Unravelling the interplay of sphingolipids and TGF-β signaling in the human corneal stroma

Sarah E. Nicholas, Tyler G. Rowsey, Shrestha Priyadarsini, Nawajes Mandal, Dimitrios Karamichos

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Purpose: To delineate the role of Sphingolipids (SPLs) in the human cornea and their cross-talks with transforming growth factor beta (TGF-β) in order to develop novel, non-invasive therapies. Methods: Human corneal fibroblasts (HCFs) were harvested from healthy donors, stimulated with Vitamin C to promote extracellular matrix assembly, treated with exogenous sphingosine-1phosphate (S1P) or sphingosine kinase inhibitor 2 (SPHK I2) and isolated after 4 weeks for further analysis. Results: Data showed that S1P led to a significant decrease in cellular migration where SPHK I2 just delayed it for 24h. Significant modulation of the sphingolipid pathway was also noted. Sphingosine kinase-1 (SphK1) was significantly downregulated upon exogenous stimulation with S1P at a concentration of 5μM and Sphingosine kinase-2 (SphK2) was also significantly downregulated at concentrations of 0.01μM, 0.1μM, and 5μM; whereas no effects were observed upon stimulation with SPHK I2. S1PR3 was significantly downregulated by 0.1μM and 5μM S1P and upregulated by 5μM and 10μM SPHK I2. Furthermore, both S1P and SPHK I2 regulated corneal fibrosis markers such as alpha-smooth muscle actin, collagen I, III, and V. We also investigated the interplay between two TGF-β isoforms and S1P/SPHK I2 treatments and found that TGF-β1 and TGF-β3 were both significantly upregulated with the 0.1μM S1P but were significantly downregulated with the 5μM S1P concentration. When TGF-β1 was compared directly to TGF-β3 expression, we observed that TGF-β3 was significantly downregulated compared to TGF-β1 in the 5μM concentration of S1P. No changes were observed upon SPHK I2 treatment. Conclusion: Our study delineates the role of sphingolipids in the human cornea and highlights their different activities based on the cell/tissue type.

Original languageEnglish (US)
Article numbere0182390
JournalPLoS ONE
Volume12
Issue number8
DOIs
StatePublished - Aug 1 2017

Fingerprint

Corneal Stroma
sphingosine
sphingolipids
Sphingolipids
Sphingosine
transforming growth factor beta
Transforming Growth Factor beta
Down-Regulation
phosphotransferases (kinases)
transforming growth factor beta 3
transforming growth factor beta 1
Cornea
sphingosine kinase
cornea
Fibroblasts
Ascorbic Acid
Extracellular Matrix
Smooth Muscle
Muscle
Actins

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Unravelling the interplay of sphingolipids and TGF-β signaling in the human corneal stroma. / Nicholas, Sarah E.; Rowsey, Tyler G.; Priyadarsini, Shrestha; Mandal, Nawajes; Karamichos, Dimitrios.

In: PLoS ONE, Vol. 12, No. 8, e0182390, 01.08.2017.

Research output: Contribution to journalArticle

Nicholas, Sarah E. ; Rowsey, Tyler G. ; Priyadarsini, Shrestha ; Mandal, Nawajes ; Karamichos, Dimitrios. / Unravelling the interplay of sphingolipids and TGF-β signaling in the human corneal stroma. In: PLoS ONE. 2017 ; Vol. 12, No. 8.
@article{775ddd0e0da84044b38913c48f69f6f7,
title = "Unravelling the interplay of sphingolipids and TGF-β signaling in the human corneal stroma",
abstract = "Purpose: To delineate the role of Sphingolipids (SPLs) in the human cornea and their cross-talks with transforming growth factor beta (TGF-β) in order to develop novel, non-invasive therapies. Methods: Human corneal fibroblasts (HCFs) were harvested from healthy donors, stimulated with Vitamin C to promote extracellular matrix assembly, treated with exogenous sphingosine-1phosphate (S1P) or sphingosine kinase inhibitor 2 (SPHK I2) and isolated after 4 weeks for further analysis. Results: Data showed that S1P led to a significant decrease in cellular migration where SPHK I2 just delayed it for 24h. Significant modulation of the sphingolipid pathway was also noted. Sphingosine kinase-1 (SphK1) was significantly downregulated upon exogenous stimulation with S1P at a concentration of 5μM and Sphingosine kinase-2 (SphK2) was also significantly downregulated at concentrations of 0.01μM, 0.1μM, and 5μM; whereas no effects were observed upon stimulation with SPHK I2. S1PR3 was significantly downregulated by 0.1μM and 5μM S1P and upregulated by 5μM and 10μM SPHK I2. Furthermore, both S1P and SPHK I2 regulated corneal fibrosis markers such as alpha-smooth muscle actin, collagen I, III, and V. We also investigated the interplay between two TGF-β isoforms and S1P/SPHK I2 treatments and found that TGF-β1 and TGF-β3 were both significantly upregulated with the 0.1μM S1P but were significantly downregulated with the 5μM S1P concentration. When TGF-β1 was compared directly to TGF-β3 expression, we observed that TGF-β3 was significantly downregulated compared to TGF-β1 in the 5μM concentration of S1P. No changes were observed upon SPHK I2 treatment. Conclusion: Our study delineates the role of sphingolipids in the human cornea and highlights their different activities based on the cell/tissue type.",
author = "Nicholas, {Sarah E.} and Rowsey, {Tyler G.} and Shrestha Priyadarsini and Nawajes Mandal and Dimitrios Karamichos",
year = "2017",
month = "8",
day = "1",
doi = "10.1371/journal.pone.0182390",
language = "English (US)",
volume = "12",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "8",

