Different Sensitivity of Macrophages to Phospholipidosis Induction by Amphiphilic Cationic Drugs

Abstract

Phospholipidosis (PLD), the intracellular accumulation of phospholipids, is an adaptive response to toxic stimuli and serves as an important parameter in the biological assessment of compounds. Cationic amphiphilic drugs are the main inducers of PLD and may impair the function of alveolar macrophages. In vivo and in vitro models are used for PLD screening but the choice of the cellular model may be important because PLD develops in a cell- and species-specific manner. In this study, a panel of different staining (LysoSensor, Acridine Orange, Nile Red, HCS LipidTOX, LysoID) was evaluated in murine (DMBM-2, J774, RAW264.7) and human (THP-1, monocyte-derived macrophages from peripheral blood) cells to identify the most sensitive and easy to analyze staining method and to detect species-specific differences in the reaction pattern. Amiodarone and chloroquine served as inducers of PLD. High content screening was used to compare number, area, and intensity of the staining. Due to the fast staining protocol and the sensitivity of the detection, LysoID proved to be the most suitable dye of the testing. The lower induction of PLD by chloroquine reported in vivo was also seen in this study. THP-1 macrophages, followed by DMBM-2 cells, produced the most similar reaction pattern to human monocyte-derived macrophages.

Integrating distribution kinetics and toxicodynamics to assess repeat dose neurotoxicity in vitro using human BrainSpheres: a case study on amiodarone

“…In particular, the upregulation of the genes MSMO1 , IDI1 , LSS , and HMGCR suggested enhanced synthesis of cholesterol, and the upregulation of FASN suggested an increased synthesis of fatty acids that may in turn induce phospholipidosis ( Sawada et al, 2005 ; Antherieu et al, 2011 ). This would be in agreement with the reported induction of phospholipidosis by AMI, in various cell types, such as macrophages, alveolar epithelial cells, and hepatocytes ( Lewis et al, 1990 ; Nonoyama and Fukuda, 2008 ; Kapatou et al, 2010 ; Ohlinger et al, 2020 ). However, this upregulation of lipogenic genes was not accompanied by the upregulation of PLIN s that are essential for lipid droplet building and, therefore, lipid storage.…”

Vitamin D3and carbamazepine protect againstClostridioides difficileinfection in mice by restoring macrophage lysosome acidification

“…It is also notable that, in this study, human monocyte-derived macrophages (THP-1) and murine macrophages (RAW 264.7) reacted similarly to TcdB in terms of SQSTM1/p62 accumulation and cytokine response but exhibited different sensitivities. A recent study also showed that human and murine macrophages showed different sensitivities to lysosomotropic agents [ 45 ]. Another important aspect that warrants further investigation is whether TcdB-induced lysosome dysfunction during CDI could instigate other downstream pro-inflammatory pathways, including the AIM2 (absent in melanoma 2)/NLRP3 (NLR family pyrin domain containing 3)-inflammasomes, the CGAS (cyclic GMP-AMP synthase)-STING1 (stimulator of interferon response cGAMP interactor 1) pathway and mitochondrial homeostasis, all of which are important for linking lysosome dysfunction to cytokine response [ 23 ].…”

Cholesterol nanoarchaeosomes for alendronate targeted delivery as an anti-endothelial dysfunction agent

“…In contrast, J774A.1 cells express a phenotype close to M0 with greater expression of SRAI/II [ 41 ]. Therefore, the effect of nanoarchaeosomes may differ depending on, for instance, the cell genotype because of differences in phospholipid processing [ 45 ] and on the various factors that modulate the expression of SRAI/II. We observed here that nanoARC(ALN) was much less cytotoxic for THP-1 macrophages than for J774A1 cells, probably, because the expression of SRA1 in THP-1 macrophages (and subsequent internalization) is reported to be lower than in J774A1 macrophages [ 43 ].…”