Virus-like particles and methods of use
Inventors
Nabel, Gary J. • Rao, Srinivas • Akahata, Wataru
Assignees
US Department of Health and Human Services
Publication Number
US-11098084-B2
Publication Date
2021-08-24
Expiration Date
2032-01-31
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Abstract
The invention features modified alphavirus or flavivirus virus-like particles (VLPs). The invention provides methods, compositions, and kits featuring the modified VLPs. The invention also features methods for enhancing production of modified VLPs for use in the prevention or treatment of alphavirus and flavivirus-mediated diseases. The invention also provides methods for delivering agents to a cell using the modified VLPs.
Core Innovation
The invention features modified alphavirus or flavivirus virus-like particles (VLPs), including alterations that enhance or allow VLP production, particularly in the E2 protein or the alphavirus capsid protein Nuclear Localization Signal (NLS). Methods, compositions, and kits utilizing these modified VLPs are provided, along with methods for enhancing production of these VLPs to be used in prevention or treatment of diseases mediated by alphaviruses and flaviviruses. Additionally, the invention addresses methods for delivering agents to cells using these modified VLPs.
The problem addressed by the invention arises from the serious public health threat posed by alphaviruses and flaviviruses, which cause diseases ranging from encephalitis to severe hemorrhages and arthritis, without vaccines or antiviral therapies currently available. Production of VLPs is challenging, as expression of wild-type alphavirus proteins does not always produce VLPs efficiently, exemplified by Eastern equine encephalitis virus (EEEV) and Western equine encephalitis virus (WEEV) strains. Similarly, flavivirus VLP production can be inefficient.
The invention overcomes these issues by identifying specific amino acid alterations that enhance VLP production, such as substitution at amino acid position 234 in the alphavirus E2 protein (e.g., lysine to asparagine) and modifications at position 251 that destabilize the E2 protein during viral budding. Moreover, alterations in the charged residues of the alphavirus capsid protein NLS reduce nuclear localization, allowing increased VLP secretion. This enhances yields of VLPs, facilitating their use as safe and immunogenic vaccines and as delivery vehicles for agents into cells.
Claims Coverage
The patent claims cover several inventive features related to immunogenic compositions comprising virus-like particles with specific protein alterations, methods of inducing immune responses using such VLPs, and methods of producing VLPs with enhanced yields.
Modification of alphavirus E2 protein to enhance VLP production
An alphavirus E2 protein comprising one or more alterations at amino acid locations corresponding to positions 170, 200, 233, 234, 251, or 256 of CHIKV E2 protein, where such alterations enhance VLP production.
Alteration of alphavirus capsid protein nuclear localization signal residues
An alphavirus capsid protein comprising one or more alterations at charged amino acid residues within the Nuclear Localization Signal (NLS), specifically targeting regions of amino acids 67-70 for EEEV and WEEV capsid proteins, 64-68 for VEEV and Barmah Forest virus capsid proteins, 62-69 for CHIKV capsid protein, and 71-74 for Ross River virus capsid protein, where such alterations enhance VLP production.
Immunogenic compositions comprising VLPs with specified protein alterations
Compositions comprising VLPs self-assembled from alphavirus E2 proteins with the specified amino acid alterations and/or alphavirus capsid proteins with altered NLS charged residues, formulated to induce immune responses against alphaviruses.
Methods of inducing immune responses to alphaviruses using altered VLPs
Administration of immunogenic compositions comprising VLPs with altered alphavirus E2 proteins and/or capsid proteins as described, to induce protective immune responses in individuals against alphavirus infections.
Methods of producing VLPs with enhanced yields by expressing altered alphavirus structural proteins
Methods involving expression of alphavirus E2 proteins with specified amino acid alterations and/or alphavirus capsid proteins with altered NLS charged residues in cells, resulting in the formation of VLPs with enhanced production.
The claims broadly cover immunogenic compositions containing VLPs with specific E2 protein and capsid protein NLS alterations that enhance VLP production, methods of inducing immune responses using these VLPs, and methods for their production. These inventive features collectively enable improved production and immunogenicity of alphavirus VLP-based vaccines and related compositions.
Stated Advantages
Enhanced production yields of alphavirus and flavivirus VLPs through specific amino acid alterations.
Improved safety and immunogenicity of VLP vaccines due to their virus-like structure but non-infectious nature.
Ability to induce potent neutralizing antibody responses against homologous and heterologous alphavirus strains.
Protection against viremia and inflammatory consequences of alphavirus infection shown in animal models.
Use of VLPs as delivery vehicles for agents to cells, expanding therapeutic and diagnostic applications.
Documented Applications
Prevention or treatment of alphavirus-mediated diseases such as those caused by Chikungunya virus, Eastern equine encephalitis virus, Western equine encephalitis virus, Venezuelan equine encephalitis virus, Ross River virus, and Barmah Forest virus.
Prevention or treatment of flavivirus-mediated diseases including disease caused by Yellow Fever Virus, Dengue Virus, Japanese Encephalitis Virus, Tick-Borne Encephalitis Virus, and West Nile Virus.
Use of VLPs as vaccines to induce protective immune responses in subjects against alphaviruses and flaviviruses.
Use of VLPs for delivering therapeutic, diagnostic, or imaging agents into target cells.
Methods of producing VLPs with improved yield for vaccine manufacture and therapeutic use.
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