June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
IL-33 cleaved by neutrophil proteases are more potent regulators of human retinal microvascular endothelial cells activation and angiogenesis
Author Affiliations & Notes
  • Shivantika Bisen
    Ophthalmology Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, Michigan, United States
  • Shialendra Verma
    Ophthalmology Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, Michigan, United States
  • Nikhlesh Kumar Singh
    Ophthalmology Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, Michigan, United States
  • Footnotes
    Commercial Relationships   Shivantika Bisen None; Shialendra Verma None; Nikhlesh Kumar Singh None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2069. doi:
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    • Get Citation

      Shivantika Bisen, Shialendra Verma, Nikhlesh Kumar Singh; IL-33 cleaved by neutrophil proteases are more potent regulators of human retinal microvascular endothelial cells activation and angiogenesis. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2069.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Human interleukin-33 (IL-33) is a 270 amino acid protein belonging to the IL-1 family cytokine with important roles in inflammatory diseases. Inflammatory proteases from neutrophils (Cathepsin G and Elastase) and mast cells (tryptase and chymase) regulate the activity of IL-33 by processing full-length IL-33 into mature forms. Despite recent advances in the role of IL-33 in allergic diseases, arthritis, cancer, and cardiovascular diseases, nothing is known regarding the role of these IL-33 mature forms in retinal endothelial cell signaling and angiogenesis.

Methods : Here, we first cloned the various mature forms of human IL-33, such as IL-3395-270, IL-3399-270, IL-33109-270, and IL-33112-270 into pET30a vector, and expressed these mature IL-33 constructs in E.coli strain BL21(DE-3) and purified by immobilized metal affinity chromatography (IMAC). We next evaluated the role of these recombinant mature IL-33 proteins on human retinal microvascular endothelial cells (HRMVECs) proliferation, migration, tube formation, and sprouting angiogenesis. We also investigated the role of these recombinant mature IL-33 proteins on various cellular signaling involved in retinal endothelial cell angiogenic events by 3’-mRNA sequencing and western blotting.

Results : We observed that these mature forms of IL-33 produced by inflammatory proteases potentiate HRMVEC proliferation, migration, tube formation, and sprouting angiogenesis. We also observed that these mature forms of IL-33 are more potent in inducing angiogenic signaling and endothelial cell activation. More specifically, IL-3399-270 produced by Cathepsin G (neutrophil protease) cleavage is more potent in inducing endothelial cell activation and angiogenesis.

Conclusions : Our study suggests that blockage or inhibition of IL-33 cleavage by neutrophil proteases could be useful to limit IL-33-regulated proliferative retinopathies.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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