19^th Euro Congress on Cancer Science and Therapy July 17-19, 2017 Lisbon, Portugal

Nyla A Heerema is a professor in the Department of Pathology at The Ohio State University Wexner Medical Center. She is the Director of the Cancer Cytogenetics Laboratory there, as well as conducts an active research program. Her areas of research are the cytogenetics of chronic lymphocytic leukemia (CLL) and of pediatric acute lymphoblastic leukemia. She is a member of the CLL Research Consortium, and is the Chairperson of the Cytogenetics Discipline for the Children’s Oncology Group. She has published over 300 articles.

CLL has a variable clinical course and prognostic factors are vital. Metaphase cytogenetics have been minimally informative as CLL cells do not respond to traditional mitogens. CpG stimulates CLL cells to divide in <80% cases. Investigation of karyotypic abnormalities within one year of diagnosis in untreated CLL patients using CpG-stimulation identified complex karyotype (CK) (>3 unrelated abnormalities) that predicted a shorter time to first treatment (TFT) compared to non-CK (NCK, 12 months estimates 45% and 15%, respectively, p=0.0005). Despite a strong correlation of del(17p) with CK, CK predicted TFT independent of del(17p), a known poor prognosticator. Additionally, in patients with either balanced or unbalanced translocation, the good prognosis of mutated IGHV was negated (mutated IGHV translocation present vs absent, HR 3.59, p<0.001; unmutated IGHV translocation present vs absent, HR 1.03, p=0.92, interaction p=0.002). Independent of IGHV and translocation, CK (HR 1.70, p=0.037) remained statistically significant. Patients with Richter’s transformation (RT), an aggressive lymphoma in some CLL patients, exhibited higher risk for death with CK (HR 2.72, p=0.025) than in patients with NCK after R-EPOCH treatment. CK was independently associated with ibrutinib discontinuation due to progression. Although a low percentage of patients treated with ibrutinib experience RT, 6/9 patients with near-tetraploidy detected prior to ibrutinib treatment developed RT. In a multivariable model, both near-tetraploidy (HR 8.66, p<0.0001) and CK (HR 4.78, p=0.01) were independent risk factors for discontinuing ibrutinib due to transformation. In conclusion, CpG-stimulated karyotypes should be performed in CLL patients to identify karyotypic abnormalities that are significant for prognostication.

Charles L Hitchcock is an Emeritus Faculty member of the Department of Pathology at The Ohio State University in Columbus, Ohio USA. For the last 35 years, he has been a member of a multidisciplinary team of physicians, engineers, chemists, and biomedical scientists whose goal is to provide physicians with the resources to optimize the treatment of patients with solid tumors. Their efforts have resulted in over 500 publications and abstracts, multiple research grants, patents, and several biotech companies.

Current imaging and exploration of the surgical field using inspection and palpation provides the surgeon with limited informationfor clinical decision making. Using colorectal adenocarcinoma as a model, we developed a “system” that incorporates currently available technologies to increase the precision of tumor imaging before and during surgery. The system has three parts: 1) a tumorrelated marker, 2) a labeled molecular probe, and 3) instrumentation for imaging and/or detection of the labeled tumor. The glycoprotein TAG-72 is expressed in more than 80% of adenocarcinomas of the: colon and rectum, pancreas, lung, prostate, endometrium, and ovary. We utilized three different generations of 125I-labeled murine IgG antibodies to TAG-72. Our earlier studies used a hand-held gamma probe for the detection and excision of TAG-72 positive tissues. Follow-up studies clearly demonstrated that, independent of the pathologic stage, patient survival was significantly better if all of the TAG-positive tissue was removed. In the last 25 tears the “system” has evolved to the use of low energy labeled, small, molecular probes that allow for combined intraoperative imaging and detection. We bioengineered humanized, single chain, fragments and its “mers”, to TAG-72 that minimize murine antibodies associated problems. 123I-labeling of these small molecules provides an optimal signal-to-noise ratio. The “System” now allows for preoperative SPECT/CT imaging to determine actual disease extent prior to surgery and for using a portable gamma camera for realtime intraoperative imaging while retaining hand-held-gamma probe detection of TAG-72 positive tissue. Proof-of-concept studies

clearly demonstrate that the surgeon can “get it all”.

