Objectives (research directions)
The Biomedical Applications Unit launched its activity in 1996 and for over a decade has been specialized in exploiting the basic research results disclosed by the IBMCB for the development of biomedical products and services applicable to the field of invitro diagnostics. It has recently extended its activities towards the research of stem cells. More specifically currently, a) the BAU has characterized and compared the properties of human stem cell populations derived from different tissues and b) has established the optimal candidates for cancer therapy, differentiation and studies on the toxicity of drugs.
Human stem cell research
Stem cells are undifferentiated cells with a capacity for clonal expansion and differentiation into multiple functional cell types in vitro and in vivo. The research interests of BAU in the field of stem cells focus on the investigation of the mechanisms governing human post-natal stem cell differentiation in response to stimuli from their microenvironment and their potential use for regenerative medicine purposes. A second area of interest is the characterization and investigation of the differences in the biology of mesenchymal stem cell populations derived from different anatomical sites and developmental stages. Furthermore, the research team of BAU is investigating the use of these populations for cancer cytotherapy in vitro and in animal models and in protocols for differentiation and drug toxicity.
Other research interests of the Unit aim at identifying new prognostic markers and therapeutic targets for cancer. Additionally the objectives of the Biomedical Applications Unit are supported by the work of the Computational Chemistry Group. This team also supports the research activities of other groups of the Institute.
Other research interests of the Team focus at screening for candidate prognostic markers and/or therapeutic targets for several types of cancer, including skin, bladder, uterine and lung cancers. This is mainly achieved through the full elucidation of the role of crucial molecules in the mechanisms of carcinogenesis and the subsequent development of corresponding targeting strategies. The genes to be studied are selected based on the InstituteĘs previous experience, homology with well-known tumour suppressors or oncogenes, as well as on indicative bioinformatics and high-throughput methodsĘ results. In this context, the UnitĘs interests for the last decade have been mainly focused in the following fields:
Externally funded projects supportive of the R&D activities of the Unit (2005- ):
ENTER-01EP94: (2003-2007): "Role of MEF2 transcription factor in heart hypertrophy and cancer development". Coordinator: V. Zoumpourlis. (Budget 73.3 K€.), Participant: Antisel SA
Czech-Greek bilateral cooperation (2003-2005): "Role of the p53 oncosuppressor gene in androgen-independent prostate cancer". Coordinator: V. Zoumpourlis.(Budget 24 K€) Participant: Antisel SA
French-Greek bilateral cooperation (2004-2006): "Molecular mechanism for Androgen Receptor interaction with AP-1 in Prostate cancer". Coordinator: V. Zoumpourlis.(Budget 24 K€.) Participant: Antisel SA
USA - Greek bilateral cooperation (2005- 2008) «Molecular dissection of the mechanism of Rho kinase in susceptibility to cancer progression and metastasis». Coordinator: V. Zoumpourlis; (Budget 60K€.)
French-Greek bilateral cooperation (2004-06; GSRT): ''Molecular mechanism for Androgen Receptor interaction with AP-1 in Prostate cancer.'' Coordinator: Dr. V. Zoumpourlis. Unit budget: 23.3 K?.
