Complex sets of miRNAs as non-invasive biomarkers for kidney cancer
The present invention relates to non-invasive methods, kits and means for diagnosing and/or prognosing of kidney cancer 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 kidney cancer in a body fluid sample from a subject. In addition, the present invention relates to sets of miRNAs for diagnosing and/or prognosing of kidney cancer in a body fluid sample from a subject.
1. A method for diagnosing and/or prognosing of kidney cancer comprising the steps of:
(i) determining an expression profile of a set comprising at least two miRNAs representative for kidney cancer in all blood cells of leukocytes, erythrocytes, and platelets without the acellular portion of a blood sample from a subject, and
(ii) comparing said expression profile to a reference, wherein the comparison of said expression profile to said reference allows for the diagnosis and/or prognosis of kidney cancer,
wherein kidney cancer is a renal cell carcinoma and the nucleotide sequences of the miRNAs comprised in the set are selected from the group consisting of SEQ ID NO: 1 to 465.
2. The method of claim 1 , wherein the set of miRNAs comprises at least one of the sets of miRNAs listed in FIG. 2 or FIG. 9 .
3. The method of claim 1 , wherein the set of miRNAs comprises at least one up-regulated miRNA listed in FIG. 7 a or 7 b and/or at least one down-regulated miRNA listed in FIG. 8 a or 8 b.
4. A kit for diagnosing and/or prognosing of kidney cancer comprising
(i) means for determining an expression profile of a set comprising at least two miRNAs representative for kidney cancer in all blood cells of leukocytes, erythrocytes, and platelets without the acellular portion of a blood sample from a subject comprising:
(a) a set of at least two polynucleotides or set of at least two primer pairs for detecting a set comprising at least two miRNAs for diagnosing and/or prognosing of kidney cancer in a blood sample from a subject, wherein the nucleotide sequences of the miRNAs comprised in the set are selected from the group consisting of SEQ ID NO: 1 to 465,
(b) a biochip, a RT-PCR system, a PCR-system, a flow cytometer, a bead-based multiplex system or a next generation sequencing system,
and
(ii) at least one reference
wherein kidney cancer is a renal cell carcinoma.
5. The method of claim 1 , wherein the expression profile of the miRNA s is determined comprising the steps:
(i) extracting total RNA from all blood cells of leukocytes, erythrocytes, and platelets without the acellular portion of said blood sample,
(ii) reverse-transcribing the total RNA into cDNA, and
(iii) amplifying the cDNA and thereby detecting the miRNA levels in all blood cells of leukocytes, erythrocytes, and platelets without the acellular portion of said blood sample.
6. The kit of claim 4 , wherein the reference is determined in the same type of blood sample as the subject to be diagnosed and/or prognosed.