VE-PTP-targeted RBC-hybrid lipid nanoparticles restore endothelial ANG1–Tie2 signaling in diabetic kidney disease

Targeted delivery to injured glomerular endothelial cells remains a major challenge in diabetic kidney disease (DKD) because systemically administered therapeutics are limited by rapid kidney clearance, complex glomerular vascular architecture, and limited endothelial retention. Biomimetic red blood cell (RBC) membrane-hybrid lipid nanoparticles offer a promising strategy to improve vascular compatibility, prolong circulation, and protect therapeutic cargos. However, the application of this biomimetic platform for glomerular endothelial cell-targeted delivery of dCas9-based CRISPR activation (CRISPRa) ribonucleoprotein (RNP) complexes has not been established. dCas9-sgRNA RNP complexes were efficiently incorporated into RBC-hybrid lipid nanoparticles through charge-mediated assembly using an ionizable lipid. The resulting VE-PTP-targeted RBC/LNP-Ab nanoparticles showed stable nanoscale size, favorable surface charge modulation, efficient RNP loading, and structural features consistent with RBC membrane-hybrid nanoparticle formation. In VE-PTP-upregulated endothelial cells under DKD-mimicking conditions, the nanoparticles enhanced intracellular dCas9 delivery and increased endogenous ANG1 expression, restoring the ANG1/ANG2 balance and downstream Tie2-Akt signaling. Following systemic administration in advanced DKD mice, VE-PTP-targeted RBC-hybrid nanoparticles preferentially localized to glomerular endothelial cells, increased ANG1 expression in the diabetic kidney, reduced albuminuria, and attenuated structural kidney injury. These results demonstrate that VE-PTP-targeted RBC-hybrid nanoparticles can serve as a biomimetic non-viral platform for glomerular endothelial cell-targeted CRISPRa RNP delivery. This strategy provides proof-of-concept for disease-specific transcriptional gene modulation in DKD and may provide a framework for targeted endothelial therapy in other microvascular diseases.