Overexpression of PAX4 reduces glucagon expression in differentiating hESCs

Gage, Blair K.; Baker, Robert K.; Kieffer, Timothy J.

doi:10.4161/isl.29236

Cited by 24 publications

(22 citation statements)

References 36 publications

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“…Adenoviral infection of differentiating hESCs (WT and ARX ko) occurred on days 13 and 19 of differentiation (24 hours, MOI of 2 based on 1 x 10 6 cells / well). eGFP was visualized using an Axiovert 200 microscope (Carl Zeiss Canada, Toronto, ON, Canada) as previously described [ 28 ]. Transgene delivery was assessed 72 hours after infection based on eGFP fluorescence by flow cytometry using a LSRII cytometer (Becton Dickinson, San Jose, CA, USA) and FlowJo Software (Tree Star, Ashland, OR, USA) (n = 3).…”

Section: Methodsmentioning

confidence: 99%

The Role of ARX in Human Pancreatic Endocrine Specification

et al. 2015

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The in vitro differentiation of human embryonic stem cells (hESCs) offers a model system to explore human development. Humans with mutations in the transcription factor Aristaless Related Homeobox (ARX) often suffer from the syndrome X-linked lissencephaly with ambiguous genitalia (XLAG), affecting many cell types including those of the pancreas. Indeed, XLAG pancreatic islets lack glucagon and pancreatic polypeptide-positive cells but retain somatostatin, insulin, and ghrelin-positive cells. To further examine the role of ARX in human pancreatic endocrine development, we utilized genomic editing in hESCs to generate deletions in ARX. ARX knockout hESCs retained pancreatic differentiation capacity and ARX knockout endocrine cells were biased toward somatostatin-positive cells (94% of endocrine cells) with reduced pancreatic polypeptide (rarely detected), glucagon (90% reduced) and insulin-positive (65% reduced) lineages. ARX knockout somatostatin-positive cells shared expression patterns with human fetal and adult δ-cells. Differentiated ARX knockout cells upregulated PAX4, NKX2.2, ISL1, HHEX, PCSK1, PCSK2 expression while downregulating PAX6 and IRX2. Re-expression of ARX in ARX knockout pancreatic progenitors reduced HHEX and increased PAX6 and insulin expression following differentiation. Taken together these data suggest that ARX plays a key role in pancreatic endocrine fate specification of pancreatic polypeptide, somatostatin, glucagon and insulin positive cells from hESCs.

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Section: Methodsmentioning

confidence: 99%

The Role of ARX in Human Pancreatic Endocrine Specification

et al. 2015

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“…Further evidence on PAX4 triggering β-cell commitment arises from transgenic mouse models where induction of PAX4 expression in early pancreatic epithelium, or in endocrine committed precursor cells induces the formation of insulin producing β-cells at the expense of all other islet cell phenotypes [25]. In agreement with this, ectopic expression of PAX4 potentiates the in vitro generation of insulin + β-like cells [26,27,28,29,30,31,32]. …”

Section: Pax4 In Islet Physiology: Key Player In β-Cell Generationmentioning

confidence: 98%

The Diabetes-Linked Transcription Factor PAX4: From Gene to Functional Consequences

Lorenzo

Juárez-Vicente

Cobo-Vuilleumier

et al. 2017

Genes

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Paired box 4 (PAX4) is a key factor in the generation of insulin producing β-cells during embryonic development. In adult islets, PAX4 expression is sequestered to a subset of β-cells that are prone to proliferation and more resistant to stress-induced apoptosis. The importance of this transcription factor for adequate pancreatic islets functionality has been manifested by the association of mutations in PAX4 with the development of diabetes, independently of its etiology. Overexpression of this factor in adult islets stimulates β-cell proliferation and increases their resistance to apoptosis. Additionally, in an experimental model of autoimmune diabetes, a novel immunomodulatory function for this factor has been suggested. Altogether these data pinpoint at PAX4 as an important target for novel regenerative therapies for diabetes treatment, aiming at the preservation of the remaining β-cells in parallel to the stimulation of their proliferation to replenish the β-cell mass lost during the progression of the disease. However, the adequate development of such therapies requires the knowledge of the molecular mechanisms controlling the expression of PAX4 as well as the downstream effectors that could account for PAX4 action.

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“…Overexpression of several pancreatic development TFs enhances pancreatic β‐cell differentiation (Soria et al ., ; Blyszczuk et al ., ; Kwon et al ., ). For example, overexpression of the PAX4 TF in hESCs significantly promotes pancreatic β‐cell generation (Liew et al ., ) and increases the number of β cells by suppressing the expression of ARX and glucagon (Gage, Baker & Kieffer, ). Previous studies employed forskolin (an adenylate cyclase activator) and dexamethasone (a synthetic adrenocortical steroid) (Kunisada et al ., ), hepatocyte growth factor (HGF), insulin growth factor 1 (IGF‐1) and GLP‐1 activators (Ohmine et al ., ) to enhance pancreatic β‐cell maturation.…”

Section: Differentiation Of Pluripotent Stem Cells Into Pancreatic β‐mentioning

confidence: 99%

Pathways governing development of stem cell‐derived pancreatic β cells: lessons from embryogenesis

et al. 2017

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The loss of functional β cells leads to development of diabetes. Several studies have shown that β cells are specified through several stages of progenitors during pancreas development, each stage defined by the expression of specific transcription factors (TFs). Understanding signalling pathways that control the differentiation and specification processes during embryogenesis will facilitate efforts to obtain functional β cells in vitro. Our current knowledge of the mechanisms involved in pancreatic β cell development and survival under normal or diabetic conditions has come largely from animal studies. However, there are marked differences in islet structure and physiological properties between humans and animals, and not all phenotypes of human diabetes can be recapitulated in animal models. Therefore, human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced PSCs (hiPSCs) offer a great opportunity for increasing our understanding of the pathways regulating human pancreatic β-cell development and survival. Furthermore, hPSCs provide a renewable source of functional pancreatic β cells for cell replacement therapy as well as disease modelling. Herein, we discuss the signalling pathways involved in the development of pancreatic β cells during embryogenesis. Additionally, we describe how these pathways are manipulated in vitro to differentiate hPSCs into functional β cells. Finally, we highlight the progress that has been made for the applications of those cells in treating and modelling diabetes.

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Overexpression of PAX4 reduces glucagon expression in differentiating hESCs

Cited by 24 publications

References 36 publications

The Role of ARX in Human Pancreatic Endocrine Specification

The Role of ARX in Human Pancreatic Endocrine Specification

The Diabetes-Linked Transcription Factor PAX4: From Gene to Functional Consequences

Pathways governing development of stem cell‐derived pancreatic β cells: lessons from embryogenesis

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