Brome mosaic virus-like particles as siRNA nanocarriers for biomedical purposes

Núñez-Rivera, Alfredo; Fournier, Pierrick G.J.; Arellano, Danna L.; Rodríguez-Hernández, Ana G.; Vázquez-Duhalt, Rafael; Cadena-Nava, Rubén D.

doi:10.3762/bjnano.11.28

Cited by 37 publications

(33 citation statements)

References 60 publications

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“…The use of a fluorescent labelling is necessary to detect the VLP's by confocal microscopy. In a previous work we demonstrated that the VLP's from BMV are able to be internalized with and without fluorophore in tumor cells, [42] as well other groups demonstrated the cell internalization VLP′s from BMV into bronchial epithelial cells [44] . Hepatocytes, fibroblasts and kidney cells showed abundant cell internalization as demonstrated by the green fluorescence of FITC.…”

Section: Resultsmentioning

confidence: 61%

“…Here, GALT has been effectively encapsulated in a VLP form BMV (Figures 3 and 4), preserving catalytic activity (Table 1). Virus‐like nanoparticles from the plant virus BMV seems to be a good candidate for the enzyme delivery because it shows a very low immunogenicity [42] …”

Section: Discussionmentioning

confidence: 99%

“…Capsid protein obtention and purification . Brome mosaic virions (BMV) were purified from infected barley plants ( Hordeum vulgare ) as previously described [42] . Briefly, after four weeks′ germination, plants were infected by leaf abrasion with 10 μL of inoculum consisted of 0.1 μg μL −1 native virus in a 10 mM sodium phosphate buffer, pH 6, containing 10 mM MnCl 2 .…”

Section: Methodsmentioning

confidence: 99%

“…Nanoreactor synthesis . Galactose‐1‐phosphate uridyl transferase (GALT) was encapsulated inside BMV VLPs by trapping taking advantage of the electrostatic interactions and following previously reported procedure for the encapsulation of RNA [42] . Different GALT/CP ratios were assayed.…”

Section: Methodsmentioning

confidence: 99%

“…This encapsulated fluorescent dye‐binding aptamer extends time single particle tracking due to negligible bleaching from continuous chromophore exchange, which makes them potentially useful for studies of in vivo virus dynamics. Finally, small interfering RNA (siRNA) has recently encapsulated in BMV VLP′s [42] . The gene silencing was demonstrated by VLPs loaded with siGFP and tested on cells that constitutively express the green fluorescent protein (GPF).…”

Section: Introductionmentioning

confidence: 99%

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Virus‐Based Nanoreactors with GALT Activity for Classic Galactosemia Therapy

et al. 2021

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Enzymatic nanoreactors were obtained by galactose‐1‐phosphate uridylyl‐transferase (GALT) encapsulation into plant virus capsids by a molecular self‐assembly strategy. The aim of this work was to produce virus‐like nanoparticles containing GALT for an enzyme‐replacement therapy for classic galactosemia. The encapsulation efficiency and the catalytic constants of bio‐nanoreactors were determined by using different GALT and virus coat protein ratios. The substrate affinity of nanoreactors was slightly lower than that of the free enzyme; the activity rate was 16 % of the GALT free enzyme. The enzymatic nanoreactors without functionalization were internalized into different cell lines including fibroblast and kidney cells, but especially into hepatocytes. The enzymatic nanoreactors are an innovative enzyme preparation with potential use for the treatment of classic galactosemia.

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Section: Resultsmentioning

confidence: 61%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Virus‐Based Nanoreactors with GALT Activity for Classic Galactosemia Therapy

et al. 2021

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Glucose oxidase virus‐based nanoreactors for smart breast cancer therapy

Gama

Juàrez

Rodríguez-Hernández

et al. 2023

Biotechnology Journal

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BackgroundBreast cancer is the most common malignant tumor disease and the leading cause of female mortality. The evolution of nanomaterials science opens the opportunity to improve traditional cancer therapies, enhancing therapy efficiency and reducing side effects.Methods and major resultsHerein, protein cages conceived as enzymatic nanoreactors were designed and produced by using virus‐like nanoparticles (VLPs) from Brome mosaic virus (BMV) and containing the catalytic activity of glucose oxidase (GOx) enzyme. The GOx enzyme was encapsulated into the BMV capsid (VLP‐GOx), and the resulting enzymatic nanoreactors were coated with human serum albumin (VLP‐GOx@HSA) for breast tumor cell targeting. The effect of the synthesized GOx nanoreactors on breast tumor cell lines was studied in vitro. Both nanoreactor preparations VLP‐GOx and VLP‐GOx@HSA showed to be highly cytotoxic for breast tumor cell cultures. Cytotoxicity for human embryonic kidney cells was also found. The monitoring of nanoreactor treatment on triple‐negative breast cancer cells showed an evident production of oxygen by the catalase antioxidant enzyme induced by the high production of hydrogen peroxide from GOx activity.Conclusions and implicationsThe nanoreactors containing GOx activity are entirely suitable for cytotoxicity generation in tumor cells. The HSA functionalization of the VLP‐GOx nanoreactors, a strategy designed for selective cancer targeting, showed no improvement in the cytotoxic effect. The GOx containing enzymatic nanoreactors seems to be an interesting alternative to improve the current cancer therapy. In vivo studies are ongoing to reinforce the effectiveness of this treatment strategy.

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Targeted Enzymatic VLP‐Nanoreactors with β‐Glucocerebrosidase Activity as Potential Enzyme Replacement Therapy for Gaucher's Disease

et al. 2022

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Gaucher disease is a genetic disorder and the most common lysosomal disease caused by the deficiency of enzyme βglucocerebrosidase (GCase). Although enzyme replacement therapy (ERT) is successfully applied using mannose-exposed conjugated glucocerebrosidase, the lower stability of the enzyme in blood demands periodic intravenous administration that adds to the high cost of treatment. In this work, the enzyme β-glucocerebrosidase was encapsulated inside viruslike nanoparticles (VLPs) from brome mosaic virus (BMV), and their surface was functionalized with mannose groups for targeting to macrophages. The VLP nanoreactors showed significant GCase catalytic activity. Moreover, the Michaelis-Menten constants for the free GCase enzyme (K M = 0.29 mM) and the functionalized nanoreactors (K M = 0.32 mM) were similar even after chemical modification. Importantly, the stability of enzymes under physiological conditions (pH 7.4, 37 °C) was enhanced by � 11-fold after encapsulation; this is beneficial for obtaining a higher blood circulation half-life, which may decrease the cost of therapy by reducing the requirement of multiple intravenous injections. Finally, the mannose receptor targeted enzymatic nanoreactors showed enhanced internalization into macrophage cells. Thus, the catalytic activity and cell targeting suggest the potential of these nanoreactors in ERT of Gaucher's disease.

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Brome mosaic virus-like particles as siRNA nanocarriers for biomedical purposes

Cited by 37 publications

References 60 publications

Virus‐Based Nanoreactors with GALT Activity for Classic Galactosemia Therapy

Virus‐Based Nanoreactors with GALT Activity for Classic Galactosemia Therapy

Glucose oxidase virus‐based nanoreactors for smart breast cancer therapy

Targeted Enzymatic VLP‐Nanoreactors with β‐Glucocerebrosidase Activity as Potential Enzyme Replacement Therapy for Gaucher's Disease

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