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Posts Tagged ‘Hallmarks of Cancer’

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Hypoxia and the Hallmarks of Cancer: Metabolic Reprogramming

Hanahan and Weinberg’s seminal papers on the Hallmarks of Cancer describe how cancer cells accommodate the frenzied growth characteristic of tumours. Low oxygen is eminently characteristic of tumours, and in this hypoxic environment, metabolism is reprogrammed to satisfy energetic and synthetic needs of the cells.

 

Our series on Hypoxia and the Hallmarks of Cancer has showcased research on how hypoxia in the tumour microenvironment affects 8 of the Hallmarks, and in the fifth and final chapter, we look more closely at how researchers are using the Hypoxystation to delineate the Hallmark Metabolic Reprogramming.

The Hypoxystation creates authentic cell culture conditions with regard to oxygen, CO2, temperature, and humidity.  Glove-less access to culture and manipulate cells under physiological atmosphere, in a HEPA-clean environment, allows cancer researchers to re-create the hypoxic tumour microenvironment. Hypoxystation user Dr Ali Tavassolli states that “We have only ever used the H35. I like the ease with which we can regulate and change the oxygen concentration”. And our user Dr. Brad Wouters at the Princess Margaret Cancer Centre in Toronto, who recently purchased his fourth Hypoxystation, says, “The continuous hypoxia we achieve in the workstation is a prerequisite for studies with hypoxia-activated drugs used in cancer therapy strategies.”

Hallmarks of Cancer

Metabolic Reprogramming

Changes in energy metabolism feature prominently in aggressive malignancy, and tumour hypoxia and the responding signalling pathways, featuring many HIF target genes, clearly interface with reprogrammed tumour metabolism. Reprogramming of conventional metabolic pathways serves to satisfy burgeoning energetic and anabolic needs of the tumour cells; many cancer cells may preferentially utilise glycolysis over oxidative phosphorylation, uncoupling mitochondrial metabolism from oxygen availability. Hypoxia-induced HIF’s attenuate mitochondrial function through diverse mechanisms, including down-regulation of enzymes in the electron transport chain and suppression of biogenesis of mitochondria. Signalling pathways involving HIF’s and many products of oncogenes and tumour suppressor genes interact to balance the energy needs of dividing cells with the requirement for bio-synthetic intermediates. Activation of lipid biosynthesis and other pathways with biosynthetic significance, such as the pentose phosphate pathway, is another metabolic consequence of hypoxia and HIF up-regulation. Reactive oxygen species ROS produced by the mitochondria stabilise HIF-1, influence redox homeostasis, and provide protective antioxidants to the cancer cells.

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Clostridium difficile studies can be done in a Whitley Workstation

Hallmarks of Cancer: Sustaining Growth and Resisting Cell Death

In part four of our mini-series describing “Hypoxia and the Hallmarks of Cancer”, we look more closely at how researchers are using the Hypoxystation to delineate the Hallmarks Sustaining Growth and Resisting Cell Death.

 

 

 

 

 

Hallmarks of Cancer

Resisting Cell Death

The ability of cells to resist cell death under hypoxic conditions is central to the progression of cancer and the acquisition of resistance to chemotherapy so frequently encountered in tumors. Hypoxia in the tumor microenvironment exerts selective pressure favoring cells that have lost the functionality of apoptosis genes and can expand uncontrollably.  Hypoxia also contributes to survival by inducing autophagy, in a pathway involving HIF-1, beclin, BNIP3 and BNIP3L, in which cellular autophagy acts to recycle cellular organelles, satisfy metabolic demand and improve hypoxic tolerance.  HIF-1 mediates cell-cycle retardation and arrest, causing hypoxic tumor cells to become resistant to radiotherapies. NF-κB, through its effects on myriad transcription factors, for example through inhibition of cell death signalling, is activated by hypoxia and reactive oxygen species, and also promotes cell survival.

Sustaining Growth

Cancer is essentially based on the cells’ inability to “stop” when suppressors signal an end to growth, and the compunction to “go” despite a lack of bonafide growth signals. Hypoxia in the context of cancer, in precipitating genomic instability and mutation, results in numerous inactive tumor suppressor genes and activated growth factor genes, such that the combination of constitutive proliferative signaling and mutated cancer genes leads to sustained growth. HIF and NF-κB regulated pathways involving Notch, mTOR, WNT11, CAIX, and IGF-1, among many others, contribute to sustained growth in cancer as regulation of proliferation derails. Induced by hypoxia-regulated proteins, anabolic pathways for nucleotide and lipid synthesis are ramped up and enable the rapid proliferation typical of cancer.

