A method for the generation of human stem cell-derived alpha cells

Peterson, Quinn P.; Veres, Adrian; Chen, Lihua; Slama, Michael; Kenty, Jennifer Hyoje-Ryu; Hassoun, Shaimaa; Brown, Matthew; Dou, Haiqiang; Duffy, Caden D.; Zhou, Quan; Matveyenko, Aleksey V.; Tyrberg, Björn; Sörhede-Winzell, Maria; Rorsman, Patrik; Melton, Douglas A.

doi:10.1038/s41467-020-16049-3

Cited by 65 publications

(60 citation statements)

References 67 publications

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“…To date, only two papers have reported methods of generation of pancreatic alpha cells from hPSCs [ 21 , 22 ]. Rezania et al first reported functional GCG secreting cells produced from human ES cells.…”

Section: Discussionmentioning

confidence: 99%

“…Hyperglycemia in diabetic model mice has been ameliorated by transplanting hPSC-derived pancreatic beta cells into them [ 16 , 17 , 20 ]. Although numerous papers have described methods for hPSCs-beta cell differentiation, only two papers describing differentiation protocols for pancreatic alpha cells from hPSCs are reported; one is from Kieffer’s group [ 21 ] and the other is very recently from Melton`s group [ 22 ]. NKX6.1 expression was shown to be important in determining cell fate between pancreatic beta and alpha cells; cells expressing high levels of NKX6.1 differentiated into pancreatic beta cells, but cells expressing low levels differentiated into pancreatic alpha cells [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

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Efficient induction of pancreatic alpha cells from human induced pluripotent stem cells by controlling the timing for BMP antagonism and activation of retinoic acid signaling

et al. 2021

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Diabetes mellitus is caused by breakdown of blood glucose homeostasis, which is maintained by an exquisite balance between insulin and glucagon produced respectively by pancreatic beta cells and alpha cells. However, little is known about the mechanism of inducing glucagon secretion from human alpha cells. Many methods for generating pancreatic beta cells from human pluripotent stem cells (hPSCs) have been reported, but only two papers have reported generation of pancreatic alpha cells from hPSCs. Because NKX6.1 has been suggested as a very important gene for determining cell fate between pancreatic beta and alpha cells, we searched for the factors affecting expression of NKX6.1 in our beta cell differentiation protocols. We found that BMP antagonism and activation of retinoic acid signaling at stage 2 (from definitive endoderm to primitive gut tube) effectively suppressed NKX6.1 expression at later stages. Using two different hPSCs lines, treatment with BMP signaling inhibitor (LDN193189) and retinoic acid agonist (EC23) at Stage 2 reduced NKX6.1 expression and allowed differentiation of almost all cells into pancreatic alpha cells in vivo after transplantation under a kidney capsule. Our study demonstrated that the cell fate of pancreatic cells can be controlled by adjusting the expression level of NKX6.1 with proper timing of BMP antagonism and activation of retinoic acid signaling during the pancreatic differentiation process. Our method is useful for efficient induction of pancreatic alpha cells from hPSCs.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Efficient induction of pancreatic alpha cells from human induced pluripotent stem cells by controlling the timing for BMP antagonism and activation of retinoic acid signaling

et al. 2021

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“…Current hPSC differentiation protocols permitted the generation of both insulin-secreting β-cells and glucagon-secreting α-cells in a pancreatic endocrine lineage by using molecules to control signaling pathways, as shown in Figure 1 [ 2 , 3 , 7 , 8 , 62 ]. For instance, applying LDN193189 (LDN), an inhibitor of BMP signaling, to the middle stage of the differentiation process could suppress the expression of NKX6.1 and induce the development of α-cells [ 63 ] ( Figure 1 , Table 2 ). As a result, cells were secreting both insulin and glucagon at the end of hESC differentiation [ 7 , 8 ].…”

Section: Molecules Critical For the Generation Of Functional α-Celmentioning

confidence: 99%

“…In addition, phorbol 12,13-dibutyrate (PDBu) is an activator of protein kinase C (PKC). The addition of PDBu at the later stage of hPSC differentiation could significantly increase the percentage of mono-hormonal glucagon-secreting α-cells [ 63 ] ( Figure 1 , Table 2 ). This result proved that the activation of PKC is important for the differentiation of functional α-cells.…”

