The component of the m(6)A writer complex VIRMA is implicated in aggressive tumor phenotype, DNA damage response and cisplatin resistance in germ cell tumors.
The component of the m(6)A writer complex VIRMA is implicated in aggressive tumor phenotype, DNA damage response and cisplatin resistance in germ cell tumors.
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DOI：
10.1186/s13046-021-02072-9
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发表时间：
2021-08-25
期刊：
影响因子：
--
通讯作者：
Jerónimo C
中科院分区：
文献类型：
--
作者：
Miranda-Gonçalves V;Lobo J;Guimarães-Teixeira C;Barros-Silva D;Guimarães R;Cantante M;Braga I;Maurício J;Oing C;Honecker F;Nettersheim D;Looijenga LHJ;Henrique R;Jerónimo C
生殖细胞肿瘤（GCT）是发育性癌症，与胚胎发生和生殖细胞发育密切相关。最近和不断扩大的RNA修饰领域越来越多地涉及这些分子事件，以及肿瘤进展和对治疗的抗性，但在GCT中仍然很少探索。在这项工作中，作为我们最近在TGCT组织样本中对这一主题进行的研究的后续工作，我们的目标是研究N6-甲基腺苷（m6 A）的作用，这是mRNA中最丰富的此类修饰，在体外和体内模型中代表此类肿瘤。使用了四种代表GCT的细胞系（三种睾丸细胞系和一种纵隔细胞系），包括一种等基因顺铂耐药亚系。建立CRISPR/Cas9介导的VIRMA敲低，并使用绒毛尿囊膜测定来研究其体内表型效应。我们证明了不同的m6 A作家，读者和橡皮擦在GCT细胞系的主要类别，这些肿瘤，腺瘤和非腺瘤的代表性的差异表达，我们证明了分化与全反式维甲酸治疗后发生的变化。我们还显示了对顺铂治疗敏感和耐药的细胞之间的差异表达，暗示这些参与者获得顺铂耐药表型。VIRMA的敲除导致剩余的甲基转移酶复合物的破坏和m6 A丰度的降低，以及总体上降低的肿瘤侵袭性（细胞活力、肿瘤细胞增殖、迁移和侵袭降低）和对顺铂治疗的敏感性增加，这在体外和体内均得到证实。VIRMA敲除后对顺铂的反应增强与DNA损伤显著增加（γ H2 AX和GADD 45 B水平较高）以及XLF和MRE 11下调相关。VIRMA在GCT中具有致癌作用，证实了我们先前基于组织的研究，并通过干扰DNA修复进一步参与顺铂的反应。这些数据有助于我们更好地了解GCT中顺铂耐药性的出现，并支持最近针对m6 A写入复合物元素的治疗尝试。在线版本包含补充材料，可通过10.1186/s13046-021-02072-9获得。
Germ cell tumors (GCTs) are developmental cancers, tightly linked to embryogenesis and germ cell development. The recent and expanding field of RNA modifications is being increasingly implicated in such molecular events, as well as in tumor progression and resistance to therapy, but still rarely explored in GCTs. In this work, and as a follow-up of our recent study on this topic in TGCT tissue samples, we aim to investigate the role of N6-methyladenosine (m6A), the most abundant of such modifications in mRNA, in in vitro and in vivo models representative of such tumors. Four cell lines representative of GCTs (three testicular and one mediastinal), including an isogenic cisplatin resistant subline, were used. CRISPR/Cas9-mediated knockdown of VIRMA was established and the chorioallantoic membrane assay was used to study its phenotypic effect in vivo. We demonstrated the differential expression of the various m6A writers, readers and erasers in GCT cell lines representative of the major classes of these tumors, seminomas and non-seminomas, and we evidenced changes occurring upon differentiation with all-trans retinoic acid treatment. We showed differential expression also among cells sensitive and resistant to cisplatin treatment, implicating these players in acquisition of cisplatin resistant phenotype. Knockdown of VIRMA led to disruption of the remaining methyltransferase complex and decrease in m6A abundance, as well as overall reduced tumor aggressiveness (with decreased cell viability, tumor cell proliferation, migration, and invasion) and increased sensitivity to cisplatin treatment, both in vitro and confirmed in vivo. Enhanced response to cisplatin after VIRMA knockdown was related to significant increase in DNA damage (with higher γH2AX and GADD45B levels) and downregulation of XLF and MRE11. VIRMA has an oncogenic role in GCTs confirming our previous tissue-based study and is further involved in response to cisplatin by interfering with DNA repair. These data contribute to our better understanding of the emergence of cisplatin resistance in GCTs and support recent attempts to therapeutically target elements of the m6A writer complex. The online version contains supplementary material available at 10.1186/s13046-021-02072-9.
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影响因子：
2
作者：
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作者：
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影响因子：
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