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Hypoxia and the Hallmarks of Cancer: Angiogenesis and Metastasis

The following was provided by HypOxygen, our distributor of Hypoxic Workstations in the US – Hanahan and Weinberg’s “Hallmarks of Cancer” are at the root of the multi-step progression of cancer, and they are all influenced by hypoxia in the tumor microenvironment. In this mini-review series, HypOxygen has been taking a closer look at the way Hypoxystation users worldwide are delineating the effects of hypoxia on the Hallmarks of Cancer: so far, we’ve showcased Avoiding Immune Destruction and Tumour Promoting Inflammation and Genome Instability and Mutation and Enabling Replicative Immortality.

In the Hypoxystation, researchers working with cells in culture can mimic the physiological conditions that produce those characteristic Hallmarks. The Hypoxystation enables glove-less access to cultivate and manipulate cells under physiological conditions, in a HEPA-clean environment. Oxygen levels in the Hypoxystation can be reliably and accurately adjusted to below 1%, reflecting the high metabolism, low perfusion tumor microenvironment.

 

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Hallmarks Of Cancer
1. Inducing Angiogenesis

Angiogenesis and tumor-associated neo-vascularization are central to the progression of cancer, and hypoxia in the fast-growing, poorly perfused tumor setting is one of the main factors driving the formation of new vessels. Hypoxia in the tumor activates the hypoxia stress response, which is mediated at the cellular level by HIF, VEGF and many other cytokines, growth factors and guidance molecules. As a consequence, endothelial cells and pericytes proliferate and form new blood vessels, which are, however, disorderly and leaky, in turn exacerbating hypoxia in the tumor. Cancer treatment strategies striving to normalize tumor vessels for the purpose of improved drug delivery and alleviation of hypoxia in the tumor are showing great promise.

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2. Activating Invasion and Metastasis

As with the other Hallmarks of Cancer, metastasis and cancer progression are correlated with low oxygen levels in the tumor. HIF’s activate the expression of more than 1000 genes, numerous of which play a role in inducing genes involved in the EMT, through direct interactions with HRE’s at promotor sites and other mechanisms such as epigenetic alterations, like methylation/demethylation. Hypoxia promotes migration and invasion by facilitating the endothelial-mesenchymal transition, altering cell-cell contacts, and reducing adhesion to the extra-cellular matrix. Cancer cells and neighboring cells such as fibroblasts are all influenced by hypoxia, and all contribute to the restructuring of the tumor microenvironment. The effects of the Hallmarks of Cancer continually perturb and promote each other, as when hypoxia-driven metabolic reprogramming causes acidification of the extracellular microenvironment through increased production and secretion of lactate, in turn augmenting ECM remodeling and immune evasion. Similarly, formation of novel blood vessels enables extravasation and migration of cancer cells to form new tumors.

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The Hallmarks of Cancer: Genome Instability and Immortality

 

Dr Burga Kalz Fuller continues to look at the way the iconic “Hallmarks of Cancer“, as first described by Douglas Hanahan and Robert Weinberg, are influenced by hypoxia in the tumour microenvironment.

Oxygen around and within the tumour cells is central to metabolism, immunology, epigenetics and therapy resistance of all the cancers; in the lab, oxygen levels during tumour cell culture exert effects on metabolism, maintenance, cell yield, and cell survival. That’s why the authentic physiological cell culture conditions in the Hypoxystation help advance research into tumour progression and other events which determine malignancy and outcome of cancer diseases. The Hypoxystation enables glove-less access to cultivate and manipulate cells under physiological conditions, in a HEPA-clean environment.

In this mini-review series, we take a closer look at the way Hypoxystation users worldwide are delineating Hypoxia and the Hallmarks of Cancer. Previously, we had showcased research by Hypoxystation users involved with Avoiding Immune Destruction and Tumour Promoting Inflammation. Next, we want to show the many ways in which Hypoxystation users are researching the Hallmarks Genome Instability and Mutation and Enabling Replicative Immortality. One of those researchers, Dr. David Ho of the University of Miami, presented his results at the Cell Symposium on Cancer, Inflammation and Immunity in San Diego in June.


