Modified paramyxoviridae fusion glycoproteins
AssigneeSana Biotechnology, Inc.
InventorsChristopher BANDORO, Lauren Pepper MACKENZIE, Jagesh Vijaykumar SHAH, Kyle Marvin TRUDEAU
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Abstract
Provided herein are lipid particles, such as lentiviral particles, that incorporate or are pseudotyped with a variant Nipah Virus F (NiV-F) envelope glycoprotein, and in some aspects also an attachment glycoprotein (G) protein such as a NiV-G protein or a biologically active portion or variant thereof. Also provided are polynucleotides encoding the variant NiV-F and producer cells for preparation of the lipid particles, such as lentiviral particles, containing the variant NiV-F proteins, as well as methods for preparing and using the lipid particles, such as lentiviral particles.
Claims275
A lipid particle, comprising:
- (a) a lipid bilayer;
- (b) a paramyxovirus glycoprotein (G protein) or a biologically active portion thereof;
- and
- (c) a variant Nipah virus F glycoprotein (NiV-F) comprising a modified cytoplasmic tail, wherein the modified cytoplasmic tail comprises:
- (i) a heterologous cytoplasmic tail or a truncated portion thereof from a glycoprotein from another virus or a virus-associated protein, wherein the variant NiV-F is a chimeric protein;
- (ii) a truncated NiV-F cytoplasmic tail that has a deletion of between 23 and 27 contiguous amino acid residues at or near the C-terminus of the wild-type NiV-F protein cytoplasmic tail set forth in SEQ ID NO:4, with the proviso that the truncated NiV-F does not have a deletion of 25 contiguous amino acids;
- and/or
- (iii) a modified NiV-F cytoplasmic that comprises a modified endocytosis motif,
- wherein the G protein or the biologically active portion thereof and the variant NiV-F protein are exposed on the outside of the lipid bilayer.
A pseudotyped lentiviral particle, comprising:
- (a) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (b) a variant Nipah virus F glycoprotein (NiV-F) comprising a modified cytoplasmic tail, wherein the modified cytoplasmic tail comprises:
- (i) a heterologous cytoplasmic tail or a truncated portion thereof from a glycoprotein from another virus or a virus-associated protein, wherein the variant NiV-F is a chimeric protein;
- (ii) a truncated NiV-F cytoplasmic tail that has a deletion of between 23 and 27 contiguous amino acid residues at or near the C-terminus of the wild-type NiV-F protein cytoplasmic tail set forth in SEQ ID NO:4, with the proviso that the truncated NiV-F does not have a deletion of 25 contiguous amino acids;
- and/or
- (iii) a modified NiV-F cytoplasmic that comprises a modified endocytosis motif,
- wherein the G protein or the biologically active portion thereof and the variant NiV-F protein are exposed on the outside of the lipid bilayer.
A lipid particle, comprising:
- (a) a lipid bilayer;
- (b) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (c) a variant Nipah virus F glycoprotein (NiV-F) comprising a modified ectodomain, wherein the modified ectodomain comprises a modified cleavage site selected from:
- (i) replacement of the cleavage sequence NNTHDLVGDVRLAGV (SEQ ID NO:311) with a modified furin cleavage sequence;
- or
- (ii) replacement of the cleavage sequence VGDVR (SEQ ID NO:339) with a modified cleavage sequence from another paramyxovirus, optionally wherein the modified cleavage sequence from another paramyxovirus comprises a cathepsin L cleavage site.
A pseudotyped lentiviral particle, comprising:
- (a) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (b) a variant Nipah virus F glycoprotein (NiV-F) comprising a modified ectodomain, wherein the modified ectodomain comprises a modified cleavage site selected from:
- (i) replacement of the cleavage sequence NNTHDLVGDVRLAGV (SEQ ID NO:311) with a modified furin cleavage sequence;
- or;
- or
- (ii) replacement of the cleavage sequence VGDVR (SEQ ID NO:329) VGDVR (SEQ ID NO:339) with a modified cleavage sequence from another paramyxovirus, optionally wherein the cleavage sequence is a cathepsin L cleavage site.
A lipid particle, comprising:
- (a) a lipid bilayer;
- (b) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (c) a variant Nipah virus F glycoprotein (NiV-F) comprising a heterologous signal sequence, or encoded by a polynucleotide sequence comprising a sequence that encodes a heterologous signal sequence, compared to the wild-type signal sequence of NiV-F.
