Download IRIC NEXT GENERATION INTERNSHIP AWARDS

IRIC NEXT GENERATION
INTERNSHIP AWARDS
2013 EDITION
INTERNSHIP OFFERS
IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
INTERNSHIP OFFERS
Internship #1: Understanding the molecular controls enabling the initiation and the completion of mitosis............................................... 3
Internship #2: β-arrestin-dependent regulation of NF-kB signalling by G protein-coupled receptors .......................................................... 4
Internship #3: ERM regulation during cell division ......................................................................................................................................... 5
Internship #4: Regulation of cell division by mitotic kinases: Implications for cancer development ............................................................. 6
Internship #5: Generation of Drosophila mutants for candidate Rab11-GAP proteins ................................................................................. 7
Internship #6: SCL and LMO1 oncogenes disrupt thymocyte development ................................................................................................... 8
Internship #7: Evaluation of chemical inhibitors of motor proteins in vitro and in live cells ......................................................................... 9
Internship #8: Characterization of a mitotic motor protein (kif14) that is linked to cancer progression ..................................................... 10
Internship #9: Characterization of the activity of the tumor suppressor PAR-4/LKB1 in cell polarity and asymmetric cell division. ........... 11
Internship #11: Complementary roles of BAF60A and BAF60B during the lymphoid vs myeloid differentiation ........................................ 13
Internship #12: Quantitative imaging of chromosome dynamics................................................................................................................. 14
Internship #13: Discovering the early events of cell shape change during physical cell division ................................................................. 15
Internship #14: Production de souches transgéniques de C. elegans à l’aide de transgènes fluorescents .................................................. 16
Internship #15: Mechanisms of action of full antiestrogens in breast tumor cells ...................................................................................... 17
Internship #16: Studying the structure of micro-RNA whose genes are involved in some cancers ............................................................. 18
Internship #17: Discovering better targets for immunotherapy cancer ....................................................................................................... 19
Internship #18: Studying the impact of somatic mutations of RSK in cancer ............................................................................................... 20
Internship #19: Characterization of new Ras/MAPK pathway components ................................................................................................. 21
Internship #20: Regulation of protein SUMOylation in response to therapeutic agents used in the treatment of leukemia ...................... 22
Internship #21: Cell cycle and DNA damage regulation of the histone deacetylase Hst3 ............................................................................ 23
Internship #22: Analysis of genetic defects in acute myeloid leukemia ....................................................................................................... 24
IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #1: Understanding the molecular controls enabling the initiation
and the completion of mitosis
Research team:
Dr Vincent Archambault
Cell Cycle Mechanisms in Drosophila Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/vincent-archambault/
Project description:
Cell division is essential to life and is controlled by genes conserved between species. The disruption of
molecular mechanisms involved in the cell cycle often leads to cancer. It is therefore important to understand
these mechanisms in order to find new ways to specifically prevent the proliferation of cancer cells.
Cancer cells undergoing mitosis can be blocked by inhibitors specific to mitosis enzymes or the mitotic spindle
(e.g. Taxol). A promising new approach is to inhibit the pathways that allow cells to exit mitosis and return to
interphase.
The entry into mitosis is initiated by the activation of the cyclin B-CDK1 kinase, which is able to phosphorylate
several proteins to trigger chromosome condensation, breakdown of the nuclear envelope, and the
establishment of the spindle microtubules. It was shown that the phosphatase PP2A allows the
dephosphorylation of several substrates of CDK1, the completion of mitosis and the return to interphase. This
phosphatase must be inhibited to allow entry into mitosis, and we recently highlighted some of the molecular
pathway involved. However, several major questions remain unanswered:
1. What are the substrates of PP2A whose regulation is needed to complete mitosis?
2. What is the role of the subcellular localization of various regulators of mitosis?
3. What other genes and proteins regulate the activity of PP2A?
This internship project aims at addressing these questions. To achieve this, the trainee will be working with the
Drosophila fly model, which will allow the combination of powerful genetic tools and techniques related to
molecular biology, biochemistry, proteomics and microscopy.
