Melbourne Medical School Collected Works - Research Publications

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    Interactive calculator to estimate insulin sensitivity in type 1 diabetes
    Januszewski, AS ; Niedzwiecki, P ; Sachithanandan, N ; Ward, GM ; O'Neal, DN ; Zozulinska-Ziolkiewicz, DA ; Uruska, AA ; Jenkins, AJ (WILEY, 2024-05)
    The gold standard for measuring insulin sensitivity (IS) is the hyperinsulinemic-euglycemic clamp, a time, costly, and labor-intensive research tool. A low insulin sensitivity is associated with a complication-risk in type 1 diabetes. Various formulae using clinical data have been developed and correlated with measured IS in type 1 diabetes. We consolidated multiple formulae into an online calculator (bit.ly/estimated-GDR), enabling comparison of IS and its probability of IS <4.45 mg/kg/min (low) or >6.50 mg/kg/min (high), as measured in a validation set of clamps in 104 adults with type 1 diabetes. Insulin sensitivity calculations using different formulae varied significantly, with correlations (R2) ranging 0.005-0.87 with agreement in detecting low and high glucose disposal rates in the range 49-93% and 89-100%, respectively. We demonstrate that although the calculated IS varies between formulae, their interpretation remains consistent. Our free online calculator offers a user-friendly tool for individual IS calculations and also offers efficient batch processing of data for research.
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    A 3D printed flow sensor for microfluidic applications
    Hawke, A ; Concilia, G ; Thurgood, P ; Ahnood, A ; Baratchi, S ; Khoshmanesh, K (ELSEVIER SCIENCE SA, 2023-11-01)
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    Endothelial Response to the Combined Biomechanics of Vessel Stiffness and Shear Stress Is Regulated via Piezo1
    Lai, A ; Zhou, Y ; Thurgood, P ; Chheang, C ; Sekar, NC ; Nguyen, N ; Peter, K ; Khoshmanesh, K ; Baratchi, S (AMER CHEMICAL SOC, 2023-12-11)
    How endothelial cells sense and respond to dynamic changes in their biophysical surroundings as we age is not fully understood. Vascular stiffness is clearly a contributing factor not only in several cardiovascular diseases but also in physiological processes such as aging and vascular dementia. To address this gap, we utilized a microfluidic model to explore how substrate stiffness in the presence of shear stress affects endothelial morphology, senescence, proliferation, and inflammation. We also studied the role of mechanosensitive ion channel Piezo1 in endothelial responses under the combined effect of shear stress and substrate stiffness. To do so, we cultured endothelial cells inside microfluidic channels covered with fibronectin-coated elastomer with elastic moduli of 40 and 200 kPa, respectively, mimicking the stiffness of the vessel walls in young and aged arteries. The endothelial cells were exposed to atheroprotective and atherogenic shear stress levels of 10 and 2 dyn/cm2, respectively. Our findings show that substrate stiffness affects senescence under atheroprotective flow conditions and cytoskeleton remodeling, senescence, and inflammation under atherogenic flow conditions. Additionally, we found that the expression of Piezo1 plays a crucial role in endothelial adaptation to flow and regulation of inflammation under both atheroprotective and atherogenic shear stress levels. However, Piezo1 contribution to endothelial senescence was limited to the soft substrate and atheroprotective shear stress level. Overall, our study characterizes the response of endothelial cells to the combined effect of shear stress and substrate stiffness and reveals a previously unidentified role of Piezo1 in endothelial response to vessel stiffening, which potentially can be therapeutically targeted to alleviate endothelial dysfunction in aging adults.
