title: Proteolytic fragments of fibronectin function as matrikines driving the chemotactic affinity of prostate cancer cells to human bone marrow mesenchymal stromal cells via the α5β1 integrin.
abstract: The haematopoietic niche is contributed to by bone marrow-resident mesenchymal stromal cells (BM-MSCs) and subverted by prostate cancer cells. To study mechanisms by which BM-MSCs and prostate cancer cells may interact, we assessed the migration, invasion, adhesion and proliferation of bone-derived prostate cancer cells (PC-3) in co-culture with pluripotent human BM-MSCs. We observed a strong adhesive, migratory and invasive phenotype of PC-3 cells with BM- MSC-co-culture and set out to isolate and characterize the bioactive principle. Initial studies indicated that chemotaxis was secondary to a protein residing in the >100kDa fraction. Size-exclusion chromatography (SEC) recovered peak activity in a high-molecular weight fraction containing thrombospondin-1 (TSP1). While TSP1 immunodepletion decreased activity, put-back with purified TSP1 did not reproduce bioactivity. Further purification of the TSP1-containing high-molecular weight fraction of the BM-MSC secretome with heparin-affinity chromatography recovered bioactivity with highly restricted bands on polyacrylamide gel electrophoresis, determined by mass spectroscopy to be proteolytic fragments of fibronectin (FN). Put-back experiments with full-length FN permitted adhesion but failed to induce migration. Monospecific antibodies to FN blocked adhesion. Proteolytic cleavage of FN generated FN fragments which now induced migration. Neutralizing monoclonal antibodies to FN receptors α5 and β1 integrins, and α5 knockdown specifically blocked migration and adhesion.
  
  
  
  Fibronectin fragments (FNFr) function as matrikines driving the chemotactic affinity of prostate cancer cells via the α5β1 integrin. Taken together with the high-frequency of α5β1 expression in disseminated prostate cancer cells in bone marrow aspirates from patients, the FNFr/FN-α5β1 interaction warrants further study as a therapeutic target.
authors.affiliation: a Molecular Oncology Research Institute , Tufts Medical Center , Boston , MA , USA.
authors.forename: Raghav
authors.initials: R
authors.lastname: Joshi
authors.affiliation: a Molecular Oncology Research Institute , Tufts Medical Center , Boston , MA , USA.
authors.forename: Edi
authors.initials: E
authors.lastname: Goihberg
authors.affiliation: a Molecular Oncology Research Institute , Tufts Medical Center , Boston , MA , USA.
authors.forename: Wenying
authors.initials: W
authors.lastname: Ren
authors.affiliation: b Department of Pathology , Tufts Medical Center , Boston , MA , USA.
authors.forename: Monika
authors.initials: M
authors.lastname: Pilichowska
authors.affiliation: a Molecular Oncology Research Institute , Tufts Medical Center , Boston , MA , USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Mathew
mesh_terms: D001842:Bone and Bones / Q000473:pathology; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002633:Chemotaxis; D002846:Chromatography, Affinity; D018920:Coculture Techniques; D005353:Fibronectins / Q000378:metabolism*; D006493:Heparin; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology*; D059748:Proteolysis; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D019700:Thrombospondin 1 / Q000378:metabolism
chemical_list: D005353:Fibronectins; D039081:Integrin alpha5beta1; D020543:Proteome; D019700:Thrombospondin 1; D006493:Heparin
keywords: adhesion; bone marrow mesenchymal stromal cells; bone metastases; chemotaxis; fibronectin fragments; matrikine; prostate cancer