MICRORNA PROFILES FOR EVALUATING MULTIPLE SCLEROSIS
The present invention provides methods, systems, and kits for evaluating multiple sclerosis (MS) in a patient. Particularly, the invention provides convenient miRNA-based tests for evaluating a patient for MS, including for diagnosing MS, for excluding MS as a diagnosis, for determining the presence of disease activity associated with MS, and for monitoring the course of disease or efficacy of treatment for MS.
1 . A method for evaluating a patient sample for multiple sclerosis (MS), comprising:
preparing a miRNA profile from a biofluid sample collected from the patient, and determining the presence or absence of a miRNA signature indicative of multiple sclerosis, the miRNA profile comprising the level of at least 4 miRNAs of Table 1, 2, 3, 4, or 5.
2 . The method of claim 1 , wherein the patient is suspected of having MS.
3 . The method of claim 2 , wherein the patient has demyelinating lesions consistent with MS.
4 . The method of claim 2 , wherein the patient has symptoms of a neurologic and/or immunologic disorder consistent with MS.
5 . The method of claim 1 , wherein the miRNA profile is determined prior to treating the patient for MS.
6 . The method of claim 1 , wherein the miRNA profile is indicative of disease activity associated with MS.
7 . The method of claim 1 , wherein the miRNA profile is determined in a serum or plasma sample.
8 . The method of claim 7 , wherein the sample is a serum sample collected with a serum separator tube.
9 . The method of claim 1 , wherein the miRNA profile comprises the level of at least 5 miRNAs of Table 1, 2, or 3.
10 . The method of claim 9 , wherein the miRNA profile comprises the level of at least 6 miRNAs of Table 1, 2, or 3.
11 . The method of claim 9 , wherein the miRNA profile comprises the level of at least 8 miRNAs of Table 1, 2, or 3.
12 . (canceled)
13 . (canceled)
14 . The method of claim 1 , wherein the miRNA profile comprises the level for 150 miRNAs or less.
15 . The method of claim 14 , wherein the miRNA profile comprises the level for 100 miRNAs or less.
16 . (canceled)
17 . (canceled)
18 . (canceled)
19 . (canceled)
20 . The method of claim 1 , wherein the miRNA profile comprises the level for at least one of hsa-miR-125a-3p, hsa-miR-132, hsa-miR-148b, hsa-miR-181a, hsamiR-210, hsa-miR-29c, hsa-miR-31, hsa-miR-331-3p, hsa-miR-335, hsa-miR-375, and hsamiR-483-5p.
21 . (canceled)
22 . The method of claim 20 , wherein the miRNA profile comprises the level for one or more of hsa-miR-29c, hsa-miR-483-5p, hsa-miR-210, hsa-miR-193b, hsa-miR-186, hsamiR-192, hsa-miR-132, and hsa-miR-181a.
23 . (canceled)
24 . The method of claim 1 , further comprising, determining the level of one or more normalization controls in the sample.
25 . The method of claim 24 , wherein the sample is spiked with the normalization control(s).
26 . The method of claim 25 , wherein the normalization control is a non-endogenous RNA or miRNA, or a miRNA not expressed in the sample.
27 . The method of claim 26 , wherein the normalization controls include an Arabidopsis miRNA.
28 . The method of claim 27 , wherein the normalization control(s) include ath-miR-159a.
29 . The method of claim 1 , wherein miRNA levels are normalized to the Mean or Median expression level for all miRNAs in the profile.
30 . The method of claim 1 , wherein miRNA profile is determined by amplication and/or hybridization-based assay.
31 . The method of claim 30 , wherein the miRNA profile is determined by preparing cDNA, followed by Real Time PCR.
32 . The method of claim 31 , wherein the Real Time PCR is TaqMan.
33 . The method of claim 1 , wherein the miRNA signature is an algorithm.
34 . The method of claim 33 , wherein the miRNA signature involves threshold miRNA expression levels that are indicative of MS.
35 . The method of claim 34 , wherein the threshold miRNA levels indicative of MS are set to include the top or bottom 5 to 15% of expression levels within a population of MS patients and healthy controls.
36 . The method of claim 34 , wherein the threshold miRNA levels indicative of MS are above the Mean or Median expression level in samples of a non-MS population.
37 . The method of claim 33 , wherein the miRNA signature involves Mean or Median miRNA expression levels in MS patients as compared to a non-MS population.
38 . The method of claim 1 , wherein the method has a positive predictive value of at least 80%.
39 . The method of claim 38 , wherein the method has a positive predictive value of 90% of greater.
40 . (canceled)
41 . A method for preparing a miRNA profile indicative of the presence or absence of multiple sclerosis (MS), comprising:
preparing a miRNA profile from a biofluid sample collected from a patient suspected of having MS, the miRNA profile comprising the level of 150 miRNAs or less including at least 4 miRNAs of Table 1, 2, 3, 4, or 5.
42 - 69 . (canceled)
70 . A kit for preparing a miRNA profile indicative of the presence or absence of multiple sclerosis (MS), or the presence or absence of disease activity associated with MS, comprising: a miRNA-specific primer for reverse transcribing or amplifying each of 150 miRNAs or less, including at least 4 miRNAs of Table 1, 2, 3, 4, or 5.
71 - 91 . (canceled)