A pseudotyped lentiviral particle, comprising:
- (a) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (b) a variant Nipah virus F glycoprotein (NiV-F) comprising a heterologous signal sequence, or encoded by a polynucleotide sequence comprising a sequence that encodes a heterologous signal sequence, compared to the wild-type signal sequence of NiV-F.
A lipid particle, comprising:
- (a) a lipid bilayer;
- (b) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (c) a variant Nipah virus F glycoprotein (NiV-F) comprising a heterologous or modified transmembrane domain compared to wild-type NiV-F.
A pseudotyped lentiviral particle, comprising:
- (a) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (b) a variant Nipah virus F glycoprotein (NiV-F) comprising a heterologous or modified transmembrane domain compared to wild-type NiV-F.
A lipid particle, comprising:
- (a) a lipid bilayer;
- (b) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (c) a variant Nipah virus F glycoprotein (NiV-F) comprising a hyperfusogenic mutation selected from:
- a mutation that is introduced at a N-glycosylation site with respect to amino acid positions 64, 67, 99, 414 and/or 464 of SEQ ID NO. 1;
- and/or
- a mutation that is introduced at a hexamer stability site with respect to amino acid positions 393 and/or 109 of SEQ ID NO. 1.
A pseudotyped lentiviral particle, comprising:
- (a) a paramyxovirus glycoprotein (G) or a biologically active portion thereof;
- and
- (b) a variant Nipah virus F glycoprotein (NiV-F) comprising a hyperfusogenic mutation selected from:
- a mutation that is introduced at a N-glycosylation site with respect to amino acid positions 64, 67, 99, 414, and/or 464 of SEQ ID NO. 1;
- and/or
- a mutation that is introduced at a hexamer stability site with respect to amino acid positions 393 and/or 109 of SEQ ID NO. 1.
A polynucleotide comprising a nucleic acid molecule encoding a variant Nipah virus F glycoprotein (NiV-F) comprising a modified cytoplasmic tail, wherein the modified cytoplasmic tail comprises:
- (i) a heterologous cytoplasmic tail or a truncated portion thereof from a glycoprotein from another virus or a virus-associated protein, wherein the variant NiV-F is a chimeric protein;
- (ii) a truncated NiV-F cytoplasmic tail that has a deletion of between 23 and 27 contiguous amino acid residues at or near the C-terminus of the wild-type NiV-F protein cytoplasmic tail set forth in SEQ ID NO:4, with the proviso that the truncated NiV-F does not have a deletion of 25 contiguous amino acids;
- and/or
- (iii) a modified NiV-F cytoplasmic that comprises a modified endocytosis motif.
A polynucleotide comprising a nucleic acid molecule encoding a variant Nipah virus F glycoprotein (NiV-F) comprising a modified ectodomain, wherein the modified ectodomain comprises a modified cleavage site selected from:
- (i) replacement of the cleavage sequence NNTHDLVGDVRLAGV (SEQ ID NO:311) with a modified furin cleavage sequence;
- or
- (ii) replacement of the cleavage sequence VGDVR (SEQ ID NO:339) with a modified cleavage sequence from another paramyxovirus, optionally wherein the cleavage sequence is a cathepsin L cleavage site;
- wherein the replacement is in the sequence of amino acids set forth in SEQ ID NO:
- 1 or SEQ ID NO:320, or a mature form thereof lacking the signal peptide (amino acids 1-26).
A polynucleotide comprising a nucleic acid molecule encoding a variant Nipah virus F glycoprotein (NiV-F) comprising a heterologous signal sequence compared to the wild-type signal sequence of NiV-F.
A polynucleotide comprising a nucleic acid molecule encoding a variant Nipah virus F glycoprotein (NiV-F) comprising a heterologous or modified transmembrane domain compared to wild-type NiV-F.
A polynucleotide comprising a nucleic acid molecule encoding a variant Nipah virus F glycoprotein (NiV-F) comprising a hyperfusogenic mutation selected from:
- a mutation that is introduced at a N-glycosylation site with respect to amino acid positions 64, 67, and/or 99 of SEQ ID NO. 1;
- and/or
- a mutation that is introduced at a hexamer stability site with respect to amino acid positions 393 and/or 109 of SEQ ID NO. 1.
Dependencies are inferred from the claim text.