MIRNAS AS BIOMARKERS FOR DISTINGUISHING BENIGN FROM MALIGNANT THYROID NEOPLASMS
The present invention concerns methods and compositions for identifying a miRNA profile for a particular condition, such as thyroid nodules or thyroid cancer, and using the profile in the diagnosis of a patient for a condition, such as thyroid nodules or thyroid cancer.
1 - 34 . (canceled)
35 . A method of treating a subject identified with a thyroid carcinoma, the method comprising:
producing labeled nucleotide sequences from a thyroid sample from a subject;
contacting the thyroid sample from the subject with labeled primers specific for a group of microRNAs comprising miR-21, miR-31, miR-146b-5p, miR-222, miR-375, miR-551b, miR-29b-2, miR-138, miR-139-5p, miR-155, and miR-204; and
reacting the sample with the labeled primers under conditions to amplify the microRNAs using polymerase chain reaction to produce the labeled nucleotide sequences;
measuring expression levels of microRNAs miR-21, miR-31, miR-146b-5p, miR-222, miR-375, miR-551b, miR-29b-2, miR-138, miR-139-5p, miR-155, and miR-204 from the unclassified thyroid sample from the subject with the labeled modified nucleotide microRNA probes that bind specifically to miR-21, miR-31, miR-146b-5p, miR-222, miR-375, miR-551b, miR-29b-2, miR-138, miR-139-5p, miR-155, and miR-204 to produce a microRNA profile based upon the binding of the labeled nucleotide microRNA probes and the labeled nucleotide sequences produced using the polymerase chain reaction;
comparing the expression levels of the microRNAs in the profile from the unclassified thyroid sample from the subject to a normal thyroid reference level for each miRNA;
identifying the unclassified thyroid sample as thyroid carcinoma when the profile expression levels of miR-21, miR-31, miR-146b-5p, miR-222, miR-375, miR-29b-2, miR-138, miR-139-5p, and miR-155 are increased as compared to the normal reference level and miR-551b, and miR-204 are decreased as compared to the normal reference level; and
treating the subject whose sample was identified as thyroid carcinoma.
36 . The method of claim 35 , wherein the group of microRNAs further comprises miR-885-5p.
37 . The method of claim 35 , wherein the sample is a biopsy.
38 . The method of claim 37 , wherein the biopsy is surgical resection or a fine needle aspirate.
39 . The method of claim 35 , further comprising obtaining a sample from the subject.
40 . The method of claim 35 , wherein the treating comprises administering radioactive iodine, radiation, or chemotherapy to the subject identified as having papillary thyroid carcinoma or performing a thyroidectomy on the subject identified as having papillary thyroid carcinoma.
41 . A method of producing labeled nucleotide sequences from a thyroid sample from a subject, the method comprising
contacting the thyroid sample from the subject with labeled primers specific for a group of microRNAs comprising miR-21, miR-31, miR-146b-5p, miR-222, miR-375, miR-551b, miR-29b-2, miR-138, miR-139-5p, miR-155, and miR-204; and
reacting the sample with the labeled primers under conditions to amplify the microRNAs using polymerase chain reaction to produce the labeled nucleotide sequences.
42 . The method of claim 41 , wherein the group of microRNAs further comprises miR-885-5p.
43 . The method of claim 41 , wherein the label is non-radioactive.
44 . The method of claim 41 , wherein the label is radioactive.
45 . The method of claim 41 , wherein the label is a UV spectrum emitting label.
46 . The method of claim 41 , wherein the label is a fluorescent label.
47 . The method of claim 41 , wherein the label is an enzymatic label.
48 . A composition comprising labeled thyroid derived nucleic acid molecules, wherein the labeled molecules comprise nucleic acid molecules that hybridize with or correspond to miR-21, miR-31, miR-146b-5p, miR-222, miR-375, miR-551b, miR-29b-2, miR-138, miR-139-5p, miR-155, and miR-204.
49 . The composition of claim 48 , wherein the labeled molecules further comprise nucleic acid molecules that hybridize with or correspond to miR-885-5p.
50 . The composition of claim 48 , wherein the label is non-radioactive.
51 . The composition of claim 48 , wherein the label is radioactive.
52 . The composition of claim 48 , wherein the label is a UV spectrum emitting label.
53 . The composition of claim 48 , wherein the label is a fluorescent label.
54 . The composition of claim 48 , wherein the label is an enzymatic label.