2. Academic Validation
  3. Variants That Affect Function of Calcium Channel TRPV6 Are Associated With Early-Onset Chronic Pancreatitis

Variants That Affect Function of Calcium Channel TRPV6 Are Associated With Early-Onset Chronic Pancreatitis

  • Gastroenterology. 2020 May;158(6):1626-1641.e8. doi: 10.1053/j.gastro.2020.01.005.
Atsushi Masamune 1,  Hiroshi Kotani 2,  Franziska Lena Sörgel 3,  Jian-Min Chen 4,  Shin Hamada 5,  Reiko Sakaguchi 6,  Emmanuelle Masson 7,  Eriko Nakano 5,  Yoichi Kakuta 5,  Tetsuya Niihori 8,  Ryo Funayama 9,  Matsuyuki Shirota 9,  Tatsuya Hirano 2,  Tetsuya Kawamoto 2,  Atsuki Hosokoshi 2,  Kiyoshi Kume 5,  Lara Unger 3,  Maren Ewers 3,  Helmut Laumen 3,  Peter Bugert 10,  Masayuki X Mori 2,  Volodymyr Tsvilovskyy 11,  Petra Weißgerber 12,  Ulrich Kriebs 11,  Claudia Fecher-Trost 12,  Marc Freichel 11,  Kalliope N Diakopoulos 13,  Alexandra Berninger 13,  Marina Lesina 13,  Kentaro Ishii 14,  Takao Itoi 14,  Tsukasa Ikeura 15,  Kazuichi Okazaki 15,  Tom Kaune 16,  Jonas Rosendahl 16,  Masao Nagasaki 17,  Yasuhito Uezono 18,  Hana Algül 13,  Keiko Nakayama 9,  Yoichi Matsubara 19,  Yoko Aoki 8,  Claude Férec 7,  Yasuo Mori 2,  Heiko Witt 3,  Tooru Shimosegawa 5
Affiliations

Affiliations

  • 1 Division of Gastroenterology, Tohoku University Graduate School of Medicine, Sendai, Japan. Electronic address: [email protected].
  • 2 Laboratory of Molecular Biology, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
  • 3 Else Kröner-Fresenius-Zentrum für Ernährungsmedizin, Paediatric Nutritional Medicine, Technische Universität München, Freising, Germany.
  • 4 Inserm, Univ Brest, EFS, UMR 1078, GGB, Brest, France.
  • 5 Division of Gastroenterology, Tohoku University Graduate School of Medicine, Sendai, Japan.
  • 6 Laboratory of Molecular Biology, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan; Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan.
  • 7 Inserm, Univ Brest, EFS, UMR 1078, GGB, Brest, France; CHU Brest, Service de Génétique, Brest, France.
  • 8 Department of Medical Genetics, Tohoku University Graduate School of Medicine, Sendai, Japan.
  • 9 Division of Cell Proliferation, United Centers for Advanced Research and Translational Medicine, Tohoku University Graduate School of Medicine, Sendai, Japan.
  • 10 Institute of Transfusion Medicine and Immunology, Medical Faculty Mannheim, Heidelberg University, German Red Cross Blood Service of Baden-Württemberg-Hessen, Mannheim, Germany.
  • 11 Pharmakologisches Institut, Universität Heidelberg, Heidelberg, Germany; German Center for Cardiovascular Research, partner site Heidelberg, Germany.
  • 12 Experimentelle und Klinische Pharmakologie und Toxikologie, Universität des Saarlandes, Homburg, Germany.
  • 13 Mildred Scheel Chair of Tumor Metabolism and Comprehensive Cancer Center Munich at the Klinikum rechts der Isar, Technical University of Munich, Munich, Germany.
  • 14 Department of Gastroenterology and Hepatology, Tokyo Medical University, Tokyo, Japan.
  • 15 Department of Gastroenterology and Hepatology, Kansai Medical University, Hirakata, Japan.
  • 16 Department of Internal Medicine I, Martin Luther University, Halle (Saale), Germany.
  • 17 Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan.
  • 18 Cancer Pathophysiology Division, National Cancer Center Research Institute, Tokyo, Japan.
  • 19 National Center for Child Health and Development, Tokyo, Japan.
PMID: 31930989 DOI: 10.1053/j.gastro.2020.01.005
Abstract

Background & aims: Changes in pancreatic calcium levels affect secretion and might be involved in development of chronic pancreatitis (CP). We investigated the association of CP with the transient receptor potential cation channel subfamily V member 6 gene (TRPV6), which encodes a CA2+-selective ion channel, in an international cohort of patients and in mice.

Methods: We performed whole-exome DNA Sequencing from a patient with idiopathic CP and from his parents, who did not have CP. We validated our findings by Sequencing DNA from 300 patients with CP (not associated with alcohol consumption) and 1070 persons from the general population in Japan (control individuals). In replication studies, we sequenced DNA from patients with early-onset CP (20 years or younger) not associated with alcohol consumption from France (n = 470) and Germany (n = 410). We expressed TRPV6 variants in HEK293 cells and measured their activity using CA2+ imaging assays. CP was induced by repeated injections of cerulein in TRPV6mut/mut mice.

Results: We identified the variants c.629C>T (p.A210V) and c.970G>A (p.D324N) in TRPV6 in the index patient. Variants that affected function of the TRPV6 product were found in 13 of 300 patients (4.3%) and 1 of 1070 control individuals (0.1%) from Japan (odds ratio [OR], 48.4; 95% confidence interval [CI], 6.3-371.7; P = 2.4 × 10-8). Twelve of 124 patients (9.7%) with early-onset CP had such variants. In the replication set from Europe, 18 patients with CP (2.0%) carried variants that affected the function of the TRPV6 product compared with 0 control individuals (P = 6.2 × 10-8). Variants that did not affect the function of the TRPV6 product (p.I223T and p.D324N) were overrepresented in Japanese patients vs control individuals (OR, 10.9; 95% CI, 4.5-25.9; P = 7.4 × 10-9 for p.I223T and P = .01 for p.D324N), whereas the p.L299Q was overrepresented in European patients vs control individuals (OR, 3.0; 95% CI, 1.9-4.8; P = 1.2 × 10-5). TRPV6mut/mut mice given cerulein developed more severe pancreatitis than control mice, as shown by increased levels of pancreatic Enzymes, histologic alterations, and pancreatic fibrosis.

Conclusions: We found that patients with early-onset CP not associated with alcohol consumption carry variants in TRPV6 that affect the function of its product, perhaps by altering CA2+ balance in pancreatic cells. TRPV6 regulates CA2+ homeostasis and pancreatic inflammation.

Keywords

Genetics; Next-generation Sequencing; Transient Receptor Potential; Whole Exome Sequencing.

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