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Inhibitors of ORAI1 Prevent Cytosolic Calcium-Associated Injury of Human Pancreatic Acinar Cells and Acute Pancreatitis in 3 Mouse Models.

Wen L, Voronina S, Javed MA, Awais M, Szatmary P, Latawiec D, Chvanov M, Collier D, Huang W, Barrett J, Begg M, Stauderman K, Roos J, Grigoryev S, Ramos S, Rogers E, Whitten J, Velicelebi G, Dunn M, Tepikin AV, Criddle DN, Sutton R - Gastroenterology (2015)

Bottom Line: The agents were significantly more effective, in a range of parameters, when given at 1 vs 6 hours after induction of pancreatitis.Cytosolic calcium overload, mediated via ORAI1, contributes to the pathogenesis of acute pancreatitis.ORAI1 inhibitors might be developed for the treatment of patients with pancreatitis.

View Article: PubMed Central - PubMed

Affiliation: Pancreas Biomedical Research Unit, National Institute for Health Research Liverpool, Royal Liverpool University Hospital, Liverpool, United Kingdom; Department of Integrated Traditional and Western Medicine, Sichuan Provincial Pancreatitis Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.

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CM_128 administered from 6 hours after disease induction (late) markedly reduced pancreatic histopathology in TLCS-AP and FAEE-AP. Two models at 6 and 24 hours resulted in substantially progressive increases in (A) edema, (B) inflammation, (C) necrosis, and (D) total histology score, with more marked increase of all scores at 24 hours. Intraperitoneal administration of CM_128 at 20 mg/kg from 6 hours after disease induction significantly reduced edema, but not inflammation, necrosis, or total histology scores at 6 hours (mean ± SEM ≥6 mice/group; *P < .05, control vs 2 models at 6 hours; †P < .05 2 models at 6 vs 24 hours; ‡P < .05, TLCS-AP at 6 h vs TLCS-AP plus CM_128; §P < .05, FAEE-AP at 6 h vs FAEE-AP plus CM_128). (E) Representative images showing normal pancreatic histology, typical histopathology from 2 models at 6 and 24 hours, and typical histopathology from 2 models after treatment with CM_128 administered late after disease induction (H&E; scale bar: 50 μm).
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figS4: CM_128 administered from 6 hours after disease induction (late) markedly reduced pancreatic histopathology in TLCS-AP and FAEE-AP. Two models at 6 and 24 hours resulted in substantially progressive increases in (A) edema, (B) inflammation, (C) necrosis, and (D) total histology score, with more marked increase of all scores at 24 hours. Intraperitoneal administration of CM_128 at 20 mg/kg from 6 hours after disease induction significantly reduced edema, but not inflammation, necrosis, or total histology scores at 6 hours (mean ± SEM ≥6 mice/group; *P < .05, control vs 2 models at 6 hours; †P < .05 2 models at 6 vs 24 hours; ‡P < .05, TLCS-AP at 6 h vs TLCS-AP plus CM_128; §P < .05, FAEE-AP at 6 h vs FAEE-AP plus CM_128). (E) Representative images showing normal pancreatic histology, typical histopathology from 2 models at 6 and 24 hours, and typical histopathology from 2 models after treatment with CM_128 administered late after disease induction (H&E; scale bar: 50 μm).

Mentions: Preliminary in vivo experiments indicated that CM_128 has a significantly longer half-life than GSK-7975A, and is suitable for intraperitoneal dosing every 12 hours to achieve sustained blood levels with more than 99% bound (free fraction in murine plasma, 0.33%; when added to human plasma, 0.16%). Because our work with high-dose GSK-7975A showed greater efficacy in vivo than with low-dose GSK-7975A, and in vitro data obtained with CM_128 did not suggest loss of efficacy at high concentrations (10 μmol/L), we administered 20 mg/kg CM_128 every 12 hours to test the efficacy of this agent in TLCS-AP and FAEE-AP. We also determined the relative efficacy of CM_128 administered either 1 or 6 hours after induction of either model of acute pancreatitis. CM_128 begun 1 hour after disease induction significantly reduced all parameters of both TLCS-AP and FAEE-AP, including all local and systemic biochemical, immunologic, and histopathologic measures (Figures 6 and 7). CM_128 begun 6 hours after disease induction was less effective across a broad range of parameters (Figures 6 and 7), significantly so for IL6 (TLCS-AP), pancreatic MPO (FAEE-AP), and lung MPO (TLCS-AP), although significant reductions still were seen in amylase (TLCS-AP and FAEE-AP), lung MPO (TLCS-AP), edema (TLCS-AP and FAEE-AP), inflammation (FAEE-AP), necrosis (FAEE-AP), and total histopathology score (FAEE-AP). To determine the extent to which disease was established at 6 hours after disease induction, and the effect of CM_128 begun at that time, all parameters were assessed at 6 hours and compared with values at 24 hours. These data showed that by 24 hours there was no significant improvement of parameters as measured at 6 hours as a result of CM_128 administration begun at 6 hours (Supplementary Figures 3 and 4), confirming delay in therapy to be disadvantageous; although CM_128 appeared to prevent these parameters from increasing. To further explore the effect of delay in dosing, the effect of high-dose GSK-7975A on disease responses also was tested at 1 and 6 hours after induction. Similar to CM_128, GSK-7975A begun 6 hours after disease induction was less effective across a broad range of parameters (Supplementary Figures 5 and 6), significantly so for amylase (TLCS-AP and FAEE-AP), IL6 (TLCS-AP), edema (TLCS-AP and FAEE-AP), inflammatory infiltrate (TLCS-AP), and total histopathology score (TLCS-AP and FAEE-AP).


