Zoonotic disease rna vaccines

Abstract

The disclosure relates to Lassa virus, Nipah virus, and betacoronavirus ribonucleic acid vaccines as well as methods of using the vaccines and compositions comprising the vaccines.

What is claimed is: 
     
         1 . A zoonotic disease vaccine, comprising a ribonucleic acid (RNA) comprising an open reading frame (ORF) encoding an antigen selected from Lassa virus antigens, Nipah virus antigens, and betacoronavirus antigens, wherein intramuscular (IM) administration of a therapeutically effective amount of the vaccine to a subject induces an immune response in the subject. 
     
     
         2 . The zoonotic disease vaccine of  claim 1 , wherein the ORF encodes a Lassa virus antigen. 
     
     
         3 . The zoonotic disease vaccine of  claim 2 , wherein the Lassa virus antigen comprises a glycoprotein. 
     
     
         4 . The zoonotic disease vaccine of  claim 3 , wherein the Lassa virus antigen comprises a Lassa virus glycoprotein precursor (GPC), a structurally stabilized Lassa virus GPC, an ectodomain of Lassa virus glycoprotein 1 (GP1), or a Lassa virus glycoprotein 2 (GP2). 
     
     
         5 . The zoonotic disease vaccine of  claim 4 , wherein the Lassa virus antigen comprises amino acid residues 59-259 of a Lassa virus GPC. 
     
     
         6 . The zoonotic disease vaccine of  claim 2 , wherein the Lassa virus antigen comprises a nucleocapsid protein (NP). 
     
     
         7 . The zoonotic disease vaccine of  claim 2 , wherein the Lassa virus antigen has an amino acid sequence that has at least 90%, at least 95%, or at least 99% identity to an amino acid sequence identified by any one of SEQ ID NO: 1-3, but does not include wild-type protein sequence. 
     
     
         8 . The zoonotic disease vaccine of  claim 2 , wherein the Lassa virus antigen has an amino acid sequence of any one of SEQ ID NO: 1-3. 
     
     
         9 . The zoonotic disease vaccine of  claim 2 , wherein the RNA comprising an ORF sequence has at least 90%, at least 95%, or at least 99% identity to a nucleic acid sequence identified by any one of SEQ ID NO: 6, 7 or 9, but does not include wild-type protein sequence. 
     
     
         10 . The zoonotic disease vaccine of  claim 2 , wherein the RNA comprising an ORF sequence comprises a nucleic acid sequence of any one of SEQ ID NO: 6, 7 or 9. 
     
     
         11 . The zoonotic disease vaccine of  claim 1 , wherein the ORF encodes a Nipah virus antigen and/or a Hendra virus antigen. 
     
     
         12 . The zoonotic disease vaccine of  claim 11 , wherein the Nipah virus antigen and/or a Hendra virus antigen comprises a hemagglutinin-neuraminidase protein (HN), a hemagglutinin protein (H), or a glycoprotein (G). 
     
     
         13 . The zoonotic disease vaccine of  claim 12 , wherein the Nipah virus antigen and/or a Hendra virus antigen comprises an attachment glycoprotein, optionally a type II membrane protein. 
     
     
         14 . The zoonotic disease vaccine of  claim 12 , wherein the Nipah virus antigen and/or a Hendra virus antigen comprises a fusion (F) glycoprotein. 
     
     
         15 . The zoonotic disease vaccine of  claim 14 , wherein the F glycoprotein comprises a trimeric class I fusogenic envelope glycoprotein containing two heptad repeat (HR) regions and a hydrophobic fusion peptide. 
     
     
         16 . The zoonotic disease vaccine of any one of  claims 11 - 15 , wherein the Nipah virus antigen and/or a Hendra virus antigen is a Nipah virus antigen. 
     
     
         17 . The zoonotic disease vaccine of any one of  claims 11 - 15 , wherein the Nipah virus antigen and/or a Hendra virus antigen is a Hendra virus antigen. 
     
     
         18 . The zoonotic disease vaccine of  claim 11 , wherein the Nipah virus antigen and/or a Hendra virus antigen has an amino acid sequence that has at least 90%, at least 95%, or at least 99% identity to an amino acid sequence identified by any one of SEQ ID NO: 10-13 but does not include wild-type protein sequence. 
     
     
         19 . The zoonotic disease vaccine of  claim 11 , wherein the Nipah virus antigen and/or a Hendra virus antigen has an amino acid sequence of any one of SEQ ID NO: 10-13. 
     
