Day 2 :
Keynote Forum
Jianhua Luo
University of Pittsburgh School of Medicine, USA
Keynote: Cancer genomics and its role in human cancers
Time : 10:00-10:30
Biography:
Jianhua Luo has been studying molecular mechanism related to human malignancies in the last 24 years. Currently, he is a Professor of Pathology and Director of High Throughput Genome Center at University of Pittsburgh. In the last 16 years, he has been largely focusing on genetic and molecular mechanism of human prostate cancer and hepatocellular carcinomas. He has characterized several signaling pathways that play critical role in prostate cancer development. He proposed prostate cancer field effect in 2002 and he is one of the pioneers in utilizing high throughput gene expression and genome analyses to analyze fi eld effects in prostate cancer and liver cancer. He is also the fi rst in using methylation array and whole genome methylation sequencing to analyze prostate cancer. Recently, his group found that patterns of copy number variants of certain specific genome loci are predictive of prostate cancer clinical outcomes, regardless tissue origin. His discovery of several novel fusion transcripts and their association with aggressive prostate cancer has brought signifi cant new insight into the fi eld of prostate cancer research. Overall, these findings advance our understanding on how cancer develops and behaves and lay down the foundation for better future diagnosis and treatment of human malignancies
Abstract:
Cancer remains one of the most lethal diseases for human. In recent studies, we performed whole genome analysis on prostate and liver cancers. Our result showed that combination of genome copy number variance, genome methylation pattern and novel fusion transcripts specifi c for cancer achieved high accuracy in predicting clinical outcomes of these cancers. Interestingly, some of these fusion genes are also present in a variety of human malignancies. Some of these fusion gene products trigger new pathways that are essential for carcinogenesis in multiple human cancers, and create novel functions that are not present in wild type gene counterparts. Some of these novel fusion genes are highly targetable and treatment of cancers with drugs specifi c for these genes and their signaling pathways produced dramatic improvement of metastasis and survival rate of animals xenograft ed with cancers positive for these fusion genes.Our analyses suggest that targeting therapy for fusion genes holds promise as an eff ective treatment for human cancers.
Keynote Forum
Isabel Desgagné-Penix,
Université du Québec à Trois-Rivières, Canada
Keynote: Systems biology and metabolic engineering of microalgae for the production of pharmaceutical Amaryllidaceae alkaloids
Time : 10:30-11:00
Biography:
Isabel Desgagné-Penix has her expertise in Plant Biochemistry and specialized metabolism. Her research program is aimed at understanding the Molecular Biology and Biochemistry of isoquinoline alkaloid metabolism in Amaryllidaceae plants. Narcisses and snowdrops remains the only commercial source for the anti-cholinesterase galantamine and several potential alkaloid pharmaceuticals. The recent availability of some Amaryllidaceae alkaloid biosynthetic genes creates metabolic engineering opportunities in plants and microorganisms. Her innovative and creative new ways to attempt production of valuable plant metabolites are of interest.
Abstract:
Several Amarylllidaceae plant alkaloids (AAs) possess powerful pharmaceutical and biotechnological properties. Thus, AA metabolism and its fascinating molecules, including anti acetylcholinesterase galantamine, anti-microbial lycorine and anti-cancer narciclasine, have attracted the attention of both the industry and researchers involved in plant science, chemical bioengineering and medicine. Currently, access and availability of high-value AAs [commercialized (e.g. galantamine) or not (e.g. narciclasine)] is limited by low concentration in nature, seasonal production and time-consuming low-yield extraction methods. Nevertheless, commercial AA galantamine is still extracted from plant sources. Eff orts to improve the production of AA have largely been impaired by the lack of knowledge on AA metabolism. Th e purpose of this study was to use recent development and integration of next-generation sequencing technologies and metabolomics analyses to unravel metabolic pathways allowing the use of metabolic engineering and synthetic biology approaches to increase production of valuable AAs. Novel genes encoding AA biosynthetic enzymes were identifi ed from our transcriptome databases using bioinformatics tools. The genes were characterized, and their activities were studied through classical biochemistry experiment such as cloning into expression vectors, heterologous expression, recombinant protein purifi cation and specifi c enzyme assays. In addition, AA precursor pathway was introduced into microalgae cells to validate the function of the biosynthetic genes and to produce AA metabolites (precursors and intermediates). Next, the final steps of the AA biosynthetic pathway will be added to reach galantamine or other AA synthesis in microalgae. Metabolic engineering provides opportunity to overcome issues related to restricted availability, diversifi cation and productivity of plant alkaloids. Engineered cells can act as iofactories by off ering their metabolic machinery for the purpose of optimizing the conditions and increasing the productivity of a specific alkaloid.
