title: Survey of activated FLT3 signaling in leukemia.
abstract: Activating mutations of FMS-like tyrosine kinase-3 (FLT3) are found in approximately 30% of patients with acute myeloid leukemia (AML). FLT3 is therefore an attractive drug target. However, the molecular mechanisms by which FLT3 mutations lead to cell transformation in AML remain unclear. To develop a better understanding of FLT3 signaling as well as its downstream effectors, we performed detailed phosphoproteomic analysis of FLT3 signaling in human leukemia cells. We identified over 1000 tyrosine phosphorylation sites from about 750 proteins in both AML (wild type and mutant FLT3) and B cell acute lymphoblastic leukemia (normal and amplification of FLT3) cell lines. Furthermore, using stable isotope labeling by amino acids in cell culture (SILAC), we were able to quantified over 400 phosphorylation sites (pTyr, pSer, and pThr) that were responsive to FLT3 inhibition in FLT3 driven human leukemia cell lines. We also extended this phosphoproteomic analysis on bone marrow from primary AML patient samples, and identify over 200 tyrosine and 800 serine/threonine phosphorylation sites in vivo. This study showed that oncogenic FLT3 regulates proteins involving diverse cellular processes and affects multiple signaling pathways in human leukemia that we previously appreciated, such as Fc epsilon RI-mediated signaling, BCR, and CD40 signaling pathways. It provides a valuable resource for investigation of oncogenic FLT3 signaling in human leukemia.
authors.affiliation: Cell Signaling Technology, Inc., Danvers, Massachusetts, United States of America. tgu@cellsignal.com
authors.forename: Ting-lei
authors.initials: TL
authors.lastname: Gu
authors.forename: Julie
authors.initials: J
authors.lastname: Nardone
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.forename: Marc
authors.initials: M
authors.lastname: Loriaux
authors.forename: Judit
authors.initials: J
authors.lastname: Villén
authors.forename: Sean
authors.initials: S
authors.lastname: Beausoleil
authors.forename: Meghan
authors.initials: M
authors.lastname: Tucker
authors.forename: Jon
authors.initials: J
authors.lastname: Kornhauser
authors.forename: Jianmin
authors.initials: J
authors.lastname: Ren
authors.forename: Joan
authors.initials: J
authors.lastname: MacNeill
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Druker
authors.forename: Michael C
authors.initials: MC
authors.lastname: Heinrich
authors.forename: John
authors.initials: J
authors.lastname: Rush
authors.forename: Roberto D
authors.initials: RD
authors.lastname: Polakiewicz
mesh_terms: D001854:Bone Marrow Cells / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000473:pathology*; D009154:Mutation; D019606:Phosphoamino Acids / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010879:Piperazines / Q000494:pharmacology; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology*; D011799:Quinazolines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics / Q000378:metabolism*
chemical_list: D019606:Phosphoamino Acids; D010879:Piperazines; D011799:Quinazolines; C464670:tandutinib; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3