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Single-Cell RNA Sequencing Unravels Heterogeneity of the Stromal Niche in Cutaneous Melanoma Heterogeneous Spheroids

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    0539639 - ÚMG 2021 RIV CH eng J - Journal Article
    Novotný, Jiří - Strnadová, K. - Dvořánková, B. - Kocourková, Šárka - Jaksa, R. - Dundr, P. - Pačes, Václav - Smetana, K. - Kolář, Michal - Lacina, L.
    Single-Cell RNA Sequencing Unravels Heterogeneity of the Stromal Niche in Cutaneous Melanoma Heterogeneous Spheroids.
    Cancers (Basel). Roč. 12, č. 11 (2020), č. článku 3324. E-ISSN 2072-6694
    R&D Projects: GA MŠMT(CZ) LM2018131
    Institutional support: RVO:68378050
    Keywords : melanoma * spheroids * fibroblasts * subpopulation * heterogeneity * Interleukin-6 * cytokine * senescence-associated secretory phenotype * extracellular matrix * single-cell sequencing
    OECD category: Biochemistry and molecular biology
    Impact factor: 6.639, year: 2020
    Method of publishing: Open access
    https://www.mdpi.com/2072-6694/12/11/3324

    Heterogeneous spheroids have recently acquired a prominent position in melanoma research because they incorporate microenvironmental cues relevant for melanoma. In this study, we focused on the analysis of microenvironmental factors introduced in melanoma heterogeneous spheroids by different dermal fibroblasts. We aimed to map the fibroblast diversity resulting from previously acquired damage caused by exposure to extrinsic and intrinsic stimuli. To construct heterogeneous melanoma spheroids, we used normal dermal fibroblasts from the sun-protected skin of a juvenile donor. We compared them to the fibroblasts from the sun-exposed photodamaged skin of an adult donor. Further, we analysed the spheroids by single-cell RNA sequencing. To validate transcriptional data, we also compared the immunohistochemical analysis of heterogeneous spheroids to melanoma biopsies. We have distinguished three functional clusters in primary human fibroblasts from melanoma spheroids. These clusters differed in the expression of (a) extracellular matrix-related genes, (b) pro-inflammatory factors, and (c) TGFβ signalling superfamily. We observed a broader deregulation of gene transcription in previously photodamaged cells. We have confirmed that pro-inflammatory cytokine IL-6 significantly enhances melanoma invasion to the extracellular matrix in our model. This supports the opinion that the aspects of ageing are essential for reliable melanoma 3D modelling in vitro.
    Permanent Link: http://hdl.handle.net/11104/0317376

     
     
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