Pharmacodynamic Studies of Fluorescent Diamond Carriers of Doxorubicin in Liver Cancer Cells and Colorectal Cancer Organoids.
Pharmacodynamic Studies of Fluorescent Diamond Carriers of Doxorubicin in Liver Cancer Cells and Colorectal Cancer Organoids.
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肝癌细胞和大肠癌器官中阿霉素的荧光钻石载体的药效学研究。
DOI：
10.2147/nsa.s321725
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Feuerstein GZ
中科院分区：
文献类型：
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作者：
Firestein R;Marcinkiewicz C;Nie L;Chua HK;Velazquez Quesada I;Torelli M;Sternberg M;Gligorijevic B;Shenderova O;Schirhagl R;Feuerstein GZ
我们最近报道了裸露荧光金刚石颗粒 FDP-NV-700/800nm (FDP-NV) 在大鼠静脉注射后优先沉积在肝脏中。 FDP-NV 在该物种中的药代动力学表明，通过肝脏的快速清除，其在循环中的停留时间较短。 FDP-NV 在肝脏中的滞留与任何病理学无关。这些观察结果表明，与 FDP-NV 相关的癌症治疗药物，例如阿霉素，可以潜在地用于抗癌治疗，同时避免外周器官的毒性。旨在对阿霉素包被的 FDP-NV-700/800nm (FDP-DOX) 作为转移性肝癌的前瞻性化疗药物进行概念验证 (POC) 和详细作用机制。 FDP-DOX 通过吸附化学产生。实验设计包括在体外肝癌细胞模型中与初始（裸）FDP-NV 进行比较的浓度和时间依赖性功效研究。通过流式细胞术和荧光显微镜证实了 HepG-2、Hep-3B 和 hCRC 类器官对 FDP-NV 和 FDP-DOX 的摄取。 FDP-DOX 药效作用包括代谢以及细胞死亡生物标志物膜联蛋白 V、TUNEL 和 LDH 渗漏。通过共聚焦显微镜和细胞组分的化学测定来评估从 FDP-DOX 解吸的 DOX。 FDP-DOX 的功效具有剂量和时间依赖性，并且在肝癌细胞系和人类 CRC 类器官中均有体现。 FDP-DOX 迅速内化到癌细胞/类器官中，导致癌症生长抑制和细胞凋亡。 FDP-DOX 破坏了细胞膜完整性，LDH 释放和抑制线粒体代谢途径（AlamarBlue 测定）证明了这一点。通过直接紫外-可见荧光测定和 DOX 荧光共聚焦显微镜证实游离 DOX 进入细胞核。在临床相关癌症模型中使用 FDP-DOX 观察到的快速吸收和深刻的癌症抑制作用，凸显了 FDP-DOX 在癌症化疗方面的前景。我们还得出结论，体外数据证明进一步投资体内 POC 研究是合理的。
We recently reported on preferential deposition of bare fluorescent diamond particles FDP-NV-700/800nm (FDP-NV) in the liver following intravenous administration to rats. The pharmacokinetics of FDP-NV in that species indicated short residency in the circulation by rapid clearance by the liver. Retention of FDP-NV in the liver was not associated with any pathology. These observations suggested that cancer therapeutics, such as doxorubicin, linked to FDP-NV, could potentially serve for anti-cancer treatment while sparing toxicities of peripheral organs. To generate proof-of-concept (POC) and detail mechanisms of action of doxorubicin-coated FDP-NV-700/800nm (FDP-DOX) as a prospective chemotherapeutic for metastatic liver cancer. FDP-DOX was generated by adsorption chemistry. Experimental design included concentration and time-dependent efficacy studies as compared with naïve (baren) FDP-NV in in vitro liver cancer cells models. Uptake of FDP-NV and FDP-DOX by HepG-2, Hep-3B and hCRC organoids were demonstrated by flow-cytometry and fluorescent microscopy. FDP-DOX pharmacodynamic effects included metabolic as well as cell death biomarkers Annexin V, TUNEL and LDH leakage. DOX desorpted from FDP-DOX was assessed by confocal microscopy and chemical assay of cells fractions. FDP-DOX efficacy was dose- and time-dependent and manifested in both liver cancer cell lines and human CRC organoids. FDP-DOX was rapidly internalized into cancer cells/organoids leading to cancer growth inhibition and apoptosis. FDP-DOX disrupted cell membrane integrity as evident by LDH release and suppressing mitochondrial metabolic pathways (AlamarBlue assay). Access of free DOX to the nuclei was confirmed by direct UV-Visible fluorescent assay and confocal microscopy of DOX fluorescence. The rapid uptake and profound cancer inhibition observed using FDP-DOX in clinically relevant cancer models, highlight FDP-DOX promise for cancer chemotherapeutics. We also conclude that the in vitro data justify further investment in in vivo POC studies.