MMC Scientific Programs
Stem cell markers for the diagnosis, prognosis and therapeutic stratification of breast, lung and ovarian cancer
Lead scientific contacts
Breast cancer: Salvatore Pece, Fabrizio Bianchi, Stefano Confalonieri, Manuela Vecchi
Lung cancer: Elena Belloni
Ovarian Cancer: Ugo Cavallaro
Stem cells are a rare population of cells necessary for tissue integrity and regeneration in case of injury. Their numbers within any normal tissue are very tightly controlled, but it is now widely accepted that these restraints can malfunction in cancer, leading to an inappropriate expansion of stem cell populations. We discovered that genes expressed in breast stem cells can be used as biomarkers to improve the stratification of breast tumors according to their pathological, molecular and clinical characteristics . However, the original stem cell signature we identified contained too many genes to be of clinical use. Therefore, we are now developing a smaller stem cell profile that still retains the predictive features of the stem cell profile, but that can be analyzed more easily in a clinical setting using immunohistochemistry or quantitative PCR analysis. We are assessing the ability of these biomarkers to classify breast tumors according to aggressiveness, associated metastatic risk and therapeutic response to hormonal and/or chemo-/radio-therapy. Ultimately, we aim to develop a clinical test that will fine-tune the current diagnostic/prognostic ability of clinicians and to improve the clinical management of breast cancer.
Using a similar approach to that pursued for breast , we are now trying to identify and isolate cells with stem properties in non-small cell lung cancer (NSCLC) and in ovarian cancer, for which stem cell identity remains poorly defined. The identification and characterization of the cells that initiate, sustain, and disseminate lung tumors should create new opportunities for the early detection and/or improved treatment for these diseases.
Development of a blood test for the early diagnosis of lung cancer
Lead scientific contacts
The absence of national screening programs for lung cancer, and the lack of symptoms in early stage disease means that the detection of early lung cancer is difficult. The development of clinical tools for the early diagnosis of lung cancer is, therefore, a pressing clinical necessity, particularly for at-risk subjects (smokers or ex-smokers, aged 50 years or more). We have previously identified and validated in independent patient cohorts a circulating miRNA signature (based on 34 miRNA species) that can be used in a blood test to detect early stage lung cancer in asymptomatic patients . In collaboration with theDivision of Thoracic Surgery and the Division of Radiology, we are currently assessing the clinical applicability of this test through its large-scale validation in a prospective multicenter trial launched by the Lung Cancer Early Detection Unit in September 2012 (COSMOS II). The trial has already recruited close to 6,000 of the planned 10,000 patients. If effective, this blood test will be significantly cheaper and easier to apply in national lung cancer screening programs than the only currently available clinical test, which is based on low-dose computer assisted tomography screening.
Understanding breast and ovarian cancer for early diagnosis
Lead scientific contacts
Breast cancer: Fabrizio Bianchi
Ovarian Cancer: Ugo Cavallaro, Fabrizio Bianchi
This scientific program is designed to investigate whether prognostic/diagnostic miRNA profiles exist for breast and ovarian cancers. Based on our experience in developing a blood test to detect early stage lung cancer, we are now collaborating with the Molecular Senology Unit of the Division of Senology, and the Gynaecologic Oncology Unit in serum screening projects, involving breast and ovarian cancer patients, to identify a diagnostic cancer-specific miRNA profile. If we are successful, a small blood sample is all that will be necessary to determine if these miRNAs are present, and this could be sufficient to diagnose breast or ovarian cancers. Our work will lead to the development of a comparatively inexpensive first level diagnostic test for breast and ovarian cancers that can easily be implemented within national screening programs. We have previously shown that more aggressive breast cancers tend to have a larger number of stem cells than less aggressive tumors . Within the context of this program, we are also analyzing the miRNA profile of breast stem cells. If these stem cell-specific miRNAs are also present within the sera of breast cancer patients, they might reflect the proportion of stem cells within a tumor. Though we are still a long way from our endpoint, we hope that testing for the presence of circulating stem-cell miRNAs in blood could form the basis for a prognostic test for breast cancer, which will complement the clinico-pathological parameters that are currently used to assess the aggressiveness of the disease.
New discoveries to feed cancer research
Lead scientific contacts
Breast cancer: Salvatore Pece, Ivan Colaluca, Daniela Tosoni
Lung cancer: Elena Belloni, Fabrizio Bianchi
Ovarian Cancer: Ugo Cavallaro
Our translational medicine program would be short-lived if it were not continually fed by new discoveries in basic research that are improving our understanding of the molecular mechanisms that lie at the root of normal and cancer cell behavior. A number of these projects were born in the laboratory of Pier Paolo Di Fiore, while others are fostered directly within the MMC scientific programs. Amongst these, we are studying the molecular mechanisms that regulate normal stem cell division and we are defining how these are subverted in breast cancer. We are also characterizing critical signaling/endocytic pathways involving Numb/Notch/p53 [12, 13] and factors that cause loss of Numb in cancer, with a view to developing targeted drugs for lung and breast cancer. Several members of the group are involved in high-resolution studies on individual markers that were identified in past genome-wide screening projects, and that potentially have significant clinical relevance as prognostic or therapeutic targets. Our ovarian cancer group is generating molecular tools for the identification and characterization of ovarian cancer stem cells, whose precise identity still remains uncertain today. As well as these classical approaches to research, our group is using cutting edge next-generation sequencing technology to define the genetic determinants of lung cancer in smokers and in non-smokers, and to analyze the clonal origin of breast and lung cancers and their metastases.