     
         20 . The zoonotic disease vaccine of  claim 11 , wherein the RNA comprising an ORF sequence has at least 90%, at least 95%, or at least 99% identity to a nucleic acid sequence identified by SEQ ID NO: 16 or 17, but does not include wild-type protein sequence. 
     
     
         21 . The zoonotic disease vaccine of  claim 11 , wherein the RNA comprising an ORF sequence comprises a nucleic acid sequence of SEQ ID NO: 16 or 17. 
     
     
         22 . The zoonotic disease vaccine of  claim 1 , wherein the ORF encodes a middle east respiratory syndrome coronavirus (MERS-CoV) antigen and/or a severe acute respiratory syndrome-like coronavirus WIV1 (SL-CoV-WIV1) antigen. 
     
     
         23 . The zoonotic disease vaccine of  claim 22 , wherein the MERS-CoV antigen and/or a SL-CoV-WIV1 antigen comprises a betacoronavirus structural protein. 
     
     
         24 . The zoonotic disease vaccine of  claim 23 , wherein the betacoronavirus structural protein is spike protein, envelope protein, nucleocapsid protein, or membrane protein. 
     
     
         25 . The zoonotic disease vaccine of  claim 24 , wherein the betacoronavirus structural protein is spike protein. 
     
     
         26 . The zoonotic disease vaccine of  claim 25 , wherein the betacoronavirus structural protein a S1 subunit of the spike protein or a S2 subunit of the spike protein. 
     
     
         27 . The zoonotic disease vaccine of any one of  claims 22 - 26 , wherein the MERS-CoV antigen and/or a SL-CoV-WIV1 antigen is a MERS-CoV antigen. 
     
     
         28 . The zoonotic disease vaccine of any one of  claims 22 - 26 , wherein the MERS-CoV antigen and/or a SL-CoV-WIV1 antigen is a SL-CoV-WIV1 antigen. 
     
     
         29 . The zoonotic disease vaccine of  claim 22 , wherein the MERS-CoV antigen and/or a SL-CoV-WIV1 antigen has an amino acid sequence that has at least 90%, at least 95%, or at least 99% identity to an amino acid sequence identified SEQ ID NO: 18 but does not include wild-type protein sequence. 
     
     
         30 . The zoonotic disease vaccine of  claim 22 , wherein the MERS-CoV antigen and/or a SL-CoV-WIV1 antigen has an amino acid sequence of SEQ ID NO: 18. 
     
     
         31 . The zoonotic disease vaccine of  claim 22 , wherein the RNA comprising an ORF sequence has at least 90%, at least 95%, or at least 99% identity to a nucleic acid sequence identified by SEQ ID NO: 18, but does not include wild-type protein sequence. 
     
     
         32 . The zoonotic disease vaccine of  claim 22 , wherein the RNA comprising an ORF sequence comprises a nucleic acid sequence of SEQ ID NO: 18. 
     
     
         33 . The zoonotic disease vaccine of any one of  claims 1 - 32 , wherein IM administration of a therapeutically effective amount of the vaccine to a subject induces a neutralizing antibody titer in the subject. 
     
     
         34 . The zoonotic disease vaccine of  claim 33 , wherein the neutralizing antibody titer is at least 100 neutralizing units per milliliter (NU/mL), at least 500 NU/mL, or at least 1000 NU/mL. 
     
     
         35 . The zoonotic disease vaccine of  claim 33  or  34 , wherein the neutralizing antibody titer is sufficient to reduce viral infection of B cells by at least 50% relative to a neutralizing antibody titer of an unvaccinated control subject or relative to a neutralizing antibody titer of a subject vaccinated with a live attenuated viral vaccine, an inactivated viral vaccine, or a protein subunit viral vaccine. 
     
     
         36 . The zoonotic disease vaccine of any one of  claims 33 - 35 , wherein the neutralizing antibody titer is induced in the subject following fewer than three doses of the vaccine. 
     
     
         37 . The zoonotic disease vaccine of any one of  claims 1 - 36 , wherein a single dose is of 10 μg-100 μg. 
     
     
         38 . The zoonotic disease vaccine of any one of  claims 33 - 37 , wherein the neutralizing antibody titer and/or a T cell immune response is sufficient to reduce the rate of asymptomatic viral infection relative to the neutralizing antibody titer of unvaccinated control subjects. 
     
     
         39 . The zoonotic disease vaccine of any one of  claims 33 - 38 , wherein the neutralizing antibody titer and/or a T cell immune response is sufficient to prevent viral latency the subject. 
     