Techniques used:
Cell culture, molecular biology, real-time imaging, biochemistry, proteomics
Key words:
Cancer, cell cycle, mitosis, kinases, phosphatases, Drosophila
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #2: β-arrestin-dependent regulation of NF-kB signalling by
G protein-coupled receptors
Research team:
Dr Michel Bouvier
Molecular Pharmacology Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/michel-bouvier/
Project description:
Constitutive activation of the transcription factors NF-κB is a common feature of hematologic malignancies,
contributing to their survival and tumor progression, and has been found to reduce the efficacy of many
common cancer therapies, such as chemo- and radiotherapy. NF-κB is therefore being seen as a potential
target for the development of anti-cancer therapies. The activation of NF-κB is controlled by the inhibitory
protein IkB that keeps NF-κB mostly in the cytoplasm. Signal-induced activation of NF-κB is proceeding by
phosphorylation and rapid degradation of IkB by the ubiquitin proteasome pathway. As a consequence of IkB
degradation, NF-κB translocates into the nucleus, where it activates transcription of genes encoding stressresponse enzymes, cell-adhesion molecules, proinflammatory cytokines, and antiapoptotic proteins. Recently,
IkB has been found to interact with β−arrestins (βarr). The βarr are multifunctional molecules that, in addition
to their well-established role in desensitization and endocytosis of G protein-coupled receptors (GPCRs), were
found to bind to various molecules including Mdm2, IkB and IKK. The interactions of βarr with these molecules
modulate phosphorylation, ubiquitination and subcellular distribution of their binding partners. It has been
shown that βarr can modulate tumorigenesis, but the possible impacts of βarr function in tumorigenesis are
just beginning to be appreciated and further studies are needed. The fact that βarr interacts with both Mdm2
(a regulator of p53) and IkB, (a regulator of NF-κB) offers a potential regulatory point that could be involved
the reciprocal regulation between p53 and NF-κB, and contribute to the balance between the tumour
promoter and tumour suppressor activities. Recent studies carried out in our laboratory showed that activation
of many GPCRs including the β1 and β2-adrenergic receptors promote the recruitment of a βarr/IkB/NF-κB to
the receptors that leads to the stabilization of IkB and the inhibition of NF-κB activity.
The present project is aimed at exploring the role of GPCRs and βarr in the control of the IKB/NF-κB complex.
In particular, we will investigate the influence of βarr activation by GPCRs on the ubiquitination and
phosphorylation state of IkB and the dynamics of the IkB/NF-κB complex.
Techniques used:
Human cell culture, BRET, western-blot, immunoprecipitation, gene reporter assays, fluorescence microscopy.
Key words:
Cancer, protein-protein interactions, ubiquitination, G protein-coupled receptors, βarrestin, NF-kB.
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #3: ERM regulation during cell division
Research team:
Dr Sébastien Carréno
Cellular Biology of Mitotis Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/sebastien-carreno/
Project description:
Cell division involves a tight spatiotemporal control of cell shape transformations. Early in mitosis, the cortex
tightens, causing the cell to become round. During anaphase, the poles relax and the equator contracts,
allowing the cell to stretch and eventually split. Then the cellular cortex relaxes to allow cells to re-enter
interphase. Although this series of cell shape transformations were originally described more than 130 years
ago, the underlying mechanisms remain poorly understood. We have established that Ezrin, Radixin, Moesin
(ERM) proteins, control cell morphogenesis during mitosis. During mitosis Drosophila Moesin is activated by
phosphorylation and its localization dictates cell shape transformations. Until early anaphase, dMoe is spread
around the cortex, helping to keep the cell spherical. Subsequently dMoe begins to disappear from the poles
and build up at the equator. We have recently identified a regulatory network that provides a spatiotemporal
framework necessary for mitotic cell morphogenesis. We found that the increase in cortical rigidity that drives
cell shape remodelling at the interphase/mitosis transition involves a Pp1-87B_(phosphatase)/Slik_(kinase)
molecular switch that spatially and timely regulates dMoe phosphorylation and activation.
This research project aims to further understand how dMoe activating networks are controlled. This process
being critical for cancer metastasis, our research will help identify novel cellular targets to design new
therapeutics against cancer.
