Targeting tumor microenvironments with recombinant prokaryotic collagen-like proteins
A recombinant collagen-like protein comprising a binding domain having binding capacity for both extra domain A and extra domain B-containing variants of cellular fibronectin. Cancer may be treated BY administering the recombinant collagen-like protein to a patient.
1. A recombinant collagen-like protein comprising a binding domain having binding capacity for both extra domain A and extra domain B-containing variants of cellular fibronectin,
wherein the protein comprises:
a recombinant collagen-like (CL)-domain, and
a variable (V)-domain comprising an ECM-binding domain from a streptococcal Scl1.1 protein;
wherein the recombinant CL-domain is a CL-domain from a streptococcal Scl2.28 protein;
wherein the CL-domain from the streptococcal Scl2.28 protein is modified to include an integrin recognition sequence from the CL-domain of the streptococcal Scl1.41 protein.
2. The recombinant collagen-like protein of claim 1 , wherein:
the protein is a streptococcal collagen-like protein from a Group A Streptococcus , and
the Group A Streptococcus is Streptococcus pyogenes.
3. The recombinant collagen-like protein of claim 1 , wherein the protein further comprises binding capacity for Tenascin-C.
4. The recombinant collagen-like protein of claim 1 , wherein the protein recognizes collagen-binding integrin receptors α 2 β 1 and α 11 β 1 .
5. The recombinant collagen-like protein of claim 1 , wherein the protein comprises two copies of an integrin recognition sequence GLPGER.
6. The recombinant collagen-like protein of claim 5 , wherein the protein comprises the variable domain of the rScl1.1 sequence of SEQ ID NO: 3.
7. A recombinant collagen-like protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
8. A pharmaceutical composition comprising
the recombinant collagen-like protein of claim 7 ; and
a pharmaceutically acceptable carrier.
9. The composition of claim 8 , wherein the carrier is a nanoparticle.
10. The composition of claim 9 , wherein the nanoparticle is coated with the collagen-like protein.
11. A recombinant collagen-like protein comprising a binding domain having binding capacity for both extra domain A and extra domain B-containing variants of cellular fibronectin,
wherein the protein comprises:
a recombinant collagen-like (CL)-domain, and
a variable (V)-domain;
wherein:
the recombinant CL-domain is a CL-domain from a streptococcal Scl2.28 protein, and
the CL-domain from the streptococcal Scl2.28 protein is modified to include two integrin recognition sequences from the CL-domain of the streptococcal Scl1.41 protein.
12. The recombinant collagen-like protein of claim 11 , wherein the two integrin recognition sequences each comprise SEQ ID NO: 6.
13. The recombinant collagen-like protein of claim 11 , wherein:
the variable (V)-domain comprises an ECM-binding domain from a streptococcal Scl1.1 protein; or
the variable (V)-domain is a variable domain from a streptococcal Scl1.1 protein.
14. A method of targeting a cancer in a patient in need thereof, comprising administering a recombinant collagen-like protein of claim 7 to the patient.
15. A method of treating cancer in a patient in need thereof, comprising administering a recombinant collagen-like protein of claim 1 to the patient, wherein the collagen-like protein is conjugated with a therapeutic agent useful for the treatment of cancer.
16. A method of targeting a cancer in a patient in need thereof, comprising administering a recombinant collagen-like protein of claim 7 to the patient, wherein the cancer is selected from a group consisting of a tumor or a metastatic cancer.
17. A method of treating cancer in a patient in need thereof, comprising administering a recombinant collagen-like protein of claim 7 to the patient, wherein the cancer is selected from a group consisting of breast cancer, prostate cancer, melanoma, gastric cancer, colorectal cancer, and head and neck cancer, wherein the collagen-like protein is conjugated with a therapeutic agent useful for the treatment of cancer.
18. A method of preparing a collagen-like protein comprising a binding domain having the capacity to bind to both extra domain A and extra domain B-containing variants of cellular fibronectin comprising the steps of combining:
(i) two copies of an integrin recognition sequence of SEQ ID NO: 6 inserted into the collagen-like (CL) domain of a protein of SEQ ID NO: 4; and
(ii) the variable (V)-domain from a protein of SEQ ID NO: 3.