Comparison of 10 Control hPSC Lines for Drug Screening in an Engineered Heart Tissue Format

Mannhardt, Ingra; Saleem, Umber; Mosqueira, Diogo; Loos, Malte; Ulmer, Bärbel; Lemoine, Marc D.; Larsson, Camilla; Améen, Caroline; Korte, Tessa de; Vlaming, Maria L. H.; Harris, Kate; Clements, Peter; Denning, Chris; Hansen, Arne; Eschenhagen, Thomas

doi:10.1016/j.stemcr.2020.09.002

Cited by 53 publications

(44 citation statements)

References 55 publications

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“…To vary the dosage of the applied cardiomyocyte number, we generated EHTs with different cell densities (cell-free, fibrin patches without cells; dose 1, 9×10 6 cells; dose 2, 12×10 6 cells; and dose 3, 15×10 6 cells per 1.5 mL). Because cell number influences EHT development, 17 it was not possible to only decrease cardiomyocyte number per patch. We therefore used a strategy combining cell number and patch area to create patches with different cell content.…”

Section: Methodsmentioning

confidence: 99%

Human Engineered Heart Tissue Patches Remuscularize the Injured Heart in a Dose-Dependent Manner

et al. 2021

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Background: Human engineered heart tissue (EHT) transplantation represents a potential regenerative strategy for heart failure patients and has been successful in preclinical models. Clinical application requires upscaling, adaptation to good manufacturing practices (GMP) and determination of the effective dose. Methods: Cardiomyocytes were differentiated from three different human induced pluripotent stem cell (hiPSC) lines including one reprogrammed under GMP conditions. Protocols for hiPSC expansion, cardiomyocyte differentiation and EHT generation were adapted to substances available in GMP quality. EHT geometry was modified to generate patches suitable for transplantation in a small animal model and perspectively humans. Repair efficacy was evaluated at 3 doses in a cryo-injury guinea pig model. Human-scale patches were epicardially transplanted onto healthy hearts in pigs to assess technical feasibility. Results: We created mesh structured tissue patches for transplantation in guinea pigs (1.5x2.5 cm, 9-15x10 6 cardiomyocytes) and pigs (5x7 cm, 450 x10 6 cardiomyocytes). EHT patches coherently beat in culture and developed high force (mean 4.6 mN). Cardiomyocytes matured, aligned along the force lines, and demonstrated advanced sarcomeric structure and action potential characteristics closely resembling human ventricular tissue. EHT patches containing ~4.5, 8.5, 12x10 6 or no cells were transplanted 7 days after cryo-injury (n=18-19 per group). EHT transplantation resulted in a dose-dependent remuscularization (graft size: 0-12% of the scar). Only high-dose patches improved left-ventricular function (+8% absolute, +24% relative increase). The grafts showed time-dependent cardiomyocyte proliferation. While standard EHT patches did not withstand transplantation in pigs, the human-scale patch enabled successful patch transplantation. Conclusions: EHT patch transplantation resulted in a partial remuscularization of the injured heart and improved left-ventricular function in a dose-dependent manner in a guinea pig injury model. Human scale patches were successfully transplanted in pigs in a proof-of-principle study.

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Section: Methodsmentioning

confidence: 99%

Human Engineered Heart Tissue Patches Remuscularize the Injured Heart in a Dose-Dependent Manner

et al. 2021

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“…Routine assessments of hPSC-CM combine analyses at the transcript, protein, and metabolite level with phenotypic and physiological recordings [10,28,41,59,82,108]. However, neither glycan structures in general, nor the presence of a specific glycan structure at a specific glycosite within a protein, can be directly predicted from transcriptomic, proteomic, or metabolomic studies.…”

Section: Strategies To Assess Protein Glycosylationmentioning

confidence: 99%

Importance of evaluating protein glycosylation in pluripotent stem cell-derived cardiomyocytes for research and clinical applications

Kelly

Albahrani

Castro

et al. 2021

Pflugers Arch - Eur J Physiol

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Proper protein glycosylation is critical to normal cardiomyocyte physiology. Aberrant glycosylation can alter protein localization, structure, drug interactions, and cellular function. The in vitro differentiation of human pluripotent stem cells into cardiomyocytes (hPSC-CM) has become increasingly important to the study of protein function and to the fields of cardiac disease modeling, drug testing, drug discovery, and regenerative medicine. Here, we offer our perspective on the importance of protein glycosylation in hPSC-CM. Protein glycosylation is dynamic in hPSC-CM, but the timing and extent of glycosylation are still poorly defined. We provide new data highlighting how observed changes in hPSC-CM glycosylation may be caused by underlying differences in the protein or transcript abundance of enzymes involved in building and trimming the glycan structures or glycoprotein gene products. We also provide evidence that alternative splicing results in altered sites of glycosylation within the protein sequence. Our findings suggest the need to precisely define protein glycosylation events that may have a critical impact on the function and maturation state of hPSC-CM. Finally, we provide an overview of analytical strategies available for studying protein glycosylation and identify opportunities for the development of new bioinformatic approaches to integrate diverse protein glycosylation data types. We predict that these tools will promote the accurate assessment of protein glycosylation in future studies of hPSC-CM that will ultimately be of significant experimental and clinical benefit.

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“…One could potentially expect different responses to the drugs investigated based on racial/ethnic background as well as genetic variants (pharmacogenomics). In this context, it is noteworthy that a recent drug screening report of 10 control human pluripotent stem cell lines in an engineered heart tissue format found that, while there was great variability in baseline contractile parameters across the different lines, the variability appeared less relevant for drug screening ( Mannhardt et al., 2020 ). A number of constraints precluded us from testing every hiPSC line in our library for each of the conditions and assays.…”

Section: Discussionmentioning

confidence: 99%

A library of induced pluripotent stem cells from clinically well-characterized, diverse healthy human individuals

Schaniel

Dhanan

et al. 2021

Stem Cell Reports

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Summary A library of well-characterized human induced pluripotent stem cell (hiPSC) lines from clinically healthy human subjects could serve as a useful resource of normal controls for in vitro human development, disease modeling, genotype-phenotype association studies, and drug response evaluation. We report generation and extensive characterization of a gender-balanced, racially/ethnically diverse library of hiPSC lines from 40 clinically healthy human individuals who range in age from 22 to 61 years. The hiPSCs match the karyotype and short tandem repeat identities of their parental fibroblasts, and have a transcription profile characteristic of pluripotent stem cells. We provide whole-genome sequencing data for one hiPSC clone from each individual, genomic ancestry determination, and analysis of mendelian disease genes and risks. We document similar transcriptomic profiles, single-cell RNA-sequencing-derived cell clusters, and physiology of cardiomyocytes differentiated from multiple independent hiPSC lines. This extensive characterization makes this hiPSC library a valuable resource for many studies on human biology.

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Comparison of 10 Control hPSC Lines for Drug Screening in an Engineered Heart Tissue Format

Cited by 53 publications

References 55 publications

Human Engineered Heart Tissue Patches Remuscularize the Injured Heart in a Dose-Dependent Manner

Human Engineered Heart Tissue Patches Remuscularize the Injured Heart in a Dose-Dependent Manner

Importance of evaluating protein glycosylation in pluripotent stem cell-derived cardiomyocytes for research and clinical applications

A library of induced pluripotent stem cells from clinically well-characterized, diverse healthy human individuals

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