谷 英典

The prognosis for heart failure (HF) with preserved ejection fraction (HFpEF) remains poor, with treatment evidence and studies at the human cellular level limited. Here, we aimed to model HFpEF-associated diastolic dysfunction in vitro by generating human engineered heart tissues (hEHTs) using human induced pluripotent stem cells and culturing the tissues under high fatty acid and L-NG-nitroarginine methyl ester supplementation. Medium-loaded hEHTs showed a marked reduction in relaxation function while preserving contraction function; secreted high levels of the HF marker, BNP; exhibited abnormal calcium transients; and showed structural and functional features of HF. After treatment with several existing HF drugs, a sodium-glucose cotransporter 2 inhibitor (SGLT2i) improved the decline in relaxation function and contributed to an improvement in the diastolic dysfunction phenotype. This mechanism exhibited an anti-inflammatory effect mediated by the recovery of the eNOS-NO-cGMP-PKG signaling pathway. These findings serve as a basis for elucidating the pathogenesis and mechanisms of improvement in HFpEF.

Human induced pluripotent stem cells (hiPSCs) have emerged as a promising platform for elucidating disease mechanisms and developing new drugs. Over the past 2 decades, it has become possible to efficiently generate large quantities of cardiomyocytes (CMs) from hiPSCs, thereby enabling the reproduction of disease-specific characteristics in culture dishes. Although this technology has the potential to substantially enhance the efficiency of drug discovery and understanding of disease, the immaturity of hiPSC-derived CMs (hiPSC-CMs) has been a major barrier to their widespread adoption. This review discusses the recent advances that address these challenges and explores the potential of hiPSCs to advance disease modeling, elucidate disease mechanisms, and accelerate drug discovery.

Human pluripotent stem cell-derived cardiomyocyte (hPSC-CM) differentiation can improve using chemical compounds which mimic early heart development. However, variations in hPSC-CM differentiation efficiency and its poor reproducibility have remained a challenge. Here, we report a unique metabolic method to promote hPSC-CM differentiation that involves marked suppression of the mitochondrial oxidative phosphorylation from the mesendoderm to the cardiac mesoderm, which is regulated by PHGDH, a rate-limiting enzyme in the serine synthesis pathway. Mechanistically, PHGDH inhibition impairs mitochondrial respiration by blocking the electron transport chain, resulting in elevated ROS levels and promoting the cardiomyocyte lineage specification by disrupting the cardiopharyngeal mesoderm lineage differentiation. Additionally, antioxidant supplementation can scavenge ROS and eliminate the effects of PHGDH inhibition. Collectively, our findings show that serine synthesis pathway can regulate cardiomyocyte lineage specification and have implications in providing a cellular source for transplantation and elucidating the potential mechanisms of heart development and pathogenesis of heart disease.

Three-dimensional cultures mimic in vivo environments better than two-dimensional cultures and are often used in drug discovery research. Herein, we present a protocol for producing homogeneous induced pluripotent stem cell (iPSC) spheroids and microtissues using the suction technique. We describe steps for preparing the suction device, preparing and seeding cells, and suction sedimentation of cells. We then detail procedures for self-assembly and evaluation of spheroids. For complete details on the use and execution of this protocol, please refer to Moriwaki et al.1.