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Vav1 is a crucial molecule in monocytic/macrophagic differentiation of myeloid leukemia-derived cells

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Abstract

Vav1 is a critical signal transducer for both the development and function of normal hematopoietic cells, in which it regulates the acquisition of maturation-related properties, including adhesion, motility, and phagocytosis. Vav1 is also important for the agonist-induced maturation of acute promyelocytic leukemia (APL)-derived promyelocytes, in which it promotes the acquisition of a mature phenotype by playing multiple functions at both cytoplasmic and nuclear levels. We investigated the possible role of Vav1 in the differentiation of leukemic precursors to monocytes/macrophages. Tumoral promyelocytes in which Vav1 was negatively modulated were induced to differentiate into monocytes/macrophages with phorbol-12-myristate-13-acetate (PMA) and monitored for their maturation-related properties. We found that Vav1 was crucial for the phenotypical differentiation of tumoral myeloid precursors to monocytes/macrophages, in terms of CD11b expression, adhesion capability and cell morphology. Confocal analysis revealed that Vav1 may synergize with actin in modulating nuclear morphology of PMA-treated adherent cells. Our data indicate that, in tumoral promyelocytes, Vav1 is a component of lineage-specific transduction machineries that can be recruited by various differentiating agents. Since Vav1 plays a central role in the completion of the differentiation program of leukemic promyelocytes along diverse hematopoietic lineages, it can be considered a common target for developing new therapeutic strategies for the various subtypes of myeloid leukemias.

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Abbreviations

ATRA:

All trans-retinoic acid

APL:

Acute promyelocytic leukemia

BSA:

Bovine serum albumin

DAPI:

4,6-Diamino-2-phenylindole

DABCO:

1,4-Diazabicyclo [2.2.2] octane

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

GEF:

Guanine Nucleotide exchange factor

PBS:

Phosphate-buffered saline

PIC:

Piceatannol

PMA:

Phorbol-12-myristate-13-acetate

siRNA:

Small interfering RNA

TRITC:

Tetramethylrhodamine isothiocyanate

TBS:

TRIS-buffered saline

References

  • Bertagnolo V, Marchisio M, Brugnoli F, Bavelloni A, Boccafogli L, Colamussi ML, Capitani S (2001) Requirement of tyrosine-phosphorylated Vav for morphological differentiation of all-trans-retinoic acid-treated HL-60 cells. Cell Growth Differ 12:193–200

    PubMed  CAS  Google Scholar 

  • Bertagnolo V, Brugnoli F, Marchisio M, Celeghini C, Carini C, Capitani S (2004) Association of PI3-K with tyrosine phosphorylated Vav is essential for its activity in neutrophil-like maturation of myeloid cells. Cell Signal 16:423–433

    Article  PubMed  CAS  Google Scholar 

  • Bertagnolo V, Brugnoli F, Mischiati C, Sereni A, Bavelloni A, Carini C, Capitani S (2005) Vav promotes differentiation of human tumoral myeloid precursors. Exp Cell Res 306:56–63

    Article  PubMed  CAS  Google Scholar 

  • Bertagnolo V, Grassilli S, Bavelloni A, Brugnoli F, Piazzi M, Candiano G, Petretto A, Benedusi M, Capitani S (2008) Vav1 modulates protein expression during ATRA-induced maturation of APL-derived promyelocytes: a proteomic-based analysis. J Proteome Res 7:3729–3736

    Article  PubMed  CAS  Google Scholar 

  • Bertagnolo V, Grassilli S, D'Aguanno S, Brugnoli F, Bavelloni A, Faenza I, Nika E, Urbani A, Cocco L, Capitani S (2010) Mass spectrometry-based identification of Y745 of Vav1 as a tyrosine residue crucial in maturation of acute promyelocytic leukemia-derived cells. J Proteome Res 9:752–760

    Article  PubMed  CAS  Google Scholar 

  • Bhavsar PJ, Vigorito E, Turner M, Ridley AJ (2009) Vav GEFs regulate macrophage morphology and adhesion-induced Rac and Rho activation. Exp Cell Res 15:3345–3358

    Article  Google Scholar 

  • Brugnoli F, Lambertini E, Varin-Blank N, Piva R, Marchisio M, Grassilli S, Miscia S, Capitani S, Bertagnolo V (2010) Vav1 and PU.1 are recruited to the CD11b promoter in APL-derived promyelocytes: role of Vav1 in modulating PU.1-containing complexes during ATRA-induced differentiation. Exp Cell Res 1:38–47

    Article  Google Scholar 

  • Bustelo XR (2001) Vav proteins, adaptors and cell signaling. Oncogene 20:6372–6381