EU project: ΤΟΚ (2007-2010) «Supramolecular chemistry and gene therapeutic potential of amine-amine-substituted cyclodextrin end-functionalized triazine dendrimers based on melamine" Contributor V. Zoumpourlis (budget 332 K€)
Peer-reviewed publications (2000 - ):
Progression of mouse skin carcinogenesis is associated with increased estrogen receptor alpha (ERα) levels and is repressed by a dominant negative form of ERα Dimitra Papaevangeliou, Stella logotheti, , Ioannis Christodoulou Katerina Pyrillou, John Michalopoulos, Vassilis G. Gorgoulis, Spiros Vlachopoulos and Vassilis Zoumpourlis. (Submitted 2011)
Global DNA hypomethylation-induced DNP73 transcriptional activation in non-small cell lung cancer. Alexandros Daskalos, Soultana Markopoulou, George Xenarianos, John R Cosney, Stella Logotheti, Vassilis Zoumpourlis. John K. Field, and Triantafyllos Liloglou, Cancer Lett. 300(1):79-86, 2011
Logotheti S, Michalopoulos I, Sideridou M, Daskalos A, Kossida S, Spandidos DA, Field JK, Vojtesek B, Liloglou T, Gorgoulis V, Zoumpourlis V. Sp1 binds to the external promoter of the p73 gene and induces the expression of TAp73gamma in lung cancer. FEBS J 277: 3014-3027, 2010
Rizos EN, Michalopoulou PG, Siafakas N, Stefanis N, Douzenis A, Rontos I, Laskos E, Kastania A, Zoumpourlis V, Lykouras L. Association of Serum Brain-Derived Neurotrophic Factor and Duration of Untreated Psychosis in First-Episode Patients with Schizophrenia. Neuropsychobiology 62: 87-90, 2010
Volanis D, Kadiyska T, Galanis A, Delakas D, Logotheti S, Zoumpourlis V. Environmental factors and genetic susceptibility promote urinary bladder cancer. Toxicol Lett. 193: 131-137, 2010
Vassilis G. Gorgoulis, Michalis Liontos, Katerina Niforou, Georgia Velimezi Konstantinos Vougas, Konstantinos Evangelou, Radek Vrtel, Alexandros Damalas, Panayiotis Kontovazenitis, Athanassios Kotsinas, Vassilis Zoumpourlis, George Th. Tsangaris, Christos Kittas, Doron Ginsberg, Thanos D. Halazonetis and Jiri Bartek3 Modulation of the E2F1-driven cancer cell fate by the DNA damage response machinery and potential novel E2F1 targets in osteosarcomas. Am J Pathol. 175:376-91, 2009.
7. Woodcock S, Rooney C, Liontos M, Connolly Y, Zoumpourlis V, Whetton A, Gorgoulis V and A Malliri. Src-induced disassembly of adherens junctions requires localised phosphorylation and degradation of Tiam-1. Molecular Cell 33, 1-15, 2009.
Copland J, Sheffield-Moore M, Koldzic Zivanovic C, Lamprou G, Tzortzatou-Stathopoulou F, Zoumpourlis V, Urban R, and Vlahopoulos S Sex steroids and their receptors at the crossroads between differentiation and neoplasia: a perspective for therapeutic intervention. Bioessay 31:629-41, 2009
Georgios Pampalakis, Evangelia Prosnikli, Theodora Agalioti, Antonia Vlahou, Vassilis Zoumpourlis and Georgia Sotiropoulou. A tumor protective role for human kallikrein 6 in breast cancer. Cancer Res 69: 3779-87, 2009.
Frydrych I, Mlejnek P, Dolezel P, Zoumpourlis V, Krumpochova P. The broad-spectrum caspase inhibitor Boc-Asp-CMK induces cell death in human leukaemia cells. Toxicol in Vitro 22: 1356-60, 2008
Spiros A. Vlahopoulos, Stella Logotheti, Dimitris Mikas, Athina Giarika, Vassilis Gorgoulis and Vassilis Zoumpourlis. The role of ATF-2 in oncogenesis. Bioessay 30: 314-327, 2008.
Bakas P, Liapis A, Vlahopoulos S, Giner M, Logotheti S, Creatsas G, Meligova AK, Alexis MN, Zoumpourlis V. Estrogen receptor alpha and beta in uterine fibroids: a basis for altered estrogen responsiveness. Fert. Ster. 1: 1-8, 2008
Liontos M, Koutsami M, Sideridou M, Evangelou K, Kletsas D, Levy B, Kotsinas A, Nahum O, Zoumpourlis V, Kouloukoussa M, Lygerou Z, Taraviras S, Kittas C, Bartkova J, Papavassiliou AG, Bartek J, Halazonetis TD, Gorgoulis VGDeregulated overexpression of hCdt1 and hCdc6 promotes malignant behavior. Cancer Research 67: 10899 -10909, 2007
Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva N, Vassiliou LV, Kolettas E, Niforou K, Zoumpourlis VC, Takaoka M, Nakagawa H, Tort F, Fugger K, Johansson F, Sehested M, Andersen CL, Dyrskjot L, Orntoft T, Lukas J, Kittas C, Helleday T, Halazonetis TD, Bartek J, Gorgoulis VG. Oncogene-induced senescense is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature 444: 633-637, 2006
Vlahopoulos S, Zimmer WE, Jenster G, Belaguli NS, Balk SP, Brinkmann AO, Lanz RB, Zoumpourlis V and Schwartz RJ. Recruitment of the androgen receptor via serum response factor facilitates expression of a myogenic gene. JBC 280: 7786-9, 2005
Papoutsi Z, Kassi E, Papaevangeliou D, Pratsinis H, Zoumpourlis V, Halabalaki M, Mitakou S, Kalofoutis A and P moutsatsou. Plant 2-Arylobenzofurans demonstrate a selective estrogen receptor modulator profile. Steroids 69: 727-734, 2004
Papassava P, Gorgoulis V, Papaevangeliou D, Vlahopoulos S, van Dam H, and Zoumpourlis V. Overexpression of Activated Transcription Factor 2 is required for tumor growth and progression in mouse skin tumors. Cancer Res 64: 8573-8584, 2004.
