Crosstalk of cell-cell and cell-matrix signaling in collective migration in development and carcinogenesis
Project Manager: Dr. Mirjam Zegers
Interactions of cells with the underlying matrix or with neighboring cells profoundly affect cellular morphology. My research interests center on the molecular mechanisms underlying the regulation of epithelial morphogenesis by cell-cell and cell-matrix adhesion. Cadherin-mediated cell-cell is critical for embryogenesis, tissue formation and regeneration, but also has an important regulatory role in pathogenic events like cancer. We currently focus on the role of different cadherin family members in collective invasion and invasive growth of developmental branching morphogenesis. We hypothesize that these developmental processes are recapitulated in collective cancer invasion.
Serial Killers in the fight against cancer
Project manager : Prof. dr. Peter Friedl
Researchers: Dr.Cindy Dieteren, Esther Wagena BSc.
This project is funded by the Dutch Cancer Foundation (KWF).
Within our immune system cytotoxix T-lymphocytes are specialized to kill infected cell and cancer cells (target cells). In the effector phase CTL’s find their specific target cell and with its ability to use the deadly lytic granules, the target cell is being killed while the surrounding healthy cells stay unaffected. For treatment of cancer via immunotherapy CTL’s are very interesting and promising. Although its ability to kill a tumor cell, it seems somehow not possible to stop the tumor growth using CTL immune therapy. In this project we aim to gain deeper insight in the effector phase of an anti-tumor immune response in the tumor micro environment, both in vitro as in vivo. Techniques are cell culturing, immunology, mouse models, microscopic imaging.
European Network for Cell Imaging and Tracking Expertise (ENCITE)
Project manager : Prof. dr. Peter Friedl
Researchers: Bettina Weigelin MSc. Ph.D. student, Dr.Gert-Jan Bakker
ENCITE is co-funded by the European Commission under the 7th Framework Programme. The European Network for Cell Imaging and Tracking Expertise belongs to a large scale integrating project of a consortium of 22 partners. The main target is to develop relevant Imaging technologies to better understand how cell-therapy works. The tools and methologies developed in this project will be validated in 5 key disease areas; Neurological, Cardiovascular, Musculoskeletal, Diabetes and Cancer. Within our group the main focus is cancer, we will develop novel reporter genes and generate reporter mice for in vivo imaging.
(we will generate dual color mice which can be used for study cell proliferation, migration, cell trafficking, tissue repair and morphogenesis. Furthermore the dual color approach will be used to study the host response against human tumor xenografts and immune response against transplanted melanoma.)
Furthermore we are involved in the Pre-clinical validation in T-cell therapy and Monitoring dynamics of tumor and tumor micro environment
T3-net, Tissue Transmigration Training Network.
Project manager : Prof. dr. Peter Friedl
Researcher: Olga Ilina MSc. Ph.D. student, Marina Vortmeyer
MSc. Ph.D. student
This project is an initial training network funded by the EU people program (formally known as Marie Curie). It ables us to educate a Ph.D .student in cell tracking. In this project the role of lytic protutions in 3D tissue invasion is investigated with advanced Imaging techniques, including bright field time lapse microscopy, confocal reflexion and ( relatively collagen specific) two-photon excited second harmonic generation microscopy.
Healthy Ageing, Pieken in de Delta Oost Nederland.
Project manager : Prof. dr. Peter Friedl
Researchers: Dr. Jack Fransen, Dr. Pavlo Grytsenco, Huib Croes BSc.
This is an unique consortium between the industry and academic centres both located in the East of the Netherlands. The project is partly funded by Ministry of Economic affairs and the Province of Gelderland.
Everybody wished to age in a health way, unfortunately diseases like cancer, rheumatoid arthritis, diabetics, strokes, Parkinson and Alzheimer diseases are responsible for less quality of life and even responsible for a premature death. The main target in this research project is to develop identification of new drugs and functional foods, regarding Ageing related diseases, in a more efficient and cheaper way. In order to achieve this our group will help to develop human cellular models and use this models for effectivety measurements of identified chemical and food compounds.
