Evaluation of Serum Amino Acid and Carnitine Profile in Dogs with Transmissible Venereal Tumor

Abstract

The presented study aimed to reveal the changes in serum amino acid and carnitine profiles in dogs with transmissible venereal tumor (TVT). The study material comprised 40 female dogs ranging in age from 3 to 5 years. The dogs were divided into two groups based on genital organ examinations. Group 1 (n=20) consisted of healthy dogs, while Group 2 (n=20) consisted of TVT-positive dogs. Blood samples were taken from dogs in both study groups, and serum was obtained. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for the determination of carnitine and amino acid profiles. The obtained data were compared using an independent samples t-test. The serum amino acid profiles of Lysine, Aspartic Acid, Tyrosine, Asparagine, Alanine, Arginine, Citrulline, Glutamic Acid, Glycine, Methylglutaryl, Phenylalanine, and Ornithine were found to be lower in the TVT group (P<0.05). The serum carnitine profiles of C0 (Free carnitine), C2 (Acetylcarnitine), C4 (Butyrylcarnitine), C4-DC (Methylmalonylcarnitine), C5 (Isovalerylcarnitine), C5:1 (Tiglylcarnitine), C5-OH (3-Hydroxyisovalerylcarnitine), C5-DC (Glutarylcarnitine), C6 (Hexanoylcarnitine), C6-DC (Adipoylcarnitine), C8 (Octanoylcarnitine), C8:1 (Octenoylcarnitine), C8-DC (Suberoylcarnitine), C10 (Decanoylcarnitine), C10:1 (Decenoylcarnitine), C12 (Dodecanoylcarnitine), C14 (Myristoylcarnitine), C14:1 (Myristoylcarnitine), C14:2 (Tetradecadienoylcarnitine), C18:1 (Oleoylcarnitine), C18:2 (Linoleylcarnitine), and C18:1-OH (Hydroxyoleylcarnitine) were found to be lower in the TVT group (P<0.01). The profiles of C3 (Propionylcarnitine) and C16 (Palmitoylcarnitine) were found to be higher in the TVT group (P<0.01). As a result, it was concluded that the significant changes in amino acid and carnitine values could be used as biomarkers for diagnosing TVT in dogs.

Keywords

• Amino Acid
• carnitine
• dog
• transmissible venereal tumor

Transmissible Venereal Tümörlü Köpeklerde Serum Amino Asit ve Karnitin Profilinin Değerlendirilmesi

Abstract

Sunulan çalışmada transmissible venereal tümörlü (TVT) köpeklerde serum amino asit ve karnitin profilindeki değişimleri ortaya koymak amaçlandı. Çalışma materyalini, 3-5 yaşları arasında değişen 40 adet dişi köpek oluşturdu. Köpekler genital organ muayenelerine göre iki gruba ayrıldı. Grup 1 (n=20) sağlıklı köpeklerden, Grup 2 (n=20) ise TVT pozitif köpeklerden oluştu. Her iki çalışma grubundaki köpeklerden kan örnekleri alınarak serum elde edildi. Karnitin ve amino asit profilinin belirlenmesinde, sıvı kromatografi-kütle spektrometresi (LC-MS/MS) kullanıldı. Elde edilen veriler bağımsız gruplar t-test ile karşılaştırıldı. Serum amino asitlerden Lizin, Aspartik Asit, Tirozin, Asparajin, Alanin, Arginin, Sitrülin, Glutamik Asit, Glisin, Metilglutaril, Fenilalanin ve Ornitin profilleri TVT grubunda daha düşük olduğu belirlendi (P<0.05). Serum karnitinlerden C0 (Serbest karnitin), C2 (Asetilkarnitin), C4 (Bütriylkarnitin), C4-DC (Metilmalonilkarnitin), C5 (İzovalerylkarnitin), C5:1 (Tiglikarnitin), C5-OH (3-Hidroksiizovalerylkarnitin), C5-DC (Glutarylkarnitin), C6 (Heksanoilkarnitin), C6-DC (Adipoilkarnitin), C8 (Oktanoilkarnitin), C8:1 (Oktenoylkarnitin), C8-DC (Suberoilkarnitin), C10 (Dekanoilkarnitin), C10:1 (Dekenoylkarnitin), C12 (Dodekanoilkarnitin), C14 (Miristoilkarnitin), C14:1 (Miristoilkarnitin), C14:2 (Tetradekadienoylkarnitin), C18:1 (Oleoylkarnitin), C18:2 (Linoleylkarnitin) ve C18:1-OH (Hidroksi-oleylkarnitin) profillerinin TVT grubunda daha düşük olduğu belirlendi (P<0.01). C3 (Propiyonilkarnitin) ve C16 (Palmitoilkarnitin) profilleri ise TVT grubunda daha yüksek tespit edildi (P<0.01). Sonuç olarak amino asit ve karnitin değerlerindeki belirgin değişimler bu iki profilin köpeklerde TVT teşhisinde biyobelirteç olarak kullanılabileceği kanısına varıldı.

