title: Peptides derived from the dependence receptor ALK are proapoptotic for ALK-positive tumors.
abstract: ALK is a receptor tyrosine kinase with an oncogenic role in various types of human malignancies. Despite constitutive activation of the kinase through gene alterations, such as chromosomal translocation, gene amplification or mutation, treatments with kinase inhibitors invariably lead to the development of resistance. Aiming to develop new tools for ALK targeting, we took advantage of our previous demonstration identifying ALK as a dependence receptor, implying that in the absence of ligand the kinase-inactive ALK triggers or enhances apoptosis. Here, we synthesized peptides mimicking the proapoptotic domain of ALK and investigated their biological effects on tumor cells. We found that an ALK-derived peptide of 36 amino acids (P36) was cytotoxic for ALK-positive anaplastic large-cell lymphoma and neuroblastoma cell lines. In contrast, ALK-negative tumor cells and normal peripheral blood mononuclear cells were insensitive to P36. The cytotoxic effect was due to caspase-dependent apoptosis and required N-myristoylation of the peptide. Two P36-derived shorter peptides as well as a cyclic peptide also induced apoptosis. Surface plasmon resonance and mass spectrometry analysis of P36-interacting proteins from two responsive cell lines, Cost lymphoma and SH-SY5Y neuroblastoma, uncovered partners that could involve p53-dependent signaling and pre-mRNA splicing. Furthermore, siRNA-mediated knockdown of p53 rescued these cells from P36-induced apoptosis. Finally, we observed that a treatment combining P36 with the ALK-specific inhibitor crizotinib resulted in additive cytotoxicity. Therefore, ALK-derived peptides could represent a novel targeted therapy for ALK-positive tumors.
authors.affiliation: 1] Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France [2] Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, M5G 1X5, Canada [3] Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Aubry
authors.affiliation: 1] Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France [2] CHU Purpan, Toulouse F-31300, France.
authors.forename: S
authors.initials: S
authors.lastname: Galiacy
authors.affiliation: Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France.
authors.forename: L
authors.initials: L
authors.lastname: Ceccato
authors.affiliation: Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France.
authors.forename: C
authors.initials: C
authors.lastname: Marchand
authors.affiliation: Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France.
authors.forename: C
authors.initials: C
authors.lastname: Tricoire
authors.affiliation: INSERM, UMR1037, CRCT, Toulouse F-31000, France.
authors.forename: F
authors.initials: F
authors.lastname: Lopez
authors.affiliation: 1] Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, M5G 1X5, Canada [2] Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, M5S 1A1, Canada.
authors.forename: R
authors.initials: R
authors.lastname: Bremner
authors.affiliation: 1] INSERM, UMR 1043, CPTP, Toulouse F-31300, France [2] CNRS, UMR 5282, CPTP, Toulouse F-31300, France.
authors.forename: C
authors.initials: C
authors.lastname: Racaud-Sultan
authors.affiliation: CNRS, UMR 5089, IPBS, Toulouse F-31077, France.
authors.forename: B
authors.initials: B
authors.lastname: Monsarrat
authors.affiliation: 1] Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France [2] CHU Purpan, Toulouse F-31300, France.
authors.forename: F
authors.initials: F
authors.lastname: Malecaze
authors.affiliation: Université de Toulouse, UPS, EA4555, GR2DE, CPTP, Toulouse F-31300, France.
authors.forename: M
authors.initials: M
authors.lastname: Allouche
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D040761:Biomimetic Materials / Q000494:pharmacology; D045744:Cell Line, Tumor; D000077547:Crizotinib; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D009369:Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000473:pathology; D009447:Neuroblastoma / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D010446:Peptide Fragments / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D000077547:Crizotinib; C000626173:ALK protein, human; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases