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University of Manchester
1.
Blount, Kathryn.
Cancer Systems Biology: Is the devil in the glycolytic
detail?.
Degree: 2014, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:222501
► An approach to investigating cancer that has recently seen resurgence of interest is the “Warburg effect”. Otto Warburg originally described the altered metabolism of cancer…
(more)
▼ An approach to investigating cancer that has
recently seen resurgence of interest is the “Warburg effect”. Otto
Warburg originally described the altered metabolism of cancer cells
and identified that they exhibit an increase in glucose uptake and
lactate production. This up-regulation of glycolytic flux and
glucose transport is now associated with 90% of cancers.In order to
improve the overall understanding of the “Warburg effect” two forms
of systems biology have been implemented - comparative in vitro
analysis of kinetic activities and dynamic modelling. In this
analysis, human breast cancer cell lines MCF-7, MDA-MB-231 and T47D
and a non transformed breast cell line MCF-10A were used to
identify key similarities and differences in kinetic activities
across the glycolytic pathway. Additionally, activities of key
glycolytic enzymes hexokinase, pyruvate kinase and lactate
dehydrogenase were compared under hypoxic conditions to further
understand regulation of cancer cells.The most prominent feature
that arose from comparing the kinetic activities of the three
malignant and one non-malignant cell line is that each cell line
has its own specific set of activities for glycolysis. This
indicates that there are differences in regulation across the
glycolytic pathway for each of these cell lines. This is of
specific interest in the search for therapeutic targets.Further, we
determined that despite the prominence of oncogenic HIF signalling
activities of hexokinase, pyruvate kinase and lactate dehydrogenase
were further modulated by growth under hypoxic conditions.Despite
the lack of obvious distinct kinetic differences between the
non-cancerous and cancerous cells lines some discernible
differences are apparent when modelled in silico.
None
None
Advisors/Committee Members: WILLIAMS, KAYE KJ, Williams, Kaye, Westerhoff, Hans.
Subjects/Keywords: Glycolysis; Enzyme Kinetics; Metabolomics; Mathematical Modelling; Breast Cancer Cell lines
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Chicago ·
MLA ·
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APA (6th Edition):
Blount, K. (2014). Cancer Systems Biology: Is the devil in the glycolytic
detail?. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:222501
Chicago Manual of Style (16th Edition):
Blount, Kathryn. “Cancer Systems Biology: Is the devil in the glycolytic
detail?.” 2014. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:222501.
MLA Handbook (7th Edition):
Blount, Kathryn. “Cancer Systems Biology: Is the devil in the glycolytic
detail?.” 2014. Web. 15 Dec 2019.
Vancouver:
Blount K. Cancer Systems Biology: Is the devil in the glycolytic
detail?. [Internet] [Doctoral dissertation]. University of Manchester; 2014. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:222501.
Council of Science Editors:
Blount K. Cancer Systems Biology: Is the devil in the glycolytic
detail?. [Doctoral Dissertation]. University of Manchester; 2014. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:222501

University of Manchester
2.
Bryant, Jennifer.
Neuroendocrine and epithelial markers of small cell lung
cancer.
Degree: 2015, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:259793
► Small cell lung cancer (SCLC) is an extremely aggressive disease characterized by early metastasis and acquired resistance to therapy. SCLC is distinguished by its neuroendocrine…
(more)
▼ Small cell lung cancer (SCLC) is an extremely
aggressive disease characterized by early metastasis and acquired
resistance to therapy. SCLC is distinguished by its neuroendocrine
(NE) component; the role of which is not fully understood in
metastasis and response to therapy. Patients respond exceptionally
well to first round chemotherapy; however, relapse with
therapy-resistant tumours is virtually inevitable. Hypoxic regions
within tumours can contribute towards metastasis and therapy
resistance, highlighting hypoxia-targeted therapy as a novel
approach for improving treatment for SCLC patients. Tumours are
highly phenotypically heterogeneous, raising debate over the roles
played by each cell type. Analysis of NE and epithelial markers in
SCLC cell lines highlighted this inter-tumour heterogeneity.
Further heterogeneity is displayed in SCLC xenograft tumours that
show areas of dual epithelial and NE marker expression as well as
regions negative for both markers. Irradiating xenograft tumours
enhanced heterogeneity of the NE marker, pro-opiomelanocortin
(POMC), which is ectopically secreted by a subset of SCLC tumours.
Examining changes in marker expression post-therapy could provide
vital information regarding transitions that can serve to guide
therapy. SCLC is a highly metastatic disease. The role of the NE
phenotype in human SCLC is not fully understood, but is considered
essential for metastasis in murine models. Sub-cutaneous,
intravenous and intra-splenic injection were carried out and
resulted in no metastasis, spontaneous tumour generation and
peripheral liver tumour growth, respectively. POMC expression was
present and extremely heterogeneous within the liver, suggesting
that NE properties are maintained in metastases; however, further
work is necessary to develop a more consistent metastatic model
that can be used to assess responses to therapy in a more
clinically relevant setting. SCLC tumours proliferate rapidly and
outgrow their nutrient and oxygen supplies, resulting in hypoxic
conditions. Here, carbonic anhydrase IX (CA IX) becomes
up-regulated in order to maintain pH levels suitable for survival.