Keywords

• Amino asit
• karnitin
• köpek
• transmissible venereal tümör

References

1. Abedin SN, 2020: Canine transmissible venereal tumor. J Entomol Zool Stud, 8(2), 596-599.
2. Aboud OA, Weiss RH, 2013: New opportunities from the cancer metabolome. Clin Chem, 59, 138-146.
3. Azuma K, Osaki T, Tsuka T, Imagawa T, Minami S, Okamoto Y, 2012: Plasma free amino acid profiles of canine mammary gland tumors. J Vet Sci, 13, 433.
4. Bogusiewicz J, Burlikowska K, Jaroch K, Gorynska PZ, Gorynski K, Birski M, Furtak J, Paczkowski D, Harat M, Bojko B, 2021: Profiling of carnitine shuttle system intermediates in gliomas using solid-phase microextraction (SPME). Molecules, 26(20), 6112.
5. Carracedo A, Cantley LC, Pandolfi PP, 2013: Cancer metabolism: Fatty acid oxidation in the limelight. Nat Rev Cancer, 13(4), 227-232.
6. Dambrova M, Makrecka-Kuka M, Kuka J, Vilskersts R, Nordberg D, Attwood M, Smesny S, Sen Z, Guo A, Oler E, Tian S, Zheng J, Wishart D, Liepinsh E, Schioth H, 2022: Acylcarnitines: nomenclature, biomarkers, therapeutic potential, drug targets, and clinical trials. Pharmacol Rev, 74, 506-551.
7. Das U, Das A, 2000: Review of canine transmissible venereal sarcoma. Vet Resear Comm, 24(8), 545-556. Dodson WL, Sachan DS, Krauss S, Hanna W, 1989: Alterations of serum and urinary carnitine profiles in cancer patients: hypothesis of possible significance. J Am Coll Nutr, 8, 133-142.
8. Green CR, Wallace M, Divakaruni AS, Phillips SA, Murphy AN, Ciaraldi TP, Metallo CM, 2016: Branched-chain amino acid catabolism fuels adipocyte differentiation and lipogenesis. Nat Chem Biol, 12, 15-21.

