Protoparvovirus and tetraparvovirus compositions and methods for gene therapy
AssigneeSynteny Therapeutics, Inc.
InventorsRobert Kotin, Sebastian Aguirre
Life of this patent
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Abstract
Disclosed are recombinant virions that have a capsid protein, or a variant thereof, of a protoparvovirus or tetraparvovirus, and a nucleic acid that includes a heterologous nucleic acid.
Claims176
A recombinant virion comprising (1) at least one capsid protein or a variant thereof, of a protoparvovirus or a genotypic variant thereof, and (2) a nucleic acid, wherein the nucleic acid comprises a heterologous nucleic acid.
A recombinant virion comprising (1) at least one capsid protein or a variant thereof, of a tetraparvovirus or a genotypic variant thereof, and (2) a nucleic acid, wherein the nucleic acid comprises a heterologous nucleic acid.
The recombinant virion of any one of the preceding claims, wherein the virion is icosahedral.
The recombinant virion of any one of the preceding claims, wherein the capsid protein or a variant thereof comprises structural proteins VP1 and/or VP2.
The recombinant virion of any one of preceding claims, wherein the heterologous nucleic acid comprises a nucleic acid sequence that is at least about 60% identical to a nucleic acid sequence of a target cell.
The recombinant virion of any one of preceding claims, wherein the heterologous nucleic acid is at least about 60% identical to the nucleic acid of a mammal, preferably wherein the mammal is a human.
The recombinant virion of any one of preceding claims, wherein the heterologous nucleic acid is not operably linked to a protoparvovirus or tetraparvovirus promoter.
The recombinant virion of any one of preceding claims, wherein the nucleic acid comprises at least one inverted terminal repeat (ITR).
The recombinant virion of any one of the preceding claims, wherein the nucleic acid is deoxyribonucleic acid (DNA).
The recombinant virion of any one of the preceding claims, wherein the nucleic acid comprises a Rep protein-dependent origin of replication (ori).
The recombinant virion of any one of the preceding claims, wherein the nucleic acid comprises a nucleic acid operably linked to a promoter, optionally placed between two ITRs.
The recombinant virion of any one of the preceding claims, wherein the heterologous nucleic acid encodes a coding RNA and/or a non-coding RNA.
The recombinant virion of any one of the preceding claims, wherein the nucleic acid comprises a non-coding DNA.
The recombinant virion of any one of preceding claims, wherein the nucleic acid comprises a nucleic acid sequence that is at least about 80% identical to the nucleic acid sequence of a genomic safe harbor (GSH) of the target cell.
The recombinant virion of any one of the preceding claims, wherein the nucleic acid is integrated into the genome of a target cell upon transduction.
The recombinant virion of any one of the preceding claims, wherein the nucleic acid comprises a nucleic acid sequence encoding at least one replication protein and capsid protein or a variant thereof.
The recombinant virion of any one of the preceding claims, wherein the virion is autonomously replicating.
The recombinant virion of any one of the preceding claims, wherein the virion binds and/or transduces a cancer cell or non-cancerous cell.
The recombinant virion of any one of preceding claims, wherein the virion binds and/or transduces a stem cell (e.g., hematopoietic stem cell, CD34+ stem cell, CD36+ stem cell, mesenchymal stem cell, cancer stem cell).
The recombinant virion of any one of the preceding claims, wherein the recombinant virion binds and/or transduces a hematopoietic cell, hematopoietic progenitor cell, hematopoietic stem cell, erythroid lineage cell, megakaryocyte, erythroid progenitor cell (EPC), CD34+ cell, CD36+ cell, mesenchymal stem cell, nerve cell, intestinal cell, intestinal stem cell, gut epithelial cell, endothelial cell, lung cell, enterocyte, liver cell (e.g., hepatocyte, hepatic stellate cells (HSCs), Kupffer cells (KCs), liver sinusoidal endothelial cells (LSECs)), brain microvascular endothelial cell (BMVECs), erythroid progenitor cell, lymphoid progenitor cell, B lymphoblast cell, B cell, T cell, basophilic Endemic Burkitt Lymphoma (EBL), polychromatic erythroblast, epidermal stem cell, P63-positive keratinocyte-derived stem cell, keratinocyte, pancreatic β-cell, K cell, L cell, and/or orthochromatic erythroblast.
The recombinant virion of any one of the preceding claims, wherein the at least one capsid protein or a variant thereof comprises a VP2 sequence having one or more mutations with respect to canine parvovirus strain N (UniProtKB—P12930) or the amino acid sequence SEQ ID NO:
- 27.
The recombinant virion of any one of the preceding claims, wherein the at least one capsid protein or a variant thereof comprises a heterologous peptide tag.
A pharmaceutical composition comprising the recombinant virion of any one of the preceding claims;
- and a carrier and/or a diluent.
An insect cell, comprising at least one vector, comprising:
- (i) a nucleotide sequence comprising at least one ITR nucleotide sequence,
- (ii) a nucleotide sequence comprising at least one gene encoding the protoparvovirus or tetraparvovirus capsid proteins VP1, VP2, and/or a variant thereof, operably linked to at least one expression control sequence for expression in an insect cell, and
- (iii) a nucleotide sequence comprising
- (A) at least one replication protein of a protoparvovirus or tetraparvovirus operably linked to at least one expression control sequence for expression in an insect cell,
- (B) at least one replication protein of an AAV, optionally wherein the at least one replication protein of an AAV comprises (a) a Rep52 or a Rep40 coding sequence operably linked to at least one expression control sequence for expression in an insect cell, and/or (b) a Rep78 or a Rep68 coding sequence operably linked to at least one expression control sequence for expression in an insect cell, or
- (C) a combination of (A) and (B).
Dependencies are inferred from the claim text.