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December 5-7 2019 Royal Olympic Hotel Athens Greece
Symposium on Advances in Cancer Immunology and Immunotherapy
The event will be awarded by Panhellenic
Medical Association with 18
SCIENTI F I CP R OG R A M
Hellenic Society of Immuno-Oncology
Under the Auspices of
Scientific Ι Cultural Events & Publications Τ: +30 210 7240039, +30 2810 222155F: +30 210 7240139, +30 2810 222156 E: [email protected] www.scep.gr
Symposium on Advances in Cancer Immunology and ImmunotherapySymposium on Advances in Cancer Immunology and Immunotherapy
Welcome LetterDear Colleagues,
We are delighted to welcome you to the 5th Symposium: Advances in Cancer Immunology and Immuno-
therapy, that is being held on December 5-7, 2019 at the Royal Olympic Hotel, Athens, Greece.
The meeting aims to bring together many of the leading researchers and clinicians in Europe and the US to
report on most recent advances made in cancer immunology and immunotherapy. Our main objectives is to
gather, share and exchange experiences and ideas; translate and extend this knowledge to the clinic; promote
interactions between speakers and participants and encourage stimulating discussions; provide network
opportunities and stimulate new collaborations. This year, we have introduced a new element, as most young
researchers that will participate in the poster sessions will be additionally given the opportunity to present
the highlights of their work as a short oral presentation. The Scientific Committee will select the three best
presentations for awards.
On behalf of the Organizing and Scientific Committees, we are very pleased to have you amongst us.
We hope to enjoy the Symposium.
The Organizing Committee Chairs
Athanasios Kotsakis, MD, PhD
Associate Professor of Medical Oncology,Director of Dpt of Medical Oncology,
University Hospital of Larissa,Thessaly, Greece
Ourania Tsitsilonis, MD, PhD
Associate Professor of Immunology,Department of Biology,National & Kapodistrian
University of Athens, Greece
Constantin N. Baxevanis, PhD
Scientific Director,Cancer Immunology andImmunotherapy Center
“Agios Savvas” Hospital, Athens, Greece
Symposium on Advances in Cancer Immunology and ImmunotherapySymposium on Advances in Cancer Immunology and Immunotherapy
Organizing CommitteeAthanasios Kotsakis
Constantin N. Baxevanis
Scientific CommitteeC. N. Baxevanis
E. K. Vetsika
Symposium on Advances in Cancer Immunology and ImmunotherapySymposium on Advances in Cancer Immunology and Immunotherapy
General infoOrganized by Hellenic Society of Immuno-Oncology (HeSIO) G. Theologou 5, Τ.Κ. 11474, Athens, Greece, www.hellenic-immunooncology.gr
Under the Auspices of
Dates December 5th - 7th, 2019
Symposium Venue Royal Olympic Hotel, Athens 28-34, Athanasiou Diakou Str., 117 43, Athens, GREECE Tel.: +30 210 92.88.400, Fax: +30 210 92.33.317 E-mail: [email protected]
Registrations Registration Fee
• Undergraduate students and postgraduate students - € • PhD students - € • Residents - € • Post-docs and MD 200€ • The Orginizing Committee has decided to give the opportunity to 20 young doctors to participate at the Symposium with Free Registration Fee.
Certificate of AttendanceBased on the latest circular of the National Drug Organization the Symposium is required to use an attendance tracking system. All registered participants will receive name badges, which they are kindly requested to wear at all times. Each badge will have a barcode for monitoring the hours of attendance. By the end of the Symposium a certificate will be given to those who have attended at least 60% of the total hours of the Scientific Program
The event has been awarded by Panhellenic Medical Association (PIS) with 18 CME - CPD credits
Official Language English is the official language of the Symposium
S c i e n t i f i c Ι C u l t u r a l E v e n t s a n d P u b l i c a t i o n s T +30 210 7240039, +30 2810 222155 F +30 210 7240139, +30 2810 222156 E [email protected] www.scep.gr
Thursday, December 5th, 201915.00-15.15 W e l c o m e - R e g i s t r a t i o n s
15.15-16.45 Session 1: Novel Immunotherapies for Cancer Chairs: V. Georgoulias, A. Kotsakis 15.15-15.45 The cystine/glutamate antiporter xCT: one tumor antigen, multiple functions F. Cavallo 15.45-16.15 CD4+ T cells as an emerging solution to treat solid tumors EM. Inderberg 16.15-16.45 Genetically Modified T Cells for the Immunotherapy of Cancer M. Nishimura 16.45-17.15 C o f f e e b r e a k
17.15-19.30 Session 2: Aging and cellular senescence in the era of cancer immunotherapy Chairs: O. Tsitsilonis, I. Trougakos 17.15-17.45 Harnessing NK cells for cancer immunotherapy: impact of NK cell immunosenescence R. Solana 17.45-18.15 Immunotherapy of cancer: triumphs and challenges, and the impact of immunosenescence in humans - is aging always a bad thing? G. Pawelec 18.15-18.45 Interplay between cellular senescence and cancer D. Kletsas
Plenary lecture Chairs: C. Baxevanis, G. Pawelec 18.45-19.30 The role of microRNA in cancer B. Seliger19.30 D i n n e r
