Diagnosis and treatment of malignant neoplasms

Abstract

The invention features a method for diagnosing a malignant neoplasm in a mammal by contacting a bodily fluid from the mammal with an antibody which binds to an human aspartyl (asparaginyl) beta-hydroxylase (HAAH) polypeptide and methods of treating malignant neoplasms by inhibiting HAAH.

What is claimed is:  
     
         1 . A method for diagnosing a malignant neoplasm in a mammal, comprising contacting a bodily fluid from said mammal with an antibody which binds to an human aspartyl (asparaginyl) beta-hydroxylase (HAAH) polypeptide under conditions sufficient to form an antigen-antibody complex and detecting the antigen-antibody complex.  
     
     
         2 . The method of  claim 1 , wherein said neoplasm is derived from endodermal tissue.  
     
     
         3 . The method of  claim 1 , wherein said neoplasm is selected from the group consisting of colon cancer, breast cancer, pancreatic cancer, liver cancer, and cancer of the bile ducts.  
     
     
         4 . The method of  claim 1 , wherein said neoplasm is a cancer of the central nervous system (CNS).  
     
     
         5 . The method of  claim 1 , wherein said bodily fluid is selected from the group consisting of a CNS-derived bodily fluid, blood, serum, urine, saliva, sputum, lung effusion, and ascites fluid.  
     
     
         6 . The method of  claim 1 , wherein said antibody is a monoclonal antibody.  
     
     
         7 . The method of  claim 6 , wherein said monoclonal antibody is FB50.  
     
     
         8 . The method of  claim 6 , wherein said monoclonal antibody is selected from the group consisting of 5C7, 5E9, 19B, 48A, 74A, 78A, 86A.  
     
     
         9 . A method for prognosis of a malignant neoplasm of a mammal, comprising 
 (a) contacting a bodily fluid from said mammal with an antibody which binds to an HAAH polypeptide under conditions sufficient to form an antigen-antibody complex and detecting the antigen-antibody complex;    (b) quantitating the amount of complex to determine the level of HAAH in said fluid; and    (c) comparing the level of HAAH in said fluid with a normal control level of HAAH, wherein increasing levels of HAAH over time indicates an adverse prognosis.    
     
     
         10 . A method of inhibiting tumor growth in a mammal comprising administering to said mammal a compound which inhibits expression of HAAH.  
     
     
         11 . The method of  claim 10 , wherein said compound is a HAAH antisense nucleic acid.  
     
     
         12 . The method of  claim 10 , wherein said compound is a ribozyme.  
     
     
         13 . The method of  claim 10 , wherein said tumor is derived from endodermal tissue.  
     
     
         14 . The method of  claim 10 , wherein said tumor is selected from the group consisting of colon cancer, breast cancer, pancreatic cancer, liver cancer, and cancer of the bile ducts.  
     
     
         15 . The method of  claim 10 , wherein said tumor is a CNS tumor.  
     
     
         16 . A method of inhibiting tumor growth in a mammal comprising administering to said mammal a compound which inhibits an enzymatic activity of HAAH.  
     
     
         17 . The method of  claim 16 , wherein said enzymatic activity is hydroxylase activity.  
     
     
         18 . The method of  claim 16 , wherein said compound is a dominant negative mutant of HAAH.  
     
     
         19 . The method of  claim 18 , wherein said dominant negative mutant HAAH comprises a mutation in a catalytic domain of HAAH.  
     
     
         20 . The method of  claim 16 , wherein said compound is an HAAH-specific intrabody.  
     
     
         21 . The method of  claim 16 , wherein said compound is L-mimosine.  
     
     
         22 . The method of  claim 16 , wherein said compound is a hydroxypyridone.  
     
     
         23 . A method of inhibiting tumor growth in a mammal comprising administering to said mammal a compound which inhibits signal transduction through the IRS signal transduction pathway.  
     
     
         24 . The method of  claim 23 , wherein said compound inhibits IRS phosphorylation.  
     
     
         25 . The method of  claim 23 , wherein said compound inhibits binding of Fos or Jun to an HAAH promoter sequence.  
     
     
         26 . A method of inhibiting tumor growth in a mammal comprising administering to said mammal a compound which inhibits HAAH hydroxylation of a NOTCH polypeptide.  
     
     
         27 . The method of  claim 26 , wherein said compound inhibits hydroxylation of an EGF-like repeat sequence in a NOTCH polypeptide.  
     
     
         28 . A method of killing a tumor cell comprising contacting said tumor cell with cytotoxic agent linked to an HAAH-specific antibody.  
     
     
         29 . A monoclonal antibody that binds to an epitope of HAAH.  
     
     
         30 . The antibody of  claim 29 , wherein said epitope is within a catalytic site of HAAH.  
     
     
         31 . The antibody of  claim 29 , wherein said monoclonal antibody is selected from the group consisting of 5C7, 5E9, 19B, 48A, 74A, 78A, 86A.  
     
     
         32 . The antibody of  claim 29 , wherein said monoclonal antibody is selected from the group consisting of HA238A, HA221, HA239, HA241, HA329, or HA355.  
     
     
         33 . A composition comprising a monoclonal antibody that binds to an epitope of HAAH linked to a cytotoxic agent, wherein said composition preferentially kills tumor cells compared to non-tumor cells.  
     
     
         34 . A kit for diagnosis of a tumor in a mammal, comprising the antibody of  claim 29 .  
     
     
         35 . The kit of  claim 34 , wherein said antibody is immobilized on a solid phase.  
     
     
         36 . The kit of  claim 35 , wherein said solid phase is selected from a group consisting of an assay plate, an assay well, a nitrocellulose membrane, a bead, a dipstick, and a component of an elution column.  
     
     
         37 . A method of determining whether a candidate compound inhibits HAAH enzymatic activity, comprising 
 (a) providing a HAAH polypeptide;    (b) providing a polypeptide comprising an EGF—like domain;    (c) contacting said HAAH polypeptide or said NOTCH polypeptide with said candidate compound;    (d) determining hydroxylation of said polypeptide of step (b), wherein a decrease in hydroxylation in the presence of said candidate compound compared to that in the absence of said compound indicates that said compound. inhibits HAAH enzymatic activity.    
     
     
         38 . A method of determining whether a candidate compound inhibits HAAH activation of NOTCH, comprising 
 (a) providing a cell expressing HAAH;    (b) contacting said cell with a candidate compound; and    (c) measuring translocation of activated NOTCH to the nucleus of said cell, wherein a decrease in translocation in the presence of said compound compared to that in the absence of said compound indicates that said compound HAAH activation of NOTCH.