Methods for selectively modulating the activity of distinct subtypes of cells

Abstract

The present invention relates to pseudotyped retrovirus-like particles or retroviral vectors comprising both engineered envelope glycoproteins derived from a virus of the Paramyxoviridae family fused to a cell targeting domain and fused to a functional domain. The present invention also relates to the use of said pseudotyped retrovirus-like particles or retroviral vectors to selectively modulate the activity of specific subsets of cells, in particular of specific immune cells. These pseudotyped retrovirus-like particles or retroviral vectors are particularly useful for gene therapy, immune therapy and/or vaccination.

1 . A method for selectively transducing target cells, wherein the method comprises contacting a pseudotyped retrovirus-like particle or retroviral vector with target cells, wherein the pseudotyped retrovirus-like particle or retroviral vector comprises:
 a) at least one cell targeting fusion protein comprising (i) a protein that is a truncated envelope glycoprotein G of a Nipah virus that lacks amino acids 2-34 of SEQ ID NO: 9 (GcΔ34) and (ii) at least one cell targeting domain; and   b) at least one truncated envelope glycoprotein F of a Nipah virus that lacks amino acids 525-546 of SEQ ID NO:11 (FcΔ22).   
     
     
         2 . The method of  claim 1 , wherein the truncated envelope glycoprotein G of a Nipah virus comprises one or more point mutations selected from the group consisting of E501A, W504A, Q530A, and E533A by comparison to the sequence of SEQ ID NO:9. 
     
     
         3 . A method for selectively transducing target cells, wherein the method comprises contacting a pseudotyped retrovirus-like particle or retroviral vector with target cells, wherein the pseudotyped retrovirus-like particle or retroviral vector comprises:
 a) at least one cell targeting fusion protein comprising (i) a protein that is a truncated envelope glycoprotein G of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein G and has at least 80% sequence identity to SEQ ID NO: 9 and (ii) at least one cell targeting domain;   b) at least one truncated envelope glycoprotein F of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein F and has at least 80% sequence identity to SEQ ID NO:11; and   (c) a nucleic acid encoding a chimeric antigen receptor.   
     
     
         4 . The method of  claim 3 , wherein the truncated envelope glycoprotein G of a Nipah virus lacks amino acids 2-34 of SEQ ID NO: 9 (GcΔ34). 
     
     
         5 . The method of  claim 4 , wherein the truncated envelope glycoprotein G of a Nipah virus comprises one or more point mutations selected from the group consisting of E501A, W504A, Q530A, and E533A by comparison to the sequence of SEQ ID NO:9. 
     
     
         6 . The method of  claim 3 , wherein the at least one envelope glycoprotein F of a Nipah virus lacks amino acids 525-546 of SEQ ID NO:11 (FcΔ22). 
     
     
         7 . The method of  claim 3 , wherein:
 the envelope glycoprotein G of a Nipah virus lacks amino acids 2-34 of SEQ ID NO: 9 (GcΔ34) and comprises the point mutations E501A, W504A, Q530A, and E533A by comparison to the sequence of SEQ ID NO:9; and   the at least one envelope glycoprotein F of a Nipah virus lacks amino acids 525-546 of SEQ ID NO:11 (FcΔ22).   
     
     
         8 . The method of  claim 1 , wherein the retrovirus-like particle or retroviral vector is a lentiviral-like particle or lentiviral vector. 
     
     
         9 . The method of  claim 1 , wherein the at least one cell targeting domain is specific for CD3, CD8, or CD4. 
     
     
         10 . The method of  claim 3 , wherein the at least one cell targeting domain is specific for CD3, CD8 or CD4. 
     
     
         11 . The method of  claim 1 , wherein the at least one cell targeting domain is a Darpin or an scFv. 
     
