Treatment of pancreatic developmental gene related diseases by inhibition of natural antisense transcript to a pancreatic developmental gene
View Patent ↗The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of a Pancreatic Developmental gene, in particular, by targeting natural antisense polynucleotides of a Pancreatic Developmental gene. The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of Pancreatic Developmental genes.
1. A method of upregulating a function of and/or the expression of a Pancreatic Developmental gene polynucleotide in patient cells or tissues in vivo or in vitro comprising:
contacting said cells or tissues with at least one single stranded antisense oligonucleotide 19 to 21 nucleotides in length wherein said at least one oligonucleotide has at least 90% sequence identity to a reverse complement of a polynucleotide comprising 19 to 21 consecutive nucleotides within the natural antisense polynucleotide selected from the group consisting of nucleotides 1 to 1235 of SEQ ID SEQ ID NO: 6, 1 to 17,964 of SEQ ID NO: 7, 1 to 1 to 50,003 of SEQ ID SEQ ID NO: 8, 1 to 486 of SEQ ID NO: 9, 1 to 494 of SEQ ID NO: 10, 1 to 1992 of SEQ ID NO: 11, or 1 to 1767 of SEQ ID NO: 12 and wherein said antisense oligonucleotide specifically targets said natural antisense polynucleotide; thereby upregulating a function of and/or the expression of the Pancreatic Developmental gene polynucleotide in patient cells or tissues in vivo or in vitro.
2. A method of upregulating a function of and/or the expression of a Pancreatic Developmental gene polynucleotide selected from the group comprising SEQ ID NOS: 1-5 in patient cells or tissues in vivo or in vitro comprising:
contacting said cells or tissues with at least one single stranded antisense oligonucleotide 19 to 21 nucleotides in length wherein said at least one antisense oligonucleotide has at least 90% sequence identity to a corresponding 19 to 21 nucleotide region within a reverse complement of a natural antisense polynucleotide of a Pancreatic Developmental gene polynucleotide wherein said antisense oligonucleotide specifically targets said natural antisense polynucleotide; thereby upregulating a function of and/or the expression of the Pancreatic Developmental gene polynucleotide in patient cells or tissues in vivo or in vitro.
3. A method of upregulating a function of and/or the expression of a Pancreatic Developmental gene polynucleotide in patient cells or tissues in vivo or in vitro comprising:
contacting said cells or tissues with at least one modified single stranded antisense oligonucleotide of 19 to 21 nucleotides in length that specifically targets a complementary region of a natural antisense polynucleotide the Pancreatic Developmental gene; thereby upregulating a function of and/or the expression of the Pancreatic Developmental gene in patient cells or tissues in vivo or in vitro.
4. The method of claim 3 , wherein a function of and/or the expression of Pancreatic Developmental gene is increased in vitro with respect to a mock-transfected control.
5. The method of claim 3 , wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide selected from the group consisting of SEQ ID NOS: 6-12.
6. The method of claim 3 , wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide antisense to non-coding nucleic acid sequences of a Pancreatic Developmental gene polynucleotide.
7. The method of claim 3 , wherein the at least one antisense oligonucleotide targets a natural antisense polynucleotide having overlapping and/or non-overlapping sequences with a Pancreatic Developmental gene polynucleotide.
8. The method of claim 3 , wherein at least one antisense oligonucleotide comprises one or more modifications selected from: at least one modified sugar moiety, at least one modified internuclcoside linkage, at least one modified nucleotide, and combinations thereof.
9. The method of claim 8 , 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.
10. The method of claim 8 , wherein the one or more modifications comprise at least one modified internucleoside linkage selected from: a phosphorothionate, alkylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, carboxymethyl ester, and combinations thereof.
11. The method of claim 8 , wherein the one or more modifications comprise at least modified nucleotide selected from: a peptide nucleic acid (PNA), a locked nucleic acid (LNA), an arabino-nucleic acid (FANA), and combinations thereof.
12. The method of claim 1 , wherein the at least one oligonucleotide comprises at least one oligonucleotide sequences set forth as SEQ ID NOS: 13 to 45.
13. A method of upregulating a function of and/or the expression of a Pancreatic Developmental gene in mammalian cells or tissues in vivo or in vitro comprising:
contacting said cells or tissues with at least one short interfering RNA (siRNA) oligonucleotide 19 to 21 nucleotides in length, said at least one siRNA oligonucleotide being specific for a natural antisense polynucleotide of a Pancreatic Developmental gene, wherein said at least one siRNA oligonucleotide has at least 90% sequence complementarity to a sequence of at least about nineteen consecutive nucleic acids of the natural antisense polynucleotide; and, upregulating a function of and/or the expression of a Pancreatic Developmental gene in mammalian cells or tissues in vivo or in vitro.
14. The method of claim 13 , wherein said oligonucleotide has sequence complementarity to a sequence of at least about nineteen consecutive nucleic acids of the natural antisense polynucleotide of the Pancreatic Developmental gene polynucleotide.
15. A method of upregulating a function of and/or the expression of a Pancreatic Developmental gene in mammalian cells or tissues in vitro comprising:
contacting said cells or tissues with at least one single stranded modified antisense oligonucleotide of about 19 to 21 nucleotides in length specific for a natural antisense polynucleotide of a Pancreatic Developmental gene wherein said at least one antisense oligonucleotide has at least 90% sequence identity to a 19 to 21 consecutive nucleotide region of the group consisting of SEQ ID NOS: 1 to 5 or polynucleotides transcribed from the Pancreatic Developmental gene; and, upregulating the function and/or expression of the Pancreatic Developmental gene in mammalian cells or tissues in vitro.