Antisense modulation of human Rho family gene expression

Abstract

This invention provides compositions and methods for modulating expression of members of the human Rho gene family, which encode low molecular weight GTPases that act as molecular switches in signal transduction. In preferred embodiments, Rho family members include RhoA, RhoB, RhoC, RhoG, Rac1 and cdc42. This invention is also directed to methods for inhibiting hyperproliferation of cells; these methods can be used diagnostically or therapeutically. Furthermore, this invention is directed to treatment of conditions associated with expression of the human Rho family members, particularly in hyperproliferative disorders.

What is claimed is:  
     
         1 . An antisense compound targeted to a nucleic acid molecule encoding a member of the human Rho family of small GTP binding proteins, wherein said antisense compound inhibits the expression of said member of the human Rho family.  
     
     
         2 . The antisense compound of  claim 1  which is an antisense oligonucleotide.  
     
     
         3 . The antisense compound of  claim 2  wherein the oligonucleotide comprises at least one modified internucleoside linkage.  
     
     
         4 . The antisense compound of  claim 3  wherein the modified internucleoside linkage is a phosphorothioate linkage.  
     
     
         5 . The antisense compound of  claim 2  wherein the oligonucleotide comprises at least one modified sugar moiety.  
     
     
         6 . The antisense compound of  claim 5  wherein the modified sugar moiety is a 2′-O-methoxyethyl sugar moiety.  
     
     
         7 . The antisense compound of  claim 2  wherein the oligonucleotide comprises at least one modified nucleobase.  
     
     
         8 . The antisense compound of  claim 7  wherein the modified nucleobase is a 5-methylcytosine.  
     
     
         9 . The antisense compound of  claim 2  wherein the oligonucleotide is a chimeric oligonucleotide.  
     
     
         10 . The antisense compound of  claim 1  which is from 8 to 30 nucleobases in length.  
     
     
         11 . The antisense compound of  claim 1  wherein said member of the human Rho family of small GTP binding proteins is RhoA, RhoB, RhoC, or RhoG.  
     
     
         12 . The antisense compound of  claim 1  wherein said member of the human Rho family of small GTP binding protein is rac1.  
     
     
         13 . The antisense compound of  claim 12  wherein the oligonucleotide comprises SEQ ID NO: 193, 195, 199, 201, 204 or 205.  
     
     
         14 . The antisense compound of  claim 1  wherein said member of the human Rho family of small GTP binding protein is cdc42.  
     
     
         15 . The antisense compound of  claim 14  wherein the oligonucleotide comprises SEQ ID NO: 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229 or 230.  
     
     
         16 . A pharmaceutical composition comprising the antisense compound of  claim 1  and a pharmaceutically acceptable carrier or diluent.  
     
     
         17 . The pharmaceutical composition of  claim 16  further comprising a colloidal dispersion system.  
     
     
         18 . The pharmaceutical composition of  claim 16  wherein the antisense compound is an antisense oligonucleotide.  
     
     
         19 . A method of inhibiting the expression of a member of the human Rho family of small GTP binding proteins in human cells or tissues comprising contacting said cells or tissues with the antisense compound of  claim 1  so that expression of said human Rho family member is inhibited.  
     
     
         20 . A method of treating a human having a disease or condition associated with a member of the human Rho family of small GTP binding proteins comprising administering to said animal a therapeutically or prophylactically effective amount of the antisense compound of  claim 1  so that expression of said human Rho family member is inhibited.  
     
     
         21 . The method of  claim 20  wherein the disease or condition is a hyperproliferative condition.  
     
     
         22 . The method of  claim 21  wherein the hyperproliferative condition is cancer.  
     
     
         23 . The method of  claim 20  wherein the disease or condition is abnormal wound healing or clotting.  
     
     
         24 . The method of  claim 20  wherein the disease or condition is ischemia/reperfusion or reoxygenation injury.  
     
     
         25 . A compound which inhibits JNK activation by a non-cytokine activator but does not inhibit JNK activation by a cytokine.  
     
     
         26 . The compound of  claim 25  wherein the non-cytokine activator is a stress signal.  
     
     
         27 . The compound of  claim 26  wherein the stress signal is hydrogen peroxide or ultraviolet radiation.  
     
     
         28 . The compound of  claim 25  wherein the cytokine is IL-1β.  
     
     
         29 . The compound of  claim 25  which is an inhibitor of rhoA.  
     
     
         30 . The compound of  claim 29  which is an antisense inhibitor of rhoA.  
     
     
         31 . A method of inhibiting JNK activation by a non-cytokine activator without inhibiting JNK activation by a cytokine comprising contacting JNK or cells or tissues containing JNK with an inhibitor of rhoA.  
     
     
         32 . The method of  claim 31  wherein the non-cytokine activator is a stress signal.  
     
     
         33 . The method of  claim 32  wherein the stress signal is hydrogen peroxide or ultraviolet radiation.  
     
     
         34 . The method of  claim 31  wherein the cytokine is IL-1β.  
     
     
         35 . The method of  claim 31  wherein the Inhibitor of rhoA is an antisense oligonucleotide inhibitor of rhoA.