Dr Elena Rainero
Room: B2-04 Florey building
Brief career history
Extracellular matrix internalisation in breast cancer invasion
The extracellular matrix (ECM) is a complex network of secreted proteins which not only provides tissue support, but is also involved in the control of several cell functions, including migration and oncogenic transformation. The tumour microenvironment has a pivotal role in modulating cancer initiation, progression and metastasis, while cancer cells in turn modify the composition and properties of the ECM, demonstrating a bi-directional interaction between tumour and stroma. Our research addresses cell-ECM interaction from a novel and exciting angle, investigating how the internalisation of ECM components control breast cancer cell invasion.
Using a combination of different approaches, we aim at characterising how ECM internalisation controls cancer cell migration and invasion in 2 dimensions (2D), 3 dimensions (3D) and in vivo systems.
Extracellular matrix internalisation in epithelial remodelling
Most of our organs are built from epithelial cells. They are supported extracellularly by a complex network of secreted proteins collectively known as extracellular matrix (ECM). The ECM is commonly viewed as “dead space” that provides a static scaffold for organ shape and acts as an obstacle for migrating cells. However, new data strengthen the idea that there is a dynamic interplay between cells and the ECM, which actively orchestrates how cells and organs acquire their shape. Therefore, a better understanding of how cells dynamically remodel their ECM environment and how the cell-ECM interplay contributes to epithelial remodelling will shed new light on fundamental mechanisms underlying the shaping of organs.
The mammary gland has a tree-like structure, whereby a single monolayer of epithelial cells surrounds the lumen of the ducts. The formation of new branches, or branching morphogenesis, is controlled by hormones and growth factors, including Hepatocyte Growth Factor (HGF). Moreover, the extracellular matrix (ECM) has been shown to have a key role in this process. Our data indicate that, upon HGF stimulation of normal mammary epithelial cells (MECs), tubule formation is associated with ECM remodelling and uptake, suggesting that ECM endocytosis might contribute to branching morphogenesis.
We aim at characterising the endocytic pathway(s) responsible for ECM uptake, focusing on the integrin family of ECM receptors, and assessing the contribution of ECM uptake to branching morphogenesis in 3D environments
Undergraduate and postgraduate taught modules
Postgraduate PhD studentship
1. Dissecting Extracellular Matrix Internalisation Mechanisms using Functional Genomics
Location: Department of Biomedical Science, The University of Sheffield / Institute of Molecular and Cell Biology, A*STAR institute, Singapore
This is a 4-year funded PhD project, with 2 years in Sheffield and 2 years in Singapore.
The extracellular matrix (ECM) is a complex network of secreted glycoproteins providing tissue support and controls a variety of cell functions, including tumour growth. Reports dating back to the 1990s have documented the internalisation of ECM components, including collagen and laminin. More recently, ECM endocytosis has been linked to increased matrix degradation by cancer cells. Consistent with this observation, previous work by the Rainero group demonstrated that ECM internalisation is required for cancer cell migration and nutrient signalling, suggesting the intriguing hypothesis that ECM uptake might represent a nutrient source for invasive cancer cells. These observations point to the machinery controlling ECM endocytosis as a novel target for the development of pharmacological intervention to limit cancer spreading.
Preliminary data from the Rainero lab show that ECM internalisation is strongly increased in invasive breast cancer cells, compared to normal mammary epithelial cells and non-invasive breast cancer cells. Cells interact with the ECM through plasma membrane receptors, which has been shown to promote the internalisation of their ECM component ligands. The molecular mechanisms controlling this are poorly defined. To faithfully recapitulate the architecture and composition of the in vivo ECM, cell-derived matrices (CDMs) will be used. These are fibrillar 3D matrices generated by fibroblasts and the tools to study their internalisation have been recently developed in the Rainero lab.
This project will characterise the endocytic mechanisms controlling ECM internalisation using a functional genomic approach and test whether the GALA pathway affects ECM internalisation. This will be achieved using the RNA interference (RNAi) screening technology at the genomic scale, which will allow the identification and accurate quantification of novel key players in this process. The RNAi screen will be performed at the Institute of Molecular and Cell Biology RNAi screening facility (which is embedded in the Bard lab), using the human genome siRNA library and the invasive human breast cancer cell line MDA-MB-231. We will use bioinformatics to construct regulatory networks and subnetworks, to provide a genetic overview of the endocytic pathways controlling ECM internalisation by cancer cells. Based on this analysis, key pathway(s) will be identified and a list of hits will be selected and validated using cells extracted from mouse primary breast tumours. Finally, the contribution of the identified regulators of ECM uptake and the GALA pathway in controlling breast cancer cell proliferation and migration will be investigated, using imaging-based proliferation assays, live cell time-lapse microscopy and 3D invasion assays.
These studies will provide a comprehensive and complementary set of molecular cell biology and genetic approaches, coupled with advanced bioinformatics and imaging techniques to elucidate the molecular mechanisms controlling ECM internalisation. The data generated will provide novel insight into the contribution of regulators of ECM internalisation in the control breast cancer cell proliferation and migration.
