Day 2 :
Keynote Forum
Lei Huo
University of Texas, USA
Keynote: MicroRNA expression in advanced breast cancer
Biography:
Lei Huo is a practicing Breast Pathologist in The University of Texas MD Anderson Cancer Center. She is actively involved in clinical and translational research in the field of Breast Cancer. Her research interests include molecular and immunohistochemical markers in tumorigenesis, diagnosis and treatment of breast cancer, among others.
Abstract:
Statement of the Problem: Although early stage breast cancer has a high cure rate with current treatment modalities, advanced breast cancer remains a life threatening disease. There is an urgent need for new therapeutic targets. Inflammatory breast cancer, comprising 1-5% of newly diagnosed breast cancer in the United States, is the most aggressive form of breast cancer, characterized by clinical hallmarks of diffuse erythema and edema and rapid progression from the onset. Recent advances have implicated the role of microRNAs as oncogenes or tumor suppressor genes in tumorigenesis, metastasis and response to treatment in various cancer types including breast cancer. The aim of our ongoing study is to identify microRNA molecules that are regulated in advanced breast cancer, including inflammatory breast cancer. Methodology & Theoretical Orientation: MicroRNA expression profiles of human advanced breast cancer including inflammatory breast cancer were compared to normal breast tissue using a previously validated microRNA microarray assay. The results were subsequently validated by quantitative reverse transcription PCR and in situ hybridization. Findings: There was distinct segregation between tumor and normal breast tissue in microRNA expression profiles. In contrast, between inflammatory breast cancer and non-inflammatory breast cancer, distinct clustering was not readily identified in the microarray analysis. However, several microRNAs were differentially expressed in inflammatory breast cancer. We have validated some molecules by quantitative PCR and in situ hybridization. For example, miR-205 expression was decreased not only in tumor compared with normal breast tissue, but also in inflammatory breast cancer compared with non-inflammatory breast cancer. Lower expression of miR-205 was associated with worse distant metastasis-free survival and overall survival in our cohort. Conclusion & Significance: MicroRNAs may serve as therapeutic targets in advanced breast cancer.
Keynote Forum
Jiangwen Zhang
University of Hong Kong, Hong Kong
Keynote: miRNA Cancer MAP: A web server prioritizing tumor associated miRNA
Biography:
Jiangwen Zhang has completed his graduation from Johns Hopkins University with PhD. He has worked at Harvard University Genome Center as Senior System Biologist for years before joining University of Hong Kong in 2013. His lab has broad interest in genetic and epigenetic regulation in development and diseases. Currently, his lab is focusing on epigenetic regulation of tumorigenesis. His lab employs high throughput ‘omics’ assays and large scale computation to dissect the gene regulatory network and signaling pathways involved in oncogenesis.
Abstract:
Recent studies have revealed the critical role of miRNAs in oncogenesis and great potential of miRNAs serving as diagnostic and prognostic biomarkers. NGS technologies have led to an exponential growth of miRNAs-related data. The mounting body of miRNA NGS data generated requires highly sophisticated tools for data analysis and integration. Currently, there are many tools available for miRNA target identification and miRNA function prediction. An integrated platform incorporating multiple data sources, methods and reported evidences would improve the accuracy and efficiency of the data analysis. Here we present our current work in this direction, miRNACancerMap, a web server inferring miRNA regulatory network in cancers. We collected miRNA-target information from multiple sources such as validated databases and sequenced-based prediction algorithms. Using text mining method, we discovered thousands of miRNA regulations in cancers from PUBMED. We have found 4,879 papers which reported hundreds of microRNAs related to cancer initiation and progression. We have also conducted data mining of miRNA NGS data from many cancer studies, e.g. genome-wide expression profiling of miRNAs and its mRNA targets. Integration of sequence-based miRNA-target interactions and expression-based miRNA-target correlations enable us to distinguish the activated regulations in cancers. And the annotation of cancer-specific evidences and functional analysis facilitate a better interpretation of the miRNA effects on cancer of the miRNA regulatory. Finally, we build up a userfriendly web server, miRNACancerMap, accessible for users to analyze the mRNAs/miRNAs of their interest and their own expression profile. And the results are presented in a comprehensive knowledge map via interactive visualization.