Inhibitors of ORAI1 Prevent Cytosolic Calcium-Associated Injury of Human Pancreatic Acinar Cells and Acute Pancreatitis in 3 Mouse Models.

Wen L, Voronina S, Javed MA, Awais M, Szatmary P, Latawiec D, Chvanov M, Collier D, Huang W, Barrett J, Begg M, Stauderman K, Roos J, Grigoryev S, Ramos S, Rogers E, Whitten J, Velicelebi G, Dunn M, Tepikin AV, Criddle DN, Sutton R - Gastroenterology (2015)

CM_128 administered from 6 hours after disease induction (late) markedly reduced pancreatic histopathology in TLCS-AP and FAEE-AP. Two models at 6 and 24 hours resulted in substantially progressive increases in (A) edema, (B) inflammation, (C) necrosis, and (D) total histology score, with more marked increase of all scores at 24 hours. Intraperitoneal administration of CM_128 at 20 mg/kg from 6 hours after disease induction significantly reduced edema, but not inflammation, necrosis, or total histology scores at 6 hours (mean ± SEM ≥6 mice/group; *P < .05, control vs 2 models at 6 hours; †P < .05 2 models at 6 vs 24 hours; ‡P < .05, TLCS-AP at 6 h vs TLCS-AP plus CM_128; §P < .05, FAEE-AP at 6 h vs FAEE-AP plus CM_128). (E) Representative images showing normal pancreatic histology, typical histopathology from 2 models at 6 and 24 hours, and typical histopathology from 2 models after treatment with CM_128 administered late after disease induction (H&E; scale bar: 50 μm).
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Related In: Results  -  Collection

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figS4: CM_128 administered from 6 hours after disease induction (late) markedly reduced pancreatic histopathology in TLCS-AP and FAEE-AP. Two models at 6 and 24 hours resulted in substantially progressive increases in (A) edema, (B) inflammation, (C) necrosis, and (D) total histology score, with more marked increase of all scores at 24 hours. Intraperitoneal administration of CM_128 at 20 mg/kg from 6 hours after disease induction significantly reduced edema, but not inflammation, necrosis, or total histology scores at 6 hours (mean ± SEM ≥6 mice/group; *P < .05, control vs 2 models at 6 hours; †P < .05 2 models at 6 vs 24 hours; ‡P < .05, TLCS-AP at 6 h vs TLCS-AP plus CM_128; §P < .05, FAEE-AP at 6 h vs FAEE-AP plus CM_128). (E) Representative images showing normal pancreatic histology, typical histopathology from 2 models at 6 and 24 hours, and typical histopathology from 2 models after treatment with CM_128 administered late after disease induction (H&E; scale bar: 50 μm).
Mentions: Preliminary in vivo experiments indicated that CM_128 has a significantly longer half-life than GSK-7975A, and is suitable for intraperitoneal dosing every 12 hours to achieve sustained blood levels with more than 99% bound (free fraction in murine plasma, 0.33%; when added to human plasma, 0.16%). Because our work with high-dose GSK-7975A showed greater efficacy in vivo than with low-dose GSK-7975A, and in vitro data obtained with CM_128 did not suggest loss of efficacy at high concentrations (10 μmol/L), we administered 20 mg/kg CM_128 every 12 hours to test the efficacy of this agent in TLCS-AP and FAEE-AP. We also determined the relative efficacy of CM_128 administered either 1 or 6 hours after induction of either model of acute pancreatitis. CM_128 begun 1 hour after disease induction significantly reduced all parameters of both TLCS-AP and FAEE-AP, including all local and systemic biochemical, immunologic, and histopathologic measures (Figures 6 and 7). CM_128 begun 6 hours after disease induction was less effective across a broad range of parameters (Figures 6 and 7), significantly so for IL6 (TLCS-AP), pancreatic MPO (FAEE-AP), and lung MPO (TLCS-AP), although significant reductions still were seen in amylase (TLCS-AP and FAEE-AP), lung MPO (TLCS-AP), edema (TLCS-AP and FAEE-AP), inflammation (FAEE-AP), necrosis (FAEE-AP), and total histopathology score (FAEE-AP). To determine the extent to which disease was established at 6 hours after disease induction, and the effect of CM_128 begun at that time, all parameters were assessed at 6 hours and compared with values at 24 hours. These data showed that by 24 hours there was no significant improvement of parameters as measured at 6 hours as a result of CM_128 administration begun at 6 hours (Supplementary Figures 3 and 4), confirming delay in therapy to be disadvantageous; although CM_128 appeared to prevent these parameters from increasing. To further explore the effect of delay in dosing, the effect of high-dose GSK-7975A on disease responses also was tested at 1 and 6 hours after induction. Similar to CM_128, GSK-7975A begun 6 hours after disease induction was less effective across a broad range of parameters (Supplementary Figures 5 and 6), significantly so for amylase (TLCS-AP and FAEE-AP), IL6 (TLCS-AP), edema (TLCS-AP and FAEE-AP), inflammatory infiltrate (TLCS-AP), and total histopathology score (TLCS-AP and FAEE-AP).

Bottom Line: The agents were significantly more effective, in a range of parameters, when given at 1 vs 6 hours after induction of pancreatitis.Cytosolic calcium overload, mediated via ORAI1, contributes to the pathogenesis of acute pancreatitis.ORAI1 inhibitors might be developed for the treatment of patients with pancreatitis.

View Article: PubMed Central - PubMed

Affiliation: Pancreas Biomedical Research Unit, National Institute for Health Research Liverpool, Royal Liverpool University Hospital, Liverpool, United Kingdom; Department of Integrated Traditional and Western Medicine, Sichuan Provincial Pancreatitis Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.

Show MeSH
Related in: MedlinePlus