271P 探索表观药物雷莫德林对小鼠成肌细胞分化的影响

IF 2.7 4区医学 Q2 CLINICAL NEUROLOGY

Neuromuscular Disorders Pub Date : 2024-10-01 DOI:10.1016/j.nmd.2024.07.089

V. Sian , J. Sarparanta , A. Hentschel , P. Jonson , A. Roos , S. Natraj Gayathri , B. Udd , M. Savarese , A. Nebbioso

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引用次数: 0

摘要

骨骼肌分化是一个严格控制和精心设计的过程，包括肌母细胞的排列和融合，以形成成熟的肌管。表观遗传学参与引导肌肉分化的兴趣日益浓厚，这引起了人们对表观调控因子作为这一过程中关键调控因子的关注。我们的体外研究旨在探讨雷莫德林在肌肉分化中的潜在作用。雷莫德林是 N-乙酰转移酶 10 的抑制剂，而 N-乙酰转移酶 10 是 RNA 乙酰化的关键角色。我们利用在超顺应性明胶水凝胶上培养的小鼠 C2C12 细胞的良好固化模型进行长期研究。新开发的水凝胶支架促进了肌管的排列和成熟，模拟了在体内观察到的骨骼肌生物学特性。我们在低血清条件下对 C2C12 细胞进行了长达 16 天的分化，并用表观药物 Remodelin 对其进行了处理。在分化的第7天，共聚焦图像显示，与对照组相比，雷莫德林导致肌管缺乏组织、形态不正常和成熟。即使在分化后期，经雷莫德林处理后也看不到明显的抽搐。雷莫德林下调了分化蛋白标志物的表达，但没有明显的表观调节作用。转录组学和蛋白质组学分析证实，雷莫德林有效地减缓了肌管的形成过程。RNAseq分析显示，表观药物下调了749个基因，这些基因主要编码涉及肌肉收缩、肌节组织、肌肉结构发育和钙离子结合的蛋白质。蛋白质组学分析进一步揭示，37 个明显下调的蛋白质与细胞外基质、细胞-基质粘附、肌肉细胞分化的正调控和钙离子结合有关。综上所述，这些结果表明雷莫德林对骨骼肌分化的调控网络具有广泛的影响。

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271P Exploring the effects of the epi-drug Remodelin on murine myoblasts differentiation

Skeletal muscle differentiation is a tightly controlled and elaborated process, that consists of alignment and fusion of myoblasts to form mature myotubes. A growing interest for epigenetics involvement in guiding muscle differentiation raised the attention on epi-modulators as pivotal regulators in this process. Our in vitro study aimed to investigate the potential effects of Remodelin in muscle differentiation. Remodelin is the inhibitor of N-acetyltransferase 10, the key-player of RNA acetylation. We used the well-consolidated model of murine C2C12 cells cultivated on ultra-compliant gelatin hydrogels for long-term studies. The newly developed hydrogel scaffold promotes myotube alignment and maturation, mimicking the skeletal muscle biology observed in vivo. We differentiated C2C12 cells in low-serum conditions up to 16 days and treated them with the epi-drug Remodelin. At day 7 of differentiation, confocal images revealed that Remodelin prompted a lack of organization, proper morphology, and maturation of myotubes compared to the control. Marked twitching was neither visible upon Remodelin treatment, even in the late stage of differentiation. Remodelin downregulated the expression of protein markers of differentiation, but without any significant epi-modulation. Both transcriptomics and proteomics analyses confirmed that Remodelin effectively slowed down the process of myotube formation. RNAseq analysis revealed that the epi-drug down-regulated 749 genes, predominantly encoding proteins involved in muscle contraction, sarcomere organization, muscle structure development, and calcium ion binding. Proteomics analysis further unveiled that 37 significantly down-regulated proteins were related to extracellular matrix, cell-matrix adhesion, positive regulation of muscle cell differentiation, and calcium ion binding. Taken together, these results suggest that Remodelin exerts a broad effect on the regulatory networks of skeletal muscle differentiation.

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来源期刊

Neuromuscular Disorders 医学-临床神经学

CiteScore

4.60

自引率

3.60%

发文量

543

审稿时长

53 days

期刊介绍： This international, multidisciplinary journal covers all aspects of neuromuscular disorders in childhood and adult life (including the muscular dystrophies, spinal muscular atrophies, hereditary neuropathies, congenital myopathies, myasthenias, myotonic syndromes, metabolic myopathies and inflammatory myopathies). The Editors welcome original articles from all areas of the field: • Clinical aspects, such as new clinical entities, case studies of interest, treatment, management and rehabilitation (including biomechanics, orthotic design and surgery). • Basic scientific studies of relevance to the clinical syndromes, including advances in the fields of molecular biology and genetics. • Studies of animal models relevant to the human diseases. The journal is aimed at a wide range of clinicians, pathologists, associated paramedical professionals and clinical and basic scientists with an interest in the study of neuromuscular disorders.