Biomarkers of osteoarthritis

Abstract

Biomarkers, biomarker panels and methods for diagnosing osteoarthritis (OA) are disclosed, using measurement of the expression level of certain polypeptides in a test sample from a subject, including MCP1, IL8, KC, MMP2, MMP3, IL6, MMP1, RANTES, MMP9, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP. Related methods for monitoring OA treatment efficacy, diagnostic reagents, and kits are also described.

1 . A method for diagnosing, staging, or monitoring osteoarthritis in a subject comprising: measuring in a biological sample from the subject the level of expression of at least two polypeptides selected from the group consisting of: MCP1, IL8, KC, MMP2, MMP3, MMP9, IL6, MMP1, RANTES, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP, and fragments of any thereof, and any combination thereof, wherein the expression levels of the at least two polypeptides or fragments thereof in the biological sample provide a sample protein expression profile indicative of the presence or absence, degree, severity, type or stage of osteoarthritis in the subject. 
     
     
         2 . The method of  claim 1 , wherein the biological sample comprises any one of synovial fluid, whole blood, blood plasma, serum, urine, and saliva. 
     
     
         3 . The method of  claim 1 , wherein the biological sample comprises synovial fluid. 
     
     
         4 . The method of  claim 1 , wherein the level of expression of at least four polypeptides is measured. 
     
     
         5 . The method of  claim 1 , wherein the subject is a human. 
     
     
         6 . The method of  claim 1 , wherein the subject is a canine 
     
     
         7 . The method of  claim 1 , comprising measuring in a biological sample from the subject the level of expression of MCP1, IL8, KC, MMP2 and MMP3, or fragments thereof. 
     
     
         8 . The method of  claim 7 , wherein the subject is a canine 
     
     
         9 . The method of  claim 1 , comprising measuring in a biological sample from the subject the level of expression of MCP1, IL6, IL8, KC and MMP1, or fragments thereof 
     
     
         10 . The method of  claim 9 , wherein the subject is a human. 
     
     
         11 . The method of  claim 10 , further comprising measuring in the biological sample the level of expression of RANTES, or fragments thereof. 
     
     
         12 . The method of  claim 1 , further comprising: comparing the sample protein expression profile to a control protein expression profile, wherein a difference between the sample protein expression profile and the control protein expression profile is indicative of the presence or absence, degree, severity, type or stage of osteoarthritis in the subject. 
     
     
         13 . The method of  claim 1 , wherein the subject is at risk of having or is suspected of having osteoarthritis. 
     
     
         14 . The method according to  claim 1 , wherein the level of expression of the at least two polypeptides in the biological sample is measured using a method selected from the group consisting of: LUMINEX, ELISA, immunoassay, mass spectrometry, high performance liquid chromatography, two-dimensional electrophoresis, qPCR, RT-PCR, nucleic acid microarray, in situ hybridization, SAGE, Western blotting, protein microarray, and antibody microarray. 
     
     
         15 . The method of  claim 1 , wherein the level of expression of the at least two polypeptides in the biological sample is measured by detecting alterations in DNA due to a process selected from the group consisting of DNA amplification, DNA methylation/demethylation, and single nucleotide polymorphisms. 
     
     
         16 . An OA biomarker expression profile comprising polypeptide expression level information for two or more polypeptides selected from the group consisting of: MCP1, IL8, KC, MMP2, MMP3, IL6, MMP1, RANTES, MMP9, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP and fragments of any thereof, and any combination thereof, obtained from a biological sample from a subject suspected of having osteoarthritis. 
     
     
         17 . The OA expression profile of  claim 15 , further comprising polypeptide expression level information for at least one biological sample obtained from at least one healthy subject. 
     
     
         18 . The OA expression profile of  claim 16 , wherein the biological samples from the subject suspected of having osteoarthritis and the healthy subject or subjects both comprise a sample of synovial fluid, a sample of whole blood, a sample of blood plasma, a sample of serum, a sample of urine, and a sample of saliva. 
     
     
         19 . The OA expression profile of  claim 16 , wherein the biological samples from the subject suspected of having osteoarthritis and the healthy subject or subjects both comprise a sample of synovial fluid. 
     
     
         20 . A diagnostic reagent for osteoarthritis comprising two or more antibodies against any two or more OA biomarkers or fragments thereof selected from the group consisting of: MCP1, IL8, KC, MMP2, MMP3, IL6, MMP1, RANTES, MMP9, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP and fragments of any thereof. 
     
     
         21 . A kit comprising the diagnostic reagent of  claim 19 . 
     
     
         22 . A kit for diagnosing osteoarthritis in a subject, the kit comprising: at least two OA biomarker detection reagents that each specifically bind to an OA polypeptide selected from the group consisting of MCP1, IL8, KC, MMP2, MMP3, IL6, MMP1, RANTES, MMP9, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP and fragments of any thereof, or at least two OA biomarker detection reagents that each specifically bind to at least part of a polynucleotide sequence coding for at least two of the OA polypeptides, wherein the specific binding of the reagent is indicative of the expression level of at least one OA polypeptide in a biological sample from a subject. 
     
     
         23 . The kit of  claim 21 , wherein said at least one reagent that specifically detects expression of at least one biomarker comprises a nucleic acid probe complementary to at least part of a polynucleotide sequence coding for one of the polypeptides. 
     
