A survey of chromosomal instability measures across mechanistic models.
A survey of chromosomal instability measures across mechanistic models.
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跨机械模型的染色体不稳定性测量的调查。
DOI：
10.1073/pnas.2309621121
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发表时间：
2024
影响因子：
11.1
通讯作者：
Burkard,MarkE
中科院分区：
文献类型：
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作者：
Lynch,AndrewR;Bradford,Shermineh;Zhou,AmberS;Oxendine,Kim;Henderson,Les;Horner,VanessaL;Weaver,BethA;Burkard,MarkE
染色体不稳定性（CIN）是在细胞分裂过程中通过染色体错误分离导致的癌症核型的持续重排。在癌症中，CIN以不同的水平存在，对肿瘤进展具有不同的影响。然而，尽管有一系列可用的措施，但在人类癌症中评估错误分离率仍然具有挑战性。我们通过比较使用染色体桥、假双极纺锤体、多极纺锤体和极性染色体的特异性、诱导型CIN模型的定量方法来评估CIN的测量。对于每一个，我们测量了CIN固定和延时荧光显微镜，染色体扩散，六着丝粒FISH，批量转录组学和单细胞DNA测序（scDNAseq）。正如预期的那样，活样本和固定样本中肿瘤细胞的显微镜检查显著相关（R = 0.72;P< 0.001），并且灵敏地检测CIN。细胞遗传学方法包括染色体扩散和6-着丝粒FISH，其也显著相关（R = 0.76;P< 0.001），但对于较低的CIN率具有有限的敏感性。批量基因组DNA特征和批量转录组学评分CIN 70和HET 70未检测到CIN。相比之下，scDNAseq以高灵敏度检测CIN，并且与成像方法显著相关（R = 0.82;P< 0.001）。总之，单细胞方法，如成像，细胞遗传学和scDNAseq可以测量CIN，后者是临床样本可获得的最全面的方法。为了便于比较表型和方法之间的CIN率，我们提出了CIN的标准化单位：每个二倍体分裂的错误分离。对常见CIN测量的系统分析突出了单细胞方法的优越性，并为临床环境中测量CIN提供了指导。
Chromosomal instability (CIN) is the persistent reshuffling of cancer karyotypes via chromosome mis-segregation during cell division. In cancer, CIN exists at varying levels that have differential effects on tumor progression. However, mis-segregation rates remain challenging to assess in human cancer despite an array of available measures. We evaluated measures of CIN by comparing quantitative methods using specific, inducible phenotypic CIN models of chromosome bridges, pseudobipolar spindles, multipolar spindles, and polar chromosomes. For each, we measured CIN fixed and timelapse fluorescence microscopy, chromosome spreads, six-centromere FISH, bulk transcriptomics, and single-cell DNA sequencing (scDNAseq). As expected, microscopy of tumor cells in live and fixed samples significantly correlated (R = 0.72;P< 0.001) and sensitively detect CIN. Cytogenetics approaches include chromosome spreads and 6-centromere FISH, which also significantly correlate (R = 0.76;P< 0.001) but had limited sensitivity for lower rates of CIN. Bulk genomic DNA signatures and bulk transcriptomic scores, CIN70 and HET70, did not detect CIN. By contrast, scDNAseq detects CIN with high sensitivity, and significantly correlates with imaging methods (R = 0.82;P< 0.001). In summary, single-cell methods such as imaging, cytogenetics, and scDNAseq can measure CIN, with the latter being the most comprehensive method accessible to clinical samples. To facilitate the comparison of CIN rates between phenotypes and methods, we propose a standardized unit of CIN: Mis-segregations per Diploid Division. This systematic analysis of common CIN measures highlights the superiority of single-cell methods and provides guidance for measuring CIN in the clinical setting.