Delivering topical IL-33 with a cell penetrating peptide to treat neovascular age-related macular degeneration.
Thomas, Chloe Weston, Christopher Cooklin, Jessica Copland, Dave Dick, Andrew Denniston, Alastair Hill, Lisa
Thomas, Chloe
Weston, Christopher
Cooklin, Jessica
Copland, Dave
Dick, Andrew
Denniston, Alastair
Hill, Lisa
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Abstract
Abstract
Purpose : Age-related macular degeneration (AMD) is the most common cause of central vision loss globally. Neovascular AMD (nAMD) is currently treated with regular intraocular injections of anti-VEGF therapies, to which some patients are unresponsive, while multiple injections are not well-tolerated and can cause complications including infection. We previously demonstrated intraocular delivery of recombinant interleukin-33 (IL-33) reduced laser-induced choroidal neovascularisation (CNV) pathology in a mouse model, offering a promising new treatment. Additionally, an eyedrop of bevacizumab complexed to a cell penetrating peptide (CPP) reduced lesion size equivalent to intravitreal injection delivery in a mouse CNV model. Thus, we propose combining IL-33 immunotherapy and our delivery agent, IL-33+CPP6, to be developed as a topical formulation.
Methods : Human recombinant IL-33 was complexed with the CPP to assess cell toxicity, MTT and LDH assays for 48h over a dose range of 0.2–200pg/ul in cultured human RPE (ARPE-19) and human ocular choroidal fibroblasts (HOCFs). HOCF scratch assay was performed and cells treated with media, vehicle, CPP, IL-33 (200pg/ul) or IL-33+CPP and imaged over 48h. The percentage wound closure (end vs start width) was measured using Cell IQ software. N=3, repeated on 3 independent occasions.
Results : IL-33+CPP complexes were non-toxic to ARPE-19 or HOCFs across a dose range (0.2–200pg/ul). In ARPE-19 cells and HOCFs, there were no differences after treatment in total viable cell number (total cell LDH; P=0.8306, P=0.9641, ANOVA) or significant changes in cellular viability (MTT assay; P=0.9772, P=0.2910, ANOVA). Importantly IL-33+CPP had equivalent significant biological effects to IL-33 alone, reducing wound healing (~15% less wound closure at 48h vs media/vehicle/CPP6). Across groups in scratch assay, there was a significant effect on percentage wound closure (P=0.0430, ANOVA), with borderline significance in post-hoc analysis (Tukey) between media and IL-33 (P=0.0529) and IL-33+CPP (P=0.0639) treatments.
Conclusions : IL-33+CPP is a promising treatment in a formulation of a drop for nAMD. IL-33+CPP demonstrates non-toxicity and has preserved biological effect as direct IL-33 administration. Further experiments will assess ocular penetration and in vivo efficacy of our topical administration.
Citation
Thomas, C. N., Theodoropoulou, S., Weston, C., Cooklin, J., Copland, D., Dick, A. D., Denniston, A. K., & Hill, L. J. (2021). Delivering topical IL‑33 with a cell‑penetrating peptide to treat neovascular age‑related macular degeneration. Investigative Ophthalmology & Visual Science, 62(8), Article 212.