Bluetongue Virus Particles as Nanoreactors for Enzyme Delivery and Cancer Therapy

Thuenemann, Eva C.; Le, Duc Anh; Lomonossoff, George P.; Steinmetz, Nicole F.

doi:10.1021/acs.molpharmaceut.0c01053

Cited by 18 publications

(22 citation statements)

References 33 publications

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“…The results presented in this study should allow us to extend and develop a set of plant VNPs for nanobody presentation. In order to avoid severe off-target effects of systemic drugs in medicine, several efforts are being made to develop targeted therapy using VNPs, in which drugs are either delivered specifically to tumor cells or activated specifically within them ( Shukla et al, 2020a ; Chung et al, 2020 ; Thuenemann et al, 2021 ). Therefore, further attempts in this area will involve the presentation of nanobodies with medicinal value.…”

Section: Discussionmentioning

confidence: 99%

Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants

Martí

Merwaiss

Butković

et al. 2022

Front. Bioeng. Biotechnol.

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Viral nanoparticles (VNPs) have recently attracted attention for their use as building blocks for novel materials to support a range of functions of potential interest in nanotechnology and medicine. Viral capsids are ideal for presenting small epitopes by inserting them at an appropriate site on the selected coat protein (CP). VNPs presenting antibodies on their surfaces are considered highly promising tools for therapeutic and diagnostic purposes. Due to their size, nanobodies are an interesting alternative to classic antibodies for surface presentation. Nanobodies are the variable domains of heavy-chain (VHH) antibodies from animals belonging to the family Camelidae, which have several properties that make them attractive therapeutic molecules, such as their small size, simple structure, and high affinity and specificity. In this work, we have produced genetically encoded VNPs derived from two different potyviruses—the largest group of RNA viruses that infect plants—decorated with nanobodies. We have created a VNP derived from zucchini yellow mosaic virus (ZYMV) decorated with a nanobody against the green fluorescent protein (GFP) in zucchini (Cucurbita pepo) plants. As reported for other viruses, the expression of ZYMV-derived VNPs decorated with this nanobody was only made possible by including a picornavirus 2A splicing peptide between the fused proteins, which resulted in a mixed population of unmodified and decorated CPs. We have also produced tobacco etch virus (TEV)-derived VNPs in Nicotiana benthamiana plants decorated with the same nanobody against GFP. Strikingly, in this case, VNPs could be assembled by direct fusion of the nanobody to the viral CP with no 2A splicing involved, likely resulting in fully decorated VNPs. For both expression systems, correct assembly and purification of the recombinant VNPs was confirmed by transmission electron microscope; the functionality of the CP-fused nanobody was assessed by western blot and binding assays. In sum, here we report the production of genetically encoded plant-derived VNPs decorated with a nanobody. This system may be an attractive alternative for the sustainable production in plants of nanobody-containing nanomaterials for diagnostic and therapeutic purposes.

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Section: Discussionmentioning

confidence: 99%

Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants

Martí

Merwaiss

Butković

et al. 2022

Front. Bioeng. Biotechnol.

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“…The Reoviruses, Rotavirus (RV) and Bluetongue virus (BTV) have also been used for encapsulation by translational fusion (Brillault et al, 2017;Charpilienne et al, 2001;Kar et al, 2005;Thuenemann et al, 2021). The VLP is a triple layer particle composed of 4 different proteins, although recombinant double layer core particles composed of two proteins and single layer sub-core particles composed of only one protein, are also possible.…”

Section: Translational Cp Fusionmentioning

confidence: 99%

“…The VLP is a triple layer particle composed of 4 different proteins, although recombinant double layer core particles composed of two proteins and single layer sub-core particles composed of only one protein, are also possible. The different particles exhibit a range of physical properties, with the core-like particles the most stable under physiological conditions (Jiménez-Zaragoza et al, 2018;Thuenemann et al, 2013), although the VLP possesses the most effective cell uptake ability (Thuenemann et al, 2021).…”

