Dataset Information

Extracellular matrix and ?₅?₁ integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.

ABSTRACT: Stromal vascular fraction (SVF) cells of human adipose tissue have the capacity to generate osteogenic grafts with intrinsic vasculogenic properties. However, adipose-derived stromal/stem cells (ASC), even after minimal monolayer expansion, display poor osteogenic capacity in vivo. We investigated whether ASC bone-forming capacity may be maintained by culture within a self-produced extracellular matrix (ECM) that recapitulates the native environment. SVF cells expanded without passaging up to 28 days (Unpass-ASC) deposited a fibronectin-rich extracellular matrix and displayed greater clonogenicity and differentiation potential in vitro compared to ASC expanded only for 6 days (P0-ASC) or for 28 days with regular passaging (Pass-ASC). When implanted subcutaneously, Unpass-ASC produced bone tissue similarly to SVF cells, in contrast to P0- and Pass-ASC, which mainly formed fibrous tissue. Interestingly, clonogenic progenitors from native SVF and Unpass-ASC expressed low levels of the fibronectin receptor ?₅ integrin (CD49e), which was instead upregulated in P0- and Pass-ASC. Mechanistically, induced activation of ?₅?₁ integrin in Unpass-ASC led to a significant loss of bone formation in vivo. This study shows that ECM and regulation of ?₅?₁-integrin signaling preserve ASC progenitor properties, including bone tissue-forming capacity, during in vitro expansion.

SUBMITTER: Di Maggio N

PROVIDER: S-EPMC5349595 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Extracellular matrix and α<sub>5</sub>β<sub>1</sub> integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.

Di Maggio Nunzia N Martella Elisa E Frismantiene Agne A Resink Therese J TJ Schreiner Simone S Lucarelli Enrico E Jaquiery Claude C Schaefer Dirk J DJ Martin Ivan I Scherberich Arnaud A

Scientific reports 20170314

Stromal vascular fraction (SVF) cells of human adipose tissue have the capacity to generate osteogenic grafts with intrinsic vasculogenic properties. However, adipose-derived stromal/stem cells (ASC), even after minimal monolayer expansion, display poor osteogenic capacity in vivo. We investigated whether ASC bone-forming capacity may be maintained by culture within a self-produced extracellular matrix (ECM) that recapitulates the native environment. SVF cells expanded without passaging up to 28 ...[more]

PMID: 28290502

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Project description:Chemotherapy resistance to glioblastoma (GBM) remains an obstacle that is difficult to overcome, leading to poor prognosis of GBM patients. Many previous studies have focused on resistance mechanisms intrinsic to cancer cells; the microenvironment surrounding tumor cells has been found more recently to have significant impacts on the response to chemotherapeutic agents. Extracellular matrix (ECM) proteins may confer cell adhesion-mediated drug resistance (CAMDR). Here, expression of the ECM proteins laminin, vitronectin, and fibronectin was assessed in clinical GBM tumors using immunohistochemistry. Then, patient-derived GBM cells grown in monolayers on precoated laminin, vitronectin, or fibronectin substrates were treated with cilengitide, an integrin inhibitor, and/or carmustine, an alkylating chemotherapy. Cell adhesion and viability were quantified. Transcription factor (TF) activities were assessed over time using a bioluminescent assay in which GBM cells were transduced with lentiviruses containing consensus binding sites for specific TFs linked to expression a firefly luciferase reporter. Apoptosis, mediated by p53, was analyzed by Western blotting and immunocytofluorescence. Integrin α v activation of the FAK/paxillin/AKT signaling pathway and effects on expression of the proliferative marker Ki67 were investigated. To assess effects of integrin α v activation of AKT and ERK pathways, which are typically deregulated in GBM, and expression of epidermal growth factor receptor (EGFR), which is amplified and/or mutated in many GBM tumors, shRNA knockdown was used. Laminin, vitronectin, and fibronectin were abundant in clinical GBM tumors and promoted CAMDR in GBM cells cultured on precoated substrates. Cilengitide treatment induced cell detachment, which was most pronounced for cells cultured on vitronectin. Cilengitide treatment increased cytotoxicity of carmustine, reversing CAMDR. ECM adhesion increased activity of NFκB and decreased that of p53, leading to suppression of p53-mediated apoptosis and upregulation of multidrug resistance gene 1 (MDR1; also known as ABCB1 or P-glycoprotein). Expression of Ki67 was correlative with activation of the integrin α v -mediated FAK/paxillin/AKT signaling pathway. EGFR expression increased with integrin α v knockdown GBM cells and may represent a compensatory survival mechanism. These results indicate that ECM proteins confer CAMDR through integrin α v in GBM cells.

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Dataset Information

Extracellular matrix and ?₅?₁ integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.

Publications

Extracellular matrix and α<sub>5</sub>β<sub>1</sub> integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.

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Dataset Information

Extracellular matrix and ?5?1 integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.

Publications

Extracellular matrix and α<sub>5</sub>β<sub>1</sub> integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Extracellular matrix and ?₅?₁ integrin signaling control the maintenance of bone formation capacity by human adipose-derived stromal cells.