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研究旨在通过从转录组水平分析新冠病毒Beta变异株感染C57BL/6小鼠肺部的基因表达变化，以揭示引发轻症肺部损伤的关键宿主因子。首先构建Beta变异株感染C57BL/6小鼠模型，并对感染和未感染第3天（3d）小鼠的肺组织进行病原检测、病理分析以及差异表达基因分析。最后，筛选可能导致肺部损伤的5个炎症反应关键基因进行RT-qPCR验证。研究结果显示Beta变异株能感染C57BL/6小鼠肺部并导致轻度病理损伤。在感染的小鼠中，Beta变异株激活了肺组织的天然免疫应答，主要通路包括Toll样受体信号通路、Ⅰ和Ⅱ型干扰素反应、白细胞趋化反应和细胞因子反应等。通过对重症与轻症感染小鼠的肺组织转录组比较，筛选出与疾病严重程度相关的炎症反应趋化因子，包括Ccl2、Ccl7、Cxcl9、Cxcl10和Cxcl14。通过RT-qPCR验证，基因表达变化趋势与转录组结果一致，表明这些趋化因子过度激活与肺部病理表型损伤程度相关。本研究揭示了Beta变异株感染小鼠肺部激活宿主炎症反应通路关键因子，可能是引发小鼠肺部轻度病理损伤的主要诱因之一，这为预防和治疗新冠病毒感染提供了有利的研究靶点。
Abstract：This study aimed to analyze the gene expression changes in the lungs of mice infected with Beta variant of SARS-CoV-2 at the transcriptomic level, in order to reveal the expression of mild pulmonary damage induced key host factors. First, a mouse model of C57BL/6 infected with Beta variant was established, and pathogen detection, histopathological analysis, and differential gene expression analysis were conducted on the lung tissues of mice infected and uninfected on the third day. Finally, five key genes involved in inflammatory responses that may contribute to lung damage were selected for validation using RT-qPCR. The results demonstrated that Beta variant effectively infected the lungs of C57BL/6 mice and led to mild pathological damage. In infected mice, Beta variant activated the innate immune responses in lung tissues, primarily including pathways such as Toll-like receptor signaling, type I and type II interferon responses, leukocyte chemotaxis, and cytokine responses. By comparing the transcriptomes of lung tissues between severe and mild infection in mice, inflammatory response chemokines associated with disease severity were identified, including Ccl2, Cc17, CxcI9, Cxcl10 and Cxcl14. RT-qPCR validation confirmed that the gene expression changes were consistent with the transcriptomic results, indicating that the excessive activation of these chemokines was associated with the degree of lung pathological damage. In summary, this study revealed that the infection of mice lungs by Beta variant activates key factors of the host inflammatory response pathway, which may be one of the main causes of mild pathological damage in mice lungs. This provides valuable research targets for the prevention and treatment of SARS-CoV-2 infection.
[1] World Health Organization. Coronavirus disease(COVID-19)outbreak situation[EB/OL].(2023-12-13)[2023-12-13]. https：//data. who. int/dashboards/covid19/deaths.
[2] Li J, Jia H, Tian M, et al. SARS-CoV-2 and emerging variants:unmasking structure, function, infection, and immune escape mechanisms[J]. Front Cell Infect Microbiol, 2022, 12:869832. DOI:10.3389/fcimb.2022.869832.
[3] Chen N S, Zhou M, Dong X, et al. Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China:a descriptive study[J]. Lancet, 2020, 395(10223):507-513. DOI:10.1016/S0140-6736(20)30211-7.
[4] Harrison A G, Lin T, Wang P H. Mechanisms of SARS-CoV-2 transmission and pathogenesis[J]. Trends Immunol, 2020, 41(12):1100-1115. DOI:10.1016/j.it.2020.10.004.
[5] Wang D W, Hu B, Hu C, et al. Clinical characteristics of 138 hospitalized patients with 2019 novel coronavirusinfected pneumonia in Wuhan, China[J]. JAMA,2020, 323(11):1061-1069. DOI:10.1001/jama.2020.1585.
[6] Tegally H, Wilkinson E, Giovanetti M, et al. Detection of a SARS-CoV-2 variant of concern in South Africa[J]. Nature, 2021, 592(7854):438-443. DOI:10.1038/s41586-021-03402-9.
