Antibody based gene therapy with tissue-directed expression

Abstract

Embodiments of the disclosure include methods and compositions for treatment of a medical condition related to the liver, including at least viral infections and liver cancer, for example. In specific embodiments, immunotherapies are provided for delivering polynucleotides locally to the liver, wherein the polynucleotides encode particular gene products that include bispecific antibodies, including those that target certain liver antigens, for example.

What is claimed is: 
     
         1 . A method of producing an antibody against hepatitis B virus (HBV) in situ in a tissue within a subject, comprising the steps of:
 a) administering two or more polynucleotides directly to the tissue within the subject, wherein each of the two or more polynucleotides each encoding non-identical monospecific antibody polypeptides; and   b) generating antibodies in situ in the tissue within the subject, wherein the generated antibodies dimerize to each other to produce a mixture of monospecific antibody or bispecific antibody polypeptides against HBV.   
     
     
         2 . The method of  claim 1 , wherein the monospecific antibody or the bispecific antibody comprises one or more antigen-binding domains, wherein each of the antigen-bending domains comprising a single chain antibody, a single chain variable fragment (scFv), peptide, camelid variable domain, shark IgNAR variable domain, single domain antibody, affimer or VHH antibody. 
     
     
         3 . The method of  claim 1 , wherein the polynucleotides further encode at least one liver antigen-targeting entity and/or immunostimulatory entity. 
     
     
         4 . The method of  claim 3 , wherein the liver antigen-targeting entity comprises a single chain antibody, a single chain variable fragment, a camelid antibody, or a peptide. 
     
     
         5 . The method of  claim 3 , wherein the immunostimulatory entity comprises an anti-CD3 scFv, an anti-CD28 scFv, anti-41BB scFv, anti-OX40 scFv, anti-CTLA4 scFv, anti-CD16 scFv, anti-PD1 scFv, anti-PD-L1 scFv, anti-CD47 scFv, part or all of the ectodomain for a ligand for CD28, part or all of the ectodomain of 41BB ligand, part or all of the ectodomain of the LIGHT protein, ICOS-ligand, CD276 (B7-H3), B7-H4, B7-H6, CD134L, CD137L, or a cytokine. 
     
     
         6 . The method of  claim 1 , wherein the polynucleotides are included in a vector. 
     
     
         7 . The method of  claim 6 , wherein the vector is adeno-associated virus (AAV), wherein the vector comprises a tissue-specific promoter. 
     
     
         8 . The method of  claim 1 , wherein the antibody against (HBV) is specific to HBV small surface antigen, HBV middle surface antigen, HBV large surface antigen. 
     
     
         9 . A method of producing a bispecific, trispecific or quadraspecific antibodies in situ in a tissue within a subject, comprising the steps of:
 a) administering two or more polynucleotides directly to the tissue within the subject, wherein each of said two or more polynucleotides encodes non-identical bispecific antibody polypeptides, wherein the antibodies produced from the polynucleotides in situ in the tissue of the subject dimerize to each other, and   b) producing a mixture of bispecific, trispecific or quadraspecific antibodies in situ in the tissue within the subject.   
     
     
         10 . A method of treating hepatitis B in a subject having HBV infection, comprising the steps of:
 a) administering a therapeutically effective amount of a vector directly to the tissue within the subject, wherein the vector comprising two or more polynucleotides each encoding non-identical monospecific antibody polypeptides;   b) generating antibodies in situ in the tissue within the subject, wherein the non-identical monospecific antibody polypeptides dimerize to each other to produce a mixture of bispecific antibody polypeptides against HBV and having immunostimulating capacity; and   c) reducing the HBV viral load within the tissue.   
     
     
         11 . The method of  claim 10 , wherein the bispecific antibody comprises one or more antigen-binding domains, wherein each of the antigen-bending domains comprising a single chain antibody, a single chain variable fragment (scFv), peptide, camelid variable domain, shark IgNAR variable domain, single domain antibody, affimer or VHH antibody. 
     
     
         12 . The method of  claim 10 , wherein the vector encodes at least one liver antigen-targeting entity and/or immunostimulatory entity. 
     
     
         13 . The method of  claim 12 , wherein the liver antigen-targeting entity comprises a single chain antibody, a single chain variable fragment, a camelid antibody, or a peptide. 
     
     
         14 . The method of  claim 12 , wherein the immunostimulatory entity comprises an anti-CD3 scFv, an anti-CD28 scFv, anti-41BB scFv, anti-OX40 scFv, anti-CTLA4 scFv, anti-CD16 scFv, anti-PD1 scFv, anti-PD-L1 scFv, anti-CD47 scFv, part or all of the ectodomain for a ligand for CD28, part or all of the ectodomain of 41BB ligand, part or all of the ectodomain of the LIGHT protein, ICOS-ligand, CD276 (B7-H3), B7-H4, B7-H6, CD134L, CD137L, or a cytokine.