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Transcriptional profiling of healthy versus cystic fibrosis human endometrial organoids upon hormonal exposure

ABSTRACT: Transcriptional profiling of healthy versus cystic fibrosis human endometrial organoids upon hormonal exposure

PROVIDER: PRJEB80304 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR13695435.fastq.gz	Fastqsanger.gz
	ERR13695436.fastq.gz	Fastqsanger.gz
	ERR13695437.fastq.gz	Fastqsanger.gz
	ERR13695438.fastq.gz	Fastqsanger.gz
	ERR13695439.fastq.gz	Fastqsanger.gz

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Transcriptional profiling of healthy versus cystic fibrosis human endometrial organoids upon hormonal exposure

Project description:Female cystic fibrosis (CF) patients are confronted with fertility deficiency, experiencing difficulties in getting pregnant. Underlying reasons remain elusive. In particular, it has only poorly been studied whether the endometrium, the inner lining of the womb and crucial tissue for embryo implantation, plays a role. One main reason is the lack of reliable and tractable study models given the major technical and ethical hurdles to study events that occur hidden in the womb.To overcome this, we established an endometrium CF organoid model. We applied bulk RNA sequencing on these organoids derived form endometrium of women affected by CF and of healthy women. These endometrial organoids (EO) were exposed to hormones to mimic different phases of the menstrual cycle. The proliferative phase was mimicked by exposing the EO to estradiol (+E2), while the (mid-)secretory phase was mimicked by estradiol, progesterone, cyclic AMP and XAV-939 (Wnt inhibitor) (EPCX) exposure of EO. As a control, we sequenced healthy and CF EO not exposed to hormones (EOM).

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Gene expression of immortalized cystic fibrosis and non-cystic fibrosis airway epithelial cells

Project description:Our laboratory has held a long interest in the glycosylation changes seen on the surface of airway epithelia of patients with the disease cystic fibrosis (CF). Experiments from our laboratory have detailed a CF glycosylation phenotype of increased Fuca1,3/4 and decreased Fuca1,2 and sialic acid on the surfaces of immortalized and primary CF cells compared to non-CF cells. Further, we have shown that gene transfer and subsequent expression of a wild type CF plasmid in CF airway cells results in correction or reversal of this glycosylation phenotype. We hypothesize that the changes in glycosylation seen in CF cells are key in the pathophysiology of the cystic fibrosis airway disease. For example, it has been shown that Pseudomonas aeruginosa, a bacterium that has a predilection for colonizing CF airways, adheres to asialylated glycolipids and glycoconjugates with terminal Fuca1,3/4. One focus of our laboratory is to elucidate the etiology of the glycosylation changes seen in CF cells and the mechanism by which these changes are reversed by wild type CFTR gene transfer. We propose to study the gene expression of immortalized and primary CF and non-CF airway epithelial cells: 1. CF/T43 vs. BEAS-2B cells. These are two widely used immortalized airway cell lines that we have used extensively in the past. 2. C38 cells; C38 cells are IB3 cells expressing wtCFTR. The experimental focus is to elucidate the etiology of the glycosylation changes seen in Cystic Fibrosis (CF) cells and the mechanism by which these changes are reversed by wild type CFTR gene transfer. To do so, the gene expression of immortalized and primary CF and non-CF airway epithelial cells were compared and studied. Cell lines used were CF/T43 and BEAS-2B, both widely used immortalized airway cell lines. Other cell lines studied included C38 cell lines (clonal derivatives of IB3 cells expressing wtCFTR).

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