Alice Dragomir is an Assistant Professor at the McGill University, Faculty of Medicine, and Scientist in Health Economics and Outcomes Research at the Research Institute of the McGill University Health Center. She is an Economist and Biostatistician with Master’s degree in Statistics and Doctoral degree in Pharmacoepidemiology and Pharmacoeconomics, from University of Montreal, Canada. She has 14 years of experience in academic research. She was involved in research projects focused on evaluation of health outcomes and health economics related to different treatment strategies, adherence to treatments, health services utilization and disease modeling. Her current research is focused on clinical and economic evaluation of different treatments strategies offered to patients with prostate cancer or other urologic cancers. She has an extended experience in analyzing administrative healthcare databases and disease modeling. Her research represents a valuable tool for decision-makers and clinician leaders while evaluating the clinical and economic impacts of innovative treatments.

Significant advances have been made in the field of urological cancers, including new health technologies for localized cancers and many innovative drugs for advanced cancers. These novel treatment options are generally more expensive than existing treatments which limits their implementation in current clinical practice. Particularly, several new tests have demonstrated clinical utility and benefits in the screening, diagnosis or treatment phase of prostate cancer. Unfortunately, none of these tests are currently used routinely in clinical practice in Canada or other countries. One of the reasons is the lack of evidence regarding their cost-effectiveness. Recently, the Canadian Task Force on Preventive Healthcare and other similar entities from other countries, recommended against PSA screening in all men. We believe that applying this recommendation regardless of risk- and age-stratification to all men is an extreme strategy, especially considering the lack of economic evidence. An alternative needs to be found in a more proficient way to perform prostate cancer screening, diagnosis and treatment. Identifying the new prognosis/risk assessment tests or interventions which are cost-effective could be a first step in achieving this goal. Better prostate cancer risk assessment tools will assist the decisionmaking process, which would finally lead to the best therapeutic options being offered to individual patients, improve overall patient

care, and reduce healthcare expenditure. This presentation presents results of several studies aimed to determine the best solutions for urological cancer screening, diagnosis and treatment for improving healthcare delivery to patients, increase access to these new advances, while uncovering better ways to optimize health services and cost allocation.

M Helena Vasconcelos is an Assistant Professor at FFUP (Faculty of Pharmacy, University of Porto) and Group Leader of the Cancer Drug Resistance Group at i3S/ IPATIMUP. She has done her First degree in Pharmaceutical Sciences (1991) from FFUP in Portugal, MSc (1992) and PhD (1996) from the University of Aberdeen in Scotland. Her current research focuses on the identification and validation of biomarkers and therapeutic targets to overcome drug resistance in cancer and on the activity of small molecules to counteract drug resistance. She has published more than 85 papers published in international journals and with an h-index of 23.

Multidrug resistance (MDR) is often responsible for treatment failure in cancer patients. One of the main reasons for the MDR phenotype of cancer cells is an overexpression of drug-efflux pumps such as P-glycoprotein (P-gp). Recently, my research group, together with international collaborators, compared MDR cells with their drug-sensitive counterparts and verified that MDR cells present metabolic alterations which may be further explored as molecular targets to counteract the MDR phenotype. In addition, my group observed that P-gp may be horizontally transferred by extracellular vesicles (EVs), between MDR and drug-sensitive cells, confirming results from other researchers and indicating that this protein has a stronger influence in the MDR phenotype of tumor cells than had been initially realized. Interestingly, we and collaborators had recently found that the EVs released by MDR cells are enriched in microvesicle-like EVs. However, the work indicates that drug-resistant cells without overexpression of P-gp do not present this enrichment. Thus, we are currently verifying if P-gp could be involved in the release of microvesicles by MDR cells.

