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Characterization of Thrombospondin Type 1 Repeat in Haliotis diversicolor and Its Possible Role in Osteoinduction

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Abstract

Thrombospondin repeats (TSR) are important peptide domains present in the sequences of many extracellular and transmembrane proteins with which a variety of ligands interact. In this study, we characterized HdTSR domains in the ADAMTS3 protein of Thai abalone, Haliotis diversicolor, based on the transcriptomic analysis of its mantle tissues. PCR amplification and localization studies demonstrated the existence of HdTSR transcript and protein in H. diversicolor tissues, particularly in both the inner and outer mantle epithelial folds. We, therefore, generated a short recombinant protein, termed HdTSR1/2, based on the existence of the WxxWxxW or WxxxxW motif (which binds to TGF-β, a known signaling in bone formation/repair) in HdTSR1 and HdTSR2 sequences and used it to test the osteoinduction function in the pre-osteoblastic cell line, MC3T3-E1. This recombinant protein demonstrated the ability to induce the differentiation of MC3T3-E1 cells by the concentration- and time-dependent upregulation of many known osteogenic markers, including RUNX2, COL1A1, OCN, and OPN. We also demonstrated the upregulation of the SMAD2 gene after cell treatment with HdTSR1/2 proteinindicating its possible interaction through TGF-β, which thus activates its downstream signaling cascade and triggers the biomineralization process in the differentiated osteoblastic cells. Together, HdTSR domains existed in an extracellular ADAMTS3 protein in the mantle epithelium of H. diversicolor and played a role in osteoinduction as similar to the other nacreous proteins, opening up its possibility to be developed as an inducing agent of bone repair.

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References

  • Adams JC, Lawler J (2011) The thrombospondins. Cold Spring Harb Perspect Biol 3:a009712

  • Asch AS, Barnwell J, Silverstein RL, Nachman RL (1987) Isolation of the thrombospondin membrane receptor. J Clin Investig 79:1054–1061

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bekhouche M, Colig A (2015) The procollagen N-proteinases ADAMTS2, 3 and 14 in pathophysiology. Matrix Biol 44–46:46–53

    Article  PubMed  CAS  Google Scholar 

  • Bessa PC, Casal M, Reis RL (2008) Bone morphogenetic proteins in tissue engineering: the road from laboratory to clinic, part II (BMP delivery). J Tissue Eng Regen Med 2:81–96

    Article  CAS  PubMed  Google Scholar 

  • Buddawong T, Asuvapongpatana S, Senapin S, McDougall C, Weerachatyanukul W (2020) Characterization of calcineurin A and B genes in the abalone, Haliotis diversicolor, and their immune response role during bacterial infection. Peer J. https://doi.org/10.7717/peerj.8868

    Article  PubMed  PubMed Central  Google Scholar 

  • Chau JF, Leong WF, Li B (2009) Signaling pathways governing osteoblast proliferation, differentiation and function. Histol Histopathol 24:1593–1606

    CAS  PubMed  Google Scholar 

  • Chen G, Deng C, Li YP (2012) TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci 8:272–288

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen H, Herndon ME, Lawler J (2000) The cell biology of thrombospondin-1. Matrix Biol 19:597–614

    Article  CAS  PubMed  Google Scholar 

  • Crawford SE, Stellmach V, Murphy-Ullrich JE, Ribeiro SMF, Lawler J, Hynes RO, Boivin GP, Bouck N (1998) Thrombospondin-1 is a major activator of TGF-β1 in vivo. Cell 93:1159–1170

    Article  CAS  PubMed  Google Scholar 

  • Dawson DW, Pearce SF, Zhong R, Silverstein RL, Frazier WA, Bouck NP (1997) CD36 mediates the In vitro inhibitory effects of thrombospondin-1 on endothelial cells. J Cell Biol 138:707–717

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fernandes RJ, Hirohata S, Engle JM, Colige A, Cohn DH, Eyre DR, Apte SS (2001) Procollagen II amino propeptide processing by ADAMTS-3. Insights on Dermatosparaxis J Biol Chem 276:31502–31509

    Article  CAS  PubMed  Google Scholar 

  • Gardner L, Mills D, Wiegand A, Leavesley D, Elizur A (2011) Spatial analysis of biomineralization associated gene expression from the mantle organ of the pearl oyster Pinctana maxima. BMC Genomics 12:455

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gaume B, Fouchereau-peron M, Badou A, Helléouet MN, Huchette S, Auzoux-Bordenave S (2011) Biomineralization markers during early shell formation in the European abalone Haliotis tuberculata, Linnaeus. Mar Biol 158:341–353

