A Coalgebraic View of Infinite Trees and Iteration

AbstractThe algebra of infinite trees is, as proved by C. Elgot, completely iterative, i.e., all ideal recursive equations are uniquely solvable. This is proved here to be a general coalgebraic phenomenon: let H be an endofunctor such that for every object X a final coalgebra, TX, of H(_) + X exists. Then TX is an object-part of a monad which is completely iterative. Moreover, a similar contruction of a “completely iterative monoid” is possible in every monoidal category satisfying mild side conditions

Similar ex vivo expansion and post-irradiation regenerative potential of juvenile and aged salivary gland stem cells

AbstractBackground and purposeSalivary gland dysfunction is a major side effect of radiotherapy for head and neck cancer patients, which in the future might be salvaged by autologous adult salivary gland stem cell (SGSC) therapy. Since frail elderly patients may have decreased activity of SGSCs, we aimed to characterize the potential of aged SGSC-population in a murine model.Materials and methodsSalivary glands and salisphere-derived cells from young and old mice were tested for CD24 and CD29 stem cell marker expression using FACS. Moreover, in vitro expansion capability and in vivo regeneration potential upon post-irradiation transplantation of young and aged SGSCs were measured.ResultsAn increase in CD24hi/CD29hi putative stem cells was detected in aged salivary glands albeit with a decrease in functional ability to form salispheres. However, the salispheres formed from aged mice salivary glands expressed CD24hi/CD29hi to the same extent as the ones from young mice. Moreover, following exposure to adequate growth conditions old and young SGSCs exhibited similar in vitro expansion- and in vivo regeneration potential.ConclusionsAged SGSCs although reduced in number are in vitro indistinguishable from young SGSCs and could potentially be used to ameliorate age- or treatment related salivary gland dysfunction

Role of quiescent cells in the homeostatic maintenance of the adult submandibular salivary gland

Stem/progenitor cells are required for maintenance of salivary gland (SG) function and serve as untapped reservoirs to create functional cells. Despite recent advancements in the identification of stem/progenitor pools, in the submandibular gland (SMG), a knowledge gap remains. Furthermore, the contribution to adult SMG homeostasis of stem/progenitor cells originating from embryonic development is unclear. Here, we employ an H2B-GFP embryonic and adult pulse-and-chase system to characterize potential SMG stem/progenitor cells (SGSCs) based on quiescence at different stages. Phenotypical profiling of quiescent cells in the SMG revealed that label-retaining cells (LRCs) of embryonic or adult origin co-localized with CK8+ ductal or vimentin+ mesenchymal, but not with CK5+ or CK14+ stem/progenitor cells. These SMG LRCs failed to self-renew in vitro while non-label retaining cells displayed differentiation and long-term expansion potential as organoids. Collectively, our data suggest that an active cycling population of cells is responsible for SMG homeostasis with organoid forming potential

Transcriptional and epigenomic profiling identifies YAP signaling as a key regulator of intestinal epithelium maturation

During intestinal organogenesis, equipotent epithelial progenitors mature into phenotypically distinct stem cells that are responsible for lifelong maintenance of the tissue. While the morphological changes associated with the transition are well characterized, the molecular mechanisms underpinning the maturation process are not fully understood. Here, we leverage intestinal organoid cultures to profile transcriptional, chromatin accessibility, DNA methylation, and three-dimensional (3D) chromatin conformation landscapes in fetal and adult epithelial cells. We observed prominent differences in gene expression and enhancer activity, which are accompanied by local changes in 3D organization, DNA accessibility, and methylation between the two cellular states. Using integrative analyses, we identified sustained Yes-Associated Protein (YAP) transcriptional activity as a major gatekeeper of the immature fetal state. We found the YAP-associated transcriptional network to be regulated at various levels of chromatin organization and likely to be coordinated by changes in extracellular matrix composition. Together, our work highlights the value of unbiased profiling of regulatory landscapes for the identification of key mechanisms underlying tissue maturation

YAP/TAZ-Dependent Reprogramming of Colonic Epithelium Links ECM Remodeling to Tissue Regeneration.

Tissue regeneration requires dynamic cellular adaptation to the wound environment. It is currently unclear how this is orchestrated at the cellular level and how cell fate is affected by severe tissue damage. Here we dissect cell fate transitions during colonic regeneration in a mouse dextran sulfate sodium (DSS) colitis model, and we demonstrate that the epithelium is transiently reprogrammed into a primitive state. This is characterized by de novo expression of fetal markers as well as suppression of markers for adult stem and differentiated cells. The fate change is orchestrated by remodeling the extracellular matrix (ECM), increased FAK/Src signaling, and ultimately YAP/TAZ activation. In a defined cell culture system recapitulating the extracellular matrix remodeling observed in vivo, we show that a collagen 3D matrix supplemented with Wnt ligands is sufficient to sustain endogenous YAP/TAZ and induce conversion of cell fate. This provides a simple model for tissue regeneration, implicating cellular reprogramming as an essential element.This work was supported by Worldwide Cancer Research (13-1216 to KBJ), Lundbeck Foundation (R105-A9755 to KBJ), the Danish Cancer Society (R56-A2907 and R124-A7724 to KBJ), the Carlsberg Foundation (to KBJ), EMBO Young Investigator programme (to KBJ), AIRC Special Program Molecular Clinical Oncology ‘‘5 per mille’’ (to SP), an AIRC PI-Grant (to SP), Epigenetics Flagship projects (CNR-Miur grants. to SP), the DFF mobilix programme (to SY), Marie Curie fellowship programme (SY and JG), Foundation of Aase and Ejnar Danielsen (OHN), Axel Muusfeldts Foundation (OHN), The Ragnar Söderberg Foundation (CDM). This project has received funding from the European Union’s Horizon 2020 research and innovation programme (grant agreements STEMHEALTH ERCCoG682665 and INTENS 668294 to KBJ and DENOVOSTEM No. 670126 to SP)

