miRNA FINGERPRINT IN THE DIAGNOSIS OF WILMS' TUMOUR
MicroRNAs (miRNA) are a recently discovered class of small non-coding RNAs (17-14 nucleotides). Due to their function as regulators of gene expression they play a critical role both in physiological and in pathological processes, such as cancer. The present invention provides novel methods for diagnosing a state of health based on the determination of specific miRNAs that have altered expression levels in different conditions, e.g. disease states compared to healthy controls.
1 . A method of diagnosing Wilms' tumour, comprising the steps
(a) determining an expression profile of a predetermined set of miRNAs, in a blood sample from a patient, particularly a human patient; and
(b) comparing said expression profile to a reference expression profile,
wherein the comparison of said determined expression profile to said reference expression profile allows for the diagnosis of Wilms' tumour.
2 . The method according to claim 1 , wherein the expression profile is determined from miRNAs selected from FIG. 3 or FIG. 4 .
3 . The method according to claim 1 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in FIG. 5 .
4 . The method according to claim 1 , wherein the predetermined set of miRNAs representative for diagnosis of Wilms' tumour comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs.
5 . The method according to claim 1 wherein the expression profile is determined by nucleic acid hybridization, nucleic acid amplification, polymerase extension, sequencing, mass spectroscopy, flow cytometry or any combinations thereof.
6 . A set of polynucleotides for detecting a predetermined set comprising at least two miRNAs for diagnosing and/or prognosing of Wilms' tumour in a blood sample from a patient, particularly a human patient.
7 . The set of polynucleotides according to claim 6 , wherein the miRNAs are selected from the miRNAs listed in FIG. 3 or 4 .
8 . The set of polynucleotides according to claim 6 , wherein the predetermined set of miRNAs is selected from the sets of miRNAs listed in FIG. 5 .
9 . The set of polynucleotides according to claim 6 , wherein the predetermined set of miRNAs representative for diagnosis of Wilms' tumour comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs.
10 . Use of set of polynucleotides according to claim 6 for diagnosing and/or prognosing Wilms' tumour in a patient, particularly a human patient.
11 . A set of primer pairs for determining the expression level of a predetermined set of miRNAs in a blood sample of a patient for diagnosing and/or prognosing Wilms' tumour.
12 . The set of primer pairs according to claim 11 , wherein the miRNAs are selected from the miRNAs listed in FIG. 3 or FIG. 4 .
13 . The set of primer pairs according to claim 11 , wherein the predetermined set of miRNAs comprises at least one set of miRNAs listed in FIG. 5 .
14 . The set of primer pairs according to claim 11 , wherein the predetermined set of miRNAs representative for diagnosis of Wilms' tumour comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs.
15 . Use of set of primer pairs according to claim 11 for diagnosing and/or prognosing Wilms' tumour in a patient, particularly a human patient.
16 . Means for diagnosing and/or prognosing of Wilms' tumour in a blood sample of a subject comprising:
a set of at least two polynucleotides according to claim 6 or
a set of at least two primer pairs which comprises at least 1, 7, 10, 15, 20, 25, 30, 35, 40, 50, 75, 100 of miRNAs.
17 . A kit for diagnosing and/or predicting Wilms' tumour, comprising:
(a) means for determining the miRNA expression profile of a RNA sample of a subject, and
(b) at least one reference expression profile for a particular condition.
18 . (canceled)
19 . A set of miRNAs isolated from a blood sample from a subject for diagnosing and/or prognosing of Wilms' tumour, wherein the miRNAs are selected from the miRNAs as indicated in claim 2 .
20 . The set of miRNAs of claim 19 bound to a carrier, e.g. a micro-array.
21 . Use of a set of miRNAs according to claim 19 for diagnosing and/or prognosing of Wilms' tumour in a subject, particularly a human subject.
22 . The method of claim 1 , wherein said sample is a whole blood sample.