    Article  PubMed  CAS  Google Scholar 

  • Cougoule C, Hoshino S, Dart A, Lim J, Caron E (2006) Dissociation of recruitment and activation of the small G-protein Rac during Fcgamma receptor-mediated phagocytosis. J Biol Chem 281:8756–8764

    Article  PubMed  CAS  Google Scholar 

  • Denkinger DJ, Lambrecht TQ, Cushman-Vokoun AM, Kawahara RS (2002) PU.1 regulates the expression of the vav proto-oncogene. J Cell Biochem 84:772–783

    Article  PubMed  Google Scholar 

  • Fujikawa K, Miletic AV, Alt FW, Faccio R, Brown T, Hoog J, Fredericks J, Nishi S, Mildiner S, Moores SL, Brugge J, Rosen FS, Swat W (2003) Vav1/2/3-null mice define an essential role for Vav family proteins in lymphocyte development and activation but a differential requirement in MAPK signaling in T and B cells. J Exp Med 17:1595–1608

    Article  Google Scholar 

  • Gakidis MA, Cullere X, Olson T, Wilsbacher JL, Zhang B, Moores SL, Ley K, Swat W, Mayadas T, Brugge JS (2004) Vav GEFs are required for beta2 integrin-dependent functions of neutrophils. J Cell Biol 166:273–282

    Article  PubMed  CAS  Google Scholar 

  • Hall AB, Gakidis MA, Glogauer M, Wilsbacher JL, Gao S, Swat W, Brugge JS (2006) Requirements for Vav guanine nucleotide exchange factors and Rho GTPases in FcgammaR- and complement-mediated phagocytosis. Immunity 24:305–316

    Article  PubMed  CAS  Google Scholar 

  • Haubert D, Weckbecker G (2010) Vav1 couples the T cell receptor to cAMP response element activation via a PKC-dependent pathway. Cell Signal 22:944–954

    Article  PubMed  CAS  Google Scholar 

  • Holsinger LJ, Graef IA, Swat W, Chi T, Bautista DM, Davidson L, Lewis RS, Alt FW, Crabtree GR (1998) Defects in actin-cap formation in Vav-deficient mice implicate an actin requirement for lymphocyte signal transduction. Curr Biol 8:563–572

    Article  PubMed  CAS  Google Scholar 

  • Houlard M, Arudchandran R, Regnier-Ricard F, Germani A, Gisselbrecht S, Blank U, Rivera J, Varin-Blank N (2002) Vav1 is a component of transcriptionally active complexes. J Exp Med 195:1115–1127

    Article  PubMed  CAS  Google Scholar 

  • Jasek E, Mirecka J, Litwin JA (2008) Effect of differentiating agents (all-trans retinoic acid and phorbol 12-myristate 13-acetate) on drug sensitivity of HL60 and NB4 cells in vitro. Folia Histochem Cytobiol 46:323–330

    Article  PubMed  CAS  Google Scholar 

  • Katzav S (2009) Vav1: a hematopoietic signal transduction molecule involved in human malignancies. Int J Biochem Cell Biol 41:1245–1248

    Article  PubMed  CAS  Google Scholar 

  • Khatau SB, Hale CM, Stewart-Hutchinson PJ, Patel MS, Stewart CL, Searson PC, Hodzic D, Wirtz D (2009) A perinuclear actin cap regulates nuclear shape. Proc Natl Acad Sci USA 106:19017–19122

    Article  PubMed  CAS  Google Scholar 

  • Kim C, Christophe CM, Josef P, Mary CD (2003) The hemopoietic Rho/Rac guanine nucleotide exchange factor Vav1 regulates N-formyl-methionyl-leucyl-phenylalanine-activated neutrophil functions. J Immunol 171:4425–4430

    PubMed  CAS  Google Scholar 

  • Kogan SC (2009) Curing APL: differentiation or destruction? Cancer Cell 15:7–8

    Article  PubMed  CAS  Google Scholar 

  • Miletic AV, Graham DB, Sakata-Sogawa K, Hiroshima M, Hamann MJ, Cemerski S, Kloeppel T, Billadeau DD, Kanagawa O, Tokunaga M, Swat W (2009) Vav links the T cell antigen receptor to the actin cytoskeleton and T cell activation independently of intrinsic guanine nucleotide exchange activity. PLoS ONE 4:e6599

    Article  PubMed  Google Scholar 

  • Murao S, Gemmell MA, Callaham MF, Anderson NL, Huberman E (1983) Control of macrophage cell differentiation in human promyelocytic HL-60 leukemia cells by 1,25-dihydroxyvitamin D3 and phorbol-12-myristate-13-acetate. Cancer Res 43:4989–4996