[7] Wibmer C K, Ayres F, Hermanus T, et al. SARS-CoV-2 501Y. V2 escapes neutralization by South African COVID-19 donor plasma[J]. Nat Med, 2021, 27(4):622-625. DOI:10.1038/s41591-021-01285-x.
[8] Tarrés-Freixas F, TrinitéB, Pons-Grífols A, et al.Heterogeneous infectivity and pathogenesis of SARSCoV-2 variants Beta, Delta and Omicron in transgenic K18-hACE2 and wildtype mice[J]. Front Microbiol,2022, 13:840757. DOI:10.3389/fmicb.2022.840757.
[9] Radvak P, Kwon H J, Kosikova M, et al. SARS-CoV-2 B. 1.1.7(alpha)and B. 1.351(beta)variants induce pathogenic patterns in K18-hACE2 transgenic mice distinct from early strains[J]. Nat Commun, 2021, 12(1):6559. DOI:10.1038/s41467-021-26803-w.
[10]Blanco-Melo D, Nilsson-Payant B E, Liu W C, et al.Imbalanced host response to SARS-CoV-2 drives development of COVID-19[J]. Cell, 2020, 181(5):1036-1045.e9. DOI:10.1016/j.cell.2020.04.026.
[11]Yang Y, Shen C G, Li J X, et al. Plasma IP-10 and MCP-3 levels are highly associated with disease severity and predict the progression of COVID-19[J]. J Allergy Clin Immunol, 2020, 146(1):119-127. e4. DOI:10.1016/j.jaci.2020.04.027.
[12]Liu J, Li S, Liu J, et al. Longitudinal characteristics of lymphocyte responses and cytokine profiles in the peripheral blood of SARS-CoV-2 infected patients[J].EBioMedicine, 2020, 55:102763. DOI:10.1016/j.ebiom.2020.102763.
[13]Chen X H, Zhao B H, Qu Y M, et al. Detectable serum severe acute respiratory syndrome coronavirus 2 viral load(RNAemia)is closely correlated with drastically elevated interleukin 6 level in critically ill patients with coronavirus disease 2019[J]. Clin Infect Dis, 2020, 71(8):1937-1942. DOI:10.1093/cid/ciaa449.
[14]Winkler E S, Bailey A L, Kafai N M, et al. SARS-CoV-2 infection of human ACE2-transgenic mice causes severe lung inflammation and impaired function[J]. Nat Immunol, 2020, 21(11):1327-1335. DOI:10.1038/s41590-020-0778-2.
[15]Chen Q, Huang X Y, Tian Y, et al. The infection and pathogenicity of SARS-CoV-2 Variant B. 1.351 in hACE2 mice[J]. Virol Sin, 2021, 36(5):1232-1235.DOI:10.1007/s12250-021-00452-1.
[16]Lu R J, Zhao L, Huang B Y, et al. Real-time reverse transcription-polymerase chain reaction assay panel for the detection of severe acute respiratory syndrome coronavirus 2 and its variants[J]. Chin Med J(Engl),2021, 134(17):2048-2053. DOI:10.1097/CM9.0000000000001687.
[17]Roth R, Madhani H D, Garcia J F. Total RNA isolation and quantification of specific RNAs in fission yeast[J].Methods Mol Biol, 2018, 1721:63-72. DOI:10.1007/978-1-4939-7546-4_6.
[18]Mortazavi A, Williams B A, Mccue K, et al. Mapping and quantifying mammalian transcriptomes by RNA-Seq[J]. Nat Methods, 2008, 5(7):621-628. DOI:10.1038/nmeth.1226.
[19]Liao Y, Smyth G K, Shi W. Feature counts:an efficient general purpose program for assigning sequence reads to genomic features[J]. Bioinformatics, 2014, 30(7):923-930. DOI:10.1093/bioinformatics/btt656.
[20]Moreau G B, Burgess S L, Sturek J M, et al.Evaluation of K18-hACE2 mice as a model of SARSCoV-2 infection[J]. Am J Trop Med Hyg, 2020, 103(3):1215-1219. DOI:10.4269/ajtmh.20-0762.
