title: Combinatorial insulin secretion dynamics of recombinant hepatic and enteroendocrine cells.
abstract: One of the most promising cell-based therapies for combating insulin-dependent diabetes entails the use of genetically engineered non-β cells that secrete insulin in response to physiologic stimuli. A normal pancreatic β cell secretes insulin in a biphasic manner in response to glucose. The first phase is characterized by a transient stimulation of insulin to rapidly lower the blood glucose levels, which is followed by a second phase of insulin secretion to sustain the lowered blood glucose levels over a longer period of time. Previous studies have demonstrated hepatic and enteroendocrine cells to be appropriate hosts for recombinant insulin expression. Due to different insulin secretion kinetics from these cells, we hypothesized that a combination of the two cell types would mimic the biphasic insulin secretion of normal β cells with higher fidelity than either cell type alone. In this study, insulin secretion experiments were conducted with two hepatic cell lines (HepG2 and H4IIE) transduced with 1 of 3 adenoviruses expressing the insulin transgene and with a stably transfected recombinant intestinal cell line (GLUTag-INS). Insulin secretion was stimulated by exposing the cells to glucose only (hepatic cells), meat hydrolysate only (GLUTag-INS), or to a cocktail of the two secretagogues. It was found experimentally that the recombinant hepatic cells secreted insulin in a more sustained manner, whereas the recombinant intestinal cell line exhibited rapid insulin secretion kinetics upon stimulation. The insulin secretion profiles were computationally combined at different cell ratios to arrive at the combinatorial kinetics. Results indicate that combinations of these two cell types allow for tuning the first and second phase of insulin secretion better than either cell type alone. This work provides the basic framework in understanding the secretion kinetics of the combined system and advances it towards preclinical studies.
authors.affiliation: School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
authors.forename: Kiranmai
authors.initials: K
authors.lastname: Durvasula
authors.forename: Peter M
authors.initials: PM
authors.lastname: Thulé
authors.forename: Athanassios
authors.initials: A
authors.lastname: Sambanis
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002460:Cell Line / Q000187:drug effects / Q000378:metabolism; D018920:Coculture Techniques; D004347:Drug Interactions; D019858:Enteroendocrine Cells / Q000187:drug effects / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D005947:Glucose / Q000494:pharmacology; D056945:Hep G2 Cells / Q000187:drug effects / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D000078790:Insulin Secretion; D008114:Liver Neoplasms, Experimental / Q000473:pathology; D051379:Mice; D011384:Proinsulin / Q000235:genetics; D011492:Protein Hydrolysates / Q000494:pharmacology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012636:Secretory Rate / Q000187:drug effects; D014161:Transduction, Genetic
chemical_list: D007328:Insulin; D011492:Protein Hydrolysates; D011993:Recombinant Fusion Proteins; D011384:Proinsulin; D005947:Glucose
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