Claims for Patent: 10,208,357

Summary for Patent: 10,208,357

Title:	Cell based assay
Abstract:	The present invention relates to cell-based assays involving HER2. The assays use assay cells that are transfected with cassettes containing the HER2 gene of interest and measure the effect of mutations on the activity of HER2, and on their response to inhibitors.
Inventor(s):	Yu; Yick Loi Raymond (East Brunswick, NJ)
Assignee:	Medical Diagnostic Laboratories, LLC (Hamilton, NJ)
Application Number:	15/464,520
Patent Claims:	1. A method of determining whether an HER2 variant is sensitive to treatment with a HER2 inhibitor in a cell, comprising the steps of: a) preparing a cDNA encoding said HER2 variant; b) preparing an expression cassette containing the HER2 variant cDNA; c) transfecting said prepared expression cassette containing said HER2 variant cDNA in an assay cell having a JNK reporter construct having SEQ ID NO: 31 comprising a reporter gene cDNA linked to at least one AP-1 binding site, and said cell is capable of expressing HER3, wherein the HER2 variant and HER3 are expressed; d) exposing said transfected cell to a HER3 ligand, wherein HER3 complexes with said HER2 variant to form a dimer which thereby activates the JNK reporter construct and generates a signal; e) exposing said transfected cell with an HER2 inhibitor; and f) determining whether said HER2 variant is sensitive to treatment with said HER2 inhibitor by measuring a change in signal. 2. The method of claim 1, wherein said HER2 variant contains a missense mutation, insertion, or deletion. 3. The method of claim 1 wherein the expression cassette containing the HER2 variant cDNA is a linear expression cassette. 4. The method of claim 1 wherein said assay cell has a double knockout of HER1 and HER2. 5. The method of claim 1 wherein the JNK reporter construct comprises a reporter gene cDNA linked to 3 to 12 AP-1 binding sites. 6. The method of claim 1 wherein the INK reporter construct comprises a reporter gene cDNA linked to 6 AP-1 binding sites. 7. The method of claim 1 wherein said assay cell further comprises a HER3 expression construct. 8. The method of claim 7 wherein said HER3 expression construct is stably integrated into the assay cell. 9. The method of claim 1 wherein said JNK reporter construct is stably integrated into the assay cell. 10. The method of claim 1, wherein step c) and step d) are performed simultaneously. 11. The method of claim 1 wherein steps a) through f) are performed in 32 hours or less. 12. The method of claim 1, wherein said HER2 inhibitor is Lapatinib, Trastuzumab, or Pertuzumab. 13. The method of claim 1, wherein said reporter gene cDNA encodes luciferase, said signal is light emission produced upon addition of a substrate for said luciferase, and a decrease in light emission relative to a control without HER2 inhibitor is indicative of the variant being sensitive to the inhibitor. 14. The method of claim 1 wherein said HER2 variant is obtained from a patient's biological sample consisting of blood, serum, and tumor tissue. 15. The method of claim 1, wherein said HER2 variant is transiently transfected. 16. A method of determining whether an HER2 variant is sensitive to treatment with an HER2 inhibitor in a cell, comprising the steps of: a) preparing a cDNA encoding said HER2 variant; b) preparing an expression cassette containing the HER2 variant cDNA; c) transfecting said prepared expression cassette containing said HER2 variant cDNA in an assay cell having a stably integrated JNK reporter construct having SEQ ID NO: 31 comprising a reporter gene cDNA linked to 6 AP-1 binding sites, said assay cell having a stably integrated HER3 expression construct; and said assay cell having a HER1 and HER2 double knockout, wherein the HER2 variant and HER3 are expressed; d) exposing said transfected cell to a HER activator, wherein HER3 complexes with said HER2 variant to form a dimer which thereby activates the JNK reporter construct and generates a signal; e) exposing said transfected cell with an HER2 inhibitor; and f) determining whether said HER2 variant is sensitive to treatment with said HER2 inhibitor by measuring a change in signal.

Details for Patent 10,208,357

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	09/25/1998		2039-02-26
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	02/10/2017		2039-02-26
Genentech, Inc.	PERJETA	pertuzumab	Injection	125409	06/08/2012		2039-02-26
Genentech, Inc.	HERCEPTIN HYLECTA	trastuzumab and hyaluronidase-oysk	Injection	761106	02/28/2019		2039-02-26