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David Ho

Dr. David Ho from the University of Miami with his poster presentation at the Cell Symposium on Cancer, Inflammation and Immunity

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1. Genome Instability and Mutation

Tumour hypoxia drives genomic instability both by increasing the volume of mutations (DNA strand breaks, base damage, and gene amplification) and by diminishing DNA repair efficiency. The low levels of oxygen typical of the tumor microenvironment decrease transcription of genes related to homologous repair and non-homologous end-joining, leading to the genetic instability observed in hypoxic tumour cells. Hypoxia induces production of reactive oxygen species ROS, which interact with nucleic acids, proteins and lipids, causing cellular damage and mutagenesis. Hypoxic activation of HIF-1 also upregulates expression of certain miRNA’s which suppress DNA repair pathways.

 

LITERATURE:

  • Jiang et al. (2016) “Hypoxia Potentiates the Radiation-Sensitizing Effect of Olaparib in Human Non-Small Cell Lung Cancer Xenografts by Contextual Synthetic Lethality” Int J Radiation Oncol Biol Phys, Vol. 95, No. 2, pp. 772 e781, 2016
    www.redjournal.org/article/S0360-3016(16)00056-0/abstract Hypoxystation user
  • Doherty et al. (2016) “Photodynamic killing of cancer cells by a Platinum(II) complex with cyclometallating ligand” Nature Scientific Reports 6:22668 (2016)
    www.ncbi.nlm.nih.gov/pmc/articles/PMC4778139/ Hypoxystation user
  • Hunter et al. (2016) “Hypoxia-activated prodrugs: paths forward in the era of personalised medicine” Br J Cancer. 2016 May 10; 114(10): 1071–1077
    www.ncbi.nlm.nih.gov/pmc/articles/PMC4865974/Hypoxystation user
  • Leszczynska et al. (2016) “Mechanisms and consequences of ATMIN repression in hypoxic conditions: roles for p53 and HIF-1” Scientific Reports 6:21698 (2016
    www.ncbi.nlm.nih.gov/pmc/articles/PMC4753685/ Hypoxystation user
  • Timpano and Uniacke (2016) “Human Cells Cultured Under Physiological Oxygen Utilize Two Cap-binding Proteins to Recruit Distinct mRNAs for Translation” Journal of Biological Chemistry 291(20):jbc.M116.717363
    www.jbc.org/content/291/20/10772.abstract Hypoxystation user
  • Haider et al. (2016) “Genomic alterations underlie a pan-cancer metabolic shift associated with tumour hypoxia“ Genome Biology (2016) 17:140
    www.ncbi.nlm.nih.gov/pubmed/27358048

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2. Enabling Replicative Immortality

Cancer is characterized by a nearly unlimited capacity of the tumour cells to proliferate. Hypoxia in the rapidly growing tumour supports immortalisation of a subset of cancer cells, the “cancer stem cells”. Factors such as hypoxia in the tumour microenvironment derail signals indicating senescence and initiating apoptosis, enabling an immortal lifespan. Telomerase, Notch, c-Myc, and OCT4 mediate the acquisition of a stem cell-like phenotype through down-regulation of differentiation genes and activation of stem genes, generating CSC’s with aggressive properties. These cancer stem cells residing in an hypoxic tumour niche are uniquely resistant to many therapies, where low oxygen promotes stemness, maintenance, and self-renewal of the CSC’s. Metastasis and invasion by these CSC’s induce the formation of secondary tumours, which in most cases dramatically worsen the prognosis for cancer patients.

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Scientist Working in Whitley Workstation

Hypoxia in the Tumour Microenvironment

Hypoxia in the tumour microenvironment affects all the characteristic Hallmarks of Cancer, significantly impacting progression of the cancer and the patients’ prognosis. Inflammation and immunity are both acutely influenced by the low oxygen typical of the tumour microenvironment: hypoxia creates an immune-suppressive network supporting tumour growth and metastasis, and it induces sustained inflammation in a “wound that never heals”.