9. Hanahan D, Weinberg RA, 2011: Hallmarks of cancer: the next generation. Cell, 144, 646-74.
10. His M, Viallon V, Dossus L, Gicquiau A, Achaintre D, Scalbert A, Ferrari P, Romieu I, Moret NC, Weiderpass E, Dahm CC, Overvad K, Olsen A, Tjonneland A, Fournier A, Boeing H, Trichopoulou A, Nost TH, Karakatsani A, Martimianaki G, Masala G, Sieri S, Tumino R, Vineis P, Panico S, Gils HC, Sandanger TM, Skeie G, Quiros JR, Agudo A, Sanchez MJ, Amiano P, Huerta JM, Ardanaz E, Schmidt JA, Travis RC, Riboli E, Tsilidis KK, Christakoudi S, Gunter MJ, Rinaldi S, 2019: Prospective analysis of circulating metabolites and breast cancer in EPIC. BMC Med, 17(1), 178.
11. Houten SM, Wanders RJA, Ranea-Robles P, 2020: Metabolic interactions between peroxisomes and mitochondria with a special focus on acylcarnitine metabolism. Biochim Biophys Acta Mol Basis Dis, 1866(5), 165720.
12. Ihata Y, Miyagi E, Numazaki R, Muramatsu T, Imaizumi A, Yamamoto H, Yamakado M, Okamoto N, Hirahara F, 2014: Amino acid profile index for early detection of endometrial cancer: verification as a novel diagnostic marker. Int J Clin Oncol, 19, 364-372.
13. Jonker R, Engelen MPKJ, Deutz NEP, 2012: Role of specific dietary amino acids in clinical conditions. Br J Nutr, 108, 139-148.
14. Kawabe M, Baba Y, Tamai R, Yamamoto R, Komori M, Mori T, Takenaka S, 2015: Profiling of plasma metabolites in canine oral melanoma using gas chromatography-mass spectrometry. J Vet Med Sci, 77, 1025-1028.
15. Kim HH, Joo H, Kim TH, Kim EY, Park SJ, Park JK, Kim HJ, 2009: The mitochondrial Warburg effect: a cancer enigma. Interdisciplinary Bio Central, 1(2), 7.1-7.7.
16. Kozar N, Kruusmaa K, Bitenc M, Argamasilla R, Adsuar A, Takač I, Arko D, 2021: Identification of novel diagnostic biomarkers in breast cancer using targeted metabolomic profiling. Clin Breast Cancer, 21(3), 204-211.
17. Kubota A, Meguid MM, Hitch DC, 1992: Amino acid profiles correlate diagnostically with organ site in three kinds of malignant tumors. Cancer, 69(9), 2343-2348.
18. Lai HS, Lee JC, Lee PH, Wang ST, Chen WJ, 2005: Plasma free amino acid profile in cancer patients. Semin Cancer Biol, 15, 267-276.
19. Lieu EL, Nguyen T, Rhyne S, Kim J, 2020: Amino acids in cancer. Exp Mol Med, 52(1), 15-30.
20. Li D, Lu Y, Zhao F, Yan L, Yang X, Wei L, Yang X, Yuan X, Yang K, 2022: Targeted metabolomic profiles of serum amino acids and acylcarnitines related to gastric cancer. Peer J, 10, 14115.
21. Lu Y, Li N, Gao L, Xu YJ, Huang C, Yu K, Ling Q, Cheng Q, Chen S, Zhu M, Fang J, Chen M, Ong CN, 2016: Acetylcarnitine is a candidate diagnostic and prognostic biomarker of hepatocellular carcinoma. Cancer Res, 76, 2912-2920.
22. Mac-Ewen E, 2001: Transmissible venereal tumor. In: Withrow S and MacEwen E (eds.), Small Animal Clinical Oncology. 3rd Edition, Saunders, Philadelphia pp. 651-656.
23. Miyagi Y, Higashiyama M, Gochi A, Akaike M, Ishikawa T, Miura T, Saruki N, Bando E, Kimura H, Imamura F, Moriyama M, Ikeda I, Chiba A, Oshita F, Imaizumi A, Yamamoto H, Miyano H, Horimoto K, Tochikubo O, Mitsushima T, Yamakado M, Okamoto N, 2011: Plasma free amino acid profiling of five types of cancer patients and its application for early detection. PLoS One, 6, 24143.
24. McCann MR, George De la Rosa MV, Rosania GR, Stringer KA, 2021: L-carnitine and acylcarnitines: mitoLchondrial biomarkers for precision medicine. Metabolites, 11(1), 51.
