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Abstract：
Periostin, a multifunctional 90 kDa protein, plays a pivotal role in the pathogenesis of fibrosis across various tissues, including skeletal muscle. It operates within the transforming growth factor beta 1 (Tgf-β1) signalling pathway and is upregulated in fibrotic tissue. Alternative splicing of Periostin\'s C-terminal region leads to six protein-coding isoforms. This study aimed to elucidate the contribution of the isoforms containing the amino acids encoded by exon 17 (e17+ Periostin) to skeletal muscle fibrosis and investigate the therapeutic potential of manipulating exon 17 splicing. We identified distinct structural differences between e17+ Periostin isoforms, affecting their interaction with key fibrotic proteins, including Tgf-β1 and integrin alpha V. In vitro mouse fibroblast experimentation confirmed the TGF-β1-induced upregulation of e17+ Periostin mRNA, mitigated by an antisense approach that induces the skipping of exon 17 of the Postn gene. Subsequent in vivo studies in the D2.mdx mouse model of Duchenne muscular dystrophy (DMD) demonstrated that our antisense treatment effectively reduced e17+ Periostin mRNA expression, which coincided with reduced full-length Periostin protein expression and collagen accumulation. The grip strength of the treated mice was rescued to the wild-type level. These results suggest a pivotal role of e17+ Periostin isoforms in the fibrotic pathology of skeletal muscle and highlight the potential of targeted exon skipping strategies as a promising therapeutic approach for mitigating fibrosis-associated complications.
摘要：
骨膜素,一种多功能的90kDa蛋白质,在各种组织的纤维化发病机制中起着关键作用，包括骨骼肌.它在转化生长因子β1（Tgf-β1）信号传导途径中起作用，并在纤维化组织中上调。PeriostinC末端区域的可变剪接导致六种蛋白质编码亚型。这项研究旨在阐明包含外显子17（e17Periostin）编码的氨基酸的同工型对骨骼肌纤维化的贡献，并研究操纵外显子17剪接的治疗潜力。我们确定了e17+骨膜素同工型之间明显的结构差异，影响它们与关键纤维化蛋白的相互作用，包括Tgf-β1和整合素αV。体外小鼠成纤维细胞实验证实了TGF-β1诱导的e17骨膜素mRNA的上调，通过诱导Postn基因外显子17跳跃的反义方法减轻。随后的体内研究在D2。杜氏肌营养不良症（DMD）的mdx小鼠模型证明我们的反义治疗有效地降低了e17+骨膜素mRNA的表达，这与全长骨膜素蛋白表达和胶原蛋白积累减少相吻合。将处理的小鼠的握力恢复至野生型水平。这些结果表明e17+骨膜素同工型在骨骼肌纤维化病理中的关键作用，并突出了靶向外显子跳跃策略作为减轻纤维化相关并发症的有希望的治疗方法的潜力。
