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. 2021 Jul;22(1):784.
doi: 10.3892/etm.2021.10216. Epub 2021 May 19.

Bioinformatics identification of hub genes and signaling pathways regulated by intravenous immunoglobulin treatment in acute Kawasaki disease

Hongbiao Huang  1 , Lei Xu  1 , Yueyue Ding  1 , Jie Qin  1 , Chengcheng Huang  1 , Xuan Li  1 , Yunjia Tang  1 , Guanghui Qian  1 , Haitao Lv  1
Affiliations

Bioinformatics identification of hub genes and signaling pathways regulated by intravenous immunoglobulin treatment in acute Kawasaki disease

Hongbiao Huang et al. Exp Ther Med. 2021 Jul.

Abstract

Kawasaki disease (KD) is an acute, self-limiting form of vasculitis commonly encountered in infants and young children. Intravenous immunoglobulin (IVIG) is the primary drug used for the treatment of KD, which may significantly reduce the occurrence of coronary artery lesions. However, the specific molecular profile changes of KD caused by IVIG treatment have remained elusive and require further research. The present study was designed to identify key genes, pathways and immune cells affected by IVIG treatment using multiple bioinformatics analysis methods. The results suggested that myeloid cells and neutrophils were affected by IVIG treatment. Kyoto Encyclopedia of Genes and Genomes pathway analysis identified that hematopoietic cell lineages and osteoclast differentiation may have an important role in the mechanism of action of IVIG treatment. Immune cell analysis indicated that the levels of monocytes, M1 macrophages, neutrophils and platelets were markedly changed in patients with KD after vs. prior to IVIG treatment. The key upregulated genes, including ZW10 interacting kinetochore protein, GINS complex subunit 1 and microRNA-30b-3p in whole blood cells of patients with KD following treatment with IVIG indicated that these IVIG-targeted molecules may have important roles in KD. In addition, these genes were further examined by literature review and indicated to be involved in cell proliferation, apoptosis and virus-related immune response in patients with KD. Therefore, the present results may provide novel insight into the mechanisms of action of IVIG treatment for KD.

Keywords: Kawasaki disease; bioinformatics analysis; intravenous immunoglobulin; mechanism of action.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Volcano plot map of (A) differentially expressed genes and (B) differentially expressed proteins. Red dots, upregulated genes/proteins; green dots, downregulated genes/proteins.
Figure 2
Figure 2
Venn diagrams illustrating the number of GO pathways in DEGs and DEPs. (A) The GO pathways of DEPs are represented by a blue circle. (B) The GO pathways of DEGs are represented by a red circle. The intersection in dark red represents 38 pathways the two groups had in common. GO, gene ontology; DEGs, differentially expressed genes; DEPs, differentially expressed proteins; FC, fold change; P.adj, adjusted P-value.
Figure 3
Figure 3
GO enrichment analysis of DEGs and DEPs. For (A) DEGs and (B) DEPs, the top 10 pathways in each GO category (MF, CC and BP) are presented in bubble charts. If the analysis results yielded >10 pathways enriched in this category, the top 10 pathways were listed according to the FDR. (C) Venn diagram indicating the GO pathways for the DEGs and DEPs. The top 10 GO pathways for the DEPs in each GO category were determined and represented by a blue circle, while those for the DEGs were represented by a red circle. GO, gene ontology; MF, molecular function; CC, cellular component; BP, biological process; FDR, false discovery rate; DEGs, differentially expressed genes; DEPs, differentially expressed proteins.
Figure 4
Figure 4
KEGG enrichment analysis of DPGs and DEGs. (A) KEGG enrichment analysis results of (A) DEGs and (B) DPGs presented in bubble charts. (C) Venn diagrams illustrating the number of KEGG pathways for DEGs and DEPs. KEGG pathways calculated for DEPs are indicated by a blue circle and those for DEGs are represented by a red circle. The intersection in dark red represents two pathways that were common between DEGs and DEPs. KEGG, Kyoto Encyclopedia of Genes and Genomes; DEGs, differentially expressed genes; DEGs, differentially expressed genes; DEPs, differentially expressed proteins; FDR, false discovery rate.
Figure 5
Figure 5
Comparison of the xCell scores of 64 cell types between pre- and post-IVIG treatment whole-blood cell samples of patients with Kawasaki disease in the GES48498 dataset. Violin charts displaying significantly different lymphoid cells. *P<0.05. IVIG, intravenous immunoglobulin; MEP, megakaryocyte/erythroid progenitor cells; Th1, type I T-helper.
Figure 6
Figure 6
Protein-protein interaction network of differentially expressed genes between the before- and after-intravenous immunoglobulin Kawasaki disease samples. Yellow and elliptical nodes, hub genes; red and elliptical nodes, upregulated genes; green and diamond nodes, downregulated genes.
Figure 7
Figure 7
Highest-scoring gene cluster obtained from the protein-protein interaction network with 17 nodes and 126 edges.
Figure 8
Figure 8
mRNA-miRNA network of hub genes. Yellow and rectangular nodes represent the miRNAs targeting ≥3 mRNAs. Red and elliptical nodes represent mRNAs that interact with miRNAs. Green and diamond nodes represent miRNAs connected to <3 mRNAs. miRNA/miR, microRNA.
Figure 9
Figure 9
RT-qPCR results. (A) The relative expression levels of ZWINT (P=0.00446) and GINS1 (P=0.00029) in a patient with KD prior to IVIG treatment were higher than those after IVIG treatment. (B) The relative levels of miR-30b-3p expression (P=0.00012) were lower prior to IVIG treatment compared with those thereafter. ***P<0.01. Statistical analysis was performed using Wilcoxon's signed-rank test. RT-qPCR, reverse transcription-quantitative PCR; KD, Kawasaki disease; IVIG, intravenous immunoglobulin; ZWINT, ZW10 interacting kinetochore protein; GINS1, GINS complex subunit 1; TOP2A, topoisomerase II alpha; RRM2, ribonucleotide reductase regulatory subunit M2; TK1, thymidine kinase 1.
Figure 10
Figure 10
Results of cell experiment. (A) Relative expression levels of ICAM1 (P=0.046) in HCAECs stimulated with LPS prior to IVIG treatment were higher than those after IVIG treatment. (B) The relative levels of ZWINT (P=0.049) and GINS1 (P=0.049) in HCAECs stimulated with LPS prior to IVIG treatment were lower than those after IVIG treatment. miR-30b-3p expression (P=0.043) in HCAECs stimulated with LPS prior to IVIG treatment was higher as compared with that after treatment. (C) HCAEC apoptosis was observed following treatment with 1 µg/ml LPS for 4 h and cell proliferation was accelerated following treatment with 25 mg/ml IVIG for 18 h, compared with the control group. Magnification, x400. *P<0.05. Statistical analysis was performed using the Wilcoxon rank-sum test. ICAM1, intercellular adhesion molecule 1; HCAECs, human coronary artery endothelial cells; LPS, lipopolysaccharide; IVIG, intravenous immunoglobulin; ZWINT, ZW10 interacting kinetochore protein; GINS1, GINS complex subunit 1.
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References

