Bekas Nikolaos, Samiotaki Martina, Papathanasiou Maria, Mokos Panagiotis, Pseftogas Athanasios, Xanthopoulos Konstantinos, Thanos Dimitris, Mosialos George, Dafou Dimitra
School of Biology, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Biomedical Sciences Research Center "Alexander Fleming", 16672 Vari, Greece.
Cancers (Basel). 2023 Oct 15;15(20):4997. doi: 10.3390/cancers15204997.
is a tumor suppressor gene coding for a deubiquitinating enzyme that has a critical regulatory function in a variety of signaling pathways and biological processes involved in cancer development and progression, many of which are also key modulators of somatic cell reprogramming. Nevertheless, the potential role of in this process has not been studied. With the dual aim of investigating the involvement of in reprogramming and developing a better understanding of the intricate regulatory system governing this process, we reprogrammed control () and CYLD DUB-deficient () mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPSCs) through ectopic overexpression of the Yamanaka factors (Oct3/4, Sox2, Klf4, c-myc). CYLD DUB deficiency led to significantly reduced reprogramming efficiency and slower early reprogramming kinetics. The introduction of WT CYLD to MEFs rescued the phenotype. Nevertheless, CYLD DUB-deficient cells were capable of establishing induced pluripotent colonies with full spontaneous differentiation potential of the three germ layers. Whole proteome analysis (Data are available via ProteomeXchange with identifier PXD044220) revealed that the mesenchymal-to-epithelial transition (MET) during the early reprogramming stages was disrupted in MEFs. Interestingly, differentially enriched pathways revealed that the primary processes affected by CYLD DUB deficiency were associated with the organization of the extracellular matrix and several metabolic pathways. Our findings not only establish for the first time CYLD's significance as a regulatory component of early reprogramming but also highlight its role as an extracellular matrix regulator, which has profound implications in cancer research.
是一种肿瘤抑制基因，编码一种去泛素化酶，该酶在癌症发生和发展所涉及的多种信号通路和生物学过程中具有关键的调节功能，其中许多过程也是体细胞重编程的关键调节因子。然而，其在这一过程中的潜在作用尚未得到研究。为了研究其在重编程中的作用，并更好地理解控制这一过程的复杂调节系统，我们通过异位过表达山中因子（Oct3/4、Sox2、Klf4、c-myc），将对照（）和CYLD DUB缺陷（）小鼠胚胎成纤维细胞（MEFs）重编程为诱导多能干细胞（iPSCs）。CYLD DUB缺陷导致重编程效率显著降低，早期重编程动力学减慢。将野生型CYLD引入MEFs可挽救该表型。然而，CYLD DUB缺陷细胞能够建立具有三个胚层完全自发分化潜能的诱导多能集落。全蛋白质组分析（数据可通过ProteomeXchange获得，标识符为PXD044220）显示，早期重编程阶段的间充质向上皮转变（MET）在MEFs中被破坏。有趣的是，差异富集的通路显示，受CYLD DUB缺陷影响的主要过程与细胞外基质的组织和几种代谢通路有关。我们的研究结果不仅首次确立了CYLD作为早期重编程调节成分的重要性，还突出了其作为细胞外基质调节因子的作用，这对癌症研究具有深远意义。
