Human neutrophil collagenase splice variant

Abstract

The subject invention is related to human MMP-8alt genes and gene products and their differential expression when comparing a patient with a disease state to a control. A further aspect of the invention concerns compounds which antagonize the biological activity of MMP-8alt protein and methods for identifying these compounds. Another aspect of the present invention concerns pharmaceutical compositions comprising such compounds for the treatment of arthritis, cancer, and disease caused by cellular apoptosis including but not limited to Parkinson's disease, Alzheimer's disease and Huntington's chorea.

What is claimed is:  
     
       1. An isolated polynucleotide comprising a member selected from the group consisting of: 
       (a) a polynucleotide having at least 90% identity to a polynucleotide encoding a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO:2; and  
       (b) a polynucleotide which is complementary to the polynucleotide of (a).  
     
     
       2. The polynucleotide of claim  1  wherein the polynucleotide is DNA. 
     
     
       3. The polynucleotide of claim  2  which encodes the polypeptide comprising an amino acid sequence as set forth in SEQ ID NO:2. 
     
     
       4. The polynucleotide of claim  2  which encodes the polypeptide comprising amino acid 77 to amino acid 238 of SEQ ID NO:2. 
     
     
       5. The polynucleotide of claim  1  wherein the polynucleotide is RNA. 
     
     
       6. The polynucleotide of claim  1  wherein the polynucleotide is genomic DNA. 
     
     
       7. The polynucleotide of claim  1  comprising the sequence as set forth in SEQ ID NO:1. 
     
     
       8. An isolated polynucleotide having at least 95% identity to a polynucleotide comprising nucleotide 174 to nucleotide 2314 of SEQ ID NO:2. 
     
     
       9. An isolated polypeptide comprising an amino acid sequence which is at least 90% identical to the amino acid sequence set forth in SEQ ID NO:2. 
     
     
       10. The polypeptide of claim  9  wherein the polypeptide comprises amino acid 77 to amino acid 238 of SEQ ID NO:2. 
     
     
       11. The polypeptide of claim  9  comprising the amino acid sequence of SEQ ID NO:2. 
     
     
       12. A process for producing a human MMP-8alt polypeptide comprising: 
       (a) transforming a host cell with a vector comprising DNA which upon expression encodes the polypeptide of claim  9 ;  
       (b) culturing the host cell under conditions promoting expression of the polypeptide; and  
       (c) recovering the expressed polypeptide.  
     
     
       13. A human MMP-8alt polypeptide prepared by the process of claim  12 . 
     
     
       14. A process for producing a cell which expresses a polypeptide comprising transforming the cell with a vector comprising DNA which upon expression encodes the polypeptide of claim  9 . 
     
     
       15. A cell prepared by the process of claim  14 . 
     
     
       16. A host cell transformed with a vector comprising DNA which upon expression encodes the polypeptide of claim  9 . 
     
     
       17. An antibody which binds to the polypeptide of claim  9 . 
     
     
       18. A method of producing an antibody comprising: injecting MMP-8alt into a mammal; 
       purifying the antibody produced by the mammal; and  
       recovering the antibody.  
     
     
       19. A method for diagnosing the presence and/or progression of a disease state in a mammal comprising: 
       (a) isolating an MMP-8alt polynucleotide from a body tissue sample derived from the mammal;  
       (b) quantifying the level of expression of the MMP-8alt polynucleotide; and  
       (c) comparing the level of expression determined in step (b) to a control to detect the presence or absence of an over-expression of the polynucleotide.  
     
     
       20. The method of claim  19  wherein the body tissue sample is selected from the group consisting of, urine, blood and synovial fluid. 
     
     
       21. The method of claim  19  wherein the disease state is selected from the group consisting of arthritis, cancer, Parkinson's disease, Alzheimer's disease and Huntington's chorea. 
     
     
       22. The method of claim  19  wherein the polynucleotide is mRNA and the level of mRNA is quantified via RT-PCR. 
     
     
       23. A method for diagnosing the presence and/or progression of a disease state in a mammal comprising: 
       (a) isolating an MMP-8alt polypeptide from a body tissue sample derived from the mammal;  
       (b) quantifying the amount of MMP-8alt polypeptide in the sample; and  
       (c) comparing the amount determined in step (b) to a control to determine the presence or absence of an elevated quantity of the polypeptide.  
     
     
       24. The method of claim  23  wherein the body tissue sample is selected from the group consisting of urine, blood and synovial fluid. 
     
     
       25. The method of claim  23  wherein the disease state is selected from the group consisting of arthritis, cancer, Parkinson's disease, Alzheimer's disease and Huntington's chorea. 
     
     
       26. A method for identifying substances that antagonize or prevent the activity of MMP-8alt protein comprising: 
       (a) contacting MMP-8alt protein with a substance; and  
       (b) determining the ability of the substance to antagonize or prevent the activity of MMP-8alt protein as compared to a control.  
     
     
       27. The method of claim  26  wherein the MMP-8alt protein and substance are combined with a substrate of the MMP-8alt protein under conditions sufficient for the MMP-8alt protein to act on the substrate and the ability of the substance to prevent MMP-8alt from acting on the substrate is determined by quantifying the amount of substrate cleaved by MMP-8alt protein. 
     
     
       28. A substance identified by the method of claim  26 . 
     
     
       29. A method for treating a disease associated with an over-expression of MMP-8alt protein comprising administering a therapeutically effective amount of the substance of claim  28  to a subject in need thereof. 
     
     
       30. A pharmaceutical composition comprising the substance of claim  28  and a pharmaceutically acceptable carrier. 
     
     
       31. A diagnostic kit for detecting a disease state associated with an over-expression of MMP-8alt protein comprising an anti-human MMP-8alt antibody; 
       A diagnostic kit for detecting disease states associated with an over-expression of human MMP-8alt mRNA.