Day 2 :
Cairo University, Egypt
Keynote: Circulating Cell-free DNA (ccf-DNA) in cancer patients: From DNA integrity index to liquid biopsy
Time : 09:00-09:30
Azza Mahmoud Kamel is currently working as Emirate Professor of Clinical Pathology at National Cancer Institute, Cairo University, Egypt since 2007. Previously she worked as Professor of Clinical Pathology (1986- 2007) at National Cancer Institute, Cairo University, Egypt, Founder and Head of BMT Lab. Unit (1993- 2007) and Head of Clinical Pathology Department (2005-2007). She pursued her Medical Degree (MB, BCh), Faculty of Medicine, Cairo University (June 1968), Master Degree (MSc) in Clinical Pathology, Faculty of Medicine, Cairo University (October 1972) and Doctorate Degree (MD) in Clinical Pathology(Immunology/ Hematology), Faculty of Medicine, Cairo University (July 1976). She has 165 publications in many reputed journals. She conducted many workshops and completed many research projects. She received many awards like State Award in Medicine (1989), Medal of Excellence from the Egyptian Government (1994). She is an active member of Egyptian Society of Cancer, International Society of Hematology, European and African division (ISH), European Association of Hematology (EHA), American Association of Cancer Research (AACR), International Society of Thrombosis and Hemostasis (ISTH).
Cell-free indicates DNA that is found freely in the blood without a nucleus. Circulating cell-free DNA (ccf-DNAs) were first identified by Mandel and Metais in 1948 but their association with disease was not confirmed till 1977 when their increased level in the plasma/serum of cancer patients was proved. In healthy individuals, the main source of ccf-DNA is apoptotic cells which release uniform DNA fragments 185 to 200 base by a programmed enzymatic cleavage; the level is extremely variable but is usually low to negligible. Cancer cells release different and longer DNA fragments resulting from necrosis, autophagy, or mitotic catastrophe; the chance of an active release from cells was also reported and the levels are much higher than in healthy individuals. However, increased levels of ccf-DNA were observed in many diseases including leukemia, solid tumors, pulmonary embolism, myocardial infarction, tissue trauma, and chronic inflammatory diseases. This broad prevalence of diseases with potentially elevated ccf-DNA levels limits the diagnostic specificity and no cutoff value of plasma DNA concentration produced performance characteristics that would make it a good screening tool for neoplastic diseases. Thus a more refined approach was applied by calculating the ratio between cancer cell-derived ccf-DNA and normal cell-derived ccf-DNA in what is called DNA integrity index. More recently detection of tumor-specific molecular aberrations in the ccf-DNA is performed, what is called liquid biopsy. Many studies reported increased serum concentration and DNA integrity index in various solid tumors including breast, gynecological malignancy, HCC and acute myeloid leukemia. The analysis of the length of circulating DNA in plasma was reported as a sensitive marker for solid tumor detection and it was claimed to discriminate between benign and malignant lesions. The rationale of liquid biopsy is that mutations detected in ccf-DNA are highly specific of cancer and can clearly identify circulating tumor DNA (ct-DNA). Ct-DNA was explored as a prognostic or predictive marker for cancer detection; the studies suggested potential clinical applications. The analysis of ct-DNA ranges in scale from single mutations to whole-genome analyses. Liquid biopsy has many advantages compared to conventional sampling methods. The latter is subject to procedural complications, difficulty in obtaining sufficient material of adequate quality for genomic profiling and sampling biases that arise from genetic heterogeneity. A liquid biopsy is also superior to the conventional monitoring methods namely tumor markers that often lack specificity and imaging which exposes patients to ionizing radiation and has limited resolution. Promising as it is ccf-DNA and ct-DNA assays need scrupulous standardization to overlap discrepancies in sensitivities across various studies. However, sample collection is convenient, minimally invasive, and it avoids the need for tumor tissue biopsies. Analyzes of ccf- DNA and ct-DNA may have the potential to complement or replace existing cancer tissue and blood biomarkers in the future.
