Grant Winners

Spring 2014 Research Grant Recipients Talk About Their Research


Elena Di Gennaro, PhD
Staff Investigator
National Cancer Institute, Naples, Italy

Abstract Title:
Evaluation by Multicolor Flow Cytometry of Circulating Endothelial Cells as Prognostic and Predictive Biomarkers in Cancer Patients


BD: What is your educational background?

Elena Di Gennaro: I received my undergraduate degree in molecular biology, plus a special degree in clinical biochemistry, from the University of Naples, Italy, where I remained for my PhD in cellular biochemistry. I was a predoctoral and postdoctoral fellow in the Experimental Pharmacology Unit, Experimental Oncology Department, at Italy's National Cancer Institute (Fondazione G. Pascale), also in Naples. For the last six years I have been a staff investigator within the same department and institute, under the mentorship of Prof. Alfredo Budillon.

BD: How and when did you become interested in science?

Elena Di Gennaro: I do not know exactly, but a crucial moment was when I received a small microscope as a gift for my ninth birthday. As I progressed through school I developed an interest in cancer research.

BD: How did you become interested in your broad field of study?

Elena Di Gennaro: My thesis adviser was an immunologist who allowed me to take courses in immunology, which is why I'm now interested in that field. Our laboratory focuses principally on preclinical and clinical development of novel combinatorial therapeutics for human cancer based on the rational use of targeted and conventional treatments, with the aim of improving efficacy while reducing toxicity. Our studies cover the mechanistic basis of molecular targeted drugs in preclinical models, developing novel pharmacologic associations between biological and conventional anti-cancer agents, identifying new therapeutic targets, and using advanced technology platforms to identify new molecular diagnostic, prognostic, and predictive markers of therapeutic response in preclinical models and from biopsies and bodily fluids of cancer patients. This approach employs proteomics and pharmacogenomics, and determination of circulating endothelial cells, their progenitors, and other circulating factors.

BD: Describe the project for which you received the BD grant.

Elena Di Gennaro: We hypothesize that circulating endothelial cell (CEC) count could represent an important prognostic and predictive biomarker in colorectal and ovarian cancer patients. Evaluating levels—both baseline and during treatment—of CECs could help to select the patients most likely to benefit from anti-angiogenic therapies, and/or to identify possible mechanisms of resistance. These hypotheses could be validated only through a standardized method of analysis and through the definition of a reference range of counts in healthy individuals.

BD: What are the long- and short-term scientific goals of this project?

Elena Di Gennaro: We expect to achieve several goals during this project in collaboration with other Italian research groups. First on the agenda is to standardize how we identify and count CECs in the peripheral blood of healthy donors to define a reference range of CEC counts, and compare those with the basal levels measured in naïve or untreated colorectal and ovarian cancer patients. We will also determine whether the viability of CECs varies within 24 hours after collection and storage at 4°C or after separation of peripheral blood mononucleated cells and storage of cells at -80°C. Next we plan to evaluate correlations between CEC counts and stage of disease, phenotypic characteristics of the tumor, and patient outcomes. In the long term we hope to define the prognostic and predictive value of CEC count on the clinical efficacy of anticancer treatment, including anti-angiogenesis agents.

BD: What are the implications of your project for human health?

Elena Di Gennaro: Colorectal and ovarian cancers are among the most common malignancies. Despite the improvement in diagnosis and treatment, they remain two leading causes of cancer deaths. There is a serious, unmet need for new diagnostic, prognostic, and/or predictive biomarkers that could offer opportunities for early diagnosis or for identifying patients who could most likely benefit from a specific treatment or, conversely, who might avoid unnecessary, ineffective, deleterious, and costly therapies. In both colorectal and ovarian cancers, treatment with bevacizumab plus chemotherapy improves clinical outcome significantly. However, at the moment there is no validated predictive biomarker for the efficacy of bevacizumab, or for that matter any of the anti-angiogenic oncology drugs. We hope that our study validates the basis for using CEC counts as new potential diagnostic, prognostic, and/or predictive cancer biomarkers.

