在晚期临床前3D黑色素瘤模型中进行高通量药物筛选,确定了nras突变黑色素瘤的潜在一线治疗方法。

IF 12.8 1区 医学 Q1 ONCOLOGY
Cristian Angeli, Demetra Philippidou, Eliane Klein, Christiane Margue, Sagarika Ghosh, Maria Lorena Cordero Maldonado, Natascia Tiso, Giovanni Risato, Fizza Irfan, Bruno Santos, Meritxell Cutrona, Joanna Patrycja Wroblewska, Stephanie Kreis
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引用次数: 0

摘要

背景:尽管靶向(BRAFi + MEKi)和免疫(抗pd1 /PD-L1、抗ctla4和抗lag3)疗法取得了重大进展,但NRASmut黑色素瘤的治疗选择仍然有限。目前,NRASmut患者依赖免疫检查点抑制剂、经典化疗和标签外MEK抑制剂,超过50%的患者经历快速的疾病进展。开发有效靶向治疗的关键挑战之一是缺乏准确概括肿瘤微环境(TME)和携带NRAS突变的黑色素瘤细胞内在抗性的临床前模型。方法:为了解决这个问题,我们对3D NRASmut黑色素瘤球体进行了1300多种化合物的高通量筛选(HTS)。采用多步骤分析确定命中点,并通过药物反应曲线(DRC)分析进一步检验命中点。大多数有希望的化合物通过单培养和共培养3D体外模型进一步验证,模拟黑色素瘤的三个主要转移部位,如皮肤/真皮,肺和肝脏,利用球体和水凝胶系统。最后,使用斑马鱼异种移植模型进行验证,以便更精细和准确地评估药物反应。结果:NRASmut黑色素瘤球体的高通量药物筛选鉴定出17种候选化合物,随后通过DRC分析验证。在最有希望的药物中,柔红霉素HCl (DH)和帕莫酸Pyrvinium Pamoate (PP)被选中进行进一步的研究,在先进的3D共培养系统和斑马鱼异种移植模型中显示出强大的抗黑色素瘤活性。值得注意的是,与未经批准的MEK抑制剂Trametinib相比,PP显示出更高的细胞毒性,对AKT和患者源性转移性黑色素瘤细胞系的侵袭行为具有抑制作用。此外,与曲美替尼联合治疗导致细胞增殖和活力的累加效应。重要的是,这两种化合物在耐曲美替尼(MEK抑制剂)的NRASmut和BRAFwt/NRASwt黑色素瘤细胞系中显示出相似的疗效。结论:本研究利用包含关键TME元素的先进3D黑色素瘤模型和斑马鱼异种移植模型,强调了盐酸柔红霉素和帕莫酸吡啶盐作为NRASmut黑色素瘤的新型一线疗法的潜力,对meki耐药细胞也有显著的影响。这些发现支持了药物再利用策略,并强调了生理相关的临床前模型在确定有效疗法中的重要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High-throughput drug screening in advanced pre-clinical 3D melanoma models identifies potential first-line therapies for NRAS-mutated melanoma.

Background: Despite significant advances in targeted (BRAFi + MEKi) and immune (anti-PD1/PD-L1, anti-CTLA4, and anti-LAG3) therapies, treatment options for NRASmut melanoma remain limited. Currently, NRASmut patients rely on immune checkpoint inhibitors, classical chemotherapy, and off-label MEK inhibitors, with over 50% experiencing rapid disease progression. One of the key challenges in developing effective targeted therapies is the lack of preclinical models that accurately recapitulate the tumor microenvironment (TME) and the intrinsic resistance of melanoma cells bearing NRAS mutations.

Methods: To address this, we performed high-throughput screening (HTS) of over 1,300 compounds in 3D NRASmut melanoma spheroids. A multi-step analysis was performed to identify hits, which were further tested by performing drug-response curve (DRC) analysis. Most promising compounds were further validated using mono- and co-culture 3D in vitro models that mimic three main metastatic sites in melanoma, such as skin/dermal, lung, and liver, utilizing spheroid and hydrogel systems. Ultimately, validation was conducted using zebrafish xenograft models to enable a more refined and accurate assessment of drug response.

Results: High-throughput drug screening of NRASmut melanoma spheroids identified 17 candidate compounds, which were subsequently validated through DRC analyses. Among the most promising drugs, Daunorubicin HCl (DH) and Pyrvinium Pamoate (PP) were selected for further investigation, demonstrating potent anti-melanoma activity in advanced 3D co-culture systems and zebrafish xenograft models. Notably, PP demonstrated higher cytotoxicity compared to Trametinib, the off-label MEK inhibitor, with an inhibitory effect on AKT and invasive behavior in the patient-derived metastatic melanoma cell lines. Additionally, combinatorial treatment with Trametinib resulted in additive effects on cell proliferation and viability. Importantly, both compounds showed similar efficacy in NRASmut and BRAFwt/NRASwt melanoma cell lines that were resistant to Trametinib (MEK inhibitor).

Conclusions: Using advanced 3D melanoma models that incorporate key TME elements and zebrafish xenograft models, this study highlights the potential of Daunorubicin HCl and Pyrvinium Pamoate as novel first-line therapies for NRASmut melanoma, with a noteworthy effect also on MEKi-resistant cells. These findings support drug repurposing strategies and underscore the importance of physiologically relevant preclinical models in identifying effective therapies.

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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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