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Transcriptomics informs design of a planar human enterocyte culture system that reveals metformin enhances fatty acid flux

ABSTRACT: Transcriptomics informs design of a planar human enterocyte culture system that reveals metformin enhances fatty acid flux

PROVIDER: PRJNA774642 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16575100.fastq.gz	Fastqsanger.gz
	SRR16575101.fastq.gz	Fastqsanger.gz
	SRR16575102.fastq.gz	Fastqsanger.gz
	SRR16575103.fastq.gz	Fastqsanger.gz
	SRR16575104.fastq.gz	Fastqsanger.gz

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Transcriptomics informs design of a planar human enterocyte culture system that reveals metformin enhances fatty acid flux

Project description:Background & Aims: Absorption, metabolism, and secretion of lipids occurs in the small intestinal epithelium. Caco-2 and organoids have been used to study these processes but are limited in physiological relevance or preclude simultaneous apical and basal access. Here, we develop a dynamic and flexible planar absorptive enterocyte (AE) monolayer system, investigate how fatty acid oxidation (FAO) regulates FA mobilization mechanisms and probe a role for metformin in FA mobilization. Methods: Single-cell RNA-sequencing (scRNAseq) was performed on primary human jejunum. Transcriptomic signatures and trajectory analysis defined in vivo AE maturational signatures, which informed culture methods to differentiate ISCs and mimic in vivo AEs. The system was scaled for high-throughput drug screening and FAO was pharmacologically modulated. BODIPY (B)-labelled fatty acids (FAs) were used to easily and sensitively evaluate FA handling via fluorescence, and thin layer chromatography (TLC). Results: scRNAseq shows increasing expression of lipid-handling genes as AEs mature. In vitro, ISCs differentiate into AEs that mimic the in vivo maturational program, exhibit strong barrier function and express FA-handling genes. FA synthase inhibitor, C75, increased apical to basolateral mobilization of B-C16. Short-chain FA (B-C5) was unaffected and diffused freely through AEs. FAO inhibitor, etomoxir, decreased apical to basolateral mobilization of medium and long chain FAs (B-C12, B-C16) whereas anti-diabetic drug, metformin, augmented this mobilization. Conclusions: Primary human ISCs in culture undergo programmed maturation in vitro to generate a sensitive culture system of absorptive epithelium to investigate FA-handling. Modulating FAO impacts mobilization of FAs across absorptive epithelium and FAO enhanced by metformin increases basolateral mobilization pointing to an intestine-specific role.

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Project description:This SuperSeries is composed of the following subset Series: GSE30558: Bisulphite-sequencing of chromatin immunoprecipitated DNA (BisChiP-seq) directly informs methylation status GSE34340: Bisulphite sequencing of native LNCaP and PrEC DNA [methylation array] Refer to individual Series

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Regulation of Drosophila intestinal stem cell proliferation by enterocyte mitochondrial pyruvate metabolism

Project description:Purpose: The goals of this study was to compare differentially expressed transcripts when MPC expression is supressed in the adult intestines or enterocyte. The NCBI gene ID for dMPC1 is 42268.

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Project description:The intestinal epithelium consistently exposes to various injuries and exhibits remarkable regenerative capacity. Although small intestinal regeneration has been extensively investigated, the mechanisms governing colonic regeneration remain poorly understood. Here, we identify irradiation-induced regenerative cells (IRECs), a previously unrecognized colonic enterocyte population, emerges following irradiation injury. IRECs, marked by the gene YuanShangCao (Ysc), undergo dedifferentiation and transition into a stem-like state, contributing to epithelial regeneration. IREC ablation severely impairs colonic repair. Mechanistically, YSC promotes colonic regeneration by preventing the lysosomal degradation of YAP. Furthermore, we demonstrate a critical role of p53 in colonic regeneration by inducing Ysc expression. Together, our findings define IRECs as an irradiation injury-induced enterocyte population that dedifferentiate into stem cells in colon epithelial repair and establish the p53-YSC-YAP axis plays a key role in this process, deepening the understanding of colonic diseases.

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