鉴定 Duchenne 肌营养不良症的枢纽基因和治疗性 siRNA，开发新的辅助治疗方法。

Identification of hub genes and therapeutic siRNAs to develop novel adjunctive therapy for Duchenne muscular dystrophy.

机构信息

Department of Aerospace Medical Training, School of Aerospace Medicine, Air Force Medical University, Xi'an, China.

School of Sports Science, Beijing Sport University, Beijing, China.

出版信息

BMC Musculoskelet Disord. 2024 May 18;25(1):386. doi: 10.1186/s12891-024-07206-6.

DOI:10.1186/s12891-024-07206-6

PMID:38762732

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11102231/

Abstract

OBJECTIVE

Duchenne muscular dystrophy (DMD) is a devastating X-linked neuromuscular disorder caused by various defects in the dystrophin gene and still no universal therapy. This study aims to identify the hub genes unrelated to excessive immune response but responsible for DMD progression and explore therapeutic siRNAs, thereby providing a novel treatment.

METHODS

Top ten hub genes for DMD were identified from GSE38417 dataset by using GEO2R and PPI networks based on Cytoscape analysis. The hub genes unrelated to excessive immune response were identified by GeneCards, and their expression was further verified in mdx and C57 mice at 2 and 4 months (M) by (RT-q) PCR and western blotting. Therapeutic siRNAs were deemed as those that could normalize the expression of the validated hub genes in transfected C2C12 cells.

RESULTS

855 up-regulated and 324 down-regulated DEGs were screened from GSE38417 dataset. Five of the top 10 hub genes were considered as the candidate genes unrelated to excessive immune response, and three of these candidates were consistently and significantly up-regulated in mdx mice at 2 M and 4 M when compared with age-matched C57 mice, including Col1a2, Fbn1 and Fn1. Furthermore, the three validated up-regulated candidate genes can be significantly down-regulated by three rational designed siRNA (p < 0.0001), respectively.

CONCLUSION

COL1A2, FBN1 and FN1 may be novel biomarkers for DMD, and the siRNAs designed in our study were help to develop adjunctive therapy for Duchenne muscular dystrophy.

摘要

目的

杜氏肌营养不良症（DMD）是一种由肌营养不良蛋白基因突变引起的毁灭性 X 连锁神经肌肉疾病，目前尚无通用疗法。本研究旨在确定与过度免疫反应无关但与 DMD 进展有关的关键基因，并探索治疗性 siRNA，从而提供新的治疗方法。

方法

通过 GEO2R 和 Cytoscape 分析中的 PPI 网络，从 GSE38417 数据集确定 DMD 的前 10 个关键基因。通过 GeneCards 确定与过度免疫反应无关的关键基因，并通过（RT-q）PCR 和 Western blot 在 2 个月（M）和 4 个月（M）的 mdx 和 C57 小鼠中进一步验证其表达。将能够使转染的 C2C12 细胞中验证的关键基因表达正常化的治疗性 siRNA 视为有效 siRNA。

结果

从 GSE38417 数据集筛选出 855 个上调和 324 个下调的 DEGs。前 10 个关键基因中有 5 个被认为是与过度免疫反应无关的候选基因，其中 3 个候选基因在与年龄匹配的 C57 小鼠相比，在 2 M 和 4 M 时在 mdx 小鼠中持续且显著上调，包括 Col1a2、Fbn1 和 Fn1。此外，这三个验证的上调候选基因可以分别被三个合理设计的 siRNA 显著下调（p<0.0001）。