Protective role of autophagy in branched polyethylenimine (25K)- and poly(L-lysine) (30–70K)-induced cell death

Lin, Chia Wei; Jan, Ming‐Shiou; Kuo, Jung hua Steven; Hsu, Li Jin; Lin, Yee Shin

doi:10.1016/j.ejps.2012.09.007

Cited by 36 publications

(45 citation statements)

References 29 publications

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“…Typical cellular responses to mitochondrial damage are autophagy/mitophagy, apoptosis, and necrosis. Apoptosis and autophagy are mutually inhibitory, with apoptosis leading to cell death, while autophagy possibly serves a protective role in PEI cytotoxicity …”

Section: Discussionsupporting

confidence: 74%

“…Apoptosis and autophagy are mutually inhibitory, 45 with apoptosis leading to cell death, while autophagy possibly serves a protective role in PEI cytotoxicity. 46 Altogether, literature review of grouped screen hit compounds suggests that many priming effects may be due to modulation of the cellular oxidative stress response to PEI transfection, in particular mitochondrial dysfunction. It is plausible that certain antioxidants like resveratrol 29,30 rescue the cell from PEI-induced mitochondrial dysfunction through modulation of autophagy, improving transfection, while certain antibiotics that reduce transfection, such as cephalosporins, 47 may be inhibiting transfection by increasing mitochondrial dysfunction and oxidative stress.…”

Section: Mitochondrial Dysfunction Oxidative Stress and Autophagymentioning

confidence: 99%

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High‐throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

Nguyen

Beyersdorf

Riethoven

et al. 2016

Bioengineering & Transla Med

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Nonviral gene delivery methods are advantageous over viral vectors in terms of safety, cost, and flexibility in design and application, but suffer from lower gene transfer efficiency. In addition to modifications to nucleic acid design and nonviral carriers, new tools are sought to enhance transfection. Priming is the pharmacological modulation of transfection efficiency and transgene expression, and has demonstrated transfection increase in several compounds, for example, chloroquine and glucocorticoids. To develop a library of transfection priming compounds, a high‐throughput screen was performed of the NIH Clinical Collection (NCC) to identify clinical compounds that prime polyethylenimine (PEI) transfection. HEK293T cells were treated with priming compounds, then transfected with enhanced green fluorescent protein (EGFP)‐encoding plasmid by PEI. After 48‐hr culture, primed and transfected cells were assayed for transfection, cell proliferation, and cell viability by fluorescence measurement of EGFP reporter, Hoechst 33342 nuclei stain, and resazurin metabolic assay. From the microscope image analysis and microplate measurements, transfection fold‐changes were determined, and compounds resulting in statistically significant transfection fold‐change were identified. NCC compounds were clustered using PubChem fingerprint similarity by Tanimoto coefficients in ChemmineTools. Fold‐changes for each compound were linked to drug clusters, from which drug classes that prime transfection were identified. Among the identified drugs classes that primed transfection increases were antioxidants, GABAA receptor modulators, and glucocorticoids. Resveratrol and piceid, stilbenoid antioxidants found in grapes, and zolpidem, a GABAA modulator, increased transfection nearly three‐fold. Literature indicate interaction of the identified transfection priming drug clusters with mitochondria, which may modulate mitochondrial dysfunction known to be associated with PEI transfection.

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Section: Discussionsupporting

confidence: 74%

Section: Mitochondrial Dysfunction Oxidative Stress and Autophagymentioning

confidence: 99%

High‐throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

Nguyen

Beyersdorf

Riethoven

et al. 2016

Bioengineering & Transla Med

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“…Numerous types of nanomaterials, including PEI and PAMAM, have been previously shown to induce autophagy in various cell lines [21–24]. Though the exact role of this cellular response has not been fully characterized, many studies have postulated nanomaterial-induced autophagy to function as a means of eliminating or extruding toxic materials from the cell, and as a last-ditch mechanism for cell survival before the cell undergoes apoptosis or necrosis [25].…”

