Gene network analysis to identify hub genes and biological pathways related to mastitis in dairy cows based on bioinformatics analysis

Document Type : Research Paper

Authors

1 *Corresponding author, Associate Professor, Department of Animal Science, Faculty of Agriculture, University of Jiroft, Jiroft, Iran

2 PhD of animal breeding, Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran

3 PhD Student, Department of Animal Science, Faculty of Agriculture, University of Zabul, Zabul, Iran

Abstract

Objective
Mastitis, a significant infectious ailment affecting dairy cows during both dry and lactation periods, has substantial impacts on animal health, economic profitability, and the dairy industry each year. This study seeks to utilize bioinformatics analysis not only to identify hub genes and biological pathways linked to mastitis in dairy cows but also to gain a deeper understanding of its implications.
Materials and methods
21 microarray data samples were obtained from the GEO database (accession number GSE24217), comprising 9 healthy cow tissue samples and 12 E. coli infected tissue samples. These samples were assessed using GEO2R. Differentially expressed genes were identified based on two criteria: adjP-value > 0.05 and |LogFC| ≥ 1. Hub genes were identified using the CytoHubba plugin in Cytoscape software and topological analysis methods (DMNC, MCC, MNC, DEGREE). Signaling pathways were identified using the STRING tool based on an FDR > 0.05 index.
Results
Out of 631 differentially expressed genes, 136 had low expression, and 495 had high expression in infected breast tissue compared to healthy ones. Among these, 205 genes were co-expressed in the network. 12 hub genes, including CCL19, ALB, GAPDH, PTPRC, ICAM1, IL6, IL1B, IL18, CXCL8, CCL20, CXCL16, and CCL3, were identified. The analysis of biological pathways revealed that the studied genes are involved in 66 biological pathways, with NOD-like receptor signaling pathway, TNF signaling pathway, and IL-17 signaling pathway playing important functional roles in dairy cow mastitis disease.
Conclusions
The discovered genes and pathways have the potential to enhance our knowledge of the complex molecular mechanisms involved in mastitis development. These discoveries could shape a thorough reform initiative focused on preventing, diagnosing, and treating mastitis. These insights are expected to pave the way for the creation of precise and dependable biomarkers for early detection and prevention of bovine E. coli mastitis, ultimately facilitating personalized treatment approaches.

