Methods of using lung or bronchial epithelial cells to identify bitter taste modulators

Patent metadata

Field	Value
Publication number	US-9128079-B2
Application number	US-201213569664-A
Country	US
Kind code	B2
Filing date	Aug 8, 2012
Priority date	Aug 8, 2011
Publication date	Sep 8, 2015
Grant date	Sep 8, 2015

Abstract

Provided herein are cell lines and assays that can be utilized to identify taste receptor modulators.

First claim

What is claimed is:

1. A method for identifying a candidate bitter taste modulator for potency testing comprising:
a) contacting a cell with a bitter tastant and a test compound, wherein the cell is a lung or bronchial epithelial cell derived from airway tissue and endogenously expresses a bitter taste receptor;
b) measuring bitter taste receptor activity, wherein a change in bitter taste receptor activity by the bitter tastant indicates modulation of the bitter taste receptor by the test compound, thus identifying a bitter taste modulator, and
c) identifying a candidate bitter taste modulator for potency testing.

2. The method of claim 1 , wherein the cell endogenously expresses Regulator of G-protein signaling-21 (RGS21).

3. The method of claim 1 , wherein the modulator inhibits the activity of the bitter tastant on the bitter taste receptor.

4. The method of claim 1 , wherein the bitter taste receptor is Taste receptor type 2, member 46 (T2R46) or Taste receptor type 2, member 38 (T2R38).

5. The method of claim 1 , wherein the cell is a 16HBE cell or derivative thereof.

6. The method of claim 1 , wherein the cell is modified to overexpress the bitter taste receptor.

7. The method of claim 1 , wherein bitter taste receptor activity is measured by detecting the level of an intracellular second messenger in the cell.

8. The method of claim 7 , wherein the second messenger is cAMP.

9. The method of claim 7 , wherein the second messenger is DAG or IP3.

10. The method of claim 1 , wherein bitter taste receptor activity is measured by detecting the level of intracellular calcium in the cell.

11. The method claim 1 , wherein the bitter taste receptor activity is binding activity.

12. The method of claim 11 , wherein a change in binding activity is detected by a competitive binding assay.

13. The method of claim 11 , wherein a change in binding activity is detected by surface plasmon resonance.

14. A method for identifying a bitter tastant comprising: a) contacting a 16HBE cell or derivative thereof with a test compound; b) measuring bitter taste receptor activity, wherein an increase in bitter taste receptor activity indicates that the test compound is a bitter tastant.

15. The method of claim 14 , wherein the bitter taste receptor is T2R46 or T2R38.

16. The method of claim 14 , wherein bitter taste receptor activity is measured by detecting the level of an intracellular second messenger in the cell.

17. The method of claim 16 , wherein the second messenger is cAMP.

18. The method of claim 16 , wherein the second messenger is DAG or IP3.

19. The method of claim 14 , wherein bitter taste receptor activity is measured by detecting the level of intracellular calcium in the cell.

20. The method of claim 14 , wherein the bitter taste receptor activity is binding activity.

21. The method of claim 20 , wherein binding activity is detected by a competitive binding assay.

22. The method of claim 20 , wherein a change in binding activity is detected by surface plasmon resonance.

23. The method of claim 14 , wherein the cell is modified to overexpress the bitter taste receptor.

Assignees

• Radhakrishna Harish
• Brown Michael D
• Siderovski David Peter
• Kimple Adam
• Cohen Staci Padove
• Coca Cola Co
• Univ North Carolina

Inventors

• Radhakrishna Harish
• Brown Michael D
• Siderovski David Peter
• Kimple Adam
• Cohen Staci Padove

Classifications

• C12N2503/02

  Drug screening · CPC title

• G01N33/5044 Primary

  involving specific cell types · CPC title