Bioactive FUS1 peptides and nanoparticle-polypeptide complexes
View Patent ↗A nanoparticle-polypeptide complex comprising a bioactive polypeptide in association with a nanoparticle, wherein the bioactive polypeptide is modified by the addition of a chemical moiety that facilitates cellular uptake of the protein. The polypeptide can be a protein or a peptide. In some embodiments, the amino acid sequence of the protein or peptide is derived from the amino acid sequence of a tumor suppressor gene product.
1. A method of delivering a FUS-1 polypeptide to a cell, comprising contacting a cell with a nanoparticle-polypeptide complex comprising one or more FUS-1 polypeptides, wherein the one or more FUS-1 polypeptides:
(1) comprise the sequence of KLRRVHKNLPPQGPVKLDHPR (SEQ ID NO:13);
(2) comprise the sequence of VLRRVHVNLPPQGPVVLDHPR (SEQ ID NO:14);
(3) comprise the sequence of ETPVTVNGQVRAPLRRVHV (SEQ ID NO:17);
(4) consists of the sequence of KLRRVHKNLIPQGIVKLDHPR (SEQ ID NO:1); or
(5) consists of the sequence of ETIVTKNGQKRAKLRRVHK (SEQ ID NO:16)
wherein the one or more FUS-1 polypeptides are each modified by the addition of a chemical moiety to facilitate uptake into the cell.
2. The method of claim 1 , wherein the FUS-1 polypeptide contains no more than 30 amino acids.
3. The method of claim 2 , wherein the FUS-1 polypeptide contains no more than 24 amino acids.
4. The method of claim 1 , wherein the FUS-1 polypeptide comprises KLRRVHKNLPPQGPVKLDHPR (SEQ ID NO:13).
5. The method of claim 1 , wherein the FUS-1 polypeptide comprises VLRRVHVNLPPQGPVVLDHPR (SEQ ID NO:14).
6. The method of claim 1 , wherein the FUS-1 polypeptide comprises ETPVTVNGQVRAPLRRVHV (SEQ ID NO:17).
7. The method of claim 1 , wherein the FUS-1 polypeptide consists of KLRRVHKNLIPQGIVKLDHPR (SEQ ID NO:1).
8. The method of claim 1 , wherein the FUS-1 polypeptide consists of ETIVTKNGQKRAKLRRVHK (SEQ ID NO:16).
9. The method of claim 1 , wherein the nanoparticle is a lipid-based nanoparticle, a superparamagnetic nanoparticle, a nanoshell, a semiconductor nanocrystal, a quantum dot, a polymer-based nanoparticle, a silicon-based nanoparticle, a silica-based nanoparticle, a metal-based nanoparticle, a fullerene or a nanotube.
10. The method of claim 9 , wherein the nanoparticle is a lipid-based nanoparticle.
11. The method of claim 9 , wherein the lipid-based nanoparticle is a liposome, a neutral liposome, a DOPC liposome or a DOTAP:cholesterol vesicle.
12. The method of claim 11 , wherein the liposome is a DOPC liposome.
13. The method of claim 9 , wherein the nanoparticle is a superparamagnetic nanoparticle.
14. The method of claim 9 , wherein the nanoparticle is a superparamagnetic nanoparticle, and the nanoparticle-polypeptide complex is within a liposome or a DOTAP:cholesterol vesicle.
15. The method of claim 14 , wherein the liposome is a DOPC liposome.
16. The method of claim 1 , wherein the chemical moiety is a C 4 -C 18 fatty acid.
17. The method of claim 16 , wherein the fatty acid is stearate or myristate.
18. The method of claim 1 , wherein each FUS-1 polypeptide is coupled or fused to a cell-penetrating peptide selected from the group consisting of an HIV Tat peptide, a herpes virus VP22 peptide, and a Drosophila Antennapedia homeobox gene product.
19. The method of claim 1 , wherein the FUS-1 polypeptide is attached to a C 4 -C 18 fatty acid that is a stearate moiety, and the nanoparticle is a DOTAP:cholesterol vesicle, a DOPC liposome or a superparamagnetic nanoparticle.
20. The method of claim 1 , wherein the FUS-1 polypeptide is a peptide of from 19 to 100 amino acid residues in length.
21. The method of claim 20 , wherein the peptide is from 21 to 100 amino acid residues in length.
22. The method of claim 1 , wherein the cell is in a subject, and contacting comprises administering to the subject a composition comprising the nanoparticle-polypeptide complex.
23. The method of claim 1 , wherein the subject has a tumor, and the cell is a tumor cell.
24. The method of claim 23 , wherein the tumor is a lung cancer.
25. The method of claim 24 , wherein the lung cancer is a NSCLC.
26. The method of claim 22 , wherein administering comprises intraperitoneal injection.
27. The method of claim 23 , wherein the tumor is gastrointestinal.
28. The method of claim 1 , wherein the chemical moiety to facilitate uptake into the cell is a fatty acid.
29. The method of claim 1 , wherein the chemical moiety to facilitate uptake into the cell is a cell penetrating peptide.
30. The method of claim 29 , wherein the cell penetrating peptide is an HIV Tat peptide, a herpes virus VP22 peptide, or a Drosophila Antennapedia homeobox gene product.
31. A method of delivering a FUS-1 polypeptide to a cell, comprising contacting a cell with a nanoparticle-polypeptide complex comprising a FUS-1 polypeptide consisting of KLRRVHKNLIPQGIVKLDHPR (SEQ ID NO:1), wherein the FUS-1 polypeptide is modified at the N-terminal Lysine residue by either (i) the attachment of a C 4 -C 18 fatty acid to facilitate cellular uptake or (ii) the coupling or fusing of a cell-penetrating peptide selected from the group consisting of an HIV Tat peptide, a herpes virus VP22 peptide, and a Drosophila Antennapedia homeobox gene product.