Adenosine A2A Receptor Functional HEK293 Cell Line
Adenosine A2A receptor (A2aR or ADORA2A) stably expressed in HEK293 cells (NM_000675.5). A2aR is a member of the seven transmembrane G protein-coupled receptor (GPCR) family. The activity of A2aR is mediated by Gαs protein which activates adenylyl cyclase, resulting in the synthesis of intracellular cAMP. The level of cAMP correlates with the level of adenosine. This cell line can be used to measure the EC50 and IC50 values of A2aR agonists or antagonists.
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Media Required for Cell Culture
Name | Ordering Information |
Thaw Medium 1 | BPS Bioscience #60187 |
Growth Medium 1G | BPS Bioscience #79544 |
Materials Required for Cellular Assay
Name | Ordering Information |
Charcoal stripped fetal bovine serum | ThermoFisher #A3382101 |
IBMX | Sigma-Aldrich #I7018 |
Ro 20-1724 | Sigma Aldrich #557502 |
CGS-21680 hydrochloride hydrate | Sigma Aldrich #C141 |
ZM 241385 | Sigma Aldrich #Z0153 |
cAMP assay kit such as | |
cAMP-Gs Dynamic | PerkinElmer/Cisbio #62AM4PEB |
cAMPGlo kit | Promega #V1501 |
96-well PDL coated white clear-bottom assay plate | Corning #354651 |
Plate reader to read the cAMP assay kit |
The cell line has been screened to confirm the absence of Mycoplasma species.
Adenosine signaling plays an important role in inflammation and the immune response. Many cells in the tumor microenvironment express ectopic CD39 and CD73, leading to the buildup of extracellular adenosine. Engagement of adenosine with the high affinity Adenosine A2A receptor (A2aR) on the surface of T cells, macrophages, NK cells, neutrophils, and dendritic cells causes downregulation of the immune response. Therefore, A2aR is a novel immune checkpoint protein, and blockade of A2aR is being actively investigated as a potential immunotherapy. Several A2aR antagonists have progressed to clinical trials for the treatment of Parkinson’s disease, and preclinical studies have confirmed that blockade of A2aR activation has the ability to markedly enhance anti-tumor immunity. Mice treated with A2aR antagonists, such as ZM241385 or caffeine, show significantly delayed tumor growth, and A2aR knockout mice demonstrate increased tumor rejection. Most promising, A2aR blockade can be used in synergy with the inhibition of other immune checkpoint pathways. Studies show that the combination of A2aR blockade and PD-1 inhibition is more effective than either treatment separately, and A2aR blockade increases the activity of CTLA-4 and TIM-3 inhibition in controlling the growth of CD73+ melanoma.
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