Exploring the ATR-CHK1 pathway in the response of doxorubicin-induced DNA damages in acute lymphoblastic leukemia cells,Cell Biology and Toxicology

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Exploring the ATR-CHK1 pathway in the response of doxorubicin-induced DNA damages in acute lymphoblastic leukemia cells
Cell Biology and Toxicology ( IF 5.3 ) Pub Date : 2021-09-14 , DOI: 10.1007/s10565-021-09640-x
Andrea Ghelli Luserna Di Rorà ₁ , Martina Ghetti ₁ , Lorenzo Ledda ₁ , Anna Ferrari ₁ , Matteo Bocconcelli ₂ , Antonella Padella ₁ , Roberta Napolitano ₁ , Maria Chiara Fontana ₁ , Chiara Liverani ₁ , Enrica Imbrogno ₁ , Maria Teresa Bochicchio ₁ , Matteo Paganelli ₁ , Valentina Robustelli ₂ , Seydou Sanogo ₁ , Claudio Cerchione ₃ , Monica Fumagalli ₄ , Michela Rondoni ₅ , Annalisa Imovilli ₆ , Gerardo Musuraca ₃ , Giovanni Martinelli ₇ , Giorgia Simonetti ₁

Affiliation

Doxorubicin (Dox) is one of the most commonly used anthracyclines for the treatment of solid and hematological tumors such as B−/T cell acute lymphoblastic leukemia (ALL). Dox compromises topoisomerase II enzyme functionality, thus inducing structural damages during DNA replication and causes direct damages intercalating into DNA double helix. Eukaryotic cells respond to DNA damages by activating the ATM-CHK2 and/or ATR-CHK1 pathway, whose function is to regulate cell cycle progression, to promote damage repair, and to control apoptosis. We evaluated the efficacy of a new drug schedule combining Dox and specific ATR (VE-821) or CHK1 (prexasertib, PX) inhibitors in the treatment of human B−/T cell precursor ALL cell lines and primary ALL leukemic cells. We found that ALL cell lines respond to Dox activating the G2/M cell cycle checkpoint. Exposure of Dox-pretreated ALL cell lines to VE-821 or PX enhanced Dox cytotoxic effect. This phenomenon was associated with the abrogation of the G2/M cell cycle checkpoint with changes in the expression pCDK1 and cyclin B1, and cell entry in mitosis, followed by the induction of apoptosis. Indeed, the inhibition of the G2/M checkpoint led to a significant increment of normal and aberrant mitotic cells, including those showing tripolar spindles, metaphases with lagging chromosomes, and massive chromosomes fragmentation. In conclusion, we found that the ATR-CHK1 pathway is involved in the response to Dox-induced DNA damages and we demonstrated that our new in vitro drug schedule that combines Dox followed by ATR/CHK1 inhibitors can increase Dox cytotoxicity against ALL cells, while using lower drug doses.

Graphical abstract

• Doxorubicin activates the G2/M cell cycle checkpoint in acute lymphoblastic leukemia (ALL) cells.

• ALL cells respond to doxorubicin-induced DNA damages by activating the ATR-CHK1 pathway.

• The inhibition of the ATR-CHK1 pathway synergizes with doxorubicin in the induction of cytotoxicity in ALL cells.

• The inhibition of ATR-CHK1 pathway induces aberrant chromosome segregation and mitotic spindle defects in doxorubicin-pretreated ALL cells.

中文翻译：

探索 ATR-CHK1 通路在急性淋巴细胞白血病细胞阿霉素诱导的 DNA 损伤反应中的作用

阿霉素 (Dox) 是最常用的蒽环类药物之一，用于治疗实体瘤和血液肿瘤，例如 B-/T 细胞急性淋巴细胞白血病 (ALL)。 Dox 损害拓扑异构酶 II 酶的功能，从而在 DNA 复制过程中诱导结构损伤，并导致嵌入 DNA 双螺旋的直接损伤。真核细胞通过激活 ATM-CHK2 和/或 ATR-CHK1 通路来响应 DNA 损伤，其功能是调节细胞周期进程、促进损伤修复和控制细胞凋亡。我们评估了结合 Dox 和特定 ATR (VE-821) 或 CHK1 (prexasertib, PX) 抑制剂的新药物方案在治疗人 B-/T 细胞前体 ALL 细胞系和原代 ALL 白血病细胞中的疗效。我们发现所有细胞系都对激活 G2/M 细胞周期检查点的 Dox 做出反应。将 Dox 预处理的 ALL 细胞系暴露于 VE-821 或 PX 会增强 Dox 细胞毒性作用。这种现象与 G2/M 细胞周期检查点的废除、pCDK1 和细胞周期蛋白 B1 表达的变化以及细胞进入有丝分裂、随后诱导细胞凋亡有关。事实上，G2/M 检查点的抑制导致正常和异常有丝分裂细胞的显着增加，包括那些显示三极纺锤体、具有滞后染色体的中期和大量染色体碎片的细胞。总之，我们发现 ATR-CHK1 通路参与了对 Dox 诱导的 DNA 损伤的反应，并且我们证明了我们新的体外药物方案，结合了 Dox 和 ATR/CHK1 抑制剂，可以增加 Dox 对 ALL 细胞的细胞毒性，同时使用较低的药物剂量。