SCREENING ASSAYS BASED ON MAG AND/OR ABHD6 FOR SELECTING INSULIN SECRETION PROMOTING AGENT

摘要

The present application relates to a method of characterizing an agent's ability to increase insulin secretion in a subject. The method comprises determining whether the agent is able to modulate MAG level at the inner surface of the cytoplasmic membrane of a cell and/or ABHD6 activity. The agent is characterized as having the ability to increase insulin secretion in the subject when it is capable of upregulating MAG level at the inner surface of the cytoplasmic membrane and/or downregulating ABHD6 activity.

主权项

1. A method of characterizing an agent's ability to increase glucose-stimulated insulin secretion in a subject, said method comprising:
(a) combining the agent with a pancreatic β cell expressing a ABHD6 polypeptide, in the presence of at least 10 mM of glucose; (b) determining a test value of the lipase activity specific for the ABHD6 polypeptide of the pancreatic β cell in the presence of the agent and of glucose, wherein the test value is obtained by measuring, in the pancreatic β cell, a test level of intracellular monoacyl glycerol (MAG), a test level of extracellular glycerol and a test level of extracellular free fatty acid; (c) providing a control value of the lipase activity of the ABHD6 polypeptide, wherein the control value is associated with a lack of ability to increase insulin secretion in the subject and wherein the control value comprises a control level of intracellular MAG, a control level of extracellular glycerol and a control level of extracellular free fatty acid; (d) comparing the test value of step (b) to the control value of step (c); and (e) characterizing the agent as:
i. having the ability to increase glucose-stimulated insulin secretion in the subject when the test level of intracellular MAG is increased with respect to the control level of intracellular MAG, the test level of extracellular glycerol is decreased with respect to the control level of extracellular glycerol and the test level of extracellular free fatty acid is decreased with respect to the control level of extracellular free fatty acid; andii. lacking the ability to increase glucose-stimulated insulin secretion in the subject when the test level of intracellular MAG is decreased with respect to the control level of intracellular MAG, the test level of extracellular glycerol is increased with respect to the control level of extracellular glycerol or the test level of extracellular free fatty acid is increased with respect to the control level of extracellular free fatty acid.