Antigenic compositions and use of same in the targeted delivery of nucleic acids
View Patent ↗Methods and compositions are provided for delivery of therapeutic nucleic acids to a target cell. A chimeric antigen is provided to encapsulate, bind, or otherwise carry a nucleic acid molecule to a target cell where the chimeric antigen and nucleic acid are internalized by receptor-mediated endocytosis. The chimeric antigen has a nucleic acid interaction domain, a target binding domain, and an immune response domain that may include a target antigen. Targeting is generally provided by the specificity of the target binding domain for a particular target cell receptor, but may also be provided by inclusion of a targeting antigen within the immune response domain. The combined delivery of chimeric antigen and nucleic acid, which may be a siRNA, may be synergistic in certain applications, for example in breaking host tolerance to a virus or in providing immunostimulation.
1. A method for inhibiting expression of a target gene within a target cell, the method comprising the steps of:
providing a nucleic acid molecule suitable for effecting RNAi of a target gene;
providing fusion protein comprising
a target binding domain for binding to a receptor on the surface of the target cell, the target binding domain comprising a hinge region, at least a portion of a C H 1 region and an Fc antibody fragment comprising a C H 2 and a C H 3 domain;
an immune response domain operatively attached to the target binding domain;
a first nucleic acid interaction domain corresponding to a HBV core protein or a fragment thereof operatively attached to the immune response domain; and
a second nucleic acid interaction domain operatively attached at the C-terminus of the target binding domain corresponding to the HBV core protein or a fragment thereof having at least the protamine domain of the HBV core protein; and
administering the nucleic acid molecule and the fusion protein to contact the target cell.
2. The method as in claim 1 , wherein the nucleic acid molecule is mixed with a suitable amount of the fusion protein to create a nucleic acid delivery complex prior to contacting the target cell.
3. The method as in claim 1 , wherein the HBV core protein fragment is the assembly domain of HBV core protein.
4. The method as in claim 1 , wherein the nucleic acid binding domain is operatively attached to the C-terminus of the target binding domain.
5. The method as in claim 1 , wherein the target cell is a mammalian host cell, and wherein the step of administering the nucleic acid and fusion protein to the target cell comprises administering said nucleic acid and fusion protein to the mammalian host.
6. The method as in claim 5 , wherein the target binding domain comprises a xenotypic antibody fragment.
7. The method of claim 1 , wherein the first nucleic acid binding domain comprises a fragment of the assembly domain of the HBV core protein and wherein the second nucleic acid binding domain comprises a fragment of the assembly domain and the protamine domain of the HBV core protein.
8. The method of claim 1 , wherein the first nucleic acid binding domain comprises at least amino acids 1 to 78 of SEQ ID NO:2 of the HBV core protein and wherein the second nucleic acid binding domain comprises at least amino acids 81 to 183 of SEQ ID NO:2 of the HBV core protein.
9. The method of claim 1 , wherein the immune response domain provides targeting to a secondary target cell.