Friday, December 6th, 201909.30-11.00 Session 3: Tumor immune resistance and escape Chairs: F. Cavallo, C. Baxevanis 09.30-10.00 Secondary immune resistance mechanisms induced by therapeutic cancer vaccines which prevent tumor regression and foster recurrences S.H. Van der Burg 10.00-10.30 Understanding how the immune system prevents tumor escape A. Madrigal 10.30-11.00 Roadblocks to cancer immunotherapy J. Wischhusen 11.00-11.30 C o f f e e b r e a k
11.30-13.00 Session 4: Tumor regression vs tumor progression: balancing T and NK cells vs Neutrophils and MDSC Chairs: A. Madrigal, R. Solana 11.30-12.00 A complex interplay between dendritic cells, NK cells and T cells governs adaptive immune responses in tumor microenvironments M. Tsoumakidou 12.00-12.30 Nuclear factor-kappa B-based tumor-host interactions in malignant pleural disease G. Stathopoulos 12.30-13.00 Neutrophils and MDSC as mediators of immune suppression and drivers of tumor progression S. Brandau13.00-14.30 L u n c h
14.30-16.20 Short Oral Presentations (I) Chairs: V. Georgoulias, O. Tsitsilonis 14.30-14.40 N-BROMOTAURINE AND ITS STABlE ANAlOGUE MOlECUlE (BROMAMINE T-BAT) ExHIBIT A THERAPEUTIC EFFECT AGAINST CANCER AND INFLAMMATION IN VITRO AND IN VIVO S. Baliou 14.40-14.50 lUNG TUMOR IMMUNITy IS SUSTAINED lOCAlly By MHC-II ANTIGEN PRESENTING FIBROBLASTS ExPRESSING ATyPICAL CO-STIMUlATORy MOlECUlES K. Goudevenou 14.50-15.00 MAGNETIC NANOPARTICLES FOR IMMUNOGENIC CELL DEATH AND MOBILIzATION OF DENDRITIC CELLS K. Goudevenou 15.00-15.10 UlTRA-HyPOFRACTIONATED RE-IRRADIATION wITH ANTI-PD-1 IMMUNOTHERAPy FOR PATIENTS wITH RECURRENT OR RADIO-RESISTANT SqUAMOUS CEll HEAD AND NECK CANCER (SqCHNC) M. Koukourakis 15.10-15.20 ExPRESSION lEvElS OF PD-l1 IN HUMAN PROSTATE CANCER CEll lINES UNDER HyPOxIC AND ACIDIC CONDITIONS M. Koukourakis 15.20-15.30 PRECLINICAL STUDIES OF GLyCOVAxGM3 VACCINE: ROLE OF ADAPTIVE AND INNATE IMMUNE CELLS MEDIATING THE ANTITUMOR EFFECT M. Labrada Mon 15.30-15.40 IDENTIFICATION OF IMMUNE MODULATORy MICRORNAS TARGETING THE PEPTIDE TRANSPORTER TAP1 IN MELANOMA M. Lazaridou 15.40-15.50 CONCURRENT ExPOSURE TO HDAC-INHIBITORS AND TLR7 AGONISTS INDUCES APOPTOSIS IN OVARIAN CANCER CELLS VIA TRANSCRIPTIONAL ACTIVATION OF HUMAN ENDOGENOUS RETROVIRUSES V1/V2 S. Sahitya 15.50-16.00 THE PROGNOSTIC SIGNIFICANCE OF HlA-A*02:01 & HlA-A*24:02 ALLELES IN PROSTATE CANCER S. Stokidis 16.00-16.10 ERF ACTS AS A POTENTIAL POSITIVE EFFECTOR DURING T LyMPHOCyTES MATURATION IN THyMUS S. Tsiomita 16.10-16.20 INFLAMMATION RELATED MIRNAS AND BONE MARROW INFILTRATION IN MyElODySPlASTIC SyNDROME (MDS) RISK DETERMINATION AND THERAPy RESPONSE C. Vaxevanis16.20-17.00 C o f f e e b r e a k
17.00-18.30 Session 5: Immunomodulation and novel immune checkpoints Chairs: G. Adema, B. Seliger 17:00-17:30 Opening a new therapeutic window for human bladder cancer D. Stravopodis 17.30-18.00 Immunomodulatory properties of chitinase-like proteins in tumor J. Kzhyshkowska 18.00-18.30 Discovery and validation of novel immune checkpoints for cancer immunotherapy in humanized mice G. Marοdon
18.30-19.30 Plenary lectures Chairs: D. Stravopodis, G. Kyrgias 18.30-19.00 Lipids as Immune Modulators of Myeloid Cells in Cancer G. Adema 19.00-19.30 Tumor Microenvironment and response to Radiotherapy and Immunotherapy MI. Koukourakis
Saturday, December 7th, 201909.30-11.30 Session 6: The tumor microenvironment and immune regulation Chairs: Α. Kotsakis, A. Koutsopoulos 09.30-10.00 Immune cell infiltration in breast cancer A. Giatromanolaki 10.00-10.30 Targeting TGF-beta production by GARP-expressing Tregs for cancer immunotherapy S. Lucas 10.30-11.00 Dynamic interactions within the tumor microenvironment P. Foukas 11.00-11.30 The targeting of the immune suppressive microenvironment with anti-regulatory T cells E. Martinenaite 11.30-12.00 C o f f e e b r e a k
12.00-13.30 Session 7: Immunotherapies (I) Chairs: A. Kotsakis, V. Georgoulias 12.00-12.30 Cell-based Immunotherapy of solid tumors J. Bartůňková 12.30-13.00 Immunotherapy in first-line treatment of Non-Small Cell Lung Cancer E. Kontopodis 13.00-13.30 Update of the role of immunotherapy in melanoma Α. Laskarakis13.30-15.30 L u n c h
15.30-16.00 Short Oral Presentations (II) Chairs: O. Tsitsilonis, G. Pawelec Ιs there any future for Cancer vaccines? K. Kosmatopoulos
16.00-17.30 Session 8: Immunotherapies (II) Chairs: V. Georgoulias, A. Koumarianou 16.00-16.30 A novel biomarker and a new target for the immunotherapy of early stage ovarian cancer B. Guinn 16.30-17.00 The emerging role of combinatorial regimens in renal cancer* K. Koutsoukos *sponsored by 17.00-17.30 latest data for immunotherapy in urothelial cancer* F. Koinis *sponsored by 17.30-18.15 C o f f e e b r e a k
18.15-19.15 Session 9: Immunotherapies (III) Chairs: C. Baxevanis, J. Bartůňková 18.15-18.45 Adverse events from immunotherapy: Are they so rare? A. Koumarianou 18.45-19.15 Hyperprogressive disease during immunotherapy N. Pistalmatzian19.15 A w a r d s - C l o s i n g R e m a r k s