BD: Which BD reagents do you plan to use, and for what purposes?

Elena Di Gennaro: Since we will be using flow cytometry, we expect to acquire several BD reagents and analysis tools, including BD™ CompBead particles and reagents appropriate for the BD FACSCanto™ II system. For staining we will use a lyophilized cocktail of reagents from BD Biosciences, defined on the basis of a panel previously optimized that includes CD146 PE, CD34 PE-Cy™7, CD309 APC, CD45 APC-H7, and 7-AAD, plus the following antibodies: CD31 V450 and/or CD133 biotinilated + streptavidin V500.


David Escors, PhD
Principal Investigator
Navarrabiomed-FMS, Spain

Abstract Title:
Identification of Novel Therapeutic Targets in Breast Cancer-Specific Myeloid-Derived Suppressor Cells


BD: What is your educational background?

David Escors: I studied biology at the University of Navarre, in Spain, and at the University of Buckingham, England, receiving my BSc degree in 1997. I earned my PhD in molecular virology from the Autonomous University of Madrid in 2002, where I remained for three years for my first post-doctoral fellowship. I then moved to University College, London, as a Marie Curie Fellow, joining the research group of Prof. Mary Collins. I subsequently formed my own independent research group at University College in 2008.

BD: How and when did you become interested in science?

David Escors: I wanted to be a scientist from as far back as I can remember. For me it was not a question of "becoming interested" in science, because I was always interested. While my schoolmates were interested in cars and that sort of thing, I knew all the dinosaurs' names, and was learning about pulsars and black holes. So it was natural for me to obtain a science degree and go into basic research.

BD: How did you become interested in your broad field of study?

David Escors: Since 2008, my field of interest has been immunology and how to modify immune responses by gene therapy using lentiviral vectors. But I was interested in immune responses even before I started my university degree. AIDS appeared when I was young, in the early 1980s, and that ignited my interest in immunology and virology. I still remember the TV news with a new strange disease that was rapidly propagating, the excitement of the discovery of the HIV virus and its life cycle. That's probably why I chose viruses as vehicles to modify cells, and then to manipulate immunity. Everyone in the 80s and early 90s was looking for a cure for AIDS, but it never came. When I started my PhD, so many of my colleagues were researching AIDS and lentiviruses that I didn't want to be just another face in the crowd. So I focused my PhD work on coronaviruses. When I eventually earned my doctorate, I began researching HIV-1 lentivirus vectors for gene therapy of the immune system.

BD: Describe the project for which you received the BD grant.

David Escors: A few years ago, researchers noticed that tumors exerted a strong immunosuppressive effect on patients, and a new cell type was described as being responsible for this effect. This cell type was named "myeloid-derived suppressor cell," or MDSC. Research on MDSCs is quite challenging, because they have to be purified from tumors induced in a large number of mice. For example, twenty cancerous mice yield only about three million tumor-infiltrating MDSCs.

That makes MDSC research and drug discovery extremely difficult. So I thought it would be helpful to culture tumor cells as a more-or-less endless supply of MDSCs. Thanks to the work of my PhD students, Noemi Perez-Janices, Therese Liechtenstein, and Ines Dufait, and the contribution of several others, we were able to simulate the tumor environment in vitro. Through this approach we can generate between fifty and seventy million MDSCs in five days from bone marrow obtained from a single mouse, without having to induce cancer in the animal. This invaluable experimental system reduces the number of mice used in research. In the current project, we would like to produce breast cancer-specific MDSCs and analyze them to identify novel targets susceptible to therapeutic intervention.

BD: What are the long- and short-term scientific goals of this project?

