Mitophagy mediates metabolic reprogramming of induced pluripotent stem cells undergoing endothelial differentiation

Krantz, Sarah; Kim, Young-Mee; Srivastava, Shubhi; Leasure, Joseph W.; Tóth, Péter T.; Marsboom, Glenn; Rehman, Jalees

doi:10.1016/j.jbc.2021.101410

Cited by 17 publications

(22 citation statements)

References 39 publications

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“…In particular, shear stress should be lower than 0.2 Pa for hepatocytes to avoid cell sufferance, while the oxygen concentration should be higher than 0.021 mol/m 3 . Moreover, the results showed that the shear stress (ranged 0.1–1 Pa) experienced by endothelial cells on the membrane of the lower chamber were slightly lower than the in vivo values for capillaries (Figure e) …”

Section: Resultsmentioning

confidence: 87%

“…18 Moreover, the results showed that the shear stress (ranged 0.1−1 Pa) experienced by endothelial cells on the membrane of the lower chamber were slightly lower than the in vivo values for capillaries (Figure 4e). 19 Dynamic Culturing of the 3D Liver Model inside the Millifluidic Device. Based on the above-described computa- tional results, we passed on to dynamic culturing of our LoC.…”

Section: ■ Statisticsmentioning

confidence: 99%

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Development of an Induced Pluripotent Stem Cell-Based Liver-on-a-Chip Assessed with an Alzheimer’s Disease Drug

Fanizza

Boeri

Donnaloja

et al. 2023

ACS Biomater. Sci. Eng.

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Liver-related drug metabolism is a key aspect of pharmacokinetics and possible toxicity. From this perspective, the availability of advanced in vitro models for drug testing is still an open need, also to the end of reducing the burden of in vivo experiments. In this scenario, organ-on-a-chip is gaining attention as it couples a state-of-the art in vitro approach to the recapitulation of key in vivo physiological features such as fluidodynamics and a tri-dimensional cytoarchitecture. We implemented a novel liver-on-a-chip (LoC) device based on an innovative dynamic device (MINERVA 2.0) where functional hepatocytes (iHep) have been encapsulated into a 3D hydrogel matrix interfaced through a porous membrane with endothelial cells (iEndo)]. Both lines were derived from human-induced pluripotent stem cells (iPSCs), and the LoC was functionally assessed with donepezil, a drug approved for Alzheimer’s disease therapy. The presence of iEndo and a 3D microenvironment enhanced the expression of liver-specific physiologic functions as in iHep, after 7 day perfusion, we noticed an increase of albumin, urea production, and cytochrome CYP3A4 expression compared to the iHep static culture. In particular, for donepezil kinetics, a computational fluid dynamic study conducted to assess the amount of donepezil diffused into the LoC indicated that the molecule should be able to pass through the iEndo and reach the target iHep construct. Then, we performed experiments of donepezil kinetics that confirmed the numerical simulations. Overall, our iPSC-based LoC reproduced the in vivo physiological microenvironment of the liver and was suitable for potential hepatotoxic screening studies.

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Section: Resultsmentioning

confidence: 87%

Section: ■ Statisticsmentioning

confidence: 99%

Development of an Induced Pluripotent Stem Cell-Based Liver-on-a-Chip Assessed with an Alzheimer’s Disease Drug

Fanizza

Boeri

Donnaloja

et al. 2023

ACS Biomater. Sci. Eng.

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“…Human ovarian tumors show a high overrepresentation of mitophagy pathways ( 56 ). A recent report concluded that mitophagy is required for the adaptive reprogramming of cellular metabolism ( 57 ), and provides molecules for cell survival and enhanced stemness and stem cell renewal (see review ( 58 ). Further, BNIP3 has been shown to support migration via maintaining actin cytoskeleton plasticity and focal adhesion dynamics ( 59 ), linking hypoxia-induced mitophagy with cellular functions.…”

