A novel mouse model to monitor inflammasome activation in vivo

 描述(申请人提供)：炎症小体活动对体内平衡是必不可少的，但活动受损和过度会导致广泛的人类炎症性疾病，因此测定体内疾病进展过程中的炎症小体活动对于增进我们对潜在疾病病理的了解和开发新的治疗方法是重要的。然而，目前还没有监测和量化体内炎性小体活性的方法，即使是体外和体外测量也需要对细胞进行长时间的操作，从而影响动力学研究，而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症体活动，而无需任何操作。在目标1中，我们建议在体外和体内对这一新模型进行表征，确定其特异性，并与当前最先进的方法进行比较。在目标2中，我们建议使用两种复杂的炎症性依赖疾病，即冷冻症(CAPS)和结肠炎，为这一新模型提供原理证明。我们预计，这些研究最终将对人类健康产生积极影响，因为它们能够对生理学和病理学过程中的炎症体活动进行新的、更相关的研究，并进一步成为研究炎症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.