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rAAV-mediated over-expression of acid ceramidase prevents retinopathy in a mouse model of Farber lipogranulomatosis

Abstract

Farber disease (FD) is a rare monogenic lysosomal storage disorder caused by mutations in ASAH1 that results in a deficiency of acid ceramidase (ACDase) activity and the abnormal systemic accumulation of ceramide species, leading to multi-system organ failure involving neurological decline and retinopathy. Here we describe the effects of rAAV-mediated ASAH1 over-expression on the progression of retinopathy in a mouse model of FD (Asah1P361R/P361R) and its littermate controls (Asah1+/+ and Asah1+/P361R). Using a combination of non-invasive multimodal imaging, electrophysiology, post-mortem histology and mass spectrometry we demonstrate that ASAH1 over-expression significantly reduces central retinal thickening, ceramide accumulation, macrophage activation and limits fundus hyper-reflectivity and auto-fluorescence in FD mice, indicating rAAV-mediated over-expression of biologically active ACDase protein is able to rescue the anatomical retinal phenotype of Farber disease. Unexpectedly, ACDase over-expression in Asah1+/+ and Asah1+/P361R control eyes was observed to induce abnormal fundus hyper-reflectivity, auto-fluorescence and retinal thickening that closely resembles a FD phenotype. This study represents the first evidence of a gene therapy for Farber disease-related retinopathy. Importantly, the described gene therapy approach could be used to preserve vision in FD patients synergistically with broader enzyme replacement strategies aimed at preserving life.

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Fig. 1: ACDase expression and activity assay following in vivo or in vitro sc-rAAV2/2[MAX].ASAH1 treatment.
Fig. 2: cSLO imaging of sc-rAAV2/2[MAX].ASAH1 and PBS injected eyes.
Fig. 3: OCT imaging of sc-rAAV2/2[MAX].ASAH1 and PBS injected eyes.
Fig. 4: Histochemistry and immunofluorescence on paraffin sections of Asah1P361R/P361R (MUT) mouse eye at PW 8-9.
Fig. 5: ERG analysis of sc-rAAV2/2[MAX].ASAH1 and PBS injected eyes.
Fig. 6: Ceramide levels are reduced following sc-rAAV2/2[MAX].ASAH1 treatment.

Data availability

The authors declare that the data supporting the findings of this study are available within the article and its supplementary information files.

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Acknowledgements

The authors thank Lisa King, Christine Skumatz, Joseph Thulin, and the Biomedical Resource Center at MCW for their help with animal care, in addition to Christine Duris, Qiuhui Yang, and the Children’s Research Institute histology core of MCW for their contributions to histological studies. We thank Benyapa Khowpinitchai and Alexander Salmon for their help on OCT data measurement and analysis. We thank Xuntian Jiang and the Metabolomics Core at Washington University (St. Louis, MO) for assistance with sphingolipid quantification.

Funding

This study was supported through intramural funds available to DML and JAM.

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HZ and MSN designed, conducted the experiments, and wrote the manuscript; HL and XP performed OCT data analysis and statistics; JAM supervised the study and paper review; DML designed the experiments, conducted the mice injection, and wrote the manuscript.

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Correspondence to Daniel M. Lipinski.

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Zhang, H., Nagree, M.S., Liu, H. et al. rAAV-mediated over-expression of acid ceramidase prevents retinopathy in a mouse model of Farber lipogranulomatosis. Gene Ther (2022). https://doi.org/10.1038/s41434-022-00359-w

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