Figure 1 - uploaded by Prachi Anand
Content may be subject to copyright.
Trojan horse virus like particle design for delivering peptides across the BBB. Concept figure of engineered viral nanocontainers encapsulating marine snail peptide MVIIA in the interior and cell penetrating peptide Tat(FAM) on the exterior shuttle the nanocontainers across the BBB using an endocytic pathway.
Source publication
The blood brain barrier (BBB) is often an insurmountable obstacle for a large number of candidate drugs, including peptides, antibiotics, and chemotherapeutic agents. Devising an adroit delivery method to cross the BBB is essential to unlocking widespread application of peptide therapeutics. Presented here is an engineered nanocontainer for deliver...
Context in source publication
Context 1
... analysis of preheated P22-MVIIA capsids revealed a 10, 267.9 Da mass for the MVIIA-SP238 fusion protein, indicating that the MVIIA peptide was in a folded conformation. The observed mass of 10, 267.9 Da is consistent with a potassium adduct of the MVIIA-SP238 fusion protein, with the loss of six hydrogen atoms, corresponding to the formation of 3 disulfide bonds in the MVIIA peptide ( Supplementary Fig. 1). While it is not possible to determine if the MVIIA expressed as a protein fusion is in its native form, pre- vious attempts at recombinant expression and oxidative folding experiments of chemically synthesized cone snail neuropeptides have been shown to fold into native conformations 29 . ...
Citations
... Efforts are underway to enhance the pharmaceutical properties of ziconotide for intravenous, oral, or transdermal administration to broaden the therapeutic window and simplify its use [55,128]. Subtle structural modifications have mainly focused on preserving the overall structure of ziconotide. ...
... Myristoylated N-terminal ziconotide can undergo self-assembly onto micelles, resulting in extended analgesic effects and a notable reduction in the risk of side effects such as tremors and impaired motor coordination in mice [104]. Numerous patents have been filed to theoretically modify ziconotide's structure to enhance its ability to cross the BBB, but none of these modifications have been validated in humans or made commercially available [128,134]. Leconotide, however, when administered non-intrathecally, was denied approval due to the serious complications that emerged during phase II clinical trials [135]. ...
Managing severe chronic pain is a challenging task, given the limited effectiveness of available pharmacological and non-pharmacological treatments. This issue continues to be a significant public health concern, requiring a substantial therapeutic response. Ziconotide, a synthetic peptide initially isolated from Conus magus in 1982 and approved by the US Food and Drug Administration and the European Medicines Agency in 2004, is the first-line intrathecal method for individuals experiencing severe chronic pain refractory to other therapeutic measures. Ziconotide produces powerful analgesia by blocking N-type calcium channels in the spinal cord, which inhibits the release of pain-relevant neurotransmitters from the central terminals of primary afferent neurons. However, despite possessing many favorable qualities, including the absence of tolerance development, respiratory depression, and withdrawal symptoms (largely due to the absence of a G-protein mediation mechanism), ziconotide's application is limited due to factors such as intrathecal administration and a narrow therapeutic window resulting from significant dose-related undesired effects of the central nervous system. This review aims to provide a comprehensive and clinically relevant summary of the literatures concerning the pharmacokinetics and metabolism of intrathecal ziconotide. It will also describe strategies intended to enhance clinical efficacy while reducing the incidence of side effects. Additionally, the review will explore the current efforts to refine the structure of ziconotide for better clinical outcomes. Lastly, it will prospect potential developments in the new class of selective N-type voltage-sensitive calcium-channel blockers.
... In a study conducted by Anand et al., Salmonella typhimurium (S. typhimurium) bacteriophage P22 was genetically engineered to express a peptide on its capsid that enables the phage to pass through the BBB. Results of this showed that the Ziconotide peptide is well expressed in snail venom and has analgesic properties [92]. Shiga-like toxin produced by EHEC causes infammation in the intestine, resulting in an increase in proinfammatory cytokines such as IL-6 as well as IgG1, IgG2a, and IgA levels due to the activation of the immune system. ...
The emergence of antibiotic-resistant strains, the decreased effectiveness of conventional therapies, and the side effects have led researchers to seek a safer, more cost-effective, patient-friendly, and effective method that does not develop antibiotic resistance. With progress in synthetic biology and genetic engineering, genetically engineered microorganisms effective in treatment, prophylaxis, drug delivery, and diagnosis have been developed. The present study reviews the types of genetically engineered bacteria and phages, their impacts on diseases, cancer, and metabolic and inflammatory disorders, the biosynthesis of these modified strains, the route of administration, and their effects on the environment. We conclude that genetically engineered microorganisms can be considered promising candidates for adjunctive treatment of diseases and cancers.