Techniques used:
Functional genomics, cloning, RNAi, 5D time-lapse microscopy, cell biology
Key words:
Cytokinesis, ERM proteins, actin, cancer
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #4: Regulation of cell division by mitotic kinases: Implications
for cancer development
Research team:
Dr Damien D’Amours
Cellular Mechanisms of Morphogenesis During Mitosis and Cell Motility
Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/damien-damours/
Project description:
This project aims at studying the role of the cell cycle machinery in cell proliferation. Defects in the cell cycle
machinery often cause diseases – such as the Nijmegen breakage syndrome– associated with increased
susceptibility to cancer in humans. These diseases are characterized by defects in the regulation of molecular
signalling pathways responsible for the detection of DNA damage (i.e., checkpoints). Understanding the
molecular defects responsible for this type of disease is an invaluable opportunity to identify the cellular
factors responsible for the development of cancer. This research project integrates several key aspects of cell
biology, such as the regulation of chromatin structure and its connection to cell cycle kinases and signal
transduction pathways. Through this internship, the trainee will be exposed to cutting-edge systems biology
approaches combined with advanced techniques in proteomics, microscopy and genetics.
Techniques used:
Cell biology, molecular biology, microscopy, proteomics, genetics
Key words:
Cell divison, mitosis, kinases
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #5: Generation of Drosophila mutants for candidate
Rab11-GAP proteins
Research team:
Dr Gregory Emery
Vesicular Trafficking and Cell Signalling Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/gregory-emery/
Project description:
During this internship, the student will generate mutants for GAP proteins acting potentially on the small
GTPase Rab11. The student will then test for the effect of the loss-of-function of these proteins on asymmetric
cell division in the peripheral nervous system and during collective cell migration.
Techniques used:
Drosophila genetics, cell biology, molecular biology
Key words:
Small GTPase, Drosophila, cell migration, asymmetric cell division
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #6: SCL and LMO1 oncogenes disrupt thymocyte development
Research team:
Dr Trang Hoang
Hematopoiesis and Leukemia Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/trang-hoang/
Project description:
SCL and LMO oncogenes are frequently activated by chromosomal translocations in T-lineage acute
lymphoblastic leukemia (T-ALL) in children. We have a transgenic mouse model in which the SCL and LMO1
oncogene induce leukemia. Our results indicate that the oncogenic background inhibit differentiation in the
thymus by inhibiting the activity of the transcription factors HEB and E2A, two factors essential to the
development of T lymphocytes. The goal of this project is to determine the molecular basis by which SCL and
LMO1 oncogenes induce leukemia.
Techniques used:
Flow cytometry, cell cycle analysis, quantitative RT-PCR, western-blot, immunofluorescence
Key words:
Oncogene, leukemia, T lymphocyte, transcription factors
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #7: Evaluation of chemical inhibitors of motor proteins in vitro
and in live cells
Research team:
Dr Benjamin Kwok
Chemical Biology of Cell Division Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/benjamin-kwok/
Project description:
The formation of the mitotic spindle, a microtubule-based machine, is required for chromosome segregation
during cell division. Inhibition of spindle assembly blocks cell division and is a viable mean to treat cancer.
Paclitaxel, one of the most successful chemotherapeutics, targets tubulin, which is the building block of
microtubules, and inhibits its polymerization dynamics. However, its success has been limited by the
development of drug resistance in patients. Therefore, alternative strategies are needed to overcome this
hurdle.
Kinesin motor proteins which have the ability to control microtubule organization and polymerization dynamics
provide attractive targets for chemical inhibition. Recently, we have completed two high-throughput screens
to identify small molecule chemical inhibitors for kinesins. From 110,000 compounds that we have screened,
we obtained about more than one hundred candidate hits with different level of selectivity against different
kinesin families. We are in the process of characterizing the lead compounds in vitro using biochemical assays
and in cells using high-resolution microscopy. This internship project is to help determine the effects of these
compounds on enzymatic activity of kinesin motor proteins and on mitotic processes such as spindle assembly
and chromosome segregation.