     
         12 . The method of  claim 1 , wherein the pseudotyped retrovirus-like particle or retroviral vector comprises a gene of interest encoding a therapeutic protein, apoptotic protein, cell surface receptor, antibody, antibody fragment, shRNA, antigen, cytokine, microRNA, CRISPR (clustered, regularly interspaced, short palindromic repeat)/CAS element(s), a Zinc Finger Nucleases, or a S/MAR (Scaffold / Matrix Attachment Region) episomes, ligand and/or receptor. 
     
     
         13 . The method of  claim 1 , which comprises a gene encoding a chimeric antigen receptor. 
     
     
         14 . The method of  claim 3 , wherein the encoded chimeric antigen receptor is specific for CD19. 
     
     
         15 . The method of  claim 1 , wherein the retrovirus-like particle or retroviral vector comprises at least one modulating fusion protein comprising (i) a protein derived from an envelope glycoprotein G or Hof a virus of the  Paramyxoviridae  family or a transmembrane domain and (ii) at least one functional domain. 
     
     
         16 . The method of  claim 15 , wherein the at least one functional domain is a cytokine, growth factor, hormone, neurotransmitter, apoptosis ligand, or a combination thereof. 
     
     
         17 . The method of  claim 15 , wherein the modulating fusion protein comprises IL-7. 
     
     
         18 . A method of performing immune therapy on a subject, comprising administering to the subject an effective amount of a pseudotyped retrovirus-like particle or retroviral vector, wherein
 the pseudotyped retrovirus-like particle or retroviral vector comprises
 a) at least one cell targeting fusion protein comprising (i) a protein that is a truncated envelope glycoprotein G of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein G and has at least 80% sequence identity to SEQ ID NO: 9 and (ii) at least one cell targeting domain; 
 b) at least one truncated envelope glycoprotein F of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein F and has at least 80% sequence identity to SEQ ID NO:11; and 
 c) a chimeric antigen receptor. 
   
     
     
         19 . A method of treating a subjecting having cancer, comprising administering to the subject an effective amount of a pseudotyped retrovirus-like particle or retroviral vector, wherein the pseudotyped retrovirus-like particle or retroviral vector comprises
 a) at least one cell targeting fusion protein comprising (i) a protein that is a truncated envelope glycoprotein G of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein G and has at least 80% sequence identity to SEQ ID NO: 9 and (ii) at least one cell targeting domain;   b) at least one truncated envelope glycoprotein F of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein F and has at least 80% sequence identity to SEQ ID NO:11; and   c) a chimeric antigen receptor.   
     
     
         20 . The method of  claim 18 , wherein the chimeric antigen receptor is specific for CD19. 
     
     
         21 . A method for producing a pseudotyped retrovirus-like particle or retroviral vector, wherein said method comprises co-transfecting a packaging cell line with:
 (i) at least one nucleic acid encoding a cell targeting fusion protein, said cell targeting fusion protein comprising (i) a truncated envelope glycoprotein G of a Nipah virus that has a truncation in the cytoplasmic region and comprises at least 80% sequence identity to SEQ ID NO: 9 and (ii) at least one cell targeting domain,   (ii) at least one nucleic acid encoding a modulating fusion protein, said modulating fusion protein comprising (i) a truncated envelope glycoprotein G of a Nipah virus that has a truncation in the cytoplasmic region and comprises at least 80% sequence identity to SEQ ID NO: 9 or a transmembrane domain (ii) at least one functional domain,   (iii) at least one nucleic acid encoding a protein that is a truncated envelope glycoprotein F of a Nipah virus that lacks at least one part of the cytoplasmic region of said envelope glycoprotein F and has at least 80% sequence identity to SEQ ID NO: 11, and   (iv) at least one vector comprising a nucleic acid encoding core proteins from said retrovirus, wherein said cell targeting fusion protein and/or said modulating fusion protein comprises a truncated envelope glycoprotein G of a Nipah virus that has a truncation in the cytoplasmic region and comprises at least 80% sequence identity to SEQ ID NO: 9.