Audrey Claing has completed her PhD from the University of Sherbrooke (Canada) in 1997. She has worked at the Laboratory of Dr. Robert J Lefkowitz, 2012 Nobel Laureate, for her Post-doctoral studies at Duke University (USA). She is now the Professor of Pharmacology and Physiology at the Montreal University (Canada). She has published more than 60 papers in reputed journals and has greatly advanced research in the field of G Protein Signaling.

Triple-negative breast cancers (TNBC) are a highly invasive type of breast cancer and associated with poor prognostics. Although, these tumors do not express the typical hormone receptors (ER-, PR-), nor the HER2 receptor, their proliferation and invasive capacities can be enhanced by growth factors such as the epidermal growth factor (EGF). Drugs inhibiting EGF receptor activity have however shown limited effects mainly due to the development of resistance. There is therefore an urgent need to identify new therapeutic targets for the design of therapies that would complement current approaches (surgery, chemo and radiotherapy). We and others have shown that the Ras-related ADP-ribosylation factors (ARF) are another class of small GTPases regulating key features of cancer cells. ARF1 and ARF6, two isoforms best characterized, are highly expressed in cells and tumor tissue of the most aggressive and advanced subtypes of breast cancers. Knock down of ARF1 expression, for example, impairs the ability of breast cancer cells to proliferate, migrate and degrade the extracellular matrix. Furthermore, growth of primary tumors as well as lung metastasis is reduced in a murine xenograft model when expression of the GTPase is inhibited. Our findings have demonstrated that increased levels of ARF1, in non-invasive cells lead to the epithelial-mesenchymal transition (EMT). Overall, our work has identified ARF1 as a molecular switch of cancer progression and thus suggests that limiting the expression/activation of this GTPase could help improve outcome for breast cancer patients.

Hirendra Nath Banerjee completed his undergraduate degree in Biology & Chemistry and Bachelor of Medicine and Surgery degree from Calcutta University, India. His M.S. in Molecular Biology is from LIU at NY, USA and Ph.D. from Howard University Cancer Center in Washington, D.C. Dr. Banerjee did his post-doctoral training at Yale University and Medical University of South Carolina, USA. As a tenured Professor at Elizabeth City State University under the University of North Carolina system, Dr. Banerjee is involved in cancer research for more than two decades training many talented under graduate and graduate students in the process.

The recurrence and the metastasis of prostate cancer PCa are tightly linked with the biology of prostate cancer stem cells or cancer-initiating cells that is reminiscent of the acquisition of epithelial to mesenchymal transition (EMT) phenotype. Increasing evidence suggests that EMT-type cells share many biological characteristics with cancer stem-like cells (CSCs). In earlier studies it has been shown that there are certain genes and miRNA that are involved in this process. The genes and the miRNA have altered functionality causing this problem. We collected several prostate cancer samples from African American (AA) and Caucasian (CA) patients from Karmanos Cancer Center, Detroit, MI and compared the differential expression of those previously mentioned CSC

related miRNA in these samples searching for biomarkers with clinicopathological variables, including race (AA vs. CA), to develop a clinically relevant algorithm for PCa aggressiveness. We identified that the over expression of genes Lin28B and EZH2 leads to the acquisition of the invasive characteristics of the Pca cells with EMT phenotype due to down regulation of miR200 and let7c, there by maintaining the stem cell like characteristics in PCa which in turn causes more aggressive PCa in AA than CA along with up regulation of the PRC2 (polycomb) protein complex expression. Work proposed in this application, will contribute to development of a novel theranostics approach involving the noncoding RNA’s that may translate into an effective treatment regimen against not only PCa but other cancers as well.

