Composition for delivery of genetic material

Abstract

The present invention relates to exosomes, loaded with genetic material and methods of producing them and to the use of such exosomes for delivering genetic material in vivo, in particular the use of such exosomes in methods of gene therapy or gene silencing.

1 . A method of making a pharmaceutical composition, comprising loading exosomes with a therapeutic oligonucleotide having a chemically modified backbone by transfection of the exosome with a transfection reagent, thereby making the pharmaceutical composition. 
     
     
         2 . The method according to  claim 1 , wherein the therapeutic oligonucleotide having a chemically modified backbone is one or more of the following: a transplicing oligonucleotide, a morpholino (PMO), an antisense oligonucleotide (ASO), a peptide nucleic acid (PNA), an miRNA, an artificial plasmid, an shRNA or an siRNA. 
     
     
         3 . The method according to  claim 1 , wherein the chemical backbone modification of the oligonucleotide is selected from any one or more of the following: phosphoramidate, methylphosphonate, phosphorothioate, morpholino (PMO) or peptide nucleic acid (PNA). 
     
     
         4 . The method according to  claim 1 , wherein the pharmaceutical composition further comprises a modification of the oligonucleotide selected from any one or more of thionucleotide, 2′ O methyl or 2′ methoxy-ethyl. 
     
     
         5 . The method according to  claim 1 , wherein the transfection reagent is a cationic transfection reagent. 
     
     
         6 . The method according to  claim 1 , wherein the oligonucleotide is single-stranded or double stranded. 
     
     
         7 . A method of delivering a therapeutic oligonucleotide having a chemically modified backbone in vivo comprising delivering the pharmaceutical composition to a selected tissue or cell type, wherein the pharmaceutical composition comprises an exosome that includes exogenous genetic material transfected into the exosome by a transfection reagent, wherein the exogenous genetic material is the therapeutic oligonucleotide having the chemically modified backbone. 
     
     
         8 . A method of treatment by gene silencing or gene therapy comprising administering to the patient in need thereof, an effective amount of the pharmaceutical composition, wherein the pharmaceutical composition comprises an exosome that includes exogenous genetic material transfected into the exosome by a transfection reagent, wherein the exogenous genetic material is a therapeutic oligonucleotide having a chemically modified backbone. 
     
     
         9 . The method of treatment according to  claim 8 , wherein the disease to be treated is selected from: cancer, neurodegenerative disease, muscular disorders, haemophilia, cystic fibrosis or stroke. 
     
     
         10 . The method of treatment according to  claim 9 , wherein the neurodegenerative disease to be treated is selected from: Alzheimer's disease, Huntington's disease or Parkinson's disease. 
     
     
         11 . The method of treatment according to  claim 9 , wherein the muscular disorder is selected from: Duchenne Muscular Dystrophy, spinal muscular atrophy or myotonic dystrophy.