Human induced pluripotent stem cells for studying mitochondrial diseases in the heart

Caudal, Arianne; Ren, Lu; Tu, Chunling; Wu, Joseph C.

doi:10.1002/1873-3468.14444

Cited by 10 publications

(5 citation statements)

References 95 publications

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“…In the last decade, with the booming number of congenital and acquired metabolic disorders identified [71], the need to develop in vitro disease models via patient-derived hiPSCs to study pathogenic mechanisms and screen drugs preclinically is widely increasing. In the field of nucleotide sugar biosynthesis defects, to date only few hiPSC-based studies have been reported, specifically targeting phosphomannomutase 2 (PMM2) deficiency and UDP-glucose pyrophosphorylase (UGP2) deficiency [72][73][74], while they are more extensively used to model other inborn errors of metabolism, such as Pompe disease or mitochondrial disorders [75][76][77][78][79][80][81][82]. Nonetheless, despite the rapidly increasing interest in hiPSCs and hiPSC-derived models to study metabolic diseases, dynamic studies of metabolic pathways via isotopic labeling remains mostly unexplored.…”

Section: Discussionmentioning

confidence: 99%

Isotopic Tracing of Nucleotide Sugar Metabolism in Human Pluripotent Stem Cells

Conte,

Noga,

van Scherpenzeel

et al. 2023

Cells

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Metabolism not only produces energy necessary for the cell but is also a key regulator of several cellular functions, including pluripotency and self-renewal. Nucleotide sugars (NSs) are activated sugars that link glucose metabolism with cellular functions via protein N-glycosylation and O-GlcNAcylation. Thus, understanding how different metabolic pathways converge in the synthesis of NSs is critical to explore new opportunities for metabolic interference and modulation of stem cell functions. Tracer-based metabolomics is suited for this challenge, however chemically-defined, customizable media for stem cell culture in which nutrients can be replaced with isotopically labeled analogs are scarcely available. Here, we established a customizable flux-conditioned E8 (FC-E8) medium that enables stem cell culture with stable isotopes for metabolic tracing, and a dedicated liquid chromatography mass-spectrometry (LC-MS/MS) method targeting metabolic pathways converging in NS biosynthesis. By 13C6-glucose feeding, we successfully traced the time-course of carbon incorporation into NSs directly via glucose, and indirectly via other pathways, such as glycolysis and pentose phosphate pathways, in induced pluripotent stem cells (hiPSCs) and embryonic stem cells. Then, we applied these tools to investigate the NS biosynthesis in hiPSC lines from a patient affected by deficiency of phosphoglucomutase 1 (PGM1), an enzyme regulating the synthesis of the two most abundant NSs, UDP-glucose and UDP-galactose.

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Section: Discussionmentioning

confidence: 99%

Isotopic Tracing of Nucleotide Sugar Metabolism in Human Pluripotent Stem Cells

Conte,

Noga,

van Scherpenzeel

et al. 2023

Cells

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“…Previous evidence indicates PGC-1α as a central regulator of cardiac metabolism and promoter of oxidative phosphorylation at the expense of glycolysis [77,78]. Whereas AMPK upregulation leads to improved mitochondrial activity and homeostasis in cardiomyocytes [79][80][81][82]. We found that AA-treated iPSC-CMs have undergone significant metabolic changes; genes responsible for the electron transport chain, oxidative phosphorylation, and fatty acid β-oxidation in mitochondria (NDUFV3, COX3, and COX5B) were highly expressed (Additional file 1: Fig.…”

