Abstract
Purpose :
The therapeutic targets of age-related macular degeneration (AMD) remain to be investigated, considering the potential side effect and resistance of recent therapy. Transfer RNA derived small non-coding RNAs (tsRNAs) is a novel cluster of non-coding RNAs which regulate angiogenesis and cell aging. We identify a tsRNA, tRF-Glu-CTC, in the development of choroidal neovasculation (CNV) and hypothesize that the inhibition of it can alleviate neovascular AMD by improving the functional amelioration of choroidal vessel.
Methods :
C57BL/6J mice (6 weeks age, male) were used to build laser-induced CNV model. tRF-Glu-CTC agomir or antagomir were intravitreally injected in eyes of experimental group (N=5-8) to regulate the level of tRF-Glu-CTC. Isolectin B4 (IB4) was used to stain the area of neovasculation in chorod flat-mounts. Primary HUVECs were used to detected migration and angigenesis of endothelial cells via Transwell and tube-forming assay (N=4). Transcriptome sequencing was performed to screen out the target of tRF-Glu-CTC. Two-tailed Student’s t-test or one-way ANOVA test were used for statistical analysis.
Results :
A dramatic increase of CNV area (23732±1494 μm2, P = 0.012) was found in mice injected with tRF-Glu-CTC agomir, while antagomir contributed to a 44 % decrease of CNV (7555±840.8 μm2, P=0.04) compared to control groups (13380±1661 μm2), which showed similar anti-angiogenic effect as bevacizumab (8819±1046 μm2). tRF-Glu-CTC mimic increased the number of migrated cells (431.3±55.17, 224±8.5 for control group, P = 0.0024) and the total branches length in tube-forming assay (12300±143.6, 11081±301.2 for control group, P = 0.01) in HUVECs compared to control. Sequencing analysis revealed that tRF-Glu-CTC was positive correlated with the increase of inflammatory senescence-associated secretory phenotype, including IL1β (6.63±1.14 fold change, P = 0.0081), IL6 (2.12±0.13 fold change, P = 0.0035), ICAM1 (1.88±0.26 fold change, P = 0.033).
Conclusions :
The upregulation of tRF-Glu-CTC aggravates the endothelial dysfunction by inducing the secretion of inflammatory factors and promoting the anigogenesis of endothelial cells. The pathological neovasculation are significantly alleviated by the inhibition of tRF-Glu-CTC. Thus, tRF-Glu-CTC can be a potential target for the treatment of neovascular AMD.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.