Vlachopoulos S and V Zoumpourlis. "JNK": a key modulator of intracellular signaling. Biochemistry (Moscow) 69: 844-854, 2004
Zoumpourlis V, Solakidi S., Papathoma A. and Papaevangeliou D. Genetic alterations in signal transduction pathways implicated in tumour progression during multistage mouse skin carcinogenesis. Carcinogenesis 24: 1159-1165, 2003.
Gorgoulis V, Zacharatos P, Mariatos G, Kletsas D, Zoumpourlis V, Kittas Ch, Ryan K and Papavassiliou A.. p53 activates ICAM-1 (CD54) expression in an NF-κB-independent manner. EMBO J 22: 1567-1578, 2003
Gazouli M, Kokotas S, Zoumpourlis V, Zacharatos P, Mariatos G, Kletsas D, Kittas Ch and Gorgoulis V. The complement inhibitor CD59 and the lymphocyte function-associated antigen-3 (LFA-3, CD58) genes possess functional binding sites for the p53 tumor suppressor protein. Anticancer Research 22: 4237-4242, 2002.
Katsanakis KD, Gorgoulis V, Papavassiliou A and Zoumpourlis V. The progression in the mouse skin carcinogenesis model is dependent on ERK1/2 signaling. Molecular Med 8: 624-637, 2002
Psichari E, Balmain A, Plows D, Zoumpourlis V and Pintzas A. High activity of serum response factor in the mesenchymal transition of epithelial tumor cells is regulated by Rho signaling. J Biol Chem. 277, 29490-29495, 2002.
Katsanakis K, Owen C, Zoumpourlis V. JNK and ERK signaling pathways in multistage mouse carcinogenesis: studies in the inhibition of signaling cascades as a means to understand their in vivo biological role. Anticancer Research 22, 755-760, 2002.
Plows D, Briassouli P, Owen C, Zoumpourlis V, Garrett M and A Pintzas. Ecdysone-inducible expression of oncogenic Ha-Ras in NIH 3T3 cells leads to transient nuclear localization of activated extracellular signal-regulated kinase regulated by mitogen-activated protein kinase phospatase-1. Biochem J 362, 305-315, 2002
Papathoma A, Zoumpourlis V, Balmain A and A Pintzas. Role of Matrix Metalloproteinase-9 in Progression of Mouse Skin Carcinogenesis. Molecular Carcinogenesis 31, 74-82, 2001.
Kotsinas A, Gorgoulis VG, Zacharatos P Mariatos G, Kokotas S, Liloglou T, Ikonomopoulos Zoumpourlis V, Kyroudi A, Field JK, Asimakopoulos PJ and Kittas Ch. Additional characterizaation of a hexanucleotide polymorphic site in the first intron of human H-ras gene: comparative study of its alterations in non-small cell lung carcinomas and sporadic invasive breast carcinomas. Cancer Genetics and Cytogenetics 126, 147-154, 2001
Gorgoulis VG, Zacharatos P Mariatos G, Liloglou T, Kokotas S, Kastrinakis N, Kotsinas A, Athanasiou A, Foukas P, Vogiatzi T, Zoumpourlis V, Kletsas D, Ikonomopoulos J, Asimakopoulos PJ, Rossidakis G, Kittas Ch and Field JK. Deregulated expression of c-mos in non-small cell lung carcinomas: Relationship with p53 status, genomic instability and tumor kinetics. Cancer Research 61, 538-549, 2001
Zoumpourlis Vassilis, Papassava Paraskevi, Linardopoulos Spyros, Gillespie David, Balmain Allan and Alexandros Pintzas. High levels of phosphorylated c-jun, Fra-1, Fra-2 and ATF-2 proteins correlate with malignant phenotypes in the multistage mouse skin carcinogenesis model. Oncogene 19, 4011-4021, 2000.
