E2 - Other Nonviral Delivery (including lipid nanoparticles and exosomes)
322: Ultrasound-Facilitated Brain Genome Editing
Type: Oral Abstract Session
Presentation Details
Session Title: Physical Delivery and Novel Approaches
CRISPR-mediated genome editing provides new opportunities to advance molecular medicine. Efficient delivery of CRISPR elements is critical to fully realizing the potential of this technology. CRISPR delivery to the brain is especially challenging because of the blood-brain barrier (BBB), which only permits the passage of certain small molecules. Despite significant efforts to develop new adeno-associated virus (AAV) serotypes with brain tropism and brain-targeting nanoparticles, brain genome editing through systemic administration remains inefficient or limited to certain animal models. Local intracranial administration is still the gold standard for CRISPR delivery to the brain. However, for diseases that may require multiple doses, this invasive administration route is not ideal. Here we present a focused ultrasound (FUS) delivery approach that enables somatic genome editing in the brain in a noninvasive manner. When co-administered with microbubbles, the confined acoustic field can elevate BBB permeability via microbubble cavitation-induced mechanical stress, thereby enhancing penetration of CRISPR element-encoding AAV or nonviral vectors into the FUS-targeted region. In this study, we first optimized the FUS conditions for AAV delivery followed by screening of a library of DNA-barcoded nonviral polyplexes for brain genome editing. With our inhouse-optimized AAV vector, we demonstrated >25% gene editing efficiency in the target neurons in two different mouse models, while we also identified a potential polyplex composition for this application through our screening. Taken together, we confirm the feasibility of FUS for genome editing in the brain with both viral and nonviral vectors; our findings could potentially add to the armamentarium of somatic genome editing strategies for treating intractable genetic disorders in the brain.
Plain Language Summary
In this study, we demonstrate a noninvasive way for effective brain genome editing using focused ultrasound-mediated delivery of CRISPR/Cas9 genome editors. Modified from the ultrasound system seen in clinics, our focused ultrasound technique can open the blood-brain barrier for large biomolecules including genome editors to penetrate into a specific brain region. Our animal study results show a gene editing efficiency higher than 25% in the target neurons when in conjunction with a FDA-approved viral vector system, while we also demonstrate the feasibility of brain genome editing with nanoparticles.
Yeh-Hsing Lao1, Robin Ji2, Naoto Yoshinaga3, Kathy Snow4, Chun-Wei Chi1, Stephen Murray4, Elisa Konofagou2, Kam W. Leong2
1University at Buffalo, The State University of New York, Buffalo, NY,2Columbia University, New York, NY,3RIKEN, Wako, Japan,4The Jackson Laboratory, Bar Harbor, ME"
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