نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشجو

2 پردیس ابوریحان، تخصص: ژنتیک و اصلاح نژاد دام/ ژنتیک مولکولی/ بیوانفورماتیک

3 lهیئت علمی

چکیده

ورم پستان یکی از بیماری‌های عفونی غدد پستان است که هزینه‌های زیادی را به صنعت گاو شیری تحمیل می‌کند. مکانیسم تنظیمی این بیماری پیچیده بوده و تحت کنترل ژن‌های تنظیمی مختلفی قرار دارد. در مطالعه حاضر به منظور درک بهتر عناصر تنظیمی درگیر در بیماری ورم‌پستان، نمونه‌های شیر گاوهای سالم و آلوده در طی سری زمانی صفر، ۱۲، ۳۶،۲۴ و ۴۸ ساعت پس از آلودگی جمع‌آوری گردید. داده‌های miRNA-seq از نمونه‌های شیر بدست آمد و با استفاده از روش‌های بیوانفورماتیکی پیشرفته، miRNA‌های جدید، ژن‌های هدف آن‌ها، عملکرد احتمالی آن‌ها، ایزومیرها و همچنین miRNA*‌های جدید شناسایی شد. نتایج منجر به شناسایی نود و دو miRNA جدید شد که تعداد بیست و شش miRNA دارای ژن همولوگ و شصت و شش miRNA فاقد ژن همولوگ در دیگر گونه‌ها بود. بررسی گروه-های کارکردی ژن‌های هدف، مؤید نقش miRNAهای جدید در بسیاری از مکانیسم‌های مقابله با التهاب و آلودگی از جمله، پاسخ به تحریکات داخلی و خارجی، مرگ سلولی و تولید ایمنوگلوبین است. همچنین علاوه بر این، صد و سی و پنجmiRNA* جدید نیز شناسایی شد. در مطالعه حاضر ۴۹۳ ایزومیر جدید شناسایی شد که در گونه‌هایی نظیر انسان و موش دارای عملکردهای مرتبط با ایمنی می‌باشند. بر اساس نتایج حاصل از این مطالعه ژن‌های هدف miRNAهای جدید در مسیرهای مرتبط با بیماری ورم پستان از جمله ایمنی، مرگ سلولی و التهاب نقش دارند و این مطلب می‌تواند مؤید نقش احتمالی و تنظیمی miRNA‌های جدید شناسایی شده در بروز ورم پستان باشد.

کلیدواژه‌ها

عنوان مقاله [English]

Identification and analysis of new miRNAs and isomirs and their targets in cows exposed to mastitis

چکیده [English]

Bovine mastitis is an inflammation disease of the mammary gland that impose considerable costs to the dairy industry. Regulatory mechanisms of this disease is complex and controlled by various gene regulatory elements and more studies are needed to better understand this disease. In the present study aimed to better understand of regulatory elements involved in mastitis, milk samples of two groups of healthy and infected cows during time series of 0, 12, 24, 36 and 48 hours after contamination were collected. The miRNA-seq data obtained from the milk samples and using the advanced bioinformatics, novel miRNAs, their targets and probability functions, isomirs and novel miRNAs* were identified. The results led to the identification of 92 novel miRNA including 26 miRNAs with homologous and 66 miRNAs without homologous genes in other species. Investigation of the functional groups of predicted targets genes, confirmed the roles of new miRNAs in response to internal and external stimulations, apoptosis and production of immunoglobulin. Furthermore, 135 novel miRNAs were identified. Also, 493 novel isomeric sibling miRNAs (isomers) were discovered that immune related functions of these isomirs were demonstrated in some species like human and mouse. Identification of miRNAs target genes with associated functions in mastitis, including safety, apoptosis and inflammation, can indicated the possible regulatory roles of the identified miRNAs in mastitis.

کلیدواژه‌ها [English]

  • Immune
  • inflammation
  • Mammary system
  • miRNA-seq
  • Next generation sequencing
  1.  

