Long-term correction of hemophilia B using adenoviral delivery of CRISPR/Cas9

JOURNAL OF CONTROLLED RELEASE

卷:298

Article

Stephens, Calvin J.^1,2  Lauron, Elvin J.³  Kashentseva, Elena¹  Lu, Zhi Hong¹  Yokoyama, Wayne M.³  Curiel, David T.^1,4 

[1] Washington Univ, Sch Med, Dept Radiat Oncol, Canc Biol Div, 660 South Euclid Ave,Campus Box 8224, St Louis, MO 63110 USA

[2] Washington Univ, Sch Med, Div Biol & Biomed Sci, Mol Genet & Genom Program, 660 South Euclid Ave,Campus Box 8226, St Louis, MO 63110 USA

[3] Washington Univ, Dept Med, Div Rheumatol, Sch Med, 660 South Euclid Ave,Campus Box 8045, St Louis, MO 63110 USA

[4] Washington Univ, Dept Radiat Oncol, Sch Med, Biol Therapeut Ctr, 660 South Euclid Ave,Campus Box 8224, St Louis, MO 63110 USA

关键词: Gene therapy;    Gene editing;    CRISPR/Cas9;    Hemophilia;    Adenovirus;   

DOI  :  10.1016/j.jconrel.2019.02.009

来源: Elsevier

【 摘 要 】

Hemophilia B (HB) is a life-threatening inherited disease caused by mutations in the FIX gene, leading to reduced protein function and abnormal blood clotting. Due to its monogenic nature, HB is one of the primary targets for gene therapy. Indeed, successful correction of HB has been shown in clinical trials using gene therapy approaches. However, application of these strategies to non-adult patients is limited due to high cell turnover as young patients develop, resulting in vector dilution and subsequent loss of therapeutic expression. Gene editing can potentially overcome this issue by permanently inserting the corrective gene. Integration allows replication of the therapeutic transgene at every cell division and can avoid issues associated with vector dilution. In this study, we explored adenovirus as a platform for corrective CRISPR/Cas9-mediated gene knock-in. We determined as a proof-of-principle that adenoviral delivery of CRISPR/Cas9 is capable of corrective gene addition, leading to long-term augmentation of FIX activity and phenotypic correction in a murine model of juvenile HB. While we found on-target error-free integration in all examined samples, some mice also contained mutations at the integration target site. Additionally, we detected adaptive immune responses against the vector and Cas9 nuclease. Overall, our findings show that the adenovirus platform is suitable for gene insertion in juveniles with inherited disease, suggesting this approach may be applicable to other diseases.

【 授权许可】

Free