School of Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India.
School of Biotechnology, Jawaharlal Nehru University, New Delhi 110067, India.
Biochem Pharmacol. 2024 May;223:116199. doi: 10.1016/j.bcp.2024.116199. Epub 2024 Apr 9.
GNEM (GNE Myopathy) is a rare neuromuscular disease caused due to biallelic mutations in sialic acid biosynthetic GNE enzyme (UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine Kinase). Recently direct or indirect role of GNE in other cellular functions have been elucidated. Hyposialylation of IGF-1R leads to apoptosis due to mitochondrial dysfunction while hyposialylation of β1 integrin receptor leads to altered F-actin assembly, disrupted cytoskeletal organization and slow cell migration. Other cellular defects in presence of GNE mutation include altered ER redox state and chaperone expression such as HSP70 or PrdxIV. Currently, there is no cure to treat GNEM. Possible therapeutic trials focus on supplementation with sialic acid, ManNAc, sialyllactose and gene therapy that slows the disease progression. In the present study, we analyzed the effect of small molecules like BGP-15 (HSP70 modulator), IGF-1 (IGF-1R ligand) and CGA (cofilin activator) on cellular phenotypes of GNE heterozygous knock out L6 rat skeletal muscle cell line (SKM‑GNEHz). Treatment with BGP-15 improved GNE epimerase activity by 40 % and reduced ER stress by 45 % for SKM‑GNEHz. Treatment with IGF-1 improved epimerase activity by 37.5 %, F-actin assembly by 100 %, cell migration upto 36 % (36 h) and atrophy by 0.44-fold for SKM‑GNEHz. Treatment with CGA recovered epimerase activity by 49 %, F-actin assembly by 132 % and cell migration upto 41 % (24 h) in SKM‑GNEHz. Our study shows that treatment with these small effector molecules reduces the detrimental phenotype observed in SKM‑GNEHz, thereby, providing insights into potential therapeutic targets for GNEM.
GNE 肌病(GNEM)是一种罕见的神经肌肉疾病,由唾液酸生物合成 GNE 酶(UDP-N-乙酰氨基葡萄糖 2-差向异构酶/N-乙酰甘露糖胺激酶)的双等位基因突变引起。最近,GNE 在其他细胞功能中的直接或间接作用已经阐明。IGF-1R 的低唾液酸化导致线粒体功能障碍引起细胞凋亡,而β1 整合素受体的低唾液酸化导致 F-肌动蛋白组装改变、细胞骨架组织破坏和细胞迁移缓慢。在存在 GNE 突变的情况下,其他细胞缺陷包括 ER 氧化还原状态和伴侣蛋白表达的改变,如 HSP70 或 PrdxIV。目前,尚无治疗 GNEM 的方法。可能的治疗试验集中在补充唾液酸、ManNAc、唾液乳糖和基因治疗上,这些治疗方法可以减缓疾病进展。在本研究中,我们分析了小分子如 BGP-15(HSP70 调节剂)、IGF-1(IGF-1R 配体)和 CGA(丝切蛋白激活剂)对 GNE 杂合敲除 L6 大鼠骨骼肌细胞系(SKM-GNEHz)的细胞表型的影响。BGP-15 处理可使 SKM-GNEHz 的 GNE 差向异构酶活性提高 40%,内质网应激减少 45%。IGF-1 处理可使差向异构酶活性提高 37.5%,F-肌动蛋白组装提高 100%,细胞迁移提高 36%(36 小时),SKM-GNEHz 的萎缩减少 0.44 倍。CGA 处理可使 SKM-GNEHz 的差向异构酶活性恢复 49%,F-肌动蛋白组装提高 132%,细胞迁移提高 41%(24 小时)。我们的研究表明,这些小分子效应物的治疗可减少 SKM-GNEHz 中观察到的有害表型,从而为 GNEM 的潜在治疗靶点提供了见解。