title: Peginesatide and erythropoietin stimulate similar erythropoietin receptor-mediated signal transduction and gene induction events.
abstract: Peginesatide is a synthetic, PEGylated, peptide-based erythropoiesis-stimulating agent that is designed and engineered to stimulate specifically the erythropoietin receptor dimer that governs erythropoiesis. Peginesatide has a unique structure that consists of a synthetic peptide dimer (with no sequence similarity to erythropoietin) conjugated to a 40-kDa PEG moiety. Peginesatide is being developed for the treatment of anemia associated with chronic kidney disease in dialysis patients. To compare signaling effects of peginesatide to recombinant human erythropoietin (rHuEPO), dose-dependent effects on protein phosphorylation and gene expression were evaluated using phosphoproteomics, quantitative signal transduction analyses, and gene profiling. After stimulation with peginesatide or rHuEPO, cell lysates were prepared from UT-7/EPO cells. Liquid chromatography-tandem mass spectrometry and MesoScale arrays were used to quantify phosphorylation events. Transcriptional changes were analyzed using microarrays and quantitative reverse transcription polymerase chain reaction. Peginesatide and rHuEPO were found to regulate the tyrosine phosphorylation of an essentially equivalent set of protein substrates, and modulate the expression of a similar set of target genes. Consistent with their roles in stimulating erythropoiesis, peginesatide and rHuEPO regulate similar cellular pathways.
authors.affiliation: Affymax, Inc., Palo Alto, CA, USA. jennifer_green@affymax.com
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Green
authors.forename: Karen
authors.initials: K
authors.lastname: Leu
authors.forename: Angela
authors.initials: A
authors.lastname: Worth
authors.forename: Richard B
authors.initials: RB
authors.lastname: Mortensen
authors.forename: David K
authors.initials: DK
authors.lastname: Martinez
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Schatz
authors.forename: Don M
authors.initials: DM
authors.lastname: Wojchowski
authors.forename: Peter R
authors.initials: PR
authors.lastname: Young
mesh_terms: D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004920:Erythropoiesis / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D010455:Peptides / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D017467:Receptors, Erythropoietin / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: C554270:EPO protein, human; D010455:Peptides; D017467:Receptors, Erythropoietin; D004921:Erythropoietin; C556270:peginesatide