Fu Huihui, Xie Xiaoqing, Zhai Liuyue, Liu Yi, Tang Yifeng, He Sanxiu, Li Jun, Xiao Qing, Xu Guofa, Yang Zailin, Zhang Xiaomei, Liu Yao
Department of Hematology Oncology, Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, China.
Department of Hematology, Chongqing, Central Laboratory, Chongqing University Fuling Hospital, Chongqing University Fuling Hospital, Chongqing, China.
Stem Cell Res Ther. 2024 Dec 2;15(1):460. doi: 10.1186/s13287-024-04079-3.
Acute myeloid leukemia (AML) is characterized by abundant immature myeloid cells, relapse and refractory due to leukemia stem cells (LSCs). Bone marrow mesenchymal stem/ stromal cells (BMSCs) supported LSCs survival, meanwhile, chemotherapy improved connexin43 (CX43) expression. CX43, as the most intercellular gap junction, facilitated transmit mitochondria from BMSCs into AML. We hypothesized that increased mitochondria transferred from BMSCs supported metabolic remodeling in LSCs to sustain their stemness.
Primary BMSCs from AML patients were isolated. CX43-BMSCs, overexpressing CX43, were cocultured with KG-1a cells. Fluorescence and confocal microscopy observed mitochondrial transfer. Flow cytometry, EdU assay, and clonogenicity evaluated cell cycle, proliferation, and clonogenic potential. Xenograft mouse models were used to evaluate the tumorigenicity of KG-1a in vivo. Seahorse, RNA-seq, and LC-MS assessed mitochondrial function, transcriptomes, and metabolites post-coculture.
CX43-BMSCs promoted unidirectional mitochondrial transfer, enhancing KG-1a adhesion and proliferation to maintain LSCs stemness in vitro and vivo. RNA-seq revealed coculture with CX43-BMSCs upregulated genes related to adhesion, proliferation, and migration in KG-1a cells. Elevated CX43 expression strengthened BMSCs-KG-1a interaction, facilitating mitochondrial transfer and nucleoside metabolism, fueling KG-1a cells. This enhanced mitochondrial energy metabolism, promoting metabolic reprogramming and clonogenicity.
CX43-mediated mitochondrial transfer from BMSCs to KG-1a enhances LSCs adhesion, proliferation, clonogenicity, and metabolic reprogramming. CX43 emerges as a potential therapeutic target for AML by sustaining LSCs stemness through metabolic remodeling.
急性髓系白血病（AML）的特征是存在大量未成熟髓系细胞，由于白血病干细胞（LSCs）导致复发和难治。骨髓间充质干细胞（BMSCs）支持LSCs存活，同时，化疗可提高连接蛋白43（CX43）的表达。CX43作为最主要的细胞间缝隙连接，促进线粒体从BMSCs转移至AML细胞。我们推测，BMSCs转移来的线粒体增加支持了LSCs的代谢重塑以维持其干性。
分离AML患者的原代BMSCs。将过表达CX43的CX43-BMSCs与KG-1a细胞共培养。通过荧光和共聚焦显微镜观察线粒体转移。采用流式细胞术、EdU检测和克隆形成试验评估细胞周期、增殖和克隆形成潜力。利用异种移植小鼠模型评估KG-1a在体内的致瘤性。采用海马分析仪、RNA测序和液相色谱-质谱联用技术评估共培养后的线粒体功能、转录组和代谢产物。
CX43-BMSCs促进单向线粒体转移，增强KG-1a的黏附与增殖，从而在体外和体内维持LSCs的干性。RNA测序显示，与CX43-BMSCs共培养上调了KG-1a细胞中与黏附、增殖和迁移相关的基因。CX43表达升高增强了BMSCs与KG-1a的相互作用，促进线粒体转移和核苷代谢，为KG-1a细胞提供能量。这增强了线粒体能量代谢，促进代谢重编程和克隆形成。
CX43介导的线粒体从BMSCs转移至KG-1a增强了LSCs的黏附、增殖、克隆形成能力和代谢重编程。CX43通过代谢重塑维持LSCs干性，成为AML的一个潜在治疗靶点。