25. Mukaratirwa S, Gruys E, 2003: Canine transmissible venereal tumour: cytogenetic origin, immunophenotype and immunobiology. Vet Quart, 25(3), 101-111.
26. Ni J, Xu L, Li W, Wu L, 2016: Simultaneous determination of thirteen kinds of amino acid and eight kinds of acylcarnitine in human serum by LC-MS/MS and its application to measure the serum concentration of lung cancer patients. Biomed Chromatogr, 30(11), 1796-1806.
27. Niziol J, Bonifay V, Ossolinski K, Ossolinski T, Ossolinska A, Sunner J, Beech I, Arendowski A, Ruman T, 2018: Metabolomic study of human tissue and urine in clear cell renal carcinoma by LC-HRMS and PLS-DA. Anal Bioanal Chem, 410, 3859-3869,
28. Pavlova NN, Thompson CB, 2016: The emerging hallmarks of cancer metabolism. Cell Metab, 23, 27-47.
29. Sachan DS, Datson WL, 1987: The serum carnitine status of cancer patients. J Am Coll Nutr, 6, 145-150.
30. Sandikci KS, Gumustas MK, Tuter Y, Kokoglu E, Ozyurt E, Sozer V, 1999: Total acid soluble and insoluble carnitine levels in human brain tumors. Cancer Biochem Biophys, 17, 49-57.
31. Schmidt M, Voelker HU, Kapp M, Krockenberger M, Dietl J, Kammerer U, 2010: Glycolytic phenotype in breast cancer: Activation of Akt, up-regulation of GLUT1, TKTL1 and down-regulation of M2PK. J Cancer Res Clin Oncol, 136, 219-225.
32. Shen Y, Sun M, Zhu J, Wei M, Li H, Zhao P, Wang J, Li R, Tian L, Tao Y, Shen P, Zhang J, 2021: Tissue metabolic profiling reveals major metabolic alteration in colorectal cancer, Mol Omics, 17, 464-471.
33. Tammo O, Uyanikoglu H, Koyuncu İ, 2021: Evaluation of plasma free amino acid and carnitine levels in patients with cesarean scar pregnancy. Comb Chem High Scr, 24(9), 1436-1445.
34. Tella MA, Ajala OO, Taiwo VO, 2004: Complete regression of transmissible venereal tumor (TVT) in Nigerian mongrel dogs with vincristine sulphate chemotherapy. Afr J Biomed Res, 7(3), 133-138.
35. Tochikubo O, Ando T, 2010: Amino acids and life-style related diseases: Application of new aminograms in life science, pp. 70-120, Kagawa Nutrition University Publishing Division, Tokyo.
36. Turkoglu O, Zeb A, Graham S, Szyperski T, Szender JB, Odunsi K, Bahado-Singh R, 2016: Metabolomics of biomarker discovery in ovarian cancer: a systematic review of the current literature. Metabolomics, 12, 60.
37. Vissers YLJ, Dejong CHC, Luiking YC, Fearon KCH, Meyenfeldt MF, Deutz NE, 2005: Plasma arginine concentrations are reduced in cancer patients: evidence for arginine deficiency? Am J Clin Nutr, 81, 1142-1146.
38. Ward PS, Thompson CB, 2012: Signaling in control of cell growth and metabolism. Cold Spring Harb Perspect Biol, 4(7), a006783.
39. Wolf M, Chen S, Zhao X, Scheler M, Irmler M, Staiger H, Beckers J, Angelis MH, Häring HU, Schleicher DE, Xu G, Lehmann R, Weigert C, 2013: Production and release of acylcarnitines by primary myotubes reflect the differences in fasting fat oxidation of the donors. J Clin Endocrinol Metab, 98(6), 1137-1142.
40. Xu J, Chen Y, Zhang R, Song Y, Cao J, Bi N, Wang J, He J, Bai J, Dong L, Wang L, Zhan, Z, Abliz Q, 2013: Global and targeted metabolomics of esophageal squamous cell carcinoma discovers potential diagnostic and therapeutic biomarkers. Mol Cell Proteomics, 12, 1306-1318.
41. Zhao F, An R, Wang L, Shan J, Wang X, 2021: Specific gut microbiome and serum metabolome changes in lung cancer patients. Front Cell Infect Microbiol, 11, 725284.