Engineered CRISPR/Cas13d Virus-Like Particles for Potent, Safe Gene Therapy of Idiopathic Pulmonary Fibrosis
Idiopathic pulmonary fibrosis is a fatal, progressive lung disease that severely impacts patient's quality of life and life expectancy. Currently, effective treatment options are lacking, emphasizing the urgent need for novel therapeutic strategies. CRISPR is a promising tool for the disease gene therapy. However, viral vector delivery of CRISPR Cas13d exacerbates its adverse effects (e.g., off-target and bystander effects), which compromises the safety of in vivo disease gene therapy. Here, we report the first establishment and optimization of a CRISPR Cas13d virus-like particle (VLP) delivery system. This system delivers CRISPR Cas13d into cells as protein and RNA forms, enabling transient in vivo retention and improved safety. By employing CRISPR Cas13d VLPs to simultaneously knock down the Tgfb1, Ctgf, and Nox4 genes in a murine model of pulmonary fibrosis, we observed a robust therapeutic effect without inducing marked adverse reactions. Collectively, this study may serve as a foundational platform for advancing gene therapy in pulmonary fibrosis.