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    A microfluidic model to study the effects of arrhythmic flows on endothelial cells
    Lai, A ; Hawke, A ; Mohammed, M ; Thurgood, P ; Concilia, G ; Peter, K ; Khoshmanesh, K ; Baratchi, S (Royal Society of Chemistry, 2024-03-21)
    Atrial fibrillation (AF) is the most common type of cardiac arrhythmia and an important contributor to morbidity and mortality. Endothelial dysfunction has been postulated to be an important contributing factor in cardiovascular events in patients with AF. However, how vascular endothelial cells respond to arrhythmic flow is not fully understood, mainly due to the limitation of current in vitro systems to mimic arrhythmic flow conditions. To address this limitation, we developed a microfluidic system to study the effect of arrhythmic flow on the mechanobiology of human aortic endothelial cells (HAECs). The system utilises a computer-controlled piezoelectric pump for generating arrhythmic flow with a unique ability to control the variability in both the frequency and amplitude of pulse waves. The flow rate is modulated to reflect physiological or pathophysiological shear stress levels on endothelial cells. This enabled us to systematically dissect the importance of variability in the frequency and amplitude of pulses and shear stress level on endothelial cell mechanobiology. Our results indicated that arrhythmic flow at physiological shear stress level promotes endothelial cell spreading and reduces the plasma membrane-to-cytoplasmic distribution of β-catenin. In contrast, arrhythmic flow at low and atherogenic shear stress levels does not promote endothelial cell spreading or redistribution of β-catenin. Interestingly, under both shear stress levels, arrhythmic flow induces inflammation by promoting monocyte adhesion via an increase in ICAM-1 expression. Collectively, our microfluidic system provides opportunities to study the effect of arrhythmic flows on vascular endothelial mechanobiology in a systematic and reproducible manner.
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    Epigenetic Changes in Diabetes and Cardiovascular Risk
    Keating, ST ; Plutzky, J ; El-Osta, A (LIPPINCOTT WILLIAMS & WILKINS, 2016-05-27)
    Cardiovascular complications remain the leading causes of morbidity and premature mortality in patients with diabetes mellitus. Studies in humans and preclinical models demonstrate lasting gene expression changes in the vasculopathies initiated by previous exposure to high glucose concentrations and the associated overproduction of reactive oxygen species. The molecular signatures of chromatin architectures that sensitize the genome to these and other cardiometabolic risk factors of the diabetic milieu are increasingly implicated in the biological memory underlying cardiovascular complications and now widely considered as promising therapeutic targets. Atherosclerosis is a complex heterocellular disease where the contributing cell types possess distinct epigenomes shaping diverse gene expression. Although the extent that pathological chromatin changes can be manipulated in human cardiovascular disease remains to be established, the clinical applicability of epigenetic interventions will be greatly advanced by a deeper understanding of the cell type-specific roles played by writers, erasers, and readers of chromatin modifications in the diabetic vasculature. This review details a current perspective of epigenetic mechanisms of macrovascular disease in diabetes mellitus and highlights recent key descriptions of chromatinized changes associated with persistent gene expression in endothelial, smooth muscle, and circulating immune cells relevant to atherosclerosis. Furthermore, we discuss the challenges associated with pharmacological targeting of epigenetic networks to correct abnormal or deregulated gene expression as a strategy to alleviate the clinical burden of diabetic cardiovascular disease.
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    EZH2 inhibitors promote β-like cell regeneration in young and adult type 1 diabetes donors
    Al-Hasani, K ; Marikar, SN ; Kaipananickal, H ; Maxwell, S ; Okabe, J ; Khurana, I ; Karagiannis, T ; Liang, JJ ; Mariana, L ; Loudovaris, T ; Kay, T ; El-Osta, A (Nature Publishing Group, 2024-01-01)
    β-cells are a type of endocrine cell found in pancreatic islets that synthesize, store and release insulin. In type 1 diabetes (T1D), T-cells of the immune system selectively destroy the insulin-producing β-cells. Destruction of these cells leads to a lifelong dependence on exogenous insulin administration for survival. Consequently, there is an urgent need to identify novel therapies that stimulate β-cell growth and induce β-cell function. We and others have shown that pancreatic ductal progenitor cells are a promising source for regenerating β-cells for T1D owing to their inherent differentiation capacity. Default transcriptional suppression is refractory to exocrine reaction and tightly controls the regenerative potential by the EZH2 methyltransferase. In the present study, we show that transient stimulation of exocrine cells, derived from juvenile and adult T1D donors to the FDA-approved EZH2 inhibitors GSK126 and Tazemetostat (Taz) influence a phenotypic shift towards a β-like cell identity. The transition from repressed to permissive chromatin states are dependent on bivalent H3K27me3 and H3K4me3 chromatin modification. Targeting EZH2 is fundamental to β-cell regenerative potential. Reprogrammed pancreatic ductal cells exhibit insulin production and secretion in response to a physiological glucose challenge ex vivo. These pre-clinical studies underscore the potential of small molecule inhibitors as novel modulators of ductal progenitor differentiation and a promising new approach for the restoration of β-like cell function.