1. AP-1 transcription factors (mainly ATF-2) and steroid receptors in mouse skin carcinogenesis model
(The project has been supported by the USA - Greek bilateral cooperation (2005-2008) and by Marie Curie grant (Supragene, EU project, ΤΟΚ (2007-2010)).
1a. Investigation of the mechanisms of skin cancer progression The lab of Biomedical Applications Unit possesses a series of cell lines representing the distinct stages in mouse carcinogenesis, which include immortalized cells (C50), benign papillomas (cell lines P1, P6), locally invasive squamous carcinomas (B9, PDV, PDVC57) as well as aggressive spindle cell lines with metastatic potential in vivo (A5, D3, CarB, CarC). Importantly, the pairs B9:A5 and PDV:PDVC57 are useful for the investigation of alterations in genetic and signal transduction pathways during progression and epithelial-to-mesenchymal transition, since they represent the clonal expansion of cancer. The mouse skin carcinogenesis model, is an ideal experimental tool for the study of mechanisms of cancer progression (Fig 1). Research in this area is focusing on using this model to elucidate the role of Ras/JNK/ERK pathway and the AP-1 transcription factors in cancer progression and to devise ways of suppressing or even reversing the progression. Our findings suggest an involvement of AP-1 (c-Jun, Fra, ATF-2) and ERK1/2 in the development of biologically aggressive behavior of spindle mouse skin cells. Additionally we have shown that the use of dnATF-2 in targeting against wild type ATF-2 could block proliferation of cancer cells in skin tumours (Fig 2). We aim to generate optimum conditions for studying the role of ATF-2 in cancer progression and to form a basis for efficient use of recombinant ATF-2 in future translational studies (Zoumpourlis et al., Oncogene 19: 4011-4021, 2000; Katsanakis et. al.Anticancer Research 22, 755-760, 2002, Katsanakis et al., Molecular Med 8: 624-637, 2002; Zoumpourlis et. al. Carcinogenesis 24: 1159-1164, 2003; Papassava et al. Cancer Res 64: 8573-8584, 2004; Vlahopoulos et. al Bioessay 31:629-41, 2008)
Figure 1: Summary of key alterations regulating stage-to-stage transition during mouse skin carcinogenesis. A series of cell lines representing the three distinct stages in mouse skin carcinogenesis (initiation, promotion and progression) is shown (Zoumpourlis et. al. Carcinogenesis 24, 1159-1164, 2003).
Figure 2: Effect of dnATF-2 overexpression on in vivo tumorigenesis in BALB/c SCID mice (Papassava et. al Cancer Res 64, 8573-84, 2004, Vlahopoulos et. al Bioessay 31:629-41, 2008).
1b. Involvement of Estrogen Receptor alpha and beta in mouse skin carcinogenesis and in human uterine fibroids. Our findings suggest an involvement of ERα in the development of biologically aggressive behavior of spindle mouse skin cells, possibly through direct or indirect regulation of genes encoding for epithelial markers and cell adhesion properties. We also provide robust in vivo evidence that ERα-specific targeting can effectively suppress skin cancer progression (Papaevageliou et al Submitted, 2011).
Additionally we have shown that the levels of mRNA ERα and ERβ, as well as the total ER protein levels increased in leiomyomas compared with normal myometrium. However the estrogen receptor alpha-to-estrogen receptor beta expression ratio rather than the individual expression levels determines the fraction of DNA-binding homodimers of estrogen receptor alpha and possibly the growth potential of myomas (Bakas P et al Fert. Ster. 1: 1-8, 2008) [top]
2. The study of the role of p73 isoforms in human lung cancer (The project has been supported by the USA - Greek bilateral cooperation (2005-2008)and by the competitive GSRT grant ENTER 01EP94 (2003-2007)).
