The ectonucleoside triphosphate diphosphohydrolase-2 (NTPDase2) in human endometrium: a novel marker of basal stroma and mesenchymal stem cells

The human endometrium undergoes repetitive regeneration cycles in order to recover the functional layer, shed during menses. The basal layer, which remains in charge of endometrial regeneration in every cycle, contains adult stem or progenitor cells of epithelial and mesenchymal lineage. Some pathologies such as adenomyosis, in which endometrial tissue develops within the myometrium, originate from this layer. It is well known that the balance between adenosine triphosphate (ATP) and adenosine plays a crucial role in stem/progenitor cell physiology, influencing proliferation, differentiation, and migration. The extracellular levels of nucleotides and nucleosides are regulated by the ectonucleotidases, such as the nucleoside triphosphate diphosphohydrolase 2 (NTPDase2). NTPDase2 is a membrane-expressed enzyme found in cells of mesenchymal origin such as perivascular cells of different tissues and the stem cells of adult neurogenic regions. The aim of this study was to characterize the expression of NTPDase2 in human nonpathological cyclic and postmenopausic endometria and in adenomyosis. We examined proliferative, secretory, and atrophic endometria from women without endometrial pathology and also adenomyotic lesions. Importantly, we identified NTPDase2 as the first marker of basal endometrium since other stromal cell markers such as CD10 label the entire stroma. As expected, NTPDase2 was also found in adenomyotic stroma, thus becoming a convenient tracer of these lesions. We did not record any changes in the expression levels or the localization of NTPDase2 along the cycle, thus suggesting that the enzyme is not influenced by the female sex hormones like other previously studied ectoenzymes. Remarkably, NTPDase2 was expressed by the Sushi Domain containing 2 (SUSD2)+ endometrial mesenchymal stem cells (eMSCs) found perivascularly, rendering it useful as a cell marker to improve the isolation of eMSCs needed for regenerative medicine therapies. Similar content being viewed by others

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Stem Cells Dev 27:1715–1728. https://doi.org/10.1089/scd.2018.0146 Acknowledgments This study was supported by a grant from the Instituto de Salud Carlos III (FIS PI15/00036), co-funded by FEDER funds/European Regional Development Fund (ERDF)-“a Way to Build Europe”-//FONDOS FEDER “una manera de hacer Europa,” and a grant from the Fundación Merck Salud (Ayuda Merck de Investigación 2016-Fertilidad). ARM was awarded a fellowship from the Asociación Española Contra el Cáncer (AECC). JS received support from the Canadian Institutes of Health Research (CIHR) and was the recipient of a “Chercheur National” research award from the Fonds de recherche du Québec – Santé (FRQS). We thank CERCA Programme (Generalitat de Catalunya) for institutional support. We are grateful to Inmaculada Gómez de Aranda for technical support and to Benjamín Torrejón of Serveis Científics I Tecnològics (Campus Bellvitge, Universitat de Barcelona). The authors thank Tom Yohannan for language editing. Funding This study was funded by Instituto de Salud Carlos III (grant number FIS PI15/00036); FEDER funds/European Regional Development Fund (ERDF)-“a Way to Build Europe”; Fundación Merck Salud (Ayuda Merck de Investigación 2016-Fertilidad). Author information Authors and Affiliations Corresponding author Ethics declarations Conflict of interest The authors declare that they have no conflict of interest. Ethical approval All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments, or comparable ethical standards. Informed consent Informed consent was obtained from all individual participants in the study. Additional information Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Electronic supplementary material Suppl. Fig. 1 Immunolocalization of NTPDase2 in human proliferative (A), secretory (B), and atrophic (C) endometrium. Stroma is labelled in the three cases although labelling is restricted to basal layer in cyclic endometria (A, B). NTPDase2 antibodies used were ALX-215-045 from Enzo (A, C) and H9s from http://ectonucleotidases-ab.com (B). Scale bars 400 μm (A), 300 μm (B), and 100 μm (C) (PNG 2943 kb) Suppl. Fig. 2 Confocal fluorescence images of some vessels of human atrophic endometrium labeled with NTPDase2 (A), CD146 (B), and PDGFRβ (C). Merged image shows a more external position of NTPDase2+ cells than CD146- and PDGFRβ-positive cells in the perivascular region (D). NTPDase2 antibody used was H9s from http://ectonucleotidases-ab.com. Scale bar 25 μm (D) (PNG 1128 kb) (JPG 216 kb) Suppl. Fig. 3 Confocal fluorescence images of endometrial blood vessels labeled with PECAM-1 (CD31) and NTPDase1 (CD39). Endothelial cells labelled with CD31 (A, E) are also positive for NTPDase1 (B, F) as shown in merge images (D, H). Scale bars 20 μm (PNG 1.01 mb) Rights and permissions About this article Cite this article Trapero, C., Vidal, A., Rodríguez-Martínez, A. et al. The ectonucleoside triphosphate diphosphohydrolase-2 (NTPDase2) in human endometrium: a novel marker of basal stroma and mesenchymal stem cells. Purinergic Signalling 15, 225–236 (2019). https://doi.org/10.1007/s11302-019-09656-3 Received: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s11302-019-09656-3