Whole cell assays and methods
The disclosure provides methods for analysis of disease cell response to a therapeutic agent. In embodiments, a method comprises administering the therapeutic agent to a disease cell sample from the subject in a device that measures at least one physiological parameter of a cell; determining whether a change occurs in the physiologic parameter of the disease cell sample in response to the therapeutic agent as compared to a baseline measurement or the physiological parameter before administration of the therapeutic agent, and selecting the therapeutic agent that results in the change in the at least one physiologic parameter. In embodiments, the disease cells are whole, viable, and/or label free.
1. A method of treating a human subject diagnosed with cancer, the method comprising:
administering to the subject a first agent that is a targeted therapeutic that has been determined to be therapeutically active in the signaling pathway it is intended to address in the subject's cancer cells by a method comprising:
culturing a sample consisting essentially of viable primary or metastatic cancer cells obtained from the subject in a media free of serum;
contacting the sample with the first agent and with a second agent that is known to selectively affect the same signaling pathway the first agent is intended to address, so as to upregulate or downregulate the signaling pathway as measured by an effect on cell adhesion or attachment, to produce a sample contacted with both the first agent and the second agent;
continuously measuring cell adhesion or attachment of viable primary or metastatic cancer cells in the sample contacted with both the first agent and the second agent, relative to a sample of viable primary or metastatic cancer cells obtained from the subject which sample is contacted with the first agent or the second agent alone;
determining by mathematical analysis of the continuous measurements an output value, expressed as a percentage, that characterizes whether a change in cell adhesion or attachment has occurred in the sample contacted with both the first agent and the second agent, as compared to the sample contacted with the first agent or the second agent alone; and
administering the first agent to the subject wherein the output value that characterizes the change in cell adhesion or attachment is equal to or greater than 50%, indicating the first agent is therapeutically active in the cell signaling pathway of the subject's cancer cells.
2. The method of claim 1 , wherein the sample is contacted with the first agent and the second agent concurrently.
3. The method of claim 1 , wherein the sample is contacted with the first agent after contact with the second agent.
4. The method of claim 1 , wherein the sample is contacted with the second agent after contact with the first agent.
5. The method of claim 1 , wherein the signaling pathway is selected from the group consisting of MAPK, RHO, AKT, FAK1, RAS/RAF, PI3K/PTEN, MAK, MKK, and MEK.
6. The method of claim 1 , wherein the second agent is a protein, peptide, nucleic acid, metabolite, ligand, reagent, organic molecule, signaling factor, growth factor, biochemical, or combinations thereof.
7. The method of claim 1 , wherein cell adhesion or attachment is measured using an impedance biosensor or an optical biosensor.
8. The method of claim 1 , wherein the targeted therapeutic is one or more agents.
9. The method of claim 1 , wherein the change in cell adhesion or attachment is assessed using Euclidean analysis.
10. The method of claim 9 , 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.
11. The method of claim 1 , wherein the change in cell adhesion or attachment is measured by a change in temporal maxima or minima.
12. The method of claim 1 , wherein the targeted therapeutic is selected from the group consisting of cetuximab, docetaxel, erlotinib, gefitinib, irinotecan, lapatinib, paclitaxel, pazopanib, topotecan, trastuzumab, fulvestrant, tamoxifen, letrozole, anastrozole, exemestane, everolimus, abiraterone, bicalutamide, bortezomib, vemurafenib, ipilimumab and combinations thereof.
13. The method of claim 1 , wherein the cancer is selected from the group consisting of breast cancer, lung cancer, and colon cancer.
14. The method of claim 1 , wherein the change in cell adhesion or attachment is assessed using Euclidean analysis comprising arithmetic summation of the difference at multiple time points.
15. The method of claim 1 , wherein the sample of viable primary or metastatic cancer cells is also cultured in a media comprising growth factors and free of serum.
16. The method of claim 1 , wherein the sample of viable primary or metastatic cancer cells is also cultured in a media comprising an anti-apoptotic agent and free of serum.
17. A method of treating a human subject diagnosed with cancer, the method comprising:
administering to the subject a HER targeted therapeutic that has been determined to be therapeutically active in a HER family signaling pathway of the subject's cancer cells by a method comprising:
culturing a sample consisting essentially of viable primary or metastatic cancer cells obtained from the subject in a media free of serum;
contacting (1) a first portion of the sample with the HER targeted therapeutic and with neuregulin, and/or (2) contacting a second portion of the sample with the HER targeted therapeutic and with an epidermal growth factor;
continuously measuring cell adhesion or attachment of viable primary or metastatic cancer cells (1) in the first portion of the sample contacted with both the HER targeted therapeutic and neuregulin, relative to a sample of viable primary or metastatic cancer cells obtained from the subject which sample is contacted with the HER targeted therapeutic or neuregulin alone, and/or (2) in the second portion of the sample contacted with both the HER targeted therapeutic and an epidermal growth factor, relative to a sample of viable primary or metastatic cancer cells obtained from the subject which sample is contacted with the HER targeted therapeutic or an epidermal growth factor alone;
determining by mathematical analysis of the continuous measurements an output value, expressed as a percentage, that characterizes whether a change in cell adhesion or attachment has occurred (1) in the first portion contacted with both the HER targeted therapeutic and neuregulin, as compared to the sample contacted with the HER targeted therapeutic or neuregulin alone, and/or (2) in the second portion contacted with both the HER targeted therapeutic and an epidermal growth factor, as compared to the sample contacted with the HER targeted therapeutic or an epidermal growth factor alone; and
administering the HER targeted therapeutic to the subject wherein the output value that characterizes the change in cell adhesion or attachment is equal to or greater than 50%, indicating the HER targeted therapeutic is therapeutically active in the HER family signaling pathway of the subject's cancer cells.
18. The method of claim 17 , wherein the change in cell adhesion or attachment is assessed using Euclidean analysis.
19. The method of claim 18 , wherein the Euclidean analysis comprises arithmetic summation of the difference at multiple time points.
20. The method of claim 17 , wherein the sample of viable cells is also cultured in a media comprising growth factors and free of serum.
21. The method of claim 17 , wherein the sample of viable cells is also cultured in a media comprising an anti-apoptotic agent and free of serum.