Katarina Jeremić attended Medical School, University of Belgrade in 1996, MD in 2000, PhD in 2006 and Academic Special Studies in Gynecology and Obstetrics.She has 19 years of Clinical Experience, working as Gynecologist at Clinic for Gynecology & Obstetrics Clinical Centre of Serbia, which is the biggest one in whole region. She is currently the Head of Gynecologic Oncology Department and also member of many scientific projects on Cancer and Pregnancy. She worked at the Medical Faculty, University Belgrade as Lecturer and Associate Professor of Gynecology and Obstetrics. She has 50 publications in CC/SCI expanded and JCRindexed, and participated in more than 50 international congresses, with a total number of 150 publications. She is a member of FIGO, ESGO, and other societies.

Conservative approaches of early-stage endometrial carcinoma includes hormonal therapy, in selected group of young patients with endometrial carcinoma with age less than 45 years and wishes to preserve fertility, that shows low grade 1 endometrioid adenocarcinomas limited to the endometrium with MRI excluded myomaterial invasion, without evidence of limphovascular space involvement or extrauterine disease (cervical, ovarian, lymphnodal or any other extra-uterine disease). The diagnosis is proven by two experienced gynoncology pathologists review of analyzed endometrial samples. It could be collected by biopsy, hysteroscopy or dilatation & curatage and if it possible PgR analysis should be done. Invasive procedures for collecting endometrial samples are hysteroscopy that permits evaluation of endometrial cavity and biopsy of suspicious lesion. The accuracy of hysteroscopy is high with sensitivity rate of 86.4% and specific rate of 99.2%, even higher in the diagnosis, than in excluding it. The possible dissemination of malignant cells through fallopian tubes during hysteroscopy, has not been proven in meta analysis in early stage of the disease. Obligate pretreatment assessments include biopsy, hysteroscopy or dilatation & curatage, radiologic imaging, contrast MRI (to exclude myometrial invasion, exclude extrauterine spread of disease, ovarian, lymphonodal, cervical involvement) even laparoscopy and assessment of ovaries, peritoneum and PW + SLN, as also CA 125, X ray for chests examination. The results according to many studies are that almost a two third of patients (50-75% of patients) that are treated with gestagen therapy have complete response, but 20-45% patients will have recurrence even after initial response and 25% would not answer on the therapy. Follow up is repeating of endometrial biopsies by hysteroscopy every 3 months which is recommended, until there is a complete response or achieving pregnancy. Surgery is recommended if there is no response after 6 months of medication treatment. Hormonal therapy that could be applied is progestins that inhibits the estrogenic effect and suppresses cell proliferation (medroxy progesterone acetate, megestrl acetate), GnRh analogues, but also local gestagens ( IUD), oral natural progesterons, aromatase inhibitors - antiestrogens as also three step endoscopic (hysteroscopic) resection - remove tumour, surrounding endometrium, myometrium.

Vasco Fonseca has a degree in Medicine from the University of Lisbon in 2000. He is trained as a Medical Oncologist at the IPO of Lisbon and is currently working at the CHLO, in “Maria José Nogueira Pinto” Centre, as well as for the Portuguese National Military Forces. He is currently developing clinical trials in the area of breast cancer. He is the author of clinical protocol for Breast Cancer of the CHLO and has publications in reputed journals.

In the CHLO Breast Unit, which incorporates 4 hospitals, we treat almost 300 new breast cancer patients per year. In most cases, the international guidelines are very clear regarding locally advanced breast cancer, allowing a vast number of options in the grey area that concerns the Unit’s experience. We consider that the Unit’s experience and tumor staging, but also tumor biology, patient’s preference, individual risk factors and relative contraindications should be the principal considerations for the neoadjuvant treatment decision (according to international data). For this reason, we have formulated an internal protocol which allows us, not only to include all the indicated patients, but also to compile a database for their follow-up. In our protocol, triple negative, Her2 positive tumors and luminal B-like with high proliferative index, equal or above 2 cm (cT2N0), are proposed for neoadjuvant treatment. Patients with hormone dependent tumors that refuse surgical treatment, older patients, or patients with severe comorbidities, as well as selected luminal-A-like patients, are included in neoadjuvant hormonal treatment, which in some cases is extended over 8 months.

