Inhibitors of Src and Focal Adhesion Kinase Promote Endocrine Specification

Afrikanova, Ivka; Yebra, Mayra; Simpkinson, Megan; Xu, Yiming; Hayek, Alberto; Montgomery, Anthony M.P.

doi:10.1074/jbc.m111.290825

Cited by 23 publications

(15 citation statements)

References 50 publications

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“…These results are similar to those demonstrating that the selective tyrosine kinase inhibitor imatinib promotes INS release from β cells32. In addition, the blockade of tyrosine kinase signalling, such as Src family kinases and FAK, can promote β cell differentiation33. Thus, there is growing evidence that tyrosine kinase signalling has a role in the differentiation of β cells as well as in maintaining the function of mature β cells.…”

Section: Discussionsupporting

confidence: 85%

An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors

Yamashita‐Sugahara

Matsumoto

Ohtaka

et al. 2016

Sci Rep

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Human induced pluripotent stem cells (hiPSCs) provide a potential resource for regenerative medicine. To identify the signalling pathway(s) contributing to the development of functional β cells, we established a tracing model consisting of dual knock-in hiPSCs (INS-Venus/NGN3-mCherry) (hIveNry) expressing the fluorescent proteins Venus and mCherry under the control of intrinsic insulin (INS) and neurogenin 3 (NGN3) promoters, respectively. hIveNry iPSCs differentiated into NGN3- and mCherry-positive endocrine progenitors and then into Venus-positive β cells expressing INS, PDX1, NKX6.1, and glucokinase (GCK). Using these cells, we conducted high-throughput screening of chemicals and identified a specific kinase inhibitor of fibroblast growth factor receptor 1 (FGFR1) that acted in a stage-dependent manner to promote the terminal differentiation of pancreatic endocrine cells, including β cells, from the intermediate stage of pancreatic endocrine progenitors while blocking the early development of pancreatic progenitors. This FGFR1 inhibitor augmented the expression of functional β cell markers (SLC30A8 and ABCC8) and improved glucose-stimulated INS secretion. Our findings indicate that the hIveNry model could provide further insights into the mechanisms of hiPS-derived β cell differentiation controlled by FGFR1-mediated regulatory pathways in a temporal-dependent fashion.

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Section: Discussionsupporting

confidence: 85%

An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors

Yamashita‐Sugahara

Matsumoto

Ohtaka

et al. 2016

Sci Rep

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“…Half of the spheres were transferred to one plate and kept in a growth medium (control), and the other half were kept in a differentiation medium, which contained DMEM high glucose, B27 supplement and PP2, a Src family kinase inhibitor, since we recently showed that Src family kinase inhibition is involved in endocrine specification. 27 After 4 days, the spheres were harvested and analyzed for pancreatic markers of progenitor and hormone-expressing cells. We found robust expression of Pdx1 , Nkx6.1 , insulin, glucagon, and somatostatin in SSEA4 + in comparison to the controls and to SSEA4 − cell-derived spheres (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…mRNA purification, cDNA synthesis, and real-time quantitative polymerase chain reaction (RT-qPCR) analysis of differentiated tissue were performed as previously described. 27 Briefly, 20 μL PCR reactions were run using 3 μL of cDNA, combined with the TaqMan Universal PCR Master Mix (4324018; Applied Biosystems) and unlabeled PCR primers and a TaqMan FAM ™ dye-labeled probe listed in Supplementary Table S1. RT-qPCR was performed using an Applied Biosystems StepOnePlus real time PCR machine.…”

Section: Methodsmentioning

confidence: 99%

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Is Stage-Specific Embryonic Antigen 4 a Marker for Human Ductal Stem/Progenitor Cells?

Afrikanova

Kayali

Lopez

et al. 2012

BioResearch Open Access

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The presence of pancreatic stem cells (PnSCs) has not been firmly demonstrated in the human or animal pancreas. Previous reports have suggested that ductal and acinar structures in the exocrine pancreas can be a potential source of progenitor cells. More recently, immature insulin precursors in the periphery of human islets have been found to self-replicate and differentiate to endocrine cells in vitro. Transplantation of these cells under the kidney capsule improves the diabetic state in mice. The controversy surrounding where PnSCs reside could be resolved if a specific marker were to be found that allowed their identification, purification, and directed differentiation to endocrine cells. We have identified in human pancreas cells positive for the stage-specific embryonic antigen 4 (SSEA4), a stem cell marker. These cells also express ductal, pancreatic progenitor, and stem cell protein markers. Interestingly, some of the SSEA4+ cells scattered in the ducts do not show a ductal cell phenotype. SSEA4+-sorted cells formed aggregate-like spheres in culture and robustly differentiated to pancreatic hormone-expressing cells in conditions of high glucose concentration and B27 supplementation. We hypothesize that SSEA4+ cells or a subpopulation of those cells residing in the pancreatic ducts may be the elusive PnSCs, and in this case, SSEA4 may represent a potential surface antigen marker for human PnSCs. The discovery of specific markers for the identification and purification of human PnSCs would greatly facilitate studies aimed at the expansion of these cells and the development of targeting tools for their potential induction to insulin-producing cells.

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“…The self-organization of endocrine progenitors into islet structure appears to be mediated by G-protein coupled receptors, non-canonical Wnt, and receptor tyrosine kinase (RTK) signaling [45]. While the importance of delamination and migration to in vitro derivation of endocrine cells from hPSC is not known, endocrine differentiation is promoted in the presence of focal adhesion kinase inhibitor, suggesting that modulation of EMT-related pathways may control endocrine specification in hPSC culture [46]. …”

Section: Cell Migration Creates Isletsmentioning

confidence: 99%

Tissue engineering 2.0: guiding self-organization during pluripotent stem cell differentiation

Woodford

Zandstra

2012

Current Opinion in Biotechnology

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Human pluripotent stem cell (hPSC) differentiation aims to mimic development using growth factors or small molecules in a time- and dose-dependent manner. However, the cell types produced using this approach are predominantly fetal-like in phenotype and function, limiting their use in regenerative medicine. This is particularly true in current efforts to produce pancreatic beta cells, wherein robust pancreatic progenitor maturation can only be accomplished upon transplantation into mice. Recent studies have suggested that hPSC-derived cells are capable of self-organizing in vitro, revealing a new paradigm for creating mature cells and tissues. Tissue engineering strategies that provide subtle and dynamic signals to developmentally naïve cells may be applied to mimic in vitro the self-organization aspects of pancreatic development.

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Inhibitors of Src and Focal Adhesion Kinase Promote Endocrine Specification

Cited by 23 publications

References 50 publications

An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors

An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors

Is Stage-Specific Embryonic Antigen 4 a Marker for Human Ductal Stem/Progenitor Cells?

Tissue engineering 2.0: guiding self-organization during pluripotent stem cell differentiation

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