US2014295459A1PendingUtilityA1

Markers for identifying tumor cells, methods and kit thereof

Assignee: ONCOSTEM DIAGNOSTICS MAURITIUS PVT LTDPriority: Mar 27, 2011Filed: Jun 13, 2014Published: Oct 2, 2014
Est. expiryMar 27, 2031(~4.7 yrs left)· nominal 20-yr term from priority
Inventors:Manjiri Bakre
A61P 35/00C07K 14/4748G01N 2800/52G01N 33/57515G01N 33/575G01N 33/5759G01N 33/563C12N 5/0695G01N 33/48A61P 15/00G01N 33/57492
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Claims

Abstract

The present disclosure relates to a combination of biological markers for identification of prognosis of cancer. The present disclosure further relates to a method of identifying the said markers, a method of predicting prognosis and a method of planning personalized treatment for cancer. The present disclosure further relates to a kit/test comprising the antibodies against/other methods of detecting said markers for the said prediction.

Claims

exact text as granted — not AI-modified
We claim: 
     
         1 . A method of prognosis of a subject having breast cancer or suspected of having breast cancer, said method comprising acts of:
 a) collecting biological sample from the subject and identifying expression or absence of receptors selected from a group comprising estrogen receptor, progesterone receptor and Human Epidermal Growth Factor Receptor receptor or any combination thereof in cells of the sample to obtain expression of identified cells;   b) carrying out immunohistochemistry analysis on the cells of step (a) with antibody against a combination of: biological markers consisting of CD44 and P-cadherin; along with at least one biological marker selected from a group consisting of ABCC4, β-catenin, ABCG2, CD133 and Oct-4 or any combination thereof;   c) identifying expression or absence of the receptors and the biological markers in cells of the sample and correlating the identification of the markers with a corresponding outcome based on combination of predictive outcome reference table nos. 11, 12, 13, 14, 15 and 16 for predicting the prognosis of the subject:   
       
         
           
                 
                 
                 
                 
               
                   TABLE 11 
                 
                     
                 
                   MARKER 
                   ER+/PR+ 
                   ER−/PR− 
                   RESULTS 
                 
                     
                 
                   CD44 
                   YES 
                   — 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF CD44 IN 
                 
                     
                     
                     
                   MEMBRANE 
                 
                     
                   YES 
                   — 
                   BAD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF CD44 IN 
                 
                     
                     
                     
                   MEMBRANE 
                 
                     
                     
                     
                   OR 
                 
                     
                     
                     
                   MERE CYTOPLASMIC/NUCLEAR 
                 
                     
                     
                     
                   EXPRESSION 
                 
                   CD44 
                   — 
                   YES 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF CD44 IN 
                 
                     
                     
                     
                   MEMBRANE 
                 
                     
                   — 
                   YES 
                   BAD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF CD44 IN 
                 
                     
                     
                     
                   MEMBRANE 
                 
                     
                     
                     
                   OR 
                 
                     
                     
                     
                   MERE CYTOPLASMIC/NUCLEAR 
                 
                     
                     
                     
                   EXPRESSION 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         
           
                 
                 
                 
                 
               
                   TABLE 12 
                 
                     
                 
                   MARKER 
                   ER+/PR+ 
                   ER−/PR− 
                   RESULTS 
                 
                     
                 
                   ABCG2 
                   — 
                   YES 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF ABCG2 
                 
                     
                     
                     
                   IN NUCLEUS 
                 
                     
                   — 
                   YES 
                   BAD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF ABCG2 
                 
                     
                     
                     
                   IN NUCLEUS 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
               
            
           
         
         
           
                 
                 
                 
                 
               
                   TABLE 13 
                 
                     
                 
                   MARKER 
                   ER+/PR+ 
                   ER−/PR− 
                   RESULTS 
                 
                     
                 
                   ABCC4 
                   YES 
                   — 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF ABCC4 
                 
                     
                     
                     
                   IN MEMBRANE 
                 
                     
                   YES 
                   — 
                   BAD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF ABCC4 
                 
                     
                     
                     
                   IN MEMBRANE 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
               
            
           
         
         
           
                 
                 
                 
                 
               
                   TABLE 14 
                 
                     
                 
                   MARKER 
                   ER+/PR+ 
                   ER−/PR− 
                   RESULTS 
                 
                     
                 
                   CD133 
                   — 
                   YES 
                   GOOD OUTCOME (HER2 −ve) 
                 
                     
                     
                     
                   LOW EXPRESSION OF CD133 
                 
                     
                     
                     
                   IN MEMBRANE 
                 
                     
                   — 
                   YES 
                   BAD OUTCOME (HER2 −ve) 
                 
                     
                     
                     
                   HIGH EXPRESSION OF CD133 
                 
                     
                     
                     
                   IN MEMBRANE 
                 
                   CD133 
                   — 
                   YES 
                   GOOD OUTCOME (HER2 +ve) 
                 
                     
                     
                     
                   HIGH EXPRESSION OF CD133 
                 
                     
                     
                     
                   IN MEMBRANE 
                 
                     
                   — 
                   YES 
                   BAD OUTCOME (HER2 +ve) 
                 
                     
                     
                     
                   LOW EXPRESSION OF CD133 
                 
                     
                     
