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Epigenetic Erasing and Pancreatic Differentiation of Dermal Fibroblasts into Insulin-Producing Cells are Boosted by the Use of Low-Stiffness Substrate

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Abstract

Several studies have demonstrated the possibility to revert differentiation process, reactivating hypermethylated genes and facilitating cell transition to a different lineage. Beside the epigenetic mechanisms driving cell conversion processes, growing evidences highlight the importance of mechanical forces in supporting cell plasticity and boosting differentiation. Here, we describe epigenetic erasing and conversion of dermal fibroblasts into insulin-producing cells (EpiCC), and demonstrate that the use of a low-stiffness substrate positively influences these processes. Our results show a higher expression of pluripotency genes and a significant bigger decrease of DNA methylation levels in 5-azacytidine (5-aza-CR) treated cells plated on soft matrix, compared to those cultured on plastic dishes. Furthermore, the use of low-stiffness also induces a significant increased up-regulation of ten-eleven translocation 2 (Tet2) and histone acetyltransferase 1 (Hat1) genes, and more decreased histone deacetylase enzyme1 (Hdac1) transcription levels. The soft substrate also encourages morphological changes, actin cytoskeleton re-organization, and the activation of the Hippo signaling pathway, leading to yes-associated protein (YAP) phosphorylation and its cytoplasmic translocation. Altogether, this results in increased epigenetic conversion efficiency and in EpiCC acquisition of a mono-hormonal phenotype. Our findings indicate that mechano-transduction related responsed influence cell plasticity induced by 5-aza-CR and improve fibroblast differentiation toward the pancreatic lineage.

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Acknowledgements

This work was funded by Carraresi Foundation, by European Foundation for the Study of Diabetes (EFSD) and by Ricerca Corrente at Centro Cardiologico Monzino, IRCCS. The Laboratory of Biomedical Embryology is member of the COST Action CA16119 In vitro 3-D total cell guidance and fitness (CellFit), the COST Action BM1308 Sharing advances on large animal models (SALAAM) and the COST Action CM1406 Epigenetic Chemical Biology (EPICHEM). The Authors are indebted with Drs Loredana Casalis and Denis Scaini at ELETTRA-Sincrotrone for helping with assessment of the substrates compliance with AFM-assisted nano-indentation methods.

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Authors and Affiliations

  1. Laboratory of Biomedical Embryology, Center for Stem Cell Research, Università degli Studi di Milano, Via Celoria 10, Milan, 20133, Italy

    Georgia Pennarossa, Elena F. M. Manzoni, Fulvio Gandolfi & Tiziana A. L. Brevini

  2. Unità di Ingegneria Tissutale Cardiovascolare, Centro Cardiologico Monzino-IRCCS, Milan, 20138, Italy

    Rosaria Santoro & Maurizio Pesce

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  1. Georgia Pennarossa
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  2. Rosaria Santoro
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  3. Elena F. M. Manzoni
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  4. Maurizio Pesce
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  5. Fulvio Gandolfi
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  6. Tiziana A. L. Brevini
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Correspondence to Tiziana A. L. Brevini.

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Pennarossa, G., Santoro, R., Manzoni, E.F.M. et al. Epigenetic Erasing and Pancreatic Differentiation of Dermal Fibroblasts into Insulin-Producing Cells are Boosted by the Use of Low-Stiffness Substrate. Stem Cell Rev and Rep 14, 398–411 (2018). https://doi.org/10.1007/s12015-017-9799-0

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