Targeting TNFRSF1A gene by siRNA Efficiently Blocks Extracranial Metastasis of Brain Cancer in 3D Bioprinted Tissue Model
Abstract – The Blood-Brain Barrier (BBB) is crucial for maintaining homeostasis in the brain, acting as a protective interface between the brain parenchyma and the cerebral blood vessels. The high selectiveness of the BBB renders it difficult for therapeutics to successfully cross its barrier and treat neurobiological diseases. Effective brain cancer treatment has been a challenge for scientists due to the metastatic nature of the cancer cells and the structural transformation of the BBB tissue. The TNFRSF1A gene is known to transform the BBB into the Brain-Tumor Barrier (BTB). Thus, we hypothesized that targeting the TNFRSF1A gene using siRNA would successfully downregulate its mRNA expression levels inside the blood endothelial cells. This would block the BBB-BTB transformation and consequently prevent the metastasis of the brain cancer cells across the endothelial cell barrier. We optimized the siTNFRSF1A concentration by conducting RT-PCR and agarose gel electrophoresis and investigating the effects of different concentrations on mRNA expression levels of the TNFRSF1A gene. To further confirm this, we observed the effects of siTNFRSF1A transfection of endothelial cells on brain cancer cell invasion in a 3D-printed BBB model. We found that a concentration of 1 µM of siTNFRSF1A most effectively blocked brain cancer cell penetration across the artificial BBB. Our findings suggest that treating blood endothelial cells with siTNFRSF1A may develop potential therapeutics for brain cancer patients.