School of Medical Science and Technology, Indian Institute of Technology, Kharagpur, India.
Advanced Technology Development Centre, Indian Institute of Technology, Kharagpur, India.
Biochim Biophys Acta Gen Subj. 2019 Jul;1863(7):1196-1209. doi: 10.1016/j.bbagen.2019.04.015. Epub 2019 Apr 25.
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.
The interaction and the precise binding site between the HSP and MMP proteins was assayed computationally, in-vitro and in GBM clinical samples.
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.
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.
Current findings provide a novel therapeutic target for GBM opening a new horizon in the field of GBM management.
上皮间质转化（EMT）和细胞外基质（ECM）重塑是促进胶质母细胞瘤（GBM）的两个基本过程。在本工作中，我们提出了一个 GBM 的机制模型，并在此过程中提出了一个假设，阐明了热休克蛋白（HSPs）和基质金属蛋白酶（MMPs）之间的关键串扰，以及 EMT 样过程和 ECM 重塑的协同上调。
通过计算、体外和 GBM 临床样本检测，研究 HSP 和 MMP 蛋白之间的相互作用和精确结合位点。
在 GBM 患者组织和细胞系中均证实 HSP27 与 MMP-2 和 MMP-9 之间存在正性串扰。这种关联对 ECM 重塑和 EMT 样特征的促进具有重要意义。计算机预测揭示了 HSP27 与 MMP-2 和 MMP-9 相互作用的 3 个可能的相互作用位点。通过对 3 个突变重组 HSP27 进行定点突变和体外免疫沉淀试验（IP），证实 HSP27 的 29-40 个残基包含一个界面延伸，是 MMP-2 和 MMP-9 的共同相互作用位点。这进一步通过 HSP27 截断（不含 AA 29-40）与 MMP-2 和 MMP-9 的体外 IP 得到验证。
HSP27 与 MMP-2 和 MMP-9 蛋白的关联以及鉴定出的相互作用片段有可能为抑制 GBM 浸润和迁移的药物开发做出贡献。
目前的研究结果为 GBM 提供了一个新的治疗靶点，为 GBM 管理领域开辟了新的前景。
