Gene optimized hantaan virus M segment DNA vaccine for hemorrhagic fever with renal syndrome
Inventors
Assignees
United States Department of the Army
Publication Number
US-9675684-B2
Publication Date
2017-06-13
Expiration Date
2033-03-28
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Abstract
A synthetic, codon-optimized Hantaan virus (HTNV) full-length M gene open reading frame that consists of a unique nucleotide sequence encoding HTNV proteins. This synthetic gene was cloned into a plasmid to form the first optimized HTNV full-length M gene that elicits neutralizing antibodies in animals when delivered in combination with a similarly optimized Puumala virus (PUUV) DNA vaccine. The invention obviates the need for an extraneous gene sequence that was previously required for expression of the non-optimized HTNV gene. The synthetic gene is engineered into a molecular vaccine system to prevent hemorrhagic fever with renal syndrome (HFRS) caused by infection with HTNV, SEOV, or DOBV. Alternatively, it can be combined with the optimized PUUV DNA vaccine to protect against HFRS caused by any hantavirus.
Core Innovation
The invention is a synthetic, codon-optimized Hantaan virus (HTNV) full-length M gene open reading frame (ORF) that encodes HTNV proteins. This synthetic gene is cloned into a plasmid to form a new optimized HTNV full-length M gene DNA vaccine that elicits neutralizing antibodies in animals when delivered in combination with a similarly optimized Puumala virus (PUUV) DNA vaccine. Unlike previous non-optimized HTNV DNA vaccines, the synthetic gene does not require extraneous gene sequences for expression.
The problem being solved is the gene-related immune interference that existed with prior HTNV vaccines when mixed with PUUV DNA vaccines. Previously, when the non-optimized HTNV DNA vaccine was mixed with the PUUV vaccine, neutralizing antibodies developed only against PUUV, hindering development of a comprehensive bivalent vaccine for hemorrhagic fever with renal syndrome (HFRS) caused by multiple hantaviruses. This interference limited protective immunity against all causative hantaviruses.
Through codon optimization to maximize mammalian expression, increased guanine-cytosine content for mRNA stability, and elimination of negative cis-acting motifs, the synthetic HTNV M gene DNA vaccine overcomes this interference problem. The invention enables administration of the HTNV vaccine mixed with an optimized PUUV DNA vaccine to elicit neutralizing antibodies against both viruses, thereby providing a safe, economical, flexible, and effective molecular vaccine system to prevent HFRS caused by HTNV, Seoul virus (SEOV), Dobrava virus (DOBV), and PUUV infections.
Claims Coverage
The patent contains multiple claims focusing on synthetic nucleic acids, DNA constructs, vectors, compositions, and formulations specifically comprising or encoding the optimized HTNV M gene sequence. There are seventeen claims, of which claims 1, 3, 6, and 11 are independent.
Synthetic nucleic acid comprising the optimized HTNV M gene
A synthetic nucleic acid comprising SEQ ID NO: 1, representing a codon-optimized, full-length HTNV M gene open reading frame optimized for mammalian expression without requiring extraneous gene sequences.
Vector and plasmid constructs comprising the optimized HTNV M gene
Vectors, including plasmids such as pWRG7077, comprising SEQ ID NO: 1 or SEQ ID NO: 2, which are synthetic DNA constructs encoding the codon-optimized HTNV M gene for use as DNA vaccines.
Immunogenic compositions and pharmaceutical formulations with the optimized HTNV M gene
Immunogenic compositions comprising SEQ ID NO: 1 or the synthetic DNA construct of SEQ ID NO: 2, formulated with pharmaceutically acceptable carriers, including compositions where DNA is delivered on gold particles or encapsulated in nanoparticles.
Recombinant DNA constructs with mammalian promoters
Recombinant DNA constructs comprising the optimized HTNV M gene cDNA sequence with functional mammalian promoters to enable expression of HTNV envelope glycoproteins (Gn and Gc).
The independent claims cover the synthetic, codon-optimized HTNV full-length M gene nucleic acid, its incorporation into vectors and plasmids for expression, immunogenic compositions and formulations for vaccination, and recombinant constructs including mammalian expression control elements. These claims protect the novel gene sequences, vector platforms, and vaccine formulations designed to overcome prior immune interference and enable combined vaccination against multiple hantaviruses.
Stated Advantages
Overcomes gene-related immune interference allowing simultaneous immunogenicity to HTNV and PUUV in a bivalent vaccine.
Does not require extraneous non-coding nucleotide sequences for expression, improving construct simplicity and efficiency.
Offers a safe, economical, flexible, and effective vaccine platform for preventing HFRS caused by multiple hantaviruses.
Compatible with advanced delivery methods such as intramuscular and intradermal electroporation, enhancing immune responses and reducing vaccine dose.
Facilitates mass vaccination due to improved stability, ease of manufacturing, and reduced cold-chain requirements.
Documented Applications
Prevention of hemorrhagic fever with renal syndrome (HFRS) caused by infection with Hantaan virus (HTNV), Seoul virus (SEOV), Dobrava virus (DOBV), and Puumala virus (PUUV).
Use as a DNA vaccine administered alone or in combination with an optimized PUUV DNA vaccine to elicit protective neutralizing antibodies to multiple hantaviruses.
Delivery via particle-mediated epidermal delivery (PMED), intramuscular electroporation (IM-EP), intradermal electroporation (ID-EP), jet injection, or nanoparticle encapsulation for enhanced immunogenicity.
Application in human clinical trials for active immunization against HFRS, including Phase 1 and Phase 2a studies evaluating safety, dosing, and immune responses.
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