IGF1 and NRG1 Enhance Proliferation, Metabolic Maturity, and the Force-Frequency Response in hESC-Derived Engineered Cardiac Tissues

Abstract

Insulin-like growth factor 1 (IGF1) and neuregulin-1β (NRG1) play important roles during cardiac development both individually and synergistically. In this study, we analyze how 3D cardiac tissue engineered from human embryonic stem cell- (hESC-) derived cardiomyocytes and 2D-plated hESC-cardiomyocytes respond to developmentally relevant growth factors both to stimulate maturity and to characterize the therapeutic potential of IGF1 and NRG1. When administered to engineered cardiac tissues, a significant decrease in active force production of ~65% was measured in all treatment groups, likely due to changes in cellular physiology. Developmentally related processes were identified in engineered tissues as IGF1 increased hESC-cardiomyocyte proliferation 3-fold over untreated controls and NRG1 stimulated oxidative phosphorylation and promoted a positive force-frequency relationship in tissues up to 3 Hz. hESC-cardiomyocyte area increased significantly with NRG1 and IGF1 + NRG1 treatment in 2D culture and gene expression data suggested increased cardiac contractile components in engineered tissues, indicating the need for functional analysis in a 3D platform to accurately characterize engineered cardiac tissue response to biochemical stimulation. This study demonstrates the therapeutic potential of IGF1 for boosting proliferation and NRG1 for promoting metabolic and contractile maturation in engineered human cardiac tissue.

Practical adoption of state-of-the-art hiPSC-cardiomyocyte differentiation techniques

“…Engineered cardiac tissues (ECTs) were formed as previously described. [15] Briefly, hiPSCcardiomyocytes were harvested at day 24 of differentiation and combined with 1.25 mg/mL rat-tail collagen-1 at 1x10 6 cells per tissue + 5% hCFs. ECTs were cultured under static stress [16] in RPMI/B27 with 1% penicillin-streptomycin and field-stimulated at a 1 Hz (IonOptix C-pace EP).…”

“…Engineered cardiac tissues (ECTs) underwent mechanical analysis after one week of culture as previously described [15]. Tissues were mounted on one side to a 5 mN load cell and on the other to a high-speed length controller (Aurora Scientific).…”

Regulation of cardiomyocyte maturation during critical perinatal window

“…Similarly, in rodents, thyroid hormone secretion initiates at approximately e17.5, and mediates transcriptional effects on CM contractile function, among other effects . Other hormones that have been implicated in CM maturation include IGF1 and NRG1 (Rupert & Coulombe, 2017).…”

The Role of Reactive Oxygen Species in In Vitro Cardiac Maturation

“…The employed methods included: (i) mechanical stimulation, such as passive stretching of the cells [49] and dynamic culture on shakers [7], and (ii) electrical stimulation with biowires [4] or, more recently, the application of a high-frequency electrical field [8]. Additionally, (iii) biochemical stimulation, such as insulin-like growth factor-1 (IGF1), neuregulin-1ß [50], and Tri-iodo-l-thyronine 3 (T3); [51], and (iv) metabolic modulation, such as depletion of glucose [52] or use of galactose and fatty acids (FAs)-containing medium [2,6] have been added to cardiac maintenance medium to encourage further maturation. Despite these notable advances, improvements in hPSC-CM maturity have remained mostly relative and partial.…”