Investigation into Mechanisms NLRP3 Activation by Nickel Associated Multi-Walled

描述（由申请人提供）：在动物模型中，某些工程纳米材料（ENM）已被证明会引起显著的肺部病理变化，这引起了人们对随着使用和暴露的增加，人类健康影响将出现的担忧。然而，基于ENM的物理和表面特性的机械预测模型尚未建立，以帮助保护人类健康。我们实验室的其他工作表明，NLRP3炎性体（NLRP3）的体外激活与体内炎症之间存在关联。在初步数据中，我们已经证明镍相关MWCNT （Ni-MWCNT）在体外通过原代肺泡巨噬细胞（AM）或THP-1细胞激活NLRP3的能力可能与肺部炎症和病理有关。因此，我们认为NLRP3的体外激活似乎是肺部炎症的可靠预测因子。区分生物活性和良性ENM的基础很可能与ENM被吞噬的能力和/或破坏溶酶体导致组织蛋白酶B释放的能力有关。我们假设ENM的体内炎症电位与溶酶体破坏活性和NLRP3激活密切相关。根据初步数据，我们提出MWCNT的炎症潜力将取决于伴生镍的数量。因此，我们的目标是：1。在小鼠暴露于明确且特征明确的Ni-MWCNT后，确定AM中NLRP3炎性体活性与肺部病理之间的关系。2. 利用THP-1细胞确定Ni-MWCNT表面性质（镍的量和表面电荷）对引起细胞毒性和NLRP3炎性体活化程度的重要性。最后，我们将确定MWCNT-Ni的内化，随后的溶酶体膜破坏和组织蛋白酶B的释放是NLRP3炎症小体激活的起始事件。因此，这项研究的信息对于确定安全ENM的特征和建立作用机制非常重要，并可能导致使用THP-1细胞的体外筛选平台。

DESCRIPTION (provided by applicant): Certain engineered nanomaterials (ENM) have been shown to cause significant lung pathological changes in animal models raising concern that human health effects will emerge with increasing use and exposure. However, a mechanistic predictive model based on physical and surface properties of ENM has not been established to aid in protecting human health. Other work in our laboratory has shown an association between in vitro NLRP3 inflammasome (NLRP3) activation and in vivo inflammation. In preliminary data, we have shown that there may be a correlation between the ability of nickel associated MWCNT (Ni-MWCNT) to activate NLRP3 in vitro using primary alveolar macrophages (AM) or THP-1 cells with lung inflammation and pathology. Therefore, we propose that in vitro activation of NLRP3 appears to be a reliable predictor of lung inflammation. The basis for the distinction between bioactive and benign ENM is most likely associated with the ability of ENM to be phagocytosed and/or ability to disrupt lysosomes causing cathepsin B release. We hypothesize that the in vivo inflammatory potential of ENM correlates well with lysosomal disrupting activity and NLRP3 activation. Based on preliminary data we propose that the inflammatory potential of MWCNT will be dependent on the amount of associated nickel. Therefore, our aims are: 1. Determine the relationship between NLRP3 inflammasome activity in AM and lung pathology in mice following exposure to defined and well- characterized Ni-MWCNT. 2. Determine the importance of Ni-MWCNT surface properties (amount of Nickel and surface charge) to cause cytotoxicity and degree of activation of the NLRP3 inflammasome using THP-1 cells. Finally, we will establish that internalization of MWCNT-Ni followed by lysosomal membrane disruption and Cathepsin B release is the initiating event in activation of the NLRP3 inflammasome. Thus, information from this study will be important in determining characteristics of safe ENM and establish mechanisms of action and may lead to an in vitro screening platform using THP-1 cells.