Cystathionine γ-lyase exacerbates Helicobacter pylori immunopathogenesis by promoting macrophage metabolic remodeling and activation
Yvonne L. Latour, Johanna C. Sierra, Jordan L. Finley, Mohammad Asim, Daniel P. Barry, Margaret M. Allaman, Thaddeus M. Smith, Kara M. McNamara, Paula B. Luis, Claus Schneider, Justin Jacobse, Jeremy A. Goettel, M. Wade Calcutt, Kristie L. Rose, Kevin L. Schey, Ginger L. Milne, Alberto G. Delgado, M. Blanca Piazuelo, Bindu D. Paul, Solomon H. Snyder, Alain P. Gobert, Keith T. Wilson
Yvonne L. Latour, Johanna C. Sierra, Jordan L. Finley, Mohammad Asim, Daniel P. Barry, Margaret M. Allaman, Thaddeus M. Smith, Kara M. McNamara, Paula B. Luis, Claus Schneider, Justin Jacobse, Jeremy A. Goettel, M. Wade Calcutt, Kristie L. Rose, Kevin L. Schey, Ginger L. Milne, Alberto G. Delgado, M. Blanca Piazuelo, Bindu D. Paul, Solomon H. Snyder, Alain P. Gobert, Keith T. Wilson
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Research Article Gastroenterology Immunology

Cystathionine γ-lyase exacerbates Helicobacter pylori immunopathogenesis by promoting macrophage metabolic remodeling and activation

Abstract

Macrophages play a crucial role in the inflammatory response to the human stomach pathogen Helicobacter pylori, which infects half of the world’s population and causes gastric cancer. Recent studies have highlighted the importance of macrophage immunometabolism in their activation state and function. We have demonstrated that the cysteine-producing enzyme cystathionine γ-lyase (CTH) is upregulated in humans and mice with H. pylori infection. Here, we show that induction of CTH in macrophages by H. pylori promoted persistent inflammation. Cth–/– mice had reduced macrophage and T cell activation in H. pylori–infected tissues, an altered metabolome, and decreased enrichment of immune-associated gene networks, culminating in decreased H. pylori–induced gastritis. CTH is downstream of the proposed antiinflammatory molecule, S-adenosylmethionine (SAM). Whereas Cth–/– mice exhibited gastric SAM accumulation, WT mice treated with SAM did not display protection against H. pylori–induced inflammation. Instead, we demonstrated that Cth-deficient macrophages exhibited alterations in the proteome, decreased NF-κB activation, diminished expression of macrophage activation markers, and impaired oxidative phosphorylation and glycolysis. Thus, through altering cellular respiration, CTH is a key enhancer of macrophage activation, contributing to a pathogenic inflammatory response that is the universal precursor for the development of H. pylori–induced gastric disease.

Authors

Yvonne L. Latour, Johanna C. Sierra, Jordan L. Finley, Mohammad Asim, Daniel P. Barry, Margaret M. Allaman, Thaddeus M. Smith, Kara M. McNamara, Paula B. Luis, Claus Schneider, Justin Jacobse, Jeremy A. Goettel, M. Wade Calcutt, Kristie L. Rose, Kevin L. Schey, Ginger L. Milne, Alberto G. Delgado, M. Blanca Piazuelo, Bindu D. Paul, Solomon H. Snyder, Alain P. Gobert, Keith T. Wilson

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Figure 7

CTH suppresses DNA methylation and supports M1 macrophage gene expression.

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CTH suppresses DNA methylation and supports M1 macrophage gene expressio...
(A) Expression of Cth by WT BMmacs 24 hours after challenge; n = 4–10 biological replicates from 3 independent experiments. (B) Representative plot of CD11b and F4/80 expression of WT and Cth–/– BMmacs 24 hours p.i. with H. pylori. (C) Fold change of metabolite levels in BMmacs 24 hours p.i. with H. pylori compared with control (ctrl); n = 4 biological replicates. (D) Representative plot and (E) MFI quantification of 5mC staining in WT and Cth–/– BMmacs 6 hours p.i. with H. pylori; n = 4 biological replicates. (F) Expression of proinflammatory macrophage activation markers by WT and Cth–/– BMmacs 6 hours p.i. with H. pylori; n = 4–10 biological replicates per genotype from 3 independent experiments. (G) NOS2 protein immunoblot and densitometric analysis of WT and Cth–/– BMmacs 24 hours p.i. with H. pylori; n = 4 biological replicates per genotype. (H) NO2– concentration in supernatants from WT and Cth–/– BMmacs; n = 8 biological replicates from 2 independent experiments. (I) Expression of antiinflammatory macrophage activation markers by WT and Cth–/– BMmacs 6 hours p.i. with H. pylori; n = 4–10 biological replicates per genotype from 3 independent experiments. (J) Phosphorylated RELA (pRELA) protein immunoblot and densitometric analysis of WT and Cth–/– BMmacs 30 minutes p.i. with H. pylori; n = 3–4 biological replicates from 2 independent experiments. All values are reported as mean ± SEM. Statistical analyses, where shown: (A) 1-way ANOVA with Dunnett’s test; (C) 1-way ANOVA with Newman-Keuls post hoc test; (E, F, H, and I) 1-way ANOVA with Kruskal-Wallis test, followed by a Mann-Whitney U test; (J) Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001 vs. uninfected; §P < 0.05, §§P < 0.01 vs. WT.