Drápela Stanislav, Kvokačková Barbora, Slabáková Eva, Kotrbová Anna, Gömöryová Kristína, Fedr Radek, Kurfürstová Daniela, Eliáš Martin, Študent Vladimír, Lenčéšová Frederika, Ranjani Ganji Sri, Pospíchalová Vendula, Bryja Vítězslav, van Weerden Wytske M, Puhr Martin, Culig Zoran, Bouchal Jan, Souček Karel
Department of Cytokinetics, Institute of Biophysics of the Czech Academy of Sciences, Královopolská 135, Brno, 612 00, Czech Republic.
International Clinical Research Center, St. Anne's University Hospital in Brno, Brno, 602 00, Czech Republic.
Cell Oncol (Dordr). 2025 Feb;48(1):205-218. doi: 10.1007/s13402-024-00982-2. Epub 2024 Aug 20.
Docetaxel resistance is a significant obstacle in the treatment of prostate cancer (PCa), resulting in unfavorable patient prognoses. Intratumoral heterogeneity, often associated with epithelial-to-mesenchymal transition (EMT), has previously emerged as a phenomenon that facilitates adaptation to various stimuli, thus promoting cancer cell diversity and eventually resistance to chemotherapy, including docetaxel. Hence, understanding intratumoral heterogeneity is essential for better patient prognosis and the development of personalized treatment strategies.
To address this, we employed a high-throughput single-cell flow cytometry approach to identify a specific surface fingerprint associated with docetaxel-resistance in PCa cells and complemented it with proteomic analysis of extracellular vesicles. We further validated selected antigens using docetaxel-resistant patient-derived xenografts in vivo and probed primary PCa specimens to interrogate of their surface fingerprint.
Our approaches revealed a 6-molecule surface fingerprint linked to docetaxel resistance in primary PCa specimens. We observed consistent overexpression of CD95 (FAS/APO-1), and SSEA-4 surface antigens in both in vitro and in vivo docetaxel-resistant models, which was also observed in a cell subpopulation of primary PCa tumors exhibiting EMT features. Furthermore, CD95, along with the essential enzymes involved in SSEA-4 synthesis, ST3GAL1, and ST3GAL2, displayed a significant increase in patients with PCa undergoing docetaxel-based therapy, correlating with poor survival outcomes.
In summary, we demonstrate that the identified 6-molecule surface fingerprint associated with docetaxel resistance pre-exists in a subpopulation of primary PCa tumors before docetaxel treatment. Thus, this fingerprint warrants further validation as a promising predictive tool for docetaxel resistance in PCa patients prior to therapy initiation.
多西他赛耐药是前列腺癌（PCa）治疗中的一个重大障碍，导致患者预后不良。肿瘤内异质性通常与上皮-间质转化（EMT）相关，此前已成为一种促进对各种刺激适应的现象，从而促进癌细胞多样性并最终导致对包括多西他赛在内的化疗产生耐药性。因此，了解肿瘤内异质性对于改善患者预后和制定个性化治疗策略至关重要。
为了解决这个问题，我们采用了高通量单细胞流式细胞术方法来识别与PCa细胞中多西他赛耐药相关的特定表面特征，并通过细胞外囊泡的蛋白质组学分析对其进行补充。我们使用多西他赛耐药患者来源的异种移植物在体内进一步验证了选定的抗原，并对原发性PCa标本进行检测以探究其表面特征。
我们的方法揭示了原发性PCa标本中与多西他赛耐药相关的6分子表面特征。我们在体外和体内多西他赛耐药模型中均观察到CD95（FAS/APO-1）和SSEA-4表面抗原的一致过表达，在具有EMT特征的原发性PCa肿瘤细胞亚群中也观察到了这一现象。此外，在接受基于多西他赛治疗的PCa患者中，CD95以及参与SSEA-4合成的关键酶ST3GAL1和ST3GAL2显著增加，这与不良生存结果相关。
总之，我们证明在多西他赛治疗之前，原发性PCa肿瘤亚群中预先存在与多西他赛耐药相关的已识别6分子表面特征。因此，这一特征作为PCa患者治疗开始前多西他赛耐药的一种有前景的预测工具，值得进一步验证。
