F Bussolino

Università degli Studi di Torino, Torino, Piedmont, Italy

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Publications (328)1745.2 Total impact

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    ABSTRACT: The mechanism by which angiogenic endothelial cells break the physical barrier of the vascular basement membrane and consequently sprout to form new vessels in mature tissues is unclear. Here, we show that the angiogenic endothelium is characterized by the presence of functional podosome rosettes. These extracellular-matrix-degrading and adhesive structures are precursors of de novo branching points and represent a key feature in the formation of new blood vessels. VEGF-A stimulation induces the formation of endothelial podosome rosettes by upregulating integrin α6β1. In contrast, the binding of α6β1 integrin to the laminin of the vascular basement membrane impairs the formation of podosome rosettes by restricting α6β1 integrin to focal adhesions and hampering its translocation to podosomes. Using an ex vivo sprouting angiogenesis assay, transgenic and knockout mouse models and human tumour sample analysis, we provide evidence that endothelial podosome rosettes control blood vessel branching and are critical regulators of pathological angiogenesis.
    Nature Cell Biology 09/2014; · 20.76 Impact Factor
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    ABSTRACT: During organogenesis, patterning is primarily achieved by the combined actions of morphogens. Among these, semaphorins represent a general system for establishing the appropriate wiring architecture of biological nets. Originally discovered as evolutionarily conserved steering molecules for developing axons, subsequent studies on semaphorins expanded their functions to the cardiovascular and immune systems. Semaphorins participate in cardiac organogenesis and control physiological vasculogenesis and angiogenesis, which result from a balance between pro- and anti-angiogenic signals. These signals are altered in several diseases. In this review, we discuss the role of semaphorins in vascular biology, emphasizing the mechanisms by which these molecules control vascular patterning and lymphangiogenesis, as well as in genetically inherited and degenerative vascular diseases.
    Trends in Molecular Medicine 08/2014; 10(2):1-10. · 9.57 Impact Factor
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    ABSTRACT: Directional cell migration is of paramount importance in both physiological and pathological processes, such as development, wound healing, immune response, and cancer invasion. Here, we report that 3-phosphoinositide-dependent kinase 1 (PDK1) regulates epithelial directional migration and invasion by binding and activating myotonic dystrophy kinase-related CDC42-binding kinase α (MRCKα). We show that the effect of PDK1 on cell migration does not involve its kinase activity but instead relies on its ability to bind membrane phosphatidylinositol (3,4,5)-trisphosphate. Upon epidermal growth factor (EGF) stimulation, PDK1 and MRCKα colocalize at the cell membrane in lamellipodia. We demonstrate that PDK1 positively modulates MRCKα activity and drives its localization within lamellipodia. Likewise, the retraction phase of lamellipodia is controlled by PDK1 through an MRCKα-dependent mechanism. In summary, we discovered a functional pathway involving PDK1-mediated activation of MRCKα, which links EGF signaling to myosin contraction and directional migration.
    The Journal of Cell Biology 08/2014; 206(3):415-34. · 10.82 Impact Factor
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    ABSTRACT: Angiopoietin-like (ANGPTL) proteins are secreted proteins showing structural similarity to members of the angiopoietin family. Some ANGPTL proteins possess pleiotropic activities, being involved in cancer lipid, glucose energy metabolisms, and angiogenesis. ANGPTL7 is the less characterized member of the family whose functional role is only marginally known. In this study, we provide experimental evidences that ANGPTL7 is over-expressed in different human cancers. To understand the role played by ANGPTL7 in tumor biology, we asked whether ANGPTL7 is endogenously expressed by malignant cells or in response to environmental stimuli. We found that ANGPTL7 is marginally expressed under standard growth condition while it is specifically up-regulated by hypoxia. Interestingly, the protein is secreted and partially associated with the exosomal fraction, suggesting that it could be found in the systemic circulation of oncologic patients and act in an endocrine way. Moreover, we found that ANGPTL7 exerts a pro-angiogenetic effect on human differentiated endothelial cells by stimulating their proliferation, motility, invasiveness, and capability to form capillary-like networks while it does not stimulate progenitor endothelial cells. Finally, we showed that ANGPTL7 promotes vascularization in vivo in the mouse Matrigel sponge assay, thereby accrediting this molecule as a pro-angiogenic factor.
