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Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis [scRNA-seq]

ABSTRACT: Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis [scRNA-seq]

PROVIDER: PRJNA904756 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR22396971.fastq.gz	Fastqsanger.gz
	SRR22396972.fastq.gz	Fastqsanger.gz

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Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis

Project description:Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis

| PRJNA904761 | ENA

Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis [scRNA-seq]

Project description:Stem cell dynamics in the lung govern homeostasis, repair, and regeneration, yet there is still much unknown about the mechanisms of these processes. Furthermore, incongruencies between murine and human physiology limit the translation of some findings. In this work, we address these limitations by using a transgenic pig model to identify two populations of LGR5+ cells in the lung that are present in the human but that are absent from the mouse. Using RNA sequencing, 3D imaging, organoid models, and differentiation assays, we determine that in the fetal lung, epithelial LGR5 expression is transient in a subpopulation of developing lung bud tips. While epithelial LGR5 expression is absent from postnatal lung, it is reactivated in some organoids derived from basal airway cells. A separate population of LGR5+ cells is mesenchymal, surrounds developing and mature airways, is closely associated with nerve fibers, and acts as a multipotent progenitor cell capable of supporting the airway basal cell niche. These results point to two roles for LGR5 in orchestrating stem and progenitor cell dynamics, and provide a physiologically relevant model for further studies on the role of these populations in repair and regeneration.

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Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis [Bulk RNA-seq]

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Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis [Bulk RNA-seq]

Project description:Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis [Bulk RNA-seq]

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Transgenic porcine model reveals two roles for LGR5 during lung development and homeostasis

Project description:This SuperSeries is composed of the SubSeries listed below.

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LGR5 is a conserved marker of hair follicle stem cells across species

Project description:Using a transgenic pig expressing H2B-GFP under the control of the endogenous LGR5 promoter, we used fluorescence activated cell sorting to isolate LGR5-high and LGR5-negative epidermal cells to generate mRNA profiles of the hair follicle stem cell population, n=2 pigs. Bulk RNAseq samples were prepared from porcine cells, at least 500ng of RNA was extracted from sorted LGR5-GFP-high or LGR5-GFP-negative populations. RNAseq was performed externally by GENEWIZ; library preparation with poly(A) selection was performed followed by paired end 150bp sequencing on Illumina HiSeq.

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The porcine granulocyte proteome: expression changes in a transgenic pig model for diabetes

Project description:As transgenic INSC94Y-pigs develop a stable diabetic phenotype we were able to study the influence of elevated blood glucose levels on primary immune cells in vivo. We investigated the neutrophil proteome and compared data of transgenic animals to a control group. A total of 2371 proteins describing the whole neutrophil proteome were identified, including 396 proteins (17% of proteome) newly associated to expression in porcine neutrophils. Our studies provide novel information on the porcine granulocyte proteome and contribute to a better understanding of molecular mechanisms involved in altered immune cell function in diabetes. The data presented here is highly relevant for veterinary medicine and has translational quality for diabetes in human.

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Lgr4 maintains Lgr5+ crypt base columnar cell identity required for intestinal homeostasis and regeneration [RNA-Seq]

Project description:The intestinal epithelium is amongst the most rapidly self-renewing epithelia in mammals. Wnt signalling is known to be essential for intestinal homeostasis and regeneration. However, the underlying cellular hierarchy remains debated. Recent models have challenged dogma with a LGR4/FGBFBP1+ isthmus cell being upstream of the Lgr5⁺ crypt base columnar (CBC) cells. Importantly the precise functional roles of Lgr4 and Lgr5 not fully understood. Here we show that Lgr4 is broadly expressed within the intestinal epithelial cells (including in Lgr5+ CBCs) and is required directly for Lgr5+ CBCs, Paneth cells, long term crypt survival and regeneration. Lgr5 loss expands CBC markers and Paneth cells at the expense of Goblet cells. Loss of Lgr4 function can be rescued by loss of the E3 ubiquitin ligases ZNFR3/RNF43. Together, these findings establish LGR4 as the principal Wnt-potentiating Lgr receptor and an indispensable, regulator of Lgr5+ CBC cell identity in intestinal homeostasis and regeneration.

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Lgr4 maintains Lgr5+ crypt base columnar cell identity required for intestinal homeostasis and regeneration [scRNA-seq]

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