- Cancer Cell Biology l Cancer Biomarkers l Cancer Vaccines
Session Introduction
Tzu Ching Chang
China Medical University, Taiwan
Title: Mesenchymal stem cells in inflammatory microenvironment promotes cancer cell migration and epithelial-mesenchymal transition through osteopontin
Biography:
Tzu-Ching Chang has completed her PhD from Graduate Institute of Life Science, National Defense Medical Center, Taiwan. She did her Master’s degree from Institute of Physiology, National Taiwan University, College of Medicine, Taiwan. Her areas of research include Molecular Cell Biology, Stem Cell Biology and Cancer Cell Biology.
Abstract:
Backgrounds: Mesenchymal stem cells (MSCs) provide potential regenerative effects on chronic kidney disease (CKD) due to their paracrine signaling with cell tropic and anti-inflammation properties. However, in CKD patients combined with renal cell carcinoma (RCC), the roles of MSCs in inflammatory or tumors’ microenvironment are still controversial. Methods & Results: To evaluate the real characteristics of MSCs in inflammatory microenvironment, we first detected the human MSC secretome by cytokine array after MSCs were incubated with or without inflammatory cytokines (IL-1β or TNF-α). The array data showed that, compared to control MSCs, osteopontin (OPN) were significantly increased after MSCs were incubated with inflammatory cytokines. Furthermore, OPN mRNA level and protein secretion were confirmed by quantitative real-time PCR and ELISA respectively. We further collected MSC condition medium (MSC-CM) after inflammatory cytokine treatments to test the effects of MSC-CM as well as OPN alone on RCC migration. MSC-CM and OPN alone facilitated RCC migration by Boydon chamber assay. The epithelial-mesenchymal transition (EMT) of RCC was significantly promoted by MSC-CM and OPN alone since the reduction of E-cadherin and increase of Snail and vimentin. Using OPN-specific antibody to neutralize OPN in MSC-CM would attenuate the RCC migration and EMT that were stimulated by whole MSC-CM. Moreover, the activities of cancer-associated fibroblasts (CAF) were enhanced by treating CAF with MSC-CM and OPN alone. Conclusion & Significance: In this study, we demonstrated that OPN secreted by MSCs would promote cancer cell migration, EMT progression and CAF activation while MSCs were incubated in an inflammatory microenvironment. We hope that this study will provide another consideration of MSC application in kidney regeneration while CKD patients combined with RCC.
Surangkana Sanmai
Khon Kaen University, Thailand
Title: High expression of Pyruvate Dehydrogenase Kinase in Cholangiocarcinoma patients
Biography:
Surangkana Sanmai is currently pursuing a Master’s degree in Medical Sciences at the Faculty of Associated Medical Sciences, Khon Kaen University, Thailand. She is interested in cancer biomarkers and chemotherapeutic target.
Abstract:
In our previous demonstration, we found the differential mitochondrial proteom between CCA and adjacent non-canerous tissues assessed by mass spectrometry. Cancer cells are well-know for their altered metabolic state and the main cause under this phenomenon is mitochondrial metabolic enzyme permutation. So in this study, the glucose metabolism-involved proteins were selected from data and fold-change analysis was performed. The data showed the overexpression of pyruvate dehydrogenase kinase (PDK) isoform 2 and 3 in CCA compared to its adjacent non-cancerous tissues. Especially, PDK3 showed the highest fold-change at 13. PDK is a Ser/Thr kinase that inactivates mitochondrial pyruvate dehydrogenase and plays a keystone of Warburg effect that lead cancer cells survive in hypoxia condition. PDK isoforms consist of PDK1, PDK2, PDK3 and PDK4. All PDK isoforms notably express on specific tissues and there is no report of PDK pattern in CCA yet. In this work, all PDK isoforms immunohistochemistry was performed in CCA and adjacent non-cancerous tissues to confirm mass spectrometric data. The results showed the pattern of PDK isoform expression where PDK1, PDK2 and PDK3 exhibited high expression on CCA tissues at H-score 240±69, 210±30 and 262±28 respectively (maximum H-score=300), while the weak staining was observed in adjacent tissues. In contrast, PDK4 showed no staining in CCA tissues. Taken together, the specification of PDKs pattern in CCA may provide a good tumor marker for CCA. Futhermore, PDK1 and PDK3 may be considered as potential prognostic markers for CCA patients due to their tumor progression involvement.
