β-Cell Differentiation from a Human Pancreatic Cell Line in Vitro and in Vivo

Tour, D. Dufayet De La; Halvorsen, Tanya; Demeterco, Carla; Tyrberg, Björn; Itkin-Ansari, Pamela; Loy, Mary; Yoo, Soon‐Jib; Hao, Ergeng; Bossie, Stuart; Levine, Fred

doi:10.1210/mend.15.3.0604

Cited by 55 publications

(20 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…By inducing expression of PDX-1 in the cells using a human PDX-1 expression vector we also induced transcription of the insulin gene. This confirms that PDX-1 serves as a master control switch for induction of b-cell phenotype and that the induction of PDX-1 (Stoffers et al, 1997;Dufayet de la Tour et al, 2001), in turn, is the critical step in EX-4-mediated differentiation to an endocrine phenotype. As a consequence, insulin was synthesized and subsequently secreted in a regulated manner.…”

Section: Discussionsupporting

confidence: 66%

Exendin‐4 differentiation of a human pancreatic duct cell line into endocrine cells: Involvement of PDX‐1 and HNF3β transcription factors

Zhou

Piñeyro

Wang

et al. 2002

Journal Cellular Physiology

View full text Add to dashboard Cite

Exendin-4 (EX-4), a long acting agonist of GLP-1, induces an endocrine phenotype in Capan-1 cells. Under culture conditions which include serum, approximately 10% of the cells contain insulin and glucagon. When exposed to EX-4 (0.1 nM, up to 5 days), the number of cells containing insulin and glucagon increased to approximately 40%. Western blot analysis detected a progressive increase in protein levels of glucokinase and GLUT2 over 3 days of EX-4 treatment. We explored the sequence of activation of certain transcription factors known to be essential for the beta cell phenotype: PDX-1, Beta2/NeuroD, and hepatocyte nuclear factor 3beta (HNF3beta). Double immunostaining showed that PDX-1 coexisted with insulin and glucagon in EX-4-treated cells. Treatment caused an increase in PDX-1 protein levels by 24 h and induced its nuclear translocation. Beta2/NeuroD protein levels also increased progressively over 24 h. HNF3beta protein level increased twofold as early as 6 h after EX-4 treatment. EMSA results indicated that EX-4 caused a 12-fold increase in HNF3beta binding to PDX-1 promoter area II. Beta2/NeuroD protein levels progressively increased after 24 h treatment. Differentiation to insulin-producing cells was also seen when Capan-1 cells were transfected with pdx-1, with 80% of these cells expressing insulin 3 days after transfection. PDX-1 antisense totally inhibited such conversion. During the differentiation of duct cells to endocrine cells, cAMP levels (EX-4 is a ligand for the GLP-1, G-protein coupled receptor) and MAP kinase activity increased. Our results indicate that EX-4 activates adenylyl cyclase and MAP kinase which, in turn, may lead to activation of transcription factors necessary for an endocrine phenotype.

show abstract

Section: Discussionsupporting

confidence: 66%

Exendin‐4 differentiation of a human pancreatic duct cell line into endocrine cells: Involvement of PDX‐1 and HNF3β transcription factors

Zhou

Piñeyro

Wang

et al. 2002

Journal Cellular Physiology

View full text Add to dashboard Cite

show abstract

“…The human Persistant Hyperinsulinemic Hypoglycemia of Infancy (PHHI)-derived pancreatic beta cell line, NES2Y, has also been reported to exhibit glucosestimulated insulin secretion after successful transfection to repair defects in expression of K-ATP channel and PDX-1 genes (17). Blox5 is an immortalized cell line produced from a purified population of human beta cells by infection with retroviral vectors expressing the SV40 T antigen, H-ras val12 , and hTERT oncogenes (18). This cell line grows indefinitely but loses differentiated function and the ability to express insulin.…”

mentioning

confidence: 99%

Development and Functional Characterization of Insulin-releasing Human Pancreatic Beta Cell Lines Produced by Electrofusion

McCluskey

Hamid

Guo-Parke

et al. 2011

Journal of Biological Chemistry

132

103

View full text Add to dashboard Cite

Three novel human insulin-releasing cell lines designated 1.1B4, 1.4E7, and 1.1E7 were generated by electrofusion of freshly isolated of human pancreatic beta cells and the immortal human PANC-1 epithelial cell line. Functional studies demonstrated glucose sensitivity and responsiveness to known modulators of insulin secretion. Western blot, RT-PCR, and immunohistochemistry showed expression of the major genes involved in proinsulin processing and the pancreatic beta cell stimulussecretion pathway including PC1/3, PC2, GLUT-1, glucokinase, and K-ATP channel complex (Sur1 and Kir6.2) and the voltagedependent L-type Ca 2؉ channel. The cells stained positively for insulin, and 1.1B4 cells were used to demonstrate specific staining for insulin, C-peptide, and proinsulin together with insulin secretory granules by electron microscopy. Analysis of metabolic function indicated intact mechanisms for glucose uptake, oxidation/utilization, and phosphorylation by glucokinase. Glucose, alanine, and depolarizing concentrations of K ؉ were all able to increase [Ca 2؉ ] i in at least two of the cell lines tested. Insulin secretion was also modulated by other nutrients, hormones, and drugs acting as stimulators or inhibitors in normal beta cells. Subscapular implantation of the 1.1B4 cell line improved hyperglycemia and resulted in glucose lowering in streptozotocin-diabetic SCID mice. These novel human electrofusion-derived beta cell lines therefore exhibit stable characteristics reminiscent of normal pancreatic beta cells, thereby providing an unlimited source of human insulin-producing cells for basic biochemical studies and pharmacological drug testing plus proof of concept for cellular insulin replacement therapy.

