Islet Neogenesis Associated Protein (INGAP) induces the differentiation of an adult human pancreatic ductal cell line into insulin-expressing cells through stepwise activation of key transcription factors for embryonic beta cell development

Assouline-Thomas, Béatrice; Ellis, Daniel G.; Petropavlovskaia, Maria; Makhlin, Julia; Ding, Jiawang; Rosenberg, Lawrence

doi:10.1016/j.diff.2015.10.008

Cited by 19 publications

(15 citation statements)

References 76 publications

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“…Similarly, human PDCs immortalized by overexpression of E6/E7 genes from human papillomavirus were developed as a surrogate for normal PDCs because of similar genotypic and phenotypic profiles [Yang et al 1998; Ouyang et al 2000]. Incubation of these human pancreatic ductal epithelial (HPDE) cell lines with islet neogenesis associated protein (INGAP) favored the upregulation of β-cell markers [Assouline-Thomas et al 2015]. After short exposure of a week with INGAP, HPDE cells showed the capacity to secrete insulin in response to glucose, albeit the values observed in basal (5.8 mmol of glucose) and stimulated conditions (25 mmol of glucose) were not statistically significant.…”

Section: In Vitro Reprogramming Of Pancreatic Ductal Cells Into β Cellsmentioning

confidence: 99%

β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells

Corritore

Lee

Sokal

et al. 2016

Therapeutic Advances in Endocrinology

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Thorough research on the capacity of human islet transplantation to cure type 1 diabetes led to the achievement of 3-to 5-year-long insulin independence in nearly half of transplanted patients. Yet, translation of this technique to clinical routine is limited by organ shortage and the need for long-term immunosuppression, restricting its use to adults with unstable disease. The production of new bona fide β cells in vitro was thus investigated and finally achieved with human pluripotent stem cells (PSCs). Besides ethical concerns about the use of human embryos, studies are now evaluating the possibility of circumventing the spontaneous tumor formation associated with transplantation of PSCs. These issues fueled the search for cell candidates for β-cell engineering with safe profiles for clinical translation. In vivo studies revealed the regeneration capacity of the exocrine pancreas after injury that depends at least partially on facultative progenitors in the ductal compartment. These stimulated subpopulations of pancreatic ductal cells (PDCs) underwent β-cell transdifferentiation through reactivation of embryonic signaling pathways. In vitro models for expansion and differentiation of purified PDCs toward insulin-producing cells were described using cocktails of growth factors, extracellular-matrix proteins and transcription factor overexpression. In this review, we will describe the latest findings in pancreatic β-cell mass regeneration due to adult ductal progenitor cells. We will further describe recent advances in human PDC transdifferentiation to insulin-producing cells with potential for clinical translational studies.

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Section: In Vitro Reprogramming Of Pancreatic Ductal Cells Into β Cellsmentioning

confidence: 99%

β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells

Corritore

Lee

Sokal

et al. 2016

Therapeutic Advances in Endocrinology

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“…In summary, INGAP has been known for its regenerative properties and the ability to reverse diabetes in animal models 10 , 13 , 15 , 18 , 20 , 71 . Our work shows that INGAP also protects β cells from cytokine-induced apoptosis, thus providing new insights into the anti-diabetic potential of INGAP.…”

Section: Discussionmentioning

confidence: 99%

“…INGAP, a 16.8 kDa Reg3δ protein, was discovered in a partially obstructed hamster pancreas as a factor inducing duct-associated islet neogenesis 8 – 12 . Several studies have indicated that the islet neogenic activity of INGAP protein is mediated by a pentadecapeptide (Ingap-p, aa104–118) 11 , 13 – 16 , which achieves the same effect, albeit at higher concentrations 11 , 16 – 18 . Importantly, Ingap-p has been proven safe to use in humans and has reached Phase 2 clinical trials.…”

Section: Introductionmentioning

confidence: 99%

“…NF-κB activation by IL-1β and IFNγ leads to stimulation of inducible nitric oxide synthase (iNOS) and production of nitric oxide (NO) directly implicated in β-cell apoptosis 46 . To characterize the anti-cytokine effects of INGAP and to investigate the underlying signaling events we used, as previously 17 , 18 , both INGAP-peptide and the full-length recombinant protein (rINGAP).…”

Section: Introductionmentioning

confidence: 99%

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Islet neogenesis associated protein (INGAP) protects pancreatic β cells from IL-1β and IFNγ-induced apoptosis

