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Heparanase modulation by Wingless/INT (Wnt)

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Abstract

Heparanase is an endo-beta-glucuronidase, the only enzyme in mammals capable of cleaving heparan sulfate/heparin chains from proteoglycans. The oligosaccharides generated by heparanase present extensive biological functions since such oligosaccharides interact with adhesion molecules, growth factors, angiogenic factors and cytokines, modulating cell proliferation, migration, inflammation, and carcinogenesis. However, the regulation of heparanase activity is not fully understood. It is known that heparanase is synthesized as an inactive 65 kDa isoform and that post-translation processing forms an active 50 kDa enzyme. In the present study, we are interested in investigating whether heparanase is regulated by its own substrate as observed with many other enzymes. Wild-type Chinese hamster (Cricetulus griséus) ovary cells (CHO-K1) were treated with different doses of heparin. Heparanase expression was analyzed by Real-time PCR and flow cytometry. Also, heparanase activity was measured. The heparanase activity assay was performed using a coated plate with biotinylated heparan sulfate. In the present assay, a competitive heparin inhibition scenario was set aside. Exogenous heparin trigged a cell signaling pathway that increased heparanase mRNA and protein levels. The Wnt/beta-catenin pathway, judged by TCF-driven luciferase activity, seems to be involved to enhance heparanase profile during treatment with exogenous heparin. Lithium chloride treatment, an activator of the Wnt/beta-catenin pathway, confirmed such mechanism of transduction in vivo using zebrafish embryos and in vitro using CHO-K1 cells. Taken together the results suggest that heparin modulates heparanase expression by Wnt/beta-catenin.

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Acknowledgements

The authors would like to thank Dr. Jeffrey D. Esko, University of California San Diego and Dr. Mikael Peppelenbosch, Erasmus University Rotterdam, for kindly providing the CHO cell lines and pTOPFLASH and pFOPFLASH, respectively. In addition, the authors would like to FAPESP (São Paulo Research Foundation), CNPq (National Council for Technological and Scientific Development), and CAPES (Coordination for the Improvement of Higher Education Personnel) for financial support.

Funding

The financial support was obtained from FAPESP (São Paulo Research Foundation) grant, CNPq (National Council for Technological and Scientific Development), and CAPES (Coordination for the Improvement of Higher Education Personnel).

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Authors and Affiliations

  1. Department of Biochemistry, Universidade Federal de São Paulo, Rua Três de Maio, 100, 4a. andar, Biologia Molecular, São Paulo, SP, 04044-020, Brazil

    Carina Mucciolo Melo, Helena Bonciani Nader, Giselle Zenker Justo & Maria Aparecida Silva Pinhal

  2. Department of Biochemistry, Faculdade de Medicina do ABC, Avenida Príncipe de Gales, 821, Bioquímica, Santo André, SP, 09060-650, Brazil

    Carina Mucciolo Melo & Maria Aparecida Silva Pinhal

  3. Department of Biochemistry, Universidade Federal de São Paulo, Rua Prof. Artur Riedel, no. 275 - Jd. Eldorado, Diadema, SP, CEP: 09972-270, Brazil

    Giselle Zenker Justo

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  1. Carina Mucciolo Melo
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  3. Giselle Zenker Justo
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by CMM. The first draft of the manuscript was written by CMM and MASP. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Maria Aparecida Silva Pinhal.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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The authors hereby consent to publication of this study.

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All animals were treated according to the Universidade Federal de São paulo animal welfare guidelines as described and approved by UNIFESP Committee (2214150216).

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Melo, C.M., Nader, H.B., Justo, G.Z. et al. Heparanase modulation by Wingless/INT (Wnt). Mol Biol Rep 48, 3117–3125 (2021). https://doi.org/10.1007/s11033-021-06348-3

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  • DOI: https://doi.org/10.1007/s11033-021-06348-3

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