Health effects of manufactured nanoparticles (NP): molecular and cellular biology and toxicology (NanoBioTox)

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This project aims to advance our understanding of the interactions of nanoparticles (NPs) with proteins, tissue and cells of the respiratory tract. In the previous funding period, we have developed a toolkit of NPs with well-defined, specific properties. We have analyzed the binding of model proteins to our NPs with respect to kinetics and equilibrium. With in-vitro studies, we have demonstrated that NP surface properties govern their uptake by and their final destination inside cells, and also cytotoxic effects. We further provided evidence that, in vivo, NP surface properties, including protein adsorption, likewise affect their distribution within the body. Based on these results, we plan to continue our studies by including experiments aimed at elucidating the fate of NPs after their uptake by cells, specifically, with regard to the “protein corona” enshrouding the NPs. Our work is structured in three major areas. The first one ("NP toolbox") aims at the synthesis and characterization of water-soluble functional NPs. Modification of the protein corona around NPs will play a major role. The second topic ("NP-protein complexes") focuses on NP interactions with proteins and other biomolecules of body fluids. Special attention will be devoted to the epithelial lining fluid, an aqueous liquid layer containing glycoproteins and pulmonary surfactant components, and to the continuous pulmonary surfactant film at the air-liquid interface of the internal lung surface. We will quantify kinetic and equilibrium behavior of key proteins binding to NPs as a function of the specific physical and chemical properties of our NPs. Moreover, we will examine the structure of the protein corona in detail. Do proteins adsorb to NPs in well defined orientations or randomly? Is the protein structure conserved upon adsorption, or do adsorbed proteins develop toward denatured conformations? In the third area ("NP-cell interactions and cellular responses") we study the interaction of NPs with cells from lung tissue ex vivo, lung cells in vitro and their organelles. Also here, the influence of the protein corona will be investigated. Will the structure of the protein corona be preserved after NP uptake by the cell? Finally, we will investigate if and how adsorbed proteins affect toxic effects elicited by NPs.