Antisense microRNA and uses therefor

• US 7,812,003 B2
• Filed: 08/01/2008
• Issued: 10/12/2010
• Est. Priority Date: 08/02/2007
• Status: Expired due to Fees

• Current Assignee: Stephen H. Safe, Sudhakar Chintharlapalli, Susanne U. Talcott
• Original Assignee: Stephen H. Safe, Sudhakar Chintharlapalli, Susanne U. Talcott
• Inventors: Talcott, Susanne U., Chintharlapalli, Sudhakar, Safe, Stephen H.
• Primary Examiner(s): BOWMAN, AMY HUDSON
• Application Number:
  US12/221,266
• Publication Number:
  US 20090099123A1
• Time in Patent Office: 802 Days
• Field of Search: None
• US Class Current: 514/44.00A
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/113 targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3521 Methyl

Abstract

Provided herein are methods to suppress specificity protein (Sp) activity in a cell associated with a cell proliferative disease. The methods are effective to inhibit a microRNA in the cell using an antisense microRNA oligonucleotide which results in an increase in expression of a specificity protein (Sp) suppressor gene thereby inducing Sp degradation, apoptosis or growth arrest by releasing inhibitors of G2/M (Myt-1) or inhibition. Also provided are methods of treating a cancer using the antisense microRNA oligonucleotide. In addition the present invention provides antisense microRNA-27a oligonucleotides useful in the methods described herein.

Claims

1. A method for suppressing specificity protein (Sp) activity in a breast cancer cell, comprising:

contacting a miR-27a microRNA in the cell with a microRNA antisense oligonucleotide miR-27a consisting of the sequence shown in SEQ ID NO;

1, thereby increasing expression of ZBTB10 gene effective to induce cell cycle arrest in the cell.

2. The method of claim 1, wherein the microRNA antisense oligonucleotide is modified to further comprise a stabilizing group.

3. The method of claim 2, wherein said stabilizing group is a morpholino group.

4. The method of claim 1, wherein the suppressed Sp protein is Sp1, Sp3 or Sp4 or other Sp/KLF protein.

5. The method of claim 1, wherein the gene effective to induce cell cycle arrest is Myt-1.

6. The method of claim 5, wherein Myt-1 induces arrest in G2/M.

7. A method for inhibiting expression of a microRNA in a breast cancer cell, comprising:

delivering to said breast cancer cell, a antisense microRNA oligonucleotide as-microRNA-a27 consisting of the sequence shown in SEQ ID NO;

1.

8. The method of claim 7, wherein the delivery step comprises combining the antisense microRNA with a cationic lipid effective to target the cell.

9. The method of claim 7, wherein the antisense microRNA oligonucleotide is modified to further comprise a stabilizing group.

10. The method of claim 9, wherein said stabilizing group is a morpholino group.

11. A method of treating breast cancer in a subject, comprising:

administering a pharmacologically effective amount of an antisense microRNA-27a oligonucleotide consisting of the sequence shown in SEQ ID NO;

1 to the subject thereby treating the cancer.

12. The method of claim 11, wherein the antisense microRNA oligonucleotide is modified to further a stabilizing group.

13. The method of claim 12, wherein said stabilizing group is a morpholino group.

14. An antisense microRNA-27a oligonucleotide consisting of the sequence shown in SEQ ID NO:

1.

15. The antisense microRNA-27a oligonucleotide of claim 14, wherein the one or more nucleotides are modified with a stabilizing group.

16. The antisense microRNA-27a oligonucleotide of claim 15, wherein the nucleotide(s) is modified with one or more of a morpholino group, a 2′

-O-methyl group or a phosphorothioate derivative.

17. The method of claim 14, wherein one or more nucleotides are modified.