Neuregulin-1β Induces Mature Ventricular Cardiac Differentiation from Induced Pluripotent Stem Cells Contributing to Cardiac Tissue Repair

Abstract

Stem cell-derived cardiomyocytes (CMs) are often electrophysiologically immature and heterogeneous, which represents a major barrier to their in vitro and in vivo application. Therefore, the purpose of this study was to examine whether Neuregulin-1b (NRG-1b) treatment could enhance in vitro generation of mature ''workingtype'' CMs from induced pluripotent stem (iPS) cells and assess the regenerative effects of these CMs on cardiac tissue after acute myocardial infarction (AMI). With that purpose, adult mouse fibroblast-derived iPS from a-MHC-GFP mice were derived and differentiated into CMs through NRG-1b and/or dimethyl sulfoxide (DMSO) treatment. Cardiac specification and maturation of the iPS was analyzed by gene expression array, quantitative real-time polymerase chain reaction, immunofluorescence, electron microscopy, and patch-clamp techniques. In vivo, the iPS-derived CMs or culture medium control were injected into the peri-infarct region of hearts after coronary artery ligation, and functional and histology changes were assessed from 1 to 8 weeks post-transplantation. On differentiation, the iPS displayed early and robust in vitro cardiogenesis, expressing cardiac-specific genes and proteins. More importantly, electrophysiological studies demonstrated that a more mature ventricular-like cardiac phenotype was achieved when cells were treated with NRG-1b and DMSO compared with DMSO alone. Furthermore, in vivo studies demonstrated that iPS-derived CMs were able to engraft and electromechanically couple to heart tissue, ultimately preserving cardiac function and inducing adequate heart tissue remodeling. In conclusion, we have demonstrated that combined treatment with NRG-1b and DMSO leads to efficient differentiation of iPS into ventricular-like cardiac cells with a higher degree of maturation, which are capable of preserving cardiac function and tissue viability when transplanted into a mouse model of AMI.

Human pluripotent stem cells: Prospects and challenges as a source of cardiomyocytes for in vitro modeling and cell-based cardiac repair

“…Of note, T3 is present in the serum substitute, B-27 (Life Technologies), which is used as a media constituent in multiple directed cardiomyocyte differentiation protocols. Other factors have been reported to promote the maturation of at least some parameters in cardiomyocytes from mouse or human PSCs include neuregulin [119, 120], angiotensin II [63], and the alpha-adrenergic agonist phenylephrine [63]. Finally, while it has been suggested that non-myocytes secrete as yet undefined factors that favor human PSC-derived cardiomyocyte maturation [61], our own data demonstrate that these factors are not absolutely required for maturation [60].…”

Towards the standardization of stem cell therapy studies for ischemic heart diseases: Bridging the gap between animal models and the clinical setting

“…The advantages and limitations of each stem cell with regard to the possible emergence of ethical concerns (symbolized by ¥), the invasiveness and collection (from + to ++++), the isolation procedure, the abundance (from + to ++++), expansion and proliferation rate, the transdifferentiation capacity, the paracrine effects or/and cardiomyogenic potential, the survival and engraftment capacity as well as the immunogenic, oncogenic or teratogenic risks is given in the accessory side tables. The left table abridges bone marrow-derived stem cells [3,[19][20][21]27,28], cortical bone-derived stem cells [32,33], adipose tissue-derived stem cells [34,37,39,41] and gestation-related stem cells [42][43][44][45][46][47][48][49]51,54] and the right table abridges cardiac stem cells [57,59,60], skeletal muscle stem cells [62,63,66] and induced pluripotent stem cells [67,[69][70][71]. The human body scheme was constructed with images sourced from Servier Medical Art (http://www.servier.com/ Powerpoint-image-bank) and Cientic website (http://www.cientic.com/).…”

“…Virtually, all adult cells can be subjected to a dedifferentiation program with a set of stemness factors to become induced pluripotent stem cells (iPSC) [67][68][69]. If not collected from an autologous source, iPSC are related to immunogenic and teratogenic risks (Fig.…”

“…If not collected from an autologous source, iPSC are related to immunogenic and teratogenic risks (Fig. 1), though these cells are readily available, as their progenitors are somatic cells, bypassing the ethical concerns related to the collection of ESC [67][68][69][70]. Again, paracrine activity is believed to be the main mechanism of iPSC action through, for instance, angiogenesis stimulation [71].…”

Novel Biological Therapies Targeting Heart Failure

“…The NRG-1β isoform of NRG has effects on cardiomyocyte survival as well as structure and function in vitro, in addition to endothelial cell and fibroblast responses 42,43 . In addition, NRG-1β activates differentiation of progenitor cell populations toward cardiac cell lineages, at least in vitro 44,45 . Some or all of these effects may explain how recombinant NRG-1β given to both animals and humans with systolic dysfunction is associated with improvements in heart function.…”