MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas

Verma, Nagendra; Khare, Drirh; Poe, Adam J.; Amador, Cynthia; Ghiam, Sean; Fealy, Andrew; Ebrahimi, Shaghaiegh; Shadrokh, Odelia; Song, Xue-Ying; Santiskulvong, Chintda; Mastali, Mitra; Parker, Sarah; Stotland, Aleksandr; Van Eyk, Jennifer E.; Ljubimov, Alexander V.; Saghizadeh, Mehrnoosh

doi:10.3390/cells12212524

Open AccessArticle

MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas

by

Nagendra Verma

^1,2,†

,

Drirh Khare

^1,2,3,†,

Adam J. Poe

^1,2,

Cynthia Amador

^1,2,

Sean Ghiam

^1,2,4,

Andrew Fealy

^1,2,

Shaghaiegh Ebrahimi

^1,2,

Odelia Shadrokh

^1,2,

Xue-Ying Song

⁵,

Chintda Santiskulvong

⁵,

Mitra Mastali

⁶,

Sarah Parker

⁶,

Aleksandr Stotland

⁶,

Jennifer E. Van Eyk

⁶,

Alexander V. Ljubimov

^1,2,7

and

Mehrnoosh Saghizadeh

^1,2,7,*

¹

Eye Program, Board of Governors Regenerative Medicine Institute, Cedars Sinai Medical Center, 8700 Beverly Boulevard, AHSP-A8104, Los Angeles, CA 90048, USA

²

Departments of Biomedical Sciences, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA

³

Division of Pediatric Blood and Marrow Transplantation & Cellular Therapy, University of Minnesota, Minneapolis, MN 55455, USA

⁴

Sackler School of Medicine, New York State/American Program of Tel Aviv University, Tel Aviv 6997801, Israel

⁵

Genomics Core, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA

⁶

Advanced Clinical Biosystems Research Institute, The Smidt Heart Institute, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA

⁷

Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA 90024, USA

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Cells 2023, 12(21), 2524; https://doi.org/10.3390/cells12212524

Submission received: 13 September 2023 / Revised: 8 October 2023 / Accepted: 18 October 2023 / Published: 25 October 2023

(This article belongs to the Special Issue Extracellular Vesicles: Potential Roles in Regenerative Medicine)

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Abstract

Epithelial and stromal/mesenchymal limbal stem cells contribute to corneal homeostasis and cell renewal. Extracellular vesicles (EVs), including exosomes (Exos), can be paracrine mediators of intercellular communication. Previously, we described cargos and regulatory roles of limbal stromal cell (LSC)-derived Exos in non-diabetic (N) and diabetic (DM) limbal epithelial cells (LECs). Presently, we quantify the miRNA and proteome profiles of human LEC-derived Exos and their regulatory roles in N- and DM-LSC. We revealed some miRNA and protein differences in DM vs. N-LEC-derived Exos’ cargos, including proteins involved in Exo biogenesis and packaging that may affect Exo production and ultimately cellular crosstalk and corneal function. Treatment by N-Exos, but not by DM-Exos, enhanced wound healing in cultured N-LSCs and increased proliferation rates in N and DM LSCs vs. corresponding untreated (control) cells. N-Exos-treated LSCs reduced the keratocyte markers ALDH3A1 and lumican and increased the MSC markers CD73, CD90, and CD105 vs. control LSCs. These being opposite to the changes quantified in wounded LSCs. Overall, N-LEC Exos have a more pronounced effect on LSC wound healing, proliferation, and stem cell marker expression than DM-LEC Exos. This suggests that regulatory miRNA and protein cargo differences in DM- vs. N-LEC-derived Exos could contribute to the disease state.

Keywords: diabetic cornea; extracellular vesicles; exosome; cellular crosstalk; miRNA; limbal stem cells; mesenchymal stem cells; limbal epithelial cells; proteomics; RNA-seq

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MDPI and ACS Style

Verma, N.; Khare, D.; Poe, A.J.; Amador, C.; Ghiam, S.; Fealy, A.; Ebrahimi, S.; Shadrokh, O.; Song, X.-Y.; Santiskulvong, C.; et al. MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas. Cells 2023, 12, 2524. https://doi.org/10.3390/cells12212524

AMA Style

Verma N, Khare D, Poe AJ, Amador C, Ghiam S, Fealy A, Ebrahimi S, Shadrokh O, Song X-Y, Santiskulvong C, et al. MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas. Cells. 2023; 12(21):2524. https://doi.org/10.3390/cells12212524

Chicago/Turabian Style

Verma, Nagendra, Drirh Khare, Adam J. Poe, Cynthia Amador, Sean Ghiam, Andrew Fealy, Shaghaiegh Ebrahimi, Odelia Shadrokh, Xue-Ying Song, Chintda Santiskulvong, and et al. 2023. "MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas" Cells 12, no. 21: 2524. https://doi.org/10.3390/cells12212524

APA Style

Verma, N., Khare, D., Poe, A. J., Amador, C., Ghiam, S., Fealy, A., Ebrahimi, S., Shadrokh, O., Song, X.-Y., Santiskulvong, C., Mastali, M., Parker, S., Stotland, A., Van Eyk, J. E., Ljubimov, A. V., & Saghizadeh, M. (2023). MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas. Cells, 12(21), 2524. https://doi.org/10.3390/cells12212524

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MicroRNA and Protein Cargos of Human Limbal Epithelial Cell-Derived Exosomes and Their Regulatory Roles in Limbal Stromal Cells of Diabetic and Non-Diabetic Corneas

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