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 method for treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a non-covalent complex of (a) an antibody or antigen-binding fragment thereof, and (b) a therapeutic polynucleotide,
wherein the antibody or antigen-binding fragment thereof comprises 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 V L CDRs having the amino acid sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26;
and wherein the cancer comprises one or more cells that express a functional ENT2 transporter.
2. The method 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 method according to claim 2 , wherein the V H has the amino acid sequence of SEQ ID NO:2 and the V L has the amino acid sequence of SEQ ID NO:7 or 8.
4. The method 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 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 method according to claim 4 , wherein the V H has an amino acid sequence 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 method according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a humanized antibody.
7. The method according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a monovalent, divalent, or multivalent single chain variable fragment (scFv).
8. The method according to claim 1 , wherein the antibody or antigen-binding fragment thereof is a bispecific antibody.
9. The method according to 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 method according to claim 1 , wherein the therapeutic polynucleotide is a polynucleotide inhibitor of protein translation.
11. The method according to claim 10 , wherein the polynucleotide inhibitor of protein translation is an antisense oligonucleotide.
12. The method according to claim 10 , wherein the polynucleotide inhibitor of protein translation is selected from the group consisting of an siRNA, an miRNA, a shRNA, and an external guide sequence (EGS).
13. The method according to claim 10 , wherein the polynucleotide inhibitor targets mRNA encoding an oncogenic protein.
14. The method according to claim 10 , wherein the composition comprises:
a first complex of (a) an antibody or antigen-binding fragment thereof comprising V H CDRs having the amino acid sequences of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18, and 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 first polynucleotide inhibitor of protein translation; and
a second complex of (a) an antibody or antigen-binding fragment thereof comprising V H CDRs having the amino acid sequences of SEQ ID NO:16, SEQ ID NO:17, and SEQ ID NO:18, and 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 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.
15. The method according to claim 1 , wherein the therapeutic polynucleotide is an mRNA or plasmid.
16. The method according to claim 15 , wherein the mRNA encodes a polypeptide ligand for a receptor of an immune cell.
17. The method according to claim 16 , wherein the polypeptide ligand stimulates the immune system of the subject.
18. The method according to claim 15 , wherein the mRNA encodes an antigen.
19. The method according to claim 18 , wherein the antigen is a viral antigen.
20. The method according to claim 1 , wherein the cancer is selected from the group consisting of a pancreatic cancer, a skin cancer, a brain cancer, a colorectal cancer, and a breast cancer.
21. The method according to claim 1 , wherein the cancer is selected from the group consisting of a vascular cancer, multiple myeloma, an adenocarcinoma, a sarcoma, a bone cancer, a bladder cancer, a cervical cancer, an esophageal cancer, a kidney cancer, a liver cancer, a lung cancer, a nasopharangeal cancer, a prostate cancer, a stomach cancer, and a uterine cancer.
22. The method according to claim 1 , wherein the composition further comprises a pharmaceutically acceptable excipient.