Engineered Nipah virus mRNA vaccine

Abstract

Provided herein are (i) messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a soluble Nipah virus glycoprotein (soluble NiV-G) fused to a human type I collagen alpha 1 (COL1A1) signal peptide, (ii) an mRNA comprising an ORF encoding the full-length Nipah virus glycoprotein (full-length NiV-G), (iii) an mRNA comprising an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F), or (iv) an mRNA comprising an ORF encoding full-length NiV-G and an ORF encoding full-length NiV-F, as well as methods for inducing an immune response against Nipah virus by administering an effective amount of the Nipah virus vaccine composition to a subject in need thereof.

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 484,339, filed February 10, 2023, the entire disclosure of which is incorporated herein by reference.

[0002] [Incorporation by reference to sequence listing] The contents of the electronically submitted sequence listing submitted herewith, with file name: F295207_sequence listing as filed, size: 37,709 bytes, and creation date: January 16, 2024, are incorporated herein by reference in their entirety.

[0003] Provided herein are (i) messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a soluble Nipah virus glycoprotein (soluble NiV-G) fused to a human type I collagen alpha 1 (COL1A1) signal peptide, (ii) an mRNA comprising an ORF encoding the full-length Nipah virus glycoprotein (full-length NiV-G), (iii) an mRNA comprising an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F), or (iv) an mRNA comprising an ORF encoding full-length NiV-G and an ORF encoding full-length NiV-F, as well as methods for inducing an immune response against Nipah virus by administering an effective amount of the Nipah virus vaccine composition to a subject in need thereof. [Background technology]

[0004] Nipah virus (NiV) is a zoonotic virus (transmitted from animals to humans) that can be transmitted through contaminated food or directly between humans. Nipah virus causes a range of illnesses in infected individuals, from asymptomatic (subclinical) infection to acute respiratory illness and fatal encephalitis. Currently, there is no approved Nipah virus mRNA vaccine, and a Nipah virus mRNA vaccine is needed. Summary of the Invention

[0005] The present disclosure provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a soluble Nipah virus glycoprotein (soluble NiV-G) fused to the human type I collagen alpha 1 (COL1A1) signal peptide. In one embodiment, the soluble NiV-G fused to the COL1A1 signal peptide has the amino acid sequence of SEQ ID NO: 1. In another embodiment, the ORF encoding the soluble NiV-G fused to the COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the mRNA comprising the ORF encoding soluble NiV-G fused to the COL1A1 signal peptide further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, such that the ORF encoding soluble NiV-G fused to the COL1A1 signal peptide has the structure ORF-3'UTR-poly(A) tail, and the ORF encoding soluble NiV-G fused to the COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO: 2. In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In some embodiments, the mRNA comprising the ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused to the 5'UTR-COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO: 3. In one embodiment, the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused to the 5'UTR-COL1A1 signal peptide has a nucleotide sequence having at least 80% identity to SEQ ID NO: 3. In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0006] The present disclosure also provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a full-length Nipah virus glycoprotein (full-length NiV-G). In one embodiment, the full-length NiV-G has the amino acid sequence of SEQ ID NO: 5. In one embodiment, the ORF encoding full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6. In one embodiment, the mRNA comprising the ORF encoding full-length NiV-G further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, such that the ORF has the structure 5' UTR-ORF encoding full-length NiV-G-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6. In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In one embodiment, the mRNA having the structure 5' UTR-ORF encoding full-length NiV-G-3' UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO: 7. In one embodiment, the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail has a nucleotide sequence having at least 80% identity to SEQ ID NO: 7. In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0007] The present disclosure also provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a full-length Nipah virus fusion protein (full-length NiV-F). In one embodiment, the full-length NiV-F has the amino acid sequence of SEQ ID NO: 9. In one embodiment, the ORF encoding full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10. In one embodiment, the mRNA comprising the ORF encoding full-length NiV-F further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, such that the ORF has the structure 5' UTR-ORF encoding full-length NiV-F-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10. In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In one embodiment, the mRNA having the structure 5' UTR-ORF encoding full-length NiV-F-3' UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO: 11. In one embodiment, the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail has a nucleotide sequence with at least 80% identity to SEQ ID NO: 11. In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0008] The present disclosure also provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding the full-length Nipah virus glycoprotein (full-length NiV-G) and an mRNA comprising an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F). In one embodiment, the full-length NiV-G has the amino acid sequence of SEQ ID NO: 5, and the full-length NiV-F has the amino acid sequence of SEQ ID NO: 9. In one embodiment, the ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6, and the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10. In one embodiment, the mRNA comprising the ORF encoding full-length NiV-G further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has the structure 5' UTR-ORF encoding full-length NiV-G-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6. The mRNA comprising the ORF encoding full-length NiV-F further comprises a 5' UTR, a 3' UTR, and a poly(A) tail, and thus has the structure 5' UTR-ORF encoding full-length NiV-F-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10. In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In one embodiment, the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-G has the nucleotide sequence of SEQ ID NO: 7, and the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-F has the nucleotide sequence of SEQ ID NO: 11. In one embodiment, the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-G has a nucleotide sequence that is at least 80% identical to SEQ ID NO: 7, and the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-F has a nucleotide sequence that is at least 80% identical to SEQ ID NO: 11.In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier, hi one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0009] The present disclosure also provides a method of inducing an immune response against Nipah virus, comprising administering to a subject in need thereof an effective amount of a Nipah virus vaccine composition according to the present disclosure. [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 shows in vitro transcription of NiV mRNA. [Figure 2A-2B] FIG. 1 shows the expression of NiV-G protein and NiV-F protein in mRNA-transfected 293FT cells. [Figure 3A] FIG. 1 illustrates the immunization schedule for mice with the Nipah virus vaccine composition of the present disclosure. [Figure 3B] FIG. 1 shows data on mouse anti-NiV-GP titers. [Figure 3C] FIG. 1 shows data on NiV neutralization. DETAILED DESCRIPTION OF THE INVENTION

[0011] definition Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the embodiments described herein, certain preferred methods, compositions, devices, and materials are described herein. However, before describing the materials and methods of the present invention, it should be understood that this disclosure is not limited to the particular molecules, compositions, methodologies, or protocols described herein, as these may vary depending on routine experimentation and optimization. It should also be understood that the terminology used in the detailed description is for the purpose of describing particular forms or embodiments only, and is not intended to limit the scope of the embodiments described herein.

[0012] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. However, in case of conflict, the present specification, including definitions, will control. Therefore, in the context of the embodiments described herein, the following definitions apply.

[0013] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0014] As used herein, the term "comprise" and its linguistic variants indicate the presence of the recited feature(s), element(s), method step(s), etc., but do not exclude the presence of additional feature(s), element(s), method step(s), etc. In contrast, the term "consisting of" and its linguistic variants indicate the presence of the recited feature(s), element(s), method step(s), etc., and excludes any unrecited feature(s), element(s), method step(s), etc., except for impurities ordinarily associated therewith. The phrase "consisting essentially of" refers to the recited feature(s), element(s), method step(s), etc., as well as any additional feature(s), element(s), method step(s), etc. that do not materially affect the basic characteristics of the composition, system, or method. Many embodiments herein are described using the open-ended term "comprising." Such embodiments encompass a number of the open-ended "consisting of" and / or "consisting essentially of" embodiments, which may alternatively be claimed or described in the specification using such terms.

