Analysis of retinal expression of genes involved in the regulation of angiogenesis and endothelial barrier function following exposure of the retina to 577 nm wavelength laser light in continuous mode

Objective. The aim of the study was to investigate the retinal expression of genes involved in the regulation of angiogenesis and endothelial barrier function after exposure of the retina to 577 nm laser light in a continuous mode in experimental conditions after intravitreal injection of VEGF165 (Vascular endothelial growth factor).

Materials and methods. The study was performed on 4-5 week old male C57BL / 6J mice. 4 groups, 5 mice in each, were formed, one animal eye was experimental and the second remained intact. The animals of the first group received intravitreal injection of phosphate-buffered saline (PBS); animals in the second, third and fourth groups received intravitreal injection of 50 ng/ml of recombinant VEGF165; in the third and fourth groups, one day after the VEGF165 intravitreal administration, laser irradiation with a wavelength of 577 nm was performed on the retina in micropulse and continuous modes, respectively. Tissue samples from animals of the first and second groups were taken 2 days after PBS and VEGF165 administration, and in animals from the third and fourth groups one day after laser exposure to the retina.

Results. Retinal expression of genes involved in the regulation of angiogenesis and endothelial barrier function as a result of retinal exposure to 577 nm laser radiation in continuous mode following intravitreal injection of VEGF165 was studied. Genes with significant changes in expression levels were identified (genes that regulate the processes of angiogenesis).

Conclusion. Understanding the mechanisms of modulation of retinal gene expression may help to identify the key regulatory factors providing therapeutic effects of laser radiation in continuous and micropulsed modes and provide the basis for future therapeutic tactics for retinal diseases.

1. Кузник Б.И., Хавинсон В.Х., Тарновская С.И., Линькова Н.С., Козина Л.С., Дьяконов М.М. Адгезивная молекула JAM-A и молекулярные механизмы возрастной патологии: обзор литературы и собственных данных. Успехи геронтологии. 2015;28(4):656-668

2. Binz N., Graham C.E., Simpson K., Lai Y.K., Shen W.Y., Lai C.M., Speed T.P., Rakoczy P.E. Long-term effect of therapeutic laser photocoagulation on gene expression in the eye. FASEB J. 2006;20(2):383-385. DOI: 10.1096/fj.05-3890fje

3. Boeckel J.N., Guarani V., Koyanagi M., Roexe T., Lengeling A., Schermuly R.T., Gellert P., Braun T. et al. Jumonji domain-containing protein 6 (Jmjd6) is required for angiogenic sprouting and regulates splicing of VEGF-receptor 1. Proc Natl Acad Sci U S A. 2011;108(8):3276-3281. DOI: 10.1073/pnas.1008098108

4. Boström K., Zebboudj A.F., Yao Y., Lin T.S., Tor-res A. Matrix GLA protein stimulates VEGF expression through increased transforming growth factor-beta1 activity in endothelial cells. J Biol Chem. 2004;279(51):52904-52913. DOI: 10.1074/jbc.M406868200

5. Cao R., Bråkenhielm E., Pawliuk R., Wariaro D., Post M.J., Wahlberg E., Leboulch P., Cao Y. Angiogenic synergism, vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2. Nat Med. 2003;9(5):604-613. DOI: 10.1038/nm848

6. Contois L.W., Nugent D.P., Caron J.M., Cretu A., Tweedie E., Akalu A., Liebes L., Friesel R. et al. Insulin-like growth factor binding protein-4 differentially inhibits growth factor-induced angiogenesis. J Biol Chem. 2012;287(3):1779-1789. DOI: 10.1074/jbc.M111.267732

7. Cooke V.G., Naik M.U., Naik U.P. Fibroblast growth factor-2 failed to induce angiogenesis in junctional adhesion molecule-A-deficient mice. Arterioscler Thromb Vasc Biol. 2006;26(9):2005-2011. DOI: 10.1161/01.ATV.0000234923.79173.99

8. Daniele L.L., Adams R.H., Durante D.E., Pugh E.N. Jr., Philp N.J. Novel distribution of junctional adhesion molecule-C in the neural retina and retinal pigment epithelium. J Comp Neurol. 2007;505(2):166-176. DOI: 10.1002/cne.21489

9. Díaz-Coránguez M., Liu X., Antonetti D.A. Tight Junctions in Cell Proliferation. Int J Mol Sci. 2019;20(23):5972. DOI: 10.3390/ijms20235972.

