2. Academic Validation
  3. Disrupting YAP1-mediated glutamine metabolism induces synthetic lethality alongside ODC1 inhibition in osteosarcoma

Disrupting YAP1-mediated glutamine metabolism induces synthetic lethality alongside ODC1 inhibition in osteosarcoma

  • Cell Oncol (Dordr). 2024 Aug 8. doi: 10.1007/s13402-024-00967-1.
Hongsheng Wang # 1 2,  Yining Tao # 1 2,  Jing Han # 1 2,  Jiakang Shen # 1 2,  Haoran Mu 1 2,  Zhuoying Wang 1 2,  Jinzeng Wang 3,  Xinmeng Jin 1 2,  Qi Zhang 4,  Yuqin Yang 5,  Jun Lin 6,  Mengxiong Sun 1 2,  Xiaojun Ma 1 2,  Ling Ren 7,  Amy K LeBlanc 7,  Jing Xu 8 9,  Yingqi Hua 10 11,  Wei Sun 12 13
Affiliations

Affiliations

  • 1 Department of Orthopedics, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, 100 Haining Road, Shanghai, 200080, China.
  • 2 Shanghai Bone Tumor Institution, Shanghai, China.
  • 3 National Research Center for Translational Medicine (Shanghai), State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
  • 4 Department of Biochemistry and Molecular Cell Biology, Shanghai Key Laboratory for Tumor Microenvironment and Inflammation, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
  • 5 Department of Laboratory Animal Center, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
  • 6 Department of Pathology, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
  • 7 Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
  • 8 Department of Orthopedics, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, 100 Haining Road, Shanghai, 200080, China. jingxu6000@163.com.
  • 9 Shanghai Bone Tumor Institution, Shanghai, China. jingxu6000@163.com.
  • 10 Department of Orthopedics, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, 100 Haining Road, Shanghai, 200080, China. yhua@shsmu.edu.cn.
  • 11 Shanghai Bone Tumor Institution, Shanghai, China. yhua@shsmu.edu.cn.
  • 12 Department of Orthopedics, Shanghai General Hospital, School of Medicine, Shanghai Jiao Tong University, 100 Haining Road, Shanghai, 200080, China. viv-sun@sjtu.edu.cn.
  • 13 Shanghai Bone Tumor Institution, Shanghai, China. viv-sun@sjtu.edu.cn.
  • # Contributed equally.
PMID: 39115605 DOI: 10.1007/s13402-024-00967-1
Abstract

Purpose: Osteosarcoma, a highly malignant primary bone tumor primarily affecting adolescents, frequently develops resistance to initial chemotherapy, leading to metastasis and limited treatment options. Our study aims to uncover novel therapeutic targets for metastatic and recurrent osteosarcoma.

Methods: In this study, we proved the potential of modulating the YAP1-regulated glutamine metabolic pathway to augment the response of OS to DFMO. We initially employed single-cell transcriptomic data to gauge the activation level of polyamine metabolism in MTAP-deleted OS patients. This was further substantiated by transcriptome Sequencing data from recurrent and non-recurrent patient tissues, confirming the activation of polyamine metabolism in progressive OS. Through high-throughput drug screening, we pinpointed CIL56, a YAP1 inhibitor, as a promising candidate for a combined therapeutic strategy with DFMO. In vivo, we utilized PDX and CDX models to validate the therapeutic efficacy of this drug combination. In vitro, we conducted western blot analysis, qPCR analysis, immunofluorescence staining, and PuMA experiments to monitor alterations in molecular expression, distribution, and tumor metastasis capability. We employed CCK-8 and colony formation assays to assess the proliferative capacity of cells in the experimental group. We used flow cytometry and reactive oxygen probes to observe changes in ROS and glutamine metabolism within the cells. Finally, we applied RNA-seq in tandem with metabolomics to identify metabolic alterations in OS cells treated with a DFMO and CIL56 combination. This enabled us to intervene and validate the role of the YAP1-mediated glutamine metabolic pathway in DFMO resistance.

Results: Through single-cell RNA-seq data analysis, we pinpointed a subset of late-stage OS cells with significantly upregulated polyamine metabolism. This upregulation was further substantiated by transcriptomic profiling of recurrent and non-recurrent OS tissues. High-throughput drug screening revealed a promising combination strategy involving DFMO and CIL56. DFMO treatment curbs the phosphorylation of YAP1 protein in OS cells, promoting nuclear entry and initiating the YAP1-mediated glutamine metabolic pathway. This reduces intracellular ROS levels, countering DFMO's Anticancer effect. The therapeutic efficacy of DFMO can be amplified both in vivo and in vitro by combining it with the YAP1 inhibitor CIL56 or the Glutaminase Inhibitor CB-839. This underscores the significant potential of targeting the YAP1-mediated glutamine metabolic pathway to enhance efficacy of DFMO.

Conclusion: Our findings elucidate YAP1-mediated glutamine metabolism as a crucial bypass mechanism against DFMO, following the inhibition of polyamine metabolism. Our study provides valuable insights into the potential role of DFMO in an "One-two Punch" therapy of metastatic and recurrent osteosarcoma.

Keywords

Glutamine metabolism; High-throughput drug screening; Osteosarcoma; Polyamine metabolism; Single-cell RNA-seq; YAP1.

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