Bioinformatics analysis of the sensitization mechanisms and molecular targets of Echinococcus granulosus

doi:10.12140/j.issn.1000-7423.2022.03.006

Abstract

Abstract:

Objective To analyze the sensitization mechanisms and molecular targets of Echinococcus granulosus using bioinformatics approaches. Methods Eighteen BALB/c mice were randomly grouped as infection- unsensitized (CE), infection-sensitized (ANPC), and uninfected controls (CTRL). Mice in the infected unsensitized group were injected with protoscoleces suspension (0.2 ml/mouse), followed with saline (0.2 ml/mice) after 90 days. Mice in the infected sensitized group were injected with protoscoleces suspension (0.2 ml/mice), followed with crude cyst fluid (0.2 ml/mice) 90 days after infection. Mice in the uninfected control group were injected with an equal amount of saline. Splenic tissue from three mice of each group were collected 1 h after injection for RNA extraction and transcriptome sequencing. Genes of differential expression were identified by bioinformatics analysis. Significance analysis of differential expression differences was performed using edgeR (quantified with RNA-Seq by expectation-Maximization RSEM) software. Network interaction analysis was performed using the String database and Cytoscape software with the score set as > 500. The network analysis of the genes was then performed using the Cytoscape software and use cytoHubba to obtain the degree of nodes (degree) and the nodes with degrees over 3 were filtered for display. The analysis was performed using the DAVID online enrichment analysis tool, and the results were visualized using the R language ggplot2. Results A total of 743 genes were differentially expressed in the ANPC group compared to the CTRL group, including 568 upregulated genes and 175 downregulated genes. A total of 554 genes were differentially expressed in the CE group compared to the CTRL group, including 393 upregulated genes and 161 downregulated genes. There are 458 genes specifically expressed in the ANPC group compared to the CTRL group, 269 genes specifically expressed in the CE group compared to the CTRL group and 285 genes co-expressed. GO enrichment of 458 genes, which are unique to the ANPC group, and the CTRL group showed that 147 genes are involved in the biological process, 10 genes are involved in cellular components, and 43 genes are involved in molecular functions. The GO enrichment of shared genes showed that 56 were involved in biological processes, 23 in cellular components, and 25 in molecular functions. The GO enrichment of genes that are unique to the CE group and CTRL group revealed that 51 genes are involved in biological processes, 13 genes are involved in cellular components, and 18 genes are involved in molecular functions. The functions of the ANPC group-specific genes and the CTRL group-specific genes mainly focus on inflammatory response and immune response. The functions of genes expressed in both the ANPC and CTRL groups mainly focus on the neuropeptide signaling pathway and cellular response to interleukin-1. The function of CE group-specific genes and the CTRL group-specific genes mainly focuses on the nucleosome assembly and the positive regulation of ERK1 and ERK2 cascades. The genes expressed explicitly in the ANPC group, and the CTRL group was enriched in 32 pathways, including cytokine-cytokine receptor interaction and the JAK-STAT signalling pathway. The genes commonly expressed in all groups were enriched in 9 pathways, including the phosphatidylinositol 3-kinase/protein kinase B (PI3K-Akt) signaling pathway and complement and coagulation cascades. The CE specific genes and the CTRL group-specific genes are enriched to 12 pathways, including Tyrosine metabolism and Linoleic acid metabolism, and identified key genes through the PPI network. Conclusion The inflammatory response plays an important role in metacestode sensitizing process; high expression of IL-6 promotes the occurrance of inflammation; IL-10 functions as an anti-inflammatory factor confronting the host’s inflammation response; IL-13 may be the key gene in developing immune tolerance after metacestode infection. ERK signaling pathway may play an immunomodulatory role during the growth of metacestodes. Cytokine-cytokine receptor interaction pathway and PI3K-Akt signaling pathway were significantly correlated with the sensitization induced by Einococcus infection.

Key words: Echinococcus, Bioinformatic analysis, IL-6/IL-10, Molecular target

CLC Number:

R532.32

XILIZATI Kulaixi, WANG Chun-sheng, WANG Jia-ling, LI Meng, FANG Zhi-yuan, WANG Si-jia, ZHOU Jing-ru, XIANYIDAN Abulajiang, WU Er-ge-li, YE Jian-rong. Bioinformatics analysis of the sensitization mechanisms and molecular targets of Echinococcus granulosus[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(3): 319-323.

