Lung SORT LNPs enable precise homology-directed repair mediated CRISPR/Cas genome correction in cystic fibrosis...

Area of Science:

• Biotechnology and Genetic Engineering
• Nanomedicine
• Pulmonary Medicine

Background:

• Cystic Fibrosis (CF) affects approximately 10% of patients with CF transmembrane conductance regulator (CFTR) gene nonsense mutations, who currently lack effective treatments.
• Efficient delivery of CRISPR/Cas gene editing components to target organs remains a significant hurdle for developing gene correction therapies for CF.

Purpose of the Study:

• To develop and optimize Lung Selective Organ Targeting Lipid Nanoparticles (SORT LNPs) for efficient delivery of CRISPR/Cas components.
• To demonstrate the efficacy of gene correction therapy using SORT LNPs in preclinical models of Cystic Fibrosis.

Main Methods:

• Development of optimized Lung SORT LNPs capable of delivering Cas9 mRNA, sgRNA, and donor ssDNA templates.
• In vivo delivery of mRNA to lung basal cells in Ai9 reporter mice using optimized Lung SORT LNPs.
• Assessment of homology-directed repair-mediated gene correction in CFTR mutations in homozygous G542X mice and patient-derived bronchial epithelial cells.

Main Results:

• Optimized Lung SORT LNPs successfully delivered mRNA to target lung cells.
• SORT LNP treatment achieved precise correction of CFTR mutations in both mouse models and human cells.
• Restoration of CFTR protein expression and chloride transport function was observed post-treatment.

Conclusions:

• This study presents a proof-of-concept for using advanced SORT LNPs for CRISPR/Cas-mediated gene correction in Cystic Fibrosis.
• The developed delivery system shows significant potential for advancing mRNA LNP-based therapies for CF patients with specific genetic mutations.