Single‐cell RNA sequencing in facioscapulohumeral muscular dystrophy disease etiology and development
Single‐cell RNA sequencing in facioscapulohumeral muscular dystrophy disease etiology and development
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DOI：
10.1093/hmg/ddy400
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发表时间：
2018-11
影响因子：
3.5
通讯作者：
Anita van den Heuvel;A. Mahfouz;S. Kloet;J. Balog;B. van Engelen;R. Tawil;S. Tapscott;S. M. van der Maarel
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文献类型：
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作者：
Anita van den Heuvel;A. Mahfouz;S. Kloet;J. Balog;B. van Engelen;R. Tawil;S. Tapscott;S. M. van der Maarel
摘要面肩肱型肌营养不良症（FSHD）的特征是骨骼肌中转录因子DUX4的散发性去抑制。DUX4激活一系列肌肉破坏事件，最终导致肌肉萎缩和细胞凋亡。然而，零星DUX4表达如何导致全身性肌肉萎缩仍不清楚。转录组分析已经系统地受到大多数细胞核是DUX4阴性的挑战，削弱了DUX4转录组特征。此外，DUX4已被证明以高度动态的突发样方式表达，可能导致在DUX4表达本身消失很久之后检测到下游级联事件。因此，鉴定单个细胞中的FSHD转录组并解开导致FSHD发展的级联事件可能为疾病过程提供重要的见解。我们采用单细胞RNA测序，结合伪时间轨迹建模，研究FSHD疾病的病因和人类原代肌细胞的细胞进展。我们在所有测试的患者来源的培养物中鉴定了一个小的FSHD特异性细胞群，并检测到与DUX4去抑制相关的新基因。我们还生成了一个FSHD细胞进展模型，反映了早期爆发样DUX4表达以及各种FSHD相关途径的下游激活，这使我们能够将DUX4表达特征动态与调控复合物的动态相关联，从而促进DUX4沉默的表观遗传靶点的优先化。因此，单细胞转录组学结合伪时间建模可以提供有关FSHD疾病病因和进展的有价值的信息，这些信息可以潜在地指导治疗的生物标志物和靶标选择。
Abstract Facioscapulohumeral muscular dystrophy (FSHD) is characterized by sporadic de‐repression of the transcription factor DUX4 in skeletal muscle. DUX4 activates a cascade of muscle disrupting events, eventually leading to muscle atrophy and apoptosis. Yet, how sporadic DUX4 expression leads to the generalized muscle wasting remains unclear. Transcriptome analyses have systematically been challenged by the majority of nuclei being DUX4neg, weakening the DUX4 transcriptome signature. Moreover, DUX4 has been shown to be expressed in a highly dynamic burst‐like manner, likely resulting in the detection of the downstream cascade of events long after DUX4 expression itself has faded. Identifying the FSHD transcriptome in individual cells and unraveling the cascade of events leading to FSHD development may therefore provide important insights in the disease process. We employed single‐cell RNA sequencing, combined with pseudotime trajectory modeling, to study FSHD disease etiology and cellular progression in human primary myocytes. We identified a small FSHD‐specific cell population in all tested patient‐derived cultures and detected new genes associated with DUX4 de‐repression. We furthermore generated an FSHD cellular progression model, reflecting both the early burst‐like DUX4 expression as well as the downstream activation of various FSHD‐associated pathways, which allowed us to correlate DUX4 expression signature dynamics with that of regulatory complexes, thereby facilitating the prioritization of epigenetic targets for DUX4 silencing. Single‐cell transcriptomics combined with pseudotime modeling thus holds valuable information on FSHD disease etiology and progression that can potentially guide biomarker and target selection for therapy.