A novel mouse model to monitor inflammasome activation in vivo

 描述（由申请人提供）：炎性体活性对于体内平衡是必不可少的，但是受损的和过度的活性导致广谱的人类炎性疾病，因此在体内疾病进展期间确定炎性体活性对于推进我们对潜在疾病病理学的认识和开发新的疗法是重要的。然而，目前还不存在监测和量化体内炎性小体活性的方法，甚至离体和体外测量需要长时间操纵细胞，这对动力学研究产生不利影响，并且还需要昂贵的底物。我们开发了一种新的双报告小鼠模型，允许基于发光和荧光的体内和体外检测和定量炎性小体活性，而无需任何操作。在目标1中，我们建议在体外和体内表征这种新模型，确定其特异性并将其与当前最先进的方法进行对比。在目标2中，我们提出使用两种复杂的炎性小体依赖性疾病，即Cryopyrinopathies（CAPS）和结肠炎，为这种新模型提供原理证明。我们预计这些研究最终将通过在生理学和病理学过程中对炎性小体活性进行新的、更相关的研究来积极影响人类健康，并进一步成为研究炎性小体靶向治疗的临床前研究的非常有用的模型。

 DESCRIPTION (provided by applicant): Inflammasome activity is essential for homeostasis, but impaired and excessive activity causes a wide spectrum of human inflammatory disease, and therefore determining inflammasome activity during disease progression in vivo is important to advance our knowledge on the underlying disease pathologies and to develop novel therapies. Currently, however, no approaches exist to monitor and quantify inflammasome activity in vivo, and even ex vivo and in vitro measurements require prolonged manipulation of cells adverse affecting kinetic studies and further, require expensive substrates. We developed a novel dual reporter mouse model allowing luminescent and fluorescent-based in vivo and ex vivo detection and quantification of inflammasome activity without any manipulation. In aim 1 we propose to characterize this novel model in vitro and in vivo, determine its specificity and contrast it with currently state of the art methods. In aim 2 we propose to provide proof-of-principle for this novel model using two complex inflammasome-dependent diseases, namely Cryopyrinopathies (CAPS) and colitis. We expect these studies will ultimately positively affect human health by enabling novel, more relevant studies of inflammasome activity during physiology and pathology and further will be a very useful model for pre-clinical studies investigating inflammasome-targeted therapies.