AAV-CRB2 protects against vision loss in an inducible CRB1 retinitis pigmentosa mouse model

TY - JOUR

T1 - AAV-CRB2 protects against vision loss in an inducible CRB1 retinitis pigmentosa mouse model

AU - Buck, Thilo M.

AU - Vos, Rogier M.

AU - Alves, C. Henrique

AU - Wijnholds, Jan

N1 - Funding Information: The authors thank Ilse Voshart, Ariadna Rocha Sierra, Martijn Koomen, Charlotte A. Andriessen, and Annelies Boonzaier-van der Laan for technical assistance, and all Wijnholds Lab members for advice on the manuscript. This work was supported by the Foundation Fighting Blindness ( TA-GT-0715-0665-LUMC , to J.W.), the Netherlands Organisation for Health Research and Development (ZonMw grant 43200004 , to J.W.), and by the Dutch blindness funds ( Uitzicht 2013-13 , to J.W. and Uitzicht 2018-6 , to C.H.A. and J.W.): Rotterdamse Stichting Blindenbelangen , MaculaFonds , Stichting Blindenhulp , Landelijke Stichting voor Blinden en Slechtzienden , Algemene Nederlandse Vereniging ter Voorkoming van Blindheid , and Stichting Blinden-Penning . Publisher Copyright: © 2020 The Author(s) Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021

Y1 - 2021

N2 - Loss of Crumbs homolog 1 (CRB1) or CRB2 proteins in Müller cells or photoreceptors in the mouse retina results in a CRB dose-dependent retinal phenotype. In this study, we present a novel Müller cell-specific Crb1KOCrb2LowMGC retinitis pigmentosa mouse model (complete loss of CRB1 and reduced levels of CRB2 specifically in Müller cells). The Crb double mutant mice showed deficits in electroretinography, optokinetic head tracking, and retinal morphology. Exposure of retinas to low levels of DL-α-aminoadipate acid induced gliosis and retinal disorganization in Crb1KOCrb2LowMGC retinas but not in wild-type or Crb1-deficient retinas. Crb1KOCrb2LowMGC mice showed a substantial decrease in inner/outer photoreceptor segment length and optokinetic head-tracking response. Intravitreal application of rAAV vectors expressing human CRB2 (hCRB2) in Müller cells of Crb1KOCrb2LowMGC mice subsequently exposed to low levels of DL-α-aminoadipate acid prevented loss of vision, whereas recombinant adeno-associated viral (rAAV) vectors expressing human CRB1 (hCRB1) did not. Both rAAV vectors partially protected the morphology of the retina. The results suggest that hCRB expression in Müller cells is vital for control of retinal cell adhesion at the outer limiting membrane, and that the rAAV-cytomegalovirus (CMV)-hCRB2 vector is more potent than rAAV-minimal CMV (CMVmin)-hCRB1 in protection against loss of vision.

AB - Loss of Crumbs homolog 1 (CRB1) or CRB2 proteins in Müller cells or photoreceptors in the mouse retina results in a CRB dose-dependent retinal phenotype. In this study, we present a novel Müller cell-specific Crb1KOCrb2LowMGC retinitis pigmentosa mouse model (complete loss of CRB1 and reduced levels of CRB2 specifically in Müller cells). The Crb double mutant mice showed deficits in electroretinography, optokinetic head tracking, and retinal morphology. Exposure of retinas to low levels of DL-α-aminoadipate acid induced gliosis and retinal disorganization in Crb1KOCrb2LowMGC retinas but not in wild-type or Crb1-deficient retinas. Crb1KOCrb2LowMGC mice showed a substantial decrease in inner/outer photoreceptor segment length and optokinetic head-tracking response. Intravitreal application of rAAV vectors expressing human CRB2 (hCRB2) in Müller cells of Crb1KOCrb2LowMGC mice subsequently exposed to low levels of DL-α-aminoadipate acid prevented loss of vision, whereas recombinant adeno-associated viral (rAAV) vectors expressing human CRB1 (hCRB1) did not. Both rAAV vectors partially protected the morphology of the retina. The results suggest that hCRB expression in Müller cells is vital for control of retinal cell adhesion at the outer limiting membrane, and that the rAAV-cytomegalovirus (CMV)-hCRB2 vector is more potent than rAAV-minimal CMV (CMVmin)-hCRB1 in protection against loss of vision.

KW - AAV

KW - adeno-associated viral vector

KW - CRB1

KW - CRB2

KW - DL-AAA

KW - Müller glial cell

KW - retinitis pigmentosa

UR - http://www.scopus.com/inward/record.url?scp=85100022804&partnerID=8YFLogxK

U2 - 10.1016/j.omtm.2020.12.012

DO - 10.1016/j.omtm.2020.12.012

M3 - Article

AN - SCOPUS:85100022804

SN - 2329-0501

VL - 20

SP - 423

EP - 441

JO - Molecular Therapy - Methods and Clinical Development

JF - Molecular Therapy - Methods and Clinical Development

ER -