[21]Oladunni F S, Park J G, Pino P A, et al. Lethality of SARS-CoV-2 infection in K18 human angiotensinconverting enzyme 2 transgenic mice[J]. Nat Commun,2020, 11(1):6122. DOI:10.1038/s41467-020-19891-7.
[22]Fernandez E J, Lolis E. Structure, function, and inhibition of chemokines[J]. Annu Rev Pharmacol Toxicol, 2002, 42:469-499. DOI:10.1146/annurev.pharmtox.42.091901.115838.
[23]Singh S, Anshita D, Ravichandiran V. MCP-1:Function, regulation, and involvement in disease[J].Int Immunopharmacol, 2021, 101(Pt B):107598.DOI:10.1016/j.intimp.2021.107598.
[24]Hughes C E, Nibbs R J B. A guide to chemokines and their receptors[J]. FEBS J, 2018, 285(16):2944-2971. DOI:10.1111/febs.14466.
[25]Propper D J, Balkwill F R. Harnessing cytokines and chemokines for cancer therapy[J]. Nat Rev Clin Oncol,2022, 19(4):237-253. DOI:10.1038/s41571-021-00588-9.
[26]Georgakis M K, Bernhagen J, Heitman L H, et al.Targeting the CCL2-CCR2 axis for atheroprotection[J]. Eur Heart J, 2022, 43(19):1799-1808. DOI:10.1093/eurheartj/ehac094.
[27]Cao S, Liu M F, Sehrawat T S, et al. Regulation and functional roles of chemokines in liver diseases[J]. Nat Rev Gastroenterol Hepatol, 2021, 18(9):630-647.DOI:10.1038/s41575-021-00444-2.
[28]Progatzky F, Shapiro M, Chng S H, et al. Regulation of intestinal immunity and tissue repair by enteric glia[J]. Nature, 2021, 599(7883):125-130. DOI:10.1038/s41586-021-04006-z.
[29]Gerard C, Rollins B J. Chemokines and disease[J]. Nat Immunol, 2001, 2(2):108-115. DOI:10.1038/84209.
[30]Charo I F, Ransohoff R M. The many roles of chemokines and chemokine receptors in inflammation[J]. N Engl J Med, 2006, 354(6):610-621. DOI:10.1056/NEJMra052723.
[31]Zhang H X, Yang K, Chen F, et al. Role of the CCL2-CCR2 axis in cardiovascular disease:Pathogenesis and clinical implications[J]. Front Immunol, 2022, 13:975367. DOI:10.3389/fimmu.2022.975367.
[32]Mercer P F, Williams A E, Scotton C J, et al.Proteinase-activated receptor-1, CCL2, and CCL7regulate acute neutrophilic lung inflammation[J]. Am J Respir cell Mol Biol, 2014, 50(1):144-157. DOI:10.1165/rcmb.2013-0142OC.
[33]Bost P, Giladi A, Liu Y, et al. Host-viral infection maps reveal signatures of severe COVID-19 patients[J]. Cell, 2020, 181(7):1475-1488. e12. DOI:10.1016/j.cell.2020.05.006.
[34]Tomita K, Freeman B L, Bronk S F, et al. CXCL10-mediates macrophage, but not other innate immune cellsassociated inflammation in murine nonalcoholic steatohepatitis[J]. Sci Rep, 2016, 6:28786. DOI:10.1038/srep28786.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004477
中图分类号:R373
引用信息:
[1]张高倩,吴长城,黄保英,等.新型冠状病毒Beta变异株感染C57BL/6小鼠肺组织转录组分析及趋化因子实验验证[J].病毒学报,2024,40(02):255-265.DOI:10.13242/j.cnki.bingduxuebao.004477.
基金信息:
国家重点研发计划（项目号：2023YFC3041500）,题目：奥密克戎变异规律分析与防控研究;国家重点研发计划（项目号：2022YFC2304101）,题目：新冠与流感病毒抗原变异监测及通用型多肽疫苗的临床前评价;国家重点研发计划（项目号：2022YFC2303401）,题目：潜在高危新病毒风险识别的新技术体系研究;国家重点研发计划（项目号：2021YFA1201003）,题目：抗病毒纳米药物的体内外功能评价~~
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