Fibronectin: growth factor chimeras
Isolated protein complexes are provided comprising growth factors such as IGF-I, IGF-II, EGF, bFGF, or KGF and fibronectin, or at least domains thereof that enable binding to and activation of both a growth factor receptor, and an integrin receptor-binding domain of fibronectin. These protein complexes include synthetic proteins where the growth factor and fibronectin sequences are joined by a linker sequence. Also provided are uses of these protein complexes for stimulating or inducing cell migration and/or proliferation in wound healing, tissue engineering, cosmetic and therapeutic treatments such as skin replacement, skin replenishment and treatment of burns where epithelial cell migration is required. In other embodiments, the invention provides inhibition of cancer cell metastasis, particularly in relation to breast cancer.
1. An isolated protein complex in the form of a synthetic chimeric protein consisting of an amino acid sequence of:
(i) a growth factor or at least a domain of said growth factor which is capable of binding a cognate growth factor receptor;
(ii) an optional linker sequence; and
(iii) domain 10 of fibronectin (FN), wherein domain 10 consists of amino acids 1447-1536 of the amino acid sequence set forth in SEQ ID NO: 1.
2. The isolated protein complex of claim 1 , wherein said growth factor is selected from insulin-like growth factor- 1 (IGF-I), insulin-like growth factor-II (IGF-II), epidermal growth factor (ESE), and basic fibroblast growth factor (bFGF).
3. The isolated protein complex of claim 1 , wherein the linker sequence comprises a protease cleavage site.
4. The isolated protein complex of claim 1 , wherein the linker sequence is selected from the group consisting of:
(SEQ ID NO: 7)
(i)
Gly 4 Ser
(SEQ ID NO: 8)
(ii)
Gly 4 Ser 3
(SEQ ID NO: 9)
(iii)
(Gly 4 Ser) 3
(SEQ ID NO: 7)
(iv)
(Gly 4 Ser) 4
(SEQ ID NO: 11)
(iv)
Leu Ile Lys Met Lys Pro;
and
(SEQ ID NO: 12)
(vi)
Gln Pro Gln Gly Leu Ala Lys.
5. An isolated nucleic acid encoding the isolated protein complex of claim 1 .
6. A genetic construct, comprising the isolated nucleic acid of claim 5 operably linked to one or more regulatory nucleotide sequences in a vector.
7. The genetic construct of claim 6 , which is an expression construct, wherein the isolated nucleic acid is operably linked to a promoter.
8. An isolated host cell, comprising the genetic construct of claim 6 .
9. A pharmaceutical composition, comprising the isolated protein complex of claim 1 and a pharmaceutically-acceptable carrier, diluent or excipient.
10. A surgical implant, scaffold or prosthesis impregnated, coated or otherwise comprising the isolated protein complex of claim 1 .
11. A wound or burn dressing, comprising the isolated protein complex of claim 1 .