Keywords


جعفری احمدآبادی سید علی اصغر، عسکری­همت حشمت­اله، محمدآبادی محمدرضا (1402) تاثیر شاهدانه بر بیان ژن DLK1 در بافت‌ قلب بره‌های کرمانی. مجله بیوتکنولوژی کشاورزی، 15(1)، 217-234.
شکری سمیرا، خضری امین، محمدآبادی محمدرضا، خیرالدین حمید (1402) بررسی بیان ژن MYH7  در بافت‌های ران، دست و راسته بره‌های پرواری نژاد کرمانی. مجله بیوتکنولوژی کشاورزی، 15(2)، 217-236.
وجدی حکم‌آباد صمد، علیجانی صادق، دقیق‌کیا حسین و همکاران (1394) مطالعه بیوانفورماتیکی بیماری ورم پستان گاو ایجاد شده توسط لیپوپلی ساکارید اشرشیاکلی با استفاده از داده‌های ریزآرایه. مجله کومش 17(۱)، ۲۱۴-۲۲۳.
References
Asselstine VJ, Medrano F, Cánovas A (2022) Identification of Novel Alternative Splicing Associated with Mastitis Disease in Holstein Dairy Cows Using Large Gap Read Mapping. BMC Genomics 23(1), 1–15.
Bakhtiarizadeh MR, Mirzaei S, Norouzi M, et al.) 2020) Identification of Gene Modules and Hub Genes Involved in Mastitis Development Using a Systems Biology Approach. Front Genet 11, e722.
Barazandeh A, Mohammadabadi MR, Ghaderi Zefrehei M, et al. (2016a) Predicting CpG Islands and Their Relationship with Genomic Feature in Cattle by Hidden Markov Model Algorithm. Iran J Appl Anim Sci 6 (3), 571-579.
Barazandeh A, Mohammadabadi MR, Ghaderi Zefrehei M, et al. (2016b) Genome-wide analysis of CpG islands in some livestock genomes and their relationship with genomic features. Czech J Anim Sci 61, 487.
Barlow J (2011) Mastitis Therapy and Antimicrobial Susceptibility: A Multispecies Review with a Focus on Antibiotic Treatment of Mastitis in Dairy Cattle. J Mammary Gland Biol Neoplasia 16, 383–407.
Bordbar F, Mohammadabadi M, Jensen J, et al. (2022) Identification of candidate genes regulating carcass depth and hind leg circumference in simmental beef cattle using Illumina Bovine Beadchip and next-generation sequencing. Animals (Basel) 12 (9), e1103.
Buitenhuis B, Røntved CM, Edwards SM, et al. (2011) In Depth Analysis of Genes and Pathways of the Mammary Gland Involved in the Pathogenesis of Bovine Escherichia Coli-Mastitis. BMC Genomics 12, 1–10.
Chen X, Cheng Z, Zhang S, et al. (2015) Combining Genome Wide Association Studies and Differential Gene Expression Data Analyses Identifies Candidate Genes Affecting Mastitis Caused by Two Different Pathogens in the Dairy Cow. Open J Anim Sci 5(04), 358.
Chin CH, Chen SH, Wu HH, et al. (2014) CytoHubba: Identifying Hub Objects and Sub-Networks from Complex Interactome. BMC Syst Biol 8(4), 1–7.
Ciftci AB, Yemez K, Polat S, Yazıcıoğlu M (2022) Risk Factors and the Role of the Albumin-to-Globulin Ratio in Predicting Recurrence among Patients with Idiopathic Granulomatous Mastitis. J Inflamm Res 5401–12.
Darang E, Pezeshkian Z, Ziaeddin Mirhoseini S, Ghovvati S (2023). Identification of Key Genes and Potential Pathways Associated with Mastitis Induced by E. Coli. Biochem Genet 61(1), 202–20.
Dong Y, Yu JJ, Shibahara Y, Lu H, He HY, et al. (2014) Intercellular Adhesion Molecule 1/2 and E-Selectin in Plasma Cell Mastitis: Immunohistochemical Study of 35 Cases. Hum Pathol 45(3), 606–10.
Ghahramani N, Shodja J, Rafat SA, Panahi B, Hasanpur K (2021) Integrative Systems Biology Analysis Elucidates Mastitis Disease Underlying Functional Modules in Dairy Cattle. Front Genet 12, 712306.
Goji N, Potter AA, Perez-Casal J (2004) Characterization of Two Proteins of Staphylococcus Aureus Isolated from Bovine Clinical Mastitis with Homology to Glyceraldehyde-3-Phosphate Dehydrogenase. Vet Microbiol 99(3–4), 269–79.
Hamed AE, Mohammed A, Kamel E (2020) Incidence and Economics of Clinical Mastitis of Holstein Friesian Dairy Cows under Egyptian Condition. Benha Vet Med J 39(1), 119–24.
Jafari Ahmadabadi SAA, Askari-Hemmat H, Mohammadabadi M, et al. (2023) The effect of Cannabis seed on DLK1 gene expression in heart tissue of Kermani lambs. J Agric Biotechnol 15 (1), 217-234 (In Persian).
Jaiswal S, Jagannadham J, Kumari J, Iquebal MA, Gurjar AK, Nayan V, Angadi UB, Kumar S, Kumar R, Datta TK (2021) Genome Wide Prediction, Mapping and Development of Genomic Resources of Mastitis Associated Genes in Water Buffalo. Front Vet Sci 8, 593871.