All India Institute of Medical Sciences, India
Keynote: Etiopathology of Wilms’ Tumor in India - Genetic Heterogeneity and Increasing “Risk Factor” involving Novel Gene Mutations of MTHFR gene polymorphism
Time : 09:30-10:00
Ajit K Saxena has received his PhD (Cytogenetic & Molecular Genetics) from Institute of Medical Sciences, Banaras Hindu University, Varanasi, (U.P) in 1989/90. After receiving his PhD degree he joined NIH funded Indo-US project at AIIMS New Delhi where he identified the role of novel antigen IL6 as a signal traducing agent in human glioblastoma. He retains more than 24 years of academic (teaching/research) experiences in various renowned Institutions of India and abroad. Currently, he is working as a Professor of Clinical Genetics and Head of the Department of Pathology & Laboratory Medicine in AIIMS Patna. Of course, he has published more than 100 articles with high citations. Based on creditability in academics’ he has received several awards and honors, including Gold Medal Award, Confer ‘Vivek Ratan Award’ and Millennium Award from USA and Excellence Award in 2014.
Wilms’ Tumor (WT) is one of the rare tumors of pediatric age group. The etiopathology of WT is complex due to interactions between genetic and environmental factors. Genetic factors include deletion of the q13 region of chromosome-11. The functional aspect of loss of genetic material is unknown but might play a significant role in the process of malignant transformation. “Risk Factors” associated with the development of WT still remain elusive. The rationale behind this study was to identify new mutations and associated “Risk Factors” in the population of Eastern part of India, an area highly susceptible for development of cancer. We performed studies at cytogenetic and molecular level to identify the genetic abnormalities associated with WT cases in comparison with the respective controls. Interestingly, a variety of different karyotypes were observed in the cases of WT that includes the loss of Y-chromosome, presence of ring chromosomes, translocation and numerical (monosomy, trisomy) variations. Further, we evaluated the frequency of WT1/WT2 gene mutation in cases and controls. PCR based analysis demonstrated that the frequency of WT1 (15%) mutation was three times higher than WT2 (5%), which may be due to interaction with environmental factors, or unknown reasons. Further, we performed Methylene Tetrahydrofolate Reductase (MTHFR) C677T gene polymorphism analysis to assess the genetic heterogeneity in WT cases using Amplification refractory mutation system (ARMS)-PCR, where allele “C” changes to “T” (C→T), increasing the “Risk” of the disease. Interestingly, DNA sequencing identified “novel mutations” in MTHFR gene, which has not been reported earlier in cases of WT. Since WT is an embryonic tumor, there may be an involvement of dysfunction of stem cells, therefore we further performed mutational studies to characterize early transcription factors (Sox-2, Oct-4, and Nanog) in WT cases. Such studies were found to be relevant to explore the transforming ability and activation of oncogenes during subtle changes observed during cytogenetics study (deletion, frameshift mutation, translocation and point mutation). The study showed differential expression of Oct-4, and mutation of Sox-2 in WT cases.
Ukrainian Anti-Cancer Institute, Austria
Time : 10:00-10:30
Wassil Nowicky, Dipl. Ing., Dr. Techn., DDDr. H. C., Director of “Nowicky Pharma” and President of the Ukrainian Anti-Cancer Institute (Vienna, Austria). Has finished his study at the Radiotechnical Faculty of the Technical University of Lviv (Ukraine) with the end of 1955 with graduation to “Diplom Ingenieur” in 1960 which title was nostrificated in Austria in 1975. He became the very first scientist in the development of the anticancer protonic therapy and is the inventor of the preparation against cancer with a selective effect on basis of celandine alkaloids “NSC-631570”. He used the factor that cancer cells are more negatively charged than normal cells and invented the Celandine alkaloid with a positive charge thanks to which it accumulates in cancer cells very fast. Author of over 300 scientific articles dedicated to cancer research. He is a real member of the New York Academy of Sciences, member of the European Union for applied immunology and of the American Association for scientific progress, honorary doctor of the Janka Kupala University in Hrodno, doctor “honoris causa” of the Open international university on complex medicine in Colombo, honorary member of the Austrian Society of a name of Albert Schweizer. He has received the award for merits of the National guild of pharmacists of America. the award of Austrian Society of sanitary, hygiene and public health services and others.
