Investigation into Mechanisms NLRP3 Activation by Nickel Associated Multi-Walled

镍伴生多壁激活 NLRP3 的机制研究

• 批准号: 8522283
• 负责人: Teri Alyn Girtsman
• 金额: $ 3.62万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-15 至 2014-04-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8522283
• 关键词: Alveolar Macrophages; Animal Model; Animals; Area; Benign; Biological; Biological Assay; Biological Testing; Carbon Nanotubes; Cathepsins B; Cell Line; Cells; Characteristics; Charge; Cosmetics; Data; Development; Drug Delivery Systems; Engineering; Evaluation; Event; Exposure to; Future; Goals; Growth; Health; Home environment; Human; Hybrids; In Vitro; Individual; Inflammation; Inflammatory; Investigation; Ions; Knowledge; Laboratories; Lead; Lung; Lung Inflammation; Lysosomes; Medical; Membrane; Metals; Modeling; Mus; Nickel; Outcome; Pathology; Phagocytosis; Population; Production; Property; Pulmonary Pathology; Risk; Signal Transduction; Surface; Surface Properties; System; Testing; Toxic effect; Toxicity Tests; Variant; Work; base; catalyst; chemical property; cost; cytokine; cytotoxicity; design; high throughput screening; in vivo; man; metal oxide; nanomaterials; nanoparticle; particle; physical property; predictive modeling; screening; tool

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.

描述（由申请人提供）：某些工程纳米材料（ENM）已被证明会在动物模型中引起显着的肺部病理变化，引起人们对随着使用和接触的增加而出现的人类健康影响的担忧。然而，尚未建立基于 ENM 物理和表面特性的机械预测模型来帮助保护人类健康。我们实验室的其他工作表明，体外 NLRP3 炎症小体 (NLRP3) 激活与体内炎症之间存在关联。在初步数据中，我们已经表明，镍相关多壁碳纳米管（Ni-MWCNT）在体外使用原代肺泡巨噬细胞（AM）或THP-1细胞激活NLRP3的能力与肺部炎症和病理学之间可能存在相关性。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 的体内炎症潜力与溶酶体破坏活性和 NLRP3 激活密切相关。根据初步数据，我们认为多壁碳纳米管的炎症潜力将取决于相关镍的含量。因此，我们的目标是： 1. 确定 AM 中的 NLRP3 炎性体活性与小鼠暴露于明确且明确表征的 Ni-MWCNT 后的肺部病理学之间的关系。 2. 使用 THP-1 细胞确定 Ni-MWCNT 表面特性（镍量和表面电荷）对引起细胞毒性和 NLRP3 炎性体激活程度的重要性。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 释放是 NLRP3 炎性体激活的起始事件。因此，本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。