Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis

Johannes R. Hov; Verena Keitel; Jon K. Laerdahl; Lina Spomer; Eva Ellinghaus; Abdou ElSharawy; Espen Melum; Kirsten M. Boberg; Thomas Manke; Tobias Balschun; Christoph Schramm; Annika Bergquist; Tobias Weismueller; Daniel Gotthardt; Christian Rust; Liesbet Henckaerts; Clive M. Onnie; Rinse K. Weersma; Martina Sterneck; Andreas Teufel; Heiko Runz; Adolf Stiehl; Cyriel Y. Ponsioen; Cisca Wijmenga; Morten H. Vatn; Pieter C. F. Stokkers; Severine Vermeire; Christopher G. Mathew; Benedicte A. Lie; Ulrich Beuers; Michael P. Manns; Stefan Schreiber; Erik Schrumpf; Dieter Haeussinger; Andre Franke; Tom H. Karlsen

Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis

Johannes R. Hov, Verena Keitel, Jon K. Laerdahl, Lina Spomer, Eva Ellinghaus, Abdou ElSharawy, Espen Melum, Kirsten M. Boberg, Thomas Manke, Tobias Balschun, Christoph Schramm, Annika Bergquist, Tobias Weismueller, Daniel Gotthardt, Christian Rust, Liesbet Henckaerts, Clive M. Onnie, Rinse K. Weersma, Martina Sterneck, Andreas TeufelHeiko Runz, Adolf Stiehl, Cyriel Y. Ponsioen, Cisca Wijmenga, Morten H. Vatn, Pieter C. F. Stokkers, Severine Vermeire, Christopher G. Mathew, Benedicte A. Lie, Ulrich Beuers, Michael P. Manns, Stefan Schreiber, Erik Schrumpf, Dieter Haeussinger, Andre Franke, Tom H. Karlsen

Research output: Contribution to journal › Article › peer-review

118 Citations (Scopus)

Abstract

Background: TGR5, the G protein-coupled bile acid receptor 1 (GPBAR1), has been linked to inflammatory pathways as well as bile homeostasis, and could therefore be involved in primary sclerosing cholangitis (PSC) a chronic inflammatory bile duct disease. We aimed to extensively investigate TGR5 sequence variation in PSC, as well as functionally characterize detected variants. Methodology/Principal Findings: Complete resequencing of TGR5 was performed in 267 PSC patients and 274 healthy controls. Six nonsynonymous mutations were identified in addition to 16 other novel single-nucleotide polymorphisms. To investigate the impact from the nonsynonymous variants on TGR5, we created a receptor model, and introduced mutated TGR5 constructs into human epithelial cell lines. By using confocal microscopy, flow cytometry and a cAMP-sensitive luciferase assay, five of the nonsynonymous mutations (W83R, V178M, A217P, S272G and Q296X) were found to reduce or abolish TGR5 function. Fine-mapping of the previously reported PSC and UC associated locus at chromosome 2q35 in large patient panels revealed an overall association between the TGR5 single-nucleotide polymorphism rs11554825 and PSC (odds ratio = 1.14, 95% confidence interval: 1.03-1.26, p = 0.010) and UC (odds ratio = 1.19, 95% confidence interval 1.11-1.27, p = 8.5x10(-7)), but strong linkage disequilibrium precluded demarcation of TGR5 from neighboring genes. Conclusions/Significance: Resequencing of TGR5 along with functional investigations of novel variants provided unique insight into an important candidate gene for several inflammatory and metabolic conditions. While significant TGR5 associations were detected in both UC and PSC, further studies are needed to conclusively define the role of TGR5 variation in these diseases.

Original language	English
Article number	e12403
Journal	PL o S One
Volume	5
Issue number	8
Publication status	Published - 2010

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Hov, J. R., Keitel, V., Laerdahl, J. K., Spomer, L., Ellinghaus, E., ElSharawy, A., Melum, E., Boberg, K. M., Manke, T., Balschun, T., Schramm, C., Bergquist, A., Weismueller, T., Gotthardt, D., Rust, C., Henckaerts, L., Onnie, C. M., Weersma, R. K., Sterneck, M., ... Karlsen, T. H. (2010). Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis. PL o S One , 5(8), Article e12403.

