Eman Tayae Elsayed is an Clinical & Research Fellow works in Department of Surgery, University College of Medicine, Egypt. Her academic interest are Hepatobiliary and pancreatic surgery / Surgical oncology for pancreatic cancer / Molecular Imaging by two-photon microscopy Pancreaticcancer,microenvironment / Cancer metabolism

Background: Peripheral blood mitochondrial DNA (mtDNA) copy number alteration has been suggested as a risk factor for several types of cancer. The aim of the present study was to assess the role of peripheral blood mtDNA copy number variation as a non-invasive biomarker in prediction and early detection of renal cell carcinoma in a cohort of Egyptian patients. Methods: Quantitative real-time polymerase chain reaction (qPCR) was used to measure peripheral blood mtDNA copy number in 57 patients with newly diagnosed, early-stage localized renal cell carcinoma and 60 age- and sex- matched healthy individuals as a control group. Results: Median mtDNA copy number was significantly higher in renal cell carcinoma cases than in controls (166 versus 91, P<0.001). Increased mtDNA copy number was associated with 18-fold increased risk of RCC (95% confidence interval: 5.065-63.9). On using receiver operating characteristic curve analysis, it was found that mtDNA could distinguish between RCC patients and healthy controls with 86% sensitivity, 80% specificity, 80.3 % positive predictive value and 85.7% negative predictive value at cutoff value of 108.5. Conclusion: This study suggests that increased peripheral blood mtDNA copy number could be used as a potential independent predictor of RCC risk. In addition, it may serve as a promising non-invasive biomarker for early detection of RCC.

Mai Moaaz is an assistant professor of Immunology. She has a passion in the field of tumor immunology and an expertise in tumor tissue culture system and experimenting new modalities for cancer immunotherapy. Her tissue culture system provides the ex vivo tissue architecture that is necessary to maintain or reconstitute an environment closely resembling that of the tumor tissue to reflect the complex tissue architecture of an individual tumor. The majority of preclinical cancer research is based on established cell lines that frequently have undergone multiple changes influencing their biological behavior and therefore no longer reflect that of the primary tumor of origin. She has built this model after years of experience in research, evaluation, teaching and administration both in hospital and education institutions. She is conducting current studies on tumors as an approach to cancer immunotherapy.

Statement of the Problem: Because of heterogeneity of gastric cancer (GC), searching for more accurate predictors of GC prognosis has become a growing interest. Infiltration of immune cells in tumors is associated with prognosis; among them are the immunosuppressive myeloid derived cells (MDSCs). Their accumulation constitutes an important mechanism of tumor immune evasion. Hence, their characterization is essential for diagnosis and therapeutics of cancer. Increased expression of microRNA-494 was noticed in MDSCs from tumor-bearing mice suggesting another new therapeutic objective for cancer treatment. It was also discovered that tumor-derived transforming growth factor beta (TGF-β) was responsible for the up-regulation of microRNA-494 in MDSCs. The purpose of this study is to address the suppressive effect of MDSCs on T cells in GC and its possible association with Micro-RNA-494 and TGF-β expression in tumor tissue. Methodology & Theoretical Orientation: A case control study on 40 GC patients and corresponding controls where done. Tissue samples and peripheral blood were used for isolation of CD33+11b+HLADR- MDSCs cells using flowcytometry. MDSCs were co-cultured with isolated T cells to assess proliferation and cytokine production. Real-time PCR and Enzyme linked immunosorbent assay were used to evaluate tumor expression of miRNA-494 and TGF-β respectively. Findings: MDSCs percentages were significantly elevated in GC patients than controls and significantly increased in tumor specimens than paraneoplastic tissue. This increased expression is accompanied with elevation of TGF-β production in tumor than surroundings. MiRNA-494 was also extensively expressed in tumor samples. Addition of MDSCs from cancer patients markedly suppressed the proliferation of autologous T cells and cytokine production; however, no inhibitory effect was observed for MDSCs from healthy donors. Conclusion & Significance: The result indicates that tumor-derived MDSCs but not MDSCs from healthy donors have the immunosuppressive effect on T cells. Infiltration of MDSCs in tumors is associated with the prognosis of GC.

Mai Moaaz is an assistant professor of Immunology. She has a passion in the field of tumor immunology and an expertise in tumor tissue culture system and experimenting new modalities for cancer immunotherapy. Her tissue culture system provides the ex vivo tissue architecture that is necessary to maintain or reconstitute an environment closely resembling that of the tumor tissue to reflect the complex tissue architecture of an individual tumor. The majority of preclinical cancer research is based on established cell lines that frequently have undergone multiple changes influencing their biological behavior and therefore no longer reflect that of the primary tumor of origin. She has built this model after years of experience in research, evaluation, teaching and administration both in hospital and education institutions. She is conducting current studies on tumors as an approach to cancer immunotherapy.

