Blood-brain barrier-penetrating single CRISPR-Cas9 nanocapsules for effective and safe glioblastoma gene therapy

Sci Adv. 2022 Apr 22;8(16):eabm8011. doi: 10.1126/sciadv.abm8011. Epub 2022 Apr 20.

Abstract

We designed a unique nanocapsule for efficient single CRISPR-Cas9 capsuling, noninvasive brain delivery and tumor cell targeting, demonstrating an effective and safe strategy for glioblastoma gene therapy. Our CRISPR-Cas9 nanocapsules can be simply fabricated by encapsulating the single Cas9/sgRNA complex within a glutathione-sensitive polymer shell incorporating a dual-action ligand that facilitates BBB penetration, tumor cell targeting, and Cas9/sgRNA selective release. Our encapsulating nanocapsules evidenced promising glioblastoma tissue targeting that led to high PLK1 gene editing efficiency in a brain tumor (up to 38.1%) with negligible (less than 0.5%) off-target gene editing in high-risk tissues. Treatment with nanocapsules extended median survival time (68 days versus 24 days in nonfunctional sgRNA-treated mice). Our new CRISPR-Cas9 delivery system thus addresses various delivery challenges to demonstrate safe and tumor-specific delivery of gene editing Cas9 ribonucleoprotein for improved glioblastoma treatment that may potentially be therapeutically useful in other brain diseases.

MeSH terms

  • Animals
  • Blood-Brain Barrier
  • CRISPR-Cas Systems
  • Gene Editing
  • Genetic Therapy
  • Glioblastoma* / genetics
  • Glioblastoma* / therapy
  • Mice
  • Nanocapsules*
  • RNA, Guide, Kinetoplastida / genetics

Substances

  • Nanocapsules
  • RNA, Guide