title: Effects of isoform-selective phosphatidylinositol 3-kinase inhibitors on osteoclasts: actions on cytoskeletal organization, survival, and resorption.
abstract: Phosphatidylinositol 3-kinases (PI3K) participate in numerous signaling pathways, and control distinct biological functions. Studies using pan-PI3K inhibitors suggest roles for PI3K in osteoclasts, but little is known about specific PI3K isoforms in these cells. Our objective was to determine effects of isoform-selective PI3K inhibitors on osteoclasts. The following inhibitors were investigated (targets in parentheses): wortmannin and LY294002 (pan-p110), PIK75 (α), GDC0941 (α, δ), TGX221 (β), AS252424 (γ), and IC87114 (δ). In addition, we characterized a new potent and selective PI3Kδ inhibitor, GS-9820, and explored roles of PI3K isoforms in regulating osteoclast function. Osteoclasts were isolated from long bones of neonatal rats and rabbits. Wortmannin, LY294002, GDC0941, IC87114, and GS-9820 induced a dramatic retraction of osteoclasts within 15-20 min to 65-75% of the initial area. In contrast, there was no significant retraction in response to vehicle, PIK75, TGX221, or AS252424. Moreover, wortmannin and GS-9820, but not PIK75 or TGX221, disrupted actin belts. We examined effects of PI3K inhibitors on osteoclast survival. Whereas PIK75, TGX221, and GS-9820 had no significant effect on basal survival, all blocked RANKL-stimulated survival. When studied on resorbable substrates, osteoclastic resorption was suppressed by wortmannin and inhibitors of PI3Kβ and PI3Kδ, but not other isoforms. These data are consistent with a critical role for PI3Kδ in regulating osteoclast cytoskeleton and resorptive activity. In contrast, multiple PI3K isoforms contribute to the control of osteoclast survival. Thus, the PI3Kδ isoform, which is predominantly expressed in cells of hematopoietic origin, is an attractive target for anti-resorptive therapeutics.
authors.affiliation: From the Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, Western University, London, Ontario N6A 5C1, Canada.
authors.forename: Ryan P P
authors.initials: RP
authors.lastname: Shugg
authors.forename: Ashley
authors.initials: A
authors.lastname: Thomson
authors.forename: Natsuko
authors.initials: N
authors.lastname: Tanabe
authors.forename: Adam
authors.initials: A
authors.lastname: Kashishian
authors.forename: Bart H
authors.initials: BH
authors.lastname: Steiner
authors.forename: Kamal D
authors.initials: KD
authors.lastname: Puri
authors.forename: Alexey
authors.initials: A
authors.lastname: Pereverzev
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Lannutti
authors.forename: Frank R
authors.initials: FR
authors.lastname: Jirik
authors.forename: S Jeffrey
authors.initials: SJ
authors.lastname: Dixon
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Sims
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001862:Bone Resorption / Q000517:prevention & control; D002470:Cell Survival / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects; D007191:Indazoles / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D009025:Morpholines / Q000494:pharmacology; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology; D053245:RANK Ligand / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D013449:Sulfonamides / Q000494:pharmacology; D000077191:Wortmannin
chemical_list: C532162:2-(1H-indazol-4-yl)-6-(4-methanesulfonylpiperazin-1-ylmethyl)-4-morpholin-4-ylthieno(3,2-d)pyrimidine; D000730:Androstadienes; D002867:Chromones; C477872:IC 87114; D007191:Indazoles; D007527:Isoenzymes; D009025:Morpholines; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D053245:RANK Ligand; D013449:Sulfonamides; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D000225:Adenine; D000077191:Wortmannin
keywords: Actin; Bone; Bone Resorption; Cytoskeleton; GS-9820; Lamellipod; Osteoclast; Phosphatidylinositol 3-Kinase