    Article  CAS  Google Scholar 

  • Geoffroy V, Kneissel M, Fournier B, Boyde A, Matthias P (2002) High bone resorption in adult aging transgenic mice overexpressing cbfa1/runx2 in cells of the osteoblastic lineage. Mol Cell Biol 22:6222–6233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guo NH, Krutzsch HC, Negre E, Zabrenetzky VS, Roberts DD (1992) Heparin-binding peptides from the type I repeats of thrombospondin. Structural requirements for heparin binding and promotion of melanoma cell adhesion and chemotaxis. J Biol Chem 267:19349–19355

    Article  CAS  PubMed  Google Scholar 

  • Jang WG, Kim EJ, Kim DK, Ryoo HM, Lee KB, Kim SH, Choi HS, Koh JT (2012) J Biol Chem 287:905–915

    Article  CAS  PubMed  Google Scholar 

  • Janssens K, ten Dijke P, Janssens S, Hul VW (2005) Transforming growth factor-β1 to the bone. Endocr Rev 26:743–774

    Article  CAS  PubMed  Google Scholar 

  • Jardiller E, Rousseau M, Gendron-Badou A, Fröhlich F, Smith DC, Martin M, Helléouet MN, Huchette S, Doumenc D, Auzoux-Bordenave S (2008) A morphological and structural study of the larval shell from the abalone Haliotis tuberculata. Mar Biol 154:735–744

    Article  Google Scholar 

  • Jeltsch M, Jha SK, Tvorogov D, Anisimov A, Leppänen VM, Holopainen T, Kivelä R, Ortega S, Kärpanen T, Alitalo K (2014) CCBE1 enhances lymphangiogenesis via A disintegrin and metalloprotease with thrombospondin motifs-3-mediated vascular endothelial growth factor-C activation. Circulation 129:1962–1971

    Article  CAS  PubMed  Google Scholar 

  • Jolly C, Berland S, Milet C, Borzeix S, Lopez E, Doumenc D (2004) Zonal localization of shell matrix proteins in mantle of Haliotis tuberculata (Mollusca, Gastropoda). Mar Biotechnol 6:541–551

    Article  CAS  Google Scholar 

  • Kazimierczak P, Przekora A (2020) Osteoconductive and osteoinductive surface modifications of biomaterials for bone regeneration: A Concise Review. Coatings 10–00971:1–25

    Google Scholar 

  • Khoshniat S, Bourgine A, Julien M, Petit M, Pilet P, Rouillon T, Masson M, Gatius M, Weiss P, Guicheux J, Beck L (2011) Phosphate-dependent stimulation of MGP and OPN expression in osteoblasts via the ERK1/2 pathway is modulated by calcium. Bone 48:894–902

    Article  CAS  PubMed  Google Scholar 

  • Kirkham G, Cartmell S (2007) In: Ashammakhi N, Reis R, Chiellini E (eds) Topics in tissue engineering Vol 3. Oulu University, Finland

  • Klar A, Baldassare M, Jessell TM (1992) F-spondin: a gene expressed at high levels in the floor plate encodes a secreted protein that promotes neural cell adhesion and neurite extension. Cell 69:95–110

    Article  CAS  PubMed  Google Scholar 

  • Kilpelainen I, Kaksonen M, Kinnunen T, Avikainen H, Fath M, Linhardt RJ, Raulo E, Rauvala H (2000) Heparin-binding growth-associated molecule contains two heparin-binding beta-sheet domains that are homologous to the thrombospondin type I repeat. J Biol Chem 275:13564–13570

    Article  CAS  PubMed  Google Scholar 

  • Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T (1997) Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 89:755–764

    Article  CAS  PubMed  Google Scholar 

  • Lawler J, Connolly JE, Ferro P, Derick LH (1986) Thrombin and chymotrypsin interactions with thrombospondina. Ann N Y Acad Sci 485:273–287

    Article  CAS  PubMed  Google Scholar 

  • Le Goff C, Somerville RP, Kesteloot F, Powell K, Birk DE, Colige AC, Apte SS (2006) Regulation of procollagen amino-propeptide processing during mouse embryogenesis by specialization of homologous ADAMTS proteases: insights on collagen biosynthesis and dermatosparaxis. Development 133:1587–1596

    Article  PubMed  CAS  Google Scholar 

  • Lee KS, Hong SH, Bae SC (2002) Both the Smad and p38 MAPK pathways play a crucial role in Runx2 expression following induction by transforming growth factor-beta and bone morphogenetic protein. Oncogene 21:7156–7163

