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MRNA sequencing of stem cell-derived beta cells before and after transplantation in mice

ABSTRACT: mRNA sequencing of stem cell-derived beta cells before and after transplantation in mice

PROVIDER: PRJNA613926 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11365000_1.fastq.gz	Fastqsanger.gz
	SRR11365000_2.fastq.gz	Fastqsanger.gz
	SRR11365001_1.fastq.gz	Fastqsanger.gz
	SRR11365001_2.fastq.gz	Fastqsanger.gz
	SRR11365002_1.fastq.gz	Fastqsanger.gz

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mRNA sequencing of stem cell-derived beta cells before and after transplantation in mice

Project description:A drastic transition at birth, from constant maternal nutrient supply in utero to intermittent postnatal feeding, requires changes in the metabolic system. Despite their central role in metabolic homeostasis, little is known about how pancreatic beta cells adjust to the new nutritional program. Our experiments show that after birth beta cells shift from amino acid- to glucose-stimulated insulin secretion that correlates with the nutritional environment of the animal. This metabolic adaptation is mediated by a transition in nutrient sensitivity of the mTORC1 pathway , which leads toresults in intermittent mTORC1 signaling activity. Disrupting nutrient sensitivity of mTORC1 in mature beta cells affects insulin secretion and reverts them to an immature functional state. Finally, manipulating nutrient sensitivity of mTORC1 in stem cell-derived beta cells in vitro strongly enhances their glucose-responsive insulin secretion. These results reveal a mechanism by which nutrient sensing regulates beta cell function, thereby enabling a metabolic adaptation for the newborn.

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Project description:Purpose: Pancreatic islet transplantation is an effective cell therapy for type 1 diabetes (T1D), but its clinical application is limited by the shortage of donor pancreata. Among the potential alternatives, the differentiation of human embryonic stem cells (hESc) into insulin-producing β-cells has taken an early lead. However, while the proportion of β-cells obtained through current methods is relatively high, a significant percentage of undefined non-endocrine cell types are still generated. Most importantly, there is the potential for carry-over of non-differentiated cell types that may produce teratomas upon transplantation. In order to address these issues, we sought to modify hESc so that their differentiated progeny could be selectively devoid of tumorigenic cells and enriched for cells of the desired phenotype (in this case, pancreatic β-cells). Methods: mRNA profiles of λH1-hESC derived β-like cells were developed by mRNA sequencing, in triplicate, using Illumina HiSeq PE Cluster Kit v4 and Illumina HiSeq Flow Cell v4 with 50 nt paired end reads plus dual index reads using the Illumina HiSeq SBS kit v4. Sequence reads that passed quality filters were analyzed at the transcript isoform level following alignment using TopHat v2.1.0 followed by exon and gene level counting using Bioconductor easyRNASeq v 2.4.7. Conclusions: Our study demonstrates that λH1-hESC derived β-like cells have a transcriptional expression profile similar to ESC derived β-like cells.

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Single-Cell Transcriptome Profiling Reveals β Cell Maturation in Stem Cell-Derived Islets after Transplantation

Project description:SC-β cells and native β cells from cadaveric islets have different transcriptome profiles. In this study, we demonstrate that SC-β cells can acquire mature gene signatures via transplantation in mice for 6 months. MAFA, a marker for mature β cells, is expressed and resulted in significant improvements in insulin release and glucose regulation in diabetic mice.

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