Lung cancer is one of the most prevalent and deadly types of cancer. Despite the progress that has been accomplished in the elucidation of the molecular mechanisms of carcinogenesis for the last 20 years, the number of lung cancer-related deaths has not been substantially reduced, mainly due to the failure of peatients to respond to current chemotherapy regimens. Hence, the identification of pivotal molecules of cancer pathogenesis that could be exploited as biological markers for early detection of the disease or/and as effective targets of molecular therapy is of critical importance.
p73, a p53 homologue, has been rarely found mutated in cancer and its effect in lung cancer is equivocal. p73 gives rise to a large number of isoforms. It has been suggested that p73 can produce isoforms with opposite activities, mainly the apoptotic TA and the antiapoptotic ΔΝ isoforms, emerging from the extrinsic P1 or the intrinsic P2 p73 promoter, respectively. In this respect, we have analyzed P1 and P2 promoters, in search of epigenetic and transcription factors that differentially regulate TA and ΔΝ isoform protein synthesis and, thus, determined the ultimate effect of p73 in lung cancer. We also identified the specific TAp73 or/and ΔΝp73 isoforms that are overexpressed in lung cancer. We found that P1 promoter is mainly deregulated by transcriptional events, whereas P2 promoter is deregulated by global-induced demethylation in non-small cell lung cancer.(Logotheti et al FEBS J 277: 3014-3027, 2010; Daskalos et al, Cancer Letters, 300; 79-86, 2011) [top]
3. The study of the role of Rho kinases in the progression and metastasis of cancer
(The project has been supported by the USA - Greek bilateral cooperation (2005-2008) and by the competitive GSRT grant ENTER 01EP94 (2003-2007)).
Rho proteins are members of the Ras superfamily and are homologous to the typical oncogene ras. Due to their documented role in the maintenance of the cell structure, the cell mobility and the cell adhesion-parameters that they are all related to the establishment of aggressive properties of cancer cells-they are implicated in invasive and metastatic cancer phenotypes. This project is focused in the study of: a) the changes in the expression levels of Rho protein kinases in the multistage cancer progression, b) the changes in the biological parameters and the oncogenic potential caused by specific knock-down of the Rho family member cdc42 and, c) the alterations of genomic profile in cancer cells which are related with Cd42 knock-down. For this study the mouse skin carcinogenesis model, numerous cell lines provided by the American Type Culture Collection, as well as an adequate number of primary lung- and bladder-derived tumours are recruited. The ultimate goal of this study will be the elucidation of the role of Rho kinases, as well as their evaluation as potential prognostic biomarkers in various cancer types.(Volanis et al. Toxicol Lett. 193: 131-137, 2010; Volanis et al submitted) [top]
4. Collaborative projects: the implication of kallikrein 6, Src oncoprotein, as well as the replication licensing factors hCdt1 and hCdc6 in malignancy.
4a. The role of human kallikrein-related peptidase 6 as an inhibitor of epithelial - to - mesenhymal transition in breast cancer.(Biomedical Applications Unit; Department of Pharmacy, University of Patras; Assoc. Prof. G Sotiropoulou).We investigated the mechanism(s) underlying the silencing of KLK6 gene in metastatic breast cancer and its putative implications for tumor progression. We found that KLK6 may play a protective role against tumor progression that is likely mediated by inhibition of epithelial-to-mesenchymal transition. (Pampalakis et al, Cancer Research 69: 3779-87, 2009) [top]
4b. The role of SRC oncoprotein in the phosphorylation and degradation of the rac activator tiam1 and their effects in epithelial to mesenchymal transition (EMT). (Biomedical Applications Unit; Cell Signalling Group, Cancer Research UK Paterson Institute for Cancer Research, University of Manchester, UK, Dr Agelliki Malliri). We showed that Tiam1 phosphorylation is needed for vSrc-induced disruption of adherens junctions (AJs) and migration, and we presented evidence that this mechanism acts in a range of human malignancies. (Woodcock et al Molecular Cell 33, 1-15, 2009) [top]
4c. The implication of hCdt1 and hCdc6 in the progression of malignancy. (Biomedical Applications Unit; Medical school of Athens, Professor Vassilis Gorgoulis). Abnormal accumulation of hCdt1 and hCdc6 proteins occurred early from the stage of dysplasia. Overexpression of hCdt1 and hCdc6 promoted rereplication and generated a DNA damage response, which activated the antitumor barriers of senescence and apoptosis. In addition, stable expression of hCdc6 and hCdt1 in premalignant papilloma cells led to transformation of the cells that produced tumors upon injection into nude mice depicting the oncogenic potential of their deregulation. (Liontos et al Cancer Research 67: 10899 -10909, 2007; Nature 444: 633-637, 2006) [top]