Targeting of NLRP3 inflammasome with gene editing for the amelioration of inflammatory diseases
The NLRP3 inflammasome is a well studied target for the treatment of multiple inflammatory diseases, but how to promote the current therapeutics remains a large challenge. CRISPR/Cas9, as a gene editing tool, allows for direct ablation of NLRP3 at the genomic level. In this study, we screen an optimized cationic lipid assisted nanoparticle (CLAN) to deliver Cas9 mRNA (mCas9) and guide RNA (gRNA) into macrophages. By using CLAN encapsulating mCas9 and gRNA targeting NLRP3 (gNLRP3) (CLANmCas9/gNLRP3) we disrupt NLRP3 of macrophages, inhibiting the activation of the NLRP3 inflammasome in response to diverse stimuli. After intravenous injection, CLANmCas9/gNLRP3 mitigates acute inflammation of LPS induced septic shock and monosodium urate crystal (MSU) induced peritonitis. In addition, CLANmCas9/gNLRP3 treatment improves insulin sensitivity and reduces adipose inflammation of high fat diet (HFD) induced type 2 diabetes (T2D) Thus, our study provides a promising strategy for treating NLRP3 dependent inflammatory diseases and provides a carrier for delivering CRISPR/Cas9 into macrophages.Activation of the NLRP3 inflammasome triggers the production of inflammatory cytokines. Here, the authors inactivate NLRP3 in macrophages using CRISPR/Cas9 encapsulated in nanoparticles, and show that administration in mice is effective in preventing septic shock and peritonitis, and in improving diabetes associated inflammation and insulin resistance.
Nature Communications