Human FGF-21 gene and gene expression products

Abstract

This invention relates to human fibroblast growth factor (hFGF-21), and to variants thereof and to polynucleotides encoding FGF-21. The invention also relates to diagnostic and therapeutic agents related to the polynucleotides and proteins, including probes and antibodies, and to methods of treating liver disease such as cirrhosis and cancer, methods of treating conditions related to thymic function, and methods of treating conditions of the testis. The invention also relates to mouse fibroblast growth factor (mFGF-21), and to variants thereof and polynucleotides encoding mFGF-21.

We claim:  
     
         1 . An isolated nucleic acid molecule comprising a polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding amino acids from about 1 to about 209 of SEQ ID NO:4;    (b) a polynucleotide encoding amino acids from about 2 to about 209 of SEQ ID NO:4;    (c) a polynucleotide encoding amino acids from about 1 to about 177 of SEQ ID NO:4;    (d) a polynucleotide encoding amino acids from about 40 to about 209 of SEQ ID NO:4;    (e) a polynucleotide encoding amino acids from about 40 to about 177 of SEQ ID NO:4;    (f) the polynucleotide complement of (a), (b), (c), (d) or (e); and    (g) a polynucleotide at least 90% identical to the polynucleotide of (a), (b), (c), (d) or (e).    
     
     
         2 . An isolated nucleic acid molecule which comprises 20-600 contiguous nucleotides from the coding region of SEQ ID NO:3.  
     
     
         3 . The isolated nucleic acid molecule of  claim 2 , which comprises 60-400 contiguous nucleotides from the coding region of SEQ ID NO:3.  
     
     
         4 . The isolated nucleic acid molecule of  claim 3 , which comprises 200-300 contiguous nucleotides from the coding region of SEQ ID NO:3.  
     
     
         5 . An isolated nucleic acid molecule comprising a polynucleotide encoding a polypeptide wherein, except for at least one conservative amino acid substitution, said polypeptide has an amino acid sequence selected from the group consisting of: 
 (a) amino acids from about 1 to about 209 of SEQ ID NO:4;    (b) amino acids from about 2 to about 209 of SEQ ID NO:4;    (c) amino acids from about 1 to about 177 of SEQ ID NO:4;    (d) amino acids from about 40 to about 209 of SEQ ID NO:4; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:4.    
     
     
         6 . The isolated nucleic acid molecule of  claim 1 , which is DNA.  
     
     
         7 . A method of making a recombinant vector comprising inserting a nucleic acid molecule of  claim 1  into a vector in operable linkage to a promoter.  
     
     
         8 . A recombinant vector produced by the method of  claim 7 .  
     
     
         9 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 8  into a host cell.  
     
     
         10 . A recombinant host cell produced by the method of  claim 9 .  
     
     
         11 . A recombinant method of producing a polypeptide, comprising culturing the recombinant host cell of  claim 10  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         12 . An isolated polypeptide comprising amino acids at least 95% identical to amino acids selected from the group consisting of: 
 (a) amino acids from about 1 to about 209 of SEQ ID NO:4;    (b) amino acids from about 2 to about 209 of SEQ ID NO:4;    (c) amino acids from about 1 to about 177 of SEQ ID NO:4;    (d) amino acids from about 40 to about 209 of SEQ ID NO:4; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:4.    
     
     
         13 . An isolated polypeptide wherein, except for at least one conservative amino acid substitution, said polypeptide has an amino acid sequence selected from the group consisting of: 
 (a) amino acids from about 1 to about 209 of SEQ ID NO:4;    (b) amino acids from about 2 to about 209 of SEQ ID NO:4;    (c) amino acids from about 1 to about 177 of SEQ ID NO:4;    (d) amino acids from about 40 to about 209 of SEQ ID NO:4; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:4.    
     
     
         14 . An isolated polypeptide comprising amino acids selected from the group consisting of: 
 (a) amino acids from about 1 to about 209 of SEQ ID NO:4;    (b) amino acids from about 2 to about 209 of SEQ ID NO:4;    (c) amino acids from about 1 to about 177 of SEQ ID NO:4;    (d) amino acids from about 40 to about 209 of SEQ ID NO:4; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:4.    
     
     
         15 . An epitope-bearing portion of the polypeptide of SEQ ID NO:4.  
     
