Chromosome mis-segregation and cytokinesis failure in trisomic human cells

Abstract

Cancer cells display aneuploid karyotypes and typically mis-segregate chromosomes at high rates, a phenotype referred to as chromosomal instability (CIN). To test the effects of aneuploidy on chromosome segregation and other mitotic phenotypes we used the colorectal cancer cell line DLD1 (2n = 46) and two variants with trisomy 7 or 13 (DLD1+7 and DLD1+13), as well as euploid and trisomy 13 amniocytes (AF and AF+13). We found that trisomic cells displayed higher rates of chromosome mis-segregation compared to their euploid counterparts. Furthermore, cells with trisomy 13 displayed a distinctive cytokinesis failure phenotype. We showed that up-regulation of SPG20 expression, brought about by trisomy 13 in DLD1+13 and AF+13 cells, is sufficient for the cytokinesis failure phenotype. Overall, our study shows that aneuploidy can induce chromosome mis-segregation. Moreover, we identified a trisomy 13-specific mitotic phenotype that is driven by up-regulation of a gene encoded on the aneuploid chromosome.DOI: http://dx.doi.org/10.7554/eLife.05068.001

A Markov chain for numerical chromosomal instability in clonally expanding populations

“…It is shown in [ 20 ] that certain karyotypes promote cytokinesis failure and thus genome duplication. In particular, it is suggested that cells with 3 or more copies of chromosome 13 have a higher genome duplication rate.…”

High rates of chromosome missegregation suppress tumor progression but do not inhibit tumor initiation

“…Human chromosomally stable colorectal cancer cell lines (DLD1) with an extra copy of chromosome 7 or 13 show an increase in chromosome missegregation rates, as do amniotic fibroblasts that are trisomic for chromosome 13. Of interest, these cells predominantly missegregate the trisomic chromosome, suggesting that a portion of them revert to the diploid state ( Nicholson et al. , 2015 ).…”

Aneuploidy Causes Non-genetic Individuality

“…Although aneuploid cells are genomically unstable (Blank et al, 2015; Nicholson et al, 2015; Passerini et al, 2016; Sheltzer et al, 2011; Zhu et al, 2012), mutation-induced population heterogeneity is an unlikely cause. The system we employ examines the immediate effects of mis-segregating a specific chromosome, precluding the accumulation of suppressor mutations.…”

Engineering of Systematic Elimination of a Targeted Chromosome in Human Cells

“…An increased rate of gain or loss of whole chromosomes during each cell cycle is a common feature of many tumor cells, referred to as chromosome instability [12, 13]. Moreover, the rate of chromosome missegregation is higher in trisomic human cells than in euploid cells [14]. Therefore, extra copies of chromosome 21 integrating the DNA cassette may tend to be lost from the HeLa cells spontaneously under condition in which cells including HSV-tk gene are excluded.…”

Chromosomal localization of Ewing sarcoma EWSR1/FLI1 protein promotes the induction of aneuploidy

“…1 A ) ( 29 , 33 ). The DLD-1 colorectal cancer cell line was chosen because it does not express EWSR1/FLI1 endogenously, and because it has a near-diploid karyotype, it has been used as a model to evaluate proteins of interest for their ability to induce aneuploidy ( 24 , 34 ). The inducible system enabled us to assess the activity of EWSR1/FLI1 in the induction of aneuploidy after a single cell cycle.…”

“…Using these three DNA constructs, we established stable Tet-on cell lines by integrating EWSR1/FLI1, EWSR1/FLI1-T79A and EWSR1/FLI1-T79D transgenes (each with an mCherry tag) into the safe-harbor AAVS1 locus of DLD-1 cells using CRISPR/Cas9 technology (Fig 1A) (29,33). The DLD-1 colorectal cancer cell line was chosen because it does not express EWSR1/FLI1 endogenously, and, because it has a near-diploid karyotype, it has been used as a model to evaluate proteins of interest for their ability to induce aneuploidy (24,34). The inducible system enabled us to assess the activity of EWSR1/FLI1 in the induction of aneuploidy after a single cell cycle.…”