The specific CA IX inhibitor, S4, induces hypoxia-specific cell
death in vitro and impairs tumour growth in vivo. This response is
further accentuated by combining S4 with single or repeated
cisplatin doses. Combination treatment reduced gene expression of
S-phase kinase-associated protein (Skp2), associated with cisplatin
resistance. CA IX inhibition combined with cisplatin chemotherapy
therefore presents a novel treatment for SCLC tumours that could
reduce therapy resistance. In summary, heterogeneity is extremely
important when choosing treatment options for SCLC and must be
considered when basing treatment on single biopsies. NE and
epithelial markers are present within sub-cutaneous and liver
tumours; however, a reliable multi-organ metastatic model is
necessary to fully appreciate the role of these markers in the
spread of SCLC. Hypoxic regions within sub-cutaneous xenograft
tumours upregulate CA IX.…
Advisors/Committee Members: WILLIAMS, KAYE KJ, White, Anne, Williams, Kaye.
Subjects/Keywords: small cell lung cancer; Neuroendocrine; Heterogeneity; Metastasis; Xenograft; Hypoxia; Carbonic anhydrase IX
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Bryant, J. (2015). Neuroendocrine and epithelial markers of small cell lung
cancer. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:259793
Chicago Manual of Style (16th Edition):
Bryant, Jennifer. “Neuroendocrine and epithelial markers of small cell lung
cancer.” 2015. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:259793.
MLA Handbook (7th Edition):
Bryant, Jennifer. “Neuroendocrine and epithelial markers of small cell lung
cancer.” 2015. Web. 15 Dec 2019.
Vancouver:
Bryant J. Neuroendocrine and epithelial markers of small cell lung
cancer. [Internet] [Doctoral dissertation]. University of Manchester; 2015. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:259793.
Council of Science Editors:
Bryant J. Neuroendocrine and epithelial markers of small cell lung
cancer. [Doctoral Dissertation]. University of Manchester; 2015. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:259793

University of Manchester
3.
Morris, Olivia Ann.
Development and Application of a Generic Platform for
Radiolabelling Affinity Peptides & Proteins with PET
Isotopes.
Degree: 2017, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:309488
► Positron emission tomography (PET) is a quantitative and non-invasive investigative tool, which permits the identification of pivotal biomarkers and their role in disease onset, transformation…
(more)
▼ Positron emission tomography (PET) is a
quantitative and non-invasive investigative tool, which permits the
identification of pivotal biomarkers and their role in disease
onset, transformation and progression. Quantitative detection of
these disease biomarkers has qualified PET as a pioneering approach
to rapid diagnosis. The tool is a powerful approach to patient
diagnosis and investigation of inter- and intra- patient disease
heterogeneity thereby supporting patient stratification and forging
a truly personalised medicine approach. The sensitivity of PET is
complemented by use of highly selective radiopharmaceuticals; this
has fuelled the popularity of radiolabelled affinity peptides and
proteins (APPs). Inspired by the high selectivity and affinity of
biological protein-protein interactions, APP-based radiotracers are
an increasingly popular class of radiotracer. Monoclonal antibodies
(mAbs) are an important category of APP-based radiotracer; their
prevalence in the field is attributed to their high target
selectivity and affinity characteristics, which, will likely,
uphold their popularity in the field. Yet, advancements in protein
engineering has transformed the landscape of APP-based
radiotracers, indicated by a preference for small radiolabelled
APPs which exhibit rapid pharmacokinetics whilst retaining high
target specificity and affinity. Their fast pharmacokinetics is
matched by the half-life of 18F which has been defined as an ideal
PET radionuclide. 18F APP radiolabelling hasn’t yet met its full
potential due to its inherent challenges; many approaches have been
adopted, ranging from non-site specific radiolabelling with amine
reactive prosthetic groups to site-specific methods including oxime
bond formation. The radiochemistry of 18F APP radiolabelling is
diverse and evolving; yet ideal goals are apparent including
site-specificity, fast reaction kinetics, mild labelling conditions
and applicability to automation. Automation is challenging for
multi-step 18F APP radiolabelling methods, but is a key
developmental step. Automation helps with conformance to good
manufacturing practice (GMP) by enhancing process robustness,
consistency and reliability. It also permits the radiosynthesis of
clinically relevant radiotracer doses. The development of an
automated generic platform, or a method that adopts a general
approach, for APP radiolabelling with 18F would assist in [18F]APPs
meeting their potential in PET and, importantly, aid their
translation from bench-to-bedside.
Advisors/Committee Members: WILLIAMS, KAYE KJ, Williams, Kaye, Mcmahon, Adam.
Subjects/Keywords: 18F prosthetic group; peptide radiolabelling; PET
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Morris, O. A. (2017). Development and Application of a Generic Platform for
Radiolabelling Affinity Peptides & Proteins with PET
Isotopes. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:309488
Chicago Manual of Style (16th Edition):
Morris, Olivia Ann. “Development and Application of a Generic Platform for
Radiolabelling Affinity Peptides & Proteins with PET
Isotopes.” 2017. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:309488.
MLA Handbook (7th Edition):
Morris, Olivia Ann. “Development and Application of a Generic Platform for
Radiolabelling Affinity Peptides & Proteins with PET
Isotopes.” 2017. Web. 15 Dec 2019.
Vancouver:
Morris OA. Development and Application of a Generic Platform for
Radiolabelling Affinity Peptides & Proteins with PET
Isotopes. [Internet] [Doctoral dissertation]. University of Manchester; 2017. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:309488.
