Li Chunmou, Wen Yuchen, Wang Jiasheng, Li Lindi, He Yue, Cheng Yucai, Chen Junru, Huang Junbin, Ouyang Cheng, Liu Yong, Zhou Ruizhi, Chen Haisheng, Li Fei, Guo Qiqi, Chen Yun, Chen Chun, Zhang Qing
Pediatric Hematology Laboratory, Division of Hematology/Oncology, Department of Pediatrics, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, Guangdong, 518107, People's Republic of China.
State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, Guangdong, 510275, People's Republic of China.
Int J Nanomedicine. 2025 Apr 24;20:5267-5289. doi: 10.2147/IJN.S511713. eCollection 2025.
Acute myeloid leukemia (AML) is a highly heterogeneous disease with poor therapeutic outcomes and overall prognosis, particularly in c-Mpl+ AML. , a proto-oncogene, is expressed at significantly higher levels in AML compared to normal human tissue cells. This study aimed to develop a type of targeted exosomes (Exos) capable of delivering anticancer drugs directly to c-Mpl+ AML cells.
Human umbilical cord mesenchymal stem cells (hUCMSCs) were isolated as the source of Exos. Fusion CD63 proteins with varying numbers of thrombopoietin (TPO)-mimic peptides, designed to target c-Mpl, were bioengineered to be expressed on the membranes of hUCMSCs and their derived Exos. The targeting capability of the fusion proteins was assessed using the DUAL membrane system, fluorescence resonance energy transfer efficiency, and endocytosis assays. After encapsulating the anticancer drug daunorubicin (DNR), these targeted Exos were evaluated for their ability to eliminate c-Mpl+ AML cells. Safety and efficacy were further tested in a mouse AML model.
Our findings showed that the engineered hUCMSCs-derived Exos demonstrated excellent targeting ability to c-Mpl and a strong propensity for endocytic uptake by c-Mpl+ AML cells. Among the engineered Exos, those with the fusion protein containing three TPO-mimic peptides (CD63-mTPO), named as mExos, exhibited the highest binding affinity for c-Mpl. When loaded with DNR, these engineered Exos (mExos@DNR) effectively eliminated c-Mpl+ AML cells in both in vitro and in vivo experiments. Furthermore, safety assessments revealed that therapy-related toxicities were within acceptable limits and associated with manageable side effects.
In summary, our results suggest engineered Exos as a highly effective targeted drug delivery vehicle for eliminating c-Mpl+ AML cells while maintaining a favorable safety profile. These findings also provide valuable insights for developing therapeutic strategies for AML and other tumors characterized by specific membrane protein expression.
急性髓系白血病（AML）是一种高度异质性疾病，治疗效果和总体预后较差，尤其是在c-Mpl+ AML中。原癌基因 在AML中的表达水平明显高于正常人体组织细胞。本研究旨在开发一种能够将抗癌药物直接递送至c-Mpl+ AML细胞的靶向外泌体（Exos）。
分离人脐带间充质干细胞（hUCMSCs）作为Exos的来源。将具有不同数量血小板生成素（TPO）模拟肽的融合CD63蛋白进行生物工程改造，使其在hUCMSCs及其衍生的Exos膜上表达，这些模拟肽旨在靶向c-Mpl。使用双膜系统、荧光共振能量转移效率和内吞作用测定法评估融合蛋白的靶向能力。在包裹抗癌药物柔红霉素（DNR）后，评估这些靶向Exos清除c-Mpl+ AML细胞的能力。在小鼠AML模型中进一步测试安全性和有效性。
我们的研究结果表明，工程化的hUCMSCs衍生的Exos对c-Mpl具有优异的靶向能力，并且c-Mpl+ AML细胞具有强烈的内吞摄取倾向。在工程化的Exos中，含有三个TPO模拟肽（CD63-mTPO）的融合蛋白的Exos，命名为mExos，对c-Mpl表现出最高的结合亲和力。当负载DNR时，这些工程化的Exos（mExos@DNR）在体外和体内实验中均有效清除c-Mpl+ AML细胞。此外，安全性评估显示，治疗相关毒性在可接受范围内，且副作用可控。
总之，我们的结果表明工程化的Exos是一种高效的靶向药物递送载体，可在保持良好安全性的同时清除c-Mpl+ AML细胞。这些发现也为开发AML和其他以特定膜蛋白表达为特征的肿瘤的治疗策略提供了有价值的见解。
