Synergistic Interactions of PlGF and VEGF Contribute to Blood-retinal Barrier Breakdown Through Canonical NFκB Activation

Overview

Journal Exp Cell Res

Specialty Cell Biology

Date 2020 Nov 1

PMID 33130176

Citations 8

Authors

Anton Lennikov

Anthony Mukwaya

Lijuan Fan

Madhu Sudhana Saddala

Sandro De Falco

Hu Huang

Affiliations

Soon will be listed here.

Abstract

To investigate the role of placental growth factor/vascular endothelial growth factor (PlGF-VEGF) heterodimers are involved in the blood-retinal barrier (BRB) breakdown and the associated mechanism, human retinal endothelial cells (HRECs) were treated with recombinant human (rh)PlGF-VEGF heterodimers and rhPlGF and studied in normal and high-glucose conditions. HREC barrier function was evaluated by the measurement of trans-endothelial electrical resistance (TEER). Adeno-Associated Virus Type 5 (AAV5) vectors overexpressed PlGF in the retina by intravitreal injection into the C57BL6 mouse eye. AAV5-GFP vector and naïve animals were used as controls. Immunofluorescence (IF) and western blots examined the protein expression of PlGF-VEGF heterodimers, VEGF, PlGF, NFκB, p-IκBα, ZO-1, and VE-cadherin in HREC and mouse retina. PlGF-VEGF heterodimers were detected predominantly in the HREC cell nuclei based on IF and cytoplasmic and nuclear fractionation experiments. High glucose treatment increased PlGF-VEGF nuclear abundance. Dot immunoblotting demonstrated a strong affinity of the 5D11D4 antibody to PlGF-VEGF heterodimers. rhPlGF-VEGF disrupted the barrier function of HREC, which was prevented by the neutralization of PlGF-VEGF by the 5D11D4 antibody. Stimulation of HRECs with rhPlGF also led to an increase in the nuclear signals for PlGF-VEGF, p-IκBα, and colocalization of NFκB p65 and PlGF-VEGF in the nuclei. The selective IKK2 inhibitor IMD0354 disrupted the nuclear colocalization. Treatment with IMD0354 restored the barrier function of HREC, as indicated by the ZO-1 and VE-cadherin expression. In the mouse retinas, PlGF overexpression by AAV5 vector reduced ZO-1 expression and increased abundance of pIκBα. PIGF/VEGF heterodimers mediate BRB breakdown potentially through the canonical NFκB activation.

Citing Articles

Identification of cuproptosis-related genes in septic shock based on bioinformatic analysis.

Zhao J, Zhang M, Wang Y, He F, Zhang Q PLoS One. 2024; 19(12):e0315219.

PMID: 39652607 PMC: 11627398. DOI: 10.1371/journal.pone.0315219.

Placental growth factor mediates pathological uterine angiogenesis by activating the NFAT5-SGK1 signaling axis in the endometrium: implications for preeclampsia development.

Raja Xavier J, Okumura T, Apweiler M, Chacko N, Singh Y, Brucker S Biol Res. 2024; 57(1):55.

PMID: 39152497 PMC: 11330076. DOI: 10.1186/s40659-024-00526-w.

Pathophysiology and clinical aspects of epiretinal membrane - review.

Ozog M, Nowak-Was M, Rokicki W Front Med (Lausanne). 2023; 10:1121270.

PMID: 37636571 PMC: 10447902. DOI: 10.3389/fmed.2023.1121270.

Human retinal secretome: A cross-link between mesenchymal and retinal cells.

Donato L, Scimone C, Alibrandi S, Scalinci S, Morda D, Rinaldi C World J Stem Cells. 2023; 15(7):665-686.

PMID: 37545752 PMC: 10401416. DOI: 10.4252/wjsc.v15.i7.665.

Inhibition of NF-κB ameliorates aberrant retinal glia activation and inflammatory responses in streptozotocin-induced diabetic rats.

Ding X, Sun Z, Guo Y, Tang W, Shu Q, Xu G Ann Transl Med. 2023; 11(5):197.

PMID: 37007562 PMC: 10061458. DOI: 10.21037/atm-22-2204.

References

Huang H, Van de Veire S, Dalal M, Parlier R, Semba R, Carmeliet P . Reduced retinal neovascularization, vascular permeability, and apoptosis in ischemic retinopathy in the absence of prolyl hydroxylase-1 due to the prevention of hyperoxia-induced vascular obliteration. Invest Ophthalmol Vis Sci. 2011; 52(10):7565-73. PMC: 3183980. DOI: 10.1167/iovs.11-8002. View

Saddala M, Lennikov A, Grab D, Liu G, Tang S, Huang H . Proteomics reveals ablation of PlGF increases antioxidant and neuroprotective proteins in the diabetic mouse retina. Sci Rep. 2018; 8(1):16728. PMC: 6233167. DOI: 10.1038/s41598-018-34955-x. View

Miyamoto N, De Kozak Y, Jeanny J, Glotin A, Mascarelli F, Massin P . Placental growth factor-1 and epithelial haemato-retinal barrier breakdown: potential implication in the pathogenesis of diabetic retinopathy. Diabetologia. 2006; 50(2):461-70. DOI: 10.1007/s00125-006-0539-2. View

