Study of the Level of Pro- and Antiangiogenic Growth Factors in Vitreous Fluid in Neovascular Age-Related Macular Degeneration on the Background of Treatment

Purpose: to study the level of pro- and antiangiogenic growth factors in the vitreous fluid of the eyes with the neovascular form of agerelated macular degeneration (nAMD) against the background of antiangiogenic therapy.

Patients and methods. The concentration of proangiogenic (IL-8, angiogenin, TNF-α, VEGF, bFGF) and antiangiogenic (IFN-α, TGF-β IFN-γ) growth factors in the vitreous fluid of nAMD patients was determined by multiplex flow cytometry. Patients with senile cataracts without signs of AMD and cataract patients burdened with naïve (untreated) nAMD were in the comparison groups.

Results. Compared with senile cataracts, proangiogenic cytokines TNF-α were present more frequently and at a higher level in the vitreous fluid of the eyes with treated nVMD (75 % vs. 47.5 %, p < 0.05; Msr ± m: 2.4 ± 0.5 vs. 1.4 ± 0.3 pg/ml, p < 0.05), IL-8 (100 % vs. 75 %, p < 0.01; 492.9 ± 75.7 vs. 8.5 ± 1.5 pg/ml, p < 0.01), angiogenin (3822.4 ± 498.6 vs. 2820.2 ± 319.3, p < 0.01) and FGFb (58.3 % vs. 26.7 %, p < 0.05; 10.1 ± 5.9 vs. 2.7 ± 1.0, p < 0.01). In both groups of patients, antiangiogenic factors IFN-γ and TGF-β were practically not detected, but the concentration of IFN-α was significantly higher (6.4 ± 1.7 versus 4.4 ± 0.4, p < 0.01). VEGF levels in both nosological groups were almost identical: 17.5 ± 14.0 (with nAMD) versus 18.4 ± 3.2 (n/a), while VEGF was found in vitreous fluid in the cataract group significantly more often (68.2 % vs. 17 %, p < 0.01). By way of explanation, the authors suggested that since patients with nAMD had previously been administered the anti-angiogenic drug Ranibizumab, it is possible that the VEGF production was under the control of the drug, that is, suppressed or blocked.

Conclusion. Elevated levels of proangiogenic growth factors IL-8, angiogenin, TNF-α and bFGF in vitreous fluid with nAMD against the background of antiangiogenic therapy with ranibizumab suggest the presence of other, independent of VEGF, acting mechanisms for stimulating angiogenesis. The absence of antiangiogenic growth factors IFN-γ and TGF-β in the vitreous fluid allows us to think about the presence of a defect in the control and regulation of angiogenesis in nAMD. The rare detection of VEGF in combination with a significant decrease in its concentration compared to senile cataracts against the background of treatment with ranibizumab of the eyes with nAMD demonstrates targeted inhibition in practice. Against the background of treatment with angiostatic drugs, an inversion of the angiogenic phenotype occurs with the formation of new workarounds of angiogenesis, which apparently dictates the connection of drugs with other mechanisms of action. 

1. Spaide R.F., Jaffe G.J., Sarraf D., Freund K.B. Consensus Nomenclature for Reporting Neovascular Age-Related Macular Degeneration Data. Ophthalmolog. 2020 May;127(5):616-636. doi: 10.1016/j.ophtha.2019.11.004.

2. Brawn D., Michels M., Kaiser P. Ranibizumab versus Verteporfin photodynamic therapy for neovascular age-related macular degeneration: two-year results of the ANCHOR study. Ophthalmology. 2009;116:57–65. DOI: 10.1016/j.ophtha.2008.10.018

3. Rofagha S., Bhisitkul R.B., Boyer D.S. SEVEN-UP Study Group Seven-year outcomes in ranibizumab-treated patients in ANCHOR, MARINA, and HORIZON: a multicenter cohort study (SEVEN-UP). Ophthalmology. 2013;120(11):2292–2299. DOI: 10.18240/ijo.2017.01.14.