MMC Support Infrastructures
Lead scientist: Giuseppina Bonizzi
Technicians (Tissue Bank): Marialucia Longo, Giulio Taliento, Andrea Uggetti
Technician (Lab Support): Alessandro Di Nasta
Research Nurse: Anna Carmagnola
Data entry/Data manager: Maria Capra, Fulvia Fusar
The Tissue Bank (IEO Biobank and Biomolecular Resource Infrastructure - IBBRI) collects, catalogues and stores biological samples (surgically excised tissue samples non-essential for diagnosis, plasma/serum, total blood, DNA and RNA) from patients who have provided informed consent for the use of their tissues for research purposes. We have established a direct pipeline with the operating theatres for the collection of tissue samples and these are stored according to specific protocols and standard operating procedures. We also store any primary cell cultures, stem cell preparations, and xenotransplanted tumors derived from collected samples. All biobanked samples are managed and tracked through a software package that is fully integrated with the hospital medical records database, pathology database and central registry of patient demographic information. This ensures that each sample is linked to a full complement of anonymous or anonymized (according to patient choice) patient information (accessible solely by authorized Biobank personnel). The high quality biospecimens we collect are used for biomarker and drug discovery experiments, both for basic research and for MMC translational research projects.
Molecular Pathology Laboratory
Lead scientist: Stefano Confalonieri
Pathologist: Giovanni Bertalot
Postdoctoral Fellow: Elena Miranda
Technician: Giovanna Jodice, Chiara Luise, Valentina Mattei
The IEO Molecular Pathology Laboratory works to identify novel putative cancer targets for drug discovery, diagnostic and/or prognostic applications through high-throughput Tissue Microarrays (TMAs) and in situ detection methods (in situ hybridization and immunohistochemistry). TMA technology allows gene expression analysis on thousands of patient tissues simultaneously, and is a powerful tool for determining the relevance of basic research findings to cancer.
We collaborate closely with the Pathology Department at IEO, who maintain tissue archives dating back to 1994, when IEO was established. These are an invaluable source of material for the construction and continuous replenishment of our TMAs. Thanks to data stored by the IEO Tumor Registry, all arrayed tissues are linked to complete clinico-pathological and follow-up information. The Molecular Pathology Laboratory can therefore link the expression patterns of genes of interest to clinical evaluation and outcome, a necessary step for the validation of new diagnostic and prognostic markers.
Primary Epithelial and Stem Cell Culture /Xenotransplant Laboratory
Lead scientist: Tamara Tanos
Staff scientist (xenotransplants): Daniela Tosoni
Technicians (cell culture): Rossana Bettolini, Ivan Cuccovillo, Donatella Genovese
Technicians (xenotransplants): Barbara Giulini , Stefania Amodio
The Primary Epithelial and Stem Cell Culture Laboratory derives bulk primary epithelial cells and stem cells from human biopsy specimens. Primary-derived cells are used for the establishment of either in vitro or in vivo (xenotransplant) model-systems that retain many features of their parental tissues. These are a valuable tool for studying the molecular features of naturally occurring human cancers. We have developed efficient protocols for the processing of tissue biopsy specimens, and for the preparation of bulk primary epithelial cell and stem cell cultures from breast , lung, ovary and prostate tissues, and for their xenotransplantation in immunocompromised murine hosts. Samples arrive to our laboratory via a continuous pipeline from the operating theatres, coordinated by the Tissue Bank.
Clinical Biomarkers Laboratory
Lead scientist: Manuela Vecchi
Scientist: Flavia Troglio
Technicians: Francesca de Santis, Luca Napolitano, Stefania Pirroni
The Clinical Biomarkers Laboratory performs screening, pre-clinical validation, and optimization of biomarker candidates, as well as the design, execution and technological development of cancer diagnostic assays to aid the translation of candidates markers into the clinical setting. In this role, we also support collaborative study programs between biotech companies, researchers and clinicians for the co-development of biomarkers and novel therapies. This facility is also responsible for the optimization of protocols for the extraction of nucleic acids (including total RNA, DNA and miRNAs) from human tissues, such as blood, plasma, fresh tissue biopsies, paraffin-embedded tissue specimens. We also set up and implement novel technological platforms for the group. Finally, we select the appropriate patient cohorts and tissue samples for prospective and retrospective studies in collaboration with the Genomics and Bioinformatics Laboratory.
Genomics and Bioinformatics Laboratory
Lead scientist: Fabrizio Bianchi
Postdoc: Francesca Montani
PhD students: Eleonora Lusito, Valentina Melocchi, Simona Monterisi
Fellows: Elisa Dama (co-supervised by Eng. Patrick Maisonneuve, Epidemiology and Biostatiscs Division, IEO), Fabio Dezi
Technicians: Rosemary Carletti
The Genomics and Bioinformatics Laboratory draws together the fields of genomics, biostatistics and applied bioinformatics. We apply ‘omics strategies’ to identify novel cancer biomarkers and potential therapeutic targets and use computational biology and biostatistical approaches to prioritize cancer biomarkers. We are primarily involved in the screening for circulating miRNA biomarkers for the early diagnosis of lung, breast and ovarian cancer patients, and in the transcriptome analysis of primary tumors and cancer stem cells.
One of our key roles is to provide support to other MMC units. We are responsible for: i) coordinating, centralizing and managing clinical information associated with biological samples collected by the Tissue Bank; ii) designing and analyzing studies involving the screening, pre-clinical validation, and optimization of biomarker candidates, identified by the Clinical Biomarkers Laboratory.