Conjugated virus-like particles and uses thereof as anti-tumor immune redirectors
Disclosed is a new class of conjugated virus-like particles (VLPs). These conjugated VLPs bind a wide variety of tumors and comprise epitopes recognized by a prior T cell immune response already existing in a host. These epitopes are derived from pathogens or previous vaccinations (such as early childhood vaccines). This provokes the body's pre-existing cytotoxic immunity obtained through previous infection or previous childhood vaccination to be redirected to the tumor cells for the elimination of cancer, and form long-term anti-tumor immunity. The described conjugated VLPs are useful for tailoring a broad range of tumors towards a response from existing immunity circumventing the need to identify tumor antigens or generate tumor-specific immune responses. Importantly, the compositions and methods described herein broadens opportunities for treatment for all cancer types in subjects who previously had un-targetable cancers due to various technological and biological limitations of currently available immuno-therapeutic drugs.
1. A virus-like particle (VLP), consisting of:
at least one virus capsid protein, and
a fusion protein comprising in order from amino terminus to carboxy terminus as a single peptide:
a) one protease cleavage peptide sequence, attached directly to
b) at least one recall protein comprising at least one epitope,
wherein the at least one virus capsid protein is conjugated to one or more of the fusion protein through a disulfide bond, an ester bond, an amide bond, or a chemical linkage created by exposure of the at least one recall protein and at least one capsid protein simultaneously to a cross-linking agent under conditions amenable to formation of a chemical cross-link,
wherein the fusion protein is 45 amino acids or less in length,
wherein the one protease cleavage peptide sequence is recognized by a tumor microenvironment protease selected from one or more of: cathepsins, kallikreins, serine proteases, caspases, matrix metalloproteinases, disintegrin, and metalloproteinases (ADAMs), and
wherein the at least one epitope is at least one human T cell epitope that is from 8 to 17 amino acids in length.
2. The VLP of claim 1 , wherein the at least one epitope:
is a pathogen epitope,
comprises a peptide sequence that complexes with an MHC molecule, and/or
has a sequence selected from one or more of SEQ ID NOS:1 to 83.
3. The VLP of claim 1 , wherein the capsid protein is a papilloma virus L1 and/or L2 capsid protein.
4. The VLP of claim 3 , wherein the L1 protein is from Bovine papilloma virus (BPV), Human papilloma virus (HPV), Rabbit papilloma virus (RPV), or Mouse papilloma virus (MPV).
5. The VLP of claim 3 , wherein the L1 protein is from a bacteriophage or from a plant.
6. The VLP of claim 1 , wherein the fusion protein comprises at least two recall proteins.
7. The VLP of claim 1 , wherein the at least one capsid protein exhibits tropism for a specific type of tissue.
8. The VLP of claim 7 , wherein the tropism is a tropism for cells or tissues expressing heparin sulfate proteoglycan (HSPG).
9. The VLP of claim 1 , wherein the capsid protein is from human papilloma virus, hepatitis B virus, bacteriophage MS2, bacteriophage Qβ, bacteriophage P22, cowpea chlorotic mottle virus, cowpea mosaic virus, influenza virus, parvovirus, Norwalk virus, hamster polyoma virus, Macrobrachium rosenbergii nodavirus , hepatitis C virus, or a retrovirus.
10. The VLP of claim 1 , wherein the epitope sequence is from a childhood vaccine.
11. The VLP of claim 2 , wherein the at least one epitope is a viral epitope and is from vaccinia virus, varicella zoster virus, herpes zoster virus, rubella, hepatitis virus, influenza virus, measles virus, mumps virus, poliovirus, variola virus, rabies virus, dengue virus, Ebola virus, West Nile virus, yellow fever virus, zika virus, cytomegalovirus, or Epstein-Barr virus.
12. The VLP of claim 2 , wherein the at least one epitope is a bacterial epitope and is from Bordetella pertussis, Clostridium tetani, Chlamydia trachomatis , diphtheria, Hemophilus influenza, Meningococcus, Pneumococcus, Vibrio cholera, Mycobacterium tuberculosis, Bacillus Calmette-Guérin (BCG), typhoid, Escherichia coli, Salmonella, Legionella pneumophila, Rickettsia, Treponema pallidum pallidum, Streptococcus, Bacillus anthracis, Clostridium botulinum , or Yersinia.
13. The VLP of claim 2 , wherein the at least one epitope is a parasitic epitope and is from Entamoeba histolytica, Toxoplasma gondii, Trichinella, Trichomonas, Trypanosoma , or Plasmodium.
14. The VLP of claim 1 , wherein the fusion protein is conjugated to one capsid protein via a cysteine, lysine, or arginine residue.
15. The VLP of claim 14 , wherein about 20 to about 100 percent of the cysteine, lysine, and/or arginine residues of the at least one capsid protein are conjugated to the fusion protein.
16. A virus-like particle (VLP), comprising:
at least one virus capsid protein, and
a fusion protein consisting of, in order from amino terminus to carboxy terminus as a single peptide:
a) one protease cleavage peptide sequence, attached to
b) one recall protein comprising at least one epitope,
wherein the fusion protein is 45 amino acids or less in length,
wherein the protease cleavage peptide sequence is recognized by a tumor microenvironment protease selected from one or more of: cathepsins, kallikreins, serine proteases, caspases, matrix metalloproteinases, disintegrin and metalloproteinases (ADAMs),
wherein the at least one epitope is from 8 to 17 amino acids in length, and
wherein the at least one capsid protein exhibits tropism for a specific type of tissue.
17. The VLP of claim 16 , wherein the at least one epitope is a viral epitope from vaccinia virus, varicella zoster virus, herpes zoster virus, rubella, hepatitis virus, influenza virus, measles virus, mumps virus, poliovirus, variola virus, rabies virus, dengue virus, Ebola virus, West Nile virus, yellow fever virus, zika virus, cytomegalovirus, or Epstein-Barr virus.
18. The VLP of claim 1 , wherein the at least one epitope is heterologous to the at least one virus capsid protein.
19. The VLP of claim 16 , wherein the at least one epitope is heterologous to the at least one virus capsid protein.