Y Rajesh1, Anupam Banerjee2, Ipsita Pal1, Angana Biswas1, Subhayan Das1, Kaushik Kumar Dey3, Neelkamal Kapoor4, Ananta Kumar Ghosh5, Pralay Mitra6, Mahitosh Mandal7. 1. School of Medical Science and Technology, Indian Institute of Technology, Kharagpur, India. 2. Advanced Technology Development Centre, Indian Institute of Technology, Kharagpur, India. 3. School of Medical Science and Technology, Indian Institute of Technology, Kharagpur, India; Structural Biology & Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, USA. 4. Department of Pathology and Lab Medicine, All India Institute of Medical Sciences, Bhopal, India. 5. Department of Biotechnology, Indian Institute of Technology, Kharagpur, India. 6. Department of Computer Science and Engineering, Indian Institute of Technology, Kharagpur, India. 7. School of Medical Science and Technology, Indian Institute of Technology, Kharagpur, India. Electronic address: mahitosh@smst.iitkgp.ac.in.
Abstract
BACKGROUND: Epithelial to mesenchymal transition (EMT) and extracellular matrix (ECM) remodeling, are the two elemental processes promoting glioblastoma (GBM). In the present work we propose a mechanistic modelling of GBM and in process establish a hypothesis elucidating critical crosstalk between heat shock proteins (HSPs) and matrix metalloproteinases (MMPs) with synergistic upregulation of EMT-like process and ECM remodeling. METHODS: The interaction and the precise binding site between the HSP and MMP proteins was assayed computationally, in-vitro and in GBM clinical samples. RESULTS: A positive crosstalk of HSP27 with MMP-2 and MMP-9 was established in both GBM patient tissues and cell-lines. This association was found to be of prime significance for ECM remodeling and promotion of EMT-like characteristics. In-silico predictions revealed 3 plausible interaction sites of HSP27 interacting with MMP-2 and MMP-9. Site-directed mutagenesis followed by in-vitro immunoprecipitation assay (IP) with 3 mutated recombinant HSP27, confirmed an interface stretch containing residues 29-40 of HSP27 to be a common interaction site for both MMP-2 and MMP-9. This was further validated with in-vitro IP of truncated (sans AA 29-40) recombinant HSP27 with MMP-2 and MMP-9. CONCLUSION: The association of HSP27 with MMP-2 and MMP-9 proteins along with the identified interacting stretch has the potential to contribute towards drug development to inhibit GBM infiltration and migration. GENERAL SIGNIFICANCE: Current findings provide a novel therapeutic target for GBM opening a new horizon in the field of GBM management.
BACKGROUND: Epithelial to mesenchymal transition (EMT) and extracellular matrix (ECM) remodeling, are the two elemental processes promoting glioblastoma (GBM). In the present work we propose a mechanistic modelling of GBM and in process establish a hypothesis elucidating critical crosstalk between heat shock proteins (HSPs) and matrix metalloproteinases (MMPs) with synergistic upregulation of EMT-like process and ECM remodeling. METHODS: The interaction and the precise binding site between the HSP and MMP proteins was assayed computationally, in-vitro and in GBM clinical samples. RESULTS: A positive crosstalk of HSP27 with MMP-2 and MMP-9 was established in both GBM patient tissues and cell-lines. This association was found to be of prime significance for ECM remodeling and promotion of EMT-like characteristics. In-silico predictions revealed 3 plausible interaction sites of HSP27 interacting with MMP-2 and MMP-9. Site-directed mutagenesis followed by in-vitro immunoprecipitation assay (IP) with 3 mutated recombinant HSP27, confirmed an interface stretch containing residues 29-40 of HSP27 to be a common interaction site for both MMP-2 and MMP-9. This was further validated with in-vitro IP of truncated (sans AA 29-40) recombinant HSP27 with MMP-2 and MMP-9. CONCLUSION: The association of HSP27 with MMP-2 and MMP-9 proteins along with the identified interacting stretch has the potential to contribute towards drug development to inhibit GBM infiltration and migration. GENERAL SIGNIFICANCE: Current findings provide a novel therapeutic target for GBM opening a new horizon in the field of GBM management.
Authors: Gabriel Nery de Albuquerque Rego; Arielly da Hora Alves; Mariana Penteado Nucci; Javier Bustamante Mamani; Fernando Anselmo de Oliveira; Lionel Fernel Gamarra Journal: Int J Mol Sci Date: 2020-06-24 Impact factor: 5.923
Authors: Artemiy S Silantyev; Luca Falzone; Massimo Libra; Olga I Gurina; Karina Sh Kardashova; Taxiarchis K Nikolouzakis; Alexander E Nosyrev; Christopher W Sutton; Panayiotis D. Mitsias; Aristides Tsatsakis Journal: Cells Date: 2019-08-09 Impact factor: 6.600
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