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Identification of a genetic locus controlling bacteria-driven colitis and associated cancer through effects on innate inflammation.

Boulard O, Kirchberger S, Royston DJ, Maloy KJ, Powrie FM - J. Exp. Med. (2012)

Bottom Line: Chronic inflammation of the intestine has been associated with an elevated risk of developing colorectal cancer.Using a tumor-promoting model combining chronic Helicobacter hepaticus infection and the carcinogen azoxymethane, we found that Hiccs also regulates the frequency of colitis-associated neoplasia.Our study highlights the importance of innate immune cells and their genetic configuration in driving progression from inflammation toward cancer and opens the door for analysis of these pathways in human inflammatory disorders and associated cancers.

View Article: PubMed Central - HTML - PubMed

Affiliation: Translational Gastroenterology Unit, Nuffield Department of Clinical Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, England, UK.

ABSTRACT
Chronic inflammation of the intestine has been associated with an elevated risk of developing colorectal cancer. Recent association studies have highlighted the role of genetic predisposition in the etiology of colitis and started to unravel its complexity. However, the genetic factors influencing the progression from colon inflammation to tumorigenesis are not known. We report the identification of a genetic interval Hiccs that regulates Helicobacter hepaticus-induced colitis and associated cancer susceptibility in a 129.RAG(-/-) mouse model. The 1.7-Mb congenic interval on chromosome 3, containing eight genes and five microRNAs, renders susceptible mice resistant to colitis and reduces tumor incidence and multiplicity. Bone marrow chimera experiments showed that resistance is conferred by the hematopoietic compartment. Moreover, the Hiccs locus controls the induction of the innate inflammatory response by regulating cytokine expression and granulocyte recruitment by Thy1(+) innate lymphoid cells. Using a tumor-promoting model combining chronic Helicobacter hepaticus infection and the carcinogen azoxymethane, we found that Hiccs also regulates the frequency of colitis-associated neoplasia. Our study highlights the importance of innate immune cells and their genetic configuration in driving progression from inflammation toward cancer and opens the door for analysis of these pathways in human inflammatory disorders and associated cancers.

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The Hiccs locus controls the early inflammatory response through ILCs. (A–D) Time course of early H. hepaticus infection in susceptible 129.RAG (gray circles) and protected R17 (open diamonds) mice. (A) Spleen weight and total colonic LPL count. (B) Frequencies and absolute numbers of granulocytes (CD11bhi Gr1hi), DC (CD11chi CD11b+/−), and monocytes/macrophages (CD11bhi Gr1−/int CD11c−) in colon LPL. For ILC (lin neg, Thy1hi, Sca1hi), absolute numbers are shown. For each time point and genotype, the mean and SEM of five to seven mice pooled from two independent experiments are shown. (C) Representative FACS plot of 129.RAG and R17 colonic LPL, showing granulocyte gate (CD11bhi Gr1hi) and frequency. (D) TNF, IL-1β, IFN-γ, IL-17A, and IL-22 were detected by FlowCytomix in the supernatants of overnight cultured LPL. Each dot represents the mean and SEM of two pools with two mice each. IL-23p19 mRNA expression was analyzed by Q-PCR of RNA isolated from fresh LPL. Each dot represents the mean and SEM of two to four mice. One representative experiment out of two with similar results is shown. (E and F) 129.RAG and R17 mice were treated with isotype control or anti-Thy1 antibody to deplete ILC at days 6 and 0 before H. hepaticus infection. Mice were analyzed at day 6 after infection. One of two independent experiments with similar results is shown. (E) Total LPL number, granulocyte frequency, and absolute number. Horizontal lines represent means. (F) TNF, IL-1β, and IFN-γ were detected by FlowCytomix in the supernatants of overnight cultured LPL. Mean and SEM are shown for three mice per condition. **, P < 0.01; *, P < 0.05, Mann-Whitney nonparametric test for A, B, and E, unpaired Student’s t test for F.
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fig7: The Hiccs locus controls the early inflammatory response through ILCs. (A–D) Time course of early H. hepaticus infection in susceptible 129.RAG (gray circles) and protected R17 (open diamonds) mice. (A) Spleen weight and total colonic LPL count. (B) Frequencies and absolute numbers of granulocytes (CD11bhi Gr1hi), DC (CD11chi CD11b+/−), and monocytes/macrophages (CD11bhi Gr1−/int CD11c−) in colon LPL. For ILC (lin neg, Thy1hi, Sca1hi), absolute numbers are shown. For each time point and genotype, the mean and SEM of five to seven mice pooled from two independent experiments are shown. (C) Representative FACS plot of 129.RAG and R17 colonic LPL, showing granulocyte gate (CD11bhi Gr1hi) and frequency. (D) TNF, IL-1β, IFN-γ, IL-17A, and IL-22 were detected by FlowCytomix in the supernatants of overnight cultured LPL. Each dot represents the mean and SEM of two pools with two mice each. IL-23p19 mRNA expression was analyzed by Q-PCR of RNA isolated from fresh LPL. Each dot represents the mean and SEM of two to four mice. One representative experiment out of two with similar results is shown. (E and F) 129.RAG and R17 mice were treated with isotype control or anti-Thy1 antibody to deplete ILC at days 6 and 0 before H. hepaticus infection. Mice were analyzed at day 6 after infection. One of two independent experiments with similar results is shown. (E) Total LPL number, granulocyte frequency, and absolute number. Horizontal lines represent means. (F) TNF, IL-1β, and IFN-γ were detected by FlowCytomix in the supernatants of overnight cultured LPL. Mean and SEM are shown for three mice per condition. **, P < 0.01; *, P < 0.05, Mann-Whitney nonparametric test for A, B, and E, unpaired Student’s t test for F.

