June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa.
Author Affiliations & Notes
  • Budd A. Tucker
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Erin R Burnight
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Cathryn M Cranston
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Jessica L Fick
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Austin J Reutzel
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Melette R. Devore
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Trudi Westfall
    Biology, University of Iowa, Iowa City, Iowa, United States
  • C. Anthony Scott
    Biology, University of Iowa, Iowa City, Iowa, United States
  • Autumn Marsden
    Biology, University of Iowa, Iowa City, Iowa, United States
  • Katherine N Gibson-Corley
    Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Luke A Wiley
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Ian Han
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Diane Slusarski
    Biology, University of Iowa, Iowa City, Iowa, United States
  • Robert F Mullins
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Edwin M Stone
    Institute for Vision Research/Ophthalmology and Visual Science, The University of Iowa, Iowa City, Iowa, United States
  • Footnotes
    Commercial Relationships   Budd Tucker, None; Erin Burnight, None; Cathryn Cranston, None; Jessica Fick, None; Austin Reutzel, None; Melette Devore, None; Trudi Westfall, None; C. Anthony Scott, None; Autumn Marsden, None; Katherine Gibson-Corley, None; Luke Wiley, None; Ian Han, None; Diane Slusarski, None; Robert Mullins, None; Edwin Stone, None
  • Footnotes
    Support  Ruby Chair in Regenerative Ophthalmology
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 1485. doi:
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    • Get Citation

      Budd A. Tucker, Erin R Burnight, Cathryn M Cranston, Jessica L Fick, Austin J Reutzel, Melette R. Devore, Trudi Westfall, C. Anthony Scott, Autumn Marsden, Katherine N Gibson-Corley, Luke A Wiley, Ian Han, Diane Slusarski, Robert F Mullins, Edwin M Stone; Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa.. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1485.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : 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 results in a loss of normal MAK transcript and development of human autosomal recessive retinitis pigmentosa (RP). The purpose of this study was to determine if a viral gene augmentation strategy could be used to safely restore functional MAK protein as a step toward a treatment for early stage MAK-associated RP.

Methods : Patient-specific iPSC-derived photoreceptor precursor cells were transduced with the MAK gene transfer vector, and immunocytochemistry, rt-PCR and western blot analysis were performed to evaluate the effect on MAK expression. To demonstrate transgene function, cilia length assays were performed using patient-derived fibroblast cells in vitro and mak knockdown zebrafish in vivo. In addition, visual function testing was also performed in the zebrafish. Local and systemic toxicity studies of cGMP clinical grade MAK vector were performed via subretinal injection into wildtype rats.

Results : MAK mutant 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. One week after 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 the retinal MAK transgene restored primary cilia length in both MAK RP patient-derived fibroblasts and mak knockdown zebrafish. In addition, the visual defect in mak knockdown zebrafish was mitigated via treatment with the retinal MAK transgene. There was no evidence of local or systemic toxicity at 1-month or 3-months following subretinal delivery of clinical grade vector into wild type rats.

Conclusions : We developed a MAK gene replacement strategy and validated it in human iPSC-derived photoreceptor precursor cells in vitro as well as mak knockdown zebra fish and wildtype rats in vivo. The findings reported here provide the efficacy and safety data required for initiation of a phase 1 clinical trial for the treatment of patients with MAK-associated RP.

This is a 2021 ARVO Annual Meeting abstract.

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