Combination Therapy with Epidermal Growth Factor and Gastrin Induces Neogenesis of Human Islet β-Cells from Pancreatic Duct Cells and an Increase in Functional β-Cell Mass

Abstract

Pancreatic islet transplantation is a viable treatment for type 1 diabetes, but is limited by human donor tissue availability. The combination of epidermal growth factor (EGF) and gastrin induces islet beta-cell neogenesis from pancreatic exocrine duct cells in rodents. In this study we investigated whether EGF and gastrin could expand the beta-cell mass in adult human isolated islets that contain duct as well as endocrine cells. Human islet cells were cultured for 4 wk in serum-free medium (control) or in medium with EGF (0.3 mug/ml), gastrin (1.0 mug/ml), or the combination of EGF and gastrin. beta-Cell numbers were increased in cultures with EGF plus gastrin (+118%) and with EGF (+81%), but not in cultures with gastrin (-3%) or control medium (-62%). After withdrawal of EGF and gastrin and an additional 4 wk in control medium, beta-cell numbers continued to increase only in cultures previously incubated with both EGF and gastrin (+232%). EGF plus gastrin also significantly increased cytokeratin 19-positive duct cells (+678%) in the cultures. Gastrin, alone or in combination with EGF, but not EGF alone, increased the expression of pancreatic and duodenal homeobox factor-1 as well as insulin and C peptide in the cytokeratin 19-positive duct cells. Also, EGF plus gastrin significantly increased beta-cells and insulin content in human islets implanted in immunodeficient nonobese diabetic-severe combined immune deficiency mice as well as insulin secretory responses of the human islet grafts to glucose challenge. In conclusion, combination therapy with EGF and gastrin increases beta-cell mass in adult human pancreatic islets in vitro and in vivo, and this appears to result from the induction of beta-cell neogenesis from pancreatic exocrine duct cells.

In vitro neogenesis of human islets reflects the plasticity of differentiated human pancreatic cells

“…There are fewer observations in human pancreatic cells but clear evidence exists, particularly for the islet/ductal transitional cells both in vivo [6] and in vitro [14,15,31]. However, only a recent report [31] and our study have observed a relatively large number of transitional cells within all four types of endocrine cells and presented the quantitative results while only occasional co-staining of insulin and ductal markers is found by others. Our demonstration of endocrine-ductal transitional cells and the fact that depletion of endocrine cells from the starting material severely reduces the endocrine differentiation of the expanded CK19-positive cells promotes the hypothesis that this particular form of plasticity could be important for human islet expansion.…”

“…Transdifferentiation of acinar cells to ductal cells and further differentiation of these cells into islets has recently been demonstrated in vivo [29] and in vitro [30]. There are fewer observations in human pancreatic cells but clear evidence exists, particularly for the islet/ductal transitional cells both in vivo [6] and in vitro [14,15,31]. However, only a recent report [31] and our study have observed a relatively large number of transitional cells within all four types of endocrine cells and presented the quantitative results while only occasional co-staining of insulin and ductal markers is found by others.…”

Growth factors and medium hyperglycemia induce Sox9+ductal cell differentiation into β cells in mice with reversal of diabetes

“…Because treatment with GE was reported to induce β-cell neogenesis from adult pancreatic ductal cells in vitro (25), and because we observed neogenesis in mixed chimeric late-stage diabetic NOD mice after long-term (8 wk) administration of GE (4), in the present studies, we tested whether long-term administration of GE was able to induce Sox9 + pancreatic ductal cell differentiation into insulinproducing β cells in diabetic C57BL/6 mice, using a Cre-based lineage-tracing construct driven by Sox9 regulatory sequences. The same strain of mice was used as in the studies of Kopp et al (17), but a founder with higher recombination efficiency was used in the present studies.…”

Treatment of autoimmune diabetes in NOD mice by Toll-like receptor 2 tolerance in conjunction with dipeptidyl peptidase 4 inhibition

“…However, safety issues such as teratoma formation and low reprogramming efficiency preclude clinical application of iPS-derived beta cell technology. Other methods increasing compromised beta cell mass comprise diverse pharmacological agents or hormones, including GLP-1 and DPP4 inhibitors [29][30][31][32][33][34][35][36]. In mice treated with Pam3CSK 4 +DA-1229, a beta cell-trophic effect of DPP4 inhibition was demonstrated by increased numbers of proliferating beta cells and small beta cell units.…”