Cancer research depends on recreating a physiologically accurate environment for cell cultures in the lab, and hypoxia in a closed workstation format such as a Whitley Hypoxystation is the best way to do that. Incubate, image, manipulate and assay – all inside the continuous, reliably stable hypoxic environment. HEPA filtered air scrubbed to ISO 14644 class 3 standards, sterile humidity, and containment options make the Hypoxystation the safest, cleanest workstation available for hypoxic cell culture down to 0.1% O2.

Our Hypoxystation users are investigating all aspects of the Hallmarks of Cancer and how they are shaped by hypoxia. We review their recent research on Avoiding Immune Destruction and Tumour Promoting Inflammation here.

 

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HypOxygen at Tumour Microenvironment Workshop in Miami

This article was written by Burga Kalz Fuller, join her and HypOxygen at the 15th International Tumour Microenvironment Workshop in Miami

In most cancers, the hypoxic microenvironment affects the development and progression of tumours, driving alterations in gene expression, metabolism and cell signalling, and significantly influencing the Hallmarks of Cancer. So what about in vitro cancer research, do culture parameters matter? Definitely! Numerous studies have shown that even very brief exposure to ambient oxygen levels and temperature significantly impacts cell culture, behaviour and function of cells in vitro.

HypOxygen will be exhibiting our Hypoxystation at the 15th International Tumour Microenvironment Workshop in Miami from 27th – 29th April. The special focus there is on “Hypoxia, Angiogenesis and Vasculature”, reflecting the critical importance of hypoxia in the context of cancer. With the Hypoxystation, cancer researchers have their finger on the pulse of physiological cell culture.

 

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The Hypoxystation mimics the hypoxic conditions present in cancer, providing a closed workstation format with contiguous, stable low oxygen down to 0.1%. Precise oxygen, carbon dioxide, and humidity control within a temperature-controlled environment, as well as ample space for cellular manipulation, assays and microscopic observation, allow researchers to recreate physiological conditions. HEPA filtration, sterile steam humidification, and remote parameter monitoring are some of the features that make the Hypoxystation so unique.

Cancer research labs, who use a Hypoxystation to re-create hypoxic conditions in the tumour microenvironment, are publishing brilliant papers which demonstrate the influence of hypoxia on the Hallmarks of Cancer. Metabolic adaptation, sustained growth, resisting cell death, and angiogenesis are just some of the Hallmarks which are affected by hypoxia. Here are some recent highlights:

Hypoxystation users are showing that “culturing cells in ambient air, or ‘normoxia’ is far from physiological.

Visit HypOxygen at the 15th International Tumor Microenvironment Workshop in Miami

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Hypoxia and the Hallmarks of Cancer

Therapeutic Targeting of Hypoxia and HIFs in Cancer. Dr Burga Kalz Fuller from US distributor HypOxygen has summarised this study that outlines the Hallmarks of Cancer.

“Tumour hypoxia and HIFs affect most of the cancer hallmarks… and contribute to chemo- and radiotherapy resistance.” In their review from 2016, Wigerup, Pahlman and Bexell of Lund University in Sweden discuss how hypoxia inducible factors HIFs regulate the hypoxic microenvironment in cancer, and the therapeutic strategies that are being developed to improve patients’ prognosis. Dr. Sven Pahlman’s lab has been using the H35 Hypoxystation for more than 5 years, to research SCLC and neuroblastoma, and their data is contributing to the understanding of the role of oxygen levels in the progression of cancer.

Hypoxia and HIF-1α and 2α expression in cancer usually signify a worse prognosis, but most hypoxia-induced transcriptional, translational, and epigenetic changes are cell-type specific. Many effects engendered by hypoxia are mediated directly or indirectly via HIF pathways, and most are causative of the iconic “Hallmarks of Cancer” that Hanahan and Weinberg introduced in 2000 and expanded in 2011. Hypoxia induces increased autophagy, apoptosis, and aberrant cell proliferation; neoangiogenesis mediated by VEGF and PDGF-β; proliferation of cancer stem cells; metabolic reprogramming to satisfy energy and synthetic requirements in proliferating cells; modulation of inflammation and immune responses; genomic instability through increased mutagenesis and diminished DNA repair; and metastasis as hypoxia induces epithelial-to-mesenchymal transition and degradation of the extracellular matrix. Assaying the relationship between hypoxia and the Hallmarks of Cancer benefits significantly from the physiological atmosphere mimicked in the Hypoxystation, a closed-culture hypoxia workstation controlling gasses, temperature and humidity.


