title: Cenp-E inhibitor GSK923295: Novel synthetic route and use as a tool to generate aneuploidy.
abstract: Aneuploidy is a common feature of cancer, with human solid tumour cells typically harbouring abnormal chromosome complements. The aneuploidy observed in cancer is often caused by a chromosome instability phenotype, resulting in genomic heterogeneity. However, the role aneuploidy and chromosome instability play in tumour evolution and chemotherapy response remains poorly understood. In some contexts, aneuploidy has oncogenic effects, whereas in others it is anti-proliferative and tumour-suppressive. Dissecting fully the role aneuploidy plays in tumourigenesis requires tools and facile assays that allow chromosome missegregation to be induced experimentally in cells that are otherwise diploid and chromosomally stable. Here, we describe a chemical biology approach that induces low-level aneuploidy across a large population of cells. Specifically, cells are first exposed to GSK923295, an inhibitor targeting the mitotic kinesin Cenp-E; while the majority of chromosomes align at the cell's equator, a small number cluster near the spindle poles. By then driving these cells into anaphase using AZ3146, an inhibitor targeting the spindle checkpoint kinase Mps1, the polar chromosomes are missegregated. This results in, on average, two chromosome missegregation events per division, and avoids trapping chromosomes in the spindle midzone, which could otherwise lead to DNA damage. We also describe an efficient route for the synthesis of GSK923295 that employs a novel enzymatic resolution. Together, the approaches described here open up new opportunities for studying cellular responses to aneuploidy.
authors: Faculty of Life Sciences, University of Manchester, United Kingdom.
authors: Ailsa
authors: A
authors: Bennett
authors: School of Chemistry, University of Manchester, United Kingdom.
authors: Beatrice
authors: B
authors: Bechi
authors: Faculty of Life Sciences, University of Manchester, United Kingdom.
authors: Anthony
authors: A
authors: Tighe
authors: Faculty of Life Sciences, University of Manchester, United Kingdom.
authors: Sarah
authors: S
authors: Thompson
authors: School of Chemistry, University of Manchester, United Kingdom.
authors: David J
authors: DJ
authors: Procter
authors: Faculty of Life Sciences, University of Manchester, United Kingdom.
authors: Stephen S
authors: SS
authors: Taylor
mesh_terms: D000782:Aneuploidy; D017209:Apoptosis; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000138:chemical synthesis / Q000737:chemistry*; D018797:Cell Cycle Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors*; D002875:Chromosomes / Q000648:ultrastructure; D004249:DNA Damage; D005434:Flow Cytometry; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins; D008938:Mitosis; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D034622:RNA Interference; D012521:Sarcosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D008941:Spindle Apparatus
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C571460:GSK923295; C068688:centromere protein E; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; C075840:TTK protein, human; D016547:Kinesins; D012521:Sarcosine
keywords: Cenp-E; Chromosome Section; Mps1; aneuploidy; chromosome instability; spindle assembly checkpoint