Institute of Biopathology and Regenerative Medicine (IBIMER), Center of Biomedical Research (CIBM), University of Granada, Granada, 18100, Spain.
Department of Anatomy and Embryology, Faculty of Medicine, University of Granada, Granada, 18071, Spain.
Int J Nanomedicine. 2024 Feb 23;19:1843-1865. doi: 10.2147/IJN.S440881. eCollection 2024.
The lack of specificity of conventional chemotherapy is one of the main difficulties to be solved in cancer therapy. Biomimetic magnetoliposomes are successful chemotherapy controlled-release systems, hyperthermia, and active targeting agents by functionalization of their surface with monoclonal antibodies. The membrane receptor Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) stands out as colorectal cancer (CRC) biomarker and appears to be related to treatment resistance and the development of metastasis. The aim of this study was to assess the effectiveness and safety of LGR5-targeted biomimetic magnetoliposomes loaded with oxaliplatin (OXA) or 5-fluorouracil (5-FU) in the selective treatment of CRC and their possible application in hyperthermia.
Synthesis, characterization and determination of heating capacity of magnetoliposomes transporting OXA or 5-FU (with and without LGR5 functionalization) were conducted. In vitro antitumoral activity was assayed in multiple colorectal cell lines at different times of exposition. In addition to this, cell internalization was studied by Prussian Blue staining, flow cytometry and fluorescence microscopy. In vivo acute toxicity of magnetoliposomes was performed to evaluate iron-related toxicity.
OXA and 5-FU loaded magnetoliposomes functionalized with LGR5 antibody showed higher cellular uptake than non-targeted nanoformulation with a reduction of the percentage of proliferation in colon cancer cell lines up to 3.2-fold of the IC value compared to that of free drug. The differences between non-targeted and targeted nanoformulations were more evident after short exposure times (4 and 8 hours). Interestingly, assays in the MC38 transduced cells with reduced LGR5 expression (MC38-L(-)), showed lower cell internalization of LGR5-targeted magnetoliposomes compared to non-transduced MC38 cell line. In addition, magnetoliposomes showed an in vitro favorable heating response under magnetic excitation and great iron-related biocompatibility data in vivo.
Drug-loaded magnetoliposomes functionalized with anti-LGR5 antibodies could be a promising CRC treatment strategy for LGR5+ targeted chemotherapy, magnetic hyperthermia, and both in combination.
常规化疗缺乏特异性，是癌症治疗中需要解决的主要难题之一。仿生磁脂质体是通过表面功能化单克隆抗体实现化疗药物控释、热疗和主动靶向的成功载体。富含亮氨酸重复序列的 G 蛋白偶联受体 5（LGR5）作为结直肠癌（CRC）的生物标志物脱颖而出，并且似乎与治疗耐药和转移的发展有关。本研究旨在评估载奥沙利铂（OXA）或 5-氟尿嘧啶（5-FU）的 LGR5 靶向仿生磁脂质体在 CRC 选择性治疗中的有效性和安全性，以及它们在热疗中的可能应用。
合成、表征和测定载 OXA 或 5-FU 的磁脂质体（带和不带 LGR5 功能化）的加热能力。在不同暴露时间的多种结直肠癌细胞系中检测体外抗肿瘤活性。此外，通过普鲁士蓝染色、流式细胞术和荧光显微镜研究细胞内化。进行磁脂质体的体内急性毒性实验以评估铁相关毒性。
载 LGR5 抗体的 OXA 和 5-FU 负载磁脂质体的细胞摄取率高于非靶向纳米制剂，与游离药物相比，在结肠癌细胞系中增殖的百分比降低了 3.2 倍。与非靶向纳米制剂相比，短暴露时间（4 和 8 小时）下，非靶向和靶向纳米制剂之间的差异更为明显。有趣的是，在 LGR5 表达降低的 MC38 转导细胞（MC38-L（-））中进行的测定显示，与非转导的 MC38 细胞系相比，LGR5 靶向磁脂质体的细胞内化率较低。此外，磁脂质体在磁场激发下表现出良好的体外加热响应，并且在体内具有很好的铁相关生物相容性数据。
载药磁脂质体经抗 LGR5 抗体功能化后，可能成为 LGR5+靶向化疗、磁热疗及两者联合治疗 CRC 的一种有前途的治疗策略。
