Cellular entry of nanoparticles via serum sensitive clathrin-mediated endocytosis, and plasma membrane permeabilization

Rappoport, Joshua Z.; Smith, Derek; Giroud,; Wiggins,; Mazzolin,; Dyson,

doi:10.2147/ijn.s29334

Cited by 40 publications

(24 citation statements)

References 31 publications

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“…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).…”

Section: Discussionsupporting

confidence: 93%

“…The NPs are known to interact with molecules at the cell surface, and can subsequently be internalized specifically through membrane interactions, via receptor-ligand mediated active processes or electrostatic interactions, or through nonspecific routes such as passive fluid encapsulation within vesicles during internalization processes. A series of calculations were performed to determine the potential for uptake through direct membrane interaction versus uptake due to passive incorporation into vesicles in the liquid phase (Smith et al 2012).…”

Section: Mechanism Of Uptake: Membrane-bound or Fluid Phase?mentioning

confidence: 99%

“…The mean inhibition of each pathway was >70% in each cell line and with each siRNA as determined from the expression levels of the knocked-down proteins. Cav1 expression in macrophages has been debated, and some macrophages, Smith et al (2012). (A) The equations can be substituted to determine the most likely means of NP cellular entry: fluid phase, or receptor mediated.…”

Section: Mechanism Of Spion Uptake Into Different Cancer Cell Lines: mentioning

confidence: 99%

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Mechanisms for cellular uptake of nanosized clinical MRI contrast agents

2020

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Engineered Nanomaterials (NMs), such as Superparamagnetic Iron Oxide Nanoparticles (SPIONs), offer significant benefits in a wide range of applications, including cancer diagnostic and therapeutic strategies. However, the use of NMs in biomedicine raises safety concerns due to lack of knowledge on possible biological interactions and effects. The initial basis for using SPIONs as biomedical MRI contrast enhancement agents was the idea that they are selectively taken up by macrophage cells, and not by the surrounding cancer cells. To investigate this claim, we analyzed the uptake of SPIONs into well-established cancer cell models and benchmarked this against a common macrophage cell model. In combination with fluorescent labeling of compartments and siRNA silencing of various proteins involved in common endocytic pathways, the mechanisms of internalization of SPIONs in these cell types has been ascertained utilizing reflectance confocal microscopy. Caveolar mediated endocytosis and macropinocytosis are both implicated in SPION uptake into cancer cells, whereas in macrophage cells, a clathrin-dependant route appears to predominate. Colocalization studies confirmed the eventual fate of SPIONs as accumulation in the degradative lysosomes. Dissolution of the SPIONs within the lysosomal environment has also been determined, allowing a fuller understanding of the cellular interactions, uptake, trafficking and effects of SPIONs within a variety of cancer cells and macrophages. Overall, the behavior of SPIONS in non-phagocytotic cell lines is broadly similar to that in the specialist macrophage cells, although some differences in the uptake patterns are apparent. ARTICLE HISTORY

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Section: Discussionsupporting

confidence: 93%

Section: Mechanism Of Uptake: Membrane-bound or Fluid Phase?mentioning

confidence: 99%

Section: Mechanism Of Spion Uptake Into Different Cancer Cell Lines: mentioning

confidence: 99%

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Mechanisms for cellular uptake of nanosized clinical MRI contrast agents

2020

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“…Although recent studies have demonstrated that the physicochemical properties of a nanocarrier modulates its intracellular trafficking, 55,56 it still remained unclear, prior to our current study, as to how the presence of multiple copies of a targeting ligand on the surface of a nanocarrier, i.e. its multivalency, influences its intracellular trafficking and subcellular accumulation, relative to its non-targeted counterpart, or even relative to the targeting ligand alone.…”

