Virus-inspired compositions and methods of redirecting preexisting immune responses using the same for treatment of cancer
Disclosed are virus-inspired compositions and preparation methods thereof, where the compositions comprise mutant papillomavirus L1 proteins that spontaneously form capsid backbones and that are conjugated to a peptide comprising an epitope to form immune redirector capsids (IRCs). The epitopes on the peptides are designed to be recognized by a subject's immune system based on the subject's preexisting immune memory developed from the subject's past exposure to the epitope through infection or vaccination. The mutant papillomavirus L1 proteins possess three mutations including an amino-terminal truncation, a carboxy-terminal truncation, and a truncation at helix four. These mutations in the L1 protein yield capsomeres that are form non-canonical T=1 geometry capsid backbones. Disclosed are uses and methods of using the compositions in treating and/or preventing cancers in subjects in need thereof.
1. A composition, comprising:
a plurality of virus proteins, wherein each of said plurality of virus proteins comprises a mutated amino acid sequence of a Papillomaviridae L1 protein;
one or more peptides each comprising one or more epitopes from one or more pathogens other than a Papillomaviridae antigenic peptide;
wherein the mutated amino acid sequence of the Papillomaviridae L1 protein comprises at least the following mutations with respect to the wild type L1 protein sequence: (a) a deletion of at least five amino acid residues from an amino-terminus, and (b) a deletion of at least ten amino acid residues from the helix four region,
wherein the one or more peptides are attached to the plurality of virus proteins, and
wherein said plurality of virus proteins spontaneously assemble to form an icosahedron or dodecahedron capsid having a triangulation number T equal to 1 that binds to proteoglycan expressed on tumor cells.
2. The composition of claim 1 , wherein the amino acid sequence of each of the plurality of mutant Papillomaviridae L1 proteins further comprises: (c) a deletion of at least thirty amino acid residues from a carboxy-terminus.
3. The composition of claim 1 , wherein the one or more peptides are conjugated to the plurality of mutant Papillomaviridae L1 proteins.
4. The composition of claim 3 , wherein the one or more peptides are conjugated to the mutant Papillomaviridae L1 proteins via a cysteine, lysine, or arginine residue of the mutant Papillomaviridae L1 protein.
5. The composition of claim 3 , wherein the one or more peptides are conjugated to the mutant Papillomaviridae L1 proteins via disulphide, maleimide, or amide bond between the mutant Papillomaviridae L1 protein and a residue of the peptide.
6. The composition of claim 1 , wherein at least 25% of the plurality of mutant Papillomaviridae L1 proteins are conjugated to at least one of the one or more peptides.
7. The composition of claim 1 , wherein at least 25 to 85% (w/w) of the plurality of mutant Papillomaviridae L1 proteins are conjugated to at least one of the one or more peptides.
8. The composition of claim 1 , wherein each of the one or more peptides further comprise one or more protease cleavage sequences.
9. The composition of claim 8 , wherein the one or more protease cleavage sequences comprise a furin cleavage sequence, a matrix metalloprotease cleavage sequence, or a disintegrin and metalloprotease (ADAM) cleavage sequence.
10. The composition of claim 1 , wherein the one or more epitopes are viral, bacterial, parasitic, or fungal epitopes.
11. The composition of claim 10 , wherein:
the viral epitopes are one or more of coronavirus, vaccinia, Varicella zoster, Herpes zoster, rubella, hepatitis, influenza, measles, mumps, poliovirus, variola, rabies, dengue, Ebola, West Nile, yellow fever, or zika epitopes;
the bacterial epitopes are one or more of Bordetella pertussis, Clostridium tetani, Chlamydia trachomatis, Corynebacterium diphtheriae, Hemophilus influenza, Neisseria meningitidis, Streptococcus, Vibrio cholera, Mycobacterium tuberculosis, Bacillus Calmette-Guérin, Salmonella, Escherichia coli, Legionella pneumophila, Rickettsia, Treponema pallidum pallidum, Bacillus anthracis, Clostridium botulinum , or Yersinia epitopes; or
the parasitic epitopes are one or more of Entamoeba histolytica, Toxoplasma gondii, Trichinella, Trichomonas, Trypanosoma , or Plasmodium epitopes.
