PO.ET09.06 · 实验与分子治疗
使用 HMCD-青蒿素偶联物对转移性去势抵抗性前列腺癌进行亚细胞靶向治疗
Subcellular targeted therapy using HMCD-artemisinin conjugates for metastatic castration-resistant prostate cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景:对雄激素剥夺治疗的耐药导致转移性去势抵抗性前列腺癌(mCRPC),这是一个治疗选择有限的致命阶段。青蒿素(ART)衍生物通过活性氧(ROS)生成和线粒体破坏发挥抗肿瘤活性,但缺乏肿瘤选择性。我们的实验室鉴定出一种近红外(NIR)七甲川碳菁染料(HMCD),它通过在缺氧条件下上调的有机阴离子转运多肽(OATP)优先在肿瘤中蓄积,从而实现肿瘤特异性递送。
方法:为实现亚细胞靶向化疗,合成了四种带有不同连接子的 HMCD-ART 偶联物:HMCD-ART1(青蒿琥酯连接子)、HMCD-ART2(氨基甲酸酯连接子)、HMCD-ART3(醚连接子)和 HMCD-ART4(酯连接子)。在雄激素非依赖性前列腺癌模型(22Rv1、PC-3、C4-2B 及耐药衍生株)中对它们进行了评估。在体外分析了细胞毒性、选择性和化疗增敏作用,同时通过荧光显微镜、流式细胞术和蛋白质印迹考察了亚细胞定位、ROS 生成和细胞死亡机制。通过 NIR 成像和 22Rv1 异种移植模型确定了肿瘤靶向和治疗疗效。
结果:在这些偶联物中,HMCD-ART1 显示出最强的效力和选择性,能有效杀伤对紫杉烷类、阿比特龙(abiraterone)和恩扎鲁胺(enzalutamide)耐药的前列腺癌细胞,同时不损伤正常前列腺上皮细胞。HMCD-ART1 增强了对化疗药物的敏感性,抑制了集落形成和迁移,并展现出长达八周的肿瘤滞留。机制分析揭示了线粒体和溶酶体共定位、ROS 诱导、细胞色素 c 释放、DNA 损伤(pATM、γH2AX)以及线粒体分裂依赖性坏死性死亡。在体内,HMCD-ART1 显著抑制了异种移植肿瘤生长,且未观察到全身毒性。
结论:HMCD-ART1 将 NIR 引导的肿瘤靶向与线粒体特异性细胞毒性相结合,提供了一个克服 mCRPC 耐药的新型治疗平台。该策略代表了一种结合靶向递送、亚细胞选择性和增强安全性的下一代精准化疗方法。
查看英文原文 English abstract
Background: Resistance to androgen-deprivation therapy leads to metastatic castration-resistant prostate cancer (mCRPC), a lethal stage with limited treatment options. Artemisinin (ART) derivatives exert anti-tumor activity through reactive oxygen species (ROS) generation and mitochondrial disruption but lack tumor selectivity. Our laboratory identified a near-infrared (NIR) heptamethine carbocyanine dye (HMCD) that preferentially accumulates in tumors via organic anion-transporting polypeptides (OATPs) upregulated under hypoxia, enabling tumor-specific delivery.
Methods: To achieve subcellularly targeted chemotherapy, four HMCD-ART conjugates with distinct linkers were synthesized: HMCD-ART1 (artesunate linker), HMCD-ART2 (carbamate linker), HMCD-ART3 (ether linker), and HMCD-ART4 (ester linker). These were evaluated in androgen-independent prostate cancer models (22Rv1, PC-3, C4-2B, and resistant derivatives). Cytotoxicity, selectivity, and chemosensitization were analyzed in vitro, while subcellular localization, ROS generation, and cell-death mechanisms were examined by fluorescence microscopy, flow cytometry, and Western blotting. Tumor targeting and therapeutic efficacy were determined by NIR imaging and 22Rv1 xenograft models.
Results: Among the conjugates, HMCD-ART1 showed the greatest potency and selectivity, effectively killing taxane-, abiraterone-, and enzalutamide-resistant prostate cancer cells while sparing normal prostate epithelial cells. HMCD-ART1 enhanced sensitivity to chemotherapeutics, inhibited colony formation and migration, and demonstrated prolonged tumor retention for up to eight weeks. Mechanistic analyses revealed mitochondrial and lysosomal co-localization, ROS induction, cytochrome c release, DNA damage (pATM, gammaH2AX), and mitochondrial-fission-dependent necrotic death. In vivo, HMCD-ART1 significantly suppressed xenograft tumor growth without observable systemic toxicity.
Conclusion: HMCD-ART1 integrates NIR-guided tumor targeting with mitochondrial-specific cytotoxicity, providing a novel therapeutic platform to overcome drug resistance in mCRPC. This strategy represents a next-generation precision chemotherapeutic approach combining targeted delivery, subcellular selectivity, and enhanced safety.
利益披露 Disclosure
Y. Ou, None..
A. Lim, None..
M. Edderkaoui, None..
R. Wang, None..
Q. Wang, None..
S. J. Pandol, None..
Y. Zhang, None.