The PPP2R1A cancer hotspot mutant p.R183W increases clofarabine resistance in uterine serous carcinoma cells by a gain-of-function mechanism.

IF 6.6 2区 医学 Q1 Medicine
Cellular Oncology Pub Date : 2024-10-01 Epub Date: 2024-06-18 DOI:10.1007/s13402-024-00963-5
Michiel Remmerie, Rüveyda Dok, Zhigang Wang, Judit Domènech Omella, Sophie Alen, Célie Cokelaere, Lisa Lenaerts, Erwin Dreesen, Sandra Nuyts, Rita Derua, Veerle Janssens
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引用次数: 0

Abstract

Purpose: Uterine serous carcinoma (USC) is generally associated with poor prognosis due to a high recurrence rate and frequent treatment resistance; hence, there is a need for improved therapeutic strategies. Molecular analysis of USC identified several molecular markers, useful to improve current treatments or identify new druggable targets. PPP2R1A, encoding the Aα subunit of the tumor suppressive Ser/Thr phosphatase PP2A, is mutated in up to 40% of USCs. Here, we investigated the effect of the p.R183W PPP2R1A hotspot variant on treatment response to the nucleoside analogue clofarabine.

Methods and results: USC cells stably expressing p.R183W Aα showed increased resistance to clofarabine treatment in vitro and, corroborated by decreased clofarabine-induced apoptosis, G1 phase arrest, DNA-damage (γH2AX) and activation of ATM and Chk1/2 kinases. Phenotypic rescue by pharmacologic PP2A inhibition or dicer-substrate siRNA (dsiRNA)-mediated B56δ subunit knockdown supported a gain-of-function mechanism of Aα p.R183W, promoting dephosphorylation and inactivation of deoxycytidine kinase (dCK), the cellular enzyme responsible for the conversion of clofarabine into its bioactive form. Therapeutic assessment of related nucleoside analogues (gemcitabine, cladribine) revealed similar effects, but in a cell line-dependent manner. Expression of two other PPP2R1A USC mutants (p.P179R or p.S256F) did not affect clofarabine response in our cell models, arguing for mutant-specific effects on treatment outcome as well.

Conclusions: While our results call for PPP2R1A mutant and context-dependent effects upon clofarabine/nucleoside analogue monotherapy, combining clofarabine with a pharmacologic PP2A inhibitor proved synergistically in all tested conditions, highlighting a new generally applicable strategy to improve treatment outcome in USC.

Abstract Image

PPP2R1A 癌症热点突变体 p.R183W 通过功能增益机制增加了子宫浆液癌细胞对氯法拉滨的耐药性。
目的:子宫浆液性癌(USC)由于复发率高且经常出现耐药性,一般预后较差;因此,需要改进治疗策略。对子宫浆液性癌(USC)的分子分析发现了几个分子标记,有助于改善目前的治疗方法或确定新的药物靶点。编码肿瘤抑制性 Ser/Thr 磷酸酶 PP2A 的 Aα 亚基的 PPP2R1A 在高达 40% 的 USC 中发生突变。在此,我们研究了p.R183W PPP2R1A热点变异对核苷类似物氯法拉滨治疗反应的影响:体外稳定表达p.R183W Aα 的USC细胞对氯法拉滨治疗的耐药性增强,氯法拉滨诱导的细胞凋亡、G1期停滞、DNA损伤(γH2AX)以及ATM和Chk1/2激酶的活化减少也证实了这一点。药理 PP2A 抑制或 dicer-substrate siRNA(dsiRNA)介导的 B56δ 亚基敲除对表型的挽救支持了 Aα p.R183W 的功能增益机制,它促进了脱氧胞苷激酶(dCK)的去磷酸化和失活,而脱氧胞苷激酶是负责将氯法拉滨转化为其生物活性形式的细胞酶。对相关核苷类似物(吉西他滨、克拉利宾)的治疗评估显示了类似的效果,但这种效果依赖于细胞系。在我们的细胞模型中,另外两种 PPP2R1A USC 突变体(p.P179R 或 p.S256F)的表达并不影响氯法拉滨反应,这表明突变体对治疗结果也有特异性影响:我们的研究结果表明,PPP2R1A突变体和环境对氯法拉滨/核苷类似物单药治疗有依赖性影响,但在所有测试条件下,将氯法拉滨与药理PP2A抑制剂联合使用可产生协同作用,这将为改善USC的治疗效果提供一种普遍适用的新策略。
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来源期刊
Cellular Oncology
Cellular Oncology Biochemistry, Genetics and Molecular Biology-Cancer Research
CiteScore
10.40
自引率
1.50%
发文量
0
审稿时长
16 weeks
期刊介绍: The Official Journal of the International Society for Cellular Oncology Focuses on translational research Addresses the conversion of cell biology to clinical applications Cellular Oncology publishes scientific contributions from various biomedical and clinical disciplines involved in basic and translational cancer research on the cell and tissue level, technical and bioinformatics developments in this area, and clinical applications. This includes a variety of fields like genome technology, micro-arrays and other high-throughput techniques, genomic instability, SNP, DNA methylation, signaling pathways, DNA organization, (sub)microscopic imaging, proteomics, bioinformatics, functional effects of genomics, drug design and development, molecular diagnostics and targeted cancer therapies, genotype-phenotype interactions. A major goal is to translate the latest developments in these fields from the research laboratory into routine patient management. To this end Cellular Oncology forms a platform of scientific information exchange between molecular biologists and geneticists, technical developers, pathologists, (medical) oncologists and other clinicians involved in the management of cancer patients. In vitro studies are preferentially supported by validations in tumor tissue with clinicopathological associations.
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