Methods and means for treatment of osteoarthritis

Abstract

The present invention relates to in vivo and in vitro methods, agents and compound screening assays for inducing anabolic stimulation of chondrocytes, including cartilage formation enhancing pharmaceutical compositions, and the use thereof in treating and/or preventing a disease involving a systemic or local decrease in mean cartilage thickness in a subject.

1 . Method for identifying a compound that induces chondrocyte anabolic stimulation, comprising:
 contacting a compound with a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 55-82 and 198-391, and 480; and   measuring a compound-polypeptide property related to the anabolic stimulation of chondrocytes.   
     
     
         2 . The method according to  claim 1 , wherein said polypeptide is in an in vitro cell-free preparation. 
     
     
         3 . The method according to  claim 2 , wherein said polypeptide is present in a mammalian cell. 
     
     
         4 . The method of  claim 1 , wherein said property is a binding affinity of said compound to said polypeptide. 
     
     
         5 . The method of  claim 3 , wherein said property is activation of a biological pathway producing a biochemical marker indicative of the anabolic stimulation of chondrocytes. 
     
     
         6 . The method of  claim 5  wherein said indicator is selected from the group consisting of collagen type II, alpha-1 (col2a1) and aggrecan. 
     
     
         7 . The method of  claim 6  wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 55-82. 
     
     
         8 . The method according to  claim 1 , wherein said compound is selected from the group consisting of compounds of a commercially available screening library and compounds having binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 55-82 and 198-391, and 480. 
     
     
         9 . The method according to  claim 2 , wherein said compound is a peptide in a phage display library or an antibody fragment library. 
     
     
         10 . An agent for inducing the anabolic stimulation of chondrocytes, selected from the group consisting of an antisense polynucleotide, a ribozyme, and a small interfering RNA (siRNA), wherein said agent comprises a nucleic acid sequence complementary to, or engineered from, a naturally-occurring polynucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 55-82 and 198-391, and 480. 
     
     
         11 . The agent according to  claim 10 , wherein polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 55-82. 
     
     
         12 . The agent according to  claim 11 , wherein a vector in a mammalian cell expresses said agent. 
     
     
         13 . The agent according to  claim 12 , wherein said vector is an adenoviral, retroviral, adeno-associated viral, lentiviral, a herpes simplex viral or a sendaiviral vector. 
     
     
         14 . The agent according to  claim 10 , wherein said antisense polynucleotide and said siRNA comprise an antisense strand of 17-25 nucleotides complementary to a sense strand, wherein said sense strand is selected from 17-25 continuous nucleotides of a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 55-82. 
     
     
         15 . The agent according to  claim 14 , wherein said siRNA further comprises said sense strand. 
     
     
         16 . The agent according to  claim 15 , wherein said sense strand is selected from 17-25 continuous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-28 and 479. 
     
     
         17 . The agent according to  claim 16 , wherein said siRNA further comprises a loop region connecting said sense and said antisense strand. 
     
     
         18 . The agent according to  claim 17 , wherein said loop region comprises a nucleic acid sequence defined of SEQ ID NO: 83. 
     
     
         19 . The agent according to  claim 18 , wherein said agent is an antisense polynucleotide, ribozyme, or siRNA comprising a nucleic acid sequence complementary to a polynucleotide selected from the group consisting of SEQ ID NO: 84-197 and 410. 
     
     
         20 . A chondrocyte anabolic stimulation enhancing pharmaceutical composition comprising a therapeutically effective amount of an agent of  claim 10  in admixture with a pharmaceutically acceptable carrier. 
     
     
         21 - 32 . (canceled)