June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Towards non-invasive, magnetic-assisted delivery of mRNA to the retina by topical application
Author Affiliations & Notes
  • Marco Bassetto
    Physiology and Biophysics, University of California Irvine School of Medicine, Irvine, California, United States
    Research service, VA Long Beach Healthcare System, Long Beach, California, United States
  • Carolline Rodrigues Menezes
    Ophthalmology, University of California Irvine School of Medicine, Irvine, California, United States
  • Dominik Lewandowski
    Gavin Herbert Eye Institute, Irvine, California, United States
    Physiology and Biophysics, University of California Irvine School of Medicine, Irvine, California, United States
  • Leonor Anahi Leyva
    Physiology and Biophysics, University of California Irvine School of Medicine, Irvine, California, United States
  • Jianying Kiser
    Ophthalmology, University of California Irvine School of Medicine, Irvine, California, United States
  • Philip David Kiser
    Physiology and Biophysics, University of California Irvine School of Medicine, Irvine, California, United States
    Research service, VA Long Beach Healthcare System, Long Beach, California, United States
  • Footnotes
    Commercial Relationships   Marco Bassetto None; Carolline Rodrigues Menezes None; Dominik Lewandowski None; Leonor Leyva None; Jianying Kiser None; Philip Kiser None
  • Footnotes
    Support  Knights Templar Eye Foundation - Pediatric ophthalmology career-starter research grant - 2022
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2615. doi:
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    • Get Citation

      Marco Bassetto, Carolline Rodrigues Menezes, Dominik Lewandowski, Leonor Anahi Leyva, Jianying Kiser, Philip David Kiser; Towards non-invasive, magnetic-assisted delivery of mRNA to the retina by topical application. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2615.

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

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Abstract

Purpose : Eyedrops are non-invasive but inefficient to target the retina. Previous data showed that magnetic-assisted delivery of hydrophilic small drug molecules formulated with magnetic nanoparticles (MNPs) improves the efficacy of eyedrops to treat retinal degeneration in mice. Here we explored the universality of this method to deliver messenger RNA (mRNA) to the retina of mice after topical application.

Methods : Cre mRNA was used as model for mRNA delivery. MNPs were synthesized by co-precipitation of Fe(II) and Fe(III) in alkaline medium. The nanocrystals were coated with proprietary cationic polymers or lipids to generate four formulations. MNP's hydrodynamic size and surface electrostatic charge were analyzed by dynamic light scattering (DLS) and electrophoretic DLS. Two retinal cell lines, 661W and ARPE19, expressing a Cre-responsive LoxP-GFP-stop-LoxP-RFP-stop reporter system were generated to screen the MNPs for efficient Cre mRNA transfection in vitro. Lipid-coated MNPs (1 µg mRNA/0.1µL MNPs in 10 µL) were applied topically under the influence of a magnetic field (<1T) for 30 min on B6.129 mT/mG mice. The magnet was placed at the base of the skull. 1µL of the same formulation was injected in the subretinal space as a positive control. Transfection efficiency was assessed 7 weeks post treatment by fluorescence microscopy. Ocular structures were assessed by optical coherence tomography (OCT) and scanning laser ophthalmoscopy (SLO).

Results : The MNPs were 150 ± 30 nm in size and all had a positive charge of 50 ± 15 mV. Lipid-based MNPs were the most efficient to deliver Cre mRNA in both 661W and ARPE19 cells at the ratio 0.1 µg mRNA/0.1 µg MNP. This ratio induced color switching in 72 ± 15 % and 82 ± 11 % of cells respectively with negligible toxicity. Subretinal injection and topical application of this formulation did not cause ocular abnormalities by OCT, SLO and histology after 1 week. Transfection was detected at the level of retina pigment epithelium (RPE) and photoreceptors after subretinal injection. Topical application failed to induce color switching in the retina.

Conclusions : Lipid-based MNPs allow efficient mRNA transfection in vitro and in vivo. Topical application of higher mRNA and MNPs concentrations should be tested. Delivery of Cre protein may enhance the transfection efficiency in the retina in virtue of its significantly lower molecular weight compared to mRNA.

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

 

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