    Angiogenesis 06/2014; · 4.41 Impact Factor
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    ABSTRACT: The synaptic protein Neuroligin 1 (NLGN1), a cell adhesion molecule, is critical for the formation and consolidation of synaptic connectivity and is involved in vascular development. The mechanism through which NLGN1 acts, especially in vascular cells, is unknown. Here, we aimed at deepening our knowledge on the cellular activities and molecular pathways exploited by endothelial NLGN1 both in vitro and in vivo. We analyzed the phenotypic consequences of NLGN1 expression modulation in Endothelial cells (ECs) through in vitro angiogenesis assays and the mouse postnatal retinal angiogenesis model. We demonstrate that NLGN1, while not affecting ECs proliferation or migration, modulates cell adhesion to the vessel stabilizing protein laminin through cooperation with the α6 integrin, a specific laminin receptor. Finally, we show that in vivo, NLGN1 and α6 integrin preferentially colocalize in the mature retinal vessels, while NLGN1 deletion causes an aberrant VE-cadherin, laminin and α6 integrin distribution in vessels, along with significant structural defects in the vascular tree
    Journal of Biological Chemistry 05/2014; · 4.65 Impact Factor
  • Federico Bussolino, Enrico Giraudo, Guido Serini
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    ABSTRACT: Semaphorins were originally identified as axon guidance molecules involved in the development of the neuronal system. However, accumulating evidences have clearly demonstrated that the semaphorin system is not restricted to the brain but supports functions of other organs. Here, we review the rapidly emerging functions of sempahorins and, in particular class 3 semaphorin, in vascular and lymphatic systems during the development, tumor angiogenesis and ischemic revascularization.
    Clinical Reviews in Allergy & Immunology 01/2014; 99:71-88. · 5.59 Impact Factor
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    ABSTRACT: Supplementary Figure for "Diffusion limited phase separation in eukaryotic chemotaxis", P.N.A.S. 2005, 102, 16927
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    ABSTRACT: Supplementary Text for "Diffusion limited phase separation in eukaryotic chemotaxis", P.N.A.S. 2005, 102, 16927
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    ABSTRACT: Supplementary Movie for "Diffusion limited phase separation in eukaryotic chemotaxis", P.N.A.S. 2005, 102, 16927
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    ABSTRACT: Preventing cell entry of human immunodeficiency virus 1 (HIV-1) is of interest for the development of innovative therapies. We previously reported a specific interaction between HIV-1 envelope glycoprotein 120 (gp120) and Tat at the cell surface, which enhances virus attachment and entry. We also identified a gp120-mimicking peptide, CT319, that competes with gp120 for Tat binding, thus inhibiting HIV-1 infection. Here we report a molecular dissection of gp120 regions involved in this mechanism. Our findings identify the V1/V2 loop of gp120 as involved in Tat binding, and define this interaction as functionally relevant for HIV-1 entry into host cells. Tatphysically interactswithgp120bypull down(1,2,3,4).
    FEBS letters 07/2013; · 3.54 Impact Factor
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    ABSTRACT: Molecular targeting of drug delivery nanocarriers is expected to improve their therapeutic index while decreasing their toxicity. Here we report the identification and characterization of novel peptide ligands specific for cells present in high-risk neuroblastoma (NB), a childhood tumor mostly refractory to current therapies. To isolate such targeting moieties, we performed combined in vitro/ex-vivo phage display screenings on NB cell lines and on tumors derived from orthotopic mouse models of human NB. By designing proper subtractive protocols, we identified phage clones specific either for the primary tumor, its metastases, or for their respective stromal components. Globally, we isolated 121 phage-displayed NB-binding peptides: 26 bound the primary tumor, 15 the metastatic mass, 57 and 23 their respective microenvironments. Of these, five phage clones were further validated for their specific binding ex-vivo to biopsies from stage IV NB patients and to NB tumors derived from mice. All five clones also targeted tumor cells and vasculature in vivo when injected into NB-bearing mice. Coupling of the corresponding targeting peptides with doxorubicin-loaded liposomes led to a significant inhibition in tumor volume and enhanced survival in preclinical NB models, thereby paving the way to their clinical development.
