June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Pharmacological targeting of VCP in porcine and human organotypic retinal explants.
Author Affiliations & Notes
  • Ana Cristina Almansa Garcia
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
    Graduate Training Centre of Neuroscience, Tubingen, Baden-Württemberg, Germany
  • Bowen Cao
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
    Graduate Training Centre of Neuroscience, Tubingen, Baden-Württemberg, Germany
  • Merve Sen
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
  • Angela Armento
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
  • Sylvia Bolz
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
  • Blanca Arango Gonzalez
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
  • marius ueffing
    Molecular Biology of Retinal Degenerations, Universitatsklinikum Tubingen Forschungsinstitut fur Augenheilkunde, Tubingen, Baden-Württemberg, Germany
  • Footnotes
    Commercial Relationships   Ana Cristina Almansa Garcia None; Bowen Cao None; Merve Sen None; Angela Armento None; Sylvia Bolz None; Blanca Gonzalez None; marius ueffing None
  • Footnotes
    Support  This study was supported by the ProRetina Foundation, the Kerstan Foundation, the Maloch Stiftung, and the EU’s H2020 research and innovation programme under GA No 863203 (DynAMic).
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 1517. doi:
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      Ana Cristina Almansa Garcia, Bowen Cao, Merve Sen, Angela Armento, Sylvia Bolz, Blanca Arango Gonzalez, marius ueffing; Pharmacological targeting of VCP in porcine and human organotypic retinal explants.. Invest. Ophthalmol. Vis. Sci. 2023;64(8):1517.

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

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Abstract

Purpose : We have recently discovered that pharmacological inhibition of valosin-containing protein (VCP) prevents photoreceptor degeneration and preserves rod photoreceptor function in rodent models of retinitis pigmentosa. To move this pre-clinical proof of concept closer to clinical application, we investigated the effect of VCP inhibition in cone-rich organotypic cultures of porcine and human retinas to assess neuroretinal protection of cones from RD as well as drug-induced toxicity.

Methods : Retinal explants from the cone-rich visual streak of enucleated adult porcine eyes and human retinal explants of a human donor eye enucleated to remove a uveal melanoma, were cultivated in a medium-air interphase and maintained in serum-free conditions for 8 or 4 days. Two structurally unrelated VCP inhibitors, ML240 and NMS-873, or DMSO vehicle control, were added to the medium every two days. Retinal morphology, cell death, and cone-photoreceptor survival were assessed over 8 days of treatment, applying immunostaining for rhodopsin, cone opsins, and Iba-1 (as a marker for microglia activation), TUNEL staining, and outer nuclear layer cell row quantification.

Results : Enucleation is accompanied by axotomy as well as by decoupling from the physiological blood supply. In consequence, progressive retinal degeneration can be observed in retinal explants of porcine or human origin marked by an increase in TUNEL-positive cells, degeneration of rods and cones, and shortening of photoreceptor outer segments. VCP inhibition significantly reduced TUNEL-positive neuroretinal cells, indicating increased cell survival over time. Moreover, we could observe an increase in cone opsin expression, as well as significantly reduced activation of microglia cells. Likewise, VCP inhibition in human retinal explants resulted in a significantly higher number of photoreceptor nuclei concomitant with reduced TUNEL-positive cells. Specifically, cones were able to profit from this treatment displaying longer outer segments.

Conclusions : Pharmacological VCP inhibition protects rods and cones within porcine and human retinal organotypic cultures from retinal degeneration caused by enucleation and transfer to in vitro conditions. A long-lasting protection combined with a lack of any detectable cytotoxic side effects encourages us to further investigate the potential of VCP inhibition as a future therapeutical approach for human retinal disorders.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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