Co-isolation of human donor eye cells and development of oncogene-mutated melanocytes to study uveal melanoma
Uveal melanoma (UM) is the most common intraocular tumor in adults, and its pathogenesis is only partly understood. UM can develop de novo from normal choroidal melanocytes (NCMs) or from pre-existing nevi that stem from NCMs and are thought to harbour UM-initiating mutations, most commonly in GNAQ or GNA11. However, there is a complete lack of commercially available NCM cell lines and no detailed protocol for developing oncogene-mutated CM line (MutCM) to study UM development. The purpose of this study was to establish and characterize premalignant CM models from human donor eyes to recapitulate the cell populations at the origin of UM. Overall design: We isolated NCMs from human donor eyes and generated MutCMs harboring one of the most common UM driver mutations (GNAQQ209L) by lentiviral transduction. To understand potential molecular changes induced by GNAQQ209L mutation, transcriptomic analyses comparing between a pair of NCM and MutCM derived from the same donor and between NCM and a uveal melanoma cell line (92.1) were performed.