title: GPER-targeted, 99mTc-labeled, nonsteroidal ligands demonstrate selective tumor imaging and in vivo estrogen binding.
abstract: Our understanding of estrogen (17β-estradiol, E2) receptor biology has evolved in recent years with the discovery and characterization of a 7-transmembrane-spanning G protein-coupled estrogen receptor (GPER/GPR30) and the development of GPER-selective functional chemical probes. GPER is highly expressed in certain breast, endometrial, and ovarian cancers, establishing the importance of noninvasive methods to evaluate GPER expression in vivo. Here, we developed (99m)Tc-labeled GPER ligands to demonstrate the in vivo status of GPER as an estrogen receptor (ER) and for GPER visualization in whole animals. A series of (99m)Tc(I)-labeled nonsteroidal tetrahydro-3H-cyclopenta[c]quinolone derivatives was synthesized utilizing pyridin-2-yl hydrazine and picolylamine chelates. Radioligand receptor binding studies revealed binding affinities in the 10 to 30 nmol/L range. Cell signaling assays previously demonstrated that derivatives retaining a ketone functionality displayed agonist properties, whereas those lacking such a hydrogen bond acceptor were antagonists. In vivo biodistribution and imaging studies performed on mice bearing human endometrial and breast cancer cell xenografts yielded significant tumor uptake (0.4-1.1%ID/g). Blocking studies revealed specific uptake in multiple organs (adrenals, uterus, and mammary tissue), as well as tumor uptake with similar levels of competition by E2 and G-1, a GPER-selective agonist. In conclusion, we synthesized and evaluated a series of first-generation (99m)Tc-labeled GPER-specific radioligands, demonstrating GPER as an estrogen-binding receptor for the first time in vivo using competitive binding principles, and establishing the utility of such ligands as tumor imaging agents. These results warrant further investigation into the role of GPER in estrogen-mediated carcinogenesis and as a target for diagnostic/therapeutic/image-guided drug delivery.
  
  
  CONCLUSIONS
  These studies provide a molecular basis to evaluate GPER expression and function as an ER through in vivo imaging.
authors.affiliation: Department of Cell Biology and Physiology, School of Medicine, University of New Mexico Health Science Center, Albuquerque, New Mexico. College of Pharmacy, University of New Mexico Health Science Center, Albuquerque, New Mexico.
authors.forename: Tapan K
authors.initials: TK
authors.lastname: Nayak
authors.affiliation: Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico.
authors.forename: Chinnasamy
authors.initials: C
authors.lastname: Ramesh
authors.affiliation: Department of Cell Biology and Physiology, School of Medicine, University of New Mexico Health Science Center, Albuquerque, New Mexico. University of New Mexico Cancer Center, University of New Mexico Health Science Center, Albuquerque, New Mexico.
authors.forename: Helen J
authors.initials: HJ
authors.lastname: Hathaway
authors.affiliation: College of Pharmacy, University of New Mexico Health Science Center, Albuquerque, New Mexico. University of New Mexico Cancer Center, University of New Mexico Health Science Center, Albuquerque, New Mexico.
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Norenberg
authors.affiliation: Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico. University of New Mexico Cancer Center, University of New Mexico Health Science Center, Albuquerque, New Mexico.
authors.forename: Jeffrey B
authors.initials: JB
authors.lastname: Arterburn
authors.affiliation: Department of Cell Biology and Physiology, School of Medicine, University of New Mexico Health Science Center, Albuquerque, New Mexico. University of New Mexico Cancer Center, University of New Mexico Health Science Center, Albuquerque, New Mexico. eprossnitz@salud.unm.edu.
authors.forename: Eric R
authors.initials: ER
authors.lastname: Prossnitz
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D045744:Cell Line, Tumor; D003952:Diagnostic Imaging; D004967:Estrogens / Q000378:metabolism*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008024:Ligands; D008819:Mice, Nude; D009369:Neoplasms / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D010052:Ovariectomy; D015363:Quinolones / Q000737:chemistry; D011960:Receptors, Estrogen / Q000378:metabolism*; D013194:Staining and Labeling; D013667:Technetium; D013997:Time Factors; D014018:Tissue Distribution; D023041:Xenograft Model Antitumor Assays
chemical_list: D004967:Estrogens; D008024:Ligands; D015363:Quinolones; D011960:Receptors, Estrogen; D013667:Technetium