Shanmugasundaram Ganapathy Kanniappan obtained his PhD degree from the University of Madras, India and underwent Post-doctoral training at the National Institute of Immunology (NII, New Delhi), University of California at Los Angeles (UCLA) and Johns Hopkins University (USA). Currently, he is an Assistant Professor at the Johns Hopkins University School of Medicine. He has been a recipient of Research Fellowship/Lectureship, (CSIR-UGC, India), a Concept Award (Department of Defense (DoD), USA), and a Pilot Research Grant (SIR, USA). He has several publications, and serving as an active Editorial Board Member and reviewer for prestigious journals.

Our understanding of aerobic glycolysis or tumor glycolysis popularly known as the Warburg effect has significantly advanced in the past two decades. Besides facilitating tumor growth and survival under unfavorable conditions, aerobic glycolysis also plays a pivotal role in therapeutic resistance. For example, the extracellular accumulation of lactate, a product of glycolysis impacts the tumor microenvironment (TME) and contributes to its low-pH and acidity. Noteworthy, TME acts as a barrier that blocks the efficacy of anticancer agents including chemotherapeutics and immune therapeutics. Furthermore, aerobic glycolysis has also been ascribed to support the immune-evasion of cancer. Thus, tumor glycolysis which is an integral component of the metabolic reprogramming (hallmark of cancer) is linked with immune-evasion (another hallmark of cancer). Using preclinical models, we demonstrated that deregulation of tumor glycolysis and/or the induction of metabolic stress sensitizes cancer cells to natural killer (NK) cell-cytotoxicity. Mechanistically, metabolic perturbation up-regulated the stress-inducible ligands that are recognized by specific NK cell receptors for further elimination. Notably, our recent data also show that sub-lethal, metronomic treatment with current, clinically relevant chemotherapeutics may also up-regulate such stress-inducible ligands indicating their sensitivity to NK cell mediated cytotoxicity. Taken together, these findings reiterate that cancer metabolism remains a viable therapeutic target which could be exploited for both chemo- and immunotherapies.

Qi-Fei Wang has completed his Master’s degree from Dalian Medical University and is a PhD candidate at Tianjin Medical University. He is the Supervisor of Postgraduate students at the Dalian Medical University. He is a member of Rehabilitation Medical Association of Liaoning province China. He has published more than 12 papers in reputed journals.

Objective: The aim of this study was to evaluate application of “zero-ischemia” and “sutureless” techniques in laparoscopic partial nephrectomy (LPN) with holmium laser, and to question this technique in terms of reduction of warm ischemia time (WIT) and reduced renal tissue damage.

Methods: According to our experience of LPN, we want to explore a novel choice of operation that could avoid ischemia of kidney and reduced renal tissue damage. “Zero-ischemia” means don’t clamp the renal artery in operation; “sutureless” means that don’t stitch up the wound of kidney to hemostasis with surgical suture, but singe it using holmium laser.

Results: On the basis of our preliminary work about “zero-ischemia” and “sutureless” techniques in LPN with holmium laser, and combined with domestic and foreign literature reports. “Zero-ischemia” could reduce WIT to the greatest extent, avoid the damage of renal function, “sutureless” would reduce renal tissue damage region, The purpose of holmium laser, on the one hand it can stop bleeding instead of stitching up would with surgical suture, on the other hand it can reduce renal tissue damage region compare to traditional suture method.

Conclusions: Combined zero-ischemia and sutureless with holmium laser, this promising technique could avoid the damage of renal function as much as possible, and reduce renal tissue damage region to the greatest extent. The technique is feasible and secure under the premise of compatible indication, it can reduce renal tissue damage region and shorten operation time, and WIT will be reduced or even eliminated.

Adnan Yousif Rojeab has done his Doctorate in Electrical Engineering; he is a University Lecturer and Researcher. He has also worked at the Queen Mary University of London, University of West London. He is currently working at the London College. His research interests include action of electromagnetic fields on human system.

