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会导致肺部炎症和纤维化，但对这些影响的机制还不清楚。此外，以许多不同形式制造碳纳米管（CNT）的能力已经产生了更好地理解CNT安全性和危害等级的紧迫性。我们已经描述了CNT活化引起吞噬溶酶体膜通透性（LMP），导致组织蛋白酶B的释放和NLRP 3炎性体活化。然而，负责引起LMP和控制炎症反应的其他事件（例如，自噬）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特色鲜明的图书馆， 具有受控性质（物理和化学）的多壁和单壁CNT，代表最具商业可行性的形式，并使用这些材料来深入了解CNT体外和体内毒性的机制和结构活性关系。我们的中心假设是，生物反应将取决于CNT的特定性质，这些性质调节吞噬溶酶体膜通透性（LMP）和自噬。此外，我们认为CNT通过影响胆固醇运输引起LMP。因此，我们建议，我们将能够预测基于这些属性的CNT的促炎和促纤维化活性。中心假设将测试以下目标：目标1：开发一个全面的图书馆，充分表征碳纳米管与特定的物理化学特性。目标二：确定目标1中开发的CNT的生物活性机制，并表征CNT引起LMP、NLRP 3激活（IL-1？释放）和自噬。目的3：评价所选CNT的体内病理学机制。我们跨学科团队的长期目标是开发一种基于CNT理化性质的毒性预测方法。然后，该策略可用于危险等级以及具有高商业潜力的CNT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.