Compositions and methods for delivery of nucleic acids to cells
Compositions and methods of use thereof for delivering nucleic acid cargo into cells are provided. The compositions typically include (a) a 3E10 monoclonal antibody or an antigen binding, cell-penetrating fragment thereof; a monovalent, divalent, or multivalent single chain variable fragment (scFv); or a diabody; or humanized form or variant thereof, and (b) a nucleic acid cargo including, for example, a nucleic acid encoding a polypeptide, a functional nucleic acid, a nucleic acid encoding a functional nucleic acid, or a combination thereof. Elements (a) and (b) are typically non-covalently linked to form a complex.
1. A composition comprising a non-covalent complex of
(a) an antibody or antigen-binding fragment thereof comprising heavy chain variable region (V H ) complementarity determining regions (CDRs) having the amino acid sequences of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18, and light chain variable region (V L ) CDRs having the amino acid sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26, and
(b) a polynucleotide inhibitor of protein translation.
2. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (V H ) having an amino acid sequence that is at least 95% identical to SEQ ID NO:2 and a light chain variable region (V L ) having an amino acid sequence that is at least 95% identical to SEQ ID NO:7 or 8.
3. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a V H having the amino acid sequence of SEQ ID NO:2 and a V L having the amino acid sequence of SEQ ID NO:7 or 8.
4. The composition according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a V H having an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 6 and a V L having an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 and 53-55.
5. The composition according to claim 4 , wherein the V H has an amino acid sequence selected from the group consisting of SEQ ID NO: 6 and the V L has an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 and 53-55.
6. The composition of claim 1 , wherein the antibody or antigen-binding fragment thereof is a humanized antibody.
7. The composition of claim 1 , wherein the antibody or antigen-binding fragment thereof is a monovalent, divalent, or multivalent single chain variable fragment (scFv).
8. The composition of claim 1 , wherein the antibody or antigen-binding fragment thereof is a bispecific antibody.
9. The composition of claim 8 , wherein the bispecific antibody comprises:
a first heavy chain or antigen-binding fragment thereof comprising CDRs having the amino acid sequences of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18, and a first light chain or antigen binding fragment thereof comprising CDRs having the amino acid sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26; and
a second heavy chain, or antigen-binding fragment thereof, and a second light chain, or antigen binding fragment thereof, that associate to specifically bind a target cell-type, tissue, or organ.
10. The composition according to claim 1 , wherein the polynucleotide inhibitor of protein translation is an antisense oligonucleotide.
11. The composition according to claim 1 , wherein the polynucleotide inhibitor of protein translation is an siRNA.
12. The composition according to claim 1 , wherein the polynucleotide inhibitor of protein translation is an miRNA.
13. The composition according to claim 1 , wherein the polynucleotide inhibitor of protein translation is an shRNA.
14. The composition according to claim 1 , wherein the polynucleotide inhibitor of protein translation is an external guide sequence (EGS).
15. The composition according to claim 1 , wherein the polynucleotide inhibitor targets mRNA encoding an oncogenic protein.
16. The composition according to claim 1 , wherein the composition comprises:
a first complex of (a) the antibody or antigen-binding fragment thereof, and (b) a first polynucleotide inhibitor of protein translation; and
a second complex of (a) the antibody or antigen-binding fragment thereof, and (b) a second polynucleotide inhibitor of protein translation;
wherein the first polynucleotide inhibitor of protein translation and the second inhibitor of protein translation are different inhibitors of protein translation.
17. The composition according to claim 1 , further comprising a pharmaceutically acceptable excipient.