title: Integration of genotypic and phenotypic screening reveals molecular mediators of melanoma-stromal interaction.
abstract: Tumor-endothelium interactions are critical for tumor survival and metastasis. Melanomas can rapidly metastasize early in tumor progression, but the dependence of this aggressive behavior on tumor-stromal interaction is poorly understood. To probe the mechanisms involved, we developed a heterotypic coculture methodology, allowing simultaneous tracking of genomic and phenotypic changes in interacting tumor and endothelial cells in vitro. We found a dramatic rearrangement of endothelial cell networks into patterns reminiscent of vascular beds, even on plastic and glass. Multiple genes were upregulated in the process, many coding for cell surface and secreted proteins, including Neuropilin-2 (NRP2). A critical role of NRP2 in coordinated cell patterning and growth was confirmed using the coculture system. We conclude that NRP2 represents an important mediator of melanoma-endothelial interactions. Furthermore, the described methodology represents a powerful yet simple system to elucidate heterotypic intercellular interactions mediating diverse physiological and pathological processes.
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Program in Cellular and Molecular Medicine, Department of Biomedical Engineering, Whitaker Institute for Biomedical Engineering, Institute for Cellular Engineering, and Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Stine
authors.forename: C Joanne
authors.initials: CJ
authors.lastname: Wang
authors.forename: Whei F
authors.initials: WF
authors.lastname: Moriarty
authors.forename: Byungwoo
authors.initials: B
authors.lastname: Ryu
authors.forename: Raymond
authors.initials: R
authors.lastname: Cheong
authors.forename: William H
authors.initials: WH
authors.lastname: Westra
authors.forename: Andre
authors.initials: A
authors.lastname: Levchenko
authors.forename: Rhoda M
authors.initials: RM
authors.lastname: Alani
mesh_terms: D002450:Cell Communication / Q000502:physiology*; D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D005838:Genotype; D006801:Humans; D008024:Ligands; D008545:Melanoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D039943:Neuropilin-2 / Q000096:biosynthesis / Q000235:genetics*; D010641:Phenotype; D017154:Stromal Cells / Q000473:pathology*; D015854:Up-Regulation
chemical_list: D008024:Ligands; D039943:Neuropilin-2