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）：败血症是重病死亡率和发病率的主要原因。我们一直在使用鼠模型（Cecal连接和穿刺或CLP）研究该疾病，并发现了几种器官系统，尤其是肝脏，肺和心脏的一些重要异常。我们发现的关键是重要的炎症介质的活性损失，细胞因子白介素（IL）-6。我们已经将研究扩展到没有先天性IL-6（IL-6 - / - 或“敲除”小鼠的小鼠的小鼠。在IL-6 - / - 动物中，我们在野生型（IL-6 +/ +）小鼠中观察到的所有异常都较差。但是，在IL-6 - / - 小鼠中，敲除在受孕时产生。因此，可以发展补偿途径。另外，IL-6 - / - 小鼠中IL-6的损失并不能真正模仿野生型败血症中发生的情况。急性消除IL-6的能力在研究败血症方面非常有用。不幸的是，消除IL-6的当前技术具有明显的缺点。使用IL-6 - / - 小鼠的使用受到无法推断出具有先天性，内向误差的动物的发现的限制。使用针对IL-6的抗体的使用有效地消除了从循环中消除IL-6，而不是从组织中消除IL-6，在败血症中大部分损害发生。在这笔赠款中，我们提出了另一种方法。我们将生成并繁殖一只小鼠，可以从中急剧消除IL-6。这将通过创建一个IL-6构造基因来实现，该基因的两侧是两个可以被称为Cre-recombinase酶裂解的位点。该构建体将被引入细菌组织，并将繁殖包含该序列的小鼠。一旦我们建立了一系列动物，我们将能够通过施用表达Cre-cobibinase的腺病毒载体从气管或血液中消除肝脏中的IL-6。这将允许从肺和肝脏中急性消除IL-6。这种工具的可用性将使我们能够更好地研究IL-6在败血症诱导的这些器官变化的发病机理中的作用。这将提供更多的理解性化味病理生理学。败血症是死亡率和发病率的重要原因。据估计，每年用于治疗这种疾病的每年超过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，该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照顾非常相关。