Journal article
Development of gene augmentation strategy for the treatment of MAK associated Retinitis Pigmentosa
The FASEB journal, Vol.33(S1), pp.802.20-802.20
04/2019
DOI: 10.1096/fasebj.2019.33.1_supplement.802.20
Abstract
Purpose
The gene male germ cell associated kinase (MAK) encodes a protein involved in regulation of photoreceptor cilia length. By combining next generation whole exome sequencing and induced pluripotent stem cell (iPSC) technology we found that an Alu repeat inserted in exon 9 of the MAK gene resulted in a loss of normal MAK transcript and development of human autosomal recessive retinitis pigmentosa (RP). Although a relatively rare cause of disease in the general population, the MAK variant is enriched in individuals of Ashkenazi Jewish descent. In this population, 1 in 55 individuals are carriers and 1/3 of all cases of recessive RP is caused by this gene. The purpose of this study was to determine if a viral gene augmentation strategy could be used to restore functional MAK protein and in turn would be a good candidate for treatment of early stage MAK associated RP.
Methods
To demonstrate the ability of the developed MAK gene transfer vector to restore normal transcript and protein, patient specific iPSC derived photoreceptor precursor cells were transduced and immunocytochemistry, rt‐PCR and western blot analysis were performed. To demonstrate transgene function, cilia length assays were performed using patient derived fibroblast cells in vitro and MAK knock out zebra fish in vivo. In addition, visual function testing was performed in the zebra fish. To demonstrate safety of cGMP generated clinical grade vector, local and systemic toxicity studies were performed via subretinal injection into wildtype rats.
Results
MAK mutant RP iPSC derived photoreceptor cells harboring the previously identified ALU insertion (Tucker et. al. 2011) were generated and transduced with viral vectors containing the retinal MAK transcript. At 1‐week post‐transduction normal retinal MAK transcript and protein could be detected via rt‐PCR and western blotting respectively. Using patient derived fibroblast cells and MAK knockdown zebra fish we demonstrate that over expression of retinal MAK transgene restored primary cilia length. In addition, the visual defect identified in MAK knockdown zebra fish was mitigated via treatment with retinal MAK transgene. Following subretinal delivery of clinical grade vector into wild type rats no evidence of local or systemic toxicity was detected at either 1 or 3 months' post‐treatment.
Conclusions
We have successfully developed a MAK gene replacement strategy that has been validated in human iPSC derived photoreceptor precursor cells in vitro and MAK knock down zebra fish and wildtype rats in vivo.
This is from the Experimental Biology 2019 Meeting. There is no full text article associated with this published in The FASEB Journal.
Details
- Title: Subtitle
- Development of gene augmentation strategy for the treatment of MAK associated Retinitis Pigmentosa
- Creators
- Katherine N Gibson‐Corley - University of IowaCathryn Cranston - University of IowaKristin Anfinson - University of IowaEmily Kaalberg - University of IowaIan Han - University of IowaDiane C Slusarski - University of IowaAdam Goeken - University of IowaThomas Businga - University of IowaHannah Brown - University of IowaRobert F Mullins - University of IowaEdwin M Stone - University of IowaBudd A Tucker - University of Iowa
- Resource Type
- Journal article
- Publication Details
- The FASEB journal, Vol.33(S1), pp.802.20-802.20
- Publisher
- The Federation of American Societies for Experimental Biology
- DOI
- 10.1096/fasebj.2019.33.1_supplement.802.20
- ISSN
- 0892-6638
- eISSN
- 1530-6860
- Number of pages
- 1
- Language
- English
- Date published
- 04/2019
- Academic Unit
- Ophthalmology and Visual Sciences; Iowa Neuroscience Institute; Biology; Pathology
- Record Identifier
- 9984071674002771
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