Modeling the dynamics of hypoxia inducible factor-1α (HIF-1α) within single cells and 3D cell culture systems

Leedale, Joseph; Herrmann, Anne‐Kristin; Bagnall, James; Fercher, Andreas; Papkovsky, Dmitri B.; Sée, Violaine; Bearon, Rachel N.

doi:10.1016/j.mbs.2014.09.007

Cited by 25 publications

(15 citation statements)

References 33 publications

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“…Computational modeling of the temporal dynamics of the HIF-1α response to hypoxia has indicated that regulation of HIF-1α is can be clearly mapped to the presence of defined negative feedback loops ( Cavadas et al, 2013 ). These include the activation of HIF-1α regulated prolyl hydroxylase (PHD) 2 and 3 enzymes ( Leedale et al, 2014 ), as well as other HIF regulators such as miR155 ( Bruning et al, 2011 ), miR-199a-5p, miR-93, and cRel ( Jiang et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Glycolytic Response to Inflammation Over Time: Role of Myeloid HIF-1alpha

et al. 2018

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The in vivo response to lipopolysaccharide (LPS) occurs rapidly and has profound physiological and metabolic effects. The hypoxia inducible (HIF) transcription factor is an intrinsic and essential part of inflammation, and is induced by LPS. To determine the importance of the HIF response in regulating metabolism following an LPS response, glucose uptake was quantified in a time dependent manner in mice lacking HIF-1α in myeloid cells. We found that deletion of HIF-1α has an acute protective effect on LPS-induced hypoglycemia. Furthermore, reduced glucose uptake was observed in the heart and brown fat, in a time dependent manner, following loss of HIF-1α. To determine the physiological significance of these findings, cardiovascular, body temperature, and blood pressure changes were subsequently quantified in real time using radiotelemetry measurements. These studies reveal the temporal aspects of HIF-1α as a regulator of the metabolic response to acute LPS-induced inflammation.

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Section: Introductionmentioning

confidence: 99%

Glycolytic Response to Inflammation Over Time: Role of Myeloid HIF-1alpha

et al. 2018

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“…may cause the exact value to vary. However, it should be noted that the values used in the model appear feasible for the system and are the same order of magnitude as values found in other multicellular spheroid studies (Leedale et al, 2014, Lesher-Pérez et al, 2017. The Michaelis constant appears to be more difficult to ascertain as only measurements based upon rat studies have been made.…”

Section: Model Parameterisationmentioning

confidence: 77%

“…To determine oxygen transport parameters, the rate of oxygen diffusion through the spheroid was derived based on the modelling results of Leedale et al (2014). This previous model assumed linear oxygen consumption kinetics and optimised parameter values by fitting to oxygen distributions in cellular spheroids, measured under various external oxygen conditions.…”

Section: Model Parameterisationmentioning

confidence: 99%

“…The importance of understanding the local environment and mechanisms within tumour spheroids to develop medical applications has led to considerable mathematical modelling efforts in this area. Some work has focused on reproducing growth and development processes (Chaplain, 1996, while others investigate the effects of various oxygen consumption kinetics in different geometries and hypoxic environments (Grimes et al, 2014b, Grimes et al, 2014a, Leedale et al, 2014. Acknowledgement of the importance of spatial dynamics in spheroids has also led to mathematical models that simulate 3D pharmacological processes such as drug delivery and metabolism (Ward & King, 2003, Mehta et al, 2012.…”

Section: Introductionmentioning

confidence: 99%

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In silico-guided optimisation of oxygen gradients in hepatic spheroids

Leedale

Colley

Gaskell

et al. 2019

Computational Toxicology

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One of the key advantages of assessing the hepatotoxic potential of xenobiotics in spheroids rather than monolayer cell culture is the existence of a more physiologically relevant testing environment. Three-dimensional cultures support spatial gradients in nutrients such as oxygen that can be exploited to better represent in vivo gradients that exist along a fundamental subunit of liver microarchitecture, the liver sinusoid. The physical and physiological processes that result in the establishment of such gradients can be described mathematically. Quantification of the rates governing these processes and optimisation of cell culture conditions can be performed in silico to better inform experimental design. In this study, we take into account cell line-specific physiological properties, spheroid size and the impact of experimental equipment geometries in order to demonstrate how mathematical models can be optimised to achieve specific in vivo-like features in different scenarios. Furthermore, the sensitivity of such optimised gradients is analysed with respect to culture conditions and considerations are given to prevent the emergence of hypoxic regions in the spheroid. The methodology presented provides an enhanced understanding of the mechanisms of the system within this simulated in vitro framework such that experimental design can be more carefully calibrated when conducting experiments using hepatic spheroids.

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“…This approach, exemplified in Fig. 4 by the map of theoretically predicted subcellular PHD2 activity, may be particularly attractive for systems biology and hypoxia signaling research (34,45). At this stage, modeling of local PHD activity is mostly hypothetical, because our iO 2 maps are generated ex vivo.…”

Section: Discussionmentioning

confidence: 99%

Imaging of oxygen gradients in giant umbrella cells: an ex vivo PLIM study

Zhdanov

Golubeva

Okkelman

et al. 2015

American Journal of Physiology-Cell Physiology

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O2 plays a pivotal role in aerobic metabolism and regulation of cell and tissue function. Local differences and fluctuations in tissue O2 levels are well documented; however, the physiological significance of O2 microgradients, particularly at the subcellular level, remains poorly understood. Using the cell-penetrating phosphorescent O2 probe Pt-Glc and confocal fluorescence microscopy, we visualized O2 distribution in individual giant (>100-μm) umbrella cells located superficially in the urinary bladder epithelium. We optimized conditions for in vivo phosphorescent staining of the inner surface of the mouse bladder and subsequent ex vivo analysis of excised live tissue. Imaging experiments revealed significant (≤85 μM) and heterogeneous deoxygenation within respiring umbrella cells, with radial O2 gradients of up to 40 μM across the cell, or ∼0.6 μM/μm. Deeply deoxygenated (5-15 μM O2) regions were seen to correspond to the areas enriched with polarized mitochondria. Pharmacological activation of mitochondrial respiration decreased oxygenation and O2 gradients in umbrella cells, while inhibition with antimycin A dissipated the gradients and caused gradual reoxygenation of the tissue to ambient levels. Detailed three-dimensional maps of O2 distribution potentially can be used for the modeling of intracellular O2-dependent enzymatic reactions and downstream processes, such as hypoxia-inducible factor signaling. Further ex vivo and in vivo studies on intracellular and tissue O2 gradients using confocal imaging can shed light on the molecular mechanisms regulating O2-dependent (patho)physiological processes in the bladder and other tissues.

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Modeling the dynamics of hypoxia inducible factor-1α (HIF-1α) within single cells and 3D cell culture systems

Cited by 25 publications

References 33 publications

Glycolytic Response to Inflammation Over Time: Role of Myeloid HIF-1alpha

Glycolytic Response to Inflammation Over Time: Role of Myeloid HIF-1alpha

In silico-guided optimisation of oxygen gradients in hepatic spheroids

Imaging of oxygen gradients in giant umbrella cells: an ex vivo PLIM study

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