Murine Knockout Model of Mevalonic Aciduria

甲羟戊酸尿症小鼠敲除模型

基本信息

批准号：
7938235
负责人：
K Michael GIBSON
金额：
$ 4.82万
依托单位：
MICHIGAN TECHNOLOGICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7938235
关键词：
3-hydroxy-3-methylglutaryl-coenzyme A 7-dehydrocholesterol Abbreviations Ablation Acetyl Coenzyme A Age Amino Acids Animal Husbandry Animals Bile Acids Cell physiology Cerebellar Ataxia Cholesterol Code Coenzyme A Commit Complex Data Defect Diphosphates Disease Dolichol Embryo Erinaceidae Evaluation Fetal Development Figs - dietary Functional disorder Gender Gene Deletion Gene Mutation Gene Targeting Genes Gestational Age Hematopoiesis Heme Hereditary Disease Human Hydroxymethylglutaryl-CoA reductase Hyperimmunoglobulin Syndrome Hyperimmunoglobulinemia D IgG3 Immune System Diseases Immunoglobulin A Immunoglobulin D Immunoglobulins Inflammatory Interferons Interleukin-6 Isoprene Knock-in Mouse Knock-out Lymphocyte Mammalian Cell Methodology Methods Mevalonate kinase Mevalonic Acid Missense Mutation Modeling Mus Neurologic Pathway interactions Patients Phenotype Protein Isoprenylation Proteins Research Personnel Residual state Role Serum Site Squalene Staging Syndrome TNF gene Time Transfer RNA Ubiquinone Vitamin D Work acetoacetyl CoA base cohort cytokine design embryonic stem cell enzyme activity enzyme deficiency implantation inorganic phosphate knock-down membrane biogenesis mevalonate steroid hormone tool urinary

项目摘要

DESCRIPTION (from the application): Mevalonate kinase (MvK) catalyzes the first committed step in cholesterol and isoprene synthesis, and is the site of two human enzyme deficiencies, severe mevalonic aciduria (MA) and hyperimmunoglobulin D syndrome (HIDS), both manifesting autoinflammatory disease. The investigators' long-term objective is to understand the pleiotropic pathophysiology associated with MA and HIDS, and the potential role of MvK in diverse cellular processes. To work toward this objective, they have utilized a gene-trap approach to ablate MvK in the mouse. Whereas MvK-/- mice appear to die embryonically, MvK+/- mice survive and manifest a hyperinflammatory phenotype reminiscent of that seen in both HIDS and MA patients, but absent the additional neurological sequelae observed in MA patients. The specific aim is to identify the embryological age of fetal loss for MvK-/- mice using the investigators' current gene-trap construct, assess structural anomalies that might be present and determine whether implantation occurs, and then generate a viable MvK-/- mouse via knock-in of specific human MvK missense mutations. The rationale is that HIDS patients manifest higher residual MvK enzyme activity (1.4-5.7% of control) than MA patients (0-0.2% of control). Thus, the investigators speculate that MvK+/- animals (with ~50% of MvK+/+ enzyme activity) might more readily recapitulate the HIDS phenotype, whereas it will require greater knock-down of murine MvK activity to recapitulate the human MA phenotype, but not total ablation via gene deletion. The hypothesis is that knock-in of specific human MvK missense mutations will produce a viable MvK-/- mouse that recapitulates the neurological phenotype and immune disease observed in MA patients. The investigators will adhere to a cohort control design, with age- and gender-matched controls for all studies, and will employ standard methods of animal husbandry, embryo isolation and characterization, and gene targeting. MvK+/- and MvK-/- mice will help unravel pathophysiology associated with MA and HIDS, while providing tools for characterization of other autoinflammatory disorders as well as monogenic/complex genetic diseases in which cholesterol pathway function has been implicated.

描述（来自申请）：甲羟戊酸激酶 (MvK) 催化胆固醇和异戊二烯合成的第一个关键步骤，并且是两种人类酶缺乏症、严重甲羟戊酸尿症 (MA) 和高免疫球蛋白 D 综合征 (HIDS) 的发生部位，两者均表现为自身炎症疾病。研究人员的长期目标是了解与 MA 和 HIDS 相关的多效性病理生理学，以及 MvK 在不同细胞过程中的潜在作用。为了实现这一目标，他们利用基因陷阱方法消除小鼠中的 MvK。 MvK-/- 小鼠似乎在胚胎时死亡，而 MvK+/- 小鼠则存活并表现出与 HIDS 和 MA 患者相似的高炎症表型，但没有在 MA 患者中观察到的额外神经系统后遗症。具体目标是使用研究人员当前的基因陷阱构建来确定 MvK-/- 小鼠胎儿丢失的胚胎年龄，评估可能存在的结构异常并确定是否发生植入，然后生成可行的 MvK-/- 小鼠小鼠通过敲入特定的人类 MvK 错义突变。其基本原理是，HIDS 患者表现出比 MA 患者（对照的 0-0.2%）更高的残余 MvK 酶活性（对照的 1.4-5.7%）。因此，研究人员推测 MvK+/- 动物（具有约 50% 的 MvK+/+ 酶活性）可能更容易重现 HIDS 表型，而需要更大程度地敲低小鼠 MvK 活性才能重现人类 MA 表型，但是不是通过基因删除来完全消除。假设敲入特定的人类 MvK 错义突变将产生可存活的 MvK-/- 小鼠，它重现了 MA 患者中观察到的神经表型和免疫疾病。研究人员将坚持队列对照设计，对所有研究进行年龄和性别匹配的对照，并将采用畜牧业、胚胎分离和表征以及基因靶向的标准方法。 MvK+/- 和 MvK-/- 小鼠将有助于揭示与 MA 和 HIDS 相关的病理生理学，同时提供表征其他自身炎症性疾病以及涉及胆固醇途径功能的单基因/复杂遗传疾病的工具。