Complex sets of miRNAs as non-invasive biomarkers for dilated cardiomyopathy
The present invention relates to non-invasive methods, kits and means for diagnosing and/or prognosing of dilated cardiomyopathy in a body fluid sample from a subject. Further, the present invention relates to set of polynucleotides or sets of primer pairs for detecting sets of miRNAs for diagnosing and/or prognosing of dilated cardiomyopathy in a body fluid sample from a subject. In addition, the present invention relates to sets of miRNAs for diagnosing and/or prognosing of dilated cardiomyopathy in a body fluid sample from a subject.
1. A method of detecting an expression profile for dilated cardiomyopathy in a subject comprising the steps of:
(i) providing a blood cell sample comprising a mixture of erythrocytes, leukocytes, and thrombocytes obtained from the subject;
(ii) detecting whether the expression profile for dilated cardiomyopathy is present in the blood cell sample obtained from the subject by contacting the blood cell sample with probes for a miRNA expression profile and detecting binding between the probes and the miRNAs,
wherein the expression profile comprises a set of at least two miRNAs selected from the group consisting of:
1) miR-520d-5p having SEQ ID No: 1, and
2) at least one additional miRNA selected from the group consisting of:
miR-558 having SEQ ID NO: 6,
miR-122* having SEQ ID NO: 3,
miR-622 having SEQ ID NO: 24,
miR-519e* having SEQ ID NO: 34,
miR-1228* having SEQ ID NO: 30,
miR-200b* having SEQ ID NO: 14, and
miR-1231 having SEQ ID NO: 27; and
(iii) identifying whether the subject has dilated cardiomyopathy by transforming the individual expression levels of the miRNAs in the individual blood cells of the blood cell sample to one numerical value which represents the mathematical average of the miRNA expression levels of the individual blood cells and comparing the expression profile to a reference expression profile obtained from at least one healthy human subject or a human subject not having dilated cardiomyopathy, whereby the difference between expression profile and the reference expression profile is indicative of dilated cardiomyopathy.
2. The method of claim 1 , wherein the set of miRNAs comprises at least one of the sets of miRNAs selected from the group consisting of:
DCM-39: hsa-miR-558, hsa-miR-122*, hsa-miR -520d-5p, hsa-miR -622, hsa-miR-519e*, hsa-miR -1228*, hsa-miR -200b*, hsa-miR -1231;
DCM-40: hsa-miR-122*, hsa-miR-1228*;
DCM-41: hsa-miR-122*, hsa-miR-558;
DCM-42: hsa-miR-122*, hsa-miR-519e;
DCM-43: hsa-miR-122*; hsa-miR-622;
DCM-44: hsa-miR-122*, hsa-miR-520d-5p;
DCM-45: hsa-miR-122*, hsa-miR-200b*;
DCM-46: hsa-miR-1228*, hsa-miR-558;
DCM-47: hsa-miR-1228*, hsa-miR-519e*;
DCM-48: hsa-miR-1228*, hsa-miR-622;
DCM-49: hsa-miR-1228*, hsa-miR-520d-5p;
DCM-50: hsa-miR-1228*, hsa-miR-200b*;
DCM-51: hsa-miR-558, hsa-miR-519e*;
DCM-52: hsa-miR-558, hsa-miR-622;
DCM-53: hsa-miR-558, hsa-miR-520d-5p;
DCM-54: hsa-miR-558, hsa-miR-200b*;
DCM-55: hsa-miR-519e*, hsa-miR-622;
DCM-56: hsa-miR-519e*, hsa-miR-520d-5p;
DCM-57: hsa-miR-519e*, hsa-miR-200b*;
DCM-58: hsa-miR-622, hsa-miR-520d-5p;
DCM-59: hsa-miR-622, hsa-miR-200b*;
DCM-60: hsa-miR-122*, hsa-miR-1228*, hsa-miR-558;
DCM-61: hsa-miR-122*, hsa-miR-1228*, hsa-miR-519e*;
DCM-62: hsa-miR-122*, hsa-miR-1228*, hsa-miR-622;
DCM-63: hsa-miR-122*, hsa-miR-1228*, hsa-miR-520d-5p;
DCM-64: hsa-miR-122*, hsa-miR-1228*, hsa-miR-200b*;
DCM-65: hsa-miR-122*, hsa-miR-558, hsa-miR-519e*;
DCM-66: hsa-miR-122*, hsa-miR-558, hsa-miR-622;
DCM-67: hsa-miR-122*, hsa-miR-558, hsa-miR-520d-5p;
DCM-68: hsa-miR-122*, hsa-miR-558, hsa-miR-200b*;
DCM-69: hsa-miR-122*, hsa-miR-519e*, hsa-miR-622;
DCM-70: hsa-miR-122*, hsa-miR-519e*, hsa-miR-520d-5p;
DCM-71: hsa-miR-122*, hsa-miR-519e*, hsa-miR-200b*;
DCM-72: hsa-miR-122*, hsa-miR-622, hsa-miR-520d-5p;
DCM-73: hsa-miR-122*, hsa-miR-622, hsa-miR-200b*;
DCM-74: hsa-miR-122*, hsa-miR-520d-5p, hsa-miR-200b*;
DCM-75: hsa-miR-1228*, hsa-miR-558, hsa-miR-519e*;
DCM-76: hsa-miR-1228*, hsa-miR-558, hsa-miR-622;
DCM-77: hsa-miR-1228*, hsa-miR-558, hsa-miR-520d-5p;
DCM-78: hsa-miR-1228*, hsa-miR-558, hsa-miR-200b*;
DCM-79: hsa-miR-1228*, hsa-miR-519e*, hsa-miR-622;
DCM-80: hsa-miR-1228*, hsa-miR-519e*, hsa-miR-520d-5p;
DCM-81: hsa-miR-1228*, hsa-miR-519e*, hsa-miR-200b*;
DCM-82: hsa-miR-1228*, hsa-miR-622, hsa-miR-520d-5p;
DCM-83: hsa-miR-1228*, hsa-miR-622, hsa-miR-200b*;
DCM-84: hsa-miR-1228*, hsa-miR-520d-5p, hsa-miR-200b*;
DCM-85: hsa-miR-558, hsa-miR-519e*, hsa-miR-622;
DCM-86: hsa-miR-558, hsa-miR-519e*, hsa-miR-520d-5p;
DCM-87: hsa-miR-558, hsa-miR-519e*, hsa-miR-200b*;
DCM-88: hsa-miR-558, hsa-miR-622, hsa-miR-520d-5p;
DCM-89: hsa-miR-558, hsa-miR-622, hsa-miR-200b*;
DCM-90: hsa-miR-558, hsa-miR-520d-5p, hsa-miR-200b*;
DCM-91: hsa-miR-519e*, hsa-miR-622, hsa-miR-520d-5p;
DCM-92: hsa-miR-519e*, hsa-miR-622, hsa-miR-200b*;
DCM-93: hsa-miR-519e*, hsa-miR-520d-5p, hsa-miR-200b*; and
DCM-94: hsa-miR-622, hsa-miR-520d-5p, hsa-miR-200b*.