Council of Science Editors:
Morris OA. Development and Application of a Generic Platform for
Radiolabelling Affinity Peptides & Proteins with PET
Isotopes. [Doctoral Dissertation]. University of Manchester; 2017. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:309488
4.
Rowling, Emily.
Pre-clinical evaluation of novel anti-metastatic
targets.
Degree: 2014, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:240986
► Background: Radiotherapy is used in the treatment of over 50% of cancer patients and bar surgery, is the most effective cancer intervention. However, in the…
(more)
▼ Background: Radiotherapy is used in the treatment
of over 50% of cancer patients and bar surgery, is the most
effective cancer intervention. However, in the clinic secondary
malignancies have been observed following radiotherapy and in vitro
increased cell migration and invasion have been seen following
radiation. The Src/FAK signalling pathway is known to play an
important role in the metastatic phenotype through its involvement
in cell adhesion, migration and invasion and we have previously
demonstrated that radiotherapy can activate this pathway along with
the phosphoinositide 3-kinase (PI3K) pathway, also associated with
tumour metastases and an aggressive phenotype. Using
pharmacological inhibitors, we have investigated combination
approaches to evaluate whether Src and PI3K targeting is beneficial
in a radiotherapy context, especially focusing on metastatic
phenotype. We wished to relate pathway activation to cellular
phenotype and increase understanding of the metastatic cascade, the
processes involved and the signalling pathways taking the
lead.Method: Using thyroid carcinoma cell lines FTC133 and 8505c
the effects of Src inhibition using AZD0530, FAK inhibition using
FAKi and PI3K inhibition using GDC-0941 were studied. The effects
of radiotherapy alone, and in combination with the above
inhibitors, were also studied. In vitro MTT, apoptosis and
clonogenic assays were used to assess cell proliferation and cell
survival and scratch assays, cell adhesion and cell spreading
assays were used to assess the effects of the drugs on metastatic
characteristics. In vivo tumour growth, survival and ex vivo
clonogenics were used to measure the effects of AZD0530 and
GDC-0941. Western blotting, immunofluorescence and
immunohistochemistry was used to observe the effects on pathway
activation and protein localisation.Results: Src and FAK inhibition
reduced metastatic characteristics of thyroid carcinoma cell lines
in vitro such as cell spreading and migration. FAK inhibition
showed a greater effect on cell survival by MTT, clonogenic and
apoptosis. In the thyroid carcinoma cell lines radiotherapy
enhanced the metastatic phenotype. This was seen by enhanced
activation of the Src and PI3K pathways, increased migration and
invasion in vitro and enhanced tumour metastasis in vivo. By
combining Src inhibition with radiation a reduction in metastatic
characteristics was observed and by combining PI3K inhibition with
radiotherapy radiosensitivity could be improved. With the triple
combination of Src and PI3K inhibition with radiotherapy a
significant reduction in cell survival was demonstrated in vitro
compared to radiation alone and either inhibitor combined with
radiation, with a corresponding significant reduction in tumour
growth being observed in vivo. With the combination of Src and PI3K
inhibition significant reductions in metastatic characteristics
were also observed both in vitro and in vivo seen by a reduction in
cell migration and tumour metastasis. Finally combined inhibition
of the Src and PI3K pathway reduced the…
Advisors/Committee Members: Williams, Kaye.
Subjects/Keywords: Src; metastasis; Radiotherapy; PI3K; Thyroid
…related rights in it (the “Copyright”) and she has given The University
of Manchester…
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Rowling, E. (2014). Pre-clinical evaluation of novel anti-metastatic
targets. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:240986
Chicago Manual of Style (16th Edition):
Rowling, Emily. “Pre-clinical evaluation of novel anti-metastatic
targets.” 2014. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:240986.
MLA Handbook (7th Edition):
Rowling, Emily. “Pre-clinical evaluation of novel anti-metastatic
targets.” 2014. Web. 15 Dec 2019.
Vancouver:
Rowling E. Pre-clinical evaluation of novel anti-metastatic
targets. [Internet] [Doctoral dissertation]. University of Manchester; 2014. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:240986.
Council of Science Editors:
Rowling E. Pre-clinical evaluation of novel anti-metastatic
targets. [Doctoral Dissertation]. University of Manchester; 2014. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:240986
5.
Kinnersley, Janet.
Evaluation of novel anti-metastatic therapy.
Degree: 2011, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:141162
► The occurrence of metastasis from the primary tumour to distant organs is the primary cause of cancer mortality and therefore a highly attractive therapeutic target…
(more)
▼ The occurrence of metastasis from the primary
tumour to distant organs is the primary cause of cancer mortality
and therefore a highly attractive therapeutic target to improve
cancer survival rates. Targeted anti-metastatic therapies hold the
potential for lower cancer mortality rates, lower rates of
reoccurrence and better quality of life for cancer patients. Src is
a non receptor tyrosine kinase, and the first recognized oncogene,
which is known to play a role in metastasis. Increased activity of
SFKs has been found in many human tumours, correlating with
invasiveness and poor prognosis, and in experimental tumour cell
lines, contributing to enhanced cell migration and invasion,
adhesion independent growth, survival and proliferation. Despite a
body of evidence in pre-clinical trials demonstrating that
inhibition of Src reduces the occurrence of cellular events
associated with metastasis in vitro, and reduces tumour metastasis
to distant organs in in vivo models, efficacy has not translated to
phase II clinical trials. One such recent failure has been a phase
II trial with the Src inhibitor AZD0530 in recurrent advanced or
metastatic soft tissue sarcoma. Here we investigate the
anti-metastatic effects of Src inhibition in the HT1080
fibrosarcoma cell in vitro in order to elucidate further the
effects of SFK inhibition in sarcoma cells laying the ground work
for future in vivo work investigating potential compensatory
mechanism overcoming SFK inhibition in sarcoma cells. We found that
SFK inhibition of HT1080 cells with AZD0530 inhibited cell
migration and spreading but had no effect on cell polarization.
