title: Phenotype-based high-content chemical library screening identifies statins as inhibitors of in vivo lymphangiogenesis.
abstract: Lymphangiogenesis plays an important role in promoting cancer metastasis to sentinel lymph nodes and beyond and also promotes organ transplant rejection. We used human lymphatic endothelial cells to establish a reliable three-dimensional lymphangiogenic sprouting assay with automated image acquisition and analysis for inhibitor screening. This high-content phenotype-based assay quantifies sprouts by automated fluorescence microscopy and newly developed analysis software. We identified signaling pathways involved in lymphangiogenic sprouting by screening the Library of Pharmacologically Active Compounds (LOPAC)(1280) collection of pharmacologically relevant compounds. Hit characterization revealed that mitogen-activated protein kinase kinase (MEK) 1/2 inhibitors substantially block lymphangiogenesis in vitro and in vivo. Importantly, the drug class of statins, for the first time, emerged as potent inhibitors of lymphangiogenic sprouting in vitro and of corneal and cutaneous lymphangiogenesis in vivo. This effect was mediated by inhibition of the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase and subsequently the isoprenylation of Rac1. Supplementation with the enzymatic products of HMG-CoA reductase functionally rescued lymphangiogenic sprouting and the recruitment of Rac1 to the plasma membrane.
authors.affiliation: Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, Eidgenössische Technische Hochschule Zurich, Wolfgang-Pauli-Strasse 10, 8093 Zurich, Switzerland.
authors.forename: Martin Michael Peter
authors.initials: MM
authors.lastname: Schulz
authors.forename: Felix
authors.initials: F
authors.lastname: Reisen
authors.forename: Silvana
authors.initials: S
authors.lastname: Zgraggen
authors.forename: Stephanie
authors.initials: S
authors.lastname: Fischer
authors.forename: Don
authors.initials: D
authors.lastname: Yuen
authors.forename: Gyeong Jin
authors.initials: GJ
authors.lastname: Kang
authors.forename: Lu
authors.initials: L
authors.lastname: Chen
authors.forename: Gisbert
authors.initials: G
authors.lastname: Schneider
authors.forename: Michael
authors.initials: M
authors.lastname: Detmar
mesh_terms: D000704:Analysis of Variance; D042783:Endothelial Cells / Q000187:drug effects*; D005455:Fluorescent Antibody Technique; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D042583:Lymphangiogenesis / Q000187:drug effects*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010641:Phenotype; D015398:Signal Transduction / Q000187:drug effects*