title: Loss-of-function screen in rhabdomyosarcoma identifies CRKL-YES as a critical signal for tumor growth.
abstract: To identify novel signaling pathways necessary for rhabdomyosarcoma (RMS) survival, we performed a loss-of-function screen using an inducible small hairpin RNA (shRNA) library in an alveolar and an embryonal RMS cell line. This screen identified CRKL expression as necessary for growth of alveolar RMS and embryonal RMS both in vitro and in vivo. We also found that CRKL was uniformly highly expressed in both RMS cell lines and tumor tissue. As CRKL is a member of the CRK adapter protein family that contains an SH2 and two SH3 domains and is involved in signal transduction from multiple tyrosine kinase receptors, we evaluated CRKL interaction with multiple tyrosine kinase receptor signaling pathways in RMS cells. While we saw no interaction of CRKL with IGFIR, MET or PI3KAKT/mTOR pathways, we determined that CRKL signaling was associated with SRC family kinase (SFK) signaling, specifically with YES kinase. Inhibition of SFK signaling with dasatinib or another SFK inhibitor, sarcatinib, suppressed RMS cell growth in vitro and in vivo. These data identify CRKL as a novel critical component of RMS growth. This study also demonstrates the use of functional screening to identify a potentially novel therapeutic target and treatment approach for these highly aggressive pediatric cancers.
authors: Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.
authors: C L
authors: CL
authors: Yeung
authors: V N
authors: VN
authors: Ngo
authors: P J
authors: PJ
authors: Grohar
authors: F I
authors: FI
authors: Arnaldez
authors: A
authors: A
authors: Asante
authors: X
authors: X
authors: Wan
authors: J
authors: J
authors: Khan
authors: S M
authors: SM
authors: Hewitt
authors: C
authors: C
authors: Khanna
authors: L M
authors: LM
authors: Staudt
authors: L J
authors: LJ
authors: Helman
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D000069439:Dasatinib; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051575:Proto-Oncogene Proteins c-yes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011743:Pyrimidines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering; D012208:Rhabdomyosarcoma / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics; D013844:Thiazoles / Q000494:pharmacology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C082869:CRKL protein; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D034741:RNA, Small Interfering; D013844:Thiazoles; D051575:Proto-Oncogene Proteins c-yes; C489834:YES1 protein, human; D000069439:Dasatinib