Effects of netarsudil on actin-driven cellular functions in normal and glaucomatous trabecular meshwork cells: A live imaging study

Kate E. Keller, Casey Kopczynski

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The actin cytoskeleton of trabecular meshwork (TM) cells is a therapeutic target for lowering intraocular pressure (IOP) in glaucoma patients. Netarsudil (the active ingredient in Rhopressa™) is a Rho-associated protein kinase inhibitor that induces disassembly of actin stress fibers. Here, we used live cell imaging of SiR-actin-labeled normal (NTM) and glaucomatous TM (GTM) cells to investigate actin dynamics during actin-driven biological processes with and without netarsudil treatment. Actin stress fibers were thicker in GTM than NTM cells and took longer (>120 min) to disassemble following addition of 1 µM netarsudil. Actin-rich extracellular vesicles (EVs) were derived by two mechanisms: exocytosis of intracellular-derived vesicles, and cleavage of filopodial tips, which detached the filopodia from the substratum, allowing them to retract to the cell body. While some phagocytosis was noted in untreated TM cells, netarsudil potently stimulated phagocytic uptake of EVs. Netarsudil treatment induced lateral fusion of tunneling nanotubes (TNTs) that connected adjacent TM cells; TNTs are important for TM cellular communication. Together, our results suggest that netarsudil may clear outflow channels in TM tissue by inducing phagocytosis and/or by modulating TM communication via EVs and TNTs. These cellular functions likely work together to regulate IOP in normal and glaucomatous TM.

Original languageEnglish (US)
Article number3524
Pages (from-to)1-15
Number of pages15
JournalJournal of Clinical Medicine
Volume9
Issue number11
DOIs
StatePublished - Nov 2020

Keywords

  • Actin cytoskeleton
  • Extracellular vesicles
  • Glaucoma
  • Live cell imaging
  • Trabecular meshwork
  • Tunneling nanotubes

ASJC Scopus subject areas

  • Medicine(all)

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