Grzegorz Oracz1, Michał Zaród2, Maren Ewers3, Helmut Laumen4, Tomasz Gambin5, Paweł Kamiński6, Iwona Grabowska7, Anna Drożak8, Sebastian Kwiatkowski2, Katarzyna Wertheim-Tysarowska6, Elwira Kołodziejczyk1, Alicja Domaszewicz6, Barbara Dorożko6, Joanna Kosińska9, Stanisław Głuszek10, Dorota Kozieł10, Rafał Płoski9, Jonas Rosendahl11, Heiko Witt3, Jakub Drożak12, Agnieszka Magdalena Rygiel13. 1. Department of Gastroenterology, Hepatology, Feeding Disorders and Pediatrics, The Children's Memorial Health Institute, Warsaw, Poland. 2. Department of Metabolic Regulation, Faculty of Biology, University of Warsaw, Warsaw, Poland. 3. Pediatric Nutritional Medicine & Else Kröner-Fresenius-Centre for Nutritional Medicine (EKFZ), Technical University Munich (TUM), Munich, Germany. 4. Pediatric Nutritional Medicine & Else Kröner-Fresenius-Centre for Nutritional Medicine (EKFZ), Technical University Munich (TUM), Munich, Germany; Department of Internal Medicine I, Martin Luther University, Halle, Germany. 5. Department of Medical Genetics, Institute of Mother and Child, Warsaw, Poland; Institute of Computer Science, Warsaw University of Technology, Warsaw, Poland. 6. Department of Medical Genetics, Institute of Mother and Child, Warsaw, Poland. 7. Department of Cytology, Faculty of Biology, University of Warsaw, Warsaw, Poland. 8. Department of Molecular Plant Physiology, Faculty of Biology, University of Warsaw, Warsaw, Poland. 9. Department of Medical Genetics, Medical University of Warsaw, Warsaw, Poland. 10. Collegium Medicum Jan Kochanowski University, Kielce, Poland. 11. Department of Internal Medicine I, Martin Luther University, Halle, Germany. 12. Department of Metabolic Regulation, Faculty of Biology, University of Warsaw, Warsaw, Poland. Electronic address: jdrozak@biol.uw.edu.pl. 13. Department of Medical Genetics, Institute of Mother and Child, Warsaw, Poland. Electronic address: agnieszka.rygiel@imid.med.pl.
Abstract
PURPOSE: Loss of function variants of the transient receptor potential cation channel, subfamily V, member 6 (TRPV6) have been recently associated with chronic pancreatitis (CP) in Japanese, German and French patients. Here, we investigated the association of TRPV6 variants with CP in independent European cohorts of early-onset CP patients from Poland and Germany. PATIENTS AND METHODS: We enrolled 152 pediatric CP patients (median age 8.6 yrs) with no history of alcohol/smoking abuse and 472 controls from Poland as well as 157 nonalcoholic young CP patients (median age 20 yrs) and 750 controls from Germany. Coding regions of TRPV6 were screened by Sanger and next generation sequencing. Selected, potentially pathogenic TRPV6 variants were expressed in HEK293T cells and TRPV6 activity was analyzed using ratiometric Ca2+ measurements. RESULTS: Overall, we identified 10 novel (3 nonsense and 7 missenses) TRPV6 variants in CP patients. TRPV6 p.V239SfsX53 nonsense variant and the variants showing significant decrease in intracellular Ca2+ concentration in HEK293T cells (p.R174X, p.L576R, p.R342Q), were significantly overrepresented in Polish patients as compared to controls (6/152, 3.9% vs. 0/358, 0%; P = 0,0007). Nonsense TRPV6 variants predicted as loss of function (p.V239SfsX53 and p.R624X) were also significantly overrepresented in German patients (3/157; 2.0% vs 0/750; 0%, P = 0.005). CONCLUSIONS: We showed that TRPV6 loss of function variants are associated with elevated CP risk in early-onset Polish and German patients confirming that TRPV6 is a novel CP susceptibility gene.
PURPOSE: Loss of function variants of the transient receptor potential cation channel, subfamily V, member 6 (TRPV6) have been recently associated with chronic pancreatitis (CP) in Japanese, German and French patients. Here, we investigated the association of TRPV6 variants with CP in independent European cohorts of early-onset CP patients from Poland and Germany. PATIENTS AND METHODS: We enrolled 152 pediatric CP patients (median age 8.6 yrs) with no history of alcohol/smoking abuse and 472 controls from Poland as well as 157 nonalcoholic young CP patients (median age 20 yrs) and 750 controls from Germany. Coding regions of TRPV6 were screened by Sanger and next generation sequencing. Selected, potentially pathogenic TRPV6 variants were expressed in HEK293T cells and TRPV6 activity was analyzed using ratiometric Ca2+ measurements. RESULTS: Overall, we identified 10 novel (3 nonsense and 7 missenses) TRPV6 variants in CP patients. TRPV6 p.V239SfsX53 nonsense variant and the variants showing significant decrease in intracellular Ca2+ concentration in HEK293T cells (p.R174X, p.L576R, p.R342Q), were significantly overrepresented in Polish patients as compared to controls (6/152, 3.9% vs. 0/358, 0%; P = 0,0007). Nonsense TRPV6 variants predicted as loss of function (p.V239SfsX53 and p.R624X) were also significantly overrepresented in German patients (3/157; 2.0% vs 0/750; 0%, P = 0.005). CONCLUSIONS: We showed that TRPV6 loss of function variants are associated with elevated CP risk in early-onset Polish and German patients confirming that TRPV6 is a novel CP susceptibility gene.
Authors: Agnieszka Magdalena Rygiel; Lara Sophie Unger; Franziska Lena Sörgel; Emmanuelle Masson; Ryotaro Matsumoto; Maren Ewers; Jian-Min Chen; Peter Bugert; Louis Buscail; Tomasz Gambin; Grzegorz Oracz; Maria Winiewska-Szajewska; Agnieszka Mianowska; Jarosław Poznanski; Joanna Kosińska; Piotr Stawinski; Rafał Płoski; Dorota Koziel; Stanisław Gluszek; Helmut Laumen; Fredrik Lindgren; J Matthias Löhr; Anna Orekhova; Vinciane Rebours; Jonas Rosendahl; Andrea Párniczky; Péter Hegyi; Akira Sasaki; Fumiya Kataoka; Yu Tanaka; Shin Hamada; Miklós Sahin-Tóth; Eszter Hegyi; Claude Férec; Atsushi Masamune; Heiko Witt Journal: Pancreatology Date: 2022-05-01 Impact factor: 3.977