Intestinal Transcytosis of a Protein Cargo and Nanoparticles Mediated by a Non-Toxic Form of Pseudomonas aeruginosa Exotoxin A

Li, Ruiying; Laurent, Floriane; Taverner, Alistair; Mackay, Julia D.; Bank, Paul De; Mrsny, Randall J.

doi:10.3390/pharmaceutics13081171

Cited by 6 publications

(4 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, molecules larger than 700 Da would enter the cell via active transport [9]. The transcellular pathway (a type of active transport) involves the endocytic uptake at the apical site, intracellular vesicle traffic, and basolateral efflux of the content [12]. This phenomenon occurs both in the enterocytes and in the M cells of Peyer patches localized in the intestinal epithelium [13].…”

Section: Introductionmentioning

confidence: 99%

Zein-Based Nanoparticles as Oral Carriers for Insulin Delivery

et al. 2021

View full text Add to dashboard Cite

Zein, the major storage protein from corn, has a GRAS (Generally Regarded as Safe) status and may be easily transformed into nanoparticles, offering significant payloads for protein materials without affecting their stability. In this work, the capability of bare zein nanoparticles (mucoadhesive) and nanoparticles coated with poly(ethylene glycol) (mucus-permeating) was evaluated as oral carriers of insulin (I-NP and I-NP-PEG, respectively). Both nanocarriers displayed sizes of around 270 nm, insulin payloads close to 80 µg/mg and did not induce cytotoxic effects in Caco-2 and HT29-MTX cell lines. In Caenorhabditis elegans, where insulin decreases fat storage, I-NP-PEG induced a higher reduction in the fat content than I-NP and slightly lower than the control (Orlistat). In diabetic rats, nanoparticles induced a potent hypoglycemic effect and achieved an oral bioavailability of 4.2% for I-NP and 10.2% for I-NP-PEG. This superior effect observed for I-NP-PEG would be related to their capability to diffuse through the mucus layer and reach the surface of enterocytes (where insulin would be released), whereas the mucoadhesive I-NP would remain trapped in the mucus, far away from the absorptive epithelium. In summary, PEG-coated zein nanoparticles may be an interesting device for the effective delivery of proteins through the oral route.

show abstract

Section: Introductionmentioning

confidence: 99%

Zein-Based Nanoparticles as Oral Carriers for Insulin Delivery

et al. 2021

View full text Add to dashboard Cite

show abstract

“…It is the reason behind the increased mortality rates among experimentally infected animals where a single injection of 80 ng was sufficient to induce severe liver necrosis and swelling, hemorrhage in the lungs and kidneys within 48 h of exposure (Wretlind and Pavlovskis 1981 ). A recent study proved the potential applications of nontoxic P. aeruginosa exotoxin A to facilitate the oral delivery of macromolecular therapeutics under conditions of covalent or non-covalent association (Li et al 2021 ). Recently, Shadman et al have developed monoclonal antibodies against exotoxin A as an alternative approach to control infection by this life threatening pathogen (Shadman et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

ADP-ribosyl transferase activity and gamma radiation cytotoxicity of Pseudomonas aeruginosa exotoxin A

et al. 2021

View full text Add to dashboard Cite

This work explores the ADP-ribosyltransferase activity of Pseudomonas (P.) aeruginosa exotoxin A using the guanyl hydrazone derivative, nitrobenzylidine aminoguanidine (NBAG) and the impact of gamma radiation on its efficacy. Unlike the conventional detection methods, NBAG was used as the acceptor of ADP ribose moiety instead of wheat germ extract elongation factor 2. Exotoxin A was extracted from P. aeruginosa clinical isolates and screened for toxA gene using standard PCR. NBAG was synthesized using aminoguanidine bicarbonate and 4-nitrobenzaldehyde and its identity has been confirmed by UV, FTIR, Mass and 13C-NMR spectroscopy. The ADP-ribosyl transferase activity of exotoxin A on NBAG in the presence of Nicotinamide adenine dinucleotide (NAD+) was recorded using UV spectroscopy and HPLC. In vitro ADP-ribosyl transferase activity of exotoxin A protein extract was also explored by monitoring its cytotoxicity on Hep-2 cells using sulforhodamine B cytotoxicity assay. Bacterial broths were irradiated at 5, 10, 15, 24 Gy and exotoxin A protein extract activity were assessed post exposure. Exotoxin A extract exerted an ADP-ribosyltransferase ability which was depicted by the appearance of a new ʎmax after the addition of exotoxin A to NBAG/NAD+ mixture, fragmentation of NAD+ and development of new peaks in HPLC chromatograms. Intracellular enzyme activity was confirmed by the prominent cytotoxic effects of exotoxin A extract on cultured cells. In conclusion, the activity of Exotoxin A can be monitored via its ADP-ribosyltransferase activity and low doses of gamma radiation reduced its activity. Therefore, coupling radiotherapy with exotoxin A in cancer therapy should be carefully monitored.

show abstract

“…Interestingly, pathogenic bacteria, such as Pseudomonas aeruginosa , have established routes for receptor-mediated endosomal uptake and retrograde transport into the cell cytoplasm [ 54 ]. The potential use of this pathway for cytoplasmic delivery while simultaneously addressing intracellular targets has only been described for a small set of cargo proteins [ 68 , 69 , 70 ]. Attempts have been made to induce cell-specific cytosol penetration through an endosomal uptake route, combined with the use of endosomal escape peptides (EEPs).…”

Section: Discussionmentioning

confidence: 99%

A Conditionally Activated Cytosol-Penetrating Antibody for TME-Dependent Intracellular Cargo Delivery

Dombrowsky,

Happel,

Habermann

et al. 2024

Antibodies

View full text Add to dashboard Cite

Currently, therapeutic and diagnostic applications of antibodies are primarily limited to cell surface-exposed and extracellular proteins. However, research has been conducted on cell-penetrating peptides (CPP), as well as cytosol-penetrating antibodies, to overcome these limitations. In this context, a heparin sulfate proteoglycan (HSPG)-binding antibody was serendipitously discovered, which eventually localizes to the cytosol of target cells. Functional characterization revealed that the tested antibody has beneficial cytosol-penetrating capabilities and can deliver cargo proteins (up to 70 kDa) to the cytosol. To achieve tumor-specific cell targeting and cargo delivery through conditional activation of the cell-penetrating antibody in the tumor microenvironment, a single-chain Fc fragment (scFv) and a VL domain were isolated as masking units. Several in vitro assays demonstrated that fusing the masking protein with a cleavable linker to the cell penetration antibody results in the inactivation of antibody cell binding and internalization. Removal of the mask via MMP-9 protease cleavage, a protease that is frequently overexpressed in the tumor microenvironment (TME), led to complete regeneration of binding and cytosol-penetrating capabilities. Masked and conditionally activated cytosol-penetrating antibodies have the potential to serve as a modular platform for delivering protein cargoes addressing intracellular targets in tumor cells.

show abstract

Intestinal Transcytosis of a Protein Cargo and Nanoparticles Mediated by a Non-Toxic Form of Pseudomonas aeruginosa Exotoxin A

Cited by 6 publications

References 27 publications

Zein-Based Nanoparticles as Oral Carriers for Insulin Delivery

Zein-Based Nanoparticles as Oral Carriers for Insulin Delivery

ADP-ribosyl transferase activity and gamma radiation cytotoxicity of Pseudomonas aeruginosa exotoxin A

A Conditionally Activated Cytosol-Penetrating Antibody for TME-Dependent Intracellular Cargo Delivery

Contact Info

Product

Resources

About