Genetic and pharmacologic p32-inhibition rescue CHCHD2-linked Parkinson's disease phenotypes in vivo and in cell models.

IF 9 2区 医学 Q1 CELL BIOLOGY Journal of Biomedical Science Pub Date : 2024-02-23 DOI:10.1186/s12929-024-01010-z
Murni Tio, Rujing Wen, Cai Ning Choo, Jian Bin Tan, Aaron Chua, Bin Xiao, Jeyapriya Rajameenakshi Sundaram, Christine Hui Shan Chan, Eng-King Tan
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Abstract

Background: Mutations in CHCHD2 have been linked to Parkinson's disease, however, their exact pathophysiologic roles are unclear. The p32 protein has been suggested to interact with CHCHD2, however, the physiological functions of such interaction in the context of PD have not been clarified.

Methods: Interaction between CHCHD2 and p32 was confirmed by co-immunoprecipitation experiments. We studied the effect of p32-knockdown in the transgenic Drosophila and Hela cells expressing the wild type and the pathogenic variants of hCHCHD2. We further investigated the rescue ability of a custom generated p32-inhibitor in these models as well as in the human fibroblast derived neural precursor cells and the dopaminergic neurons harboring hCHCHD2-Arg145Gln.

Results: Our results showed that wildtype and mutant hCHCHD2 could bind to p32 in vitro, supported by in vivo interaction between human CHCHD2 and Drosophila p32. Knockdown of p32 reduced mutant hCHCHD2 levels in Drosophila and in vitro. In Drosophila hCHCHD2 models, inhibition of p32 through genetic knockdown and pharmacological treatment using a customized p32-inhibitor restored dopaminergic neuron numbers and improved mitochondrial morphology. These were correlated with improved locomotor function, reduced oxidative stress and decreased mortality. Consistently, Hela cells expressing mutant hCHCHD2 showed improved mitochondrial morphology and function after treatment with the p32-inhibitor. As compared to the isogenic control cells, large percentage of the mutant neural precursor cells and dopaminergic neurons harboring hCHCHD2-Arg145Gln contained fragmented mitochondria which was accompanied by lower ATP production and cell viability. The NPCs harboring hCHCHD2-Arg145Gln also had a marked increase in α-synuclein expression. The p32-inhibitor was able to ameliorate the mitochondrial fragmentation, restored ATP levels, increased cell viability and reduced α-synuclein level in these cells.

Conclusions: Our study identified p32 as a modulator of CHCHD2, possibly exerting its effects by reducing the toxic mutant hCHCHD2 expression and/or mitigating the downstream effects. Inhibition of the p32 pathway can be a potential therapeutic intervention for CHCHD2-linked PD and diseases involving mitochondrial dysfunction.

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基因和药物 p32 抑制可在体内和细胞模型中挽救与 CHCHD2 相关的帕金森病表型。
背景:CHCHD2突变与帕金森病有关,但其确切的病理生理作用尚不清楚。有人认为 p32 蛋白与 CHCHD2 相互作用,但这种作用在帕金森病中的生理功能尚未明确:方法:通过共免疫沉淀实验证实了CHCHD2和p32之间的相互作用。我们研究了在表达hCHCHD2野生型和致病变体的转基因果蝇和Hela细胞中敲除p32的效果。我们还进一步研究了在这些模型以及人成纤维细胞衍生的神经前体细胞和携带 hCHCHD2-Arg145Gln 的多巴胺能神经元中定制生成的 p32 抑制剂的拯救能力:结果:我们的研究结果表明,野生型和突变型hCHCHD2可在体外与p32结合,人CHCHD2与果蝇p32的体内相互作用也证实了这一点。p32被敲除后,果蝇和体外的突变型hCHCHD2水平降低。在hCHCHD2果蝇模型中,通过基因敲除和使用定制的p32抑制剂进行药物治疗来抑制p32,可以恢复多巴胺能神经元的数量并改善线粒体形态。这与运动功能改善、氧化应激减少和死亡率降低相关。同样,表达突变型 hCHCHD2 的 Hela 细胞在使用 p32 抑制剂处理后,线粒体形态和功能也得到了改善。与同源对照细胞相比,携带 hCHCHD2-Arg145Gln 的突变神经前体细胞和多巴胺能神经元中有很大比例的线粒体破碎,同时 ATP 产量和细胞存活率也较低。携带 hCHCHD2-Arg145Gln 的神经元还明显增加了 α-突触核蛋白的表达。p32抑制剂能够改善这些细胞的线粒体破碎、恢复ATP水平、提高细胞活力并降低α-突触核蛋白水平:我们的研究发现 p32 是 CHCHD2 的调节剂,可能通过减少毒性突变体 hCHCHD2 的表达和/或减轻下游效应来发挥其作用。抑制p32通路可能是治疗与CHCHD2相关的帕金森病和涉及线粒体功能障碍的疾病的一种潜在干预措施。
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来源期刊
Journal of Biomedical Science
Journal of Biomedical Science 医学-医学:研究与实验
CiteScore
18.50
自引率
0.90%
发文量
95
审稿时长
1 months
期刊介绍: The Journal of Biomedical Science is an open access, peer-reviewed journal that focuses on fundamental and molecular aspects of basic medical sciences. It emphasizes molecular studies of biomedical problems and mechanisms. The National Science and Technology Council (NSTC), Taiwan supports the journal and covers the publication costs for accepted articles. The journal aims to provide an international platform for interdisciplinary discussions and contribute to the advancement of medicine. It benefits both readers and authors by accelerating the dissemination of research information and providing maximum access to scholarly communication. All articles published in the Journal of Biomedical Science are included in various databases such as Biological Abstracts, BIOSIS, CABI, CAS, Citebase, Current contents, DOAJ, Embase, EmBiology, and Global Health, among others.
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