An ES-LV-EPC Gene Therapy Agent for Suppression of Neovascularization in Oxygen-Induced Rat Models

Transplantation of gene transfected endothelial progenitor cells (EPCs) has provided a novel method for neovascularization (NV) therapy but not for ocular diseases. This study investigated the efficacy of endostatin (ES) transfected EPCs in retinal NV therapy. EPCs were transfected with ES-lentivirus (LV) recombinant construct to obtain ES-LV-EPCs. Oxygen-induced retinopathy rats received the same volume of intravitreal injections of empty-LV, ES-LV, EPCs, and ES-LV-EPCs. Retinal NV was compared using fundus fluoresce in angiography and hematoxylin-eosin staining method. Retinal expression of ES, vascular endothelial growth factor (VEGF), and CD31 were evaluated by immunohistochemistry method. QRT-PCR showed a high expression of ES in ES-LV-EPCs. The NV leakage area and the number of pre-retinal neovascular cell nuclei significantly decreased in ES-LV and ES-LV-EPCs groups, and the effects of ES-LV and ES-LV-EPCs on inhibiting retinal NV were almost the same. ES-LV group and ES-LV-EPCs group showed much more retinal distribution of ES. Intravitreal injections of ES-LV-EPCs inhibited retinal NV, VEGF or CD31 expression, and increased ES expression in vivo. ES-LV-EPC targeted and prevented pathologic retinal NV, and exerted a much safer gene therapy by using lentivirus transfected EPCs. Thus, gene combined EPCs represent a potential new therapeutic agent for the treatment of neovascular eye diseases. Funding Statement: This work was supported by National Natural Science Foundation of China (81570822, 81571819); Zhejiang Key Laboratory Fund of China (2011E10006); Zhejiang Province Key Research and Development Program (2015C03042); Zhejiang Provincial Natural Science Foundation of China (LZ18H180001, LY16H120002); National S&T Major Project of China ( NO.SQ2018ZX100301) Declaration of Interests: The authors declare that there are no conflicts of interest. Ethics Approval Statement: All procedures were performed in accordance with the ARVO statement for the use of Animals in Ophthalmic and Vision Research. The Institutional Animal Care and Ethics Committee of Zhejiang University (Hangzhou, China) approved all animal experiments.