Compositions of a peptide targeting system for treating cancer
This invention describes a protein nanoparticle system for targeting siRNA or other drugs into tumors. The basis of the protein system is elastin-like peptides that self-assemble once exposed to the nucleic acid of the siRNA. Specific targeting peptides are fused to the core ELP structure by standard genetic engineering techniques. These targeting peptides confer specific binding of the nanoparticle to receptors on the surface of tumor cells and allow for uptake of the nanoparticle into the tumor cells.
1. An isolated polypeptide comprising an assembly domain (AD), a cell targeting domain (CTD), and a nucleic acid binding domain (NBD), wherein the isolated polypeptide comprises SEQ ID No. 56.
2. The polypeptide of claim 1 , further comprising a drug binding domain (DBD).
3. The polypeptide of claim 1 , wherein the nucleic acid binding domain (NBD) binds to a siRNA molecule.
4. The peptide of claim 1 , wherein the NBD comprises a cationic domain.
5. The polypeptide of claim 2 , wherein the DBD binds a conventional or investigational drug.
6. An isolated polypeptide comprising an assembly domain (AD), a cell targeting domain (CTD), and a nucleic acid binding domain (NBD), wherein the CTD comprises SEQ ID No. 19.
7. An isolated cDNA polynucleotide comprising a nucleotide sequence that encodes the polypeptide of claim 1 , wherein the isolated polynucleotide comprises SEQ ID No. 55.
8. A recombinant expression construct comprising the polynucleotide of claim 7 .
9. An isolated host cell comprising the polynucleotide of claim 7 .
10. A nanoparticle comprising one or more polypeptides according to claim 1 , wherein the nanoparticle assembles in solution.
11. The nanoparticle of claim 10 , further comprising a drug agent.
12. The nanoparticle of claim 11 , wherein the drug agent is a siRNA, shRNA, miRNA, antisense nucleic acid, single-stranded nucleic acid, double-stranded nucleic acid, DNA expression vector, RNA expression vector, plasmid, viral vector, oligonucleotide, or chemical compound.
13. A method of targeting a drug agent to a target cell, comprising:
obtaining one or more polypeptides comprising an assembly domain (AD), a nucleotide binding domain (NBD), and a cell targeting domain (CTD);
contacting the drug agent with the NBD of one or more polypeptides;
assembling the AD of one or more polypeptides to generate an assembled polypeptide;
contacting the assembled polypeptide with the target cell; and
contacting the CTD of the assembled polypeptide with a biological molecule present outside of the target cell,
wherein one or more of the polypeptides comprises SEQ ID No. 56.
14. The method of claim 13 , wherein the drug agent comprises a siRNA, shRNA, miRNA, antisense nucleic acid, single-stranded nucleic acid, double-stranded nucleic acid, DNA expression vector, RNA expression vector, plasmid, viral vector, oligonucleotide, or chemical compound.
15. The method of claim 13 , wherein the cell comprises a tumor cell.
16. The method of claim 15 , wherein the tumor cell is a human bladder cell, a human breast cell, a human colon cell, a human liver cell, a human lung cell, a human neuroblastoma cell, a human ovarian cell, a human pancreatic cell, a human prostate cell, or a human skin cell.
17. The nanoparticle of claim 10 , wherein the nanoparticle assembles in solution when combined with nucleic acid.
18. An isolated cDNA polynucleotide encoding the polypeptide of claim 1 .
19. An isolated cDNA polynucleotide encoding the polypeptide of claim 6 .