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

Tools used to assay genomic instability in cancers and cancer meiomitosis

“…Whereas numerical CIN, also known as aneuploidy, is characterized by a gain or loss of whole chromosomes that leads to a change in a cell karyotype (Tanaka and Hirota 2016). Defects underlying CIN include lagging chromosomes (Ricke et al 2008), micronuclei (He et al 2019), chromatin bridges (Passerini et al 2016), chromosome missegregation, cytokinesis failure (Fujiwara et al 2005; Nicholson et al 2015), telomere dysfunction (Gisselsson et al 2001) and spindle failure (Maiato and Logarinho 2014; Vitre et al 2015). Approaches to evaluate aneuploidy and examine defects underlying CIN remain critical in the study of cancer biology.…”

Natural antisense transcript of Period2, Per2AS, regulates the amplitude of the mouse circadian clock

“…Chromosomal numbers were counted as previous reported (Nicholson et al 2015) with minor modifications. Briefly, cell cultures were incubated in 1000 ng/mL Colcemid (Karyomax, Invitrogen) for 3-6 h at 37°C to enrich in mitotically arrested cells.…”

Cellular Stress Associated with Aneuploidy

“…The discrepancy could be attributed to differences in cell types or in the detection methods used. A recent study directly analyzed the frequency of anaphase lagging chromosomes in aneuploid and diploid cells (Nicholson et al, 2015). In the colorectal cancer cell line DLD-1, trisomy 7 or 13 cells were found to display higher frequencies of anaphase lagging chromosomes compared to the parental diploid cells.…”

Natural Antisense Transcript ofPeriod2, Per2AS, regulates the amplitude of the mouse circadian clock

“…Chromosomal number was counted as previous reported (Nicholson et al 2015) with minor modifications. Briefly, cell cultures were incubated in 1000 ng/ml Colcemid (Karyomax, Invitrogen) at 37°C for 3-6 hr to enrich in mitotically arrested cells.…”

CIN and Aneuploidy: Different Concepts, Different Consequences

“…It is important to keep in mind, that when one quantifies chromosome mis-segregation frequencies in tissue culture cells, most cell lines already exhibit low levels of chromosome missegregation, even in the absence of CIN-inducing mutations. [5,17,18,23,70] These mis-segregation events will lead to karyotype heterogeneity and karyotype evolution in the cell population and might explain the low rate of aneuploidy and karyotype evolution in cell cultures in which no (additional) CIN is induced (see Figure 2a for an example of karyotype evolution with low CIN). However, whether this "low-grade background" CIN rate is representative for the in vivo situation still needs to be assessed.…”

“…[14][15][16][17][18] In vitro CIN measurements can also be used to characterize animal models. For instance, some studies make use of mouse embryonic fibroblasts (MEFs) from mouse models of genetically induced CIN to estimate the in vivo rate of CIN.…”

“…[17,18] A trisomy of chromosome 13 in human cells can lead to failing cytokinesis, while a trisomy of chromosome 7 does not, showing that different aneuploid karyotypes can cause different forms of CIN. [17] However, in fully triploid fibroblasts (three copies of each chromosome) the percentage of aneuploid cells increases, but the rate of chromosome mis-segregation (CIN), relative to a euploid control, does not. [6] This can be explained by the fact that triploid cells that encountered chromosome losses had a growth advantage relative to their fully triploid counterparts, whereas chromosome losses were selected against in the diploid cells; this shows that the increased rate of aneuploidy was due to selection, not an increased CIN rate.…”

“…The effects of CIN have been found to be strongly dependent on the rate and type of chromosome mis-segregation or mitotic defect. [14][15][16][17][18] CIN is often measured in a tissue culture setting, which we, for the purpose of this review, define as in vitro. CIN can be quantified via time-lapse microscopy or immunofluorescence staining of fixed cells in mitosis.…”

“…[5,17,18,23,70] These mis-segregation events will lead to karyotype heterogeneity and karyotype evolution in the cell population and might explain the low rate of aneuploidy and karyotype evolution in cell cultures in which no (additional) CIN is induced (see Figure 2a for an example of karyotype evolution with low CIN). However, whether this "low-grade background" CIN rate is representative for the in vivo situation still needs to be assessed.…”

“…[40,41] Indeed, several trisomies, tetrasomies, and a few doubletrisomies were shown to cause further mis-segregation events in both cancer and non-cancer cell lines. [17,18] A trisomy of chromosome 13 in human cells can lead to failing cytokinesis, while a trisomy of chromosome 7 does not, showing that different aneuploid karyotypes can cause different forms of CIN. [17] However, in fully triploid fibroblasts (three copies of each chromosome) the percentage of aneuploid cells increases, but the rate of chromosome mis-segregation (CIN), relative to a euploid control, does not.…”

