Modulation of gene expression and screening for deregulated protein expression

• US 10,196,639 B2
• Filed: 10/07/2016
• Issued: 02/05/2019
• Est. Priority Date: 10/09/2015
• Status: Active Grant

• Current Assignee: University of Southampton
• Original Assignee: University of Southampton
• Inventors: Vorechovsky, Igor, Kralovicova, Jana
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US15/288,415
• Publication Number:
  US 20170101641A1
• Time in Patent Office: 851 Days
• Field of Search: None
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3519 Fusion with another nucleic acid
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing
• CPC Class Codes: C12N 2320/34 Allele or polymorphism specific uses
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 : Expression markers

Abstract

Disclosed herein include compositions and methods of modulating protein expression that utilizes an activator or a repressor of a non-sense mediated RNA decay switch exon (NSE). In some embodiments, also included herein are compositions and methods of modulating protein expression that uses an agent that targets a transposed element.

Claims

1. A method of modulating protein expression, comprising:

a) contacting a non-sense mediated RNA decay switch exon (NSE)-repressor agent to a target motif within a pre-processed mRNA transcript, wherein the target motif is located in an intronic region between two canonical exons;

b) processing a mRNA transcript from the pre-processed mRNA transcript thereby forming a processed mRNA transcript, wherein the NSE-repressor agent modulates exclusion of an NSE from the processed mRNA transcript; and

c) translating the processed mRNA transcript wherein the exclusion of the NSE upregulates protein expression relative to the protein expression of an equivalent mRNA transcript comprising the NSE.

2. The method of claim 1, wherein the NSE-repressor agent comprises a polynucleic acid polymer that hybridizes to the target motif within the intronic region.

3. The method of claim 2, wherein the polynucleic acid polymer hybridizes to:

a motif within a transposed element, upstream of a transposed element, or downstream of a transposed element;

or a motif within ATM intron 28.

4. The method of claim 3, wherein the transposed element is Alu or MER51.

5. The method of claim 2, wherein the polynucleic acid polymer is from 10 to 50 nucleotides in length.

6. The method of claim 2, wherein the polynucleic acid polymer comprises a sequence with at least 70% sequence identity to a sequence selected from the group consisting of SEQ ID NOs:

23, 26, 37, 39, 51, and 53-55.

7. The method of claim 2, wherein the polynucleic add polymer hybridizes to the NSE within the sequence

8. The method of claim 2, wherein the polynucleic add polymer is modified at a nucleoside moiety, at a phosphate moiety, at a 5′

terminus, at a 3′

terminus, or a combination thereof.

9. The method of claim 2, wherein the polynucleic add polymer comprises an artificial nucleotide selected from the group consisting of 2′

-O-methyl, 2′

-O-methoxyethyl (2′

-O-MOE), 2′

-0-aminopropyl, 2′

-deoxy, T-deoxy-2′

-fluoro, 2′

-O-aminopropyl (2′

-O-AP), 2′

-O-dimethylaminoethyl (2′

-O-DMAOE), 2′

-O-dimethylaminopropyl (2′

-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′

-O-DMAEOE), 2′

-O—

N-methylacetamido (2-O-NMA), a locked nucleic acid (LNA), an ethylene nucleic acid (ENA), a peptide nucleic acid (PNA), a 1′

,5′

-anhydrohexitol nucleic acid (HNA), a morpholino, a methylphosphonate nucleotide, a thiolphosphonate nucleotide, and a 2′

-fluoro N3-P5′

-phosphoramidite.

10. The method of claim 1, wherein the pre-processed mRNA transcript is an endogenous pre-processed mRNA transcript.

11. The method of claim 1, wherein a protein translated from the processed mRNA is:

a wild-type protein;

or

a full-length protein.

17. The method of claim 1, wherein the NSE-repressor agent modulates exclusion of a portion of the NSE from the processed mRNA transcript.

18. The method of claim 1, wherein the NSE-repressor agent promotes exclusion of the NSE from the processed mRNA transcript.

12. A method of treating or preventing a disease or condition in a subject in need thereof, the method comprising:

administering to the subject a pharmaceutical composition comprising;

(i) a non-sense mediated RNA decay switch exon (NSE)-repressor agent that interacts with a target motif within a pre-processed mRNA transcript to modulate exclusion of an NSE from a processed mRNA transcript wherein the target motif is located in an intronic region between two canonical exons;

and

(ii) a pharmaceutically acceptable excipient and/or a delivery vehicle;

wherein the disease or condition is treated or prevented in the subject by the administration of the NSE-repressor agent by an increase in expression of a protein translated from the processed mRNA transcript.

13. The method of claim 12, wherein the disease or condition is cancer.

14. The method of claim 12, wherein the NSE-repressor agent comprises a polynucleic acid polymer that hybridizes to the target motif within the intronic region.

15. The method of claim 14, wherein the polynucleic acid polymer hybridizes to the NSE within the sequence

16. The method of claim 12, wherein a protein translated from the processed mRNA is a full-length protein.

19. The method of claim 12, wherein the NSE-repressor agent modulates exclusion of a portion of the NSE from the processed mRNA transcript.

20. The method of claim 12, wherein the NSE-repressor agent promotes exclusion of the NSE from the processed mRNA transcript.