“…Although the tumor cells display a high incidence of aneuploidy, it is unclear whether aneuploidy is induced in the early stages of tumorigenesis and promotes the progression of the disease, or if it is a by-product of tumorigenesis and expands during tumor progression. Aneuploidy is the result of abnormal chromosome segregation, which may be caused by the failure of any of several mitotic processes (including chromosome cohesion; mitotic spindle (K-fiber) attachment to the kinetochore of the chromosome; the spindle assembly checkpoint (SAC); the amplification of centrosomes; or cytokinesis accompanied by defects in midbody structure) ( 23 , 24 ). We previously demonstrated that EWSR1/FLI1 impairs the recruitment of the key mitotic regulator Aurora B kinase to the midzone, (the network of antiparallel microtubules and midzone proteins located between segregating chromosomes during anaphase) ( 25 ).…”

MUN (MERISTEM UNSTRUCTURED), encoding a SPC24 homolog of NDC80 kinetochore complex, affects development through cell division in Arabidopsis thaliana

“…In other respects, incomplete or delayed cell plate formation in the mun‐1 mutant by mis‐segregation of chromosomes during the process of cell division may affect mis‐localization of PIN1, which perturbs proper auxin flow. In humans, cancer cells that display aneuploid karyotypes and mis‐segregate chromosomes show a distinctive cytokinesis failure phenotype (Nicholson et al ., ). Because the growth of the cell plate is dependent upon the phragmoplast formation in plants (Higaki et al ., ) and chromosome defects generally affect phragmoplast development, we expected the defects in cell walls in mun‐1 .…”

Trisomy 21-associated increases in chromosomal instability are unmasked by comparing isogenic trisomic/disomic leukocytes from people with mosaic Down syndrome

“…Aneuploidy has been posited to predispose somatic cells to acquired chromosomal instability [30][31][32][33][34][35], but this hypothesis has been challenging to test in humans [36]. Most of the extant literature assessing aneuploidy-related SCINF has been derived from cancer cell lines, the latter of which often have a cascade of biological/genetic changes that can confound the interpretation of the resultant data.…”

“…Passerini, et al [46] further suggested that the observed increase in trisomy-related genomic instability was attributable to replication defects and increased sensitivity to replication stress. Other investigators have also reported increases in SCINF in trisomic cells, with several of these investigators attributing the increase in chromosomal instability/mis-segregation to anaphase lagging of a chromosome or chromatid [32,[47][48][49].…”

Comparative proximity biotinylation implicates RAB18 in sterol mobilization and biosynthesis

“…More ambiguously, the RAB18-interacting microtubule binding proteins have not previously been reported to work together but do function in compatible locations. SPG20 and CAMSAP1 each associate with mitotic spindle poles, REEP4 participates in spindle-dependent ER clearance from metaphase chromatin and BICD2 is a component of the minus-end-directed dynein-dynactin motor complex (Hendershott & Vale, 2014, Hueschen et al, 2017, Kumar et al, 2019, Lind et al, 2011, Nicholson et al, 2015, Schlaitz et al, 2013, Urnavicius et al, 2015). Our TRAPPII-dependent RAB18 interaction data indicate that different GEF complexes affect largely distinct subsets of interactions.…”

Environmental stresses induce karyotypic instability in colorectal cancer cells

“…, 2009; Sheltzer et al. , 2011; Nicholson et al. , 2015) suggests that even a transient exposure to microenvironmental stresses could be sufficient to kick-start a vicious loop, generating further genetic and karyotypic variation.…”

“…To support this notion, the presence of specific aneuploid chromosomes has been shown to increase genomic instability (Zhu et al. , 2012; Nicholson et al. , 2015) and drug resistance (Lee et al.…”

Near‐tetraploid cancer cells show chromosome instability triggered by replication stress and exhibit enhanced invasiveness

“…It is now generally accepted that CIN and aneuploidy are fundamental forces driving carcinogenesis (23). Additionally, aneuploidy per se has been shown to promote chromosome missegregation in both yeast (24,25) and mammalian cells (26,27). Also, tetraploidy has been proposed as a possible precursor of aneuploidy in cancer (2).…”