Evaluating polyethyleneimine/DNA nanoparticles-mediated damage to cellular organelles using endoplasmic reticulum stress profile

Abstract

Gene therapy has emerged as an influential tool for treating the genetic and specific acquired disorders. Among all kinds of gene delivery systems, the cationic polymer polyethyleneimine (PEI) is considered as a promising non-viral gene delivery vector, although there are still concerns about its magnitude of cytotoxicity. While any cell insult leads to unfolded/misfolded protein accumulation and its consequent unfold protein response, evaluating the expression profile of ER-stress genes would be a sensitive indicator of cell stress. Beside cytotoxicity assays, real-time RT-PCR was used to investigate the effects of PEI nanoparticles on the endoplasmic reticulum. Treating Neuro2A cells revealed that PEI can induce cell toxicity in a concentration-dependent manner. Also, It increased the transcript levels of Grp78 (Bip), Atf4 and Chop, and splicing of Xbp1. To further optimize the transfection properties in Neuro2A cells, PEI was used to deliver a plasmid DNA containing GFP reporter. While different PEI/plasmid ratios revealed similar transfection efficiency, increasing the PEI/plasmid ratio led to induction of ER-stress markers. These results underscored that beside the effectiveness of PEI, using the lowest possible ratio of PEI/plasmid would minimize the detrimental effects of PEI on cells and confer it a beneficial therapeutic importance in nucleic acid delivery.

The unfolded protein response to PI*Z alpha‐1 antitrypsin in human hepatocellular and murine models

“…Transfection reagents, both cationic lipids and polycationic polymers, can cause ER stress. [ 28 , 29 , 30 ] Transduced genes can be overexpressed depending on the copy number and the promoters. An overexpressed protein, even a normal protein, can aggregate together in the ER and harm the host cells.…”

Decoding cold ischaemia time impact on kidney graft: the kinetics of the unfolded protein response pathways

“…In conclusion, our results demonstrate that UPR could be a critical pathway underlying the relationship between CIT and graft outcome. UPR profiling indeed appears to be a critical tool to evaluate the level of lesion within the cell [52]. Our data highlight the potential for UPR-based strategies in transplantation: (a) on the one hand, measuring the activation status of each branch will provide important information on the lesion advance, and the ability of the organ to handle the injury, hence allowing anticipation of outcome; (b) on the other, therapies based on modulating each branch will be able to moderate the activation of negative pathways (ATF6, XBP1s) while promoting beneficial pathways (ATF4), hence reducing lesions and improving recovery.…”

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