Novel galectin sequences and compositions and methods utilizing same for treating or diagnosing arthritis and other chronic inflammatory diseases

Abstract

Isolated polynucleotides encoding novel galectin-8 variants polypeptides are provided. Also provided are methods and kits using same for diagnosing prognosing and treating rheumatoid arthritis (RA) and other joint/chronic inflammatory diseases.

1 . A pharmaceutical composition, comprising, as an active ingredient, galectin-8 or a functional equivalent or derivative thereof and at least one type of a disease-modifying antirheumatic drug (DMARD) and a pharmaceutically acceptable carrier.  
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein said at least one type of a disease-modifying antirheumatic drug is selected from the group consisting of auranofin, azathioprine, cyclophosphamide, cyclosporine, gold sodium thiomalate, hydroxychloro-quine sulfate, leflunomide, methotrexate, minocycline, penicillamine, sulfasalazine, etanercept, infliximab, anakinra and adalimumab.  
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein said galectin-8 is as set forth in SEQ ID NO: 4.  
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein said galectin-8 variant is as set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         5 . A pharmaceutical composition, comprising, as an active ingredient, galectin-8 or a functional equivalent or derivative thereof and at least one type of a nonsteroidal anti-inflammatory drug (NSAID) and a pharmaceutically acceptable carrier.  
     
     
         6 . The pharmaceutical composition of  claim 5 , wherein said galectin-8 is as set forth in SEQ ID NO: 4.  
     
     
         7 . The pharmaceutical composition of  claim 5 , wherein said galectin-8 variant is as set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         8 . The pharmaceutical composition of  claim 5 , wherein said at least one type of a nonsteroidal anti-inflammatory drug (NSAID) is selected from the group consisting of diclofenac potassium, diclofenac sodium, diclofenac sodium with misoprostol, diflunisal, etodolac, fenoprofen calcium, flurbiprofen, ibuprofen, indomethacin, ketoprofen, meclofenamate sodium, mefenamic acid, meloxicam, nabumetone, naproxen, naproxen sodium, oxaprozin, piroxicam, sulindac, tolmetin sodium, celecoxib, rofecoxib, valdecoxib, aspirin, choline and magnesium salicylates, choline salicylate, magnesium salicylate, salsalate, sodium salicylate, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisolone sodium phosphate and prednisone.  
     
     
         9 . An isolated polypeptide comprising a galectin-8 sequence being devoid of the amino acid sequence region encompassed by amino acid coordinates 115-150 of SEQ ID NO: 4.  
     
     
         10 . The isolated polypeptide of  claim 9 , wherein said galectin-8 sequence is set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         11 . A pharmaceutical composition, comprising, as an active ingredient, a galectin-8 sequence devoid of the amino acid sequence region encompassed by amino acid coordinates 115-150 of SEQ ID NO: 4 and a pharmaceutically acceptable carrier.  
     
     
         12 . The pharmaceutical composition of  claim 11 , wherein said galectin-8 sequence is set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         13 . An article-of-manufacture comprising packaging material and a pharmaceutical composition identified for reducing inflammatory response, being contained within said packaging material, said pharmaceutical composition including, as an active ingredient, a galectin-8 or a variant or a derivative thereof and a pharmaceutically acceptable carrier.  
     
     
         14 . The article-of-manufacture of  claim 3 , wherein said galectin-8 is set forth in SEQ ID NO: 4.  
     
     
         15 . The article-of-manufacture of  claim 3 , wherein said galectin-8 variant is set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         16 . A method of reducing inflammatory response in an individual, comprising providing to the individual a galectin-8 polypeptide or a variant or derivative thereof, thereby reducing the inflammatory response in the individual.  
     
     
         17 . The method of  claim 16 , wherein said galectin-8 is SEQ ID NO:4.  
     
     
         18 . The method of  claim 16 , wherein said variant is SEQ ID NO: 6, 8 or 16.  
     
     
         19 . The method of  claim 16 , wherein said providing is effected by expressing said polypeptide within cells of the individual.  
     
     
         20 . The method of  claim 19 , wherein said cells of the individual are synovial fluid cells.  
     
     
         21 . The method of  claim 16 , wherein said providing is effected by administration of a pharmaceutical composition comprising a galectin-8 or a variant or a derivative thereof.  
     
     
         22 . The method of  claim 21 , wherein said administration is oral, injection or topical administration.  
     
     
         23 . The method of  claim 22 , wherein said administration by injection is direct injection into a joint of the individual.  
     
