EpCAM aptamer for detection of cancer stem cells
The present disclosure relates to an RNA aptamer and uses thereof, in particular, an aptamer which specifically bind to EpCAM and which demonstrates superior tumor penetrating ability.
1. An RNA aptamer which binds specifically to EpCAM, wherein:
the aptamer is not DT3 having the sequence GCGACUGGUUACCCGGUCG (SEQ ID No:1); and
the aptamer comprises the sequence ACGUAUCCCUUUUCGCGUA (SEQ ID No:2) or a sequence comprising one or more substitutions within the sequence of SEQ ID No:2.
2. The aptamer according to claim 1 wherein the aptamer comprises a sequence length between 19 and 100 bases.
3. The aptamer according to claim 1 wherein the aptamer comprises one or more modifications that increase aptamer stability.
4. The aptamer according to claim 1 , which consists of the sequence of SEQ ID No:2.
5. The aptamer according to claim 1 , wherein the aptamer specifically binds to EpCAM+ cancer stem cell(s).
6. The aptamer according to claim 5 , wherein the cell is a breast cancer stem cell, a prostate cancer stem cell, a pancreatic cancer stem cell, a colon cancer stem cell, a liver cancer stem cell, a lung cancer stem cell, an ovarian cancer stem cell, or a head and neck cancer stem cell.
7. The aptamer according to claim 5 , wherein the cell is present in a biological sample obtained from a subject.
8. A diagnostic agent comprising the RNA aptamer according to claim 1 coupled to a detectable label.
9. The diagnostic agent according to claim 8 , wherein the detectable label is selected from an enzyme, prosthetic group, fluorescent material, luminescent material, bioluminescent material, electron dense label, labels for MRI and radioactive material and combinations thereof.
10. The aptamer according to claim 1 , or a diagnostic agent comprising the aptamer of claim 1 coupled to a detectable label for use in histological examination of biological samples.
11. An anticancer agent comprising the RNA aptamer according to claim 1 coupled to a moiety.
12. The anticancer agent according to claim 11 wherein the moiety is selected from a toxin, a radionuclide, a chemotherapeutic agent and combinations thereof.
13. A method for isolating, purifying or enriching a EpCAM expressing cell(s) and/or cancer stem cell(s) from a biological sample obtained from a subject, the method comprising contacting the cell with an RNA aptamer according to claim 1 or a diagnostic agent comprising the aptamer of claim 1 coupled to a detectable label.
14. A method for identifying a EpCAM expressing cell(s) and/or cancer stem cell(s) in a subject or in a biological sample obtained from a subject having, or suspected of having cancer, the method comprising contacting the cell with an isolated RNA aptamer according to claim 1 or a diagnostic agent comprising the aptamer of claim 1 coupled to a detectable label.
15. The method according to claim 13 , wherein the subject is one which has a cancer selected from breast cancer, prostate cancer, pancreatic cancer, colon cancer, liver cancer, lung cancer, ovarian cancer, head and neck cancer, melanoma or any other cancer in which EpCAM + cells are present.
16. The method according to claim 14 , wherein the subject is one which has a cancer selected from breast cancer, prostate cancer, pancreatic cancer, colon cancer, liver cancer, lung cancer, ovarian cancer, head and neck cancer, melanoma or any other cancer in which EpCAM + cells are present.
17. A method for treating or preventing cancer in a subject comprising providing a subject with an RNA aptamer according to claim 1 , or an anticancer agent comprising the RNA aptamer according to claim 1 coupled to a moiety.
18. A delivery agent comprising an RNA aptamer according to claim 1 , coupled to an siRNA or ribozyme.
19. The delivery agent according to claim 18 comprising the sequence ACGUAUCCCUUUUCGCGUAAAAUGUAGAGAUGCGGUGGUCCUU (SEQ ID NO:3).
20. A composition comprising:
one of:
a therapeutically effective amount of an RNA aptamer comprising the sequence of SEQ ID No:2 wherein the aptamer binds specifically to EpCAM;
an anticancer agent comprising the RNA aptamer coupled to a moiety; or
a delivery agent comprising the RNA aptamer coupled to an siRNA or ribozyme;
and a pharmaceutically acceptable carrier and/or excipient.
21. The aptamer of claim 1 , comprising at least one, two, three, four, five or six substitutions within the sequence of SEQ ID No:2.
22. The aptamer of claim 3 , wherein the pyrimidine bases of the aptamer are 2′-fluoro (2′-F) modified.
23. The aptamer of claim 3 , wherein the 3′ end of the aptamer is modified to protect it from nuclease digestion.
24. The aptamer of claim 3 , wherein the aptamer is modified by coupling the 5′ end to a fluorophore, inverted dT or a PEG molecule.