AZD0530 treatment caused active paxillin (Tyr31-p) to relocalize
from focal adhesions to the cytoplasm but had no effect on staining
of active FAK (Tyr861-p). Paradoxically AZD0530 treatment led to
increased phosphorylation of Src at the negative regulatory site
Tyr 530. Results seen here provide evidence to warrant further
elucidation of the effects of Src inhibition in HT1080 cells in
order to elucidate potential combination therapies and biomarkers
of tumour sensitivity to Src inhibition.
Advisors/Committee Members: Williams, Kaye.
…University of
Manchester).
2.3 Treatments
The src inhibitor, AZD0530 was supplied by Astra…
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Kinnersley, J. (2011). Evaluation of novel anti-metastatic therapy. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:141162
Chicago Manual of Style (16th Edition):
Kinnersley, Janet. “Evaluation of novel anti-metastatic therapy.” 2011. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:141162.
MLA Handbook (7th Edition):
Kinnersley, Janet. “Evaluation of novel anti-metastatic therapy.” 2011. Web. 15 Dec 2019.
Vancouver:
Kinnersley J. Evaluation of novel anti-metastatic therapy. [Internet] [Doctoral dissertation]. University of Manchester; 2011. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:141162.
Council of Science Editors:
Kinnersley J. Evaluation of novel anti-metastatic therapy. [Doctoral Dissertation]. University of Manchester; 2011. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:141162

University of Manchester
6.
Henderson, Fiona Rae.
Mass Spectrometry Imaging of Lipid Profiles in
Disease.
Degree: 2017, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:308361
► It is well established that lipids play an important role in diseases such as non-alcoholic fatty liver disease and cardiovascular diseases. However, in the past…
(more)
▼ It is well established that lipids play an
important role in diseases such as non-alcoholic fatty liver
disease and cardiovascular diseases. However, in the past decade,
it has come to light that lipids may be important in other
diseases; particularly in cancer and neurological disorders. Here,
lipid metabolism has been investigated using pre-clinical cancer
models for melanoma, glioma, non-small-cell lung cancer and
colorectal cancer. The role of lipids in the recovery post-stroke
has also been studied. Mass spectrometry imaging offers an ideal
tool to study lipids in tissue ex-vivo. Lipids ionise well in a
number of mass spectrometry modalities, and hundreds of lipids can
be imaged in one mass spectrometry imaging experiment. Furthermore,
mass spectrometry imaging offers excellent spatial resolution. In
this work, both MALDI-MS and DESI-MS have been used for mass
spectrometry imaging.Tumour lipid heterogeneity has been a
particular focus of this this project. Heterogeneity exists within
tumours, as well as between tumours in the same patient; and this
causes major problems for therapy. Owing to the untargeted nature,
and high spatial resolution of mass spectrometry imaging, it is an
excellent technique to study lipid heterogeneity. Adjacent sections
(or in some cases the same section used for mass spectrometry
imaging), were used for immunofluorescence and H&E staining. By
comparing mass spectrometry images with staining techniques,
biological reasons for lipid heterogeneity can be established.
Here, a particular focus has been on hypoxia (low oxygen tensions),
which is a key
contributor to tumour heterogeneity, and is
associated with aggressive cancers. Additionally, hypoxia is a
feature of ischaemic stroke, and lipids in ischaemic stroke have
also been investigated. PET is a non-invasive imaging technique
which is able to image a radiolabelled molecule (tracer) in the
body. Here, PET has been used as a complementary in-vivo technique
to mass spectrometry imaging. The tracers [11C] acetate and
[18F]-FTHA have been used to image fatty acid synthase and fatty
acid uptake in tumours; both of which are hypothesised to be key in
cancer progression. REIMS is a newly established mass spectrometry
technique. It is ideal for analysing lipids in cells, as sample
preparation is minimal. Here, approaches for cell pellet analysis
have been tested, and used to detect lipids in cancer cell
lines.
Advisors/Committee Members: MCMAHON, ADAM AW, Williams, Kaye, Mcmahon, Adam.
Subjects/Keywords: Mass spectrometry imaging; Lipids; Cancer
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Henderson, F. R. (2017). Mass Spectrometry Imaging of Lipid Profiles in
Disease. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:308361
Chicago Manual of Style (16th Edition):
Henderson, Fiona Rae. “Mass Spectrometry Imaging of Lipid Profiles in
Disease.” 2017. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:308361.
MLA Handbook (7th Edition):
Henderson, Fiona Rae. “Mass Spectrometry Imaging of Lipid Profiles in
Disease.” 2017. Web. 15 Dec 2019.
Vancouver:
Henderson FR. Mass Spectrometry Imaging of Lipid Profiles in
Disease. [Internet] [Doctoral dissertation]. University of Manchester; 2017. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:308361.
