Microrna biomarkers indicative of alzheimer's disease
The invention provides a method of diagnosing Alzheimer's Disease in a subject, by determining the level of at least one miRNA in a sample derived from the subject, wherein a change in the level of the at least one miRNA relative to a suitable control is indicative of Alzheimer's Disease in the subject. Methods for monitoring the course of Alzheimer's Disease, methods of treating a subject having Alzheimer's Disease, and kits for diagnosing Alzheimer's Disease are also provided.
1. A method of detecting the level of one or more miRNAs in a plasma or serum sample from a subject known or suspected of having cognitive impairment due to Alzheimer's Disease or Mild Cognitive Impairment (MCI), said method comprising:
providing a plasma or serum sample from the subject known or suspected of having cognitive impairment due to Alzheimer's Disease or MCI; and
detecting the level of at least one miRNA selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, and combinations thereof, in the plasma or serum sample by contacting the plasma or serum sample with an agent that specifically hybridizes to the at least one miRNA, or a cDNA derived therefrom, and detecting the level of hybridization between the agent and the at least one miRNA, or cDNA derived therefrom.
2. The method of claim 1 , comprising detecting the level of one miRNA selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, and Let-7d in the plasma or serum sample.
3. The method of claim 1 , further comprising detecting the level of at least one additional miRNA in the plasma or serum sample selected from the group consisting of miR-301a; miR-545; and miR-301a and miR-545.
4. The method of claim 3 , comprising detecting the level of at least two miRNAs in the sample.
5. The method of claim 3 , comprising detecting the level of miR-545, let-7g, and miR-15b.
6. The method of claim 3 , comprising detecting the level of miR-15b and miR-545.
7. The method of claim 3 , comprising detecting the level of miR-301a, miR-545, let-7g and miR-15b.
8. The method of claim 3 , comprising detecting the level of miR-191, miR-301a, and miR-545.
9. The method of claim 3 , comprising detecting the level of miR-301a, let-7g, and miR-15b.
10. The method of claim 3 , comprising detecting the level of miR-191, miR-301a, miR-545, and miR-15b.
11. The method of claim 3 , comprising detecting the level of at least three miRNAs selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, miR-301a, and miR-545, in the plasma or serum sample.
12. The method of claim 1 , comprising detecting the level of at least two miRNAs in the sample, wherein said miRNAs are selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, and combinations thereof.
13. The method of claim 1 , comprising detecting the level of miR-191 and miR-15b.
14. The method of claim 1 , comprising detecting the level of Let-7g and miR-15b.
15. The method of claim 1 , wherein the plasma or serum sample is a microvesicle-free sample.
16. The method of claim 1 , wherein the sample is plasma.
17. The method of claim 1 , wherein the sample is serum.
18. The method of claim 1 , wherein the subject known or suspected of having cognitive impairment due to Alzheimer's Disease or MCI is selected based on performance in a cognitive test to facilitate diagnosis of Alzheimer's Disease.
19. The method of claim 18 , wherein the cognitive test is selected from the group consisting of the mini-mental state examination (MMSE), mini-cog test, ADAS-cog test, and clock-drawing test.
20. The method of claim 1 , further comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising an Alzheimer's therapeutic or a pharmaceutically acceptable salt or ester thereof.
21. The method of claim 20 , wherein the Alzheimer's therapeutic is selected from the group consisting of galantamine, rivastigmine and donepezil.
22. The method of claim 1 , wherein the agent that specifically hybridizes to the miRNA is a nucleic acid molecule.
23. The method of claim 1 , wherein the level of hybridization between the agent and the at least one miRNA is detected using amplification, hybridization, and/or sequencing methods.
24. The method of claim 23 , wherein the level of hybridization between the agent and the at least one miRNA is detected using polymerase chain reaction (PCR), Real-Time PCR (RT-PCR), quantitative PCR (qPCR), or rolling circle amplification.
25. The method of claim 23 , wherein the level of hybridization between the agent and a the at least one miRNA is detected using microarray, NanoString nCounter analysis, Northern blot, branched DNA signal amplification, or in situ hybridization.
26. The method of claim 23 , wherein the level of hybridization between the agent and the at least one miRNA is detected using next-generation sequencing.
27. The method of claim 1 , wherein the subject is a mammal.
28. The method of claim 1 , wherein the subject is human.
29. A method of detecting the level of one or more miRNAs in a plasma or serum sample from a subject having cognitive impairment with an MMSE score of 0-26, comprising:
providing a plasma or serum sample from the subject having cognitive impairment; and
detecting the level of at least one miRNA selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, and combinations thereof, in the plasma or serum sample by contacting the plasma or serum sample with an agent that specifically hybridizes to the at least one miRNA, or a cDNA derived therefrom, and detecting the level of hybridization between the agent and the at least one miRNA, or cDNA derived therefrom.
30. The method of claim 29 , further comprising detecting the level of at least one additional miRNA in the plasma or serum sample selected from the group consisting of miR-301a; miR-545; and miR-301a and miR-545.
31. A method of detecting the level of one or more miRNAs in a plasma or serum sample from a subject having cognitive impairment with an MMSE score of 21-26, comprising:
providing a plasma or serum sample from the subject having cognitive impairment; and
detecting the level of at least one miRNA selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, and combinations thereof, in the plasma or serum sample by contacting the plasma or serum sample with an agent that specifically hybridizes to the at least one miRNA, or a cDNA derived therefrom, and detecting the level of hybridization between the agent and the at least one miRNA, or cDNA derived therefrom.
32. The method of claim 31 , further comprising detecting the level of at least one additional miRNA in the plasma or serum sample selected from the group consisting of miR-301a; miR-545; and miR-301a and miR-545.
33. A method of detecting the level of one or more miRNAs in a subject known or suspected of having cognitive impairment due to Alzheimer's Disease or Mild Cognitive Impairment (MCI), said method comprising:
providing a plasma or serum sample from the subject known or suspected of having cognitive impairment due to Alzheimer's Disease or MCI; and
detecting the level of at least one miRNA selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, and combinations thereof, in the plasma or serum sample, by contacting the plasma or serum sample with a nucleic acid molecule that specifically hybridizes to the at least one miRNA, or a cDNA derived therefrom, wherein the nucleic acid molecule comprises a sequence complementary to all or a portion of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:7, or SEQ ID NO:8, and detecting the level of hybridization between the nucleic acid molecule and the at least one miRNA, or cDNA derived therefrom.
34. The method of claim 33 , further comprising detecting the level of at least one additional miRNA in the sample selected from the group consisting of miR-301a; miR-545; and miR-301a and miR-545.
35. The method of claim 33 , comprising detecting the level of at least three miRNAs selected from the group consisting of miR-191, miR-15b, miR-142-3p, Let-7g, Let-7d, miR-301a, and miR-545.
36. The method of claim 33 , comprising detecting the level of at least two miRNAs selected from the group consisting of:
(a) miR-545, let7g, and miR-15b,
(b) miR-15b and miR-545,
(c) miR-301a, miR-545, let-7g and miR-15b,
(d) miR-191 and miR-15b,
(e) Let-7g and miR-15b,
(f) miR-191, miR-301a, and miR-545,
(g) miR-301a, let-7g, and miR-15b, and
(h) miR-191, miR-301a, miR-545, and miR-15b.
37. The method of claim 33 , wherein the level of hybridization between the nucleic acid molecule and the at least one miRNA is detected using amplification, hybridization, and/or sequencing methods.