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The transcriptome expression differences between GDNF-treated and anti-NCAM1-treated EO771-tdT-LM2 cells under glucose-deprived media treatment

ABSTRACT: The transcriptome expression differences between GDNF-treated and anti-NCAM1-treated EO771-tdT-LM2 cells under glucose-deprived media treatment

PROVIDER: PRJNA1096326 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28559108_1.fastq.gz	Fastqsanger.gz
	SRR28559108_2.fastq.gz	Fastqsanger.gz
	SRR28559109_1.fastq.gz	Fastqsanger.gz
	SRR28559109_2.fastq.gz	Fastqsanger.gz
	SRR28559110_1.fastq.gz	Fastqsanger.gz

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The transcriptome expression differences between GDNF-treated and anti-NCAM1-treated EO771-tdT-LM2 cells under glucose-deprived media treatment

Project description:The molecular mechanisms that regulate breast cancer cell (BCC) metastasis and proliferation within the leptomeninges (LM) are poorly understood, limiting development of effective therapies. Here we show that BCCs in mice can invade the LM by abluminal migration along blood vessels that connect vertebral/calvarial bone marrow and meninges, bypassing the blood-brain barrier. This process is dependent on BCC engagement with vascular basement membrane laminin through expression of the neuronal pathfinding molecule integrin α6. Once in the LM, BCCs co-localize with perivascular meningeal macrophages and induce their expression of the pro-survival neurotrophin, GDNF. Intrathecal GDNF blockade, macrophage-specific GDNF ablation, or deletion of the GDNF receptor, NCAM, from BCCs, inhibit BC growth within LM. Here, we performed RNA-seq analysis of GDNF-treated and/or anti-NCAM1-treated EO771-tdT-LM2 cells under glucose-deprived media treatment.

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Project description:Adjacent stroma, including subepithelial fibroblasts, are believed to coordinate the differentiation process of epithelial cells, but the mechanisms are not well understood. Glial cell line-derived neurotrophic factor (GDNF) is expressed in the intestinal Pdgfra high subepithelial myofibroblasts (SEMFs), while the GDNF receptor RET is expressed in a subset of enteroendocrine cells (EECs), indicating regulatory crosstalk. In this experiment, single RET+ intestinal cells were sorted the gene expression profile was compared to non-RET cells. This provided knowledge about RET+ cell types. RET+ cells were upregulated with most of the EEC markers, indicating that RET+ cells are expressed in EECs. We also observed the differences in the gene expression profile of stromal-derived GDNF ligand on intestinal organoids. Organoids were derived from intestinal crypts of WT mice C57BL/6J and treated with 500ng/ml of Gdnf and Gfra1. GDNF-treated organoids induced the expression of EEC genes such as Pyy, Tac1, Tph1, and Cck, indicating enhanced differentiation of EC cell and L-I-N cell lineages. This highlights a stroma-epithelium crosstalk pathway regulating the differentiation of intestinal EEC subtypes.

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