Techniques used:
Biochemistry, cell culture, microscopy
Key words:
Chemical biology, cancer, microscopy, cell biology, motor proteins
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #8: Characterization of a mitotic motor protein (kif14) that is
linked to cancer progression
Research team:
Dr Benjamin Kwok
Chemical Biology of Cell Division Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/benjamin-kwok/
Project description:
The mitotic kinesin kif14 has been shown to be overexpressed in multiple cancers. Its overexpression correlates
positively with disease progression and poor prognosis. In addition, kif14 is a prime candidate oncogene on
chromosome 1q32, a hot spot of genomic gain found in many of these cancers. Depletion of kif14 in cultured
human cancer cells lead to chromosome segregation defects, cytokinesis failure, and apoptosis. Despite its
importance, kif14 is one of the most understudied kinesins. There are less than 20 articles on kif14 published
to date. The molecular basis of kif14’s role in tumorigenesis and in mitosis remains largely unknown. We have
successfully purified active recombinant kif14 constructs. The goal of the internship project is to characterize
kif14 protein biochemically in vitro and validate our findings in living cells. Results obtained from this study will
be crucial to understand kif14 function and to develop new therapeutic strategies to treat cancer.
Techniques used:
Molecular biology, protein biochemistry, high-resolution microscopy, cell culture
Key words:
Cell division, protein biochemistry, microscopy, cancer, molecular motors
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #9: Characterization of the activity of the tumor suppressor
PAR-4/LKB1 in cell polarity and asymmetric cell division.
Research team:
Dr Jean-Claude Labbé
Cell Division and Differentiation Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/jean-claude-labbe/
Project description:
The goal of this project is to understand the signaling pathways that are regulated by the activity of the tumor
suppressor PAR-4/LKB1, a kinase important for cell polarity. RNAi screens in C. elegans allowed us to identify
regulators of PAR-4/LKB1 activity. The trainee will use specific, imaging-bases assays to define the role of these
regulators in PAR-4/LKB1 function. As mutations in this kinase cause Peutz-Jeghers syndrome, in which patients
develop cancers at high frequency, this work will allow a better understanding of the role of LKBA in human
tumor formation.
Techniques used:
Real-time microscopy, confocal microscopy, RNAi, nematode genetics
Key words:
C. elegans, cellular polarity, tumor suppressor, cancer
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #10: Identification and characterization of virus/host
interactions in modulating innate antiviral responses
Research team:
Dr Daniel Lamarre
Molecular Immunovirology Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/daniel-lamarre/
Project description:
Upon viral infection, recognition of intracellular foreign nucleic acids is made by specific RIG-I-like receptors (RLRs). These
RLRs associate with the mitochondrial antiviral signaling (MAVS) adaptor and lead to the activation of key transcriptional
factors such as IRF3 and NF-κB, induction of type I IFN, and ultimately production of hundreds of ISGs. This antiviral
effector program is a fundamental target for virus-encoded immune suppression, as exemplified by the hepatitis C virus
(HCV) NS3/4A protease that cleaves MAVS to block expression of IFNB1 and IFN-mediated cellular antiviral response. This
viral interference could allow HCV to persist for numerous years in the cytoplasm of hepatocytes, where it is responsible
for chronic liver diseases, such as cirrhosis and hepatocellular carcinoma. The severe adverse effects and limited efficacy
of the actual therapy illustrates the urgent need for the development of new antiviral strategies.
As an obligate parasite, HCV proteins recruit cellular machineries during host infection to ensure efficient viral replication
and evade the host immune system. We propose that an effective way to treat viral infection is to target these virus-host
interactions modulating the host innate antiviral response to either restore immunity upon virus evasion strategies or
prevent excessive inflammatory responses. Using immunoprecipitation (IP) coupled to mass spectrometry (MS) technique,
our group identified 93 host proteins specifically interacting with one of six HCV proteins: Core, NS2, NS3/4A, NS4B, NS5A
or NS5B. The importance of these host proteins for HCV replication was assessed in a previous project. In the second
phase of this project, we used a functional RNA interference (RNAi) screen (5 independent shRNA sequences per gene) to
evaluate the potential modulator role of these 93 potential HCV interactors proteins on the host innate immune response
to viral infection. HEK 293T and A549 cells stably expressing the luciferase gene under the control of the IFNB1 promoter
was used to monitor type I IFN expression in response to Sendai virus (SeV) infection. We selected highly prioritized genes
for which at least two independent shRNAs significantly altered innate antiviral responses. Bioinformatics analyses were
performed using DAVID for Gene ontology (GO) term enrichment and Ingenuity Pathway Analysis (IPA) for host protein
interaction networks.
