Multiomics-based assessment of 2D and 3D human iPSC-cardiomyocyte models of insulin resistance demonstrate metabolic and contractile dysfunction that recapitulates diabetic cardiomyopathy.

Abstract

In type II diabetes (T2DM), the heart is exposed to hyperglycaemia, hyperlipidaemia, and hyperinsulinaemia, leading to insulin resistance and metabolic dysfunction, culminating in diabetic cardiomyopathy (DbCM). Human-centric models of DbCM are needed to provide mechanistic insights and therapeutic targets in a translationally relevant setting. We hypothesised that culturing human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in an "insulin resistance" (IR) media, and assessing this using a systems biology approach, would offer a comprehensive evaluation of dysregulated pathways, establishing their suitability as a model of DbCM. Culturing hiPSC-CMs in 2D or 3D as engineered heart tissue (EHT) in IR media induced insulin resistance and activated numerous pathways implicated in DbCM, including metabolic remodelling, mitochondrial dysfunction, extracellular matrix remodelling, and endoplasmic reticulum stress. Pathways involved in fatty acid oxidation were upregulated, while those involved in glucose metabolism were downregulated, which was validated using radioisotope flux measurements. Adaptation to hypoxia, a key component of post-ischaemic remodelling, was blunted in the 2D IR hiPSC-CMs. Combining proteomic and transcriptomic analyses in the IR 3D EHT revealed significant enrichment of DbCM pathways, with subnetworks enriched for several metabolic and diabetes-related pathways. Additionally, IR 3D EHT displayed impaired relaxation, mimicking the diastolic dysfunction observed in T2DM patients. In conclusion, culturing hiPSC-CM in 2D or 3D in an IR media activates multiple mechanisms implicated in the development of DbCM, with IR 3D EHT also recapitulating the diastolic dysfunction present in patients with T2DM.