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The involvement of RUNX2 and SPARC genes in the bacterial chondronecrosis with osteomyelitis in broilers

  • E. Paludo (a1), A. M. G. Ibelli (a2), J. O. Peixoto (a2), F. C. Tavernari (a2), C. A. V. Lima-Rosa (a1), J. R. C. Pandolfi (a2) and M. C. Ledur (a2)...


Economic losses due to an increase of leg disorders in broilers have become a major concern of the poultry industry. Despite the efforts to reduce skeletal abnormalities in chickens, insufficient progress has been made. Bacterial chondronecrosis with osteomyelitis (BCO) is one of the main disorders that affect bone integrity in broilers. However, the genetic pathways and genes involved in most bone problems, including BCO, remains unclear. In this study, femoral samples from male broilers with 45 days of age affected or not with BCO were used to compare the relative expression with a reverse transcription real time PCR approach of 13 candidate genes: SPP1 (osteopontin), TNFRSF11B (osteoprotegerin), SPARC (osteonectin), CALB1 (calbidin 1), CALM (Calmodulin 2), IBSP (sialoprotein), COL1A2 (collagen, type I, α 2), BMP2 (bone morphogenetic protein 2), BMP3 (bone morphogenetic protein 3), RANKL (κ-B nuclear factor ligand), SMAD1 (SMAD family member 1), LEPR (leptin receptor) and RUNX2 (related transcription factor Runt 2). Differential expression test between affected and non-affected groups was performed using the REST software. The RUNX2 and SPARC genes were downregulated (P<0.05) in the affected group, with reduced expression of fourfold when compared with the non-affected group. This result indicates that the downregulation of RUNX2 and SPARC can contribute to an increased incidence of BCO in broilers.


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Bandow, K, Maeda, A, Kakimoto, K, Kusuyama, J, Shamoto, M and Ohnishi, T 2010. Molecular mechanisms of the inhibitory effect of lipopolysaccharide (LPS) on osteoblast differentiation. Biochemical and Biophysical Research Communications 402, 755761.
Bishop, SC, Fleming, RH, Mccormack, HA, Flock, DK and Whitehead, CC 2000. Inheritance of bone characteristics affecting osteoporosis in laying hens. British Poultry Science 1668, 3340.
Boskey, AL, Moore, DJ, Amling, M, Canalis, E and Delany, AM 2003. Infrared analysis of the mineral and matrix in bones of osteonectin-null mice and their wildtype controls. Journal of Bone and Mineral Research 18, 10051011.
Carbonare, LD, Matte, A, Valenti, MT, Siciliano, A, Mori, A, Schweiger, V, Zampieri, G, Perbellini, L and De Franceschi, L 2015. Hypoxia-reperfusion affects osteogenic lineage and promotes sickle cell bone disease. Blood 126, 23202328.
Cook, ME 2000. Skeletal deformities and their causes: introduction. Poultry Science 79, 982984.
Cydzik, M, Abdul-Wahid, A, Park, S, Bourdeau, A, Bowden, K, Prodeus, A, Kollara, A, Brown, TJ, Ringuette, MJ and Gariepy, J 2015. Slow binding kinetics of secreted protein, acidic, rich in cysteine-VEGF interaction limit VEGF activation of VEGF receptor 2 and attenuate angiogenesis. The FASEB Journal 29, 34933505.
Delany, AM, Amling, M, Priemel, M, Howe, C, Baron, R and Canalis, E 2000. Osteopenia and decreased bone formation in osteonectin-deficient mice. Journal of Clinical Investigation 105, 915923.
Fornari, MB, Zanella, R, Ibelli, AM, Fernandes, LT, Cantão, ME, Thomaz-Soccol, V, Ledur, MC and Peixoto, JO 2014. Unraveling the associations of osteoprotegerin gene with production traits in a paternal broiler line. SpringerPlus 3, 682.
Fujita, T, Azuma, Y, Fukuyama, R, Hattori, Y, Yoshida, C, Koida, M, Ogita, K and Komori, T 2004. Runx2 induces osteoblast and chondrocyte differentiation and enhances their migration by coupling with PI3K-Akt signaling. The Journal of Cell Biology 166, 8595.
