Wnt-10A and Thyrotrope Cells

Wnt-10A 和促甲状腺激素细胞

• 批准号: 7885255
• 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 2011-12-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7885255
• 关键词: Applications Grants; Area; Basic Science; Biological Models; Brain-Derived Neurotrophic Factor; Cell Proliferation; Cell model; Cell physiology; Cells; Complementary DNA; Data; Elements; Embryonic Development; Endocrinology; Feedback; Gene Expression; Genes; Genetic Transcription; Goals; Grant; Growth; Human Development; Hyperplasia; In Situ Hybridization; Ligands; Malignant Neoplasms; Mediating; Mediator of activation protein; Medicine; Messenger RNA; Microarray Analysis; Modeling; Molecular Biology; Molecular Profiling; Morphogenesis; Mus; Physicians; Pituitary Gland; Play; Postdoctoral Fellow; Process; Production; Public Health; Regulation; Regulatory Element; Research; Research Methodology; Research Personnel; Research Proposals; Role; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Small Interfering RNA; Somatostatin; Technology; Thyroid Hormones; Thyrotrope Cell; Thyrotropin-Releasing Hormone; Thyroxine; Training; Transcript; Transcriptional Regulation; Transgenic Mice; alpha Subunit Glycoprotein Hormones; career; cdc Genes; cell growth; cis acting element; in vivo; knock-down; loss of function; mouse model; pituitary gland development; promoter; receptor; research study; skills; tumor

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）： 正常和异常垂体功能中 Wnt 信号传导的基础科学是内分泌学研究的一个令人兴奋的领域。本研究计划的目标是表征 Wnt-10A 对促甲状腺细胞的影响。利用鼠 TtT-97 促甲状腺肿瘤增生模型，我之前证明用甲状腺激素治疗 TtT-97 肿瘤可减小肿瘤大小，并与 Wnt-10A 配体表达减少相关。本拨款提案的目的是阐明 Wnt-10A 在促甲状腺素中的作用，具体而言：1) 表征 Wnt-10A 对 Wnt 信号传导、细胞周期基因和促甲状腺素细胞生长的影响，2) 鉴定调节 TtT-97 细胞中 Wnt-10A 表达的顺式作用启动子元件，3) 建立垂体定位和甲状腺激素反应性 体内 Wnt-10A 表达的影响，以及 4) 阐明 Wnt-10A 过表达对促甲状腺细胞的影响。为了更好地表征调节 Wnt-10A 基因表达的因素，将比较受调节 (TtT-97) 和不受调节 (α-TSH) 促甲状腺素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。具体来说，siRNA技术将用于研究Wnt-10A敲低对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过表达研究，以表征Wnt信号通路和Wnt-10A激活的细胞周期基因，并进一步确定重要的调控元件。然后提出进一步的研究来定义调节基础活性和甲状腺激素反应性的 Wnt-10A 启动子的顺式作用元件。为了确定 Wnt-10A 的促甲状腺素定位及其在小鼠垂体中的甲状腺激素反应性，随后提出了原位杂交研究。最后，为了确定 Wnt-10A 过度表达对促甲状腺素细胞的体内影响，提出了转基因小鼠模型。 这项基础研究的主要目标是增进对促甲状腺素细胞生理学的理解。首席研究员 Janice Kerr 是一名医生、博士后研究员，致力于基础科学研究和学术医学事业。主要研究员的短期目标是提高她在分子生物学和 Wnt 信号传导领域的研究技能。她的长期目标是成为内分泌领域训练有素的独立科学家。 拟议的研究与公共健康相关，因为 Wnt 信号传导是正常发育和人类恶性肿瘤中公认的因素，对 Wnt-10A 的进一步了解可能会阐明其在正常和异常垂体过程中的作用。