复制应激导致人类诱导多能干细胞中ANKS1B大神经元基因的有丝分裂进入延迟和12号染色体脆性。

IF 2.4 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Anastasiia V Kislova, Diana Zheglo, Victoria O Pozhitnova, Philipp S Sviridov, Elmira P Gadzhieva, Ekaterina S Voronina
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引用次数: 1

摘要

大量背景水平的复制应激是胚胎和诱导多能干细胞(iPSC)的一个特征,它可能导致染色体数量和结构的不稳定,包括12号染色体的复发性畸变。在分化的细胞中,复制应激敏感的基因组区域,包括常见的脆性位点,通过温和的阿西德林处理(一种复制聚合酶的抑制剂)诱导有丝分裂染色体断裂而被广泛定位。IPSCs表现出较低的凋亡阈值和较高的修复能力,阻碍了脆性位点的定位。咖啡因增强基因毒性作用,消除化学和物理诱变剂诱导的G2/M检查点延迟。使用5-乙炔基-2'-脱氧尿苷(EdU)进行复制标记,我们表征了暴露于阿片苷和/或咖啡因的异步iPSC的有丝分裂进入动力学。在复制应激暴露时间调整后,细胞周期延迟,与原代淋巴细胞相比,在iPSC中观察到更高的中期染色体断裂率。使用差异染色体染色和随后的位点特异性荧光原位杂交,我们在12q23.1定位了横跨大神经元ANKS1B基因的FRA12L脆性位点,这可能有助于iPSC中复发性的12号染色体错序和重排。综述了关于ANKS1B基因改变及其可能的功能影响的公开可用数据。我们的研究首次证明了iPSC中常见的脆性位点诱导,并揭示了在人类早期发育和体外细胞扩增过程中临床相关基因的潜在体细胞不稳定性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Replication stress causes delayed mitotic entry and chromosome 12 fragility at the ANKS1B large neuronal gene in human induced pluripotent stem cells.

Replication stress causes delayed mitotic entry and chromosome 12 fragility at the ANKS1B large neuronal gene in human induced pluripotent stem cells.

Substantial background level of replication stress is a feature of embryonic and induced pluripotent stem cells (iPSCs), which can predispose to numerical and structural chromosomal instability, including recurrent aberrations of chromosome 12. In differentiated cells, replication stress-sensitive genomic regions, including common fragile sites, are widely mapped through mitotic chromosome break induction by mild aphidicolin treatment, an inhibitor of replicative polymerases. IPSCs exhibit lower apoptotic threshold and higher repair capacity hindering fragile site mapping. Caffeine potentiates genotoxic effects and abrogates G2/M checkpoint delay induced by chemical and physical mutagens. Using 5-ethynyl-2'-deoxyuridine (EdU) for replication labeling, we characterized the mitotic entry dynamics of asynchronous iPSCs exposed to aphidicolin and/or caffeine. Under the adjusted timing of replication stress exposure accounting revealed cell cycle delay, higher metaphase chromosome breakage rate was observed in iPSCs compared to primary lymphocytes. Using differential chromosome staining and subsequent locus-specific fluorescent in situ hybridization, we mapped the FRA12L fragile site spanning the large neuronal ANKS1B gene at 12q23.1, which may contribute to recurrent chromosome 12 missegregation and rearrangements in iPSCs. Publicly available data on the ANKS1B genetic alterations and their possible functional impact are reviewed. Our study provides the first evidence of common fragile site induction in iPSCs and reveals potential somatic instability of a clinically relevant gene during early human development and in vitro cell expansion.

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来源期刊
Chromosome Research
Chromosome Research 生物-生化与分子生物学
CiteScore
4.70
自引率
3.80%
发文量
31
审稿时长
1 months
期刊介绍: Chromosome Research publishes manuscripts from work based on all organisms and encourages submissions in the following areas including, but not limited, to: · Chromosomes and their linkage to diseases; · Chromosome organization within the nucleus; · Chromatin biology (transcription, non-coding RNA, etc); · Chromosome structure, function and mechanics; · Chromosome and DNA repair; · Epigenetic chromosomal functions (centromeres, telomeres, replication, imprinting, dosage compensation, sex determination, chromosome remodeling); · Architectural/epigenomic organization of the genome; · Functional annotation of the genome; · Functional and comparative genomics in plants and animals; · Karyology studies that help resolve difficult taxonomic problems or that provide clues to fundamental mechanisms of genome and karyotype evolution in plants and animals; · Mitosis and Meiosis; · Cancer cytogenomics.
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