Havenstein, GB, Ferket, PR, Scheideler, SE and Larson, BT 2003. Growth, livability, and feed conversion of 1957 vs 1991 broilers when fed ‘typical’ 1957 and 1991 broiler diets. Poultry Science 73, 17851794.
Jiang, T, Mandal, RK, Wideman, RF, Khatiwara, A, Pevzner, I and Min Kwon, Y 2015. Molecular survey of bacterial communities associated with bacterial chondronecrosis with osteomyelitis (BCO) in broilers. PLoS One 10, e0124403.
Johnsson, M, Jonsson, KB, Andersson, L, Jensen, P and Wright, D 2015. Genetic regulation of bone metabolism in the chicken: similarities and differences to mammalian systems. PLoS Genetics 11, e1005250.
Karsenty, G 2001. Minireview: transcriptional control of osteoblast differentiation. Endocrinology 142, 27312733.
Kato, Y, Lewalle, JM, Baba, Y, Tsukuda, M, Sakai, N, Baba, M, Kobayashi, K, Koshika, S, Nagashima, Y, Frankenne, F, Noël, A, Foidart, JM and Hata, RI 2001. Induction of SPARC by VEGF in human vascular endothelial cells. Biochemical and Biophysical Research Communications 287, 422426.
Kido, HW, Bossini, PS, Tim, CR, Parizotto, NA, da Cunha, AF, Malavazi, I and Renno, ACM 2014. Evaluation of the bone healing process in an experimental tibial bone defect model in ovariectomized rats. Aging Clinical and Experimental Research 26, 473481.
Kwon, TG, Zhao, X, Yang, Q, Li, Y, Ge, C, Zhao, G and Franceschi, RT 2011. Physical and functional interactions between Runx2 and HIF-1α induce vascular endothelial growth factor gene expression. Journal of Cellular Biochemistry 112, 35823593.
Li, PF, Zhou, ZL, Shi, CY and Hou, JF 2015. Downregulation of basic fibroblast growth factor is associated with femoral head necrosis in broilers. Poultry Science 94, 10521059.
Ma, X, Liu, Z, Ma, J, Han, C and Zang, J 2010. Dynamic expression of Runx2, Osterix and AJ18 in the femoral head of steroid-induced osteonecrosis in rats. Orthopaedic Surgery 2, 278284.
Mackie, EJ, Ahmed, YA, Tatarczuch, L, Chen, KS and Mirams, M 2008. Endochondral ossification: how cartilage is converted into bone in the developing skeleton. The International Journal of Biochemistry & Cell Biology 40, 4662.
Mandal, RK, Jiang, T, Al-Rubaye, AA, Rhoads, DD, Wideman, RF, Zhao, J, Pevzner, I and Kwon, YM 2016. An investigation into blood microbiota and its potential association with bacterial chondronecrosis with osteomyelitis (BCO) in broilers. Scientific Reports 6, 25882.
McNamee, PT and Smyth, JA 2000. Bacterial chondronecrosis with osteomyelitis (‘femoral head necrosis’) of broiler chickens: a review. Avian Pathology 29, 477495.
Mendoza-Londono, R, Fahiminiya, S, Majewski, J, Tétreault, M, Nadaf, J, Kannu, P, Sochett, E, Howard, A, Stimec, J, Dupuis, L, Roschger, P, Klaushofer, K, Palomo, T, Ouellet, J, Al-Jallad, H, Mort, JS, Moffatt, P, Boudko, S, Bächinger, HP and Rauch, F 2015. Recessive osteogenesis imperfecta caused by missense mutations in SPARC. The American Journal of Human Genetics 96, 979985.
Mori, G, Amelio, PD, Faccio, R and Brunetti, G 2013. The interplay between the bone and the immune system. Clinical and Developmental Immunology 2013, 720504.
Nozaki, M, Sakurai, E, Raisler, BJ, Baffi, JZ, Witta, J, Ogura, Y, Brekken, RA, Sage, EH and Ambati, J 2006. Loss of SPARC-mediated VEGFR-1 suppression after injury reveals a novel antiangiogenic activity of VEGF-A. Journal of Clinical Investigation 116, 422429.
Orlando, B, Giacomelli, L, Ricci, M, Barone, A and Covani, U 2013. Leader genes in osteogenesis: a theoretical study. Archives of Oral Biology 58, 4249.
