TREATMENT OF CNS INJURY WITH RNAi THERAPEUTICS

Abstract

Self-delivering PTEN RNA and methods of reducing PTEN expression are provided herein. Also provided are methods of treating spinal cord injury (SCI) and other neurotrauma with PTEN sdRNA.

1 . A phosphatase and tensin homolog (PTEN) targeting agent comprising an isolated sdRNA molecule. 
     
     
         2 . The isolated sdRNA molecule of  claim 1  which comprises a nucleotide sequence complementary to a PTEN gene,
 the isolated sdRNA molecule comprising a guide nucleotide strand, a passenger nucleotide strand, and a cholesterol-TEG molecule attached at the 3′ end of the passenger strand, 
 the isolated sdRNA molecule having a double-stranded region and a single-stranded region,
 the double-stranded region being from 8-15 nucleotides long, and 
 the single-stranded region being at the 3′ end of the guide nucleotide strand, being 4-12 nucleotides long, 
 
 wherein at least 40% of the nucleotides of the sdRNA molecule are modified with at least one modification, and 
 wherein the sdRNA molecule does not form a hairpin. 
 
     
     
         3 . The isolated sdRNA molecule of  claim 2 , wherein the at least one modification is a phosphorothioate, O-methyl, and/or 2-fluror modification. 
     
     
         4 . The sdRNA molecule of  claim 2 , wherein the guide strand has a nucleotide sequence comprising SED ID NO:5 or SEQ ID NO:7. 
     
     
         5 . The sdRNA molecule of  claim 4 , wherein the passenger stand has a nucleotide sequence comprising SEQ ID NO:6 or SEQ ID NO:8. 
     
     
         6 . The sdRNA molecule of  claim 5  which is BA-434 and comprises a guide sequence comprising SEQ ID NO:6 and a passenger sequence comprising SDE ID NO:7. 
     
     
         7 . A pharmaceutical composition comprising the sdRNA molecule of  claim 6  and a pharmaceutically acceptable carrier. 
     
     
         8 . A method of inhibiting PTEN expression in a mammalian cell, comprising contacting the cell with an amount of PTEN sdRNA such that PTEN mRNA expression is inhibited. 
     
     
         9 . The method of  claim 8 , wherein the sdRNA molecule comprises a nucleotide sequence complementary to a PTEN gene,
 the sdRNA molecule comprising a guide nucleotide strand, a passenger nucleotide strand, and a cholesterol-TEG molecule at the 3′ end of the passenger nucleotide strands,   the sdRNA molecule having a double-stranded region and a single-stranded region,
 the double-stranded region being from 8-15 nucleotides long, and 
 the single-stranded region being at the 3′ end of the guide strand, being 4-12 nucleotides long, 
   wherein at least 40% of the nucleotides of the isolated sdRNA molecule are modified with at least one modification, and   wherein the isolated sdRNA molecule does not form a hairpin.   
     
     
         10 . The method of  claim 9 , wherein the sdRNA molecule is BA-434 and comprises a guide sequence comprising SEQ ID NO:8 and a passenger sequence comprising SEQ ID NO:7. 
     
     
         11 . The method of  claims 8 , wherein the cell is located in the central nervous system. 
     
     
         12 . The method of  claim 8 , wherein the mammalian cell is a neuronal cell, an astrocyte, or an oligodendrocyte. 
     
     
         13 . The method of  claim 8 , wherein the cell is in the spinal cord. 
     
     
         14 . The method of  claim 8 , wherein the cell is in the retina or optic nerve. 
     
     
         15 . A method of treating a CNS injury, comprising contacting the injury with an amount of the PTEN sdRNA of  claim 1  effective to promote axon regeneration. 
     
     
         16 . A method of treating a CNS injury, comprising contacting the injury with an amount of the PTEN sdRNA of  claim 2  effective to promote axon regeneration. 
     
     
         17 . A method of treating a CNS injury, comprising contacting the injury with an amount of the PTEN sdRNA of  claim 6  effective to promote axon regeneration. 
     
     
         18 . A method of treating a CNS injury, comprising contacting the injury with an amount of the PTEN sdRNA of  claim 6  effective to promote astrocyte cell migration to, and proliferation at, the injury. 
     
     
         19 . The method of  claim 15 , wherein the CNS injury is a spinal cord injury or an optic neuropathy. 
     
     
         20 . The method of  claim 15 , wherein plasticity of interneurons is promoted at the injury. 
     
     
         21 . A method for promoting the survival or regeneration of a mature CNS neuron, the neuron having an axonal injury,
 the method comprising contacting the injured neuron with a therapeutically effective amount of PTEN sdRNA.   
     
     
         22 . The method of  claim 21 , wherein the injured neuron is in the spinal cord of a mammalian subject.