David Escors: The short-term scientific goals of this project are to generate breast-cancer MDSCs in vitro, and compare their protein and gene expression profiles with MDSCs that are not cancer-specific. We will also compare them with other immunostimulatory myeloid cells such as dendritic cells. These comparisons will provide us with a molecular map highlighting the intracellular pathways that are activated/deactivated in cancer-specific MDSCs. Long term, we would like to target these cancer-specific pathways identified in breast-cancer MDSCs by interfering with those mechanisms using gene therapy and chemotherapy. This information will be especially useful for developing novel cancer treatments.

BD: What are the implications of your project for human health?

David Escors: We hope to discover novel pathways induced by cancer that are susceptible to therapeutic intervention. The large numbers of cancer-specific MDSCs we can now access will allow high-throughput testing of multiple treatments and chemotherapeutic drugs on these cells. This will allow the discovery of new treatments which specifically eliminate MDSCs or counteract their immunosuppressive activities. Once these cells are removed, cancer can be attacked by the immune system wherever it is found in the organism.

BD: Which BD reagents do you plan to use, and for what purposes?

David Escors: We will use a range of reagents, including antibodies specific for molecules expressed by myeloid cells, such as lineage markers, activation and antigen presentation molecules, and others. As we discover new differentially expressed markers, we will require specific antibodies for their detection and study. In addition, we will employ recombinant proteins to drive myeloid cell proliferation in vitro. This BD award has meant a lot to my group. Because of this grant we can analyze cells using techniques that were previously inaccessible to us.


Thomas Huneck Haupt, MD
Graduate Student
Hvidovre Hospital and University of Copenhagen

Abstract Title:
The Significance of Regulatory T Cells in Crohn's Disease


BD: What is your educational background?

Thomas Haupt: I did my undergraduate studies and received my MD from the University of Copenhagen. I am currently a PhD student at Hvidovre Hospital, working with the institution's research director, Prof. Ove Andersen, as my main supervisor.

BD: How and when did you become interested in science?

Thomas Haupt: I have always been extremely curious, but in my teens I became interested in the humanities. I read several books about famous philosophers like Rene Descartes and David Hume. Their ideas fascinated me, but I was even more intrigued by our human ability and desire to understand everything we see. When I began medical school, I remember imagining—naively—that everything important in medicine had already been discovered. My interest in science was stimulated by the discovery that we actually know very little.

BD: How did you become interested in your broad field of study?

Thomas Haupt: It was during my basic training in immunology. I had a very inspirational teacher, and I am fortunate enough that he is my supervisor today. In my opinion, the immune system is the most fascinating part of human physiology. If we could manipulate immune responses exactly like we wanted, infectious diseases, autoimmunity, and cancer would be problems of the past. However, we are very far from that goal.

BD: Describe the project for which you received the BD grant.

Thomas Haupt: My project focuses on biological treatments for Crohn's disease, a disorder of the gastrointestinal tract caused by an uncontrolled immune reaction against otherwise harmless gut bacteria. Biological treatments like infliximab, which targets tumor necrosis factor alpha, have revolutionized the treatment of Crohn's disease because they are highly effective. Unfortunately about one third of patients treated receive no benefit at all. We don't know why. Also, the drug's mechanism of action is not fully understood. We hypothesize that infliximab may specifically target and thereby increase the number of a certain type of T lymphocytes—called regulatory T cells—which are important for preventing autoimmunity.

By examining patients with Crohn's disease at baseline and weeks after they have initiated infliximab treatment, we hope to investigate the effect of infliximab on regulatory T cells. These cells may be counted by flow cytometry, and we will test their function by culturing them with normal T cells to see if the presence of one type of T cell inhibits the proliferation of the other. We think that the potential effect on regulatory T cells by infliximab may be caused by stimulation of a certain signaling pathway, specifically the TGF-beta pathway. It is possible to measure the activity in this pathway by staining the intracellular components of it with flow cytometry antibodies after fixing and permeabilizing the cells.

BD: What are the long- and short-term scientific goals of this project?

