Cellular senescence mediates retinal ganglion cell survival regulation post-optic nerve crush injury

Abstract

Traumatic optic neuropathy refers to optic nerve (ON) injury by trauma, including explosion and traffic accident. Retinal ganglion cell (RGC) death is the critical pathological cause of irreversible visual impairment and blindness in ON injury. We previously investigated the patterns of 11 modes of cell death in mouse retina post-ON injury. Here we aimed to identify additional signalling pathways regulating RGC survival in rodents post-ON injury. RNA sequencing analysis identified the upregulation of inflammation and cellular senescence-related genes in retina post-ON injury, which were confirmed by immunoblotting and immunofluorescence analyses. Increased expression of senescence-associated β-galactosidase (SA-βgal) in RGCs and activation of microglia were also found. Transforming growth factor-β receptor type II inhibitor (LY2109761) treatment suppressed p15^Ink4b and p21^Cip1 protein and SA-βgal expression and promoted RGC survival post-ON injury with decreasing the expression of cell death markers in retina. Consistently, senolytics (dasatinib and quercetin) treatments can promote RGC survival and alleviate the reduction of ganglion cell complex thickness and pattern electroretinography activity post-ON injury with reducing SA-βgal, p15^Ink4b, p21^Cip1, microglial activation and cell death marker expression. In summary, this study revealed the activation of cellular senescence in rodent retina post-ON injury and contribute to RGC survival regulation. Targeting cellular senescence can promote RGC survival after ON injury, suggesting a potential treatment strategy for traumatic optic neuropathy.

References

[1] TanS, YaoY, YangQ, Yuan XL, CenLP, NgTK. Diversified treatment options of adult stem cells for optic neuropathies. Cell Transplant. 2022; 31:1123512.

[2] Goldenberg-CohenN, Miller NR, RepkaMX. Traumatic optic neuropathy in children and adolescents. J AAPOS. 2004; 8(1):20-27.

[3] ZhangY, KangX, WuQ, ZhengZ, YingJ, Zhang MN. Explosive eye injuries: characteristics, traumatic mechanisms, and prognostic factors for poor visual outcomes. Mil Med Res. 2023; 10(1):3.

[4] NégrelAD, Thylefors B. The global impact of eye injuries. Ophthalmic Epidemiol. 1998; 5(3):143-169.

[5] CirovicS, BholaRM, HoseDR, et al. Computer modelling study of the mechanism of optic nerve injury in blunt trauma. Br J Ophthalmol. 2006; 90(6):778-783.

[6] CarelliV, La Morgia C, Ross-CisnerosFN, SadunAA. Optic neuropathies: the tip of the neurodegeneration iceberg. Hum Mol Genet. 2017; 26(R2): R139-R150.

[7] Sánchez-MigallónMC, Nadal-NicolásFM, Jiménez-López M, Sobrado-CalvoP, Vidal-SanzM, Agudo-Barriuso M. Brain derived neurotrophic factor maintains Brn3a expression in axotomized rat retinal ganglion cells. Exp Eye Res. 2011; 92(4):260-267.

[8] ChiH, ChangHY, SangTK. Neuronal cell death mechanisms in major neurodegenerative diseases. Int J Mol Sci. 2018; 19(10):3082.

[9] YaoY, XuY, LiangJJ, Zhuang X, NgTK. Longitudinal and simultaneous profiling of 11 modes of cell death in mouse retina post-optic nerve injury. Exp Eye Res. 2022; 222:109159.

[10] CenLP, NgTK, LiangJJ, et al. Human periodontal ligament-derived stem cells promote retinal ganglion cell survival and axon regeneration after optic nerve injury. Stem Cells. 2018; 36(6):844-855.

[11] YangQ, XuY, BinX, et al. Combined treatment of human mesenchymal stem cells and green tea extract on retinal ganglion cell regeneration in rats after optic nerve injury. Exp Eye Res. 2024; 239:109787.

[12] YangY, XuC, ChenY, et al. Green tea extract ameliorates ischemia-induced retinal ganglion cell degeneration in rats. Oxid Med Cell Longev. 2019; 2019:8407206.

[13] NgTK, ChuKO, WangCC, Pang CP. Green tea catechins as therapeutic antioxidants for glaucoma treatment. Antioxidants. 2023; 12(7):1320.

[14] YuanXL, ChenSL, XuY, et al. Green tea extract enhances retinal ganglion cell survival and axonal regeneration in rats with optic nerve injury. J Nutr Biochem. 2023; 117:109333.

[15] CenLP, NgTK, LiangJJ, et al. Agonist of growth hormone-releasing hormone enhances retinal ganglion cell protection induced by macrophages after optic nerve injury. Proc Natl Acad Sci USA. 2021; 118(28):e1920834118.

[16] DennisG Jr, Sherman BT, HosackDA, et al. DAVID: database for annotation, visualization, and integrated discovery. Genome Biol. 2003; 4(5): P3.

[17] SunP, XuN, LiY, HanY. Destruction of the blood-retina barrier in diabetic retinopathy depends on angiotensin-converting enzyme-mediated TGF-β1/Smad signaling pathway activation. Int Immunopharmacol. 2020; 85:106686.

[18] QianZ, ZhengK, XuY, et al. Longitudinal in vivo evaluation of retinal ganglion cell complex layer and dendrites in mice with experimental autoimmune encephalomyelitis. Exp Eye Res. 2023; 237:109708.

