Genetic and functional analysis of chymotrypsin-like protease (CTRL) in chronic pancreatitis.

CTRL Chronic pancreatitis Chymotrypsin-like protease Genetics

Journal

Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.]
ISSN: 1424-3911
Titre abrégé: Pancreatology
Pays: Switzerland
ID NLM: 100966936

Informations de publication

Date de publication:
Dec 2023
Historique:
received: 22 09 2023
revised: 30 10 2023
accepted: 01 11 2023
pubmed: 11 11 2023
medline: 11 11 2023
entrez: 10 11 2023
Statut: ppublish

Résumé

Genetic predisposition is crucial in the pathogenesis of early-onset chronic pancreatitis (CP). So far, several genetic alterations have been identified as risk factors, predominantly in genes encoding digestive enzymes. However, many early-onset CP cases have no identified underlying cause. Chymotrypsins are a family of serine proteases that can cleave trypsinogen and lead to its degradation. Because genetic alterations in the chymotrypsins CTRC, CTRB1, and CTRB2 are associated with CP, we genetically and functionally investigated chymotrypsin-like protease (CTRL) as a potential risk factor. We screened 1005 non-alcoholic CP patients and 1594 controls for CTRL variants by exome sequencing. We performed Western blots and activity assays to analyse secretion and proteolytic activity. We measured BiP mRNA expression to investigate the potential impact of identified alterations on endoplasmic reticulum (ER) stress. We identified 13 heterozygous non-synonymous CTRL variants: five exclusively in patients and three only in controls. Functionality was unchanged in 6/13 variants. Four alterations showed normal secretion but reduced (p.G20S, p.G56S, p.G61S) or abolished (p.S208F) activity. Another three variants (p.C201Y, p.G215R and p.C220G) were not secreted and already showed reduced or no activity intracellularly. However, intracellular retention did not lead to ER stress. We identified several CTRL variants, some showing potent effects on protease function and secretion. We observed these effects in variants found in patients and controls, and CTRL loss-of-function variants were not significantly more common in patients than controls. Therefore, CTRL is unlikely to play a relevant role in the development of CP.

Sections du résumé

BACKGROUND BACKGROUND
Genetic predisposition is crucial in the pathogenesis of early-onset chronic pancreatitis (CP). So far, several genetic alterations have been identified as risk factors, predominantly in genes encoding digestive enzymes. However, many early-onset CP cases have no identified underlying cause. Chymotrypsins are a family of serine proteases that can cleave trypsinogen and lead to its degradation. Because genetic alterations in the chymotrypsins CTRC, CTRB1, and CTRB2 are associated with CP, we genetically and functionally investigated chymotrypsin-like protease (CTRL) as a potential risk factor.
METHODS METHODS
We screened 1005 non-alcoholic CP patients and 1594 controls for CTRL variants by exome sequencing. We performed Western blots and activity assays to analyse secretion and proteolytic activity. We measured BiP mRNA expression to investigate the potential impact of identified alterations on endoplasmic reticulum (ER) stress.
RESULTS RESULTS
We identified 13 heterozygous non-synonymous CTRL variants: five exclusively in patients and three only in controls. Functionality was unchanged in 6/13 variants. Four alterations showed normal secretion but reduced (p.G20S, p.G56S, p.G61S) or abolished (p.S208F) activity. Another three variants (p.C201Y, p.G215R and p.C220G) were not secreted and already showed reduced or no activity intracellularly. However, intracellular retention did not lead to ER stress.
CONCLUSION CONCLUSIONS
We identified several CTRL variants, some showing potent effects on protease function and secretion. We observed these effects in variants found in patients and controls, and CTRL loss-of-function variants were not significantly more common in patients than controls. Therefore, CTRL is unlikely to play a relevant role in the development of CP.

Identifiants

pubmed: 37949771
pii: S1424-3903(23)01839-2
doi: 10.1016/j.pan.2023.11.002
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

957-963

Informations de copyright

Copyright © 2023 IAP and EPC. Published by Elsevier B.V. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of competing interest The authors report no conflicts of interest.

Auteurs

Katharina Eiseler (K)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany.

Lea Neppl (L)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany.

Andreas W Schmidt (AW)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany; Department of Internal Medicine I, Martin Luther University (MLU), Halle (Saale), Germany; Institute of Medical Genetics and Applied Genomics, University of Tübingen, Tübingen, Germany.

Beate Rauscher (B)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany.

Maren Ewers (M)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany.

Emmanuelle Masson (E)

Univ Brest, Inserm, EFS, UMR 1078, GGB, F-29200, Brest, France; Service de Génétique Médicale et de Biologie de la Reproduction, CHRU Brest, F-29200, Brest, France.

Jian-Min Chen (JM)

Univ Brest, Inserm, EFS, UMR 1078, GGB, F-29200, Brest, France.

Claude Férec (C)

Univ Brest, Inserm, EFS, UMR 1078, GGB, F-29200, Brest, France.

Vinciane Rebours (V)

Pancreatology and Digestive Oncology Department, Beaujon Hospital, APHP - Clichy, Université Paris Cité, Paris, France.

Tassos Grammatikopoulos (T)

Paediatric Liver, GI & Nutrition Centre and MowatLabs, King's College Hospital NHS Foundation Trust, London, UK; Institute of Liver Studies, King's College London, London, UK.

Pierre Foskett (P)

Institute of Liver Studies, King's College London, London, UK.

William Greenhalf (W)

Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK.

Christopher Halloran (C)

Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, UK.

John Neoptolemos (J)

Department of Surgery, University of Heidelberg, Heidelberg, Germany.

Tobias B Haack (TB)

Institute of Medical Genetics and Applied Genomics, University of Tübingen, Tübingen, Germany.

Stephan Ossowski (S)

Institute of Medical Genetics and Applied Genomics, University of Tübingen, Tübingen, Germany.

Marc Sturm (M)

Institute of Medical Genetics and Applied Genomics, University of Tübingen, Tübingen, Germany.

Jonas Rosendahl (J)

Department of Internal Medicine I, Martin Luther University (MLU), Halle (Saale), Germany.

Helmut Laumen (H)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany; Department of Internal Medicine I, Martin Luther University (MLU), Halle (Saale), Germany.

Heiko Witt (H)

Paediatric Nutritional Medicine, Else Kröner Fresenius Center for Nutritional Medicine (EKFZ), Technical University of Munich (TUM), Freising, Germany. Electronic address: heiko.witt@tum.de.

Classifications MeSH