TY - JOUR
T1 - Hyperglycemia Increases Interstitial Cells of Cajal via MAPK1 and MAPK3 Signaling to ETV1 and KIT, Leading to Rapid Gastric Emptying
AU - Hayashi, Yujiro
AU - Toyomasu, Yoshitaka
AU - Saravanaperumal, Siva Arumugam
AU - Bardsley, Michael R.
AU - Smestad, John A.
AU - Lorincz, Andrea
AU - Eisenman, Seth T.
AU - Cipriani, Gianluca
AU - Nelson Holte, Molly H.
AU - Al Khazal, Fatimah J.
AU - Syed, Sabriya A.
AU - Gajdos, Gabriella B.
AU - Choi, Kyoung Moo
AU - Stoltz, Gary J.
AU - Miller, Katie E.
AU - Kendrick, Michael L.
AU - Rubin, Brian P.
AU - Gibbons, Simon J.
AU - Bharucha, Adil E.
AU - Linden, David R.
AU - Maher, Louis James
AU - Farrugia, Gianrico
AU - Ordog, Tamas
N1 - Publisher Copyright:
© 2017 AGA Institute
PY - 2017/8
Y1 - 2017/8
N2 - Background & Aims Depletion of interstitial cells of Cajal (ICCs) is common in diabetic gastroparesis. However, in approximately 20% of patients with diabetes, gastric emptying (GE) is accelerated. GE also occurs faster in obese individuals, and is associated with increased blood levels of glucose in patients with type 2 diabetes. To understand the fate of ICCs in hyperinsulinemic, hyperglycemic states characterized by rapid GE, we studied mice with mutation of the leptin receptor (Leprdb/db), which in our colony had accelerated GE. We also investigated hyperglycemia-induced signaling in the ICC lineage and ICC dependence on glucose oxidative metabolism in mice with disruption of the succinate dehydrogenase complex, subunit C gene (Sdhc). Methods Mice were given breath tests to analyze GE of solids. ICCs were studied by flow cytometry, intracellular electrophysiology, isometric contractility measurement, reverse-transcription polymerase chain reaction, immunoblot, immunohistochemistry, enzyme-linked immunosorbent assays, and metabolite assays; cells and tissues were manipulated pharmacologically and by RNA interference. Viable cell counts, proliferation, and apoptosis were determined by methyltetrazolium, Ki-67, proliferating cell nuclear antigen, bromodeoxyuridine, and caspase–Glo 3/7 assays. Sdhc was disrupted in 2 different strains of mice via cre recombinase. Results In obese, hyperglycemic, hyperinsulinemic female Leprdb/db mice, GE was accelerated and gastric ICC and phasic cholinergic responses were increased. Female KitK641E/+ mice, which have genetically induced hyperplasia of ICCs, also had accelerated GE. In isolated cells of the ICC lineage and gastric organotypic cultures, hyperglycemia stimulated proliferation by mitogen-activated protein kinase 1 (MAPK1)- and MAPK3-dependent stabilization of ets variant 1—a master transcription factor for ICCs—and consequent up-regulation of v-kit Hardy–Zuckerman 4 feline sarcoma viral oncogene homolog (KIT) receptor tyrosine kinase. Opposite changes occurred in mice with disruption of Sdhc. Conclusions Hyperglycemia increases ICCs via oxidative metabolism-dependent, MAPK1- and MAPK3-mediated stabilization of ets variant 1 and increased expression of KIT, causing rapid GE. Increases in ICCs might contribute to the acceleration in GE observed in some patients with diabetes.
AB - Background & Aims Depletion of interstitial cells of Cajal (ICCs) is common in diabetic gastroparesis. However, in approximately 20% of patients with diabetes, gastric emptying (GE) is accelerated. GE also occurs faster in obese individuals, and is associated with increased blood levels of glucose in patients with type 2 diabetes. To understand the fate of ICCs in hyperinsulinemic, hyperglycemic states characterized by rapid GE, we studied mice with mutation of the leptin receptor (Leprdb/db), which in our colony had accelerated GE. We also investigated hyperglycemia-induced signaling in the ICC lineage and ICC dependence on glucose oxidative metabolism in mice with disruption of the succinate dehydrogenase complex, subunit C gene (Sdhc). Methods Mice were given breath tests to analyze GE of solids. ICCs were studied by flow cytometry, intracellular electrophysiology, isometric contractility measurement, reverse-transcription polymerase chain reaction, immunoblot, immunohistochemistry, enzyme-linked immunosorbent assays, and metabolite assays; cells and tissues were manipulated pharmacologically and by RNA interference. Viable cell counts, proliferation, and apoptosis were determined by methyltetrazolium, Ki-67, proliferating cell nuclear antigen, bromodeoxyuridine, and caspase–Glo 3/7 assays. Sdhc was disrupted in 2 different strains of mice via cre recombinase. Results In obese, hyperglycemic, hyperinsulinemic female Leprdb/db mice, GE was accelerated and gastric ICC and phasic cholinergic responses were increased. Female KitK641E/+ mice, which have genetically induced hyperplasia of ICCs, also had accelerated GE. In isolated cells of the ICC lineage and gastric organotypic cultures, hyperglycemia stimulated proliferation by mitogen-activated protein kinase 1 (MAPK1)- and MAPK3-dependent stabilization of ets variant 1—a master transcription factor for ICCs—and consequent up-regulation of v-kit Hardy–Zuckerman 4 feline sarcoma viral oncogene homolog (KIT) receptor tyrosine kinase. Opposite changes occurred in mice with disruption of Sdhc. Conclusions Hyperglycemia increases ICCs via oxidative metabolism-dependent, MAPK1- and MAPK3-mediated stabilization of ets variant 1 and increased expression of KIT, causing rapid GE. Increases in ICCs might contribute to the acceleration in GE observed in some patients with diabetes.
KW - ERK
KW - Gastrointestinal Motility
KW - Mesenchymal Cells
KW - Signal Transduction
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U2 - 10.1053/j.gastro.2017.04.020
DO - 10.1053/j.gastro.2017.04.020
M3 - Article
C2 - 28438610
AN - SCOPUS:85024927473
SN - 0016-5085
VL - 153
SP - 521-535.e20
JO - Gastroenterology
JF - Gastroenterology
IS - 2
ER -