title: Key components of the mode of action for hemangiosarcoma induction in pregabalin-treated mice: evidence of increased bicarbonate, dysregulated erythropoiesis, macrophage activation, and increased angiogenic growth factors in mice but not in rats.
abstract: In carcinogenicity studies, pregabalin increased hemangiosarcoma incidence in mice but not in rats. Investigative studies, ranging in length from 24 h to 12 months, were conducted in mice (1000 or 5000 mg/kg) and rats (900 mg/kg) to evaluate a potential mode-of-action scheme for tumor formation. Three areas were evaluated: (1) hematopoiesis (because endothelial and hematopoietic cells arise from the same precursor and hemangiosarcomas are primarily located in mouse hematopoietic tissues), (2) angiogenic growth factors (because increased angiogenic growth factors may stimulate vascular tumors), and (3) pulmonary/blood gas parameters (because hypoxia is a known driver for endothelial cell proliferation). In mice, pregabalin rapidly increased platelet and megakaryocyte counts, activated platelets and bone marrow erythrophages, decreased the myeloid-to-erythroid (M:E) ratio (49%), and produced bone marrow and splenic congestion and extramedullary hematopoiesis (EMH). Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor immunohistochemical staining were also increased in mouse bone marrow and spleen and vascular endothelial growth factor receptor 2 immunolabeling was increased in liver. Serum bicarbonate was increased within 24 h of pregabalin administration, persisted over time, and was accompanied by decreased respiratory rate (up to 34%) and increased partial pressure of carbon dioxide (pCO(2)), resulting in sustained metabolic alkalosis and elevated blood pH in mice. In contrast, in rats, pregabalin decreased overall bone marrow cellularity, including decreased number of megakaryocytes (24%) with no evidence of erythrophages, no change in M:E ratio, no EMH, and no increase in angiogenic growth factors or blood pH. Persistent alterations in serum bicarbonate, respiratory function, and blood gas parameters in mice, without adequate compensatory mechanisms, has the potential to create chronic tissue hypoxia, an accepted driver of endothelial cell proliferation.
authors.affiliation: Pfizer Worldwide Research and Development, Drug Safety Research and Development, Groton, Connecticut 06340, USA. kay.criswell@pfizer.com
authors.forename: Kay A
authors.initials: KA
authors.lastname: Criswell
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Wojcinski
authors.forename: David
authors.initials: D
authors.lastname: Pegg
authors.forename: Mudher
authors.initials: M
authors.lastname: Albassam
authors.forename: Steven
authors.initials: S
authors.lastname: Duddy
authors.forename: Eric
authors.initials: E
authors.lastname: Olsen
authors.forename: Marc
authors.initials: M
authors.lastname: Bailie
authors.forename: Stephen
authors.initials: S
authors.lastname: Foote
authors.forename: Timothy
authors.initials: T
authors.lastname: Anderson
mesh_terms: D000818:Animals; D001639:Bicarbonates / Q000378:metabolism*; D001784:Blood Gas Analysis; D004920:Erythropoiesis; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D006394:Hemangiosarcoma / Q000139:chemically induced* / Q000378:metabolism; D008262:Macrophage Activation / Q000187:drug effects*; D051379:Mice; D000069583:Pregabalin; D051381:Rats; D017208:Rats, Wistar; D012129:Respiratory Function Tests; D012189:Retrospective Studies; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000031:analogs & derivatives* / Q000633:toxicity
chemical_list: D001639:Bicarbonates; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; D000069583:Pregabalin; D005680:gamma-Aminobutyric Acid