Section: Discussionmentioning

confidence: 99%

Mitochondrial plasticity supports proliferative outgrowth and invasion of ovarian cancer spheroids during adhesion

et al. 2023

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BackgroundOvarian cancer cells aggregate during or after exfoliation from the primary tumor to form threedimensional spheroids. Spheroid formation provides a survival advantage during peritoneal dissemination in nutrient and oxygen-depleted conditions which is accompanied by a suppressed metabolic phenotype and fragmented mitochondria. Upon arrival to their metastatic sites, spheroids adhere to peritoneal organs and transition to a more epithelial phenotype to support outgrowth and invasion. In this study, we investigated the plasticity of mitochondrial morphology, dynamics, and function upon adhesion.MethodsUsing our slow-developing (MOSE-L) and fast-developing (MOSE-LTICv) ovarian cancer models, we mimicked adhesion and reoxygenation conditions by plating the spheroids onto tissue culture dishes and changing culture conditions from hypoxia and low glucose to normoxia with high glucose levels after adhesion. We used Western Blot, microscopy and Seahorse analyses to determine the plasticity of mitochondrial morphology and functions upon adhesion, and the impact on proliferation and invasion capacities.ResultsIndependent of culture conditions, all spheroids adhered to and began to grow onto the culture plates. While the bulk of the spheroid was unresponsive, the mitochondrial morphology in the outgrowing cells was indistinguishable from cells growing in monolayers, indicating that mitochondrial fragmentation in spheroids was indeed reversible. This was accompanied by an increase in regulators of mitobiogenesis, PGC1a, mitochondrial mass, and respiration. Reoxygenation increased migration and invasion in both cell types but only the MOSE-L responded with increased proliferation to reoxygenation. The highly aggressive phenotype of the MOSE-LTICv was characterized by a relative independence of oxygen and the preservation of higher levels of proliferation, migration and invasion even in limiting culture conditions but a higher reliance on mitophagy. Further, the outgrowth in these aggressive cells relies mostly on proliferation while the MOSE-L cells both utilize proliferation and migration to achieve outgrowth. Suppression of proliferation with cycloheximide impeded aggregation, reduced outgrowth and invasion via repression of MMP2 expression and the flattening of the spheroids.DiscussionOur studies indicate that the fragmentation of the mitochondria is reversible upon adhesion. The identification of regulatory signaling molecules and pathways of these key phenotypic alterations that occur during primary adhesion and invasion is critical for the identification of druggable targets for therapeutic intervention to prevent aggressive metastatic disease.

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“…However, studies have found that the inhibition of mitophagy in cells could also increase mitochondrial mass 53 . This is because in somatic cells, mitophagy selectively eliminates damaged mitochondria, whereas in stem cells, mitophagy may serve as a physiologic mechanism for the metabolic rewiring of differentiating stem cells by removing old mitochondria and replacing them with new mitochondria configured for the differentiated cell state 49,52,54 . As in our study, cells with both higher mitochondrial mass and K14 (marker of proliferation) expression were localised in periphery of the colonies, indicating higher mitochondrial mass cells are proliferated cells, we suspect that the increased mitochondrial mass after short-term ROCKi treatment may be related to rapid differentiating KSCs to TA cells.…”

Section: Discussionmentioning

confidence: 99%

Short-term Rho-associated kinase inhibitor treatment accelerates primary keratinocyte growth while preserving stem cell characteristics

Jayarajan¹,

Hall

Xenakis

et al. 2022

Preprint

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Somatic stem cells can be cultured in-vitro and are attractive for cell and gene therapies, but their slow growth in in-vitro culture affects survival and stemness and hinders clinical applications. Rho-associated kinase inhibitor (ROCKi) has been used to overcome these obstacles. However, it risks changing the characteristics of stem cells. We found that primary keratinocyte stem cells (KSCs) cultured with the ROCKi Y-27632 for six days exhibited rapid proliferation while maintaining the ability to differentiate. Importantly, after discontinuation of ROCKi treatment, KSC numbers and characteristics were indistinguishable from those in non-treated cultures. We further confirmed that ROCKi treatment resulted in the activation of AKT and ERK pathways, which could support cell survival and proliferation in keratinocytes. We thus concluded that accelerating keratinocyte expansion with short-term ROCKi treatment does not exhaust KSCs' self-renewal and differentiation capacities, presenting a safe avenue for clinical applications.

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Mitophagy mediates metabolic reprogramming of induced pluripotent stem cells undergoing endothelial differentiation

Cited by 17 publications

References 39 publications

Development of an Induced Pluripotent Stem Cell-Based Liver-on-a-Chip Assessed with an Alzheimer’s Disease Drug

Development of an Induced Pluripotent Stem Cell-Based Liver-on-a-Chip Assessed with an Alzheimer’s Disease Drug

Mitochondrial plasticity supports proliferative outgrowth and invasion of ovarian cancer spheroids during adhesion

Short-term Rho-associated kinase inhibitor treatment accelerates primary keratinocyte growth while preserving stem cell characteristics

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