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Endothelial cells synergize with fibroblasts to form a niche for engineering alveolar epithelium

ABSTRACT: Endothelial cells synergize with fibroblasts to form a niche for engineering alveolar epithelium

PROVIDER: PRJNA738933 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14855048.fastq.gz	Fastqsanger.gz
	SRR14855049.fastq.gz	Fastqsanger.gz
	SRR14855050.fastq.gz	Fastqsanger.gz
	SRR14855051.fastq.gz	Fastqsanger.gz
	SRR14855052.fastq.gz	Fastqsanger.gz

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Endothelial cells synergize with fibroblasts to form a niche for engineering alveolar epithelium

Project description:We perform single-cell RNA sequencing of whole engineered lungs generated either by alveolar epithelial type 2 cell (AEC2)/fibroblast (FB) co-culture or by AEC2/FB/endothelial cell (EC) tri-culture, to investigate AEC2 and FB phenotype in the presence and absence of endothelial cells. Primary postnatal day 7 (P7) AEC2s, P7 lung FBs, and primary rat lung microvascular ECs (from 4-6 week-old rats) used for engineered cultures, as well as native P7 whole rat lung, were sequenced for comparison. We find that native-like AEC2 phenotype, alveolar structure, and cellular signaling patterns are better recapitulated via epithelial-mesenchymal-endothelial tri-culture rather than by a more simplified system. FBs cultured in the presence of endothelial cells demonstrate reduced expression of contractile markers and upregulate features of an AEC2 niche-supportive signature.

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Sympathetic nerves and arrector pili muscles form a dual component niche to regulate hair follicle stem cells

Project description:Piloerection (goosebump) requires concerted actions of the hair follicle, the arrector pili muscle (APM), and the sympathetic nerve, providing a model to study interactions across epithelium, mesenchyme, and nerves. Here, we show that APMs and sympathetic nerves form a dual component niche to modulate hair follicle stem cell (HFSC) activity. Sympathetic nerves form synapse-like structures with HFSCs and regulate HFSCs through norepinephrine, whereas APMs maintain sympathetic innervation to HFSCs. Without norepinephrine signaling, HFSCs enter a deep quiescence state by down-regulating cell cycle machinery and mitochondria metabolism, while up-regulating quiescence regulators Lhx2, Foxp1, and Fgf18. During development, HFSC progeny secrets Sonic Hedgehog (SHH) to direct the formation of this APM-sympathetic nerve niche, which in turn controls hair follicle regeneration in adults. Our results reveal a reciprocal interdependence between a regenerative tissue and its niche at different stages, and illustrate that nerves can modulate stem cell quiescence through synapses and neurotransmitters.

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Project description:Engineering a Niche Supporting Hematopoietic Stem Cell Development Using Integrated Single Cell Transcriptomics

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Engineering a Niche Supporting Hematopoietic Stem Cell Development Using Integrated Single Cell Transcriptomics

Project description:Hematopoietic stem cells (HSC) develop from hemogenic endothelium (HE) within embryonic arterial vessels such as the aorta of the aorta-gonad-mesonephros region (AGM). To identify the signals responsible for HSC formation, we used single cell RNA-sequencing to simultaneously analyze the transcriptional profiles of AGM-derived cells transitioning from HE to HSC, and AGM-derived endothelial cells which provide signals sufficient to support HSC maturation and self-renewal. Pseudotemporal ordering revealed dynamics of gene expression during the HE to HSC transition, identifying surface receptors specifically expressed on developing HSC. Transcriptional profiles of niche endothelial cells enabled identification of corresponding ligands, including those signaling to Notch receptors, VLA-4 integrin, and CXCR4, which, when integrated in an engineered platform, were sufficient to support the generation of engrafting HSC. These studies provide a transcriptional map of the signaling interactions necessary for the development of HSC and advance the goal of engineering HSC for therapeutic applications

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Sentinel p16INK4a+ cells in the basement membrane form a reparative niche in the lung

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Synergistic, context-dependent and hierarchical functions of hematopoietic progenitor niche components.

Project description:Specialized niche environments specify and maintain stem and progenitor cells, but little is known about the identities and functional interactions of niche components in vivo. Here, we describe a modular system for the generation of artificial hematopoietic niches in the mouse embryo. A circumscribed tissue that lacks niche function but is physiologically accessible for hematopoietic progenitor cells is functionalized by individual and combinatorial expression of four factors, the chemokines Ccl25 and Cxcl12, the cytokine Scf and the Notch ligand DLL4. The distinct phenotypes and variable numbers of hematopoietic cells in the resulting niches reveal synergistic, context-dependent and hierarchical interactions among niche effector molecules. The surprisingly simple rules determining niche outcomes enable the in vivo engineering of artificial niches conducive to the presence of distinct myeloid or T or B lymphoid lineage precursors. The dataset comprises 24 samples divided into eight sample groups each representing a different lymphoid progenitor cell type isolated from wild-type (+/-) or transgenic (-/-) thymic niches. -/-, Foxn1-deficient genotype; +/-, Foxn1 heterozygous phenotype; DP, CD4/CD8 double-poisztive thymocytes; DN3, CD4/CD8-negative stage 3 thymocytes; SP4, CD4 single-positive thymocytes; SP8, CD8 single-positive thymocytes; B IgM-, IgM surface negative B cells; B IgM+, IgM surface positive B cells; B IgM- -/-, IgM surface negative B cells from Foxn1-deficient genotype.

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Lgr5-expressing secretory cells form a Wnt inhibitory niche in cartilage critical for chondrocyte identity

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