miRNA BIOMARKERS OF LUNG DISEASE

Abstract

This application describes miRNAs that may be used as serum or plasma biomarkers for characterizing lung disease in a patient. These miRNA biomarkers may be used alone or in combination with other markers for the diagnosis, prognosis, or monitoring of diseases such as lung cancer.

1 - 19 . (canceled) 
     
     
         20 . A method of characterizing a lung disease in a patient as cancerous or benign, comprising the steps of:
 a. reverse transcribing a miRNA in a serum sample;   b. amplifying the miRNA;   c. measuring the level of the miRNA;   d. detecting whether the level of the miRNA is elevated or reduced,   thereby characterizing the lung disease as cancerous if the miRNA is elevated or reduced,   wherein the miRNA is chosen from miR-210, miR-155, miR-142-5p, miR-181d, miR-27b, miR-15b, miR-126, miR-21, miR-205, let-7b, let-7c, let-7d, let-7e, let-7f, let-7g, let-7i, miR-10a, miR-100b, miR-130b, miR-132, miR-133b, miR-139, miR-143, miR-152, miR-17-5p, miR-193, miR-194, miR-195, miR-196b, miR-199a*, miR-19b, miR-202, miR-204, miR-206, miR-20b, miR-214, miR-221, miR-27a, miR-296, miR-29a, miR-301, miR-324-3p, miR-324-5p, miR-339, miR-346, miR-365, miR-378, miR-422a, miR-432, miR-485-3p, miR-496, miR-497, miR-505, miR-518b, miR-525, miR-566, miR-605, miR-638, miR-660, miR-93, miR-101, miR-106a, miR-106b, miR-125a, miR-140, miR-145, miR-146a, miR-146b, miR-150, miR-151, miR-15a, miR-181a, miR-181b, miR-185, miR-186, miR-190, miR-191, miR-222, miR-23a, miR-24, miR-26a, miR-26b, miR-30a-5p, miR-30b, miR-30c, miR-30d, miR-30e-5p, miR-320, miR-335, miR-340, miR-342, miR-345, and miR-375.   
     
     
         21 . The method of  claim 20 , wherein the lung cancer is small cell or non-small cell lung cancer. 
     
     
         22 . The method of  claim 20 , wherein the amplification is by quantitative reverse transcriptase polymerase chain reaction. 
     
     
         23 . The method of  claim 20 , wherein the reverse transcription comprises a reverse transcription primer that includes a portion complementary to the miRNA and a 5′ non-complementary portion. 
     
     
         24 . The method of  claim 20 , wherein the amplification comprises a first primer that includes a portion complementary to a cDNA copy of the miRNA and a 5′ non-complementary portion. 
     
     
         25 . The method of  claim 24 , wherein the amplification comprises a second primer complementary to the 5′ non-complementary portion of the first primer. 
     
     
         26 . The method of  claim 20 , further comprising reverse transcribing, amplifying, measuring, and detecting a reduced or elevated level of a second miRNA in the serum sample. 
     
     
         27 . The method of  claim 20 , wherein the patient has a lung tumor or lesion. 
     
     
         28 . A method of characterizing a lung disease in a patient, comprising the steps of:
 a. reverse transcribing a first miRNA and a second miRNA in a serum sample;   b. amplifying the first miRNA and the second miRNA;   c. measuring the levels of the first miRNA and the second miRNA;   d. detecting whether the levels of the first miRNA and the second miRNA are elevated or reduced,   thereby characterizing the lung disease in the patient,   wherein the first and second miRNA are chosen from miR-210, miR-155, miR-142-5p, miR-181d, miR-27b, miR-15b, miR-126, miR-21, miR-205, let-7a, let-7b, let-7d, let-7f, let-7g, let-7i, miR-101, miR-106a, miR-106b, miR-125a, miR-126*, miR-130b, miR-132, miR-133b, miR-140, miR-145, miR-146a, miR-146b, miR-148b, miR-150, miR-151, miR-15a, miR-181a, miR-181b, miR-185, miR-186, miR-190, miR-191, miR-193a, miR-199a*, miR-202, miR-214, miR-222, miR-23a, miR-24, miR-26a, miR-26b, miR-27a, miR-29b, miR-301, miR-30a-5p, miR-30b, miR-30c, miR-30d, miR-30e-5p, miR-320, miR-324-3p, miR-326, miR-335, miR-340, miR-342, miR-345, miR-346, miR-34a, miR-374, miR-375, miR-378, miR-422a, miR-422b, miR-425, miR-486, miR-496, miR-518b, miR-660, miR-7, miR-92, miR-93, miR-98, miR-99a, and miR-99b.   
     
     
         29 . The method of  claim 28 , wherein the patient has a lung tumor or lesion and the method comprises characterizing the tumor or lesion as cancerous or benign based on the levels of the first and second miRNAs. 
     
     
         30 . The method of  claim 28 , wherein a prognosis or aggressiveness of the lung disease is characterized based on the levels of the first and second miRNAs. 
     
     
         31 . The method of  claim 28 , wherein a progression of the lung disease is characterized based on the levels of the first and second miRNAs. 
     
     
         32 . The method of  claim 28 , wherein the amplification is by quantitative reverse transcriptase polymerase chain reaction. 
     
     
         33 . The method of  claim 28 , wherein the reverse transcription comprises a reverse transcription primer that includes a portion complementary to the first or second miRNA and a 5′ non-complementary portion. 
     
     
         34 . The method of  claim 28 , wherein the amplification comprises a first primer that includes a portion complementary to a cDNA copy of the first or second miRNA and a 5′ non-complementary portion. 
     
     
         35 . The method of  claim 34 , wherein the amplification comprises a second primer complementary to the 5′ non-complementary portion of the first primer. 
     
     
         36 . The method of  claim 28 , wherein the first and second miRNA are expressed at elevated levels in a patient with lung cancer. 
     
     
         37 . The method of  claim 28 , wherein the first and second miRNA are expressed at reduced levels in a patient with lung cancer. 
     
     
         38 . The method of  claim 28 , wherein the first miRNA is expressed at an elevated level and the second miRNA is expressed at a reduced level in a patient with lung cancer.