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Academician Nicolae Cajal Symposium – 5Th Edition Translational Oncology, Cell Therapy, Bucharest, April 14–16, 2010

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Academician Nicolae Cajal Symposium – 5Th Edition Translational Oncology, Cell Therapy, Bucharest, April 14–16, 2010 ACADEMICIAN NICOLAE CAJAL SYMPOSIUM – 5TH EDITION TRANSLATIONAL ONCOLOGY, CELL THERAPY, BUCHAREST, APRIL 14–16, 2010 36 Translational Oncology Session Academician Nicolae Cajal Symposium 37 FOREWORD Professor Nicolae Cajal (1919–2004) is considered between the founders of a new Romanian scientific school, a research institute of the Romanian Academy called the “St. Nicolau” Institute of Virology. Many renowned physicians, pioneers of national medical specialties, have been educated in this Institute or in the Chair of Virology at the Bucharest University of Medicine and Pharmacy. Among Cajal’s most significant scientific contributions we will cite only a few, such as production and application of viral vaccines against poliomyelitis, influenza, and measles; important research programs in the domain of significant viral diseases as: AIDS, viral hepatitis, acute and chronic viral neuroinfections; the implication of certain viruses in oncogenesis and others. Cajal set high standards for him and other and shaped the field by training many young virologists all over the country and abroad. His experience indicates that successful projects adopt multidisciplinary approaches. Many of Cajal’s students and their second-, and third- “descendants” are still major contributors to the field of virology, vaccinology and cell biology. In the last years Cajal Foundation created by his daughter, Irina Cajal, organized annual scientific sympozia that have been attended by scientists in microbiology, biochemistry, informatics etc. as well as practitioners in the field of infectious diseases and epidemiolgy. This year symposium hosted three sessions: virology, genomics and translational oncology. 38 Translational Oncology Session TRANSLATIONAL ONCOLOGY SESSION DEFA-1, DEFA-5, DEFA-6, IL-8, CXCL-1, SPP-1, CTHRC-1), identified as up-regulated by microarray, in adenoma and tumoral samples FROM BENCH TO BEDSIDE: compared with matched normal tissues. NEW POSSIBLE BIOMARKERS IN EARLY AND LATE STAGES OF COLON CANCER - PRELIMINARY RESULTS CONCLUSION LILIANA LIVIA PASLARU1, ANCA NASTASE2, 3 3 All analysed genes are involved in tumorigenesis SIMONA DIMA , VLAD HERLEA , MIHNEA and progression. IONESCU3,CRISTIAN GHEORGHE3, TRAIAN DUMITRASCU3, SORIN ALEXANDRESCU3, ANA We remarked that DEFA5 and DEFA6 are key MARIA EFTIMIE2,ANA MARIA NICULESCU2, factors in adenoma formation while matrylisin 1 VLADIMIR LAZAR4 and IRINEL POPESCU3 (MMP7) is very important in the onset and in the 1 University of Medicine and Pharmacy „Carol Davila” progression of the disease. Bucharest, Postgraduate Department of Biochemistry The pattern of expression for the studied 2 RNTECH Bucharest, Romania cytokine shows that colon cancer begins and 3 Fundeni Clinical Institute of Digestive Diseases progress as an inflammatory condition. Thus, we and Liver Transplantation found out that IL8 has a high mRNA level in 4 Institute of Cancerology Gustave Roussy, Villejuif, France adenomas and irrespective of tumor stage, in adenocarcinoma as well. The level of expression INTRODUCTION for SPP1 is correlated with tumor level. Therefore within our study we underline the Today, an important priority in cancer research importance of using a panel of biomarkers for is the detection of new clinically useful biomarkers diseases evaluation and the implementation of the whose expression could represent important tools personalized treatment. for a better diagnostic, prognostic and for implementing personalized treatment of the disease. TUMOR-ASSOCIATED FIBROBLASTS INVOLVEMENT IN CANCER DEVELOPMENT MATERIAL AND METHODS PAUNESCU V.1, BOJIN FLORINA1, TATU C.A.1,2, The trasnscriptomic study was realized on a cohort of 31 GAVRILIUC OANA1, ROSCA ADRIANA2, GRUIA patients with primary colon tumors. ALEXANDRA2, CIUCULESCU FELICIA1, TANASIE We used triplets represented by normal, adenoma and GABRIELA1,2, TATU CARMEN1,2 and BUNU CARMEN1 tumoral tissue collected from the same patient during surgery. All these samples were analyzed by microarray and validated 1”Victor Babes” University of Medicine and Pharmacy by real time-PCR. Timisoara, Romania In accordance with national and international standards, 2 Clinical Emergency County Hospital Timisoara, Romania informed written consent was obtained from all patients and research protocols were approved by the ethical committee. INTRODUCTION RESULTS The complex cellular tumor (micro) environment comprises immunocompetent and inflammatory The microarray study allowed us to assess the cells, endothelial cells and fibroblasts. All of these candidate gene expression differences between cell types may critically influence