[0015] As used herein, the term "Nipah virus vaccine composition" refers to substances used to stimulate the production of antibodies and provide immunity against Nipah virus.

[0016] As used herein, the term "messenger ribonucleic acid (mRNA)" refers to a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene and is read by ribosomes in the process of synthesizing proteins.

[0017] As used herein, the term "fused with" refers to a gene or gene product that retains the properties of that gene or gene product itself when isolated from a naturally occurring source.

[0018] The term "Nipah virus glycoprotein fused to human type I collagen alpha 1 (COL1A1) signal peptide (soluble NiV-G)" refers to a recombinant fusion protein created by genetic engineering of a fusion gene. For example, this may require removing the stop codon from the cDNA sequence encoding soluble NiV-G, followed by in-frame addition of the cDNA sequence for the COL1A1 signal peptide through ligation or overlap extension PCR.

[0019] Naturally occurring amino acids include alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N), aspartic acid (Asp or D), cysteine ​​(Cys or C), glutamine (Gln or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine (His or H), isoleucine (Ile or I), leucine (Leu or L), lysine (Lys or K), methionine (Met or M), phenylalanine (Phe or F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), tryptophan (Trp or W), tyrosine (Tyr or Y), and valine (Val or V).

[0020] Unnatural amino acids include azetidine carboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid, β-alanine, naphthylalanine ("naph"), aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, tert-butylglycine ("tBuG"), 2,4-diaminoisobutyric acid, desmosine, 2,2'-diaminopimelic acid, 2,3-diaminopropionic acid, N-ethylglycine, N-ethylasparagine, homoproline ("hPro" or "homoP"), hydroxylysine, allo-hydroxylysine, 3-hydroxyproline ("3Hyp"), 4-hydroxyproline ( "4Hyp"), isodesmosine, allo-isoleucine, N-methylalanine ("MeAla" or "Nime"), N-alkylglycines ("NAG") including N-methylglycine, N-methylisoleucine, N-alkylpentylglycines ("NAPG") including N-methylpentylglycine, N-methylvaline, naphthylalanine, norvaline ("Norval"), norleucine ("Norleu"), octylglycine ("OctG"), ornithine ("Orn"), pentylglycine ("pG" or "PGly"), pipecolic acid, thioproline ("ThioP" or "tPro"), homolysine ("hLys"), and homoarginine ("hArg").

[0021] As used herein, the term "open reading frame (ORF)" refers to the nucleotide sequence between the start codon and the stop codon.

[0022] As used herein, the term "open reading frame (ORF) encoding" refers to a nucleotide coding sequence that encodes a polypeptide. The coding sequence may further include initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of an individual or mammal to which the nucleic acid is administered. The coding sequence may further include a sequence encoding a signal peptide.

[0023] As used herein, the term "T7 promoter" refers to the promoter derived from bacteriophage T7.

[0024] As used herein, the term "5' untranslated region (UTR)" refers to the region of an mRNA immediately upstream (i.e., 5') from the start codon (the first codon of an mRNA transcript that is translated by ribosomes) that does not encode a polypeptide.

[0025] As used herein, the term "3' untranslated region (UTR)" refers to the region of an mRNA immediately downstream (i.e., 3') from the stop codon (i.e., the codon of the mRNA transcript that signals the end of translation) that does not encode a polypeptide.

[0026] As used herein, the term "poly(A) tail" refers to the "tail" of an mRNA, i.e., the long stretch of adenine nucleotides added to the 3' end.

[0027] As used herein, the term "pharmaceutically acceptable carrier" refers to any substance or vehicle suitable for delivering an mRNA vaccine to a suitable in vivo or ex vivo site. Such carriers may include, but are not limited to, adjuvants, excipients, lipid particles, etc.

[0028] As used herein, the term "lipid nanoparticle" refers to a particle having at least one dimension on the nanometer order (e.g., 1 nm to 1000 nm). In some embodiments, lipid nanoparticles are included in formulations that can be used to deliver mRNA vaccines to desired target sites (e.g., cells, tissues, organs, tumors, etc.). In some embodiments, the mRNA vaccine can be encapsulated in the lipid portion of the lipid nanoparticle or in the aqueous space surrounded by some or all of the lipid portion of the lipid nanoparticle, thereby protecting it from enzymatic degradation or other undesirable effects induced by the host organism's or cellular machinery, such as a harmful immune response. In some embodiments, the lipid nanoparticles have an average diameter of 50 nm to 200 nm. In some embodiments, the lipid nanoparticles comprise a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid. In some embodiments, the lipid nanoparticles comprise a molar ratio of approximately 20% to 60% cationic lipid, 0.5% to 15% PEG-modified lipid, 25% to 55% sterol, and 25% non-cationic lipid. In some embodiments, the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is cholesterol. In some embodiments, the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319).

[0029] As used herein, the term "inducing an immune response against Nipah virus" refers to providing protective immunity and / or vaccinating a subject against Nipah virus for prophylactic purposes, and to producing a desired immune response or effect against Nipah virus in a subject in need thereof for therapeutic purposes. As used herein, the term "protective immunity" or "protective immune response" means that a vaccinated subject is able to control infection by the pathogen against which the vaccinated subject is vaccinated. Typically, a subject that develops a "protective immune response" will develop only mild to moderate clinical symptoms, or no symptoms at all.

[0030] An "effective amount" of a Nipah virus vaccine composition (e.g., mRNA) is provided based, at least in part, on the target tissue, target cell type, means of administration, physical characteristics of the polynucleotide (e.g., size and degree of nucleoside modification) and other components of the vaccine, as well as other determinants. Generally, an effective amount of a Nipah virus vaccine (e.g., mRNA) will result in a more efficient immune response than a composition containing a corresponding unmodified polynucleotide encoding the same antigen or peptide antigen, preferably induced or boosted in response to intracellular antigen production. Increased antigen production may be demonstrated by increased cellular transfection (percentage of cells transfected with RNA, e.g., mRNA, vaccine), increased protein translation from the polynucleotide, decreased nucleic acid degradation (e.g., as indicated by an increased duration of protein translation from a modified polynucleotide), or a change in the antigen-specific immune response of the host cell.

[0031] As used herein, the term "X% identity to SEQ ID NO: Y" or "sequence identity" refers to the extent to which two polymer sequences (e.g., peptides, polypeptides, nucleic acids, etc.) have the same sequential composition of monomer subunits. The term "sequence similarity" refers to the extent to which two polymer sequences (e.g., peptides, polypeptides, nucleic acids, etc.) differ only by conservative and / or semi-conservative amino acid substitutions. "Percent sequence identity" (or "percent sequence similarity") is calculated by (1) comparing two optimally aligned sequences over a comparison window (e.g., the length of the longer sequence, the length of the shorter sequence, a specified window, etc.), (2) determining the number of positions containing identical (or similar) monomers (e.g., the same amino acid occurs in both sequences, a similar amino acid occurs in both sequences) to obtain the number of matched positions, (3) dividing the number of matched positions by the total number of positions within the comparison window (e.g., the length of the longer sequence, the length of the shorter sequence, a specified window), and (4) multiplying the result by 100 to obtain the percent sequence identity or percent sequence similarity. For example, if peptide A and peptide B are both 20 amino acids long and have identical amino acids at all positions except one, peptide A and peptide B have 95% sequence identity. If the amino acids at non-identical positions share the same biophysical properties (e.g., both are acidic), peptide A and peptide B will have 100% sequence similarity. As another example, if peptide C is 20 amino acids long and peptide D is 15 amino acids long, and 14 of the 15 amino acids in peptide D are identical to those in a portion of peptide C, peptides C and D will have 70% sequence identity, but peptide D will have 93.3% sequence identity over the optimal comparison window of peptide C. For purposes of calculating "percent sequence identity" (or "percent sequence similarity") herein, any gap in the aligned sequences is treated as a mismatch at that position.

