MicroRNA Expression in Human Peripheral Blood Microvesicles and Uses Thereof
The present invention provides novel methods and compositions for the diagnosis, prognosis and treatment of disorders by examining samples containing microvesicles and miRs therein.
1 . A method of diagnosing or prognosticating a colorectal cancer in a subject, comprising:
i) determining a level of at least one miR gene product in microvesicles isolated from a sample from the subject, wherein the at least one miR gene product comprises miR-19a, miR-21, miR-127, miR-31, miR-96, miR-135b, miR-183, miR-30c, miR-133a, mir-143, miR-133b and miR-145; and
ii) comparing the level of the at least one miR gene product in the sample to a control, wherein an increase in the level of at least one of miR-19a, miR-21, miR-127, miR-31, miR-96, miR-135b and miR-183, and/or a decrease in the level of at least one of miR-30c, miR-133a, mir-143, miR-133b and miR-145, in the sample from the subject, relative to that of the control, is diagnostic or prognostic of the colorectal cancer.
2 - 33 . (canceled)
34 . The method of claim 1 , wherein the control is selected from the group consisting of:
i) a reference standard;
ii) the level of the at least one miR gene product from a subject that does not have colorectal cancer; and
iii) the level of the at least one miR gene product from a sample of the subject that does not exhibit colorectal cancer.
35 . The method of claim 1 , wherein the subject is a human.
36 - 44 . (canceled)
45 . The method of claim 1 , wherein determining the level of at least one miR gene product comprises:
(a) labeling the miR isolated from the sample from the subject;
(b) hybridizing the miR to an miR array; and
(c) determining miR hybridization to the array.
46 . The method of claim 1 , wherein identifying miR differentially expressed comprises generating an miR profile for the sample and evaluating the miR profile to determine whether miR in the sample are differentially expressed compared to a normal sample.
47 - 50 . (canceled)
51 . The method of claim 1 , wherein the sample comprises a peripheral fluid.
52 . The method of claim 51 , wherein the peripheral fluid comprises blood or a fraction thereof.
53 . The method of claim 1 , wherein determining the level of at least one miR gene product comprises using real-time PCR.
54 . The method of claim 1 , wherein the microvesicles are isolated from the sample using ultracentrifugation and/or flow cytometry.