Mirna fingerprint in the diagnosis of lung cancer

Abstract

The present invention provides novel methods for diagnosing diseases based on the determination of specific miRNAs that have altered expression levels in disease states compared to healthy controls.

1 .- 24 . (canceled) 
     
     
         25 . A method for diagnosing lung cancer in a patient comprising the steps of:
 (a) determining an expression profile of a predetermined set of miRNAs in a blood sample from a patient, wherein the predetermined set of miRNAs comprises one or more miRNAs selected from the group consisting of hsa-miR-15a, hsa-miR-126, hsa-miR-195, hsa-miR-26b, hsa-miR-19a, and hsa-miR-18a*, 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 lung cancer.   
     
     
         26 . The method of  claim 25 , wherein said predetermined set of miRNAs comprises or consists of.
 (i) hsa-miR-22, hsa-miR-15a, and hsa-miR-98,   (ii) hsa-let-7i, hsa-let-7d, hsa-miR-22, hsa-miR-15a, and hsa-miR-98,   (iii) hsa-miR-22, hsa-miR-15a, hsa-miR-98, hsa-miR-19a, and hsa-miR-574-5p,   (iv) hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, and hsa-miR-324-3p,   (v) hsa-miR-126, hsa-miR-423-5p, hsa-let-7i, hsa-let-7d, hsa-miR-22, hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, and hsa-miR-324-3p,   (vi) hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, hsa-miR-324-3p, hsa-miR-20b, hsa-miR-25, hsa-miR-195, hsa-let-7e, hsa-let-7c, and hsa-let-7f,   (vii) hsa-let-7d, hsa-miR-15a, hsa-miR-19a, hsa-miR-324-3p, and hsa-miR-25,   (viii) hsa-miR-1909, hsa-let-7c, and hsa-miR-15a,   (ix) hsa-miR-15a, hsa-miR-93*, and hsa-miR-30e,   (x) hsa-miR-299-3p, hsa-miR-423-3p, hsa-miR-18a*, hsa-miR-1909, hsa-let-7c, and hsa-miR-15a,   (xi) hsa-miR-1909, hsa-let-7c, hsa-miR-15a, hsa-miR-425, hsa-miR-93*, and hsa-miR-665,   (xii) hsa-miR-604, hsa-miR-22, hsa-let-7b, hsa-miR-299-3p, hsa-miR-423-3p, hsa-miR-18a*, hsa-miR-1909, hsa-let-7c, hsa-miR-15a, and hsa-miR-425, or   (xiii) hsa-miR-18a*, hsa-miR-1909, hsa-let-7c, hsa-miR-15a, hsa-miR-425, hsa-miR-93*, hsa-miR-665, hsa-miR-30e, hsa-miR-339-3p, and hsa-miR-1307.   
     
     
         27 . The method of  claim 25 , wherein the blood sample is blood cell sample. 
     
     
         28 . The method of  claim 27 , wherein the blood cell sample comprises erythrocytes, leukocytes and thrombocytes. 
     
     
         29 . The method of  claim 25 , wherein the expression profile of the predetermined set of miRNAs is determined using reverse-transcribed miRNAs. 
     
     
         30 . The method of  claim 25 , wherein the determination of the expression profile of the predetermined set of miRNAs comprises the steps of:
 (i) extracting total RNA from said blood sample,   (ii) reverse-transcribing the total RNA into cDNA, and   (iii) amplifying the cDNA and thereby quantifying said miRNAs.   
     
     
         31 . The method of  claim 25 , wherein the diagnosis comprises determining type, grade, and/or stage of cancer. 
     
     
         32 . The method of  claim 25 , wherein the diagnosis comprises determining survival rate, responsiveness to drugs, and/or monitoring the course of the disease or the therapy, e.g. chemotherapy, staging of the disease, measuring the response of a patient to therapeutic intervention, segmentation of patients suffering from the disease, identifying of a patient who has a risk to develop the disease, predicting/estimating the occurrence, preferably the severity of the occurrence of the disease, predicting the response of a patient with the disease to therapeutic intervention. 
     
     
         33 . The method of  claim 25 , wherein the lung cancer selected from the group consisting of lung carcinoid, lung pleural mesothelioma and lung squamous cell carcinoma, in particular non-small cell lung carcinoma. 
     
     
         34 . The method of  claim 25 , wherein the determination of an expression profile in step (a) comprises nucleic acid hybridization, nucleic acid amplification, polymerase extension, sequencing, mass spectroscopy or any combinations thereof, wherein the nucleic acid hybridization is particularly performed using a solid-phase nucleic acid biochip array, in particular a microarray, a bead-based assay, or in situ hybridization or wherein the nucleic acid amplification method is real-time PCR (RT-PCR). 
     
     
         35 . A kit for diagnosing lung cancer comprising:
 (a) means for determining an expression profile of a predetermined set of miRNAs in a blood sample from a patient, wherein the predetermined set of miRNAs comprises one or more miRNAs selected from the group consisting of hsa-miR-15a, hsa-miR-126, hsa-miR-195, hsa-miR-26b, hsa-miR-19a, and hsa-miR-18a*, and   (b) a reference expression profile of said predetermined set of miRNAs in a blood sample from a healthy subject.   
     
     
         36 . The kit of  claim 35 , wherein said predetermined set of miRNAs comprises, essentially consists of or consists of.
 (i) hsa-miR-22, hsa-miR-15a, and hsa-miR-98,   (ii) hsa-let-7i, hsa-let-7d, hsa-miR-22, hsa-miR-15a, and hsa-miR-98,   (iii) hsa-miR-22, hsa-miR-15a, hsa-miR-98, hsa-miR-19a, and hsa-miR-574-5p,   (iv) hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, and hsa-miR-324-3p,   (v) hsa-miR-126, hsa-miR-423-5p, hsa-let-7i, hsa-let-7d, hsa-miR-22, hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, and hsa-miR-324-3p,   (vi) hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, hsa-miR-324-3p, hsa-miR-20b, hsa-miR-25, hsa-miR-195, hsa-let-7e, hsa-let-7c, and hsa-let-7f,   (vii) hsa-let-7d, hsa-miR-15a, hsa-miR-19a, hsa-miR-324-3p, and hsa-miR-25,   (viii) hsa-miR-1909, hsa-let-7c, and hsa-miR-15a,   (ix) hsa-miR-15a, hsa-miR-93*, and hsa-miR-30e,   (x) hsa-miR-299-3p, hsa-miR-423-3p, hsa-miR-18a*, hsa-miR-1909, hsa-let-7c, and hsa-miR-15a,   (xi) hsa-miR-1909, hsa-let-7c, hsa-miR-15a, hsa-miR-425, hsa-miR-93*, and hsa-miR-665,   (xii) hsa-miR-604, hsa-miR-22, hsa-let-7b, hsa-miR-299-3p, hsa-miR-423-3p, hsa-miR-18a*, hsa-miR-1909, hsa-let-7c, hsa-miR-15a, and hsa-miR-425, or   (xiii) hsa-miR-18a*, hsa-miR-1909, hsa-let-7c, hsa-miR-15a, hsa-miR-425, hsa-miR-93*, hsa-miR-665, hsa-miR-30e, hsa-miR-339-3p, and hsa-miR-1307.