     
         16 . The epitope-bearing portion of  claim 15 , which comprises between 10 and 50 contiguous amino acids of SEQ ID NO:4.  
     
     
         17 . The epitope-bearing portion of  claim 15 , which comprises amino acids RQRYLYTDDAQQTEAH (SEQ ID NO:7).  
     
     
         18 . The epitope-bearing portion of  claim 15 , which comprises amino acids HLPGNKSPHRDPAPR (SEQ ID NO:8).  
     
     
         19 . An isolated antibody that binds specifically to the polypeptide of  claim 12 .  
     
     
         20 . An isolated antibody that binds specifically to the polypeptide of  claim 13 .  
     
     
         21 . An isolated antibody that binds specifically to the polypeptide of  claim 14 .  
     
     
         22 . A pharmaceutical composition comprising the polypeptide of  claim 12 , in combination with a pharmaceutically acceptable carrier.  
     
     
         23 . A method for providing trophic support for cells in a patient in need thereof, the method comprising administering to the patient a composition comprising a polynucleotide encoding the polypeptide of SEQ ID NO:4.  
     
     
         24 . The method of  claim 23  wherein said polynucleotide is administered by implanting cells which express said polynucleotide into the patient, wherein said cells express FGF-21 polypeptide in the patient.  
     
     
         25 . The method of  claim 23  wherein the implanted cells are encapsulated in a semipermeable membrane.  
     
     
         26 . The method of  claim 23  wherein the patient suffers from a condition characterized by inadequate numbers of hepatic cells.  
     
     
         27 . The method of  claim 23  wherein the condition is cirrhosis of the liver.  
     
     
         28 . The method of  claim 23  wherein said patient suffers from a condition characterized by inadequate function or number of testicular cells.  
     
     
         29 . The method of  claim 28  wherein said condition is at least one condition selected from the group consisting of infertility, impotence, and testicular cancer.  
     
     
         30 . The method of  claim 23  wherein the patient suffers from a condition characterized by inadequate function of the thymus.  
     
     
         31 . The method of  claim 30  wherein said condition is at least one condition selected from the group consisting of leukemia, lymphoma, autoimmune disease, proliferative disorder of the thymus, and differentiation disorder of the thymus.  
     
     
         32 . A method for providing trophic support for cells in a patient in need thereof, the method comprising administering to the patient a composition comprising a polypeptide of SEQ ID NO:4.  
     
     
         33 . The method of  claim 28  wherein the patient suffers from a condition characterized by inadequate numbers of hepatic cells.  
     
     
         34 . The method of  claim 29  wherein the condition is cirrhosis of the liver.  
     
     
         35 . A method of alleviating a disease condition in the liver of a human patient wherein said disease condition is alleviated by at least one method selected from the group consisting of slowing degeneration of, restoring function of, and increasing the number of, functional hepatic cells in said human patient, said method comprising administering to said patient a pharmaceutically effective composition comprising a polypeptide having the amino acid sequence of SEQ ID NO:4.  
     
     
         36 . A method of alleviating a disease condition in the thymus of a human patient wherein said disease condition is alleviated by at least one method selected from the group consisting of preventing degeneration of, slowing degeneration of, increasing the number of, functional thymic cells in said human patient, said method comprising administering to said patient a pharmaceutically effective composition comprising a polypeptide having the amino acid sequence of SEQ ID NO:4.  
     
     
         37 . A method of alleviating a disease condition in the testis of a human patient wherein said disease condition is alleviated by at least one method selected from the group consisting of preventing degeneration of, slowing degeneration of, and increasing the number of, functional testicular cells in said human patient, said method comprising administering to said patient a pharmaceutically effective composition comprising a polypeptide having the amino acid sequence of SEQ ID NO:4.  
     
     
         38 . A kit for detecting the presence of mRNA encoding FGF-21 in a sample from a patient, said kit comprising a polynucleotide having at least 20 contiguous nucleotides of the polynucleotide of  claim 3 , packaged in a container.  
     
     
         39 . The kit according to  claim 38  wherein the polynucleotide encodes at least six contiguous amino acids of SEQ ID NO:4.  
     
     
         40 . A kit for detecting the presence of FGF-21 polypeptide in a sample from a patient, said kit comprising an antibody according to  claim 19 , packaged in a container.  
     