[0032] As used herein, the term "a nucleotide sequence having at least X% identity to sequence number Y and encoding a Z protein" means that the nucleotide sequence satisfies two distinct requirements: having at least X% identity to sequence number Y and encoding a Z protein.

[0033] As used herein, the terms "about," "approximate," "at or about," and "substantially" mean that the relevant amount or value may be the exact value set forth in the claims or taught herein, or a value that will produce an equivalent result or effect. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics are not, and need not be, exact, but may be approximate and / or larger or smaller, if desired, to produce an equivalent result or effect, reflecting tolerances, conversion factors, rounding, measurement errors, and the like, as well as other factors known to those skilled in the art. In some circumstances, it may not be possible to reasonably determine a value that will produce an equivalent result or effect. In such cases, generally, as used herein, "about" and "at or about" are understood to mean a ±10% variation from the nominal value indicated, unless otherwise indicated or implied. In general, amounts, sizes, formulations, parameters, or other quantities or characteristics are "about," "approximate," or "at or about," whether or not expressly stated as such. When "about," "approximate," or "at or about" is used before or after a quantitative value, it is understood that the parameter also includes the particular quantitative value itself, unless otherwise specifically stated.

[0034] The terms "subject," "patient," "individual," and the like are used interchangeably herein and refer to any animal, any mammalian subject, or cells thereof, suitable for the methods described herein, whether in vitro or in situ. In certain non-limiting embodiments, the patient, subject, or individual is a human.

[0035] Detailed Description 1. Nipah virus vaccine composition (1) Nipah virus vaccine composition (1): A Nipah virus vaccine composition comprising an mRNA containing an ORF encoding a soluble Nipah virus glycoprotein (soluble NiV-G) fused with a human type I collagen alpha 1 (COL1A1) signal peptide. The present disclosure provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a soluble Nipah virus glycoprotein (soluble NiV-G) fused to a human type I collagen alpha 1 (COL1A1) signal peptide. In one embodiment, the soluble NiV-G fused to the COL1A1 signal peptide has the amino acid sequence of SEQ ID NO: 1 (or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1). In another embodiment, the ORF encoding the soluble NiV-G fused to the COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO: 2 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 2). In some embodiments, the mRNA comprising the ORF encoding soluble NiV-G fused to the COL1A1 signal peptide further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has the structure: ORF encoding soluble NiV-G fused to 5' UTR-COL1A1 signal peptide-3' UTR-poly(A) tail, and the ORF encoding soluble NiV-G fused to the COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO: 2 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 2). In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In some embodiments, an mRNA having the structure of ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused to a 5'UTR-COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO: 3 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 3).In one embodiment, the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused to the 5'UTR-COL1A1 signal peptide has a nucleotide sequence with at least 80% identity to SEQ ID NO: 3. In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0036] (2) Nipah virus vaccine composition (2): Nipah virus vaccine composition containing mRNA containing an ORF encoding the full-length Nipah virus glycoprotein (full-length NiV-G) The present disclosure also provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a full-length Nipah virus glycoprotein (full-length NiV-G). In one embodiment, the full-length NiV-G has the amino acid sequence of SEQ ID NO: 5 (or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 5). In one embodiment, the ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 6). In one embodiment, the mRNA comprising an ORF encoding full-length NiV-G further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has a structure of 5' UTR-ORF encoding full-length NiV-G-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 6). In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In one embodiment, the mRNA having the structure of 5' UTR-ORF encoding full-length NiV-G-3' UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO: 7 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 7). In one embodiment, the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail has a nucleotide sequence with at least 80% identity to SEQ ID NO: 7. In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0037] (3) Nipah virus vaccine composition (3): Nipah virus vaccine composition containing mRNA containing an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F) The present disclosure also provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding a full-length Nipah virus fusion protein (full-length NiV-F). In one embodiment, the full-length NiV-F has the amino acid sequence of SEQ ID NO:9 (or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:9). In one embodiment, the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO:10 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:10). In one embodiment, the mRNA comprising an ORF encoding full-length NiV-F further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has a structure of 5' UTR-ORF encoding full-length NiV-F-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 10). In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In one embodiment, the mRNA having the structure of 5' UTR-ORF encoding full-length NiV-F-3' UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO: 11 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11). In one embodiment, the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail has a nucleotide sequence having at least 80% identity to SEQ ID NO: 11 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 11). In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0038] (4) Nipah virus vaccine composition (4): Nipah virus vaccine composition containing mRNA containing an ORF encoding the full-length Nipah virus glycoprotein (full-length NiV-G) and an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F) The present disclosure also provides a Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) comprising an open reading frame (ORF) encoding the full-length Nipah virus glycoprotein (full-length NiV-G) and an mRNA comprising an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F). In one embodiment, the full-length NiV-G has the amino acid sequence of SEQ ID NO: 5, and the full-length NiV-F has the amino acid sequence of SEQ ID NO: 9. In one embodiment, the ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6, and the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10. In one embodiment, the mRNA comprising the ORF encoding full-length NiV-G further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has the structure of 5' UTR-ORF encoding full-length NiV-G-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6. The mRNA comprising the ORF encoding full-length NiV-F further comprises a 5' UTR, a 3' UTR, and a poly(A) tail, and thus has the structure of 5' UTR-ORF encoding full-length NiV-F-3' UTR-poly(A) tail, and the ORF encoding full-length NiV-F has the nucleotide sequence of SEQ ID NO: 10. In one embodiment, the poly(A) tail has a length of 50 to 250 nucleotides. In one embodiment, the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-G has the nucleotide sequence of SEQ ID NO: 7, and the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-F has the nucleotide sequence of SEQ ID NO: 11. In one embodiment, the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-G has a nucleotide sequence that is at least 80% identical to SEQ ID NO: 7, and the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full length NiV-F has a nucleotide sequence that is at least 80% identical to SEQ ID NO: 11.In one embodiment, the Nipah virus composition of the present disclosure further comprises a pharmaceutically acceptable carrier. In one embodiment, the mRNA encoding the full-length NiV-G and the mRNA encoding the full-length NiV-F are contained in the composition in a ratio of about 1:1. In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA.