4. Holz F.G., Figueroa M.S., Bandello F. LUMINOUS, Ranibizumab treatment in treatment-naïve neovascular age-related macular degeneration. Results From LUMINOUS, a Global Real-World Study. RETINA. 2020;40:1673–1685. DOI: 10.1097/IAE.0000000000002670

5. Age-related macular degeneration. Moscow: 2017 (In Russ.). https://www.oor.ru/vracham/klrek/klinicheskie-rekomendatsii-utverzhdennyemz-rf/

6. Slakter J.S. What to do when anti-VEGF therapy fails. Retin Physician. 2010. https://www.retinalphysician.com/issues/2010/june-2010/what-to-do-when-antivegftherapy-fails

7. Amoaku W.M., Chakravarthy U., Gale R., Gavin M., Ghanchi F. Defining response to anti-VEGF therapies in neovascular AMD. Eye (Lond). 2015;29(6):721–731. DOI: 10.1038/eye.2015.48

8. Krebs I., Glittenberg C., Ansari-Shahrezaei S. Non-responders to treatment with antagonists of vascular endothelial growth factor in age related macular degeneration. Br J Ophthalmol. 2013;97:1443–1446. DOI: 10.1136/bjophthalmol-2013-303513

9. Barış M.E., Menteş J., Afrashi F. Subgroups and Features of Poor Responders to Anti-Vascular Endothelial Growth Factor Treatment in Eyes with Neovascular Age-Related Macular Degeneration Turk J Ophthalmol. 2020;50(5):275–282. DOI: 10.4274/tjo.galenos.2020.38488

10. Likhvantseva V.G., Trubilin V.N., Kapkova S.G., Gevorkyan A.S. On the question of terminology, expert criteria for assessing the effectiveness of antiangiogenic therapy and the prevalence of refractory forms of neovascular age-related macular degeneration. Оphthalmology in Russia = Oftal’mologiya. 2021;18(3):389–398 (In Russ.). DOI: 10.18008/1816-5095-2021-3-389-398

11. Shiqi Yang, Jingke Zhao, Xiaodong Sun. Resistance to anti-VEGF therapy in neovascular age-related macular degeneration: a comprehensive review. Drug Design, Development and Therapy. 2016:10 1857–1867. DOI: 10.2147/DDDT.S97653

12. Dugel P.U. HAWK and HARRIER: Phase 3, Multicenter, Randomized, DoubleMasked Trials of Brolucizumab for Neovascular Age-Related Macular Degeneration. Ophthalmology. 2020;127:72–84. DOI: 10.1016/j.ophtha.2019.04.017

13. Rajappa M., Saxena P., Kaur J. Ocular angiogenesis: mechanisms and recent advances in therapy. Adv Clin Chem. 2010;50:103–121.

14. Dreyfuss J.L., Giordano R.J., Regatieri C.V. Ocular Angiogenesis. J Ophthalmol. 2015;2015:892043. DOI: 10.1155/2015/892043

15. Casey R., Li W.W. Factors controlling ocular angiogenesis. Am J Ophthalmol. 1997;124(4):521–529. DOI: 10.1016/s0002-9394(14)70868-2

16. Distler J.H., Hirth A., Kurowska-Stolarska M., Gay R.E., Gay S., Distler O.Q. Angiogenic and angiostatic factors in the molecular control of angiogenesis. J Nucl Med. 2003;47(3):149–161.

17. de Groot H., Schmit-Eilenberger V., Kirchhof J., Augustin A.J. Dev. Angiostatic and angiogenic factors. Ophthalmol. 2010;46:1–3. DOI: 10.1159/000320005

18. Schlecht A., Vallon M., Wagner N., Ergün S., Braunger B.M. TGF-β-Neurotrophin Interactions in Heart, Retina, and Brain. Biomolecules. 2021;11(9):1360.