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The existence of senescent cells in conjunctival epithelium from elderly individuals

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Abstract

Purpose

The ocular surface microenvironment changes with aging. However, it remains unclear if cellular senescence influences the ocular surface. We investigated the presence of p16INK4a-expressing senescent cells in healthy human conjunctiva.

Study design

Clinical and experimental.

Methods

Healthy conjunctival tissue samples were obtained from middle-aged and elderly subjects. RT-qPCR was performed to assess the expression of senescence markers CDKN2A (p16INK4a) and CDKN1A (p21CIP1/WAF1) and immunostaining was performed to examine the expression of the senescence marker p16INK4a, stem cell markers Ki67 and p63, tight-junction marker ZO-1.

Results

Our study involved 19 conjunctival tissue samples (10 elderly and 9 middle-aged), mean age [elderly: 75.8 ± 3.7 years (72–81), middle-aged: 52.7 ± 7 years (38–59)], sex (elderly: 3 men, 7 women; middle-aged: 3 men, 6 women). The expression of p16INK4a was significantly increased at the RNA level in the elderly compared to middle-aged (p < 0.05). Positivity rate of p16INK4a was significantly elevated in the elderly (15.0 ± 7.8%) compared to middle-aged (0.2 ± 0.6%) (p < 0.05). Positivity rate of Ki67and p63 was significantly reduced in the elderly (1.7 ± 1.7% and 16.5 ± 9.5%) compared to middle-aged (3.9 ± 1.8% and 24.7 ± 5.7%) (p < 0.05). ZO-1 expression was reduced in tissue samples showing p16INK4a-positivity but retained in tissue samples in which p16INK4a was undetectable.

Conclusions

Senescent cells accumulate with age in the conjunctival epithelium, accompanied by a decrease in Ki67, p63 and ZO-1 expressing cells.

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Acknowledgements

We wish to thank Shigeru Kinoshita for the scientific suggestions. This research was partially supported by JSPS Grant-in-Aid for Scientific Research Grant Number 21K09748.

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Authors and Affiliations

  1. Department of Ophthalmology, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Hirokoji-agaru, Kawaramachi-dori, Kamigyo-ku, Kyoto, 602-0841, Japan

    Yasufumi Tomioka, Koji Kitazawa, Kohsaku Numa, Norihiko Yokoi & Chie Sotozono

  2. Buck Institute for Research on Aging, Novato, CA, 94945, USA

    Koji Kitazawa, Kohsaku Numa & Jun-Wei B. Hughes

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  1. Yasufumi Tomioka
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  2. Koji Kitazawa
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  5. Norihiko Yokoi
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  6. Chie Sotozono
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Corresponding author

Correspondence to Koji Kitazawa.

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Conflicts of interest

Y. Tomioka, None; K. Kitazawa, Seminars (AMO, Santen, Senju); K. Numa, None; J-W B. Hughes, None; N. Yokoi, Seminars (Lumenis Be, Santen, Senju), Patents (planned, pending or issued) (Rexxam and Kyoto Prefectural Public University Corporation owe the patent for an ophthalmic device related to the method of dry eye classification, Kowa and Kyoto Prefectural Public University Corporation owe the patent for the method and device for evaluating tear film dynamics); C. Sotozono, Grant to the author’s institution (Santen, Alcon, Senju, Otsuka, Nitto Medic, Nitten, Kowa, CorneaGen), Seminars (Otsuka, Santen, Senju, TOA).

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Corresponding Author: Koji Kitazawa

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Tomioka, Y., Kitazawa, K., Numa, K. et al. The existence of senescent cells in conjunctival epithelium from elderly individuals. Jpn J Ophthalmol 68, 157–165 (2024). https://doi.org/10.1007/s10384-023-01047-x

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  • DOI: https://doi.org/10.1007/s10384-023-01047-x

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