Mentions: To gain further insight into how Hiccs controls the early inflammatory response, we performed a kinetic analysis of the innate response after H. hepaticus infection in protected and susceptible mice. As early as 3 to 8 d after H. hepaticus infection of susceptible 129.RAG mice, we observed a small increase in number of colonic LPL, as well as signs of systemic inflammation, identified by an increased spleen weight (Fig. 7 A). By day 14 after infection, clear differences in spleen weight and LPL numbers were apparent between susceptible 129.RAG and protected R17 mice (Fig. 7 A).


Identification of a genetic locus controlling bacteria-driven colitis and associated cancer through effects on innate inflammation.

Boulard O, Kirchberger S, Royston DJ, Maloy KJ, Powrie FM - J. Exp. Med. (2012)

The Hiccs locus controls the early inflammatory response through ILCs. (A–D) Time course of early H. hepaticus infection in susceptible 129.RAG (gray circles) and protected R17 (open diamonds) mice. (A) Spleen weight and total colonic LPL count. (B) Frequencies and absolute numbers of granulocytes (CD11bhi Gr1hi), DC (CD11chi CD11b+/−), and monocytes/macrophages (CD11bhi Gr1−/int CD11c−) in colon LPL. For ILC (lin neg, Thy1hi, Sca1hi), absolute numbers are shown. For each time point and genotype, the mean and SEM of five to seven mice pooled from two independent experiments are shown. (C) Representative FACS plot of 129.RAG and R17 colonic LPL, showing granulocyte gate (CD11bhi Gr1hi) and frequency. (D) TNF, IL-1β, IFN-γ, IL-17A, and IL-22 were detected by FlowCytomix in the supernatants of overnight cultured LPL. Each dot represents the mean and SEM of two pools with two mice each. IL-23p19 mRNA expression was analyzed by Q-PCR of RNA isolated from fresh LPL. Each dot represents the mean and SEM of two to four mice. One representative experiment out of two with similar results is shown. (E and F) 129.RAG and R17 mice were treated with isotype control or anti-Thy1 antibody to deplete ILC at days 6 and 0 before H. hepaticus infection. Mice were analyzed at day 6 after infection. One of two independent experiments with similar results is shown. (E) Total LPL number, granulocyte frequency, and absolute number. Horizontal lines represent means. (F) TNF, IL-1β, and IFN-γ were detected by FlowCytomix in the supernatants of overnight cultured LPL. Mean and SEM are shown for three mice per condition. **, P < 0.01; *, P < 0.05, Mann-Whitney nonparametric test for A, B, and E, unpaired Student’s t test for F.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3405508&req=5