... Not only did the phages bind to eucaryotic cells, but they were also internalized into the cells through endocytosis and largely retained infectivity for 24 h [219]. Anand et al. developed a bifunctional viral nanocontainer based on the Salmonella-specific bacteriophage P22 [31]. Ziconotide, an analgesic peptide drug derived from a marine snail, was incorporated into the nanocontainer, while a cell-penetrating peptide HIV-tat was displayed on the exterior surface of the phage capsid. ...
... Such a modified P22 was successfully transferred into several cell-based models of rat brain microvascular endothelial cells as well as human brain microvascular endothelial cells. Hence, these findings open another alternative route for drug administration [31]. The F88 phage was also reported to easily penetrate the BBB after intranasal administration. ...
The central nervous system manages all of our activities (e.g., direct thinking and decision-making processes). It receives information from the environment and responds to environmental stimuli. Bacterial viruses (bacteriophages, phages) are the most numerous structures occurring in the biosphere and are also found in the human organism. Therefore, understanding how phages may influence this system is of great importance and is the purpose of this review. We have focused on the effect of natural bacteriophages in the central nervous system, linking them to those present in the gut microbiota, creating the gut-brain axis network, as well as their interdependence. Importantly, based on the current knowledge in the field of phage application (e.g., intranasal) in the treatment of bacterial diseases associated with the brain and nervous system, bacteriophages may have significant therapeutic potential. Moreover, it was indicated that bacteriophages may influence cognitive processing. In addition, phages (via phage display technology) appear promising as a targeted therapeutic tool in the treatment of, among other things, brain cancers. The information collected and reviewed in this work indicates that phages and their impact on the nervous system is a fascinating and, so far, underexplored field. Therefore, the aim of this review is not only to summarize currently available information on the association of phages with the nervous system, but also to stimulate future studies that could pave the way for novel therapeutic approaches potentially useful in treating bacterial and non-bacterial neural diseases.
... Quantitative densitometry revealed that 24-27 anti-PD-1 peptides were linked onto each CPMV VLP. The anti-PD-1-decorted CPMV VLPs presented increased antitumor efficacy in the tumor mouse model [41]. If there is no exposed cysteine on the outer surface of PNCs, it will be convenient to sitespecifically introduce cysteines with controlled numbers on demand. ...
Background
As one of the representative protein materials, protein nanocages (PNCs) are self‐assembled supramolecular structures with multiple advantages, such as good monodispersity, biocompatibility, structural addressability, and facile production. Precise quantitative functionalization is essential to the construction of PNCs with designed purposes.
Results
With three modifiable interfaces, the interior surface, outer surface, and interfaces between building blocks, PNCs can serve as an ideal platform for precise multi‐functionalization studies and applications. This review summarizes the currently available methods for precise quantitative functionalization of PNCs and highlights the significance of precise quantitative control in fabricating PNC‐based materials or devices. These methods can be categorized into three groups, genetic, chemical, and combined modification.
Conclusion
This review would be constructive for those who work with biosynthetic PNCs in diverse fields.
... The surface of VLPs can be functionalized with other biomolecules for stratified targeting in forms of genetic fusion [116]. For instance, the HIV-Tat-labeled P22 VLP can readily package a drug-scaffold fusion protein and transport it across rat and human BEC models [117]. In contrast, the MS2 VLP requires a 19-nucleotide stem loop (the pac site) to efficiently package mRNA [118], protein [119], siRNA [120], antisense oligonucleotide [121], and microRNA [122]. ...
Nanomaterials offer the potential for positive technological impact in a variety of industries. The major breakthrough is in neurological therapeutic applications as their physical and chemical properties allow them to penetrate the blood-brain barrier (BBB). However, questions concerning its safety have arisen as a result of its permeability and the broad application of nanomaterials especially the engineered nanomaterial (ENMs). Due to the large spectrum of ENM properties, pinpointing individual features that caused toxicity is difficult. It is therefore urgent to capitalise on these new developments in ENM safety evaluation. Indeed, novel risk assessment and risk management techniques for humans and the environment across the whole life-cycle of nanomaterial products have emerged in recent years, including systems biology approaches and high-throughput screening platforms. Moreover, the new toxicology technology should practically reduce the number of animal samples required for testing and allow both in vitro and in vivo cell studies. Unlike traditional cytotoxicity, which limits the analysis effect to a single experiment, hazardous risk assessment by integrated omics technologies using high-throughput technologies provides robustness of systemic functional analysis towards ENM, allowing the discovery of biomarkers and functional pathways affecting ENM safety application.
