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Oxidative stress in retinal pigment epithelium impairs stem cells: a vicious cycle in age-related macular degeneration

Abstract

Aging, chronic oxidative stress, and inflammation are major pathogenic factors in the development and progression of age-related macular degeneration (AMD) with the loss of retinal pigment epithelium (RPE). The human RPE contains a subpopulation of progenitors (i.e., RPE stem cells—RPESCs) whose role in the RPE homeostasis is under investigation. We evaluated the paracrine effects of mature RPE cells exposed to oxidative stress (H2O2) on RPESCs behavior through co-cultural, morphofunctional, and bioinformatic approaches. RPESCs showed a decline in proliferation, an increase of the senescence-associated β-galactosidase activity, the acquisition of a senescent-like secretory phenotype (SASP), and the reduction of their stemness and differentiation competencies. IL-6 and Superoxide Dismutase 2 (SOD2) seem to be key molecules in RPESCs response to oxidative stress. Our results get insight into stress-induced senescent-associated molecular mechanisms implicated in AMD pathogenesis. The presence of chronic oxidative stress in the microenvironment reduces the RPESCs abilities, inducing and/or maintaining a pro-inflammatory retinal milieu that in turn could affect AMD onset and progression.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

AMD:

Age-related macular degeneration

CRALBP:

Cellular retinaldehyde-binding protein

COX-2:

Cyclooxygenase 2

DMEM:

Dulbecco's modified eagle medium

FBS:

Fetal bovine serum

GA:

Geographic atrophy

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

H-RPE:

Human retinal pigment epithelial cells

H2O2:

Hydrogen peroxide

KLF4:

Kruppel-like factor 4

MSCs:

Mesenchymal stromal cells

MITF:

Microphthalmia-associated transcription factor

OTX2:

Orthodenticle homeobox 2

PAX6:

Paired box 6

PBS:

Phosphate-buffered saline

MEM:

Minimum essential medium eagle

PEDF:

Pigment-epithelium derived factor

RtEGM:

Retinal pigment epithelial cell growth medium

RPESCs:

Retinal pigment epithelial progenitor cells

RPE:

Retinal pigment epithelium

RPE65:

Retinal pigment epithelium-specific 65 kDa protein

SASP:

Senescence-associated secretory phenotype

SA-β-Gal:

Senescence-associated β-galactosidase

SOX2:

SRY-box transcription factor 2

SOD-2:

Superoxide dismutase 2

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Funding

This study was supported by a Grant of Università Politecnica delle Marche to Monica Mattioli-Belmonte.

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Authors

Contributions

Conceptualization: RL, MN, CM, and MMB; methodology and investigation: RL (cell culture, RT-PCR, gene analysis) and GL (Morphological investigation); writing—review and editing: all authors; supervision and funding acquisition: CM, MB, and MMB.

Corresponding author

Correspondence to Michele Nicolai.

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Lazzarini, R., Nicolai, M., Lucarini, G. et al. Oxidative stress in retinal pigment epithelium impairs stem cells: a vicious cycle in age-related macular degeneration. Mol Cell Biochem (2021). https://doi.org/10.1007/s11010-021-04258-3

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Keywords

  • Retinal pigment epithelium
  • Stem cells
  • Oxidative stress
  • Age-related macular degeneration (AMD)
  • Inflammation
  • Senescence-associated secretory phenotype (SASP)