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    Persistent epigenetic signals propel a senescence-associated secretory phenotype and trained innate immunity in CD34+ hematopoietic stem cells from diabetic patients
    Vinci, MC ; Costantino, S ; Damiano, G ; Rurali, E ; Rinaldi, R ; Vigorelli, V ; Sforza, A ; Carulli, E ; Pirola, S ; Mastroiacovo, G ; Raucci, A ; El-Osta, A ; Paneni, F ; Pompilio, G (BMC, 2024-03-29)
    BACKGROUND: Diabetes-induced trained immunity contributes to the development of atherosclerosis and its complications. This study aimed to investigate in humans whether epigenetic signals involved in immune cell activation and inflammation are initiated in hematopoietic stem/progenitor cells (HSPCs) and transferred to differentiated progeny. METHODS AND RESULTS: High glucose (HG)-exposure of cord blood (CB)-derived HSPCs induced a senescent-associated secretory phenotype (SASP) characterized by cell proliferation lowering, ROS production, telomere shortening, up-regulation of p21 and p27genes, upregulation of NFkB-p65 transcription factor and increased secretion of the inflammatory cytokines TNFα and IL6. Chromatin immunoprecipitation assay (ChIP) of p65 promoter revealed that H3K4me1 histone mark accumulation and methyltransferase SetD7 recruitment, along with the reduction of repressive H3K9me3 histone modification, were involved in NFkB-p65 upregulation of HG-HSPCs, as confirmed by increased RNA polymerase II engagement at gene level. The differentiation of HG-HSPCs into myeloid cells generated highly responsive monocytes, mainly composed of intermediate subsets (CD14hiCD16+), that like the cells from which they derive, were characterized by SASP features and similar epigenetic patterns at the p65 promoter. The clinical relevance of our findings was confirmed in sternal BM-derived HSPCs of T2DM patients. In line with our in vitro model, T2DM HSPCs were characterized by SASP profile and SETD7 upregulation. Additionally, they generated, after myeloid differentiation, senescent monocytes mainly composed of proinflammatory intermediates (CD14hiCD16+) characterized by H3K4me1 accumulation at NFkB-p65 promoter. CONCLUSIONS: Hyperglycemia induces marked chromatin modifications in HSPCs, which, once transmitted to the cell progeny, contributes to persistent and pathogenic changes in immune cell function and composition.
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    Insulin icodec use in hospital settings: Considerations for once-weekly basal insulin therapy in hospital glycaemic management practice
    Barmanray, RD ; Kyi, M ; Rayman, G ; Rushakoff, R ; Newland-Jones, P ; Fourlanos, S (Elsevier, 2024-05)
    Diabetes management has benefitted greatly from novel insulin analogues with action profiles that better match individual’s requirements. However, the increased complexity of hospital insulin administration involving multiple practitioners, rapidly changing clinical situations, and therapies causing hyperglycaemia, demands specific consideration for their use. Insulin icodec has an extended duration of action and is beginning to be used in the ambulatory setting. A reassuring early trial experience observed no substantial dysglycaemia in 135 hospitalised participants [ 1 ]. However, the limited glucose measurements informing this observation under intensive clinical trial conditions warrants further consideration of insulin icodec’s implications for real-world hospital settings.
