Engineered HCV E2 immunogens and related vaccine compositions
The present invention provides novel engineered HCV E2 polypeptide immunogens and related vaccine compositions that display the engineered E2 polypeptides. The invention also provides methods of using such immunogens and vaccine compositions in various therapeutic applications, e.g., for preventing or treating HCV infections.
1. A modified HCV E2 ectodomain polypeptide, comprising an engineered E2 ectodomain sequence that contains a truncation in the VR2 disordered region, wherein the truncated VR2 disordered region contains less than 20 amino acid residues.
2. The modified HCV E2 ectodomain polypeptide of claim 1 , wherein the VR2 disordered region corresponds to amino acid residues 452-494 of the full length polyprotein of HCV H77 isolate.
3. The modified HCV E2 ectodomain polypeptide of claim 2 , wherein truncation of the VR2 disordered region comprises replacing residues 41-61 of SEQ ID NO:4, CPERLASCGSSGCWHYPPRPC (SEQ ID NO:6), with CPERASGHYPRPC (SEQ ID NO:10).
4. The modified HCV E2 ectodomain polypeptide of claim 2 , wherein truncation of the VR2 disordered region comprises replacing residues 41-61 of SEQ ID NO:4, CPERLASCGSSGCWHYPPRPC (SEQ ID NO:6), with a sequence set forth in CXXXXXXHYPRPC (SEQ ID NO:8) or CXXXXXHYPRPC (SEQ ID NO:9), wherein X is any amino acid residue.
5. The modified HCV E2 ectodomain polypeptide of claim 4 , wherein XXXXXX in SEQ ID NO:8 is QNWDEP (SEQ ID NO:11), KVNIDP (SEQ ID NO:12), EKVEEL (SEQ ID NO:13), PDENMK (SEQ ID NO:14), or KREEKM (SEQ ID NO:15).
6. The modified HCV E2 ectodomain polypeptide of claim 4 , wherein XXXXX in SEQ ID NO:9 is PKTEV (SEQ ID NO:16), KRVDI (SEQ ID NO:17), PSDMV (SEQ ID NO:18), PNEEE (SEQ ID NO:19), or KKEIR (SEQ ID NO:20).
7. The modified HCV E2 ectodomain polypeptide of claim 1 , further comprising a deletion in the β-sandwich loop connecting βsheets 6 and 7 of the β-sandwich domain.
8. The modified HCV E2 ectodomain polypeptide of claim 7 , wherein deletion of the β-sandwich loop sequence (SEQ ID NO:21) comprises deletion of one or more residues that form the tip of the β-sandwich loop.
9. The modified HCV E2 ectodomain polypeptide of claim 8 , wherein residues that form the tip of the β-sandwich loop are residues 543-546 from HCV genotype 1a H77 isolate polyprotein sequence.
10. The modified HCV E2 ectodomain polypeptide of claim 8 , wherein residues 543-546 from HCV genotype 1a H77 isolate polyprotein sequence, RPPL (SEQ ID NO:22), are deleted.
11. The modified HCV E2 ectodomain polypeptide of claim 7 , comprising an amino acid sequence as set forth in any one of SEQ ID NOs:26-38, a conservatively modified variant or a substantially identical sequence thereof.
12. A polynucleotide encoding the modified HCV E2 ectodomain polypeptide of claim 1 .
13. A pharmaceutical composition, comprising the modified HCV E2 ectodomain polypeptide of claim 1 , and a pharmaceutically acceptable carrier.
14. An immunogenic composition, comprising a modified HCV E2 ectodomain polypeptide of claim 1 that is displayed on the surface of a self-assembling nanoparticle.
15. The immunogenic composition of claim 14 , wherein the C-terminus of the modified HCV E2 ectodomain polypeptide is fused to the N-terminus of a subunit of the self-assembling nanoparticle via a linker sequence.
16. The immunogenic composition of claim 15 , wherein the linker sequence comprises (GGGGS)2 (SEQ ID NO:42).
17. The immunogenic composition of claim 14 , wherein the modified HCV E2 ectodomain polypeptide comprises an amino acid sequence as set forth in any one of SEQ ID NOs:26 38.
18. The immunogenic composition of claim 14 , wherein the subunit of the self-assembling nanoparticle comprises the polypeptide as shown in SEQ ID NO:39 (E2p), SEQ ID NO: 40 (I3-01), or SEQ ID NO: 41 (ferritin).
19. A polynucleotide, encoding a fusion protein comprising a modified HCV E2 ectodomain polypeptide of claim 1 and a self-assembling nanoparticle subunit, wherein the modified HCV E2 ectodomain polypeptide is fused at its C-terminus to the N-terminus of the self-assembling nanoparticle subunit.
20. A pharmaceutical composition, comprising the immunogenic composition of claim 14 , and a pharmaceutically acceptable carrier.
21. A method of treating HCV infection in a subject, comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of the immunogenic composition of claim 14 , thereby treating HCV infection in the subject.