In Vivo Potency Testing of Subretinal rAAV5.hCNGB1 Gene Therapy in the Cngb1 Knockout Mouse Model of Retinitis Pigmentosa

被引：8

作者：

Wagner, Johanna E. ^{[1
]}

Zobel, Lena ^{[1
,2
]}

Gerhardt, Maximilian J. ^{[2
]}

O'Riordan, Catherine R. ^{[3
]}

Frederick, Amy ^{[3
]}

Petersen-Jones, Simon M. ^{[4
]}

Biel, Martin ^{[1
]}

Michalakis, Stylianos ^{[1
,2
]}

机构：

[1] Ludwig Maximilians Univ Munchen, Ctr Drug Res, Dept Pharm, Munich, Germany

[2] Ludwig Maximilians Univ Munchen, Univ Hosp LMU, Dept Ophthalmol, Munich, Germany

[3] Sanofi, Rare Dis, Gene Therapy, Framingham, MA USA

[4] Michigan State Univ, Vet Med Ctr, E Lansing, MI 48824 USA

来源：

HUMAN GENE THERAPY | 2021年 / 32卷 / 19-20期

基金：

美国国家卫生研究院;

关键词：

adeno-associated virus; AAV; retinal gene therapy; retinitis pigmentosa; CNGB1;

D O I：

10.1089/hum.2021.121

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Retinitis pigmentosa type 45 (RP45) is an autosomal-recessively inherited blinding disease caused by mutations in the cyclic nucleotide-gated channel subunit beta 1 (CNGB1) gene. In this study, we developed and tested a novel gene supplementation therapy suitable for clinical translation. To this end, we designed a recombinant adeno-associated virus (rAAV) vector carrying a genome that features a novel human rhodopsin promoter (hRHO194) driving rod-specific expression of full-length human CNGB1 (rAAV5.hCNGB1). rAAV5.hCNGB1 was evaluated for efficacy in the Cngb1 knockout (Cngb1(-/-)) mouse model of RP45. In particular, increasing doses of rAAV5.hCNGB1 were delivered through single subretinal injection in 4-week-old Cngb1(-/-) mice and the treatment effect was assessed over a follow-up period of 9 months at the level of (1) retinal morphology, (2) retinal function, (3) vision-guided behavior, and (4) transgene expression. We found that subretinal treatment with rAAV5.hCNGB1 resulted in efficient expression of the human CNGB1 protein in mouse rods and was able to normalize the expression of the endogenous mouse CNGA1 subunit, which together with CNGB1 forms the native heterotetrameric cyclic guanosine monophosphate-gated cation channel in rod photoreceptors. The treatment led to a dose-dependent recovery of rod photoreceptor-driven function and preservation of retinal morphology in Cngb1(-/-) mice. In summary, these results demonstrate the efficacy of hCNGB1 gene supplementation therapy in the Cngb1(-/-) mouse model of RP45 and support the translation of this approach toward future clinical application.

引用

页码：1158 / 1170

页数：13