    1. de Graeff N, Groot N, Ozen S, Eleftheriou D, Avcin T, Bader-Meunier B, Dolezalova P, Feldman BM, Kone-Paut I, Lahdenne P, et al. European consensus-based recommendations for the diagnosis and treatment of Kawasaki disease - the SHARE initiative. Rheumatology (Oxford) 2019;58:672–682. doi: 10.1093/rheumatology/key344. - DOI - PubMed
    1. Lo MS, Newburger JW. Role of intravenous immunoglobulin in the treatment of Kawasaki disease. Int J Rheum Dis. 2018;21:64–69. doi: 10.1111/1756-185X.13220. - DOI - PubMed
    1. Furusho K, Kamiya T, Nakano H, Kiyosawa N, Shinomiya K, Hayashidera T, Tamura T, Hirose O, Manabe Y, Yokoyama T, et al. High-dose intravenous gammaglobulin for Kawasaki disease. Lancet. 1984;2:1055–1058. doi: 10.1016/s0140-6736(84)91504-6. - DOI - PubMed
    1. Newburger JW, Takahashi M, Beiser AS, Burns JC, Bastian J, Chung KJ, Colan SD, Duffy CE, Fulton DR, Glode MP, et al. A single intravenous infusion of gamma globulin as compared with four infusions in the treatment of acute Kawasaki syndrome. N Engl J Med. 1991;324:1633–1639. doi: 10.1056/NEJM199106063242305. - DOI - PubMed
    1. Sosa T, Brower L, Divanovic A. Diagnosis and Management of Kawasaki Disease. JAMA Pediatr. 2019;173:278–279. doi: 10.1001/jamapediatrics.2018.3307. - DOI - PubMed

Grants and funding

Funding: The present study was supported by grants from the National Natural Science Foundation of China (grant nos. 81971477, 81870365, 82070512 and 81970436), Jiangsu Provincial Medical Young Talents (grant no. QNRC2016756), the Applied Foundational Research of Medical and Health Care of Suzhou City (grant no. SYS2019086), Suzhou Medical Key Discipline - Pediatric Cardiology (grant no. Szxk201507), Key Medical Talents in Jiangsu Province (grant no. ZDRCA2016049), Second cycle key subjects of maternal and child health in Jiangsu Province (grant no. FXK201740) and Introduction Project of Clinical Medical Expert Team in Suzhou City (grant no. SZYJTD201805).