    Article  CAS  PubMed  Google Scholar 

  • Liedert A, Kaspar D, Blakytny R, Claes L, Ignatius A (2006) Signal transduction pathways involved in mechanotransduction in bone cells. Biochem Biophys Res Commun 349:1–5

    Article  CAS  PubMed  Google Scholar 

  • Liu C, Li S, Huang J, Liu Y, Jia G, Xie L, Zhang R (2015a) Extensible byssus of Pinctada fucata: Ca2+-stabilized nanocavities and a thrombospondin-1 protein. Sci Rep 5:15018

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu C, Li S, Kong J, Liu Y, Wang T, Xie L, Zhang R (2015b) In-depth proteomic analysis of shell matrix proteins of Pinctada fucata. Sci Rep 5:17269

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lopez-Dee Z, Pidcock K, Gutierrez LS (2011) Thrombospondin-1: multiple paths to inflammation. Mediators Inflamm. https://doi.org/10.1155/2011/296069

    Article  PubMed  PubMed Central  Google Scholar 

  • Ma JY, Wong KL, Xu ZY, Au KY, Lee NL, Su C, Su WW, Li PB, Shaw PC (2016) N16, a nacreous protein, inhibits osteoclast differentiation and enhances osteogenesis. J Nat Prod 79:204–212

    Article  CAS  PubMed  Google Scholar 

  • Massague J, Seoane J, Wotton D (2005) Smad transcription factors. Genes Dev 19:2783–2810

    Article  CAS  PubMed  Google Scholar 

  • Miyamoto H, Miyoshi F, Kohno J (2005) The carbonic anhydrase domain protein nacrein is expressed in the epithelial cells of the mantle and acts as a negative regulator in calcification in the mollusc Pinctada fucata. Zool Sci 22:311–315

    Article  CAS  Google Scholar 

  • Mori K, Kitazawa R, Kondo T, Maeda S, Yamaguchi A, Kitazawa S (2006) Modulation of mouse RANKL gene expression by Runx2 and PKA pathway. J Cell Biochem 98:1629–1644

    Article  CAS  PubMed  Google Scholar 

  • Murphy-Ullrich JE, Poczatek M (2000) Activation of latent TGF-beta by thrombospondin-1: mechanisms and physiology. Cytokine Growth Factor Rev 11:59–69

    Article  CAS  PubMed  Google Scholar 

  • Murphy-Ullrich JE, Schultz-Cherry S, Höök M (1992) Transforming growth factor-beta complexes with thrombospondin. Mol Biol Cell 3:181–188

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR (2002) The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 108:17–29

    Article  CAS  PubMed  Google Scholar 

  • Sato C, Fukuoka H, Ohta K, Matsuda T, Koshino R, Kobayashi K, Troy FA, Kitajima K (2000) Frequent occurrence of pre-existing alpha 2–>8-linked disialic and oligosialic acids with chain lengths up to 7 Sia residues in mammalian brain glycoproteins. Prevalence revealed by highly sensitive chemical methods and anti-di-, oligo-, and poly-Sia antibodies specific for defined chain lengths. J Biol Chem 275:15422–15431

    Article  CAS  PubMed  Google Scholar 

  • Schultz-Cherry S, Chen H, Mosher DF, Misenheimer TM, Krutzsch HC, Roberts DD, Murphy-Ullrich JE (1995) Regulation of transforming growth factor-beta activation by discrete sequences of thrombospondin 1. J Biol Chem 270:7304–7310

    Article  CAS  PubMed  Google Scholar 

  • Selvamurugan N, Kwok S, Alliston T, Reiss M, Partridge NC (2004) Transforming growth factor-β1 regulation of collagenase-3 expression in osteoblastic cells by cross-talk between the Smad and MAPK signaling pathways and their components, Smad2 and Runx2. J Biol Chem 279:19327–19334

    Article  CAS  PubMed  Google Scholar 

  • Shin MK, Kim M, Bae YS, Jo I, Lee SJ, Chung CP, Park YJ, Min DS (2008) A novel collagen-binding peptide promotes osteogenic differentiation via Ca2+/calmodulin-dependent protein kinase II/ERK/AP-1 signaling pathway in human bone marrow-derived mesenchymal stem cells. Cell Signal 20:613–624

    Article  CAS  PubMed  Google Scholar 

  • Silverstein RL, Baird M, Lo SK, Yesner LM (1992) Sense and antisense cDNA transfection of CD36 (glycoprotein IV) in melanoma cells. Role of CD36 as a thrombospondin receptor. J Biol Chem 267:16607–16612