Statement of the Problem: Breast cancer is the most common cancer in females being the leading cause of cancer mortality in women. The continuous interactions of cancerous cells with their surrounding microenvironment infiltrating immune cells shape their immunogenicity as well as the ability to tumor progression and metastasis. Interleukin 17A (IL-17A) has a promoting role in carcinogenesis, tumor metastasis and resistance to chemotherapy. It enhances tumor cell survival and invasiveness and inhibits the antitumor immune response. Pro-tumor effects of IL-17A may occur via an increase in suppressive functions of myeloid-derived suppressor cells (MDSCs). The expression of IL-17A and programmed death ligand 1 (PDL1) is increased in breast cancer. The PDL1–PD1 (programmed death protein 1) signaling pathway promotes escape from immune surveillance in tumor cells. The purpose of this study is to address the potential suppressive effects of monoclonal anti-IL-17 antibodies on IL-17 tumorigenic activities in intact tumor microenvironment of BC as a novel immunotherapeutic reagent. Methodology & Theoretical Orientation: Fresh tumor tissue samples and peripheral blood were taken from 50 BC patients. IL-17A and PDL1 expression were primarily assessed. Cultures of tumor tissues either supplemented or not with anti-IL-17 monoclonal antibodies were subjected to evaluation of PDL1, MDSCs and T cell activities. Findings: Our results revealed that IL-17A stimulated PDL1 expression in tumor BC cultures and were correlated to clinicopathological features. Culturing with anti-IL-17 monoclonal antibodies suppressed PDL1 expression, and promoted T cell proliferation and functions. Tumors cultured with anti-IL-17 showed a significant decrease in the expression of MDSCs cells in the tumor microenvironment. Conclusion & Significance: The results indicate that recombinant anti-IL-17 monoclonal antibodies could represent a novel effective element of immunotherapeutic treatment strategy for BC. The selectivity and anti- potential of anti-IL-17 is highly hopeful in IL-17- abundant BC tumor microenvironment.

Naoki Otsu is interested in stem cell biology and tumorigenesis from stem cells. Based on Cancer stem cell theory, we are searching for cells with high tumorigenicity and membrane proteins intensely expressed in the cells. Currently, we focus on glioblastoma, a malignant brain tumor, and analyze membrane proteins that are molecular targets of antibody drugs. Using induced cancer stem cells derived from neural stem cells as model cells, strongly expressed membrane proteins were searched for multiple candidate proteins. This presentation introduces the molecules whose function and molecular mechanism are clarified among the obtained candidate.

Research focused on the property and targets of cancer stem cells, which have tumorigenicity and treatment-resistant, may contribute to the discovery of new therapeutic and diagnostic strategies. We have previously established mouse glioma-initiating cell (mGIC) lines from normal neural lineage cells, analyzed their gene expression profile, and reported functions of GIC-specific genes. We further focused on GIC-specific membrane proteins and found uncharacterized cell-membrane protein Eva1. Here we show the characterization of Eva1 in GIC: Eva1 is highly expressed in mouse GIC line, NSCL61, and human glioblastoma (GBM, WHO grade IV)-derived floating spheres, whereas it is not expressed in normal adult mouse and human brain. Eva1 expression is correlated with the malignancy of glioma. Indeed, we found that Eva1+ cells disseminate in human GBM tissue. Moreover, knockdown of Eva1 inhibited cell proliferation and tumorigenicity in mGIC and hGIC. Forced-expression of Eva1 enhanced malignancy of Low-grade human glioma (Diffuse Astrocytoma: DA, WHO grade II). The combination of polyclonal anti-Eva1 antibody and cytotoxic factor Saporin kills GIC, therefore it promised as a new target of antibody preparation. Using gene expression profiles between mGIC and Eva1-knockdown mGIC, we further found Eva1 signal pathway. Eva1 induced GIC proliferation through the activation of the RelB-dependent noncanonical NF-kB pathway by recruiting TRAF2 to the cytoplasmic tail. This signal pathway is important for human GBM formation. Taken together, these findings suggest that Eva1 is a potential target for immunotherapy.

Chunman Lee has his expertise in evaluation and passion in improving the antitumor efficacy of novel formulations against cancer dissemination. He started carrier as a surgeon at Osaka university hospital, and passed through the surgery program of Osaka university graduate school of medicine. Then he worked at University of Texas Southwestern Med. Ctr. at Dallas as a postdoctoral fellow. Now he researches the immunotherapy against pleural mesothelioma which is very refractory disease at Osaka university hospital as an associate professor.