[19] LiLU, ZhaoY, ZhangH. P16INK4a upregulation mediated by TBK1 induces retinal ganglion cell senescence in ischemic injury. Cell Death Dis. 2017; 8(4):e2752.

[20] XuM, Pirtskhalava T, FarrJN, et al. Senolytics improve physical function and increase lifespan in old age. Nat Med. 2018; 24(8):1246-1256.

[21] TawarayamaH, FengQ, MurayamaN, Suzuki N, NakazawaT. Cyclin-dependent kinase inhibitor 2b mediates excitotoxicity-induced death of retinal ganglion cells. Invest Ophthalmol Vis Sci. 2019; 60(13):4479-4488.

[22] FengXH, LinX, DerynckR. Smad2, Smad3 and Smad4 cooperate with Sp1 to induce p15(Ink4B) transcription in response to TGF-beta. EMBO J. 2000; 19(19):5178-5193.

[23] AcostaJC, BanitoA, WuestefeldT, et al. A complex secretory program orchestrated by the inflammasome controls paracrine senescence. Nat Cell Biol. 2013; 15(8):978-990.

[24] YasudaM, TanakaY, RyuM, TsudaS, NakazawaT. RNA sequence reveals mouse retinal transcriptome changes early after axonal injury. PLoS One. 2014; 9(3):e93258.

[25] KeuthanCJ, SchaubJA, WeiM, et al. Regional gene expression in the retina, optic nerve head, and optic nerve of mice with optic nerve crush and experimental glaucoma. Int J Mol Sci. 2023; 24(18):13719.

[26] WangAG, ChenCH, YangCW, et al. Change of gene expression profiles in the retina following optic nerve injury. Brain Res Mol Brain Res. 2002; 101(1–2):82-92.

[27] Nadal-NicolásFM, Sobrado-Calvo P, Jiménez-López M, Vidal-SanzM, Agudo-BarriusoM. Long-term effect of optic nerve axotomy on the retinal ganglion cell layer. Invest Ophthalmol Vis Sci. 2015; 56(10):6095-6112.

[28] Vidal-SanzM, Galindo-Romero C, Valiente-SorianoFJ, et al. Shared and differential retinal responses against optic nerve injury and ocular hypertension. Front Neurosci. 2017; 11:235.

[29] HeS, Sharpless NE. Senescence in health and disease. Cell. 2017; 169(6):1000-1011.

[30] ChildsBG, DurikM, BakerDJ, van Deursen JM. Cellular senescence in aging and age-related disease: from mechanisms to therapy. Nat Med. 2015; 21(12):1424-1435.

[31] Paramos-de-CarvalhoD, Martins I, CristóvãoAM, et al. Targeting senescent cells improves functional recovery after spinal cord injury. Cell Rep. 2021; 36(1):109334.

[32] DaniszewskiM, Senabouth A, LiangHH, et al. Retinal ganglion cell-specific genetic regulation in primary open-angle glaucoma. Cell Genom. 2022; 2(6):100142.

[33] CaoD, JiaoX, LiuX, et al. CDKN2B polymorphism is associated with primary open-angle glaucoma (POAG) in the afro-Caribbean population of Barbados, West Indies. PLoS One. 2012; 7(6):e39278.

[34] MoschosMM, Dettoraki M, KareklaA, et al. Polymorphism analysis of miR182 and CDKN2B genes in Greek patients with primary open angle glaucoma. PLoS One. 2020; 15(6):e0233692.

[35] NakamuraS, Kamakura T, OokuraT. Tongue epithelial KT-1 cell-cycle arrest by TGF-beta associated with induction of p21(Cip1) and p15 (Ink4b). Cytotechnology. 2009; 61(3):109-116.

[36] RochaLR, Nguyen Huu VA, Palomino La TorreC, et al. Early removal of senescent cells protects retinal ganglion cells loss in experimental ocular hypertension. Aging Cell. 2020; 19(2):e13089.

[37] ArikanS, ErsanI, KaracaT, et al. Quercetin protects the retina by reducing apoptosis due to ischemia-reperfusion injury in a rat model. Arq Bras Oftalmol. 2015; 78(2):100-104.

[38] GaoFJ, ZhangSH, XuP, et al. Quercetin declines apoptosis, ameliorates mitochondrial function and improves retinal ganglion cell survival and function in in vivo model of glaucoma in rat and retinal ganglion cell culture in vitro. Front Mol Neurosci. 2017; 10:285.

[39] GuarenteL, Sinclair DA, KroemerG. Human trials exploring anti-aging medicines. Cell Metab. 2024; 36(2):354-376.

[40] Di MiccoR, Krizhanovsky V, BakerD, D’Adda Di FagagnaF. Cellular senescence in ageing: from mechanisms to therapeutic opportunities. Nat Rev Mol Cell Biol. 2021; 22(2):75-95.

[41] LamkanfiM, DixitVM. Mechanisms and functions of inflammasomes. Cell. 2014; 157(5):1013-1022.

[42] WangF, SongY, LiuP, et al. Rapamycin suppresses neuroinflammation and protects retinal ganglion cell loss after optic nerve crush. Int Immunopharmacol. 2023; 119:110171.

RIGHTS & PERMISSIONS

2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.