Cancer is a phenomenon of special reaction mechanism, which is functioning against the threat of severely acted tissues in the body. It is only created to eliminate severe damages and dangers that occur in the cells of the body, when the immune system fails to cure the damages. The effect of the cancer could only be ended when a direct, right and simple treatment method should be applied to cure the previous diseases that have caused the cancer, but not to attempt to treat the cancer itself. For the telomere matter, cancer cells have similar lengthening characteristics to those of germ cells. While in somatic cells, the telomere is shortening in every DNA replication. By applying a suitable amount and direction of a magnetic action on opposing the cancer cells, the lengthening of the telomere of the cancer could be inhibited, towards the somatic cells characteristics, and this application could be accepted as a method to treat cancer.

Adnan Yousif Rojeab has done his Doctorate in Electrical Engineering; he is a University Lecturer and Researcher. He has also worked at the Queen Mary University of London, University of West London. He is currently working at the London College. His research interests include action of electromagnetic fields on human system.

Cancer is a phenomenon of special reaction mechanism, which is functioning against the threat of severely acted tissues in the body. It is only created to eliminate severe damages and dangers that occur in the cells of the body, when the immune system fails to cure the damages. The effect of the cancer could only be ended when a direct, right and simple treatment method should be applied to cure the previous diseases that have caused the cancer, but not to attempt to treat the cancer itself. For the telomere matter, cancer cells have similar lengthening characteristics to those of germ cells. While in somatic cells, the telomere is shortening in every DNA replication. By applying a suitable amount and direction of a magnetic action on opposing the cancer cells, the lengthening of the telomere of the cancer could be inhibited, towards the somatic cells characteristics, and this application could be accepted as a method to treat cancer.

Robert Thomas is a Professor of Biological and Exercise Science at Coventry University and a Clinical Teacher at Cambridge University. He leads a research

team of oncology nurses and doctors at the Primrose Unit Bedford Hospital, which has an academic interest in the evaluation of nutritional, lifestyle and self-help strategies after cancer. The team has over 50 studies including the world’s largest randomized evaluation of a polyphenol rich nutritional supplement. He is an Editor of the lifestyle and cancer website (cancernet.co.uk) and the general heath website (keep-healthy.com) and designed the 1st UK approved course and qualification in cancer exercise rehabilitation. For his efforts to improve the long term wellbeing of patients, he was awarded the British Oncology Association Oncologist of the Year and Hospital Doctor Magazine UK Doctor of the Year.

As the chance of surviving, or achieving long term control, after cancer is improving, the number of people living with the consequence of cancer treatments at any one time is dramatically increasing. In the UK alone, this number has increased from two-three million in just five years. As consequences, the case for developing effective, self-help initiatives has never been stronger. This talk summarizes the international evidence, which proves that physical activity, nutrition and other lifestyle strategies can substantially reduce the risk of developing cancer, improve outcomes and ameliorate many of the long toxicities of consequences of biological therapies, chemotherapy and hormones and radiotherapy. These include fatigue, hot flushes, weight gain, arthritis, osteoporosis and mood disorders. This talk also highlights the underlying biological processes that take place in the body after a healthy lifestyle, which can have direct and indirect anti-cancer effects. By looking only at the scientific evidence, it breaks down the myths behind which foods to avoid and which to eat more of including nutritional interventions. It highlights the results of the world’s largest double blind randomized study of the National Cancer Research Network (NCRN) evaluation of a polyphenol rich nutritional intervention (The Pomi-T study). It emphasizes the potential risks and benefits of mineral and vitamin supplements to boost a healthy diet.

Sonia A Melo has completed her PhD from the Faculty of Medicine of University of Porto and Post-doctoral studies from Harvard Medical School and MD Anderson Cancer Center. She is a Principal Investigator at I3S - Institute of Research and Innovation of the University of Porto and Institute of Molecular Pathology and Immunology of Porto University. Her career is fully committed to cancer research and has published more than 20 papers in reputed journals.