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    A compendium of genetic regulatory effects across pig tissues
    Teng, J ; Gao, Y ; Yin, H ; Bai, Z ; Liu, S ; Zeng, H ; Bai, L ; Cai, Z ; Zhao, B ; Li, X ; Xu, Z ; Lin, Q ; Pan, Z ; Yang, W ; Yu, X ; Guan, D ; Hou, Y ; Keel, BN ; Rohrer, GA ; Lindholm-Perry, AK ; Oliver, WT ; Ballester, M ; Crespo-Piazuelo, D ; Quintanilla, R ; Canela-Xandri, O ; Rawlik, K ; Xia, C ; Yao, Y ; Zhao, Q ; Yao, W ; Yang, L ; Li, H ; Zhang, H ; Liao, W ; Chen, T ; Karlskov-Mortensen, P ; Fredholm, M ; Amills, M ; Clop, A ; Giuffra, E ; Wu, J ; Cai, X ; Diao, S ; Pan, X ; Wei, C ; Li, J ; Cheng, H ; Wang, S ; Su, G ; Sahana, G ; Lund, MS ; Dekkers, JCM ; Kramer, L ; Tuggle, CK ; Corbett, R ; Groenen, MAM ; Madsen, O ; Godia, M ; Rocha, D ; Charles, M ; Li, C-J ; Pausch, H ; Hu, X ; Frantz, L ; Luo, Y ; Lin, L ; Zhou, Z ; Zhang, Z ; Chen, Z ; Cui, L ; Xiang, R ; Shen, X ; Li, P ; Huang, R ; Tang, G ; Li, M ; Zhao, Y ; Yi, G ; Tang, Z ; Jiang, J ; Zhao, F ; Yuan, X ; Liu, X ; Chen, Y ; Xu, X ; Zhao, S ; Zhao, P ; Haley, C ; Zhou, H ; Wang, Q ; Pan, Y ; Ding, X ; Ma, L ; Li, J ; Navarro, P ; Zhang, Q ; Li, B ; Tenesa, A ; Li, K ; Liu, GE ; Zhang, Z ; Fang, L (NATURE PORTFOLIO, 2024-01)
    The Farm Animal Genotype-Tissue Expression (FarmGTEx) project has been established to develop a public resource of genetic regulatory variants in livestock, which is essential for linking genetic polymorphisms to variation in phenotypes, helping fundamental biological discovery and exploitation in animal breeding and human biomedicine. Here we show results from the pilot phase of PigGTEx by processing 5,457 RNA-sequencing and 1,602 whole-genome sequencing samples passing quality control from pigs. We build a pig genotype imputation panel and associate millions of genetic variants with five types of transcriptomic phenotypes in 34 tissues. We evaluate tissue specificity of regulatory effects and elucidate molecular mechanisms of their action using multi-omics data. Leveraging this resource, we decipher regulatory mechanisms underlying 207 pig complex phenotypes and demonstrate the similarity of pigs to humans in gene expression and the genetic regulation behind complex phenotypes, supporting the importance of pigs as a human biomedical model.
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    Evolution and transmission of antibiotic resistance is driven by Beijing lineage Mycobacterium tuberculosisin Vietnam
    Silcocks, M ; Chang, X ; Thuong, NTT ; Qin, Y ; Ha, DTM ; Thai, PVK ; Vijay, S ; Thu, DDA ; Ha, VTN ; Nhung, HN ; Lan, NH ; Nhu, NTQ ; Edwards, D ; Nath, A ; Pham, K ; Bang, ND ; Chau, TTH ; Thwaites, G ; Heemskerk, AD ; Chuen Khor, C ; Teo, YY ; Inouye, M ; Ong, RT-H ; Caws, M ; Holt, KE ; Dunstan, SJ ; Neyrolles, O (AMER SOC MICROBIOLOGY, 2023-12-12)
    Drug-resistant tuberculosis (TB) infection is a growing and potent concern, and combating it will be necessary to achieve the WHO's goal of a 95% reduction in TB deaths by 2035. While prior studies have explored the evolution and spread of drug resistance, we still lack a clear understanding of the fitness costs (if any) imposed by resistance-conferring mutations and the role that Mtb genetic lineage plays in determining the likelihood of resistance evolution. This study offers insight into these questions by assessing the dynamics of resistance evolution in a high-burden Southeast Asian setting with a diverse lineage composition. It demonstrates that there are clear lineage-specific differences in the dynamics of resistance acquisition and transmission and shows that different lineages evolve resistance via characteristic mutational pathways.