Cas9-specific immune responses compromise local and systemic AAV CRISPR therapy in multiple dystrophic canine models
Abstract
Adeno-associated virus (AAV)-mediated CRISPR-Cas9 editing holds promise to treat many diseases. The immune response to bacterial-derived Cas9 has been speculated as a hurdle for AAV-CRISPR therapy. However, immunological consequences of AAV-mediated Cas9 expression have thus far not been thoroughly investigated in large mammals. We evaluate Cas9-specific immune responses in canine models of Duchenne muscular dystrophy (DMD) following intramuscular and intravenous AAV-CRISPR therapy. Treatment results initially in robust dystrophin restoration in affected dogs but also induces muscle inflammation, and Cas9-specific humoral and cytotoxic T-lymphocyte (CTL) responses that are not prevented by the muscle-specific promoter and transient prednisolone immune suppression. In normal dogs, AAV-mediated Cas9 expression induces similar, though milder, immune responses. In contrast, other therapeutic (micro-dystrophin and SERCA2a) and reporter (alkaline phosphatase, AP) vectors result in persistent expression without inducing muscle inflammation. Our results suggest Cas9 immunity may represent a critical barrier for AAV-CRISPR therapy in large mammals.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Nov 24, 2021
- Source ID
- 10.1038/s41467-021-26830-7
Entities
People
- Bruce F Smith
- Chady H. Hakim
- Charles A. Gersbach
- Christopher E Nelson
- Dennis O. Pérez-lópez
- Dirk Grimm
- Dong Zhang
- Dongsheng Duan
- Emily D. Million
- Feng Feng
- Florian Schmidt
- Gang Yao
- Jacqueline A. Louderman
- James Teixeira
- Jin Han
- Keqing Zhang
- N. Nora Yang
- Nalinda B. Wasala
- Roland W Herzog
- Samantha Metzger
- Sandeep R. P. Kumar
- Shi-Jie Chen
- Xiufang Pan
- Yongping Yue
Organizations
- National Institute of Arthritis and Musculoskeletal and Skin Diseases
- National Institute of General Medical Sciences
- National Institute of Neurological Disorders and Stroke
- Parent Project Muscular Dystrophy
- United States Department of Defense
- University of Missouri