- Cancer Biomarkers | Biomarkers & Immuno-Oncology | Biomarkers Detection & Discovery | Clinical Biomarkers | Advances in Biomarkers Discovery
Location: Burgos
Chair
Topolcan Ondrej
Charles University, Czech Republic
Session Introduction
P G Lokhov
Institute of Biomedical Chemistry, Russia
Title: Usage of metabolomics profile as biomarkers itself for diagnostic diseases
Biography:
P G Lokhov is the Head of Laboratory for Mass Spectrometry-based Medical Metabolomics in the Department of Proteomic Research and Mass Spectrometry of IBMC, a position he has held since 2012. He is mainly interested in mass spectrometry-based metabolomics and proteomics, and their application for diagnostics. He graduated from the 2nd Moscow Medical Institute, took the post-graduate course and thereafter received a PhD degree in Biochemistry in 2002. During his career Dr. Lokhov has received several awards, including the State Prize of the Government of Russia.
Abstract:
Recently, the researches to fi nd proteins as biomarkers of various diseases have become widespread. Especially these studies are popular in оncology. However, until now proteins specifi c for cancer cells are not detected. In spite of it, to diagnose cancer diseases biomarkers which are specifi c for a neoplastic process generally are widely used (such as CA-125, CA 19-9, CA 15-3, PSA, α-fetoprotein etc.). However, they can’t discharge the main task: to diagnose the initial stages of disease, because their opportunity is limited by low sensitivity of existing laboratory technologies. Th e situation is much better in the case of using RNA’s as biomarkers, because PCR takes off such restrictions. In the case of metabolomic diagnostics there are the same concentration limits as in proteomics. Th erefore, it was decided not to use separate low molecular weight compounds as biomarkers but the whole metabolomic profile of the sick person as a biomarker itself of a disease. With this purpose, at the fi rst stage that profi le was clusterized using the method of principle component analysis to detect clusters of metabolites which are diff erent from the clusters of healthy person. At the second stage, the specifi city and sensitivity of the proposed technology was evaluated using the SVM and ROC analyses, for diagnostics of lung, prostate and other cancers.
Biography:
Mercedes Piedad de León Bautista has completed her Bachelor’s Degree from UPAEP University School of Medicine, and an MSc and PhD from CINVESTAV-IPN in the Dept. of Molecular Biomedicine. She is the Medical Director and Laboratory Chief of Central ADN, a molecular laboratory focusing on human health and translational medicine. She has been working with PDAC and, nowadays, her biomedical efforts are based on molecular platforms to fi nd new markers in cancer and hereditary diseases.
Abstract:
Pancreatic ductal adenocarcinoma (PDAC) is a complex pathology with poor prognosis. Eff orts have been focused on understanding the role of RhoGDI's in PDAC, in particular, RhoGDI1 and RhoGDI2. However, the role of RhoGDI3 has neither been studied in relation to cancer nor to PDAC. Our group have characterized the expression and functionality of RhoGDI3 and its target GTPases, RhoG and RhoB, in pancreatic cell lines and compared it to human tissue. Th rough immunofl uorescences, pull down assays, subcellular fractionation and immunohistochemistry, we found a reduction in RhoGDI3 expression in PDAC late stages, and this reduction correlates with tumor progression and aggressiveness. Despite the reduction in the expression of RhoGDI3 in PDAC, we found that RhoB was under expressed while RhoG was over expressed, suggesting that cancerous cells keep their capacity to
activate this pathway, thus these cells may be eager to the stimuli needed to proliferate and become invasive. Surprisingly, we found nuclear localization of RhoGDI3 in non-cancerous pancreatic cell line and normal pancreatic tissue biopsies, which could open the possibility of novel nuclear functions for this protein, impacting gene expression regulation and cellular homeostasis. To elucidate the possible functions of RhoGDI3 in cancer maintenance, the overexpression assays have demonstrated that increased RhoGDI3 protein increases proliferation rate; besides, the xenograft tumor was smaller compared to the mock, suggesting and predicting that overexpression of RhoGDI3 is an important molecule to constrain the tumoral volume. In conclusion, RhoGDI3 protein decreases the malignant behavior in PDAC.
