Single nuclei transcriptomics of the in situ human limbal stem cell niche

Kathryn C. Davidson; Minkyung Sung; Karl D. Brown; Julian Contet; Serena Belluschi; Regan Hamel; Aida Moreno-Moral; Rodrigo L. dos Santos; Julian Gough; Jose M. Polo; Mark Daniell; Geraint J. Parfitt

doi:10.1038/s41598-024-57242-4

Single nuclei transcriptomics of the in situ human limbal stem cell niche

Kathryn C. Davidson, Minkyung Sung, Karl D. Brown, Julian Contet, Serena Belluschi, Regan Hamel, Aida Moreno-Moral, Rodrigo L. dos Santos, Julian Gough, Jose M. Polo, Mark Daniell, Geraint J. Parfitt

Research output: Contribution to journal › Article › Research › peer-review

8 Citations (Scopus)

Abstract

The corneal epithelium acts as a barrier to pathogens entering the eye; corneal epithelial cells are continuously renewed by uni-potent, quiescent limbal stem cells (LSCs) located at the limbus, where the cornea transitions to conjunctiva. There has yet to be a consensus on LSC markers and their transcriptome profile is not fully understood, which may be due to using cadaveric tissue without an intact stem cell niche for transcriptomics. In this study, we addressed this problem by using single nuclei RNA sequencing (snRNAseq) on healthy human limbal tissue that was immediately snap-frozen after excision from patients undergoing cataract surgery. We identified the quiescent LSCs as a sub-population of corneal epithelial cells with a low level of total transcript counts. Moreover, TP63, KRT15, CXCL14, and ITGβ4 were found to be highly expressed in LSCs and transiently amplifying cells (TACs), which constitute the corneal epithelial progenitor populations at the limbus. The surface markers SLC6A6 and ITGβ4 could be used to enrich human corneal epithelial cell progenitors, which were also found to specifically express the putative limbal progenitor cell markers MMP10 and AC093496.1.

Original language	English
Article number	6749
Number of pages	12
Journal	Scientific Reports
Volume	14
Issue number	1
DOIs	https://doi.org/10.1038/s41598-024-57242-4
Publication status	Published - Dec 2024

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Access to Document

10.1038/s41598-024-57242-4Licence: CC BY

Cite this

@article{64cc3ffc66084d268f43dc9216a11473,

title = "Single nuclei transcriptomics of the in situ human limbal stem cell niche",

abstract = "The corneal epithelium acts as a barrier to pathogens entering the eye; corneal epithelial cells are continuously renewed by uni-potent, quiescent limbal stem cells (LSCs) located at the limbus, where the cornea transitions to conjunctiva. There has yet to be a consensus on LSC markers and their transcriptome profile is not fully understood, which may be due to using cadaveric tissue without an intact stem cell niche for transcriptomics. In this study, we addressed this problem by using single nuclei RNA sequencing (snRNAseq) on healthy human limbal tissue that was immediately snap-frozen after excision from patients undergoing cataract surgery. We identified the quiescent LSCs as a sub-population of corneal epithelial cells with a low level of total transcript counts. Moreover, TP63, KRT15, CXCL14, and ITGβ4 were found to be highly expressed in LSCs and transiently amplifying cells (TACs), which constitute the corneal epithelial progenitor populations at the limbus. The surface markers SLC6A6 and ITGβ4 could be used to enrich human corneal epithelial cell progenitors, which were also found to specifically express the putative limbal progenitor cell markers MMP10 and AC093496.1.",

author = "Davidson, \{Kathryn C.\} and Minkyung Sung and Brown, \{Karl D.\} and Julian Contet and Serena Belluschi and Regan Hamel and Aida Moreno-Moral and \{dos Santos\}, \{Rodrigo L.\} and Julian Gough and Polo, \{Jose M.\} and Mark Daniell and Parfitt, \{Geraint J.\}",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2024.",

year = "2024",

month = dec,

doi = "10.1038/s41598-024-57242-4",

language = "English",

volume = "14",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Single nuclei transcriptomics of the in situ human limbal stem cell niche

AU - Davidson, Kathryn C.

AU - Sung, Minkyung

AU - Brown, Karl D.

AU - Contet, Julian

AU - Belluschi, Serena

AU - Hamel, Regan

AU - Moreno-Moral, Aida

AU - dos Santos, Rodrigo L.

AU - Gough, Julian

AU - Polo, Jose M.

AU - Daniell, Mark

AU - Parfitt, Geraint J.

PY - 2024/12

Y1 - 2024/12

N2 - The corneal epithelium acts as a barrier to pathogens entering the eye; corneal epithelial cells are continuously renewed by uni-potent, quiescent limbal stem cells (LSCs) located at the limbus, where the cornea transitions to conjunctiva. There has yet to be a consensus on LSC markers and their transcriptome profile is not fully understood, which may be due to using cadaveric tissue without an intact stem cell niche for transcriptomics. In this study, we addressed this problem by using single nuclei RNA sequencing (snRNAseq) on healthy human limbal tissue that was immediately snap-frozen after excision from patients undergoing cataract surgery. We identified the quiescent LSCs as a sub-population of corneal epithelial cells with a low level of total transcript counts. Moreover, TP63, KRT15, CXCL14, and ITGβ4 were found to be highly expressed in LSCs and transiently amplifying cells (TACs), which constitute the corneal epithelial progenitor populations at the limbus. The surface markers SLC6A6 and ITGβ4 could be used to enrich human corneal epithelial cell progenitors, which were also found to specifically express the putative limbal progenitor cell markers MMP10 and AC093496.1.

AB - The corneal epithelium acts as a barrier to pathogens entering the eye; corneal epithelial cells are continuously renewed by uni-potent, quiescent limbal stem cells (LSCs) located at the limbus, where the cornea transitions to conjunctiva. There has yet to be a consensus on LSC markers and their transcriptome profile is not fully understood, which may be due to using cadaveric tissue without an intact stem cell niche for transcriptomics. In this study, we addressed this problem by using single nuclei RNA sequencing (snRNAseq) on healthy human limbal tissue that was immediately snap-frozen after excision from patients undergoing cataract surgery. We identified the quiescent LSCs as a sub-population of corneal epithelial cells with a low level of total transcript counts. Moreover, TP63, KRT15, CXCL14, and ITGβ4 were found to be highly expressed in LSCs and transiently amplifying cells (TACs), which constitute the corneal epithelial progenitor populations at the limbus. The surface markers SLC6A6 and ITGβ4 could be used to enrich human corneal epithelial cell progenitors, which were also found to specifically express the putative limbal progenitor cell markers MMP10 and AC093496.1.

UR - https://www.scopus.com/pages/publications/85188243669

U2 - 10.1038/s41598-024-57242-4

DO - 10.1038/s41598-024-57242-4

M3 - Article

C2 - 38514716

AN - SCOPUS:85188243669

SN - 2045-2322

VL - 14

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 6749

ER -

Single nuclei transcriptomics of the in situ human limbal stem cell niche

Abstract

UN SDGs

Access to Document

Other files and links

Cite this