YAP/TAZ-Dependent Reprogramming of Colonic Epithelium Links ECM Remodeling to Tissue Regeneration

Tissue regeneration requires dynamic cellular adaptation to the wound environment. It is currently unclear how this is orchestrated at the cellular level and how cell fate is affected by severe tissue damage. Here we dissect cell fate transitions during colonic regeneration in a mouse dextran sulfate sodium (DSS) colitis model, and we demonstrate that the epithelium is transiently reprogrammed into a primitive state. This is characterized by de novo expression of fetal markers as well as suppression of markers for adult stem and differentiated cells. The fate change is orchestrated by remodeling the extracellular matrix (ECM), increased FAK/Src signaling, and ultimately YAP/TAZ activation. In a defined cell culture system recapitulating the extracellular matrix remodeling observed in vivo, we show that a collagen 3D matrix supplemented with Wnt ligands is sufficient to sustain endogenous YAP/TAZ and induce conversion of cell fate. This provides a simple model for tissue regeneration, implicating cellular reprogramming as an essential element

Nocodazole Treatment Decreases Expression of Pluripotency Markers Nanog and Oct4 in Human Embryonic Stem Cells

Nocodazole is a known destabiliser of microtubule dynamics and arrests cell-cycle at the G2/M phase. In the context of the human embryonic stem cell (hESC) it is important to understand how this arrest influences the pluripotency of cells. Here we report for the first time the changes in the expression of transcription markers Nanog and Oct4 as well as SSEA-3 and SSEA-4 in human embryonic cells after their treatment with nocodazole. Multivariate permeabilised-cell flow cytometry was applied for characterising the expression of Nanog and Oct4 during different cell cycle phases. Among untreated hESC we detected Nanog-expressing cells, which also expressed Oct4, SSEA-3 and SSEA-4. We also found another population expressing SSEA-4, but without Nanog, Oct4 and SSEA-3 expression. Nocodazole treatment resulted in a decrease of cell population positive for all four markers Nanog, Oct4, SSEA-3, SSEA-4. Nocodazole-mediated cell-cycle arrest was accompanied by higher rate of apoptosis and upregulation of p53. Twenty-four hours after the release from nocodazole block, the cell cycle of hESC normalised, but no increase in the expression of transcription markers Nanog and Oct4 was detected. In addition, the presence of ROCK-2 inhibitor Y-27632 in the medium had no effect on increasing the expression of pluripotency markers Nanog and Oct4 or decreasing apoptosis or the level of p53. The expression of SSEA-3 and SSEA-4 increased in Nanog-positive cells after wash-out of nocodazole in the presence and in the absence of Y-27632. Our data show that in hESC nocodazole reversible blocks cell cycle, which is accompanied by irreversible loss of expression of pluripotency markers Nanog and Oct4

Potential of salivary gland stem cells in regenerative medicine

Weefselspecifieke volwassen stamcellen hebben de aandacht getrokken van de biomedische gemeenschap doordat ze in staat zijn om alle celtypes van een orgaan te vormen waardoor ze bruikbaar zouden kunnen zijn voor stamceltherapie. De hoop is dat deze cellen beschadigd weefsel kunnen herstellen en wellicht zelfs voor volledige vervanging van organen kunnen zorgen. Zo zouden bijvoorbeeld volwassen stamcellen die in staat zijn tot vorming van speekselklierweefsel gebruikt kunnen worden om patiënten die lijden aan het droge mond syndroom (Xerostomie) te behandelen. Xerostomie houdt in dat te weinig speeksel wordt geproduceerd en is een veel voorkomende bijwerking als gevolg van radiotherapie bij hoofd- en hals-tumoren, Sjögren-syndroom, diabetes, ouderdom en het gebruik van sommige medicijnen. Het is echter van het grootste belang dat de nog ontbrekende informatie over de identiteit, lokalisatie, en de kritieke moleculaire signalen die nodig zijn voor het handhaven en kweken in een laboratorium van volwassen speekselklier stamcellen tot in detail onderzocht worden. Het onderzoek beschreven in dit proefschrift draagt hieraan bij en verbetert de huidige kennis over de identiteit van de speekselklierstamcel en de moleculaire signaal routes betrokken bij speekselklier regeneratie. Verder wordt de optimalisatie van een kweekmethode voor volwassen speekselklier stamcellen van muis en mens beschreven. Deze kennis wordt toegepast om een beter begrip van de speekselklier stamcelbiologie tijdens weefsel homeostasis, regeneratie en veroudering te krijgen. De in dit proefschrift gepresenteerde gegevens benadrukken het veelbelovende therapeutische potentieel van speekselklier stamcellen in regeneratieve geneeskunde