Sponsored Lectures16.00-17.30 Session 8: Immunotherapies (II) Chairs: V. Georgoulias, A. Koumarianou
16.30-17.00 The emerging role of combinatorial regimens in renal cancer* K. Koutsoukos *sponsored by
17.00-17.30 latest data for immunotherapy in urothelial cancer* F. Koinis *sponsored by
AbstractsCARBONIC ANHYDRASE 9 (CA9) AND IMMUNE TOLERANCE IN NON-SMALL CELL LUNG CANCER (NSCLC)A. Giatromanolaki , I.M. Koukourakis , T. Alexiadis , A.L. Harris , A.H. Banham , M.I. Koukourakis  Department of Pathology and Department of Radiotherapy/Oncology, Democritus University of Thrace, Alexandroupolis, Greece;  National and Kapodistrian University of Athens, Greece;  Nuffield Division of Clinical Laboratory Sciences and Cancer Research UK Molecular Oncology Laboratories, University of Oxford, Oxford, UK
Introduction: Immunotherapy with immune checkpoint inhibitors has revolutionized the therapy of cancer. Anti-PD-1/PD-L1 monoclonal antibodies provide prolongation of life and response rates to immunotherapy about 25%. Intratumoral hypoxia and acidosis are also implicated in tumor evasion from immune surveillance. Glutamine metabolism feeds the Krebs cycle with gluta-mate and a-ketoglutarate, sustaining oxidative metabolism under hypoxia, producing CO2. CO2 is hydrated by the transmembrane protein carbonic anhydrase 9 (CA9) and converted to bicarbonate and protons that acidify the extracellular milieu. Low pH mark-edly suppresses the proliferation and cytotoxic activity of CD8+ and natural killer lymphocytes.
Methods: We examined whether CA9 is also involved in intratumoral immunosuppression pathways, in a series of 98 non-small cell lung carcinomas (NSCLC) treated with surgery. The expression of CA9 and programmed-death ligand PD-L1 by cancer cells was examined in parallel with FOxP3 expression by tumor infiltrating lymphocytes (TIls), a marker of regulatory immunosup-pressive CD4+ lymphocyte subset.
Results and Conclusions: CA9 was not linked with PD-L1 expression or the density of TILs in the neoplastic stroma. A direct sig-nificant association of CA9 expression with the extent of FOxP3+ TIl density was noted (p=0.008). Patients with high CA9 expres-sion and low TIl-score had significantly poorer survival compared to all other groups (p
LUNG TUMOR IMMUNITY IS SUSTAINED LOCALLY BY MHC-II ANTIGEN PRESENTING FIBROBLASTS EXPRESSING ATYPICAL CO-STIMULATORY MOLECULES *1D. Kerdidani, *1K. Goudevenou, 1M. Aerakis, 1K. Verrou, 1C. Tzaferis, 1A. Prados, 2G. Vamvakaris, 3V. Kaniaris, 4K. Vachlas, 4K. Potaris, 5T. Koutsopoulos, 1M. Tsoumakidou1 BSRC Alexander Fleming, Department of Immunology, Vari, Greece, 2 Pathology Department, Athens Chest Hospital “Sotiria”, Ath-ens, Greece, 3 ICU First Respiratory, University of Athens, Sotiria General Hospital, Athens, Greece, 4 Department of Thoracic Sur-gery, Sotiria General Hospital, Athens, Greece, 5 Department of Pathology, University General Hospital of Heraklion, Crete, Greece
*These authors contributed equally
The immunological importance of local T cell activation for effective tumor immunity is incompletely understood. Current paradigm states that the population of tumor-specific T cells is sustained at tumor sites by intratumoral cDCs. However, convincing data to support the leading role of intratumoral cDCs as the main local antigen-presenting cells (APCs) is lacking. Non-haematopoietic APCs can be induced to express MHCII, but due to the lack of co-stimulatory molecule co-expression it has long been postulated that they suppress T cell responses. Very recent works revealed unrecognized functions of intestinal epithelial cells in priming T cells and of fibroblastic reticular cells (FRCs) in supporting CD4+ and CD8+ T cells, questioning typical thinking of atypical APCs as tolerogenic cells. A new subset of MHCII-expressing cancer-associated fibroblasts (apCAFs) in pancreatic adenocarcinoma was recently discovered. It was postulated that apCAFs induce CD4+ differentiation into Tregs. In this study, we have found that up to one third of CAFs in lung carcinomas can be classified as apCAFs. Using several cytokine-targeting mouse strains and 3D cul-tures, we show the requirement of strong cytokine milieu for CAF activation to apCAFs. Unexpectedly, fibroblast-specific targeted ablation of MHC Class II decreased lung tumor burden and tumor-infiltrating CD4+ T cell numbers, impacted their transcriptomic and phenotypic profile, accompanied by a decrease in CD8+ T cells. Primary human CAFs directly presented MHCII-peptide com-plexes to activate autologous intratumoral CD4+ T cell. Unbiased single cell RNA-seq unveiled environmental regulation of human apCAFs and unconventional T cell priming potential due to expression of atypical co-stimulatory molecules. These results show that lung fibroblasts are key APCs sustaining locally anti-tumor T cell responses and need to be considered when designing im-munotherapeutic strategies to increase intratumoral cancer antigen-specific T cells.
