Laragione Teresina, Harris Carolyn, Gulko Percio S
Division of Rheumatology, Department of Medicine, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, Box 1244, New York, NY, 10029, USA.
Sci Rep. 2025 Jul 11;15(1):25164. doi: 10.1038/s41598-025-08960-w.
The fibroblast-like synoviocyte (FLS) has a central role in rheumatoid arthritis (RA) pathogenesis and its invasive behavior strongly correlates with disease severity and joint damage. Yet, the regulation of FLS invasiveness is incompletely understood. Distal-less homeobox 4 (DLX4) is a transcription factor implicated in cancer cell invasion and metastasis, and we considered that it might also be involved in the regulation of FLS phenotypes. siRNA was used to knockdown DLX4 in RA FLS, compared with a control siRNA. Cells were then studied in invasion, migration, proliferation and adhesion assays, and RNA was used for RNA sequencing and pathway analyses. siRNA knockdown of DLX4 significantly reduced RA FLS invasiveness (P = 0.028) and migration in the scratch/wound healing assay (P = 0.008). RNA sequencing analyses revealed that DLX4 knockdown significantly affected processes and pathways implicated genes involved in diverse types of cancer and in cancer biology, including cell invasion, cell cycle, DNA replication, transcription, p53 signal transduction, RHO GTPase signaling, regulation of cytokinesis, as well as response to oxygen levels and to oxidative stress. HMOX1 and SOD2 were among the genes with the most significantly increased expression, while CPNE3 and HADHA were among those with the most significantly decreased expression in FLS knocked down for DLX4 compared with controls. We describe a new role for DLX4 in the regulation of RA FLS behaviors relevant to disease pathogenesis and joint damage, and identify a new transcriptomic signature regulated by this gene. These findings raise the possibility that DLX4, or one of its target genes and pathways may become a target for treatment.
成纤维样滑膜细胞（FLS）在类风湿性关节炎（RA）发病机制中起核心作用，其侵袭行为与疾病严重程度和关节损伤密切相关。然而，对FLS侵袭性的调控仍不完全清楚。远端缺失同源盒4（DLX4）是一种与癌细胞侵袭和转移有关的转录因子，我们认为它也可能参与FLS表型的调控。与对照小干扰RNA（siRNA）相比，使用siRNA敲低RA FLS中的DLX4。然后在侵袭、迁移、增殖和黏附试验中研究细胞，并使用RNA进行RNA测序和通路分析。DLX4的siRNA敲低显著降低了RA FLS的侵袭性（P = 0.028），并在划痕/伤口愈合试验中降低了迁移能力（P = 0.008）。RNA测序分析表明，DLX4敲低显著影响了与多种癌症及癌症生物学相关的基因所涉及的过程和通路，包括细胞侵袭、细胞周期、DNA复制、转录、p53信号转导、RHO GTPase信号传导、胞质分裂调控，以及对氧水平和氧化应激的反应。与对照相比，HMOX1和SOD2是DLX4敲低的FLS中表达增加最显著的基因之一，而CPNE3和HADHA是表达降低最显著的基因之一。我们描述了DLX4在调控与疾病发病机制和关节损伤相关的RA FLS行为中的新作用，并鉴定了受该基因调控的新转录组特征。这些发现增加了DLX4或其靶基因及通路之一可能成为治疗靶点的可能性。
