AAV5 Delivery of CRISPR/Cas9 Mediates Genome Editing in the Lungs of Young Rhesus Monkeys

Shun-Qing Liang; Andrew W Navia; Michelle Ramseier; Xuntao Zhou; Michele Martinez; Charles Lee; Chen Zhou; Joae Wu; Jun Xie; Qin Su; Dan Wang; Terence R Flotte; Daniel G Anderson; Alice F Tarantal; Alex K Shalek; Guangping Gao; Wen Xue

doi:10.1089/hum.2024.035

AAV5 Delivery of CRISPR/Cas9 Mediates Genome Editing in the Lungs of Young Rhesus Monkeys

Hum Gene Ther. 2024 Jul 3. doi: 10.1089/hum.2024.035. Online ahead of print.

Authors

Shun-Qing Liang¹, Andrew W Navia^{2

3

4}, Michelle Ramseier^{2

3

4}, Xuntao Zhou¹, Michele Martinez⁵, Charles Lee⁵, Chen Zhou^{6

7}, Joae Wu⁸, Jun Xie^{6

7

9}, Qin Su^{6

7

9}, Dan Wang^{6

7}, Terence R Flotte⁶, Daniel G Anderson^{10

11

12

13}, Alice F Tarantal⁵, Alex K Shalek^{2

3

4}, Guangping Gao^{6

7

9}, Wen Xue^{1

7}

Affiliations

¹ RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
² Institute for Medical Engineering and Science, Department of Chemistry, and Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
³ Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
⁴ Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts, USA.
⁵ Departments of Pediatrics and Cell Biology and Human Anatomy, School of Medicine, and California National Primate Research Center, University of California, Davis, California, USA.
⁶ Horae Gene Therapy Center and Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
⁷ Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
⁸ Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
⁹ Viral Vector Core, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
¹⁰ Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
¹¹ David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
¹² Harvard and MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
¹³ Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

PMID: 38767512
DOI: 10.1089/hum.2024.035

Abstract

Genome editing has the potential to treat genetic diseases in a variety of tissues, including the lung. We have previously developed and validated a dual adeno-associated virus (AAV) CRISPR platform that supports effective editing in the airways of mice. To validate this delivery vehicle in a large animal model, we have shown that intratracheal instillation of CRISPR/Cas9 in AAV5 can edit a housekeeping gene or a disease-related gene in the lungs of young rhesus monkeys. We observed up to 8% editing of angiotensin-converting enzyme 2 (ACE2) in lung lobes after single-dose administration. Single-nuclear RNA sequencing revealed that AAV5 transduces multiple cell types in the caudal lung lobes, including alveolar cells, macrophages, fibroblasts, endothelial cells, and B cells. These results demonstrate that AAV5 is efficient in the delivery of CRISPR/Cas9 in the lung lobes of young rhesus monkeys.

Keywords: AAV5; CRISPR; lung; rhesus monkey.

Abstract

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