10. Ding X., Bai Y., Zhu X., Li T., Jin E., Huang L., Yu W., Zhao M. The effects of pleiotrophin in proliferative vitreoretinopathy. Graefes Arch Clin Exp Ophthalmol. 2017;255(5):873-884. DOI: 10.1007/s00417-016-3582-9

11. Early Treatment Diabetic Retinopathy Study Research Group Photocoagulation for diabetic macular edema. Early treatment diabetic retinopathy study report number 1. Arch Ophthalmol. 1985;(103):1796-1806.

12. Fu Y., Tang M., Xiang X., Liu K., Xu X. Glucose affects cell viability, migration, angiogenesis and cellular adhesion of human retinal capillary endothelial cells via SPARC. Exp Ther Med. 2019;17(1): 273-283. DOI: 10.3892/etm.2018.6970

13. Gallego B.I., Salazar J.J., de Hoz R., Rojas B., Ramírez A.I., Salinas-Navarro M., Ortín-Martínez A., Valiente-Soriano F.J. et al. IOP induces upregulation of GFAP and MHC-II and microglia reactivity in mice retina contralateral to experimental glaucoma. J Neuroinflammation. 2012;(9):92. DOI: 10.1186/1742-2094-9-92

14. Gavrilova N.A., Borzenok S.A., Zaletaev D.V., Solomin V.A., Gadzhieva N.S., Tishchenko O.E., Komova O.U., Zinov'eva A.V. Molecular genetic mechanisms of influence of laser radiation with 577 nm wavelength in a microimpulse mode on the condition of the retina. Exp Eye Res. 2019;(185):107650. DOI: 10.1016/j.exer.2019.04.018

15. Haase M., Fitze G. HSP90AB1: Helping the good and the bad. Gene. 2016;575(2 Pt 1):171-186. DOI: 10.1016/j.gene.2015.08.063

16. Hammes H.P., Du X., Edelstein D., Taguchi T., Matsumura T., Ju Q., Lin J., Bierhaus A. et al. Benfotiamine blocks three major pathways of hyperglycemic damage and prevents experimental diabetic retinopathy. Nat Med. 2003;9(3):294-299. DOI: 10.1038/nm834

17. Hedhli N., Falcone D.J., Huang B., Cesarman-Maus G., Kraemer R., Zhai H., Tsirka SE., Santambrogio L. et al. The annexin A2/S100A10 system in health and disease: emerging paradigms. J Biomed Biotechnol. 2012;(2012):406273. DOI: 10.1155/2012/406273

18. Kim C.W., Son K.N., Choi S.Y., Kim J. Human lactoferrin upregulates expression of KDR/Flk-1 and stimulates VEGF-A-mediated endothelial cell proliferation and migration. FEBS Lett. 2006;580(18):4332-4336. DOI: 10.1016/j.febslet.2006.06.091

19. Kozak I., Luttrull J.K. Modern retinal laser therapy. Saudi J Ophthalmol. 2015;29(2):137-146. DOI: 10.1016/j.sjopt.2014.09.001

20. Lennikov M.S., Lennikov A., Tang S., Huang H. Proteomics reveals ablation of placental growth factor inhibits the insulin resistance pathways in diabetic mouse retina. bioRxiv. 2018. DOI: 10.1101/338368.

21. Li D, Xie K., Zhang L., Yao X., Li H., Xu Q., Wang X., Jiang J. et al. Dual blockade of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2) exhibits potent anti-angiogenic effects. Cancer Lett. 2016;377(2):164-173. DOI: 10.1016/j.canlet.2016.04.036

22. Malecaze F., Mascarelli F., Bugra K., Fuhrmann G., Courtois Y., Hicks D. Fibroblast growth factor receptor deficiency in dystrophic retinal pigmented epithelium. J Cell Physiol. 1993;154(3):631-642. DOI: 10.1002/jcp.1041540323

23. Mandell K.J., Berglin L., Severson E.A., Edel-hauser H.F., Parkos C.A. Expression of JAM-A in the human corneal endothelium and retinal pigment epithelium: localization and evidence for role in barrier function. Invest Ophthalmol Vis Sci. 2007;48(9):3928-3936. DOI: 10.1167/iovs.06-1536

24. Miljkovic-Licina M., Hammel P., Garrido-Urbani S., Lee B.P., Meguenani M., Chaabane C., Bochaton-Piallat M.L., Imhof B.A. Targeting olfactomedin-like 3 inhibits tumor growth by impairing angiogenesis and pericyte coverage. Mol Cancer Ther. 2012;11(12):2588-2599. DOI: 10.1158/1535-7163.MCT-12-0245

25. Miyamoto K., Khosrof S., Bursell S.E., Moromi-zato Y., Aiello L.P., Ogura Y., Adamis A.P. Vascular endothelial growth factor (VEGF)-induced retinal vascular permeability is mediated by intercellular adhesion molecule-1 (ICAM-1). Am J Pathol. 2000;156(5):1733-1739. DOI: 10.1016/S0002-9440(10)65044-4

