Gastrointestinal proliferative factor and uses thereof

Abstract

The invention relates to pharmaceutical compositions comprising gastrointestinal proliferative factor (GIPF) polynucleotides and polypeptides. The invention further relates to the therapeutic use of GIPF to prevent or treat conditions or disorders associated with the degeneration of the epithelial mucosa.

1 . A composition comprising a therapeutically effective amount of a GIPF polypeptide, fragment, or analog thereof, and a pharmaceutically acceptable carrier.  
     
     
         2 . A pharmaceutical composition comprising a polypeptide comprising a biologically active fragment of GIPF and a pharmaceutically acceptable carrier.  
     
     
         3 . The pharmaceutical composition of  claim 2  wherein the GIPF is human GIPF.  
     
     
         4 . The pharmaceutical composition of  claim 2  wherein polypeptide comprises a biologically active fragment of the polypeptide of SEQ ID NO:4.  
     
     
         5 . A pharmaceutical composition comprising a polypeptide comprising a polypeptide fragment of SEQ ID NO:4 wherein the polypeptide fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 10, 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, and 178.  
     
     
         6 . The pharmaceutical composition of  claim 1  wherein the polypeptide is glycosylated.  
     
     
         7 . The pharmaceutical composition of  claim 1  wherein the polypeptide is not glycosylated.  
     
     
         8 . The pharmaceutical composition of any of  claim 1  wherein the polypeptide stimulates epithelial cell proliferation.  
     
     
         9 . The pharmaceutical composition of  claim 1  comprising a polypeptide comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:4.  
     
     
         10 . A method of stimulating epithelial cell proliferation in a subject comprising administering to the subject a composition comprising a GIPF polypeptide, fragment or analog thereof and a carrier.  
     
     
         11 . A method of treatment comprising administering to a mammalian subject in need thereof a therapeutically effective amount of a composition comprising a GIPF polypeptide and a pharmaceutically acceptable carrier.  
     
     
         12 . A method of treating mucositis, inflammatory bowel disease, or short bowel syndrome comprising administering to a mammalian subject in need thereof a therapeutically effective amount of a composition comprising a GIPF polypeptide and a pharmaceutically acceptable carrier.  
     
     
         13 . The method for stimulating epithelial cell proliferation in the gastrointestinal tract of a patient, the method comprising administering a therapeutically effective amount of a composition comprising a GIPF polypeptide and a pharmaceutically acceptable carrier.  
     
     
         14 . The method of  claim 13  wherein epithelial cell proliferation in the esophagus is stimulated.  
     
     
         15 . The method of  claim 13  wherein epithelial cell proliferation in the small intestine is stimulated.  
     
     
         16 . The method of  claim 13  wherein epithelial cell proliferation in the large intestine is stimulated.  
     
     
         17 . The method of  claim 13  wherein epithelial cell proliferation in the stomach is stimulated.  
     
     
         18 . A method for treating a patient at risk for damage to epithelial cells lining at least a portion of the gastrointestinal tract, the method comprising administering a therapeutically effective amount of a composition comprising a GIPF polypeptide and a pharmaceutically acceptable carrier.  
     
     
         19 . The method of  claim 18  wherein the patient has undergone or will undergo radiation therapy.  
     
     
         20 . The method of  claim 18  wherein the patient has undergone or will undergo chemotherapy.  
     
     
         21 . A method for treating a patient that has undergone radiation therapy or chemotherapy comprising administering a therapeutically effective amount of a composition comprising a GIPF polypeptide and a pharmaceutically acceptable carrier.  
     
     
         22 . An adenoviral vector comprising a gene encoding GIPF operably associated with an expression control sequence.  
     
     
         23 . A pharmaceutical composition comprising the adenoviral vector of  claim 22 .  
     
     
         24 . A method of stimulating epithelial cell proliferation in a subject comprising administering to said subject the pharmaceutical composition of  claim 23 .  
     
     
         25 . A transgene construct comprising a nucleic acid encoding a GIPF protein, wherein said nucleic acid is operably linked to transcriptional regulatory sequences directing its expression in B-cells.  
     