Ju Z, Jiang Q, Wang J, Wang X, Yang C, Sun Y, Zhang Y, Wang C, Gao Y, Wei X (2020) Genome-Wide Methylation and Transcriptome of Blood Neutrophils Reveal the Roles of DNA Methylation in Affecting Transcription of Protein-Coding Genes and MiRNAs in E. Coli-Infected Mastitis Cows. BMC Genomics 21, 1–14.
Khan MZ, Khan A, Xiao J, Ma Y, Ma J, Gao J, Cao Z (2020) Role of the JAK-STAT Pathway in Bovine Mastitis and Milk Production. Animals (Basel) 10(11), 2107.
Kobayashi K, Matsunaga K, Tsugami Y, Wakasa H, Nishimura T (2021) IL-1β Is a Key Inflammatory Cytokine That Weakens Lactation-Specific Tight Junctions of Mammary Epithelial Cells. Exp Cell Res 409(2), 112938.
Korbecki J, Bajdak-Rusinek K, Kupnicka P, Kapczuk P, Chlubek D, Baranowska-Bosiacka I (2021) The Role of CXCL16 in the Pathogenesis of Cancer and Other Diseases. Int J Mol Sci 22(7), 3490.
Li L, Chen X, Chen Z (2019) Identification of Key Candidate Genes in Dairy Cow in Response to Escherichia Coli Mastitis by Bioinformatical Analysis. Front Genet 10, 1251.
Luoreng Z, Wang XP, Mei CG, Zan LS (2018) Expression Profiling of Peripheral Blood MiRNA Using RNAseq Technology in Dairy Cows with Escherichia Coli-Induced Mastitis. Sci Rep 8(1), 12693.
Masoudzadeh, S.H., Mohammadabadi, M.R., Khezri, A., et al. (2020) Dlk1 gene expression in different Tissues of lamb. Iran J Appl Anim Sci 10, 669-677.
Mohamadipoor L, Mohammadabadi M, Amiri Z, et al. (2021) Signature selection analysis reveals candidate genes associated with production traits in Iranian sheep breeds. BMC Vet Res 17 (1), 1-9.
Mohammadabadi M, Masoudzadeh SH, Khezri A, et al. (2021) Fennel (Foeniculum vulgare) seed powder increases Delta-Like Non-Canonical Notch Ligand 1 gene expression in testis, liver, and humeral muscle tissues of growing lambs. Heliyon 7 (12), e08542.
Mohammadinejad F, Mohammadabadi M, Roudbari Z, Sadkowski T (2022) Identification of Key Genes and Biological Pathways Associated with Skeletal Muscle Maturation and Hypertrophy in Bos taurus, Ovis aries, and Sus scrofa. Animals (Basel) 12 (24), 3471.
Roussel P, Cunha P, Porcherie A, Petzl W, Gilbert FB, Riollet C, Zerbe H, Rainard P, Germon P (2015) Investigating the Contribution of IL-17A and IL-17F to the Host Response during Escherichia Coli Mastitis. Vet Res 46. 1–14.
Ruegg PL (2017) A 100-Year Review: Mastitis Detection, Management, and Prevention. J Dairy Sci 100(12), 10381–97.
Safaei SMH, Dadpasand M, Mohammadabadi M, et al. (2022) An Origanum majorana Leaf Diet Influences Myogenin Gene Expression, Performance, and Carcass Characteristics in Lambs. Animals (Basel) 13 (1), e14.
Schmelcher M, Powell AM, Camp MJ, Pohl CS, Donovan DM (2015) Synergistic Streptococcal Phage ΛSA2 and B30 Endolysins Kill Streptococci in Cow Milk and in a Mouse Model of Mastitis. Appl Microbiol Biotechnol 99, 8475–86.
Shahsavari M, Mohammadabadi M, Khezri A, et al. (2022) Effect of Fennel (Foeniculum Vulgare) Seed Powder Consumption on Insulin-like Growth Factor 1 Gene Expression in the Liver Tissue of Growing Lambs. Gene Expr 21 (2), 21-26.
Shokri S, Khezri A, Mohammadabadi M, et al. (2023). The expression of MYH7 gene in femur, humeral muscle and back muscle tissues of fattening lambs of the Kermani breed. J Agric Biotechnol 15 (2), 217-236 (In Persian).
Tarca AL, Romero R, Draghici S (2006)  Analysis of Microarray Experiments of Gene Expression Profiling. Am J Obstet Gynecol 195(2), 373–88.
Vajdi Hakamabad S, Alijani HD, Kia H, et al. (2015) Bioinformatics Analysis of E. Coli Causing Mastitis in Holstein Dairy Cattle by Using Microarray Data. Faslnamahi Kumish 17(1), 214-223 (In Persian).
Zheng J, Anjanette D, Watson D, et al. (2006) Genome-Wide Expression Analysis of Lipopolysaccharide-Induced Mastitis in a Mouse Model. Infect Immun 74(3), 1907–15.
Zhu X, Chen M, Wang H, et al. (2021) Clinical Utility of Expanded Non‐invasive Prenatal Screening and Chromosomal Microarray Analysis in High‐risk Pregnancy. Ultrasound Obstet Gynecol 57(3), 459–65.