Section: Discussionmentioning

confidence: 99%

A mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell lines

et al. 2017

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BackgroundGold–polyamidoamine (AuPAMAM) has previously been shown to successfully transfect cells with high efficiency. However, we have observed that certain cell types are more amenable to Au–PAMAM transfection than others. Here we utilized two representative cell lines—a “difficult to transfect” CT26 cell line and an “easy to transfect” SK-BR3 cell line—and attempted to determine the underlying mechanism for differential transfection in both cell types. Using a commonly established poly-cationic polymer similar to PAMAM (polyethyleneimine, or PEI), we additionally sought to quantify the relative transfection efficiencies of each vector in CT26 and SK-BR3 cells, in the hopes of elucidating any mechanistic differences that may exist between the two transfection vectors.ResultsA comparative time course analysis of green fluorescent protein reporter-gene expression and DNA uptake was conducted to quantitatively compare PEI- and AuPAMAM-mediated transfection in CT26 and SK-BR3, while flow cytometry and confocal microscopy were used to determine the contribution of cellular uptake, endosomal escape, and cytoplasmic transport to the overall gene delivery process. Results from the time course analysis and flow cytometry studies revealed that initial complex uptake and cytoplasmic trafficking to the nucleus are likely the two main factors limiting CT26 transfectability.ConclusionsThe cell type-dependent uptake and intracellular transport mechanisms impacting gene therapy remain largely unexplored and present a major hurdle in the application-specific design and efficiency of gene delivery vectors. This systematic investigation offers insights into the intracellular mechanistic processes that may account for cell-to-cell differences, as well as vector-to-vector differences, in gene transfectability.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-017-0271-8) contains supplementary material, which is available to authorized users.

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“…PEI-induced autophagy plays a protective role in cell survival. 14,15 However, effects of autophagy on PEI degradation remain unknown.…”

Section: Introductionmentioning

confidence: 99%

Autophagy promotes degradation of polyethyleneimine–alginate nanoparticles in endothelial progenitor cells

Wang

Tan

Wang

et al. 2017

IJN

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Polyethyleneimine (PEI)–alginate (Alg) nanoparticle (NP) is a safe and effective vector for delivery of siRNA or DNA. Recent studies suggest that autophagy is related to cytotoxicity of PEI NPs. However, contribution of autophagy to degradation of PEI–Alg NPs remains unknown. CD34 + VEGFR-3 + endothelial progenitor cells isolated from rat bone marrow were treated with 25 kDa branched PEI modified by Alg. After treatment with the NPs, morphological changes and distribution of the NPs in the cells were examined with scanning and transmission electron microscopies. Cytotoxicity of the NPs was analyzed by reactive oxygen species (ROS) production, lactate dehydrogenase leakage and induction of apoptosis. The level of autophagy was assessed with expression of Beclin-1 and LC3 and formation of autophagic structures and amphisomes. Colocalization of LC3-positive puncta and the NPs was determined by LC3–GFP tracing. Cytotoxicity of PEI NPs was reduced greatly after modification with Alg. PEI–Alg NPs were distributed in mitochondria, rough endoplasmic reticula and nuclei as well as cytoplasm. After phagocytosis of the NPs, expression of Beclin-1 mRNA and LC3 protein was upregulated, and the number of LC3-positive puncta, autophagic structures and amphisomes increased significantly. The number of lysosomes also increased obviously. There were LC3-positive puncta in nuclei, and some puncta were colocalized with the NPs. These results demonstrate that the activated autophagy promotes degradation of PEI–Alg NPs via multiple pathways.

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Protective role of autophagy in branched polyethylenimine (25K)- and poly(L-lysine) (30–70K)-induced cell death

Cited by 36 publications

References 29 publications

High‐throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

High‐throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

A mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell lines

Autophagy promotes degradation of polyethyleneimine–alginate nanoparticles in endothelial progenitor cells

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Protective role of autophagy in branched polyethylenimine (25K)- and poly(L-lysine) (30–70K)-induced cell death

Cited by 36 publications

References 29 publications

High‐throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

High‐throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

A mechanistic investigation exploring the differential transfection efficiencies between the easy-to-transfect SK-BR3 and difficult-to-transfect CT26 cell lines

Autophagy promotes degradation of polyethyleneimine&ndash;alginate nanoparticles in endothelial progenitor cells

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Autophagy promotes degradation of polyethyleneimine–alginate nanoparticles in endothelial progenitor cells