Thomas Haupt: In the short term, we hope to confirm or reject the hypothesis that infliximab treatment affects regulatory T cells by increasing the TGF-beta signaling. If so, it would be possible to predict the effect, or lack thereof, by simply measuring the number of regulatory T cells in patients. In the long term, we hope to reach a better understanding of the mechanisms behind lack of Infliximab response, which may hold the key to new treatments for these patients.

BD: What are the implications of your project for human health?

Thomas Haupt: Ultimately, we would like to get better at targeting expensive, potentially harmful treatments to patients for whom the greatest benefit is possible. We hope to obtain new knowledge about non-responders that will help us develop new, effective treatments for them.

BD: Which BD reagents do you plan to use, and for what purposes?

Thomas Haupt: We use BD CPT tubes for blood collection, since they allow for very easy separation of immune cells. To label and count regulatory T cells, we use the BD Treg cocktail of surface markers (anti-CD4, -CD25, and -CD127) as well as a FoxP3 antibody, which is the gold standard marker for detecting regulatory T cells. Also, we have included a CD161 antibody, which is a Th17 marker for pro-inflammatory T cells important in Crohn's disease, to get an idea about how treatment affects this compartment.

To investigate the TGF-beta signaling pathway, we will need to label the individual proteins in the cascade, the Smad2/3, p-Smad2/3 complexes, and Smad7. After stimulating the cells with recombinant TGF-beta1, we will surface-stain with the Treg cocktail, then fix and permeabilize them with the BD Phosflow™ buffer system to allow for intracellular staining. We will probably need to experiment with different combinations of primary and secondary antibodies, since there is no fluorochrome-conjugated Smad7 antibody currently available. However, the secondary antibodies will be from BD, and will be phycoerythrin conjugated for maximum brightness.


Veronika Kanderova, PhD
Postdoctoral Fellow
Charles University, Prague

Abstract Title:
High-content Affinity Proteomics Reveals the Response to Targeted Therapy Treatment in Childhood Acute Leukemias


BD: What is your educational background?

Veronika Kanderova: I received my master's degree in immunology from the Faculty of Science, and my PhD from the Second Faculty of Medicine, both at Charles University in Prague, Czech Republic. During my studies I worked in the research and development department of a Czech company that produced antibodies. I am currently in a postdoctoral fellowship at the CLIP (Childhood Leukemia Investigation Prague) laboratory at the Second Faculty of Medicine, Charles University and Motol University Hospital. My adviser is Tomas Kalina, MD, PhD, an expert in multicolor flow cytometry. Previously I held a UICC International Cancer Technology Transfer Fellowship in the laboratory of Dr. Fridtjof Lund-Johansen at Oslo University in Norway, where I gained expertise in his innovative flow cytometry-based proteomic method called SEC-MAP (size-exclusion chromatography - microsphere-based affinity proteomics).

BD: How and when did you become interested in science?

Veronika Kanderova: I grew up in the countryside and was raised to love nature. When I was a child, there was a French animated TV series for children, "Il était une fois... la vie," a show about the human body that I loved. I focused on science in high school, and by the time I began my university studies I had a clear vision: to be an immunologist.

BD: How did you become interested in your broad field of study?

Veronika Kanderova: Our immune system is why we survive on this planet full of pathogens and microbes. I have always been in awe, and at the same time enjoyed, the immune system's diversity and the interconnections between its many cells and molecules. Sometimes things go awry, however, as evidenced in leukemia. My favorite immune system cells are B cells. I am even a member of "Friend of B-cells Club." Immunology is a part of my life … of our lives.

BD: Describe the project for which you received the BD grant.