12. The composition of claim 1 , wherein at least one of the one or more epitopes is a childhood vaccine antigenic epitope.
13. The composition of claim 1 , wherein the one or more peptides comprises at least two epitopes from one or more pathogens other than a Papillomaviridae antigenic peptides.
14. The composition of claim 1 , wherein the proteoglycan expressed on tumor cells is heparin sulfate proteoglycan (HSPG), perlecan, hyalectan versican, glypican-3, small leucine-rich proteoglycans (SLRP), and/or biglycan.
15. The composition of claim 1 , wherein the plurality of mutant Papillomaviridae L1 proteins do not form a T=7 capsid backbone.
16. The composition of claim 1 , wherein said plurality of mutant Papillomaviridae L1 proteins are mouse mutant Papillomaviridae L1 proteins.
17. The composition of claim 1 , wherein an amino acid sequence of each of said plurality of mutant Papillomaviridae L1 proteins is SEQ ID NO:134, and is encoded by nucleic acid sequence SEQ ID NO:135 or 136.
18. A method of treating, reducing the occurrence of, inhibiting the progression and/or metastasis of, a cancer in a subject in need thereof, which comprises administering to the subject a pharmaceutically effective amount of a composition comprising:
a plurality of virus proteins, wherein each of said plurality of virus proteins comprises a mutated amino acid sequence of a Papillomaviridae L1 protein;
one or more peptides each comprising one or more epitopes from one or more pathogens other than a Papillomaviridae antigenic peptide;
wherein the mutated amino acid sequence of the Papillomaviridae L1 protein comprises at least the following mutations with respect to the wild type L1 protein sequence: (a) a deletion of at least five amino acid residues from an amino-terminus, and (b) a deletion of at least ten amino acid residues from the helix four region,
wherein the one or more peptides are attached to the plurality of virus proteins, and
wherein said plurality of virus proteins spontaneously assemble to form an icosahedron or dodecahedron capsid having a triangulation number T equal to 1 that binds to proteoglycan expressed on tumor cells.
19. The method of claim 18 , further comprising:
obtaining from the subject a tumor tissue sample; and
identifying in the tumor tissue a sequence of one or more MHC molecules expressed by one or more tumor cells in the tumor tissue sample.
20. The method of claim 18 , wherein the subject was previously infected or vaccinated against a pathogen, and wherein the one or more epitopes is an antigenic epitope of the pathogen.
21. The method of claim 18 , wherein the one or more epitopes are capable of complexing with one or more MHC molecules expressed by a tumor cell in a tumor tissue sample obtained from the subject.
22. A process for producing the composition of claim 1 , which comprises:
(a) transforming a prokaryotic cell with an expression vector encoding the L1 protein;
(b) culturing the transformed prokaryotic cell under conditions that promote expression of the L1 protein;
(c) lysing the transformed prokaryotic cells to release expressed L1 protein;
(d) separating cell debris from the expressed L1 protein and recovering the L1 protein as inclusion bodies;
(e) optionally washing the L1 protein inclusion bodies;
(f) solubilizing the L1 protein inclusion bodies;
(g) refolding the L1 protein; and
(h) forming the icosahedron or dodecahedron capsid having a triangulation number T equal to 1 by incubating the refolded L1 protein in refolding buffer.
23. The process of claim 22 , further comprising:
(i) conjugating in a conjugation buffer the one or more peptides to the assembled L1 protein by incubating the assembled L1 protein under reducing conditions in the presence of one or more peptides.
24. The process of claim 22 , wherein the refolding buffer comprises a denaturant, a reducing agent, 250 to 500 mM of a salt, a non-ionic surfactant, a metal chelating agent, and a buffer of pH 7.5 to 8.5.
25. The process of claim 24 , wherein the salt is present from 250 mM to 500 mM.
26. The process of claim 24 , wherein the molar ratio of the one or more peptides to the L1 protein is at least 1:5.
27. The process of claim 22 , further comprising removing denaturant from the refolding buffer but maintaining reducing agent when forming the icosahedron or dodecahedron capsid having a triangulation number T equal to 1.