NFκB-Mediated Expression of Phosphoinositide 3-Kinase δ Is Critical for Mesenchymal Transition in Retinal Pigment Epithelial Cells

Haote Han; Yanhui Yang; Zhuo Han; Luping Wang; Lijun Dong; Hui Qi; Bing Liu; Jingkui Tian; Bart Vanhaesebroeck; Andrius Kazlauskas; Guoming Zhang; Shaochong Zhang; Hetian Lei

doi:10.3390/cells12020207

NFκB-Mediated Expression of Phosphoinositide 3-Kinase δ Is Critical for Mesenchymal Transition in Retinal Pigment Epithelial Cells

Cells. 2023 Jan 4;12(2):207. doi: 10.3390/cells12020207.

Authors

Haote Han^{1

2

3}, Yanhui Yang⁴, Zhuo Han¹, Luping Wang¹, Lijun Dong⁵, Hui Qi⁵, Bing Liu^{2

3

6}, Jingkui Tian¹, Bart Vanhaesebroeck⁷, Andrius Kazlauskas⁸, Guoming Zhang⁵, Shaochong Zhang⁵, Hetian Lei⁵

Affiliations

¹ Institute of Basic Medicine and Cancer, Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 100864, China.
² Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, MA 02114, USA.
³ Department of Ophthalmology, Harvard Medical School, Boston, MA 02115, USA.
⁴ Ningxia Key Laboratory of Prevention and Control of Common Infectious Diseases, The School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750101, China.
⁵ Shenzhen Eye Hospital, Jinan University, Shenzhen Eye Institute, Shenzhen 518000, China.
⁶ Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou 510180, China.
⁷ Cancer Institute, University College London, London WC1E 6BT, UK.
⁸ Department of Ophthalmology and Visual Sciences, Department of Physiology and Biophysics, University of Illinois at Chicago, Chicago, IL 60607, USA.

Abstract

Epithelial mesenchymal transition (EMT) plays a vital role in a variety of human diseases including proliferative vitreoretinopathy (PVR), in which retinal pigment epithelial (RPE) cells play a key part. Transcriptomic analysis showed that the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway was up-regulated in human RPE cells upon treatment with transforming growth factor (TGF)-β2, a multifunctional cytokine associated with clinical PVR. Stimulation of human RPE cells with TGF-β2 induced expression of p110δ (the catalytic subunit of PI3Kδ) and activation of NFκB/p65. CRISPR-Cas9-mediated depletion of p110δ or NFκB/p65 suppressed TGF-β2-induced fibronectin expression and activation of Akt as well as migration of these cells. Intriguingly, abrogating expression of NFκB/p65 also blocked TGF-β2-induced expression of p110δ, and luciferase reporter assay indicated that TGF-β2 induced NFκB/p65 binding to the promoter of the PIK3CD that encodes p110δ. These data reveal that NFκB/p65-mediated expression of PI3Kδ is essential in human RPE cells for TGF-β2-induced EMT, uncovering hindrance of TGF-β2-induced expression of p110δ as a novel approach to inhibit PVR.

Keywords: CRISPR/Cas9; EMT; NFκB/p65; PI3Kδ; PVR; RPE; TGF-β2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Epithelial Cells / metabolism
Humans
NF-kappa B / metabolism
Phosphatidylinositol 3-Kinase / metabolism
Phosphatidylinositol 3-Kinases / metabolism
Proto-Oncogene Proteins c-akt / metabolism
Retinal Pigment Epithelium* / metabolism
Retinal Pigments / metabolism
Transforming Growth Factor beta2 / metabolism
Transforming Growth Factor beta2 / pharmacology
Vitreoretinopathy, Proliferative* / metabolism

Substances

Phosphatidylinositol 3-Kinases
Transforming Growth Factor beta2
Phosphatidylinositol 3-Kinase
Proto-Oncogene Proteins c-akt
NF-kappa B
Retinal Pigments

Grants and funding

This research was supported by the China Scholarship Council (201806320148) to H.H., the National Natural Science Foundation of China (82070989) to H.L. and Sanming Project of Medicine in Shenzhen (no. SZSM202011015) to G.Z., S.Z. and H.L.