Collagen-mimetic peptide mediated delivery of nucleic acid carriers for efficient delivery from collagen
The present invention provides a composition for delivering a polynucleotide into cells via a polyplex. The polyplex comprises the polynucleotide and a polymer. The composition comprises the polyplex, a collagen-mimetic peptide (CMP) and collagen fragments. The CMP is bound to the polyplex and the collagen fragments. Also provided are the uses of the composition in methods of delivering a polynucleotide into cells as well as methods of improving wound healing in a subject, enhancing cell proliferation in a subject, enhancing production of extracellular matrix by cells in a subject and/or enhancing cell migration by cells in a subject.
1. A method comprising:
(a) administering an effective amount of a composition to cells in a subject, wherein the composition comprises
(i) at least one collagen-mimetic peptide (CMP),
(ii) a polyplex, wherein the polyplex comprises at least one polymer and at least one polynucleotide encoding a protein, and the at least one CMP is bound to the polyplex, wherein the protein is a healing factor selected from the group consisting of a growth factor, an anti-inflammatory cytokine, and a combination thereof, and
(iii) collagen or fragments thereof, wherein the at least one CMP is bound to the collagen or fragments thereof;
(b) expressing the protein by the cells in the subject; and
(c) enhancing migration, proliferation and/or differentiation of the cells in the subject.
2. The method of claim 1 , wherein the at least one polynucleotide comprises DNA.
3. The method of claim 1 , wherein the at least one polynucleotide comprises RNA.
4. The method of claim 1 , wherein the at least one CMP is bound to the polyplex via a covalent linkage.
5. The method of claim 1 , wherein the at least one CMP is bound to the polyplex via a noncovalent linkage.
6. The method of claim 1 , wherein the at least one polynucleotide further encodes at least one silencing RNA capable of suppressing expression of the protein, the method further comprising suppressing the expression of the protein by the cells.
7. The method of claim 1 , wherein the cells are at a wound in the subject.
8. The method of claim 1 , wherein step (c) comprises enhancing proliferation of the cells.
9. The method of claim 1 , further comprising enhancing production of extracellular matrix by the cells.
10. The method of claim 1 , wherein step (c) comprises enhancing migration of the cells.
11. The method of claim 7 , wherein the wound is a chronic wound.
12. The method of claim 1 , wherein the protein is a growth factor.
13. The method of claim 1 , wherein the protein is an anti-inflammatory cytokine.
14. The method of claim 1 , wherein the composition further comprises an extracellular matrix (ECM) component selected from the group consisting of fibrin, fibronectin, laminins, ECM proteoglycans, ECM glycosaminoglycans and combinations thereof.
15. A method comprising:
(a) administering an effective amount of a composition to cells in a subject, wherein the composition comprises
(i) at least one collagen-mimetic peptide (CMP),
(ii) a polyplex, wherein the polyplex comprises at least one polymer and at least one polynucleotide encoding a protein, and the at least one CMP is bound to the polyplex,
(iii) collagen or fragments thereof, wherein the at least one CMP is bound to the collagen or fragments thereof, and
(iv) a matrix metalloproteinase (MMP); and
(b) expressing the protein by the cells in the subject.
16. The method of claim 1 , wherein the at least one CMP is (GPP)3GPRGEKGERGPR(GPP)3GPCCG (SEQ ID NO: 3) or (GPO)4GEKGER(GPO)4GGCG (SEQ ID NO: 4).
17. The method of claim 1 , wherein step (c) comprises enhancing differentiation of the cells in the subject.