     
         24 . The method of  claim 16 , wherein said providing is effected by: 
 (i) isolating synovial fluid cells from the individual;    (ii) transforming said synovial fluid cells with an expression construct capable of expressing said galectin-8 polypeptide or said variant or derivative thereof in said synovial fluid cells; and    (iii) administering said synovial fluid cells resultant from step (b) into the individual.    
     
     
         25 . A population of cells comprising synovial fluid cells transformed to express galectin-8 or a functional equivalent or a variant or derivative thereof.  
     
     
         26 . The pharmaceutical composition of  claim 25 , wherein said galectin-8 is as set forth in SEQ ID NO: 4.  
     
     
         27 . The pharmaceutical composition of  claim 25 , wherein said galectin-8 variant is as set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         28 . An isolated polynucleotide encoding a polypeptide comprising a galectin-8 sequence being devoid of the amino acid sequence region encompassed by amino acid coordinates 115-150 of SEQ ID NO: 4.  
     
     
         29 . The isolated polynucleotide of  claim 28  wherein the polynucleotide is set forth in SEQ ID NO: 5, 7 or 15.  
     
     
         30 . A nucleic acid construct comprising the polynucleotide of  claim 28 .  
     
     
         31 . The nucleic acid construct of  claim 30 , further comprising a promoter for regulating expression of the polynucleotide.  
     
     
         32 . A cell comprising the nucleic acid construct of  claim 31 .  
     
     
         33 . A method of detecting an inflammatory response in an individual comprising identifying in biological sample obtained from the individual a transcription and/or translation product of a galectin-8 variant, wherein a presence of said transcription and/or translation product is indicative of an inflammatory response.  
     
     
         34 . The method of  claim 33 , wherein said biological sample is the synovial fluid cells.  
     
     
         35 . The method of  claim 33 , wherein said galectin-8 variant is set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         36 . A method of diagnosing an individual having an inflammatory disease comprising: 
 (a) obtaining a biological sample from the individual; and    (b) detecting in said biological sample a transcription and/or translation product of a galectin-8 variant, wherein a presence of said transcription and/or translation product indicates presence of an inflammatory disease.    
     
     
         37 . The method of  claim 36 , wherein said biological sample includes synovial fluid cells.  
     
     
         38 . The method of  claim 36 , wherein said galectin-8 variant is set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         39 . A pair of oligonucletides capable of directing PCR amplification of a polynucleotide sequence encoding a galectin-8 variant.  
     
     
         40 . The pair of oligonucletides of  claim 39 , wherein said pair of oligonucletides are set forth in SEQ ID NOS: 1 and 2, SEQ ID NOs: 11 and 12 or SEQ ID NOs: 12 and 14.  
     
     
         41 . The pair of oligonucletides of  claim 39 , wherein said galectin-8 variant is encoded by SEQ ID NO: 5, 7 or 15.  
     
     
         42 . A kit for diagnosing RA patients comprising at least one pair of nucleotides of  claim 39  and reagents for detecting a PCR product.  
     
     
         43 . A polynucleotide probe comprising a nucleic acid sequence specifically hybridizable with a polynucleotide sequence encoding a galectin-8 variant and not a galectin-8 as set forth in SEQ ID NO: 3.  
     
     
         44 . The polynucleotide probe of  claim 43 , wherein said polynucleotide sequence encoding galectin-8 variant is SEQ ID NO: 5, 7 or 15.  
     
     
         45 . A kit for diagnosing RA patients comprising at least one polynucleotide probe of  claim 43  and reagents for detecting a hybridization product.  
     
     
         46 . The kit of  claim 45 , wherein said at least one polynucleotide probe is labeled with a detectable moiety.  
     
     
         47 . The kit of  claim 46 , wherein said detectable moiety is selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a light-emitting moiety and a radioactive moiety.  
     
     
         48 . An antibody comprising an antigen recognition sequence capable of specifically binding a galectin-8 variant and not a galectin-8 as set forth in SEQ ID NO: 4.  
     
     
         49 . The antibody of  claim 48 , wherein said galectin-8 variant is set forth in SEQ ID NO: 6, 8 or 16.  
     
     
         50 . A kit for diagnosing RA patients, comprising the antibody of  claim 48  and reagents for detecting antibody binding.  
     
     
         51 . The kit of  claim 50 , wherein said polynucleotide probe is labeled with a detectable moiety.  
     
     
         52 . The kit of  claim 51 , wherein said detectable moiety is selected from the group consisting of a chromogenic moiety, a fluorogenic moiety, a light-emitting moiety and a radioactive moiety.