한빛사논문
- 조회325
Hye Yeon Choi 1,10, Yicheng Zhu 1,10, Xuyao Zhao 1, Simran Mehta 1, Juan Carlos Hernandez 1, Jae-Jin Lee 1, Yi Kou 2, Risa Machida 1, Mauro Giacca 3, Giannino Del Sal 4,5,6, Ratna Ray 7, Hyungjin Eoh 1, Stanley M. Tahara 1, Lin Chen 2, Hidekazu Tsukamoto 8,9 and Keigo Machida 1,9,*
1Departments of Molecular Microbiology and Immunology, University of Southern California, Los Angeles, CA, USA.
2Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA.
3International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
4Department of Life Sciences, University of Trieste, 34127 Trieste, Italy.
5International Centre for Genetic Engineering and Biotechnology (ICGEB), Area Science Park-Padriciano, Trieste, Italy.
6IFOM ETS, The AIRC Institute of Molecular Oncology, Milan, Italy.
7Saint Louis University, School of Medicine, St Louis, MO, USA.
8Department of Pathology, University of Southern California, Los Angeles, CA, USA.
9Southern California Research Center for ALPD and Cirrhosis, Los Angeles, CA, USA.
10These authors contributed equally: Hye Yeon Choi, Yicheng Zhu.
*Corresponding author: correspondence to Keigo Machida
Abstract
The P53-destabilizing TBC1D15-NOTCH protein interaction promotes self-renewal of tumor-initiating stem-like cells (TICs); however, the mechanisms governing the regulation of this pathway have not been fully elucidated. Here, we show that TBC1D15 stabilizes NOTCH and c-JUN through blockade of E3 ligase and CDK8 recruitment to phosphodegron sequences. Chromatin immunoprecipitation (ChIP-seq) analysis was performed to determine whether TBC1D15-dependent NOTCH1 binding occurs in TICs or non-TICs. The TIC population was isolated to evaluate TBC1D15-dependent NOTCH1 stabilization mechanisms. The tumor incidence in hepatocyte-specific triple knockout (Alb::CreERT2;Tbc1d15Flox/Flox;Notch1Flox/Flox;Notch2Flox/Flox;HCV-NS5A) Transgenic (Tg) mice and wild-type mice was compared after being fed an alcohol-containing Western diet (WD) for 12 months. The NOTCH1-TBC1D15-FIS1 interaction resulted in recruitment of mitochondria to the perinuclear region. TBC1D15 bound to full-length NUMB and to NUMB isoform 5, which lacks three Ser phosphorylation sites, and relocalized NUMB5 to mitochondria. TBC1D15 binding to NOTCH1 blocked CDK8- and CDK19-mediated phosphorylation of the NOTCH1 PEST phosphodegron to block FBW7 recruitment to Thr-2512 of NOTCH1. ChIP-seq analysis revealed that TBC1D15 and NOTCH1 regulated the expression of genes involved in mitochondrial metabolism-related pathways required for the maintenance of TICs. TBC1D15 inhibited CDK8-mediated phosphorylation to stabilize NOTCH1 and protect it from degradation The NUMB-binding oncoprotein TBC1D15 rescued NOTCH1 from NUMB-mediated ubiquitin-dependent degradation and recruited NOTCH1 to the mitochondrial outer membrane for the generation and expansion of liver TICs. A NOTCH-TBC1D15 inhibitor was found to inhibit NOTCH-dependent pathways and exhibited potent therapeutic effects in PDX mouse models. This unique targeting of the NOTCH-TBC1D15 interaction not only normalized the perinuclear localization of mitochondria but also promoted potent cytotoxic effects against TICs to eradicate patient-derived xenografts through NOTCH-dependent pathways.
논문정보
- Research article
- 2024년 02월 (BRIC 등록일 2024-03-29)
- 국외 연구진
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