Topolcan Ondrej
Charles University, Czech Republic
Title: PHI and prostate cancer - optimal management
Biography:
Topolcan Ondrej has graduated from the Medical Faculty, Charles University in Pilsen. He is working in the Dep. of Nuclear Medicine Faculty Hospital in Pilsen
and he was the Head of Imunoanalytic Laboratory in 2000. He is a Member of Scientific Committee Endocrinology Institute Prague and he was awarded the Carl
R Jolliff award for lifetime achievement in Clinical and Diagnostic Immunology by the American Association for Clinical Chemistry (AACC) in 2011.
Abstract:
Background: Prostate cancer is the fi ft h leading cause of cancer death among men. More than one million cases are worldwide diagnosed every year, and the mortality is over 300,000 deaths per year. Prostatic specifi c antigen (PSA) is a serine protease composed of 240 amino acids in a single polypeptide chain. In serum, PSA is primarily bound to alpha1-anti-chymotrypsin (95% of total PSA), to a lesser extent to alpha2-macroglobulin. 10-30% serum PSA occurs in the free form (fPSA). Since the 80s of the last century, total level of prostatic specifi c antigen (tPSA) is used as a tumor marker in the prostate cancer diagnostics. Most of the prostate tumors are diagnosed using biopsy based on the elevated levels of PSA. It is also useful to determine the part of PSA called proPSA. This part of PSA is more produced by tumor cells. Based on the levels of PSA, proPSA, and free PSA, the prostate health index PHI can be calculated.
Aim: Demonstration of the usefulness of PHI in the management of prostate cancer patients.
Materials & Methods: Cohort of 1865 patients was evaluated. Th e total of 5800 biopsies, 1448 MRI and 150 PET/MRI were performed in this cohort of patients. 4900 samples with the results of PSA, fPSA, -2proPSA, fPSA/tPSA calculated results and PHI during the follow-up period. Gleason score was established in all patients.
Results: By comparison of AUC sensitivity for each laboratory parameter, PHI achieved the highest value (0.8118). PHI achieved also the best correlation with the Gleason score (G6-G9) which allows PHI to be a reliable marker of aggressiveness of the prostate cancer. An optimal PHI cut-off value was 31 with specifi city 20.4%, sensitivity 97.6%, PV+ 18%, PV- 97%, RR 8.7.
Conclusions: Determining PHI allows proposing the optimal diagnostic algorithm for prostate cancer, improvement of diff erential diagnosis of carcinoma vs. prostate hypertrophy, reducing biopsies and imaging techniques, more accurate prognosis estimation, optimizing the type of surgery, optimizing of the postoperative treatment and optimizing of follow-up.
Yeng S Ang
Salford Royal University Hospital, UK
Title: PEA3/ETV4-related transcription factors coupled with active ERK signalling are associated with poor prognosis in gastric adenocarcinoma
Biography:
Yeng S Ang has an international professional standing and research expertise to enhance clinical interventions in Barrett's oesophagus and oesophageal cancer. He is a Member of the BSG/National Clinical Research Institute Upper GI early cancer prevention research subgroup. He is a peer Reviewer for the NIHR RFPB programme and a member of the Research Steering Board of Manchester Cancer Research Centre (Cancer Research UK Manchester Institute). These research initiatives have shaped his contribution for the management of GORD, Barrett’s oesophagus and oesophageal cancer. He has published over 45 articles and he is a Supervisor for PhD and MD students in the molecular cancer group of the University of Manchester.
Abstract:
Background: Transcription factors often play important roles in tumorigenesis. Members of the PEA3 subfamily of ETS-domain transcription factors fulfill such a role and have been associated with tumor metastasis in several diff erent cancers. Moreover, the activity of the PEA3 subfamily transcription factors is potentiated by Ras-ERK pathway signalling, which is itself often deregulated in tumor cells.
