Department of Biological Chemistry, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
First Department of Pathology, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
J Cell Mol Med. 2022 Apr;26(8):2428-2437. doi: 10.1111/jcmm.17266. Epub 2022 Mar 13.
Craniosynostosis is the premature fusion of skull sutures and has a severe pathological impact on childrens' life. Mechanical forces are capable of triggering biological responses in bone cells and regulate osteoblastogenesis in cranial sutures, leading to premature closure. The mechanosensitive proteins polycystin-1 (PC1) and polycystin-2 (PC2) have been documented to play an important role in craniofacial proliferation and development. Herein, we investigated the contribution of PC1 to the pathogenesis of non-syndromic craniosynostosis and the associated molecular mechanisms. Protein expression of PC1 and PC2 was detected in bone fragments derived from craniosynostosis patients via immunohistochemistry. To explore the modulatory role of PC1 in primary cranial suture cells, we further abrogated the function of PC1 extracellular mechanosensing domain using a specific anti-PC1 IgPKD1 antibody. Effect of IgPKD1 treatment was evaluated with cell proliferation and migration assays. Activation of PI3K/AKT/mTOR pathway components was further detected via Western blot in primary cranial suture cells following IgPKD1 treatment. PC1 and PC2 are expressed in human tissues of craniosynostosis. PC1 functional inhibition resulted in elevated proliferation and migration of primary cranial suture cells. PC1 inhibition also induced activation of AKT, exhibiting elevated phospho (p)-AKT (Ser473) levels, but not 4EBP1 or p70S6K activation. Our findings indicate that PC1 may act as a mechanosensing molecule in cranial sutures by modulating osteoblastic cell proliferation and migration through the PC1/AKT/mTORC2 cascade with a potential impact on the development of non-syndromic craniosynostosis.
颅缝早闭是颅骨缝过早融合,严重影响儿童的生活。机械力能够在骨细胞中引发生物反应,并调节颅缝中的成骨细胞,导致过早闭合。多蛋白-1(PC1)和多蛋白-2(PC2)已被证明在颅面增殖和发育中发挥重要作用。在此,我们研究了 PC1 对非综合征性颅缝早闭发病机制的贡献及其相关分子机制。通过免疫组织化学检测来自颅缝早闭患者的骨片段中 PC1 和 PC2 的蛋白表达。为了探讨 PC1 在原发性颅缝细胞中的调节作用,我们进一步使用特异性抗 PC1 IgPKD1 抗体阻断 PC1 细胞外机械敏感结构域的功能。通过细胞增殖和迁移实验评估 IgPKD1 处理的效果。在 IgPKD1 处理后,通过 Western blot 进一步检测 PI3K/AKT/mTOR 通路成分的激活。PC1 和 PC2 在颅缝早闭患者的人组织中表达。PC1 功能抑制导致原发性颅缝细胞增殖和迁移增加。PC1 抑制还诱导 AKT 激活,表现为磷酸化 AKT(Ser473)水平升高,但不激活 4EBP1 或 p70S6K。我们的研究结果表明,PC1 可能通过 PC1/AKT/mTORC2 级联调节成骨细胞增殖和迁移,作为颅缝中的机械感知分子,对非综合征性颅缝早闭的发生发展有潜在影响。