Carmeliet P, Jain R . Molecular mechanisms and clinical applications of angiogenesis. Nature. 2011; 473(7347):298-307. PMC: 4049445. DOI: 10.1038/nature10144. View

Witmer A, Vrensen G, van Noorden C, Schlingemann R . Vascular endothelial growth factors and angiogenesis in eye disease. Prog Retin Eye Res. 2003; 22(1):1-29. DOI: 10.1016/s1350-9462(02)00043-5. View

Pfister F, Przybyt E, Harmsen M, Hammes H . Pericytes in the eye. Pflugers Arch. 2013; 465(6):789-96. DOI: 10.1007/s00424-013-1272-6. View

Dewerchin M, Carmeliet P . PlGF: a multitasking cytokine with disease-restricted activity. Cold Spring Harb Perspect Med. 2012; 2(8). PMC: 3405829. DOI: 10.1101/cshperspect.a011056. View

Carmeliet P, Moons L, Luttun A, Vincenti V, Compernolle V, de Mol M . Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions. Nat Med. 2001; 7(5):575-83. DOI: 10.1038/87904. View

Apicella I, Cicatiello V, Acampora D, Tarallo V, De Falco S . Full Functional Knockout of Placental Growth Factor by Knockin with an Inactive Variant Able to Heterodimerize with VEGF-A. Cell Rep. 2018; 23(12):3635-3646. DOI: 10.1016/j.celrep.2018.05.067. View

10.

Koizumi K, Poulaki V, Doehmen S, Welsandt G, Radetzky S, Lappas A . Contribution of TNF-alpha to leukocyte adhesion, vascular leakage, and apoptotic cell death in endotoxin-induced uveitis in vivo. Invest Ophthalmol Vis Sci. 2003; 44(5):2184-91. DOI: 10.1167/iovs.02-0589. View

11.

Snuderl M, Batista A, Kirkpatrick N, Ruiz de Almodovar C, Riedemann L, Walsh E . Targeting placental growth factor/neuropilin 1 pathway inhibits growth and spread of medulloblastoma. Cell. 2013; 152(5):1065-76. PMC: 3587980. DOI: 10.1016/j.cell.2013.01.036. View

12.

Joussen A, Poulaki V, Mitsiades N, Kirchhof B, Koizumi K, Dohmen S . Nonsteroidal anti-inflammatory drugs prevent early diabetic retinopathy via TNF-alpha suppression. FASEB J. 2002; 16(3):438-40. DOI: 10.1096/fj.01-0707fje. View

13.

Kowalczuk L, Touchard E, Omri S, Jonet L, Klein C, Valamanes F . Placental growth factor contributes to micro-vascular abnormalization and blood-retinal barrier breakdown in diabetic retinopathy. PLoS One. 2011; 6(3):e17462. PMC: 3049767. DOI: 10.1371/journal.pone.0017462. View

14.

Rosenbaum-Dekel Y, Fuchs A, Yakirevich E, Azriel A, Mazareb S, Resnick M . Nuclear localization of long-VEGF is associated with hypoxia and tumor angiogenesis. Biochem Biophys Res Commun. 2005; 332(1):271-8. DOI: 10.1016/j.bbrc.2005.04.123. View

15.

Khaliq A, Foreman D, Ahmed A, Weich H, Gregor Z, McLeod D . Increased expression of placenta growth factor in proliferative diabetic retinopathy. Lab Invest. 1998; 78(1):109-16. View

16.

Takahashi A, Sasaki H, Kim S, Tobisu K, Kakizoe T, Tsukamoto T . Markedly increased amounts of messenger RNAs for vascular endothelial growth factor and placenta growth factor in renal cell carcinoma associated with angiogenesis. Cancer Res. 1994; 54(15):4233-7. View

17.

Huang H, Lennikov A, Saddala M, Gozal D, Grab D, Khalyfa A . Placental growth factor negatively regulates retinal endothelial cell barrier function through suppression of glucose-6-phosphate dehydrogenase and antioxidant defense systems. FASEB J. 2019; 33(12):13695-13709. PMC: 6894064. DOI: 10.1096/fj.201901353R. View

18.

Green C, Lichtlen P, Huynh N, Yanovsky M, Laderoute K, Schaffner W . Placenta growth factor gene expression is induced by hypoxia in fibroblasts: a central role for metal transcription factor-1. Cancer Res. 2001; 61(6):2696-703. View

19.

Wells J, Glassman A, Ayala A, Jampol L, Bressler N, Bressler S . Aflibercept, Bevacizumab, or Ranibizumab for Diabetic Macular Edema: Two-Year Results from a Comparative Effectiveness Randomized Clinical Trial. Ophthalmology. 2016; 123(6):1351-9. PMC: 4877252. DOI: 10.1016/j.ophtha.2016.02.022. View

20.

Beltramo E, Porta M . Pericyte loss in diabetic retinopathy: mechanisms and consequences. Curr Med Chem. 2013; 20(26):3218-25. DOI: 10.2174/09298673113209990022. View