酵母产生的重组人溶酶体β-己糖氨酸酶有效地挽救了Tay-Sachs病中GM2神经节苷脂的积累。

IF 3 3区 医学 Q2 HEALTH CARE SCIENCES & SERVICES
Orhan Kerim Inci, Andrés Felipe Leal, Nurselin Ates, Diego A Súarez, Angela Johana Espejo-Mojica, Carlos Javier Alméciga-Diaz, Volkan Seyrantepe
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引用次数: 0

摘要

背景:Tay-Sachs病(TSD)是一种常染色体隐性溶酶体贮积症,其特征是由于编码β-已糖氨酸酶a α-亚基的HEXA基因突变导致GM2神经节苷脂积累,这种积累导致患者显著的神经病理效应和过早死亡。目前还没有有效的治疗方法,但酶替代疗法正在研究中。在我们之前的工作中,我们证明了人重组溶酶体β-己糖氨酸酶A (rhHex-A)的内化和有效性,该酶A由甲基营养酵母毕赤酵母产生,在降低成纤维细胞和患者诱导的多能干细胞(iPSCs)衍生的神经干细胞的脂质和溶酶体质量水平方面。在这项研究中,我们利用来自创伤后应激障碍患者的成纤维细胞和神经胶质细胞以及相关小鼠模型进一步评估了rhHex-A预防GM2积累的潜力。方法:用100 nM rhHexA处理小鼠模型和TSD患者的成纤维细胞和神经胶质细胞系72 h,处理后用抗GM2(靶向GM2神经节苷脂;KM966)和抗lamp1(溶酶体相关膜蛋白1)共定位染色,并用50 nM LysoTracker Red ddn -99孵育,标记溶酶体。此外,我们还分析了GM2AP和HEXB的表达,以评估rhHex-A处理是否会影响GM2神经节苷脂降解相关酶的水平。结果:LysoTracker的免疫荧光染色和GM2和Lamp1的共定位研究表明,溶酶体质量和GM2水平降低。值得注意的是,rhHex-A处理还影响了参与GM2神经节苷脂代谢的HEXB基因的表达,突出了代谢途径中潜在的调节相互作用。结论:本文报道酵母中产生的rhHex-A能够有效降解GM2神经节苷脂,并挽救TSD细胞中溶酶体的积累。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Yeast-Produced Human Recombinant Lysosomal β-Hexosaminidase Efficiently Rescues GM2 Ganglioside Accumulation in Tay-Sachs Disease.

Background: Tay-Sachs disease (TSD) is an autosomal recessive lysosomal storage disorder characterized by the accumulation of GM2 ganglioside due to mutations in the HEXA gene, which encodes the α-subunit of β-Hexosaminidase A. This accumulation leads to significant neuropathological effects and premature death in affected individuals. No effective treatments exist, but enzyme replacement therapies are under investigation. In our previous work, we demonstrated the internalization and efficacy of human recombinant lysosomal β-hexosaminidase A (rhHex-A), produced in the methylotrophic yeast Pichia pastoris, in reducing lipids and lysosomal mass levels in fibroblasts and neural stem cells derived from patient-induced pluripotent stem cells (iPSCs). In this study, we further evaluated the potential of rhHex-A to prevent GM2 accumulation using fibroblast and neuroglia cells from a TSD patient alongside a relevant mouse model. Methods: Fibroblasts and neuroglial cell lines derived from a murine model and TSD patients were treated with 100 nM rhHexA for 72 h. After treatment, cells were stained by anti-GM2 (targeting GM2 ganglioside; KM966) and anti-LAMP1 (lysosomal-associated membrane protein 1) colocalization staining and incubated with 50 nM LysoTracker Red DND-99 to label lysosomes. In addition, GM2AP and HEXB expression were analyzed to assess whether rhHex-A treatment affected the levels of enzymes involved in GM2 ganglioside degradation. Results: Immunofluorescence staining for LysoTracker and colocalization studies of GM2 and Lamp1 indicated reduced lysosomal mass and GM2 levels. Notably, rhHex-A treatment also affected the expression of the HEXB gene, which is involved in GM2 ganglioside metabolism, highlighting a potential regulatory interaction within the metabolic pathway. Conclusions: Here, we report that rhHex-A produced in yeast can efficiently degrade GM2 ganglioside and rescue lysosomal accumulation in TSD cells.

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来源期刊
Journal of Personalized Medicine
Journal of Personalized Medicine Medicine-Medicine (miscellaneous)
CiteScore
4.10
自引率
0.00%
发文量
1878
审稿时长
11 weeks
期刊介绍: Journal of Personalized Medicine (JPM; ISSN 2075-4426) is an international, open access journal aimed at bringing all aspects of personalized medicine to one platform. JPM publishes cutting edge, innovative preclinical and translational scientific research and technologies related to personalized medicine (e.g., pharmacogenomics/proteomics, systems biology). JPM recognizes that personalized medicine—the assessment of genetic, environmental and host factors that cause variability of individuals—is a challenging, transdisciplinary topic that requires discussions from a range of experts. For a comprehensive perspective of personalized medicine, JPM aims to integrate expertise from the molecular and translational sciences, therapeutics and diagnostics, as well as discussions of regulatory, social, ethical and policy aspects. We provide a forum to bring together academic and clinical researchers, biotechnology, diagnostic and pharmaceutical companies, health professionals, regulatory and ethical experts, and government and regulatory authorities.
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