Monzer Alissar, Ghamlouche Fatima, Wakimian Kevork, Ballout Farah, Al Bitar Samar, Yehya Amani, Kanso Mariam, Saheb Nour, Tawil Ayman, Doughan Samer, Hussein Maher, Mukherji Deborah, Faraj Walid, Allen Joshua E, Prabhu Varun V, Abou-Antoun Tamara, Gali-Muhtasib Hala, Abou-Kheir Wassim
Department of Anatomy, Cell Biology and Physiological Sciences, Faculty of Medicine, American University of Beirut, Beirut, 1107-2020, Lebanon.
Department of Surgery, American University of Beirut Medical Center, Beirut, 1107-2020, Lebanon.
Pharmacol Rep. 2025 Feb;77(1):229-246. doi: 10.1007/s43440-024-00676-4. Epub 2024 Nov 18.
Colorectal cancer (CRC) remains one of the most frequently diagnosed and life-threatening malignancies worldwide. CRC's high recurrence rates and drug resistance have been correlated with a subpopulation of dormant slowly dividing cells termed CRC stem cells (CCSCs). Consequently, there is a pressing need to identify novel therapeutics that can effectively and specifically target CCSCs. Imipridones are promising structurally related anticancer molecules that showed efficacy in several solid and hematological preclinical models and phase I/II/III clinical trials. This study mainly aimed to assess the potential anticancer effects of ONC206, an imipridone derivative, on CRC three-dimensional in vitro culture systems using HCT116 and HT29 cells. Importantly, the study aimed at using CRC patient-derived organoids (PDOs) to test the potential therapeutic effect of ONC206.
Two-dimensional cell proliferation, viability, migration, and invasion assays were used to assess the effects of ONC206 on two colorectal cancer cell lines, HCT116 and HT29, in vitro. Immunofluorescence imaging, flow cytometry, and western blot analysis were also performed to investigate the mechanism of action of this drug. Sphere formation assay and CRC PDOs were employed to evaluate the effect of ONC206 on CRC cells in a 3D setting and specifically its potency in targeting the CRC stem/progenitor subpopulation of cells.
Our results showed that ONC206 was more potent than its parental molecule ONC201 in inhibiting the proliferation and viability of HCT116 and HT29 cells. Moreover, ONC206 significantly reduced the migration and invasion indices of CRC cells. These effects were accompanied by an increase in reactive oxygen species (ROS) production, sub-G1 phase accumulation, and apoptosis in HCT116 and HT29 cells. Furthermore, ONC206 significantly inhibited the 3D colonospheres growth and self-renewal ability of CCSCs more potently than ONC201, which was associated with a decrease in the expression of CSC-related markers. Lastly, ONC206 significantly reduced the growth of organoids derived from CRC patients.
Collectively, our findings demonstrate that ONC206 is an effective anticancer molecule capable of targeting CCSCs, which may represent a novel therapeutic strategy that can overcome CRC resistance and recurrence.
结直肠癌（CRC）仍然是全球最常被诊断出且危及生命的恶性肿瘤之一。CRC的高复发率和耐药性与一群称为CRC干细胞（CCSCs）的休眠缓慢分裂细胞亚群有关。因此，迫切需要鉴定能够有效且特异性靶向CCSCs的新型疗法。咪吡酮类是有前景的结构相关抗癌分子，在几种实体瘤和血液学临床前模型以及I/II/III期临床试验中显示出疗效。本研究主要旨在评估咪吡酮衍生物ONC206对使用HCT116和HT29细胞的CRC三维体外培养系统的潜在抗癌作用。重要的是，该研究旨在使用CRC患者来源的类器官（PDOs）来测试ONC206的潜在治疗效果。
使用二维细胞增殖、活力、迁移和侵袭试验来评估ONC206对两种结肠癌细胞系HCT116和HT29的体外作用。还进行了免疫荧光成像、流式细胞术和蛋白质印迹分析以研究该药物的作用机制。采用成球试验和CRC PDOs来评估ONC206在三维环境中对CRC细胞的作用，特别是其靶向CRC干细胞/祖细胞亚群细胞的效力。
我们的结果表明，ONC206在抑制HCT116和HT29细胞的增殖和活力方面比其母体分子ONC201更有效。此外，ONC206显著降低了CRC细胞的迁移和侵袭指数。这些作用伴随着HCT116和HT29细胞中活性氧（ROS）产生增加、亚G1期积累和细胞凋亡。此外，ONC206比ONC201更有效地显著抑制CCSCs的三维结肠球生长和自我更新能力，这与CSC相关标志物表达的降低有关。最后，ONC206显著降低了CRC患者来源的类器官的生长。
总体而言，我们的研究结果表明ONC206是一种能够靶向CCSCs的有效抗癌分子，这可能代表一种可以克服CRC耐药性和复发的新型治疗策略。
