Bioactivity and mechanistic studies using a comprehensive and well characterized

描述(由申请人提供)：许多碳纳米管已被证明可引起肺部炎症和纤维化，但导致这些影响的机制尚不清楚。此外，制造多种不同形式的碳纳米管(CNT)的能力使人们迫切需要更好地了解碳纳米管的安全性和危险等级。我们已经描述了CNT的激活引起吞噬溶酶体膜通透性(LMP)，导致组织蛋白酶B的释放和NLRP3炎性小体的激活。然而，导致LMP和其他控制炎症反应的事件(例如自噬)的机制还没有得到很好的描述，因此限制了纳米毒理学领域的进展。该项目的总体目标是开发具有良好特性的库 多壁和单壁碳纳米管具有可控的性质(物理和化学)，代表了最具商业可行性的形式，并使用这些材料来洞察碳纳米管在体外和体内的毒性机制和结构活性关系。我们的中心假设是，生物反应将依赖于碳纳米管的特定性质，这些性质调节吞噬小体的膜通透性(LMP)和自噬。此外，我们认为碳纳米管通过影响胆固醇的转运而引起LMP。因此，我们认为，基于这些特性，我们将能够预测CNT的促炎和促纤维化活性。中心假设将通过以下目标进行检验：目标1：建立具有特定物理化学特性的完全表征的CNT的综合文库。目的2：确定目标1中开发的CNT的生物活性机制，并表征CNT对LMP、NLRP3的摄取、相对生物活性(IL-1？释放)和自噬。目的3：探讨选择性CNT的体内病理机制。我们跨学科团队的长期目标是开发一种基于碳纳米管的物理化学性质进行毒性预测的方法。这一策略随后可用于危险等级以及具有高商业潜力的碳纳米管形式的安全设计。

DESCRIPTION (provided by applicant): Many carbon nanotubes have been shown to cause lung inflammation and fibrosis, but the mechanisms responsible for these effects are not well understood. Furthermore, the ability to manufacture carbon nanotubes (CNT) in many different formats has created an urgency to develop better understanding of CNT safety and hazard ranking. We have described that CNT activated cause phagolysosomal membrane permeability (LMP) leading to release of cathepsin B and NLRP3 inflammasome activation. However, the mechanisms responsible for causing LMP and additional events controlling the inflammatory response (e.g., autophagy) are not well described, thus limiting progress in the field of nanotoxicology. The overall objective of this project is to develop well-characterized libraries of multi-walled and single-walled CNT with controlled properties (physical and chemical) that represent the most commercially viable forms and use these materials to provide insight into a mechanistic understanding and structure activity relationship of CNT toxicity both in vitro and in vivo. Our central hypothesis is that the biological responses will be dependent on specific properties of CNT and these properties regulate phagolysosomal membrane permeability (LMP) and autophagy. Furthermore, we propose that CNT cause LMP by affecting cholesterol trafficking. Therefore, we propose that we will be able to predict the proinflammatory and profibrotic activity of CNT based on these properties. The central hypothesis will be tested with the following aims: Aim 1: Develop a comprehensive library of fully characterized CNT with specific physicochemical characteristics. Aim 2: Determine the mechanism of bioactivity of CNT developed in Aim 1 and characterize the uptake, relative bioactivity of the CNT to cause LMP, NLRP3 activation (IL-1? release) and autophagy. Aim 3: Evaluate the mechanism of in vivo pathology of selected CNT. The long-term goal of our interdisciplinary team is to develop an approach to toxicity prediction based on physicochemical properties of CNT. This strategy can then be used for hazard ranking as well as safe design of the CNT forms with high commercial potential.