Methods for the detection and quantitation of pten

A method for determining if PTEN expression levels are elevated or reduced in one or more tumor cells in a biological sample, said method comprising:

• a. obtaining a biological sample comprising one or more tumor cells and one or more stromal cells;
• b. assaying the biological sample for expression of PTEN by using a detectably labeled binding molecule specific for PTEN;
• c. quantitating an amount of PTEN expression in the one or more tumor cells and an amount of PTEN expression in the one or more stromal cells by determining an optical density (OD) reading for PTEN expression in the tumor cells and an OD reading for PTEN expression in the normal cells, wherein the OD reading for the tumor cells and stromal cells is quantitated from an average OD reading determined for the tumor cells and the normal cells on a per cell basis in a defined cellular area and wherein the average OD reading on a per cell basis is obtained by dividing the average OD reading for the defined cellular area by a number of nuclei in the defined cellular area;
• d. comparing the PTEN OD reading in the tumor cells to the PTEN OD reading in the stromal cells;
• and
• e. determining that PTEN is elevated in the tumor cells where the PTEN OD reading is greater in the tumor cells as compared to the stromal cells or determining that PTEN is reduced in the tumor cells where the PTEN OD reading is less in the tumor cells than in the stromal cells.

A method for predicting responsiveness of a subject with a disease or disorder to a receptor tyrosine kinase inhibitor, said method comprising:

• a. obtaining the biological sample comprising one or more tumor cells and one or more normal cells from the subject;
• b. assaying the biological sample for expression of PTEN;
• c. quantitating an amount of PTEN expression in the one or more tumor cells and an amount of PTEN expression in the one or more normal cells;
• d. comparing the amount of PTEN expression in the tumor cells to the amount of PTEN expression in the normal cells;
• e. determining that PTEN is elevated in the tumor cells where the amount of expression of PTEN is greater in the tumor cells as compared to the normal cells or determining that PTEN is reduced in the tumor cells where the amount of expression of PTEN is less in the tumor cells than in the normal cells;
• and
• f. predicting that the subject is responsive to a receptor tyrosine kinase inhibitor where expression of PTEN in the tumor cells is less than expression of PTEN in the normal cells or determining that the subject is not responsive to a receptor tyrosine kinase inhibitor where expression of PTEN in the tumor cells is the same or greater than expression of PTEN in the normal cells.

A method for treating a subject with a disease or disorder with a receptor tyrosine kinase inhibitor, the method comprising:

• a. obtaining the biological sample comprising one or more tumor cells and one or more normal cells from the subject;
• b. assaying the biological sample for expression of PTEN;
• c. quantitating an amount of PTEN expression in the one or more tumor cells and an amount of PTEN expression in the one or more normal cells;
• d. comparing the amount of PTEN expression in the tumor cells to the amount of PTEN expression in the normal cells;
• e. determining that PTEN is elevated in the tumor cells where the amount of expression of PTEN is greater in the tumor cells as compared to the normal cells or determining that PTEN is reduced in the tumor cells where the amount of expression of PTEN is less in the tumor cells than in the normal cells;
• and
• f. administering the receptor tyrosine kinase inhibitor to the subject.

A method for selecting subjects for inclusion/exclusion in a clinical trial, said method comprising:

• a. obtaining a biological sample from each subject comprising one or more tumor cells and one or more stromal cells;
• b. assaying the biological sample obtained from each subject for expression of PTEN by using a detectably labeled binding molecule specific for PTEN;
• c. quantitating an amount of PTEN expression in the one or more tumor cells and an amount of PTEN expression in the one or more stromal cells;
• d. comparing the amount of PTEN expression in the tumor cells to the amount of PTEN expression in the stromal cells;
• e. determining that PTEN expression levels are elevated in the tumor cells where the amount of PTEN expression is greater in the tumor cells as compared to the stromal cells or determining that PTEN expression is reduced in the tumor cells where the amount of PTEN expression is less in the tumor cells than in the stromal cells;
• and
• f. selecting subjects for inclusion/exclusion in the clinical trial that have an amount of PTEN expression that is greater in the tumor cells as compared to the stromal cells or selecting subjects for inclusion/exclusion in the clinical trial that have an amount of PTEN expression that is less in the tumor cells as compared to the stromal cells.