    1. Jin W, Ibeagha-Awemu EM, Liang G, Beaudoin F, Zhao X, et al. (2014) Transcriptome microRNA profiling of bovine mammary epithelial cells challenged with Escherichia coli or Staphylococcus aureus bacteria reveals pathogen directed microRNA expression profiles. BMC Genomics. 15: 181-183.
    2. Jin W, Ibeagha-Awemu EM, Liang G, Beaudoin F, Zhao X (2014) Transcriptome microRNA profiling of bovine mammary epithelial cells challenged with Escherichia coli or Staphylococcus aureus bacteria reveals pathogen directed microRNA expression profiles. BMC Genomics. 15: 1-3.
    3. Xu G, Gao Z, He W, Ma Y, Feng X, et al. (2014) microRNA expression in hepatitis B virus infected primary treeshrew hepatocytes and the independence of intracellular miR-122 level for de novo HBV infection in culture. Virology. 448: 247-254.
    4. Lu S, Sun Y-H and Chiang VL (2009) Adenylation of plant miRNAs. Nucleic Acids Research: gkp031.
    5. Naeem A, Zhong K, Moisá S, Drackley J, Moyes K, et al. (2012) Bioinformatics analysis of microRNA and putative target genes in bovine mammary tissue infected with Streptococcus uberis. Journal of Dairy Science. 95: 6397-6408.
    6. Gigli I and Maizon DO (2013) microRNAs and the mammary gland: a new understanding of gene expression. Genetics and Molecular Biology. 36: 465-474.
    7. Li R, Zhang C-L, Liao X-X, Chen D, Wang W-Q, et al. (2015) Transcriptome microRNA profiling of bovine mammary glands infected with Staphylococcus aureus. International Journal of Molecular Sciences. 16: 4997-5013.
    8. Lawless N, Reinhardt TA, Bryan K, Baker M, Pesch B, et al. (2014) MicroRNA regulation of bovine monocyte inflammatory and metabolic networks in an in vivo infection model. G3: Genes, Genomes, Genetics. 4: 957-971.
    9. Sturm M, Hackenberg M, Langenberger D and Frishman D (2010) TargetSpy: a supervised machine learning approach for microRNA target prediction. BMC Bioinformatics. 11: 292-309.
    10. Kertesz M, Iovino N, Unnerstall U, Gaul U and Segal E (2007) The role of site accessibility in microRNA target recognition. Nature Genetics. 39: 1278-1284.
    11. Le Guillou S, Marthey S, Laloë D, Laubier J, Mobuchon L, et al. (2014) Characterisation and comparison of lactating mouse and bovine mammary gland miRNomes. PloS One. 9: 3-8.
    12. Zheng Y, Chen K-l, Zheng X-m, Li H-x and Wang G-l (2014) Identification and bioinformatics analysis of microRNAs associated with stress and immune response in serum of heat-stressed and normal Holstein cows. Cell Stress and Chaperones. 19: 973-981.
    13. Hopster H, Van der Werf JT and Blokhuis HJ (1998) Stress enhanced reduction in peripheral blood lymphocyte numbers in dairy cows during endotoxin-induced mastitis. Veterinary Immunology and Immunopathology. 66: 83-97.
    14. Chang BS, Bohach GA, Lee S, Davis WC, Fox LK, et al. (2005) Immunosuppression by T regulatory cells in cows infected with Staphylococcal superantigen. Journal of Veterinary Science. 6: 247-249.
    15. Long E, Capuco A, Wood D, Sonstegard T, Tomita G, et al. (2001) Escherichia coli induces apoptosis and proliferation of mammary cells. Cell Death and Differentiation. 8: 808-816.
    16. Arias N, Aguirre L, Fernández-Quintela A, González M, Lasa A, et al. (2015) MicroRNAs involved in the browning process of adipocytes. Journal of Physiology and Biochemistry. 72: 509-521.
    17. Farrell D, Shaughnessy RG, Britton L, MacHugh DE, Markey B, et al. (2015) The Identification of Circulating MiRNA in Bovine Serum and Their Potential as Novel Biomarkers of Early Mycobacterium avium subsp paratuberculosis Infection. PloS One. 10: e0134310.
    18. Ma J, Li N, Guarnera M and Jiang F (2013) Quantification of plasma miRNAs by digital PCR for cancer diagnosis. Biomarker Insights. 8: 127-129.
    19. Qian B, Katsaros D, Lu L, Preti M, Durando A, et al. (2009) High miR-21 expression in breast cancer associated with poor disease-free survival in early stage disease and high TGF-β1. Breast Cancer Research and Treatment. 117: 131-140.
    20. Paraskevi A, Theodoropoulos G, Papaconstantinou I, Mantzaris G, Nikiteas N, et al. (2012) Circulating MicroRNA in inflammatory bowel disease. Journal of Crohn's and Colitis. 6: 900-904.
    21. Peng W-Z, Ma R, Wang F, Yu J and Liu Z-B (2014) Role of miR-191/425 cluster in tumorigenesis and diagnosis of gastric cancer. International Journal of Molecular Sciences. 15: 4031-4048.
    22. Watahiki A, Wang Y, Morris J, Dennis K, O'Dwyer HM, et al. (2011) MicroRNAs associated with metastatic prostate cancer. PloS One. 6: e24950.
    23. Johnnidis JB, Harris MH, Wheeler RT, Stehling-Sun S, Lam MH, et al. (2008) Regulation of progenitor cell proliferation and granulocyte function by microRNA-223. Nature. 451: 1125-1129.
    24. Yao K, He L, Gan Y, Zeng Q, Dai Y, et al. (2015) MiR-186 suppresses the growth and metastasis of bladder cancer by targeting NSBP1. Diagnostic Pathology. 10: 1-10.
    25. Würdinger T, Tannous BA, Saydam O, Skog J, Grau S, et al. (2008) miR-296 regulates growth factor receptor overexpression in angiogenic endothelial cells. Cancer Cell. 14: 382-393.