Abstract
Toxoplasma gondii is a worldwide parasite that can infect almost all kinds of mammals and cause fatal toxoplasmosis in immunocompromised patients. Apoptosis is one of the principal strategies of host cells to clear pathogens and maintain organismal homeostasis, but the mechanism of cell apoptosis induced by T. gondii remains obscure. To explore the apoptosis influenced by T. gondii, Vero cells infected or uninfected with the parasite were subjected to apoptosis detection and subsequent dual RNA sequencing (RNA-seq). Using high-throughput Illumina sequencing and bioinformatics analysis, we found that pro-apoptosis genes such as DNA damage-inducible transcript 3 (DDIT3), growth arrest and DNA damage-inducible α (GADD45A), caspase-3 (CASP3), and high-temperature requirement protease A2 (HtrA2) were upregulated, and anti-apoptosis genes such as poly(adenosine diphosphate (ADP)-ribose) polymerase family member 3 (PARP3), B-cell lymphoma 2 (Bcl-2), and baculoviral inhibitor of apoptosis protein (IAP) repeat containing 5 (BIRC5) were downregulated. Besides, tumor necrosis factor (TNF) receptor-associated factor 1 (TRAF1), TRAF2, TNF receptor superfamily member 10b (TNFRSF10b), disabled homolog 2 (DAB2)-interacting protein (DAB2IP), and inositol 1,4,5-trisphosphate receptor type 3 (ITPR3) were enriched in the upstream of TNF, TNF-related apoptosis-inducing ligand (TRAIL), and endoplasmic reticulum (ER) stress pathways, and TRAIL-receptor 2 (TRAIL-R2) was regarded as an important membrane receptor influenced by T. gondii that had not been previously considered. In conclusion, the T. gondii RH strain could promote and mediate apoptosis through multiple pathways mentioned above in Vero cells. Our findings improve the understanding of the T. gondii infection process through providing new insights into the related cellular apoptosis mechanisms.
摘要
目的
研究并阐明弓形虫感染宿主细胞后影响细胞凋亡及相关信号通路。
创新点
通过转录组研究发现弓形虫感染促进了细胞凋亡, 并发现了包括DDIT3、GADD45A、CASP3及HtrA2在内的多个关键上调基因。
方法
首先通过流式细胞仪对感染及未感染弓形虫的细胞进行凋亡检测, 提取RNA后进行转录组测序, 通过转录组基因上调、下调分析, GO和KEGG通路、关键基因富集、基因网络关系图、RT-PCR验证等手段进行分析。
结论
流式细胞仪检测发现弓形虫感染后能显著诱导细胞发生凋亡, 进一步的转录组分析发现, 与对照组相比, 感染组有1579个基因表达有显著变化, 其中有918个基因上调, 661个基因下调;在富集的54条通路中, 凋亡通路有16个关键基因上调, 11个基因下调, 其中caspase-3是我们发现的关键网络基因。细胞凋亡与细胞周期密切、与免疫及多种疾病有关, 进一步深入研究和探索弓形虫与宿主细胞之间的凋亡机制可为寻找理想药物靶点提供理论基础。
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Acknowledgments
This research was supported by the National Natural Science Foundation of China (Nos. 81802037 and 81871684), the Qingshan Lake United Fund of Zhejiang Province (No. LQY19H190002), the Zhejiang Provincial Natural Science Foundation of China (No. LY22H190003), the Zhejiang Provincial Program for the Cultivation of High-level Innovative Health Talents, and the Basic Scientific Research Funds of Department of Education of Zhejiang Province (Nos. KYZD202104 and KYYB202101), China.
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The datasets supporting the results of this article are available in the National Center for Biotechnology Information (NCBI) Sequence Read Archive (SRA) repository with the accession number PRJNA791458.
Author contributions
Kaige DU, Fei LU, and Xunhui ZHUO performed the experimental research and data analysis. Chengzuo XIE, Haojie DING, Yu SHEN, and Yafan GAO analyzed the data. Kaige DU and Xunhui ZHUO wrote and edited the manuscript. Shaohong LU contributed to the study design and editing of the manuscript. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Kaige DU, Fei LU, Chengzuo XIE, Haojie DING, Yu SHEN, Yafan GAO, Shaohong LU, and Xunhui ZHUO declare that they have no conflict of interest.
This article does not contain any studies with human or animal subjects performed by any of the authors.
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Du, K., Lu, F., Xie, C. et al. Toxoplasma gondii infection induces cell apoptosis via multiple pathways revealed by transcriptome analysis. J. Zhejiang Univ. Sci. B 23, 315–327 (2022). https://doi.org/10.1631/jzus.B2100877
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DOI: https://doi.org/10.1631/jzus.B2100877