Store-operated calcium cellular assay
View Patent ↗The present technology provides a cell based assay for identifying compounds that modulate store-operated ionic calcium levels using itpr mutant cell lines, such as itpr-ku cells, which have abnormal ionic calcium levels.
1. A method for identifying a compound that modulates store-operated calcium entry (SOCE) levels in a cell, comprising:
contacting a mutant mammalian cell with a test compound,
wherein the mutant mammalian cell comprises a residual SOCE level and expresses a single 1,4,5-inositol triphosphate receptor (InsP 3 R) gene (itpr) product, wherein the itpr gene is an itpr ka1091/ug3 mutant, and wherein the itpr ka1091/ug3 product contains G1891S and S224F point mutations; and measuring a SOCE level in the test compound contacted cell,
wherein a measured increase or decrease of the SOCE level versus the residual SOCE level identifies the compound as a compound that modulates SOCE levels.
2. The method of claim 1 , further comprising:
measuring a SOCE level of a control cell and a SOCE level of the mutant mammalian cell; and
comparing the measured SOCE level of the mutant mammalian cell with the SOCE level of the control cell thereby determining the SOCE level difference between the control cell and mutant mammalian cell.
3. The method of claim 2 , wherein the control cell is selected from the group consisting of: (i) a wild-type, normal cell, and (ii) a cell having wild-type calcium ion release.
4. The method of claim 1 , wherein measuring a SOCE level comprises:
calcium imaging of the mutant mammalian cell and the test compound contacted mutant mammalian cell to obtain calcium imaging patterns and comparing the calcium imaging patterns before and after the contacting.
5. The method of claim 4 , further comprising:
comparing the calcium imaging patterns of the mutant mammalian cell with calcium imaging patterns of a control cell.
6. The method of claim 5 , wherein the control cell is selected from the group consisting of: (i) a wild-type, normal cell, and (ii) a cell having wild-type calcium ion release.
7. The method of claim 1 , wherein the test compound is a compound effective for the treatment of cells having abnormal SOCE levels in a disease characterized thereby.
8. The method of claim 7 , wherein the disease characterized by cells having abnormal SOCE levels is selected from the group consisting of spino-cerebellar ataxia (SCA), an immunodeficiency, severe combined immunodeficiency (SCID), Darier's disease (DAR), acute pancreatitis (AP), and Alzheimer's Disease (AD).