Non-invasive method for the diagnosis or screening of colorectal cancer and/or pre-cancerous stage thereof

1 . An in vitro method for diagnosis or screening colorectal cancer or a pre-cancerous advanced colorectal adenoma in a subject at risk, comprising: a) measuring the expression level of at least miR-421 in a biological sample obtained from the subject at risk, wherein the biological sample is a stool sample; (b) identifying the subject at risk as being a subject suffering from colorectal cancer and/or a pre-cancerous advanced colorectal adenoma based on a risk score that is generated by comparing the expression level of the at least miR-421 to a reference expression level as measured in healthy control subjects, wherein a positive risk score is obtained from a finding of an overexpression of at least the miRNA-421; and (c) subjects having a positive risk score from step (b), conducting an imaging technique on the subject suffering from colorectal cancer and/or a pre-cancerous advanced colorectal adenoma to confirm a positive diagnosis of colorectal cancer or a pre-cancerous advanced colorectal adenoma.

2 . The in vitro method, according to the claim 1 , which comprises: a) measuring the expression level of at least one combination of [miR-421 and miR-27a], [miR-421 and miR-25], [miR-421 and miR-221], [miR-421 and miR-34a], [miR-421 and miR-29a] or [miR-421 and miR-130b] in the biological sample; (b) identifying the subject at risk as being a subject suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma based on a risk score that is generated by comparing the expression level of the at least one combination to reference expression levels as measured in healthy control subjects and wherein a positive risk score is obtained from a finding of an overexpression of the at least one combination.

3 . The in vitro method, according to claim 1 , wherein the subject at risk is from 50 to 75-years-old.

4 . The in vitro method, according to claim 1 , which further comprises determining the presence or concentration of haemoglobin, wherein the presence or higher concentration of haemoglobin as compared with healthy control subjects is indicative that the subject at risk is suffering from colorectal cancer or pre-cancerous advanced colorectal adenoma.

5 . The in vitro method, according to claim 1 , wherein the stool sample is less than 5 mg.

6 . The in vitro method, according to claim 1 , wherein the measuring of the expression level of the at least miRNA-421 is carried out in a stool sample which has been previously used for the determination of the presence or concentration of haemoglobin.

7 . The in vitro method, according to claim 1 , wherein the imaging technique is a colonoscopy.

8 . The in vitro method according claim 1 , which further comprises determining the presence or concentration of haemoglobin, wherein the presence or higher concentration of haemoglobin as compared with healthy control subjects, is indicative that the subject at risk is suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma, and wherein the stool sample is less than 5 mg.

9 . The method of claim 1 , further comprising collecting a stool sample prior to measuring the expression level.

10 . The method of claim 1 , further comprising treating the subject for a cancer or for a pre-cancerous lesion by endoscopic polypectomy, surgical resection, chemotherapy, or radiotherapy.

11 . The in vitro method, according to claim 5 , wherein the stool sample is less than 2.5 mg.

12 . An in vitro method for diagnosis or screening colorectal cancer or a pre-cancerous advanced colorectal adenoma in a subject at risk, comprising (a) collecting a stool sample from the subject at risk; (b) measuring the expression level of at least miR-421 in the stool sample; (c) identifying the subject at risk as being a subject suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma based on a risk score that is generated by comparing the expression level of the at least miR-421 to a reference expression level as measured in healthy control subjects, and wherein a positive risk score is obtained from a finding of an overexpression of at least the miRNA-421; and (d) conducting an imaging technique on the subject suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma to confirm a positive diagnosis of colorectal cancer or a pre-cancerous advanced colorectal adenoma.

13 . The in vitro method of claim 12 , further comprising determining the presence or concentration of haemoglobin in the stool sample, wherein when both haemoglobin is present or is present at an increased concentration as compared with healthy control subjects and the miR-421 is overexpressed as compared with healthy control indicates that the subject at risk is suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma.

14 . The in vitro method, according to the claim 12 , which comprises: (a) measuring the expression level of at least one combination of [miR-421 and miR-27a], [miR-421 and miR-25], [miR-421 and miR-221], [miR-421 and miR-34a], [miR-421 and miR-29a] or [miR-421 and miR-130b] in the stool sample, and (b) identifying the subject at risk as being a subject suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma based on a risk score that is generated by comparing processing the expression level of the at least one combination to reference expression levels as measured in healthy control subjects and wherein a positive risk score is obtained from a finding of an overexpression of the at least the one combination.

15 . The in vitro method of claim 14 , further comprising determining the presence or concentration of haemoglobin in the stool sample, wherein when both haemoglobin is present or is present at an increased concentration as compared with healthy control subjects and the at least one combination of miRNAs is overexpressed as compared with healthy control subjects indicates that the subject at risk is suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma.

16 . The in vitro method of claim 10 , further comprising determining the presence or concentration of haemoglobin in the stool sample, wherein when both haemoglobin is present or is present at an increased concentration as compared with healthy control subjects and the miR-421 is overexpressed as compared with healthy control indicates that the subject at risk is suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma.

17 . The in vitro method, according to the claim 10 , which comprises: (a) measuring the expression level of at least one combination of [miR-421 and miR-27a], [miR-421 and miR-25], [miR-421 and miR-221], [miR-421 and miR-34a], [miR-421 and miR-29a] or [miR-421 and miR-130b] in the stool sample, and (b) identifying the subject at risk as suffering from colorectal cancer or a pre-cancerous advanced colorectal adenoma based on a risk score that is generated by comparing the expression level of the at least one combination to reference expression levels as measured in healthy control subjects and wherein a positive risk score is obtained from a finding of an over expression of the at least one combination.