Louwerse Max, Bila Kateryna O, van der Lienden Martijn J C, de Beaufort Arnout Jan M, Boot Rolf G, Artola Marta, van Eijk Marco, Aerts Johannes M F G
Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University, 2333 CC Leiden, The Netherlands.
Int J Mol Sci. 2025 Mar 18;26(6):2726. doi: 10.3390/ijms26062726.
Macrophages are specialised cells that degrade a range of substrates during their lifetime. In inherited lysosomal storage disorders, particularly the sphingolipidoses, macrophages transform into storage cells and contribute to pathology. An appropriate cultured macrophage model is desired for fundamental research and the assessment of considered therapeutic interventions. We compared commonly used macrophage cell lines, RAW264.7, J774A.1, and THP-1 cells, with human monocyte-derived macrophages (HMDMs) isolated from peripheral blood. Specific lysosomal glucosidases were analysed by enzymatic activity measurements and visualised with fluorescent activity-based probes. Special attention was given to lysosomal glucocerebrosidase (GBA1), the enzyme deficient in Gaucher disease in which lipid-laden macrophages are a hallmark. In macrophage cell lines and HMDMs, various (glyco)sphingolipids relevant to GBA1 activity were determined. Finally, the feasibility of inactivation of GBA1 with a cell-permeable suicide inhibitor was established, as well as the monitoring of uptake of therapeutic recombinant human GBA1. Major differences among various cell lines were noted in terms of morphology, lysosomal enzyme expression, and glycosphingolipid content. HMDMs appear to be the most suitable model for investigations into GBA1 and Gaucher disease. Moreover, they serve as a valuable model for mannose-receptor mediated uptake of therapeutic human GBA1, effectively mimicking enzyme replacement therapy for Gaucher disease.
巨噬细胞是一种特殊的细胞，在其生命周期内可降解多种底物。在遗传性溶酶体贮积症，尤其是鞘脂贮积症中，巨噬细胞会转变为贮积细胞并导致病理变化。基础研究和评估相关治疗干预措施都需要合适的巨噬细胞培养模型。我们将常用的巨噬细胞系RAW264.7、J774A.1和THP-1细胞与从外周血中分离的人单核细胞衍生巨噬细胞（HMDM）进行了比较。通过酶活性测量分析了特定的溶酶体葡萄糖苷酶，并用基于荧光活性的探针进行了可视化。特别关注了溶酶体葡萄糖脑苷脂酶（GBA1），该酶在戈谢病中缺乏，其中富含脂质的巨噬细胞是一个标志。在巨噬细胞系和HMDM中，测定了与GBA1活性相关的各种（糖）鞘脂。最后，确定了用细胞可渗透的自杀抑制剂使GBA1失活的可行性，以及监测治疗性重组人GBA1的摄取情况。在形态、溶酶体酶表达和糖鞘脂含量方面，各细胞系之间存在主要差异。HMDM似乎是研究GBA1和戈谢病最合适的模型。此外，它们还可作为甘露糖受体介导的治疗性人GBA1摄取的有价值模型，有效模拟戈谢病的酶替代疗法。
