CAREER: Dynamic transcription factor activity profiling for investigating mechanisms guiding early stage ovarian folliculogenesis

职业：动态转录因子活性分析，用于研究指导早期卵巢卵泡发生的机制

• 批准号: 1552580
• 负责人: Ariella Shikanov
• 金额: $ 50万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1552580&HistoricalAwards=false
• 关键词: CAREER; Dynamic; transcription; factor; activity

PI: Shikanov, Ariella Proposal Number: 1552580Young women and prepubescent girls undergoing toxic treatments for cancer or autoimmune diseases have limited options to preserve their fertility. Fertility preservation is especially challenging in female patients, because females are born with an ovarian reserve comprised of a finite number of follicles that carry immature eggs. Cryopreservation of ovarian tissue prior to toxic treatments and subsequent culture of immature follicles is a promising and novel way to preserve fertility. The challenge is that follicles have low survival and maturation rates in culture, which are attributed to the complex and poorly understood signaling and interactions between the cells in a follicle. The central objective of this CAREER proposal is to discover the key signaling factors involved in the early stage development of ovarian follicles by using systems biology approach. This system will lead to the development of culture conditions for successful follicle culture to be translated to human follicles for fertility preservation, serve as a platform to screen therapeutic compounds, identify pathways with aberrant activity that could be targeted for therapy, or design environments that promote specific cellular processes in regenerative medicine and toxicology. The proposed research will identify numerous transcription factors and establish their role in the growth of early-stage ovarian follicles in vitro. These fundamental studies might provide the foundation for the development of a culture system that can support the development of early stage follicles. In the bench-to-bed translation, this knowledge may be used for fertility preservation in young female cancer survivors suffering from chemotherapy-induced infertility. Our overall research objective is to: (a) establish the dynamic activity of transcription factors during early-stage follicle development in vitro; (b) identify signaling pathways that are responsible for promoting follicle survival and growth through follicle-follicle and follicle-support cells interactions; (c) determine the secretome of cultured early-stage follicles in conditions that allow follicle-follicle and follicle-stroma interactions and correlate the functional paracrine signaling factors with the activity of transcription factors (TFs) to identify the key factors and processes driving successful folliculogenesis. To achieve this objective, we will employ a systems level approach utilizing quantitative experimental characterization with multivariate analysis and modeling. Previous investigator's work has already demonstrated how optimizing biomaterial design can enhance follicle survival rates. We will build upon this system and use the new TRACER technology (TRanscription Activity CEll aRray) to discover the identity, timing and activity pattern of TFs and secreted growth factors involved in folliculogenesis. The key transcription and secreted factors during the development of primary follicles will be investigated in vitro and compared to what is already known for in vivo development. The generated network of transcription factors predicted to be located downstream of the identified secreted factors will describe currently unknown signaling pathway active during early folliculogenesis. The broader research impact of this work will result in culture systems that can maximize ovarian follicle development and provide high quality oocytes for fertility preservation and toxicology screening. This approach has a strong potential to shed light on the mechanism of biological processes of ovarian follicle maturation, which are largely unknown and might benefit basic biologists, bioengineers, clinicians and women facing infertility altogether. The educational outreach program of this proposal integrates middle and high school students with undergraduate and graduate students to the benefit of both groups. The program will incorporate reproductive tissue engineering and biomaterials-related content and projects and will inspire K-12 students to pursue science and engineering careers through conducting K-12 outreach. The investigator is also committed to support women and minority retention in science and engineering through organizational activities and one-on-one mentoring.

主要研究者：Shikanov、Ariella 提案编号：1552580年轻妇女和青春期前女孩接受有毒治疗癌症或自身免疫性疾病有有限的选择，以保持其生育能力。对于女性患者来说，保持生育能力尤其具有挑战性，因为女性生来就具有由有限数量的携带未成熟卵子的卵泡组成的卵巢储备。在毒性治疗之前冷冻保存卵巢组织，随后培养未成熟卵泡是一种有前途的新方法来保持生育能力。挑战在于卵泡在培养中的存活率和成熟率较低，这归因于卵泡中细胞之间复杂且知之甚少的信号传导和相互作用。本CAREER计划的中心目标是通过系统生物学方法发现参与卵泡早期发育的关键信号因子。该系统将导致成功的卵泡培养的培养条件的发展，以转化为人类卵泡的生育力保存，作为一个平台，以筛选治疗化合物，识别具有异常活性的途径，可以靶向治疗，或设计环境，促进再生医学和毒理学中的特定细胞过程。这项研究将确定大量的转录因子，并确定它们在体外早期卵泡生长中的作用。这些基础研究可能为开发支持早期卵泡发育的培养系统提供基础。在工作台到床的翻译，这方面的知识可用于生育能力的保护年轻女性癌症幸存者遭受化疗诱导的不孕症。我们的总体研究目标是：（a）建立体外卵泡发育早期转录因子的动态活性;（B）确定通过卵泡-卵泡和卵泡-支持细胞相互作用促进卵泡存活和生长的信号通路;（c）确定在允许卵泡-卵泡和卵泡-间质相互作用，并将功能性旁分泌信号传导因子与转录因子（TF）的活性相关联，以确定驱动成功卵泡发生的关键因子和过程。为了实现这一目标，我们将采用系统级的方法，利用定量实验表征与多变量分析和建模。以前的研究人员的工作已经证明了优化生物材料设计如何提高卵泡存活率。 我们将建立在这个系统上，并使用新的TRACER技术（转录活性细胞），以发现的身份，时间和活动模式的TF和分泌的生长因子参与卵泡发生。将在体外研究初级卵泡发育过程中的关键转录和分泌因子，并与已知的体内发育进行比较。所产生的网络的转录因子预测位于下游的确定的分泌因子将描述目前未知的信号传导途径，在早期卵泡发育的活动。这项工作的更广泛的研究影响将导致培养系统，可以最大限度地提高卵泡发育，并提供高质量的卵母细胞的生育力保存和毒理学筛选。这种方法具有很强的潜力，可以揭示卵泡成熟的生物过程机制，这在很大程度上是未知的，可能有利于基础生物学家，生物工程师，临床医生和面临不孕症的妇女。这项建议的教育推广方案将初中和高中学生与本科生和研究生结合起来，使这两个群体受益。该计划将纳入生殖组织工程和生物材料相关的内容和项目，并将激励K-12学生通过进行K-12外展来追求科学和工程事业。调查员还致力于通过组织活动和一对一的指导，支持妇女和少数民族保留在科学和工程。