WDR3 undergoes phase separation to mediate the therapeutic mechanism of Nilotinib against osteosarcoma.

IF 12.8 1区 医学 Q1 ONCOLOGY
Minglei Li, Nan Li, Yuying Fan, Zhan Zhang, Long Zhou, Yifan Yu, Man Ni, Mingzi Tan, WanJie Huang, Tong Zhu
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引用次数: 0

Abstract

Background: Osteosarcoma is highly invasive with a poor prognosis. The phenomenon of liquid-liquid phase separation (LLPS) can promote the formation of biomolecules and participate in the tumor regulation mechanism. Therefore, mining prognostic markers related to LLPS could allow patients to benefit from targeted therapies.

Method: Microarray analysis was performed to identify LLPS-related biomarkers, followed by the validation of binding interactions between genes and drugs via molecular docking analysis. Functions of key genes were investigated in U2-OS cells and xenograft mice. LLPS of WDR3 were observed by the droplet formation assay and fluorescence recovery after photobleaching. The intrinsically disordered region (IDR) of WDR3 was mutated to disrupt LLPS, which was then rescued by the fusion of hnRNAP1 IDR. Therapeutic mechanism of Nilotinib mediated by LLPS was explored in vitro and in vivo.

Results: Five LLPS-related biomarkers were screened by bioinformatics analyses to predict the osteosarcoma prognosis. These prognostic genes were significantly associated with the immune cell infiltration, tumor immune escape and drug sensitivity. Among them, WDR3 was a prognostic risk factor for osteosarcoma and stably bound to Nilotinib in the molecular docking model. In transfected U2-OS cells and xenograft mice, the downregulation of WDR3 significantly inhibited the malignant progression of osteosarcoma. More importantly, WDR3 could form droplets in U2-OS cells and restore the fluorescence intensity of WDR3 condensates with liquid-like behavior after photobleaching. The mutation in IDR impaired the phase separation ability of WDR3, whereas the fusion with hnRNAP1 IDR rescued the phase separation abnormality caused by WDR3 mutation. Moreover, the treatment with Nilotinib improved the progression of osteosarcoma in vivo and in vitro, while inhibiting the production of WDR3 phase-separated condensates.

Conclusion: WDR3 phase separation involves in the therapeutic mechanism of Nilotinib against osteosarcoma, and thus may serve as a potent biomarker to ameliorate adverse events following osteosarcoma treatment.

WDR3通过相分离介导尼洛替尼对骨肉瘤的治疗机制。
背景:骨肉瘤侵袭性强,预后差。液-液相分离(LLPS)现象可促进生物分子的形成,参与肿瘤调控机制。因此,挖掘与LLPS相关的预后标志物可以使患者从靶向治疗中获益。方法:通过微阵列分析鉴定llps相关生物标志物,通过分子对接分析验证基因与药物之间的结合相互作用。在U2-OS细胞和异种移植小鼠中研究了关键基因的功能。采用液滴形成法和光漂白后荧光恢复法观察WDR3的LLPS。WDR3的内在无序区(IDR)突变破坏LLPS,然后通过hnRNAP1 IDR的融合恢复LLPS。探讨了LLPS介导尼洛替尼的体内外治疗机制。结果:通过生物信息学分析筛选了5个与llps相关的生物标志物,预测骨肉瘤的预后。这些预后基因与免疫细胞浸润、肿瘤免疫逃逸和药物敏感性显著相关。其中,WDR3是骨肉瘤的预后危险因素,在分子对接模型中与尼洛替尼稳定结合。在转染的U2-OS细胞和异种移植小鼠中,下调WDR3可显著抑制骨肉瘤的恶性进展。更重要的是,WDR3可以在U2-OS细胞中形成液滴,使光漂白后WDR3凝聚物的荧光强度恢复为液体状。IDR的突变削弱了WDR3的相分离能力,而与hnRNAP1 IDR的融合恢复了WDR3突变引起的相分离异常。此外,尼洛替尼治疗改善了体内和体外骨肉瘤的进展,同时抑制了WDR3相分离凝聚物的产生。结论:WDR3相分离参与了尼洛替尼治疗骨肉瘤的机制,因此可能作为一种有效的生物标志物来改善骨肉瘤治疗后的不良事件。
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来源期刊
CiteScore
18.20
自引率
1.80%
发文量
333
审稿时长
1 months
期刊介绍: The Journal of Experimental & Clinical Cancer Research is an esteemed peer-reviewed publication that focuses on cancer research, encompassing everything from fundamental discoveries to practical applications. We welcome submissions that showcase groundbreaking advancements in the field of cancer research, especially those that bridge the gap between laboratory findings and clinical implementation. Our goal is to foster a deeper understanding of cancer, improve prevention and detection strategies, facilitate accurate diagnosis, and enhance treatment options. We are particularly interested in manuscripts that shed light on the mechanisms behind the development and progression of cancer, including metastasis. Additionally, we encourage submissions that explore molecular alterations or biomarkers that can help predict the efficacy of different treatments or identify drug resistance. Translational research related to targeted therapies, personalized medicine, tumor immunotherapy, and innovative approaches applicable to clinical investigations are also of great interest to us. We provide a platform for the dissemination of large-scale molecular characterizations of human tumors and encourage researchers to share their insights, discoveries, and methodologies with the wider scientific community. By publishing high-quality research articles, reviews, and commentaries, the Journal of Experimental & Clinical Cancer Research strives to contribute to the continuous improvement of cancer care and make a meaningful impact on patients' lives.
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