the multi-step tumoral and normal tissue and between adenoma process of carcinogenesis and malignant phenotype. and normal tissue. Thus, we could identify Fibroblasts are known to take part in immune important genes involved in colon cancer reaction during tissue damage and injury by carcinogenesis and progression. modulating local cellular and cytokine milieu, In order to validate mRNA differential adjusting the kinetics and components of the expression, we analyzed by real-time PCR the inflammatory infiltrate, and by modulating the expression of 10 genes (MMP-1, MMP-3, MMP-7, functional status of the immunocompetent cells. Proc. Rom. Acad., Series B, 2010, 1, p. 37–56 Academician Nicolae Cajal Symposium 39 Main purpose of our study was investigation of deactivating/suppressing factors and neoangiogenesis tumor-associated fibroblasts (TAFs) phenotype and molecules should stimulate consideration of TAFs- function, and to what extent they become activated based therapeutic designs. within tumor environment and secrete different paracrine and autocrine factors, which will further Key words: Tumor-associated fibroblasts, play a role in tumorigenesis. chemokines, VEGF, tumorigenesis ACKNOWLEDGEMENTS MATERIAL AND METHODS The study was supported by PNCD-II Grant ONCOSTEM Human TAFs were isolated from breast cancer surgical no. 41-019/2007. pieces using enzymatic digestion, cultivated and expanded in vitro as monolayer cell culture. When cell population was pure, the trilineage differentiation potential (to adypocytes, MOLECULAR MARKERS IN BCR/ABL chondrocytes, and osteoblasts) was assessed under appropriate differentiation medium. Supernatants of cell cultures at NEGATIVE MYELOPROLIFERATIVE different passages were collected and presence of IL-4, IL-10, NEOPLASMS. FROM DETECTION TO IL-13, TGF-β1, TNF-α, INF-γ and VEGF was detected by SELECTIVE THERAPY ELISA method. Specific mouse anti-human fluorochrome- conjugated antibodies identified in flow-cytometric analysis CARMEN C. DIACONU1, LAURA DRAGOMIR NECULA1, surface markers of TAFs, such as CD14, CD117, CD90, MIHAELA CHIVU1, MIHAELA CLOSCA-GHEORGHIU2, CD106, CD44, CD29, CD73, HLA-DR, CXCR4. Expression ANCA R. LUPU2, AURORA ARGHIR3, GEORGETA of cytoskeleton and extracellular matrix proteins was revealed CARDOS3, ANA NEAGU1, IOANA ALDEA1, LILIA by immunocytochemistry (vimentin, α-smooth muscle actin). MATEI1, CORALIA BLEOTU1, IRINA ALEXIU1, Breast cancer isolated tumor cells and tumor cell lines (SK- DENISA DRAGU1, MARIUS ATAMAN1, COSMIN I. BR3, MDA-MB231, and MDA-MB468) were tested for their STANCU1, DELIA MUT-POPESCU2, AGRIPINA ability to adhere to the TAFs substrate in flowchamber when LUNGEANU3, MIHAIL E. HINESCU3 and STEFAN N. the shear stress generated was increasing from 0.35 to 15 CONSTANTINESCU4 dyne/cm2. Stimulation of TAFs used endogenous secretory factors for flowchamber assays. 1 Stefan S. Nicolau Institute of Virology, Bucharest, Romania 2 “Coltea” Clinic Hospital Bucharest, Romania 3 “Dr. Victor Babes” National Institute of Pathology, Bucharest, Romania RESULTS 4 Université Catholique de Louvain, Brussels, Christian de Duve Institute of Cellular Pathology, Ludwig Institute for Secretion of cytokines with direct immuno- Cancer Research, Belgium suppressive effect, such as IL-10 and IL-13, and other cytokines, IL-4 and TNF-α, was increased in cultured TAFs at all passages. Enhanced INTRODUCTION production of TGF-β1 by TAFs may also be a critical factor in tumor homeostasis. Expression of The main BCR/ABL negative myeloproliferative MHC class II molecules (HLA-DR) on activated neoplasms (MPN) are today recognized as TAFs contribute to an additional level of Polycythemia Vera (PV), Essential Thrombo- immunosuppression and pro-tumoral effect. High cythemia (ET) and Primary Myelofibrosis (PMF). level of VEGF production suggests that TAFs These conditions are clonal disorders of provide not only structural support, but also pro- hematopoietic progenitor cells that are able to angiogenetic molecules for tumor vascularization. differentiate into excess numbers of mature cells. α-smooth muscle actin expression in fibroblasts is The recently discovered JAK2 V617F mutation has variable and associated with an activated status. become the biological marker for these conditions Adherence of tumor cells on TAFs substrate was and promoted the development of selective therapy. increased when stimulated with VEGF for all JAK2 V617F or other JAK2 JH2 region mutations values of shear stress, while IL-4 induced lead to a lack of inhibition of the JH1 domain and increased adherence only for shear stresses less enables constitutive tyrosine kinase activity. than 8 dyne/cm2. Recently, mutations in the thrombopoietin receptor (c-Mpl) that act by causing activating conforma- tional changes in the receptor, have been found. CONCLUSION The more precise diagnosis of the patients will lead to advances of clinical investigations
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