Invasion complex and methods of targeting

• US 20030044863A1
• Filed: 07/18/2002
• Published: 03/06/2003
• Est. Priority Date: 07/20/2001
• Status: Abandoned Application

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Ashkar, Samy
• Application Number:
  US10/197,725
• Publication Number:
  US 20030044863A1
• Time in Patent Office: Days
• US Class Current: 435/7.23
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C40B 40/02 Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors

Abstract

Therapeutics have been identified and developed with are targeted to inhibiting metastasis. These are based on the discovery of an invasion complex which confers the ability of cells, for example tumor cells, to translocate across extracellular matrix barriers, as well as the identification of novel peptides that interact with the invasion complex and regulate it'"'"'s activity. Whole or partial complexes, or individual molecules of the invasion complex are used to screen for proteins or compounds that interact with the invasion complex. Methods of screening for interacting proteins such as osteopontin, sophin B (SEQ ID NO: 1), or compounds are well known in the art, some of which are described below. Once interacting proteins have been identified, they are screened for inhibition, enhancement, or reduction of complex activity. The presence of proteins of the invasion complex on or within cells is indicative of particular diseases or disorders, for example, those characterized by a tumor cell or activated macrophage. Elevated levels of proteins that interact with the invasion complex, such as osteopontin, are also indicative of cancer, and in particular, metastatic cancer. Diagnostic methods, described below, will assist in the identification of subjects having, or at risk of developing a disorder associated with aberrant expression of any of the identified proteins that form the invasion complex. Because this invasion complex confers the ability to translocate across matrix barriers, peptides that bind to the complex play a role in regulating metastasis and/or organ specific homing of cancer cells.

Claims

1. A method for identifying cancer in a patient comprising

(a) isolating cells or tissue from the patient;

and

(b) identifying an invasion complex, or one or more invasion complex components, associated with the cells or tissue.

2. The method of claim 1 wherein the invasion complex is associated with or identified by an assay for cell migration.

3. The method of claim 1 wherein the invasion complex is associated with or identified by an assay for tumorogenicity.

4. The method of claim 1 wherein the invasion complex comprises a homing component, a proteolytic enzyme and interacts with a ligand, such as osteopontin.

5. The method of claim 1 wherein the invasion complex is obtained by immunoreaction with an antibody to a protein selected from the group consisting of integrin B-1, P 13 kinase, SHP-1, integrin associated kinase, focal adhesion kinase, caveolin, and CD44.

6. The method of claim 5 wherein the invasion complex is obtained by immunoreaction with an antibody to a component selected from the group consisting of integrin β

-1, integrin β

-3, CD44, and integrin β

-5.

7. The method of claim 1 wherein the invasion complex components are over-expressed, underexpressed, or mutated in the cells or tissue as compared to normal cells or tissue.

8. The method of claim 1 wherein the invasion complex components are identified as nucleic acid sequences.

9. The method of claim 8 wherein the nucleic acid sequences encode a protein selected from the group consisting of CD44(v3-v6), Integrin B1, P13 Kinase, SHP-1, Integrin Associated Kinase, focal adhesion kinase (FAK), Caveolin, tissue plasminogen activator, urokinase plasminogen activator, matrix metalloproteinase 2, matrix metalloproteinase 9, mbMMP (membrane bound metalloproteinase), PKC , PKC 6, paxillin, rhoB, and G₁I.

10. The method of claim 1 wherein the invasion complex components are proteins selected from the group consisting of CD44(v3-v6), Integrin B 1, P13 Kinase, SHP-1, Integrin Associated Kinase, focal adhesion kinase (FAK), Caveolin, tissue plasminogen activator, urokinase plasminogen activator, matrix metalloproteinase 2, matrix metalloproteinase 9, mbMMP (membrane bound metalloproteinase), PKC β

, PKC δ

, paxillin, rhoB, and G₁I.

13. The invasion complex of claim 1 wherein the invasion complex is associated with or identified by an assay for tumorogenicity.

