A drug-tunable Flt23k gene therapy for controlled intervention in retinal neovascularization
| Autor: | Jinying, Chen, Fan-Li, Lin, Jacqueline Y K, Leung, Leilei, Tu, Jiang-Hui, Wang, Yu-Fan, Chuang, Fan, Li, Hsin-Hui, Shen, Gregory J, Dusting, Vickie H Y, Wong, Leszek, Lisowski, Alex W, Hewitt, Bang V, Bui, Jingxiang, Zhong, Guei-Sheung, Liu |
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| Rok vydání: | 2020 |
| Předmět: |
Vascular Endothelial Growth Factor A
Gene Transfer Techniques Genetic Therapy Dependovirus Retinal Neovascularization Cell Hypoxia Rats Sprague-Dawley Disease Models Animal Tetrahydrofolate Dehydrogenase HEK293 Cells Receptors Vascular Endothelial Growth Factor Protein Domains Intravitreal Injections Animals Humans Female Transgenes |
| Zdroj: | Angiogenesis. 24(1) |
| ISSN: | 1573-7209 |
| Popis: | Gene therapies that chronically suppress vascular endothelial growth factor (VEGF) represent a new approach for managing retinal vascular leakage and neovascularization. However, constitutive suppression of VEGF in the eye may have deleterious side effects. Here, we developed a novel strategy to introduce Flt23k, a decoy receptor that binds intracellular VEGF, fused to the destabilizing domain (DD) of Escherichia coli dihydrofolate reductase (DHFR) into the retina. The expressed DHFR(DD)-Flt23k fusion protein is degraded unless "switched on" by administering a stabilizer; in this case, the antibiotic trimethoprim (TMP). Cells transfected with the DHFR(DD)-Flt23k construct expressed the fusion protein at levels correlated with the TMP dose. Stabilization of the DHFR(DD)-Flt23k fusion protein by TMP was able to inhibit intracellular VEGF in hypoxic cells. Intravitreal injection of self-complementary adeno-associated viral vector (scAAV)-DHFR(DD)-Flt23k and subsequent administration of TMP resulted in tunable suppression of ischemia-induced retinal neovascularization in a rat model of oxygen-induced retinopathy (OIR). Hence, our study suggests a promising novel approach for the treatment of retinal neovascularization. Schematic diagram of the tunable system utilizing the DHFR(DD)-Flt23k approach to reduce VEGF secretion. a The schematic shows normal VEGF secretion. b Without the ligand TMP, the DHFR(DD)-Flt23k protein is destabilized and degraded by the proteasome. c In the presence of the ligand TMP, DHFR(DD)-Flt23k is stabilized and sequestered in the ER, thereby conditionally inhibiting VEGF. Green lines indicate the intracellular and extracellular distributions of VEGF. Blue lines indicate proteasomal degradation of the DHFR(DD)-Flt23k protein. Orange lines indicate the uptake of cell-permeable TMP. TMP, trimethoprim; VEGF, vascular endothelial growth factor; ER, endoplasmic reticulum. |
| Databáze: | OpenAIRE |
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