摘要
Neuropathic pain affects 3-10% of the worldwide population, but current therapies are inadequate to provide sustained pain relief. Inflammation plays key role in neuropathic pain, and anti-inflammatory interleukin 10 (IL-10) represses inflammation and pain. Cre/loxP system is a genetic tool that enables site-specific DNA recombination. Here, we developed a hybrid Cre/loxP-based baculovirus system to deliver RNA-guided CRISPR activation (CRISPRa) to trigger IL-10 expression in adipose-derived stem cells (ASC), hoping to synergize the immunosuppressive effects of ASC and IL-10 to mitigate inflammation and attenuate neuropathic pain. We constructed a hybrid baculovirus system for CRISPRa delivery into ASC to activate IL-10. We showed that rat ASC does not naturally express IL-10 but can be activated by baculovirus-mediated CRISPRa delivery to specifically induce prolonged IL-10 secretion. The extended IL-10 expression in ASC promoted macrophage polarization from pro-inflammatory M1-like towards anti-inflammatory M2-like phenotype under inflammatory conditions, which simultaneously suppressed pro-inflammatory tumor necrosis factor α (TNF-α)/ interleukin 6 (IL-6) and upregulated anti-inflammatory IL-10/transforming growth factor β1 (TGF-β1). The IL-10 activation in ASC improved the dorsal root ganglion (DRG) neuron outgrowth, repressed the hypersensitivity of DRG neurons in vitro, and prevented the mechanical/cold allodynia and heat hyperalgesia starting from day 7 and persisted for at least 28 days. RNA-guided activation system to trigger IL-10 expression in ASC thus represents a new durable therapeutic approach for neuropathic pain relief. Yang et al. developed a hybrid baculovirus system for CRISPRa delivery into ASC to activate and extended IL-10 expression. The extended IL-10 expression in ASC switched macrophage polarization, suppressed inflammation, repressed neuron hypersensitivity and attenuated pain for 28 days, representing a new durable therapeutic approach for neuropathic pain relief.