1例脊髓延髓肌萎缩症运动神经元分化及分子病理分析

doi:10.6040/j.issn.1671-7554.0.2024.0118

摘要/Abstract

摘要： 目的运用脊髓延髓肌萎缩症(spinal and bulbar muscular atrophy, SBMA)患者诱导多潜能干细胞(induced pluripotent stem cells, iPSCs)来源的运动神经元探究本病的分子病理表型。方法提取1例SBMA患者及1例健康对照者的皮肤成纤维细胞并重编程为iPSCs;将iPSCs诱导为运动神经前体细胞(motor neurons progenitor, MNP)并分化为成熟的运动神经元(mature motor neurons, mMN);免疫荧光染色研究细胞的增殖与凋亡,检测细胞核内包涵体的形成。结果患者,35岁,男性,主要表现为四肢近端无力、手抖以及性功能减退。骨骼肌活检显示神经源性损害病理改变。外周血毛细管电泳检测显示雄激素受体(androgen receptor, AR)基因第1外显子CAG重复次数为44次。iPSCs均表达干性标记物,细胞核型正常,并能随机分化为三胚层细胞类型。患者iPSCs及MNP未见细胞核内包涵体,但mMN中的核内包涵体较为典型。与健康对照相比,患者MNP增殖比例下降,凋亡比例增加。结论 SBMA患者iPSCs携带致病变异,其分化的mMN含有典型的细胞核内包涵体,精准地模拟本病的分子病理特征,为后续机制研究和药物靶标发现提供理想的模型。

关键词: 脊髓延髓肌萎缩症, 肯尼迪病, 雄激素受体基因, 诱导多潜能干细胞, 运动神经元

Abstract: Objective To explore the molecular pathologic phenotype of spinal and bulbar muscular atrophy(SBMA)using motor neurons derived from induced pluripotent stem cells(iPSCs)from SBMA patients. Methods Skin fibroblasts from one SBMA patient and one healthy control were extracted and reprogrammed into iPSCs; iPSCs were induced into motor neurons progenitor(MNP)and differentiated into mature motor neurons(mMN); immunofluorescence staining was used to study cell proliferation and apoptosis, and the formation of intranuclear inclusion bodies was detected. Results The patient was a 35-year-old male who presented with proximal weakness of the extremities, hand tremors, and hypogonadism. Skeletal muscle biopsy showed pathologic changes of neurogenic damage. Peripheral blood capillary electrophoresis showed 44 repeats of CAG in exon 1 of the AR gene. iPSCs all expressed stemness markers, had normal karyotypes, and were able to differentiate randomly into trichodermal cell types. No intranuclear inclusion bodies were seen in patient iPSCs and MNP, but intranuclear inclusion bodies were more typical in mMN. Compared with normal controls, the patients MNP had an increased proportion of apoptosis and a decreased proportion of proliferation. Conclusion The iPSCs of SBMA patients carried pathogenic mutations, and their differentiated mMNs contained typical intranuclear inclusion bodies, which accurately mimicked the molecular pathological features of the disease and provided an ideal model for the subsequent mechanism study and drug target discovery.

Key words: Spinal and Bulbar Muscular Atrophy, Kennedys Disease, Androgen Receptor Gene, Induced Pluripotent Stem Cells, Motor Neurons

中图分类号:

R746.4

刘文竹,刘付臣. 1例脊髓延髓肌萎缩症运动神经元分化及分子病理分析[J]. 山东大学学报 (医学版), 2024, 62(4): 92-100.

LIU Wenzhu, LIU Fuchen. Differentiation and molecular pathological analysis of motor neurons in a case of spinal muscular atrophy[J]. Journal of Shandong University (Health Sciences), 2024, 62(4): 92-100.

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