Research Paper Volume 16, Issue 8 pp 6673—6693

Senescent characteristics of human corneal endothelial cells upon ultraviolet-A exposure

Kohsaku Numa1,2, , Sandip Kumar Patel1,3, , Zhixin A. Zhang1, , Jordan B. Burton1, , Akifumi Matsumoto2, , Jun-Wei B. Hughes1, , Chie Sotozono2, , Birgit Schilling1, , Pierre-Yves Desprez1,4, , Judith Campisi1, , Koji Kitazawa1,2, ,

  • 1 Buck Institute for Research on Aging, Novato, CA 94945, USA
  • 2 Kyoto Prefectural University of Medicine, Department of Ophthalmology, Kyoto 6020841, Japan
  • 3 MRC Toxicology Unit, University of Cambridge, Cambridge CB2 1QR, UK
  • 4 California Pacific Medical Center, Research Institute, San Francisco, CA 94107, USA

Received: December 20, 2023       Accepted: March 28, 2024       Published: April 26, 2024
How to Cite

Copyright: © 2024 Numa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Purpose: The objective of this study was to investigate the senescent phenotypes of human corneal endothelial cells (hCEnCs) upon treatment with ultraviolet (UV)-A.

Methods: We assessed cell morphology, senescence-associated β-galactosidase (SA-β-gal) activity, cell proliferation and expression of senescence markers (p16 and p21) in hCEnCs exposed to UV-A radiation, and senescent hCEnCs induced by ionizing radiation (IR) were used as positive controls. We performed RNA sequencing and proteomics analyses to compare gene and protein expression profiles between UV-A- and IR-induced senescent hCEnCs, and we also compared the results to non-senescent hCEnCs.

Results: Cells exposed to 5 J/cm2 of UV-A or to IR exhibited typical senescent phenotypes, including enlargement, increased SA-β-gal activity, decreased cell proliferation and elevated expression of p16 and p21. RNA-Seq analysis revealed that 83.9% of the genes significantly upregulated and 82.6% of the genes significantly downregulated in UV-A-induced senescent hCEnCs overlapped with the genes regulated in IR-induced senescent hCEnCs. Proteomics also revealed that 93.8% of the proteins significantly upregulated in UV-A-induced senescent hCEnCs overlapped with those induced by IR. In proteomics analyses, senescent hCEnCs induced by UV-A exhibited elevated expression levels of several factors part of the senescence-associated secretory phenotype.

Conclusions: In this study, where senescence was induced by UV-A, a more physiological stress for hCEnCs compared to IR, we determined that UV-A modulated the expression of many genes and proteins typically altered upon IR treatment, a more conventional method of senescence induction, even though UV-A also modulated specific pathways unrelated to IR.


ACTB: beta actin; BGN: biglycan; BMP: bone morphogenetic proteins; CCL: c-c motif chemokine ligand; CCND: cyclin D; CDK: cyclin dependent kinase; CDKN: cyclin dependent kinase inhibitor; CEnCs: corneal endothelial cells; CF: complement factor; CLU: clusterin; CM: conditioned medium; COL1A1: collagen type I alpha 1 chain; COL4A5: collagen type IV alpha 5 chain; COL4A6: collagen type IV alpha 6 chain; COL6A2: collagen type VI alpha 2 chain; COL8A1: collagen type VIII alpha 1 chain; COL12A1: collagen type XII alpha 1 chain; COL17A1: collagen type XVII alpha 1 chain; CXCL: c-x-c motif chemokine ligand; DAPI: 4’,6-diamidino-2-phenylindole; DEG: differentially expressed gene; DIA: data-independent acquisition; ECM: extracellular matrix; EdU: 5-ethynyl-2’-deoxyuridine; ENO1: enolase1; FECD: Fuchs’ endothelial corneal dystrophy; FN1: fibronectin1; GDF15: growth differentiation factor 15; GO: gene ontology; GPI: glucose-6-phosphate isomerase; HMGB1: high mobility group box 1; ICAM1: intercellular adhesion molecule-1; IGF: insulin like growth factor; LDH: lactate dehydrogenase; IL: interleukin; IR: ionizing radiation; LMNB1: lamin B1; LOXL: lysyl oxidase like; LTBP2: latent-transforming growth factor beta binding protein 2; MATN: matrilin; MMP: matrix metalloproteinase; MS: mass spectrometry; NOG: noggin; NOX: NADPH oxidase; PDGFA: platelet derived growth factor subunit A; PKM: pyruvate kinase; PLS-DA: partial least squares discriminant analysis; ROS: reactive oxygen species; qRT-PCR: quantitative reverse transcription polymerase chain reaction; SA-β-Gal: senescence-associated β-galactosidase; SASP: senescence-associated secretory phenotype; SEMA: semaphorin; SERPIN: serine proteinase inhibitor; SLC2A1: solute carrier family 2 member 1; SOD: superoxide dismutase; STC1: stanniocalcin 1; TGFB: transforming growth factor beta; TGFBI: transforming growth factor beta induced protein; TPI1: triosephosphate isomerase 1; UV: ultra violet.