title: Phosphoproteomics reveals ALK promote cell progress via RAS/ JNK pathway in neuroblastoma.
abstract: Emerging evidence suggests receptor tyrosine kinase ALK as a promising therapeutic target in neuroblastoma. However, clinical trials reveal that a limited proportion of ALK-positive neuroblastoma patients experience clinical benefits from Crizotinib, a clinically approved specific inhibitor of ALK. The precise molecular mechanisms of aberrant ALK activity in neuroblastoma remain elusive, limiting the clinical application of ALK as a therapeutic target in neuroblastoma. Here, we describe a deep quantitative phosphoproteomic approach in which Crizotinib-treated neuroblastoma cell lines bearing aberrant ALK are used to investigate downstream regulated phosphoproteins. We identified more than 19,500-and quantitatively analyzed approximately 10,000-phosphorylation sites from each cell line, ultimately detecting 450-790 significantly-regulated phosphorylation sites. Multiple layers of bioinformatic analysis of the significantly-regulated phosphoproteins identified RAS/JNK as a downstream signaling pathway of ALK, independent of the ALK variant present. Further experiments demonstrated that ALK/JNK signaling could be inactivated by either ALK- or JNK-specific inhibitors, resulting in cell growth inhibition by induction of cell cycle arrest and cell apoptosis. Our study broadly defines the phosphoproteome in response to ALK inhibition and provides a resource for further clinical investigation of ALK as therapeutic target for the treatment of neuroblastoma.
authors.affiliation: Department of Pediatric Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.|Division of Pediatric Oncology, Shanghai Institute of Pediatric Research, Shanghai, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Chen
authors.affiliation: Department of Pediatric Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.|Division of Pediatric Oncology, Shanghai Institute of Pediatric Research, Shanghai, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Lv
authors.affiliation: Department of Pediatric Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Guofeng
authors.initials: G
authors.lastname: Xu
authors.affiliation: Department of Pediatric Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.|Division of Pediatric Oncology, Shanghai Institute of Pediatric Research, Shanghai, China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Pediatric Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.|Division of Pediatric Oncology, Shanghai Institute of Pediatric Research, Shanghai, China.
authors.forename: Yeming
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Pediatric Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China.|Division of Pediatric Oncology, Shanghai Institute of Pediatric Research, Shanghai, China.
authors.forename: Zhixiang
authors.initials: Z
authors.lastname: Wu
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D019295:Computational Biology / Q000379:methods; D018450:Disease Progression; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D010750:Phosphoproteins / Q000378:metabolism*; D047428:Protein Kinase Inhibitors; D020543:Proteome; D040901:Proteomics / Q000379:methods; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D015398:Signal Transduction; D018631:ras Proteins / Q000378:metabolism*
chemical_list: D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D020543:Proteome; C000626173:ALK protein, human; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D018631:ras Proteins
keywords: ALK; neuroblastoma; pediatric oncology; phosphoproteomics; target therapy