Treatment of interferon regulatory factor 8 (IRF8) related diseases by inhibition of natural antisense transcript to IRF8

• US 8,946,181 B2
• Filed: 01/04/2011
• Issued: 02/03/2015
• Est. Priority Date: 01/04/2010
• Status: Active Grant

• Current Assignee: CURNA, Inc. (Opko Health, Inc.)
• Original Assignee: CURNA, Inc. (Opko Health, Inc.)
• Inventors: Collard, Joseph, Sherman, Olga Khorkova
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US13/519,529
• Publication Number:
  US 20120295954A1
• Time in Patent Office: 1,491 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/178 miRNA, siRNA or ncRNA

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Interferon Regulatory Factor 8 (IRF8), in particular, by targeting natural antisense polynucleotides of Interferon Regulatory Factor 8 (IRF8). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of IRF8.

Claims

1. A method of upregulating a function of and/or the expression of a Interferon Regulatory Factor 8 (IRF8) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one modified antisense oligonucleotide 12 to 30 nucleotides in length wherein said at least one oligonucleotide is specific for and targets a natural antisense polynucleotide of the IRF8 gene and has identity to a reverse complement of a polynucleotide comprising 12 to 30 consecutive nucleotides within nucleotides 1 to 312 of SEQ ID NO;

2 thereby upregulating a function of and/or the expression of the Interferon Regulatory Factor 8 (IRF8) polynucleotide in patient cells or tissues in vivo or in vitro.

10. The method of claim 1, wherein the at least one oligonucleotide comprises at least one oligonucleotide sequences set forth as SEQ ID NOS:

3 to 6.

2. A method of upregulating a function of and/or the expression of a Interferon Regulatory Factor 8 (IRF8) polynucleotide in patient cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one single stranded modified antisense oligonucleotide of 12 to 30 nucleotides in length that specifically targets a complementary region of a natural antisense polynucleotide of the Interferon Regulatory Factor 8 (IRF8) polynucleotide selected from SEQ ID NO 2;

thereby upregulating a function of and/or the expression of the Interferon Regulatory Factor 8 (IRF8) polynucleotide in patient cells or tissues in vivo or in vitro.

3. The method of claim 2, wherein a function of and/or the expression of the Interferon Regulatory Factor 8 (IRF8) is increased in vivo or in vitro with respect to a control.

4. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to coding nucleic acid sequences of a Interferon Regulatory Factor 8 (IRF8) polynucleotide.

5. The method of claim 2, wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping sequences with a Interferon Regulatory Factor 8 (IRF8) polynucleotide.

6. The method of claim 2, wherein the at least one antisense oligonucleotide comprises one or more modifications selected from:

at least one modified sugar moiety, at least one modified internucleoside linkage, at least one modified nucleotide, and combinations thereof.

7. The method of claim 6, wherein the one or more modifications comprise at least one modified sugar moiety selected from:

a 2′

-O-methoxyethyl modified sugar moiety, a 2′

-methoxy modified sugar moiety, a 2′

-O-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof.

8. The method of claim 6, wherein the one or more modifications comprise at least one modified internucleoside linkage selected from:

a phosphorothioate, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.

9. The method of claim 6, wherein the one or more modifications comprise at least one modified nucleotide selected from:

a peptide nucleic acid (PNA), a locked nucleic acid (LNA), an arabino-nucleic acid (FANA), an analogue, a derivative, and combinations thereof.

11. A method of upregulating a function of and/or the expression of Interferon Regulatory Factor 8 (IRF8) in mammalian cells or tissues in vivo or in vitro comprising:

contacting said cells or tissues with at least one modified antisense oligonucleotide of about 12 to 30 nucleotides in length specific for noncoding sequences of a natural antisense strand of a Interferon Regulatory Factor 8 (IRF8) polynucleotide wherein said at least one antisense oligonucleotide has sequence identity to at least one nucleic acid sequence set forth as SEQ ID NOS;

1 or an RNA polynucleotide encoded by the IRF8 gene; and

, upregulating the function and/or expression of the Interferon Regulatory Factor 8 (IRF8) in mammalian cells or tissues in vivo or in vitro.

12. A method of treating a disease associated with at least one Interferon Regulatory Factor 8 (IRF8) polynucleotide and/or at least one encoded product thereof, comprising:

administering to a patient a therapeutically effective dose of at least one modified antisense oligonucleotide of 12 to 30 nucleotides in length that specifically binds to a natural antisense sequence of said at least one Interferon Regulatory Factor 8 (IRF8) polynucleotide and upregulates expression of said at least one Interferon Regulatory Factor 8 (IRF8) polynucleotide;

thereby treating the disease associated with the at least one Interferon Regulatory Factor 8 (IRF8) polynucleotide and/or at least one encoded product thereof.

13. The method of claim 12, wherein a disease associated with the at least one Interferon Regulatory Factor 8 (IRF8) polynucleotide is selected from a disease or disorder associated with abnormal function and/or expression of IRF8, cancer, a myeloproliferative disorder, Chronic myelogenous leukemia (CML), multiple myeloma, a bone development/metabolic disease or disorder, periodontitis and rheumatoid arthritis, osteoporosis, multiple sclerosis, an immunological disease or disorder, an autoimmune disease or disorder, an immunodeficiency disease or disorder, AIDS, a disease or disorder involving defective innate immunity and a disease associated with apoptosis, aging and senescence.