Two Ph.D. Student Positions in Cancer Biology (Single Cell Imaging, AMPK
signalling and Bioenergetics) available starting October 2014
Location: Centre for Systems Medicine (CSM) and Department of Physiology and Medical
Physics, Royal College of Surgeons in Ireland (RCSI)
Supervisor: Prof Jochen Prehn
Project background and description:
The PhD student positions will be integrated into the Science Foundation Ireland-funded research group of Prof Jochen
Prehn. Cancer cells have an altered energy metabolism. This may represent an alternative target for therapeutic
intervention. Emerging evidence suggests that alterations in cellular bioenergetics may critically contribute to evasion
of therapeutically-induced, genotoxic cell death. Data from our group suggested that cancer cells are able to restore
energetic and ionic homeostasis even after the successful activation of cell death pathways. Our research and others
has also shown that cellular bioenergetics influence the expression and activation of cell death-regulating proteins
(BCL-2 family proteins), with the kinase, AMPK-activated protein kinase (AMPK), playing a key role in this process. The
SFI-funded project seeks to identify, test, and validate new predictive and prognostic biomarkers for the treatment of
CRC deriving from an integrated, validated, systems-based analysis of cellular bioenergetics, and their dependence on
the mutational status of the tumour. The group has developed key systems models of apoptosis and bioenergetics,
and is one of the few laboratories world-wide that has begun to translate these models into clinical settings.
Position 1:
Single-cell imaging of cellular bioenergetics in KRAS and p53 mutant colon cancer cells: This project will determine
cellular bioenergetics and metabolic signatures under baseline conditions and after induction of apoptosis and their
dependence of the mutational status using single cell imaging. We will use FRET-based indicators of intracellular and
intramitochondrial ATP content), intracellular lactate as well as intracellular glucose. Using pharmacological inhibitors
of bioenergetics pathways, gene silencing of emerging key enzymes and by altering metabolic substrates (glucose,
glutamine, pyruvate, lactate) these experiments will determine in ‘real-time’ the metabolic flux in colon cancer cells
under baseline conditions and during exposure to 5-FU/Oxaliplatin and BCL-2 antagonists.
Position 2:
Effects of AMPK signalling on colon cancer cells survival and growth: This project will characterise the role of AMPK
as a master switch of cell survival and will use a lentivirus to alter the levels of AMPKalphs1/2, the catalytic subunits, in
colon cancer cells harbouring KRAS or p53 mutations. The effects of altering AMPK signalling on cellular bioenergetics
as well as apoptosis susceptibility and cell proliferation in response to 5-FU/Oxaliplatin will be assessed by single cell
microscopy/microfluidics, metabolite analysis, flow cytometry (Annexin V and PI uptake as apoptosis/necrosis
markers; Ki67 staining as proliferation marker), and clonogenic survival assays.
The research infrastructure of the Centre includes a team of cancer biologists, systems biologists, clinical researchers,
and preclinical animal researchers as well as state-of-the-art instrumentation including confocal microscopy and
single-cell FRET analysis, RPPA and protein arrays, Cellomics, high throughput Flow Cytometry, two-photon intravital
microscopy and small animal bioluminescence and PET/CT facilities.
Person Specification:
The candidate should have a B.Sc. or Master degree in Biochemistry, Cell Biology, or a related field. (S)he needs to be
able to independently plan and conduct the research project and to integrate into an interdisciplinary research
environment.
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Stipend:
The successful candidates will receive a tax-free stipend of €18.000 per annum. The positions are funded for three
years.
Application procedure:
Please send a CV and accompanying documentation (certificates, recommendations/references)
to [email protected]
Closing date: Applications should be received by July 31st, 2014.
Relevant publications from the group:
Concannon CG, et al. (2010) AMP kinase-mediated activation of the BH3-only protein Bim couples energy depletion to
stress-induced apoptosis. J Cell Biol. 189: 83-94.
Huber HJ, Dussmann H, Kilbride SM, Rehm M, Prehn JH. Glucose metabolism determines resistance of cancer cells to
bioenergetic crisis after cytochrome-c release. Mol Syst Biol. 2011 Mar 1;7:470.
Kilbride SM, et al. (2010) AMP-activated protein kinase mediates apoptosis in response to bioenergetic stress through
activation of the pro-apoptotic Bcl-2 homology domain-3-only protein BMF. J Biol Chem. 285: 36199-206
Kilbride SM, Prehn JH. Central roles of apoptotic proteins in mitochondrial function. Oncogene. 2013 May
30;32(22):2703-11.
Lindner AU, Concannon CG, Boukes GJ, Cannon MD, Llambi F, Ryan D, Boland K, Kehoe J, McNamara DA, Murray F, Kay
EW, Hector S, Green DR, Huber HJ, Prehn JH. Systems analysis of BCL2 protein family interactions establishes a model
to predict responses to chemotherapy. Cancer Res. 2013 Jan 15;73(2):519-28.
Schmid J, Dussmann H, Boukes GJ, Flanagan L, Lindner AU, O'Connor CL, Rehm M, Prehn JH, Huber HJ. Systems analysis
of cancer cell heterogeneity in caspase-dependent apoptosis subsequent to mitochondrial outer membrane
permeabilization. J Biol Chem. 2012
Note: this Job Description may be subject to change to reflect the evolving requirements of the Department
and RCSI in developing healthcare leaders who make a difference worldwide.
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