Assays and methods for determining the responsiveness of an individual subject to a therapeutic agent
Provided herein are methods for determining the functional status of a cellular pathway in a diseased cell sample obtained from an individual subject. These methods involve contacting a diseased cell sample obtained from the subject with a perturbing agent (e.g., an activating agent) known to perturb a specific cellular pathway when the pathway is functioning normally. A change in one or more physiological response parameters in the presence of the perturbing agent indicates that the cellular pathway targeted by the perturbing agent is functional in the individual subject. Methods of selecting a targeted therapeutic agent for an individual subject are also provided.
1. A method of treating a subject, the method comprising:
administering to the subject a targeted therapeutic agent that targets a cellular signaling pathway, wherein the cellular signaling pathway has been selected by a method comprising:
contacting a diseased cell sample comprising live whole cells obtained from the subject with a receptor ligand known to agonize the cellular signaling pathway when the pathway is functioning normally;
measuring cell adhesion or attachment in the live whole cells in the sample; and
determining by analysis of the measurements whether a change in cell adhesion or attachment occurs in the diseased cell sample in the presence of the receptor ligand, relative to a control comprising a sample of the diseased cells in the absence of the receptor ligand;
wherein a change in cell adhesion or attachment in the presence of the receptor ligand, relative to the control, indicates that the cellular signaling pathway is active in the cells obtained from the subject and that the subject is suitable to receive a targeted therapeutic agent that targets the cellular signaling pathway,
wherein the targeted therapeutic agent is selected from the group consisting of: lapatinib, afatinib, erlotinib, neratinib, crizotinib, trametinib, GSK-1120212, GSK-1059615, dasatinib, imatinib, nilotinib, tivozanib, axitinib, sunitinib, pazopanib, sorafenib, regorafenib, vemurafenib, dabrafenib, palbociclib, LDK378, trastuzumab, pertuzumab, cetuximab, panitumumab, necitumumab, sym004, rituximab, ofatumumab, ibritumomab, obinutuzumab, tositumomab, brentuximab vedoton, daratumumab, alemtuzumab, bevacizumab, ramucirumab, and combinations thereof.
2. The method of claim 1 , wherein the measurements of cell adhesion or attachment in the live whole cells in the sample is done on a continuous basis and the subsequent analysis is done on the continuous measurements.
3. The method of claim 1 , wherein the diseased cell sample is a cancer cell sample.
4. The method of claim 3 , wherein the cancer is selected from the group consisting of breast cancer, lung cancer, and colon cancer.
5. The method of claim 1 , wherein the change in cell adhesion or attachment is assessed using non-linear Euclidean analysis.
6. The method of claim 5 , wherein the Euclidean analysis is selected from the group consisting of arithmetic summation of the difference at multiple time points, temporal maxima, temporal minima, time to reach maxima or minima, changes in slope, absolute drop in biosensor signal, a total of all measurements, and combinations thereof.
7. The method of claim 5 , wherein the change in cell adhesion or attachment is measured by a change in temporal maxima or minima.