破坏 YAP1 介导的谷氨酰胺代谢可诱导骨肉瘤中的合成致死率,同时抑制 ODC1。

IF 4.9 2区 医学 Q2 CELL BIOLOGY
Cellular Oncology Pub Date : 2024-10-01 Epub Date: 2024-08-08 DOI:10.1007/s13402-024-00967-1
Hongsheng Wang, Yining Tao, Jing Han, Jiakang Shen, Haoran Mu, Zhuoying Wang, Jinzeng Wang, Xinmeng Jin, Qi Zhang, Yuqin Yang, Jun Lin, Mengxiong Sun, Xiaojun Ma, Ling Ren, Amy K LeBlanc, Jing Xu, Yingqi Hua, Wei Sun
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引用次数: 0

摘要

目的:骨肉瘤是一种高度恶性的原发性骨肿瘤,主要影响青少年,经常对初始化疗产生耐药性,导致转移和治疗选择有限。我们的研究旨在发现治疗转移性和复发性骨肉瘤的新靶点:在这项研究中,我们证明了调节 YAP1 调控的谷氨酰胺代谢途径可增强 OS 对 DFMO 的反应。我们首先利用单细胞转录组数据来衡量MTAP缺失的OS患者体内多胺代谢的激活水平。复发性和非复发性患者组织的转录组测序数据进一步证实了这一点,证实了进行性OS中多胺代谢的激活。通过高通量药物筛选,我们发现 YAP1 抑制剂 CIL56 是与 DFMO 联合治疗策略的理想候选药物。在体内,我们利用 PDX 和 CDX 模型来验证这种药物组合的疗效。在体外,我们进行了 Western 印迹分析、qPCR 分析、免疫荧光染色和 PuMA 实验,以监测分子表达、分布和肿瘤转移能力的变化。我们采用 CCK-8 和集落形成试验来评估实验组细胞的增殖能力。我们使用流式细胞仪和活性氧探针观察细胞内 ROS 和谷氨酰胺代谢的变化。最后,我们将 RNA-seq 与代谢组学结合起来,确定了经 DFMO 和 CIL56 组合处理的 OS 细胞的代谢变化。这使我们能够干预和验证 YAP1 介导的谷氨酰胺代谢途径在 DFMO 抗性中的作用:结果:通过单细胞RNA-seq数据分析,我们确定了多胺代谢显著上调的晚期OS细胞亚群。复发性和非复发性 OS 组织的转录组分析进一步证实了这种上调。高通量药物筛选显示,DFMO和CIL56的组合策略很有前景。DFMO 治疗可抑制 OS 细胞中 YAP1 蛋白的磷酸化,促进核进入并启动 YAP1 介导的谷氨酰胺代谢途径。这降低了细胞内的 ROS 水平,抵消了 DFMO 的抗癌作用。将 DFMO 与 YAP1 抑制剂 CIL56 或谷氨酰胺酶抑制剂 CB-839 结合使用,可在体内和体外扩大 DFMO 的疗效。这凸显了靶向YAP1介导的谷氨酰胺代谢途径以提高DFMO疗效的巨大潜力:结论:我们的研究结果阐明了 YAP1 介导的谷氨酰胺代谢是继抑制多胺代谢之后对 DFMO 起关键作用的旁路机制。我们的研究为 DFMO 在转移性和复发性骨肉瘤的 "一对二冲剂 "疗法中的潜在作用提供了有价值的见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Disrupting YAP1-mediated glutamine metabolism induces synthetic lethality alongside ODC1 inhibition in osteosarcoma.

Disrupting YAP1-mediated glutamine metabolism induces synthetic lethality alongside ODC1 inhibition in osteosarcoma.

Purpose: Osteosarcoma, a highly malignant primary bone tumor primarily affecting adolescents, frequently develops resistance to initial chemotherapy, leading to metastasis and limited treatment options. Our study aims to uncover novel therapeutic targets for metastatic and recurrent osteosarcoma.

Methods: In this study, we proved the potential of modulating the YAP1-regulated glutamine metabolic pathway to augment the response of OS to DFMO. We initially employed single-cell transcriptomic data to gauge the activation level of polyamine metabolism in MTAP-deleted OS patients. This was further substantiated by transcriptome sequencing data from recurrent and non-recurrent patient tissues, confirming the activation of polyamine metabolism in progressive OS. Through high-throughput drug screening, we pinpointed CIL56, a YAP1 inhibitor, as a promising candidate for a combined therapeutic strategy with DFMO. In vivo, we utilized PDX and CDX models to validate the therapeutic efficacy of this drug combination. In vitro, we conducted western blot analysis, qPCR analysis, immunofluorescence staining, and PuMA experiments to monitor alterations in molecular expression, distribution, and tumor metastasis capability. We employed CCK-8 and colony formation assays to assess the proliferative capacity of cells in the experimental group. We used flow cytometry and reactive oxygen probes to observe changes in ROS and glutamine metabolism within the cells. Finally, we applied RNA-seq in tandem with metabolomics to identify metabolic alterations in OS cells treated with a DFMO and CIL56 combination. This enabled us to intervene and validate the role of the YAP1-mediated glutamine metabolic pathway in DFMO resistance.

Results: Through single-cell RNA-seq data analysis, we pinpointed a subset of late-stage OS cells with significantly upregulated polyamine metabolism. This upregulation was further substantiated by transcriptomic profiling of recurrent and non-recurrent OS tissues. High-throughput drug screening revealed a promising combination strategy involving DFMO and CIL56. DFMO treatment curbs the phosphorylation of YAP1 protein in OS cells, promoting nuclear entry and initiating the YAP1-mediated glutamine metabolic pathway. This reduces intracellular ROS levels, countering DFMO's anticancer effect. The therapeutic efficacy of DFMO can be amplified both in vivo and in vitro by combining it with the YAP1 inhibitor CIL56 or the glutaminase inhibitor CB-839. This underscores the significant potential of targeting the YAP1-mediated glutamine metabolic pathway to enhance efficacy of DFMO.

Conclusion: Our findings elucidate YAP1-mediated glutamine metabolism as a crucial bypass mechanism against DFMO, following the inhibition of polyamine metabolism. Our study provides valuable insights into the potential role of DFMO in an "One-two Punch" therapy of metastatic and recurrent osteosarcoma.

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来源期刊
Cellular Oncology
Cellular Oncology ONCOLOGY-CELL BIOLOGY
CiteScore
10.30
自引率
1.50%
发文量
86
审稿时长
12 months
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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