Council of Science Editors:
Henderson FR. Mass Spectrometry Imaging of Lipid Profiles in
Disease. [Doctoral Dissertation]. University of Manchester; 2017. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:308361
7.
Leung, Travis.
The regulation of CYP2E1 gene expression and its role in
breast cancer.
Degree: 2013, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:188013
► Cytochromes(CYP) P450 are class of heme-containing enzymes involved in Phase I metabolism of a large number of xenobiotics. The CYP family member CYP2E1 is involved…
(more)
▼ Cytochromes(CYP) P450 are class of heme-containing
enzymes involved in Phase I metabolism of a large number of
xenobiotics. The CYP family member CYP2E1 is involved in the
metabolism of many xenobiotics and procarcinogens including
ethanol, acetone, nitrosamine, pyridine, isoniazid and carbon
tetrachloride (CCl4), which are also CYP2E1-inducing agents. CYP2E1
comprises approximately 7% of the liver CYP content and it is also
expressed in kidney, lung, brain, gastrointestinal tract and breast
tissue implying that this enzyme is implicated in other biological
processes aside from its role in Phase I metabolism. Several
studies converge to the conclusion that CYP2E1 is induced under
many pathological conditions including cancer, obesity, and type 2
diabetes. Increased hepatic ketogenesis and insulin resistance are
the possible mechanisms mediating CYP2E1 induction in these
conditions. Elevated CYP2E1 expression has been reported in the
presence of proinflammatory cytokines such as interleukin 6 (IL-6)
and tumor necrosis factor alpha (TNF-α). CYP2E1 generates the
highest level of reactive oxygen species (ROS) among the CYP450
superfamily and transient overexpression of this CYP family member
in COS cells increases the production of mitochondrial ROS, even in
the absence of substrate. Oxidative stress induced by CYP2E1
disturbs the folding capacity of the endoplasmic reticulum (ER),
with concomitant alterations in the mRNA and protein expression of
the ER stress proteins GRP78 and GRP94. Initially the unfolded
protein response (UPR) restores ER homeostasis, and cell viability,
but at later stages the accumulation of unfolded proteins
stimulates pro-apoptotic signals. Cell death induced by ER stress
has been reported in several conditions including hypoxia, and
diseases such as diabetes and heart disease. To gain better
understanding of the factors regulating CYP2E1 gene expression in
breast cancer cells we studied CYP2E1 mRNA and protein levels in
MCF7 (p53wt) and MDA-MB-231 (p53 mutated) breast cancer cells and
identified that CYP2E1 was under p53 and HIF-1α transcriptional
control in both of these cell lines treated with etoposide or
desferrioxamine respectively. In addition, CYP2E1 was
differentially expressed in the low metastatic potential MCF7 cell
line compared to highly metastatic MDA-MB-231 cells in accord with
clinical studies indicating that CYP2E1 isoenzyme is expressed in
lower levels in patients at clinical stages II, III, and IV which
exhibit higher metastatic potential than in patients at stage I of
breast cancer. To further investigate the functional significance
of the difference in CYP2E1 levels the generation of ROS in breast
cancer cells ectopically overexpressing CYP2E1 or in cells in which
CYP2E1 expression had been silenced was assessed. Our results
indicated that CYP2E1 overexpression resulted in higher ROS
generation, which coincided with increased autophagy biomarker
expression, increased endoplasmic reticulum stress, detected by
XBP1 mRNA splicing in fluorescent reporter assays, and inhibition…
Advisors/Committee Members: WILLIAMS, KAYE KJ, Williams, Kaye, Demonacos, Constantinos.
…any copyright in it (the “Copyright”) and s/he has given The University of… …Manchester the right to use such Copyright for any administrative, promotional,
educational and/or…
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Leung, T. (2013). The regulation of CYP2E1 gene expression and its role in
breast cancer. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:188013
Chicago Manual of Style (16th Edition):
Leung, Travis. “The regulation of CYP2E1 gene expression and its role in
breast cancer.” 2013. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:188013.
MLA Handbook (7th Edition):
Leung, Travis. “The regulation of CYP2E1 gene expression and its role in
breast cancer.” 2013. Web. 15 Dec 2019.
Vancouver:
Leung T. The regulation of CYP2E1 gene expression and its role in
breast cancer. [Internet] [Doctoral dissertation]. University of Manchester; 2013. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:188013.
Council of Science Editors:
Leung T. The regulation of CYP2E1 gene expression and its role in
breast cancer. [Doctoral Dissertation]. University of Manchester; 2013. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:188013
8.
Telfer, Brian.
Application of the dorsal window chamber to tumour
vasculature manipulation studies.
Degree: 2012, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:156044
► Developing and applying pre-clinical tumour models in order to determine the mechanistic action of applied therapies is essential if we aim to improve antitumour strategies…
(more)
▼ Developing and applying pre-clinical tumour models
in order to determine the mechanistic action of applied therapies
is essential if we aim to improve antitumour strategies in the
clinical setting. The chaotic nature of tumour vasculature impacts
directly on the effectiveness of combined chemo-radiotherapy and
antiangiogenic (AA) strategies and as such warrants closer study.