[0039] In the above Nipah virus vaccine compositions (1) to (4), the poly(A) tail may have a length of 50 to 250 nucleotides. In another embodiment, the poly(A) tail has a length of 100 to 200 nucleotides. In another embodiment, the poly(A) tail has a length of 110 to 150 nucleotides. In another embodiment, the poly(A) tail has a length of 115 to 125 nucleotides. In another embodiment, the poly(A) tail has a length of 116 to 124 nucleotides. In another embodiment, the poly(A) tail has a length of 117 to 123 nucleotides. In another embodiment, the poly(A) tail has a length of 118 to 122 nucleotides. In another embodiment, the poly(A) tail has a length of 119 to 122 nucleotides. In another embodiment, the poly(A) tail has a length of 115 nucleotides. In another embodiment, the poly(A) tail has a length of 116 nucleotides. In another embodiment, the poly(A) tail has a length of 117 nucleotides. In another embodiment, the poly(A) tail has a length of 118 nucleotides. In another embodiment, the poly(A) tail has a length of 119 nucleotides. In another embodiment, the poly(A) tail has a length of 120 nucleotides. In another embodiment, the poly(A) tail has a length of 121 nucleotides. In another embodiment, the poly(A) tail has a length of 122 nucleotides. In another embodiment, the poly(A) tail has a length of 123 nucleotides. In another embodiment, the poly(A) tail has a length of 124 nucleotides. In another embodiment, the poly(A) tail has a length of 125 nucleotides.

[0040] In the above-described Nipah virus vaccine compositions (1) to (4), the mRNA of the present disclosure may contain at least one chemical modification selected from the group consisting of pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4'-thiouridine, 5-methylcytosine, 5-methyluridine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine, and 2'-O-methyluridine. In another embodiment, the chemical modification is at the 5-position of uracil. In another embodiment, the chemical modification is N1-methylpseudouridine. In another embodiment, the chemical modification is N1-ethylpseudouridine.

[0041] In the above-described Nipah virus vaccine compositions (1) to (4), the Nipah virus vaccine composition may further comprise a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutically acceptable carrier may comprise any substance or vehicle suitable for delivering the mRNA vaccine to a suitable in vivo or ex vivo site. Such carriers include, but are not limited to, adjuvants, excipients, lipid particles, and the like. The lipid nanoparticles may be particles having at least one dimension on the nanometer order (e.g., 1 nm to 1000 nm). In some embodiments, the lipid nanoparticles are included in a formulation that can be used to deliver the mRNA vaccine to a desired target site (e.g., a cell, tissue, organ, tumor, etc.). In some embodiments, the mRNA vaccine can be encapsulated in the lipid portion of the lipid nanoparticle or in an aqueous space surrounded by some or all of the lipid portion of the lipid nanoparticle, thereby protecting it from enzymatic degradation or other undesirable effects induced by the host organism's or cells' mechanisms, such as a harmful immune response. In some embodiments, the lipid nanoparticles have an average diameter of 50 nm to 200 nm. In some embodiments, the lipid nanoparticles comprise a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid. In some embodiments, the lipid nanoparticles comprise a molar ratio of about 20%-60% cationic lipid, 0.5%-15% PEG-modified lipid, 25%-55% sterol, and 25% non-cationic lipid. In some embodiments, the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is cholesterol. In some embodiments, the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319).

[0042] In one embodiment, the lipid nanoparticles comprise (i) at least one lipid selected from the group consisting of 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319); (ii) a neutral lipid selected from DSPC, DPPC, POPC, DOPE, and SM; (iii) a sterol, such as cholesterol; and (iv) a PEG lipid, such as PEG-DMG or PEG-cDMA, in a molar ratio of about 20%-60% cationic lipid:5%-25% neutral lipid:25%-55% sterol:0.5%-15% PEG lipid.

[0043] In one embodiment, the lipid nanoparticles comprise about 25% to about 75% on a molar basis, e.g., about 35% to about 65%, about 45% to about 65%, about 60%, about 57.5%, about 50%, or about 40% on a molar basis, of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319).

[0044] In one embodiment, the lipid nanoparticles contain about 0.5% to about 15% neutral lipid on a molar basis, e.g., about 3% to about 12%, about 5% to about 10%, or about 15%, about 10%, or about 7.5%. Examples of neutral lipids include, but are not limited to, DSPC, POPC, DPPC, DOPE, and SM. In some embodiments, the formulation contains about 5% to about 50% sterol on a molar basis (e.g., about 15% to about 45%, about 20% to about 40%, about 40%, about 38.5%, about 35%, or about 31%). An exemplary sterol is cholesterol. In some embodiments, the formulation comprises about 0.5% to about 20% PEG-lipid or PEG-modified lipid on a molar basis (e.g., about 0.5% to about 10%, about 0.5% to about 5%, about 1.5%, about 0.5%, about 1.5%, about 3.5%, or about 5% on a molar basis). In some embodiments, the PEG-lipid or PEG-modified lipid comprises PEG molecules with an average molecular weight of 2000 Da. In other embodiments, the PEG-lipid or PEG-modified lipid comprises PEG molecules with an average molecular weight of less than 2000, e.g., around 1500 Da, around 1000 Da, or around 500 Da. Examples of PEG-modified lipids include, but are not limited to, PEG-distearoylglycerol (PEG-DMG) (also referred to herein as PEG-C14 or C14-PEG) and PEG-cDMA.

[0045] In one embodiment, the lipid nanoparticles comprise, on a molar basis, 25% to 75% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 0.5% to 15% of a neutral lipid, 5% to 50% of a sterol, and 0.5% to 20% of a PEG lipid or a PEG-modified lipid.

[0046] In one embodiment, the lipid nanoparticles comprise, on a molar basis, 35% to 65% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 3% to 12% of a neutral lipid, 15% to 45% of a sterol, and 0.5% to 10% of a PEG lipid or a PEG-modified lipid.

[0047] In one embodiment, the lipid nanoparticles comprise, on a molar basis, 45% to 65% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 5% to 10% of a neutral lipid, 25% to 40% of a sterol, and 0.5% to 10% of a PEG lipid or a PEG-modified lipid.

[0048] In one embodiment, the lipid nanoparticles comprise about 60% by molar of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 7.5% neutral lipid, about 31% sterol, and about 1.5% PEG lipid or PEG-modified lipid.

[0049] In one embodiment, the lipid nanoparticles comprise about 50% by molar of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 10% neutral lipid, about 38.5% sterol, and about 1.5% PEG lipid or PEG-modified lipid.

[0050] In one embodiment, the lipid nanoparticles comprise about 50% by molar cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 10% neutral lipid, about 35% sterol, about 4.5% or about 5% PEG lipid or PEG-modified lipid, and about 0.5% targeting lipid.

[0051] In one embodiment, the lipid nanoparticles comprise about 40% by molar of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 15% neutral lipid, about 40% sterol, and about 5% PEG lipid or PEG-modified lipid.

[0052] In one embodiment, the Nipah virus vaccine composition of the present disclosure can be delivered, localized, and / or concentrated to a specific location using the delivery methods described below. As a non-limiting example, empty polymer particles can be administered to a subject before, simultaneously with, or after delivery of the Nipah virus vaccine composition of the present disclosure to the subject. Upon contact with the subject, the empty polymer particles undergo a volume change and become lodged, embedded, immobilized, or trapped at a specific location within the subject.

[0053] In another embodiment, the Nipah virus vaccine composition of the present disclosure can be formulated in an active agent release system. For example, the active agent release system can include at least one nanoparticle bound to an oligonucleotide inhibitor chain hybridized with a catalytically active nucleic acid and a compound bound to at least one substrate molecule that is bound to a therapeutically active agent (e.g., a polynucleotide described herein), wherein the therapeutically active agent is released upon cleavage of the substrate molecule by the catalytically active nucleic acid.