“…CIN is most often measured in vitro, for instance when assessing the effects of overexpression or inhibition of proteins or the effects of drugs. [14][15][16][17][18] In vitro CIN measurements can also be used to characterize animal models. For instance, some studies make use of mouse embryonic fibroblasts (MEFs) from mouse models of genetically induced CIN to estimate the in vivo rate of CIN.…”

“…It is difficult to make broad statements about CIN tolerance, as there are many different forms and rates of CIN. However, aneuploid cells with elevated levels of CIN [17] had a growth benefit relative to euploid control cells in non-standard, stressful culture conditions (i.e., serum-free conditions, and exposure to chemotherapeutic drug 5-Fluorouracil), but a growth defect under normal conditions. [69] This suggests that CIN may have selective advantages specifically in stressful, or non-standard growth conditions.…”

Small supernumerary marker chromosomes: A legacy of trisomy rescue?

“…The same applies to those sSMCs in which three different haplotypes at the level of the marker chromosome and biparental origin of the single nucleotide polymorphisms (SNPs) along the normal homologs are detected, with the only difference that the trisomic rescue occurred on one of the two chromosomes of maternal origin. It is well known that anaphase lagging accounts for trisomy rescue of the supernumerary chromosome (Ly & Cleveland, ; Nicholson et al., ) which is then trapped within a micronucleus where massive shattering occurs after disruption of the nuclear envelope exposing DNA to the cytoplasm (Liu et al., ; Zhang et al., ). As a consequence, the supernumerary chromosome is eliminated in one daughter cell, thus explaining the presence of the normal cell line.…”

The regulation of Net1/Cdc14 by the Hog1 MAPK upon osmostress unravels a new mechanism regulating mitosis

“…Mitotic exit is precisely and tightly coordinated to ensure that cell division occurs only after chromosomes are properly replicated and equally segregated between the two new daughter cells. Problems during mitotic exit can lead to genomic instability, genetic diseases and neurodegenerative disorders [133,134]. Various surveillance mechanisms or checkpoints delay mitotic progression to guarantee faithful inheritance of the genetic material.…”

Somatic Sex: On the Origin of Neoplasms With Chromosome Counts in Uneven Ploidy Ranges

“…To achieve this, membrane fragments first coat the individual chromosomes, which only then agglomerate and coalesce into distinct nuclei again (Schooley et al, 2012). Such single chromosome micronuclei form the basis for the microcell-mediated transfer of exogenous chromosome material into host cells (Meaburn et al, 2005;Fenech et al, 2011), which even enabled the successful integration of entire intact human chromosomes 8, 13, 18 and 21 into human isogenic embryonic stem cells (Kazuki et al, 2014;Hiramatsu et al, 2019) as well as chromosomes 3, 7 and 13 into the karyotypic stable, mismatch repair deficient colorectal cancer cell line DLD-1 (Upender et al, 2004;Nicholson et al, 2015;Rutledge and Cimini, 2016;Wangsa et al, 2019). A missegregated micronucleus can also undergo massive shattering and restructuring before these pieces rejoin again and form a new single chromatid, which can then become part of the nucleus again.…”

The Consequences of Chromosome Segregation Errors in Mitosis and Meiosis

“…In line with this hypothesis, yeast strains aneuploid in different chromosomes show various degrees of karyotypic and genomic instability, likely due to products from genes encoded on the aneuploid chromosomes [53,54]. Studies in human cells also suggest that aneuploidy itself may increase chromosome instability by affecting chromosome segregation in mitosis or by inducing defects in replication [55,56]. …”

Exploiting aneuploidy-imposed stresses and coping mechanisms to battle cancer

“…Similarly, aneuploid clones within human colorectal cancer cultures show a selective advantage and an increase in tumorigenic behaviour under stress conditions [20]. It has also been suggested that aneuploidy in a triploid or tetraploid cell can lead to further chromosomal instability, thereby promoting tumour evolution and tumorigenesis [30,31]. This process might well directly start after tetraploidization as the molecular machinery of tetraploid cells already displays molecular signatures that prepare cells for CIN tolerance.…”

“…For instance, lymphocytes from individuals born with systemic and stable trisomies for either chromosome 13, 18 and 21 show an increased frequency of aneuploidies for three other autosomes (chromosomes 8, 15 and 16) compared to lymphocytes of healthy controls suggesting that stable aneuploid cells tend to destabilize their genomes [101]. Similarly, DLD1 colorectal cancer cells carrying an extra chromosome 7 or 13 display reduced mitotic fidelity compared to diploid DLD1 cells [30], further suggesting that aneuploidy can induce chromosome missegregation.…”