Human Induced Pluripotent Stem Cell as a Disease Modeling and Drug Development Platform—A Cardiac Perspective

Abstract

A comprehensive understanding of the pathophysiology and cellular responses to drugs in human heart disease is limited by species differences between humans and experimental animals. In addition, isolation of human cardiomyocytes (CMs) is complicated because cells obtained by biopsy do not proliferate to provide sufficient numbers of cells for preclinical studies in vitro. Interestingly, the discovery of human-induced pluripotent stem cell (hiPSC) has opened up the possibility of generating and studying heart disease in a culture dish. The combination of reprogramming and genome editing technologies to generate a broad spectrum of human heart diseases in vitro offers a great opportunity to elucidate gene function and mechanisms. However, to exploit the potential applications of hiPSC-derived-CMs for drug testing and studying adult-onset cardiac disease, a full functional characterization of maturation and metabolic traits is required. In this review, we focus on methods to reprogram somatic cells into hiPSC and the solutions for overcome immaturity of the hiPSC-derived-CMs to mimic the structure and physiological properties of the adult human CMs to accurately model disease and test drug safety. Finally, we discuss how to improve the culture, differentiation, and purification of CMs to obtain sufficient numbers of desired types of hiPSC-derived-CMs for disease modeling and drug development platform.

Human induced pluripotent stem cell-derived cardiac myocytes and sympathetic neurons in disease modelling

“…Briefly, after three passages, hiPSC were seeded on Matrigel-coated 12-well plates and cultured in mTeSR until 80% confluent. Then a series of chemical compounds were added sequentially (500 nM LDN193189 (Sigma, SML0559) days 0-3, 10 µM SB431542 (Sigma, S4317) days 0-4, 3 µM CHIR99021 + 10 µM DAPT (Sigma, D5942) + 0.2 µM PD173074 (Sigma, P2499) days 2-7, 60 ng ml −1 Shh C25II (Gibco, PMC8034) + 1 µM PMP (Sigma, SML0868) days 3-12, 10 ng ml −1 BMP4 (R&D Systems, 314-BP-010) days [10][11][12][13][14] to the culture medium to modify pathways involved in the induction of neuromesodermal progenitors and neural crest cells. Culture medium mTeSR was gradually replaced by N2 medium (Neurobasal Plus medium (Gibco, A3582901) supplemented with 2 mM L-glutamine, B-27 Plus (Gibco, A3582801), N-2 supplements (Gibco, 11520536), 0.2 mM ascorbic acid (Sigma, A8960), 0.2 nM dbcAMP (Sigma, D0260), 10 ng ml −1 NGF (Bio-Techne, 256-GF-100), 10 ng ml −1 BDNF (R&D Systems, 248-BDB-010), 10 ng ml −1 GDNF (R&D Systems, 212-GD-010)) to promote sympathetic neuron development from day 4 (days 4-5: 75% mTeSR + 25% N2 medium; days 6-7: 50% mTeSR + 50% N2 medium; days 8-9: 25% mTeSR + 75% N2 medium; days 10-12: N2 medium).…”

“…However, it was the reprogramming of fibroblasts back to their pluripotent state in 2006 by Takahashi & Yamanaka that resulted in human induced pluripotent stem cell technology transforming biology and providing an opportunity to study human disease in vitro [3,4]. In heart physiology, the induction of cardiomyocytes from human induced pluripotent stem cells (hiPSC) was rapidly realized by applying the standard embryoid bodies-based protocol for embryonic stem cells [5,6], which was optimized over time by more precise control of the embryonic development signal [7][8][9][10]. To date, the speed to generate hiPSC has been enhanced and genetic stability improved using novel modified synthetic mRNA transfection methods [11,12].…”

Deciphering Common Long QT Syndrome Using CRISPR/Cas9 in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes

“…Circumventing the ethical limitations of hESCs, reprogrammed pluripotent stem cells from somatic cells of LQTS patients are induced by four specific transcription factors (28)(29)(30). Transcription factors are mainly delivered into any cell type with proliferative potential by viral vector-based methods (retroviruses, lentiviruses, and inducible viruses) and non-viral vector methods (plasmids or linear DNA and transposons) (31). Initially, skin dermal fibroblasts were used, followed by non-invasive peripheral blood cells, including T and B cells, and even uroepithelial cells (32,33).…”

High-throughput optical action potential recordings in hiPSC-derived cardiomyocytes with a genetically encoded voltage indicator in the AAVS1 locus

“…In the past decade, human induced pluripotent stem cellderived cardiomyocytes (hiPSC-CMs) have opened unprecedented possibilities to investigate mechanisms of human cardiovascular diseases (Moretti et al, 2010;Dorn et al, 2018;Bekhite and Schulze, 2021;Funakoshi and Yoshida, 2021;Krane et al, 2021;Micheu and Rosca, 2021;Meier et al, 2022) and to explore novel treatment options (Gramlich et al, 2015;Csobonyeiova et al, 2016;Gharanei et al, 2022). They have also served as a safety pharmacology platform to assess drug-induced cardiotoxicity, including lifethreatening proarrhythmia, a major concern in pharmaceutical development (Gintant et al, 2017).…”