Silencing Adamts2 attenuates fibroblast-mediated fibrosis and promotes axonal regeneration in an in vitro model.

Fibrotic scars formed after central nervous system injury pose a strong barrier to axonal regeneration. To attenuate the inhibitory effect of fibrotic scars, numerous pre-clinical studies have investigated strategies. Fibroblasts are the main cells involved in the formation of fibrotic scars. In this study, we first used single-cell sequencing data to analyze the changes in fibroblasts after mouse spinal cord injury and screened the specifically highly expressed gene Adamts2 (metallopeptidase with thrombospondin type 1 motif 2). Subsequently, we evaluated the efficacy of Adamts2-targeting RNAi in attenuating the pro-fibrotic phenotype of fibroblasts using an in vitro TGFβ-induced fibroblast model. We found that TGFβ enhanced the expression of Adamts2 in primary spinal cord fibroblasts and regulated the expression of fibrosis-related genes. Moreover, silencing of Adamts2 attenuated the pro-fibrotic activity of TGFβ in spinal cord fibroblasts. Mechanistically, the knockdown of Adamts2 in fibroblasts leads to the upregulation of multiple neurotrophic factors, subsequently activating the AKT and ERK signaling pathways in motor neurons to alleviate inhibitory effects on axonogenesis. Our results demonstrate that Adamts2-specific siRNA significantly suppresses the TGFβ-induced pro-fibrotic phenotype and alleviates its inhibitory effects on motor neuron axonogenesis during co-culture. Collectively, these results indicate that inhibiting Adamts2 effectively suppresses fibroblast-mediated fibrosis, suggesting that targeting Adamts2 is a promising therapeutic strategy for promoting neural repair following spinal cord injury by promoting a neuro-supportive microenvironment.