    Journal of Controlled Release 05/2013; · 7.63 Impact Factor
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    ABSTRACT: The intrinsic complexity of the process of vessel formation limits the efficacy of cellular assays for elucidation of its molecular and pharmacological mechanisms. We developed an ex vivo three-dimensional assay of sprouting angiogenesis with arterial explants from human umbilical cords. In this assay, human arterial rings were embedded in basement membrane extract gel, leading to a network of capillary-like structures upon VEGF-A stimulation. The angiogenic outgrowth consisted of endothelial cells, which actively internalized acetylated-LDL, surrounded by pericytes. Computer-assisted quantification of this vascular network demonstrated considerable sensitivity of this assay to several angiogenic inhibitors, including kinase inhibitors and monoclonal antibodies. We also performed targeted gene knock-down on this model by directly infecting explanted umbilical arteries with lentiviruses carrying short-hairpin RNA. Down-regulation of VEGFR2 resulted in a significant reduction of the sprouting capability, demonstrating the relevance of human vascular explants for functional genomics studies. Furthermore, a modification of this assay led to development of a three-dimensional model of tumor-driven angiogenesis, in which angiogenic outgrowth was sustained by spheroids of prostate cancer cells in absence of exogenous growth factors. The human arterial ring assay bridges the gap between in vitro endothelial cell and animal model, and is a powerful system for identification of genes and drugs that regulate human angiogenesis.
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    ABSTRACT: Key points Human Arterial Ring assay is an innovative system for the three-dimensional study of tumor angiogenesisThis assay can be exploited for anti-angiogenic drug screening and gene function analysis on human vessels.
    Blood 03/2013; · 9.78 Impact Factor
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    ABSTRACT: The intrinsic complexity of the process of vessel formation limits the efficacy of cellular assays for elucidation of its molecular and pharmacological mechanisms. We developed an ex vivo three-dimensional assay of sprouting angiogenesis with arterial explants from human umbilical cords. In this assay, human arterial rings were embedded in basement membrane extract gel, leading to a network of capillary-like structures upon VEGF-A stimulation. The angiogenic outgrowth consisted of endothelial cells, which actively internalized acetylated-LDL, surrounded by pericytes. Computer-assisted quantification of this vascular network demonstrated considerable sensitivity of this assay to several angiogenic inhibitors, including kinase inhibitors and monoclonal antibodies. We also performed targeted gene knock-down on this model by directly infecting explanted umbilical arteries with lentiviruses carrying short-hairpin RNA. Down-regulation of VEGFR2 resulted in a significant reduction of the sprouting capability, demonstrating the relevance of human vascular explants for functional genomics studies. Furthermore, a modification of this assay led to development of a three-dimensional model of tumor-driven angiogenesis, in which angiogenic outgrowth was sustained by spheroids of prostate cancer cells in absence of exogenous growth factors. The human arterial ring assay bridges the gap between in vitro endothelial cell and animal model, and is a powerful system for identification of genes and drugs that regulate human angiogenesis.
    Blood 03/2013; · 9.78 Impact Factor
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    ABSTRACT: Neurexin (NRXN) and Neuroligin (NLGN) are trans-synaptic proteins involved in vascular biology. NRXN is encoded in long (α) and short (β) isoforms. We have shown that βNRXN modulates blood vessel development in synergy with VEGFA and associates with NLGN. On the other hand αNRXN is also expressed in blood vessels but does not interact with NLGN or act in synergy with VEGFA, thus demonstrating a differential role. To find clues into the vascular functions of αNRXN, we chose a 7 aa motif that is part of its extracellular region and was formerly selected through a proteomic search for interactors of the vascular receptor Tie2. Next we a) synthetized and modeled such peptide in order to determine its biological activity towards Tie2 in in vitro and in vivo angiogenesis assays and b) evaluated if αNRXN and Tie2 physically associate in situ during vascular development. We used biochemical and cellular assays to prove that the synthetic αNRXN peptide a) modulates the angiogenic phenotype of cultured endothelial cells and angiogenesis in vivo and b) efficiently stimulates Tie2 phosphorylation and downstream mediators in endothelial cells. Moreover, we show that αNRXN and Tie2 can be reciprocally immunoprecipated from chicken blood vessels at late stages of vascular development. These data have a double significance, i.e. provide a novel tool to modulate Tie2 and further suggest the involvement of the NRXN family of synaptic protein in the vascular system through their interaction with a fundamental vascular player.
