Antisense modulation of interleukin-5 signal transduction

Abstract

Compositions and methods are provided for antisense modulation of interleukin-5 signal transduction. Antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding interleukin-5 and interleukin-5 receptor a are preferred. Methods of using these compounds for modulation of interleukin-5 signal transduction and for treatment of diseases associated with interleukin-5 signal transduction are also provided.

What is claimed is:  
     
         1 . An antisense compound 8 to 30 nucleobases in length which modulates interleukin-5 signal transduction.  
     
     
         2 . The antisense compound of  claim 1  which is an antisense oligonucleotide.  
     
     
         3 . The antisense compound of  claim 1  which is targeted to a nucleic acid molecule encoding mammalian interleukin-5, wherein said antisense compound modulates the expression of mammalian interleukin-5.  
     
     
         4 . An antisense compound up to 30 nucleobases in length comprising at least an 8-nucleobase portion of SEQ ID NO: 52, 53 or 62 which inhibits the expression of mammalian interleukin-5.  
     
     
         5 . The antisense compound of  claim 1  which is targeted to a nucleic acid molecule encoding a mammalian interleukin-5 receptor a, wherein said antisense compound modulates the expression of mammalian interleukin-5 receptor a.  
     
     
         6 . An antisense compound up to 30 nucleobases in length comprising at least an 8-nucleobase portion of SEQ ID NO: 162, 166, 167, 169, 170, 171 or 172 which inhibits the expression of mammalian interleukin-5 receptor a.  
     
     
         7 . The antisense compound of  claim 2  wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.  
     
     
         8 . The antisense compound of  claim 7  wherein the modified internucleoside linkage of the antisense oligonucleotide is a phosphorothioate linkage.  
     
     
         9 . The antisense compound of  claim 7  wherein the modified internucleoside linkage of the antisense oligonucleotide is a peptide nucleic acid.  
     
     
         10 . The antisense compound of  claim 9  which comprises at least one basic amino acid conjugated to at least one end of the antisense compound.  
     
     
         11 . The antisense compound of  claim 10  wherein the basic amino acid is lysine or arginine.  
     
     
         12 . The antisense compound of  claim 10  which is less than 20 nucleobases in length.  
     
     
         13 . The antisense compound of  claim 12  comprising at least an 8-nucleobase portion of SEQ. ID NO: 209.  
     
     
         14 . The antisense compound of  claim 2  wherein the antisense oligonucleotide comprises at least one modified sugar moiety.  
     
     
         15 . The antisense compound of  claim 14  wherein the modified sugar moiety of the antisense oligonucleotide is a 2′-O-methoxyethyl sugar moiety.  
     
     
         16 . The antisense compound of  claim 15  wherein substantially all sugar moieties of the antisense oligonucleotide are 2′-O-methoxyethyl sugar moieties.  
     
     
         17 . The antisense compound of  claim 2  wherein the antisense oligonucleotide comprises at least one modified nucleobase.  
     
     
         18 . The antisense compound of  claim 17  wherein the modified nucleobase of the antisense oligonucleotide is a 5-methylcytosine.  
     
     
         19 . The antisense compound of  claim 15  wherein each 2′-O-methoxyethyl modified cytosine nucleobase of the antisense oligonucleotide is a 5-methylcytosine.  
     
     
         20 . The antisense compound of  claim 1  which is a chimeric oligonucleotide.  
     
     
         21 . A pharmaceutical composition comprising the antisense compound of  claim 1  and a pharmaceutically acceptable carrier or diluent.  
     
     
         22 . The pharmaceutical composition of  claim 21  further comprising a colloidal dispersion system.  
     
     
         23 . The pharmaceutical composition of  claim 21  wherein the antisense compound is an antisense oligonucleotide.  
     
     
         24 . The antisense compound of  claim 5  which is targeted to soluble interleukin-5 receptor a and which preferentially inhibits the expression of soluble interleukin-5 receptor a.  
     
     
         25 . The antisense compound of  claim 24  which is targeted to a region of a nucleic acid molecule encoding soluble interleukin-5 receptor a which is not found in a nucleic acid molecule encoding membrane interleukin-5 receptor a.  
     
     
         26 . The antisense compound of  claim 5  which is targeted to membrane interleukin-5 receptor a and which preferentially inhibits the expression of membrane interleukin-5 receptor a.  
     
     
         27 . The antisense compound of  claim 26  which is targeted to a region of a nucleic acid molecule encoding membrane interleukin-5 receptor a which is not found in a nucleic acid molecule encoding soluble interleukin-5 receptor a.  
     
     
         28 . The antisense compound of  claim 5  which inhibits the expression of both soluble and membrane forms of interleukin-5 receptor a.  
     
     
         29 . The antisense compound of  claim 5  which alters the ratio of interleukin-5 receptor a isoforms expressed by a cell or tissue.  
     
     
         30 . The antisense compound of  claim 29  which increases the ratio of the soluble form of interleukin-5 receptor a to the membrane form of interleukin-5 receptor a expressed.  
     
     
         31 . The antisense compound of  claim 30  which is an antisense oligonucleotide wherein substantially all sugar moieties of the antisense oligonucleotide are 2′-O-methoxyethyl sugar moieties.  
     
     
         32 . The antisense compound of  claim 5  which promotes apoptosis.  
     
     
         33 . An antisense compound which alters splicing of an RNA encoding interleukin-5 receptor a, such that the ratio of interleukin-5 receptor a isoforms is altered.  
     
     
         34 . The antisense compound of  claim 33  wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.  
     
     
         35 . The antisense compound of  claim 34  wherein the modified internucleoside linkage of the antisense oligonucleotide is a phosphorothioate linkage.  
     