Section: Molecules Critical For the Generation Of Functional α-Celmentioning

confidence: 99%

Signaling Molecules Regulating Pancreatic Endocrine Development from Pluripotent Stem Cell Differentiation

Huang

Bader

Jin

2020

IJMS

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Diabetes is one of the leading causes of death globally. Currently, the donor pancreas is the only source of human islets, placing extreme constraints on supply. Hence, it is imperative to develop renewable islets for diabetes research and treatment. To date, extensive efforts have been made to derive insulin-secreting cells from human pluripotent stem cells with substantial success. However, the in vitro generation of functional islet organoids remains a challenge due in part to our poor understanding of the signaling molecules indispensable for controlling differentiation pathways towards the self-assembly of functional islets from stem cells. Since this process relies on a variety of signaling molecules to guide the differentiation pathways, as well as the culture microenvironments that mimic in vivo physiological conditions, this review highlights extracellular matrix proteins, growth factors, signaling molecules, and microenvironments facilitating the generation of biologically functional pancreatic endocrine cells from human pluripotent stem cells. Signaling pathways involved in stepwise differentiation that guide the progression of stem cells into the endocrine lineage are also discussed. The development of protocols enabling the generation of islet organoids with hormone release capacities equivalent to native adult islets for clinical applications, disease modeling, and diabetes research are anticipated.

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“…Work from several groups provided cellular atlases of the pancreas during mouse development (Stanescu et al , 2017; Byrnes et al , 2018; Scavuzzo et al , 2018), in adult mice (Muraro et al , 2016) and in human fetal and adult pancreas (Liu et al , 2014; Baron et al , 2016; Muraro et al , 2016; Segerstolpe et al , 2016; Wang et al , 2016; Enge et al , 2017; Han et al , 2020). Efforts have also been made to map cellular identity during pancreas development starting from human pluripotent stem cells (Rezania et al , 2014; Hrvatin et al , 2014; Zhu et al , 2016; Han et al , 2020; Hogrebe et al , 2020; Peterson et al , 2020). Taken together, these studies provide an opportunity to better understand the maintenance and establishment of cellular identity among different pancreatic cell types.…”

Section: Introductionmentioning

confidence: 99%

Predicting the Key Regulators of Cell Identity in Human Adult Pancreas

Vanheer¹,

Schiavo

Haele

et al. 2020

Preprint

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SUMMARYCellular identity during development is under the control of transcription factors that form gene regulatory networks. However, the transcription factors and gene regulatory networks underlying cellular identity in the human adult pancreas remain largely unexplored. Here, we integrate multiple single-cell RNA sequencing datasets of the human adult pancreas, totaling 7393 cells, and comprehensively reconstruct gene regulatory networks. We show that a network of 142 transcription factors forms distinct regulatory modules that characterize pancreatic cell types. We present evidence that our approach identifies key regulators of cell identity in the human adult pancreas. We predict that HEYL and JUND are active in acinar and alpha cells, respectively, and show that these proteins are present in the human adult pancreas as well as in human induced pluripotent stem cell-derived pancreatic cells. The comprehensive gene regulatory network atlas can be explored interactively online. We anticipate our analysis to be the starting point for a more sophisticated dissection of how transcription factors regulate cell identity in the human adult pancreas. Furthermore, given that transcription factors are major regulators of embryo development and are often perturbed in diseases, a comprehensive understanding of how transcription factors work will be relevant in development and disease biology.HIGHLIGHTS-Reconstruction of gene regulatory networks for human adult pancreatic cell types-An interactive resource to explore and visualize gene expression and regulatory states-Predicting putative transcription factors driving pancreatic cell identity-HEYL and JUND as candidate regulators of acinar and alpha cell identity, respectively

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A method for the generation of human stem cell-derived alpha cells

Cited by 65 publications

References 67 publications

Efficient induction of pancreatic alpha cells from human induced pluripotent stem cells by controlling the timing for BMP antagonism and activation of retinoic acid signaling

Efficient induction of pancreatic alpha cells from human induced pluripotent stem cells by controlling the timing for BMP antagonism and activation of retinoic acid signaling

Signaling Molecules Regulating Pancreatic Endocrine Development from Pluripotent Stem Cell Differentiation

Predicting the Key Regulators of Cell Identity in Human Adult Pancreas

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