Protein Corona Modulates Uptake and Toxicity of Nanoceria via Clathrin-Mediated Endocytosis

Abstract

Particles present in diesel exhaust have been proposed as a significant contributor to the development of acute and chronic lung diseases, including respiratory infection and allergic asthma. Nanoceria (CeO2 nanoparticles) are used to increase fuel efficiency in internal combustion engines, are present in exhaust fumes, and could affect cells of the airway. Components from the environment such as biologically derived proteins, carbohydrates, and lipids can form a dynamic layer, commonly referred to as the "protein corona" which alters cellular nanoparticle interactions and internalization. Using confocal reflectance microscopy, we quantified nanoceria uptake by lung-derived cells in the presence and absence of a serum-derived protein corona. Employing mass spectrometry, we identified components of the protein corona, and demonstrated that the interaction between transferrin in the protein corona and the transferrin receptor is involved in mediating the cellular entry of nanoceria via clathrin-mediated endocytosis. Furthermore, under these conditions nanoceria does not affect cell growth, viability, or metabolism, even at high concentration. Alternatively, despite the antioxidant capacity of nanoceria, in serum-free conditions these nanoparticles induce plasma membrane disruption and cause changes in cellular metabolism. Thus, our results identify a specific receptor-mediated mechanism for nanoceria entry, and provide significant insight into the potential for nanoparticle-dependent toxicity.

Mechanisms for cellular uptake of nanosized clinical MRI contrast agents

“…This is in accordance with other results investigating the time and concentration dependence of uptake for a variety of NPs (e.g. polystyrene and gold) and cell lines (Johnston et al 2010;Trono et al 2011;Smith et al 2012;Mazzolini et al 2015).…”

“…Tf is known to bind to many different NPs and thus may offer a mechanism of entry for corona coated SPIONs in these cell types (Raynal et al 2004;Arbab et al 2005). Corona studies performed on the SPIONs used here identified the presence of Tf, indicating this as a potential mechanism of entry into the THP-1 cells through the CME pathway (Supplementary Figure S8); likewise the presence of Tf bound to CeO 2 NPs has been shown to be involved in receptor mediated endocytosis via CME (Mazzolini et al 2015).…”

“…Reflectance intensity was measured through automatic background subtraction and manual delineation due to the high confluence of cells at these longer time points. A decrease in reflectance intensity was observed during the 96 hours in A459 cells (Figure 7(A)) but this may be due to NP distribution to daughter cells during mitosis (Mazzolini et al 2015). During the dissolution study in ALF, decreases in the observed diameter were observed from T ¼ 0 (78.1 ± 3.3 nm) to T ¼ 96 (30.5 ± 4.4 nm) indicating dissolution of the NPs in ALF (Figure 7(B)).…”

Size, Shape, and Protein Corona Determine Cellular Uptake and Removal Mechanisms of Gold Nanoparticles

“…Jiang et al proved that zwitterionic quantum dot nanoparticles enter mainly via the clathrin‐mediated endocytosis. Mazzolini et al results identify a specific receptor‐mediated mechanism of CeO 2 nanoparticles. After the NPs enter the cells, they go through early endosome and late endosomes, locate in the lysosomes, and finally excrete from cells by lysosomes.…”

Valproic Acid Induces Endocytosis-Mediated Doxorubicin Internalization and Shows Synergistic Cytotoxic Effects in Hepatocellular Carcinoma Cells

“…Endocytic trafficking involves a series of steps including endocytosis, cargo sorting and processing, intracellular membrane fusion and fission, vesicle mobility, and exocytosis [35]; the endocytic pathways include clathrin-dependent endocytosis, caveolae-dependent endocytosis, macropinocytosis, and phagocytosis. Recently, the pathways of clathrin and caveolae receptor-mediated endocytosis were well studied in cellular membrane trafficking for cancer therapy [36,37,38,39]. …”