Cell surface oxygen consumption by mitochondrial gene knockout cells

Herst, Patries M.; Tan, Ah-Hwee; Scarlett, Debbie-Jane G; Berridge, Michael V.

doi:10.1016/j.bbabio.2004.01.008

Cited by 97 publications

(83 citation statements)

References 35 publications

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“…Only few studies have looked at ROS generation in cells with mtDNA mutations [28][29][30]. It is debatable whether mtDNA-depletion in rho°cells generates more ROS than control cells would [31][32][33][34]. In the present study, the A3243G mutation was associated with higher intracellular ROS levels than control cells when glucose was removed from the medium.…”

Section: Discussionsupporting

confidence: 47%

Diabetes-associated mitochondrial DNA mutation A3243G impairs cellular metabolic pathways necessary for beta cell function

Andrade¹,

Rubı́²,

Frigerio³

et al. 2006

Diabetologia

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Aims/hypothesis Mitochondrial DNA (mtDNA) mutations cause several diseases, including mitochondrial inherited diabetes and deafness (MIDD), typically associated with the mtDNA A3243G point mutation on tRNALeu gene. The common hypothesis to explain the link between the genotype and the phenotype is that the mutation might impair mitochondrial metabolism expressly required for beta cell functions. However, this assumption has not yet been tested. Methods We used clonal osteosarcoma cytosolic hybrid cells (namely cybrids) harbouring mitochondria derived from MIDD patients and containing either exclusively wild-type or mutated (A3243G) mtDNA. According to the importance of mitochondrial metabolism in beta cells, we studied the impact of the mutation on key parameters by comparing stimulation of these cybrids by the main insulin secretagogue glucose and the mitochondrial substrate pyruvate. Results Compared with control mtDNA from the same patient, the A3243G mutation markedly modified metabolic pathways leading to a high glycolytic rate (2.8-fold increase), increased lactate production (2.5-fold), and reduced glucose oxidation (−83%). We also observed impaired NADH responses (−56%), negligible mitochondrial membrane potential, and reduced, only transient ATP generation. Moreover, cybrid cells carrying patient-derived mutant mtDNA exhibited deranged cell calcium handling with increased cytosolic loads (1.4-fold higher), and elevated reactive oxygen species (2.6-fold increase) under glucose deprivation. Conclusions/interpretation The present study demonstrates that the mtDNA A3243G mutation impairs crucial metabolic events required for proper cell functions, such as coupling of glucose recognition to insulin secretion.

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Section: Discussionsupporting

confidence: 47%

Diabetes-associated mitochondrial DNA mutation A3243G impairs cellular metabolic pathways necessary for beta cell function

Andrade¹,

Rubı́²,

Frigerio³

et al. 2006

Diabetologia

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“…due to the seeding procedure one day before, could have caused a lowered absorbance (A450 nm -A630 nm) compared to cells seeded conventionally due to the fact that stress seems to reduce WST-1 reduction [19]. Alternatively different seeding conditions and/or growth conditions could change oxygen supply resulting in an altered amount of superoxide which is generated by oxygen reduction and which is responsible for WST-1 reduction [20,21,22]. Another reason might be that a major part of cells attached on the HA surface have died [23].…”

Section: Discussionmentioning

confidence: 99%

Biocompatibility of ceramic scaffolds for bone replacement made by 3D printing

Leukers¹,

Gülkan²,

Irsen

et al. 2005

Mat.-wiss. u. Werkstofftech.