N-BROMOTAURINE AND ITS STABLE ANALOGUE MOLECULE (BROMAMINE T-BAT) EXHIBIT A THERAPEUTIC EFFECT AGAINST CANCER AND INFLAMMATION IN VITRO AND IN VIVO Stella Baliou1**, Markus Nagl2, Antony M. Kyriakopoulos3, and Vassilis Zoumpourlis1*1. Biomedical Applications Unit, Institute of Biological Research and Biotechnology, National Hellenic Research Foundation, 48 Vas. Constantinou Ave., 116 35 Athens, Greece, 2. Division of Hygiene and Medical Microbiology, Medical University of Innsbruck, Schöpfstr. 41, 6020 Innsbruck, Austria, 3. NASCO AD Biotechnology Laboratory, 11 Sachtouri str., 18536, Pireas, Greece
*Corresponding author: e-mail: [email protected]
Introduction: N-Bromotaurine is a physiological molecule which arises from the neutrophil -myeloperoxidase halide system of metabolism during inflammation and constitutes the reaction product of Taurine (Tau) with hypobromous acid (HOBr). N-Bromo-taurine is a potent oxidant molecule, which exerts a complex anti-inflammatory action. However, due to the poor stability of the N-Bromotaurine molecule, an analogue molecule, named Βromamine T (BAT), was subsequently used. Methods: Cell prolifera-tion experiments, colony formation assays, staining with CM-H2DCFDA dye, flow cytometry, Real-time PCR, immunofluorescence assay and western blot experiments highlighted the potential anti-cancer and anti-inflammatory effect of molecules. Α xenograft assay and an air-pouch model of inflammation induced by lPS constituted the in vivo experiments, delineating the functional significance of molecules. Results/Discussion: The tumor-suppressive effect of N-Bromotaurine was observed in various can-cer cell types, including skin cancer. In the case of skin cancer, we showed that N-Bromotaurine can bypass the glucocorticoid receptor (GR)-resistance of cancer cells when used in combination with cis-platin. Cancer cell proliferation was suppressed with the use of BAT in breast, cervical, and colorectal cancer, while the anti-proliferative effect of BAT appeared to be superior to that elicited by taurine. In addition, flow cytometry and western blot experiments highlighted the intrinsic apoptotic pathway used by cancer cells following exposure to BAT or Tau treatment alone. Additional experiments proved that BAT or Tau treatment alone triggered ROS accumulation in cancer cells, so that distinct members of the MAPK family were induced according to each treat-ment. As a result, BAT or Tau treatment alone induced oxidative burst, which in turn transduced signals to mediate the phosphory-lation of stress-related MAPK kinases, thereby eliciting pro-apoptotic, DNA damage and autophagic signals in colon cancer cells. In conditions of acute inflammation, BAT and Tau exerted anti-inflammatory properties by suppressing mRNA expression levels
of cytokines in both LPS-induced macrophages and in LPS-induced mice with air pouch formation. More importantly, xenograft experiments showed that tumor formation was impaired after treatment of immunodeficient mice with BAT or Tau. Our study has substantiated that BAT is an emerging anti-proliferative agent with favorable efficacy.
MATERNAL IMMUNIZATION AGAINST ALK: A NEW WEAPON AGAINST NEUROBLASTOMA Giuseppina Barutello1, Federica Riccardo1, Claudia Voena2, Roberto Chiarle2,3 and Federica Cavallo1 1 Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy, 2 Department of Molecular Biotechnology and Health Sciences, Center for Experimental Research and Medical Studies, University of Torino, Torino, 10126, Italy, 3 Children’s Hospital Boston, Boston MA, USA
Background: Neuroblastoma (NB) is the most common extracranial solid tumor in infancy. Some germline and somatic muta-tionsassociated with NB have been identified as activating mutations of the anaplastic lymphoma kinase (AlK) oncogene. Because of the nature of NB, which can occur in the early post-natal life or even during fetal life, we evaluated the efficacy of a DNA vaccine against AlK in a spontaneous preclinical model of NB, harboring AlKF1174l mutation in association with MyCN amplification (AlKF1174l/MyCN mice) by means of maternal immunization (MI).
Methods: MyCN transgenic females were immunized against ALK by DNA vaccination by using a plasmid coding for the extracel-lular and trans-membrane domains of human ALK (ALK-ECTM) followed by electroporation, and then they were mated with an AlKF1174ltransgenic male. In AlKF1174l/MyCN offspring, the presence of abdominal, cervical and paraspinal tumors has been evaluated and quantified by Magnetic Resonance Imaging. The humoral immune response induced against AlK in the mothers and their offspring, as well as the presence of immune complexes containing AlK, have been evaluated by ElISA. AlK expression in tumor tissue was assessed by Western blot.
