The Seventh Affiliated Hospital, Sun Yat-Sen University, Shenzhen, Guangdong, P.R. China.
Shenzhen Key Laboratory of Chinese Medicine Active Substance Screening and Translational Research, Shenzhen, Guangdong, P.R. China.
Br J Haematol. 2024 Oct;205(4):1532-1545. doi: 10.1111/bjh.19722. Epub 2024 Aug 27.
Bone marrow endothelial progenitor cells (BM EPCs) are crucial in supporting haematopoietic regeneration, while the BM EPCs of haematological patients with chemotherapy-induced thrombocytopenia (CIT) are unavoidably damaged. Therefore, the present study aimed to examine the effect of thrombopoietin (TPO) on the recovery of BM EPCs of CIT patients and to identify the underlying mechanisms. The cell functions were determined by 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil)-acetylated low-density lipoprotein (Dil-Ac-LDL) uptake and fluorescein isothiocyanate (FITC)-labeled Ulex europaeus agglutinin-I (FITC-UEA-I) binding assay, as well as proliferation, migration and tube formation experiments. Endothelial cells were transfected with METTL16 lentivirus, followed by methylated RNA immunoprecipitation sequencing. Zebrafish with vascular defect was used as the in vivo model. TPO significantly improved the quantity and functions of BM EPCs from CIT patients in vitro and restored the subintestinal vein area of zebrafish with vascular defect in vivo. Mechanically, TPO enhanced the BM EPC functions through Akt signal mediated by METTL16, which was downregulated in BM EPCs of CIT patients and involved in the regulation of endothelial functions. The present study demonstrates that TPO improves the recovery of BM EPCs from CIT patients with haematological malignancies via METTL16/Akt signalling, which provides new insights into the role of TPO in treating CIT in addition to direct megakaryopoiesis.
骨髓内皮祖细胞(BM EPCs)在支持造血再生方面至关重要,而接受化疗诱导的血小板减少症(CIT)的血液病患者的 BM EPCs 不可避免地受损。因此,本研究旨在研究促血小板生成素(TPO)对 CIT 患者 BM EPC 恢复的影响,并确定其潜在机制。通过 1,1'-二辛基-3,3,3',3'-四甲基吲哚羰花青高氯酸盐(Dil)-乙酰化低密度脂蛋白(Dil-Ac-LDL)摄取和荧光素异硫氰酸酯(FITC)标记的欧洲菘蓝凝集素-I(FITC-UEA-I)结合测定、增殖、迁移和管形成实验来确定细胞功能。用 METTL16 慢病毒转染内皮细胞,然后进行甲基化 RNA 免疫沉淀测序。将血管缺陷的斑马鱼用作体内模型。TPO 显著改善了 CIT 患者体外 BM EPC 的数量和功能,并恢复了血管缺陷斑马鱼的亚肠静脉面积。在机制上,TPO 通过 METTL16 介导的 Akt 信号增强了 BM EPC 的功能,而 METTL16 在 CIT 患者的 BM EPC 中下调,并参与了内皮功能的调节。本研究表明,TPO 通过 METTL16/Akt 信号增强了血液恶性肿瘤 CIT 患者 BM EPC 的恢复,除了直接巨核细胞生成外,这为 TPO 在治疗 CIT 中的作用提供了新的见解。