The generation of recombinant human WNT10A begins with the isolation and cloning of the gene that corresponds to the WNT10A protein (36-417aa). This target gene is inserted into a plasmid vector along with the
N-terminal 6xHis-tag gene, which is then introduced into yeast cells. These yeast cells are grown in bioreactors where the recombinant protein is produced. Following expression, the WNT10A protein is harvested from the cell lysates and purified using affinity chromatography. The SDS-PAGE is used to measure its purity, up to 90%.
WNT10A, a gene encoding a wingless pathway signaling molecule, is crucial in various developmental processes and cell cycle regulation [1]. It plays a significant role in tooth development, with defects in WNT10A being associated with tooth agenesis [2]. Studies have shown that WNT10A is essential for tooth root furcation morphogenesis and that mutations in this gene can lead to ectodermal dysplasia by affecting progenitor cell proliferation and differentiation [3][4]. Furthermore, WNT10A has been identified as a critical ligand controlling adult epithelial proliferation and differentiation, with downstream β-catenin pathway activation being a potential approach to address regenerative defects in WNT10A patients [4].
WNT10A has been linked to wound healing by regulating collagen expression and synthesis, as WNT10A-deficient mice exhibited delayed wound healing due to reduced collagen production [5]. The depletion of WNT10A has been shown to prevent tumor growth by suppressing microvessels and collagen expression [6]. WNT10A has been implicated in oncogenesis through the activation of the WNT/β-catenin signaling pathway in various cancers such as renal cell carcinoma, colorectal cancer, and ovarian cancer [7][8][9].
References:
[1] T. C, S. Jung, K. Niederreither, M. Prasad, S. Hadj-Rabia, N. Philipet al., Dental and extra‐oral clinical features in 41 patients with wnt10a gene mutations: a multicentric genotype–phenotype study, Clinical Genetics, vol. 92, no. 5, p. 477-486, 2017. https://doi.org/10.1111/cge.12972
[2] J. Yang, S. Wang, M. Choi, B. Reid, Y. Hu, Y. Leeet al., Taurodontism, variations in tooth number, and misshapened crowns inwnt10anull mice and human kindreds, Molecular Genetics & Genomic Medicine, vol. 3, no. 1, p. 40-58, 2014. https://doi.org/10.1002/mgg3.111
[3] M. Yu, Y. Liu, Y. Wang, S. Wong, J. Wu, H. Liuet al., Epithelialwnt10ais essential for tooth root furcation morphogenesis, Journal of Dental Research, vol. 99, no. 3, p. 311-319, 2020. https://doi.org/10.1177/0022034519897607
[4] M. Xu, J. Horrell, M. Snitow, J. Cui, H. Gochnauer, C. Syrettet al., Wnt10a mutation causes ectodermal dysplasia by impairing progenitor cell proliferation and klf4-mediated differentiation, Nature Communications, vol. 8, no. 1, 2017. https://doi.org/10.1038/ncomms15397
[5] K. Wang, S. Yamada, H. Izumi, M. Tsukamoto, T. Nakashima, T. Tasakiet al., Critical in vivo roles of wnt10a in wound healing by regulating collagen expression/synthesis in wnt10a-deficient mice, Plos One, vol. 13, no. 3, p. e0195156, 2018. https://doi.org/10.1371/journal.pone.0195156
[6] M. Kumagai, X. Guo, K. Wang, H. Izumi, M. Tsukamoto, T. Nakashimaet al., Depletion of wnt10a prevents tumor growth by suppressing microvessels and collagen expression, International Journal of Medical Sciences, vol. 16, no. 3, p. 416-423, 2019. https://doi.org/10.7150/ijms.26997
[7] J. Li, W. Zhang, L. Wang, & Y. Zhang, The oncogenic role of wnt10a in colorectal cancer through activation of canonical wnt/β‑catenin signaling, Oncology Letters, 2019. https://doi.org/10.3892/ol.2019.10035
[8] P. Li, W. Liu, Q. Xu, & C. Wang, Clinical significance and biological role of wnt10a in ovarian cancer, Oncology Letters, 2017. https://doi.org/10.3892/ol.2017.7062
[9] R. Hsu, J. Ho, T. Cha, D. Yü, C. Wu, W. Huanget al., Wnt10a plays an oncogenic role in renal cell carcinoma by activating wnt/β-catenin pathway, Plos One, vol. 7, no. 10, p. e47649, 2012. https://doi.org/10.1371/journal.pone.0047649
