scholarly journals Defining Phenotype, Tropism, and Retinal Gene Therapy Using Adeno-Associated Viral Vectors (AAVs) in New-Born Brown Norway Rats with a Spontaneous Mutation in Crb1

2021 ◽  
Vol 22 (7) ◽  
pp. 3563
Author(s):  
Nanda Boon ◽  
C. Henrique Alves ◽  
Aat A. Mulder ◽  
Charlotte A. Andriessen ◽  
Thilo M. Buck ◽  
...  

Mutations in the Crumbs homologue 1 (CRB1) gene cause inherited retinal dystrophies, such as early-onset retinitis pigmentosa and Leber congenital amaurosis. A Brown Norway rat strain was reported with a spontaneous insertion-deletion (indel) mutation in exon 6 of Crb1. It has been reported that these Crb1 mutant rats show vascular abnormalities associated with retinal telangiectasia and possess an early-onset retinal degenerative phenotype with outer limiting membrane breaks and focal loss of retinal lamination at 2 months of age. Here, we further characterized the morphological phenotype of new-born and adult Crb1 mutant rats in comparison with age-matched Brown Norway rats without a mutation in Crb1. A significantly decreased retinal function and visual acuity was observed in Crb1 mutant rats at 1 and 3 months of age, respectively. Moreover, in control rats, the subcellular localization of canonical CRB1 was observed at the subapical region in Müller glial cells while CRB2 was observed at the subapical region in both photoreceptors and Müller glial cells by immuno-electron microscopy. CRB1 localization was lost in the Crb1 mutant rats, whereas CRB2 was still observed. In addition, we determined the tropism of subretinal or intravitreally administered AAV5-, AAV9- or AAV6-variant ShH10Y445F vectors in new-born control and Crb1 mutant rat retinas. We showed that subretinal injection of AAV5 and AAV9 at postnatal days 5 (P5) or 8 (P8) predominantly infected the retinal pigment epithelium (RPE) and photoreceptor cells; while intravitreal injection of ShH10Y445F at P5 or P8 resulted in efficient infection of mainly Müller glial cells. Using knowledge of the subcellular localization of CRB1 and the ability of ShH10Y445F to infect Müller glial cells, canonical hCRB1 and hCRB2 AAV-mediated gene therapy were explored in new-born Crb1 mutant rats. Enhanced retinal function after gene therapy delivery in the Crb1 rat was not observed. No timely rescue of the retinal phenotype was observed using retinal function and visual acuity, suggesting the need for earlier onset of expression of recombinant hCRB proteins in Müller glial cells to rescue the severe retinal phenotype in Crb1 mutant rats.

2015 ◽  
Vol 24 (11) ◽  
pp. 3104-3118 ◽  
Author(s):  
Lucie P. Pellissier ◽  
Peter M. Quinn ◽  
C. Henrique Alves ◽  
Rogier M. Vos ◽  
Jan Klooster ◽  
...  

2008 ◽  
Vol 9 (3) ◽  
Author(s):  
Norbert Fülöp ◽  
Wenguang Feng ◽  
Dongqi Xing ◽  
Kai He ◽  
László G. Nőt ◽  
...  

2014 ◽  
Vol 135 ◽  
pp. 180-188 ◽  
Author(s):  
Robert L. Thunhorst ◽  
Terry Beltz ◽  
Alan Kim Johnson

1994 ◽  
Vol 93 (2) ◽  
pp. 464-469 ◽  
Author(s):  
Hideo Tsukagoshi ◽  
Tatsuo Sakamoto ◽  
Wenbing Xu ◽  
Peter J. Barnes ◽  
K.Fan Chung

1993 ◽  
Vol 6 (4) ◽  
pp. 201-213 ◽  
Author(s):  
A. Reichenbach ◽  
J.-U. Stolzenburg ◽  
W. Eberhardt ◽  
T.I. Chao ◽  
D. Dettmer ◽  
...  

2001 ◽  
Vol 8 (4) ◽  
pp. 762-767 ◽  
Author(s):  
R. de Waard ◽  
J. Garssen ◽  
J. Snel ◽  
G. C. A. M. Bokken ◽  
T. Sako ◽  
...  

ABSTRACT In this study, the effects of orally administered viableLactobacillus casei Shirota strain YIT9029 on the immunity parameters of Wistar and Brown Norway rats were examined. For this purpose, we used the Trichinella spiralis host resistance model. Two weeks before and during T. spiralisinfection, rats were fed 109 viable L. casei bacteria 5 days per week. The T. spiralis-specific delayed-type hypersensitivity (DTH) response was significantly enhanced in both Wistar and Brown Norway rats given L. casei. In both rat strains fedL. casei, serum T. spiralis-specific immunoglobulin G2b (IgG2b) concentrations were also significantly increased. In the model, no significant effects ofL. casei on larval counts or inflammatory reactions in the tongue musculature, body weights, or lymphoid organ weights were observed. Serum specific antibody responses, other than IgG2b, were not changed by feeding of L. casei. In contrast toL. casei, it was shown that orally administeredBifidobacterium breve or Bifidobacterium bifidum had no influence on the measured infection and immunity indices in the rat infection model. Since the rat DTH response is considered to be a manifestation of Th1 cell-mediated immunity and the IgG2b isotype has been associated with Th1 activity, it was concluded that Th1 cells could play an active role in the immunomodulatory effects of orally administered L. casei. Furthermore, our data do not indicate that the effect of oral supplementation withL. casei is dependent on the genetic background of the host.


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