For selected hits, we will perform WB analysis to confirm the virus-host interaction by co-IP of FLAG tagged HCV proteins,
the knockdown efficiency of selected shRNAs and their effect on antiviral responses and HCV replication. We will
preferentially characterize hit genes for which the knockdown increases innate antiviral responses, while diminishing HCV
replication. Epistasis experiments, relying on the activation of the RLR pathway at different landmarks of the signaling
cascade, will further suggest a potential mechanism of action for these host proteins. Confocal microscopy of Huh7
hepatocytes infected with HCV will also be performed to observe potential co-localization between selected host and viral
proteins.
This research project will increase our knowledge of virus/host interactions and their importance in modulating innate
antiviral responses. Specific inhibitor targeting host proteins characterized in this work could lead to the development of
immune-based therapies to treat viral infections.
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #11: Complementary roles of BAF60A and BAF60B during the
lymphoid vs myeloid differentiation
Research team:
Dr Julie Lessard
Chromatin Structure and Stem Cell Biology Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/julie-lessard/
Project description:
In mammals, BAF chromatin remodelling complexes (Brg/Brm associated factor) are composed of a dozen
subunits assembled around two alternative ATPases, Brg and Brm. The variety of complexes resulting from the
combinatorial assembly of BAF subunits seems to confer functional specificity that allows recognition of
distinct gene targets during cell differentiation. To study the role of the BAF60a BAF60b subunits in adult
hematopoiesis, our lab generated knockout mice for these genes. The deletion of BAF60a in neonatal mice
induces a lack of B- and T-cell precursors in the bone marrow and severe atrophy of the thymus, suggesting an
essential role for BAF60a in lymphopoiesis. Although BAF60b seems dispensable for lymphopoiesis, it is
essential for granulocyte differentiation: the deletion of BAF60b induced severe neutropenia characterized by
an accumulation of granulocyte-macrophage progenitors (GMPs) in the bone marrow and a diminution of the
differentiation to granulocytes. The generation of hematopoietic chimeras showed that this phenotype is cellintrinsic and independent of the microenvironment.
Quantitative PCR (Q-PCR) assays helped us identify several potential targets for BAF60b and BAF60a that could
mediate their function in lymphoid and granulocytic differentiation, respectively. Interestingly, mutations in
several of these genes are associated with lymphopenia and neutropenia syndromes in humans. In the coming
months, we want to assess their importance for BAF60a and BAF60b functions by conducting transcomplementation assays in BAF60a- and BAF60b-deficient cells. In summary, this project should help us to
better understand the complementary role of BAF60a and BAF60b subunits in lymphoid and myeloid
differentiation and potentially develop new treatments for various hematological diseases.
Techniques used:
Flow cytometry, cell culture, retroviral infection, bone marrow transplant
Key words:
Hematopoietic stem cells, BAF complex chromatin
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #12: Quantitative imaging of chromosome dynamics
Research team:
Dr Paul S. Maddox
Mitotic Mechanisms and Chromosome Dynamics Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/paul-maddox/
Project description:
In the Maddox lab, we use state of the art light microscopy techniques to investigate and discover fundamental
aspects of chromosome biology. Specifically we study three topics. 1) how are chromosomes dynamically
remodeled during mitosis 2) what are the mechanisms ensuring epigenome stability through cell division, and
3) how are forces transmitted from the mitotic spindle to segregate chromosomes during cell division. We
have opportunities in each of these subjects, and participants will be exposed to techniques including basic cell
biology, molecular biology, biochemistry, and high resolution imaging. Applicants should be self motivating,
curious, and excited by discovery.
Techniques used:
Cell biology, molecular biology, biochemistry, high resolution imaging
Key words:
Mitosis, chromosome, mitotic spindle, cell division
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #13: Discovering the early events of cell shape change during
physical cell division
Research team:
Dr Amy Shaub Maddox
Cytoskeletal Dynamics and Cell Division Research Unit
Contact information :
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/amy-shaub-maddox/
Project description:
Cell division is essential for life; it is necessary for growth of a fertilized egg into a multicellular organism, for tissue
replenishment, and for wound healing. Failures in cell division are implicated in the development cancer, a prominent and
devastating cause of death in Canada. The final step of cell division is called cytokinesis, when a cell changes shape
dramatically to physically split into two daughter cells. While much of the molecular “parts list” required for cytokinesis is
known, it is poorly understood how these proteins interact to change cell shape – in part because it is challenging to
characterize cell shape in four dimensions (3D + time).
