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Plasticity of adult human pancreatic duct cells by neurogenin3-mediated reprogramming.

Swales N, Martens GA, Bonné S, Heremans Y, Borup R, Van de Casteele M, Ling Z, Pipeleers D, Ravassard P, Nielsen F, Ferrer J, Heimberg H - PLoS ONE (2012)

Bottom Line: This neuro-endocrine shift however, is incomplete with less than 10% of full duct-to-endocrine reprogramming achieved.Transduction of exogenous Ngn3 activates endogenous Ngn3 suggesting auto-activation of this gene.Furthermore, pancreatic endocrine reprogramming of human duct cells can be moderately enhanced by inhibition of Delta-Notch signaling as well as by co-expressing the transcription factor Myt1, but not MafA and Pdx1.

View Article: PubMed Central - PubMed

Affiliation: Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium.

ABSTRACT

Aims/hypothesis: Duct cells isolated from adult human pancreas can be reprogrammed to express islet beta cell genes by adenoviral transduction of the developmental transcription factor neurogenin3 (Ngn3). In this study we aimed to fully characterize the extent of this reprogramming and intended to improve it.

Methods: The extent of the Ngn3-mediated duct-to-endocrine cell reprogramming was measured employing genome wide mRNA profiling. By modulation of the Delta-Notch signaling or addition of pancreatic endocrine transcription factors Myt1, MafA and Pdx1 we intended to improve the reprogramming.

Results: Ngn3 stimulates duct cells to express a focused set of genes that are characteristic for islet endocrine cells and/or neural tissues. This neuro-endocrine shift however, is incomplete with less than 10% of full duct-to-endocrine reprogramming achieved. Transduction of exogenous Ngn3 activates endogenous Ngn3 suggesting auto-activation of this gene. Furthermore, pancreatic endocrine reprogramming of human duct cells can be moderately enhanced by inhibition of Delta-Notch signaling as well as by co-expressing the transcription factor Myt1, but not MafA and Pdx1.

Conclusions/interpretation: The results provide further insight into the plasticity of adult human duct cells and suggest measurable routes to enhance Ngn3-mediated in vitro reprogramming protocols for regenerative beta cell therapy in diabetes.

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Related in: MedlinePlus

Ngn3 induces endocrine cell-enriched and neural-enriched genes in ducts.We analyzed all transcripts that are Ngn3-activated in suspension cultured duct cells (n = 140 on HG133A) in a public dataset of more than 50 human tissues and cell types (each n = 2), supplemented by our endocrine cell hybridizations (n = 3). The cluster graph thus shows the relative tissue-selectivity of the Ngn3-induced transcripts. Approximately 25% of Ngn3-regulated transcripts are selectively enriched in endocrine cells/pancreatic islets (cluster A). Cluster B genes are enriched in the central nervous system but generally not in pancreatic endocrine cells. Cluster C exhibit a neuro-endocrine expression pattern, with comparable expression in central nervous system and endocrine cells. An annotated version of Figure 1 with full gene names is included as Figure S2.
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pone-0037055-g001: Ngn3 induces endocrine cell-enriched and neural-enriched genes in ducts.We analyzed all transcripts that are Ngn3-activated in suspension cultured duct cells (n = 140 on HG133A) in a public dataset of more than 50 human tissues and cell types (each n = 2), supplemented by our endocrine cell hybridizations (n = 3). The cluster graph thus shows the relative tissue-selectivity of the Ngn3-induced transcripts. Approximately 25% of Ngn3-regulated transcripts are selectively enriched in endocrine cells/pancreatic islets (cluster A). Cluster B genes are enriched in the central nervous system but generally not in pancreatic endocrine cells. Cluster C exhibit a neuro-endocrine expression pattern, with comparable expression in central nervous system and endocrine cells. An annotated version of Figure 1 with full gene names is included as Figure S2.

Mentions: 140 transcripts on HG133A array transcripts are at least 1.5-fold (LB) activated by AdGFP-Ngn3 in human duct cells. Ngn3-activated genes are statistically enriched (P<0.001) in gene ontologies of development, neurogenesis, synaptic transmission, regulated hormone secretion and nuclear regulators; genes belonging to these clusters are shown. From left to right, columns indicate fold transcript accumulation in AdGFP-Ngn3 versus AdGFP control cells at 3 (D3) or 14 (D14) days post transduction. Two columns on the right refer to the analysis of tissue tropism in Figure 1: endocrine cell abundance indicates relative mRNA abundance in human endocrine cells as compared to the other tissues profiled in Figure 1. Last column indicates in which cluster (A, B, C) of Figure 1 the gene is situated.


Plasticity of adult human pancreatic duct cells by neurogenin3-mediated reprogramming.