Results: Pre-birth immunization against ALK leads to an extended survival time and to a lower tumor growth kinetic in ALK-F1174l/MyCN offspring born from AlK-ECTM-vaccinated mothers (AlK-ECTM offspring) as compared to controls born from control empty vectorvaccinated mothers. Maternally derived anti-ALK antibodies were successfully transferred from mothers to newborns. Moreover, anti-AlK IgM were found in the sera of five- and six-week old AlK-ECTM offspring, suggesting the induc-tion of the pups’ own immune response against AlK. This effect could be due to the breast milk-mediated transfer of immune-complexes containing AlK, found in the milk of vaccinated mothers and in their offspring sera. Finally, MI against AlK induces a decrease in AlK expression in AlK-ECTM offspring’ tumor tissue.
Conclusions: Overall, these results indicate that MI against ALK induces an active immunization against this oncoantigen in the offspring, impairing tumor development and enhancing survival time in a preclinical model of NB.
ERF ACTS AS A POTENTIAL POSITIVE EFFECTOR DURING T LYMPHOCYTES MATURATION IN THYMUSSpyridoula Tsiomita1, George Mavrothalassitis2,3, George Thyphronitis11 Department of Biological Applications and Technology, University of Ioannina, Ioannina, Greece, 2 IMBB, FORTH, Heraklion, Crete, Greece and 3 Medical School, University of Crete, Heraklion, Crete, Greece
BACKGROUND: The maturation of thymocytes in the thymus is a complex process of critical importance for the immune response and the immune tolerance. Maturation proceeds in successive stages and it is regulated by an extensive network of transcriptional factors and thymic signals. Having completed thymopoiesis and cell lineage commitment, thymocytes leave the thymus as naïve CD4+ or CD8+ T lymphocytes expressing functional T cell receptor and migrate to peripheral lymphoid organs.
ERF (ETS2 Repressor Factor) is a ubiquitously expressed transcriptional repressor of the ETS family that is regulated by subcel-lular localization as a result of Erk-dependent phosphorylation. It is well established that Erk1/2 plays an important role in T cell maturation and differentiation, however, its effectors are not well known.
METHODS: In the present research, we studied the role of Erf, in vivo, in thymocyte subpopulations during murine T cell matura-tion, using Erf floxed mice to produce thymus-specific ERF knockouts.
RESULTS: we observed statistically significant decreased numbers of CD4+ and TCRβ subpopulation and statistically significant elevated numbers of DP (CD4+CD8+) cells in the thymus of Erf deficient mice. Thus, our findings suggest that Erf is involved in T cell maturation at the DP stage, acting as a positive regulator during CD4 lineage commitment. Investigating the mechanism of Erf function, we focused on the expression of critical genes during lineage commitment and found that Gata3, ThPOK and Socs1 ex-
pression was statistically significant decreased in Erf deficient mice. Since those genes induce CD4 commitment, our findings are consistent with the hypothesis that Erf is a positive regulator of helper lineage commitment. Taking into consideration that Gata3 directly induces ThPOK expression and ThPOK regulates positively Socs1 expression, we propose that Erf acts as an upstream direct or indirect activator of Gata3.
CONCLUSIONS: Overall, our results show, for the first time, the possible involvement of ERF in T cell biology and can be the trig-ger for further research to understand the mechanisms underlying thymocyte maturation, central tolerance and anergy, leading to novel therapeutic strategies for cancer and autoimmune diseases.
IDENTIFICATION OF IMMUNE MODULATORY MICRORNAS TARGETING THE PEPTIDE TRANSPORTER TAP1 IN MELANOMA Maria - Filothei Lazaridou1, Chiara Massa1, Diana Handke1, Michael Friedrich1, Anja Mueller1, Reinhard Dummer2, Peter Koelblinger3, Barbara Seliger1 1Martin Luther University Halle-Wittenberg, Institute of Medical Immunology, Halle, Germany, 2University Hospital Zurich, Clinic for Dermatology, Zurich, Switzerland and 3Department of Dermatology, Paracelsus Medical University, Salzburg, Austria
Background Reduced HLA class I (HLA-I) surface expression can be mediated by impaired expression of antigen processing and presentation machinery (APM) components due to epigenetic, transcriptional and/or posttranscriptional processes. Since a discordant mRNA and protein expression pattern of HLA-I APM components, including the transporter associated with antigen processing 1 (TAP1), has been observed in melanoma, a posttranscriptional control caused by immune modulatory microRNAs (im-miRs) was sug-gested. Thus, the aim of this study is to identify and functionally characterize im-miRs directed against TAP1 in melanoma.
Methods In silico analysis as well as the miTRAP assay in combination with small RNA sequencing led to the identification of candidate miRs targeting the TAP1 3’ untranslated region (UTR). Their specific binding was evaluated by the dual luciferase reporter assay. Furthermore, the miR and APM component expression levels were determined by qPCR in both melanoma cell lines and primary melanoma lesions. Additionally, mimics or inhibitors of TAP1 directed candidate miRs were transfected into cells to modulate TAP1 expression. TAP1 protein expression levels were determined by western blot, while HlA-I surface expression by flow cy-tometry. NK cell- and/or CTL-mediated cytotoxicity was determined by the CD107 degranulation assay. Immunohistochemical analyses were performed on formalin-fixed paraffin-embedded (FFPE) samples of primary melanoma patients using respective antibodies.