Doungdean Tummanatsakun
Khon Kaen University, Thailand
Title: Serum apurinic/apyrimidinic endodeoxyribonuclease 1 in cholangiocarcinoma patients
Biography:
Doungdean Tummanatsakun is currently pursuing a Doctor of Philosophy Program in Medical Technology at the Faculty of Associated Medical Sciences, Khon Kaen University, Thailand. She is interested in cancer biomarkers.
Abstract:
Cholangiocarcinoma (CCA) is a cancer from intra- and extra-hepatic biliary epithelial cells. The incidence of CCA in the northeastern part of Thailand is very high, especially Khon Kaen province. Metastasis of tumor provides important prognostic information and can help guide therapy. This study is to discover CCA prognosis marker to distinguish between non-metastatic and metastatic patients from our database in CCA cell lines secretome. Using bioinformatics include SignalP, SecretomeP, TMHMM, Plasma Proteome Database and EMBOSS for selected candidate proteins from three CCA secretome but not in control immortalized cholangiocyte by GeLC-MS/MS. Secretory protein was selected for further validation studies according to criteria including proteins predicted to have signal peptide or non-classical proteins that have no transmembrane helix, identified in serum and selected proteins with maximum of signal peptide cleavage sites. Apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) was selected for further validation analysis. APEX1 has been reported to play a role in metastatic function. Thus, to validate APEX1 protein was differentially expressed in CCA and western blot (WB) for APEX1 was performed for metastatic and non-metastatic group in serum CCA. This revealed that APEX1 intensity ratio was highly expressed in metastatic compared to non-metastatic group (4.73±2.02 for metastatic vs. 1.14±0.81 for non-metastatic group). In conclusion, serum APEX1 might be useful as a prognosis biomarker for treatment in CCA patients.
Daraporn Chua-on
Khon Kaen University, Thailand
Title: High expression of apoptosis-inducing factor, mitochondrion-associated 3 (AIFM3) in human cholangiocarcinoma
Biography:
Daraporn Chua-on is currently pursuing a Doctoral degree in Medical Sciences at the Faculty of Associated Medical Sciences, Khon Kaen University, Thailand. She is interested in cancer biomarkers and chemotherapeutic target.
Abstract:
Diagnostic biomarker of cholangiocarcinoma (CCA) patients is absolutely poor in spite of extensive efforts for the improvment of diagnosis. Mitochondial proteins play key roles in carcinogenesis of various cancer. Therefore, mitochondria are considered as the target organelles for discovering biomarkers of several cancers including CCA. The purpose of this study is to identify potential candidate proteins for diagnosis using mitochondrial proteome analysis for CCA tissues. A shotgun proteomic approch using SDSPAGE coupled with LC-MS/MS was applied to compare the expression of mitochondrial proteins in CCA and the adjacent noncancerous tissues. Using the proteomic analysis for the pooled mitochondrial proteins purified from three each of papillary and nonpapillary types of CCA and their adjacent tissues, 281 proteins were identified as mitochondrial proteins, and 105 of them showed significantly different expression compared with the corresponding counterparts. Among 105 proteins, apoptosis-inducing factor, mitochondrion-associated 3 (AIFM3) was a unique protein commonly over-expressed in both papillary and non-papillary types ot CCA tissues but not in the adjacent non-cancerous tissues. Immunohistochemistry showed expression of AIFM3 staining was significantly higher in CCA than in the corresponding adjacent non-cancerous tissues. Furthermore, high expression of AIFM3 was also significantly expressed in human serum CCA. In conclusion, AIFM3 was aberrantly expressed in both CCA tissues and serum. This finding suggeests that AIFM3 could be a potential biomarker for CCA diagnosis.