@article{7766f6766ab04cc3b1c86e42b42e4cff,

title = "Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis",

abstract = "Background: TGR5, the G protein-coupled bile acid receptor 1 (GPBAR1), has been linked to inflammatory pathways as well as bile homeostasis, and could therefore be involved in primary sclerosing cholangitis (PSC) a chronic inflammatory bile duct disease. We aimed to extensively investigate TGR5 sequence variation in PSC, as well as functionally characterize detected variants. Methodology/Principal Findings: Complete resequencing of TGR5 was performed in 267 PSC patients and 274 healthy controls. Six nonsynonymous mutations were identified in addition to 16 other novel single-nucleotide polymorphisms. To investigate the impact from the nonsynonymous variants on TGR5, we created a receptor model, and introduced mutated TGR5 constructs into human epithelial cell lines. By using confocal microscopy, flow cytometry and a cAMP-sensitive luciferase assay, five of the nonsynonymous mutations (W83R, V178M, A217P, S272G and Q296X) were found to reduce or abolish TGR5 function. Fine-mapping of the previously reported PSC and UC associated locus at chromosome 2q35 in large patient panels revealed an overall association between the TGR5 single-nucleotide polymorphism rs11554825 and PSC (odds ratio = 1.14, 95% confidence interval: 1.03-1.26, p = 0.010) and UC (odds ratio = 1.19, 95% confidence interval 1.11-1.27, p = 8.5x10(-7)), but strong linkage disequilibrium precluded demarcation of TGR5 from neighboring genes. Conclusions/Significance: Resequencing of TGR5 along with functional investigations of novel variants provided unique insight into an important candidate gene for several inflammatory and metabolic conditions. While significant TGR5 associations were detected in both UC and PSC, further studies are needed to conclusively define the role of TGR5 variation in these diseases.",

author = "Hov, {Johannes R.} and Verena Keitel and Laerdahl, {Jon K.} and Lina Spomer and Eva Ellinghaus and Abdou ElSharawy and Espen Melum and Boberg, {Kirsten M.} and Thomas Manke and Tobias Balschun and Christoph Schramm and Annika Bergquist and Tobias Weismueller and Daniel Gotthardt and Christian Rust and Liesbet Henckaerts and Onnie, {Clive M.} and Weersma, {Rinse K.} and Martina Sterneck and Andreas Teufel and Heiko Runz and Adolf Stiehl and Ponsioen, {Cyriel Y.} and Cisca Wijmenga and Vatn, {Morten H.} and Stokkers, {Pieter C. F.} and Severine Vermeire and Mathew, {Christopher G.} and Lie, {Benedicte A.} and Ulrich Beuers and Manns, {Michael P.} and Stefan Schreiber and Erik Schrumpf and Dieter Haeussinger and Andre Franke and Karlsen, {Tom H.}",

year = "2010",

language = "English",

volume = "5",

journal = "PL o S One ",

issn = "1932-6203",

publisher = "Public Library of Science",

number = "8",

}

Hov, JR, Keitel, V, Laerdahl, JK, Spomer, L, Ellinghaus, E, ElSharawy, A, Melum, E, Boberg, KM, Manke, T, Balschun, T, Schramm, C, Bergquist, A, Weismueller, T, Gotthardt, D, Rust, C, Henckaerts, L, Onnie, CM, Weersma, RK, Sterneck, M, Teufel, A, Runz, H, Stiehl, A, Ponsioen, CY, Wijmenga, C, Vatn, MH, Stokkers, PCF, Vermeire, S, Mathew, CG, Lie, BA, Beuers, U, Manns, MP, Schreiber, S, Schrumpf, E, Haeussinger, D, Franke, A & Karlsen, TH 2010, 'Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis', PL o S One , vol. 5, no. 8, e12403.

TY - JOUR

T1 - Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis

AU - Hov, Johannes R.

AU - Keitel, Verena

AU - Laerdahl, Jon K.

AU - Spomer, Lina

AU - Ellinghaus, Eva

AU - ElSharawy, Abdou

AU - Melum, Espen

AU - Boberg, Kirsten M.

AU - Manke, Thomas

AU - Balschun, Tobias

AU - Schramm, Christoph

AU - Bergquist, Annika

AU - Weismueller, Tobias

AU - Gotthardt, Daniel

AU - Rust, Christian

AU - Henckaerts, Liesbet

AU - Onnie, Clive M.

AU - Weersma, Rinse K.