Mechanisms of integrin-independent amoeboid modes of cell migration – role of the glycocalyx and physical mechanisms
Project manager:
Prof. dr. Peter Friedl
Researcher:
The migration of cells through the extracellular matrix (ECM) of tissues and organs is a fundamental process in morphogenesis, tissue repair and, in its deregulated form, cancer invasion and metastasis. Adhesion molecules of the integrin family are the major receptors that provide adhesion towards structural proteins of tissues, including fibrillar collagens, laminins, and fibronectin. Due to the dominant contribution of integrins to mechanotransduction and migration in most experimental models, additional low affinity interactions that contribute to cell movement through three-dimensional (3D) ECM remain poorly defined. Aim of this project is to show that integrins are dispensable for substrate binding and migration due to newly emergent amoeboid migration modes of low force, in which the glycocalyx and physical mechanisms generate distinct molecular and physical cell-matrix contacts and migration programs.
Localization of heparan sulfate (HS) at interaction sites with collagen fibres (black arrowheads) on MV3 control cell during migration. HS is accumulated in anterior ruffles and filopodia (inset, white arrowheads) and the trailing edge (gray arrowheads). Asterisk, deposited material from the cell rear, confirming the migratory state. Bars, 10 µm (B, top; F), 100 nm (b, bottom).
Schematic view of different cell surface glycan-classes of the surface glycocalyx on cells.
T cell – tumor cell interactions and serial killing: Impact of the tumor microenvironment
Project manager:
Prof. dr. Peter Friedl
Researcher: Bettina Weigelin MSc. Ph.D. student
Immunological control of progressive tumours requires not only activation and expansion of tumour specific cytotoxic T-lymphocytes (CTL), but also an efficient effector phase. This includes migration of CTL in the tumour, serial conjugation and killing of target cells. Within this project we established a real-time 3D collagen model of CTL function. This allows the observation of active migration, interaction, dissociation and serial killing of single cells. Aims of the continuing project will be: (1) to develop the present in vitro model further by using multicellular target-cell spheroids and improved real-time reconstruction of apoptosis. (2) to describe serial killing in established, antigen-specific tumour models (E.G7/EL4 and B16 melanoma) in vitro and to modulate the system by changing environmental conditions (addition of chemokines, TGF-ß, regulatory T-cells (Treg). (3) finally to validate the results in vivo, using multiphoton microscopy to reconstruct adaptively transferred CTL in orthotopic transplanted “dual-colour” tumours.
Mechanisms and consequences of collective invasion
Project manager :
Prof. dr. Peter Friedl
Researchers: Anna Haeger MSc. Ph.D. student, Olga Ilina MSc. Ph.D. student and Antoine Khalil MSc. Ph.D. student
Collective invasion, in addition to single cell dissemination, has been identified as prevalent invasion mode in many cancer types.
In vitro and in vivo studies strongly suggest, that cancer cells are able to generate functional cell-cell junctions and penetrate tissue as multicellular strands or masses in a coordinated manner and, as outcome, ultimately lead to tissue destruction. Collectivity may further convey altered biology, including survival-promoting signals and altered therapy response.
This project thus aims to identify the in vivo mechanisms and consequences of collective cell invasion for the surrounding healthy tissue, cancer metastasis and therapeutic efficacy. This includes mechanisms of cell-cell adhesion and polarity, the role of fibroblasts in guiding cancer invasion, and how cell-cell communication controls tumor cell survival in melanoma, skin and breast cancer cells.
With focus on cadherins, immunoglobulin family members and connexins as well as cross-talk to the tumor stroma, using both, in vitro and in vivo models, molecular interference as well as advanced microscopy, homo- and heterotypic cell-cell adhesion and communication pathways are addressed. By applying blocking monoclonal antibodies, small molecule inhibitors and RNA interference, the role of each pathway on cell-cell cohesion, transitions to other migration modes and the implications for in vivo metastasis will be identified.
Models and mechanisms of Glioma cell invasion
Project manager :
Prof. dr. Peter Friedl
Researcher: Dr. Pavlo Grytsenko
The high lethality of glioma patients is mainly due to diffuse invasiveness of glioma cells into the brain parenchyma. Within the brain glioma cells migrate along myelinated fibers and the basement membranes of blood vessels that serve as guiding structures for glioma dissemination. The brain parenchyma has a unique composition of cell processes, vessels, and hyaluronic acid as the main component of interstitial matrix.
The aim is to design in vitro assays that recapitulate such brain-like matrix for functional live-cell studies. Using these tools, adhesion and cell-cell communication mechanisms of glioma invasion will be studied, which focus at chemokines, integrins and the cadherin/catenin axis.
Figure: Migration of U-251 glioma cells from a spheroid