bioRxiv Science⌕ Search

Biology subjects

Masson, S. W. C.

Publications and source records attributed to Masson, S. W. C..

3 recordsLinked to original sources

Systems genetics identifies ETS1 as a stress-dependent regulator of adipocyte insulin action and heme-iron homeostasis

White adipose tissue plays a central role in systemic energy homeostasis by buffering nutrient excess through insulin-stimulated glucose uptake and triglyceride storage. Despite its importance, the genetic and molecular mechanisms governing adipose tissue insulin action remain poorly defined because tissue-specific insulin responsiveness has been difficult to quantify at the scale required for genetic discovery. Here, we developed the first scalable platform for high-throughput genetic mapping of tissue-specific insulin action in adipose tissue, enabling systems genetic analysis across 559 genetically diverse Diversity Outbred Australia (DOz) mice. Genetic analysis accounting for adiposity identified 39 loci associated with adipose tissue insulin action, demonstrating that adipose insulin responsiveness is a genetically encoded trait that captures a dimension of metabolic health beyond adiposity. Among these, a strong diet-dependent locus on chromosome 9 encompassed the transcription factor Ets1. Functional studies demonstrated that Ets1 silencing selectively restored insulin-stimulated glucose uptake in insulin-resistant adipocytes. Proteomic profiling revealed that ETS1 orchestrates a stress-responsive program involving heme metabolism, iron handling and redox homeostasis. Consistent with this, ETS1 knockdown reduced cellular heme and labile iron levels and attenuated oxidative stress under insulin-resistant conditions. Collectively, these findings demonstrate the power of systems genetics to identify previously unrecognised regulators of adipose insulin action and establish the heme-iron axis as a critical determinant of adipocyte insulin responsiveness.

systems biology↗

Very long-chain ceramides in muscle associate with insulin resistance independent of obesity

Lipids, in particular ceramides and diacylglycerols (DAGs), are implicated in insulin resistance, however their precise roles remain unclear. We leverage natural genetic variation to examine muscle lipids and systemic insulin resistance (IR) in 399 Diversity Outbred Australia mice. Adipose mass was associated with 55% of muscle lipids and IR, with DAGs as the only enriched lipid class. To disentangle adiposity and muscle lipid contributions, we used two approaches: (1) linear modelling of muscle lipids corrected for adipose mass on systemic IR, and (2) stratifying mice into insulin sensitivity quartiles within adiposity bins. Both revealed that very long-chain ceramides, but not DAGs, were linked to IR. Transcriptomic and proteomics further associated these ceramides with cellular and mitochondrial stress. DAGs correlated with leptin expression in muscle, suggesting they originate from muscle-residing adipocytes. We propose that many muscle lipids, including DAGs, associate with IR due to adipose accumulation rather than directly influencing muscle insulin sensitivity. By addressing the relationship between adiposity and metabolic state, we identified very long-chain muscle ceramides as highly associated with IR independently of adiposity.

systems biology↗

The genetic and dietary landscape of the muscle insulin signalling network

Metabolic disease is caused by a combination of genetic and environmental factors, yet few studies have examined how these factors influence signal transduction, a key mediator of metabolism. Using mass spectrometry-based phosphoproteomics, we quantified 23,126 phosphosites in skeletal muscle of five genetically distinct mouse strains in two dietary environments, with and without acute in vivo insulin stimulation. Almost half of the insulin-regulated phosphoproteome was modified by genetic background on an ordinary diet, and high-fat high-sugar feeding affected insulin signalling in a strain-dependent manner. Our data revealed coregulated subnetworks within the insulin signalling pathway, expanding our understanding of the pathways organisation. Furthermore, associating diverse signalling responses with insulin-stimulated glucose uptake uncovered regulators of muscle insulin responsiveness, including the regulatory phosphosite S469 on Pfkfb2, a key activator of glycolysis. Finally, we confirmed the role of glycolysis in modulating insulin action in insulin resistance. Our results underscore the significance of genetics in shaping global signalling responses and their adaptability to environmental changes, emphasizing the utility of studying biological diversity with phosphoproteomics to discover key regulatory mechanisms of complex traits.

systems biology↗