    Article  CAS  PubMed  Google Scholar 

  • Silverstein RL, Asch AS, Nachman RL (1989) Glycoprotein IV mediates thrombospondin-dependent platelet-monocyte and platelet-U937 cell adhesion. J Clin Invest 84:546–552

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song X, Wang X, Li L, Zhang G (2014) Identification two novel nacrein-like proteins involved in the shell formation of the Pacific oyster Crassostrea gigas. Mol Biol Rep 41:4273–4278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Takeuchi T, Yamada L, Shinzato C, Sawada H, Satoh N (2016) Stepwise evolution of coral biomineralization revealed with genome-wide proteomics and transcriptomics. PLoS One 11: e0156424

  • Takeuchi T, Endo K (2006) Biphasic and dually coordinated expression of the genes encoding major shell matrix proteins in the pearl oyster Pinctada fucata. Mar Biotechnol 8:52–61

    Article  CAS  Google Scholar 

  • Tan K, Duquette M, Liu JH, Dong Y, Zhang R, Joachimiak A, Lawler J, Wang JH (2002) Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. J Cell Biol 159:373–382

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tucker RP (2004) The thrombospondin type 1 repeat superfamily. Int J Biochem Cell Biol 36:969–974

    Article  CAS  PubMed  Google Scholar 

  • Werner GD, Gemmell P, Grosser S, Hamer R, Shimeld SM (2013) Analysis of a deep transcriptome from the mantle tissue of Patella vulgata Linnaeus (Mollusca: Gastropoda: Patellidae) reveals candidate biomineralising genes. Mar Biotechnol 15:230–243

    Article  CAS  Google Scholar 

  • Wu M, Chen G, Li YP (2016) TGF-β and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease. Bone Research 4:16009

    Article  PubMed  PubMed Central  Google Scholar 

  • Xu ZY, Liu YL, Lin JB, Cheng KL, Wang YG, Yao HL, Peng W, Wu HY, Su WW, Shaw PC, Li PB (2018) Preparative expression and purification of a nacreous protein N16 and testing its effect on osteoporosis rat model. Int J Biol Macromol 111:440–445

    Article  CAS  PubMed  Google Scholar 

  • Young GD, Murphy-Ullrich JE (2004) The tryptophan-rich motifs of the thrombospondin type 1 repeats bind VLAL motifs in the latent transforming growth factor-beta complex. J Biol Chem 279:47633–47642

    Article  CAS  PubMed  Google Scholar 

  • Zainabadi K, Liu CJ, Guarente L (2017) SIRT1 is a positive regulator of the master osteoblast transcription factor, RUNX2. PLoS One. 12:e0178520

  • Zhao L, Jiang S, Hantash BM (2010) Transforming growth factor beta1 induces osteogenic differentiation of murine bone marrow stromal cells. Tissue Eng Part A 16:725–733

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank the Center of Nanoimaging (CNI) and the Central Instrument Facility (CIF), Faculty of Science, Mahidol University, for providing instrumental support throughout this work.

Funding

This research project is supported by Mahidol University (to SA).

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Author notes

  1. Valainipha Habuddha and Chanyatip Suwannasing are co-first authors of this paper.

Authors and Affiliations

  1. Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand

    Valainipha Habuddha, Aticha Buddawong, Chanyarak Sombutkayasith, Wattana Weerachatyanukul & Somluk Asuvapongpatana

  2. School of Allied Health Science, Walailak University, Nakhon Si Thammarat, Thailand

    Valainipha Habuddha

  3. Department of Radiological Technology, Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, Thailand

    Chanyatip Suwannasing

  4. Chulabhorn International College of Medicine, Thammasat University, Khlong Nueng Pathumthani 12121, Rangsit Campus, Thailand

    Aticha Buddawong

  5. Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University (Salaya Campus), Salaya, Nakhonpathom, Thailand

    Kanokwan Seenprachawong, Thitinat Duangchan & Aungkura Supokawej

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  1. Valainipha Habuddha
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Correspondence to Somluk Asuvapongpatana.

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The study was carried out in accordance with the protocol approved by the Animal Care Committee, Faculty of Science, Mahidol University (Protocol # MUSC-60–040-390).

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Habuddha, V., Suwannasing, ., Buddawong, A. et al. Characterization of Thrombospondin Type 1 Repeat in Haliotis diversicolor and Its Possible Role in Osteoinduction. Mar Biotechnol 23, 641–652 (2021). https://doi.org/10.1007/s10126-021-10054-3

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