Park, O, Kim, J, Yang, J and Yun, C 2015. Enterococcus faecalis inhibits osteoblast differentiation and induces chemokine expression. Journal of Endodontics 41, 49.
Parvaneh, K, Ebrahimi, M, Sabran, MR, Karimi, G, Hwei, ANM, Abdul-Majeed, S, Ahmad, Z, Ibrahim, Z and Jamaluddin, R 2015. Probiotics (Bifidobacterium longum) increase bone mass density and upregulate SPARC and BMP-2 genes in rats with bone loss resulting from ovariectomy. BioMed Research International 2015, 110.
Pfaffl, MW, Horgan, GW and Dempfle, L 2002. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Research 30, e36.
Poniatowski, ŁA, Wojdasiewicz, P, Gasik, R and Szukiewicz, D 2015. Transforming growth factor Beta family: insight into the role of growth factors in regulation of fracture healing biology and potential clinical applications. Mediators of Inflammation 2015, 137823.
Pugin, J 2012. How tissue injury alarms the immune system and causes a systemic inflammatory response syndrome. Annals of Intensive Care 2, 27.
Rath, NC, Huff, GR, Huff, WE and Balog, JM 2000. Factors regulating bone maturity and strength in poultry. Poultry Science 79, 10241032.
Reed, MJ, Bradshaw, AD, Shaw, M, Sadoun, E, Han, N, Ferara, N, Funk, S, Puolakkainen, P and Sage, EH 2005. Enhanced angiogenesis characteristic of SPARC-null mice disappears with age. Journal of Cellular Physiology 204, 800807.
Ribeiro, N, Sousa, SR, Brekken, RA. and Monteiro, FJ 2014. Role of SPARC in bone remodeling and cancer-related bone metastasis. Journal of Cellular Biochemistry 115, 1726.
Ruijter, JM, Ramakers, C, Hoogaars, WMH, Karlen, Y, Bakker, O, van den Hoff, MJB and Moorman, AFM 2009. Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Research 37, e45.
Sanchez, CJ Jr, Ward, CL, Romano, DR, Hurtgen, BJ, Hardy, SK, Woodbury, RL, Trevino, AV, Rathbone, CH and Wenke, JC 2013. Staphylococcus aureus biofilms decrease osteoblast viability, inhibits osteogenic differentiation and increases bone resorption in vitro. BMC Musculoskeletal Disorders 14, 187.
Szklarczyk, D, Franceschini, A, Wyder, S, Forslund, K, Heller, D, Huerta-Cepas, J, Simonovic, M, Roth, A, Santos, A, Tsafou, KP, Kuhn, M, Bork, P, Jensen, LJ and Von Mering, C 2015. STRING v10: protein-protein interaction networks, integrated over the tree of life. Nucleic Acids Research 43, D447D452.
Wideman, RF, Hamal, KR, Stark, JM, Blankenship, J, Lester, H, Mitchell, KN, Lorenzoni, G and Pevzner, I 2012. A wire-flooring model for inducing lameness in broilers: evaluation of probiotics as a prophylactic treatment. Poultry Science 91, 870883.
Wideman, RF and Prisby, RD 2013. Bone circulatory disturbances in the development of spontaneous bacterial chondronecrosis with osteomyelitis: a translational model for the pathogenesis of femoral head necrosis. Frontiers in Endocrinology 3, 114.
Wu, J, Pines, M, Gay, CV, Hurwitz, S and Leach, RM Jr 1996. Immunolocalization of osteonectin in avian tibial dyschondroplastic cartilage. Developmental Dynamics 207, 6974.
Xie, F, Xiao, P, Chen, D, Xu, L and Zhang, B 2012. miRDeepFinder: a miRNA analysis tool for deep sequencing of plant small RNAs. Plant Molecular Biology 80, 7584.
Ye, J, Coulouris, G, Zaretskaya, I, Cutcutache, I, Rozen, S and Madden, TL 2012. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13, 134.
Zelzer, E, Glotzer, DJ, Hartmann, C, Thomas, D, Fukai, N, Soker, S and Olsen, BR 2001. Tissue specific regulation of VEGF expression during bone development requires Cbfa1/Runx2. Mechanisms of Development 106, 97106.
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