Replication-defective arenavirus vectors
The invention relates to an infectious arenavirus particle that is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells. One or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells, and foreign genes coding for an antigen or other protein of interest or nucleic acids modulating host gene expression are expressed under control of the arenavirus promoters, internal ribosome entry sites or under control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III. The modified arenaviruses are useful as vaccines and therapeutic agents for a variety of diseases.
1. An infectious arenavirus particle engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells, wherein the arenavirus open reading frame glycoprotein (GP) is removed and replaced by a foreign ribonucleic acid sequence selected from short hairpin RNAs (shRNA), small interfering RNA (siRNA) and micro RNAs (miRNA).
2. An infectious arenavirus particle, wherein the arenavirus particle is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells, wherein the arenavirus particle comprises a foreign ribonucleic acid sequence coding for an antigen selected from the group consisting of respiratory syncytial virus antigens, human immunodeficiency virus antigens, hepatitis C virus antigens, varizella zoster virus antigens, herpes simplex virus antigens, cytomegalovirus antigens and antigens derived from mycobacterium tuberculosis.
3. A pharmaceutical preparation comprising an arenavirus particle according to claim 1 or 2 .
4. The arenavirus particle according to claim 1 or 2 , wherein an internal ribosome entry sites is introduced into the viral genome.
5. The arenavirus particle according to claim 1 or 2 , wherein the arenavirus is lymphocytic choriomeningitis virus (LCMV).
6. A method of treating infections caused by viruses, bacteria, parasites and prions, autoimmune diseases, neoplastic diseases, metabolic diseases, degenerative diseases, inherited diseases, allergic diseases, or substance dependence in a patient comprising administering a therapeutically effective amount of an arenavirus particle according to claim 1 or 2 to a patient in need thereof.
7. A method of expressing a protein of interest or modifying gene expression in a cell culture wherein the cell culture is infected with an arenavirus particle according to claim 1 or 2 .
8. The arenavirus particle of claim 2 , wherein the GP gene of the arenavirus is engineered such that the arenavirus is unable to produce further infectious progeny particles in normal, not genetically engineered cells, and wherein the open reading frames for the NP protein and the RNA-dependent RNA polymerase L of the arenavirus are intact.
9. The arenavirus particle of claim 2 , wherein the open reading frame of the GP gene of the arenavirus has been removed.
10. The areanavirus particle of claim 2 comprising a modified genome, wherein
i) one or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells; and
ii) the foreign ribonucleic acid sequence is expressed under control of one or more of the four arenavirus promoters 5′ UTR, and 3′ UTR of the S segment, and 5′ UTR and 3′ UTR of the L segment, and/or under control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III, expressed either by themselves or as read-through by fusion to arenavirus protein open reading frames.