LINC00261 编码的微蛋白 N1DARP 通过干扰 USP10-N1ICD 相互作用，促进 Notch1 细胞内领域（N1ICD）的降解，在 Notch1 高活化的胰腺癌中抑制耐药性。
A microprotein N1DARP encoded by LINC00261 promotes Notch1 intracellular domain (N1ICD) degradation via disrupting USP10-N1ICD interaction to inhibit chemoresistance in Notch1-hyperactivated pancreatic cancer.
发表日期：2023 Sep 15
作者：
Shuyu Zhai, Jiewei Lin, Yuchen Ji, Ronghao Zhang, Zehui Zhang, Yizhi Cao, Yang Liu, Xiaomei Tang, Jia Liu, Pengyi Liu, Jiayu Lin, Fanlu Li, Hongzhe Li, Yusheng Shi, Da Fu, Xiaxing Deng, Baiyong Shen
来源：
Cell Discovery
摘要：
胰腺癌细胞中显著激活的Notch信号通路在癌变、化疗抗性和复发中起重要作用。靶向该通路是胰腺癌的一个有前途的治疗策略，然而目前已报道的成功途径很少，目前使用的该通路的分子抑制剂在临床上的益处有限。在本研究中，我们鉴定了一个以前尚未表征的微小蛋白，Notch1降解相关调控多肽（N1DARP），由LINC00261编码。N1DARP敲除加速了胰腺癌器官样体和LSL-Kras、LSL-Trp53、Pdx1-Cre（KPC）小鼠中的肿瘤进展，并增强了其干细胞特性。机制上，N1DARP通过竞争性破坏N1ICD和泛素特异性肽酶10（USP10）之间的相互作用，抑制了Notch1经典和非经典通路，从而促进了N1ICD的K11和K48链聚泛素化。为评估N1DARP的治疗潜力，我们设计了一种可细胞穿透的稳定多肽SAH-mAH2-5，其螺旋结构与N1DARP相似，具有出色的物理化学稳定性。SAH-mAH2-5与N1ICD相互作用，并促进了N1ICD的蛋白酶体介导降解。在Notch1激活的胰腺癌模型中，SAH-mAH2-5注射提供了实质性的治疗效益，同时具有有限的非靶向和系统性不良反应。综上，这些发现确认了N1DARP通过调控USP10-Notch1致癌信号通路来发挥肿瘤抑制和化敏作用，并暗示了通过靶向N1DARP-N1ICD相互作用的有前途的胰腺癌治疗策略。 © 2023. 中国科学院分子细胞科学卓越中心。
The extensively activated Notch signaling pathway in pancreatic cancer cells is important in carcinogenesis, chemoresistance, and recurrence. Targeting this pathway is a promising therapeutic strategy for pancreatic cancer; however, few successful approaches have been reported, and currently used molecular inhibitors of this pathway exhibit limited clinical benefits. In this study, we identified a previously uncharacterized microprotein, Notch1 degradation-associated regulatory polypeptide (N1DARP), encoded by LINC00261. N1DARP knockout accelerated tumor progression and enhanced stem cell properties in pancreatic cancer organoids and LSL-Kras, LSL-Trp53, and Pdx1-Cre (KPC) mice. Mechanistically, N1DARP suppressed canonical and non-canonical Notch1 pathways by competitively disrupting the interaction between N1ICD and ubiquitin-specific peptidase 10 (USP10), thereby promoting K11- and K48-linked polyubiquitination of N1ICD. To evaluate the therapeutic potential of N1DARP, we designed a cell-penetrating stapled peptide, SAH-mAH2-5, with a helical structure similar to that of N1DARP that confers remarkable physicochemical stability. SAH-mAH2-5 interacted with and promoted the proteasome-mediated degradation of N1ICD. SAH-mAH2-5 injection provided substantial therapeutic benefits with limited off-target and systemic adverse effects in Notch1-activated pancreatic cancer models. Taken together, these findings confirm that N1DARP acts as a tumor suppressor and chemosensitizer by regulating USP10-Notch1 oncogenic signaling, and suggest a promising therapeutic strategy targeting the N1DARP-N1ICD interaction in Notch1-activated pancreatic cancer.© 2023. Center for Excellence in Molecular Cell Science, CAS.