Section: Discussionmentioning

confidence: 99%

Small molecule-mediated rapid maturation of human induced pluripotent stem cell-derived cardiomyocytes

et al. 2022

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Background Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs) do not display all hallmarks of mature primary cardiomyocytes, especially the ability to use fatty acids (FA) as an energy source, containing high mitochondrial mass, presenting binucleation and increased DNA content per nuclei (polyploidism), and synchronized electrical conduction. This immaturity represents a bottleneck to their application in (1) disease modelling—as most cardiac (genetic) diseases have a middle-age onset—and (2) clinically relevant models, where integration and functional coupling are key. So far, several methods have been reported to enhance iPSC-CM maturation; however, these protocols are laborious, costly, and not easily scalable. Therefore, we developed a simple, low-cost, and rapid protocol to promote cardiomyocyte maturation using two small molecule activators of the peroxisome proliferator-activated receptor β/δ and gamma coactivator 1-alpha (PPAR/PGC-1α) pathway: asiatic acid (AA) and GW501516 (GW). Methods and Results Monolayers of iPSC-CMs were incubated with AA or GW every other day for ten days resulting in increased expression of FA metabolism-related genes and markers for mitochondrial activity. AA-treated iPSC-CMs responsiveness to the mitochondrial respiratory chain inhibitors increased and exhibited higher flexibility in substrate utilization. Additionally, structural maturity improved after treatment as demonstrated by an increase in mRNA expression of sarcomeric-related genes and higher nuclear polyploidy in AA-treated samples. Furthermore, treatment led to increased ion channel gene expression and protein levels. Conclusions Collectively, we developed a fast, easy, and economical method to induce iPSC-CMs maturation via PPAR/PGC-1α activation. Treatment with AA or GW led to increased metabolic, structural, functional, and electrophysiological maturation, evaluated using a multiparametric quality assessment.

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“…Prominent examples include the Barth syndrome (BTHS) and Friedreich’s ataxia syndrome (FRDA). An overall list of mtDNA and nucDNA genes contributing to mitochondrial function and corresponding mutations in a disease context have been extensively reported elsewhere ( El-Hattab and Scaglia, 2016 ; Barca et al, 2020 ; Pavez-Giani and Cyganek, 2021 ; Caudal et al, 2022 ). Since these syndromes are based on nuclear or mitochondrial gene mutations with a prominent cardiac phenotype, the iPSC-CM platform provides an excellent tool to study the underlying disease drivers.…”

Section: Insights Into Mitochondrial Cardiomyopathies Using Ipsc-cmmentioning

confidence: 99%

How can we use stem cell-derived cardiomyocytes to understand the involvement of energetic metabolism in alterations of cardiac function?

Rebs,

Streckfuss-Bömeke

2023

Front. Mol. Med

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Mutations in the mitochondrial-DNA or mitochondria related nuclear-encoded-DNA lead to various multisystemic disorders collectively termed mitochondrial diseases. One in three cases of mitochondrial disease affects the heart muscle, which is called mitochondrial cardiomyopathy (MCM) and is associated with hypertrophic, dilated, and noncompact cardiomyopathy. The heart is an organ with high energy demand, and mitochondria occupy 30%–40% of its cardiomyocyte-cell volume. Mitochondrial dysfunction leads to energy depletion and has detrimental effects on cardiac performance. However, disease development and progression in the context of mitochondrial and nuclear DNA mutations, remains incompletely understood. The system of induced pluripotent stem cell (iPSC)-derived cardiomyocytes (CM) is an excellent platform to study MCM since the unique genetic identity to their donors enables a robust recapitulation of the predicted phenotypes in a dish on a patient-specific level. Here, we focus on recent insights into MCM studied by patient-specific iPSC-CM and further discuss research gaps and advances in metabolic maturation of iPSC-CM, which is crucial for the study of mitochondrial dysfunction and to develop novel therapeutic strategies.

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Human induced pluripotent stem cells for studying mitochondrial diseases in the heart

Cited by 10 publications

References 95 publications

Isotopic Tracing of Nucleotide Sugar Metabolism in Human Pluripotent Stem Cells

Isotopic Tracing of Nucleotide Sugar Metabolism in Human Pluripotent Stem Cells

Small molecule-mediated rapid maturation of human induced pluripotent stem cell-derived cardiomyocytes

How can we use stem cell-derived cardiomyocytes to understand the involvement of energetic metabolism in alterations of cardiac function?

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