Glucagonlike Peptide 2 Analogue Teduglutide

Abstract

IMPORTANCE Short bowel syndrome occurs when a shortened intestine cannot absorb sufficient nutrients or fluids. Teduglutide is a recombinant analogue of human glucagonlike peptide 2 that reduces dependence on parenteral nutrition in patients with short bowel syndrome by promoting enterocytic proliferation, increasing the absorptive surface area. However, enterocyte function depends not only on the number of cells that are present but also on differentiated features that facilitate nutrient absorption and digestion. OBJECTIVE To test the hypothesis that teduglutide impairs human intestinal epithelial differentiation. DESIGN AND SETTING We investigated the effects of teduglutide in the modulation of proliferation and differentiation in human Caco-2 intestinal epithelial cells at a basic science laboratory. This was an in vitro study using Caco-2 cells, a human-derived intestinal epithelial cell line commonly used to model enterocytic biology. EXPOSURE Cells were exposed to teduglutide or vehicle control. MAINOUTCOMESAND MEASURES We analyzed the cell cycle by bromodeoxyuridine incorporation or propidium iodide staining and flow cytometry and measured cell proliferation by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. We used quantitative reverse transcription–polymerase chain reaction to assay the expression of the enterocytic differentiation markers villin, sucrase-isomaltase, glucose transporter 2 (GLUT2), and dipeptidyl peptidase 4 (DPP-4), as well as that of the putative differentiation signals schlafen 12 (SLFN12) and caudal-related homeobox intestine-specific transcription factor (Cdx2). Villin promoter activity was measured by a luciferase-based assay. RESULTS The MTS assay demonstrated that teduglutide increased cell numbers by a mean (SD) of 10% (2%) over untreated controls at a maximal 500nM (n = 6, P < .05). Teduglutide increased bromodeoxyuridine-positive cells vs untreated controls by a mean (SD) of 19.4% (2.3%) vs 12.0% (0.8%) (n = 6, P < .05) and increased the S-phase fraction by flow cytometric analysis. Teduglutide reduced the mean (SD) expression of villin by 29% (6%), Cdx2 by 31% (10%), DPP-4 by 15% (6%), GLUT2 by 40% (11%), SLFN12 by 61% (14%), and sucrase-isomaltase by 28% (8%) (n = 6, P < .05 for all). CONCLUSIONS AND RELEVANCE Teduglutide increased Caco-2 proliferation but tended to inhibit intestinal epithelial differentiation. The effects of mitogenic stimulation with teduglutide in patients with short bowel syndrome might be greater if the more numerous teduglutide-treated cells could be stimulated toward a more fully differentiated phenotype.

Schlafen 12 Interaction with SerpinB12 and Deubiquitylases Drives Human Enterocyte Differentiation

“…Most attention has focused on increasing mucosal mass in short gut with enterocytic mitogens like glutamine and teduglutide that may actually decrease enterocytic differentiation [61, 62], and on decreasing available intestinal mucosal mass by Roux-en-Y intestinal bypass in morbid obesity. Our results raise the possibility that enterocytic differentiation can be manipulated directly independently of proliferation to enhance or reduce enterocyte nutrient absorption and digestion.…”

Schlafen 12 mediates the effects of butyrate and repetitive mechanical deformation on intestinal epithelial differentiation in human Caco-2 intestinal epithelial cells

“…Both glutamine and teduglutide may be used in this fashion. However, while glutamine [1,2] and teduglutide [3] stimulate intestinal epithelial proliferation, speeding proliferation may also decrease the time available for enterocyte maturation, reducing the individual digestive and absorptive capacity of individual enterocytes. An ideal approach would therefore target both enterocyte proliferation and differentiation.…”

Schlafen 3 Mediates the Differentiating Effects of Cdx2 in Rat IEC-Cdx2L1 Enterocytes

“…We have previously traced in vitro influences on intestinal epithelial differentiation by luminal contents such as butyrate [29–31] and glutamine [32], growth factors such as TGF-β [33] or somatostatin [34], chemical factors like pH [35], and even physical forces like repetitive deformation during peristalsis or villous motility [36, 37]. In short gut syndrome, when there is inadequate mucosal capacity to maintain nutrition [2], attention has chiefly focused on the use of mitogens such as teduglutide to stimulate enterocytic proliferation to increase mucosal mass, but such mitogenic stimuli may actually reduce differentiation [38]. Conversely, malignancy represents an obvious loss of differentiation while proliferation increases.…”