miRNA FINGERPRINT IN THE DIAGNOSIS OF WILMS' TUMOUR

Abstract

MicroRNAs (miRNA) are a recently discovered class of small non-coding RNAs (17-14 nucleotides). Due to their function as regulators of gene expression they play a critical role both in physiological and in pathological processes, such as cancer. The present invention provides novel methods for diagnosing a state of health based on the determination of specific miRNAs that have altered expression levels in different conditions, e.g. disease states compared to healthy controls.

1 . A method of diagnosing Wilms' tumour, comprising the steps
 (a) determining an expression profile of a predetermined set of miRNAs, in a blood sample from a patient, particularly a human patient; and   (b) comparing said expression profile to a reference expression profile,   
       wherein the comparison of said determined expression profile to said reference expression profile allows for the diagnosis of Wilms' tumour. 
     
     
         2 . The method according to  claim 1 , wherein the expression profile is determined from miRNAs selected from  FIG. 3  or  FIG. 4 . 
     
     
         3 . The method according to  claim 1 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in  FIG. 5 . 
     
     
         4 . The method according to  claim 1 , wherein the predetermined set of miRNAs representative for diagnosis of Wilms' tumour comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs. 
     
     
         5 . The method according to  claim 1  wherein the expression profile is determined by nucleic acid hybridization, nucleic acid amplification, polymerase extension, sequencing, mass spectroscopy, flow cytometry or any combinations thereof. 
     
     
         6 . A set of polynucleotides for detecting a predetermined set comprising at least two miRNAs for diagnosing and/or prognosing of Wilms' tumour in a blood sample from a patient, particularly a human patient. 
     
     
         7 . The set of polynucleotides according to  claim 6 , wherein the miRNAs are selected from the miRNAs listed in  FIG. 3  or  4 . 
     
     
         8 . The set of polynucleotides according to  claim 6 , wherein the predetermined set of miRNAs is selected from the sets of miRNAs listed in  FIG. 5 . 
     
     
         9 . The set of polynucleotides according to  claim 6 , wherein the predetermined set of miRNAs representative for diagnosis of Wilms' tumour comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs. 
     
     
         10 . Use of set of polynucleotides according to  claim 6  for diagnosing and/or prognosing Wilms' tumour in a patient, particularly a human patient. 
     
     
         11 . A set of primer pairs for determining the expression level of a predetermined set of miRNAs in a blood sample of a patient for diagnosing and/or prognosing Wilms' tumour. 
     
     
         12 . The set of primer pairs according to  claim 11 , wherein the miRNAs are selected from the miRNAs listed in  FIG. 3  or  FIG. 4 . 
     
     
         13 . The set of primer pairs according to  claim 11 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in  FIG. 5 . 
     
     
         14 . The set of primer pairs according to  claim 11 , wherein the predetermined set of miRNAs representative for diagnosis of Wilms' tumour comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs. 
     
     
         15 . Use of set of primer pairs according to  claim 11  for diagnosing and/or prognosing Wilms' tumour in a patient, particularly a human patient. 
     
     
         16 . Means for diagnosing and/or prognosing of Wilms' tumour in a blood sample of a subject comprising:
 a set of at least two polynucleotides according to  claim 6  or   a set of at least two primer pairs which comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs.   
     
     
         17 . A kit for diagnosing and/or predicting Wilms' tumour, comprising:
 (a) means for determining the miRNA expression profile of a RNA sample of a subject, and   (b) at least one reference expression profile for a particular condition.   
     
     
         18 . (canceled) 
     
     
         19 . A set of miRNAs isolated from a blood sample from a subject for diagnosing and/or prognosing of Wilms' tumour, wherein the miRNAs are selected from the miRNAs as indicated in  claim 2 . 
     
     
         20 . The set of miRNAs of  claim 19  bound to a carrier, e.g. a micro-array. 
     
     
         21 . Use of a set of miRNAs according to  claim 19  for diagnosing and/or prognosing of Wilms' tumour in a subject, particularly a human subject. 
     
     
         22 . The method of  claim 1 , wherein said sample is a whole blood sample.