[0054] In another embodiment, the Nipah virus vaccine composition of the present disclosure may be formulated into nanoparticles comprising an inner core comprising non-cellular material and an outer surface comprising a cellular membrane, which may be derived from a cell or a virus.

[0055] In another embodiment, the Nipah virus vaccine composition of the present disclosure may be formulated in porous nanoparticle-supported lipid bilayers (protocells).

[0056] In another embodiment, the Nipah virus vaccine compositions of the present disclosure may be formulated into polymeric nanoparticles that have a high glass transition temperature.

[0057] In another embodiment, the Nipah virus vaccine composition of the present disclosure can be formulated into nanoparticles for use in imaging. As a non-limiting example, a liposome can comprise gadolinium(III) 2-{4,7-bis-carboxymethyl-10-[(N,N-distearylamidomethyl-N'-amido-methyl]-1,4,7,10-tetra-azacyclododec-1-yl}-acetate and a neutral, fully saturated phospholipid component.

[0058] The nanoparticles of the present disclosure may further comprise nutrients, such as, but not limited to, nutrients whose deficiency can cause adverse health effects ranging from anemia to neural tube defects. By way of non-limiting example, the nutrient can be iron in the form of ferrous salts, ferric salts, or elemental iron, iodine, folic acid, vitamins, or micronutrients.

[0059] In another embodiment, the Nipah virus vaccine composition of the present disclosure may be formulated into swellable nanoparticles.

[0060] In another embodiment, the Nipah virus vaccine composition of the present disclosure may be formulated in polyanhydride nanoparticles.

[0061] The nanoparticles and microparticles of the present disclosure can be geometrically engineered to modulate macrophage and / or immune responses. In some embodiments, the geometrically engineered particles can have various shapes, sizes, and / or surface charges to incorporate polynucleotides of the present disclosure for targeted delivery, such as, but not limited to, pulmonary delivery. Other physical features that the geometrically engineered particles can have include, but are not limited to, fenestrations, angled arms, asymmetry and surface roughness, and charge, which can alter interactions with cells and tissues.

[0062] In another embodiment, the nanoparticles of the present disclosure may be water-soluble nanoparticles. The nanoparticles may be inorganic nanoparticles that are compact and have zwitterionic ligands to exhibit good water solubility. The nanoparticles may also have a small hydrodynamic diameter (HD), stability over time, pH, and salt concentration, and low levels of nonspecific protein binding.

[0063] In some embodiments, the nanoparticles of the present disclosure are stealth nanoparticles or target-specific stealth nanoparticles. In some embodiments, the stealth nanoparticles or target-specific stealth nanoparticles can comprise a polymer matrix. The polymer matrix can comprise two or more polymers, such as, but not limited to, polyethylene, polycarbonate, polyanhydride, polyhydroxy acid, polypropyl fumarate, polycaprolactone, polyamide, polyacetal, polyether, polyester, poly(orthoester), polycyanoacrylate, polyvinyl alcohol, polyurethane, polyphosphazene, polyacrylate, polymethacrylate, polycyanoacrylate, polyurea, polystyrene, polyamine, polyester, polyanhydride, polyether, polyurethane, polymethacrylate, polyacrylate, polycyanoacrylate, or a combination thereof.

[0064] In one embodiment, the nanoparticles of the present disclosure may be nanoparticle-nucleic acid hybrid structures having a high-density nucleic acid layer. The nanoparticles of the present disclosure may comprise nucleic acids, such as, but not limited to, polynucleotides described herein and / or known in the art.

[0065] In one embodiment, at least one of the nanoparticles of the present disclosure may be embedded in the core of the nanostructure or coated with a low-density porous 3D structure or coating capable of carrying or associating at least one payload within or on the surface of the nanostructure.

[0066] In one embodiment, the pharmaceutically acceptable carrier is a lipid nanoparticle encapsulating the mRNA of the present disclosure. In another embodiment, the lipid nanoparticle comprises a first lipid nanoparticle encapsulating an mRNA encoding soluble NiV-G fused with a COL1A1 signal peptide, a second lipid nanoparticle encapsulating an mRNA encoding full-length NiV-G, a third lipid nanoparticle encapsulating an mRNA encoding full-length NiV-F, and a fourth lipid nanoparticle encapsulating an mRNA encoding full-length NiV-G and an mRNA encoding full-length NiV-F.

[0067] 2. Methods for inducing immune responses against Nipah virus The present disclosure also provides a method for inducing an immune response against Nipah virus, comprising administering an effective amount of a Nipah virus vaccine composition of the present disclosure to a subject in need thereof. In one embodiment, an effective amount of a Nipah virus vaccine composition (e.g., mRNA) is provided based, at least in part, on the target tissue, target cell type, means of administration, physical characteristics of the polynucleotide (e.g., size and degree of modified nucleosides) and other components of the vaccine, as well as other determinants. Generally, an effective amount of a Nipah virus vaccine (e.g., mRNA) induces or boosts an immune response in response to intracellular antigen production, preferably more efficiently than a composition containing a corresponding unmodified polynucleotide encoding the same antigen or peptide antigen. Increased antigen production may be demonstrated by increased cell transfection (percentage of cells transfected with RNA, e.g., mRNA, vaccine), increased protein translation from the polynucleotide, decreased nucleic acid degradation (e.g., indicated by an increased duration of protein translation from a modified polynucleotide), or a change in the antigen-specific immune response of the host cell.

[0068] An effective (immunogenically effective) amount of a Nipah virus vaccine composition (e.g., Nipah virus vaccine compositions (1) to (4)) is typically administered intramuscularly or subcutaneously. Therefore, Nipah virus vaccine compositions are typically formulated for intramuscular or subcutaneous injection, and for purposes of the present invention, they are formulated without an adjuvant, preferably without any adjuvant. However, other modes of administration, such as intravenous, cutaneous, intradermal, or intranasal, are also contemplated. For intravenous, cutaneous, or subcutaneous injection, the adenoviral vector is in the form of a parenterally acceptable aqueous solution that is pyrogen-free and has suitable pH, isotonicity, and stability. Similarly, the isolated envelope polypeptide is in the form of a parenterally acceptable solution that has suitable pH, isotonicity, and stability. Those skilled in the art can prepare suitable solutions using isotonic vehicles, such as sodium chloride injection, Ringer's injection, or lactated Ringer's injection. Preservatives, stabilizers, buffers, antioxidants, and / or other additives may be included as needed.

[0069] In certain embodiments, an effective (immunogenically effective) amount of a Nipah virus vaccine composition (e.g., Nipah virus vaccine compositions (1)-(4)) is administered intramuscularly. Intramuscular administration can be achieved by injecting a suspension of the adenoviral vector and / or envelope polypeptide using a needle. Alternatively, a needleless injection device is used to administer the composition (e.g., using a Biojector™) or a lyophilized powder containing the vaccine.

[0070] In one embodiment, the priming immunization and / or the boosting administration, preferably both the priming administration and the boosting administration, further comprises administering one or more adenoviral vectors encoding one or more additional Nipah virus antigens.