}

TY - JOUR

T1 - Unravelling the interplay of sphingolipids and TGF-β signaling in the human corneal stroma

AU - Nicholas, Sarah E.

AU - Rowsey, Tyler G.

AU - Priyadarsini, Shrestha

AU - Mandal, Nawajes

AU - Karamichos, Dimitrios

PY - 2017/8/1

Y1 - 2017/8/1

N2 - Purpose: To delineate the role of Sphingolipids (SPLs) in the human cornea and their cross-talks with transforming growth factor beta (TGF-β) in order to develop novel, non-invasive therapies. Methods: Human corneal fibroblasts (HCFs) were harvested from healthy donors, stimulated with Vitamin C to promote extracellular matrix assembly, treated with exogenous sphingosine-1phosphate (S1P) or sphingosine kinase inhibitor 2 (SPHK I2) and isolated after 4 weeks for further analysis. Results: Data showed that S1P led to a significant decrease in cellular migration where SPHK I2 just delayed it for 24h. Significant modulation of the sphingolipid pathway was also noted. Sphingosine kinase-1 (SphK1) was significantly downregulated upon exogenous stimulation with S1P at a concentration of 5μM and Sphingosine kinase-2 (SphK2) was also significantly downregulated at concentrations of 0.01μM, 0.1μM, and 5μM; whereas no effects were observed upon stimulation with SPHK I2. S1PR3 was significantly downregulated by 0.1μM and 5μM S1P and upregulated by 5μM and 10μM SPHK I2. Furthermore, both S1P and SPHK I2 regulated corneal fibrosis markers such as alpha-smooth muscle actin, collagen I, III, and V. We also investigated the interplay between two TGF-β isoforms and S1P/SPHK I2 treatments and found that TGF-β1 and TGF-β3 were both significantly upregulated with the 0.1μM S1P but were significantly downregulated with the 5μM S1P concentration. When TGF-β1 was compared directly to TGF-β3 expression, we observed that TGF-β3 was significantly downregulated compared to TGF-β1 in the 5μM concentration of S1P. No changes were observed upon SPHK I2 treatment. Conclusion: Our study delineates the role of sphingolipids in the human cornea and highlights their different activities based on the cell/tissue type.

AB - Purpose: To delineate the role of Sphingolipids (SPLs) in the human cornea and their cross-talks with transforming growth factor beta (TGF-β) in order to develop novel, non-invasive therapies. Methods: Human corneal fibroblasts (HCFs) were harvested from healthy donors, stimulated with Vitamin C to promote extracellular matrix assembly, treated with exogenous sphingosine-1phosphate (S1P) or sphingosine kinase inhibitor 2 (SPHK I2) and isolated after 4 weeks for further analysis. Results: Data showed that S1P led to a significant decrease in cellular migration where SPHK I2 just delayed it for 24h. Significant modulation of the sphingolipid pathway was also noted. Sphingosine kinase-1 (SphK1) was significantly downregulated upon exogenous stimulation with S1P at a concentration of 5μM and Sphingosine kinase-2 (SphK2) was also significantly downregulated at concentrations of 0.01μM, 0.1μM, and 5μM; whereas no effects were observed upon stimulation with SPHK I2. S1PR3 was significantly downregulated by 0.1μM and 5μM S1P and upregulated by 5μM and 10μM SPHK I2. Furthermore, both S1P and SPHK I2 regulated corneal fibrosis markers such as alpha-smooth muscle actin, collagen I, III, and V. We also investigated the interplay between two TGF-β isoforms and S1P/SPHK I2 treatments and found that TGF-β1 and TGF-β3 were both significantly upregulated with the 0.1μM S1P but were significantly downregulated with the 5μM S1P concentration. When TGF-β1 was compared directly to TGF-β3 expression, we observed that TGF-β3 was significantly downregulated compared to TGF-β1 in the 5μM concentration of S1P. No changes were observed upon SPHK I2 treatment. Conclusion: Our study delineates the role of sphingolipids in the human cornea and highlights their different activities based on the cell/tissue type.

UR - http://www.scopus.com/inward/record.url?scp=85027434924&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85027434924&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0182390

DO - 10.1371/journal.pone.0182390

M3 - Article

VL - 12

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 8

M1 - e0182390

ER -