Method for treating a disorder associated with Eya dysregulation by modulating Eya phosphatase activity
The present invention provides a novel mechanistic pathway and methods related to this pathway for the identification of compounds for the treatment of diseases involving cell proliferation, invasion and/or metastasis such as cancer. In particular, the instant invention relates to the phosphatase activity of Eya and the Eya-Six complex as a target for identifying novel therapeutic agents for the treatment of proliferative, invasive and/or metastatic disorders, and compositions identified using the methods disclosed herein.
1. A method for identifying a potential therapeutic agent for the treatment of a disorder associated with Eyes Absent dysregulation, comprising the steps of:
i) providing an Eya protein or fragment thereof, wherein the Eya protein or fragment possesses relevant biochemical activity, and a substrate capable of being dephosphorylated by the Eya protein or fragment;
ii) delivering a test molecule to the Eya protein or fragment, wherein the test molecule is a small molecule, peptide, polypeptide, or antibody;
iii) determining the effect of the test molecule on the phosphatase activity of the Eya protein or fragment; and
iv) comparing the phosphatase activity of the Eya protein or fragment in the presence of a test molecule to that of a reference standard;
wherein a test molecule that increases or decreases Eya phosphatase activity is a potential therapeutic agent for the treatment of a disorder associated with Eya dysregulation.
2. The method of claim 1 , further comprising the steps of:
i) providing a Six protein or fragment thereof, wherein the Six protein or fragment possesses relevant biochemical activity; and
ii) delivering a test molecule to the Six protein or fragment, wherein the test molecule is a small molecule, peptide, polypeptide, or antibody.
3. The method according to claim 1 wherein the disorder associated with Eya dysregulation is a proliferative disorder.
4. The method according to claim 1 wherein the disorder associated with Eya dysregulation is an invasive and/or metastatic disorder.
5. The method according to claim 1 wherein the disorder associated with Eya dysregulation is a angiogenic disorder.
6. The method according to claim 1 wherein the disorder associated with Eya dysregulation is a vascular disorder.
7. The method according to claim 1 wherein the disorder associated with Eya dysregulation is cancer.
8. The method according to claim 1 wherein the Eya protein is selected from the group consisting of Eya1, Eya2, Eya3 and Eya4.
9. The method according to claim 1 wherein the fragment includes the ED region of an Eya protein selected from the group consisting of Eya1, Eya2, Eya3, and Eya4.
10. The method according to claim 2 wherein the Six protein is selected from the group consisting of Six1, Six2, Six3, Six4, Six5, and Six6.
11. The method according to claim 1 wherein the substrate is selected from the group consisting of the model substrate pNPP, phospho-amino acids, phosphorylated peptides or phosphorylated proteins.