HEK Cell Line Expressing Human EP1 Prostanoid Receptors

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John Regan
Professor, Pharmacology & Toxicology
Managed By
Tod Mccauley
Title: HEK Cell Line Expressing Human EP1 Prostaglandin Receptors
Basis of Invention: University of Arizona researchers have identified the EP1 subtype prostaglandin receptor from human cells, cloned the gene and created a cell line expressing the protein. Prostaglandins are known to regulate a variety of physiological and pathophysiological events in a wide range of tissues. The EP1 receptor has been specifically identified with mediating contractions of smooth muscles, including those in the gastrointestinal tract, the respiratory tract, the vas deferens, the myometrium and the sphincter of the iris. The receptors have also been implicated in regulating the adsorption of the small intestine and enhanced neuronal activity. The receptors are most abundant in the kidney, and are also commonly found in the lung, spleen and skeletal muscle, suggesting a range of other functions. Identification of the specific receptor protein and the production of cell lines expressing the protein are therefore expected to be major breakthroughs in understanding EP1 functions, and in developing drugs to cure diseases caused by malfunction of this receptor.
The Technology: The protein predicted to be EP1 receptor protein was cloned, and confirmed as the EP1 receptor. Xenopus oocytes injected with the putative EP1 cDNA showed a calcium influx when challenged with prostaglandin E2 (PGE2). EP1 had a stronger binding affinity for PGE2 than PGE1, which was greater than its affinity for PGF2a, which was greater than its affinity for PGD2. It responded to EP1 selective antagonists AH6809 and SC19220 much more potently than it responded to the EP2 receptor agonists. The researchers then stably transfected human embryonic kidney (HEK) cells with a eukaryotic expression vector containing the EP1 receptor cDNA isolated from kidney cells. They confirmed expression of the construct using antibodies.
Highlights: *              Cell lines can be used to look for drugs specifically interacting witht eh PG1 receptor *              Functional studies of receptor activation and second messenger signaling *              Sensitive and reliable measure of the potential for drugs to interact with the receptor. 
State of Development: Have identified and cloned the gene and protein, and produced stably transfected cell lines expressing it.