Methods: Immunohistochemical patterns of PEA3 expression and active ERK signalling were analyzed and mRNA expression levels
of PEA3, ER81, MMP-1 and MMP-7 were determined in gastric adenocarcinoma samples.
Results: Here, we have studied the expression of the PEA3 subfamily members PEA3/ETV4 and ER81/ETV1 in gastricadenocarcinomas. PEA3 is upregulated at the protein level in gastric adenocarcinomas and both PEA3/ETV4 and ER81/ETV1 are upregulated at the mRNA level in gastric adenocarcinoma tissues. This increased expression correlates with the expression of a target gene associated with metastasis, MMP-1. Enhanced ERK signalling is also more prevalent in late-stage gastric adenocarcinomas and the co-association of ERK signalling and PEA3 expression also occurs in late-stage gastric adenocarcinomas. Furthermore, the coassociation of ERK signalling and PEA3 expression correlates with decreased survival rates.
Conclusions: This study shows that members of the PEA3 subfamily of transcription factors are upregulated in gastric adenocarcinomas and that the simultaneous upregulation of PEA3 expression and ERK pathway signalling is indicative of late-stage disease and a poor survival prognosis.
Varda Shoshan-Barmatz
Ben-Gurion University of the Negev, Israel
Title: Novel biomarker proteins for cancer: Impact on diagnosis, prognosis and treatment
Biography:
Varda Shoshan Barmatz is the Hyman-Kreitman Chair in Bioenergetics at Ben-Gurion University (BGU). Her PhD is from the Weizmann Institute; Post-doctoral training was from the University of Wisconsin-Madison and the University of Toronto. In 1982, she joined the Department of Life Sciences, BGU, and served as Chair (2000-2004). Her awards include the Hestrin Prize of the Israel Biochemistry Society, Teva Award for Young Scientists (1993) and Teva Founders Award (2016). Lady Globes magazine selected her as one of the 50 most infl uential women in Israel (2009) and one of five who made breakthroughs in science (2016). She was the Founder and Director (2006-2015) of the National Institute for Biotechnology in the Negev.
Abstract:
Cancer cells undergo re-programing of metabolism, cell survival and anti-apoptotic defenses, with the proteins mediating these re-programing representing potential biomarkers. Here, using specific antibodies, mass spectrometry and bioinformatics tools; we searched for novel biomarker proteins in chronic lymphocytic leukemia (CLL) and in non-small cell lung cancer (NSCLC) patient samples that can impact diagnosis, treatment and prognosis. By comparing protein expression profi les of CLL- and healthy donorderived lymphocytes, we identified 1,360 diff erentially expressed proteins, some shown for the first time to be associated CLL. Downregulated expression of two proteins resulted in cell growth inhibition, pointing to their essential functions. Based on changes in the levels of several proteins in CLL patients, we could distinguish between patients in a stable disease state and those who would be later subjected to anti-cancer treatments, 2-3 years before the physician’s decision. In NSCLC, the adenocarcinoma (AC) and squamous cell carcinoma (SCC), sub-types present unique genomes, transcriptomes, and proteomes, and share clinical and histopathological characteristics, yet diff er in treatment. We identifi ed novel biomarker proteins in NSCLC, with 378 proteins showing a ≥|100|-fold change in level. Several, identifi ed for the fi rst time, allow for distinguishing between AC and SCC. These, together with markers previously proposed and confi rmed here, lead us to propose a list of proteins for discriminating SCC and AC, with four being secreted. Precise diagnosis of AC and SCC is essential for selecting appropriate treatment. Finally, some of these biomarkers can serve as new targets and lead to new treatments for lung and CLL cancers.
M Al Hassan
Beirut Arab University, Lebanon
Title: Soarfenib effect on human colon cancer cells HCT116 and HCT116 p53-/-
Biography:
M Al Hassan is a current PhD candidate at the Beirut Arab University, working in collaboration with the American University of Beirut on Sorafenib and its in-vitro
and in-vivo effects on colorectal cancer. She has graduated from the Lebanese American University with a Master’s degree in Cells and Molecular Biology. Her MS thesis was about the effect of metformin on the metastasis and the 3D motility of glioblastoma cancer cells.