[0071] The timing of priming and boosting immunizations is not particularly limited. For example, the vaccine composition can be administered for priming immunization and then re-administered before administering the vaccine composition for boosting immunization. Further administration of the vaccine composition for further boosting immunization is also contemplated. In certain embodiments, the booster vaccine is first administered about 1 to 12 weeks after the initial administration of the primer vaccine, for example, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks later. In another embodiment, the booster vaccine is first administered about 12 to 52 weeks after the initial administration of the primer vaccine, for example, about 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, or 52 weeks later. Those skilled in the art can modify the exact timing, frequency of administration, dosage, etc. of the priming and boosting vaccines based on the teachings herein and general knowledge in the art.

[0072] In one embodiment, a Nipah virus vaccine composition may be formulated with lipid nanoparticles containing MC3, cholesterol, DSPC, and PEG2000-DMG, a buffer of trisodium citrate, sucrose, and water for injection, comprising the first and second mRNAs described herein. As a non-limiting example, the composition may contain 2.0 mg / mL of the active ingredient (e.g., Nipah virus vaccine compositions (1) to (4)), 21.8 mg / mL of MC3, 10.1 mg / mL of cholesterol, 5.4 mg / mL of DSPC, 2.7 mg / mL of PEG2000-DMG, 5.16 mg / mL of trisodium citrate, 71 mg / mL of sucrose, and 1.0 mL of water for injection.

[0073] In one embodiment, a method for inducing an immune response against Nipah virus comprises administering an effective amount of the Nipah virus vaccine composition (1) of the present disclosure to a subject in need thereof. In the Nipah virus vaccine composition (1), the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused to the 5'UTR-COL1A1 signal peptide may have the nucleotide sequence of SEQ ID NO: 3 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99% identity to SEQ ID NO: 3).

[0074] In another embodiment, a method for inducing an immune response against Nipah virus comprises administering an effective amount of the Nipah virus vaccine composition (2) of the present disclosure to a subject in need thereof. In the Nipah virus vaccine composition (2), the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail may have the nucleotide sequence of SEQ ID NO: 7 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99% identity to SEQ ID NO: 7).

[0075] In another embodiment, a method for inducing an immune response against Nipah virus comprises administering an effective amount of the Nipah virus vaccine composition (3) of the present disclosure to a subject in need thereof. In the Nipah virus vaccine composition (3), the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail may have the nucleotide sequence of SEQ ID NO: 11 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99% identity to SEQ ID NO: 11).

[0076] In another embodiment, a method for inducing an immune response against Nipah virus comprises administering an effective amount of the Nipah virus vaccine composition (4) of the present disclosure to a subject in need thereof. In the Nipah virus vaccine composition (4), the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full-length NiV-G may have the nucleotide sequence of SEQ ID NO: 7 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99% identity to SEQ ID NO: 7), and the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding 5'UTR-full-length NiV-F may have the nucleotide sequence of SEQ ID NO: 11 (or a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99% identity to SEQ ID NO: 11).

[0077] 3. Sequence information 1) Protein sequence of soluble NiV-G fused to the COL1A1 signal peptide (COL1A1 signal peptide sequence is underlined). SEQ ID NO: 1 TIFF2026505581000002.tif48170

[0078] 2) mRNA sequence (ORF) of soluble NiV-G fused with the COL1A1 signal peptide SEQ ID NO: 2

[0079] 3) mRNA sequence of soluble NiV-G fused to the COL1A1 signal peptide (5'UTR-ORF-3'UTR-poly(A) tail) (ORF is underlined). SEQ ID NO: 3 TIFF2026505581000003.tif170170

[0080] 4) Sequence of pUC57-Kan plasmid encoding soluble NiV-G mRNA fused to the COL1A1 signal peptide (the soluble NiV-G mRNA sequence fused to the COL1A1 signal peptide is underlined). SEQ ID NO:4 TIFF2026505581000004.tif157170TIFF2026505581000005.tif228170

[0081] 5) Full-length NiV-G protein sequence SEQ ID NO:5 MPTESKKVRFENTASDKGKNPSKVIKSYYGTMDIKKINEGLLDSKILSAFNTVIALLGSIVIIVMNIMIIQNYTRSTDNQAMIKDALQSIQQQIKGLADKIGTEIGPKVSLIDTSSTITIPANIGLLGSKISQSTASINENVNEKCKFTLPPLKIHECNISCPNPLPFREYKPQTEGVSNLVGLPNNICLQKTSNQILKPKLISYTLPVVGQSGTCITDPLLAMDEGYFAYSHLEKIGSCSRGVSKQRIIGVGEVLDRGDEVPSLFMTNVWTPSNPNTVYHCSAVYNSEFYYVLCAVSVVGDPILNSTYWSGSLMMTRLAVKPKNNGESYNQHQFALRNIEKGMYDKVMPYGPSGIKQGDTLYFPAVGFLVRTEFKYNDSNCPVAKCQYSKPENCRLSMGIRPNSHYILRSGLLKYNLSDEENSKIIFIEISDQRLSIGSPSKIYDSLGQPVFYQASFSWDTMIKFGDVQTVNPLVVNWRDNTVISRPGQSQCPRFNTCPEVCWEGVYNDAFLIDRINWISAGVFLDSNQTAENPVFTVFKDNEVLYRAQLASEDTNAQKTITNCFLLKNKIWCISLVEIYDTGDNVIRPKLFAVKIPEQCT

[0082] 6) Full-length NiV-G mRNA sequence (ORF) SEQ ID NO: 6

[0083] 7) Full-length NiV-G mRNA sequence (5'UTR-ORF-3'UTR-poly(A) tail) (ORFs are underlined). SEQ ID NO:7 TIFF2026505581000006.tif184170

[0084] 8) Sequence of pUC57-Kan plasmid encoding full-length NiV-G (the full-length NiV-G mRNA sequence is underlined). SEQ ID NO:8 TIFF2026505581000007.tif190170TIFF2026505581000008.tif203170

[0085] 9) Full-length NiV-F protein sequence SEQ ID NO:9 MAVILNKRYYSNLLLLILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYE AMKNADNINNKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSG YYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTL LMIDNTTCPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDKVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLYYIGT

[0086] 10) Full-length NiV-F mRNA sequence (ORF) SEQ ID NO: 10

[0087] 11) Full-length NiV-F mRNA sequence (5'UTR-ORF-3'UTR-poly(A) tail) (ORFs are underlined). SEQ ID NO: 11 TIFF2026505581000009.tif170170

[0088] 12) Sequence of pUC57-Kan plasmid encoding full-length NiV-F (the full-length NiV-F mRNA sequence is underlined). SEQ ID NO: 12 TIFF2026505581000010.tif246170TIFF2026505581000011.tif137170 [Example]

