Wnt-10A and Thyrotrope Cells

Wnt-10A 和促甲状腺激素细胞

• 批准号: 7093800
• 负责人: JANICE M KERR
• 金额: $ 13.19万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7093800
• 关键词: RNA interference; Wnt gene /protein; biological signal transduction; cell cycle; cell proliferation; cytogenetics; disease /disorder model; endocrinology; gene deletion mutation; gene expression; genetic promoter element; genetic transcription; genetically modified animals; hormone related neoplasm /cancer; hyperplasia; in situ hybridization; laboratory mouse; pituitary gland; protein localization; protein structure function; thyroid hormones

DESCRIPTION (provided by candidate): The basic science of Wnt signaling in normal and abnormal pituitary gland function is an area of exciting endocrinology research. The goal of this research proposal is to characterize the effects of Wnt-10A on thyrotrope cells. Utilizing the murine TtT-97 thyrotropic tumor model of hyperplasia, I previously demonstrated that treatment of TtT-97 tumors with thyroid hormone reduced tumor size in association with decreased expression of the Wnt-10A ligand. The purpose of this grant proposal is to elucidate the role of Wnt-10A in thyrotropes, specifically: 1) to characterize the effects of Wnt-10A on Wnt signaling, cellcycle genes, and growth in thyrotrope cells, 2) to identify the cis-acting promoter elements that regulate Wnt-10A expression in TtT-97 cells, 3) to establish the pituitary localization and thyroid hormone responsiveness of Wnt-10A expression, in vivo, and 4) to elucidate the effects of Wnt-10A over-expression on thyrotrope cells. To better characterize factors that regulated Wnt-10A gene expression, model systems of regulated (TtT-97) and unregulated (alpha-TSH) thyrotrope cell growth will be compared. In order to achieve these scientific objectives, research methods that utilize Wnt-10A loss-of-function and over-expression will be performed. Specifically, siRNA technology will be utilized to study the effects of Wnt-10A knock-down on cell-cycle genes and growth in TtT-97 and alpha-TSH cells. Next, Wnt-10A over- expression studies will then be performed to characterize the Wnt signaling pathway and cell-cycle genes that are activated by Wnt-10A and further define important regulatory elements. Further studies are then proposed to define the cis-acting elements of the Wnt-10A promoter that regulate basal activity and thyroid-hormone responsiveness. To establish the thyrotrope localization of Wnt-10A and its thyroid hormone responsiveness in the murine pituitary gland, in situ hybridization studies are then proposed. Lastly, to determine the in vivo effects of Wnt-10A over-expression on thyrotrope cells, a transgenic mouse model is proposed. The primary goal of this basic research study is to advance the understanding of thyrotrope cell physiology. The principle investigator, Janice Kerr, is a physician, post-doctoral fellow with a strong commitment to basic science research and a career in academic medicine. The short-term goals of the principle investigator are to advance her research skills in the area of molecular biology and Wnt signaling. Her long-term goals are to become a well-trained, independent-scientist in the field of endocrinology. The research proposed is relevant to public health in that Wnt signaling is a well-recognized factor in normal development and human malignancies, and further understanding of Wnt-10A may elucidate its role in normal and abnormal pituitary processes.

描述(由候选人提供)： Wnt信号在正常和异常垂体腺功能中的基础科学是内分泌学研究的热点。这项研究计划的目的是表征WNT-10A对促甲状腺细胞的影响。利用小鼠TTT-97促甲状腺肿瘤增殖模型，我以前证明了用甲状腺激素治疗TTT-97肿瘤缩小了肿瘤大小，与WNT-10A配体的表达减少有关。本拨款建议的目的是阐明WNT-10A在促甲状腺激素中的作用，特别是：1)表征WNT-10A对WNT信号、细胞周期基因和甲状腺细胞生长的影响，2)确定在TTT-97细胞中调节WNT-10A表达的顺式作用启动子元件，3)在体内建立WNT-10A表达的垂体定位和甲状腺激素反应性，以及4)阐明WNT-10A过表达对甲状腺细胞的影响。为了更好地表征调节Wnt-10A基因表达的因素，我们将比较调节(TTT-97)和非调节(α-TSH)促甲状腺细胞生长的模型系统。 为了实现这些科学目标，将进行利用WNT-10A功能丧失和过度表达的研究方法。具体地说，将利用siRNA技术研究Wnt-10A基因敲除对TTT-97和α-TSH细胞的细胞周期基因和生长的影响。接下来，将进行WNT-10A过度表达的研究，以确定由WNT-10A激活的WNT信号通路和细胞周期基因，并进一步确定重要的调控元件。然后建议进行进一步的研究，以确定WNT-10A启动子调节基础活性和甲状腺激素反应性的顺式作用元件。为了确定WNT-10A在小鼠脑下垂体中的促甲状腺功能定位及其对甲状腺激素的反应性，提出了原位杂交研究。最后，为了确定Wnt-10A在体内过表达对促甲状腺细胞的影响，我们建立了转基因小鼠模型。 这项基础研究的主要目标是促进对促甲状腺细胞生理学的了解。首席研究员贾尼斯·科尔是一名内科医生，博士后，对基础科学研究和学术医学事业有着坚定的承诺。这位首席研究员的短期目标是提高她在分子生物学和Wnt信号领域的研究技能。她的长期目标是成为内分泌学领域训练有素的独立科学家。 这项研究与公众健康有关，因为Wnt信号在正常发育和人类恶性肿瘤中是一个公认的因子，对Wnt-10A的进一步了解可能会阐明它在正常和异常垂体过程中的作用。