Department of Dermatology, The Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong, China.
Department of Clinical Laboratory, The Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong, China.
Skin Res Technol. 2023 Jan;29(1):e13259. doi: 10.1111/srt.13259.
Targeting CD20+ melanoma cancer stem cells (CSCs) subset is essential for treating melanoma. Anti-CD20 aptamer-modified exosomes (ACEXO) loaded with Adriamycin could be a therapeutic strategy for targeting CSCs.
Exosomes loaded with Adriamycin were modified with anti-CD20 aptamer and characterized by size and molecular markers using transmission electron microscope and dynamic light scattering. The uptake of ACEXO into CD20+ cells was checked, and its cytotoxicities in CD20+ melanoma cells, HEK 293T, and 3T3 cells were evaluated. At the same time, the in vivo distribution of ACEXO in the tumor-bearing mice model was determined.
The particle size of the exosome is about 80-100 nm. Western blot analysis showed that they expressed the characteristic exosome markers: CD9 and CD63. Quantitative analysis of the mean fluorescence intensity after 4 h incubation showed that ACEXO significantly improved Adriamycin uptake. Notably, the ACEXO killed only CD20+ melanoma cells. In addition, they exhibited good biocompatibility with both 293T and 3T3 cells at all doses. After intravenous injection, exosome distribution data showed that ACEXO's accumulation in the tumor is higher than anti-CD20-modified exosomes (AEXO)'s at all time points, and the accumulation increased as time prolonged. Addition of ACEXO reduces the number of tumorspheres in A375 or WM266-4 cells compared to untreated controls or AEXO-treated group. More important, while treating melanoma tumor-bearing mice, ACEXO-treated group showed the lowest tumor weight without body weight loss.
ACEXO loaded with Adriamycin could suppress tumor cell growth in vitro and in vivo, probably by targeting CD20+ melanoma CSCs.
靶向 CD20+黑色素瘤癌干细胞（CSC）亚群对于治疗黑色素瘤至关重要。载阿霉素的抗 CD20 适体修饰外泌体（ACEXO）可能是一种针对 CSC 的治疗策略。
用抗 CD20 适体修饰载阿霉素的外泌体，并通过透射电子显微镜和动态光散射检查其大小和分子标志物。检查 ACEXO 进入 CD20+细胞的摄取情况，并评估其对 CD20+黑色素瘤细胞、HEK 293T 和 3T3 细胞的细胞毒性。同时，确定 ACEXO 在荷瘤小鼠模型中的体内分布。
外泌体的粒径约为 80-100nm。Western blot 分析表明，它们表达了特征性的外泌体标记物：CD9 和 CD63。孵育 4 小时后定量分析平均荧光强度表明，ACEXO 显著提高了阿霉素的摄取。值得注意的是，ACEXO 仅杀死 CD20+黑色素瘤细胞。此外，它们在所有剂量下与 293T 和 3T3 细胞均表现出良好的生物相容性。静脉注射后，外泌体分布数据显示，ACEXO 在所有时间点在肿瘤中的积累均高于抗 CD20 修饰外泌体（AEXO），并且随着时间的延长积累增加。与未处理对照或 AEXO 处理组相比，添加 ACEXO 可减少 A375 或 WM266-4 细胞中的肿瘤球数量。更重要的是，在治疗黑色素瘤荷瘤小鼠时，ACEXO 处理组的肿瘤重量最低，而体重没有减轻。
载阿霉素的 ACEXO 可在体外和体内抑制肿瘤细胞生长，可能通过靶向 CD20+黑色素瘤 CSC。
