title: High-throughput screening identifies idarubicin as a preferential inhibitor of smooth muscle versus endothelial cell proliferation.
abstract: Intimal hyperplasia is the cause of the recurrent occlusive vascular disease (restenosis). Drugs currently used to treat restenosis effectively inhibit smooth muscle cell (SMC) proliferation, but also inhibit the growth of the protective luminal endothelial cell (EC) lining, leading to thrombosis. To identify compounds that selectively inhibit SMC versus EC proliferation, we have developed a high-throughput screening (HTS) format using human cells and have employed this to screen a multiple compound collection (NIH Clinical Collection). We developed an automated, accurate proliferation assay in 96-well plates using human aortic SMCs and ECs. Using this HTS format we screened a 447-drug NIH Clinical Library. We identified 11 compounds that inhibited SMC proliferation greater than 50%, among which idarubicin exhibited a unique feature of preferentially inhibiting SMC versus EC proliferation. Concentration-response analysis revealed this differential effect most evident over an ∼10 nM-5 µM window. In vivo testing of idarubicin in a rat carotid injury model at 14 days revealed an 80% reduction of intimal hyperplasia and a 45% increase of lumen size with no significant effect on re-endothelialization. Taken together, we have established a HTS assay of human vascular cell proliferation, and identified idarubicin as a selective inhibitor of SMC versus EC proliferation both in vitro and in vivo. Screening of larger and more diverse compound libraries may lead to the discovery of next-generation therapeutics that can inhibit intima hyperplasia without impairing re-endothelialization.
authors.affiliation: Department of Surgery, University of Wisconsin, Madison, Wisconsin, United States of America.
authors.forename: Shakti A
authors.initials: SA
authors.lastname: Goel
authors.affiliation: Department of Surgery, University of Wisconsin, Madison, Wisconsin, United States of America.
authors.forename: Lian-Wang
authors.initials: LW
authors.lastname: Guo
authors.affiliation: Department of Surgery, University of Wisconsin, Madison, Wisconsin, United States of America.
authors.forename: Bowen
authors.initials: B
authors.lastname: Wang
authors.affiliation: Small Molecule Screening & Synthesis Facility, UW Carbone Cancer Center, Madison, Wisconsin, United States of America.
authors.forename: Song
authors.initials: S
authors.lastname: Guo
authors.affiliation: Department of Surgery, University of Wisconsin, Madison, Wisconsin, United States of America.
authors.forename: Drew
authors.initials: D
authors.lastname: Roenneburg
authors.affiliation: Small Molecule Screening & Synthesis Facility, UW Carbone Cancer Center, Madison, Wisconsin, United States of America.
authors.forename: Gene E
authors.initials: GE
authors.lastname: Ananiev
authors.affiliation: Small Molecule Screening & Synthesis Facility, UW Carbone Cancer Center, Madison, Wisconsin, United States of America.
authors.forename: F Michael
authors.initials: FM
authors.lastname: Hoffmann
authors.affiliation: Department of Surgery, University of Wisconsin, Madison, Wisconsin, United States of America.
authors.forename: K Craig
authors.initials: KC
authors.lastname: Kent
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D006965:Hyperplasia / Q000188:drug therapy; D015255:Idarubicin / Q000494:pharmacology*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D017539:Tunica Intima / Q000187:drug effects*
chemical_list: D015255:Idarubicin