Use of non-small cell lung cancer multicellular tumor spheroids to study the impact of chemotherapy

IF 4.7 2区 医学 Q1 RESPIRATORY SYSTEM Respiratory Research Pub Date : 2024-04-05 DOI:10.1186/s12931-024-02791-5
Pauline Hulo, Sophie Deshayes, Judith Fresquet, Anne-Laure Chéné, Stéphanie Blandin, Nicolas Boisgerault, Jean-François Fonteneau, Lucas Treps, Marc G Denis, Jaafar Bennouna, Delphine Fradin, Elvire Pons-Tostivint, Christophe Blanquart
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Abstract

Lung cancers represent the main cause of cancer related-death worldwide. Recently, immunotherapy alone or in combination with chemotherapy has deeply impacted the therapeutic care leading to an improved overall survival. However, relapse will finally occur, with no efficient second line treatment so far. New therapies development based on the comprehension of resistance mechanisms is necessary. However, the difficulties to obtain tumor samples before and after first line treatment hamper to clearly understand the consequence of these molecules on tumor cells and also to identify adapted second line therapies. To overcome this difficulty, we developed multicellular tumor spheroids (MCTS) using characterized Non-Small Cell Lung Cancer (NSCLC) cell lines, monocytes from healthy donors and fibroblasts. MCTS were treated with carboplatin-paclitaxel or -gemcitabine combinations according to clinical administration schedules. The treatments impact was studied using cell viability assay, histological analyses, 3’RNA sequencing, real-time PCR, flow cytometry and confocal microscopy. We showed that treatments induced a decrease in cell viability and strong modifications in the transcriptomic profile notably at the level of pathways involved in DNA damage repair and cell cycle. Interestingly, we also observed a modification of genes expression considered as hallmarks of response to immune check point inhibitors and immunogenicity, particularly an increase in CD274 gene expression, coding for PD-L1. This result was validated at the protein level and shown to be restricted to tumor cells on MCTS containing fibroblasts and macrophages. This increase was also observed in an additional cell line, expressing low basal CD274 level. This study shows that MCTS are interesting models to study the impact of first line therapies using conditions close to clinical practice and also to identify more adapted second line or concomitant therapies for lung cancer treatment.
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利用非小细胞肺癌多细胞肿瘤球研究化疗的影响
肺癌是全球癌症相关死亡的主要原因。最近,免疫疗法单独使用或与化疗联合使用对治疗护理产生了深远影响,从而提高了总生存率。然而,癌症终究会复发,迄今为止还没有有效的二线治疗方法。有必要在了解抗药性机制的基础上开发新的疗法。然而,由于在一线治疗前后难以获得肿瘤样本,因此无法清楚地了解这些分子对肿瘤细胞的影响,也无法确定适合的二线疗法。为了克服这一困难,我们利用特征化的非小细胞肺癌(NSCLC)细胞系、健康供体的单核细胞和成纤维细胞开发了多细胞肿瘤球(MCTS)。根据临床用药计划,用卡铂-紫杉醇或吉西他滨组合治疗 MCTS。使用细胞活力测定、组织学分析、3'RNA 测序、实时 PCR、流式细胞术和共聚焦显微镜研究了治疗的影响。我们发现,治疗会导致细胞存活率下降,并使转录组谱发生强烈变化,特别是在 DNA 损伤修复和细胞周期相关通路的水平上。有趣的是,我们还观察到作为免疫检查点抑制剂反应和免疫原性标志的基因表达发生了改变,尤其是编码 PD-L1 的 CD274 基因表达增加。这一结果在蛋白质水平上得到了验证,并显示仅限于含有成纤维细胞和巨噬细胞的 MCTS 上的肿瘤细胞。在另一种基础 CD274 表达水平较低的细胞系中也观察到了这种增加。这项研究表明,MCTS 是一种有趣的模型,可用于在接近临床实践的条件下研究一线疗法的影响,也可用于确定更适合肺癌治疗的二线疗法或伴随疗法。
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来源期刊
Respiratory Research
Respiratory Research 医学-呼吸系统
自引率
1.70%
发文量
314
期刊介绍: Respiratory Research publishes high-quality clinical and basic research, review and commentary articles on all aspects of respiratory medicine and related diseases. As the leading fully open access journal in the field, Respiratory Research provides an essential resource for pulmonologists, allergists, immunologists and other physicians, researchers, healthcare workers and medical students with worldwide dissemination of articles resulting in high visibility and generating international discussion. Topics of specific interest include asthma, chronic obstructive pulmonary disease, cystic fibrosis, genetics, infectious diseases, interstitial lung diseases, lung development, lung tumors, occupational and environmental factors, pulmonary circulation, pulmonary pharmacology and therapeutics, respiratory immunology, respiratory physiology, and sleep-related respiratory problems.
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