Development of matured hiPSCs-derived 3D cardiac tissue using ERR gamma agonist and mechanical stress and application for Hypertrophic Cardiomyopathy (HCM) model

Abstract

Introduction Tissue engineering using human induced pluripotent stem cells-derived cardiomyocytes (hiPSCs-CMs) is one of the potential tools to replicate human heart in vitro. Although there are many publications on 3 dimensional (3D) heart tissues (1), these tissues show fetal like phenotypes. For that reason, several maturation methods such as electrical stimulation and mechanical stress have been investigated (2, 3). However, these methods have been inadequate in differentiating fetal like phenotype tissue from adult tissues. Previously, we identified a novel compound, T112, which induced hiPSCs-CMs maturation from approximately 9,000 compounds using Troponin I1-EmGFP and Troponin I3-mCherry double reporter hiPSCs-CMs. This compound enhanced morphological and metabolic maturation of hiPSCs-CMs via estrogen-rerated receptor gamma activation Purpose We hypothesized that our novel compound, T112, in combination with mechanical stress could result in further maturation of 3D heart tissue. Therefore, our specific aim is to develop a novel maturation method applicable to genetic disease model of HCM using 3D heart tissue combined with T112. Methods We constructed 3D heart tissue mixed with fibroblast and double reporter hiPSCs-CMs by the hydrogel methods using Flex cell system®. We added T112 with or without mechanical stretching to 3D tissue from 7 to 15 days after 3D heart tissue was constructed. Then we measured maturation related phenotype such as sarcomere gene expression, mitochondrial DNA content and cell size. Results Similar to hiPSCs-CM, the addition of T112 to the constructed 3D heart tissue significantly increased TNNI3 mRNA compared to that of DMSO. Furthermore, T112 treated 3D heart tissue showed increased cell size and oblong shape. Next, in order to promote more maturation of 3D heart tissue, we performed mechanical stretching with the addition of T112. The combination of T112 with mechanical stretching showed higher expression of mCherry, a reporter protein for TNNI3 expression, and higher isotropy of sarcomere alignment in 3D heart tissue than that with the static condition. Furthermore, 3D heart tissue in the treatment of T112 with or without mechanical stretching showed higher mitochondrial DNA content compared to the respective DMSO controls. Interestingly, we applied this combination method to hiPSCs carrying MYH7 R719Q mutation which is known to cause hypertrophic cardiomyopathy, and the 3D heart tissue composed of cardiomyocytes derived from mutant iPSCs demonstrated increased sarcomere disarray compared to isogenic wild-type 3D heart tissue. Conclusion These results suggest that the combination of T112 and mechanical stretching promotes metabolic and structural maturation of 3D heart tissue and would be useful for creating a HCM disease model. Funding Acknowledgement Type of funding source: Private company. Main funding source(s): T-CiRA project, Takeda Pharmaceutical Company Limited

Novel Pan-ERR Agonists Ameliorate Heart Failure Through Enhancing Cardiac Fatty Acid Metabolism and Mitochondrial Function

“…The essential role of ERRα and ERRγ in controlling cardiac metabolism and mitochondrial and contractile function has been demonstrated by early studies of genetic knockout mouse models [9][10][11][12][13] ; however, the ability to activate these processes in vivo with ERR agonists has not been demonstrated previously. Although chemical library screening has led to identification of agonists for ERRα, 17,18 ERRγ, 19,93 and ERRβ/γ, 20 no study has demonstrated the bioavailability of these compounds for in vivo use. As such, it was necessary for us to develop a novel chemical tool compound that would have in vivo exposure so that the therapeutic potential of targeting ERRs in vivo could be evaluated.…”

“…14 Therefore, ERRs have been proposed as candidate therapeutic targets for treating HF and other metabolic diseases. 15,16 Although several ERR agonists have been developed, [17][18][19][20] no study has demonstrated their potential use for HF treatment in vivo. As such, the therapeutic potential of these critical metabolic regulators remains untested.…”

Novel ERR pan-agonists ameliorate heart failure through boosting cardiac fatty acid metabolism and mitochondrial function

“…While ERR-targeting therapy has been proposed for heart failure therapy, no natural ligand was identified nor synthetic agonist with favorable pharmacokinetics that can be used for in vivo studies had been developed (15, 16). Although there have been compounds identified as ERRα agonists (17, 18), ERRγ agonists (19), and ERRβ/γ agonists (20) through chemical library screening, none of them have been available for in vivo use and examined as a therapeutic agent for heart failure treatment in vivo . We believe that it was necessary to develop a chemical “tool” compound that would have in vivo exposure so that the therapeutic potential of targeting ERRs in vivo could be evaluated.…”

“…Therefore, ERRs have been proposed as candidate therapeutic targets for treating heart failure and other metabolic diseases (15,16). Despite several ERR agonists having been previously developed (17)(18)(19)(20), no study has demonstrated the potential utility for heart failure treatment in vivo. In this study, we report the cardiac protective effect of a recently described ERR pan-agonist SLU-PP-332 (332) and another novel structurally distinct ERR pan-agonist SLU-PP-915 (915) in pressure overload-induced heart failure.…”