Single nuclei transcriptomics of the in situ human limbal stem cell niche.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
21 Mar 2024
21 Mar 2024
Historique:
received:
06
09
2023
accepted:
15
03
2024
medline:
22
3
2024
pubmed:
22
3
2024
entrez:
22
3
2024
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 38514716
doi: 10.1038/s41598-024-57242-4
pii: 10.1038/s41598-024-57242-4
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
6749Informations de copyright
© 2024. The Author(s).
Références
Cotsarelis, G., Cheng, S. Z., Dong, G., Sun, T. T. & Lavker, R. M. Existence of slow-cycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: implications on epithelial stem cells. Cell 57, 201–209 (1989).
doi: 10.1016/0092-8674(89)90958-6
pubmed: 2702690
Di Girolamo, N. et al. Tracing the fate of limbal epithelial progenitor cells in the murine cornea. Stem Cells 33, 157–169 (2015).
doi: 10.1002/stem.1769
pubmed: 24966117
Amitai-Lange, A. et al. Lineage tracing of stem and progenitor cells of the murine corneal epithelium. Stem Cells 33, 230–239 (2015).
doi: 10.1002/stem.1840
pubmed: 25187087
Parfitt, G. J. et al. Immunofluorescence tomography of mouse ocular surface epithelial stem cells and their Niche microenvironment. Investig. Ophthalmol. Vis. Sci. 56, 7338–7344 (2015).
doi: 10.1167/iovs.15-18038
Schlötzer-Schrehardt, U. & Kruse, F. E. Identification and characterization of limbal stem cells. Exp. Eye Res. 81, 247–264 (2005).
doi: 10.1016/j.exer.2005.02.016
pubmed: 16051216
Sartaj, R. et al. Characterization of slow cycling corneal limbal epithelial cells identifies putative stem cell markers. Sci. Rep. 7, 3793 (2017).
doi: 10.1038/s41598-017-04006-y
pubmed: 28630424
pmcid: 5476663
Haagdorens, M. et al. Limbal stem cell deficiency: Current treatment options and emerging therapies. Stem Cells Int. 2016, 9798374 (2016).
doi: 10.1155/2016/9798374
pubmed: 26788074
Pellegrini, G. et al. Navigating market authorization: The path holoclar took to become the first stem cell product approved in the European Union. Stem Cells Transl Med 7, 146–154 (2018).
doi: 10.1002/sctm.17-0003
pubmed: 29280318
Joe, A. W. & Yeung, S. N. Concise review: Identifying limbal stem cells: classical concepts and new challenges. Stem Cells Transl. Med. 3, 318–322 (2014).
doi: 10.5966/sctm.2013-0137
pubmed: 24327757
Sacchetti, M., Rama, P., Bruscolini, A. & Lambiase, A. Limbal stem cell transplantation: Clinical results, limits, and perspectives. Stem Cells Int. 2018, 8086269 (2018).
doi: 10.1155/2018/8086269
pubmed: 30405723
pmcid: 6201383
Dua, H. S. & Azuara-Blanco, A. Limbal stem cells of the corneal epithelium. Surv. Ophthalmol. 44, 415–425 (2000).
doi: 10.1016/S0039-6257(00)00109-0
pubmed: 10734241
Chee, K. Y. H., Kicic, A. & Wiffen, S. J. Limbal stem cells: The search for a marker. Clin. Exp. Ophthalmol. 34, 64–73 (2006).
doi: 10.1111/j.1442-9071.2006.01147.x
pubmed: 16451261
Ebrahimi, M., Taghi-Abadi, E. & Baharvand, H. Limbal stem cells in review. J. Ophthalmic Vis. Res. 4, 40–58 (2009).
pubmed: 23056673
pmcid: 3448387
Ksander, B. R. et al. ABCB5 is a limbal stem cell gene required for corneal development and repair. Nature 511, 353–357 (2014).
doi: 10.1038/nature13426
pubmed: 25030174
pmcid: 4246512
Mikhailova, A. et al. Comparative proteomics reveals human pluripotent stem cell-derived limbal epithelial stem cells are similar to native ocular surface epithelial cells. Sci. Rep. 5, 14684 (2015).
doi: 10.1038/srep14684
pubmed: 26423138
pmcid: 4589773
Mei, H., Nakatsu, M. N., Baclagon, E. R. & Deng, S. X. Frizzled 7 maintains the undifferentiated state of human limbal stem/progenitor cells. Stem Cells 32, 938–945 (2014).
doi: 10.1002/stem.1582
pubmed: 24170316
Rama, P. et al. Limbal stem-cell therapy and long-term corneal regeneration. N. Engl. J. Med. 363, 147–155 (2010).
