MICRORNA SIGNATURES INDICATIVE OF IMMUNOMODULATING THERAPY FOR 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, and for monitoring the course of disease or efficacy of treatment, including evaluation of immunomodulating therapy.
1 . A method for evaluating a patient's response to treatment with an immunomodulatory agent used to manage a demyelinating disease, 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 a patient's response to treatment with the immunomodulating agent, the miRNA profile comprising the measured level of at least 4 miRNAs of Table 1, Table 2, and/or Table 3.
2 . The method of claim 1 , wherein the patient has MS, and is being treated with beta-interferon.
3 . The method of claim 1 , wherein the miRNA profile is determined prior to treatment, and after treatment.
4 . The method of claim 1 , wherein the miRNA profile is determined in a serum or plasma sample.
5 - 15 . (canceled)
16 . The method of claim 1 , further comprising, determining the level of one or more normalization controls in the sample.
17 . The method of claim 16 , wherein the sample is spiked with the normalization control(s).
18 . The method of claim 17 , wherein the normalization control is a non-endogenous RNA or miRNA, or a miRNA not detectable in the sample.
19 . (canceled)
20 . (canceled)
21 . The method of claim 1 , wherein miRNA levels are normalized to the Mean or Median detection levels for all miRNAs in the profile.
22 . The method of claim 1 , wherein the miRNA profile is determined by amplification and/or hybridization-based assay.
23 . The method of claim 22 , wherein the miRNA profile is determined by preparing cDNA, followed by Real Time PCR.
24 . (canceled)
25 . The method of claim 1 , wherein the miRNA signature is an algorithm.
26 . The method of claim 25 , wherein the miRNA signature involves threshold miRNA detection levels that are indicative of a patient's response to treatment with beta-interferon.
27 . A method for preparing a miRNA profile indicative of a patient's response to immunomodulating therapy for a demyelinating disease, comprising:
preparing a miRNA profile from a biofluid sample collected from a patient suspected of having MS, the miRNA profile comprising the measured level of 150 miRNAs or less including at least 4 miRNAs of Table 1, Table 2, and/or Table 3.
28 . The method of claim 27 , wherein the patient has MS and is being treated with beta-interferon.
29 . The method of claim 27 , wherein the miRNA profile is determined prior to treatment, and after treatment.
30 . The method of claim 27 , wherein the miRNA profile is determined in a serum or plasma sample.
31 - 42 . (canceled)
43 . The method of claim 27 , further comprising, determining the level of one or more normalization controls in the sample.
44 . The method of claim 43 , wherein the sample is spiked with the normalization control(s).
45 . The method of claim 44 , wherein the normalization control is a non-endogenous RNA or miRNA, or a miRNA not detectable in the sample.
46 . (canceled)
47 . (canceled)
48 . The method of claim 27 , wherein miRNA levels are normalized to the Mean or Median detection level for all miRNAs in the profile.
49 . The method of claim 27 , wherein the miRNA profile is determined by amplification and/or hybridization-based assay.
50 . The method of claim 49 , wherein the miRNA profile is determined by preparing cDNA, followed by Real Time PCR.
51 . (canceled)
52 . The method of claim 27 , wherein the miRNA signature is an algorithm.
53 . The method of claim 52 , wherein the miRNA signature involves threshold miRNA detection levels that are indicative of MS treatment.
54 . A kit for preparing a miRNA profile indicative of a responsive to immunomodulating therapy for a demyelinating disease, comprising: a miRNA-specific primer for reverse transcribing or amplifying each of 150 miRNAs or less, including at least 4 miRNAs of Table 1, Table 2, and/or Table 3.
55 - 71 . (canceled)