Visit Don Whitley Scientific and HypOxygen at

Keystone Adaptations to Hypoxia and Tumour Metabolism

Location: Whistler, BC  Date: 5th – 9th March

Sven Pahlman


In their review, Wigerup and Pahlman describe the role tumour hypoxia plays for cancer therapy and treatment resistance, as oxygen levels, production of reactive oxygen species ROS, and HIF activity are intertwined actors in the cancer battle. Any and all effects of hypoxia are cell-type specific; however, numerous studies indicate that HIF’s mediate chemoresistance, suggesting that HIF-1 and 2 inhibitors can effectively support cancer therapy. The authors state that “since hypoxia is a hallmark of solid tumours and mediates aggressive, metastatic, and resistant disease, it is arguably one of the most attractive therapeutic targets in cancer.” Strategies selectively targeting hypoxia for cancer therapy include hypoxia-activated prodrugs; inhibitors of HIF mRNA and protein expression; and inhibitors of downstream HIF signalling pathways such as VEGF. Effective drug research relies on authentic replication of the hypoxic environment for cell culture: the Hypoxystation used in the Pahlman lab is able to accommodate long-term assays with sterile steam humidification and HEPA clean air. The Hypoxystation concept “Choose your Atmosphere – Define your Environment” is the best way to ensure cell culture reflects physiology in cancer research and therapy.

Hypoxia is at the heart of the Hallmarks of Cancer, and results such as these from the Pahlman lab make the cancer research community hopeful that “HIF inhibition is likely to be a powerful therapeutic approach” to eradicate cancer.

 

Hallmarks of Cancer

KONICA MINOLTA DIGITAL CAMERA

The Hallmarks of Cancer

The Hallmarks of Cancer are a specific set of characteristics that are inherent to cancer. The Hallmarks were published by Hanahan and Weinberg in 2000 (updated in 2011) and have become extremely recognisable in the cancer research community both as a scientific concept and as a strong, visual image.

The Hallmarks of Cancer have been an area of study for several years and a key focus of research into causes and progression of cancer. One such study by a lab in Sweden using the H35 Hypoxystation, entitled “Therapeutic targeting of hypoxia and hypoxia-inducible factors in cancer” by Wigerup, Pahlman and Bexell links cancer characteristics with hypoxia as an underlying cause.  This review of hypoxia-driven cancer characteristics and tumour progression makes a crucial connection between hypoxia and the “Hallmarks of Cancer”, a set of specific characteristics that are inherent to cancer. There are many more publications showing that hypoxia is intimately involved in every aspect of the disease complex cancer.

The image below summarises the 9 Hallmarks of Cancer. The Hypoxystation in the middle of the graphic symbolises how the low oxygen environment re-creates the atmosphere where cancer cells are required to act in a physiological manner. The dial around the Hypoxystation indicates the different levels of oxygen required for specific types of cancer work. Ultimately, the graphic shows how the Hypoxystation facilitates a level of oxygen that cannot be achieved reliably in an incubator, and which is necessary to effectively research cancer therapies.

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Graphic provided by HypOxygen

 

 

 

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Hallmarks of Cancer event in Belgium

Over the last few days Don Whitley Scientific has been exhibiting at the Cell Symposia Hallmarks of Cancer event in Ghent, Belgium. This was an event that gathered global leaders in cancer research to enhance understanding of the key aspects of cancer.  

Delegates at the event were able to visit the Don Whitley Scientific exhibition stand to find out more about how hypoxic workstations are relevant for cancer research applications. Visitors to the stand could interact with a Whitley i2 Instrument Workstation , a workstation with a large capacity developed in response to a rising number of enquiries from scientists who wanted to use Seahorse Extracellular Flux (XF) Analyzers in hypoxic conditions.

 

There is a wealth of data showing that hypoxia in tumours is driving the progression of cancer and that important characteristics of cancer are influenced, or even caused, by a pathological lack of oxygen.

 

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Don Whitley Scientific at Cell Symposia Hallmarks of Cancer, Ghent, 2018

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