    PubMed  CAS  Google Scholar 

  • Papadopulos F, Spinelli M, Valente S, Foroni L, Orrico C, Alviano F, Pasquinelli G (2007) Common tasks in microscopic and ultrastructural image analysis using ImageJ. Ultrastruct Pathol 31:401–407

    Article  PubMed  Google Scholar 

  • Park MH, Park SY, Kim Y (2008) Induction of proline-rich tyrosine kinase2 (Pyk2) through C/EBPbeta is involved in PMA-induced monocyte differentiation. FEBS Lett 582:415–422

    Article  PubMed  CAS  Google Scholar 

  • Phillipson M, Heit B, Parsons SA, Petri B, Mullaly SC, Colarusso P, Gower RM, Neely G, Simon SI, Kubes P (2009) Vav1 is essential for mechanotactic crawling and migration of neutrophils out of the inflamed microvasculature. J Immunol 182:6870–6878

    Article  PubMed  CAS  Google Scholar 

  • Qin S, Yamamura H (1997) Up-regulation of Syk activity during HL60 cell differentiation into granulocyte but not into monocyte/macrophage-lineage. Biochem Biophys Res Commun 236:697–701

    Article  PubMed  CAS  Google Scholar 

  • Shi Y, Tohyama Y, Kadono T, He J, Miah SM, Hazama R, Tanaka C, Tohyama K, Yamamura H (2006) Protein-tyrosine kinase Syk is required for pathogen engulfment in complement-mediated phagocytosis. Blood 107:4554–4562

    Article  PubMed  CAS  Google Scholar 

  • Song X, Norman AW (1998) 1Alpha,25-dihydroxyvitamin D3 and phorbol ester mediate the expression of alkaline phosphatase in NB4 acute promyelocytic leukemia cells. Leuk Res 22:69–76

    Article  PubMed  CAS  Google Scholar 

  • Stricker J, Falzone T, Gardel ML (2010) Mechanics of the F-actin cytoskeleton. J Biomech 43:9–14

    Article  PubMed  Google Scholar 

  • Tybulewicz VL (2005) Vav-family proteins in T-cell signalling. Curr Opin Immunol 17:267–274

    Article  PubMed  CAS  Google Scholar 

  • Wells CM, Bhavsar PJ, Evans IR, Vigorito E, Turner M, Tybulewicz V, Ridley AJ (2005) Vav1 and Vav2 play different roles in macrophage migration and cytoskeletal organization. Exp Cell Res 1:303–310

    Article  Google Scholar 

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Authors and Affiliations

  1. Signal Transduction Unit, Section of Human Anatomy, Department of Morphology and Embryology, University of Ferrara, Via Fossato di Mortara 66, 44100, Ferrara, Italy

    Valeria Bertagnolo, Ervin Nika, Federica Brugnoli, Silvia Grassilli & Silvano Capitani

  2. Section of General Pathology, Department of Experimental and Diagnostic Medicine, University of Ferrara, 44100, Ferrara, Italy

    Massimo Bonora & Paolo Pinton

  3. Interdisciplinary Center for the Study of Inflammation (ICSI), University of Ferrara, 44100, Ferrara, Italy

    Massimo Bonora, Paolo Pinton & Silvano Capitani

  4. Emilia Romagna Laboratory, BioPharmaNet, University of Ferrara, 44100, Ferrara, Italy

    Massimo Bonora & Paolo Pinton

  5. Laboratory for advanced technologies and therapies (LTTA), Emilia Romagna Technopole, University of Ferrara, 44100, Ferrara, Italy

    Valeria Bertagnolo, Federica Brugnoli, Paolo Pinton & Silvano Capitani

Authors
  1. Valeria Bertagnolo
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  2. Ervin Nika
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  3. Federica Brugnoli
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  4. Massimo Bonora
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  5. Silvia Grassilli
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  6. Paolo Pinton
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  7. Silvano Capitani
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Corresponding author

Correspondence to Valeria Bertagnolo.

Additional information

This research was supported by grants from MIUR Cofin (2007) to S.C., MAE (Italy-Croatia bilateral project 2009–2010) to V.B., and local funds from the University of Ferrara (Italy) to S.C. and V.B.

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Bertagnolo, V., Nika, E., Brugnoli, F. et al. Vav1 is a crucial molecule in monocytic/macrophagic differentiation of myeloid leukemia-derived cells. Cell Tissue Res 345, 163–175 (2011). https://doi.org/10.1007/s00441-011-1195-5

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  • DOI: https://doi.org/10.1007/s00441-011-1195-5

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