Antibody recognizing human leukemia inhibitory factor (lif) and use of anti-lif antibodies in the treatment of diseases associated with unwanted cell proliferation

Abstract

The invention relates to antibodies directed against human Leukemia Inhibitory Factor (LIF) and to a hybridoma cell line producing said antibodies. The invention also relates to a method for blocking/inhibiting the proliferation of stem cells, and to an in vitro method for the diagnosis of diseases associated with unwanted cell proliferation in a subject or for determining the predisposition of a subject to suffer from said disease associated with unwanted cell proliferation, or for prognosis of average life expectancy of a subject suffering from said disease. The therapeutic potential of said antibodies is based on observing that the inhibition of LIF can be used in therapeutic compositions for the treatment of diseases associated with unwanted proliferation.

1 . A method for the treatment of cancer comprising administering a monoclonal antibody or a fragment thereof, which binds full length human LIF, but does not bind a LIF fragment corresponding to amino acids 1 to 160, wherein the full length human LIF is the precursor to human LIF isoform 1. 
     
     
         2 . The method of  claim 1 , wherein the cancer is characterized by
 a. high LIF levels or   b. a cell population expressing high levels of CD44 and Id1.   
     
     
         3 . The method of  claim 1  wherein the antibody binds an epitope of human LIF comprised in the region corresponding to amino acids 160 to 202 of human LIF. 
     
     
         4 . The method of  claim 1  wherein said antibody binds an epitope comprised in the regions selected from the following: a region corresponding to amino acids 160 to 180, a region corresponding to amino acids 170 to 190, a region corresponding to amino acids 180 to 200, a region corresponding to amino acids 182 to 202 of human LIF. 
     
     
         5 . The method of  claim 1  wherein the antibody is competitively inhibited in its binding to human LIF by the monoclonal antibody produced by a hybridoma deposited on Apr. 1 2010 by Vall d'Hebron Institute of Oncology at the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM ACC3054). 
     
     
         6 . The method of  claim 1  wherein the monoclonal antibody is of the IgG 1 isotype. 
     
     
         7 . The method of  claim 1  wherein the antibody is a humanized antibody. 
     
     
         8 . The method of  claim 1  wherein the monoclonal antibody is produced by the hybridoma cell line DSM ACC3054, deposited on Apr. 1 2010 by Vall d'Hebron Institute of Oncology at the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH. 
     
     
         9 . The method of  claim 1  wherein the monoclonal antibody or fragment thereof acts through the inhibition of the self-regeneration of tumor stem cells. 
     
     
         10 . The method of  claim 1  wherein the antibody or fragment thereof is capable of reducing the cell population characterized by high levels of CD 44 and Id1. 
     
     
         11 . The method of  claim 1 , wherein the cancer is one of the following: glioma, pre-B cell acute lymphoblastic leukemia, acute myeloid leukemia, lung adenocarcinoma, prostate adenocarcinoma, colorectal carcinoma, bladder cancer, breast ductal cancer or breast carcinoma. 
     
     
         12 . The method of  claim 2 , wherein the cancer is one of the following: glioma, pre-B cell acute lymphoblastic leukemia, acute myeloid leukemia, lung adenocarcinoma, prostate adenocarcinoma, colorectal carcinoma, bladder cancer, breast ductal cancer or breast carcinoma. 
     
     
         13 . An in vitro method for designing a customized therapy for a patient suffering from a disease associated with increased LIF levels comprising:
 (a) quantifying the expression levels of LIF in said patient, and   (b) comparing said expression levels with control levels,   wherein if the expression levels of LIF in said patient are greater than the control values, then a monoclonal antibody or a fragment thereof is administered to said patient, wherein the monoclonal antibody or a fragment thereof, binds full length human LIF, but does not bind a LIF fragment corresponding to amino acids 1 to 160, wherein the full length human LIF is the precursor to human LIF isoform 1.   
     
     
         14 . The method of  claim 13  wherein the cancer is one of the following: glioma, pre-B cell acute lymphoblastic leukemia, acute myeloid leukemia, lung adenocarcinoma, prostate adenocarcinoma, colorectal carcinoma, bladder cancer, breast ductal cancer or breast carcinoma. 
     
     
         15 . An in vitro method for selecting patients suffering from a cancer, to be treated with a monoclonal antibody or a fragment thereof, which binds full length human LIF, but does not bind a LIF fragment corresponding to amino acids 1 to 160, wherein the full length human LIF is the precursor to human LIF isoform 1, said method comprising
 (a) quantifying the expression levels of LIF in said patient, and   (b) comparing said expression levels with control levels,   wherein if the expression levels of LIF in said patient are greater than the control values, then said patient is selected to receive treatment with said antibody or fragment thereof.   
     
     
         16 . The method of  claim 15 , wherein the cancer is one of the following: glioma, pre-B cell acute lymphoblastic leukemia, acute myeloid leukemia, lung adenocarcinoma, prostate adenocarcinoma, colorectal carcinoma, bladder cancer, breast ductal cancer or breast carcinoma. 
     
     
         17 . An in vitro method for the prognosis of life expectancy or of the probability of survival of subjects suffering from cancer, comprising quantification of the expression levels of LIF in a biological sample from said subject, wherein an increase of LIF expression with respect to LIF expression in a control sample, is indicative of a reduced life expectancy. 
     
     
         18 . The method of  claim 17 , wherein the cancer is one of the following: glioma, pre-B cell acute lymphoblastic leukemia, acute myeloid leukemia, lung adenocarcinoma, prostate adenocarcinoma, colorectal carcinoma, bladder cancer, breast ductal cancer or breast carcinoma. 
     
     
         19 . The method of  claim 17 , wherein quantification of the expression levels of LIF in the biological sample comprises performing a technique selected from Western Blot, immunohistochemistry and ELISA.