Compositions and methods for improving viral vector efficiency

Abstract

The present invention relates to peptides and compositions for use in improving transduction efficiency of viruses into target cells.

1 . A peptide comprising:
 (i) a CPP moiety; and   (ii) a Beclin-derived peptide moiety having a length of between 14 and 22 amino acid residues;   
       with the proviso that said peptide is not the peptide shown in SEQ ID NO:75, SEQ ID NO:77 and SEQ ID NO:98-101. 
     
     
         2 . The peptide according to  claim 1 , wherein the CPP moiety is:
 a peptide derived from the trans-activator of transcription (tat) peptide of the Human Immunodeficiency Virus (HIV) type-1; or   a MAP peptide.   
     
     
         3 . The peptide according to  claim 1 , wherein the CPP moiety is selected in the group consisting of SEQ ID NO:1 to SEQ ID NO:62, in particular from SEQ ID NO:1 to SEQ ID NO:5, more particularly wherein the CPP moiety is selected from SEQ ID NO:1 and SEQ ID NO:4. 
     
     
         4 . The peptide according to  claim 1 , wherein the Beclin-derived peptide moiety is a Beclin-1-derived peptide moiety, such as a sequence of Beclin-1 or a functional derivative of said sequence, that includes the β1 sheet region of Beclin-1, or a Beclin-2-derived moiety such as a Beclin-2-derived moiety comprising the amino acid sequence of SEQ ID NO:89 or a functional variant thereof. 
     
     
         5 . The peptide according to  claim 4 , wherein the Beclin-1-derived peptide moiety comprises the amino acid sequence shown in SEQ ID NO:65 or a functional variant thereof. 
     
     
         6 . The peptide according to  claim 4 , wherein the Beclin-1-derived peptide moiety comprises a sequence selected in the group consisting of SEQ ID NO:65 to 73. 
     
     
         7 . The peptide according to  claim 1 , wherein the CPP moiety and the Beclin-derived peptide moiety are linked together via a linker, such as a peptide linker, more particularly a dipeptide linker such as a GG dipeptide linker. 
     
     
         8 . The peptide according to  claim 1 , wherein the peptide comprises, or consists of, the sequence shown in any one of SEQ ID NO:74, SEQ ID 76, SEQ ID NO:78 to 83 and SEQ ID NO:90 to 91. 
     
     
         9 . A method for gene therapy comprising:
 administering to a patient in need thereof a viral vector encoding a therapeutic gene and a peptide comprising
 a cell-penetrating peptide (CPP) moiety; and 
 a Beclin-derived peptide moiety. 
   
     
     
         10 . The method according to  claim 9 , wherein the viral vector is a retrovirus, in particular a lentivirus. 
     
     
         11 . The method according to  claim 10 , wherein the lentivirus is a pseudotyped lentivirus. 
     
     
         12 . The method according to  claim 9 , wherein the viral vector is a parvovirus, in particular an adeno-associated virus (AAV). 
     
     
         13 . The method according to  claim 9 , further comprising administering another viral transduction enhancer, such as vectofusin, human fibronectin fragments, various semen-derived enhancers of viral infection or peptides derived from HIV-1 envelope glycoproteins. 
     
     
         14 . The method according to  claim 9 , wherein the peptide comprises
 (i) a CPP moiety; and   (ii) a Beclin-derived peptide moiety having a length of between 14 and 22 amino acid residues, or wherein the peptide comprises or consists of the sequence shown in SEQ ID NO:75, SEQ ID NO:77 and SEQ ID NO:98-101.   
     
     
         15 . The method according to  claim 9 , wherein the peptide is used at a concentration comprised between 0.01 and 20 μM, such as between 2.5 and 20 μM. 
     
     
         16 . A complex or mixture of a virus or viral vector with a peptide comprising:
 a cell-penetrating peptide (CPP) moiety; and   a Beclin-derived peptide moiety.   
     
     
         17 . The complex or the mixture according to  claim 16 , wherein the viral vector is a retrovirus, in particular a lentivirus. 
     
     
         18 . The complex or the mixture according to  claim 17 , wherein the lentivirus is a pseudotyped lentivirus. 
     
     
         19 . The complex or the mixture according to  claim 16 , wherein the viral vector is a parvovirus, in particular an adeno-associated virus (AAV). 
     
     
         20 . The complex or the mixture according to  claim 16 , wherein the peptide comprises
 (i) a CPP moiety; and   (ii) a Beclin-derived peptide moiety having a length of between 14 and 22 amino acid residues, or wherein the peptide comprises or consists of the sequence shown in SEQ ID NO:75, SEQ ID NO:77 and SEQ ID NO:98-101.   
     
     
         21 . A nucleic acid construct comprising a polynucleotide encoding the peptide of  claim 1 . 
     
     
         22 . The nucleic acid construct of  claim 21 , further comprising one or more control sequence(s). 
     
     
         23 . A kit comprising the peptide according to  claim 1  or comprising a peptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:75 and SEQ ID NO:77, and a viral vector. 
     
     
         24 . The kit of  claim 23 , wherein the viral vector is a retrovirus, in particular a lentivirus. 
     
     
         25 . The kit of  claim 23 , wherein the lentivirus is a pseudotyped lentivirus. 
     
     
         26 . The kit of  claim 23 , wherein the viral vector is a parvovirus, in particular an adeno-associated virus (AAV). 
     
     
         27 . An in vitro method for increasing the sensitivity of a cell-based assay for detecting the presence or absence of a virus in a sample, comprising contacting the sample, or an extract from the sample, with a cell and a peptide, wherein said peptide comprises:
 a cell-penetrating peptide (CPP) moiety; and   a Beclin-derived peptide moiety;   the method further optionally comprising determining the level of a virus within the cell and/or in the cell culture medium.   
     
     
         28 . The method according to  claim 27 , wherein the peptide comprises
 (i) a CPP moiety; and   (ii) a Beclin-derived peptide moiety having a length of between 14 and 22 amino acid residues, or wherein the peptide comprises or consists of the sequence shown in SEQ ID NO:75 and SEQ ID NO:77.