Whilst in Sheffield, students receive fees (£4,260 in 2018/19) and an RCUK rate stipend (£14,553 in 2018/19).
Whilst in Singapore, students receive the following:
2. Investigating the role of the protease ADAMTS5 in ovarian cancer
Funding status: Competition funded project European/UK students only
This project is eligible for a department scholarship. These scholarships are awarded on a competitive basis – find out more on our funding webpage.
This project is also open to self funded students
This project focuses on the interaction between cancer cells and the tumour microenvironment, analysing the role of the extracellular matrix (ECM) modifying enzyme ADAMTS5 in ovarian cancer cell invasive migration. In particular, we will dissect how ADAMTS5 protein is upregulated in ovarian cancer cells and the molecular mechanisms through which the cleavage of the ADAMTS5 substrate versican promotes cancer cell invasion. Ovarian cancer is the most lethal gynaecological malignancy.
Mostly because of late stage diagnosis, the 5-year survival rate is <30%. The major cause of death is associated with the presence of therapy-resistant metastasis. Given the fact that >70% of ovarian cancers are diagnosed at late stage, it is essential to understand what the molecular mechanisms controlling metastasis formation are in order to develop novel strategies for the maintenance of this deadly disease. The tumour microenvironment, including the ECM, has a pivotal role in modulating cancer initiation, progression and metastasis. In particular, Versican (VCAN), a proteoglycan of the lectican family, has been shown to be upregulated in brain tumours, melanomas, osteosarcomas, lymphomas, breast, prostate, colon, lung, pancreatic, endometrial, oral and ovarian cancers and high VCAN expression correlates with reduced overall survival in ovarian cancer patients. VCAN is cleaved by a family of zinc-dependent metalloproteases, A Disintegrin And Metalloproteinase Domain with TromboSpondin type I module (ADAMTS), composed of 19 members of secreted proteases, including ADAMTS5. ADAMTS5 has been shown to increase cell invasive potential in glioblastoma, lung cancer, colon cancer and laryngeal cancer cells. Vesicular trafficking is essential for the polarised distribution of transmembrane receptors and secreted molecules and it is controlled by Rabs, small GTPases of the Ras family.
It is not surprising that alterations in Rab expression and function have been associated with cancer. In particular, epithelial specific Rab25, a member of the Rab11 family, has been shown to promote ovarian cancer cell migration and invasion. Consistent with this, high Rab25 expression correlates with poor prognosis in ovarian cancer. Preliminary data from the lab indicate that Rab25 induces ADAMTS5 expression, and the catalytic activity of ADAMTS5 is required for Rab25-induced cancer cell migration and invasion through 3D environments.
Therefore, we hypothesise that Rab25-expressing ovarian cancer cells upregulate ADAMTS5 expression when in contact with fibroblast-generated ECM. This in turn stimulates their migratory and invasive ability, eventually promoting metastasis formation. This project specifically aims at answering the following questions:
Despite the incidence and mortality of ovarian cancer, the mechanisms controlling metastasis are not fully understood. The outcome of this project will deepen our understanding of the interplay between cancer cells and the ECM, highlighting whether ADAMTS5 can represent a novel target for the development of new ovarian cancer therapies.
Keywords: Cancer / Oncology, Cell Biology / Development
For informal enquiries about the projects or application process, please feel free to contact me.
To find out more about this project and how to apply see our PhD opportunities page:
- Rainero E (2018) Extracellular matrix internalization links nutrient signalling to invasive migration. International Journal of Experimental Pathology. View this article in WRRO
- Dornier E, Rabas N, Mitchell L, Novo D, Dhayade S, Marco S, Mackay G, Sumpton D, Pallares M, Nixon C, Blyth K, Macpherson I, Rainero E & Norman JC (2017) Glutaminolysis drives membrane trafficking to promote invasiveness of breast cancer cells. Nature Communications, 8. View this article in WRRO
- Rainero E (2016) Extracellular matrix endocytosis in controlling matrix turnover and beyond: emerging roles in cancer. Biochemical Society Transactions, 44(5), 1347-1354. View this article in WRRO
- Rainero E, Howe J, Caswell P, Jamieson N, Anderson K, Critchley D, Machesky L & Norman J (2015) Ligand-Occupied Integrin Internalization Links Nutrient Signaling to Invasive Migration. Cell Reports, 10(3), 398-413. View this article in WRRO
- Christoforides C, Rainero E, Brown K, Norman J & Toker A (2012) PKD Controls αvβ3 Integrin Recycling and Tumor Cell Invasive Migration through Its Substrate Rabaptin-5. Developmental Cell, 23(3), 560-572.
- Rainero E, Caswell PT, Muller PAJ, Grindlay J, McCaffrey MW, Zhang Q, Wakelam MJO, Vousden KH, Graziani A & Norman JC (2012) Diacylglycerol kinase α controls RCP-dependent integrin trafficking to promote invasive migration. The Journal of Cell Biology, 196(2), 277-295. View this article in WRRO