Results Based on RNA sequencing data of miTRAP eluates, 211 miRs were enriched in miTRAP eluates of the TAP1 3’-UTR, which are in line with the 237 in silico predicted miRs using three bioinformatics tools. The dual luciferase reporter assay confirmed the specific binding of selected candidates, e.g. miR-26b-5p and miR-21-3p, to the TAP1 3’-UTR. Overexpression of miR-26b-5p and miR-21-3p reduced not only TAP1 mRNA and protein expression levels in selected melanoma cell lines by 35% and 30%, respectively, which was accompanied by a decreased HlA-I surface expression. The selective suppression of miR-26b-5p and miR-21-3p restored TAP1 along with HLA-I surface expression levels. The inverse correlation of TAP1, HLA-I with miR expression levels in melanoma cell lines was also found in primary melanoma lesions and further correlated with the frequency of CD8+ T cells.
Conclusions The data showed a miR-mediated downregulation of TAP1 and consequently a decreased HLA-I surface expression in melanoma cell lines and lesions suggesting the importance of the posttranscriptional control of HLA-I APM components, which might have a prognostic impact and might also predict the response to immunotherapies.
MAGNETIC NANOPARTICLES FOR IMMUNOGENIC CELL DEATH AND MOBILIZATION OF DENDRITIC CELLS1K. Goudevenou, 1E. Aerakis, 1D. Kerdidani, 2,3M. Theodosiou, 2,3M. Krokidis, 2,3E.K. Efthimiadou and 1M. Tsoumakidou1 Institute of Bioinnovation, BSRC Alexander Fleming, Vari, Greece, 2 Institute of Nanoscience and Nanotechnology, N.C.S.R. Demokritos, Athens, Greece and 3 Department of Chemistry, National and Kapodistrian University of Athens, Greece
Background. we have generated innovative magnetic nano-flowers (mNFls) for hyperthermia induction and cancer cell death at low concentrations. Immunogenicity of cell death depends on dendritic cells (DCs) but is hampered by DC exclusion and/or sup-
pression at tumor sites. Chemokines that could be used to attract DCs cannot be easily directed at tumor sites and diffuse quickly when injected in situ. Immune adjuvants become poorly immunogenic and toxic when administered systematically and in high dosages. Αim. We aimed to interrogate i) the immunogenicity of mNFL-induced cancer cell death and ii) the biological activity of DC targeting chemokines encapsulated in magnetic nanoparticles. Our ultimate goal is to develop an innovative immunotherapeu-tic approach based on the combination of magnetic hyperthermia with DC-targeting magnetic nanoparticles. Methods. mNFLs were administered i.v. in lung tumor-bearing mice. In one set of experiments mice were exposed to alternate magnetic field (hyperthermia) or sham. In another set of experiments magnetic nanocarriers of CCl4 or naked CCl4 were intratumorally injected into flank tumor-bearing mice. Tumors were lysed and analyzed by FACS. Results. Albeit there were signs of T cell responses (increased granzymeB, low T regulatory cells), hyperthermia alone did not increase CD8+ T cell infiltration and failed to rescue T cells from exhaustion, associated with poor mobilization (numbers, phenotype) of DCs. CCl4-containing magnetic nanocar-rriers effectively mobilized DCs and T cells at tumors. Conclusion. There is a limited immunotherapeutic potential of magnetic hyperthermia as stand-alone treatment, which points to the need for combinatorial treatment approaches with agents that target dendritic cells at tumors.
PRECLINICAL STUDIES OF GLYCOVAXGM3 VACCINE: ROLE OF ADAPTIVE AND INNATE IMMUNE CELLS MEDIATING THE ANTITUMOR EFFECTMayrel Labrada Mon1, Giselle Hevia Hernández1, Marilyn Clavell Pérez1, Luis E. Fernández Molina21Immunopotenciation and Biomarkers Department, Immunology and Immunotherapy Division, Molecular Immunology Institute (INIM), Center of Molecular Immunology (CIM), Havana, Cuba. 2Innovation Division, INIM, CIM, Havana, Cuba
Background: The GlycoVaxGM3 vaccine is a nanoparticulated product obtained through the insertion of GM3(Neu5Gc) ganglioside into the outer membrane protein complex of N. meningitides. The vaccine is currently studied in late stage clinical trials in breast cancer and melanoma patients, but mechanisms involved in the GlycovaxGM3’s antitumor effect are insufficiently understood. Here we have addressed the role of adaptive and innate immune cells in mediating the protective effect of the vaccine. Methods: To this aim we selected the 3LL-D122 Lewis lung spontaneous metastasis model, where the increase in GM3(Neu5Gc) expres-sion, resembles human cancer behavior, making it a relevant tumor model for studying GM3(Neu5Gc)-based vaccination. Results and Conclusions: Inoculation of the vaccine in tumor bearing mice induced anti-metastatic effect. The immunization of tumor bearing mice with GlycoVaxGM3 consistently induced anti-GM3(Neu5Gc) IgM and IgG Ab. Regardless the T-independent nature of GM3(Neu5Gc) ganglioside as antigen, the antimetastatic effect of GlycovaxGM3 was dependent of CD4+ and CD8+ T cells. In a further step we found that the vaccine was able to promote the increase, maturation and cytokine secretion of conventional dendritic cells. On the other hand the infiltration of NK1.1+CD3- and NK1.1+CD3+ cells in lungs of vaccinated mice was significantly increased, compared with the presence of these cells in control animal lungs. In the same way GlycoVaxGM3 mobilized acquired immunity effector cells into the lungs of vaccinated tumor bearing mice. leukocyte infiltration in lungs of tumor bearing mice correlates with vaccine induced inhibition of lung metastization. Finally, using the C26GM tumor model, it was demonstrated that GlycovaxGM3 treatment decreases the suppressive capacity of myeloid derived suppressor cells (MDSC) generated by the C26GM tumor model.
