中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2024, Vol. 19 ›› Issue (1): 205-211.doi: 10.4103/1673-5374.375347

• 原著:退行性病与再生 • 上一篇    下一篇

MORC2 p.S87L突变抑制相关信号通路导致脊髓肌萎缩样表型诱导多能干细胞增殖障碍

  

  • 出版日期:2024-01-15 发布日期:2023-08-02
  • 基金资助:
    国家自然科学基金项目（82171172和81771366），中南大学中央高校基本科研业务费（2021zzts1095和2022zzts0832）

The MORC2 p.S87L mutation reduces proliferation of pluripotent stem cells derived from a patient with the spinal muscular atrophy-like phenotype by inhibiting proliferation-related signaling pathways

Sen Zeng1, #, Honglan Yang1, #, Binghao Wang2, Yongzhi Xie3, Ke Xu1, Lei Liu1, 4, Wanqian Cao1, Xionghao Liu5, Beisha Tang6, Mujun Liu7, *, Ruxu Zhang1, *#br#   

  1. 1Department of Neurology, The Third Xiangya Hospital, Central South University, Changsha, Hunan Province, China; 2Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China; 3Department of Radiology, The Third Xiangya Hospital, Central South University, Changsha, Hunan Province, China; 4Health Management Center, The Third Xiangya Hospital, Central South University, Changsha, Hunan Province, China; 5Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha, Hunan Province, China; 6National Clinical Research Center for Geriatric Disorders, Central South University, Changsha, Hunan Province, China; 7Department of Cell Biology, School of Life Sciences, Central South University, Changsha, Hunan Province, China
  • Online:2024-01-15 Published:2023-08-02
  • Contact: Ruxu Zhang, MD, PhD, zhangruxu@vip.163.com; Mujun Liu, PhD, liumujun03@csu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, Nos. 82171172 (to RZ) and 81771366 (to RZ) and Fundamental Research Funds for the Central Universities of Central South University, Nos. 2021zzts1095 (to SZ) and 2022zzts0832 (to HY).

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摘要：

染色质重塑蛋白MORC家族CW型锌指结构蛋白2 (MORC2)是腓骨肌萎缩2Z型的致病基因，其热点突变p.S87L与更严重的脊髓性肌萎缩症样临床表型有关。为了解MORC2 p.S87L突变导致严重临床表型的潜在机制，实验收集了脊髓性肌萎缩样患者（MORC2 p.S87L）、腓骨肌萎缩2Z型患者（MOLC2 p.Q400R）以及健康对照的尿液上皮细胞，诱导产生多能干细胞，再分化为运动神经元前体。经RNA测序和KEGG富集分析发现，PI3K/Akt和MAPK/ERK信号通路在脊髓性肌萎缩样患者来源细胞中富集，且在多能干细胞阶段显著下调。与腓骨肌萎缩2Z型和健康对照来源细胞相比，脊髓性肌萎缩样患者来源细胞在多能干细胞和运动神经元前体阶段的增殖明显减少，且在多能干细胞阶段存在G0/G1期阻滞。在多能干细胞阶段中以特异性反义寡核苷酸沉默MORC2 p.S87L，可见的改善细胞增殖和PI3K/Akt和MAPK/ERK通路活化明显恢复，但在运动神经元前体阶段干预则对细胞增殖无显著效果。上述发现表明，多能干细胞阶段细胞增殖减少和G0/G1期阻滞可能是通过下调PI3K/Akt和MAPK/ERK信号通路导致了严重的脊髓性肌萎缩症样表型，且早期靶向MORC2 p.S87L突变的反义寡核苷酸基因沉默治疗能够改善诱导性多能干细胞阶段的增殖障碍。

https://orcid.org/0000-0003-3823-8148 (Ruxu Zhang); https://orcid.org/0000-0002-1558-1448 (Mujun Liu)

关键词: 脊髓肌萎缩样, 腓骨肌萎缩2Z型, 诱导多能干细胞, 细胞增殖, 细胞周期阻滞, PI3K/Akt, MAPK/ERK, 反义寡核苷酸

Abstract:

Mutations in the microrchidia CW-type zinc finger protein 2 (MORC2) gene are the causative agent of Charcot-Marie-Tooth disease type 2Z (CMT2Z), and the hotspot mutation p.S87L is associated with a more severe spinal muscular atrophy-like clinical phenotype. The aims of this study were to determine the mechanism of the severe phenotype caused by the MORC2 p.S87L mutation and to explore potential treatment strategies. Epithelial cells were isolated from urine samples from a spinal muscular atrophy (SMA)-like patient (MORC2 p.S87L), a CMT2Z patient (MORC2 p.Q400R), and a healthy control and induced to generate pluripotent stem cells, which were then differentiated into motor neuron precursor cells. Next-generation RNA sequencing followed by KEGG pathway enrichment analysis revealed that differentially expressed genes involved in the PI3K/Akt and MAPK/ERK signaling pathways were enriched in the p.S87L SMA-like patient group and were significantly downregulated in induced pluripotent stem cells. Reduced proliferation was observed in the induced pluripotent stem cells and motor neuron precursor cells derived from the p.S87L SMA-like patient group compared with the CMT2Z patient group and the healthy control. G0/G1 phase cell cycle arrest was observed in induced pluripotent stem cells derived from the p.S87L SMA-like patient. MORC2 p.S87L-specific antisense oligonucleotides (p.S87L-ASO-targeting) showed significant efficacy in improving cell proliferation and activating the PI3K/Akt and MAPK/ERK pathways in induced pluripotent stem cells. However, p.S87L-ASO-targeting did not rescue proliferation of motor neuron precursor cells. These findings suggest that downregulation of the PI3K/Akt and MAPK/ERK signaling pathways leading to reduced cell proliferation and G0/G1 phase cell cycle arrest in induced pluripotent stem cells might be the underlying mechanism of the severe p.S87L SMA-like phenotype. p.S87L-ASO-targeting treatment can alleviate disordered cell proliferation in the early stage of pluripotent stem cell induction. 

Key words: antisense oligonucleotides, cell cycle arrest, Charcot-Marie-Tooth disease 2Z, induced pluripotent stem cells, MAPK/ERK, PI3K/Akt, proliferation, spinal muscular atrophy-like