fig7: The Hiccs locus controls the early inflammatory response through ILCs. (A–D) Time course of early H. hepaticus infection in susceptible 129.RAG (gray circles) and protected R17 (open diamonds) mice. (A) Spleen weight and total colonic LPL count. (B) Frequencies and absolute numbers of granulocytes (CD11bhi Gr1hi), DC (CD11chi CD11b+/−), and monocytes/macrophages (CD11bhi Gr1−/int CD11c−) in colon LPL. For ILC (lin neg, Thy1hi, Sca1hi), absolute numbers are shown. For each time point and genotype, the mean and SEM of five to seven mice pooled from two independent experiments are shown. (C) Representative FACS plot of 129.RAG and R17 colonic LPL, showing granulocyte gate (CD11bhi Gr1hi) and frequency. (D) TNF, IL-1β, IFN-γ, IL-17A, and IL-22 were detected by FlowCytomix in the supernatants of overnight cultured LPL. Each dot represents the mean and SEM of two pools with two mice each. IL-23p19 mRNA expression was analyzed by Q-PCR of RNA isolated from fresh LPL. Each dot represents the mean and SEM of two to four mice. One representative experiment out of two with similar results is shown. (E and F) 129.RAG and R17 mice were treated with isotype control or anti-Thy1 antibody to deplete ILC at days 6 and 0 before H. hepaticus infection. Mice were analyzed at day 6 after infection. One of two independent experiments with similar results is shown. (E) Total LPL number, granulocyte frequency, and absolute number. Horizontal lines represent means. (F) TNF, IL-1β, and IFN-γ were detected by FlowCytomix in the supernatants of overnight cultured LPL. Mean and SEM are shown for three mice per condition. **, P < 0.01; *, P < 0.05, Mann-Whitney nonparametric test for A, B, and E, unpaired Student’s t test for F.
Mentions: To gain further insight into how Hiccs controls the early inflammatory response, we performed a kinetic analysis of the innate response after H. hepaticus infection in protected and susceptible mice. As early as 3 to 8 d after H. hepaticus infection of susceptible 129.RAG mice, we observed a small increase in number of colonic LPL, as well as signs of systemic inflammation, identified by an increased spleen weight (Fig. 7 A). By day 14 after infection, clear differences in spleen weight and LPL numbers were apparent between susceptible 129.RAG and protected R17 mice (Fig. 7 A).

Bottom Line: Chronic inflammation of the intestine has been associated with an elevated risk of developing colorectal cancer.Using a tumor-promoting model combining chronic Helicobacter hepaticus infection and the carcinogen azoxymethane, we found that Hiccs also regulates the frequency of colitis-associated neoplasia.Our study highlights the importance of innate immune cells and their genetic configuration in driving progression from inflammation toward cancer and opens the door for analysis of these pathways in human inflammatory disorders and associated cancers.

View Article: PubMed Central - HTML - PubMed

Affiliation: Translational Gastroenterology Unit, Nuffield Department of Clinical Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, England, UK.

ABSTRACT
Chronic inflammation of the intestine has been associated with an elevated risk of developing colorectal cancer. Recent association studies have highlighted the role of genetic predisposition in the etiology of colitis and started to unravel its complexity. However, the genetic factors influencing the progression from colon inflammation to tumorigenesis are not known. We report the identification of a genetic interval Hiccs that regulates Helicobacter hepaticus-induced colitis and associated cancer susceptibility in a 129.RAG(-/-) mouse model. The 1.7-Mb congenic interval on chromosome 3, containing eight genes and five microRNAs, renders susceptible mice resistant to colitis and reduces tumor incidence and multiplicity. Bone marrow chimera experiments showed that resistance is conferred by the hematopoietic compartment. Moreover, the Hiccs locus controls the induction of the innate inflammatory response by regulating cytokine expression and granulocyte recruitment by Thy1(+) innate lymphoid cells. Using a tumor-promoting model combining chronic Helicobacter hepaticus infection and the carcinogen azoxymethane, we found that Hiccs also regulates the frequency of colitis-associated neoplasia. Our study highlights the importance of innate immune cells and their genetic configuration in driving progression from inflammation toward cancer and opens the door for analysis of these pathways in human inflammatory disorders and associated cancers.

Show MeSH
Related in: MedlinePlus