We have recently developed custom image analysis software to describe and measure cell shape and its evolution during
cytokinesis. We can also correlate shape changes with the distributions of fluorescently labeled proteins in order to
understand their roles in cytokinesis. This will allow us to better understand how the cell transforms its shape during
division, and how cytokinesis might be perturbed in cancer cells.
During this internship, the student will become expert at analyzing our collection of 3D time-lapse movies using the new
custom software. The programming has been done in Matlab, but experience with coding is not necessary to operate the
software. The student will then work with lab members to interpret the analysis results, to determine what events are
consistent and which are variable. The student will meet regularly with the lab director and attend weekly group meetings
and other local gatherings. If time permits, the possibility of learning C. elegans embryo dissection, mounting and image
acquisition exists. There is a strong possibility that the work performed during this stage will contribute to a publication. A
possibility of continuing part time into the school year exists.
Techniques used:
Computer-aided image analysis, statistical analysis of data, C. elegans culture, live-cell imaging
Key words:
Cell division, image analysis, bioinformatics, actomyosin contractility
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #14: Production de souches transgéniques de C. elegans à l’aide
de transgènes fluorescents
Research team:
Dr Amy Shaub Maddox
Cytoskeletal Dynamics and Cell Division Research Unit
Contact information :
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/amy-shaub-maddox/
Project description:
Cell division is essential for life; it is necessary for growth of a fertilized egg into a multicellular organism, for tissue
replenishment, and for wound healing. Failures in cell division are implicated in the development cancer, a prominent and
devastating cause of death in Canada. The final step of cell division is called cytokinesis, when a cell changes shape
dramatically to physically split into two daughter cells. While much of the molecular “parts list” required for cytokinesis is
known, it is poorly understood how these proteins interact to change cell shape – in part because it is challenging to
characterize cell shape in four dimensions (3D + time).
We use high resolution live-cell imaging to visualize cytokinesis. To detect a specific cellular protein, we use cells
expressing transgenes encoding our protein of interest tagged with a fluorescent protein. For example, in the images
below, fluorescent markers are expressed for 1) both microtubules and DNA, 2) the contractile cytoskeleton, and 3) the
plasma membrane. The generation of DNA plasmids encoding such transgenes is an essential step towards understanding
cell shape change.
During this internship, the student will become expert at molecular biology. Hands-on experience would be a plus but is
not necessary. The student will work with a lab member to learn our routine methods. With time, the student will become
responsible for prioritizing and troubleshooting the construction and amplification of multiple plasmids. In order to
generate transgenic C. elegans worm strains, completed plasmids will be injected into worms by a senior member of the
lab. The student will meet regularly with the lab director and attend weekly group meetings and other local gatherings. If
time permits, the possibility exists to learn the genetics and screening required to select, identify and maintain C. elegans
transgenic strains. There is a strong possibility that the work performed during this stage will contribute to a publication. A
possibility of continuing part time into the school year exists.
Techniques used:
Molecular Biology, DNA preps, C. elegans genetics and handling
Key words:
Recombinant DNA, cell division, actomyosin contractility
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #15: Mechanisms of action of full antiestrogens in breast
tumor cells
Research team:
Dr Sylvie Mader
Molecular Targeting for Breast Cancer Treatment Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/sylvie-mader/
Project description:
About 2/3 of breast tumors express or overexpress the estrogen receptor and its growth is stimulated by
estrogen. Anti-estrogens are competitive inhibitors of estrogen receptors. There are two classes of antiestrogens, which act by different mechanisms. The goal of this project is to characterize the mechanisms of
action of anti-estrogen such as fulvestrant, a drug used as a second-line therapy for tumors that are resistant to
tamoxifen. Our results indicate that anti-estrogens induce SUMOylation of the receptor and its interaction with
a chromatin remodeling complex. The goal of the project is to characterize the importance of these effects in
the anti-estrogenicity of fulvestrant.
Techniques used:
Cell culture, western-blot, chromatin immunoprecipitation, luciferase assay
Key words:
Breast cancer, antiestrogens, fulvestrant, SUMOylation
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #16: Studying the structure of micro-RNA whose genes are
involved in some cancers
Research team:
Dr François Major
RNA Engineering Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/francois-major/
Project description:
In our laboratory, we want to understand and characterize human gene regulatory networks; link cell content
to phenotypes; and, design and create artificial molecular components to peek and poke cell information and
programmes. We are also developing a suite of programs to analyze, model, predict, and determine RNA 2D
and 3D structure.
