Method for detecting nucleosomes containing histone variants

Abstract

The invention relates to a method for detecting and measuring the presence of mono-nucleosomes and oligo-nucleosomes and nucleosomes that contain particular histone variants and the use of such measurements for the detection and diagnosis of disease. The invention also relates to a method of identifying histone variant biomarkers for the detection and diagnosis of disease and to biomarkers identified by said method.

1 . A method of treating a disease that is colitis, chronic obstructive pulmonary disorder or Crohn's disease, comprising:
 (a) obtaining a body fluid sample from a patient;   (b) detecting a level of cell free nucleosomes comprising a histone variant or histone isoform in the body fluid sample comprising contacting the body fluid sample with a binding agent which binds to the histone variant or histone isoform;   (c) identifying the patient as suffering from the disease based on the level of cell free nucleosomes comprising a histone variant or histone isoform detected; and   (d) administering a treatment to the patient identified as suffering from the disease.   
     
     
         2 . The method as defined in  claim 1 , wherein the cell free nucleosome is a mononucleosome or an oligonucleosome. 
     
     
         3 . The method as defined in  claim 1 , wherein the body fluid sample is a blood, serum or plasma sample. 
     
     
         4 . The method as defined in  claim 1 , wherein the histone variant or histone isoform is an isoform of histone H2A, H2B or H3. 
     
     
         5 . The method as defined in  claim 4 , wherein the binding agent is an antibody. 
     
     
         6 . The method as defined in  claim 1 , wherein the step of detecting or measuring comprises:
 (i) contacting the body fluid sample with a first binding agent which binds to (1) nucleosomes or (2) the histone variant or histone isoform;   (ii) contacting the sample bound in step (i) with a second binding agent which binds to (1) the histone variant or histone isoform when the first binding agent binds to nucleosomes, or (2) nucleosomes when the first binding agent binds to the histone variant or histone isoform; and   (iii) detecting or quantifying the binding of said second binding agent in the body fluid sample.   
     
     
         7 . The method as defined in  claim 6 , wherein the first binding agent and/or the second binding agent is an antibody. 
     
     
         8 . The method as defined in  claim 1 , wherein the step of detecting or measuring comprises:
 (i) contacting the body fluid sample with a first binding agent which binds to nucleosomes;   (ii) contacting the sample bound in step (i) with a second binding agent which binds to the histone variant or histone isoform; and   (iii) detecting or quantifying the binding of said second binding agent to histone variant or histone isoform in the body fluid sample.   
     
     
         9 . The method as defined in  claim 8 , wherein the first binding agent and/or the second binding agent is an antibody. 
     
     
         10 . The method as defined in  claim 1 , wherein the step of detecting or measuring comprises:
 (i) contacting the body fluid sample with a first binding agent which binds to the histone variant or histone isoform;   (ii) contacting the sample bound in step (i) with a second binding agent which binds to nucleosomes; and   (iii) detecting or quantifying the binding of said second binding agent to nucleosomes in the body fluid sample.   
     
     
         11 . The method as defined in  claim 10 , wherein the first binding agent and/or the second binding agent is an antibody. 
     
     
         12 . The method as defined in  claim 1 , wherein the histone variant or histone isoform associated with a cell free nucleosome is detected or measured as one of a panel of measurements. 
     
     
         13 . The method as defined in  claim 12 , wherein the panel comprises a combination of one or more histone variants or isoforms, and/or one or more DNA bases, and/or one or more histone modifications.