Chaperonin 10 variants

Abstract

The invention relates generally to chaperonin 10 N-terminal variants. More specifically, the invention relates to chaperonin 10 N-terminal variants with enhanced immunomodulatory capacity and/or enhanced binding affinity for pathogen-associated molecular patterns (PAMPs) and/or damage-associated molecular patterns (PAMPs).

What is claimed is: 
     
         1 . An isolated chaperonin 10 (Cpn10) variant polypeptide derived from human Cpn10 sharing at least 90% sequence identity with a wild-type human Cpn10 polypeptide, wherein the polypeptide is selected from the group consisting of SEQ ID NO: 6 (Gly-Cpn10); SEQ ID NO: 11 (Cpn10-deltaNterm); SEQ ID NO: 63 (V-Cpn10); SEQ ID NO: 64 (L-Cpn10); SEQ ID NO: 65 (I-Cpn10); SEQ ID NO: 66 (H-Cpn10); SEQ ID NO: 67 (F-Cpn10); SEQ ID NO: 68 (Y-Cpn10); SEQ ID NO: 69 (W-Cpn10); SEQ ID NO: 70 (C-Cpn10); SEQ ID NO: 71 (T-Cpn10); SEQ ID NO: 72 (D-Cpn10); SEQ ID NO: 73 (N-Cpn10); SEQ ID NO: 74 (E-Cpn10); SEQ ID NO: 75 ((Q-Cpn10); SEQ ID NO: 76 (QS-Cpn10); SEQ ID NO: 77 (QSM-Cpn10); SEQ ID NO: 79 (Ala-Cpn10-deltaroof); and SEQ ID NO: 80 (Ala-Cpn10-K53M, K55M). 
     
     
         2 . An isolated nucleic acid molecule encoding a chaperonin 10 (Cpn10) variant polypeptide derived from human Cpn10 sharing at least 90% sequence identity with a wild-type human Cpn10 polypeptide and comprising an N-terminus extended by at least two to five additional amino acid residues compared to said wild-type polypeptide, wherein the polypeptide has increased binding affinity for a pathogen-associated molecular pattern (PAMP) compared to the binding affinity of Ala-Cpn10 (SEQ ID NO: 3) for said PAMP, wherein the isolated Cpn10 polypeptide possesses a conserved core antiparallel β-barrel flanked by a β-hairpin roof loop region and a mobile loop region. 
     
     
         3 . The isolated nucleic acid molecule of  claim 2  wherein the N-terminus of said human variant polypeptide commences with a methionine residue. 
     
     
         4 . The isolated nucleic acid molecule of  claim 3  wherein said methionine residue precedes an amino acid residue selected from the group consisting of arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, and valine. 
     
     
         5 . An isolated nucleic acid molecule encoding a chaperonin 10 (Cpn10) variant polypeptide sharing at least 90% sequence identity with a wild-type Cpn10 polypeptide and comprising an N-terminus extended by at least two additional amino acid residues compared to said wild-type polypeptide, wherein the polypeptide has increased binding affinity for a pathogen-associated molecular pattern (PAMP) compared to the binding affinity of Ala-Cpn10 (SEQ ID NO: 3) for said PAMP, wherein the N-terminus of said variant polypeptide commences with a methionine residue, and wherein said methionine residue precedes an amino acid residue selected from the group consisting of arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, and valine, wherein said polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 29. 
     
     
         6 . The isolated nucleic acid molecule of  claim 2  wherein said polypeptide comprises the amino acid sequence as set forth in any one of SEQ ID NOs: 52, 55, 60, 81, 84, 87, or 96. 
     
     
         7 . A method for the treatment of a disease or condition selected from the group consisting of rheumatoid arthritis, inflammatory bowel disease (Crohn's disease, ulcerative colitis), diabetes type I (insulin-dependent diabetes mellitus, juvenile onset diabetes), chronic fatigue syndrome, Alzheimer's disease, Graves' disease, osteoarthritis, collagen II arthritis, multiple sclerosis, systemic lupus erythematosus, autoimmune myocarditis, autoimmune ovarian disease, autoimmune thyroid disease, autoimmune neuritis, autoimmune hepatitis, autoimmune uveoretinitis, autoimmune uveitis, psoriasis, Sjögren's disease, sarcoidosis, dermatomyositis, leukocytoclastic vasculitis, myasthenia gravis, allergic encephalomyelitis, thyrotoxicosis, pernicious anemia, polymyalgia rheumatica, polymyositis, chronic obstructive pulmonary disease (COPD), infectious diseases, leaky gut syndrome, congestive heart disease, anaphylaxis, drug reactions, skin allergy, eczema, allergic rhinitis, urticaria, atopic dermatitis, allergic contact allergy, food allergy, allergic conjunctivitis, insect venom allergy, asthma, acute respiratory distress syndrome (ARDS), and atherosclerosis comprising the step of administering:
 (a) a therapeutically effective quantity of an isolated chaperonin 10 (Cpn10) variant polypeptide derived from human Cpn10 sharing at least 90% sequence identity with a wild-type human Cpn10 polypeptide and comprising an N-terminus extended by at least two to five additional amino acid residues compared to said wild-type polypeptide, wherein the polypeptide has increased binding affinity for a pathogen-associated molecular pattern (PAMP) compared to the binding affinity of Ala-Cpn10 (SEQ ID NO: 3) for said PAMP, wherein the isolated Cpn10 polypeptide possesses a conserved core antiparallel β-barrel flanked by a β-hairpin roof loop region and a mobile loop region; or 
 (b) a nucleic acid encoding chaperonin 10 (Cpn10) variant polypeptide derived from human Cpn10 sharing at least 90% sequence identity with a wild-type human Cpn10 polypeptide and comprising an N-terminus extended by at least two to five additional amino acid residues compared to said wild-type polypeptide, wherein the polypeptide has increased binding affinity for a pathogen-associated molecular pattern (PAMP) compared to the binding affinity of Ala-Cpn10 (SEQ ID NO: 3) for said PAMP, wherein the isolated Cpn10 polypeptide possesses a conserved core antiparallel β-barrel flanked by a β-hairpin roof loop region and a mobile loop region.