This work looked at the effects of novel AAs combined with
clinically relevant radiotherapy (RT) using both conventional
murine xenograft growth delay studies and real-time imaging. The
imaging methodology was the non-invasive Dorsal Window
Chamber/Intra Vital Microscopy (DWC/IVM) model which allows the
study of real-time vascular responses to these therapies. The
DWC/IVM model was applied to determine whether the DNA repair
inhibitors Nicotinamide, AG14361 and AGO14699 had additional modes
of action which could contribute to tumour
radioresistance/radiosensitivity. Using the DWC/IVM model a
secondary quantifiable mechanistic function was determined where
these drugs also increased tumour vessel permeability. The DWC/IVM
model was also used to investigate the effects of AZD2171 and
AZD6244 combined with radiotherapy. These agents can inhibit
angiogenic signalling pathways and it was demonstrated that both
drugs worked by reducing tumour microvascular density when used in
combination with radiation. In studies looking at the influence of
hypoxia inducible factor-1(HIF-1) on tumour response to radiation
the DWC/IVM model provided measurable differences in the
microvascular density between HIF-1 deficient and HIF-1 competent
tumours. The DWC/IVM model allowed the direct visualisation and
quantification of the less well developed vasculature in HIF-1
deficient tumours compared to that found in HIF-1 competent
tumours. The results provide a mechanistic basis for understanding
the improved response to radiotherapy of HIF-1 deficient tumours.By
applying the DWC/IVM model to conventional murine xenograft models
the DWC/IVM proved itself as a useful research tool where
continuous real-time non-invasive measurements could be made
without the need for large numbers of time points or laborious
histological analysis.
Advisors/Committee Members: WILLIAMS, KAYE KJ, Stratford, Ian, Williams, Kaye.
Subjects/Keywords: Tumour vascular dorsal window radiation
model
…rights in it (the ―Copyright‖) and s/he has given The
University of Manchester… …his work at the University of Manchester in the Department of Anatomy
and trained as a…
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Telfer, B. (2012). Application of the dorsal window chamber to tumour
vasculature manipulation studies. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:156044
Chicago Manual of Style (16th Edition):
Telfer, Brian. “Application of the dorsal window chamber to tumour
vasculature manipulation studies.” 2012. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:156044.
MLA Handbook (7th Edition):
Telfer, Brian. “Application of the dorsal window chamber to tumour
vasculature manipulation studies.” 2012. Web. 15 Dec 2019.
Vancouver:
Telfer B. Application of the dorsal window chamber to tumour
vasculature manipulation studies. [Internet] [Doctoral dissertation]. University of Manchester; 2012. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:156044.
Council of Science Editors:
Telfer B. Application of the dorsal window chamber to tumour
vasculature manipulation studies. [Doctoral Dissertation]. University of Manchester; 2012. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:156044
9.
Burke, Daniel.
Development of Robust Multimodal Imaging Biomarkers of
the Non-viable Tumour Component for Cancer Drug
Development.
Degree: 2013, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:204162
► This project was initiated with the objective of obtaining a magnetic resonance imaging (MRI) derived measure of the non-viable cell fraction of a tumour. The…
(more)
▼ This project was initiated with the objective of
obtaining a magnetic resonance imaging (MRI) derived measure of the
non-viable cell fraction of a tumour. The word “biomarker” pervades
much of the quantitative imaging literature. There exist a variety
of imaging parameters which purport to convey information about the
underlying pathophysiology. These are generically labelled as
“biomarkers”, although a review of the literature indicates that
there is little, (or often contradictory) evidence to support this
assertion. Indeed, the absence of robust validation is in evidence
across the broad spectrum of imaging modalities. As the development
of novel chemotherapeutic agents progresses from in vitro studies
performed in a Petri dish to in vivo animal experiments, treatment
efficacy can be gauged by a number of methods, one of which is
imaging. Translational imaging offers the opportunity of developing
and validating imaging techniques in the preclinical setting, with
potential for their adoption into a clinical trial. The Apparent
Diffusion Coefficient (ADC) of water proton spins has emerged as a
potential “biomarker” of cell death. Two studies were performed to
investigate the relationship between ADC and the pathophysiologic
changes occurring subsequent to radiotherapy or chemotherapy in
preclinical cancer models. Dynamic contrast enhanced magnetic
resonance imaging (DCE-MRI) is purported to offer a means of
quantifying the non-viable cell fraction through the
pharmacokinetic parameter “ve”, the extravascular extracellular
space volume per unit volume of tissue. Performing a DCE-MRI
experiment requires measurements of T1 before and after the
administration of a contrast agent. To this end, methods
development undertaken included extensive validation experiments to
determine the accuracy and reproducibility of the T1 relaxation
time for use in a DCE-MRI study. Validation of ADC measurements was
performed through the use of a novel ice water phantom designed for
a narrow bore preclinical MRI scanner. A means of determining the
quantity of contrast agent within a tumour can be performed through
the use of ICP-MS (Inductively Coupled Plasma Mass Spectrometry). A
validation experiment was performed through the preparation of a
phantom of known quantity of contrast agent and comparison against
that returned from ICP-MS analysis. This thesis contends that the
development of an MRI derived measure of the non-viable cell
fraction does not appear viable within the framework of the present
pharmacokinetic models and imaging hardware available. The
literature fails to address significant confounds in the scientific
method, indeed this is in evidence in the variability of observed
imaging parameters.