Abstract:
Sorafenib, a kinase inhibitor, has been approved among the drugs for the treatment of radioactive iodine resistant thyroid carcinoma, primary kidney and liver cancer. Reported targets of sorafenib include VEGFR, af family, and PDGFR belonging to the general class of tyrosine kinases. Blocking growth signals in kidney and breast cancers underlie one of the mechanisms of sorafenib antitumor effects’ leading to cell death. We hereby examine the effect of sorafenib on human colon carcinoma cell-line HCT116. We also investigate the possible role of p53 in mediating this eff ect using mutant HCT116 p53-/- cells. Cultured wild and mutant cells are treated with sorafenib (0-75μM) for 24 hr. Th is is followed by assessing the viability of cells using MTT and trypan blue exclusion assays. We also examined if sorafenib mode of action is mediated by ROS. Levels of ROS were determined in the presence and absence of antioxidants using the colorimetric NBT assay. Our preliminary results show a concentration dependent decrease in viability (trypan blue) with an estimated EC50 of 10 and 25 μM for HCT116 and HCT116 p53-/- respectively. Compared to trypan blue, MTT results were similar in case of HCT116 p53-/- but were signifi cantly diff erent with HCT116. Furthermore we obtained a significant increase in level of ROS of: 37.11% and 31.30% for HCT116 and HCT116p53-/- respectively. However, 2 hr. pre-incubation of cells with antioxidants, Trolox, N-acetylcysteine (NAC), and catalase, prior to sorafenib treatment, exerted no diff erent eff ect. No restoration of viability or decrease in generated ROS level was noted. Our preliminary fi ndings show that sorafenib action is independent of ROS level and p53 expression and further investigations on the mechanism(s) of sorafenib action are ongoing.
Sayda Omer Ebnaof
University of Khartoum, Sudan
Title: Diagnostic and prognostic microRNAs in the serum of breast cancer patients measured by droplet digital PCR
Biography:
Sayda Omer Ebnaof has completed her PhD in Molecular Biology from University of Ferrara, Italy in 2016 and MSc in Molecular Medicine from Institute of Endemic Diseases, University of Khartoum-Sudan. She is a Lecturer of Histopathology and Cytology at University of Khartoum. She has published four papers in reputed
journals.
Abstract:
Background: Breast cancer circulating biomarkers include carcinoembryonic antigen and carbohydrate antigen 15–3, which are used for patient follow-up.Since sensitivity and specifi city are low, novel and more useful biomarkers are needed. The presence of stable circulating microRNAs (miRNAs) in serum or plasma suggested a promising role for these tiny RNAs as cancer biomarkers. To acquire an absolute concentration of circulating miRNAs and reduce the impact of preanalytical and analytical variables, we used the droplet digital PCR (ddPCR) technique.
Results: We investigated a panel of fi ve miRNAs in the sera of two independent cohorts of breast cancer patients and disease-free
controls. The study showed that miR-148b-3p and miR-652-3p levels were significantly lower in the serum of breast cancer patients than that in controls in both cohorts. For these two miRNAs, the stratifi cation of breast cancer patients versus controls was confirmed by receiver operating characteristic curve analyses. In addition, we showed that higher levels of serum miR-10b-5p were associated with clinicobiological markers of poor prognosis.
Conclusions: Th e study revealed the usefulness of the ddPCR approach for the quantification of circulating miRNAas.The use of the ddPCR quantitative approach revealed very good agreement between two independent cohorts in terms of comparable absolute miRNA concentrations and consistent trends of dysregulation in breast cancer patients versus controls. Overall, this study supports the use of the quantitative ddPCR approach for monitoring the absolute levels of diagnostic and prognostic tumor-specific circulating miRNAs.
Maimoona Sabir
University of Haripur, Pakistan
Title: Association of human papilloma virus with head and neck cancer patients from Khyber Pakhtunkhwa, Pakistan
Biography:
Maimoona Sabir has completed her PhD from the Department of Biosciences, COMSATS, Institute of Information Technolgy, Islamabad, Pakistan. She is currently working as Assistant Professor in the Department of Microbiology, University of Haripur, KP, Pakistan. She has published more than 15 reserach papers in reputed journals. She has won grant from Higher Education Commssion of Pakistan for research work, “role of HPV in head and neck cancer in Pakistani population”.