11. An isolated invasion complex, or portion thereof comprising more than one protein.

12. The invasion complex claim 11 wherein the invasion complex is associated with or identified by an assay for cell migration.

14. The invasion complex of claim 11 wherein the invasion complex comprises a homing component, a proteolytic enzyme and interacts with a ligand, such as osteopontin.

15. The invasion complex of claim 11 wherein the invasion complex is obtained by immunoreaction with an antibody to a protein selected from the group consisting of integrin B-1, P13 kinase, SHP-1, integrin associated kinase, focal adhesion kinase, caveolin, and CD44.

16. The invasion complex of claim 15 wherein the invasion complex is obtained by immunoreaction with an antibody to integrin B-1.

17. The invasion complex of claim 11 wherein the invasion complex components are over-expressed, underexpressed, or mutated in the cells or tissue as compared to normal cells or tissue.

18. The invasion complex of claim 11 wherein the invasion complex components are identified as nucleic acid sequences.

19. The invasion complex of claim 18 wherein the nucleic acid sequences encode a protein selected from the group consisting of CD44(v3-v6), Integrin B1, PI3 Kinase, SHP-1, Integrin Associated Kinase, focal adhesion kinase (FAK), Caveolin, tissue plasminogen activator, urokinase plasminogen activator, matrix metalloproteinase 2, matrix metalloproteinase 9, mbMMP (membrane bound metalloproteinase), PKC β

, PKC δ

, paxillin, rhoB, and G₁l.

20. The invasion complex of claim 11 wherein the invasion complex components are proteins selected from the group consisting of CD44(v3-v6), Integrin B 1, PI3 Kinase, SHP-1, Integrin Associated Kinase, focal adhesion kinase (FAK), Caveolin, tissue plasminogen activator, urokinase plasminogen activator, matrix metalloproteinase 2, matrix metalloproteinase 9, mbMMP (membrane bound metalloproteinase), PKC β

, PKC δ

, paxillin, rhoB, and G₁I.

21. A method for inhibiting cancer metastasis comprising

providing a compound that inhibits the assembly or an activity of an invasion complex as defined by claim 11, or a component thereof.

22. A method for identifying compounds useful in the diagnosis or inhibition of cancer metastasis, comprising

identifying compounds that interact with an invasion complex or one or more of the proteins of the invasion complex.

23. The method of claim 22 further comprising determining if the compounds alter the assembly or an activity of the invasion complex.

24. The method of claim 22 further comprising determining the levels of compound present in an individual or sample comprising the invasion complex.

25. The method of claim 22 further comprising:

(a) isolating cells or tissue from the individual; and

(b) identifying one or more invasion complex components in the cells or tissue.

26. The method of claim 25 wherein the invasion complex components are identified as nucleic acid sequences.

27. The method of claim 26 wherein the nucleic acid sequences encode a protein selected from the group consisting of CD44(v3-v6), Integrin B 1, PI3 Kinase, SHP-1, Integrin Associated Kinase, focal adhesion kinase (FAK), Caveolin, tissue plasminogen activator, urokinase plasminogen activator, matrix metalloproteinase 2, matrix metalloproteinase 9, mbMMP (membrane bound metalloproteinase), PKC β

, PKC δ

, paxillin, rhoB, and G₁I.

28. The method of claim 25 wherein the invasion complex components are proteins selected from the group consisting of CD44(v3-v6), Integrin B1, PI3 Kinase, SHP-1, Integrin Associated Kinase, focal adhesion kinase (FAK), Caveolin, tissue plasminogen activator, urokinase plasminogen activator, matrix metalloproteinase 2, matrix metalloproteinase 9, mbMMP (membrane bound metalloproteinase), PKC β

, PKC δ

, paxillin, rhoB, and G₁I.

29. An isolated compound which interfers with the assembly or an activity of the invasion complex or a component thereof.

30. A diagnostic agent selectively reactive with an invasion complex or an isolated component thereof.

31. The agent of claim 30 wherein the agent is labeled for detection in an assay.