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Intravitreal Delivery of PEGylated-ECO Plasmid DNA Nanoparticles for Gene Therapy of Stargardt Disease

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Abstract

Objective

Current gene therapy of inherited retinal diseases is achieved mainly by subretinal injection, which is invasive with severe adverse effects. Intravitreal injection is a minimally invasive alternative for gene therapy of inherited retinal diseases. This work explores the efficacy of intravitreal delivery of PEGylated ECO (a multifunctional pH-sensitive amphiphilic amino lipid) plasmid DNA (pGRK1-ABCA4-S/MAR) nanoparticles (PEG-ELNP) for gene therapy of Stargardt disease.

Methods

Pigmented Abca4−/− knockout mice received 1 µL of PEG-ELNP solution (200 ng/uL, pDNA concentration) by intravitreal injections at an interval of 1.5 months. The expression of ABCA4 in the retina was determined by RT-PCR and immunohistochemistry at 6 months after the second injection. A2E levels in the treated eyes and untreated controls were determined by HPLC. The safety of treatment was monitored by scanning laser ophthalmoscopy and electroretinogram (ERG).

Results

PEG-ELNP resulted in significant ABCA4 expression at both mRNA level and protein level at]6 months after 2 intravitreal injections, and a 40% A2E accumulation reduction compared with non-treated controls. The PEG-ELNP also demonstrated excellent safety as shown by scanning laser ophthalmoscopy, and the eye function evaluation from electroretinogram.

Conclusions

Intravitreal delivery of the PEG-ELNP of pGRK1-ABCA4-S/MAR is a promising approach for gene therapy of Stargardt Disease, which can also be a delivery platform for gene therapy of other inherited retinal diseases.

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Data Availability

The datasets analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This project was supported by the Gund-Harrington Scholars Award from the Harrington Discovery Institute and the Foundation Fighting Blindness, and National Cancer Institute R01CA235152. ZRL is an M. Frank Rudy and Margaret Domiter Rudy Professor of Biomedical Engineering.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, 44106, United States

    Da Sun, Wenyu Sun, Song-Qi Gao, Jonathan Lehrer, Hong Wang, Ryan Hall & Zheng-Rong Lu

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  1. Da Sun
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  2. Wenyu Sun
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Contributions

Z.R.L., D.S. and W.S. conceived of the strategy, and designed the experiments. D.S and W.S. were involved in all aspects of this work. J.L. performed analysis of nanoparticle formulations. W.S., D.S. S.G. provided help on ERG examinations. D.S and C.N. performed SLO. R.H. and H.W. provided help on PCR and data analysis. All authors read and approved the final manuscript.

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Correspondence to Zheng-Rong Lu.

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Conflict of Interest

The gene therapy reported in this work was licensed to Helios BioPharmaceuticals for commercialization. ZRL may have ownership interest in the company.

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Sun, D., Sun, W., Gao, SQ. et al. Intravitreal Delivery of PEGylated-ECO Plasmid DNA Nanoparticles for Gene Therapy of Stargardt Disease. Pharm Res 41, 807–817 (2024). https://doi.org/10.1007/s11095-024-03679-1

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