Unusual for an anticancer agent NSC 631570 possesses some distinct immune properties. The incubation of peripheral lymphocytes of healthy blood donors with NSC 631570 resulted in the increase of lymphocytes with the T-helper phenotype, decrease of the lymphocytes with T-suppressor phenotype as well as an increase of T-helper/T-suppressor ratio. NSC 631570 was administered to nine advanced stage cancer patients (4 with liver cancer, 4 with head and neck carcinomas, and one breast cancer). In three cases the tumors responded partially on the therapy, in one case a minimal response was noted, in 3 cases the disease was stable, and in 2 cases the tumors did not respond on the treatment. After the therapy, the number of T-helper cells (CD4), as well as the CD4/CD8 ratio, increased. In eight oncological patients’ immune parameters were compared before and after the treatment with NSC 631570. It was revealed NSC 631570 affected basically the thymus-dependent cells (T-cells). The number of rosette-forming T-lymphocytes was significantly higher after the treatment. No significant changes were observed in the humoral immune parameters. In nine male lung cancer, patients lymphocytes subpopulations were determined before and after the therapy with NSC 631570. The therapy resulted in increased total T-cells and a reduced T-suppressor fraction. The helper-suppressor ratio normalized. There was no sign of the activation of NK cells, T-helpers as well as B-cells. The restoration of the cellular immunity correlated with the better clinical course of the disease. The effect of NSC 631570 on the functional activity of monocytes from 20 patients with lung cancer or peritonitis was studied using nitro blue tetrazolium chloride test (NBT-test). The authors reported on the positive effect of NSC 631570 on the functional activity of the macrophages as well as antioxidant systems of monocytes and erythrocytes.23 patients with various tumors were treated with NSC 631570 and the immune e parameters were evaluated before and after the therapy. The authors observed the increase in lymphocytes and the decrease of the blood sedimentation rate. Following immune changes were also noted: increase of T-lymphocytes, T-helpers, NK- cytotoxicity, phagocytic activity, normalization of the T-helper/T-suppressor ratio, and occurrence of large granular lymphocytes.
King Chulalongkorn Memorial Hospital, Thailand
Time : 10:30-10:50
Narin Voravud has completed his MD from Meridol University and clinical residency training from the Department of Medicine, Chulalongkorn University School of Medicine, Thailand, registrar at the University of London, Hammersmith Hospital in London, United Kingdom and hemato-oncology training at the Washington University, St Louis, USA, clinical fellowship in medical oncology, MD Anderson Cancer Center, USA. He has published more than 60 papers in peer-reviewed journals and three textbooks in oncology.
Recent FDA approval for immune checkpoint inhibitors has led to the development of cutting-edge technology in an attempt to cure cancer including genetically modified adoptive immune cell therapy such as CART and TIL cell in refractory ALL and metastatic breast cancer, respectively, achieving complete remission. Unfortunately, many other tumors including metastatic pancreatic cancer is an unmet medical need without any improvement with currently available standard chemotherapy. To evaluate therapeutic efficacy and safety of adding immunotherapy to chemotherapy were retrospectively reviewed in 30 patients with metastatic adenocarcinoma of the pancreas were treated with combination systemic therapy and immunotherapy (immune checkpoint inhibitors, antiHER1 antibody, thymosin-alpha, adoptive NK/ DC vaccine, and immunonutrition) concurrently or sequentially, and compared to the control group who received standard chemotherapy regimens. Clinical benefits (RR, PFS, OS) and immunologic response (CDs, NK cell activity, IFN level) were better in the chemo-immunotarget therapy group. Concurrent treatment is better than sequential therapy. Combination treatment did not Predictive factors of tumor response include nutritional status, performance status, visceral versus lymph node metastases, PDL1 expression, TMB, MSI, and normalization of CD profiles during and after treatment. Cord blood NK cell therapy yields more durable NK cell activity and IFN level than autologous or allogeneic NK cell vaccines, whereas allogeneic NK cell therapy is better than autologous NK cell vaccine. Immunoscore, Next generation CAR-NK cell therapy and powering immune cell mitochondria are under active investigation.