Circadian disruption synergizes with

alcohol in disrupting the gut/liver axis to promote liver disease

Christopher B Forsyth, PhD

Rush University Medical Center

Chicago, IL USA

Hypothesis

• Disruption of the intestinal barrier (i.e., Leaky Gut) is associated with numerous diseases, including metabolic syndrome, diabetes, cardiovascular disease, inflammatory bowel disease, Parkinson's disease and alcoholic liver disease

• We hypothesized that circadian disruption by light/dark shifting or genetics (ClockΔ19) would combine with alcohol to promote gut leakinessand alcoholic liver disease (ALD) in mice.

Gastroenterology, Volume 146, Issue 6, 2014, 1513 – 1524 (AMP=antimicrobial peptides)

Role for gut leakiness and intestinal microbiota in alcoholic liver disease (ALD).

Figure 1. Models of circadian disruption and chronic alcohol consumption experimental protocols.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 2. Genetic disruption of circadian organization increases intestinal permeability.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 3. Genetic disruption of circadian organization promotes alcohol-induced intestinal hyperpermeability.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 4. Serum lipopolysaccharide (LPS) levels are altered and affected by alcohol in ClockΔ19/Δ19 mutant mice.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 5. Cytoplasmic tight junction protein occludin levels in the proximal colon are significantly elevated in ClockΔ19/Δ19 mutant

mice.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 6. Genetic disruption of circadian organization promotes alcohol-induced hepatic steatosis.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 7. Environmental disruption of circadian organization increases intestinal permeability.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 8. Environmental disruption of circadian organization augments alcohol-induced intestinal hyperpermeability.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 9. Environmental disruption of circadian organization impacts serum LPS and LBP levels.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 10. Altered regulation of the tight junction protein occludin in the proximal colon by alcohol and chronic circadian disruption.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Figure 11. Environmental disruption of circadian organization promotes alcohol-induced liver pathology.

Summa KC, Voigt RM, Forsyth CB, Shaikh M, Cavanaugh K, et al. (2013) Disruption of the Circadian Clock in Mice Increases Intestinal Permeability and Promotes Alcohol-Induced Hepatic Pathology and Inflammation. PLoS ONE 8(6): e67102. doi:10.1371/journal.pone.0067102http://journals.plos.org/plosone/article?id=info:doi/10.1371/journal.pone.0067102

Conclusions

• Circadian disorganization, using independent genetic and environmental strategies, increases permeability of the intestinal epithelial barrier.

• Both genetic and environmental circadian disruption promote alcohol-induced gut leakiness, endotoxemia and steatohepatitis, possibly through a mechanism involving the tight junction protein occludin.

• Circadian disruption may therefore represent a previously unrecognized risk factor underlying the susceptibility to or development of alcoholic liver disease, as well as other conditions associated with intestinal hyperpermeability and an endotoxin-triggered inflammatory state.

Acknowledgements

• Ali Keshavarzian, MD Lab (Rush)• Robin Voigt-Zuwala, PhD• Christopher Forsyth, PhD• Maliha Shaikh, MS• Shiwen Song, MD• Yueming Tang, PhD• Phillip Engen, BS• Garth Swanson, MD• Faraz Bishehsari, MD• Lijuan Zhang, MD• Nailliw Preite, MS• Shohreh Raeisi, MS• Sherry Wilber

• Fred W. Turek, PhD Lab(Northwestern)• Keith Summa MD/PhD• Martha Vitaterna, PhD• Peng Jiang, PhD• Kate Cavanaugh

• Supported by NIH: NIAAA, NCRR and NCATS