miRNA FINGERPRINT IN THE DIAGNOSIS OF COPD

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 COPD, 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 COPD.   
     
     
         2 . The method according to  claim 1 , wherein the predetermined set of miRNAs comprises miRNAs that are differentially regulated in blood samples from COPD patients as compared to lung cancer patients. 
     
     
         3 . The method according to  claim 1 , wherein the expression profile is determined from miRNAs selected from  FIG. 5  or  FIG. 9 . 
     
     
         4 . The method according to  claim 1 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in  FIG. 10 . 
     
     
         5 . The method according to  claim 1 , wherein the predetermined set of miRNAs comprises miRNAs that are differentially regulated in blood samples from COPD patients as compared to healthy controls. 
     
     
         6 . The method according to  claim 1 , wherein the expression profile is determined from miRNAs selected from  FIG. 3  or  FIG. 7 . 
     
     
         7 . The method according to  claim 1 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in  FIG. 8 . 
     
     
         8 . The method according to  claim 1 , wherein the predetermined set of miRNAs representative for diagnosis of COPD comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 miRNAs. 
     
     
         9 . 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. 
     
     
         10 . A set of polynucleotides for detecting a predetermined set comprising at least two miRNAs for diagnosing and/or prognosing of COPD in a blood sample from a patient, particularly a human patient. 
     
     
         11 . The set of polynucleotides according to  claim 10 , wherein the miRNAs are selected from the miRNAs that are differentially regulated in blood samples from COPD patients as compared to lung cancer patients listed in  FIGS. 5  or  9 . 
     
     
         12 . The set of polynucleotides according to  claim 10 , wherein the predetermined set of miRNAs that are differentially regulated in blood samples from COPD patients as compared to lung cancer patients is selected from the sets of miRNAs listed in  FIG. 10 . 
     
     
         13 . The set of polynucleotides according to  claim 10 , wherein the miRNAs are selected from the miRNAs that are differentially regulated in blood samples from COPD patients as compared to healthy controls listed in  FIG. 3  or  7 . 
     
     
         14 . The set of polynucleotides according to  claim 10 , wherein the predetermined set of miRNAs that are differentially regulated in blood samples from COPD patients as compared to healthy controls is selected from the sets of miRNAs listed in  FIG. 8 . 
     
     
         15 . The set of polynucleotides according to  claim 10 , wherein the predetermined set of miRNAs representative for diagnosis of COPD comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 miRNAs. 
     
     
         16 . Use of set of polynucleotides according to  claim 10  for diagnosing and/or prognosing COPD in a patient, particularly a human patient. 
     
     
         17 . 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 COPD. 
     
     
         18 . The set of primer pairs according to  claim 17 , wherein the miRNAs are selected from the miRNAs that are differentially regulated in blood samples from COPD patients as compared to lung cancer patients listed in  FIG. 5  or  FIG. 9 . 
     
     
         19 . The set of primer pairs according to  claim 17 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in  FIG. 10 . 
     
     
         20 . The set of primer pairs according to  claim 17 , wherein the miRNAs are selected from the miRNAs that are differentially regulated in blood samples from COPD patients as compared to healthy controls listed in  FIG. 3  or  FIG. 7 . 
     
     
         21 . The set of primer pairs according to  claim 17 , wherein the predetermined set of miRNAs that are differentially regulated in blood samples from COPD patients as compared to healthy controls comprises at least one set of miRNAs listed in  FIG. 8 . 
     
     
         22 . The set of primer pairs according to  claim 17 , wherein the predetermined set of miRNAs representative for diagnosis of COPD comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 miRNAs. 
     
     
         23 . Use of set of primer pairs according to  claim 17  for diagnosing and/or prognosing COPD in a patient, particularly a human patient. 
     
     
         24 . Means for diagnosing and/or prognosing of COPD in a blood sample of a subject comprising:
 (i) a set of at least two polynucleotides according to  claim 10  or   (ii) a set of at least two primer pairs which determine the expression levels of a predetermined set of miRNAs.   
     
     
         25 . A kit for diagnosing and/or predicting COPD, 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.   
     
     
         26 . The kit comprising the means according to  claim 24  and at least one reference expression profile for a particular condition. 
     
     
         27 . A set of miRNAs isolated from a blood sample from a subject for diagnosing and/or prognosing of COPD, wherein the miRNAs are selected from the miRNAs as indicated in  claim 2 . 
     
     
         28 . The set of miRNAs of  claim 27  bound to a carrier, e.g. a micro-array. 
     
     
         29 . Use of a set of miRNAs according to  claim 27  for diagnosing and/or prognosing of COPD in a subject, particularly a human subject. 
     
     
         30 . The method of  claim 1 , wherein said sample is a whole blood sample.