During the internship, the student will be examining the structures of micro-RNA whose genes are located in
areas that are under the influence of chromosomal rearrangements or are known to be modified during the
development of cancers. The goal of the project is to identify genetic changes in micro-RNA that could be
related to the etiology of the disease and thus related in a mechanistic way to the progression of cancer.
Techniques used:
Bioinformatics, dynamic programming, constraint programming, predicting RNA structure with programs
developed by our laboratory
Key words:
Micro-RNA, chromosomal rearrangements, bioinformatics, cancer
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #17: Discovering better targets for immunotherapy cancer
Research team:
Dr Claude Perreault
Immunobiology Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/claude-perreault/
Project description:
We have shown that we can cure cancer in mice (e.g. leukemia, melanoma) by injecting CD8 T cells directed
against specific antigens expressed on the surface of cancer cells. Before using this approach in humans, we
must find out why it works with some antigens and not with others. In other words, what are the
characteristics that make an antigen a good target? We postulate that three factors may be involved: i) the
level of antigen expression on cancer cells, ii) the number of T cells that recognize this antigen, iii) the affinity of
the interaction between CD8 and the antigen.
We identified by mass spectrometry eight antigens present on a leukemia cell line (EL4). The eight antigens are
immunogenic, and T lymphocytes immunized against each of them can kill EL4 cells in vitro. However, while the
injection of T cell immunity against four antigens p is capable of removing EL4 cells in vivo, T cells directed
against the four antigens are ineffective. In the coming months, we want to use these eight antigens to
evaluate each of the three factors identified above. We believe that our experimental model will allow us to
identify the essential characteristics of antigens capable of inducing an adequate immune response against
cancer.
Techniques used:
Cell culture, flow cytometry, cell sorting
Key words:
Cancer, T cell, major histocompatibility complex
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #18: Studying the impact of somatic mutations of RSK in cancer
Research team:
Dr Philippe P. Roux
Cellular Signalling and Proteomics Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/philippe-roux/
Project description:
Proteins from the RSK kinase family (p90 ribosomal S6 kinase) are involved in cell proliferation, cell survival and
cell motility. These kinases are normally activated by the Ras/MAPK signaling pathway, which is itself
frequently overactive in cancers, including melanoma, lung and intestine cancer. With the help of highthroughput sequencing of human tumors, several studies have identified somatic mutations in the genes
encoding RSK isoforms (Rsk1-4). The purpose of this project is to generate these mutations experimentally and
to verify their impact on the catalytic activity. With the help of a doctoral student, the student will have to
generate point mutations by site-directed mutagenesis and to verify the integrity of the mutated proteins.
These proteins will then be expressed in human cells and catalytic activity will be measured in vitro and in cells.
This project will help determine whether the somatic mutations that are found in the genes encoding the RSK
family contribute to cancer progression.
Techniques used:
Western blot, transfection, cell culture, mutagenesis, subcloning, kinase assay
Key words:
Kinase, phosphorylation, cancer, somatic mutations
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #19: Characterization of new Ras/MAPK pathway components
Research team:
Dr Marc Therrien
Intracellular Signalling Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/marc-therrien/
Project description:
The Ras/MAPK pathway plays a central role in the control of cell proliferation and its aberrant up-regulation
often leads to cancer. Signaling through the Ras/MAPK pathway depends on a number of core pathway
components that include a module a three kinases known as RAF, MEK and MAPK. We have recently
completed a genome-wide functional (RNAi-based) screen to identify additional factors modulating Ras/MAPK
signaling. Interestingly, this led to the identification of several proteins that appear to control the levels of core
pathway components. In particular, we identified two putative transcription factors that specifically control the
level of mek mRNAs. A position for a summer student is available in the laboratory for validating these findings
and then initiating a characterization of the role these two factors play in controlling MEK levels.
Techniques used:
Molecular biology, biochemistry
Key words:
Ras/MAPK, kinases, transcription factors
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #20: Regulation of protein SUMOylation in response to
therapeutic agents used in the treatment of leukemia
Research team:
Dr Pierre Thibault
Proteomics and Bioanalytical Mass Spectrometry Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/pierre-thibault/
Project description:
This project aims at studying the molecular mechanisms by which therapeutic agents use in the treatment of
acute promyelocytic leukemia (APL) such as arsenite (As2O3) and interferons (IFNs) affect protein SUMOylation.
