Increased matrix metalloproteinases expression in tuberous sclerosis complex: modulation by microRNA 146a and 147b in vitro

• Published in: Neuropathology and applied neurobiology 2020-02, Vol.46 (2), p.142-159
• Main Authors: Broekaart, D. W. M., Scheppingen, J., Anink, J. J., Wierts, L., Hof, B., Jansen, F. E., Spliet, W. G., van Rijen, P. C., Kamphuis, W. W., de Vries, H. E., Aronica, E., van Vliet, E. A.
• Format: Article
• Language: English
• Subjects: Astrocytes; Blood-brain barrier; Brain - metabolism; Brain - pathology; Child, Preschool; Epilepsy; Extracellular matrix; Gelatinase A; Gelatinase B; Gene expression; Giant cells; Humans; Immunohistochemistry; Inflammation; Localization; Male; Matrix metalloproteinase; matrix metalloproteinases; Matrix Metalloproteinases - metabolism; microRNA; MicroRNAs; MicroRNAs - metabolism; miRNA; Neurodevelopmental disorders; Original; Polymerase chain reaction; Proteolysis; Stromelysin 1; Tissue inhibitor of metalloproteinase 1; Tissue inhibitor of metalloproteinase 2; Tissue inhibitor of metalloproteinase 3; Tissue inhibitor of metalloproteinase 4; Tissue Inhibitor of Metalloproteinases - metabolism; Transcription; Tuberous sclerosis; Tuberous Sclerosis - metabolism; Tuberous Sclerosis - pathology; tuberous sclerosis complex; Tumor Cells, Cultured
• ISSN: 0305-1846; 1365-2990
• DOI: 10.1111/nan.12572

Aim Matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors (TIMPs) control proteolysis within the extracellular matrix (ECM) of the brain. Dysfunction of this enzymatic system due to brain inflammation can disrupt the blood‐brain barrier (BBB) and has been implicated in the pathogenesis of epilepsy. However, this has not been extensively studied in the epileptogenic human brain. Methods We investigated the expression and cellular localization of major MMPs (MMP2, MMP3, MMP9 and MMP14) and TIMPs (TIMP1, TIMP2, TIMP3 and TIMP4) using quantitative real‐time polymerase chain reaction (RT‐PCR) and immunohistochemistry in resected epileptogenic brain tissue from patients with tuberous sclerosis complex (TSC), a severe neurodevelopmental disorder characterized by intractable epilepsy and prominent neuroinflammation. Furthermore, we determined whether anti‐inflammatory microRNAs, miR146a and miR147b, which can regulate gene expression at the transcriptional level, could attenuate dysregulated MMP and TIMP expression in TSC tuber‐derived astroglial cultures. Results We demonstrated higher mRNA and protein expression of MMPs and TIMPs in TSC tubers compared to control and perituberal brain tissue, particularly in dysmorphic neurons and giant cells, as well as in reactive astrocytes, which was associated with BBB dysfunction. More importantly, IL‐1β‐induced dysregulation of MMP3, TIMP2, TIMP3 and TIMP4 could be rescued by miR146a and miR147b in tuber‐derived TSC cultures. Conclusions This study provides evidence of dysregulation of the MMP/TIMP proteolytic system in TSC, which is associated with BBB dysfunction. As dysregulated MMP and TIMP expression can be ameliorated in vitro by miR146a and miR147b, these miRNAs deserve further investigation as a novel therapeutic approach.