Biology subjects

Potter, M.

Publications and source records attributed to Potter, M..

8 recordsLinked to original sources

bioRxiv2026

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↗

bioRxiv2026

AAV2 Crosslinks Actin Filaments: Implications for AAV Gene Therapy Vector Design

Adeno-associated virus (AAV) capsids are important gene therapy vectors, allowing for the one-time treatment of monogenetic disorders, with durable gene expression lasting for years. However, despite the clinical success of AAV usage, low transduction efficiencies require high dosage to achieve therapeutic efficacy, resulting in prohibitive costs and rare, but life-threatening immune responses. One key knowledge gap is how the capsids traffic to the nucleus following endocytosis. Here, we identify a direct interaction between AAV2 and actin filaments. Our results show that AAV2 bundles multiple actin filaments in a highly periodic hexagonal lattice. Therefore, we propose that AAV2 is the founding member of a new class of actin binding proteins, the actin bundling viruses (ABVs). Using cryogenic electron microscopy, we determined the structure of AAV2-actin filaments, identified the interaction interfaces, and engineered capsid variants that no longer bundle actin. Together, our results suggest that the AAV2-actin interaction may be responsible for trapping capsids and mediating peri-nuclear accumulation. Overall, this interaction expands the current understanding of AAV2 biology and offers new directions for capsid engineering.

biochemistry↗

bioRxiv2025

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↗

bioRxiv2024

Genetic variance in the murine defensin locus modulates glucose homeostasis

Insulin resistance is a heritable risk factor for many chronic diseases; however, the genetic drivers remain elusive. In seeking these, we performed genetic mapping of insulin sensitivity in 670 chow-fed Diversity Outbred in Australia (DOz) mice and identified a genome-wide significant locus (QTL) on chromosome 8 encompassing 17 defensin genes. By taking a systems genetics approach, we ultimately identified alpha-defensin 26 (Defa26) as the causal gene in this region. To validate these findings, we synthesized Defa26 and performed diet supplementation experiments in two mouse strains with distinct endogenous Defa26 expression levels. In the strain with relatively lower endogenous expression (C57BL/6J) supplementation improved insulin sensitivity and reduced gut permeability, while in the strain with higher endogenous expression (A/J) it caused hypoinsulinemia, glucose intolerance and muscle wasting. Based on gut microbiome and plasma bile acid profiling this appeared to be the result of disrupted microbial bile acid metabolism. These data illustrate the danger of single strain over-reliance and provide the first evidence of a link between host-genetics and insulin sensitivity which is mediated by the microbiome.

systems biology↗

bioRxiv2024

Identification of SEC61B as a novel regulator of calcium flux and platelet hyperreactivity in diabetes mellitus

High platelet reactivity is associated with adverse clinical events and is more frequent in people with diabetes mellitus (DM). To better understand platelet dysfunction in DM, we performed a proteomic analysis in platelets from a matched cohort of 34 people without, and 42 people with type 2 DM. The cohorts were matched by clinical characteristics including age, sex, and coronary artery disease burden. Using high sensitivity unbiased proteomics, we consistently identified over 2,400 intracellular proteins, and detected proteins that are differentially released by platelets from people with diabetes in response to low dose thrombin. Importantly, we identified the endoplasmic reticulum (ER) protein SEC61 translocon subunit beta (SEC61B) was increased in platelets from humans and mice with in vivo hyperglycemia. SEC61B was increased in megakaryocytes in mouse models of diabetes, in association with megakaryocyte ER stress. A rise in cytosolic calcium is a key aspect in platelet activation, and the SEC61 translocon is known to act as a channel for ER calcium leak. We demonstrate that cultured cells overexpressing SEC61B have increased calcium flux and decreased protein synthesis. In accordance, hyperglycemic mouse platelets mobilized more calcium to the cytosol and had lower protein synthesis compared with normoglycemic platelets. Independently, in vitro induction of ER stress increased platelet SEC61B expression and markers of platelet activation. We propose a mechanism whereby ER stress-induced upregulation of platelet SEC61B leads to increased cytosolic calcium, potentially contributing to platelet hyperactivity in people with diabetes. Key PointsO_LIPlatelet SEC61B is increased in hyperglycemia and contributes to increased endoplasmic reticulum (ER) calcium leak C_LIO_LIIncreased ER calcium leak is associated with ER stress and platelet hyperactivity C_LI

cell biology↗

bioRxiv2023

Assessing the validity of the zero-velocity update method for sprinting speeds

The zero-velocity update (ZUPT) method has become a popular approach to estimate foot kinematics from foot worn inertial measurement units (IMUs) during walking and running. However, the accuracy of the ZUPT method for stride parameters at sprinting speeds remains unknown, specifically when using sensors with characteristics well suited for sprinting (i.e., high accelerometer and gyroscope ranges and sampling rates). Seventeen participants performed 70-meter track sprints while wearing a Blue Trident IMeasureU IMU. Two cameras, at 20 and 70 meters from the start, were used to validate the ZUPT method on a stride-by-stride and on a cumulative distance basis. In particular, the validity of the ZUPT method was assessed for: (1) estimating a single stride length attained at the end of a 70m sprint (i.e., stride at 70m); (2) estimating cumulative distance from [~]20 to [~]70 m; and (3) estimating total distance traveled for a 70-meter track sprint. Individual stride length errors at the 70-meter mark were within - 6% to 3%, with a bias of -0.27%. Cumulative distance errors were within -4 to 2%, with biases ranging from -0.85 to -1.22%. The results of this study demonstrate the ZUPT method provides accurate estimates of stride length and cumulative distance traveled for sprinting speeds.

bioengineering↗

bioRxiv2023

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↗

bioRxiv2023

Leveraging genetic diversity to identify small molecules that reverse mouse skeletal muscle insulin resistance

Systems genetics has begun to tackle the complexity of insulin resistance by capitalising on computational advances to study high-diversity populations. "Diversity Outbred in Australia (DOz)" is a population of genetically unique mice with profound metabolic heterogeneity. We leveraged this variance to explore skeletal muscles contribution to whole-body insulin action through metabolic phenotyping and skeletal muscle proteomics of 215 DOz mice. Linear modelling identified 553 proteins that associated with whole-body insulin sensitivity (Matsuda Index) including regulators of endocytosis and muscle proteostasis. To enrich for causality, we refined this network by focussing on negatively associated, genetically regulated proteins, resulting in a 76-protein fingerprint of insulin resistance. We sought to perturb this network and restore insulin action with small molecules by integrating the Broad Institute Connectivity Map platform and in vitro assays of insulin action using the Prestwick chemical library. These complimentary approaches identified the antibiotic thiostrepton as an insulin resistance reversal agent. Subsequent validation in ex vivo insulin resistant mouse muscle, and palmitate induced insulin resistant myotubes demonstrated potent insulin action restoration, potentially via up-regulation of glycolysis. This work demonstrates the value of a drug-centric framework to validate systems level analysis by identifying potential therapeutics for insulin resistance.

systems biology↗