show abstract

“…Furthermore, GLP-1 and exendin-4 promote differentiation of exocrine cell lines toward a ␤ cell phenotype (21), a process that appears to depend on the expression of Pdx-1 (22,23).…”

mentioning

confidence: 99%

Glucagon-like Peptide-1 Receptor Signaling Modulates β Cell Apoptosis

Li¹,

Hansotia²,

Yusta³

et al. 2003

Journal of Biological Chemistry

555

379

View full text Add to dashboard Cite

Glucagon-like peptide-1 (GLP-1) stimulates insulin secretion and augments ␤ cell mass via activation of ␤ cell proliferation and islet neogenesis. We examined whether GLP-1 receptor signaling modifies the cellular susceptibility to apoptosis. Mice administered streptozotocin (STZ), an agent known to induce ␤ cell apoptosis, exhibit sustained improvement in glycemic control and increased levels of plasma insulin with concomitant administration of the GLP-1 agonist exendin-4 (Ex-4). Blood glucose remained significantly lower for weeks after cessation of exendin-4. STZ induced ␤ cell apoptosis, which was significantly reduced by co-administration of Ex-4. Conversely, mice with a targeted disruption of the GLP-1 receptor gene exhibited increased ␤ cell apoptosis after STZ administration. Exendin-4 directly reduced cytokine-induced apoptosis in purified rat ␤ cells exposed to interleukin 1␤, tumor necrosis fator ␣, and interferon ␥ in vitro. Furthermore, Ex-4-treated BHK-GLP-1R cells exhibited significantly increased cell viability, reduced caspase activity, and decreased cleavage of ␤-catenin after treatment with cycloheximide in vitro. These findings demonstrate that GLP-1 receptor signaling directly modifies the susceptibility to apoptotic injury, and provides a new potential mechanism linking GLP-1 receptor activation to preservation or enhancement of ␤ cell mass in vivo.Glucagon-like peptide-1 (GLP-1) 1 is derived from posttranslational processing of proglucagon in enteroendocrine L cells (1) and is secreted from the distal gut after nutrient ingestion (2). The termination of GLP-1 action by the enzyme dipeptidyl peptidase IV occurs within minutes following GLP-1 secretion (3-5), yet GLP-1 exerts several rapid metabolic actions including stimulation and inhibition of insulin and glucagon secretion, respectively (6 -10). GLP-1 action is essential for glucose homeostasis, because GLP-1 receptor blockade with the antagonist exendin (9 -39) increases blood glucose and decreases levels of circulating insulin in human and rodent studies (11)(12)(13)(14).Activation of GLP-1 receptor signaling leads to enhanced expression of mRNA transcripts for glucokinase, GLUT-2, Pdx-1, and insulin in ␤ cell lines (15-17) and in both normal and diabetic rodents (18 -20). Furthermore, GLP-1 and exendin-4 promote differentiation of exocrine cell lines toward a ␤ cell phenotype (21), a process that appears to depend on the expression of Pdx-1 (22, 23).GLP-1 receptor signaling is also coupled to formation of new ␤ cells through enhanced proliferation of existing ␤ cells (24) and via induction of islet neogenesis (25). The mitogenic actions of GLP-1 are detectable in normal rodents (20,24) and in the setting of experimental diabetes (19,25). Administration of GLP-1 or exendin-4 to newborn rats treated with the ␤ cell toxin streptozotocin (STZ) leads to increased ␤ cell mass at postnatal day 7, which persists and remains increased at 2 months of age. The increased ␤ cell mass in the GLP-1/exendin-4 treated rats was attributed to both enhan...

show abstract

β-Cell Differentiation from a Human Pancreatic Cell Line in Vitro and in Vivo

Cited by 55 publications

References 29 publications

Exendin‐4 differentiation of a human pancreatic duct cell line into endocrine cells: Involvement of PDX‐1 and HNF3β transcription factors

Exendin‐4 differentiation of a human pancreatic duct cell line into endocrine cells: Involvement of PDX‐1 and HNF3β transcription factors

Development and Functional Characterization of Insulin-releasing Human Pancreatic Beta Cell Lines Produced by Electrofusion

Glucagon-like Peptide-1 Receptor Signaling Modulates β Cell Apoptosis

Contact Info

Product

Resources

About