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) isolating microvesicles from a sample from the subject;
ii) determining a level of at least one miR gene product in the microvesicles, 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 or miR-145; and
iii) comparing the level of the at least one miR gene product in the sample to a control;
iv) detecting an increased level of the at least one miR gene product in the sample compared to the control, wherein the at least one miR gene product comprises miR-19a, miR-21, miR-127, miR-31, miR-96, miR-135b or miR-183, and/or
detecting a decreased level of the at least one miR gene product in the sample compared to the control, wherein the at least one miR gene product comprises miR-30c, miR-133a, miR-143, miR-133b, or miR-145;and
v) diagnosing or prognosticating colorectal cancer from the detected increased level of the at least one miR gene product that comprises miR-19a, miR-21, miR-127, miR-31, miR-96, miR-135b or miR-183, or from the decreased level of the at least one miR gene product that comprises miR-30c, miR-133a, miR-143, miR-133b, or miR-145.
2. 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.
3. The method of claim 1 , wherein the subject is a human.
4. 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.
5. The method of claim 1 , wherein the comparing and detecting comprise 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.
6. The method of claim 1 , wherein the sample comprises a peripheral fluid.
7. The method of claim 6 , wherein the peripheral fluid comprises blood or a fraction thereof.
8. The method of claim 1 , wherein determining the level of at least one miR gene product comprises using real-time PCR.
9. The method of claim 1 , wherein the microvesicles are isolated from the sample using ultracentrifugation and/or flow cytometry.
10. The method of claim 1 , wherein the determining step comprises determining a level of 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.
11. The method according to claim 1 , further comprising administering at least one therapeutic agent to the subject to modulate the level of the at least one miR gene product in the subject.
12. The method according to claim 11 , wherein the at least one miR gene product comprises miR-30c, miR-133a, miR-143, miR-133b, or miR-145,
wherein a decreased level of the at least one miR gene product has been detected in the sample, and
wherein the administering step comprises administering, as the therapeutic agent, an effective amount of said miR gene product to the subject.
13. The method according to claim 11 , wherein the at least one miR gene product comprises miR-19a, miR-21, miR-127, miR-31, miR-96, miR-135b or miR-183,
wherein an increased level of the at least one miR gene product has been detected in the sample, and
wherein the administering step comprises administering to the subject, as the therapeutic agent, an effective amount of a compound that inhibits expression of said miR gene product in the subject, said compound comprising at least one member selected from the group consisting of double-stranded RNA, antisense nucleic acid, and enzymatic RNA.
14. A method of diagnosing or prognosticating a colorectal cancer in a subject, comprising:
i) isolating microvesicles from a sample from the subject;
ii) determining a level of at least one miR gene product in the microvesicles, wherein the at least one miR gene product comprises miR-127, miR-30c, miR-133a, or miR-143; and
iii) comparing the level of the at least one miR gene product in the sample to a control;
iv) detecting an increased level of the at least one miR gene product in the sample compared to the control, wherein the at least one miR gene product comprises miR-127and/or
detecting a decreased level of the at least one miR gene product in the sample compared to the control, wherein the at least one miR gene product comprises miR-30c, miR-133a, or miR-143; and
v) diagnosing or prognosticating colorectal cancer from the detected increased level of the at least one miR gene product that comprises miR-127, or from the decreased level of the at least one miR gene product that comprises miR-30c, miR-133a, or miR-143.