Section: Resultsmentioning

confidence: 96%

Modulation of hydrogel nanoparticle intracellular trafficking by multivalent surface engineering with tumor targeting peptide

et al. 2013

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Surface engineering of a hydrogel nanoparticle (NP) with the tumor-targeting ligand, F3 peptide, enhances both the NP’s binding affinity for, and internalization by, nucleolin overexpressing tumor cells. Remarkably, the F3-functionalized NPs consistently exhibited significantly lower trafficking to the degradative lysosomes than the non-functionalized NPs, in the tumor cells, after internalization. This is attributed to the non-functionalized NPs, but not the F3-functionalized NPs, being co-internalized with Lysosome-associated Membrane Protein-1 (LAMP1) from the surface of the tumor cells. Furthermore, it is shown that the intracellular trafficking of the F3-functionalized NPs differs significantly from that of the single F3 peptides. This has important implications for designing effective, chemically-responsive, controlled-release and multifunctional nanodrugs for multi-drug-resistant cancers.

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“…Immediately before transfection, media were aspirated and replaced with 100 μl Opti-MEM + 2% FBS. The serum concentration was 2% FBS for the transfection experiments because low serum conditions increases cell uptake and minimizes the introduction of experimental variability due to rapid cell proliferation 24 . Fresh or reconstituted lyophilized polyplexes containing 150 ng pDNA were then added to each well.…”

Section: Methodsmentioning

confidence: 99%

Enhanced performance of plasmid DNA polyplexes stabilized by a combination of core hydrophobicity and surface PEGylation

et al. 2014

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Nonviral gene therapy has high potential for safely promoting tissue restoration and for treating various genetic diseases. One current limitation is that conventional transfection reagents such as polyethylenimine (PEI) form electrostatically stabilized plasmid DNA (pDNA) polyplexes with poor colloidal stability. In this study, a library of poly(ethylene glycol-b-(dimethylaminoethyl methacrylate-co-butyl methacrylate)) [poly(EG-b-(DMAEMA-co-BMA))] polymers were synthesized and screened for improved colloidal stability and nucleic acid transfection following lyophilization. When added to pDNA in the appropriate pH buffer, the DMAEMA moieties initiate formation of electrostatic polyplexes that are internally stabilized by hydrophobic interactions of the core BMA blocks and sterically stabilized against aggregation by a PEG corona. The BMA content was varied from 0% to 60% in the second polymer block in order to optimally tune the balance of electrostatic and hydrophobic interactions in the polyplex core, and polymers with 40 and 50 mol% BMA achieved the highest transfection efficiency. Diblock copolymers were more stable than PEI in physiologic buffers. Consequently, diblock copolymer polyplexes aggregated more slowly and followed a reaction-limited colloidal aggregation model, while fast aggregation of PEI polyplexes was governed by a diffusion-limited model. Polymers with 40% BMA did not aggregate significantly after lyophilization and produced up to 20-fold higher transfection efficiency than PEI polyplexes both before and after lyophilization. Furthermore, poly(EG-b-(DMAEMA-co-BMA)) polyplexes exhibited pH-dependent membrane disruption in a red blood cell hemolysis assay and endosomal escape as observed by confocal microscopy.Lyophilized polyplexes made with the lead candidate diblock copolymer (40% BMA) also successfully transfected cells in vitro following incorporation into gas-foamed polymeric scaffolds. In summary, the enhanced colloidal stability, endosomal escape, and resultant high transfection efficiency of poly(EG-b-(DMAEMA-co-BMA))-pDNA polyplexes underscores their potential utility both for local delivery from scaffolds as well as systemic, intravenous delivery.

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Cellular entry of nanoparticles via serum sensitive clathrin-mediated endocytosis, and plasma membrane permeabilization

Cited by 40 publications

References 31 publications

Mechanisms for cellular uptake of nanosized clinical MRI contrast agents

Mechanisms for cellular uptake of nanosized clinical MRI contrast agents

Modulation of hydrogel nanoparticle intracellular trafficking by multivalent surface engineering with tumor targeting peptide

Enhanced performance of plasmid DNA polyplexes stabilized by a combination of core hydrophobicity and surface PEGylation

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