Dataset Information

Small Extracellular Vesicles Derived from Human Trabecular Meshwork Stem Cells are distinct from TM Cells in their Proteome Cargo and Function

ABSTRACT: Glaucoma is an optic neuropathy, the second leading cause of irreversible blindness worldwide. In our earlier studies, we demonstrated the isolation and functional efficacy (wound healing and anti-oxidant potential) of small extracellular vesicles (sEV) from the adult tissue-resident stem cells of the trabecular meshwork (TMSC). This study aimed to elucidate the protein profile of sEV derived from TMSCs to correlate these functional differences for a better understanding of their potential as a cell-free therapeutic agents for primary open angle glaucoma (POAG). TM and TMSC sEV were isolated by ultracentrifugation and characterized. The protein cargo of these sEV were analyzed by Mass spectrometry. Mass spectrometry analysis identified 2802 proteins in TMSC sEV and 2848 in TM sEV. Further Maxquant analysis revealed that 511 proteins were reproducibly quantified across the samples. and 18% of the proteins were significantly altered between TMSC and 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. 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 the future.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Trabecular Meshwork Cell, Extracellular Vesicle, Stem Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Iswarya R

LAB HEAD: Gowri Priya Chidambaranathan

PROVIDER: PXD059934 | Pride | 2025-06-13

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	HUMAN_UP_20220905.fasta	Fasta
	RI_25TMSC_EXO_H_T10_1.msf	Msf
	RI_25TMSC_EXO_H_T10_1.raw	Raw
	RI_25TMSC_EXO_H_T10_2.msf	Msf
	RI_25TMSC_EXO_H_T10_2.raw	Raw

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Publications

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

Iswarya Radhakrishnan R Maheshwari Jayapal Jeya JJ Krishnadas Subbaiah S Vanniarajan Ayyasamy A Dharmalingam Kuppamuthu K Priya Chidambaranathan Gowri CG

Molecular biology reports 20250607 1

<h4>Background</h4>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 the ...[more]

PMID: 40481930

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Project description:The goal of this study was to contrast genome-wide gene expression profiles of cultured human trabecular meshwork (HTM) cells, to that of control and primary open angle glaucoma (POAG) HTM tissues. Total RNA from cultured HTM cells and HTM tissue dissected from control and POAG anterior segments fixed in RNA laterâ¢ was linearly amplified with the OvationTM Biotin RNA Amplification and Labeling System and individually hybridized to Affymetrix Human Genome U133 Plus 2.0 high density microarrays. Data analysis was performed using the GeneSpring Software 7.0. Selected genes showing significant differential expression were validated by Quantitative real-time PCR in nonamplified RNA. Cultured HTM cells retained the expression of some genes characteristic of the HTM tissue, including chitinase 3-like 1 and matrix Gla protein, but demonstrated downregulation of physiologically important genes such as myocilin. POAG HTM tissue showed relatively small changes compared to that of control donors. These changes included the statistically significant upregulation of several genes associated with inflammation and acute-phase response, including selectin-E (ELAM-I), as well as the downregulation of the antioxidants, paraoxonase 3 and ceruloplasmin. Downregulation in cultured HTM cells of genes potentially relevant for outflow pathway function highlights the importance of developing new conditions for the culture of TM cells capable of preserving the characteristics of TM cells in vivo. Comparative analysis between control and POAG tissues suggests that the upregulation of inflammation-associated genes might be involved in the progression of glaucoma. Experiment Overall Design: Total RNA from three cultured HTM cells and HTM tissue dissected from three pairs of control and two pairs of POAG anterior segments fixed in RNA laterâ¢ was linearly amplified with the OvationTM Biotin RNA Amplification and Labeling System and individually hybridized to Affymetrix Human Genome U133 Plus 2.0 high density microarrays.

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