Cancer treatment experienced significant advance over the last years. This has been mainly due to therapy targeting of key biological mechanisms such as cell survival, proliferation, apoptosis, angiogenesis and, more recently, tumor immune response. Cell communication, however, despite playing a fundamental role in all steps of tumor progression, up to metastasis and therapy resistance, is still off the cancer therapy landscape. Hence, the major challenge of our work is to bring cell communication into the realm of cancer therapy. Exosomes, extracellular vesicles secreted by all cells, are key mediators of cell to-cell communication in cancer. We have identified a network of communication mediated by cancer exosomes in vivo, through the identification of their recipient cells. Additionally we show that targeting the most frequent pancreatic cancer exosomes communication route can be an effective target for pancreatic cancer therapy. Our strategy includes unique genetically engineered mouse models (GEMMs), which allow for the first time to trace exosomes in vivo and model therapeutic targeting of exosomes biogenesis. Our work demonstrates the in vivo significance of pancreatic cancer exosomes and opens a new avenue for groundbreaking discoveries in the treatment of pancreatic cancer and, eventually, other cancers.

Zofia F Bielecka is a Medical Biotechnologist, currently working on tyrosine kinase inhibitors resistance in renal cell carcinoma. She is a PhD student at Warsaw Medical University, Poland (International School of Molecular Medicine). She is also a Research Assistant at Military Institute of Medicine in Warsaw, Poland and works in Laboratory of Molecular Oncology. She has gained considerable experience at the University of York, England and at the Lund University, Sweden. She has published 7 papers in reputed journals and has been serving a Reviewer in Elsevier journals. She is also a co-author of one patent application.

Primary-like resistance phenomenon to sorafenib (TKI) was previously observed in hypoxic conditions in two 3D models as well as in one 2D model in HKCSCs (human kidney cancer stem-like cells) cell line (Celprogen, cat no. 36117-44). Herein, we investigated the molecular background behind this phenomenon in 12 hypoxic total protein samples (each in triplicate) of renal cancer cells: primary HKCSCs resistant to sorafenib in hypoxia and other cell lines sensitive to sorafenib treatment in the same conditions: parental HKCSCs cell line, primary and metastatic renal cell carcinoma (RCC) cells from Memorial Sloan Kettering Cancer Centre (SK-RC-44 and SK-RC-45), metastatic RCC cell line (pleural effusion) ACHN cell line and HEK-293 cell line as a control. Additional control condition was untreated adequate cell lines in hypoxia. The acquired MS/MS data were pre-processed with Mascot Distiller software (v. 2.5, Matrix Science), and a database search was performed using the Mascot Search Engine (Matrix Science, Mascot Server 2.4.1) against the Swiss-Prot database restricted to human proteins. The results revealed potential candidates as elements of molecular mechanisms of primary resistance to sorafenib in renal cell carcinoma; those results will be further confirmed using Western blot analysis, possible confirming that apart from impact of hypoxia, the addition of targeted therapies alter signaling pathways in renal cancer cells, which means that mechanisms other than those involving hypoxia-inducible factors (HIFs) may be responsible for TKI-resistance.

Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) is a tumor suppressor and a negative regulator of different receptor tyrosine kinases (RTKs). The molecular mechanisms whereby LRIG1 mediates tumor suppression and regulates RTKs remain incompletely understood. Here, we performed a yeast two-hybrid screen to identify novel LRIG1-interacting proteins and datamined the BioPlex protein interaction data repository. All the putative LRIG1-interactors identified were functionally evaluated in a triple co-transfection system where HEK293 cells were co-transfected with platelet derived growth factor receptor alpha (PDGFRA) and LRIG1 together with shRNAs against the identified LRIG1-interactors.We took advantage of the ability of LRIG1 to downregulate the PDGFRA expression levels and evaluated the effects of the shRNAs against different LRIG1-interactors on the resulting PDGFRA levels. Based on the protein interaction data and our functional results, we propose a functional LRIG1 protein interaction network that contained mostly novel and unanticipated components. The network contained the functionally important LRIG1- interacting proteins RAB4A, PON2, GLRX3, GAL3ST1, TUBB8, ZBTB16, LRIG2, CNPY3, HLA-DRA, GML, CNPY4, LRRC40, and LRIG3, together with PTPRK and other proteins. In silico analyses of the cancer genome atlas (TCGA) data sets revealed consistent correlations between the expression of the transcripts for LRIG1 and its inter-actors ZBTB16 and PTPRK, and inverse correlations between the transcripts for LRIG1 and GLRX3. PON2 was further experimentally studied and found to co-localize with LRIG1 in