... As mentioned earlier, a large segment of the SP sequence can be changed, and its N-terminus can be considerably truncated with no major consequences to capsid assembly or structure. Previous works have exploited this aspect of the VLP for the in vivo packaging of cargo utilizing truncated variants of SP composed of amino acids 141-303 and 238-303 [16,17,56]. The truncated SP has different properties and effects on the capsid. ...
... This promotes nonspecific interactions and the retention of the SP inside the capsid [13]. The use of this truncated variant also grants a better utilization of the VLP's internal space, allowing the packaging of a higher number of protein fusions in the capsid [56]. This way, exogenous proteins can be genetically fused to SP [13,16,57], and other molecules can be encapsulated during the assembly or through the pores after the particle has formed [9]. ...
... 97-130 and situated on the P-domain, is oriented toward the interior of the capsid, and its mutations did not affect the capsid structure, making it a good site for internal modifications [19]. Several residues were already substituted to be used as binding sites for the incorporation of exogenous structures by chemical modifications, e.g., peptides and drugs [9,12,19,56]. Both cargo-loading strategies and particle structural modifications will be explored later. ...
The Salmonella enterica bacteriophage P22 is one of the most promising models for the development of virus-like particle (VLP) nanocages. It possesses an icosahedral T = 7 capsid, assembled by the combination of two structural proteins: the coat protein (gp5) and the scaffold protein (gp8). The P22 capsid has the remarkable capability of undergoing structural transition into three morphologies with differing diameters and wall-pore sizes. These varied morphologies can be explored for the design of nanoplatforms, such as for the development of cargo internalization strategies. The capsid proteic nature allows for the extensive modification of its structure, enabling the addition of non-native structures to alter the VLP properties or confer them to diverse ends. Various molecules were added to the P22 VLP through genetic, chemical, and other means to both the capsid and the scaffold protein, permitting the encapsulation or the presentation of cargo. This allows the particle to be exploited for numerous purposes—for example, as a nanocarrier, nanoreactor, and vaccine model, among other applications. Therefore, the present review intends to give an overview of the literature on this amazing particle.
... However, phage engineering has made it possible to design a delivery shuttle for drug cargoes across the blood-brain barriers by using the Trojan horse strategy. The human immunodeficiency virus type-1 has been used as a penetrating peptide in the cell to the exterior of phages P22 and successfully delivered the neuropeptide zicotide in a snail in vivo bloodbrain barrier model (Anand et al., 2015). Based on the above studies, it can be concluded that nanoparticles-based on phages could cross the blood-brain barrier for specific drug cargo delivery, diagnosis, and therapeutic strategies for treating brain disorders like Alzheimer's disease. ...
Bacteriophages are the most abundant biological entity in the world and hold a tremendous amount of unexplored genetic information. Since their discovery, phages have drawn a great deal of attention from researchers despite their small size. The development of advanced strategies to modify their genomes and produce engineered phages with desired traits has opened new avenues for their applications. This review presents advanced strategies for developing engineered phages and their potential antibacterial applications in phage therapy, disruption of biofilm, delivery of antimicrobials, use of endolysin as an antibacterial agent, and altering the phage host range. Similarly, engineered phages find applications in eukaryotes as a shuttle for delivering genes and drugs to the targeted cells, and are used in the development of vaccines and facilitating tissue engineering. The use of phage display-based specific peptides for vaccine development, diagnostic tools, and targeted drug delivery is also discussed in this review. The engineered phage-mediated industrial food processing and biocontrol, advanced wastewater treatment, phage-based nano-medicines, and their use as a bio-recognition element for the detection of bacterial pathogens are also part of this review. The genetic engineering approaches hold great potential to accelerate translational phages and research. Overall, this review provides a deep understanding of the ingenious knowledge of phage engineering to move them beyond their innate ability for potential applications.
