Richard Mathilde, Moreau Rosalie, Croyal Mikaël, Mathiot Laurent, Frénel Jean-Sébastien, Campone Mario, Dupont Aurélien, Gavard Julie, André-Grégoire Gwennan, Guével Laëtitia
Team SOAP, Centre de Recherche en Cancérologie et Immunologie Intégrée Nantes-Angers (CRCI2NA), Inserm CNRS, Nantes Université Nantes France.
Équipe Labellisée Ligue Contre le Cancer Paris France.
J Extracell Biol. 2024 Dec 17;3(12):e70013. doi: 10.1002/jex2.70013. eCollection 2024 Dec.
Extracellular vesicles (EVs) are cell-derived small membrane structures that transport various molecules. They have emerged as potential circulating biomarkers for monitoring responses to cancer therapies. This study aimed to comprehensively characterize plasma-carried EVs in hormone receptor-positive (HR) metastatic breast cancer (MBC) patients treated with first-line CDK4/6 inhibitors (iCDK4/6) combined with endocrine therapy. MBC patients were classified into three groups based on their response to therapy: resistant, intermediate or sensitive. In a prospective cohort, we monitored the concentration of circulating EVs, analyzed their lipid signature and correlated these factors with treatment response. To facilitate the translation of EV research to clinical practice, we established a three-step procedure: (1) EVs were isolated from plasma using semi-automatized size exclusion chromatography (SEC); (2) EV concentration, termed vesiclemia, was determined by drop counting via interferometric light microscopy (ILM); and (3) EV lipid composition was analyzed by mass spectrometry. ILM-based vesiclemia values were highly fluctuating upon iCDK4/6 treatment, while early increase associated with accelerated progression. Of note, vesiclemia remained a steady parameter over a 1-year period in age-matched healthy women. Additionally, analysis of the EV cargo unveiled a distinct sphingolipid profile, characterized by increased levels of ceramides and sphingomyelins in resistant patients within the first 2 months of treatment. Based on 16 sphingolipid species, sensitive and resistant patients were correctly classified with an overall accuracy of 82%. This specific sphingolipid pattern was exclusively discernible within EVs, and not in plasma, highlighting the significance of EVs in the early prediction of individual responses to iCDK4/6 and disease progression. Overall, this study provides insights of the longitudinal characterization of plasma-borne EVs in both a healthy group and HR MBC patients under iCDK4/6 therapies. Combined vesiclemia and EV sphingolipid profile emphasize the promising potential of EVs as non-invasive biomarkers for monitoring early treatment response.
细胞外囊泡（EVs）是细胞衍生的小膜结构，可运输各种分子。它们已成为监测癌症治疗反应的潜在循环生物标志物。本研究旨在全面表征接受一线CDK4/6抑制剂（iCDK4/6）联合内分泌治疗的激素受体阳性（HR）转移性乳腺癌（MBC）患者血浆携带的细胞外囊泡。MBC患者根据其治疗反应分为三组：耐药、中度反应或敏感。在一个前瞻性队列中，我们监测了循环细胞外囊泡的浓度，分析了它们的脂质特征，并将这些因素与治疗反应相关联。为了促进细胞外囊泡研究向临床实践的转化，我们建立了一个三步程序：（1）使用半自动尺寸排阻色谱法（SEC）从血浆中分离细胞外囊泡；（2）通过干涉光显微镜（ILM）计数液滴来确定细胞外囊泡浓度，即囊泡血症；（3）通过质谱分析细胞外囊泡脂质组成。基于ILM的囊泡血症值在iCDK4/6治疗期间波动很大，而早期升高与疾病进展加速相关。值得注意的是，在年龄匹配的健康女性中，囊泡血症在1年期间保持稳定参数。此外，对细胞外囊泡货物的分析揭示了一种独特的鞘脂谱，其特征是在治疗的前2个月内，耐药患者的神经酰胺和鞘磷脂水平升高。基于16种鞘脂种类，敏感和耐药患者被正确分类，总体准确率为82%。这种特定的鞘脂模式仅在细胞外囊泡中可辨别，而在血浆中不可见，突出了细胞外囊泡在早期预测个体对iCDK4/6的反应和疾病进展中的重要性。总体而言，本研究提供了在健康组和接受iCDK4/6治疗的HR MBC患者中血浆携带的细胞外囊泡纵向特征的见解。联合囊泡血症和细胞外囊泡鞘脂谱强调了细胞外囊泡作为监测早期治疗反应的非侵入性生物标志物的潜在前景。