THE PROGNOSTIC SIGNIFICANCE OF HLA-A*02:01 & HLA-A*24:02 ALLELES IN PROSTATE CANCERSavvas Stokidis1*, Sotirios P. Fortis1*, Paraskevi Kogionou1, Marina I. Konstantellou1, Theodoros Anagnostou2, Sonia A. Perez1, Constantin N. Baxevanis1 * Equally contributed as first authors1 Cancer Immunology and Immunotherapy Center, Saint Savas Cancer Hospital, Athens, Greece, 2 Department of Urology, Saint Savas Cancer Hospital, Athens, Greece
BACKROUND: The major role of the immune system in cancer development and evolution is established. The human leukocyte antigen (HLA) complex is implicated in the manifestation of many diseases, also including cancer. As HLA-alleles are essential for an effective antitumor response, their expression could serve as prognostic/predictive biomarkers. Herein, we identified HlA-alleles as novel biomarkers, possibly contributing to a more accurate prognostic evaluation of prostate cancer (PCa).
METHODS: In this combined retrospective/prospective study, we enrolled PCa patients initially diagnosed between 1998-2018 with localized (lPCa, n=52) or metastatic (MPCa, n=53) disease. All patients received standard medical treatment upon diagnosis
and follow-up data, either retrospective or prospective, were recorded. Time to biochemical recurrence (BCR), metastasis (M), castrate resistance (CR) were evaluated. Correlations were also made with patient overall survival (OS). Patients were genotyped for the HLA- A locus.
RESULTS: Patients were stratified by HlA-A*02:01 & HlA-A*24:02 status. The Kaplan-Meier method was used to evaluate time to BCR and time to M for LPCa, as well as time to CR and estimation of OS for MPCa. We clearly show that PCa patients express-ing the HlA-A*02:01 allele exhibit the worst clinical outcome compared to their HlA-A*02:01 negative counterparts. Our data demonstrate, for the first time, that HlA-A*02:01 expression in the absence of HlA-A*24:02 has a significant potential as bad prognosticator for clinical outcome in PCa patients receiving the standard of care treatments according their disease status.
CONCLUSIONS: Our results reveal an unfavorable prognostic role for HlA-A*02:01 in PCa, especially in the absence HlA-A*24:02. The precise mechanisms (e.g. allelic-loss and/or immune escape or resistance) underlying this immune-based difference require further investigation.
INFLAMMATION RELATED MIRNAS AND BONE MARROW INFILTRATION IN MYELODYSPLASTIC SYNDROME (MDS) RISK DETERMINATION AND THERAPY RESPONSEC. Vaxevanis1, M. Bauer2, H.K. Al-Ali3, C. Wickenhauser2, B. Seliger11Martin Luther University Halle-Wittenberg, Institut für Medizinische Immunologie, 2Institut für Pathologie and 3Universitätsklini-kum Halle (Saale), Universitätsklinik und Poliklinik für Innere Medizin IV, Halle (Saale), Germany
Aim: Using only the currently available approaches, MDS patients risk assessment is a difficult endeavour. Next to the cytogenetic score post-transcriptional regulation might contribute to differential gene expression in this disease and has an impact on dis-ease progression and therapy response. So far, non-coding RNAs are differential expression pattern of miRNAs in AMl and MDS samples, which might have an effect on stem cell survival, maturation and immune response. Preliminary results demonstrated a differential expression of immune modulatory miRNAs in different AMl and MDS cell lines, which could be correlated to certain cytokine profiles suggesting a miRNA-dependent and -independent regulation mediated by immune escape and inflammation. Therefore, the aim of this study is to determine the role of these immune modulatory miRNAs in bone marrow (BM) biopsies of MDS patients as possible indicators of changes in the BM that might lead to AML progression.
Methods: 10 BM biopsies from healthy controls, 55 BM biopsies from MDS patients with different blast cell counts and 17 primary and sequential biopsies with secondary AML (sAML) were analysed for the expression of immune modulatory miRNAs related to inflammation (NF-kB, Il-1b, TlR4 pathways) as indicated from our MDS-AMl cell line model. Clinical parameters and cytogenetic data were available for all patients, while multiplexed immuno-histochemistry was performed for the investigation of the BM infiltrate. Clinical relevance was determined by correlating the miRNA expression pattern with BM immune cell infiltration, patient clinicopathological characteristics, treatment response and disease progression/ survival.
Results: From the various differentially expressed miRNAs within our MDS/AMl cell line model, 7 miRNAs were utilized for the creation of an expression pattern. By comparison to the median expression of the miRNAs in healthy controls, patients could be stratified into 4 distinct groups. An intermediate expression cohort (group III) included the majority of sAMl biopsies. Moreover, group Iv comprised patients with generally favorable BM infiltrates (CD8HIGHTregLOW), while group II showed a high CD4+ T cell infil-tration. Group II patients without treatment showed a tendency for a risk of reduced survival, while some group III patients failed to respond to treatments with demethylating agents leading to disease progression and/or death.
