SiRNA TARGETING ETS1 AND ELK1 AND METHOD OF USING SAME IN THE INHIBITION OF CIP2A GENE IN CANCER TREATMENT

• US 20130225657A1
• Filed: 02/26/2013
• Published: 08/29/2013
• Est. Priority Date: 02/28/2012
• Status: Active Grant

• Current Assignee: Medical Diagnostic Laboratories LLC
• Original Assignee: Medical Diagnostic Laboratories LLC
• Inventors: Rice, Lyndi M., Pallai, Rajash
• Granted Patent: US 9,139,829 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/0053 Mouth and digestive tract, i.e. intraoral and peroral administration rectal administration A61K9/0031
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/31 Combination therapy

Abstract

Disclosed are methods of attenuating activity of the gene promoter of CIP2A. siRNAs are used to target against Ets1 and Elk1 transcriptional factors, thereby blocking the binding of Ets1 and Elk1 to the CIP2A gene promoter. It is disclosed that the siRNAs targeted against Ets1 and Elk1 attenuate the gene expression of CIP2A. A kit containing siRNA reagents for attenuating the CIP2A gene expression is also disclosed.

Claims

1. A method of inhibiting gene expression of CIP2A in a cell, comprising the steps of:

(i) providing a first siRNA targeted against Ets1 mRNA, said first siRNA hybridizes to a target sequence of Ets1 mRNA selected from the group consisting of SEQ ID NO;

34, SEQ ID NO;

35, SEQ ID NO;

36 and SEQ ID NO;

37;

(ii) providing a second siRNA targeted against Elk1 mRNA, said second siRNA hybridizes to a target sequence of Elk1 mRNA selected from the group consisting of SEQ ID NO;

38, SEQ ID NO;

39, SEQ ID NO;

40 and SEQ ID NO;

41; and

(iii) exposing said first siRNA and said second siRNA to a cell, wherein said first siRNA and said second siRNA together inhibit the gene expression of CIP2A in said cell.

2. The method of claim 1, wherein said first siRNA is at least one siRNA selected from the group consisting of SEQ ID NO:

30 and SEQ ID NO;

31.

3. The method of claim 1, wherein said second siRNA is at least one siRNA selected from the group consisting of SEQ ID NO:

32 and SEQ ID NO;

33.

4. A pharmaceutical composition, comprising a first siRNA targeted against Ets1 mRNA;

a second siRNA targeted against Elk1 mRNA; and

a pharmaceutical acceptable carrier, said pharmaceutical composition inhibits gene expression of CIP2A.

5. The pharmaceutical composition of claim 4, wherein said first siRNA contains at least one siRNA selected from the group consist of SEQ ID NO:

30 and SEQ ID NO;

31.

6. The pharmaceutical composition of claim 4, wherein said second siRNA contains at least one siRNA selected from the group consisting of SEQ ID NO:

32 and SEQ ID NO;

33.

7. A kit comprising a packaging material;

and said pharmaceutical composition of claim 4.

8. The kit of claim 7, further comprises an instruction for performing a qPCR to quantify gene expression of CIP2A.

9. An isolated siRNA consisting of the nucleotide sequence of SEQ ID NO:

30 or SEQ ID NO;

31.

11. The isolated siRNA of claim 9, wherein said isolated siRNA comprises a modified nucleotide.

10. An isolated siRNA consisting of the nucleotide sequence of SEQ ID NO:

32 or SEQ ID NO;

33.

12. The isolated siRNA of claim 10, wherein said isolated siRNA comprises a modified nucleotide.

13. A method for inhibiting gene expression of CIP2A in a cell in vitro, said method comprising the steps of:

(a) introducing into a cell a first siRNA, wherein said first siRNA is at least one siRNA selected from the group consisting of SEQ ID NO;

30 and SEQ ID NO;

31;

(b) introducing into said cell a second siRNA, wherein said second siRNA is at least one siRNA selected from the group consisting of SEQ ID NO;

32 and SEQ ID NO;

33; and

(c) culturing said cell to inhibit gene expression of CIP2A, wherein said inhibition of gene expression of CIP2A is evidenced by a decrease in CIP2A mRNA transcript.

14. A method for inhibiting gene expression of CIP2A in a human in vivo, comprising the step of administering to said human an effective amount of a first siRNA targeted Ets1 mRNA and a second siRNA targeted against Elk1 mRNA.

15. The method of claim 14, wherein said first siRNA consists of a nucleotide sequence set forth in SEQ ID NO:

30 or SEQ ID NO;

31.

16. The method of claim 14, wherein said second siRNA consists of a nucleotide sequence set forth in SEQ ID NO:

32 or SEQ ID NO;

33.

17. The method of claim 14, wherein said human is suffering from cervical cancer or endometrial cancer.