     
         24 . The kit of  claim 22 , wherein the nucleic acid probe is a cDNA or is an oligonucleotide. 
     
     
         25 . The kit of  claim 21 , wherein the at least one OA biomarker detection reagent is immobilized on a substrate surface. 
     
     
         26 . The kit of  claim 24 , comprising at least two biomarker detection reagents arranged on the substrate surface. 
     
     
         27 . The kit of  claim 25 , wherein the at least two biomarker reagents arranges on the substrate surface comprise a microarray. 
     
     
         28 . A method for identifying a candidate substance as a therapeutic agent for treating osteoarthritis, comprising: a) administering the candidate substance to a subject diagnosed with spontaneous osteoarthritis; b) measuring the expression level of two or more OA polypeptides selected from the group consisting of MCP1, IL8, KC, MMP2, MMP3, IL6, MMP1, RANTES, MMP9, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP in a biological sample from the subject; and c) selecting the candidate substance as a candidate therapeutic agent for treating osteoarthritis if the expression level of each of the two or more OA polypeptides in the biological sample is lower than or equal to the expression level for the selected two or more OA polypeptides in a biological sample from a control subject not administered the test substance. 
     
     
         29 . The method of  claim 27 , wherein the biological sample comprises any one of synovial fluid, whole blood, blood plasma, serum, urine, and saliva. 
     
     
         30 . The method of  claim 27 , wherein the biological sample comprises synovial fluid. 
     
     
         31 . The method of  claim 27 , wherein the level of expression of at least four polypeptides is measured. 
     
     
         32 . The method of  claim 27 , wherein the subject is a human. 
     
     
         33 . The method of  claim 27 , wherein the subject is a canine 
     
     
         34 . The method of  claim 27 , comprising measuring in a biological sample from the subject the level of expression of MCP1, IL8, KC, MMP2 and MMP3, or fragments thereof. 
     
     
         35 . The method of  claim 34 , wherein the subject is a canine. 
     
     
         36 . The method of  claim 27 , comprising measuring in a biological sample from the subject the level of expression of MCP1, IL6, IL8, KC and MMP1, or fragments thereof. 
     
     
         37 . The method of  claim 36 , wherein the subject is a human. 
     
     
         38 . The method of  claim 27 , wherein the expression level of each of the at least two polypeptides in the biological sample from the subject is measured using a method selected from the group consisting of: LUMINEX, ELISA, immunoassay, mass spectrometry, high performance liquid chromatography, two-dimensional electrophoresis, qPCR, RT-PCR, nucleic acid microarray, in situ hybridization, SAGE, Western blotting, protein microarray, and antibody microarray. 
     
     
         39 . A method for monitoring the effect of a treatment of osteoarthritis in a subject comprising: a) obtaining a first OA biomarker expression profile comprising measuring the expression level of two or more OA polypeptides selected from the group consisting of MCP1, IL8, KC, MMP2, MMP3, IL6, MMP1, RANTES, MMP9, IL1B, Apolipoprotein A1, Apolipoprotein E, DCN, CILP and COMP in a first biological sample obtained from the subject before the osteoarthritis treatment is administered to the subject; b) obtaining a second OA biomarker expression profile comprising measuring the expression level of the two or more OA polypeptides selected in (a), in a second biological sample obtained from the subject after or while the osteoarthritis treatment is administered to the subject; and c) comparing the first OA biomarker expression profile with the second OA biomarker expression profile, wherein if the expression level of each of the two or more selected OA polypeptides in the first OA biomarker expression profile is lower than or equal to the expression level for the selected two or more OA polypeptides in the second biological sample from the subject is indicative of a therapeutic effect of the osteoarthritis treatment in the subject. 
     
     
         40 . The method of  claim 38 , wherein the biological sample comprises any one of synovial fluid, whole blood, blood plasma, serum, urine, and saliva. 
     
     
         41 . The method of  claim 38 , wherein the biological sample comprises synovial fluid. 
     
     
         42 . The method of  claim 38 , wherein the level of expression of at least four polypeptides is measured. 
     
     
         43 . The method of  claim 38 , wherein the subject is a human. 
     
     
         44 . The method of  claim 38 , wherein the subject is a canine. 
     
     
         45 . The method of  claim 38 , comprising measuring in a biological sample from the subject the level of expression of MCP1, IL8, KC, MMP2 and MMP3, or fragments thereof. 
     
     
         46 . The method of  claim 45 , wherein the subject is a canine. 
     
     
         47 . The method of  claim 38 , comprising measuring in a biological sample from the subject the level of expression of MCP1, IL6, IL8, KC and MMP1, or fragments thereof. 
     
     
         48 . The method of  claim 47 , wherein the subject is a human. 
     
     
         49 . The method of  claim 38 , wherein the expression level of the at least two polypeptides in the biological sample from the subject is measured using a method selected from the group consisting of: LUMINEX, ELISA, immunoassay, mass spectrometry, high performance liquid chromatography, two-dimensional electrophoresis, qPCR, RT-PCR, nucleic acid microarray, in situ hybridization, SAGE, Western blotting, protein microarray, and antibody microarray.