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Bone replacement materials used in tissue engineering require a high degree of safety and biological compatibility. For these reasons synthetic bone replacement materials based on calcium-phosphates are being used more widely. To mimic natural bone, rapid prototyping processes and especially 3D printing are favourable. Using 3D printing, complex 3 dimensional structures can be made easily.In this study we successfully performed biocompatibility tests with a Hydroxyapatite test structure (HA-S) made by 3D printing. Cytotoxicity tests were carried out according to DIN ISO 10993-5 in static and dynamic cultivation setups. To estimate cell proliferation and analyze morphology, histological evaluation was done. In summary, good cell viability as well as good proliferation behaviour were found. Moreover, these results show that the 3D printing process in combination with the suitable material presented in this study is well suited for fabricating scaffolds for TE in the required accuracy and biological compatibility.Key words: Scaffold fabrication, cell culture, biocompatibility, hydroxyapatite, 3D printing Knochenersatzmaterialien für das Tissue Engineering (TE) erfordern ein hohes Maß an Sicherheit und biologischer Verträglichkeit. Aus diesen Gründen nehmen synthetische Knochenersatzmaterialien auf der Basis von Calciumphosphaten einen immer größer werdenden Stellenwert ein. Um der Struktur des natürlichen Knochens so nahe wie möglich zu kommen, bieten sich Verfahren aus dem Rapid Prototyping, insbesondere das 3D Drucken an. Dadurch lassen sich komplexe, dreidimensionale Strukturen für das TE einfach herstellen.In dieser Studie haben wir erfolgreich eine mittels 3D Drucken hergestellte Struktur (HA-S) auf der Basis von Hydroxylapatit auf ihre Biokompatibilität getestet. Zytotoxizitätsuntersuchungen wurden analog DIN ISO 10993-5 in statischer und dynamischer Kultivierung durchgeführt. Zudem wurden histologische Schnitte zur Beurteilung der Zellproliferation und -morphologie durchgeführt. Insgesamt konnte eine gute Zellvitalität und Zellproliferation nachgewiesen werden. Darüber hinaus zeigen die Ergebnisse, dass das hier vorgestellte 3D Druckverfahren mit dem hier verwendeten Material dazu geeignet ist, Strukturen für das TE in ausreichender Prä-zision und biologischer Verträglichkeit herzustellen.

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“…In contrast, glycolytic cells recycle NADH via lactate dehydrogenase and PMET. [18][19][20] Although cancer cells exhibit elevated PMET, several orders of magnitude higher than their non-proliferative counterparts, 18 different types of cancer cells rely on PMET for NADH recycling to different extents, depending on their level of glycolytic metabolism. 21 Nevertheless, PMET provides an excellent target for anti-cancer drug development, especially in the treatment of highly refractory glycolytic cancers.…”

Section: Introductionmentioning

confidence: 99%

The anti-cancer drug, phenoxodiol, kills primary myeloid and lymphoid leukemic blasts and rapidly proliferating T cells

Herst¹,

Davis²,

Neeson³

et al. 2009

Haematologica

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The online version of this article contains a supplementary appendix. BackgroundThe redox-active isoflavene anti-cancer drug, phenoxodiol, has previously been shown to inhibit plasma membrane electron transport and cell proliferation and promote apoptosis in a range of cancer cell lines and in anti-CD3/anti-CD28-activated murine splenocytes but not in non-transformed WI-38 cells and human umbilical vein endothelial cells. Design and MethodsWe determined the effects of phenoxodiol on plasma membrane electron transport, MTT responses and viability of activated and resting human T cells. In addition, we evaluated the effect of phenoxodiol on the viability of leukemic cell lines and primary myeloid and lymphoid leukemic blasts. ResultsWe demonstrated that phenoxodiol inhibited plasma membrane electron transport and cell proliferation (IC50 46 µM and 5.4 µM, respectively) and promoted apoptosis of rapidly proliferating human T cells but did not affect resting T cells. Phenoxodiol also induced apoptosis in T cells stimulated in HLA-mismatched allogeneic mixed lymphocyte reactions. Conversely, non-proliferating T cells in the mixed lymphocyte reaction remained viable and could be restimulated in a third party mixed lymphocyte reaction, in the absence of phenoxodiol. In addition, we demonstrated that leukemic blasts from patients with primary acute myeloid leukemia (n=22) and acute lymphocytic leukemia (n=8) were sensitive to phenoxodiol. The lymphocytic leukemic blasts were more sensitive than the myeloid leukemic blasts to 10 µM phenoxodiol exposure for 24h (viability of 23±4% and 64±5%, respectively, p=0.0002). ConclusionsThe ability of phenoxodiol to kill rapidly proliferating lymphocytes makes this drug a promising candidate for the treatment of pathologically-activated lymphocytes such as those in acute lymphoid leukemia, or diseases driven by T-cell proliferation such as autoimmune diseases and graft-versus-host disease.

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Cell surface oxygen consumption by mitochondrial gene knockout cells

Cited by 97 publications

References 35 publications

Diabetes-associated mitochondrial DNA mutation A3243G impairs cellular metabolic pathways necessary for beta cell function

Diabetes-associated mitochondrial DNA mutation A3243G impairs cellular metabolic pathways necessary for beta cell function

Biocompatibility of ceramic scaffolds for bone replacement made by 3D printing

The anti-cancer drug, phenoxodiol, kills primary myeloid and lymphoid leukemic blasts and rapidly proliferating T cells

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