2021

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The goal of this study was to determine whether recombinant Islet NeoGenesis Associated Protein (rINGAP) and its active core, a pentadecapeptide INGAP104–118 (Ingap-p), protect β cells against cytokine-induced death. INGAP has been shown to induce islet neogenesis in diabetic animals, to stimulate β-cell proliferation and differentiation, and to improve islet survival and function. Importantly, Ingap-p has shown promising results in clinical trials for diabetes (phase I/II). However, the full potential of INGAP and its mechanisms of action remain poorly understood. Using rat insulinoma cells RINm5F and INS-1 treated with interleukin-1β (IL-1β) and interferon‐gamma (IFN‐γ), we demonstrate here that both rINGAP and Ingap-p inhibit apoptosis, Caspase-3 activation, inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production, and explore the related signaling pathways. As expected, IL-1β induced nuclear factor kappa B (NF-κB), p38, and JNK signaling, whereas interferon‐gamma (IFN‐γ) activated the JAK2/STAT1 pathway and potentiated the IL-1β effects. Both rINGAP and Ingap-p decreased phosphorylation of IKKα/β, IkBα, and p65, although p65 nuclear translocation was not inhibited. rINGAP, used for further analysis, also inhibited STAT3, p38, and JNK activation. Interestingly, all inhibitory effects of rINGAP were observed for the cytokine cocktail, not IL-1β alone, and were roughly equal to reversing the potentiating effects of INFγ. Furthermore, rINGAP had no effect on IL-1β/NF-κB-induced gene expression (e.g., Ccl2, Sod2) but downregulated several IFNγ-stimulated (Irf1, Socs1, Socs3) or IFNγ-potentiated (Nos2) genes. This, however, was observed again only for the cytokine cocktail, not IFNγ alone, and rINGAP did not inhibit the IFNγ-induced JAK2/STAT1 activation. Together, these intriguing results suggest that INGAP does not target either IL-1β or IFNγ individually but rather inhibits the signaling crosstalk between the two, the exact mechanism of which remains to be investigated. In summary, our study characterizes the anti-inflammatory effects of INGAP, both protein and peptide, and suggests a new therapeutic utility for INGAP in the treatment of diabetes.

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“…The mechanisms underlay islet-neogenesis are complexes and involve signaling and cell-cycle regulators, pancreatic islet specific transcription factors, and growth factors [ 50 , 51 , 52 ]. In particular, human fibroblast growth factor (FGF)-2b signaling has been implicated in patterning proliferation and cell differentiation in many organs including the developing pancreas [ 53 , 54 ].…”

Section: Introductionmentioning

confidence: 99%

FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes

Donadel

Pastore

Della-Morte

et al. 2017

IJMS

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Background: Diabetes mellitus (DM) is a multifactorial disease orphan of a cure. Regenerative medicine has been proposed as novel strategy for DM therapy. Human fibroblast growth factor (FGF)-2b controls β-cell clusters via autocrine action, and human placental lactogen (hPL)-A increases functional β-cells. We hypothesized whether FGF-2b/hPL-A treatment induces β-cell differentiation from ductal/non-endocrine precursor(s) by modulating specific genes expression. Methods: Human pancreatic ductal-cells (PANC-1) and non-endocrine pancreatic cells were treated with FGF-2b plus hPL-A at 500 ng/mL. Cytofluorimetry and Immunofluorescence have been performed to detect expression of endocrine, ductal and acinar markers. Bromodeoxyuridine incorporation and annexin-V quantified cells proliferation and apoptosis. Insulin secretion was assessed by RIA kit, and electron microscopy analyzed islet-like clusters. Results: Increase in PANC-1 duct cells de-differentiation into islet-like aggregates was observed after FGF-2b/hPL-A treatment showing ultrastructure typical of islets-aggregates. These clusters, after stimulation with FGF-2b/hPL-A, had significant (p < 0.05) increase in insulin, C-peptide, pancreatic and duodenal homeobox 1 (PDX-1), Nkx2.2, Nkx6.1, somatostatin, glucagon, and glucose transporter 2 (Glut-2), compared with control cells. Markers of PANC-1 (Cytokeratin-19, MUC-1, CA19-9) were decreased (p < 0.05). These aggregates after treatment with FGF-2b/hPL-A significantly reduced levels of apoptosis. Conclusions: FGF-2b and hPL-A are promising candidates for regenerative therapy in DM by inducing de-differentiation of stem cells modulating pivotal endocrine genes.

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Islet Neogenesis Associated Protein (INGAP) induces the differentiation of an adult human pancreatic ductal cell line into insulin-expressing cells through stepwise activation of key transcription factors for embryonic beta cell development

Cited by 19 publications

References 76 publications

β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells

β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells

Islet neogenesis associated protein (INGAP) protects pancreatic β cells from IL-1β and IFNγ-induced apoptosis

FGF-2b and h-PL Transform Duct and Non-Endocrine Human Pancreatic Cells into Endocrine Insulin Secreting Cells by Modulating Differentiating Genes

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