... The peptide was encased in a viral container and administered across the BBB using a trojan horse approach, which has previously been described as a viable replacement for intrathecal injection in order to deliver ziconotide. [128] An strategy for inducing disintegration of a nano container made from the viral capsid of the Salmonella typhimurium bacteriophage P22 is disclosed in physiologic conditions. The P22 capsid is an icosahedral lattice that self-assembles when enough copies of the P22 scaffold protein are present. ...
Conventional drug delivery systems are challenged by concerns related to systemic toxicity, repetitive doses, drug concentrations fluctuation, and adverse effects. Various drug delivery systems are developed to overcome these limitations. Nanomaterials are employed in a variety of biomedical applications such as therapeutics delivery, cancer therapy, and tissue engineering. Physiochemical nanoparticle assembly techniques involve the application of solvents and potentially harmful chemicals, commonly at high temperatures. Genetically engineered organisms have the potential to be used as promising candidates for greener, efficient, and more adaptable platforms for the synthesis and assembly of nanomaterials. Genetically engineered carriers are precisely designed and constructed in shape and size, enabling precise control over drug attachment sites. The high accuracy of these novel advanced materials, biocompatibility, and stimuli‐responsiveness, elucidate their emerging application in controlled drug delivery. The current article represents the research progress in developing various genetically engineered carriers. Organic‐based nanoparticles including cellulose, collagen, silk‐like polymers, elastin‐like protein, silk‐elastin‐like protein, and inorganic‐based nanoparticles are discussed in detail. Afterward, viral‐based carriers are classified, and their potential for targeted therapeutics delivery is highlighted. Finally, the challenges and prospects of these delivery systems are concluded.
... Attached on the exterior of the nanocontainer More than one uptake mechanism via receptor-mediated endocytic pathways [136]. ...
Central nervous system (CNS) diseases are the leading causes of death and disabilities in the world. It is quite challenging to treat CNS diseases efficiently because of the blood–brain barrier (BBB). It is a physical barrier with tight junction proteins and high selectivity to limit the substance transportation between the blood and neural tissues. Thus, it is important to understand BBB transport mechanisms for developing novel drug carriers to overcome the BBB. This paper introduces the structure of the BBB and its physiological transport mechanisms. Meanwhile, different strategies for crossing the BBB by using nanomaterial-based drug carriers are reviewed, including carrier-mediated, adsorptive-mediated, and receptor-mediated transcytosis. Since the viral-induced CNS diseases are associated with BBB breakdown, various neurotropic viruses and their mechanisms on BBB disruption are reviewed and discussed, which are considered as an alternative solution to overcome the BBB. Therefore, most recent studies on virus-mimicking nanocarriers for drug delivery to cross the BBB are also reviewed and discussed. On the other hand, the routes of administration of drug-loaded nanocarriers to the CNS have been reviewed. In sum, this paper reviews and discusses various strategies and routes of nano-formulated drug delivery systems across the BBB to the brain, which will contribute to the advanced diagnosis and treatment of CNS diseases.
... Phages have successfully been designed to shuttle drug cargos across the blood brain barrier using Trojan horse strategies. For example, by conjugating a cell penetrating peptide from the Tat protein of human immunodeficiency virus type-1 to the exterior of P22 phage particles carrying the snail neuropeptide ziconotide, Anand and colleagues demonstrated transportation of ziconotide in several in vitro blood-brain barrier models [87]. Apawu and colleagues conjugated the synthetic peptide angiopep-2 to the capsid of MS2 containing an MRI detectable Mn 2+ coordinated porphyrin ring and demonstrated these nanoparticles crossed the blood-brain barrier in rats [88]. ...
Since their independent discovery by Frederick Twort in 1915 and Felix d’Herelle in 1917, bacteriophages have captured the attention of scientists for more than a century. They are the most abundant organisms on the planet, often outnumbering their bacterial hosts by tenfold in a given environment, and they constitute a vast reservoir of unexplored genetic information. The increased prevalence of antibiotic resistant pathogens has renewed interest in the use of naturally obtained phages to combat bacterial infections, aka phage therapy. The development of tools to modify phages, genetically or chemically, combined with their structural flexibility, cargo capacity, ease of propagation, and overall safety in humans has opened the door to a myriad of applications. This review article will introduce readers to many of the varied and ingenious ways in which researchers are modifying phages to move them well beyond their innate ability to target and kill bacteria.
