Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcriptome profile of human pancreatic beta cell line (EndoC-M-NM-2H2 ) following knockdown of RFX6.

ABSTRACT: RFX6 is a key transcription factor for the development of mouse pancreas, however the functional roles of RFX6 in human beta cells are poorly explored. Thus transcriptome analysis was perfomed to determine the functional targets of RFX6 in human beta cells using the recently developed human beta cell-line EndoC-M-NM-2H2. Transcriptome profile of human beta cell line (EndoC-M-NM-2H2 cells) following siRNA induced knockdown of RFX6 is compared with siControl (siNT) treated EndoC-M-NM-2H2 cells.

ORGANISM(S): Homo sapiens

SUBMITTER: Raphael Scharfmann

PROVIDER: E-GEOD-59049 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

RFX6 regulates insulin secretion by modulating Ca2+ homeostasis in human β cells.

Chandra Vikash V Albagli-Curiel Olivier O Hastoy Benoit B Piccand Julie J Randriamampita Clotilde C Vaillant Emmanuel E Cavé Hélène H Busiah Kanetee K Froguel Philippe P Vaxillaire Martine M Rorsman Patrik P Polak Michel M Scharfmann Raphael R

Cell reports 20141211 6

Development and function of pancreatic β cells involve the regulated activity of specific transcription factors. RFX6 is a transcription factor essential for mouse β cell differentiation that is mutated in monogenic forms of neonatal diabetes. However, the expression and functional roles of RFX6 in human β cells, especially in pathophysiological conditions, are poorly explored. We demonstrate the presence of RFX6 in adult human pancreatic endocrine cells. Using the recently developed human β cel ...[more]

PMID: 25497100

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Project description:We previously established long-term 3D organoid culture systems for several murine tissues (intestine, stomach, pancreas and liver) as well as human intestine and pancreas. Here, we describe culture conditions to generate long-term 3D culture from human gastric stem cells. The technology can be applied to study the epithelial response to infection with Helicobacter pylori. Human gastric cultures can expand indefinitely in 3D Matrigel. Cultures can be generated from normal tissue, from single sorted stem cells, or from tumor tissue. Organoids maintain many characteristics of the respective tissue in terms of histology, marker expression and euploidy. Organoids from normal tissue express markers of four lineages of the stomach and self-organize in gland and pit-domains. They can be directed to specifically express either lineages of the gastric gland, or the gastric pit by addition of Nicotinamide and withdrawal of Wnt. While gastric pit lineages react marginally to bacterial infection, gastric gland lineages mount a strong inflammatory response. The gastric culture system provides a unique tool to study gastric pathologies. We generated 2 sets of experiments. The first set contains organoids in 4 conditions: (1) organoids in expansion condition ENRWFGNiTi ("gland-type organoids") from 3 donors, (2) organoids as in 1 but differentiated for 4 days in differentiation condition ENR_FGNiTi ("pit'type organoids"), (3) organoids as in 1 but infected with Helicobacter pylori strain P12 MOI 50 for 2 h, (4) organoids as in 2 but infected as in 3. All 4 conditions were tested on the same organoid line in parallel. This experiment was conducted independently with cultures from 3 different donors. The second set of experiments compares freshly isolated glands with organoids. Samples from 2 patients were analyzed. Each patient received a total gastrectomy. From each patient, glands from corpus region or pyloric antrum were isolated. From each isolation, one aliquod was stored for microarray analysis and one aliquod used to generate organoids. Organoids and glands were subsequently lysed and analyzed in parallel.

Dataset Information

Transcriptome profile of human pancreatic beta cell line (EndoC-M-NM-2H2 ) following knockdown of RFX6.

Publications

RFX6 regulates insulin secretion by modulating Ca2+ homeostasis in human β cells.

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