Veronika Kanderova: I work in CLIP, which is the leading clinical hematological laboratory in the Czech Republic. Our group focuses on childhood acute leukemia, the most common pediatric cancer. In the Czech Republic children are treated following progressive protocols of the International BFM Study Group. These treatment protocols are very effective but they are also associated with considerable toxicity. Moreover, approximately 20% of patients suffer a relapse. Recent successful development of targeted therapeutics in hematology opens the way for use of small molecular inhibitors and monoclonal antibodies for overcoming the chemoresistance and for potentially diminishing the toxicities associated with therapy. The critical problem is uncovering patients who can profit from these novel therapies. Molecular genetic techniques are powerful but they cannot address the precise mechanism of action of these modern targeted therapeutics. For this purpose we have introduced a novel proteomic array with algorithmic tools for high-dimensional data analysis of patient sample evaluation. SEC-MAP is an innovative flow cytometry-based platform that enables detection of more than 1,700 possible therapeutic targets and related molecules in a single sample. The method provides expression levels, relative amounts, subcellular localization, and characterizes molecular complexes. Moreover it allows for the detection of post-translational modifications, for example phosphorylation, that may indicate activation or deactivation of the signal transduction pathways connected with the response to therapy, or to resistance.

BD: What are the long- and short-term scientific goals of this project?

Veronika Kanderova: The short-term goal is developing the proteomic methodology for assessing targeted therapy in primary leukemia samples. We will create a SEC-MAP array that has the potential to reveal the precise function of targeted therapeutics, for example the tyrosine kinase inhibitor imatinib mesylate and the monoclonal antibody anti-CD22, at the protein level—that is, expression versus post-translational modification. Our long-term goal is to find appropriate correlates of good responses to these modern targeted therapies in current treatment protocols.

BD: What are the implications of your project for human health?

Veronika Kanderova: If successful, this approach should identify patients who can benefit from targeted therapy with no additive toxicity, within the standard treatment protocols. This can ultimately improve quality of life of children with acute leukemia.

BD: Which BD reagents do you plan to use, and for what purposes?

Veronika Kanderova: We plan to use BD antibodies against both phosphorylated and non-phosphorylated proteins involved in signaling pathways connected with targeted therapy response in childhood acute leukemia, for example transcription factors, kinases, apoptosis proteins, and others. Since SEC-MAP technology is based on immunoprecipitation, we will choose highly specific antibodies. The main phospho-proteomic changes will be verified with BD Phosflow™ antibodies using classical single-cell phospho-flow detection.


Manuel Martinez Garcia, PhD
Associate Professor
University of Alicante

Abstract Title:
Deciphering Unknown Human Viruses: A New Approach to Disentangle the Human Virome


BD: What is your educational background?

Manuel Martinez Garcia: I graduated with honors in biology from the University of Alicante in Spain. I then received an MSc in molecular and experimental biology, and my PhD in molecular microbial ecology, from the same university. In 2006, I joined the Division of Microbiology at Alicante as assistant lecturer of Microbiology. I have completed three postdoctoral fellowships in Europe: at the Max Planck Institute for Marine Microbiology (Bremen), Ludwig-Maximilians University (Munich), and the Laboratory of Microbiology (Academy of Sciences of the Czech Republic). During 2009 to 2011 I moved to United States, where I completed a postdoctoral fellowship at the Single Cell Genomics Center under the supervision of Dr. Ramunas Stepanauskas at Bigelow Laboratory, in Maine. There I developed techniques for single-cell genomics combined with metagenomics to study the metabolic capabilities of uncultured microbes in nature.

BD: How and when did you become interested in science?

Manuel Martinez Garcia: I found biology extremely fascinating in high school, especially DNA structure and replication mechanisms. I have very fond memories of my teacher, "the bearded man," who explained Okazaki fragments. From that point—I was 17 years old—I knew that science was for me and started to devour books on cosmology, Darwin's and Einstein's biographies, and others from my parents' library.

BD: How did you become interested in your broad field of study?