This project was initiated with the objective of
obtaining a magnetic resonance imaging (MRI) derived measure of the
non-viable cell fraction of a tumour. The word “biomarker” pervades
much of the quantitative imaging literature. There exist a variety
of imaging parameters which purport to convey information about the
underlying…
Advisors/Committee Members: WILLIAMS, KAYE KJ, O'CONNOR, JAMES JP, Williams, Kaye, O'Connor, James, Waterton, John.
Subjects/Keywords: Magnetic resonance imaging; Apparent diffusion coefficient; Longitudinal relaxation time
…observed imaging parameters.
Institution : The University of Manchester
Candidate : Daniel Peter… …x28;the “Copyright”) and s/he has given the University of Manchester certain rights to…
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Burke, D. (2013). Development of Robust Multimodal Imaging Biomarkers of
the Non-viable Tumour Component for Cancer Drug
Development. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:204162
Chicago Manual of Style (16th Edition):
Burke, Daniel. “Development of Robust Multimodal Imaging Biomarkers of
the Non-viable Tumour Component for Cancer Drug
Development.” 2013. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:204162.
MLA Handbook (7th Edition):
Burke, Daniel. “Development of Robust Multimodal Imaging Biomarkers of
the Non-viable Tumour Component for Cancer Drug
Development.” 2013. Web. 15 Dec 2019.
Vancouver:
Burke D. Development of Robust Multimodal Imaging Biomarkers of
the Non-viable Tumour Component for Cancer Drug
Development. [Internet] [Doctoral dissertation]. University of Manchester; 2013. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:204162.
Council of Science Editors:
Burke D. Development of Robust Multimodal Imaging Biomarkers of
the Non-viable Tumour Component for Cancer Drug
Development. [Doctoral Dissertation]. University of Manchester; 2013. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:204162
10.
Kotze, Helen.
Systems Biology of Chemotherapy in Hypoxia
Environments.
Degree: 2012, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:182472
► Introduction: Hypoxia is found in solid cancerous tumours. The presence of hypoxia within tumours inhibits anti-cancer treatment strategies such as chemotherapy from being completely effective…
(more)
▼ Introduction: Hypoxia is found in solid cancerous
tumours. The presence of hypoxia within tumours inhibits
anti-cancer treatment strategies such as chemotherapy from being
completely effective and it is suspected that multiple mechanisms
contribute to the resistance. Methods: In this project a systems
biology approach was applied to determine how the toxicity of
doxorubicin is affected by hypoxia at the metabolome level. A
multitude of analytical techniques were applied to analyse the
intracellular metabolism of a monolayer of cancer cells
(MDA-MB-231). Metabolic profiling was used to determine metabolite
markers related to hypoxia-induced chemoresistance. For this gas
chromatography mass spectrometry (GC-MS) and ultra high performance
liquid chromatography mass spectrometry (UHPLC-MS) were used.
Furthermore, network-based correlation analysis was developed as a
novel tool to bridge the gap between metabolomics dataset and
systems biology modelling. This methodology was applied to
elucidate novel metabolic pathways as potential therapeutic targets
to overcome hypoxia-induced chemoresistance. This algorithm
determines significant correlation differences between different
physiological states, and through applying graph-theory on large
genome scale models; it is possible to construct a metabolic
network of the pathways connecting the pair-wise correlation.
Finally, imaging mass spectrometry using time-of-flight secondary
ion mass spectrometry (ToF-SIMS) was developed as a tool for in
situ metabolite analysis to investigate the metabolic response to
chemotherapy in multi-tumour spheroids (MTSs). Results: Metabolic
fingerprinting analysis characterised a snapshot of cells exposed
to various environmental perturbations. Metabolite markers
associated with hypoxia-induced chemoresistance were related to
metabolic pathways including gluconeogenesis, DNA synthesis and
fatty acid synthesis. Furthermore, network-based correlation
analysis revealed specific metabolites in the fatty acid synthesis
pathways were contributing to drug resistance, which included
malonyl-CoA, 3-oxoeicosanoyl-CoA, stearoyl-CoA and octadecanoic
acid. To facilitate the detection of metabolites in ToF SIMS
datasets, a series of metabolites standard spectra were acquired.
Hypoxic metabolite markers detected in ToF-SIMS data of cell
lysates included glycine, lactic acid and succinic acid, which were
also shown to be metabolite markers in GC-MS metabolic data.
Furthermore, MTS sections were imaged using ToF-SIMS to profile the
chemical response to chemotherapy treatment within the oxygen
gradient. Loadings from image PCA were explored to determine the
metabolic response in the highly oxygenated outer region and
hypoxic inner region of the MTS. Conclusion: A multitude of
analytical techniques were able to contribute to elucidating the
metabolic mechanisms associated with hypoxia-induced
chemoresistance. Metabolic profiling combined with a systems
biology approach was further able to identify potential underlying
metabolic regulation of resistance. Finally…
Advisors/Committee Members: GOODACRE, ROY R, WILLIAMS, KAYE KJ, MCMAHON, ADAM AW, Lockyer, Nicholas, Goodacre, Roy, Williams, Kaye, Mcmahon, Adam, Westerhoff, Hans.
Subjects/Keywords: Metabolomics; Hypoxia; ToF-SIMS; Chemotherapy; Systems biology
…81
14
The University of Manchester
Faculty of Engineering and Physical Sciences
ABSTRACT… …x29; and s/he has given
The University of Manchester certain rights to use such Copyright…
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Kotze, H. (2012). Systems Biology of Chemotherapy in Hypoxia
Environments. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:182472
Chicago Manual of Style (16th Edition):
Kotze, Helen. “Systems Biology of Chemotherapy in Hypoxia
Environments.” 2012. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:182472.