26. Naik M.U., Stalker T.J., Brass L.F., Naik U.P. JAM-A protects from thrombosis by suppressing integrin αIIbβ3-dependent outside-in signaling in platelets. Blood. 2012;119(14):3352-3360. DOI: 10.1182/blood-2011-12-397398

27. Preliminary report on effects of photocoagulation therapy. The Diabetic Retinopathy Study Research Group. Am J Ophthalmol. 1976;81(4):383-396. DOI: 10.1016/0002-9394(76)90292-0

28. Riddell J.R., Maier P., Sass S.N., Moser M.T., Foster B.A., Gollnick S.O. Peroxiredoxin 1 Stimulates Endothelial Cell Expression of VEGF via TLR4 Dependent Activation of HIF-1a. PLoS ONE. 2012;7(11):e50394. DOI: 10.1371/journal.pone.0050394

29. Saker S., Stewart E.A., Browning A.C., Allen C.L., Amoaku W.M. The effect of hyperglycaemia on permeability and the expression of junctional complex molecules in human retinal and choroidal endothelial cells. Exp Eye Res. 2014;(121):161-167. DOI: 10.1016/j.exer.2014.02.016

30. Scholz P., Altay L., Fauser S. A Review of Subthreshold Micropulse Laser for Treatment of Macular Disorders. Adv Ther. 2017;34(7):1528-1555. DOI: 10.1007/s12325-017-0559-y

31. Schwarzer P., Kokona D., Ebneter A., Zinker-nagel M.S. Effect of Inhibition of Colony-Stimulating Factor 1 Receptor on Choroidal Neovascularization in Mice. Am J Pathol. 2020;190(2):412-425. DOI: 10.1016/j.ajpath.2019.10.011

32. Tababat-Khani P., Berglund L.M., Agardh C.D., Gomez M.F., Agardh E. Photocoagulation of human retinal pigment epithelial cells in vitro: evaluation of necrosis, apoptosis, cell migration, cell proliferation and expression of tissue repairing and cytoprotective genes. PLoS One. 2013;8(8):e70465. DOI: 10.1371/journal.pone.0070465

33. Tae N., Lee S., Kim O., Park J., Na S., Lee J.H. Syntenin promotes VEGF-induced VEGFR2 endocytosis and angiogenesis by increasing ephrin-B2 function in endothelial cells. Oncotarget. 2017;8(24):38886-38901. DOI: 10.18632/oncotarget.16452

34. Wilson A.S., Hobbs B.G., Shen W.Y., Speed T.P., Schmidt U., Begley C.G., Rakoczy P.E. Argon laser photocoagulation-induced modification of gene expression in the retina. Invest Ophthalmol Vis Sci. 2003;44(4):1426-1434. DOI: 10.1167/iovs.02-0622

35. Wu W.C., Kao Y.H., Hu P.S., Chen J.H. Geldanamycin, a HSP90 inhibitor, attenuates the hypoxia-induced vascular endothelial growth factor expression in retinal pigment epithelium cells in vitro. Exp Eye Res. 2007;85(5):721-731. DOI: 10.1016/j.exer.2007.08.005

36. Yadav N.K., Jayadev C., Rajendran A., Nagpal M. Recent developments in retinal lasers and delivery systems. Indian J Ophthalmol. 2014;62(1):50-54. DOI: 10.4103/0301-4738.126179

37. Yoshida S., Yashar B.M., Hiriyanna S., Swaroop A. Microarray analysis of gene expression in the aging human retina. Invest Ophthalmol Vis Sci. 2002;43(8):2554-2560.

38. Zeng L., Xiao Q., Chen M., Margariti A., Martin D., Ivetic A., Xu H., Mason J. et al. Vascular endothelial cell growth-activated XBP1 splicing in endothelial cells is crucial for angiogenesis. Circulation. 2013;127(16):1712-1722. DOI: 10.1161/CIRCULATIONAHA.112.001337

39. Zhu N.L., Wu L., Liu P.X., Gordon E.M., Ander-son W.F., Starnes V.A., Hall F.L. Downregulation of cyclin G1 expression by retrovirus-mediated antisense gene transfer inhibits vascular smooth muscle cell proliferation and neointima formation. Circulation. 1997;96(2):628-635. DOI: 10.1161/01.cir.96.2.628

40. Zhu X., Bai Y., Yu W., Pan C., Jin E., Song D., Xu Q., Yao Y. et al. The effects of pleiotrophin in proliferative diabetic retinopathy. PLoS One. 2015;10(1):e0115523. DOI: 10.1371/journal.pone.0115523