[0089] Example 1 - DNA templates used for in vitro transcription and protein expression The DNA template sequence for mRNA in vitro transcription (IVT) consisted of a T7 promoter, a 5' untranslated region (UTR), the open reading frame (ORF) of the Nipah virus glycoprotein (NiV-G) and fusion protein (NiV-F) modified from the MCL-18-H-1088 strain (GenBank: MH523642.1), a 3' UTR, and 120 bases of polyadenine (polyA). The 5' UTR and 3' UTR were derived from the human hemoglobin subunit α1 (HBA1) mRNA (GenBank: NM_000558.5). For the soluble form of the glycoprotein (NiV-G_sol), the amino-terminus and transmembrane domain were removed from the full-length glycoprotein sequence to obtain a potential secretory form (ΔMet1-Ile70). The signal peptide from collagen alpha 1 (COL1A1) (ColSP, MFSFVDLRLLLLLAATALLTHG, GenBank: Z74615.1) was added to the N-terminus of the ORFs of the full-length and soluble glycoproteins to promote their secretion into the culture medium. The initial NiV-G DNA was synthesized by GenScript (Piscataway, NJ) and subcloned into the pUC57-Kan vector. Subsequently, NiV-G_sol, ColSP-NiV-G, and ColSP-NiV-G_sol constructs were generated through PCR and subcloning into the NiV-G construct. The sequences were verified by Sanger sequencing (Azenta, Inc.). NiV-F DNA was synthesized by Twist BioScience and subsequently subcloned into the pUC57-Kan vector. Along with the NiV-F constructs containing ColSP (ColSP-NiV-F) and not containing ColSP (NiV-F), FLAG-tagged constructs (ColSP-NiV-F-FLAG and NiV-F-FLAG) were generated via PCR to ensure protein expression and detection.

[0090] The sequence of the pUC57-Kan plasmid encoding the soluble NiV-G mRNA fused to the COL1A1 signal peptide is shown in SEQ ID NO: 4. The sequence of the pUC57-Kan plasmid encoding the full-length NiV-G mRNA is shown in SEQ ID NO: 8. The sequence of the pUC57-Kan plasmid encoding the full-length NiV-F mRNA is shown in SEQ ID NO: 12.

[0091] Example 2 - In vitro transcription (IVT) For each NiV form, the plasmid vector was linearized with the restriction enzyme BspQI (New England Biolabs). N1-methylpseudouridine (m1Ψ) was purchased from BOC Sciences (Shirley, NY). IVT conditions followed the manufacturer's recommendations (TranscriptAid T7 High Yield Transcription Kit, ThermoFisher) as follows: ATP / CTP / GTP / m1ψTP: 5mM each SmartCap (SC101, ST Pharm): 4mM Linear template DNA: 1 μg of plasmid or 0.5 μg of PCR product T7 RNA polymerase enzyme mix: 2 μl

[0092] IVT was performed in 20 μl reactions by incubation at 37°C for 2 hours. Template DNA was removed with 2 units of DNase I (Invitrogen) for 15 minutes at 37°C, followed by column purification (Monarch RNA Cleanup Kit, New England Biolabs).

[0093] Agarose gel analysis of the IVT products of four NiV-G constructs (A) and four NiV-F constructs (B) revealed mRNAs approximately 2000 nucleotides long for these four mRNAs. After IVT from the DNA templates of NiV-G mRNA (A) and NiV-F mRNA (B), 100 ng of mRNA was run on 1% E-GEL EX agarose in an E-Gel Power Snap electrophoresis device (ThermoFisher) (one of three independent IVT products). See Figure 1.

[0094] Example 3 - Transfection 1 μg of mRNA (synthesized in triplicate) was individually transfected into 293FT cells (Invitrogen) in 12-well plates using 2 μl of Lipofectamine MesseangerMax (Invitrogen) at a 1:2 ratio according to the manufacturer's protocol. 24 hours after transfection, samples were collected from both the medium and cells. Cell lysates were prepared in NP-40 lysis buffer (150 mM sodium chloride / 1% NP-40 / 50 mM Tris (pH 8.0)). 0.1 μg of EGFP mRNA (L-7601, TriLink) was co-transfected as a transfection control.

[0095] Example 4 - Western Blot Rabbit anti-Nipah virus glycoprotein antibody (NIV11-S) was purchased from Alpha Diagnostics (San Antonio, TX). Protein detection was performed using an HRP-conjugated secondary antibody (Jackson ImmunoResearch, West Grove, PA) and SuperSignal West Pico Plus chemiluminescent substrate (Thermo Scientific). GAPDH was detected as a loading control with an HRP-conjugated mouse monoclonal antibody (sc-47724, Santa Cruz Biotechnology). EGFP, a transfection control, was detected with a mouse monoclonal antibody (sc-9996, Santa Cruz Biotechnology). NiV-F protein detection was performed using an anti-FLAG tag monoclonal antibody (GenScript) and an anti-Nipah virus F F1 rabbit antibody (Absolute Antibody, Wilton, UK).

[0096] As shown in Figure 2A, NiV-G protein levels were determined by Western blot. 293FT cells were individually transfected with 1 μg of the four NiV-G mRNAs. Both cell lysates and culture medium were collected 24 h posttransfection and subjected to Western blot analysis using NiV-G-specific antibodies. GFP mRNA was cotransfected as an mRNA transfection control, and untransfected 293FT cells were used as a negative control. GAPDH was used as a loading control. As shown in Figure 2B, NiV-F protein with or without ColSP was detected in cell lysates with FLAG tag and NiV-F-specific antibodies, revealing the 60 kDa full-length NiV-F protein (arrow) and smaller truncated proteins. Untransfected cell lysates and medium were used as negative controls ((-) control).

[0097] Western blots were performed on both samples collected from the culture medium to detect any secreted NiV-G protein and samples collected from the cell lysate to detect intracellular / non-secreted NiV-G protein. As shown in Figure 2A, wild-type NiV-G protein without the COL1A1 signal peptide (lane 2) and soluble NiV-G with the COL1A1 signal peptide (lane 5) were detected in the cell lysate. However, only soluble NiV-G protein with the COL1A1 signal peptide was detected in the culture medium. NiV-G and ColSP-NiV-G_sol were selected as our mRNA vaccine candidates because they were more fully expressed than ColSP-NiV-G and NiV-G_sol. For NiV-F protein, cell lysates were probed with FLAG antibody and further confirmed with a NiV-F-specific antibody (Figure 2B). Similar to NiV-G, wild-type NiV-F was expressed better than ColSP-NiV-F in its full-length form, and therefore wild-type full-length NiV-F was selected as another mRNA vaccine candidate.

[0098] Example 5 - Immunogenicity Studies This study was designed to test the immunogenicity in mice of the Nipah virus vaccine compositions of the present disclosure (e.g., Nipah virus vaccine compositions (1) and (4)).

[0099] Mice were immunized intramuscularly (IM) with the Nipah virus vaccine composition of the present disclosure, either chemically modified or unmodified. Two immunizations were administered, three weeks apart (i.e., at week 0 and week 3), and serum was collected after each immunization. See Figure 3A. Serum antibody titers against soluble NiV-G fused to the COL1A1 signal peptide and full-length NiV-G were determined using a mouse anti-Nipah virus glycoprotein IgG ELISA kit (NIV-025, Alpha Diagnostics). Overall, both mRNA vaccine formulations (VER-012 (Nipah virus vaccine composition (1)) containing soluble NiV-G and VER-015 (Nipah virus vaccine composition (4)) containing full-length NiV-G and NiV-F) demonstrated high titers of IgG antibodies against NiV-G in a dose-dependent manner, as shown in Figure 3B. In addition, serum collected from each mouse was used to perform an in vitro protection assay against Nipah virus, as shown in Figure 3C. Both vaccine formulations demonstrated potent neutralizing activity against Nipah virus in a dose-dependent manner. Overall, the full-length formulation of NiV-G and NiV-F (VER-015) performed better in neutralizing Nipah virus than the soluble NiV-G (VER-012).