Section: Translational Cp Fusionmentioning

confidence: 99%

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Protein Cargo Encapsulation by Virus-Like Particles: Strategies and Applications

McNeale

Dashti

Cheah

et al. 2022

Preprint

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Viruses and the recombinant protein cages assembled from their structural proteins, known as virus-like particles (VLPs), have gain wide interest as tools in biotechnology and nanotechnology. Detailed structural information and their amenability to genetic and chemical modification make them attractive systems for further engineering. This review describes the range of non-enveloped viruses that have been co-opted for heterologous protein cargo encapsulation and the strategies that have been developed to drive encapsulation. Spherical capsids of a range of sizes have been used as platforms for protein cargo encapsulation. Various approaches, based on native and non-native interactions between the cargo proteins and inner surface of VLP capsids, have been devised to drive encapsulation. Here we outline the evolution of these approaches, discussing their benefits and limitations. Like the viruses from which they are derived, VLPs are of interest in both biomedical and materials applications. The encapsulation of protein cargo inside VLPs leads to numerous uses in both fundamental and applied biocatalysis and biomedicine, some of which are discussed herein. The applied science of protein encapsulating VLPs is emerging as a research field with great potential. Developments in loading control, higher order assembly and capsid optimization are poised to realize this potential in the near future.

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“…Reverse genetics systems for BTV were developed based on in vitro transcribed RNA ( Boyce et al, 2008 ) and expression plasmids ( Pretorius et al, 2015 ) with applications in nanotechnology and vaccine development ( Roy, 2020 ; Thuenemann et al, 2021 ). The particle-to-pfu ratio for BTV infection of mammalian cells is reported to be very low (1–3; Wu et al, 2019 ; Labadie and Roy, 2020 ) compared to other viruses, illustrating high infection efficiency.…”

Section: Molecular Mechanism Of Cellular Entry By Btv (Genus: Orbivirus ) In Mammalian and Insect Cellsmentioning

confidence: 99%

Mechanisms of Cell Entry by dsRNA Viruses: Insights for Efficient Delivery of dsRNA and Tools for Improved RNAi-Based Pest Control

et al. 2021

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While RNAi is often heralded as a promising new strategy for insect pest control, a major obstacle that still remains is the efficient delivery of dsRNA molecules within the cells of the targeted insects. However, it seems overlooked that dsRNA viruses already have developed efficient strategies for transport of dsRNA molecules across tissue barriers and cellular membranes. Besides protecting their dsRNA genomes in a protective shell, dsRNA viruses also display outer capsid layers that incorporate sophisticated mechanisms to disrupt the plasma membrane layer and to translocate core particles (with linear dsRNA genome fragments) within the cytoplasm. Because of the perceived efficiency of the translocation mechanism, it is well worth analyzing in detail the molecular processes that are used to achieve this feat. In this review, the mechanism of cell entry by dsRNA viruses belonging to the Reoviridae family is discussed in detail. Because of the large amount of progress in mammalian versus insect models, the mechanism of infections of reoviruses in mammals (orthoreoviruses, rotaviruses, orbiviruses) will be treated as a point of reference against which infections of reoviruses in insects (orbiviruses in midges, plant viruses in hemipterans, insect-specific cypoviruses in lepidopterans) will be compared. The goal of this discussion is to uncover the basic principles by which dsRNA viruses cross tissue barriers and translocate their cargo to the cellular cytoplasm; such knowledge subsequently can be incorporated into the design of dsRNA virus-based viral-like particles for optimal delivery of RNAi triggers in targeted insect pests.

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Bluetongue Virus Particles as Nanoreactors for Enzyme Delivery and Cancer Therapy

Cited by 18 publications

References 33 publications

Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants

Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants

Protein Cargo Encapsulation by Virus-Like Particles: Strategies and Applications

Mechanisms of Cell Entry by dsRNA Viruses: Insights for Efficient Delivery of dsRNA and Tools for Improved RNAi-Based Pest Control

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