Hemagglutinating virus of Japan envelope (HVJ-E) developed from HVJ virus. HVJ was first discovered in Japan in 1950’s. It is mouse parainfluenza virus, not human pathogen. The virus has become very famous, since the discovery of virus-mediated cell fusion. Using the fusion activity of HVJ, a versatile drug delivery system mainly gene transfection was developed. Recently we discovered that HVJ-E possesses the various antitumor activities. One is enhancing antitumor immunities such as activation of dendritic cells, induction of natural killer cells and cytotoxicic T lymphocytes, and suppression of regulatory T cells. Other activities are direct tumor-killing by the induction of cell death through the RIG-I/MAVS pathway. We already finished phase I trial against melanoma and castration resistant prostate cancer. Then, we did the phase I dose escalation safety/tolerability and preliminary efficacy study of intra-tumoral and subsequent subcutaneous administration of HVJ-E to the patients suffering from chemotherapy–resistant pleural mesothelioma who had not receive chemotherapy in the last 4 weeks, had a performance status of 0-1, had certain functions of bone marrow, liver, kidney, and lung, had evaluable lesion with Computed Tomography and FDG-PET scan, had lesion which can be administrated with HVJ-E. Exclusion criteria is presence of autoimmune disease, interstitial pneumonia needed to treat, and other malignant lesions, use of systemic steroids, a protocol is consisted of initial-tumor administration of HVJ-E and the subsequent three subcutaneous administrations within two weeks, and then washed out for two weeks. This one cycle is repeated twice. Six patients were registered, and they were finished receiving this clinical trial. The adverse events were evaluated by independent data monitoring committee. We will show the data of AEs and antitumor efficacy of HVJ-E in this clinical trials.

Nickolai Usachev has substantial practical expertise in cardiology and intensive care, combined with more than decade’s experience in the field of clinical studies. Since 2009 Dr.Usachev heads the Pharmacovigilance unit of PSI CRO.

Summarizing our experience of medical monitoring in clinical studies in patients with relapsed diffuse large B-cell lymphoma (DLBCL) pre-treated with standard Rituximab based scheme, we were surprised by a rather high incidence of increased Troponin T (TnT) in patients that had no detectable cardiac pathology. Such patients were screened out even if this lab finding was the only contraindication for inclusion. Purpose of the study: accumulate literature data confirming that in this particular group of patients mildly elevated TnT measured by immunoassay and not supported by any overt cardiovascular manifestations can be caused by reasons other than cardiovascular pathology. Therefore such patient can be included in the study and have potential benefit from treatment. Findings: about 10-11% of patient’s serum samples [1] may contain HAMA (human anti-mouse antibodies) [2], which lead to false positive results of troponin immunoassay [3; 4; 5]. Oncology patients, especially treated with monoclonal antibodies (Mab)–based immunotherapy are at significantly higher risk of HAMA development [2; 6; 7]. Available literature data suggest that quantitative measurement of HAMA in oncology patients has far more practical impact than simple confirmation of ambiguous immunoassay results. Survival benefit associated with HAMA in patients with B-cell malignancies [8] and with non-Hodgkin’s lymphoma treated with anti-lymphoma Mab [9; 10; 11] had been demonstrated. Conclusion: HAMA measurement in potentially eligible patients who had previously received Mab immunotherapy and whose only contraindication for inclusion into the clinical study is mildly elevated TnT, seems justified. First, the patients with the false-positive TnT results caused by HAMA, will be included and may have potential benefits from study treatment. Second, the result of the HAMA test may have predictive value for treatment benefit.

Guang-Yaw LIU, PhD., Professor, Institute of Biochemistry, Microbiology & Immunology, Chung Shan Medical University.

Peptidylarginine deiminase type 2 (PADI2) is a post-translational modification enzyme that catalyzes arginine residues into the citrulline residues. Previous studies have shown that PADI2 promotes protein citrullinations in lymphocytes and it might play an important role in cell-mediated inflammation. We have found that overexpression of PADI2 promotes apoptosis in activated T cells previously. Whether does PADI2 participate in the pathway of activated T cell autonomous death (ACAD) is still curious. In this delicate PADI2-mediated ACAD study, we found that overexpression of PADI2 displayed higher levels of citrullinated protein which would induce the ER stresses significantly. The high levels of citrullinated protein results in unfolding protein response (UPR) of ER stresses and increases the huge protein degradation loading. Autophagy might embrace the engulfment and degradation capacity of the citrullinated and unfolding proteins. Herein, PADI2 could enhance autophagy in Jurkat T cells and lead to a degradation of p62 and the accumulation of LC3-II, BCEN1, ATG5 and ATG12. Autophagy and apoptosis are two critical mechanisms both which participate against cellular stresses and decide T cell activation, survival and immuno-homeostasis. PADI2-overexpressed Jurkat T cells caused the activation of Th17 cells due to the increase mRNA expression of cytokines, such as IL-17, IL-21, IL-22 and TNFα. Cytokines declined autophagy, provoked caspase cascade expression, and led to ACAD by IL-6 shRNA inspection. Simultaneously, autophagic BCEN1 could reduce Bcl-xL expression, increase caspase cascade and cause to cell insults. Knockdown of BCEN1 possibly will rescue Jurkat T cell activation, increase cytokine release and induce ACAD. We suggested that PADI2 participated in the activated T cell-induced autonomous death through triggering ER stress pathway coupling with regulating autophagic processing, and stimulating Th17 activation and the expression of cytokines by PADI2-citrullinating mechanism.