Creation of a conditional IL-6 knockout mouse

条件性IL-6基因敲除小鼠的创建

• 批准号: 7480242
• 负责人: CLIFFORD Scott DEUTSCHMAN
• 金额: $ 19.31万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7480242
• 关键词: Acute; Adenovirus Vector; Affect; Animals; Antibodies; Arts; Blood Circulation; Breeding; Caring; Cause of Death; Cells; Characteristics; Cleaved cell; Communication; Complement Activation; Conceptions; Critical Illness; Development; Disease; Enzymes; Exons; Exploratory/Developmental Grant; Functional disorder; Generations; Genes; Germ; Grant; Heart; Hepatic; IL6 gene; Immune; Individual; Inflammation; Inflammatory; Interleukin-6; Investigation; Knock-out; Knockout Mice; Leukocytes; Ligation; Liver; Lung; Mediating; Mediator of activation protein; Modeling; Morbidity - disease rate; Multiple Organ Failure; Mus; Normal Cell; Organ; Outcome; Pathogenesis; Pathologic; Pathway interactions; Patients; Process; Production; Public Health; Puncture procedure; Relative (related person); Role; Sepsis; Signal Transduction; Site; Syndrome; System; Techniques; Testing; Time; Tissues; Trachea; United States; attenuation; body system; cytokine; immune depression; improved; mortality; novel; recombinase; response; septic; tool

DESCRIPTION (provided by applicant): Sepsis is a major cause of mortality and morbidity in the critically ill. We have been studying the disorder using a murine model (cecal ligation and puncture or CLP) for some time and have uncovered some important abnormalities in several organ systems, notably liver, lung and heart. Key among our findings is a loss in the activity of an important inflammatory mediator, the cytokine interleukin (IL) - 6. We have extended our investigations to mice with a congenital absence of IL-6 (IL-6 -/- or "knockout" mice). All the abnormalities we observed in wild type (IL-6 +/+) mice were worse in IL-6 -/- animals. However, in IL-6 -/- mice, the knockout is generated at conception. Therefore, compensatory pathways can develop. In addition, the loss of IL-6 in IL-6 -/- mice does not truly mimic what occurs in sepsis in wild types. The ability to acutely eliminate IL- 6 would be extremely useful in studying sepsis. Unfortunately, current techniques to eliminate IL-6 have significant drawbacks. Use of IL-6 -/- mice is limited by an inability to extrapolate findings in animals with a congenital, in-born error to the acute situation. Use of antibodies directed at IL-6 is effective in eliminating IL-6 from the circulation but not from tissues, where most of the damage in sepsis occurs. In this grant, we propose another approach. We will generate and breed a mouse from which IL-6 can be acutely eliminated. This will be accomplished by creating an IL-6 construct gene that is flanked by two sites that can be cleaved by an enzyme called cre-recombinase. This construct will be introduced into germ tissue and a mouse containing this sequence will be bred. Once we have established a line of animals, we will be able to eliminate IL-6 from lung and liver by administering an adenoviral vector expressing cre-recombinase into the trachea or the bloodstream. This will allow acute elimination of IL-6 from lung and liver. The availability of such a tool will allow us to better investigate that role of IL-6 in the pathogenesis of sepsis-induced changes in these organs. This will provide increased understanding op septic pathophysiology. Sepsis is an important cause of mortality and morbidity. It has been estimated that in excess of $16 billion are spent each year in the use to treat this disorder. As such, sepsis constitutes a major public health problem. . Unfortunately, the pathophysiology of the disease is poorly understood. A better understanding of this disease is extremely relevant to the care of the citizens of the United States.

描述(由申请人提供)：败血症是危重病人死亡和发病的主要原因。我们使用小鼠模型(盲肠结扎和穿孔或CLP)研究这种疾病已经有一段时间了，并发现了几个器官系统的一些重要异常，特别是肝脏、肺和心脏。我们发现的关键是一种重要的炎症介质--细胞因子IL-6活性的丧失。我们将我们的研究扩展到先天缺乏IL-6的小鼠(IL-6-/-或“基因敲除”小鼠)。我们在野生型(IL-6+/+)小鼠中观察到的所有异常在IL-6-/-动物中更严重。然而，在IL-6-/-小鼠中，基因敲除是在受孕时产生的。因此，补偿途径是可以发展的。此外，IL-6-/-小鼠中IL-6的丢失并不能真正模拟野生型败血症时的情况。准确清除IL-6的能力在研究脓毒症方面将是非常有用的。不幸的是，目前消除IL-6的技术存在重大缺陷。IL-6-/-小鼠的使用受到限制，因为无法推断患有先天、先天错误的动物的研究结果对急性情况的影响。使用针对IL-6的抗体可以有效地从循环中清除IL-6，但不能从组织中清除IL-6，而组织是脓毒症中大多数损害发生的地方。在这项拨款中，我们提出了另一种方法。我们将培育出一种可以迅速消除IL-6的小鼠。这将通过创建一个IL-6构建基因来实现，该基因两侧有两个可被一种名为cre-重组酶的酶切割的位点。这种结构将被引入生殖细胞组织中，并将培育出含有该序列的小鼠。一旦我们建立了一系列动物，我们将能够通过将表达重组酶的腺病毒载体注射到气管或血液中来消除肺和肝脏中的IL-6。这将使IL-6从肺和肝脏中迅速消除。这种工具的出现将使我们能够更好地研究IL-6在脓毒症引起的这些器官改变的发病机制中的作用。这将增加对败血症病理生理学的了解。脓毒症是导致死亡和发病的重要原因。据估计，每年用于治疗这种疾病的资金超过160亿美元。因此，败血症构成了一个重大的公共卫生问题。。不幸的是，人们对这种疾病的病理生理学知之甚少。更好地了解这种疾病与美国公民的护理极其相关。