Whole cell assays and methods

• US 10,041,934 B2
• Filed: 06/24/2016
• Issued: 08/07/2018
• Est. Priority Date: 06/12/2012
• Status: Active Grant

• Current Assignee: Celcuity Inc.
• Original Assignee: Celcuity LLC
• Inventors: Laing, Lance Gavin, Sullivan, Brian Francis
• Primary Examiner(s): Dines, Karla A
• Application Number:
  US15/192,280
• Publication Number:
  US 20160305932A1
• Time in Patent Office: 774 Days
• Field of Search: 506 9
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: G01N 27/02 by investigating impedance
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity
• CPC Class Codes: G01N 33/5044 involving specific cell types
• CPC Class Codes: G01N 33/574 for cancer

Abstract

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.

Claims

1. A method of treating a human subject diagnosed with cancer, the method comprising:

administering to the subject a first agent that is a Human Epidermal growth factor Receptor (HER) family 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 comprising viable cancer cells obtained from the subject;

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 HER family targeted therapeutic is selected from a list consisting of neratinib, gefitinib, erlotinib, cetuximab, panitumumab, trastuzumab, lapatinib, and pertuzumab.

3. The method of claim 2, wherein the HER family targeted therapeutic is trastuzumab.

4. The method of claim 2, wherein the HER family targeted therapeutic is lapatinib.

5. The method of claim 2, wherein the HER family targeted therapeutic is pertuzumab.

6. The method of claim 2, wherein the HER family targeted therapeutic is cetuximab.

7. The method of claim 1, wherein the second agent is neuregulin.

8. The method of claim 1, wherein the second agent is heregulin.

9. The method of claim 1, wherein the second agent is an epidermal growth factor.

10. The method of claim 1, wherein cell adhesion or attachment is measured using an impedance biosensor or an optical biosensor.

11. The method of claim 1, wherein the change in cell adhesion or attachment is assessed using Euclidean analysis.

12. The method of claim 1, wherein the cancer is breast cancer.

13. The method of claim 1, wherein the sample is cultured in a media free of serum.

14. The method of claim 13, wherein the media free of serum further comprises at least one growth factor.

15. The method of claim 13, wherein the media free of serum further comprises at least one anti-apoptotic agent.

16. The method of claim 1, wherein the sample consists essentially of viable primary or metastatic cancer cells obtained from the subject and wherein the sample is cultured in a media free of serum.

17. The method of claim 16, wherein the media free of serum further comprises at least one growth factor.

18. The method of claim 16, wherein the media free of serum further comprises at least one anti-apoptotic agent.