Conclusion: The combination of histopathological data with miRNA signatures might be an effective way of determining patient risk and allow for better patients’ classification. Analysis of the molecular characteristics of the BM in MDS and sAMl patients might give insights into the underlying molecular mechanisms of the altered immune cell infiltration and in the selection of a suitable therapy.
AffiliationsAdema G. Radiotherapy & OncoImmunology laboratory, Department of Radiation Oncology RadboudUMC, Ni-jmegen, The Netherlands
Bartunkova J. MD, PhD, Professor of Immunology, DrSc., Head, Institute of Immunology, 2nd Faculty of Medicine, Charles University, Prague
Baxevanis K. Scientific Director, Cancer Immunology and Immunotherapy Center “Agios Savvas”, Hospital, Athens, Greece
Brandau S. Professor, Head of the Research Division, Department of Otorhinolaryngology, West German Cancer Center, University Hospital Essen, Germany
Cavallo F. Department of Molecular Biotechnology and Health Sciences, Molecular Biotechnology Center, University of Torino, Italy
Foukas P. MD, 2nd Department of Pathology, National and Kapodistrian University of Athens, School of Medicine, ‘‘Attikon’’ University Hospital, Athens, Greece
Georgoulias V. Professor Emeritus of Medical Oncology, School of Medicine, University of Crete, GreeceGiatromanolaki A. MD, Professor of Pathology, Democritus University of Thrace, GreeceGuinn B.A. Reader in Biomedical Sciences. Department of Biomedical Sciences, University of Hull, UKInderberg E.M. Department of Cellular Therapy, Oslo University Hospital-The Norwegian Radium Hospital, Oslo, Norway
Kletsas D. Institute of Biosciences and Applications, NCSR “Demokritos”, Athens, GreeceKoinis F. Consultant of Medical Oncology, University Hospital of Larissa, Thessaly, GreeceKontopodis E. MD, Medical Oncologist, Attending Physician, Venizeleion General Hospital of Heraklion, GreeceKosmatopoulos K. Chief Executive Officer, vaxon Biotech, Paris, FranceKotsakis A. MD, PhD, Associate Professor of Medical Oncology, Director of Department of Medical, Oncology, University Hospital of Larisa, Thessaly, Greece
Koukourakis MI. MD, Professor of Radiotherapy - Oncology, Democritus University of Thrace, GreeceKoumarianou A. MD, PhD, in Tumor Immunotherapies, Imperial College, Consultant in Medical Oncology, 4th Department of Internal Medicine, Attikon University Hospital, Athens
Koutsopoulos A. MD, PhD, Associate Professor of Pathology, Laboratory of Pathology, Faculty of Medicine, University of Crete, Heraklion, Greece
Koutsoukos K. MD, MSc, Medical Oncologist, Academic Scholar, Department of Clinical Therapeutics, University of Athens, Athens, Greece
Kyrgias G. Professor of Radiation Oncology, School of Medicine, University of Thessaly, Larissa, GreeceKzhyshkowska J. PhD, Head of Department for Innate Immunity and Tolerance, Institute of Transfusion Medicine and Immunology, University of Heidelberg, Germany
Laskarakis A. Director Medical Oncologist, MD, MSc, Athens Medical Group, Athens, GreeceLucas S. Professor, Faculty of Medicine and Dentistry, de Duve Institute, UCLouvain, Brussels, BelgiumMadrigal A. Royal Free Hospital and UCL Cancer Institute, UCL Country Ambassador for MexicoMarodon G. PhD, Research Fellow, Group Leader, Center for Immunology and Infectious Diseases (CIMI-PARIS), Faculty of Medicine, Sorbonne University, Paris, France
Martinenaite E. PhD, Industrial Postdoc at IO Biotech & National Center for Cancer Immune Therapy, Denmark
Nishimura M. Research, Department of Surgery, Stritch School of Medicine Loyola University Chicago, USAPawelec G. Professor of Experimental Immunology, Center of Medical Research, Second Department of Internal Medicine, University of Tubingen, Germany
Pistamaltzian N. MD, PhD, Medical Oncologist, Consultant, Oncology University of Athens “MITERA” Hospital, Athens, Greece
Seliger B. Institute of Medical Immunology, Martin - Luther - University Halle - Wittenberg, Halle, GermanySolana R. Maimonides Biomedicine Institute of Cordoba (IMIBIC), Reina Sofia University Hospital, University of Cordoba, Spain
Stathopoulos G. Associate Professor, Department of Physiology, Principal Investigator, Laboratory for Molecular Respiratory Carcinogenesis , Faculty of Medicine, University of Patras, Greece
Stravopodis D. BSc, MSc, PhD, Associate Professor, Biology of the Cell & Development, Department of Cell Biology & Biophysics, Faculty of Biology, National and Kapodistrian University of Athens, Greece
Trougakos I. PhD, Professor, Director, Department of Cell Biology & Biophysics, Faculty of Biology, National & Kapodistrian University of Athens, Athens, Greece
Tsitsilonis O. MD, PhD, Associate Professor of Immunology, Department of Biology, National & Kapodistrian University of Athens, Greece
Tsoumakidou M. MD, PhD, Researcher B, Institute of Bioinnovation, BSRC Alexander Fleming, Athens, GreeceVan der Burg S.H. Head Experimental Cancer Immunology and Therapy, Head Research Laboratories Medical Oncology, Leiden University Medical Center, The Netherlands
Wischhusen J. PhD, Professor of Experimental Tumor Immunology, University of Würzburg School of Medicine, Department of Gynecology and Obstetrics, Germany