The small ubiquitin-like modifier (SUMO), is evolutionarily conserved and involved in different cellular
processes including the regulation of intracellular trafficking, DNA repair and replication, cell signaling and
stress responses. Aside from the promyelocytic leukaemia (PML) protein known to be SUMOylated upon
treatment with As2O3 very little is known about the nature of the substrates and their therapeutic significance.
To understand how these agents affect protein SUMOylation we will use metabolic labeling with stable
isotopes to profile changes of this modification in human cells following treatment with therapeutic dose of
As2O3, IFN-α or IFN-γ. This will be performed using quantitative proteomics on a high performance LTQOrbitrap mass spectrometer. These large-scale proteomics analyses will provide valuable insights on the
regulation of protein SUMOylation in response to IFNs and As2O3, ultimately leading to the development of
better therapies for the treatment of leukaemia and other malignancies. Through this internship, the trainee
will be exposed to cutting-edge approaches in molecular biology, protein chemistry, separation sciences,
proteomics, and microscopy.
Techniques used:
Cell culture, microscopy, affinity chromatography, proteomics, mass spectrometry, molecular biology
Key words:
SUMOylation, cell signaling, proteomics
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #21: Cell cycle and DNA damage regulation of the histone
deacetylase Hst3
Research team:
Dr Alain Verreault
Chromosome Biogenesis Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/alain-verreault/
Project description:
Our laboratory discovered a novel and fascinating feature of chromosome physiology. During DNA replication,
chromatin structure needs to be duplicated and this is achieved through the incorporation of newly
synthesized histones into nascent chromatin. We showed that new histone H3 molecules deposited throughout
the genome are acetylated at lysine 56 (H3K56ac). Both H3K56ac and deacetylation play important roles in the
response to a broad spectrum of DNA damaging agents that are frequently used in cancer chemotherapy
(Nature (2005) 436: 294; Curr Biol (2006) 16: 1280; Cell (2008) 134: 244; Mol. Cell. Biol. (2012) 32: 154).
Notably, we found that the enzyme that deacetylates H3K56, known as Hst3, is conserved in many human
pathogens such as Candida and Aspergillus species. Remarkably, inhibition of Hst3 using nicotinamide, a form
of vitamin B3, leads to catastrophic chromosome damage and fungal cell death (Nature Medicine (2010) 16:
775). Thus, inhibitors of Hst3 represent a novel therapeutic approach to treat fungal infections that can be lifethreatening in patients that are immunosuppressed (e.g. patients undergoing cancer chemotherapy or organ
transplant). For these reasons, we are actively studying the mechanism of action and the regulation of the
deacetylase Hst3.
The aim of this project will be to decipher the mechanisms that regulate the activity and the stability of the
Hst3 enzyme during the cell cycle and in response to DNA damaging agents.
Techniques used:
Cell synchronization, immunoblotting, DNA damage sensitivity assays, immunoaffinity purification
Key words:
Histone, Chromatin, Acetylation, Cell Cycle, DNA Damage
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IRIC NEXT GENERATION INTERNSHIP AWARDS 2013
Internship #22: Analysis of genetic defects in acute myeloid leukemia
Research team:
Dr Brian Wilhelm
High-Throughput Genomics Research Unit
Contact information:
 [email protected]
 http://www.iric.ca/en/research/principal-investigators/brian-wilhelm/
Project description:
Recent advances in DNA sequencing technology have made it possible to sequence the RNA from the tumours
of individual AML patients in order to identify all of the mutation present in the disease tissue. This requires
the preparation of patient material before sequencing followed by extensive bioinformatic analysis. To further
dissect the role of individual mutations, the fission yeast model system is also used. The intern’s role in this
project will be to assist in the preparation of material to be sequenced and to carry out specific bioinformatic
tasks. Interns will be required to acquire some fluency with R and Bioconductor software as well as to
demonstrate a clear understanding proper laboratory practices.
Techniques used:
Molecular biology, bioinformatics, analysis of patient data
Key words:
Leukemia, NGS, bioinformatics, mutation discovery, model system
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