                     
                   IN MEMBRANE 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         
           
                 
                 
                 
                 
               
                   TABLE 15 
                 
                     
                 
                   MARKER 
                   ER+/PR+ 
                   ER−/PR− 
                   RESULTS 
                 
                     
                 
                   P-CADHERIN 
                   YES 
                   — 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF P- 
                 
                     
                     
                     
                   CADHERIN IN MEMBRANE 
                 
                     
                   YES 
                   — 
                   BAD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF P- 
                 
                     
                     
                     
                   CADHERIN IN MEMBRANE 
                 
                   P-CADHERIN 
                   — 
                   YES 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF P- 
                 
                     
                     
                     
                   CADHERIN IN MEMBRANE 
                 
                     
                   — 
                   YES 
                   BAD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF P- 
                 
                     
                     
                     
                   CADHERIN IN MEMBRANE 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         
           
                 
                 
                 
                 
               
                   TABLE 16 
                 
                     
                 
                   MARKER 
                   ER+/PR+ 
                   ER−/PR− 
                   RESULTS 
                 
                     
                 
                   β-CATENIN 
                   YES 
                   — 
                   GOOD OUTCOME 
                 
                     
                     
                     
                   LOW EXPRESSION OF β- 
                 
                     
                     
                     
                   CATENIN IN MEMBRANE 
                 
                     
                   YES 
                   — 
                   BAD OUTCOME 
                 
                     
                     
                     
                   HIGH EXPRESSION OF β- 
                 
                     
                     
                     
                   CATENIN IN MEMBRANE 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
               
            
           
         
       
       Wherein, the low expression represents percentage intensity of staining of the cells between 0 to 40%, and the high expression represents percentage intensity of staining of the cells between more than 40% and upto 100%. 
     
     
         2 . The method as claimed in  claim 1 , wherein the immunohistochemistry analysis is carried out by conventional method and wherein the identification of markers is carried out by visualizing a colored reaction or fluorescence obtained at completion of the method due to staining of the cells from the sample. 
     
     
         3 . The method as claimed in  claim 1 , wherein the correlating is based on parameters selected from a group comprising percentage of staining, intensity of staining and location of staining or any combination thereof; and wherein the location of the staining is selected from a group comprising cell membrane, cytoplasm, nucleus, and nuclear membrane or any combination thereof. 
     
     
         4 . The method as claimed in  claim 1 , wherein the correlating comprises multiplying the percentage of staining with the intensity of staining to arrive at a predictive score, or wherein the correlating is carried out based on percentage of cells stained in order to predict the prognosis as being good or bad depending on the location of expression of the biological markers. 
     
     
         5 . The combination as claimed in  claim 1 , wherein the combination of biological markers consisting of CD44, ABCC4, P-cadherin and β-catenin prognose breast cancer in a subject having cells expressing estrogen receptor or progesterone receptor or both. 
     
     
         6 . The combination as claimed in  claim 1 , wherein the combination of biological markers consisting of CD44, ABCG2, CD133 and P-cadherin optionally along with Oct-4 prognose breast cancer in a subject having cells not expressing estrogen receptor and progesterone receptor. 
     
     
         7 . The combination as claimed in  claim 1 , wherein the combination of biological markers consisting of CD44, CD133 and P-cadherin prognose breast cancer in a subject having cells expressing Human Epidermal Growth Factor Receptor 2, and not expressing estrogen receptor and progesterone receptor. 
     
     
         8 . A method of treating breast cancer, said method comprising acts of:
 a) identifying a combination of biological markers on tumor cells in a biological sample and predicting prognosis of a subject having breast cancer or suspected of having breast cancer; and   b) based on the prediction, designing a cancer therapy for suppression of the cancer;   
       wherein the combination comprises biological markers consisting of CD44 and P-cadherin; along with at least one biological marker selected from a group consisting of ABCC4, β-catenin, ABCG2, CD133 and Oct-4 or any combination thereof. 
     
     
         9 . A kit for prognosis of a subject having breast cancer or suspected of having cancer, said kit comprising antibody against combination of: biological markers consisting of CD44 and P-cadherin; along with at least one biological marker selected from a group consisting of ABCC4, β-catenin, ABCG2, CD133 and Oct-4 or any combination thereof, optionally along with organic solvent, reagent, secondary antibody, enzyme for performing immunohistochemistry and instruction manual. 
     
     
         10 . The kit as claimed in  claim 9 , wherein the organic solvent is selected from a group comprising alcohol and xylene or a combination thereof. 
     
     
         11 . The kit as claimed in  claim 9 , wherein the combination of biological markers consisting of CD44, ABCC4, P-cadherin and β-catenin prognose breast cancer in the subject having cells expressing estrogen receptor or progesterone receptor or both. 
     
     
         12 . The kit as claimed in  claim 9 , wherein the combination of biological markers consisting of CD44. ABCG2, CD133 and P-cadherin optionally along with Oct-4 prognose breast cancer in the subject having cells not expressing estrogen receptor and progesterone receptor. 
     
     
         13 . The kit as claimed in  claim 9 , wherein the combination of biological markers consisting of CD44, CD133 and P-cadherin prognose breast cancer in the subject having cells expressing Human Epidermal Growth Factor Receptor 2, and not expressing estrogen receptor and progesterone receptor.

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