In vivo reprogramming of pancreatic acinar cells to three islet endocrine subtypes

Li, Weida; Nakanishi, Mio; Zumsteg, Adrian; Shear, Matthew A.; Wright, Christopher V.E.; Melton, Douglas A.; Zhou, Qiao

doi:10.7554/elife.01846

Cited by 123 publications

(105 citation statements)

References 51 publications

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“…This implies that a small number of genes control cell identify. This is perhaps not surprising because it has previously been shown that only three transcription factors control endocrine cell fate (19,20). Our transcription factor analysis revealed few differences in their expression between α-and β-cells, as well as between α-cells and PP cells.…”

Section: Discussionsupporting

confidence: 76%

Use of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells

Xin

Kim

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

152

129

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This study provides an assessment of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells. The system combines microfluidic technology and nanoliter-scale reactions. We sequenced 622 cells, allowing identification of 341 islet cells with high-quality gene expression profiles. The cells clustered into populations of α-cells (5%), β-cells (92%), δ-cells (1%), and pancreatic polypeptide cells (2%). We identified cell-type-specific transcription factors and pathways primarily involved in nutrient sensing and oxidation and cell signaling. Unexpectedly, 281 cells had to be removed from the analysis due to low viability, low sequencing quality, or contamination resulting in the detection of more than one islet hormone. Collectively, we provide a resource for identification of high-quality gene expression datasets to help expand insights into genes and pathways characterizing islet cell types. We reveal limitations in the C1 Fluidigm cell capture process resulting in contaminated cells with altered gene expression patterns. This calls for caution when interpreting single-cell transcriptomics data using the C1 Fluidigm system.

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Section: Discussionsupporting

confidence: 76%

Use of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells

Xin

Kim

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

152

129

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“…The adult pancreatic acinar cell, previously thought to be terminally and irreversibly differentiated, is now known to be surprisingly malleable (87)(88)(89)(90). We showed that two critical aspects of regulation by Ptf1a define the acinar cell identity.…”

Section: Discussionmentioning

confidence: 88%

Transcriptional Maintenance of Pancreatic Acinar Identity, Differentiation, and Homeostasis by PTF1A

Hoang

Hale

Azevedo-Pouly

et al. 2016

Molecular and Cellular Biology

113

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Maintenance of cell type identity is crucial for health, yet little is known of the regulation that sustains the long-term stability of differentiated phenotypes. To investigate the roles that key transcriptional regulators play in adult differentiated cells, we examined the effects of depletion of the developmental master regulator PTF1A on the specialized phenotype of the adult pancreatic acinar cell in vivo. Transcriptome sequencing and chromatin immunoprecipitation sequencing results showed that PTF1A maintains the expression of genes for all cellular processes dedicated to the production of the secretory digestive enzymes, a highly attuned surveillance of unfolded proteins, and a heightened unfolded protein response (UPR). Control by PTF1A is direct on target genes and indirect through a ten-member transcription factor network. Depletion of PTF1A causes an imbalance that overwhelms the UPR, induces cellular injury, and provokes acinar metaplasia. Compromised cellular identity occurs by derepression of characteristic stomach genes, some of which are also associated with pancreatic ductal cells. The loss of acinar cell homeostasis, differentiation, and identity is directly relevant to the pathologies of pancreatitis and pancreatic adenocarcinoma. Loss of cellular identity has long been associated with tissue injury and a first step in cancer progression (for examples, see references 1 and 2). Maintenance of a specific cellular phenotype depends on the continued transcription of cell-type-specific genes, largely through open chromatin architecture (3, 4) maintained by a small group of lineage-restricted DNA-binding transcription factors (TFs) (5, 6) that establish a unique transcriptional regulatory network (7). Many physiologic or pathophysiologic perturbations can affect the differentiated state of a cell quantitatively, but fewer affect the state of differentiation qualitatively. Qualitative changes involve the acquisition of characteristics of another cell type (or types), often defined by one or a few cell-specific markers, in addition to the diminution of the original phenotype. Despite progress with cellular reprogramming (for example, see reference 8), the molecular and genetic mechanisms that maintain cellular identity within the context of adult organs remain incompletely understood. In this report, we show that inactivation of the transcriptional regulatory gene Ptf1a in adult pancreatic acinar cells has pleiotropic effects on gene expression that cause quantitative and multigene qualitative changes of acinar differentiation.The acinar cell of the pancreas has been an informative model of terminal cellular differentiation (9). Common cellular processes are greatly exaggerated in support of the prodigious synthesis, processing, storage, and exocytosis of secretory proteins. The pancreatic acinar cell has the most ribosomes (10) and the highest rate of protein synthesis (11) of any mammalian somatic cell; it synthesizes, stores, and secretes its weight in protein daily. Specialized cellular functions ...

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“…Endogenous mouse pancreatic acinar cells transdifferentiate into insulin-producing b cells on overexpression of NGN3, MAFA, and PDX1. 18,19 Subsets of this transcription factor cocktail, NGN3 þ MAFA, and NGN3 alone, successfully induce related pancreatic cell subtypes such as glucagon þ a cells and somatostatin þ d cells. 19 Outside of the pancreas, expression of the proeb-cell transcription factors NGN3, MAFA, and PDX3 generates insulin-producing b-like cells from liver hepatocytes.…”

Section: Fate-specific In Vitro Reprogramming Methodsmentioning

confidence: 99%

“…19 The misexpression of Ngn3, Ngn3 þ Mafa or Ngn3 þ Mafa þ Pdx1 reprograms pancreatic acinar cells to somatostatin þ d cells, glucagon þ a cells or insulin þ b cells, respectively. Ultrastructural analysis of secretory granules, hormone secretion, cell-specific marker expression, and modifications to patterned DNA methylation indicate both phenotypic and functional maturity after cell identity conversion.…”

Section: Reprogramming Pancreatic Acinar Cells and Hepatocytesmentioning

confidence: 99%

Regeneration through Reprogramming Adult Cell Identity in Vivo

Zhang

2015

The American Journal of Pathology

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The discovery and in vivo application of cell fate reprogramming concepts have jumpstarted new technologies aimed at the functional regeneration of damaged tissues. As most adult organ systems retain only a limited potential for self-regeneration after trauma, the production of fate-specific cells by in vivo transdifferentiation offers a targeted method for tissue bioengineering. Proof-of-principle studies have demonstrated the induction of neural precursor cells, neurons, cardiomyocytes, and insulin-producing β islet cells. Each of these induced cell types survive, mature, and integrate into the local environment in a functionally meaningful manner. Here, we briefly highlight recent advances in the in vivo reprogramming of cell identity and the current challenges that face the clinical relevance of these methods.

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In vivo reprogramming of pancreatic acinar cells to three islet endocrine subtypes

Cited by 123 publications

References 51 publications

Use of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells

Use of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells

Transcriptional Maintenance of Pancreatic Acinar Identity, Differentiation, and Homeostasis by PTF1A

Regeneration through Reprogramming Adult Cell Identity in Vivo

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