Elena Rampazzo1, Monica Dettin2, Francesca Maule3, Alessandra Scabello3, Luisa Calvanese4, Gabriella D'Auria4, Lucia Falcigno4, Elena Porcù3, Annj Zamuner2, Alessandro Della Puppa5, Daniele Boso3, Giuseppe Basso3, Luca Persano6. 1. Department of Woman and Children's Health, University of Padova, via Giustiniani 3, 35128 Padova, Italy; Istituto di Ricerca Pediatrica (IRP) Città della Speranza, corso Stati Uniti 4, 35127 Padova, Italy. 2. Department of Industrial Engineering, University of Padova, via Marzolo 9, 35128 Padova, Italy. 3. Department of Woman and Children's Health, University of Padova, via Giustiniani 3, 35128 Padova, Italy. 4. Department of Pharmacy, University Federico II of Naples, via Domenico Montesano 49, 80131 Naples, Italy; CIRPeB, University Federico II of Naples, via Mezzocannone 16, 80134 Naples, Italy. 5. Neurosurgery Unit, University-Hospital of Padova, via Giustiniani 2, 35128 Padova, Italy. 6. Istituto di Ricerca Pediatrica (IRP) Città della Speranza, corso Stati Uniti 4, 35127 Padova, Italy. Electronic address: luca.persano@unipd.it.
Abstract
BACKGROUND: Glioblastoma (GBM) is the most aggressive type of primary brain tumor, characterized by the intrinsic resistance to chemotherapy due to the presence of a highly aggressive Cancer Stem Cell (CSC) sub-population. In this context, Bone Morphogenetic Proteins (BMPs) have been demonstrated to induce CSC differentiation and to sensitize GBM cells to treatments. METHODS: The BMP-2 mimicking peptide, named GBMP1a, was synthesized on solid-phase by Fmoc chemistry. Structural characterization and prediction of receptor binding were obtained by Circular Dicroism (CD) and NRM analyses. Activation of BMP signalling was evaluated by a luciferase reporter assay and western blot. Pro-differentiating effects of GBMP1a were verified by immunostaining and neurosphere assay in primary glioblastoma cultures. RESULTS: CD and NMR showed that GBMP1a correctly folds into expected tridimensional structures and predicted its binding to BMPR-IA to the same epitope as in the native complex. Reporter analysis disclosed that GBMP1a is able to activate BMP signalling in GBM cells. Moreover, BMP-signalling activation was specifically dependent on smad1/5/8 phosphorylation. Finally, we confirmed that GBMP1a treatment is sufficient to enhance osteogenic differentiation of Mesenchymal Stem Cells and to induce astroglial differentiation of glioma stem cells (GSCs) in vitro. CONCLUSIONS: GBMP1a was demonstrated to be a good inducer of GSC differentiation, thus being considered a potential anti-cancer tool to be further developed for GBM treatment. GENERAL SIGNIFICANCE: These data highlight the role of BMP-mimicking peptides as potential anti-cancer agents against GBM and stimulate the further development of GBMP1a-based structures in order to enhance its stability and activity.
BACKGROUND:Glioblastoma (GBM) is the most aggressive type of primary brain tumor, characterized by the intrinsic resistance to chemotherapy due to the presence of a highly aggressive Cancer Stem Cell (CSC) sub-population. In this context, Bone Morphogenetic Proteins (BMPs) have been demonstrated to induce CSC differentiation and to sensitize GBM cells to treatments. METHODS: The BMP-2 mimicking peptide, named GBMP1a, was synthesized on solid-phase by Fmoc chemistry. Structural characterization and prediction of receptor binding were obtained by Circular Dicroism (CD) and NRM analyses. Activation of BMP signalling was evaluated by a luciferase reporter assay and western blot. Pro-differentiating effects of GBMP1a were verified by immunostaining and neurosphere assay in primary glioblastoma cultures. RESULTS: CD and NMR showed that GBMP1a correctly folds into expected tridimensional structures and predicted its binding to BMPR-IA to the same epitope as in the native complex. Reporter analysis disclosed that GBMP1a is able to activate BMP signalling in GBM cells. Moreover, BMP-signalling activation was specifically dependent on smad1/5/8 phosphorylation. Finally, we confirmed that GBMP1a treatment is sufficient to enhance osteogenic differentiation of Mesenchymal Stem Cells and to induce astroglial differentiation of glioma stem cells (GSCs) in vitro. CONCLUSIONS: GBMP1a was demonstrated to be a good inducer of GSC differentiation, thus being considered a potential anti-cancer tool to be further developed for GBM treatment. GENERAL SIGNIFICANCE: These data highlight the role of BMP-mimicking peptides as potential anti-cancer agents against GBM and stimulate the further development of GBMP1a-based structures in order to enhance its stability and activity.
Authors: Paolo Rosa; Sofia Scibetta; Giuseppe Pepe; Giorgio Mangino; Luca Capocci; Sam J Moons; Thomas J Boltje; Francesco Fazi; Vincenzo Petrozza; Alba Di Pardo; Vittorio Maglione; Antonella Calogero Journal: Int J Mol Sci Date: 2022-08-24 Impact factor: 6.208