[0100] Example 6 - Additional immunogenicity studies This study is designed to test the immunogenicity in mice of the Nipah virus vaccine compositions of the present disclosure (e.g., Nipah virus vaccine compositions (2) and (3)).

[0101] Mice are immunized intramuscularly (IM) with a Nipah virus vaccine composition of the present disclosure, which may or may not be chemically modified. A total of two immunizations are administered, three weeks apart (i.e., at week 0 and week 3), with serum collected after each immunization.

[0102] Both mRNA vaccine formulations (VER-013 (Nipah virus vaccine composition (2)) containing full-length NiV-G and VER-014 (Nipah virus vaccine composition (3)) containing full-length NiV-F) are expected to demonstrate high titers of IgG antibodies against Nipah virus in a dose-dependent manner. In addition, serum collected from each mouse will be used to perform an in vitro NiV neutralization assay against Nipah virus. Both vaccine formulations are expected to demonstrate potent neutralizing activity against Nipah virus.

Claims

1. A Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) containing an open reading frame (ORF) encoding a soluble Nipah virus glycoprotein (soluble NiV-G) fused to a human type I collagen alpha 1 (COL1A1) signal peptide.

2. 2. The Nipah virus vaccine composition of claim 1, wherein the soluble NiV-G fused with the COL1A1 signal peptide has the amino acid sequence of SEQ ID NO:

1.

3. 2. The Nipah virus vaccine composition of claim 1, wherein the ORF encoding the soluble NiV-G fused to the COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO:

2.

4. The mRNA comprising the ORF encoding soluble NiV-G fused to the COL1A1 signal peptide further comprises a 5' untranslated region (UTR), a 3' UTR and a poly(A) tail, thus having the following structure: ORF-3′UTR-poly(A) tail encoding soluble NiV-G fused to 5′UTR-COL1A1 signal peptide; and 2. The Nipah virus vaccine composition of claim 1, wherein the ORF encoding the soluble NiV-G fused to the COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO:

2.

5. 5. The Nipah virus vaccine composition of claim 4, wherein the poly(A) tail has a length of 50 to 250 nucleotides.

6. The Nipah virus vaccine composition of claim 4, wherein the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused with the 5'UTR-COL1A1 signal peptide has the nucleotide sequence of SEQ ID NO:

3.

7. The Nipah virus vaccine composition of claim 4, wherein the mRNA having the structure of ORF-3'UTR-poly(A) tail encoding soluble NiV-G fused with the 5'UTR-COL1A1 signal peptide has a nucleotide sequence having at least 80% identity to SEQ ID NO:

3.

8. 10. A method of inducing an immune response against Nipah virus, comprising administering an effective amount of the Nipah virus vaccine composition of claim 1 to a subject in need thereof.

9. A Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) containing an open reading frame (ORF) encoding the full-length Nipah virus glycoprotein (full-length NiV-G).

10. 10. The Nipah virus vaccine composition of claim 9, wherein the full-length NiV-G has the amino acid sequence of SEQ ID NO:

5.

11. 10. The Nipah virus vaccine composition of claim 9, wherein the ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO:

6.

12. The mRNA containing the ORF encoding the full-length NiV-G further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has the following structure: 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail; and 10. The Nipah virus vaccine composition of claim 9, wherein the ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO:

6.

13. 13. The Nipah virus vaccine composition of claim 12, wherein the poly(A) tail has a length of 50 to 250 nucleotides.

14. The Nipah virus vaccine composition according to claim 12, wherein the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO:

7.

15. The Nipah virus vaccine composition of claim 12, wherein the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail has a nucleotide sequence having at least 80% identity to SEQ ID NO:

7.

16. 10. A method of inducing an immune response against Nipah virus, comprising administering an effective amount of the Nipah virus vaccine composition of claim 9 to a subject in need thereof.

17. A Nipah virus vaccine composition comprising a messenger ribonucleic acid (mRNA) containing an open reading frame (ORF) encoding a full-length Nipah virus fusion protein (full-length NiV-F).

18. 18. The Nipah virus vaccine composition of claim 17, wherein the full-length NiV-F has the amino acid sequence of SEQ ID NO:

9.

19. 18. The Nipah virus vaccine composition of claim 17, wherein the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO:

10.

20. The mRNA containing the ORF encoding the full-length NiV-F further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has the following structure: 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail; and 18. The Nipah virus vaccine composition of claim 17, wherein the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO:

10.

21. 21. The Nipah virus vaccine composition of claim 20, wherein the poly(A) tail has a length of 50 to 250 nucleotides.

22. The Nipah virus vaccine composition of claim 20, wherein the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO:

11.

23. The Nipah virus vaccine composition of claim 20, wherein the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail has a nucleotide sequence having at least 80% identity to SEQ ID NO:

11.

24. 20. A method of inducing an immune response against Nipah virus, comprising administering an effective amount of the Nipah virus vaccine composition of claim 17 to a subject in need thereof.

25. a messenger ribonucleic acid (mRNA) containing an open reading frame (ORF) encoding the full-length Nipah virus glycoprotein (full-length NiV-G); an mRNA containing an ORF encoding the full-length Nipah virus fusion protein (full-length NiV-F); 1. A Nipah virus vaccine composition comprising:

26. The full-length NiV-G has the amino acid sequence of SEQ ID NO: 5, and 26. The Nipah virus vaccine composition of claim 25, wherein the full-length NiV-F has the amino acid sequence of SEQ ID NO:

9.

27. The ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6, and 26. The Nipah virus vaccine composition of claim 25, wherein the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO:

10.

28. The mRNA containing the ORF encoding the full-length NiV-G further comprises a 5' untranslated region (UTR), a 3' UTR, and a poly(A) tail, and thus has the following structure: 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail; and The ORF encoding the full-length NiV-G has the nucleotide sequence of SEQ ID NO: 6, and The mRNA containing the ORF encoding the full-length NiV-F further comprises a 5′ UTR, a 3′ UTR, and a poly(A) tail, and thus has the following structure: 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail; and 26. The Nipah virus vaccine composition of claim 25, wherein the ORF encoding the full-length NiV-F has the nucleotide sequence of SEQ ID NO:

10.

29. 29. The Nipah virus vaccine composition of claim 28, wherein the poly(A) tail has a length of 50 to 250 nucleotides.

30. The mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO: 7, and The Nipah virus vaccine composition of claim 28, wherein the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail has the nucleotide sequence of SEQ ID NO:

11.

31. The mRNA having the structure of 5'UTR-ORF encoding full-length NiV-G-3'UTR-poly(A) tail has a nucleotide sequence having at least 80% identity to SEQ ID NO: 7, and The Nipah virus vaccine composition of claim 28, wherein the mRNA having the structure of 5'UTR-ORF encoding full-length NiV-F-3'UTR-poly(A) tail has a nucleotide sequence having at least 80% identity to SEQ ID NO:

11.

32. 26. A method of inducing an immune response against Nipah virus, comprising administering an effective amount of the Nipah virus vaccine composition of claim 25 to a subject in need thereof.

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