Abstract:
Head and neck cancer (HNC) is the sixth most common cancer causing a high mortality rate worldwide. There are number of risk factors involved in head and neck cancer such as tobacco, alcohol, radiations, inhaling asbestos, wood dust, paint fumes and viruses. Most common virus involved in the subset of head and neck cancer is human Papilloma virus (HPV). Human papilloma virus is double stranded DNA virus which is about 8Kb surrounded by protein coat consisting of two proteins L1 and L2. Out of 100 diff erent types of HPV 15 are considered as highly pathogenic. HPV is divided into two major types; High risk HPV and Low risk HPV. In current study 150 HNC patients were screened for HPV genotyping and risk factors possibly associated with HNC. DNA was extrracted by standard phenol chloroform extraction method followed by HPV genotyping by polymerase chain reaction and agarose gel electrophoresis. In study, among the HNC aff ected individuals 81(54%) were HPV+ive and 69 (46%) HPV–ive. Statistically, no significant diff erence was observed between HPV+ive and HPV-ive. HNC cases in terms of gender. In terms of age group, statistically no significant diff erence was observed between two groups (age group >40, <40). With respect to anatomic site of head and neck cancer patients, highest number of HPV+iv was observed in oral cavity 51(63%) followed by pharynx 20(25%), larynx 7(8%) and hypopharynx 3(4%). Disease was more common among illiterate individuals with low socioeconomic status and trend of HPV+ive HNC cases have increaed from year 2011 to 2016. It can be concluded from the data that HPV infection can be the main risk factor for HNC cases in Pakistan. Further research is needed to elucidate the mechanism involved in HPV infection with head and neck carcinogenesis.
Maimoona Sabir
University of Haripur, Pakistan
Title: Association of human papilloma virus with head and neck cancer patients from Khyber Pakhtunkhwa, Pakistan
Biography:
Maimoona Sabir has completed her PhD from the Department of Biosciences, COMSATS, Institute of Information Technolgy, Islamabad, Pakistan. She is currently working as Assistant Professor in the Department of Microbiology, University of Haripur, KP, Pakistan. She has published more than 15 reserach papers in reputed journals. She has won grant from Higher Education Commssion of Pakistan for research work, “role of HPV in head and neck cancer in Pakistani population”.
Abstract:
Head and neck cancer (HNC) is the sixth most common cancer causing a high mortality rate worldwide. There are number of risk factors involved in head and neck cancer such as tobacco, alcohol, radiations, inhaling asbestos, wood dust, paint fumes and viruses. Most common virus involved in the subset of head and neck cancer is human Papilloma virus (HPV). Human papilloma virus is double stranded DNA virus which is about 8Kb surrounded by protein coat consisting of two proteins L1 and L2. Out of 100 diff erent types of HPV 15 are considered as highly pathogenic. HPV is divided into two major types; High risk HPV and Low risk HPV. In current study 150 HNC patients were screened for HPV genotyping and risk factors possibly associated with HNC. DNA was extrracted by standard phenol chloroform extraction method followed by HPV genotyping by polymerase chain reaction and agarose gel electrophoresis. In study, among the HNC aff ected individuals 81(54%) were HPV+ive and 69 (46%) HPV–ive. Statistically, no significant diff erence was observed between HPV+ive and HPV-ive. HNC cases in terms of gender. In terms of age group, statistically no significant diff erence was observed between two groups (age group >40, <40). With respect to anatomic site of head and neck cancer patients, highest number of HPV+iv was observed in oral cavity 51(63%) followed by pharynx 20(25%), larynx 7(8%) and hypopharynx 3(4%). Disease was more common among illiterate individuals with low socioeconomic status and trend of HPV+ive HNC cases have increaed from year 2011 to 2016. It can be concluded from the data that HPV infection can be the main risk factor for HNC cases in Pakistan. Further research is needed to elucidate the mechanism involved in HPV infection with head and neck carcinogenesis.