DOI QR코드

DOI QR Code

Bioinformatical Analysis of Messenger RNA and MicroRNA on Canine Splenic Tumors Based on Malignancy and Biopsy Sites

  • Eunpyo Kim (Department of Theriogenology and Biotechnology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University) ;
  • Giup Jang (Research and Development Center, Einocle Inc.) ;
  • Jin-Wook Kim (Research and Development Center, ROKIT Healthcare Inc.) ;
  • Wan-Hee Kim (Department of General Surgery, College of Veterinary Medicine, Seoul National University) ;
  • Geon-A Kim (Department of Biomedical Laboratory Science, School of Healthcare Science, Eulji University)
  • Received : 2023.01.02
  • Accepted : 2023.04.11
  • Published : 2023.04.30

Abstract

Canine splenic tumors (STs) are commonly diagnosed during imaging examinations, such as in X-ray and ultrasonography examinations, suggesting their higher prevalence, especially in older dogs. Despite this high prevalence, there are no effective treatment options for STs because of the difficulties in determining therapeutic targets. However, recently, the importance of microRNAs (miRNAs) has evolved owing to their ambivalent characteristics. Biomarkers and novel therapies using miRNAs have been well-studied in human cancer research compared to canine research, except for mammary gland tumors. Therefore, this study aimed to comparatively analyze miRNA expression profiles according to malignancy and biopsy sites to identify novel therapeutic and diagnostic targets. Tissue samples were collected directly from splenic tumor masses and immersed in RNAlater solution for further analysis. To investigate differentially expressed genes (DEGs) between tumor and normal tissues, we used RNA-seq and miRNA microarray analysis. Then, functional analysis based on DEGs was conducted to sort tumor-related DEGs. We found that cfa-miR-150 was upregulated in benign tumors, whereas cfa-miR-134 was upregulated in malignant tumors. Despite limited information on canine miRNAs, we identified two potential biomarkers for the differential diagnosis of STs.

Keywords

Acknowledgement

We specially thank the participated companion dogs and their owners for providing samples.

