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

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

doi:10.1038/s41421-023-00592-6

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

Cell Discov. 2023 Sep 15;9(1):95. doi: 10.1038/s41421-023-00592-6.

Authors

Shuyu Zhai^#^{1

2

3}, Jiewei Lin^#⁴, Yuchen Ji^#^{1

2

3}, Ronghao Zhang^#^{1

2

3}, Zehui Zhang^#^{1

2

3}, Yizhi Cao^{1

2

3}, Yang Liu^{1

2

3}, Xiaomei Tang^{1

2

3}, Jia Liu^{1

2

3}, Pengyi Liu^{1

2

3}, Jiayu Lin^{1

2

3}, Fanlu Li^{1

2

3}, Hongzhe Li^{1

2

3}, Yusheng Shi^{5

6

7}, Da Fu^{8

9

10}, Xiaxing Deng^{11

12

13}, Baiyong Shen^{14

15

16}

Affiliations

¹ Department of General Surgery, Pancreatic Disease Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
² Research Institute of Pancreatic Diseases, Shanghai Jiaotong University School of Medicine, Shanghai, China.
³ State Key Laboratory of Oncogenes and Related Genes, Institute of Translational Medicine, Shanghai Jiaotong University, Shanghai, China.
⁴ Department of Thoracic Surgery, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China.
⁵ Department of General Surgery, Pancreatic Disease Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. 815212808@qq.com.
⁶ Research Institute of Pancreatic Diseases, Shanghai Jiaotong University School of Medicine, Shanghai, China. 815212808@qq.com.
⁷ State Key Laboratory of Oncogenes and Related Genes, Institute of Translational Medicine, Shanghai Jiaotong University, Shanghai, China. 815212808@qq.com.
⁸ Department of General Surgery, Pancreatic Disease Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. fuda@shsmu.edu.cn.
⁹ Research Institute of Pancreatic Diseases, Shanghai Jiaotong University School of Medicine, Shanghai, China. fuda@shsmu.edu.cn.
¹⁰ State Key Laboratory of Oncogenes and Related Genes, Institute of Translational Medicine, Shanghai Jiaotong University, Shanghai, China. fuda@shsmu.edu.cn.
¹¹ Department of General Surgery, Pancreatic Disease Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. dxx10716@rjh.com.cn.
¹² Research Institute of Pancreatic Diseases, Shanghai Jiaotong University School of Medicine, Shanghai, China. dxx10716@rjh.com.cn.
¹³ State Key Laboratory of Oncogenes and Related Genes, Institute of Translational Medicine, Shanghai Jiaotong University, Shanghai, China. dxx10716@rjh.com.cn.
¹⁴ Department of General Surgery, Pancreatic Disease Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. shenby@shsmu.edu.cn.
¹⁵ Research Institute of Pancreatic Diseases, Shanghai Jiaotong University School of Medicine, Shanghai, China. shenby@shsmu.edu.cn.
¹⁶ State Key Laboratory of Oncogenes and Related Genes, Institute of Translational Medicine, Shanghai Jiaotong University, Shanghai, China. shenby@shsmu.edu.cn.

^# Contributed equally.

Abstract

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.

Abstract

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