AU - Sterneck, Martina

AU - Teufel, Andreas

AU - Runz, Heiko

AU - Stiehl, Adolf

AU - Ponsioen, Cyriel Y.

AU - Wijmenga, Cisca

AU - Vatn, Morten H.

AU - Stokkers, Pieter C. F.

AU - Vermeire, Severine

AU - Mathew, Christopher G.

AU - Lie, Benedicte A.

AU - Beuers, Ulrich

AU - Manns, Michael P.

AU - Schreiber, Stefan

AU - Schrumpf, Erik

AU - Haeussinger, Dieter

AU - Franke, Andre

AU - Karlsen, Tom H.

PY - 2010

Y1 - 2010

N2 - Background: TGR5, the G protein-coupled bile acid receptor 1 (GPBAR1), has been linked to inflammatory pathways as well as bile homeostasis, and could therefore be involved in primary sclerosing cholangitis (PSC) a chronic inflammatory bile duct disease. We aimed to extensively investigate TGR5 sequence variation in PSC, as well as functionally characterize detected variants. Methodology/Principal Findings: Complete resequencing of TGR5 was performed in 267 PSC patients and 274 healthy controls. Six nonsynonymous mutations were identified in addition to 16 other novel single-nucleotide polymorphisms. To investigate the impact from the nonsynonymous variants on TGR5, we created a receptor model, and introduced mutated TGR5 constructs into human epithelial cell lines. By using confocal microscopy, flow cytometry and a cAMP-sensitive luciferase assay, five of the nonsynonymous mutations (W83R, V178M, A217P, S272G and Q296X) were found to reduce or abolish TGR5 function. Fine-mapping of the previously reported PSC and UC associated locus at chromosome 2q35 in large patient panels revealed an overall association between the TGR5 single-nucleotide polymorphism rs11554825 and PSC (odds ratio = 1.14, 95% confidence interval: 1.03-1.26, p = 0.010) and UC (odds ratio = 1.19, 95% confidence interval 1.11-1.27, p = 8.5x10(-7)), but strong linkage disequilibrium precluded demarcation of TGR5 from neighboring genes. Conclusions/Significance: Resequencing of TGR5 along with functional investigations of novel variants provided unique insight into an important candidate gene for several inflammatory and metabolic conditions. While significant TGR5 associations were detected in both UC and PSC, further studies are needed to conclusively define the role of TGR5 variation in these diseases.

AB - Background: TGR5, the G protein-coupled bile acid receptor 1 (GPBAR1), has been linked to inflammatory pathways as well as bile homeostasis, and could therefore be involved in primary sclerosing cholangitis (PSC) a chronic inflammatory bile duct disease. We aimed to extensively investigate TGR5 sequence variation in PSC, as well as functionally characterize detected variants. Methodology/Principal Findings: Complete resequencing of TGR5 was performed in 267 PSC patients and 274 healthy controls. Six nonsynonymous mutations were identified in addition to 16 other novel single-nucleotide polymorphisms. To investigate the impact from the nonsynonymous variants on TGR5, we created a receptor model, and introduced mutated TGR5 constructs into human epithelial cell lines. By using confocal microscopy, flow cytometry and a cAMP-sensitive luciferase assay, five of the nonsynonymous mutations (W83R, V178M, A217P, S272G and Q296X) were found to reduce or abolish TGR5 function. Fine-mapping of the previously reported PSC and UC associated locus at chromosome 2q35 in large patient panels revealed an overall association between the TGR5 single-nucleotide polymorphism rs11554825 and PSC (odds ratio = 1.14, 95% confidence interval: 1.03-1.26, p = 0.010) and UC (odds ratio = 1.19, 95% confidence interval 1.11-1.27, p = 8.5x10(-7)), but strong linkage disequilibrium precluded demarcation of TGR5 from neighboring genes. Conclusions/Significance: Resequencing of TGR5 along with functional investigations of novel variants provided unique insight into an important candidate gene for several inflammatory and metabolic conditions. While significant TGR5 associations were detected in both UC and PSC, further studies are needed to conclusively define the role of TGR5 variation in these diseases.

M3 - Article

SN - 1932-6203

VL - 5

JO - PL o S One

JF - PL o S One

IS - 8

M1 - e12403

ER -

Mutational Characterization of the Bile Acid Receptor TGR5 in Primary Sclerosing Cholangitis

Abstract

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