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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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GSK-7975A inhibits CRAC entry (Fura-2 340:380 normalized at 700 s). (A) Changes in mouse pancreatic acinar [Ca2+]C induced by CCK (1 nmol/L) with external physiological [Ca2+] (1.8 mmol/L) applied, showing the effect of 15 μmol/L GSK-7975A from 700 seconds, expanded (≥79 cells/group). Changes in mouse pancreatic acinar [Ca2+]C induced by (B) TLCS (500 μmol/L) and (C) CCK (1 nmol/L), showing effects of 50 and 100 μmol/L GSK-7975A from 700 seconds, expanded. (D and E) Mean (±SEM) [Ca2+]C at 700, 1200, and 2000 seconds from panels B and C, showing a marked reduction with 50 μmol/L GSK-7975A, but not 100 μmol/L GSK-7975A (≥27 cells/group; *P < .001, toxin vs toxin plus GSK-7975A at 1200 s; †P < .001, at 2000 s).
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figS1: GSK-7975A inhibits CRAC entry (Fura-2 340:380 normalized at 700 s). (A) Changes in mouse pancreatic acinar [Ca2+]C induced by CCK (1 nmol/L) with external physiological [Ca2+] (1.8 mmol/L) applied, showing the effect of 15 μmol/L GSK-7975A from 700 seconds, expanded (≥79 cells/group). Changes in mouse pancreatic acinar [Ca2+]C induced by (B) TLCS (500 μmol/L) and (C) CCK (1 nmol/L), showing effects of 50 and 100 μmol/L GSK-7975A from 700 seconds, expanded. (D and E) Mean (±SEM) [Ca2+]C at 700, 1200, and 2000 seconds from panels B and C, showing a marked reduction with 50 μmol/L GSK-7975A, but not 100 μmol/L GSK-7975A (≥27 cells/group; *P < .001, toxin vs toxin plus GSK-7975A at 1200 s; †P < .001, at 2000 s).

Mentions: Isolated murine pancreatic acinar cells maintained in 5 mmol/L external Ca2+ were perfused with TLCS (500 μmol/L) or supramaximal CCK (1 nmol/L) to induce sustained increases of [Ca2+]C dependent on SOCE.1,3,6,10 Once a stable plateau in [Ca2+]C had formed, a range of fixed concentrations (0–100 μmol/L) of GSK-7975A were added. Increasing concentrations of GSK-7975A decreased the [Ca2+]C plateau progressively and increasingly rapidly (Figure 2A–D). With TLCS, suppression of [Ca2+]C toward the initial baseline approached 80% using 30 μmol/L GSK-7975A; with CCK, more than 95% using 15 μmol/L GSK-7975A, an effect also seen when cells were maintained in 1.8 mmol/L external Ca2+ (Supplementary Figure 1A). At 100 μmol/L GSK-7975A, but not at 50 μmol/L GSK-7975A, there was a loss of effect through an unknown mechanism (Supplementary Figure 1B–E). Necrotic cell death pathway activation was reduced markedly in murine pancreatic acinar cells by GSK-7975A (Figure 2E).


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)

GSK-7975A inhibits CRAC entry (Fura-2 340:380 normalized at 700 s). (A) Changes in mouse pancreatic acinar [Ca2+]C induced by CCK (1 nmol/L) with external physiological [Ca2+] (1.8 mmol/L) applied, showing the effect of 15 μmol/L GSK-7975A from 700 seconds, expanded (≥79 cells/group). Changes in mouse pancreatic acinar [Ca2+]C induced by (B) TLCS (500 μmol/L) and (C) CCK (1 nmol/L), showing effects of 50 and 100 μmol/L GSK-7975A from 700 seconds, expanded. (D and E) Mean (±SEM) [Ca2+]C at 700, 1200, and 2000 seconds from panels B and C, showing a marked reduction with 50 μmol/L GSK-7975A, but not 100 μmol/L GSK-7975A (≥27 cells/group; *P < .001, toxin vs toxin plus GSK-7975A at 1200 s; †P < .001, at 2000 s).
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Related In: Results  -  Collection

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figS1: GSK-7975A inhibits CRAC entry (Fura-2 340:380 normalized at 700 s). (A) Changes in mouse pancreatic acinar [Ca2+]C induced by CCK (1 nmol/L) with external physiological [Ca2+] (1.8 mmol/L) applied, showing the effect of 15 μmol/L GSK-7975A from 700 seconds, expanded (≥79 cells/group). Changes in mouse pancreatic acinar [Ca2+]C induced by (B) TLCS (500 μmol/L) and (C) CCK (1 nmol/L), showing effects of 50 and 100 μmol/L GSK-7975A from 700 seconds, expanded. (D and E) Mean (±SEM) [Ca2+]C at 700, 1200, and 2000 seconds from panels B and C, showing a marked reduction with 50 μmol/L GSK-7975A, but not 100 μmol/L GSK-7975A (≥27 cells/group; *P < .001, toxin vs toxin plus GSK-7975A at 1200 s; †P < .001, at 2000 s).
Mentions: Isolated murine pancreatic acinar cells maintained in 5 mmol/L external Ca2+ were perfused with TLCS (500 μmol/L) or supramaximal CCK (1 nmol/L) to induce sustained increases of [Ca2+]C dependent on SOCE.1,3,6,10 Once a stable plateau in [Ca2+]C had formed, a range of fixed concentrations (0–100 μmol/L) of GSK-7975A were added. Increasing concentrations of GSK-7975A decreased the [Ca2+]C plateau progressively and increasingly rapidly (Figure 2A–D). With TLCS, suppression of [Ca2+]C toward the initial baseline approached 80% using 30 μmol/L GSK-7975A; with CCK, more than 95% using 15 μmol/L GSK-7975A, an effect also seen when cells were maintained in 1.8 mmol/L external Ca2+ (Supplementary Figure 1A). At 100 μmol/L GSK-7975A, but not at 50 μmol/L GSK-7975A, there was a loss of effect through an unknown mechanism (Supplementary Figure 1B–E). Necrotic cell death pathway activation was reduced markedly in murine pancreatic acinar cells by GSK-7975A (Figure 2E).

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