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Background <p>Glaucoma, an optic neuropathy, is the second leading cause of irreversible blindness worldwide. Wound healing efficiency and anti-oxidant potential of the small extracellular vesicles (sEV) isolated from the adult tissue-resident stem cells of the trabecular meshwork (TMSC) compared to trabecular meshwork (TM) cells have been demonstrated. This study aimed to compare the protein profile of extracellular vesicles derived from TMSCs and TM cells for a better understanding of their potential as a cell-free therapeutic agent for primary open angle glaucoma (POAG).</p> Methods and results <p>Proteins were isolated from TM and TMSC sEV samples (<i>n</i> = 3 each) and proteomic profiling was carried out by mass spectrometry. Mass spectrometry analysis identified 2802 proteins in TMSC sEV and 2848 in TM sEV. Differential expression analysis identified distinct protein profiles between TMSC and TM sEV. Notably, sEV from TMSCs were enriched with proteins associated with wound healing, cell proliferation, migration, anti-oxidant, and anti-apoptotic activities, consistent with the findings in other mesenchymal stem cells. Pathway analysis highlighted the enrichment of proteins associated with PI3K-AKT and MAPK signaling pathways. Further validation by western blotting confirmed that TMSC sEV effectively modulated these pathways in TM cells, which are essential for cell proliferation and survival under oxidative stress.</p> Conclusion <p>To our knowledge, this is the first comprehensive protein profiling of TMSC-derived sEV, demonstrating the presence of functional proteins and their capacity to regulate the MAPK and PI3K-AKT signaling pathways in the recipient TM cells. This highlights the potential of TMSC sEV in advancing cell-free therapeutic strategies for POAG in future.</p>

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Proteome cargo and functional heterogeneity of extracellular vesicles derived from stem cells and differentiated cells of human trabecular meshwork

  • 期刊论文

  • 发表日期：

    2025年6月7日

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  • Radhakrishnan Iswarya,
  • Jayapal Jeya Maheshwari,
  • Subbaiah Krishnadas,
  • Ayyasamy Vanniarajan,
  • Kuppamuthu Dharmalingam,
  • Chidambaranathan Gowri Priya

摘要

Background

Glaucoma, an optic neuropathy, is the second leading cause of irreversible blindness worldwide. Wound healing efficiency and anti-oxidant potential of the small extracellular vesicles (sEV) isolated from the adult tissue-resident stem cells of the trabecular meshwork (TMSC) compared to trabecular meshwork (TM) cells have been demonstrated. This study aimed to compare the protein profile of extracellular vesicles derived from TMSCs and TM cells for a better understanding of their potential as a cell-free therapeutic agent for primary open angle glaucoma (POAG).

Methods and results

Proteins were isolated from TM and TMSC sEV samples (n = 3 each) and proteomic profiling was carried out by mass spectrometry. Mass spectrometry analysis identified 2802 proteins in TMSC sEV and 2848 in TM sEV. Differential expression analysis identified distinct protein profiles between TMSC and TM sEV. Notably, sEV from TMSCs were enriched with proteins associated with wound healing, cell proliferation, migration, anti-oxidant, and anti-apoptotic activities, consistent with the findings in other mesenchymal stem cells. Pathway analysis highlighted the enrichment of proteins associated with PI3K-AKT and MAPK signaling pathways. Further validation by western blotting confirmed that TMSC sEV effectively modulated these pathways in TM cells, which are essential for cell proliferation and survival under oxidative stress.

Conclusion

To our knowledge, this is the first comprehensive protein profiling of TMSC-derived sEV, demonstrating the presence of functional proteins and their capacity to regulate the MAPK and PI3K-AKT signaling pathways in the recipient TM cells. This highlights the potential of TMSC sEV in advancing cell-free therapeutic strategies for POAG in future.