Development of GM-CSF reporter and reporter/fate-reporter mice

GM-CSF记者和记者/命运记者小鼠的开发

• 批准号: 8582816
• 负责人: Bogoljub Ciric
• 金额: $ 18.43万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8582816
• 关键词: Address; Area; Autoimmune Process; Autoimmunity; Biological; Biology; Biomedical Research; Cells; Clinical Trials; Colony-Stimulating Factor Gene; Communicable Diseases; Cytokine Gene; Development; Disease; Elements; Environment; Fetal Development; Gene Expression; Generations; Genetic; Genetic Transcription; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoiesis; Immune system; Immunity; Immunology; Individual; Inflammation; Inflammatory; Knockout Mice; Lead; Life; Location; Longevity; Malignant Neoplasms; Modification; Monitor; Mus; Phenotype; Physiological; Physiological Processes; Placentation; Process; Production; Proteins; PubMed; Publishing; Reporter; Reporter Genes; Reporting; Research; Role; Stem cells; Testing; Transgenes; Transgenic Mice; Transgenic Organisms; Wound Healing; cytokine; design; innovation; insight; interest; mouse Cre recombinase; novel; public health relevance; receptor; recombinase; stem; therapeutic target; tool; trafficking; vaccine development

DESCRIPTION (provided by applicant): Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been studied for over forty years, and is among the most extensively researched cytokines. GM-CSF is involved in a variety of physiological processes ranging from wound healing and haematopoiesis to placental and fetal development, but it is best known for its role as a pro-inflammatory cytokine in immunity and inflammation. GM-CSF biology has been studied in various branches of immunology, such as autoimmunity, cancer, infectious diseases and vaccine development. In the immune system, GM-CSF has essential and non-redundant roles that make it an attractive therapeutic target. Several clinical trials targeting GM-CSF are underway, and initiation of additional trials is being considered. Both GM-CSF and its receptor knockout mice have been generated and have provided important insights into functioning of the immune system. However, transgenic mice that report GM-CSF expression do not exist, and Specific Aim 1 of this proposal is to generate such mice. We have designed genetic modifications in the GM-CSF reporter (Gr) mice that will readily enable subsequent generation of GM-CSF reporter/fate-reporter (Gr/fr) mice by crossing with existent transgenic mice, and this constitutes Specific Aim 2. The use of fate-reporter mice, which report gene expression even after expression ceases, has proved to be a valuable approach for studying questions that could not be adequately addressed otherwise. The principal conceptual innovation in the transgenic mice that we want to generate is the presence of two reporter genes, one that reports ongoing GM-CSF expression, the other reporting that GM-CSF has been expressed. This enables easy differentiation between current and past expression, which is not the case with fate-reporter mouse lines now being used. The Gr/fr mice will have normal GM-CSF expression, allowing studies of its biology in a physiological context. In Specific Aim 3 we will address a relevant biological question that cannot be addressed without Gr/fr mice, exemplifying advances that can be made by use of these mice. Even though GM-CSF production is crucial for function of Th cells, a majority of them do not produce GM-CSF. It is not known whether subpopulations of Th cells have stable or transient phenotype with regard to GM-CSF expression, nor is it known which factors (a) determine Th cell expression of GM-CSF or (b) change its current GM-CSF- expressing status. We will test the hypothesis that subpopulations of Th cells have stable GM-CSF-expressing or -non-expressing phenotype. We expect that upon completion of this project we will have generated and comprehensively characterized Gr and Gr/fr mouse lines. We will also use Gr/fr mice to answer important biological questions, which will demonstrate their capacity to facilitate advances that would not be feasible otherwise.

描述（由申请人提供）：粒细胞-巨噬细胞集落刺激因子（GM-CSF）已经研究了四十多年，是研究最广泛的细胞因子之一。GM-CSF参与从伤口愈合和造血到胎盘和胎儿发育的各种生理过程，但它最为人所知的是其在免疫和炎症中作为促炎细胞因子的作用。GM-CSF生物学已经在免疫学的各个分支中进行了研究，例如自身免疫、癌症、传染病和疫苗开发。在免疫系统中，GM-CSF具有重要和非冗余的作用，使其成为有吸引力的治疗靶点。目前正在进行几项针对GM-CSF的临床试验，并正在考虑启动其他试验。 已经产生了GM-CSF及其受体敲除小鼠，并提供了对免疫系统功能的重要见解。然而，报告GM-CSF表达的转基因小鼠并不存在，本提案的具体目标1是产生这样的小鼠。我们已经在GM-CSF报告基因（Gr）小鼠中设计了遗传修饰，通过与现有转基因小鼠杂交，可以很容易地产生GM-CSF报告基因/fate-报告基因（Gr/fr）小鼠，这构成了特异性目标2。使用命运报告小鼠，报告基因表达，即使在表达停止后，已被证明是一个有价值的方法来研究的问题，不能充分解决，否则。 我们想要产生的转基因小鼠的主要概念创新是存在两个报告基因，一个报告正在进行的GM-CSF表达，另一个报告GM-CSF已经表达。这使得容易区分当前和过去的表达，这不是现在使用的命运报告小鼠系的情况。Gr/fr小鼠将具有正常的GM-CSF表达，允许在生理背景下研究其生物学。 在具体目标3中，我们将解决一个相关的生物学问题，如果没有Gr/fr小鼠，就无法解决这个问题，并举例说明使用这些小鼠可以取得的进展。尽管GM-CSF的产生对Th细胞的功能至关重要，但它们中的大多数不产生GM-CSF。目前还不知道Th细胞亚群在GM-CSF表达方面是否具有稳定或瞬时表型，也不知道哪些因子（a）决定Th细胞表达GM-CSF或（B）改变其当前的GM-CSF表达状态。我们将检验Th细胞亚群具有稳定的GM-CSF表达或不表达表型的假设。 我们预计，在该项目完成后，我们将产生和全面表征Gr和Gr/fr小鼠品系。我们还将使用Gr/fr小鼠来回答重要的生物学问题，这将证明它们有能力促进否则不可行的进步。