LRIG1 transfected cells. We suggest that the proposed LRIG1 protein interaction network will provide important novel leads for future studies aiming at understanding the molecular functions of LRIG1 and its tumor suppressive functions.

Carolina F Ruivo has completed her Master’s in Biomedical Engineering from IST, Lisbon in collaboration with University of Twente, Netherlands. Currently, she is working on a pre-doctoral project about the role of exosomes in tumor heterogeneity at Professor Sonia Melo’s Lab, Genetic Dynamics of Cancer Cells Group at University of Porto, Portugal.

Tumors are known to be heterogeneous, containing different cell types with distinct genetic and phenotypic features. Pancreatic cancer is a well-known example of tumor heterogeneity, showing a hierarchical organization of subpopulations with different tumorigenic potential. Intercommunication between tumor subpopulations is the key to development, progression and metastasis processes. Recent evidence shows that exosomes are important mediators of intercellular communication both at local and distant sites. If and how exosomes play a role in subpopulation communication is still unknown. In our work we have identified five subpopulations of pancreatic cancer cells based on cell surface markers which discriminate cells with different tumorigenic and self-renewal capacity. Using stable clones of cancer cells that express exosomes markers fused with fluorescent reporter proteins and secrete color-coded exosomes, we have studied the flow of exosomes between distinct subpopulations of cancer cells. Here we show that subpopulations of cancer cells communicate with each other via exosomes through an organized dynamic communication network (ExoNet). The presence of multicolor positive cells showed that exosomes are exchanged between different cancer cell subpopulations forming distinct routes of communication. The ExoNet reshapes in the presence of therapy to allow the tumor to respond and overcome the challenge. We have demonstrated that subpopulations of cancer cells communicate between them in a non-random manner using exosomes, and form a dynamic network of communication, which conveys the tumor with plastic properties that allows it to adapt in response to therapy.

Nuno Bastos has completed his BSc in Biochemistry in 2014 and MSc studies in Medicine and Molecular Oncology, last year, both in Porto University. During the

BSc and MSc studies he was in Porto´s IPO researching the role of microRNAs in therapy response and resistance in renal cell carcinoma and its aplicability as biomarkers. Recently, he joined the Genetic Dynamics of Cancer Cells at I3S aiming to perform a PhD under the supervision of Professor Sónia Melo.

Pancreatic cancer (PDAC) represents one of the most lethal cancers mainly due to a lack of reliable therapeutic options. Cell communication, in spite of playing a major role in tumor progression, is still off the cancer therapy landscape. Exosomes, extracellular vesicles derived from the endocytic pathway, are an important cell-to-cell communication system with neighbor/ distant cells. Our main aim is to study the role of exosomes biogenesis during PDAC progression and understand if cancer exosomes biogenesis could be a new therapeutic target in PDAC. Rab GTPases are crucial proteins in exosomes biogenesis and are involved in all stages of the endocytic pathway. We show that during PDAC progression Rab-5, -7, -27a and -27b are differently expressed. Increased expression of Rab-27a and -27b correlates with an increase in exosomes number, and these features are associated with a more aggressive phenotype. Additionally, when treated with, the standard care chemotherapeutic for PDAC, cancer cells change their exosomes biogenesis pattern, increasing exosomes release. Finally, we are using an inducible and conditional genetically engineered Rab27a knockout mouse model that spontaneously develops PDAC, to study the role of exosomes and its biogenesis in disease progression and therapy response, evaluating exosomes mediated communication as a new therapeutic option in PDAC.