胰腺导管腺癌（PDAC）是所有实体瘤中最致命的癌症之一。一线治疗依赖于吉西他滨 (Gem)，尽管治疗方法有所改进，但难治性仍然是一个普遍的挑战。近年来，破译反馈环如何控制耐药信号通路的尝试引起了人们的关注，特别是磷酸酶的作用。在这项研究中，进行了基于 CRISPR/Cas9 的表型筛选，以鉴定可能对 PDAC 细胞中 Gem 反应起作用的双特异性磷酸酶 (DUSP) 家族成员。该方法揭示了非典型 RNA 磷酸酶 DUSP11 是一个潜在靶标，其抑制作用导致 PDAC 细胞对 Gem 的脆弱性。 DUSP11 基因抑制损害细胞存活并促进细胞凋亡，协同增强 Gem 细胞毒性。对 PDAC 人类样本的 RNA-seq 数据进行计算机转录组分析，发现 NF-ĸB 信号通路与 DUSP11 上调高度相关。一致地，在体外抑制 DUSP11 后，Gem 诱导的 NF-ĸB 磷酸化被阻断。从机制上讲，我们发现 PDAC 细胞暴露于 Gem 后，DUSP11 直接影响 nc886 表达并调节 PKR-NF-ĸB 信号级联，从而抵抗 Gem 诱导的细胞死亡。总之，本研究提供了有关 DUSP11 在 RNA 生物学中的作用和 PDAC 细胞中 Gem 反应的新见解。© 2024。作者获得 Springer Nature America, Inc. 的独家许可。

Pancreatic ductal adenocarcinoma (PDAC) represents one of the deadliest cancers among all solid tumors. First-line treatment relies on gemcitabine (Gem) and despite treatment improvements, refractoriness remains a universal challenge. Attempts to decipher how feedback-loops control signaling pathways towards drug resistance have gained attention in recent years, particularly focused on the role of phosphatases. In this study, a CRISPR/Cas9-based phenotypic screen was performed to identify members from the dual-specificity phosphatases (DUSP) family potentially acting on Gem response in PDAC cells. The approach revealed the atypical RNA phosphatase DUSP11 as a potential target, whose inhibition creates vulnerability of PDAC cells to Gem. DUSP11 genetic inhibition impaired cell survival and promoted apoptosis, synergistically enhancing Gem cytotoxicity. In silico transcriptome analysis of RNA-seq data from PDAC human samples identified NF-ĸB signaling pathway highly correlated with DUSP11 upregulation. Consistently, Gem-induced NF-ĸB phosphorylation was blocked upon DUSP11 inhibition in vitro. Mechanistically, we found that DUSP11 directly impacts nc886 expression and modulates PKR-NF-ĸB signaling cascade after Gem exposure in PDAC cells resulting in resistance to Gem-induced cell death. In conclusion, this study provides new insights on DUSP11 role in RNA biology and Gem response in PDAC cells.© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.