CYLDis 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 ofCYLDin this process has not been studied. With the dual aim of investigating the involvement ofCYLDin reprogramming and developing a better understanding of the intricate regulatory system governing this process, we reprogrammed control (CYLDWT/WT) and CYLD DUB-deficient (CYLDΔ9/Δ9) 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 toCYLDΔ9/Δ9MEFs 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 inCYLDΔ9/Δ9MEFs. 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.
CYLD是一种肿瘤抑制基因，其编码的去泛素化酶在癌症发生发展相关的多种信号通路和生物过程中具有关键调控功能，其中许多通路同时也是体细胞重编程的关键调节因子。然而，CYLD在此过程中的潜在作用尚未得到研究。为探究CYLD在重编程中的参与机制，并深入理解这一过程的复杂调控系统，我们通过异位过表达山中因子（Oct3/4、Sox2、Klf4、c-myc），将对照组（CYLDWT/WT）与CYLD去泛素化功能缺失型（CYLDΔ9/Δ9）小鼠胚胎成纤维细胞重编程为诱导多能干细胞。CYLD去泛素化功能缺失导致重编程效率显著降低，早期重编程动力学进程减缓。通过向CYLDΔ9/Δ9细胞回补野生型CYLD可逆转该表型。值得注意的是，CYLD去泛素化功能缺失细胞仍能形成具备三胚层完全自发分化潜能的诱导多能干细胞集落。全蛋白质组分析（数据可通过ProteomeXchange获取，标识符PXD044220）显示，CYLDΔ9/Δ9细胞在早期重编程阶段的间充质-上皮转化过程发生紊乱。有趣的是，差异富集通路分析表明，受CYLD去泛素化功能缺失影响的主要过程与细胞外基质重构及多条代谢通路相关。本研究不仅首次确立了CYLD作为早期重编程调控元件的重要性，同时揭示了其作为细胞外基质调节因子的新功能，这对癌症研究具有深远意义。
Inactivation of Tumor Suppressor CYLD Inhibits Fibroblast Reprogramming to Pluripotency
肿瘤(癌症)患者之家