doi: 10.1056/NEJMoa0905955
pubmed: 20573916
Barbaro, V. et al. C/EBPδ regulates cell cycle and self-renewal of human limbal stem cells. J. Cell. Biol. 177, 1037–1049 (2007).
doi: 10.1083/jcb.200703003
pubmed: 17562792
pmcid: 2064364
Collins, P. J. et al. Epithelial chemokine CXCL14 synergizes with CXCL12 via allosteric modulation of CXCR4. FASEB J. 31, 3084–3097 (2017).
doi: 10.1096/fj.201700013R
pubmed: 28360196
pmcid: 5472405
Ligocki, A. J. et al. Molecular characteristics and spatial distribution of adult human corneal cell subtypes. Sci. Rep. 11, 16323 (2021).
doi: 10.1038/s41598-021-94933-8
pubmed: 34381080
pmcid: 8357950
Li, J. et al. S100A expression in normal corneal-limbal epithelial cells and ocular surface squamous cell carcinoma tissue. Mol. Vis. 17, 2263–2271 (2011).
pubmed: 21897749
pmcid: 3164687
Ojeda, A. F., Munjaal, R. P. & Lwigale, P. Y. Expression of CXCL12 and CXCL14 during eye development in chick and mouse. Gene Expr. Patterns 13, 303–310 (2013).
doi: 10.1016/j.gep.2013.05.006
pubmed: 23727298
Hayashi, R. et al. Coordinated generation of multiple ocular-like cell lineages and fabrication of functional corneal epithelial cell sheets from human iPS cells. Nat. Protoc. 12, 683–696 (2017).
doi: 10.1038/nprot.2017.007
pubmed: 28253236
Sagga, N., Kuffová, L., Vargesson, N., Erskine, L. & Collinson, J. M. Limbal epithelial stem cell activity and corneal epithelial cell cycle parameters in adult and aging mice. Stem Cell Res. 33, 185–198 (2018).
doi: 10.1016/j.scr.2018.11.001
pubmed: 30439642
pmcid: 6288239
Català, P. et al. Single cell transcriptomics reveals the heterogeneity of the human cornea to identify novel markers of the limbus and stroma. Sci. Rep. 11, 21727 (2021).
doi: 10.1038/s41598-021-01015-w
pubmed: 34741068
pmcid: 8571304
Collin, J. et al. A single cell atlas of human cornea that defines its development, limbal progenitor cells and their interactions with the immune cells. Ocul. Surf. 21, 279–298 (2021).
doi: 10.1016/j.jtos.2021.03.010
pubmed: 33865984
pmcid: 8343164
Chen, J. et al. Targeting matrix metalloproteases in diabetic wound healing. Front. Immunol. 14, (2023).
Li, D.-Q. et al. Single-cell transcriptomics identifies limbal stem cell population and cell types mapping its differentiation trajectory in limbal basal epithelium of human cornea. Ocul. Surf. 20, 20–32 (2021).
doi: 10.1016/j.jtos.2020.12.004
pubmed: 33388438
pmcid: 8359589
Rong, L., Wang, L., Shuai, Y., Guo, H. & Liu, K. CXCL14 regulates cell proliferation, invasion, migration and epithelial-mesenchymal transition of oral squamous cell carcinoma. Biotechnol. Biotechnol. Equip. 33, 1335–1342 (2019).
doi: 10.1080/13102818.2019.1664930
Shellenberger, T. D. et al. BRAK/CXCL14 is a potent inhibitor of angiogenesis and a chemotactic factor for immature dendritic cells. Cancer Res. 64, 8262–8270 (2004).
doi: 10.1158/0008-5472.CAN-04-2056
pubmed: 15548693
Parapuram, S. K., Huh, K., Liu, S. & Leask, A. Integrin β1 is necessary for the maintenance of corneal structural integrity. Investig. Ophthalmol. Vis. Sci. 52, 7799–7806 (2011).
doi: 10.1167/iovs.10-6945
Wolock, S. L., Lopez, R. & Klein, A. M. Scrublet: Computational identification of cell doublets in single-cell transcriptomic data. Cell Syst. 8, 281-291.e9 (2019).
doi: 10.1016/j.cels.2018.11.005
pubmed: 30954476
pmcid: 6625319
Lun, A. T. L., McCarthy, D. J. & Marioni, J. C. A step-by-step workflow for low-level analysis of single-cell RNA-seq data with bioconductor. F1000Res 5, 2122 (2016).
Stuart, T. et al. Comprehensive integration of single-cell data. Cell 177, 1888-1902.e21 (2019).
doi: 10.1016/j.cell.2019.05.031
pubmed: 31178118
pmcid: 6687398
Cao, J. et al. The single-cell transcriptional landscape of mammalian organogenesis. Nature 566, 496–502 (2019).
doi: 10.1038/s41586-019-0969-x
pubmed: 30787437
pmcid: 6434952