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Da Sun Thesis 03272018.pdf (8.88 MB)

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DEVELOPMENT OF GENE THERAPIES FOR INHERITED RETINAL DEGENERATIONS: A NON-VIRAL APPROACH

Sun, Da, Sun
http://rave.ohiolink.edu/etdc/view?acc_num=case1522331751836337

Abstract Details

2018, Doctor of Philosophy, Case Western Reserve University, Biomedical Engineering.
Inherited retinal degenerations (IRDs) are a major cause of blindness, which can be caused by mutations in a single gene occurred in rod/cone photoreceptor cells (PR) or retinal pigment epithelium (RPE). IRDs can be treated with gene replacement therapy (GRT) due to easy localized delivery into the retina and its immune privilege. Development of safe and effective gene GRT is critical in this field. In this work, we evaluated three non-viral GRTs for treatments of IRDs. First, a hybrid system of a multifunctional lipid ECO and a G4 nanoglobule was designed for efficient gene delivery into RPE cells at low charge ratios. This system formed stable nanoparticles at low N/P ratios, exhibited low cytotoxicity, and induced high GFP expression in ARPE-19 cells at N/P = 6. The hybrid nanoparticles mediated significant reporter gene GFP expression ex-vivo in the retina from wild type C57BL/6J mice and in vivo in BALB/c mice. Second, we developed a targeted non-viral GRT using a multifunctional lipid ECO for treating Leber’s congenital amaurosis type 2 (LCA2). ECO formed stable nanoparticles with plasmid DNA (pDNA) at a low N/P ratio and mediated high gene transfection efficiency in ARPE-19 cells. All-trans-retinylamine was incorporated into the nanoparticles for targeted delivery of pDNA into the RPE. The targeted ECO/pDNA nanoparticles provided high GFP expression in the RPE of Rpe65-/- mice after subretinal injection. GRT using targeted ECO system increased electroretinographic activity for at least 120 days. A safety study in wild-type BALB/c mice indicated no irreversible retinal damage following subretinal injection of these targeted nanoparticles. Third, we developed an ECO based GRT for Stargardt’s Disease (STGD). ECO could stably encapsulate large ABCA4 gene and induce efficient gene transfection. A rhodopsin promoter was incorporated into the plasmid to generate specific gene expression to the photoreceptors. Evaluations with reporter genes demonstrated enhanced and tissue specific expressions in vitro and in vivo. Subretinal treatment of ECO/pABCA4 GRT demonstrated high and photoreceptor specific expression of therapeutic ABCA4 gene in Abca4-/- mice, and prevented A2E accumulations at least for 6 months. Safety studies demonstrated excellent in vitro and in vivo safety profiles of ECO/pABCA4 based GRT.
Eben Alsberg, Dr (Committee Member)
Timothy Kern, Dr (Committee Member)
Akiko Maeda, Dr (Committee Member)
Stathis Karathanasis, Dr (Committee Chair)
Zheng-Rong Lu, Dr (Advisor)
205 p.
Biomedical Engineering

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Citations

  • Sun, Sun, D. (2018). DEVELOPMENT OF GENE THERAPIES FOR INHERITED RETINAL DEGENERATIONS: A NON-VIRAL APPROACH [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1522331751836337

    APA Style (7th edition)

  • Sun, Sun, Da. DEVELOPMENT OF GENE THERAPIES FOR INHERITED RETINAL DEGENERATIONS: A NON-VIRAL APPROACH. 2018. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1522331751836337.

    MLA Style (8th edition)

  • Sun, Sun, Da. "DEVELOPMENT OF GENE THERAPIES FOR INHERITED RETINAL DEGENERATIONS: A NON-VIRAL APPROACH." Doctoral dissertation, Case Western Reserve University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=case1522331751836337

    Chicago Manual of Style (17th edition)

case1522331751836337
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© 2018, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.