Manuel Martinez Garcia: Thanks to microbial ecology, we now know that most microbes in nature, including those living with us, cannot be replicated in a petri dish. Thus, we only know a tiny fraction of the extant diversity of microbes. Viruses, along with bacteria, are by far the most numerous and diverse biological entities in the human body. A typical human body contains 1,500 distinct virus types, and three trillion viruses all told. In recent years, microbes have taught us that they are not simple passengers in our bodies, but instead play key roles in our physiology, including our immune response and metabolism, as well as being responsible for some diseases. Recent clinical and experimental data has indeed widened our view on the role of human viruses as important factors that trigger or aggravate complex diseases, such as type 1 diabetes, asthma, irritable bowel disease, cystic fibrosis, and even certain cancers. Yet recent data suggests that we understand approximately one percent of viral genomics, which limits what we know about the biological roles of these viruses. The fact that most of these viruses are unknown inspired me to choose immunology, albeit from an unconventional perspective.

BD: Describe the project for which you received the BD grant.

Manuel Martinez Garcia: Traditionally, in medicine and immunology, standard cultivation techniques have been the gold standard for isolating and propagating human viruses. Virus culture relies on the ability to establish virus-host systems. But, as stated earlier, since roughly 99% of microbes, including viruses, remain uncultured and thus unidentified, we need to develop alternative strategies for bridging the gap between what we know and don't know regarding human virus biodiversity. Only then will we then understand the human "interactome," and how viruses affect bodily functions. Thus, in this project, we propose a new, radical alternative approach that combines pioneering flow cytometric and genomic techniques to obtain the genomic information from single, individual, uncultured human viruses. Towards that end, we will optimize advanced flow cytometry techniques to separate individual viruses from human samples by fluorescence activated virus sorting (FAVS) using a BD Influx™ cytometer-sorter. Once individual viruses have been separated and genetic material released from the viral capsid, we will amplify it to generate enough material for DNA sequencing.

BD: What are the long- and short-term scientific goals of this project?

Manuel Martinez Garcia: We plan on developing a new scientific approach that will allow us to map the viral genomes from samples as small as a single, individual virus. We expect to retrieve viruses directly from different parts of the bodies of healthy people and HIV patients. We believe our methodology will also be applicable to other scientific disciplines.

BD: What are the implications of your project for human health?

Manuel Martinez Garcia: Targeting two contrasting biological models—healthy people versus immunosuppressed patients—provides an opportunity to study the dynamics of the uncultured human virome. Ultimately, the significance is in identifying those predominant or rare uncultured, unknown viruses whose biological role remains enigmatic. By employing this novel approach, based on the DNA sequencing from single uncultured viruses, we will better understand the potential of certain viruses to trigger diseases such as type 1 diabetes and other illnesses.

BD: Which BD reagents do you plan to use, and for what purposes?

Manuel Martinez Garcia: Since this project involves isolating single viruses by flow cytometry, we plan to use BD reagents suitable for that instrument platform, including calibration beads, reagents for viral capsid and DNA staining, HIV antibodies, and reagents for HIV monitoring.


Marta Isabel Abreu Oliveira, PhD
Postdoctoral Fellow
Institute of Biomedical Engineering
University of Porto, Portugal

Abstract Title:
Dissecting Macrophage-Treg Crosstalk as a Therapeutic Approach for Colorectal Cancer


BD: What is your educational background?

Marta Oliveira: I received my BS in biology and my PhD in biomedical sciences, concentration in immunology, from the University of Porto. My PhD project involved molecular interactions at the T-cell surface. I then moved to the Institute of Biomedical Engineering (IBMC) at Porto as a postdoctoral fellow, to work on the inflammatory response to biomaterials and its impact on tissue regeneration. I've also undertaken cancer research that addresses the role of macrophages on cancer cell invasion, with the aim of designing new and more efficient anti-cancer therapeutic strategies.

BD: How and when did you become interested in science?

Marta Oliveira: I have always been a very curious person, eager to understand how and why different phenomena occur. In high school, biology classes were my favorite, so I guess I have been developing this interest in the origin and mechanisms of processes occurring in living organisms from an early age. For me, the idea of discovering something new is quite exciting, and being able to share and use that knowledge, towards a new therapy for example, is incredibly rewarding.

BD: How did you become interested in your broad field of study?

