Methods for determining signal transduction activity in tumors
AssigneesYale University, HistoRx, Inc.
InventorsMark Gustavson, Jennifer Giltnane, Marisa P. Dolled-Filhart, Robert L. Camp, David L. Rimm
Life of this patent
Estimating the term…
Abstract
The method of the invention pertains to determining the signal transduction activity in a tissue section by immunohistochemistry techniques. The expression level of the receptor of interest is determined as well as the expression levels of one or more effector molecules of the receptor signal transduction pathway. Furthermore a combined ratio of expression levels of effector molecules in subcellular compartments with the receptor expression was found to have prognostic significance.
Claims32
A method for measuring activity of a signal transduction pathway in a tissue section comprising:
- a) staining the tissue section with a reagent to detect a cell receptor protein;
- and
- b) staining the tissue section with a second reagent to detect an effector molecule, the effector molecule being a member of the signal transduction pathway related to the cell receptor protein;
- c) using an optical microscope, obtaining a high resolution digital image of the stained cell receptor protein in the tissue section and of the stained effector molecule in the tissue section;
- and
- d) analyzing the digital image to quantitate an amount of the cell receptor protein present in the tissue section and an amount of effector molecule present in the tissue section;
- thereby determining the signal transduction activity in the tissue section.
An immunohistochemical method for measuring activity of a signal transduction pathway in a tissue section comprising:
- a) staining the tissue section with a reagent to detect a cell receptor protein;
- and
- b) staining the tissue section with a second reagent to detect an effector molecule, the effector molecule being a member of the signal transduction pathway related to the cell receptor protein;
- c) staining the tissue section with a third reagent to label a subcellular compartment;
- d) using a microscope, obtaining a high resolution digital image of:
- i) the stained cell receptor protein in the tissue section;
- ii) the stained effector molecule in the tissue section;
- and
- iii) the labeled subcellular compartment;
- e) analyzing the digital image to determine a qualitative amount of:
- i) the cell receptor protein present in the tissue section;
- ii) the effector molecule present in the tissue section;
- and
- iii) the effector molecule present in a subcellular compartment;
- f) analyzing the quantitative amount of each of the cell receptor, the effector molecule and the effector molecule in the subcellular compartment in relationship to one another;
- thereby determining the signal transduction activity in the tissue section.
An immunohistochemical method for measuring activity of a signal transduction pathway in a tissue section comprising:
- a) staining the tissue section with a reagent to detect a cell receptor protein;
- and
- b) staining the tissue section with one or more additional reagents to detect one or more of a plurality of effector molecules, each of the effector molecules being a member of the signal transduction pathway related to the cell receptor protein;
- c) staining the tissue section with a label to define a subcellular compartment;
- d) using an optical microscope, obtaining a high resolution digital image or a plurality of high resolution images of:
- i) the stained cell receptor protein in the tissue section;
- ii) one or more of the stained effector molecule in the tissue section;
- and
- iii) the labeled subcellular compartment;
- e) analyzing the digital image to determine a qualitative amount of:
- i) the cell receptor protein present in the tissue section;
- ii) one or more of the effector molecules present in the tissue section;
- and
- iii) one or more of the effector molecule present in the subcellular compartment;
- f) analyzing the quantitative amount of each of the cell receptor, one or more of the effector molecules and one or more of the effector molecules in the subcellular compartment in relationship to one another;
- thereby determining the signal transduction activity in the tissue section.
A method for determining an EGFR signaling profile of a tissue section, comprising the steps of:
- (a) staining the tissue section with:
- (i) a reagent to detect EGFR;
- (ii) a second reagent to detect an EGFR effector molecule;
- and
- (iii) a third reagent to detect a subcellular compartment;
- (b) using a microscope, obtaining a high resolution digital image of the:
- (i) EGFR;
- (ii) EGFR effector molecule;
- and
- (iii) subcellular compartment;
- and
- (c) analyzing the digital image to quantitate an amount of:
- (i) EGFR;
- and
- (ii) EGFR effector molecule present in the subcellular compartment;
- (iii) EGFR effector molecule present outside the subcellular compartment;
- (d) determining an effector ratio of the amount determined in step c(iii) to the amount determined in step c(ii);
- (e) comparing the EGFR amount to an expression level of a control specimen;
- (f) comparing the ratio of step (d) to a ratio of the same in a control specimen.
A method for measuring activity of a signal transduction pathway in a tissue section comprising:
- a) obtaining a high resolution digital image of a tissue section stained with a reagent to detect a cell receptor protein;
- and
- b) obtaining a high resolution digital image of a tissue section stained with a second reagent to detect an effector molecule, the effector molecule being a member of the signal transduction pathway related to the cell receptor protein;
- and
- c) analyzing the digital images to quantitate an amount of the cell receptor protein present in the tissue section and an amount of effector molecule present in the tissue section;
- thereby determining the signal transduction activity in the tissue section.
Dependencies are inferred from the claim text.