June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Anti-ceramide immunotherapy for diabetic retinopathy
Author Affiliations & Notes
  • Tim Florian Dorweiler
    Physiology, Michigan State University, East Lansing, Michigan, United States
  • Peter Nolan
    Physiology, Michigan State University, East Lansing, Michigan, United States
  • Richard N. Kolesnick
    Department of Molecular Pharmacology, Memorial Sloan Kettering Cancer Center, New York, New York, United States
  • Julia V Busik
    Physiology, Michigan State University, East Lansing, Michigan, United States
  • Footnotes
    Commercial Relationships   Tim Dorweiler None; Peter Nolan None; Richard Kolesnick Ceramedix, Code O (Owner); Julia Busik Ceramedix, Code C (Consultant/Contractor)
  • Footnotes
    Support  1R01EY030766
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 459. doi:
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      Tim Florian Dorweiler, Peter Nolan, Richard N. Kolesnick, Julia V Busik; Anti-ceramide immunotherapy for diabetic retinopathy. Invest. Ophthalmol. Vis. Sci. 2022;63(7):459.

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

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Abstract

Purpose : Previous work highlights the detrimental effects of increased ceramide generation in diabetic retinopathy (DR) progression. We have shown that ASM mediated ceramide generation leads to chronic low-grade inflammation in various cell types such as endothelial cells. We hypothesize that underlying mechanism is based on the formation of pro-inflammatory ceramide rich platforms (CRP). The focus of this project is to address the novel concept of functional and highly-specific ceramide inhibition using cutting edge anti-ceramide immunotherapy to prevent early onset diabetes induced pro-inflammatory and pro-apoptotic changes in the retinal vasculature.

Methods : Ischemia reperfusion (I/R) studies were utilized to mimic acute vascular degeneration in mice as observed in DR. Streptozotocin was utilized to induce hyperglycemia-induced vascular degeneration as a type-1 diabetes model. Anti-ceramide single chain variable fragment (scFv) treatment was administered via a 2 ug single dose into the vitreous. Pro-inflammatory markers (IL-1β, VEGF and ICAM-1) as well as ASM expression levels were analyzed via qRT-PCR. Ceramide species were analyzed via lipidomics (nESI-MS/MS). Vascular degeneration was captured using FITC-albumin technique and confocal microcopy and quantified using MetaMorph software.

Results : In vivo I/R studies in asm+/+ and asm-/- mice identified significant upregulation of ASM, C16 and C18 ceramide production, as well as IL-1β, VEGF and ICAM-1 gene expression in asm+/+ but not in asm-/- mice (n=6 per group; p<0.05). To determine feasibility of anti-ceramide immunotherapy, intravitreal injection of anti-ceramide scFv was first performed in the mouse I/R model. We found significant upregulation of IL-1β, VEGF and ICAM-1 gene expression in WT untreated retinas injured by I/R. These pro-inflammatory changes were inhibited by a single injection of anti-ceramide scFv 12 hours prior to I/R injury (n=6-8 per group; p<0.05). Impact of intravitreal injection of scFv was next tested in STZ-induced Wistar rat diabetes model. A single injection of scFv at the onset of hyperglycemia prevented diabetes-induced increase in retinal vascular permeability (n=3-10 per group; p=0.08).

Conclusions : These results suggest that anti-ceramide scFv immunotherapy may represent a novel approach to targeting deleterious ceramide-rich platform mediated pro-inflammatory and pro-apoptotic changes prevalent in the early stages of vasculopathy in the diabetic retina.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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