     
         40 . The zoonotic disease vaccine of any one of  claims 33 - 39 , wherein the neutralizing antibody titer is sufficient to block fusion of virus with epithelial cells and/or B cells of the subject. 
     
     
         41 . The zoonotic disease vaccine of any one of  claims 33 - 40 , wherein the neutralizing antibody titer is induced within 20 days following a single 10-100 μg of the vaccine, or within 40 days following a second 10-100 μg dose of the vaccine. 
     
     
         42 . The zoonotic disease vaccine of any one of  claims 33 - 40 , wherein IM administration of a therapeutically effective amount of the vaccine to a subject induces a T cell immune response in the subject. 
     
     
         43 . The zoonotic disease vaccine of  claim 42 , wherein the T cell immune response comprises a CD4 +  T cell immune response and/or a CD8 +  T cell immune response. 
     
     
         44 . The zoonotic disease vaccine of any one of  claims 1 - 43 , wherein the antigen is expressed on the surface of cells of the subject. 
     
     
         45 . The zoonotic disease vaccine of any one of  claims 1 - 44 , wherein the vaccine comprises (a) a ribonucleic acid (RNA) having an open reading frame (ORF) encoding two antigens, or (b) two RNAs, each having an ORF encoding an antigen. 
     
     
         46 . The zoonotic disease vaccine of any one of  claims 1 - 45 , wherein the vaccine comprises a RNA having an ORF encoding two antigens formulated in a lipid nanoparticle. 
     
     
         47 . The zoonotic disease vaccine of any one of  claims 1 - 46 , wherein the vaccine comprises two RNAs, each having an ORF encoding an antigen, wherein the two RNAs are formulated in a single lipid nanoparticle or wherein the each RNAs is formulated in a single lipid nanoparticle. 
     
     
         48 . The zoonotic disease vaccine of any one of  claims 1 - 47 , further comprising at least one additional RNA having an ORF encoding at least one additional antigen. 
     
     
         49 . The zoonotic disease vaccine of any one of  claims 46 - 48 , wherein the lipid nanoparticle comprises a molar ratio of 20-60% ionizable cationic lipid, 5-25% non-cationic lipid, 25-55% sterol, and 0.5-15% PEG-modified lipid 
     
     
         50 . The zoonotic disease vaccine of any one of  claims 1 - 49 , wherein the antigen is fused to a signal peptide. 
     
     
         51 . The zoonotic disease vaccine of any one of  claims 1 - 50 , wherein the antigen is fused to a scaffold moiety. 
     
     
         52 . The zoonotic disease vaccine of  claim 51 , wherein the scaffold moiety is selected from the group consisting of: ferritin, encapsulin, lumazine synthase, hepatitis B surface antigen, and hepatitis B core antigen. 
     
     
         53 . The zoonotic disease vaccine of any one of  claims 1 - 52 , wherein the RNA comprises messenger RNA (mRNA). 
     
     
         54 . The zoonotic disease vaccine of any one of  claims 1 - 53 , wherein the RNA further comprises a 5′UTR and/or a 3′UTR. 
     
     
         55 . The zoonotic disease vaccine of any one of  claims 1 - 54 , wherein the RNA is unmodified. 
     
     
         56 . The zoonotic disease vaccine of any one of  claims 1 - 54 , wherein the RNA comprise a modified nucleotide. 
     
     
         57 . The zoonotic disease vaccine of  claim 56 , wherein at least 80% of the uracil in the ORF comprise 1-methyl-pseudouridine modification. 
     
     
         58 . A method comprising administering to a subject the zoonotic disease vaccine of any one of  claims 1 - 57  in a therapeutically effective amount to induce an immune response in the subject. 
     
     
         59 . The method of  claim 58 , wherein the therapeutically effective amount induces a neutralizing antibody titer and/or a T cell immune response in the subject. 
     
     
         60 . The method of  claim 59 , wherein efficacy of the vaccine is at least 80% relative to unvaccinated control subjects. 
     
     
         61 . The method of any one of  claims 58 - 60 , wherein detectable levels of the antigen are produced in the serum of the subject at 1-72 hours post administration of the vaccine. 
     
     
         62 . The method of any one of  claims 59 - 61 , wherein a neutralizing antibody titer of at least 100 NU/ml, at least 500 NU/ml, or at least 1000 NU/ml is produced in the serum of the subject at 1-72 hours post administration of the vaccine. 
     
     
         63 . The method of any one of  claims 58 - 62 , wherein the therapeutically effective amount is a total dose of 20 μg-200 μg or a total dose of 50 μg-100 μg.