MLA Handbook (7th Edition):
Kotze, Helen. “Systems Biology of Chemotherapy in Hypoxia
Environments.” 2012. Web. 15 Dec 2019.
Vancouver:
Kotze H. Systems Biology of Chemotherapy in Hypoxia
Environments. [Internet] [Doctoral dissertation]. University of Manchester; 2012. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:182472.
Council of Science Editors:
Kotze H. Systems Biology of Chemotherapy in Hypoxia
Environments. [Doctoral Dissertation]. University of Manchester; 2012. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:182472
11.
Armitage, Emily Grace.
Systems Biology of HIF Metabolism in Cancer.
Degree: 2012, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:177044
► AbstractThe University of ManchesterFaculty of Engineering and Physical SciencesAbstract of thesis entitled ‘Systems Biology of HIF Metabolism in Cancer’Submitted by Emily Grace Armitage for the…
(more)
▼ AbstractThe
University of ManchesterFaculty of
Engineering and Physical SciencesAbstract of thesis entitled
‘Systems Biology of HIF Metabolism in Cancer’Submitted by Emily
Grace Armitage for the degree of Doctor of Philosophy, September
2012Cancer is one of the most devastating human diseases that cause
a vast number of mortalities worldwide each year. Cancer research
is one of the largest fields in the life sciences and despite many
astounding breakthroughs and contributions over the past few
decades, there is still a considerable amount to unveil on the
function of cancer that would improve diagnostics, prognostics and
therapy. Since cancer is known to involve a wide range of
processes, applying methods to study it from a systems perspective
could reveal new properties of cancer. Systems biology is becoming
an increasingly popular tool in the life sciences. The approach has
been applied to many biological and biomedical analyses drawing
upon recent advancements in technology that make high throughput
analyses of samples and computational modelling possible. In this
thesis, the effect of hypoxia inducible factor-1 (HIF-1) on cancer
metabolism, the entity considered most closely related to phenotype
has been investigated. This transcription factor is known to
regulate a multitude of genes and proteins to promote survival in a
low oxygen environment that is prevalent in solid tumours. However
its effect on the metabolome is less well characterised. By
revealing the effect of HIF-1 on the metabolome as a system it is
hoped that phenotypic signatures, key metabolic pathways indicative
of cancer function and potential targets for future cancer therapy,
can be revealed.The system has been studied using two cell models:
mouse hepatocellular carcinoma and human colon carcinoma, whereby
metabolism has been profiled using a range of analytical platforms.
In each model, wild type cells have been compared to cells
deficient in HIF-1 to reveal its effect on cellular metabolism. Gas
chromatography mass spectrometry (GC MS) and ultra high performance
liquid chromatography - mass spectrometry (UHPLC MS) have been
employed for metabolic profiling of cells exposed to a range of
oxygen conditions. Additionally, time-of-flight secondary ion mass
spectrometry (ToF SIMS) has been employed for imaging mass
spectrometric analysis of multicellular tumour spheroids cultured
from wild type cells and cells with dysfunctional HIF-1 to
represent small initiating tumours. Using these techniques in
metabolic profiling it has been possible to reveal metabolites
associated with the effect of oxygen and HIF-1 on cancer metabolism
along with key pathways and hubs that could be targeted in future
therapy. Using imaging mass spectrometry it has been possible to
localise metabolites in situ revealing how tumour structure relates
to function. Finally, a novel approach to consider how metabolites
are correlated with one another in the response to oxygen level or
presence or absence of functional HIF-1 has been undertaken to
better understand the systems…
Advisors/Committee Members: LOCKYER, NICHOLAS NP, GOODACRE, ROY R, MCMAHON, ADAM AW, WESTERHOFF, HANS HV, Lockyer, Nicholas, Goodacre, Roy, Williams, Kaye, Mcmahon, Adam, Westerhoff, Hans.
Subjects/Keywords: Cancer; Systems Biology; Metabolomics; Mass Spectrometry
…130
11
Abstract
The University of Manchester
Faculty of Engineering and Physical Sciences… …given The University of Manchester certain rights to use such Copyright,
including for… …Acknowledgements
I would firstly like to thank my supervisors at The University of Manchester. They… …Biology Doctoral
Training Centre student here at the University of Manchester. Together Helen… …Doctoral Training Centre for Integrative Systems Biology in
The University of Manchester in the…
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Armitage, E. G. (2012). Systems Biology of HIF Metabolism in Cancer. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:177044
Chicago Manual of Style (16th Edition):
Armitage, Emily Grace. “Systems Biology of HIF Metabolism in Cancer.” 2012. Doctoral Dissertation, University of Manchester. Accessed December 15, 2019.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:177044.
MLA Handbook (7th Edition):
Armitage, Emily Grace. “Systems Biology of HIF Metabolism in Cancer.” 2012. Web. 15 Dec 2019.
Vancouver:
Armitage EG. Systems Biology of HIF Metabolism in Cancer. [Internet] [Doctoral dissertation]. University of Manchester; 2012. [cited 2019 Dec 15].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:177044.
Council of Science Editors:
Armitage EG. Systems Biology of HIF Metabolism in Cancer. [Doctoral Dissertation]. University of Manchester; 2012. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:177044
.