Translational characterization of the temporal dynamics of metabolic dysfunctions in liver, adipose tissue and the gut during diet-induced NASH development in Ldlr-/-.Leiden mice

Eveline Gart; Wim van Duyvenvoorde; Jessica M Snabel; Christa de Ruiter; Joline Attema; Martien P M Caspers; Serene Lek; Bertie Joan van Heuven; Arjen G C L Speksnijder; Martin Giera; Aswin Menke; Kanita Salic; Kendra K Bence; Gregory J Tesz; Jaap Keijer; Robert Kleemann; Martine C Morrison

doi:10.1016/j.heliyon.2023.e13985

Translational characterization of the temporal dynamics of metabolic dysfunctions in liver, adipose tissue and the gut during diet-induced NASH development in Ldlr-/-.Leiden mice

Heliyon. 2023 Feb 24;9(3):e13985. doi: 10.1016/j.heliyon.2023.e13985. eCollection 2023 Mar.

Authors

Eveline Gart^{1

2}, Wim van Duyvenvoorde¹, Jessica M Snabel¹, Christa de Ruiter¹, Joline Attema¹, Martien P M Caspers³, Serene Lek⁴, Bertie Joan van Heuven⁵, Arjen G C L Speksnijder⁵, Martin Giera⁶, Aswin Menke¹, Kanita Salic¹, Kendra K Bence⁷, Gregory J Tesz⁷, Jaap Keijer², Robert Kleemann¹, Martine C Morrison¹

Affiliations

¹ Department of Metabolic Health Research, The Netherlands Organization for Applied Scientific Research (TNO), 2333 CK Leiden, the Netherlands.
² Human and Animal Physiology, Wageningen University, 6708 WD Wageningen, the Netherlands.
³ Department of Microbiology and Systems Biology, The Netherlands Organization for Applied Scientific Research (TNO), Zeist, the Netherlands.
⁴ Clinnovate Health UK Ltd, Glasgow, United Kingdom.
⁵ Naturalis Biodiversity Center, Leiden, the Netherlands.
⁶ Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands.
⁷ Pfizer Worldwide Research, Development & Medical, Internal Medicine Research Unit, Cambridge, MA, USA.

Abstract

Background: NAFLD progression, from steatosis to inflammation and fibrosis, results from an interplay of intra- and extrahepatic mechanisms. Disease drivers likely include signals from white adipose tissue (WAT) and gut. However, the temporal dynamics of disease development remain poorly understood.

Methods: High-fat-diet (HFD)-fed Ldlr-/-.Leiden mice were compared to chow-fed controls. At t = 0, 8, 16, 28 and 38w mice were euthanized, and liver, WAT depots and gut were analyzed biochemically, histologically and by lipidomics and transcriptomics together with circulating factors to investigate the sequence of pathogenic events and organ cross-talk during NAFLD development.

Results: HFD-induced obesity was associated with an increase in visceral fat, plasma lipids and hyperinsulinemia at t = 8w, along with increased liver steatosis and circulating liver damage biomarkers. In parallel, upstream regulator analysis predicted that lipid catabolism regulators were deactivated and lipid synthesis regulators were activated. Subsequently, hepatocyte hypertrophy, oxidative stress and hepatic inflammation developed. Hepatic collagen accumulated from t = 16 w and became pronounced at t = 28-38 w. Epididymal WAT was maximally hypertrophic from t = 8 w, which coincided with inflammation development. Mesenteric and subcutaneous WAT hypertrophy developed slower and did not appear to reach a maximum, with minimal inflammation. In gut, HFD significantly increased permeability, induced a shift in microbiota composition from t = 8 w and changed circulating gut-derived metabolites.

Conclusion: HFD-fed Ldlr-/-.Leiden mice develop obesity, dyslipidemia and insulin resistance, essentially as observed in obese NAFLD patients, underlining their translational value. We demonstrate that marked epididymal-WAT inflammation, and gut permeability and dysbiosis precede the development of NAFLD stressing the importance of a multiple-organ approach in the prevention and treatment of NAFLD.

Keywords: Inter-organ crosstalk; Liver fibrosis; Non-alcoholic fatty liver disease; Oxidative stress; Temporal dynamics.