Figures/Tables 1

References 19

[1]	Wen H,, Vuitton L,, Tuxun T, et al. Echinococcosis: advances in the 21st century[J]. Clin Microbiol Rev, 2019, 32(2): e00075-e00018.
[2]	Deplazes P,, Rinaldi L,, Alvarez Rojas CA, et al. Global distribution of alveolar and cystic echinococcosis[J]. Adv Parasitol, 2017, 95: 315-493.
[3]	Pezeshki A,, Aminfar H,, Aminzare M. An analysis of common foodborne parasitic zoonoses in slaughtered sheep and cattle in Tehran, Iran, during 2015—2018[J]. Vet World, 2018, 11(10): 1486-1490.
[4]	Nunnari G,, Pinzone MR,, Gruttadauria S, et al. Hepatic echinococcosis: clinical and therapeutic aspects[J]. World J Gastroenterol, 2012, 18(13): 1448-1458.
[5]	Torgerson PR,, Keller K,, Magnotta M, et al. The global burden of alveolar echinococcosis[J]. PLoS Negl Trop Dis, 2010, 4(6): e722.
[6]	Budke CM,, Deplazes P,, Torgerson PR. Global socioeconomic impact of cystic echinococcosis[J]. Emerg Infect Dis, 2006, 12(2): 296-303.
[7]	Zhong B,, Wang Q. Comprehensive prevention and treatment of echinococcosis, helping Shiqu to achieve prosperity[J]. Mod Prev Med, 2020, 47(24): 4417-4421. (in Chinese)
[7]	( 钟波,, 王谦. 综合防治棘球蚴病, 助力石渠奔小康[J]. 现代预防医学, 2020, 47(24): 4417-4421.)
[8]	Ye JR,, Zhang Q,, Xuan Y, et al. Factors associated with echinococcosis-induced perioperative anaphylactic shock[J]. Korean J Parasitol, 2016, 54(6): 769-775.
[9]	Díaz á. Immunology of cystic echinococcosis (hydatid disease)[J]. Br Med Bull, 2017, 124(1): 121-133.
[10]	Yali YS,, Zheng H,, Yu XD, et al. perimental study on the effect of allergic reaction caused by cystic hydatid disease on interleukin-17 and antibody expression[J]. Int J Anestl Res, 2020, 41(9): 833-837. (in Chinese)
[10]	( 亚力·亚森,, 郑宏,, 于晓东, 等. 囊型包虫病所致过敏反应对白细胞介素-17及抗体表达影响的实验研究[J]. 国际麻醉学与复苏杂志, 2020, 41(9): 833-837.)
[11]	Ye JR. Identification of risk factors of anaphylactic shock caused by echinococcosis and study on differential gene expression in lung tissue[D]. Xinjiang Medical University, 2016: 29-30. (in Chinese)
[11]	( 叶建荣. 包虫病所致过敏性休克危险因素的识别和肺组织差异基因表达的研究[D]. 乌鲁木齐: 新疆医科大学, 2016: 29-30.)
[12]	Zhang WR,, Lin XF,, Li X, et al. Transcriptional identification of potential biomarkers of lung adenocarcinoma[J]. J Shanghai Jiao Tong Univ Med Sci, 2020, 40(12): 1598-1606. (in Chinese)
[12]	张伟然,, 林雪峰,, 李鑫, 等. 转录组分析鉴定肺腺癌潜在的生物标志物[J]. 上海交通大学学报(医学版), 2020, 40(12): 1598-1606.)
[13]	Jiang DB,, Shen JG,, Yang XQ, et al. Effects of interleukin 2, 6 on immunological mechanisms in asth-matic children[J]. Chin J Pediatr, 1994, 32(4): 219-221, 254. (in Chinese)
[13]	( 蒋东波,, 沈际皋,, 杨锡强, 等. 哮喘患儿白细胞介素2, 6的检测及其意义[J]. 中华儿科杂志, 1994, 32(4): 219-221, 254.)
[14]	Fang D,, Zhu J. Molecular switches for regulating the differentiation of inflammatory and IL-10-producing anti-inflammatory T-helper cells[J]. Cell Mol Life Sci, 2020(77): 289-303.
[15]	Jenne L,, Kilwinski J,, Scheffold W, et al. IL-5 expressed by CD4⁺ lymphocytes from Echinococcus multilocularis-infected patients[J]. Clin Exp Immunol, 1997, 109(1): 90-97.
[16]	Borish L,, Aarons A,, Rumbyrt J, et al. Interleukin-10 regulation in normal subjects and patie[J]. J Allergy Clin Immunol, 1996, 97(6): 1288-1296.
[17]	Tamir I,, Stolpa JC,, Helgason CD, et al. The RasGAP-binding protein p62dok is a mediator of inhibitory FcgammaRⅡB signals in B CElls[J]. Immunity, 2000, 12(3): 347-358.
[18]	Hijjawi NS,, Al-Radaideh AM,, Rababah EM, et al. Cystic echinococcosis in Jordan: a review of causative species, previous studies, serological and radiological diagnosis[J]. Acta Trop, 2018, 179: 10-16.
[19]	Cucher MA,, Macchiaroli N,, Baldi G, et al. Cystic echinococcosis in South America: systematic review of species and genotypes of Echinococcus granulosus sensu lato in humans and natural domestic hosts[J]. Trop Med Int Health, 2016, 21(2): 166-175.

通路序号 Pathway ID	代谢通路 Metabolism patheway	差异基因数量 No. differentially gene	基因名称 Gene name
mmu05144	疟疾Malaria	7	Il1b/Sele/Tlr2/Csf3/Sdc4/Icam1/Hgf
mmu04060	细胞因子-细胞因子受体相互作用 Cytokine-cytokine receptor interaction	13	Ccl1/Il1a/Il1b/Cxcl1/Cxcl2/Cx3cr1/Csf3/Hgf/Osmr/Inhbb/Pdgfra/Csf2rb2/Csf2rb
mmu04630	Jak-STAT信号通路Jak-STAT signaling pathway	8	Socs3/Socs1/Csf3/Osmr/Cish/Csf2rb2/Csf2rb/Spry4
mmu04010	MAPK信号通路MAPK signaling pathway	11	Gng12/Il1a/Cd14/Il1b/Fgf23/Gadd45b/Map3k8/Hspa1b/Jun/Pdgfra/Gadd45g
mmu04380	破骨细胞分化Osteoclast differentiation	7	Il1a/Il1b/Socs3/Socs1/Fosl2/Junb/Jun
mmu04621	NOD样受体信号通路 NOD-like receptor signaling pathway	5	Il1b/Cxcl1/Cxcl2/Tnfaip3/Mefv
mmu05140	利什曼病Leishmaniasis	5	Il1a/Ptgs2/Il1b/Tlr2/Jun
mmu05323	类风湿性关节炎Rheumatoid arthritis	5	Il1a/Il1b/Tlr2/Icam1/Jun
mmu04620	Toll样受体信号通路 Toll-like receptor signaling pathway	5	Cd14/Il1b/Tlr2/Map3k8/Jun