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EP-4735032-A2 - COMPOSITIONS FOR PREVENTION OF CARDIOMYOPATHY SYNDROME

EP4735032A2EP 4735032 A2EP4735032 A2EP 4735032A2EP-4735032-A2

Abstract

The present disclosure provides exemplary sequences and compositions that can be used to active immunization of animals to aid in the prevention of cardiomyopathy syndrome (CMS) caused by Piscine Myocarditis Virus (PMCV). Vaccines and kits comprising the sequences and compositions are also provided, as well as methods of administering the vaccine to non-human animals.

Inventors

  • MACDONALD, Alicia
  • SNYDER, JONATHAN
  • EVENSEN, Øystein

Assignees

  • Intervet International B.V.

Dates

Publication Date
20260506
Application Date
20240626

Claims (20)

  1. 1. An expression vector encoding a piscine myocarditis virus (PMCV) polyprotein.
  2. 2. The expression vector of claim 1, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence selected from a group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
  3. 3. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.
  4. 4. The expression vector of claim 1, wherein the PMCV polyprotein comprises SEQ ID NO: 1.
  5. 5. The expression vector of claim 1, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 1.
  6. 6. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 1.
  7. 7. The expression vector of claim 1, wherein the PMCV polyprotein comprises SEQ ID NO: 2.
  8. 8. The expression vector of claim 1, wherein the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2.
  9. 9. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 2.
  10. 10. The expression vector of claim 1 , wherein the PMCV polyprotein comprises SEQ ID NO: 3.
  11. 11. The expression vector of claim 1, wherein the PMCV polyprote in is at least 90% identical with an amino acid sequence of SEQ ID NO: 3.
  12. 12. The expression vector of claim 1, wherein the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3.
  13. 13. The expression vector of claim 1, wherein the PMCV polyprotein comprises a first polyprotein at least 90% identical to SEQ ID NO: 1 and a second polyprotein, wherein the second polyprotein is at least 90% identical to SEQ ID NO: 2 or is at least 90% identical to SEQ ID NO: 3.
  14. 14. The expression vector of claim 1, wherein the PMCV polyprotein comprises i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3.
  15. 15. The expression vector of claim 1, wherein the PMCV polyprotein consists of i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3.
  16. 16. The expression vector of claim 1, wherein the PMCV polyprotein consists essentially of i) SEQ ID NO: 1 and ii) either SEQ ID NO: 2 or SEQ ID NO: 3.
  17. 17. A polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and any combination thereof.
  18. 18. The polypeptide of claim 17, wherein the amino acid sequence comprises SEQ ID NO: 1.
  19. 19. The polypeptide of claim 17, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 1.
  20. 20. The polypeptide of claim 17, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 1.

Description

COMPOSITIONS FOR PREVENTION OF CARDIOMYOPATHY SYNDROME CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63/523,471, filed on June 27, 2023, the entire disclosure of which is incorporated herein by reference. TECHNICAL FIELD The disclosure relates to constructs and compositions directed to immunization of animals to aid in the prevention of cardiomyopathy syndrome (CMS) caused by Piscine Myocarditis Virus (PMCV). In one aspect, the present disclosure provides compositions and methods directed to protection of salmon. BACKGROUND AND SUMMARY OF THE INVENTION Cardiomyopathy syndrome (CMS) is an emerging disease of economic importance in salmon. In particular, the Piscine Myocarditis Virus (PMCV) has been identified as a causative link for CMS. PMCV is of large economic concern in the salmonid industry because it causes morbidity and mortality in adult fish that are nearly full grown and almost ready for harvest. PMCV encodes at least three major proteins: a capsid (ORF1), a RNA- dependent RNA polymerase (ORF2), and a structural protein of unknown function (ORF3) that may be involved in virus assembly or egress. However, attempts to propagate PMCV using in vitro cell culture systems have failed. As a result, traditional live, attenuated, and inactivated vaccine approaches have not yet materialized. Thus, there exists a need for a PMCV vaccine that utilizes DNA, RNA, or recombinant approaches for design. Accordingly, the present disclosure provides constructs and compositions capable of immunizing animals against PMCV. The exemplary embodiments of the present disclosure include two different approaches of DNA-based vaccination. Using nanoplasmid technology, a series of plasmids were designed as DNA vaccine vectors. The series includes exemplary plasmids such as a plasmid including ORF1, a plasmid including ORF3, and a dual expression plasmid including both ORF1 and ORF3. The constructs and compositions of the present disclosure can provide beneficial effects against CMS for salmon. BRIEF DESCRIPTIONS OF THE DRAWINGS The detailed description particularly refers to the accompanying figures in which: FIGURE 1 shows atrium scores (0-4) for the different groups at 12 weeks postchallenge. FIGURE 2 shows ventricle scores (0-4) for the different groups at 12 weeks post-challenge. FIGURE 3 shows the mean ±95% CI for the sum of scores (severity) for atrium + ventricle is shown for all groups at 12 weeks post-challenge. A distinct separation was observed between groups at 12 weeks post-challenge. FIGURE 4 shows Ct values for heart in different groups at 12 weeks postchallenge. Results are shown as mean and 95% CI (spikes). DETAILED DESCRIPTION Various embodiments are described herein as follows. In an illustrative aspect, an expression vector is provided. The expression vector encodes i) a salmonid alphavirus (SAV) polyprotein and ii) a piscine myocarditis virus (PMCV) polyprotein. Salmonid alphavirus (SAV) is known in the art as an enveloped, single-stranded, positive-sense RNA virus with a genome of about 12 kb. For instance, Clynav™ is a DNA vaccine that can be used to protect Atlantic salmon against pancreatic disease caused by SAV. Certain aspects regarding Clynav™ are found, for example, in U.S. Patent No. 9,303,071, which is herein incorporated by reference in its entirety. In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 2 (SAV2) polyprotein. In an embodiment, the salmonid alphavirus polyprotein is a salmonid alphavirus subtype 3 (SAV3) polyprotein. In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 3. As described herein, SEQ ID NO: 3 corresponds to the ORF_3 (wild type) sequence. In an embodiment, the PMCV polyprotein is at least 91% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 92% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 93% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 94% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 95% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 96% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 97% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 98% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 99% identical with an amino acid sequence of SEQ ID NO: 3. In an embodiment, the PMCV polyprotein is at least 90% identical with an amino acid sequence of SEQ ID NO: 2. As described herein, SEQ ID NO: 2 corresponds to the ORF_3 (mutant)