Swales N, Martens GA, Bonné S, Heremans Y, Borup R, Van de Casteele M, Ling Z, Pipeleers D, Ravassard P, Nielsen F, Ferrer J, Heimberg H - PLoS ONE (2012)

Ngn3 induces endocrine cell-enriched and neural-enriched genes in ducts.We analyzed all transcripts that are Ngn3-activated in suspension cultured duct cells (n = 140 on HG133A) in a public dataset of more than 50 human tissues and cell types (each n = 2), supplemented by our endocrine cell hybridizations (n = 3). The cluster graph thus shows the relative tissue-selectivity of the Ngn3-induced transcripts. Approximately 25% of Ngn3-regulated transcripts are selectively enriched in endocrine cells/pancreatic islets (cluster A). Cluster B genes are enriched in the central nervous system but generally not in pancreatic endocrine cells. Cluster C exhibit a neuro-endocrine expression pattern, with comparable expression in central nervous system and endocrine cells. An annotated version of Figure 1 with full gene names is included as Figure S2.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351393&req=5

pone-0037055-g001: Ngn3 induces endocrine cell-enriched and neural-enriched genes in ducts.We analyzed all transcripts that are Ngn3-activated in suspension cultured duct cells (n = 140 on HG133A) in a public dataset of more than 50 human tissues and cell types (each n = 2), supplemented by our endocrine cell hybridizations (n = 3). The cluster graph thus shows the relative tissue-selectivity of the Ngn3-induced transcripts. Approximately 25% of Ngn3-regulated transcripts are selectively enriched in endocrine cells/pancreatic islets (cluster A). Cluster B genes are enriched in the central nervous system but generally not in pancreatic endocrine cells. Cluster C exhibit a neuro-endocrine expression pattern, with comparable expression in central nervous system and endocrine cells. An annotated version of Figure 1 with full gene names is included as Figure S2.
Mentions: 140 transcripts on HG133A array transcripts are at least 1.5-fold (LB) activated by AdGFP-Ngn3 in human duct cells. Ngn3-activated genes are statistically enriched (P<0.001) in gene ontologies of development, neurogenesis, synaptic transmission, regulated hormone secretion and nuclear regulators; genes belonging to these clusters are shown. From left to right, columns indicate fold transcript accumulation in AdGFP-Ngn3 versus AdGFP control cells at 3 (D3) or 14 (D14) days post transduction. Two columns on the right refer to the analysis of tissue tropism in Figure 1: endocrine cell abundance indicates relative mRNA abundance in human endocrine cells as compared to the other tissues profiled in Figure 1. Last column indicates in which cluster (A, B, C) of Figure 1 the gene is situated.

Bottom Line: This neuro-endocrine shift however, is incomplete with less than 10% of full duct-to-endocrine reprogramming achieved.Transduction of exogenous Ngn3 activates endogenous Ngn3 suggesting auto-activation of this gene.Furthermore, pancreatic endocrine reprogramming of human duct cells can be moderately enhanced by inhibition of Delta-Notch signaling as well as by co-expressing the transcription factor Myt1, but not MafA and Pdx1.

View Article: PubMed Central - PubMed

Affiliation: Diabetes Research Center, Vrije Universiteit Brussel, Brussels, Belgium.

ABSTRACT

Aims/hypothesis: Duct cells isolated from adult human pancreas can be reprogrammed to express islet beta cell genes by adenoviral transduction of the developmental transcription factor neurogenin3 (Ngn3). In this study we aimed to fully characterize the extent of this reprogramming and intended to improve it.

Methods: The extent of the Ngn3-mediated duct-to-endocrine cell reprogramming was measured employing genome wide mRNA profiling. By modulation of the Delta-Notch signaling or addition of pancreatic endocrine transcription factors Myt1, MafA and Pdx1 we intended to improve the reprogramming.

Results: Ngn3 stimulates duct cells to express a focused set of genes that are characteristic for islet endocrine cells and/or neural tissues. This neuro-endocrine shift however, is incomplete with less than 10% of full duct-to-endocrine reprogramming achieved. Transduction of exogenous Ngn3 activates endogenous Ngn3 suggesting auto-activation of this gene. Furthermore, pancreatic endocrine reprogramming of human duct cells can be moderately enhanced by inhibition of Delta-Notch signaling as well as by co-expressing the transcription factor Myt1, but not MafA and Pdx1.

Conclusions/interpretation: The results provide further insight into the plasticity of adult human duct cells and suggest measurable routes to enhance Ngn3-mediated in vitro reprogramming protocols for regenerative beta cell therapy in diabetes.

Show MeSH
Related in: MedlinePlus