The Role of GCM2 in Parathyroid Gland Homeostasis

GCM2 在甲状旁腺稳态中的作用

• 批准号: 8687644
• 负责人: MICHAEL ALAN LEVINE
• 金额: $ 54.88万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-22 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8687644
• 关键词: Ablation; Adult; Age; Alleles; Atrophic; Bilateral; Binding Sites; Biochemical; Calcium; Cataloging; Catalogs; Cell Death; Cell Line; Cells; Cellular biology; ChIP-seq; Companions; Computer Simulation; DNA-Binding Proteins; Data; Defect; Development; Developmental Biology; Disease of parathyroid glands; Embryo; Embryonic Development; Endoderm Cell; Erinaceidae; Fourth Pharyngeal Pouch; Funding; Future; Gene Dosage; Gene Expression; Genes; Genetic Transcription; Genetically Engineered Mouse; Goals; Growth; Hand; Homeostasis; Human; Hybrids; Hyperparathyroidism; Hyperplasia; Hypoparathyroidism; Knowledge; Lead; Life; Lobe; Location; Medical; Mission; Modeling; Molecular; Molecular Biology; Molecular Genetics; Molecular Target; Mus; Mutate; Neuroglia; Outcome; PTH gene; Parathyroid Neoplasms; Parathyroid gland; Partner in relationship; Pathogenesis; Pathway interactions; Patients; Phosphorus; Physiology; Play; Primordium; Proteins; Public Health; Reagent; Reporter; Repression; Research; Research Project Grants; Role; Serum; Siblings; Signal Transduction; Site; Tamoxifen; Techniques; Technology; Testing; Therapeutic; Thymus Gland; Time; Transcriptional Regulation; Transgenic Mice; United States National Institutes of Health; Work; Yeasts; base; genetic regulatory protein; gland development; innovation; insight; loss of function mutation; mouse model; new therapeutic target; novel; novel strategies; novel therapeutic intervention; overexpression; postnatal; promoter; protein function; public health relevance; recombinase; research study; screening; smoothened signaling pathway; therapeutic target; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): This resubmitted proposal for funding is the result of 2 years of ARRA R01 funding that generated novel data demonstrating a critical role for the parathyroid cell-specific transcription factor Gcm2 (glial cell missing 2) after early embryologica development of the parathyroid glands as well how this protein functions in mature parathyroid glands to control function and survival of parathyroid cells. The long-term goal is use knowledge of the role of Gcm2 to develop novel strategies for medical treatment of patients with hyperparathyroidism. The objective in this application is to identify the extent to which depletion of Gcm2 leads to reduced survival of parathyroid cells in genetically engineered mouse models. The central hypothesis is that expression of Gcm2 in the parathyroid is necessary throughout life to maintain parathyroid cell mass, and that lack of Gcm2 will induce parathyroid cell death. We propose to use mouse models that we have developed to genetically delete the Gcm2 gene conditionally, in a temporally controlled manner, to identify the genes that regulate expression of Gcm2 as well as those that are controlled by Gcm2 action, and to uncover the effect of loss of Gcm2 on mature parathyroid cells. Mice with conditional Gcm2 alleles will also allow us to determine the extent to which ablation of Gcm2 late in life can "rescue" mice that have parathyroid disorders that replicate human hyperparathyroidism. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) determine the role of Sonic hedgehog signaling function and other transcription factor pathway on transcription of the GCM2 gene. Based on our preliminary data, we expect Sonic hedgehog plays a major role in repressing expression of Gcm2 in non-parathyroid cells; and 2) determine the effect of conditional deletion of Gcm2 on the transcriptosome of normal parathyroid glands and on function and size of hyperfunctioning mouse parathyroid glands. We expect that deletion of Gcm2 in normal parathyroid glands will reveal genes that are regulated by Gcm2 and will result in parathyroid atrophy. Moreover, using well-established murine models of primary hyperparathyroidism, we expect that timed deletion of Gcm2 in hyperplastic or adenomatous parathyroid glands will result in decreased survival of parathyroid cells and regression of hyperparathyroidism. The reagents and mouse models are in hand, and the techniques have been established as feasible in the applicants' labs. The approach is innovative because it utilizes novel mouse models, applies new techniques such as enhanced yeast 1-hybrid screening to uncover genes that regulate Gcm2 expression, and uses RNA-seq to develop a comprehensive catalog of the Gcm2-dependent transcriptosome. The proposed research is significant because it is expected to advance our understanding of parathyroid cell biology, and ultimately, to identify molecular targets that will allow development of new medical treatments for primary and tertiary hyperparathyroidism in humans.

DESCRIPTION (provided by applicant): This resubmitted proposal for funding is the result of 2 years of ARRA R01 funding that generated novel data demonstrating a critical role for the parathyroid cell-specific transcription factor Gcm2 (glial cell missing 2) after early embryologica development of the parathyroid glands as well how this protein functions in mature parathyroid glands to control function and survival of parathyroid cells.长期目标是利用GCM2作用的知识来制定甲状旁腺功能亢进症患者的医疗治疗新颖策略。本应用程序的目的是确定耗尽的程度 GCM2的GCM2导致基因工程小鼠模型中甲状旁腺细胞的存活率降低。中心假设是，GCM2在甲状旁腺中的表达在整个生命中都是维持甲状旁腺细胞质量的必要条件，并且缺乏GCM2会诱导甲状旁腺细胞死亡。我们建议使用已开发的小鼠模型以临时控制的方式有条件地删除GCM2基因，以识别调节表达表达的基因 GCM2以及由GCM2作用控制的GCM2，并发现GCM2损失对成熟甲状旁腺细胞的影响。具有条件GCM2等位基因的小鼠还将使我们能够确定生命后期GCM2的消融程度可以“拯救”具有复制人类甲状旁腺功能亢进症的甲状旁腺功能障碍的小鼠。在强大的初步数据的指导下，将通过追求两个具体目标来检验该假设：1）确定声波刺猬信号传导函数和其他转录因子途径在GCM2基因转录中的作用。根据我们的初步数据，我们期望声波刺猬在抑制非甲状旁腺细胞中GCM2的表达中起着重要作用。 2）确定GCM2条件缺失对正常甲状旁腺的转录体以及对小鼠甲状旁腺的功能和大小的影响。我们预计在正常甲状旁腺中GCM2的缺失将揭示受GCM2调节的基因，并会导致甲状旁腺萎缩。此外，使用原发性甲状旁腺功能亢进的鼠模型，我们预计GCM2在增生或腺瘤甲状旁腺中的定时缺失将导致甲状旁腺细胞的存活率降低和甲状腺功能亢进的降低。试剂和小鼠模型已经掌握，并且在申请人实验室中已确定了这些技术。该方法具有创新性，因为它利用了新型的小鼠模型，应用了新技术，例如增强的酵母1-杂交筛选，以发现调节GCM2表达的基因，并使用RNA-Seq来开发GCM2依赖性转录体的全面目录。拟议的研究之所以重要，是因为有望提高我们对甲状旁腺细胞生物学的理解，并最终确定将允许为人类开发原发性和高甲状腺功能亢进症的新药物治疗的分子靶标。