title: Autocrine Signaling by Wnt-5a Deregulates Chemotaxis of Leukemic Cells and Predicts Clinical Outcome in Chronic Lymphocytic Leukemia.
abstract: OBJECTIVE
  ROR1, a receptor in the noncanonical Wnt/planar cell polarity (PCP) pathway, is upregulated in malignant B cells of chronic lymphocytic leukemia (CLL) patients. It has been shown that the Wnt/PCP pathway drives pathogenesis of CLL, but which factors activate the ROR1 and PCP pathway in CLL cells remains unclear.
  
  
  METHODS
  B lymphocytes from the peripheral blood of CLL patients were negatively separated using RosetteSep (StemCell) and gradient density centrifugation. Relative expression of WNT5A, WNT5B, and ROR1 was assessed by quantitative real-time PCR. Protein levels, protein interaction, and downstream signaling were analyzed by immunoprecipitation and Western blotting. Migration capacity of primary CLL cells was analyzed by the Transwell migration assay.
  
  
  RESULTS
  By analyzing the expression in 137 previously untreated CLL patients, we demonstrate that WNT5A and WNT5B genes show dramatically (five orders of magnitude) varying expression in CLL cells. High WNT5A and WNT5B expression strongly associates with unmutated IGHV and shortened time to first treatment. In addition, WNT5A levels associate, independent of IGHV status, with the clinically worst CLL subgroups characterized by dysfunctional p53 and mutated SF3B1. We provide functional evidence that WNT5A-positive primary CLL cells have increased motility and attenuated chemotaxis toward CXCL12 and CCL19 that can be overcome by inhibitors of Wnt/PCP signaling.
  
  
  CONCLUSIONS
  These observations identify Wnt-5a as the crucial regulator of ROR1 activity in CLL and suggest that the autocrine Wnt-5a signaling pathway allows CLL cells to overcome natural microenvironmental regulation.
authors.affiliation: Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Pavlina
authors.initials: P
authors.lastname: Janovska
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic. CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Poppova
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic. CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Karla
authors.initials: K
authors.lastname: Plevova
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic. CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Plesingerova
authors.affiliation: Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Behal
authors.affiliation: Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Marketa
authors.initials: M
authors.lastname: Kaucka
authors.affiliation: Institute of Biostatistics and Analyses, Masaryk University, Brno, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Ovesna
authors.affiliation: CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Hlozkova
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Borsky
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic.
authors.forename: Olga
authors.initials: O
authors.lastname: Stehlikova
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic.
authors.forename: Yvona
authors.initials: Y
authors.lastname: Brychtova
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic. CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Michael
authors.initials: M
authors.lastname: Doubek
authors.affiliation: Department of Paediatric Otorhinolaryngology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Machalova
authors.affiliation: Genetics Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland.
authors.forename: Sivasubramanian
authors.initials: S
authors.lastname: Baskar
authors.affiliation: Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic.
authors.forename: Alois
authors.initials: A
authors.lastname: Kozubik
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic. CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Sarka
authors.initials: S
authors.lastname: Pospisilova
authors.affiliation: Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hematology and Oncology, University Hospital Brno and Medical Faculty, Masaryk University, Brno, Czech Republic. CEITEC-Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
authors.forename: Sarka
authors.initials: S
authors.lastname: Pavlova
authors.affiliation: Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. bryja@sci.muni.cz.
authors.forename: Vitezslav
authors.initials: V
authors.lastname: Bryja
mesh_terms: D019898:Autocrine Communication / Q000502:physiology*; D001402:B-Lymphocytes / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology; D002633:Chemotaxis / Q000502:physiology*; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D057050:Receptor Tyrosine Kinase-like Orphan Receptors / Q000378:metabolism; D015854:Up-Regulation / Q000502:physiology; D051153:Wnt Proteins / Q000378:metabolism*; D060449:Wnt Signaling Pathway / Q000502:physiology*; D000071818:Wnt-5a Protein
chemical_list: D011518:Proto-Oncogene Proteins; C085389:WNT5A protein, human; C435831:WNT5B protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C078716:ROR1 protein, human; D057050:Receptor Tyrosine Kinase-like Orphan Receptors