展对干细胞行为方式的理解，并促进A的正常功能 各种成年组织。此外，对候选人职业发展的额外培养将促进 竞争性，独立的研究职业为生物医学科学做出实质性贡献。