     
         36 . The antisense compound of  claim 34  wherein the modified internucleoside linkage is a peptide nucleic acid.  
     
     
         37 . The antisense compound of  claim 36  which comprises at least one basic amino acid conjugated to at least one end of the antisense compound.  
     
     
         38 . The antisense compound of  claim 37  wherein the basic amino acid is lysine or arginine.  
     
     
         39 . The antisense compound of  claim 38  which is less than 20 nucleobases in length.  
     
     
         40 . The antisense compound of  claim 36  comprising at least an 8-nucleobase portion of SEQ. ID NO: 209.  
     
     
         41 . The antisense compound of  claim 33  wherein the antisense oligonucleotide comprises at least one modified sugar moiety.  
     
     
         42 . The antisense compound of  claim 41  wherein the modified sugar moiety of the antisense oligonucleotide is a 2′-O-methoxyethyl sugar moiety.  
     
     
         43 . The antisense compound of  claim 42  wherein substantially all sugar moieties of the antisense oligonucleotide are 2′-O-methoxyethyl sugar moieties.  
     
     
         44 . The antisense compound of  claim 33  wherein the antisense oligonucleotide comprises at least one modified nucleobase.  
     
     
         45 . The antisense compound of  claim 44  wherein the modified nucleobase of the antisense oligonucleotide is a 5-methylcytosine.  
     
     
         46 . The antisense compound of  claim 42  wherein each 2′-O-methoxyethyl modified cytosine nucleobase of the antisense oligonucleotide is a 5-methylcytosine.  
     
     
         47 . The antisense compound of  claim 33  which comprises a conjugate group of at least one lysine or arginine linked to the antisense compound.  
     
     
         48 . The antisense compound of  claim 33  which is a chimeric oligonucleotide.  
     
     
         49 . A method of modulating interleukin-5 signal transduction in cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 1  so that interleukin-5 signal transduction is modulated.  
     
     
         50 . A method of modulating the expression of mammalian interleukin-5 in mammalian cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 3  so that expression of mammalian interleukin-5 is inhibited.  
     
     
         51 . A method of modulating the expression of mammalian interleukin-5 receptor a in mammalian cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 33  so that expression of mammalian interleukin-5 receptor a is inhibited.  
     
     
         52 . A method of altering the ratio of the isoforms of mammalian interleukin-5 receptor a in mammalian cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 33  so that the ratio of the mammalian interleukin-5 receptor a isoforms is altered.  
     
     
         53 . A method of modulating the expression of mammalian interleukin-5 receptor a in mammalia cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 5  so that expression of mammalian interleukin-5 receptor a is inhibited.  
     
     
         54 . A method of altering the ratio of the isoforms of mammalian interleukin-5 receptor a in mammalian cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 31  so that the ratio of the mammalian interleukin-5 receptor a isoforms is altered.  
     
     
         55 . A method of treating a mammalian having a disease or condition associated with interleukin-5 signal transduction comprising administering to said mammal a therapeutically or prophylactically effective amount of the antisense compound of  claim 1  so that interleukin-5 signal transduction is modulated.  
     
     
         56 . A method of treating a mammal having a disease or condition associated with interleukin-5 expression comprising administering to said mammal a therapeutically or prophylactically effective amount of the antisense compound of  claim 3  so that interleukin-5 expression is modulated.  
     
     
         57 . A method of treating a mammal having a disease or condition associated with interleukin-5 receptor a expression comprising administering to said mammal a therapeutically or prophylactically effective amount of the antisense compound of  claim 5  so that interleukin-5 receptor a expression is modulated.  
     
     
         58 . The method of  claim 57  wherein the disease or condition is an eosinophilic syndrome or asthma.  
     
     
         59 . The method of  claim 57  wherein the route of administration is pulmonary administration.  
     
     
         60 . A method of treating a mammal having a disease or condition associated with interleukin-5 receptor a expression comprising administering to said mammal a therapeutically or prophylactically effective amount of the antisense compound of  claim 33  so that the ratio of interleukin-5 receptor a isoforms is altered.  
     
     
         61 . The method of  claim 60  wherein the disease or condition is an eosinophilic syndrome or asthma.  
     
     
         62 . The method of  claim 60  wherein the route of administration is pulmonary administration.  
     
     
         63 . A method of treating a mammal having a disease or condition characterized by a reduction in apoptosis comprising administering to said mammal a prophylactically or therapeutically effective amount of the antisense compound of  claim 32 .  
     
     
         64 . A method of treating a mammal having a disease or condition associated with interleukin-5 receptor a expression comprising administering to said mammal a therapeutically or prophylactically effective amount of the antisense compound of  claim 26  so that expression of membrane interleukin-5 receptor a is modulated.  
     
     
         65 . The method of  claim 64  wherein the disease or condition is asthma or an eosinophilic syndrome.  
     
     
         66 . The method of  claim 64  wherein the route of administration is pulmonary administration.  
     
     
         67 . The pharmaceutical composition of  claim 21  further comprising a chemotherapeutic agent for the treatment of asthma.  
     
     
         68 . A pharmaceutical composition comprising the antisense compound of  claim 28  and a pharmaceutically acceptable carrier or diluent.  
     
     
         69 . A pharmaceutical composition comprising the antisense compound of  claim 36  and a pharmaceutically acceptable carrier or diluent.  
     
     
         70 . A diagnostic kit for detecting the expression level of the membrane versus soluble form of IL-5 Receptor a.  
     
     
         71 . The diagnostic kit of claim  70  comprising the antisense compound of  claim 33 .  
     
     
         72 . The diagnostic kit of claim  71  wherein the antisense compound is a peptide nucleic acid.