Inhibitory RNAs to RNA binding proteins hnRNPA1, hnRNPA2 and PTB and uses thereof

Patent metadata

Field	Value
Publication number	US-9206426-B2
Application number	US-201113582921-A
Country	US
Kind code	B2
Filing date	Mar 7, 2011
Priority date	Mar 5, 2010
Publication date	Dec 8, 2015
Grant date	Dec 8, 2015

Abstract

Provided herein are methods to reduce or slow down cell growth, for example in a cancer cell comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB. Provided are methods to identify agents which reduce the levels of hnRNPA1, hnRNPA2 or PTB. Provided are methods to identify agents which increase ratio of PKM1/PKM2 proteins, or reduce PKM2 levels, or increase PKM1 levels.

First claim

What is claimed is:

1. A method to reduce cell growth of a cancer cell comprising contacting a cancer cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

2. A method to treat cancer comprising administering to a subject in need thereof an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB mRNAs, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

3. A method to treat cancer comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

4. A method to reduce the levels of RNA binding proteins hnRNPA1, hnRNPA2 and PTB in a cell comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

5. A method to reduce the amount of hnRNPA1, hnRNPA2 and PTB protein bound to pyruvate kinase pre-mRNA in a cell, comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

6. A method to reduce levels of PKM2 RNA or protein in a cell, comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

7. A method to increase levels of PKM1 RNA or protein in a cell, comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

8. A method to reduce the ratio of PKM2/PKM1 RNA or protein in a cell, comprising contacting a cell with an effective amount of a combination of inhibitory RNA molecules targeting hnRNPA1, hnRNPA2 and PTB, wherein the inhibitory RNA molecule targeting hnRNPA1 comprises SEQ ID NO: 9 or is encoded by SEQ ID NO: 64, the inhibitory RNA molecule targeting hnRNPA2 comprises SEQ ID NO: 10 or is encoded by SEQ ID NO: 65, and the inhibitory RNA molecule targeting PTB comprises SEQ ID NO: 11 or is encoded by SEQ ID NO: 63.

9. The method of any of claims 1 - 8 , wherein the inhibitory RNA molecule, which targets hnRNPA1, hnRNPA2 or PTB mRNAs is a siRNA.

10. The method of any of claims 1 - 8 , wherein the inhibitory RNA molecule, which targets hnRNPA1, hnRNPA2 or PTB mRNAs is a shRNA.

11. The method of claim 10 , wherein the shRNA is comprised in an RNAi expression vector.

12. The method of any of claims 4 - 8 , wherein the cell is a cancer cell.

13. The method of claim 12 , wherein the cancer cell is brain tumor cell, a glioma tumor cell, or a breast cancer cell.

14. The method of any of claims 4 - 8 wherein the cell expresses PKM2.

15. The method of any of claims 1 - 8 , wherein SEQ ID NOs: 9, 10 and 11 are the sense strand sequence of the inhibitory RNA molecule.

Assignees

• Manley James L
• Chen Mo
• David Charles
• Zhang Jian
• Univ Columbia

Inventors

• Manley James L
• Chen Mo
• David Charles
• Zhang Jian

Classifications

• C12N15/113 Primary

  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 title

• A61K31/7105

  Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N2320/31

  Combination therapy · CPC title

• C12N15/1137 Primary

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title