    Biochemical and Biophysical Research Communications 02/2013; · 2.28 Impact Factor
  • Serena Marchiò, Elena Astanina, Federico Bussolino
    The EMBO Journal 01/2013; · 9.82 Impact Factor
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    ABSTRACT: Neurexins (Nrxs) have emerged as potential determinants of synaptic specificity, but little is known about their localization at central synapses. Here we show that Nrxs have a remarkably selective localization at distinct types of glutamatergic synapses and we reveal an unexpected ontogenetic regulation of Nrx expression at GABAergic synapses. Our data indicate that synapses are specified by molecular interactions that involve both Nrx-dependent and Nrx-independent mechanisms. We propose that differences in the spatio-temporal profile of Nrx expression may contribute to specify the molecular identity of synapses.
    Frontiers in Cellular Neuroscience 01/2013; 7:35. · 4.47 Impact Factor
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    ABSTRACT: A one-dimensional photonic crystal (1DPC) based on a planar stack of dielectric layers is used as an optical transducer for biosensing, upon the coupling of TE-polarized Bloch Surface Waves (BSW). The structure is tailored with a polymeric layer providing a chemical functionality facilitating the covalent binding of orienting proteins needed for a subsequent grafting of antibodies in an immunoassay detection scheme. The polymeric layer is impregnated with Cy3 dye, in such a way that the photonic structure can exhibit an emissive behavior. The BSW-coupled fluorescence shift is used as a means for detecting refractive index variations occurring at the 1DPC surface, according to a label-free concept. The proposed working principle is successfully demonstrated in real-time tracking of protein G covalent binding on the 1DPC surface within a fluidic cell.
    Sensors 01/2013; 13(2):2011-22. · 2.05 Impact Factor
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Publication Stats

12k Citations
1,745.20 Total Impact Points

Institutions

  • 1980–2014
    • Università degli Studi di Torino
      • • Dipartimento di Scienze Chirurgiche
      • • Dipartimento di Scienze Mediche
      • • Dipartimento di Scienze Cliniche e Biologiche
      Torino, Piedmont, Italy
  • 1998–2013
    • Politecnico di Torino
      • DISAT - Department of Applied Science and Technology
      Torino, Piedmont, Italy
  • 1999–2012
    • Institute for Cancer Research and Treatment
      • Medical Oncology Division
      Torino, Piedmont, Italy
  • 2007
    • University of Naples Federico II
      • Department of Physical Sciences
      Napoli, Campania, Italy
    • Max Planck Society
      München, Bavaria, Germany
  • 2005
    • San Raffaele Scientific Institute
      Milano, Lombardy, Italy
  • 1995–2005
    • University of Pavia
      Ticinum, Lombardy, Italy
    • Universität Erfurt
      Erfurt, Thuringia, Germany
  • 2002
    • Università degli Studi di Bari Aldo Moro
      • Dipartimento di Scienze Biomediche ed Oncologia Umana (DIMO)
      Bari, Apulia, Italy
  • 2000
    • University of Amsterdam
      Amsterdamo, North Holland, Netherlands
  • 1988–1997
    • Mario Negri Institute for Pharmacological Research
      Milano, Lombardy, Italy
  • 1987–1997
    • Polo d'Innovazione di Genomica Genetica e Biologia
      Perugia, Umbria, Italy
    • University at Buffalo, The State University of New York
      • Department of Medicine
      Buffalo, NY, United States
  • 1996
    • University of Buenos Aires
      • Biological Chemistry Department
      Buenos Aires, Buenos Aires F.D., Argentina
  • 1993–1994
    • Università degli Studi di Messina
      • Dipartimento di Medicina Clinica e Sperimentale
      Messina, Sicily, Italy
  • 1991
    • Albany State University
      Georgia, United States
  • 1987–1988
    • École Polytechnique Fédérale de Lausanne
      • Laboratoire de chimie et biochimie computationnelles
      Lausanne, VD, Switzerland