Marta Oliveira: After completing my biology degree, I joined the molecular immunology unit of IBMC to work on a tumor immunology-related project. During that time I had the opportunity of working with great scientists, particularly my mentors Prof. Alexandre do Carmo and Prof. Simon Davis, whose enthusiasm and excellence prompted me to continue with immunology. Also, the complexity of the immune system and its fine-tuned regulation have been a constant challenge. Understanding immune cell behavior under different contexts —physiological/ pathological—with the aim of modulating specific cellular responses in a therapeutic setting, has been the focus of my research ever since.

BD: Describe the project for which you received the BD grant.

Marta Oliveira: Tumor-associated macrophages (TAMs) and regulatory T cells (Tregs) infiltrate tumors and are considered promoters of cancer cell activities, contributing to disease progression. However, in colorectal cancer (CRC), the third most common cancer worldwide, their role is controversial because in these patients, increased intratumoral levels of TAMs or Tregs correlate with favorable prognosis, contrary to what occurs in most carcinomas. We propose to explore the Treg-TAM crosstalk in CRC as a potential target for future therapeutic strategies. We hypothesize that under the inflammatory tumor microenvironment, Tregs modulate TAM behavior, and both cell types work as allies against cancer development. But in CRC data on TAM density, location and, most importantly, phenotype, is scarce and contradictory. Moreover, the role of Tregs is unclear as is their impact on TAMs and the relevance of such modulation for CRC development. Thus, we propose to study Treg-TAM communication within this context and evaluate its impact in vivo for tumor progression and metastasis. Furthermore, we aim at dissecting the underlying molecular mechanisms.

To accomplish these goals, we will isolate monocytes from healthy blood donors and differentiate them into macrophages in the presence of Tregs and human CRC cells, on indirect co-culture systems. We will determine the phenotype of Treg-educated macrophages, and Treg/macrophage influence on cancer cell invasion and metastasis, using the chick-embryo model. We will also evaluate involvement of relevant signaling pathways. Importantly, prognostic significance of combined TAM/Treg infiltration in CRC will be investigated using fresh and frozen samples from CRC patients. Unraveling immunoregulatory mechanisms underlying disease progression will likely clarify disease pathogenesis.

BD: What are the long- and short-term scientific goals of this project?

Marta Oliveira: The hypothesis behind this proposal is that Tregs and macrophages, influenced by the CRC microenvironment, cooperate against tumor development. This argues against the existing paradigm of Tregs and macrophages in cancer, but it is supported by research, including our own results, and by clinical data. Through this work we expect to clarify the ability of Tregs to modulate macrophage function within the context of CRC, defining whether such regulation contributes to delay or even inhibits tumor development. These results will also highlight the role of the CRC ecosystem on immune cell regulation. Moreover, we expect to determine the relevance and involvement of EGFR-downstream signaling cascades on cancer cell invasion when both Tregs and macrophages are present.

BD: What are the implications of your project for human health?

Marta Oliveira: By identifying specific targets, we hope to contribute to more efficient therapeutic interventions. We also expect to correlate the prognostic value of combined TAM and Treg infiltration with their density and location within the different tumor regions, and to associate this data with patients' clinico-pathological data. This association may help to establish the role of Treg/macrophage crosstalk with CRC progression and schemes of treatment. Last, unraveling the function of these cells under the influence of the tumor microenvironment will offer great possibilities for shaping immune cell responses, opening great therapeutic perspectives.

BD: Which BD reagents do you plan to use, and for what purposes?

Marta Oliveira: We will use BD magnetic cell separation reagents to isolate Tregs and monocytes, and perform cell invasion assays with Matrigel® invasion chambers. In addition, BD antibodies will be used for flow cytometry, Western blotting, and immunohistochemistry, whereas cytokine secretion profiles will be characterized taking advantage of the BD™ Cytometric Bead Array (CBA) Flex Sets.

Cy™ is a trademark of GE Healthcare.

Matrigel is a registered trademark of Corning Incorporated.