Novel tissue protective erythropoietin receptor (nepor) and methods of use

Abstract

Methods of determining whether a patient is suitable for erythropoietin (EPO) therapy, including (A) isolating a tissue sample from said patient; (B) determining the level of expression of EPH-B4 in said sample; and (C) correlating a presence of EPH-B4 expression to a negative physiological response to EPO therapy). In addition, methods of enhancing the effectiveness of EPO therapy in a patient by administering to said patient, in conjunction with EPO therapy, an siRNA specific for EPH-B4.

1 .- 21 . (canceled) 
     
     
         22 . A method of determining whether a patient is suitable for erythropoietin (EPO) therapy, comprising
 (A) isolating a tissue sample from said patient;   (B) determining the level of expression of EPH-B4 in said sample; and   (C) correlating a presence of EPH-B4 expression to a negative physiological response to EPO therapy.   
     
     
         23 . The method of  claim 22 , wherein the level of expression is determined by measuring the amount of EPH-B4 protein in said sample. 
     
     
         24 . The method of  claim 23 , wherein the presence of EPH-B4 expression is defined by the percentage of cells in said sample showing detectable levels of EPH-B4 protein and the concentration of EPH-B4 protein in said cells. 
     
     
         25 . The method of  claim 23 , wherein the presence of EPH-B4 expression is defined by the formula
   P×C
   
       wherein P is the percentage of cells in said sample showing detectable levels of EPH-B4 protein and C is the relative concentration of EPH-B4 protein in said cells,
 wherein a score of 0, 1, 2, 3 or 4 is assigned to a sample comprising a percentage of cells showing detectable levels of EPH-B4 protein of, respectively, 0%, <25%, 25-50%, 50-75% and 75-100%, 
 wherein a score of 1, 2 or 3 is assigned to relative concentrations of EPH-B4 protein of, respectively, weak, moderate and heavy, and 
 wherein a resulting product of >3 denotes EPH-B4 expression in the sample. 
 
     
     
         26 . The method of  claim 23 , wherein the level of expression is determined by a technique selected from the group consisting of immunoassay, chemiluminescent assay, nephelometric assay, turbidimetric assay, bioassay and reporter-assay. 
     
     
         27 . The method of  claim 23 , wherein the level of expression is determined by an immunoassay selected from the group consisting of enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, enzyme immunoassay (EIA), radioimmunoassay (RIA), fluorescent immunoassay, Western blot, competitive immunoassay, noncompetitive immunoassay, homogeneous immunoassay and heterogeneous immunoassay. 
     
     
         28 . The method of  claim 23 , wherein the level of expression is determined by ELISA. 
     
     
         29 . The method of  claim 23 , wherein said level of expression is determined by immunohistochemistry. 
     
     
         30 . The method of  claim 22 , wherein the level of expression is determined by measuring the amount of EPH-B4 mRNA in said sample. 
     
     
         31 . The method of  claim 29 , wherein the level of expression is determined by a technique selected from the group consisting of PCR, QPCR, R-PCR, gene expression microarray analysis, northern-blot analysis, reverse transcription and amplification, zymography, ligase-chain-reaction, NASBA, RNase Protection Assay (RPA), capillary electrophoresis with laser induced fluorescence (CE-LIF), and RT-PCR. 
     
     
         32 . The method of  claim 22 , wherein the level of expression is determined by RT-PCR. 
     
     
         33 . The method of  claim 22 , wherein the tissue sample is selected from cancerous tissue or circulating cells derived from same. 
     
     
         34 . The method of  claim 22 , wherein the tissue sample is selected from blood, lymph, urine or cerebral fluid. 
     
     
         35 . The method of  claim 22 , further comprising determining the level of expression in said sample of at least one of Ephrin A1 or EPOR. 
     
     
         36 . The method of  claim 22 , further comprising determining the level of expression of Ephrin A1 in said sample. 
     
     
         37 . The method of  claim 22 , wherein said patient is a cancer patient considering EPO therapy. 
     
     
         38 . The method of  claim 37 , wherein said negative physiological response includes the proliferation of cancer cells of said patient. 
     
     
         39 . A method of enhancing the effectiveness of EPO therapy in a patient, comprising administering to said patient, in conjunction with EPO therapy, an siRNA specific for EPH-B4. 
     
     
         40 . The method of  claim 39 , wherein the siRNA is selected from the group of nucleic acid duplexes consisting of SEQ ID NO: 242 and SEQ ID NO: 243; SEQ ID NO: 244 and SEQ ID NO: 245; SEQ ID NO: 246 and SEQ ID NO: 247; SEQ ID NO: 248 and SEQ ID NO: 249; SEQ ID NO: 250 and SEQ ID NO: 251; SEQ ID NO: 252 and SEQ ID NO: 253; SEQ ID NO: 254 and SEQ ID NO: 255; SEQ ID NO: 256 and SEQ ID NO: 257; SEQ ID NO: 258 and SEQ ID NO: 259; SEQ ID NO: 260 and SEQ ID NO: 261; and SEQ ID NO: 266 and SEQ ID NO: 267. 
     
     
         41 . The method of  claim 39 , wherein the siRNA is a duplex of SEQ ID NO: 219 and SEQ ID NO: 220.