Drug selection for gastric cancer therapy using antibody-based arrays

Patent metadata

Field	Value
Publication number	US-10473640-B2
Application number	US-201113014674-A
Country	US
Kind code	B2
Filing date	Jan 26, 2011
Priority date	Sep 21, 2006
Publication date	Nov 12, 2019
Grant date	Nov 12, 2019

Abstract

The present invention provides methods for selecting a suitable anticancer therapy, and for identifying and predicting response for the treatment of a gastric cancer.

First claim

What is claimed is:

1. A method for selecting a suitable anticancer drug for the treatment of a subject with gastric cancer, the method comprising:
(a) isolating a cancer cell from a subject with gastric cancer after administration of an anticancer drug, or prior to incubation with an anticancer drug;
(b) determining the activation level of at least each of the following analytes in a cellular extract produced from the isolated cancer cell: HER1, HER2, HER3, c-Met, IGF1R, and PI3K,
wherein determining the activation level of each of said analytes comprises:
(i) incubating the cellular extract with a dilution series of capture antibodies specific for each of said analytes to form a plurality of captured analytes, wherein the capture antibodies are restrained on a solid support;
(ii) washing and then incubating the plurality of captured analytes with detection antibodies comprising activation state-independent antibodies and activation state-dependent antibodies specific for each of said analytes to form a plurality of detectable captured analytes,
wherein the activation state-independent antibodies are labeled with glucose oxidase, wherein the glucose oxidase and the activation state-independent antibodies are conjugated to a sulfhydryl-activated dextran molecule, wherein the activation state-dependent antibodies are labeled with a first member of a signal amplification pair, and wherein the glucose oxidase generates an oxidizing agent which channels to and reacts with the first member of the signal amplification pair;
(iii) washing and then incubating the plurality of detectable captured analytes with a second member of the signal amplification pair to generate an amplified signal; and
(iv) detecting the amplified signal generated from the first and second members of the signal amplification pair;

(c) comparing the activation level of each of said analytes in the cellular extract to a reference activation level of each of said analytes that is generated in the absence of the anticancer drug; and
(d) selecting the anticancer drug as suitable for the treatment of the subject with gastric cancer when each of said analytes in the cellular extract is substantially less activated than in the absence of the anticancer drug.

2. The method of claim 1 , wherein the cancer cell is a circulating tumor cell or a fine needle aspirate (FNA) cell obtained from a tumor.

3. The method of claim 1 , wherein the cancer cell is isolated from a sample.

4. The method of claim 3 , wherein the sample is selected from the group consisting of whole blood, serum, plasma, tumor tissue, lymph, bone marrow aspirate, urine, saliva, and combinations thereof.

5. The method of claim 1 , wherein the anticancer drug is selected from the group consisting of a monoclonal antibody, tyrosine kinase inhibitor, anti-proliferative agent, chemotherapeutic agent, and combinations thereof.

6. The method of claim 5 , wherein the monoclonal antibody is selected from the group consisting of trastuzumab (Herceptin®), alemtuzumab (Campath®), bevacizumab (Avastin®), cetuximab (Erbitux®), gemtuzumab (Mylotarg®), panitumumab (Vectibix™), rituximab (Rituxan®), tositumomab (BEXXAR®), and combinations thereof.

7. The method of claim 5 , wherein the tyrosine kinase inhibitor is selected from the group consisting of gefitinib (Iressa®), sunitinib (Sutent®), erlotinib (Tarceva®), lapatinib (Tykerb®), canertinib (CI 1033), semaxinib (SU5416), vatalanib (PTK787/ZK222584), sorafenib (Nexavar®), imatinib mesylate (Gleevec®), leflunomide (SU101), and combinations thereof.

8. The method of claim 5 , wherein the anti-proliferative agent is an mTOR inhibitor selected from the group consisting of sirolimus (rapamycin), temsirolimus (CCI-779), everolimus (RAD001), and combinations thereof.

9. The method of claim 1 , wherein step (b) comprises determining both the expression level and activation level of each of said analytes.

10. The method of claim 1 , further comprising determining the expression level and/or activation level of one or more additional analytes in the cellular extract.

11. The method of claim 10 , wherein the one or more additional analytes comprises one or more signal transduction molecules selected from the group consisting of receptor tyrosine kinases, non-receptor tyrosine kinases, tyrosine kinase signaling cascade components, and combinations thereof.

12. The method of claim 10 , wherein the one or more additional analytes is selected from the group consisting of cKit, Shc, Akt, p70S6K, VEGFR, PDGFR, HER4, MEK, PTEN, SGK3, 4E-BP1, MAPK/ERK, PDK1, GSK-3β, Raf, SRC, NFkB-IkB, mTOR, Eph-a, Eph-b, Flt-3, Tie-1, Tie-2, Ab1, RET, FGFR1, FGFR2, FGFR3, FGFR4, RON, and combinations thereof.

13. The method of claim 1 , wherein the solid support is selected from the group consisting of glass, plastic, chips, pins, filters, beads, paper, membrane, fiber bundles, and combinations thereof.

14. The method of claim 1 , wherein the oxidizing agent is hydrogen peroxide (H 2 O 2 ).

15. The method of claim 14 , wherein the first member of the signal amplification pair is a peroxidase.

16. The method of claim 15 , wherein the peroxidase is horseradish peroxidase (HRP).

17. The method of claim 15 , wherein the second member of the signal amplification pair is a tyramide reagent.

18. The method of claim 1 , wherein the activation level is a phosphorylation level.

Assignees

• Singh Sharat
• Kim Phillip
• Liu Xinjun
• Ybarrondo Belen
• Nestle Sa

Inventors

• Singh Sharat
• Kim Phillip
• Liu Xinjun
• Ybarrondo Belen

Classifications

• G01N33/5011 Primary

  for testing antineoplastic activity · CPC title

• G01N33/5041

  involving analysis of members of signalling pathways · CPC title