     
         26 . The transgene construct of  claim 25  comprising a B-cell specific promoter.  
     
     
         27 . A transgenic mouse that produces in its B-cells cells detectable levels of a native human GIPF protein, wherein said transgenic mouse has stably integrated into its genome a nucleic acid sequence encoding a GIPF protein, operably linked to transcriptional regulatory sequences directing its expression to B-cells.  
     
     
         28 . The transgenic mouse of  claim 27 , wherein said transcriptional regulatory sequences comprise a B-cell promoter.  
     
     
         29 . A method of identifying a drug candidate for the treatment of mucositis, inflammatory bowel disease or short bowel syndrome comprising 
 (a) administering a test compound to a transgenic mouse that expresses in its B-cells a recombinant GIPF polypeptide and exhibits increased intestinal epithelial cell proliferation compared to an otherwise identical mouse not expressing a recombinant GIPF polypeptide intestinal epithelial cell proliferation; and    (b) evaluating the effect of said test compound on intestinal epithelial cell proliferation, wherein an increase in intestinal cell proliferation identifies the test compound as a drug candidate for the treatment of mucositis, inflammatory bowel disease or short bowel syndrome.    
     
     
         30 . The method of  claim 29 , wherein the intestinal epithelial cell is a crypt cell.  
     
     
         31 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 9, 11, 13, 15, 17, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, and 104.  
     
     
         32 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, and 105.  
     
     
         33 . A method for producing a polypeptide comprising the amino acid sequence of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, or 105, the method comprising: 
 (a) culturing an isolated cell comprising a nucleic acid molecule encoding a polypeptide of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, or 105 in culture medium under conditions suitable for expressing the polypeptide; and    (b) purifying the polypeptide from the cell or the culture medium.    
     
     
         34 . A method for producing a pharmaceutical composition comprising a polypeptide comprising the amino acid sequence of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105 or 178, the method comprising 
 (a) culturing an isolated cell comprising a nucleic acid molecule encoding a polypeptide of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, or 178 in culture medium under conditions suitable for expressing the polypeptide;    (b) purifying the polypeptide from the cell or the culture medium; and    (c) combining the purified polypeptide with a pharmaceutically acceptable carrier.    
     
     
         35 . A method for producing a pharmaceutical composition comprising a polypeptide comprising the amino acid sequence of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105 or 178, the method comprising 
 (a) synthesizing comprising a polypeptide comprising the amino acid sequence of SEQ ID NO: 12, 14, 16, 18, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105 or 178;    (b) purifying the polypeptide; and    (c) combining the purified polypeptide with a pharmaceutically acceptable carrier.    
     
     
         36 . A expression vector construct comprising a nucleic acid encoding a GIPF protein, wherein the nucleic acid is operably linked to transcriptional regulatory sequences directing expression of the GIPF protein in intestinal epithelial cells.  
     
     
         37 . A transgenic mouse that produces in its intestinal epithelial cells detectable levels of a GIPF protein, wherein said transgenic mouse has stably integrated into its genome a nucleic acid sequence encoding a GIPF protein, wherein the nucleic acid sequence encoding the GIPF protein is operably linked to transcriptional regulatory sequences directing expression of the GIPF protein in intestinal epithelial cells.  
     
     
         38 . A expression vector construct comprising a nucleic acid encoding a GIPF protein and a Wnt3a protein, wherein the nucleic acid is operably linked to transcriptional regulatory sequences directing its expression in intestinal epithelial cells.  
     
     
         39 . A transgenic mouse that produces in its intestinal epithelial cells detectable levels of a native GIPF and Wnt3a protein, wherein said transgenic mouse has stably integrated into its genome a nucleic acid sequence encoding a GIPF protein, operably linked to transcriptional regulatory sequences directing expression of the GIPF protein in intestinal epithelial cells.  
     
     
         40 . The method of  claim 13  wherein epithelial cell proliferation in the oral cavity is stimulated.