Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 41944, Korea.
BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Sciences, School of Medicine, Kyungpook National University, Daegu 412944, Korea.
Cells. 2021 Mar 18;10(3):676. doi: 10.3390/cells10030676.
Transactive response DNA-binding protein 43 (TDP-43) is a ubiquitously expressed DNA/RNA-binding protein linked to amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). TDP-43 has been implicated in numerous aspects of the mRNA life cycle, as well as in cell toxicity and neuroinflammation. In this study, we used the toxicity of the TDP-43 expression in as an assay to identify genetic interactions. Specifically, we transformed human cDNAs of wild-type or disease-associated mutants ( and ) en masse into 4653 homozygous diploid yeast deletion mutants and then used next-generation sequencing readouts of growth to identify yeast toxicity modifiers. Genetic interaction analysis provided a global view of TDP-43 pathways, some of which are known to be involved in cellular metabolic processes. Selected putative loci with the potential of genetic interactions with were assessed for associations with neurotoxicity and inflammatory activation of astrocytes. The pharmacological inhibition of succinate dehydrogenase flavoprotein subunit A (SDHA) and voltage-dependent anion-selective channel 3 (VDAC3) suppressed TDP-43-induced expression of proinflammatory cytokines in astrocytes, indicating the critical roles played by SDHA and VDAC3 in TDP-43 pathways during inflammatory activation of astrocytes and neuroinflammation. Thus, the findings of our genetic interaction screen provide a global landscape of TDP-43 pathways and may help improve our understanding of the roles of glia and neuroinflammation in ALS and FTD pathogenesis.
转录激活反应 DNA 结合蛋白 43（TDP-43）是一种普遍表达的 DNA/RNA 结合蛋白，与肌萎缩侧索硬化症（ALS）和额颞叶痴呆（FTD）有关。TDP-43 涉及 mRNA 生命周期的多个方面，以及细胞毒性和神经炎症。在这项研究中，我们使用 TDP-43 在 中的表达毒性作为测定方法来识别遗传相互作用。具体来说，我们将野生型或与疾病相关的突变体（和）的人 cDNA 大规模转化为 4653 个纯合二倍体酵母缺失突变体，然后使用下一代测序读取物的生长来识别酵母毒性修饰剂。遗传相互作用分析提供了 TDP-43 途径的全局视图，其中一些已知与细胞代谢过程有关。对具有与 潜在遗传相互作用的选定假定基因座进行了神经毒性和星形胶质细胞炎症激活的评估。琥珀酸脱氢酶黄素蛋白亚单位 A（SDHA）和电压依赖性阴离子选择通道 3（VDAC3）的药理学抑制抑制了 TDP-43 诱导的星形胶质细胞中促炎细胞因子的表达，表明 SDHA 和 VDAC3 在星形胶质细胞炎症激活和神经炎症过程中在 TDP-43 途径中发挥关键作用。因此，我们的遗传相互作用筛选结果提供了 TDP-43 途径的全局景观，并可能有助于我们更好地理解胶质细胞和神经炎症在 ALS 和 FTD 发病机制中的作用。
