Compositions and methods of use of oncolytic virus like vesicles

A high-titer hybrid virus vector for generating oncolytic virus-like vesicles (VLVs) comprising the following operably linked sequence elements:

• a) a first DNA sequence comprising a DNA promoter sequence;
• b) a second DNA sequence encoding alphavirus non-structural protein polynucleotide sequences;
• c) a third DNA sequence encoding an alphavirus subgenomic RNA promoter;
• d) a fourth DNA sequence comprising:
• (i) a sequence domain encoding an IL-12 polypeptide,
• (ii) a sequence domain encoding an IL-17RA-DN polypeptide, and
• (iii) a sequence domain encoding a PD-L1-short hairpin RNA (shRNA),
• wherein each of the sequence domains (i)-(iii) are optionally preceded by a sequence independently selected from the group consisting of one or more subgenomic promoters, a sequence encoding a 2A peptide sequence, or combinations thereof;
• and
• e) a fifth DNA sequence encoding a vesiculovirus glycoprotein;
• wherein the titer of the high-titer hybrid virus vector is at least about 1×107 plaque forming units (pfu) per mL of virus like vesicles (VLVs), or is at least about 5×107 plaque forming units (pfu) per mL of virus like vesicles (VLVs), or is at least about 1×108 plaque forming units (pfu) per mL of virus like vesicles (VLVs).

A high-titer hybrid virus vector for treatment, prophylaxis, or prevention of malignancy or infectious disease comprising the following operably linked sequence elements:

• a) a first DNA sequence comprising a DNA promoter sequence;
• b) a second DNA sequence encoding alphavirus non-structural protein polynucleotide sequences;
• c) a third DNA sequence encoding an alphavirus subgenomic RNA promoter;
• d) a fourth DNA sequence comprising:
• (i) a sequence domain comprising SEQ ID NO. 6,
• (ii) a sequence domain comprising SEQ ID NO. 8, and
• (iii) a sequence domain comprising SEQ ID NO. 10,
• wherein each of the sequence domains (i)-(iii) are optionally preceded by a sequence independently selected from the group consisting of one or more subgenomic promoters, a sequence encoding a 2A peptide sequence, or combinations thereof;
• and
• e) a fifth DNA sequence encoding a vesiculovirus glycoprotein;
• wherein the titer of the high-titer hybrid virus vector is at least about 1×107 plaque forming units (pfu) per mL of virus like vesicles (VLVs), or is at least about 5×107 plaque forming units (pfu) per mL of virus like vesicles (VLVs), or is at least about 1×108 plaque forming units (pfu) per mL of virus like vesicles (VLVs).