     
         41 . An isolated nucleic acid molecule comprising a polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding amino acids from about 1 to about 210 of SEQ ID NO:2;    (b) a polynucleotide encoding amino acids from about 2 to about 21 of SEQ ID NO:2;    (c) a polynucleotide encoding amino acids from about 1 to about 177 of SEQ ID NO:2;    (d) a polynucleotide encoding amino acids from about 40 to about 210 of SEQ ID NO:2;    (e) a polynucleotide encoding amino acids from about 40 to about 177 of SEQ ID NO:2;    (f) the polynucleotide complement of (a), (b), (c), (d) or (e); and    (h) a polynucleotide at least 90% identical to the polynucleotide of (a), (b), (c), (d) or (e).    
     
     
         42 . An isolated nucleic acid molecule which comprises 20-600 contiguous nucleotides from the coding region of SEQ ID NO:1.  
     
     
         43 . The isolated nucleic acid molecule of  claim 42 , which comprises 60-400 contiguous nucleotides from the coding region of SEQ ID NO:1.  
     
     
         44 . The isolated nucleic acid molecule of  claim 43 , which comprises 200-300 contiguous nucleotides from the coding region of SEQ ID NO:1.  
     
     
         45 . An isolated nucleic acid molecule comprising a polynucleotide encoding a polypeptide wherein, except for at least one conservative amino acid substitution, said polypeptide has an amino acid sequence selected from the group consisting of: 
 (a) amino acids from about 1 to about 210 of SEQ ID NO:2;    (b) amino acids from about 2 to about 210 of SEQ ID NO:2;    (c) amino acids from about 1 to about 177 of SEQ ID NO:2;    (d) amino acids from about 40 to about 210 of SEQ ID NO:2; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:4.    
     
     
         46 . The isolated nucleic acid molecule of  claim 41 , which is DNA.  
     
     
         47 . A method of making a recombinant vector comprising inserting a nucleic acid molecule of  claim 41  into a vector in operable linkage to a promoter.  
     
     
         48 . A recombinant vector produced by the method of  claim 47 .  
     
     
         49 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 48  into a host cell.  
     
     
         50 . A recombinant host cell produced by the method of  claim 49 .  
     
     
         51 . A recombinant method of producing a polypeptide, comprising culturing the recombinant host cell of  claim 50  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         52 . An isolated polypeptide comprising amino acids at least 95% identical to amino acids selected from the group consisting of: 
 (a) amino acids from about 1 to about 210 of SEQ ID NO:2;    (b) amino acids from about 2 to about 210 of SEQ ID NO:2;    (c) amino acids from about 1 to about 177 of SEQ ID NO:2;    (d) amino acids from about 40 to about 210 of SEQ ID NO:2; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:2.    
     
     
         53 . An isolated polypeptide wherein, except for at least one conservative amino acid substitution, said polypeptide has an amino acid sequence selected from the group consisting of: 
 (a) amino acids from about 1 to about 210 of SEQ ID NO:2;    (b) amino acids from about 2 to about 210 of SEQ ID NO:2;    (c) amino acids from about 1 to about 177 of SEQ ID NO:2;    (d) amino acids from about 40 to about 210 of SEQ ID NO:2; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:2.    
     
     
         54 . An isolated polypeptide comprising amino acids selected from the group consisting of: 
 (a) amino acids from about 1 to about 210 of SEQ ID NO:2;    (b) amino acids from about 2 to about 210 of SEQ ID NO:2;    (c) amino acids from about 1 to about 177 of SEQ ID NO:2;    (d) amino acids from about 40 to about 210 of SEQ ID NO:2; and    (e) amino acids from about 40 to about 177 of SEQ ID NO:2.    
     
     
         55 . An epitope-bearing portion of the polypeptide of SEQ ID NO:2.  
     
     
         56 . The epitope-bearing portion of  claim 55 , which comprises between 10 and 50 contiguous amino acids of SEQ ID NO:2.  
     
     
         57 . An isolated antibody that binds specifically to the polypeptide of  claim 52 .  
     
     
         58 . An isolated antibody that binds specifically to the polypeptide of  claim 53 .  
     
     
         59 . An isolated antibody that binds specifically to the polypeptide of  claim 54.