References

  1. Andrade AC, Freitas TR, Dornelas GG, Gomes LC, Barbosa BL, Araujo SS, et al. miR-197, miR-26a and miR-27a analysis in chronic lymphocytic leukemia. Biomark Med 2022; 16: 903-914. https://doi.org/10.2217/bmm-2021-0873
  2. Beuvink I, Kolb FA, Budach W, Garnier A, Lange J, Natt F, et al. A novel microarray approach reveals new tissue-specific signatures of known and predicted mammalian microRNAs. Nucleic Acids Res 2007; 35: e52.
  3. Carloni A, Terragni R, Morselli-Labate AM, Paninarova M, Graham J, Valenti P, et al. Prevalence, distribution, and clinical characteristics of hemangiosarcoma-associated skeletal muscle metastases in 61 dogs: a whole body computed tomographic study. J Vet Intern Med 2019; 33: 812-819. https://doi.org/10.1111/jvim.15456
  4. Chen Y, Wang X. miRDB: an online database for prediction of functional microRNA targets. Nucleic Acids Res 2020; 48(D1): D127-D131. https://doi.org/10.1093/nar/gkz757
  5. Davis AP, Grondin CJ, Johnson RJ, Sciaky D, Wiegers J, Wiegers TC, et al. Comparative Toxicogenomics Database (CTD): update 2021. Nucleic Acids Res 2021; 49(D1): D1138-D1143. https://doi.org/10.1093/nar/gkaa891
  6. Filipow S, Laczmanski L. Blood circulating miRNAs as cancer biomarkers for diagnosis and surgical treatment response. Front Genet 2019; 10: 169.
  7. Frixa T, Donzelli S, Blandino G. Oncogenic MicroRNAs: key players in malignant transformation. Cancers (Basel) 2015; 7: 2466-2485. https://doi.org/10.3390/cancers7040904
  8. Grimes JA, Prasad N, Levy S, Cattley R, Lindley S, Boothe HW, et al. A comparison of microRNA expression profiles from splenic hemangiosarcoma, splenic nodular hyperplasia, and normal spleens of dogs. BMC Vet Res 2016; 12: 272.
  9. Huang Z, Shi J, Gao Y, Cui C, Zhang S, Li J, et al. HMDD v3.0: a database for experimentally supported human microRNA-disease associations. Nucleic Acids Res 2019; 47(D1): D1013-D1017. https://doi.org/10.1093/nar/gky1010
  10. Kim D, Langmead B, Salzberg SL. HISAT: a fast spliced aligner with low memory requirements. Nat Methods 2015; 12: 357-360. https://doi.org/10.1038/nmeth.3317
  11. Lawrie CH, Gal S, Dunlop HM, Pushkaran B, Liggins AP, Pulford K, et al. Detection of elevated levels of tumour-associated microRNAs in serum of patients with diffuse large B-cell lymphoma. Br J Haematol 2008; 141: 672-675. https://doi.org/10.1111/j.1365-2141.2008.07077.x
  12. Molnar R, Szabo L, Tomesz A, Deutsch A, Darago R, Raposa BL, et al. The chemopreventive effects of polyphenols and coffee, based upon a DMBA mouse model with microRNA and mTOR gene expression biomarkers. Cells 2022; 11: 1300.
  13. O'Byrne K, Hosgood G. Splenic mass diagnosis in dogs undergoing splenectomy according to breed size. Vet Rec 2019; 184: 620.
  14. Pekarsky Y, Croce CM. Role of miR-15/16 in CLL. Cell Death Differ 2015; 22: 6-11. https://doi.org/10.1038/cdd.2014.87
  15. Peng Y, Croce CM. The role of MicroRNAs in human cancer. Signal Transduct Target Ther 2016; 1: 15004.
  16. Pertea M, Pertea GM, Antonescu CM, Chang TC, Mendell JT, Salzberg SL. StringTie enables improved reconstruction of a transcriptome from RNA-seq reads. Nat Biotechnol 2015; 33: 290-295. https://doi.org/10.1038/nbt.3122
  17. Pinero J, Queralt-Rosinach N, Bravo A, Deu-Pons J, Bauer-Mehren A, Baron M, et al. DisGeNET: a discovery platform for the dynamical exploration of human diseases and their genes. Database (Oxford) 2015; 2015: bav028.
  18. Pletscher-Frankild S, Palleja A, Tsafou K, Binder JX, Jensen LJ. DISEASES: text mining and data integration of disease-gene associations. Methods 2015; 74: 83-89. https://doi.org/10.1016/j.ymeth.2014.11.020
  19. Si W, Shen J, Zheng H, Fan W. The role and mechanisms of action of microRNAs in cancer drug resistance. Clin Epigenetics 2019; 11: 25.
  20. Thamm DH. Miscellaneous tumors. In: Withrow SJ, Vail DM, Page RL, editors. Withrow and MacEwen's small animal clinical oncology. 5th ed. St. Louis: Elsevier. 2013: 679-688.
  21. Weinstein MJ, Carpenter JL, Schunk CJ. Nonangiogenic and non-lymphomatous sarcomas of the canine spleen: 57 cases (1975-1987). J Am Vet Med Assoc 1989; 195: 784-788.
  22. Wendelburg KM, O'Toole TE, McCobb E, Price LL, Lyons JA, Berg J. Risk factors for perioperative death in dogs undergoing splenectomy for splenic masses: 539 cases (2001-2012). J Am Vet Med Assoc 2014; 245: 1382-1390. https://doi.org/10.2460/javma.245.12.1382
  23. Yan Y, Deng X, Ning X, Li F, Cao J. Pathogenic mechanism of miR21 in autoimmune lymphoid hyperplasia syndrome. Oncol Lett 2017; 13: 4734-4740. https://doi.org/10.3892/ol.2017.6039
  24. Yu G, Wang LG, Han Y, He QY. clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS 2012; 16: 284-287. https://doi.org/10.1089/omi.2011.0118
  25. Zhang B, Pan X, Cobb GP, Anderson TA. microRNAs as oncogenes and tumor suppressors. Dev Biol 2007; 302: 1-12. https://doi.org/10.1016/j.ydbio.2006.08.028